1. health and fitness
2. disease
3. cancer

Science and Innovation Week 2009 • Abstracts Book
Healthy City
Sexual and reproductive health
Design and standardization of a multiplex PCR for detection
of bacterial pathogens causing cervicovaginitis
Aguilera AMG1*, González CAM1,3, Méndez TA2*, Juárez ESR3, Castro EG1
1Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional,
2Laboratorio de Bioinformática, Departamento de Bioquímica, Escuela Nacional de Ciencias Biológicas-Instituto
Politécnico Nacional, 3Laboratorio de Pruebas Especiales, Centro Médico Nacional “20 de Noviembre”, Instituto
de Seguridad y Servicios Sociales de los Trabajadores del Estado, México, D.F.
Abstract
Female genital tract’s infections are difficult to recognize through conventional diagnosis methods, mainly when
dealing with difficult managing and recovering microorganisms, this fact makes diagnosis a complex process. The
standardization of a multiplex PCR technique (mPCR), in addition to the conventional diagnosis methodology for
the identification of fastidious microorganisms such as Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma
uraelyticum and Neisseria gonorrhoeae from a single sample may allow carrying out an integral, specific and
sensitive identification. This methodology will offer an opportune diagnosis and therefore decrease the potential
risk of complications associated to misdiagnosed or mistreated infections. The objective of the present report is
to standardize a mPCR that allows the simultaneous identification of the agents mentioned before. If success is
achieved, this PCR may be applied in public health laboratories and contribute as a tool to solve gynecological and
obstetrics problems. Due to the main problem that a mPCR faces up is the optimum selection of primers sequences
that guarantee i. Specificity and efficiency of the amplification and ii. Easily differentiable products of amplification
in an electrophoresis running, in the first part of the present work a careful design and selection of highly efficient
primers to use in the PCR was done. The design was started in silico taking as reference previously described
primers used in simple PCR reactions for each of the microorganisms in study. The selected primers were analyzed,
with an oligonucleotide properties calculator software, to evaluate its physical constants, specificity and hairping/
self dimerization in a mPCR. Finally, modifications of the sequence were done to its optimization so they fulfill all
the desirable characteristics to detect each of the studied microorganisms, including similar melting temperatures
(TM’s) and without the formation of secondary structures. Until now, in vitro standardization of the amplification of
the 4 genes chosen as amplification target have been achieved individually. The simultaneous amplification of N.
gonorrhoeae and M. hominis was achieved as well (duplex-PCR).
This project has got financial support of: ICyT-DF-PICDS08-77 and SIP 20091187.
*Contact
Phone: (55) 5729-6300 ext.62374
e-mail: [email protected] , [email protected]
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Science and Innovation Week 2009 • Abstracts Book
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Sexual and reproductive health
Mammography using laser light pulses
Bruce NC1*, Ortega Martínez R1, Ortíz Rascón E1, Rodríguez Rosales AA1
1Centro de Ciencias Aplicadas y Desarrollo Tecnológico, Universidad Nacional Autónoma de México.
Abstract
One of the most important causes of death in women is breast cancer. It is also known that detection of breast cancer
at an earl stage of its development leads to a very good chance of cure and full recovery of the patient. The accepted
method for screening of the female population for breast cancer is the technique of mammography, or mastography.
This technique involves using X-rays to take an image of the breast to detect the localized changes in density of
the tissue caused by the cancer. To obtain good resolution (details of the fine structure in the image) the breast is
compressed to approximately 5cm width for the exposures, which is very uncomfortable for the patient. Also, there
is a known, and perceived in society, small risk that the exposure to X-rays can actually cause damage to the tissue,
thus increasing the probability of cancer in the patient.
One alternative to the use of X-rays as a screening tool is the use of light. For controlled intensities of light there is
no damage to tissue (high intensities of light are actually used in medicine to destroy tissue: removal of tattoos or
clearing of bloked veins and arteries are two examples), and this is the principal advantage of light-based testing over
X-ray tests. Also, there are many optical sources, components and detectors which have been developed for other
applications but which can be used in medical applications, thus reducing the total cost of optical instrumentation.
However, the main problem with the technique is that light suffers a great deal of scattering inside tissue which
means that a “shadow” image, such as is produced with X-rays cannot be produced with light. To overcome this
difficulty it is possible to use pulses of light produced in special lasers. These lasers can provide pulses as short as
100 femtoseconds (1 femtosecond is 1 thousandth of a millionth of a millionth of a second). These very short pulses
come out of tissue with a much longer duration (of the order of picoseconds which is a thousandth of a millionth of
a second!! or about a million times longer than the input pulse). These pulses can be detected with special detectors
and analyzed. The part of the pulse which emerges from the sample earliest has to be the part of the light which has
traveled in the straightest trajectory from the source to the detector, as this is the shortest path between these two
points. So, this light has information on the fine detail of the image, which is required to detect smaller objects in
the tissue. It may also be possible to perform these measurements without compressing the breast making the test
procedure more comfortable for the patient.
This poster will present the basic ideas behind the technique and the work being performed in the CCADET in this
area.
The authors are grateful to the Instituto de Ciencia y Tecnología del Distrito Federal for supporting this work.
*Contact
Phone: (55) 5622-8602 ext. 1119
e-mail: [email protected]
94
Science and Innovation Week 2009 • Abstracts Book
Healthy City
Sexual and reproductive health
Role of progesterone in women living with HIV/AIDS
and serodiscordant couples
Cabrera ME1,2, Fuentes RLL3, Zamora CJ1, Soto RLE3, Camacho AI1*
1Facultad de Química, Departamento de Biología, Universidad Nacional Autónoma de México, 2Facultad de
Medicina, Departamento de Farmacología, Universidad Nacional Autónoma de México,
3 Departamento de Infectología, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, México, D.F.
Abstract
The Acquired Immunodeficiency Syndrome (AIDS) constitutes the main infectious cause of death in adults worldwide.
Epidemiological data suggest the existence of differences in viral load and CD4+ lymphocytes cell counts related to
gender. Women have more favorable clinical and viro-immunological patterns than men in early infection, although
once established the infection these pattern are reversed. HIV co-receptors (CCR5 and CXCR4) play an important
role in the establishment of infection and disease progression. Caucasian HIV exposed non-infected individuals
(ENI) present a 32 bp deletion in CCR5 that is rare in other racial group, and it is not present in Mexican people.
The explanation for this infection resistance is unknown. Studies in healthy woman have shown that sex hormones
such as progesterone (P) can modulate CCR5/CXCR4 expression through an unknown mechanism. We determined
the effects of P and its intracellular receptor (PR) in the regulation of CCR5 and CXCR4 expression in PBMC of HIV-1
Mexican infected women with typical progression (TP) and ENI. 5x106 PBMC from TP, ENI and seronegative (SN)
controls were cultured in stimulation medium, then it was changed by DMEM without red phenol and fetal bovine
free hormone serum during 24 h. Cells were treated with: Hormone vehicle (cyclodextrin (0.02% in sterile water),
24 h; P (10 or 100 nM), RU486 (P antagonist, 1 µM) 24 h; P (100 nM) + RU486 (1 µM), 24 h. CCR5/CXCR4 content
was determined by Western Blot. Data from protein content densitometry were analysed using Mann-Whitney test.
We found that CCR5 expression was down-regulated after P treatment in SN, TP and ENI, while P significantly
increased the content of the co-receptor CXCR4 in SN and PT. Interestingly, CXCR4 was down-regulated by P in ENI.
RU486 did not block P effects. We suggest that P should play an important role in the acquisition and progression of
infection with HIV-1. Expression of HIV co-receptors in women is regulated by P, up-regulating CXCR4 and downregulating CCR5. Levels of P could influence disease progression in different directions according to co-receptor
use. In addition, P can contribute to the resistance in the acquisition of the HIV by ENI down-regulating both coreceptors.
This Work was supported by Instituto de Ciencia y Tecnología del Distrito Federal, México, grant PICDS08-35.
*Contact
Phone: (55) 5622-3869
e-mail: [email protected]
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Science and Innovation Week 2009 • Abstracts Book
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Sexual and reproductive health
NTS-polyplex as a potential tool in gene therapy for human
breast cancer: evidences in vitro
Castillo Rodríguez RA1, Arango Rodríguez ML1, Escobedo L1, Rubio Zapata HA2, Rembao Bojorquez JD3,
Sánchez García A3, Dupouy S4, Forgez P4, Martínez Fong D1*
1Departamento de Fisiología, Biofísica y Neurociencias, Centro de Investigación y de Estudios AvanzadosInstituto Politécnico Nacional, 2Escuela de Medicina, Universidad Autónoma de Yucatán, 3Departamento de
Neuropatología, Instituto Nacional de Neurología y Neurocirugía, México D.F, 4INSERM UNRS 938, Paris, France.
Abstract
The expression of the high affinity neurotensin receptor (NTSR1) is induced in the invasive ductal breast
adenocarcinoma, which is the cancer subtype with the highest incidence and mortality rate in the whole world. Our
group has developed a new antitumoral therapy for NTSR1-expressing cells using the gene transfer system known as
NTS-polyplex (patente mexicana # 264932). The main objective of this work was to determine in vitro the ability of
MCF7 and MDA-MB-231 cell lines, from ductal breast adenocarcinoma, to be transfected by the NTS-polyplex, as
a first approach of antitumoral therapy.
We used internalization and expression assays with reporter genes (pEGFP-N1 and pORF-Luc) in combination with
blockade studies with neurotensin (1 µM), SR48692 (0.5 µM) or sucrose (0.45 M). For functional studies, we used
the suicide system pORF-HSVTK-ganciclovir (GCV). Cytotoxicity was evaluated using MTT assay and annexin-V.
RT-PCR and immunofluorescence studies confirmed the presence of NTSR1 in those cell lines, which were able to
specifically internalize and express reporter genes. The MDA-MB-231 cell line was more efficiently transfected than
MCF7 cells were, but the viability of MDA-MB-231 cells was decreased (60%) by the transfection procedure. In
these cells, the transfection of pORF-HSVTK plasmid decreased cell viability by 50%, in an independent manner of
the activation with GCV. The blockade assays strongly suggest that the cytotoxic effect was caused by the NSTR1mediated endocytosis of NTS-polyplex. In conclusion, MDA-MB-231 cells were more susceptible to NTS-polyplex
transfection, which was cytotoxic for this cell line. This particular property could represent an additional advantage for
a more effective antitumoral therapy in models of breast cancer. This work was supported by the Instituto de Ciencia
y Tecnología del Distrito Federal (grant ICyTDF-DMF) and partially by SEP-Conacyt-ECOS (Grant M07-S01).
*Contact
Phone: (55) 5747-3800 ext. 3959
e-mail: [email protected]
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Sexual and reproductive health
HIV epidemiological models
Fonseca N1, Martínez CLP2, Zárate S1*
1Posgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de México,
2 Facultad de Química, Universidad Nacional Autónoma de México. México, D.F.
Abstract
Introduction: The spread of HIV in a population depends on a variety of biological and social factors. The genetic
makeup of the host and the fitness of the virus transmitted to a particular host are likely to impact the likelihood of
transmission. Also, since the spread of HIV infection depends largely on the social dynamics of the population, it
becomes necessary to introduce structure in the way individuals interact with one another. Furthermore, different
subgroups within the population show differences in their risks of infection.
Methodology: Agent-based models were used to recreate the dynamics of different epidemics. We varied a set of
characteristics among the population, both of biological nature, such as susceptibility to the infection, and of social
nature, like the tendency to engage in intercourse. After multiple runs of the simulations were obtained, the results
were analyzed by using generalized linear models to determine the effect of the variables in the spread of the
infection.
Results: We found that all the behavioral variables contributed significantly to the fit of the model (p < 0.001),
with the tendency to stay in long term relationships being the factor that contributed the most to change the rate of
spread of infection. On the genetic level, the distribution of susceptibility of the individuals was the most important
contributor to halt the epidemic, as expected. The introduction of structure in the population was crucial to identify
which intervention strategies are more likely to become successful.
Conclusions: The use of individual-based models allowed us to mimic the complex dynamics of the epidemic and
the social network underlying it, and to find the components with the larger impact on the spread. Also, it was
possible to include a complex social network in the model, which is necessary to develop reliable models of HIV
spread.
*Contact
Phone: (55) 5488-6661 ext. 15160
e-mail: [email protected]
97
Science and Innovation Week 2009 • Abstracts Book
Healthy City
Sexual and reproductive health
Photoacoustic tomography: on the development and application
of photo-thermal sensor arrays for breast cancer imagenology
García Segundo C1*, Quispe Sciccha RM1, Garcés Madrigal A1, García Valenzuela A1,
Guadarrama Santana A1, Sánchez Pérez CA1
1Centro de Ciencias Aplicadas y Desarrollo Tecnológico (CCADET), Universidad Nacional Autónoma de México,
México, D.F.
Abstract
Worldwide, Breast Cancer is ranked as the fifth cause of death among women. Yearly the international community
dedicates large amount of efforts and resources for achieving its medically reliable diagnosis, however the early
diagnosis is a problem yet to be resolved. The effort of reducing the deadly cost applies mainly to largest urban areas,
like Mexico City. The general statistics exhibits that apart the genetic predisposition to acquire cancer, the urban-life
habits and the excessive population concentration (which also reduces the access to medical services) are also rather
important factors. Being Mexico City among the world’s top five biggest urban areas, it is rather clear that this city
has serious requirements of attention and concern on this type of health issues.
The present project is focused on developing science and technology aid for the early diagnosis of breast cancer, by
use of the non-invasive laser induced photoacoustic (PA) method. The premises for the current development are: 1)
we want to avoid the trend of the mastography-like diagnosis methods which are quite invasive, traumatic and not
always conclusive; 2) it is well established that death rates are substantially reduced, with possibilities to get back
a normal life, if early diagnosis is provided; 3) it is cheaper for the patient to cover the cost of early diagnosis rather
than the costly: late diagnosis and therapy.
The core of the photoacoustics phenomenon in materials like the human tissue is to generate acoustic-like waves
(PA-waves) out of the absorption of pulsed laser energy at near-infrared (NIR) wavelengths. The human tissue is near
transparent for illumination at NIR (within 800 nm and to 2000 nm, approximately), however the blood’s haemoglobin
and oxi-haemoglobin absorb energy within this range. The bottom-line is that breast vascular concentrations, thus
haemoglobin and oxi-haemoglobin excess, are the definitive mark for the presence of carcinomas. Our development
around PA-Tomography consists in developing sensor-arrays and associated electronics to recover laser induced PAwaves, out of lab samples with breast-like properties. The recovery of these signals through matrix-like sensor-arrays,
brings up the fundamental conditions to reconstruct and locate the absorption centres (breast tumours) in image
format.
*Contact
Phone: (55) 5622-8602 ext. 1122
e-mail: [email protected]
98
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Sexual and reproductive health
Development of low-cost DNA microarrays for the identification
of gene expression profiles of tumor markers in cervical cancer
Hernández Mejía A1, Marcial Román I1, Álvarez Ríos E1, Herrera JL1, Reyes Ibarra AP1,
Ocadiz Delgado R1, Gariglio Vidal P1*
1Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados-Instituto
Politécnico Nacional, México, D.F.
Abstract
Cancer has become one of the main causes of mortality in Mexico, with roughly 12% of deaths per year; this disease
constitutes an important socioeconomic and health problem, because affects mainly people in productive age and its
treatment is expensive. In Mexico City approximately 9,800 persons die each year for some type of cancer. Cervical
cancer (CC) is one of the main causes of mortality by cancer of the Mexican women. In Mexico City around of
550 women die each year for this disease justifying a detailed study. High-risk human papillomaviruses (HR-HPV)
mainly types 16 and 18 are considered risk factors in cervical cancer. However, the infection with these viruses is
not sufficient to induce tumors and a lot of efforts are concentrated on the study of cofactors involved, as well as
the identification of molecular markers useful for early diagnosis of CC. Recently, modern techniques focused to
molecular diagnostic have been developed, which allow the simultaneous expression analysis of thousands of genes
in a single assay. They employ cDNA or DNA, corresponding to the sequence of the genes to analyze. These arrays
are situated in small areas on a chemically activated solid surface, allowing bi-dimensional arrangements composed
of hundreds or thousands of these sequences. This technology and the information derived from the sequencing of
the human genome, is currently applied in basic research and in areas related to biomedical diagnosis, prognosis and
pharmacogenomics. In this project we have developed low-cost microarrays using DNA obtained from clones with
specific tumor markers previously described in the literature, such as p16, c-myc, n-myc, mmp10, cldn1 and rho-c,
junb, birc5, mdm2, rb1, pcna, ub2e, cyca, cycb, cyce, bcl2, p21, klk10, il6. These markers were bound in both lowcost and commercial slides. In the stage of standardization, we have obtained the expression profiles of HR-HPVpositive cell lines derived from CC, as a validation model employed during the production process. We have already
obtained the conditions for isolation, purification and amplification of each tumor marker. We have also optimized
printing protocols using “A+ Nexterion Slides” (Schott; USA), as well as the labeling and hybridization of targets. We
determined the expression profile of tumor markers mentioned previously, in the following cell lines derived from
CC: SiHa (HPV16+), HeLa (HPV18+) and ViBo (HPV-). The signal showed high specificity for each tumor marker,
and interestingly there was correlation with the expression levels described previously for each cell line. Expression
levels were corroborated (mmp10 and rho-c) using real time RT-PCR. Home-made slides (CICATA-IPN) satisfy the
necessary specifications for the construction and use of microarrays. In conclusion, we have developed low-cost
DNA microarrays for the identification of gene expression profiles of tumor markers in cervical cancer.
*Contact
Phone: (55) 5747-3800 ext. 3337
e-mail: [email protected]
99
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Voltage-gated sodium channels in cervical cancer: potential
use as biomedical diagnosis
Hernández Plata E1, Díaz Velázquez CE1, Ortiz Arce CS1, Rivera M1, Gómora J1*
1Departamento de Biofísica, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, México,
D.F.
Abstract
The cervical cancer (CaC) is the third cause of death among women in the world. This anomalous cellular proliferation
of the cervical endothelium is associated with infection by oncogenic types of human papillomaviruses (HPVs).
A big challenge that still remains in CaC is the lack of new tools for an accurate detection of this carcinoma. In
this regard, several recent reports have attributed different roles to ion channels in the regulation of signal during
cell proliferation, invasiveness and metastasis. For instance, it has been demonstrated that voltage-gated sodium
channels (VGSC or NaVs) are functionally expressed in highly metastatic cancer cells derived from non-excitable
epithelium and its activity has been associated to cell motility and invasiveness in breast and prostate cancer. In both
cases, the VGSC have been proposed as new potential molecular markers for such types of cancer. During the last 5
years we have been studying the role of VGSC in the biology of CaC, having demonstrated the functional expression
of this kind of channels in primary cultures derived from human CaC biopsies. In addition, we also reported the
expression of mRNA for several VGSC -subunits. Our most recent Real Time PCR experiments indicated that two
VGSC -subunits are particularly over expressed in CaC respect to normal cervix biopsies: NaV1.6 and NaV1.7b.
Electrophysiological evidence suggests also that these -subunits actually contributed to the global sodium current
recorded from CaC primary cultures cells. Based on these findings, we suggest that VGSC might play an important
role in several cellular functions related to the progression of CaC carcinoma (i.e. proliferation, invasiveness, etc.).
Finally, due to the selective over expression of NaV1.6 and NaV1.7b, these VGSC a-subunits might be considered as
potential biomedical diagnosis of CaC.
*Contact
Phone: (55) 5622-5752
e-mail: [email protected]
100
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Brigdering epidemiology and demography: considering
a modern approach to breast cancer epidemiology
Herrera Becerra AA1*, Rosendo Robles A1, Padrón Godínez A1, Prieto Meléndez R1
1Grupo de Modelado y Simulación de Procesos, Centro de Ciencias Aplicadas y Desarrollo Tecnológico,
Universidad Nacional Autónoma de México. México, D.F.
Abstract
Epidemiology is defined as the study of the distribution and determinants of health-related states or events in specific
populations, and the application of this study to the control of health problems. As such, epidemiology is considered
by many thinkers and practitioners as an independent branch of Public Health; this, in turn, is defined as an organized
community effort to prevent disease and promote health. Epidemiology is a relatively new scientific discipline; its
origins are commonly set at the middle of the XIX century in England; in fact, epidemiology and demography were
born in the same period and, until the turn of that century, they were considered as interwoven activities. In spite
of its short history, the discipline has earned great recognition for its enormous contributions to the understanding
of the determinants of health-related states in populations. Nevertheless, epidemiology has also faced some serious
drawbacks and criticisms. In particular, at the turn of the XX century, some renowned thinkers in the field have been
alerting that the discipline is dangerously losing its original population and socioeconomic perspective, in favor of a
more individual perspective. They claim that there is a current lack of interest in studying and considering population
factors as cause of disease; that is, epidemiology has diverged from demography. These thinkers have reasons to
postulate that the dominant epidemiological paradigm in the XXI century will be one that reverses its separation
from demography.
In this context, we have been reviewing the development of epidemiology in the past century, analyzing the transition
from the contagious disease (single factor) paradigm to the chronic disease (multiple factors) paradigm, and paying
attention to the shift in methods of study and in conceptions of disease or health state. We want to understand
the methodological, conceptual and political reasons that eventually led to the separation of epidemiology from
demography. Initially, we are following the proposals of thinkers as M. Susser, E. Susser, and D. Lilienfeld. The
ultimate objective of our study is to establish a modern reference framework suitable for developing epidemiological
studies of cancer, specifically breast cancer. We present our initial findings and discuss the immediate direction we
think our investigation must take.
*Contact
Phone: (55) 5622-8602 ext. 1188
e-mail: [email protected]
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Computer-assisted system for early breast cancer detection
Hevia Montiel N1, Lira Berra E1, Flores Mijangos M1, Arámbula Cosío F1*
1Laboratorio de Análisis de Imágenes y Visualización, Centro de Ciencias Aplicadas y Desarrollo Tecnológico,
Universidad Nacional Autónoma de México, México, D.F.
Abstract
Breast cancer is one of the leading death causes by malignant tumor for women in Mexico, in agreement with
information reported by the “Instituto Nacional de Estadística, Geografía e Informática” (INEGI), along with the
cervical-uterine cancer with 13,3% and 13,5% of deaths, respectively. Breast cancer is the main cause of mortality
in the group of women between 30 and 64 years of age, who are in their more reproductive stage. Unfortunately,
only 5 to 10% of the diagnosed cases are in the early clinical phase (0 and 1 BIRADS), but this percentage data is a
underestimation of the reality, since the early detection programs of breast cancer, as well as the number of specialists,
and the technical equipment for diagnosis (X-ray mammographs) are insufficient to cover the population at risk.
Digital image processing and computer analysis of medical images can provide valuable quantitative information (the
morphometric measurements, density distribution functions, attributes of texture, etc.) and qualitative information
(enhancement and restoration, local and highly specific transformations in zones with determined morphologic
properties). Together these techniques can effectively assist the expert radiologist in the detection of breast cancer in
x-rays mammography.
The objective of this research project is to develop a computer-assisted system for the early detection of breast cancer
using digital image processing and analysis techniques. The system will provide automatic functions for the detection
of tumors and micro-calcifications in digital mammography images. The system will highlight suspicious regions in
the mammography to the expert radiologist, who will perform the diagnostic. In Fig. 1 are shown the results of the
application of a contrast enhancement filter (IRIS filter) to a digital mammography image.
(a)
(b)
(c)
Figure 1. Example of digital processing of a mammography image: a) original image, b) and c) IRIS filtered images
at different filter sizes
In case that the diagnosis on mammography images indicates that it is necessary to perform a tissue biopsy. Our
computer-assisted system will be able to assist the radiologist during the realization of a breast biopsy guided with
ultrasound imaging. An optical tracking system will be used for the acquisition of the ultrasound images and for the
update, in real time, of the needle position with respect to the tumor. Computer graphics models of the tumor and
the needle, updated in real time, will be used to guide the radiologist during the procedure. The 3D graphics model
of the tumor will be constructed using the contours of the tumor segmented on a set of tracked ultrasound images.
Figure 2 shows the breast biopsy computer-assisted system.
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The same computer-assisted system will be suitable for training, of a radiology resident, in the procedure of breast
biopsy, since it will provide the three-dimensional position of the tumor, with this system a student should learn to
orientate the biopsy needle using ultrasound images with shorter learning times.
Figure 2. Computer assisted breast biopsy system
*Contact
Phone: (55) 5622- 8602 ext. 1148
e-mail: [email protected]
103
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Homocisteine and asymmetric dimetil arginine as early nutritional
markers for preeclampsia
López Alarcón M1*, Hinojosa Cruz JC1, Montalvo Velarde I1
1Unidad de Investigación Médica en Nutrición, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro
Social, México, D.F.
Abstract
Serum concentration of homocysteine (Hcy) and asymmetric dimetil arginine (ADMA) are usually increased in
clinical conditions that are characterized by vascular damage. Concentrations of these molecules are decreased
during normal pregnancy, but have been found elevated in patients with preeclampsia (PE). However, it has not
been established if such alteration occurs before the onset of the clinical syndrome, which is important because if
so, routine measurements throughout pregnancy would help to identify those women at risk. Hcy and ADMA are
closely related to folic acid (FA) and vitamins B6 (VB6) and B12 (VB12), but it is also unknown whether alterations
of Hcy and ADMA during pregnancy are associated with deficiencies of these vitamins.
Aim: To evaluate the utility of Hcy and ADMA determinations throughout pregnancy to identify women at risk to
develop PE, and to assess if alterations in these molecules are related to the nutritional status of FA, VB6 and VB12.
Methods: A longitudinal study with a case control design nested in a cohort was proposed. A sample of 750 women
coursing with a normal pregnancy prior to week 20 of gestation will be included and followed until parturition. At
selection, medical and dietary histories will be obtained as well as a peripheral blood sample for Hcy, ADMA, VB6
(HPLC), FA and VB12 (RIA) determinations; this procedure will be repeated monthly until parturition. Comparison
will be conducted between women with and without PE.
Results: At present, 154 women have been selected. Seventy-four completed the follow-up, four developed PE and
17 delivered prematurely (PT). Hcy and vitamins have been determined in PE, PT, and in 19 with normal pregnancy
(NP). Serum Hcy decreased throughout pregnancy and recovered basal levels at postpartum; no differences between
groups are present. VB6 and VB12 decreased during pregnancy and recovered basal values at postpartum similarly
in all three groups. In contrast, FA diminished throughout pregnancy and continued declining at postpartum. No
measurements of ADMA have been conducted to date.
*Contact
Phone: (55) 5627-6900 ext.21944
e-mail: [email protected] , [email protected]
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Maternal education, socio-economic level, folic acid intake and birth
defects: a clinical epidemiologic approach in hospitals of the Secretary of
Health of Mexico City
Mutchinick OM1*, Arteaga J1, Luna L1
1Departamento de Genética, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán y Secretaria de
Salud del Distrito Federal. México, D.F.
Abstract
Background: Birth defects, in particular congenital malformations (CM) are an important problem of public health
all over the world. Approximately, the prevalence in livebirths is 2.5%, figure that increase to ≈ 12% in stillbirths.
In other words 1/40 and 1/8 live or dead newborns has a CM. Although with some regional variation and some
particular pattern of prevalence of some CM, the same occur in the Mexican population.
Objectives: To identify possible genetic, reproductive and environmental risk factors associated to the occurrence of
CM in general and to some CM in particular. Material and methods: The sample analyzed includes 140 malformed
and 140 controls of the same sex and hospital from a total of 13,423 births born in seven hospitals of the SHDF. A
special designed file for the study that includes a number of variables respect the child, family, genetic, reproductive
and environmental hazards was obtained for each case and control from the obstetric history and maternal direct
inquire. All babies were examined by a qualified pediatrician who made diagnosis of the defect, selected the control
and collected the data. A database was designed for the coded information obtained and analyzed by members
of the Department of Genetics responsible of the study. Results: Maternal education showed a higher proportion
of lower schooling of mothers of cases than control ones, although maternal age was quite similar. However, a
high prevalence of young mothers (<20 years of age) was observed in the groups of mothers of cases and controls,
24.3 and 23.6% respectively. The proportion of families with a lowest socio-economic level (SEL) was significantly
higher in cases families (46.5%) than controls (32.2%). Folic acid (FA) diet supplementation was less frequent
made by mothers of cases (74.1%) than by mothers of controls (83.0%). Although the proportion of mother being
supplemented was high, the correct periconceptional recommended use was very low (9.52%). Other interesting
findings were: a higher proportion of males affected, due to significantly more males with congenital heart defects
and gastrointestinal atresias, a lower weight and length in malformed newborns than controls and a higher frequency
of premature membrane rupture in case pregnancies. Discussion: Present results confirm previous observations that
low maternal education, very young mothers and low socio-economic level are associated risk factors to CM. Also
make evident, that although FA supplementation is of public domain, the prescription and public knowledge of
the correct daily intake of 400mcg is not observed by the great majority of pregnant women, in part due to the low
prevalence of planned pregnancies. The above shows the need of massive health education programs concerning
risk factors to prevent the occurrence of congenital malformations.
*Contact
Phone: (55) 5487-0900 ext. 2514
e-mail: [email protected]
106
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Sexual and reproductive health
Reliable operation and safe transportation of a mastography
instrument on a mobile unit
Nava Sandoval R*1, Ruiz Botello GA1, Orduña Bustamante F1, Pérez Ruiz JS1, Pérez López A1
1Centro de Ciencias Aplicadas y Desarrollo Tecnológico (CCADET), Universidad Nacional Autónoma de México.
México, D.F.
Abstract
This Project belongs to the Healthy City Program launched in 2008 and managed by the Mexico City Government
(GDF) throughout its Science and Technology Institute (ICyTDF), and is related with sexual and reproductive health,
among other subjects.
Mammal cancer prevention in México City is an important activity carried out by GDF with the collaboration of
several NGO. Particularly, Inmujeres DF, an arm of GDF in charged to attend the problematic of women in Mexico
City, works closely with FUCAM, an NGO which is devoted to prevent and attend mammal cancer diseases. In
accordance with Inmujeres DF, FUCAM manages several mobile units. Each one of these mobile units carries on
2 mastography instruments, and travels through the City, covering the different residential areas on a programmed
schedule. By the many times very adverse travel and “in situ” operation conditions, the mastography instruments
could be exposed to shock and mechanical vibrations, as well as temperature and humidity changes, power
supply fluctuation and electromagnetic noise presence, among other influence factors. These effects could affect
the mammography instruments operation conditions and resulting mammograms, and eventually, to impose the
necessity to take out of service the mobile unit, impacting adversely Inmujeres DF and FUCAM programs.
Attending an specific request coming from the GDF Secretary of Health, CCADET-UNAM, with the support of
ICyTDF, -on the basis of its wide experience to solve problems on mechanical vibrations in instrumentation and
design and construction of prototypes for public and private sectors-, proposed this project in order to provide a
solution which could help to diminish the adverse effects involved on the mobile units efficacy and efficiency. To do
this, CCADET-UNAM has planned to accomplish the following objective:
• To develop a prototype of a system for reliable operation and safe transportation of a mastography instrument on a
mobile unit, to assure quality results for patients and accomplishing Inmujeres DF and FUCAM’s specifications.
The impact expected from this project reaches some sectors of interest, namely:
• Social sector: assuring opportune and quality service for the impacted population.
• Educative sector: by means of students training around the Project.
• Research and development sector: investigating, developing and applying methods of measurement of
influence factors and data analysis; designing and manufacturing prototypes; as well as publishing scientific
and technical papers and reports.
• Intellectual property sector: by means of patent registration and technology transfer, if applicable.
*Contact
Phone: (55) 5622-8602 ext. 1145
e-mail: [email protected]
107
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Sexual and reproductive health
Photodynamic therapy effect using ALA and the inhibitor of histone
deacetylases NaB in cervical and breast cancer cells
Ramón Gallegos E1*, Castillo Millán J1, Vargas Ramírez AL1, Arenas Huertero FJ1, Orea Cruz A2,
Uribe Hernández R1
1Laboratorio de Citopatología Ambiental, Escuela Nacional de Ciencias Biológicas-Instituto Politécnico Nacional,
2Departamento de Física, Centro de Investigación y de Estudios Avanzados -Instituto Politécnico Nacional,
México, D.F.
Abstract
The breast cancer (BCa) and the cervical cancer (CCa) are the first and second in frequency in Mexico, respectively,
thus, they are considered as a health public problem (INEGI, 2008). The therapies used up to now are partially
effective (70%) and the side effects that present, at times, lead to death. Therefore it is necessary to search for new
therapies with a high degree of specificity with minimal side effects or without them. In addition, most of the patients
with CCa are poor and find it difficult to maintain an expensive treatment such as those uses now. The precursors of
each therapy, propose in this work, are inexpensive and available in its acquisition, then the combination of therapies
is adapted to the economic reality of the country. In this work it were studied two highly specific therapies for the
cancerous cells, the photodynamic therapy (PDT) and the chromatin modification, that had not been used combined
in the CCa and the BCa. The aim of this work was to increment the efficiency of the PDT, using δ-aminolevulinic acid
(ALA) and the inhibitor of histones deacetylases, sodium butyrate (NaB), in CCa and BCa cells. It were use the CCa
cell lines: HeLa, SiHa and C33; and the BCa cell lines: MDA-MB-425 and MCF-7. In both cases the HaCat cell line
was used as control. It were determined the appropriate times and concentrations of NaB to relax the chromatin and
ALA to determine the concentration that induced the largest accumulation of PPIX as well as the energy densities
suitable for the irradiation. The conditions used were: 40 µg/ mL ALA for the MDA cells and 80 µg/ mL ALA for MCF7, HeLa, SiHa, HaCat and C33 cells, the NaB was used at 4 mM. CCa cells were irradiated at 64.3 J/cm2, twice,
and the BCa were irradiated, once, at 120 J/cm2. With regard to the photodynamic therapy removal efficiency of
the CCa cells it was found that the PDT eliminated alone: 96.3% of HeLa cells, SiHa of 95% and 85% of C33, when
the PDT was combined with the chromatin modification therapy that involves the use of NaB, the cells deaths were
similar. The photodynamic therapy removal efficiency of BCa cells that was quantified by the plasmatic membrane
damage (neutral red method) and the alteration in metabolism at mitochondrial level (Alamar blue method) was
of 97% in MDA-MB-231cells (hormone-independent) and 30% in MCF-7 (hormone-dependant) by metabolism
damage. However, when both therapies were combined the damage was more evident in the plasmatic membrane.
An elimination of 92% of MB-231 cells and 80% of MCF-7 cells was observed. Apparently the NaB only improves
the response of the PDT to eliminate the cancerous hormone-dependent cells of breast cancer. In the in vivo model
the CCa tumor the PDT can provoke death cell by apoptosis of the 90% of the implanted cells. So, we can conclude
that the PDT, in in vitro conditions, showed a higher efficiency in the elimination of CCa and BCa cells than the
therapies used nowadays in the patients with this type of cancer.
*Contact
Phone: (55) 5729-6300 ext. 62466
e-mail: [email protected]
108
Science and Innovation Week 2009 • Abstracts Book
Healthy City
Sexual and reproductive health
Identification of cysteine proteinases as potential biomarkers for diagnosis
of trichomonosis by an immunoproteomic approach, using the active
degradome of Trichomonas vaginalis and patient sera
Ramón Luing LA1, Rendón Gandarilla FJ1, Cárdenas Guerra RE2, Rodríguez Cabrera N2, Ortega López J2, Ávila
González L1, Ángel Ortiz C1, García Mendiola R1, Herrera Sánchez CN3, Mendoza García M3, Arriaga Alba M4,
Puente Rivera Jonathan1, Flores Robles D5, Alarcón Romero L del C6, Ortega López J7, Arroyo Verástegui R1*
1Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados-Instituto
Politécnico Nacional, 2Departamento de Biotecnología y Bioingeniería, Cinvestav-Instituto Politécnico Nacional,
3Laboratorio de Bacteriología del Laboratorio Central, Hospital General de México, 4Laboratorio de Investigación
Microbiológica del Hospital Juárez de México, 5Unidad de Investigación Especializada en Microbiología,
6Laboratorio de Citopatología de la Facultad de Ciencias Químico Biológicas de la Universidad Autónoma de
Guerrero, 7Hospital de Primer Contacto del Grullo, Jalisco.
Abstract
Trichomonas vaginalis is a human pathogen responsible of trichomonosis, one of the most common non-viral sexually transmitted
infections, which is associated with serious health complications for women and men. T. vaginalis has many cysteine proteinases (CPs);
some of them are involved in trichomonal pathogenesis. Their relevance during infection has been established: a) antibodies against
some CPs were detected in trichomonosis patient sera; b) some CPs have been found in vaginal secretions of patients with active
trichomonosis. Although, trichomonosis can be diagnosed by different methods, most of them have serious limitation or are expensive;
the main problem of the trichomonosis diagnosis is that at least 50% of the infected population is asymptomatic. Therefore, a challenge
is to design an alternative, specific, highly sensitive, and inexpensive diagnostic method, by selecting antigenic proteinases that could be
used as biomarkers to detect the presence of T. vaginalis antibodies in blood samples of asymptomatic patients or patients with vaginitis
or urethritis. Thus, the goal of this study was to identify the potential antigenic proteinases of T. vaginalis and validate them as biomarkers
for serological detection of patients with active trichomonosis. In this study, 1574 enrolled patients attending Hospitals from Mexico
City, Jalisco, and Guerrero who accepted to participate in this study by a signed consent donated biological samples (blood and vaginal
secretions) following the hospital bioethics regulations. These patients were diagnosed with vaginitis by clinical and microscopical
analysis of their vaginal secretions. Trichomonosis was confirmed in 61 of the 1574 patients by in vitro culture of their vaginal specimens
in the InPouch TV test. To detect potential biomarkers of trichomonosis, serum from four culture-positive patients were used to perform
immunoproteomics analyses, by combining the two dimensional-gel electrophoresis (2-DE), mass spectrometry analysis, and Western
blotting (WB) techniques. The major reactive spots detected by T. vaginalis positive patient sera corresponded to proteinases TvCP2,
TvCP4, TvCP4-like, TvCPT, and TvLEGU-1; four papain-like and one legumain-like CPs, respectively. The genes of three of them, TvCP4,
TvCPT, and TvLEGU-1, were cloned and expressed in Escherichia coli, and used as antigens to validate these CPs as biomarkers. For
their validation, Western blot analysis was performed with the three recombinant CPs as antigens and ten patient sera (8 Tv-positive and
2 Tv-negative). Our results indicate that these CPs are indeed biomarkers for the diagnosis of trichomonosis. Thus, these recombinant
CPs could help to detect active trichomonal infection in asymptomatic patients or patients with vaginitis or urethritis by an ELISA-based
assay that can be easily applied in almost any clinical laboratory. In order to develop an ELISA-based diagnostic kit prototype, all the
samples that we have now and more that will be obtained need to be tested to determine the best combination of recombinant CPs for
specificity and sensitivity, as compared with the in vitro culture assay, the gold standard. In addition, throughout this study, we have
provided trichomonosis diagnostic information to the enrolled hospitals, as a public service. This information has been contributing to
improve the treatment of the patients with trichomonosis.
This work was supported by grants 68949 and 58611 (to R.A.) from Consejo Nacional de Ciencia y Tecnología (Conacyt) Mexico and
a grant from Instituto de Ciencia y Tecnología del Distrito Federal (ICyTDF) (to R.A.).
*Contact
Phone: (55) 5747-3800 ext. 5667
e-mail: [email protected]
109
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Sexual and reproductive health
Internal Guideline II “Detection and integral following of mammal cancer”
Rivas Pastor ES1, Vélez Andrade C1, Rivera Ríos MA1*
1Instituto de Investigaciones Económicas, Universidad Nacional Autónoma de México. México, D.F.
Abstract
It’s a guideline which proposes a model (termed Lose Opportunities in Mammal Cancer) to adoption, implementation
and maintenance of the national rules to prevention, diagnose, treatment, control, epidemiologic watching in
mammal cancer. It helps to the health team, patients and susceptible women. Includes specific data and element
needed to meet individual requirements; show all knowledge about mammal cancer to obtain the care, clinical
and epidemiological services on precise time, through the primary, secondary and tertiary preventive levels, by the
way integral health attention to an with de patient, family, social groups, and community, together, improving their
quality of life.
It ofers18 tables
The ended goal is decrease mortality rates of mammal cancer in Mexico City.
All data should be digitalized using System Castor, and be able for all health institutions.
*Contact
Phone: (55) 5603-3600
e-mail: [email protected]
110
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Sexual and reproductive health
Internal Guideline III “ Lose health opportunities in basic programs,
by preventive levels”
Rivas Pastor ES1, Vélez Andrade C1, Rivera Ríos MA1*
1Instituto de Investigaciones Económicas, Universidad Nacional Autónoma de México. México, D.F.
Abstract
This guideline is intended as a support for the health team and community, and also helps as a technical assistance
tool to national and local health units and healthy basic programs, making possible patient’s transferred and antitransferred. They be able to make hard relations between private and public health agencies, social security, helping
social organizations, patients, their family and community into basic programs: AIRD, ADD, Diabetes, AH, Pregnancy,
Healthy Children Control, Elderly People.
All data should be digitalized using System Castor and be able for all health institutions.
*Contact
Phone: ( 55) 5603-3600
e-mail: [email protected]
111
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Sexual and reproductive health
Proteomic analysis of the interaction of Trichomonas vaginalis
with prostatic cells
Vázquez Carillo LI1, Quintas Granados LI1, Arroyo Verástegui R2, Castañón Arreola M1, Álvarez Sánchez ME1*
1Posgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de México,
2Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios AvanzadosInstituto Politécnico Nacional, México, D. F.
Abstract
Introduction: Trichomonas vaginalis is the causative agent of trichomonosis, a common infection worldwide. The
trichomonosis in women cause vaginitis, cervicitis and urethritis. In men, this infection is frequent asymptomatic
although sometimes it causes urethritis and chronic prostatitis. The iron is an essential element in T. vaginalis and is
a component of vaginal microenvironment. This cation modulates the expression of crucial metabolic enzymes and
several pathogenic properties in this parasite. Recently, the proteomic allows a description as well as a characterization
of cell protein expression in presence or absence of iron. A reference map of soluble proteins in the pH range 4–7
used a fresh clinical isolate grown in presence or absence of iron, this map was created by using a combination
of 2DE protocols and tandem mass spectrometry. However, our understanding of the interaction of T. vaginalis
with prostatic cells remains uncertain. The Zn2+ is an important element of the prostatic fluid and plays a crucial
role in the immunology of the infections and pathology of the prostate. The normal high Zn2+ concentrations in
human prostatic secretions (4.5 to 7 mM) appear to be important in preventing the men trichomonosis. Nevertheless,
Zn2+ concentration is lower than 1.6 mM, it is not trichomonicidal and these levels are found in patients with
chronic prostatitis and prostate cancer. We propose that a proteomic investigation focused on protein expression of
T. vaginalis in presence or absence of Zn2+ might contribute to understand the role played by this cation.
Objective: of the present study was to investigate the Zn2+ dependent changes in the proteome of T. vaginalis when
the parasites interact with the cells from prostate of human cancer, called DU145 that had been used as a model for
studying the interaction of T. vaginalis.
Results: Preliminary, the analysis by 2DE gels showed difference in number of spots in Zn2+concentrations, the
proteins were distributed into pH ranges 4-7 with a molecular mass range between 20 and 60 kDa. The 2DE protein
spot profiles were highly reproducible in terms of the total number of the protein spots and their relative positions and
intensities. The images of three representative 2DE gels obtained from three independent experiments were analyzed
by the pDQuest software; one comparative analysis of these gels was used for developing proteomic expression map
of T. vaginalis. Also, we investigated how the levels of Zn2+ affect the cytotoxicity and the proteolytic activity of
CP65 in this parasite; we found that the levels and the proteolytic activity were reduced in the concentration of 1.6
mM of Zn2+.
Conclusion: In this study, we reported a proteomic expression map of T. vaginalis grown in presence or absence
of Zn2+ and how the cytotoxicity and proteolytic activity of CP65 were affected by the presence of this cation in T.
vaginalis.
*Contact
Phone: (55) 5488-6661 ext.5353
e-mail: [email protected]
112
Science and Innovation Week 2009 • Abstracts Book
Healthy City
Obesity, diabetes and cardiovascular diseases
Genetic characterization of the polymorphisms T759T and E23K
of the genes SUR-1 and KIR6.2 in Mexican patients with DM2
Aguilar Juárez M1, Domínguez López A1, 2, Albor Valdés Z1, Cisneros González N3, Gómez López M1,
Arellano Flores ML3, Martínez Godínez MA1, Salgado Rodríguez J 4, González Mundo I4, López Sánchez
DM4, Delgado Hernández MA4, Arnaud Viñas MR4, Ramírez Campuzano R4, Miliar García A1*
1Escuela Superior de Medicina, Instituto Politécnico Nacional, 2Departamento de Gastroenterología,
Instituto Nacional de Ciencias Medicas y Nutrición Salvador Zubirán, 3Instituto Mexicano del Seguro Social,
4Subdirección Médica, Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado, México, D.F.
Abstract
Objective: The purpose of this study was to analyze the clinical characteristics of patients with DM2 in the first level
of attention of some states of the Mexican republic including population of the Mexico City and to determine the
presence of the polymorphisms T759T, of the gene SUR1 and E23K of the gene Kir6.2 and to observe if they have
association with the diabetic phenotype.
Research design and methods: Subjects with Type II diabetes (n=504) and normoglycaemic subjects (n=100) were
randomly selected from Mexican Institute of Social Security (IMSS). DNA was extracted from peripheral blood. Two
previously described SNPs in the sulphonylurea receptor gene in exon 18 T759T (ACC ACT) and inwardly rectifying
potassium channel gene in exon 1 E23K (AAG → GAG) were examined by a real time PCR.
Results: Statistically significant differences were not observed between the presence of SUR-1 with the genotype
of type heterozygous and homozygous in consideration towards the normality to the levels of glucose (OR 0.9,
95%CI), HbA (OR 0.66, 95% IC), cholesterol (OR 0.55, 95% IC) and triglycerides (OR 0.58, 95% IC) with risk for
alteration of the IMC (OR 2.17, 95% IC). When we analyzed Kir6.2, a significant difference in relation to levels of
decompensation for the studied variables was observed.
Conclusions: The genotypes of SUR-1 present acceptable levels in comparison with Kir6.2 that presents risk for the
decompensation of the levels in the five variables studied. Considering the previous thing it is possible to thing that
when one presents a polymorphism in SUR-1 considered as protective factor, does not appear in Kir6.2.
*Contact
Phone: (55) 5729-6000 Ext. 62768
e-mail: [email protected]
113
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Obesity, diabetes and cardiovascular diseases
Development of nanostructured materials for encapsulation of pentoxifiline
as coadjuvant treatment of diabetic foot
Álvarez M1*, López T 1,2,3, Jardón G 1,2, Ramírez P1,2, Ramírez Y 1,2, Rosas P 1,2
1Laboratorio de Nanotecnología, Instituto Nacional de Neurología y Neurocirugía M.V.S, 2Departamento de
Atención a la Salud, Universidad Autónoma Metropolitana, México, D.F. 3Department of Chemical Engineering,
Tulane University, New Orleans, LA, USA.
Abstract
Diabetic Mellitus (DM) is a group of metabolic diseases characterized by high blood sugar levels, as a result of
defects in insulin secretion or action. Over time, diabetes can lead to blindness, kidney failure, and nerve damage.
These are the result of damage to small vessels, referred to as microvascular disease. Diabetes is also an important
factor in accelerating the hardening and narrowing of the arteries (atherosclerosis), leading to strokes, coronary
heart disease, and other large blood vessel diseases. This is referred to as macrovascular disease. Diabetes affects
approximately 17 million people (about 8% of the population) in the United States. In Mexico, there are ca. 6.5
millions of patients with DM and about 50% will develop some type of neuropathy. Since 2003, this disease is the
main cause of death in this country.
Diabetic neuropathies are a family of nerve disorders caused by diabetes. People with diabetes can, over time,
develop nerve damage throughout the body. Nerve problems can occur in every organ system, including the
digestive tract, heart, and sex organs. About 60 to 70 % of people with diabetes have some form of neuropathy.
Peripheral neuropathy, also called distal symmetric neuropathy or sensorimotor neuropathy, most commonly affects
the feet and legs. Nerve damage in the feet can result in a loss of foot sensation, increasing your risk of foot problems.
Injuries and sores on the feet may go unrecognized due to lack of sensation. The prognosis for diabetic neuropathy
depends largely on how well the underlying condition of diabetes is handled. Treating diabetes may halt progression
and improve symptoms of the neuropathy, but recovery is slow. The painful sensations of diabetic neuropathy may
become severe enough to cause depression in some patients.
Foot infections in patients with diabetes cause substantial morbidity and frequent visits to health care professionals
and may lead to amputation of a lower extremity. The goal of treating diabetic neuropathy is to prevent further tissue
damage and relieve discomfort. The first step is to bring blood sugar levels under control by diet and medication.
Another important part of treatment involves taking special care of the feet. Analgesics, low doses of antidepressants,
and some anticonvulsant medications may be prescribed for relief of pain, burning, or tingling.
As a public health problem, is important to take steps for prevention, and at the same time to search alternatives to
diminish symptoms and accelerate injuries healing.
In this sense, with the use of nanotechnology, we can design suitable nanomaterials that allow us a local delivery
of drugs. With these systems, a sustained and controlled release of drugs is possible, decreasing adverse effects
and cost related with the use of large amounts of medicaments. In the present project, we prepared nanoparticles of
biocompatible inorganic metal oxide (SiO2) loaded with pentoxifiline, to incorporate them in a carbon wood to be
used as coadyuvant in the treatment of diabetic foot. The carbon wood would be bifunctional: as an adsorbent and
as a drug delivery system.
With this development, we expect to impact directly in the quality of life of patient with this affection.
*Contact
Phone: (55) 5606-3822 ext. 5034
e-mail: [email protected]
114
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Obesity, diabetes and cardiovascular diseases
Family study of genes associated with risk to develop conducts
related with eating disorders
Camarena B1*, Caballero A1, González L1, Hernández S1, Fresán A1, Alcocer N2
1Instituto Nacional de Psiquiatría Ramón de la Fuente, 2 Instituto Nacional de Ciencias Médicas y Nutrición
Salvador Zubirán, México, D.F.
Abstract
Evidence from family and twin studies suggests a genetic contribution in the etiology of eating disorders (ED). ED
is classified in Bulimia Nervosa, Anorexia Nervosa and Eating Disorders Not otherwise Specified (EDNOS). ED is a
major health problem in México City.
Clinical and pharmacological data suggest a dysfunction in serotonin system. In particular, the serotonin transporter
gene (SLC6A4) is a good candidate for genetic studies. A functional polymorphism (5-HTTLPR) located on the
promoter region has been associated with basal transcriptional activity levels. In addition, the 5-HT1Dβ is involved in
the regulation of 5-HT release from serotoninergic neuron terminals in the brain. Therefore, abnormal function of this
auto-receptor could be involved in the serotonin dysfunction suggested in ED. Also, it have been reported several
clinical characteristics that could be involved with abnormal food intake. For example, it have been reported a high
frequency of impulsivity, personality traits and anxiety in patients with ED.
This study proposes the collection of samples to create a DNA bank to identify the genes associated with clinical
characteristics involved in eating disorders in Mexican population.
The main objective of this study is to analyze the genetic variants and clinical characteristics associated with clinical
characteristic involved in the etiology of eating disorders. We will study the transmission of genes from parents to
affected sib using a family-based association method.
The sample is currently been captured from Eating Disorders Clinic of Instituto Nacional de Psiquiatría Ramón de
la Fuente. Probands will be interviewed by trained psychiatrist specialized in Eating Disorders to confirm the Eating
Disorder diagnosis by SCID-I interview (Spanish translation) according to the research criteria in DSM-IV TR. Also,
the patients will be assessment for impulsivity, anxiety, aggressive conduct, personality traits, obsessive-compulsive
traits and depression.
Several studies have been identified genes associated to traits in other populations. Therefore, it is important to
described the clinical characteristics in Mexican populations and detect the genes involved in the development of
ED.
The study will help to detect genes of risk that in the future could prevent conducts that may lead to develop an ED
in Mexican population.
Initiatives focused on identification of gene variants would greatly facilitate post-genomic research on the links
between genes, brain, behaviour and treatment response in Mexican population.
*Contact
Phone: (55) 5655-2811 ext.123
e-mail: [email protected]
115
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Obesity, diabetes and cardiovascular diseases
Clinical and experimental analysis of the impact of pre- and postnatal
undernourishment on the cardiovascular function and their impact
on the development of obesity and arterial hypertension
Hong E1*, Huang F2, Echeverría O1, Alvarado CE1, Bolaños J3
1Centro de Investigación y de Estudios Avanzados- Instituto Politécnico Nacional, 2Hospital Infantil de México
Federico Gómez, México, D.F 3UMR 1280 Physiologie des adaptations Nutritionelles, Nantes, France.
Abstract
Malnutrition is still a big problem of public health in Mexico despite the numerous national programs implemented
to eradicate it. An additional growing problem is the increasing obesity not only in adults, but also in infants. It
has been reported that children born from mothers with undernourishment during pregnancy frequently develop
obesity and/or arterial hypertension, but there is no explanation for this phenomenon. One possibility is that the
undernourishment period of the pregnant women produces a change in the metabolic programmation in the
children. This seems to happen in both, animals and humans. However, the mechanism of this phenomenon remains
unknown, and we intent to investigate such mechanism in the present Project.
In the clinical work, it will be investigated if children with obesity become from undernourished mothers. If this is the
case, blood samples will be taken in order to measure several possible markers, as well as to carry out an epigenetic
study in leucocytes.
In the work with animals, two female rats will be placed in contact with a male rat and after diagnosing pregnancy;
the female rat will be restricted to 50 % of food and let the pregnancy until it ends with a litter. These just born rats
will be compared with normal litters. In order to accelerate the process, the two types of rats will be fed with a special
pellet containing a greater amount of fat and sugar. The spectancy is that animals from undernourished mothers
will became fatter and hypertensive in relation to control animals. We also expect that they will suffer of endothelial
dysfunction, as well as a lack of the modulator effect of periarterial fat tissue on the vasoconstriction elicited by
several agonists.
The principal aim of this Project is to find out what drug would be able to prevent or to modify the effects induced
by undernourishment of pregnant females. Detection of biomarkers on obese children from mothers suffering from
undernourishment could be very useful to establish early treatments to prevent obesity and/or hypertension.
*Contact
Phone: (55) 5483-2864
e-mail: [email protected]
116
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Obesity, diabetes and cardiovascular diseases
Metabolic syndrome identification and psychological impairment in a
student population of Mexico City Autonomous University, and their
possible effects on academic performance
Jiménez Flores JR1, Murguía Romero M2, Rodríguez Soriano NY1, Villalobos Molina R4*, Méndez Cruz AR1
1Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, 2Unidad de Biotecnología
y Prototipos, 4Unidad de Biomedicina, Facultad de Estudios Superiores Iztacala, UNAM. México, D.F.
Abstract
Metabolic Syndrome (MS) is a pathological condition that includes several diseases (like: hyperglycemia, dyslipidemia,
hypertension, hyperuricemia, abdominal obesity, insulin resistance), which are among the 3 main causes of death
in México, according to the Ministry of Health. Previous reports show a prevalence of 26.3 % of MS in the Mexican
population between 20-69 years of age using the NCEP ATPIII criteria, and of 13.6% according to WHO. In the
Faculty of Higher Studies Iztacala (FESI) from the National University of México, we conducted a study with 973
rookies (30% of the new class, just finished High School), and we have found 14% hyperglycemic, 12% with high
total cholesterol, 42.7% with low HDL, 9% with high LDL, 17% with high TG, and 10% with hyperuricemia1. In
a psychological test ran in parallel, moderate to severe anxiety was observed in 74%, depression in 71%, low self
concept in 77%, risky to dangerous life style in 97%, inability in social affairs in 11%, deficient study activities in
20% of our students (17-20 years-old)1.
Despite those numbers, there is a lack of clinical signs that permits early MS identification then, our goal is to search
for early biochemical, molecular and psychological parameters to identify MS in a young student population, and to
relate those conditions with academic performance.
A 1000 student population will be recruited from the new class of this and next scholar years from México City
Autonomous University, a signed consent will be obtained to participate in the study. Students will be subjected
to a) clinical history and anthropometric measurements, such as: weight, height, waist, hip, waist-hip index, BWI,
blood pressure; b) in a blood sample: glucose, creatinine, uric acid, total cholesterol, triglycerides, HDL, LDL,
glutamic-pyruvic, glutamic- oxalacetic transaminases, -glutamil transpeptidase, serum iron, and albumin),and an
urine general study; c) a psychological test to evaluate anxiety, depression, study activities, self concept, life styles,
and social performance; d) in those students with MS signs, inflammation molecules will be determined in serum:
CD40L, ICAM-1, VCAM-1, E-Selectin, P-Selectin, and C reactive protein; e) those students with metabolic and
psychological impairment will be sent to specialized health centers; f) a software will be created to analyze the
data.
*Contact
Phone: (55) 5623-1108
e-mail: [email protected]
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Obesity, diabetes and cardiovascular diseases
TNF
, adipokines and insulin resistance as an initial stage
of the metabolic syndrome (MS)
Moreno Méndez E1, Vélez del Valle C1, Kuri Harcuch W1*
1Departamento de Biología Molecular, Centro de Investigación y de Estudios Avanzados -Instituto Politécnico
Nacional, México, D.F.
Abstract
Obesity and its associated diseases, such as MS are a public health problem in most parts of the world. The MS is a
set of metabolic abnormalities including obesity, insulin resistance, hypertension and dyslipidemia. This syndrome
results in the onset of cardiovascular disease, type 2 diabetes and some cancers, whose origin is obesity exacerbated
by the growth of white adipose tissue. The adipose tissue is an organ with the capacity to synthesize and secrete a
variety of molecules and proteins with endocrine, paracrine and autocrine activities causing physiological changes
in the body resulting in the pathophysiology of MS and subsequent diseases. Obesity is associated with white
adipose tissue and its pro-inflammatory state. This inflammation is due to a chronic activation of the innate immune
system that leads to a phenomenon of insulin resistance and other metabolic disorders, accounting for MS. Among
the key inflammatory molecules produced by adipose tissue are: tumor necrosis factor alpha (TNF- ), the monocyte
chemotactic protein 1 (MCP-1) and interleukin-6 (IL-6). These inflammatory molecules, in addition to local effects
in adipose tissue, also exert their effects at a systemic level in tissues primarily related to metabolism, storage and
consumption of energy such as: cardiac and skeletal muscle, islets of Langerhans, liver and the adipocyte itself,
among others. There have been several studies regarding the effect of TNF- in the biology of adipocytes, but it has
not been clearly established that this cytokine is produced by fat cells. In our laboratory, using a system of adipose
differentiation which allows the study of adipocytes from induction of differentiation to the mature phenotype, we
explored the production of TNF- . So far our results show that the TNF- messenger RNA is present in cells from
preadipocytes to mature adipocyte.
*Contact
Phone: (55) 5747-3353
e-mail: [email protected] , [email protected]
119
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Obesity, diabetes and cardiovascular diseases
Epigenetics of cognition and neurophysiology in a twin sample from Mexico
City: potential endophenotypes for obesity and addiction
Nicolini H1*, Genis A1, Camarena B 1,2, Martínez A1, Meyenberg N 3, Ochoa M4, Santana D3, Lanzagorta N3
1Posgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de México, 2División de Investigaciones
Clínicas, Instituto Nacional de Psiquiatría, 3Grupo Médico Carracci, 4Jefatura de Enseñanza y Servicio de
Psiquiatría, Hospital 20 de Noviembre, Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado,
México, D.F.
Abstract
Introduction: Preliminary data will be presented from an ongoing study supported by the ICyTDF in the epigenetics
of cognition and neurophysiology in a monozygotic twin’s sample. Obesity and addictions are two major health
problems in Mexico City. In both behaviors there is extensive data published supporting their genetic basis. In addition
it seems that there are common genetic and phenomenological links between both conditions. These findings lead us
to the emerging concept of endophenotype which is a measurable component unseen by the unaided eye along the
pathway of disease and genotype (Gottesman et al., 2003). Establishing a particular measure as an endophenotype
requires the trait to be heritable, sensitive to affection status, and genetically correlated or associated with the illness.
The establishment of a particular trait as an endophenotype is most efficient with a twin’s sample, since we have two
genetically identical individuals which will differ depending upon their exposure to different environments. These
twin monozygotic siblings provide a single sample where all of the criteria for effective endophenotypes can be
assessed simultaneously (e.g. heritability, sensitivity to the illness, genetic correlation). Differences at the molecular
level on gene expression (purposely caused by environment) can be assessed as well by methylation.
Methods: This sample is currently been recruited from the registry of the Mexican National Association of Multiple
Births from Mexico City and other clinical sites. This is the first twin study been conducted in Mexico. We are assessing
all subjects with a full neurocognitive battery, evoked related potentials (P50, Mismatch Negativity, and P300), as
well as medical variables, BMI, glucose levels, cholesterol, lipids, test for drug abuse, expired CO, psychiatric and
psychometric clinical structured interviews. In addition, we are obtaining DNA samples and processing them by
bisulfite method to assess methylation. We will be evaluating the epigenetics of several candidate genes involved in
the vulnerability to obesity and addiction, two major health problems in Mexico City.
Preliminary results: We are presenting some examples of those measures that we have been obtained so far at all
phenotypic levels (cognitive, neurophysiologic, medical, etc.). These results will lead into future directions on the
effect of environment on candidate gene expression (epigenetics). Also, these findings may create the possibility to
detect and possibly to prevent those environmental risks that may lead to addiction and/or obesity in Mexico City.
Funding: Instituto de Ciencia y Tecnología del D.F. (ICyTDF)
*Contact
Phone: (55) 5850-1901 ext. 15305
e-mail: [email protected]
120
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Obesity, diabetes and cardiovascular diseases
Oxidative stress and insulin resistance in a model of metabolic
syndrome: effect of glycine
Ruiz A1, Chávez M1, Baños G1, El-Hafidi M1*
1Departamento de Biomedicina Cardiovascular, Instituto Nacional de Cardiología Ignacio Chávez, México, D.F.
Abstract
The metabolic syndrome (MS) is a cluster of several heterogeneous risk factors such as diabetes mellitus type II and
diverse cardiovascular diseases. The clinical manifestations of MS include insulin resistance, hypertriglyceridemia,
abdominal obesity and hypertension. In Mexico City and in the world, the exponential increase in the prevalence of
this pathology in recent years could be due to the changes in lifestyle that influence genes which regulate metabolic
mechanisms. In order to understand the multiple interactions between genetic, nutritional and environmental factors,
different strategies should be designed to prevent MS, such as healthy diets and exercise. Many patients for social
reasons and/or current lifestyle are not able to change their dietary regime nor the way they take exercise. Therefore
it is necessary to develop pharmacological or nutritional supplementary treatments, that do not have secondary
effects, thereby improving the quality of life of MS patients.
A rat model of MS has been developed in our laboratory, which represents the pathology resulting from a faulty life
style, with inadequate diets due to the excessive intake of carbohydrates.
Glycine, a non-essential amino acid, is required by a number of metabolic pathways, among them the synthesis of
structural proteins such as collagen and elastin. Glycine, when fed to MS rats, causes significant alterations in body
composition and metabolism. We reported that dietary glycine caused a significant decrease of intra-abdominal fat,
arterial blood pressure and resistance to insulin, in our rat model of metabolic syndrome. These results suggest that
glycine treatment has potential as an anti-obesity agent.
Therefore this project aims to elucidate the mechanism by which glycine affects insulin resistance through the
different signalling pathways induced by insulin. These pathways imply interactions among different proteins and
the translocation of the glucose transporter4 (Glut4). Glycine can increase the sensibility to insulin by several
mechanisms, including its interaction with its receptor, which activates the chloride channel and regulates intracellular
Ca2+. Glycine also participates in the biosynthesis of the reduced glutathione (GSH) catalyzed by two enzymes,
glutamylcysteine synthetase and GSH synthetase. The GSH participates in antioxidant defense as a substrate of
glutathione peroxidase. A diminished level of circulating glycine induces a subsequent deficiency of GSH and
an increase in the oxidative stress which is involved in insulin resistance. Preliminary results indicate that glycine
increases the sensitivity to insulin, by increasing the phosphorylation of its receptor IRbeta and of IRS-1 at tyrosine
residue, in liver of MS animals.
*Contact
Phone: (55) 5573-2911 ext.1237
e-mail: [email protected]
121
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Obesity, diabetes and cardiovascular diseases
Chronomic aspects of pre-metabolic syndrome (CPMS)
on Mexico City population
Sánchez de la Peña S1*, Halberg F2,Mendoza Lujambio I1,3, López Fiesco A4, Cornélissen G2,Rito Levi J1
1Centro de Investigación Cronómica, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional,
2Halberg Chronobiology Center, University of Minnesota, Mpls. MN, EUA, 3Genética Molecular, Escuela
Nacional de Medicina y Homeopatía-Instituto Politécnico Nacional.
4Clinica Revolución del Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado, México, D.F.
Abstract
Background: In the development of Metabolic Syndrome (MS) cardio and metabolic risk factors have been associated.
These entities had shown elevated prevalence and their progression of cardiovascular and renal diseases. Cardiometabolic syndrome (CMS) and SM shared a complex conjunction of diabetes, hypertension and obesity. The
medical care costs, that both syndromes induced at EUA had reached about 80 billion dollars. The clinical, social
and economical outcomes are very similar as in Latin-American countries. Mexican government health agencies
are actively developing preventive programs, related to prevent MS. However, mainly based upon linear medical
approach, as it has been developed all around the world, without consideration of daily and weekly variability of
vital signs, as blood pressure (BP). A considerable accumulated vast evidence on the human BP temporal structure
or chronome might modify some classical aspects of hypertension and MS. By studying the BP chronome on
normotensive people, it was identified a circadian vascular variability named “CHAT” (Circadian HyperAmplitude
Tension), and characterized also as hyperglycemic persons. Both chronomic and metabolic signs constitute a new
Chronomic Pre-Metabolic Syndrome (CPMS). This syndrome has been recently confirmed on black community.
Principal aim: to develop a Chronomic (interaction of endogenous and external biorhythmicity) study related to the
presence of CPMS and other Variable Vascular Disorders (VVDs) on different Mexican populations starting on Mexico
City. All these detected by automatic BP monitoring on 7-days/24 h and capillary blood glucose. These VVDs are:
a) MESOR-hypertension, b) BP overswinging or CHAT; c) excessive pulse pressure; d) odd timing of BP variability
named circadian rhythmic Ecphasia of BP and e) MESOR hypothension. Specific aims: to explore the presence of
three clock genes (hPer1, hCry1 y hBmal1) on subjects diagnosed with CPMS, Hypertension and VVDs.
Subjects and Methods: Following the criteria of good clinical practice and bioethical approaches, residents of
Mexico City as well as, other country cities of both genders will invited to participate in this study, by monitoring
their BP for span of 7 or more days and complemented with tolerance and fasting glucose tests. Detection of VVDs
and CPMS will be determined at CRC a the IPN and at the Halberg Chronobiology Center, University of Minnesota,
Mpls EUA.
Preliminar results: Rhythmometric statistical evaluation confirmed the presence of CPMS on residents of some cities
around Mexico City. Large double amplitude systolic BP has been identified on Mexico city residents. More detailed
information will be presented at the poster session.
Grant support by ICyTDF: PICDS08-82.
*Contact
e-mail: [email protected]
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Obesity, diabetes and cardiovascular diseases
Electrophysiological and neurochemical correlates of satiety states
in obese and lean subjects
Téllez L1, Lodoño M1, Obed I1, Bermudez RF2 , Ranier G1*
1Departamento de Farmacología, Centro de Investigación y de Estudios Avanzados-Instituto Politécnico Nacional,
2 Instituto de Fisiología Celular, Universidad Nacional Autónoma de México.
Abstract
The study of how the brain induces satiety is relevant for Mexico, considering that obesity is one of the most
growing health issues in the world. In 2006 a health survey revealed that 70% of adult mexican population has
overweight or obesity. Obesity potentially reduces life expectancy and it is a risk factor for cardiovascular disease,
certain types of cancers and development of diabetes type II. This research project has two main goals; first we
want to understand how the brain encodes food induced-satiety, and second how the intestinal peptide CCK-8 and
hormone leptin transfer its satiety signal to the brain in lean subjects, and then characterize how this satiety signals is
impaired during obesity. To achieve these goals, we are combining two state of the art technologies: microdialysiselectrophoresis capillarity and multi-electrode recordings. The first technology allows us to simultaneously measure,
minute by minute, the extracellular release of neurotransmitters such as dopamine, GABA and Glutamate. The
second one, by implanting micro-arrays of 32 electrodes (each electrode thinner than a human hair), we can record
the electrical activity of neuronal ensembles in the Nucleus Accumbens, while hungry rats feed themselves until
sated. The nucleus accumbens is a region of the brain that responds to rewards and nucleus accumbens neurons
release dopamine during sucrose intake until satiety. It has been shown that obese subjects showed a decrease on
dopaminergic transmission throughout D2 receptors, in the nucleus accumbens. Therefore, the nucleus accumbens
is not only important for food intake regulation, but also its malfunctioning seems to contribute in the development
of obesity. Our research will uncover for the first time, the electrical and neurochemical participation of the nucleus
accumbens in encoding satiety states as well as its potential role in the development of obesity.
*Contact
Phone: (55) 5747-3800 ext.5426
e-mail: [email protected]
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Obesity, diabetes and cardiovascular diseases
Insulin and nerve growth factor in obesity and metabolic syndrome
Velasco Torres M1, Larqué Velásquez CA1, Albarado Ibáñez A1, Martínez AL1, Sánchez SC1, Hiriart M1*
1Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, México, D.F.
Abstract
Metabolic syndrome (MS) is a multifactor complex of signs that increases the probability to develop several types of
health problems, cardiovascular processes, diabetes and certain types of cancer among them. Some of the principal
problems in MS are obesity, hypertension, hyperinsulinemia and high levels of serum triglycerides. Causes and
physiopathology of MS are not well understood. Clearly it has two types of components, genetic and environmental,
the later related mainly to diet and obesity.
We are interested in analyzing the specific effect of a high-sucrose diet on pancreatic islet morphology and
physiological parameters in an adult rat model.
Insulin produced and secreted in pancreatic beta cells is crucial for nutrient homeostasis in mammals. Beta cells are
derived from gut precursor stem cells, however it is not entirely clear how are islets formed during development and
how are beta cells renewed when pancreas is full developed and if new islets are formed in adult rats. Mechanisms
that participate in acquisition and conservation of beta phenotype are not clear yet. They may depend on growth
factors. Pancreatic beta cells produce and secrete nerve growth factor (NGF) and have high affinity receptors for
NGF in their membrane. We have previously observed that NGF increase glucose induced insulin secretion, partially
acting on sodium and calcium channels and membrane excitability in beta cells from newborn rats. We have also
observed that nearly birth time NGF may contribute to sympathetic innervation of islets.
Diets with high carbohydrate content during lactation can cause sustained hyperinsulinemia and obesity in adult
rats. Also alterations in islet size, number and composition have been observed in response to nutritional changes.
These changes may derive in a higher risk for type 2 diabetes development.
We developed a model of metabolic syndrome in adult male Wistar rats (MSR), by feeding them during 8 to 24 weeks
with a 20 % sucrose solution in drinking-water (w/w; MSR group, n = 23) vs. the control group, which received plain
water (n = 24). We are analyzing, body weight (BW), adipose tissue weight, plasmatic glucose, and arterial pressure
were measured; plasmatic insulin and cytokines produced by adipose tissue. As well as morphological changes of
pancreatic beta cells and adipose tissue.
After two months of treatment BW was 20% higher in MSR than in the control group; this was mainly due to the
increase in abdominal fat. At the end of six months of treatment, MSR preserved BW difference, by a 2.2-fold
increase in abdominal adiposity. At both stages, arterial pressure was 10% higher than in the controls and showed
hyperinsulinemia.
Preliminary results show that at 6 months of treatment, plasmatic insulin decreased by 31% compared to controls;
while glucose concentration tended to increase in MSR, without statistical significance. At 6 months, pancreatic
insulin area and immunolabeling were nearly 2.5-fold higher in MSR than in controls, without changes in glucagon
area or intensity. Our results suggest that in a long term exposure, high sucrose intake increases beta cell number;
however insulin-secretion in the new cells is impaired. These alterations may lead later to beta cell exhaustion
and type 2 diabetes mellitus. We are also interested in studying the reversibility of these changes and the female
metabolic behavior.
*Contact
Phone: (55) 5622-5665
e-mail: [email protected] , [email protected]
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Obesity, diabetes and cardiovascular diseases
Mutations in the GCK and GLUT-4 genes and leptin and adiponectin levels
in overweight, obese and diabetic-obese Mexican patients
Vergara Calderón N1, Flores Velázquez R3, Rangel Rivera M3, Audelo Chícharo G2, Lara Padilla E1,
Mendoza Alcántar L1*
1Sección de Estudios de Posgrado e Investigación, Escuela Superior de Medicina del Instituto Politécnico
Nacional, 2 Estudiante de posgrado del Instituto Politécnico Nacional,
3Estudiante PIFI del Instituto Politécnico Nacional, México, D.F.
Abstract
Objective: To analyze mutations on GCK and GLUT-4 genes and serum levels of leptin and adiponectin in overweight,
obese and diabetic-obese patients.
Introduction: Obesity is a “pandemic illness” in the world affecting 300 millions of individuals. In México
approximately 70 % adults have overweight u obesity and is placed in 2nd position after USA, due this is considered
a public health problem. Different reports have demonstrated association of mutations on genes related with obesity
and type 2 diabetes mellitus (T2DM) diseases. We began this study to identify mutations on GCK and GLUT-4 (type
2 diabetes-related genes) and leptin and adiponectin serum levels (obesity-related genes) in Mexican patients.
Material and methods: Obese, diabetic-obese and overweight patients (total 200) were included in the study. Age
was between 18-77 years old and anthropometric measures to calculate body mass index (>25 kg/m2) were made.
Blood samples were obtained to separate serum and white cells. Serum was used for determine biochemical profile
including; blood glucose levels, HDL-c and triglycerides. Leptin and adiponectin determinations were made using
commercials kits by ELISA. Leukocytes were processed for DNA isolation and amplification by PCR of the GCK and
GLUT-4 genes for sequencing.
Results: The 72 % were obese and diabetic-obese patients and 28 % overweight individuals. High serum levels of
leptin were associated only with obese (p≤ 0.001), but not with diabetic-obese subjects. Interestingly, overweight
patients showed also elevated concentrations of leptin. On the other hand, adiponectin was detected in low levels
in all patients. Genomic DNA was isolated from the 200 samples and used to amplify by PCR the DNA fragment of
GCK and GLUT-4 genes which are under sequencing.
Discussion: Changes in leptin and adiponectin serum concentrations have been reported by different studies in USA,
European and South of America obese population. Frequently, these modifications are due mutations located in
the regulatory or encoding region of these genes. We found similar results in the seric leptin and adiponectin levels
in obese Mexican patients. Interestingly, high leptin concentration in overweight individuals, indicate high risk to
develop obesity and its complications. We are looking for association between mutations on GCK and GLUT-4 genes
and serum leptin and adiponectin levels in our population.
This research has been funded by Instituto de Ciencia y Tecnología del DF and by Instituto Politécnico Nacional,
México.
*Contact
Phone: (55) 5729-6000 ext. 62793, 62746, 62743, 62806.
e-mail: [email protected]
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Mental health and addictive behaviors
Genetic expression levels of 5-HTT and IFN-y in major depression
patients along 52 week of follow-up with SSRI
Becerril LE1, 3, Hernández ME1, Castañon M3, Martínez M3, Mendieta D2, Alvarez L1, Pavón L1*
1Departamento de Psicoinmunología, 2Servicios Clínicos del Instituto Nacional de Psiquiatría, 3Posgrado en
Ciencias Genómicas, Universidad Autónoma de la Ciudad de México, México, D.F.
Abstract
Introduction: In Major Depressive Disorder (MDD) there is overactivity in the hypothalamus-pituitary-adrenals (HPA)
axis. This fact is associated to altered levels in soluble mediators from nervous system, such as serotonin (5-HT);
from the endocrine system like the cortisol, and interferon-gamma (IFN-) from the immunologic system. These
variations affect peripheral blood mononuclear cells (PBMC) like the lymphocytes since they consecutively express
receptors for glucocorticoids, cytokines and serotonin (5-HT1A and 5-HT2 and 5-HTT). Additionally, it has been
shown that peripherical variations in the levels of cytokines in MDD during treatment with selective serotonin
recapture inhibitors (SSRI) are a consequence of genetic modulation. These facts suggest that genetic variations in
lymphocytes can be used as markers of treatment efficacy in MDD. However little is known regarding 5-HTT and
IFN- genic expression during chronic treatment with SSRI in MDD.
The purpose of this study was to determine by PCR-real time (RT) the expression levels of 5-HTT and IFN- from
PBMC in MDD patients along 52 weeks (W) of follow-up with SSRI. Twenty healthy volunteers (HV) and thirty MDD
patients who were treated with SSRI participated. 5-HTT and IFN- by PCR-RT genic expression determinations were
performed at week (W) 0, 5, 20, 36 and 52. Our results prove that, when compared to HV, there is a decrease in the
expression of 5-HTT in MDD patients before the treatment. The acute administration of SSRI during the first 20 weeks
increases the genic expression of 5-HTT. However expression levels of 5-HTT similar to W0 were detected from W36
and until the end of the study. In regard to the expression of INF-, an increase in MDD patients was found in W0
when compared to HV. However, there was a decrease in W20 and from W36 to the end of the follow-up the IFN-
expression was similar to that detected in W0.
Discussion and conclusion: In relation to knowledge about the genetic regulation that is presented in MDD patients
during SSRI treatment, currently is limited. Our results suggest a partial restoration of 5-HTT genic expression and
IFN- with SSRI treatment during the first 20 weeks, even though the mechanisms related to immune modulator
effects of SSRI are not yet established. We believe that the SSRI lead to a partial restoration of 5-HTT expression,
which favors the reduction of free 5-HT. At the same time this fact induces the decrease in the expression of INF-.
Further studies are needed to explore the clinical use of these findings and their likely therapeutic application in
MDD.
Financing: This study was supported by the Instituto de Ciencia y Tecnología del Distrito Federal (ICyTDF-2007) and
INPRF-NC092318.1
*Contact
Phone: (55) 4160-5082
e-mail: [email protected]
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Mental health and addictive behaviors
Individuals with prodromal symptoms for schizophrenia showed abnormal
brain activation to anticipation of unpleasant stimuli: an FMRI study
De la Fuente Sandoval C1*, Favila R 2, Graff Guerrero M 3, León Ortiz P4, Fresan A 5, Stephano S 1, Menon M 6,
Graff Guerrero A6
1 Laboratorio de Psiquiatría Experimental, Instituto Nacional de Neurología y Neurocirugía, México, D.F,
2 MR Advanced Applications, GE Healthcare. 3 University of Essex, Colchester, UK, 4 Departamento de
Neuropsiquiatría, Instituto Nacional de Neurología y Neurocirugía, 5 División de Investigación Clínica, Instituto
Nacional de Psiquiatría. México, D.F, 6 Department of Psychiatry, University of Toronto & Centre for Addiction
and Mental. Toronto, Canada.
Abstract
Background: Schizophrenia is a mental disease with a general population prevalence of 1%, and affects 50 million
people around the world. The World Health Organization has classified Schizophrenia as a public health problem;
it represents the eighth cause of disability in the world. In Mexico, it is estimated that 1 million persons are affected
by this disease, and that 20% of the patients with schizophrenia are severely incapacitated.
Traditionally, this disease is characterized by two groups of symptoms: the positives and negatives, from which the
positives or psychotic are fundamental for its diagnosis. These symptoms are characterized by a lost of contact with
reality, incoherent o illogical ideas (delusions), hallucinations (perception experiences without a sensorial stimuli)
or behavior disturbances (such as bizarre conduct or aggression). Psychotic symptoms can be seen in most of the
patients as the disease begins and their manifestation and severity tends to be episodic along with time.
It is thought that the formation of abnormal conditioned associations might be related to the formation of symptoms.
Prior work has also demonstrated that associative learning during aversive conditioning tasks is abnormal in
schizophrenia. However, this abnormality has not been described in individuals with an enhanced risk to develop
the disease (with prodromal schizophrenia symptoms). We hypothesized that individuals with prodromal symptoms)
would exhibit abnormal brain functioning during the anticipation of unpleasant stimuli.
Methods: Fifteen participants that met Structured Interview for Prodromal Syndromes criteria (12 accounted for
Genetic Risk) and 15 healthy controls paired by age, sex and handedness were included. High field functional
Magnetic Resonance Image - fMRI (3 Tesla) were used to measure the blood oxygen level dependent (BOLD) signal
while they carried out an aversive Pavlovian conditioning task with 33% reinforcement, using left index finger
electrical stimulation as aversive stimulus and red and yellow circles as visual cues.
Results: Random effects analyses (second level) were used to identify brain regions that were more active in response
to the conditioned stimuli relative to the neutral stimuli in the two groups. The group comparison revealed that the
prodromal participants showed greater activity than control group in the limbic and paralimbic regions-including the
medial temporal cortex, thalamus and insula during anticipation the aversive stimulus.
Conclusions: Our results suggest that prodromal subjects show an atypical and heightened neural activity in limbic
regions in the anticipation of aversive stimuli.
The activated regions correspond to brain processing of the aversive stimulus itself and not to anticipation. This
may contribute to the aberrant assignment of salience to neutral stimuli which may in turn, predispose them to
psychosis.
*Contact
Phone: (55) 5606-3822 ext.1034
e-mail: [email protected]
130
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Mental health and addictive behaviors
Expression of proteins for gene therapy by using recombinant
adeno associated virus
Juárez Galicia D1, Vázquez Carcaño LM1, Uribe M1, Arias CF2, Palomares LA2, Torres Vega MA1*
1Departamento de Gastroenterología, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán,
México, D.F. 2Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos.
Abstract
Gene therapy is a strategy that allows us to introduce genes into the cells of an organism to express one or several
proteins, with the goal of correcting a deficiency or a disease. We would like to establish this strategy in the INCMNSZ,
in order to offer new alternatives to the patients who arrive at this hospital. Recombinant adeno associated virus
(rAAV) is currently used in more than 40 gene therapy clinical protocols. The rAAV virus is safe, it does not cause
any pathology in humans and it triggers a very low immune response. The rAAV expresses high levels of therapeutic
proteins during long time (one year or more), and it can be produced with a very high title. At the beginning, we want
to express therapeutic proteins in several tissues. For this purpose, we will use different AAV serotypes, which exhibit
a specific tropism for each tissue. Now, we are subcloning the EGFP cDNA in the viral vector. Later, we will clone the
cDNAs of therapeutic proteins in this vector. The viral particles with the EGFP or therapeutic cDNAs inserted in their
genome will be produced in cells in culture. Purified rAAV virions will be injected intraperitoneally or specifically in
one organ or tissue. Furthermore, we will check the expression and/or the activity of the EGFP reporter or therapeutic
proteins in the whole organism or tissue samples.
This project is supported by the Science and Technology Institute from Mexico City (ICyTDF).
*Contact
Phone: (55) 5673-3902
e-mail: [email protected]
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Mental health and addictive behaviors
Role of the endogenous opioid system in alcohol dependence
Méndez M1*, Hernández Fonseca K1, Martinell P1, García F1, Pérez Luna JM1, Barbosa Luna I1, Herrera S,
Munguía A1, Morán J2
1 Departamento de Neuroquímica, Instituto Nacional de Psiquiatría Ramón de la Fuente, 2 Departamento de
Neurociencias, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México.
Abstract
Alcoholism is one of the main health problems over the world. Alcohol abuse and dependence seriously affect
the function of several organs in the body, including the brain. Important alterations in brain function have been
described after high alcohol consumption, which could be attributed to neuronal damage and cell death.
Alcohol (ethanol) exerts its actions through neural reinforcement mechanisms, which involve activation of the
dopaminergic mesolimbic system. Besides dopamine, other neurotransmitters and neuromodulators may be involved,
including opioid peptides. Neurochemical and pharmacological evidence suggest that the reinforcing effects of
alcohol are mediated, at least in part, by the ethanol-induced activation of the endogenous opioid system. Several
studies support this view. Thus, opioidergic systems are important neural targets of ethanol’s actions, although their
role in alcohol dependence mechanisms remains to be established.
Behavioural sensitization to drugs of abuse has been recently established as an animal model of drug dependence.
Behavioural sensitization is induced by the repeated administration of psychoactive substances, and is characterized
by a progressive increase in the locomotor response, as well as the persistent hypersensitivity to the stimulant effects
of drugs. Neuroadaptive mechanisms may be progressively established during this process, which could lead to drug
dependence. Behavioural sensitization to several psychoactive substances has been reported, including cocaine,
amphetamine, heroin and alcohol.
Alcohol behavioual sensitization has been difficult to investigate, due to the biphasic effects of the drug. Low doses
of ethanol induce psychomotor activation and euphoria, whereas high doses decrease motor activity and produce
sedation. Ethanol doses that induce locomotor activation in rodents have been difficult to assess. In addition, the
stimulant effects of ethanol are influenced by several factors, including the route and duration of drug administration,
light-dark cycle, rodent strain and sensitivity to stressful stimuli (isolation, novelty, etc). Thus, the influence of these
factors must be considered when studying ethanol effects.
The dopaminergic mesocorticolimbic and nigrostriatal pathways play a key role in behavioural sensitization to
alcohol and other drugs of abuse. Other neurotransmitters and neuromodulators, such as glutamate and opioid
peptides could play a key role in alcohol behavioural sensitization. Thus, the aim of this research project is to
investigate the role of opioid peptides (enkephalins and ß-endorphin) in the mechanisms that take place during
alcohol dependence, using behavioural sensitization in Wistar rats as experimental model. We will correlate alcohol
motor and anxiolytic effects with the ethanol-induced changes in opioidergic transmission in brain areas of the
mesocorticolimbic and nigrostriatal pathways in sensitized rats. In addition, we will determine if the repeated
administration of ethanol induces neuronal death in these brain areas, since the prolonged exposure to ethanol is
known to produce significant neurotoxic effects, both in humans and animals.
*Contact
Phone: (55) 5655-2811 ext. 212
e-mail: [email protected]
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Multicentre study of the polymorphisms in the codificant regions
of the genes associated to the drug resistance in pediatric patients
with focal epilepsy
Orozco Suárez S1*, Feria Romero I1, Grijalva Otero I1, García Ramírez R2, Rayo Mares D2, Fraire Martínez MI3,
Pérez Ramírez JD 4, Ramírez Reyes G4, Cabrera R5, Sosa Maldonado J6, Ruiz Chávez J6
1Unidad de Investigación Médica en Enfermedades Neurológicas, Hospital de Especialidades, 2Departamento de
Neurología, 3Neurofisiología, 4Neurocirugía, 5Salud Mental, Hospital de Pediatría, Centro Médico Nacional Siglo
XXI, 6Neurología Pediátrica, Hospital de Especialidades, CMN La Raza, IMSS, México, D.F.
Abstract
Introduction: Epilepsy is a neurological disorder characterized by recurrent seizures that affects approximately
1.8–2% of the Mexican population. Despite considerable advances in the pharmacological treatment of epilepsy,
approximately one-third of epileptic patients are pharmacoresistant. At molecular level the factors that are associated
to refractory are: Genome variability (for example, gene polymorphisms leading to alterations in drug metabolism,
drug targets or drug transporters). Such variability might explain why two patients with the same disease differ in their
initial response to drug treatment. Disease-related mechanisms. These might include the aetiology of the disease,
disease progression despite treatment, structural brain alterations and/or network changes, alterations in drug target(s)
or alterations in drug uptake into the brain. Drug-related mechanisms, such as loss of therapeutic efficacy; induction
of drug-metabolizing enzymes or drug transporters; and ineffective mechanisms of drug action. In Mexico it is not
known the association between the polymorphisms of the candidate genes (molecular target, metabolize enzymes and
multidrug transporters proteins), with the therapeutic range of AEDs in the pharmacoresistant patients. In the present
work we study the association between the plasmatic levels of new AEDs and polymorphisms of genes SCN1A,
SCNÁ, ABCB1, CYP2C9, CYP2C19 and CYPÁ4 in pediatric patients with focal epilepsy and pharmacoresistant
Objective: Identify the polymorphisms of genes associated to the drug resistance in pediatric patients with complex
partial epilepsy resistant to pharmacological treatment.
Methods: For this study we included patients with resistant and responsive epilepsy to the pharmacological treatment,
with a demonstrable and classified epileptic focus, and healthy volunteers of the different departments of pediatric
neurology of the participants hospitals. The identification of resistant patients will become by the quantification the
therapeutic maxim range of DAEs. The allelic frequencies and the genotypes associated with the resistance to the
drug will be obtained by the specific amplification of exons 1, 12, 21 and 26 of gene ABCB1, exons, 7,9,10,16,29,33
y 34 of gen CANAIH, exons 5,10,15,16 y 26 of gen SCN1A, SCN3A, exons 3, 7 y 8 of gen CYP2C9, exons 1, 2, 3
y 7 of gen CYP2C19 and exons 6 and 12 of gen CYP3A4. The genomic DNA is obtained of the peripheral blood
of the patients and volunteers. The amplified fragments will be quantified; sequence and they will be analyzed
statistically to determine the association parameters. We obtain results by amplified and sequence exons 5, 10, 15,
16, 23 and 26 of gene SCN1A, the sodium channel, that results of the sequences and the genetic segregation have
been analyzed in 5 families. The methods were standardized of quantified; Carbamazepine, levetiracetam, and
oxcarbazepine in serum by hi-resolution chromatography (HPLC).
*Contact
Phone: (55) 5578-0240
e-mail: [email protected]
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Structural studies of sol-gel silica-dopamine biomaterials for used
as drug delivery reservoirs in the central nervous system
Ortiz Islas E1*, Lopez T1,2,5, Esquivel D3, Quintana P4, Oskam G4, Ascencio J6
1 Laboratorio de Nanotecnología, Instituto Nacional de Neurología y Neurocirugía,
2 Departamento de Atención a la Salud, Universidad Autónoma Metropolitana-Xochimilco, 3 Centro de
Investigaciones en Química Inorgánica, Universidad de Guanajuato, 4 Departamento de Física Aplicada,
Centro de Investigación y de Estudios Avanzados (Unidad Mérida) del Instituto Politécnico Nacional. México,
5Department of chemical and Biomolecular Engieneering, Tulane University. New Orleans, USA. 6 Instituto de
Ciencias Físicas, Universidad Nacional Autónoma de México. Cuernavaca, Morelos. México.
Abstract
Sol-gel silica-dopamine biomaterials were prepared in order to obtain a stabilized dopamine into functionalized
delivery reservoir. Dopamine is a neurotransmitter generated in dopaminergic neurons in the substantia nigra pars
compacta region of the brain. Massive loss of dopamine in the nuclei of caudate and putamen leads to Parkinson
disease. The common treatment for this disease is the oral administration with L-DOPA, however, improve
parkinsonian symptoms but do not repair the dopaminergic pathway or prevent its degeneration. Prolonged use of
L-DOPA develops tolerance and severe side effects. The nanomedicine is a good alternative to resolve this problem.
We propose a new strategy, the use of an implantable nanodevice to liberate dopamine during one year in the
caudate nucleus. Thus, is possible to maintained the dopamine pure and avoid the oxidation to any quinone, as
well as decrease the dosage and to make the drug available at the target site. To know the diffusion into the brain we
simulate a viscose system (agar with cefalorraquideum liquid) to study the trajectory of the chromophore-dopamine
“in vitro”. For this purpose, dopamine was added into the silica network during the TEOS hydrolysis step using
several functional groups. The SiO2-Dopamine reservoirs obtained were characterized by N2 adsorption, TEM
microscopy, FTIR spectroscopy and the “in situ” release profiles were make. In the other hand, a theoretical study
was development with the purpose to determine the possible modifications in dopamine electronic structure as a
result of the presence of H atoms with SiO2 units. The textural analyses showed a maximum value of surface area
(620 m2/g) for silica with 19.2 % wt of dopamine in the mesoporous nanostructured solids. The stability of dopamine
into the network silica was probe by FTIR, Raman and MNR spectroscopies. The bands localized at 2500, 2600 and
2700 cm-1 are attributed to N-H, C-H and C-O-H vibrations of dopamine, indicative that the structure of dopamine
remains intact. The “in vitro” Dopamine delivery profiles indicate two delivery steps. A fast-sustained dopamine
delivery was observed until 24 hours, after this time the delivery is constant. The theoretical studies indicate that
dopamine is attracted by the silica in coplanar form. Nevertheless, the calculus indicated that dopamine is linked to
silica through silanol groups and amine groups of the dopamine. This interaction stabilizes dopamine into of silica
network.
*Contact
Phone: (55) 5606-3822 ext.5034
e-mail: [email protected] , [email protected]
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Psychological, Genetic and Neurocognitive Correlates
Of Violence in Humans
Ostrosky SF1*, Bobes MA2,Trejo MD3, Romero C 1,Borja K1,,Díaz K1, Salgado JC 1, Medina BV4
1Laboratorio de Psicofisiología y Neuropsicología, Facultad de Psicología, Universidad Nacional Autónoma de
México, 2Centro de Neurociencias de Cuba, La Habana, Cuba. 3Hospital General de México, 4Universidad
Autónoma Metropolitana-Iztapalapa, México, D.F.
Abstract
Violent behaviors are alarmingly common in our society and regarded as a public health problem. It ranges from
domestic violence to murder and crime in the streets. Research aiming at understanding aggression and violence
as well as its causes has increased in an effort to combat this issue and in order to be able to develop effective
treatments.
Expression of violent behavior is influenced by a complex and dynamic interplay of biological, psychological
and social variables. Several studies have reported that Individual differences in aggressive behavior are at least
partly heritable and presumably result from the interaction between genetic and environmental factors. Geneenvironment interactions refer to genetic differences in susceptibility to particular environmental risk factors. Early
life environmental risk factors have detrimental effects on the long term mental health of individuals and increasing
evidence suggests that genotype can moderate the capacity of early environmental pathogens to alter the risk of
mental disorders. Several studies have suggested a relationship between MAOA polymorphism and aggression yet
the intervening neural mechanisms are still unclear. It has been suggested that individuals with the low expression
allele might be more sensitive to negative social experiences which might result in defensive aggressive behavior.
However, the link between genetics and behavior could be better understood studying brain response. In this sense,
neuropsychological, electrophysiological, functional and structural brain imaging parameters could represent the
genetic basis more directly.
The purpose of the present study is to investigate the relationship between the MAOA polymorphism, trait aggression,
early trauma, neuropsychological profile and neural organization (anatomy and function) by measuring structural
and functional MRI and the event related potentials in a group of 45 male healthy volunteers and 45 male subjects
with a behavioral history of aggression and violent behaviors.
Method
Behavioral Measures: Participants will complete several self-report measures related to impulsivity, aggression, early
trauma and hostility. All the measures have been standardized and validated in the Mexican population.
High resolution MRI images will be obtained in each subject. Also the Event Related Potentials and fMRI will
be recorded while observing pictures of emotionally charged scenes with and without moral content as well as
emotionally neutral pictures.
Neuropsychological measures will include a battery that assesses frontal lobe functions and several measures of
Attention and Memory.
Genotyping: DNA will be obtained with Orasure oral specimen collection device and extracted with Puregene DNA
purification kit. The MAOA-u VNTR polymorphism will be identified using polymerase chain reaction (PCR).
In sum the present study could help to clarify some of the psychological, affective and neurocognitive correlates
of the MAOA-aggression link and help to provide a better understanding of the experiences that mediate genebehavior relationships and eventually lead to better preventive and treatment alternatives.
*Contact
Phone: (55) 5622-2327
e-mail: [email protected]
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Other projects on health
Participation of cellular proteins and cholesterol
in dengue virus replication
Alcaraz Flores S1, Agis Juárez R1, Matos A1 and Del Angel RM1*
1Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados-Instituto
Politécnico Nacional, México, D.F.
Abstract
Dengue virus, a mosquito-borne virus, member of the Flaviviridae family causes dengue fever, dengue hemorrhagic
fever and dengue shock syndrome. The positive polarity RNA genome, contains a type I Cap in the 5’ end and lacks
of a poly(A) tail in the 3’ end. The open reading frame of the viral genome encodes for three structural and seven
nonstructural proteins. Flanking the open reading frame, the viral RNA contains two untranslated regions (UTR) that
are involved in various functions such as translation, replication and assembly The 3’-UTR contains a conserved
stem-loop (3’SL), two conserved sequences (5’- and 3’-CS1), and two complementary regions (5’UAR and 3’UAR)
that together induce dengue virus cyclization. Additionally, the UTRs from the positive and negative strands bind
cellular proteins such as La, polypyrimidine tract binding protein (PTB), the translation initiation factor eEF-1a, Y
box 1 protein (YB-1), poly A binding protein (PABP), protein disulfure isomerase (PDI) and calreticulin. One of the
objectives of the present project is to determine the role of these proteins in dengue virus replication. To analyze this
aspect, we will use a dengue virus replicon, where the Luciferase gene substituted the structural proteins. Silencing
of each of the cellular proteins described before, transfection of the replicon and determination of Luc activity will
be the strategy designed to determine the function of these proteins in viral replication. Additionally, It has been
described that the replicative complex of different flaviviruses contains the two hydrophobic viral proteins, NS2A
and NS4A, the abundant luminal protein, NS1, and NS5 and NS3 proteins. However it is not known which other
viral or cellular proteins could be present in the replication complexes. This aspect will also be evaluated in the
present project using by proteomic analysis from the proteins immunoprecipitated using anti-NS3, NS1 and NS5
antibodies. Finally, translation and replication of dengue virus occur in close association with cellular membranes.
Electron tomography (ET) shows dengue virus induces membrane structures to be part of one endoplasmic reticulum
(ER)-derived network. It has been suggested that these membranes are rich in cholesterol The third aspect that will
be analyzed in this project is to evaluate the importance of cholesterol in viral replication. It is being evaluated using
dengue virus stably transfected Vero cells with dengue virus replicon. The cells will be grown in the presence of
drugs that sequester cholesterol and the Luc activity will be determined after different times post-treatment.
Actually we have been silencing PTB, PDI and calreticulin. Silencing of the three proteins alters viral expression
suggesting that these proteins play a role during viral replicative cycle. On the other hand, we found that PTB
translocates from the nucleus to the cytoplasm during dengue virus infection in Vero cells. Finally, disruption of
lipid rafts inhibits dengue virus replication indicating that cholesterol and lipid rafts are important during viral
replication.
*Contact
Phone: (55) 5747-3800 ext. 5647
e-mail: [email protected]
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Other projects on health
Combined immunization protocols to induce systemic and mucosal
(oral/nasal) immune responses in a swine model
Alvarado G1, Ramírez C2, Zenteno R3, Lazo G2, Vega López MA2*
1Facultad de Estudios Superiores Cuautitlán, Universidad Nacional Autónoma de México, 2Departamento de
Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados-Instituto Politécnico
Nacional, 3Benemérita Universidad Autónoma de Puebla.
Abstract
In México, almost 23 million cases of acute respiratory infections were reported in 2008. Moreover, it is recognized
that up to 18% of deaths in children under the age of five are caused by respiratory infections. Paradoxically, many of
these diseases are actually preventable, but most vaccination protocols use the intramuscular route, which induces
very good systemic (i.e. serum) response, but is unable to protect mucosal surfaces. Since mucosal surfaces are not
protected, pathogens infect them and although the patient may show no illness, it may became a healthy carrier of
the pathogen, potentially infecting susceptible persons.
Unfortunately, mucosal immunization frequently induces tolerance to the antigen or needs toxins or high doses of
antigen to induce immunity. Therefore, our group developed several protocols of parenteral and local immunization
in order to induce mucosal immune response. Our first goal was to find a proper animal model to carry out these
studies. We selected the swine because of the similarities in size, behavior and physiology with human beings and
our previous experience studying its intestinal immune system. Also we developed a quantitative ELISA to accurate
measure IgG and IgA antibodies in serum and mucosal secretions.
In two experiments we immunized weaned pigs with ovoalbumin (OVA) twice, by different routes and measured
anti-OVA specific antibodies in serum, saliva and nasal secretion (NS) afterwards. In the first experiment we used
the subcutaneous (SC) and transcutaneous (TC) routes, showing that the SC route was the most effective inducing
serum and mucosal immune responses, although the latter was delayed and short lived. In a second experiment, in
order to increase the mucosal response, we applied a third dose of antigen by the intranasal (IN) route. We found
that the IN boost increased the level and length of serum IgA response in all protocols and in saliva in SC/SC and SC/
TC immunized animals, and in NS in SC/SC immunized pigs. Furthermore, the TC boost also worked well for IgA in
NS of SC/TC immunized animals.
These results show that the swine is an appropriate experimental model where it is possible to induce mucosal
immune response using soluble (poorly immunogenic) antigens when administered by combined (systemic/local)
routes. Knowing the type of immune cells activated in different tissue compartments, how they migrate and reach the
mucosal sites, it is possible to design more rational immunization protocols to induce protective oral and respiratory
mucosal immune responses.
Acknowledgments: This work was partially supported by Instituto de Ciencia y Tecnología del Distrito Federal,
Conacyt (Project 60941) and Cinvestav-IPN.
We are grateful to MVZ Manuel Flores, and personnel of Cinvestav’s animal house (UPEAL) for technical
assistance.
*Contact
Phone: (55) 5747-3344
e-mail: [email protected].
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Effectiveness of DNA vaccines against mumps and herpes simplex
viral infections in animal models
Barrón BL1*, Herrera E1, Salinas J1, Pérez Ishiwara G2, García F2, Zepeda P2, Carrero J3
1Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional (IPN),
2Escuela Nacional de Medicina y Homeopatía (ENMyH)-IPN, 3Instituto de Investigaciones Bibliográficas,
Universidad Nacional Autónoma de México.
Abstract
Introduction: Several viral infections have been successfully controlled by immunization, however until now there is
a lack of vaccines against many infections like herpes simplex virus (HSV) infections; and furthermore, some of the
approved vaccines against viral infections like mumps vaccines have shown important side effects, mainly involving
the central nervous system (CNS).
Objective: The aim of this research was to evaluate the effectiveness of two plasmid constructs, previously designed
in our laboratories. The pcDNAHNβη construct encodes a highly conserved region of the viral mumps HN protein,
which is considered the major viral antigenic protein. The pcDNAgHβε construct encodes the HSV-1 gH protein,
which is one of the main fusion proteins for viral entry into the cell.
Methodology: Both constructs, pcDNAHNβη and pcDNAgHβε were used to immunize mice and hamsters, respectively
by intradermal ear inoculation. A week after the 2 nd boost immunization, mice were challenged to induce the HSV1 cutaneous zosteriform infection or HSV-2 genital infection. Hamsters were challenged intranasally to induce the
systemic mumps virus infection. The effectiveness of the constructs were evaluated by comparison with the course of
the infection (daily weight, signs of infection, hispathological studies and viral isolation from several organs, titer of
systemic neutralizing antibodies and linfoproliferation assays) in mocked immunized animals and vector immunized
animals.
Results: The pcDNAgHβε immunization specifically protected all the animals from the HSV-1 zosteriform infection,
so no one of them showed CNS infection. Even more, the construct immunization was able to compensate the
immunosupression induced by the viral infection. For the HSV-2 genital infection, the construct immunization
protected only 60% of the animals. The pcDNAHNβη immunization was also able to induce an specific immune
response, which produced a delay in the course time of mumps viral infection, together with a reduction of the virus
concentration in different organs.
Conclusions: All these results showed that both constructs induced a specific immune response which protected
against the virus whose gene was cloned in it. However, more studies are necessary to improve the immune response,
especially in the mucosal system, either respiratory or genital, which are the natural entry site for many viruses to
initiate an infection.
ICyTDF 2008-2009 financial supported Project.
*Contact
Phone: (55) 5729-6000 ext. 62377
e-mail: [email protected]
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Identification of genes associated with criminal behavior: genome-wide
mapping the human genome with 500,000 SNPs
Berumen J1,2*, Ostrosky Solís F3, Vázquez Mena O1, Juárez Torres ME1, Romero C3, Borja K3, Díaz K3,
Salgado JC3
1Departamento de Medicina Experimental, 2Facultad de Medicina, 3Facultad de Psicología, Universidad
Nacional Autónoma de México, México, D.F.
Abstract
Violent behavior is increasingly common in our society and now it is considered a public health problem. Many
studies on adopted individuals at early age have shown clear evidence that genetic factors are associated with
criminality, and specific genes with potential risk for criminal and violent behavior have been identified. The aim
of this work is the search of novel genes associated with criminality. The general design included the study of
candidate genes and the analysis of the whole genome in a group of 150 convicted individuals compared with a
group of 150 controls. Both cases and controls were evaluated clinically and psychologically in terms to explore the
violent behavior. The genetic polymorphisms of six previously associated genes (MAOA, SLC6A3, SLC6A4, COMT,
AR, and TPH) are being explored in cases and controls. The genome-wide analysis with 500,000 single nucleotide
polymorphisms (SNPs) is being explored only in the cases and the data will be compared with a group of controls
previously explored. Preliminary results will be discussed. The findings of this work will help to understand the
genetic bases of criminal behavior in order to provide tools for the development of new treatment or for the design
of specific prescription according to genetic polymorphism, as well as for the application of preventive therapy or
specific corrective rehabilitation.
*Contact
Phone: (55) 2789-2000 ext. 1281
e-mail: [email protected]
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Analysis of polymorphisms in Mexican-mestizo postmenopausal
women with osteoporosis and osteoporotic fractures
Canto P1*, Rojano D1,2, Ibarra R1, Coronel PA1, Vergara LA1, Salas RM1, Coral VR1,3
1Centro Médico Nacional 20 de Noviembre, Instituto de Seguridad y Servicios Sociales de los Trabajadores
del Estado, 2Unidad de Medicina, Física y Rehabilitación Región Norte, Instituto Mexicano del Seguro Social,
3Escuela Superior de Medicina, Instituto Politécnico Nacional. México, D.F.
Abstract
Introduction: Osteoporosis is defined as a systemic skeletal disease characterized by low bone mass and
microarchitectural deterioration of bone tissue, with a consequent increase in bone fragility and susceptibility to
fracture. Although the diagnosis of the disease relies on the quantitative assessment of bone mineral density (BMD),
which is a major determinant of bone strength, the clinical significance of osteoporosis lies in the fractures that arise.
Heritability data show that genetic factors determine up to 80% of the variance in BMD, which is a major predictor of
osteoporotic fractures. Common genetics polymorphisms in several genes (OPG, LRP5, ESR1, IL-6, VDR y COL1A1)
have been proposed as candidates for influencing bone phenotypes at the population level.
Objetive: Analysis of polymorphisms in the OPG, LRP5, ESR1, IL-6, VDR y COL1A1 genes, potentially associated
with osteoporosis and/or osteoporotic fractures in postmenopausal women with Mexican-mestizo ethnic origin, who
live in the Mexico City.
Subjects and Methods: The study was approved by the Institute’s Human Research Committee. We will include 250
postmenopausal women with osteoporosis and 250 postmenopausal women without osteoporosis; besides, we will
study postmenopausal women with osteoporotic fracture. All patients must have a Mexican-Mestizo ethnic origin
and will assess by questionnaire.
The diagnosis of osteoporosis is for BMD at the lumbar spine (L2-4) and femoral neck using dual energy X-ray
absorptiometry (DEXA). The fractures will be identifying either by questionnaire, medical reports and radiographic
documentation.
We will isolate genomic DNA from blood leukocytes from all women. Osteoporosis will define according to the
WHO lineaments for Latin-American women. The genotyping for all polymorphisms will be carried out by realtime PCR allelic discrimination TaqMan assays. This is a case-control study. Differences in allele and genotype
frequencies between groups are assess by X2 tests.
Results: At the moment, we studied 100 postmenopausal women and the genomic DNA was obtained. Their age was
of 59±6.75 years and all women had a Mexican-mestizo ethnic origin. Besides, all of them fulfilled the questionnaire
of confounder factors.
Conclusion: This is a preliminary results of this study.
This work is support by the Instituto de Ciencia y Tecnología del D.F., México; Grant: PICDS08-34.
*Contact
Phone: (55) 5627-6900 ext. 21665
e-mail: [email protected]
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FrpB1 is a Helicobacter pylori protein involved in iron acquisition
when hemoglobina humana is used as iron source
Carrizo Chávez MA1, González López MA1, Velázquez Guadarrama N2,
Olivares Trejo J de J1*
1Posgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de México, 2Departamento de
Infectología, Hospital Infantil de México Federico Gómez, México, D.F.
Abstract
Helicobacter pylori (H. pylori) is an helical shaped Gram-negative bacterium that infects various areas of the stomach
and duodenum. Many cases of peptic ulcers, gastritis, duodenitis and perhaps some cancer are caused by H. pylori
infection. This bacterium has the capacity to survive in several environments in the human. As any pathogen require
high concentrations of iron to grow in the host. An efficient mechanism to acquire iron is to express membrane
proteins capable of binding hemoglobin (Hb), they are termed receptors. This proteins can bind Hb using the motifs
named FRAP y NPNL. Interestingly, has been observed that H. pylori grows in media supplemented with Hb as
sole iron source. By in silico analysis has been seen that this pathogen contains three putative proteins which bind
Hb: FrpB1, FrpB2 y FrpB3. Until now has been tested that FrpB2 protein binds Hb and when it is expressed uses
Hb as only iron source. Nevertheless the role of FrpB1 protein remains unknown. In the present work, by in silico
analysis and using ChuA amino acid sequences from E. coli as probe was identified FrpB1 protein (Q9ZKX4) in
the H. pylori proteome, (NCBI, Expasy). FrpB1 amino acid sequence was aligned with the following sequences: E.
coli – ChuA (Q7DB97), H. pylori - FrpB2 (Q9ZKT4), Y. pestis-HmuR (A9QYI4), P. gingivalis-HmuR (Q7MUG9) and
N. meningitidis - HmbR (B6DYJ0) using ClustalW program. Remarkably, we found that FrpB1 has FRAP and NPNL
motifs. The 3D structure comparison by PyMol (V 0.99) program showed us that these structures correspond to
membrane proteins. In addition, FRAP and NPNL motifs were identified in external structure while that TonB motif
was observed in the base of the structure. All these findings support the idea that FrpB1 is an Hb-bindig protein from
H. pylori involved in iron acquisition like FrpB2.
This work was financed by Instituto de Ciencia y Tecnología del Distrito Federal (ICyTDF).
*Contact
Phone: (55) 5488-6661 ext. 15309
e-mail: [email protected]
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Identification of molecular and serologic markers for diagnostic
of tuberculosis and latent tuberculosis infection
Castañón Arreola M1*, Dorantes Torres CV1, Vargas Romero F1, Carranza Salazar C2, Castillejos López M2
and Mendoza Hernández G3
1Postgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de México, 2Instituto Nacional de
Enfermedades Respiratorias, 3Facultad de Medicina, Universidad Nacional Autónoma de México.
Abstract
Tuberculosis is a wide prevalent disease that affects mainly developing countries, where the incidence of disease
counts for approximately 80% of new cases. On this countries the health care systems are unable to detect and
trait disease to reduce incidence and prevent transmission chains. This disease is a chronic contagious infection
that affects people of all ages and predominating in young adults. Two billon people are latently infected with
Mycobacterium tuberculosis and 5 to 10% of them will develop active tuberculosis.
The most well known form of tuberculosis control is BCG vaccination, for which a variable efficacy rate from 0% to
80% has been reported. Although the causes of the variation in the protection offered by BCG are diverse, like: (i)
genetic host susceptibility, (ii) a wide range of virulence among M. tuberculosis strains, (iii) over attenuation of BCG;
(iv) exposure to environmental mycobacteria (MOTT). It is known that MOTT take part in human immune response.
In animal model BCG vaccine was blocked or mask by the previous sensitization with MOTT (up to now it has been
identified about 117 species not considerate true pathogens).
The lack of accurate and rapid diagnostics for tuberculosis impedes management of patients and disease control.
Many patients are never diagnosed, and contribute to the astonishing number of yearly deaths from tuberculosis
worldwide. For populations from areas of high endemicity, differences in response to specific antigens used in
conventional diagnostic test are not established between TB patients and HHC and the risk of transmission from
undetected cases requires widespread access to diagnostic services and early detection.
M. tuberculosis has a complex multiplicity of antigens with a diverse chemical and immune reactive nature, the
understanding of specific antigens expressed during early infection, disease, latency or reactivation and their
immunological characterization are key for the development of new diagnosis test. For this reason this project proposes
develop more efficient diagnostics test specifics for M. tuberculosis based on identification of proteins without cross
reaction with antigens of currently used BCG vaccine or widely distributed nontuberculous mycobacteria (MOTT).
After strain selection, antigen specific antibody ELISA titers of IgG, IgA and IgM against cell extracts or culture
filtrate proteins of M. tuberculosis clinical isolates (selected according their prevalence) and BCG vaccine will be
determinates. Then comparative analysis of proteomes between clinical isolates and BCG vaccine, let us detect
proteins differentially recognized in the proteome of these strains by TB patients, HHC and community people.
Furthermore we define a protein set which can be useful to detect TB infection and exposition to environmental
mycobacteria. Once developed, clinical assessments of the molecular and serology tests in populations are necessary
to ensure their sensitivity and specificity.
*Contact
Phone: (55) 5488-6661 ext. 15304
e-mail: [email protected]
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Study of stability in humans using the stimulus-response technique
based on head movements
Cruz MC 1, Cardiel E1, Hernández PR1*
1 Departamento de Ingeniería Eléctrica, Centro de Investigación y de Estudios Avanzados- Instituto Politécnico
Nacional, México D.F.
Abstract
Fall is a major problem for adult persons. Fractures or corporal injuries in 30% of the people are related with falls,
which are produced by cumulative body disorders due to the age, in many cases. These changes decrease functional
performance in the body, and elderly may become more susceptible to fall under any stress condition. The studies
about risk of fall, face several problems like lack in diagnosing approaches, classification methods, and the multifactorial causes of falling. This paper proposes the development of a novel technique based on engineering topics
for stability assessment of systems, using control theory knowledge, applied to the movements of the head when
a subject is submitted to a stimulus-response process. The technique allows a measure of trends of equilibrium
through changes that occur in a 2nd order mathematical model, which represents the adjusted response. According
to the test, the resultant oscillatory movements, describes the 3D dynamics of the compensatory movements of the
head of the subject when visual stimuli are involved. The equipment used for recording the movements of the head
consists of three infrared cameras which processes the 3D signal, the acquisition system to monitor the head using
spherical reflective markers. Then, Ariel performance Analysis System is used as software for analysis. The stimulus
consists in open-closed-open eyes, during five-ten-five seconds respectively. The responses follow a second order
model for a step input, commonly used in typical control. In this way, it is possible a characterization of the system
performance in terms of maximum overshoot and temporal measures such a rise time and settling time. Dynamic
behavior of the second order model is described in terms of two variables: the damping factor ζ, which represents
the oscillation status of the system and the natural frequency ωn of the oscillation. The population sample was 35
subjects between 6 and 85 years old, divided by decades. Fitting of the models was performed by using an iterative
algorithm. This indicated a good description of the model respect to the real signal. All the resultant roots or poles of
the characteristic equation in the complex plane were located at the left of the imaginary axis. It means, according
with control theory criteria, all the responses were stable in relative sense. Moreover, it is observed differences
among decades of age, suggesting trends of evolution, ripeness and deterioration, considering stability assessment.
This technique is fast, simple and noninvasive, that overcomes cognitive difficulties when adult persons and children
are being studied. Prevention of falls through objective indicators is pursued.
*Contact
Phone: (55) 5747-3800 ext. 6201
e-mail: [email protected]
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Taxol synthesis by bacteria
Flores Bustamante ZR1, Rivera Orduña FN1, Marsch Moreno R1, Dendooven L1, Zavala Díaz de la Serna FJ1,
Flores Cotera LB1*
1Departamento de Biotecnología y Bioingeniería, Centro de Investigación y de Estudios Avanzados-Instituto
Politécnico Nacional, México, D.F.
Abstract
Cancer is an important health and socioeconomic problem all over the world. In México, a woman with a gynecologic
neoplasia dies every hour. Mexico City (DF), Jalisco, Nuevo León and Baja California are the states with the highest
incidence rates. Taxol is a potent and widely used antitumor agent. Originally Taxol production depended on extraction
from the bark of yew trees (genus, Taxus), yet the low yield and limited availability of those trees have stimulated the
development of several other alternative sources over the past 20 years. Total synthesis of Taxol has been achieved,
but the low yield makes the method impractical. Cell culture systems have been successfully developed for large
scale Taxol production, but require long incubation times (40 days). Despite intensive research and the development
of other alternatives, Taxol is still an expensive drug. Consequently, its use is still far too expensive for many people
in the world. In our laboratory, a group of 72 bacteria were isolated from the surface sterilized tissues of the Mexican
yew (T. globosa Schelechtendal), a rare tree species. Organic extracts prepared from cultures of every bacteria, were
screened for the presence of Taxol. Among them, the extracts of seven strains were consistently positive to Taxol
when analyzed by a monoclonal competitive inhibition enzyme immunoassay (CIEIA). Although Taxol titers were
low and ranged from 30 to 300 ng/L, searching within the bacterial genomes can result in identification, isolation
and cloning of the genes involved in the Taxol biosynthetic pathway. Further study of these genes may serve to
engineer other host bacteria for heterologous production of Taxol and develop a fermentation production method.
Bacteria have a number of advantages in relation to current production methods for Taxol: a) a fast growth in
simple culture media, b) they can be cultured at a large scale and a high cell density, c) they have a high tolerance
to shear stress and d) they have a small genome that is easy to manipulate. Taxol producing recombinant bacteria
may have a considerable potential for development of economical and environmentally compatible processes for
Taxol manufacture. The aims of this project are to i) obtain the complete genomic sequence of two Taxol producing
bacteria, ii) identify the genes or operon involved in the synthesis of Taxol in the genomic sequences iii) cloning in E.
coli the operon or genes involved in Taxol biosynthesis of one of those bacteria and iv) demonstrate Taxol synthesis
in a recombinant bacterium.
*Contact
Phone: (55) 5747-3800 ext. 4384
e-mail: [email protected]
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Electrochemical corrosion study of alloys used as medical implants
Genescá LJ1*,Galicia AG1, González Rodríguez CA 1,Tribolllet B2
1Facultad de Química, Universidad Nacional Autónoma de México, 2CNRS/UPR15, Physique des liquides et
electrochimie, Paris, France.
Abstract
Most metallic alloys are susceptible to corrosion damage. Titanium, cobalt and stainless steel are examples of metallic
alloys used as medical implants since many years ago. These alloys present corrosion though.
A corrosion phenomenon is present when metal atoms are removed from a structural element and pass to its
surroundings by a chemical or electrochemical reaction.
The main objective of this study is to get a medical implant more corrosion resistant than those exist today. In order to
overcome this objective, electrochemical techniques are used in aqueous solution to evaluate corrosion degradation
of metallic alloys used as medical implants in an environment similar to that of the blood. Additional information
from scanning electron microscopy was carried out to help to understand the degradation mechanism.
Preliminary results practiced in titanium alloys and stainless steel have shown a higher corrosion rate for titanium
alloys than stainless steel. To explain this experimental result, a surface analysis has been used. Corrosion product
layer for titanium alloys shows an amorphous layer while stainless steel has shown a compact layer. Thickness
corrosion products layer is thinner to stainless steel than that for titanium alloys though.
Thickness evaluation has been carried out by electrochemical impedance spectroscopy in the high frequency range
(30 kHz to 100 Hz). Independent of solution concentration it has been found a diffusion phenomena in medium
frequency range (100 Hz a 1 Hz). J. Black et al.; [1] explain that titanium alloys are not totally biocompatible with
the organism when this kind of materials are put them into the human body. According to Black, titanium alloys
degradation consists of ion liberation causing damage to the kidney.
On the other hand, stainless steel degradation also consists of ion liberation however; in this case, iron ion is not
harmful to the human body. We believe that electrochemical impedance technique revealed phenomena mass
transfer in both alloys in medium frequency range.
Taking into account the presence of chloride ion in the test solution, specimens were observed at the final of each
experience in the scanning electron microscopy in order to identify the corrosion type. This kind of analyses showed
a pitting corrosion in both cases which was more significant in the stainless steel. Next work will allow proposing
a kinetic model to explain corrosion degradation of titanium alloys and stainless steel when they are in side of the
human body.
*Contact
Phone: (55) 5622-5237
e-mail: [email protected]
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Genetic structure, migratory influences and the genome-wide
association Studies in the Mexican mestizo population
Gómez R1, Magana JJ1, Castañeda Montes F2, Majluf A3,Cortés Reynosa P1,
Leyva García N1, Solano I2, Revilla Monsalve MC4,Muñoz ML2*
1Departamento de Genética, Instituto Nacional de Rehabilitación, 2Departamento de Genética y Biología
Molecular, Centro de Investigación y de Estudios Avanzados-Instituto Politécnico Nacional, 3Unidad de
Investigación Médica en Trombosis, Hemostasia y Aterogénesis, Hospital General Regional Gabriel Mancera,
Instituto Mexicano del Seguro Social (IMSS), 4Centro Medico, IMSS, México, D.F.
Abstract
The Human Genome Project has stimulated interest in genetic determinants of diseases. The determinants of common
mendelian diseases that involve a single gene are well established. However, the current research addresses the
role of genetics in the complex diseases causing the major human morbidity and mortality, which result from the
concomitant effect of environmental, behavioral, and poligenetic factors.
Mexican mestizo population is a pluriethnic population, composed by a differential mixture of Native American,
and a recent migration from European and African. Genetic studies may yield misleading results in their disease and
non disease population that includes a different ethnic/racial mix; this particular form of confounding is referred to
as population stratification. Then, genome-wide association studies help to dissect the etiology of complex diseases.
Base on this, our major aim is to determine the genetic structure in a population of 500 unrelated individuals from
the central region of Mexico.
To develop this study, DNA was isolated using QIAmp DNA Blood Kit (Qiagen®). Polymerase chain reaction
amplification was done using AmpFlSTR Identifiler kit ®(Applied Biosystems) and the DNA typing was done using
capillary electrophoresis (ABI PRISM 310 Genetic Analyzer, Applied Biosystems). According to the analysis of 15
STR loci by the Structure software (Pritchard, et al 2000) the population of Mexico City is genetically substructured
in at least 4 subpopulations (p=0.99985178). In conclusion, genome-wide studies should consider this admixture
like a correction statistical factor to avoid spurious disease-marker association or mask a true association. We also
find evidence that the mixture at the origin of these populations involved mainly Native, African and European
immigrants. Our results also suggest that larger urban agglomerations like Mexico City attracted immigrants from
relatively distant areas, thus potentially tracing their ancestry to various differentiated Native groups. This patern
agrees with demographic data showing that expansion of this city has been driven by regional immigration rather
than by internal growth.
This work was supported by grant from the Instituto de Ciencia y Tecnología del Distrito Federal (ICyTDF-MLMM).
*Contact
Phone: (55) 5747- 3800 ext. 3335
e-mail: [email protected]
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Genomics, proteomics and bioinformatics: An integral approach to the study
of metabolic regulation in the yeast Saccharomyces cerevisiae
González A1*, Hernández H1, Herrera J1, Aranda C1 and Del Río G1
1Instituto de Fisiología Celular, Universidad Nacional Autónoma de México. México, D.F.
Abstract
In Saccharomyces cerevisiae, transcription of genes participating in biosynthetic pathways is elicited during amino
acid deprivation through the GCN4-encoded transcriptional activator. Conversely, when this yeast is provided with
secondary nitrogen sources, transcription of genes coding for enzymes involved in their catabolism is determined
through the action of Gln3p, which constitutes the main activator of the Nitrogen Catabolite Repression network.
Thus the role of each one of these activators has been secluded to either biosynthetic or catabolic pathways and
no synergic interaction between then has been considered. This study addresses the question of whether Gln3p
and Gcn4p cooperatively determine expression of their target genes. To this end we have analyzed expression of
four catabolic and two biosynthetic genes under nitrogen repressive or derepressive conditions in yeast cells grown
with or without amino acid deprivation. Results show that Gln3p and Gcn4p form part of a complex determining
transcriptional activation of catabolic and biosynthetic genes when cells are grown on a secondary nitrogen source
with or without amino acid deprivation.
The Hap complex, is constituted by four subunits, the HAP2, HAP3, HAP5 encoded polypeptides constitute the
DNA binding domain, while the HAP4-encoded protein constitutes the activation domain. Although it has been
determined that this complex fosters transcriptional activation of genes involved in respiratory metabolism, our
results suggest that it can form a peculiar complex which includes Gln3p, and participates in the regulation of genes
pertaining to the Nitrogen Control Circuit.
The general objective of this work is to analyze molecular interactions between Gcn4p and Gln3p and the Hap
complex and Gln3p, in order to provide evidence indicating that transcriptional activators can interact to give
rise to new complexes which determine a peculiar transcriptional response. We have analyzed transcriptional
regulation of the pertinent set of genes, and single and double mutants differentially tagged in the pertinent genes
have been constructed in order to carry out chromatin immunoprecipitation (Chip) and coimmunoprecipitation
experiments. To determine which is the set of genes whose expression is regulated by the Gcn4p-Gln3p and HAPGln3p transcriptional complexes, microarray experiments are currently being carried out.
Considering that the number of applications that have been developed to analyze microarray has enormously
diversified, we are developing a server that will furnish a microarray analysis service that will be offered to those
groups interested in analyzing microarry generated information through the combination of various computational
approaches. The server will offer a compendium of tools combining Internet Search methodologies (WormWeb),
text mining and clustering (Self-Organized Maps), this facility will constitute part of the services offered through the
creation of a company and a license of the new methodology which is being developed.
*Contact
Phone: (55) 5622-5631 ext. 25631
e-mail: [email protected]
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Human caliciviruses detected in Mexican children admitted to three hospitals
from Mexico City with severe acute gastroenteritis
Gutiérrez Escolano AL1*, Velázquez R2, Escobar Herrera J1,
López Saucedo3, Torres J2, Estrada García T3
1Departamento de Infectómica y Patogénesis Molecular, 3Departamento de Biomedicina Molecular, Centro de
Investigación y de Estudios Avanzados-Instituto Politécnico Nacional, 2 Hospital de Pediatría, Centro Médico
Nacional SXXI, Instituto Mexicano del Seguro Social, México D.F.
Abstract
Gastroenteritis is a major cause of morbidity and mortality worldwide, resulting in at least 2.5 million deaths per
year in children under 5 years of age in developing countries alone. This disease can be caused by a number of
organisms, including many viruses and bacterial pathogens; however, viruses pose a particular problem owing to
the limited number of antiviral therapies available for treatment. Over the past decade, it has become evident that
Human caliciviruses (HuCVs), members of the Norovirus (NoV) and Sapovirus (SaV) genera of the Caliciviridae
family, are the major cause of outbreaks of non bacterial gastroenteritis worldwide. The frequency with which these
viruses affect closed and semi-closed communities, such as hospitals, nurseries, military groups, hotels and touristic
cruise, has revealed its impact on public health. Only in the United States, 23 million cases occur annually, however,
despite the economic impact and considerable morbidity, there is no medication or vaccines to treat or prevent this
disease.
Due to severe characteristics of the life style in Mexico City, such as the high consumption of food from street
vendors, the high population density, a deficient sewer system, and general poor hygienic conditions, our population
is in increased risk to acquire gastrointestinal diseases.
In this regard, the presence of HuCVs infection in Mexico has been documented. A high prevalence of serum
antibody and the co-circulation of multiple clusters of symptomatic and asymptomatic NoVs and SaVs were reported
in children from a peri-urban area of Mexico City. Furthermore, NoV infections have been identified as a cause of
traveler’s diarrhea. Nevertheless, little is known about the participation of these viruses as a single cause of acute
diarrhoea episodes in children requiring hospitalization.
In the present study, a total of 1129 children ≤5 years old, hospitalized due to acute diarrhea, were enrolled during
March 1998 to December 2000, from 3 main hospitals in the northern (Tlatelolco Hospital), southern (Villa Coapa
hospital), and center (Hospital Gabriel Mancera) areas of Mexico City. After analyzing all faecal samples for several
enteropathogens (Salmonella spp., Shigela spp, Vibrio cholerae, Campylobacter spp., Aeromonas spp. diarreogenic
E. coli pathotypes, Giardia lamblia, Cryptosporidium spp., Cyclospora spp., Isospora spp., Entamoeba hystolitica),
396 stools that remained negative, were further screened for HuCVs by RT-PCR. HuCVs were detected in 22/396
samples; 19 belong to NoV GII and one to SaV GI/2. Associations between the presence of HuCVs and clinical and
epidemiological data revealed that, NoV associated diarrhea, occurred with a seasonal pattern, and that children
hospitalized due to human calicivirus disease scored an average of 13 points on the Vesikari scale, which corresponds
to severe episodes. These results highlight that human caliciviruses by themselves are etiological agents of acute
severe diarrhoea among children that required hospitalization, and that its detection is important in order to reduce
the diagnosis gap in Mexican children.
*Contact
Phone: (55) 5747-3341
e-mail: [email protected]
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Development of physiotherapy ultrasound equipment
Gutiérrez MI1, Vera A and Leija L1*
1Departamento de Ingeniería Eléctrica, Centro de Investigación y de Estudios Avanzados-Instituto Politécnico
Nacional, México, D.F.
Abstract
Medical Ultrasound is commonly associated with applications for diagnosis which use low intensity pulsed
ultrasound (less than 0.1 W/cm2) to examine soft tissues. Nowadays, many research groups are researching about
new therapeutic alternatives related with the applications of higher intensity ultrasound. Ultrasound therapy does not
just refer to a specific therapy, but to the use of ultrasonic energy to induce changes in the tissue condition through
out thermal and mechanical effects. The data of therapeutic effects of half intensity ultrasound (between 0.1 W/cm2
to 3 W/cm2) have been gathered from different parts of the body like brain, kidney, liver, bone, muscle and some
ocular tissues, but the way of how this energy interacts with tissues is not yet clear. Research about the topic has
not yet finished and it requires specific work in the factors that change the behavior of ultrasonic energy in different
media.
In this poster, it is proposed the construction of an ultrasonic equipment to generate a continuous or pulsated
ultrasonic wave to be used in research, and later on in hospitals. At first, the equipment was designed based on some
characteristics, as high frequency stability, high power stability, adjustable frequency and power in accordance with
the transducer, self-protection systems and high efficiency of energy conversion. The device was used to investigate
the phenomena involved in energy conversion, i.e. the conversion of ultrasound into heat. Temperature distributions
were gotten for analyzing them through computational modeling, in order to determine the interaction of this energy
with tissues. At the end of the research, it is expected to have a new protocol for physiotherapy and a new system
with technological innovations.
The developed device for ultrasonic physiotherapy works at a variable frequency from 700 kHz to 1.6 MHz in
increments of 1 kHz. Its power can be controlled analogically from 0 to 2 W/cm2 and the duty cycle can be easily
adjusted. The employed transducer is a commercial one and it works at 2 W/cm2 maximum. A system was added to
determine the lack of load and to avoid ultrasonic emission to air which may damage the transducer. The complete
equipment is composed of a frequency synthesizer, an RF amplifier, a control system (of frequency and power), a
load-detection system, and a specific power supply. The device was tested in a hospital, and now it is being modified
to get an innovatory system.
It is known that Mexico has a technological gap due to, among other things, the acquisition of the most of equipments
in foreign companies; few of these ultrasound equipments are developed here in Mexico. Technological development
can be improved when our own devices are designed by ourselves. If we are the authors of our technology, we can
improve it to get either just a better device or even to compete in the international market. With this kind of actions,
the technological dependence would be stopped (or at least decreased) and, in consequence, the medical services
would be enhanced. This ultrasound equipment can be manufactured at low cost and used in DF’s hospitals.
*Contact
Phone: (55) 5061-3850 ext. 3850, 5202
e-mail: [email protected]
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Development of non-invasive biomarkers for the diagnosis of fibrosis
Kershenobich D1*, Guzman C1, Kershenovich R1, Zlotnik A2, Dehesa M3, Selman M4, Gutierrez Reyes G1
1Departamento de Medicina Experimental, Universidad Nacional Autónoma de México, 2University of Irvine
California, 3Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, 4Instituto Nacional de
Enfermedades Respiratorias, México, D.F.
Abstract
Introduction: Fibrosis is a response to different etiologic agents in which scar tissue potentially affects various organs
including, liver, pancreas, lungs, heart, kidneys, etc., this being a very important problem regarding morbidity and
mortality.
A common characteristic in fibrosis is that the lesioned regions independently of which organ is affected are
encapsulated by an extracellular matrix composed by collagen, glycoproteins, and proteoglycans.
The progression of fibrosis is a dynamic process in which stellate cells have been identified as the most important
cellular element.
Several studies have demonstrated that fibrosis can be reversible, however, the exact moment in which this happens
is unknown.
The clinical importance of monitoring fibrosis radicates basically in the next aspects:
1.- The morbimortality of many chronic degenerative illnesses are related to the state and progression of fibrosis.
2.- Stabilization and regression have been demonstrated in the context of the specific treatment of the basic
pathology
3.- The presence of fibrosis in some situations is an indicator to initiate or terminate treatment.
4.- The actual goal standard for diagnosing and monitoring fibrosis is the biopsy, which has many limitations: invasive
procedure, expensive, inter-observer dependant. Because of this we are in need of finding non-invasive imaging and
serologic methods for the knowledge of fibrosis status in a patient. The limitations of the non-invasive studies we
have today are the impossibility to differentiate moderate from severe states, the impossibility of classifying 40-60%
of the patients and the low sensibility in relation to the effects of treatment. The main practical and scientific problem
by which we have decided to make this research proposal is the need to develop and validate a non invasive
method, both for diagnosing and follow up of fibrosis in chronic degenerative illnesses.
By this we will estimate more efficiently, the status of the pathological process, its natural history and be able to have
a valuable indicator of progression and a key for monitoring the response to antifibrotic molecules.
Fibrosis is a very relevant pathology in medicine and public health offering the opportunity to develop new
technologies for diagnosis and/or treatment.
Objectives:
1. Validation of a panel of serologic biomarkers that reflect pathophysiology of the fibrogenic process in
chronic degenerative illnesses.
2. Demonstrate the biologic activity of these biomarkers.
3. Determine the reproductibility and differentiation of this panel in different fibrotic stages.
4. Automatic construction of decision trees for the classification of fibrosis.
Study: We will study 100 patients, 50% with Hepatitis C and 50% with idiopathic pulmonary fibrosis; each patient
will undergo a complete medical evaluation including blood tests, imaging studies, and tissue biopsy.
We will study a number of specific proteins expressed in the liver in relation with fibrosis and their presence in
different state of the disease previously identified and compare them to the methods employed at the present time.
Preliminary results: At this moment we have recluted 8 candidates for the study.
*Contact
Phone: (55) 5623-2673
e-mail: [email protected]
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Proteomic biomarkers of osteoarthritis in early stages diagnosis
Kourí Flores JB1*
1Centro de Investigación y de Estudios Avanzados-Instituto Politécnico Nacional, México, D.F.
Abstract
Osteoarthritis (OA), a musculoskeletal disorder included in the Bone and Joint Decade, is the most common form of
arthritis affecting millions of people worldwide and the major cause of disability in the elderly. Significant proportion
of population above the age of 45 years is affected by this nonfatal, but crippling disease. Its prevalence is more
in women than men. OA and its other related conditions increase dependency of elderly on others, and result in
enormous losses in medication, and later surgery, thereby affecting Nation’s economy. Various invasive and noninvasive methods are reported for the diagnosis of this articular cartilage pathology. (X-ray, computed tomography,
magnetic resonance imaging, arthroscopy and arthroscopy) are having their disadvantages, however the diagnosis of
OA in early stages with simple effective noninvasive methods are still missing. Chondrocytes, which are the unique
cellular component of adult articular cartilage, are capable of responding to structural changes in the surrounding
cartilage matrix. Since the initial stages of OA involve matrix mechanical disruption before the inflammatory process
appears, it will be important to find matrix molecules as targets for therapeutic intervention markers before the
inflammation process starts. Current pharmacological interventions that address chronic pain are insufficient and
no proven disease-modifying therapy is available. Identification of methods for early diagnosis is of key importance,
since therapeutic interventions aimed at blocking or reversing structural damage will be more effective when there
is the possibility of preserving normal homeostasis.
In OA, the most interesting aspect of cartilage proteomics is the ability to compare the composition of cartilage
protein profiles coming from healthy and early osteoarthritic joints and to identify proteins of which the concentration
and/or the nature of modifications differ. At present, the diagnosis of OA primarily rests on the appearance of pain,
when the deterioration of cartilage has already reached a point of no return, and patients are generally treated
symptomatically with anti-inflammatory drugs or analgesics.
Our group is interested in developing appropriate technical skills to provide early detection of the disease to prevent
and / or stop the development of damage in patients’ joints.
*Contact
Phone: (55) 5747-3800 ext. 3343
e-mail: [email protected]
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Identification, characterization and IgE recognition of profilin
from Amaranthus palmeri pollen
Landa Pineda CM1, Rosas Alvarado A2, Benítez Cardoza CG1, Zamorano Carrillo A1, Cabrera Avila AL1, Terán
LM3, Mendoza G4, Reyes López CA1, Marchat LA1*
1Posgrado en Biomedicina Molecular, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico
Nacional, 2Laboratorio de Inmunología y Alergia, Hospital General de México, 3Departamento de Investigación
en Inmunogenética y Alergia, Instituto Nacional de Enfermedades Respiratorias, 4Facultad de Medicina,
Universidad Nacional Autónoma de México. México, D.F.
Abstract
Immunoglobulin E-mediated allergies are the most prevalent of human disorders, affecting almost 30% of the
population in developed and developing countries. The symptoms of allergy (e.g., rhinoconjunctivitis, asthma,
food allergy, dermatitis, and anaphylactic shock) are caused by IgE recognition of per se harmless allergens. In
allergic patients, allergen contact causes the immediate release of inflammatory mediators from mast cells, as well
as chronic T cell- and eosinophil-mediated tissue inflammation. Allergic tissue inflammation can be mitigated using
anti-inflammatory drugs and immunosuppressive agents. However, only allergen-specific immunotherapy, the
administration of gradual increasing quantities of the disease-eliciting allergens, may be considered as causative
treatment. Allergen-specific immunotherapy has long-lasting clinical effects and may prevent the progression of mild
forms of allergy to severe manifestations. To reduce allergenic side effects, several immunotherapy studies have been
conducted using T cell epitope containing peptides with no or low IgE reactivity. More recently, genetically modified
hypoallergenic recombinant allergen derivatives have been engineered. Detailed molecular and immunological
studies of the major allergens are indispensables to the identification and design of hypoallergenic molecules to be
used in the allergen-specific immunotherapy.
Recent reports of scientific societies of allergy indicate that around one half of allergic people present reactions to
pollen allergens. Furthermore, pollen-allergic patients frequently present allergic symptoms after ingestion of several
kinds of plant-derived foods. Pollen allergens offer a dual perspective of study: some of them are considered key
proteins for pollen physiology, but they are also able to trigger allergy symptoms in susceptible humans after coming
in contact with their tissues.
Profilin is a relevant allergen present in pollen and fruit and this is responsible of part of cross-reactions between
different allergens sources. This allergenic protein is an actin binding protein of low molecular weight (12-16 kDa)
that is present in all eukaryotic cells, from yeast to man. Pollen from Amaranthus palmeri is one of the main allergens
sources in the Metropolitan zone of Mexico City, and clinical studies showed that this is a relevant allergen source.
To date, there are no detailed molecular studies of clinically relevant allergens from this pollen and their biological
functions, as well as their cross-reactions with other pollen allergens. In this work, we detected and isolated four
profilin isoforms from A. palmeri pollen and we present the results obtained of the IgE recognition from sera of
allergic people to this allergen and their clinical relevance in the sensitivization of allergic Mexican people.
*Contact
Phone: (55) 5729-6000 ext. 55562,
e-mail: [email protected], [email protected]
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The clinical DNA diagnosis laboratory of UACM: a new scientific tool
to face critical problems of Mexican population
López Casamichana M1
1Posgrado de Ciencias Genómicas, Universidad Autónoma de la Ciudad de México. México, D.F.
Abstract
Genetic information understanding of living organisms has revolutionized the biological and medical sciences.
The possibility of looking inside and manipulating genomes implies technological, economical, social, political,
religious and scientific repercussions. In medicine, anthropology and forensic issues, the approaches include, for
example, target sequence identification, findings of mutations and polymorphisms on DNA, as well as genotyping
and confirmation of deletions and insertions in cell genomes. Mexico City epidemiological scenery shows that
HIV and cancer are moving on, tuberculosis and influenza are reappearing, and parasitic infections continue
being a problem among the marginal population. Simultaneously, the incidence of neurodegenerative and chronic
degenerative diseases has increased in oldest population. These facts reveal that there still are several failures in
Mexican health system which require special attention and impose new challenges as well.
In respond to these defiances, the “Universidad Autónoma de la Ciudad de México (UACM)” and the “Instituto de
Ciencia y Tecnología” of Mexican Federal District (ICyT-DF) celebrated an agreement to establish commitments in
order to maximize human, material and financial resources by developing a Clinical DNA Diagnosis Laboratory. So
far, we have performed actions involving strategic designing and construction of specialized areas and the acquisition
of technological equipment. That means we have invested 2,5 million Mexican pesos in specialized equipment for
DNA extraction, quantification and molecular analysis and we have designed and created a safety and advanced
laboratory for molecular diagnoses through clinical DNA testing.
Our main goal in the near future is to implement front line scientific methods in molecular detection, designed for
sorting and monitoring of pathogen agents in clinical samples. Moreover, we are planning to make specific analysis
to diagnose human genetic disorders which induce several affections, such as neurodegenerative, cardiovascular,
metabolic and cancer diseases.
Another interesting point to deal with is the Genetic fingerprinting or DNA profiling. These molecular analyses
facilitate human identification precisely and hold a high impact in justice and security administration. In this respect,
we shall collaborate with government institutions providing a scientific approach.
As a tighter link between genomic sciences and society becomes more necessary as time passes by, the existence
of the Clinical DNA Diagnosis Laboratory of UACM will confirm its relevance to face critical matters of Mexican
population.
*Contact
Phone: (55) 5850-1901 ext. 15301, 15305
e-mail: [email protected]
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Genomics of myotonic dystrophy type 1 (DM1): molecular diagnosis
and identification of genes with therapeutical potencial for DM1
Magaña JJ1, Cortés Reynosa P1, Rodríguez R3, Gómez R1, Escobar RE2, Leyva N1, Cisneros B3*
1Departmento de Genética, 2Departmento de Electrofisiología, Instituto Nacional de Rehabilitación;
3Departmento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados -Instituto
Politécnico Nacional, México, D.F.
Abstract
Myotonic dystrophy (DM1) is a multisystem disorder considered the most common form of muscular dystrophy in
adults with an incidence of 1:8000: DM1 is caused by the expansion of a trinucleotide CTG repeat in the 3’-untranslated
region (3’-UTR) of the DMPK gene, localized in the chromosome 19q.13.3. Pathologic alleles contain more than
50 trinucleotides and can expand up to several thousand repeats. Extensive studies have been performed to explain
how an untranslated mutation can lead to a dominant pathogenic phenotype. Currently, the most accepted model
for DM1 proposes a gain-of-function RNA mechanism, in which the mutant mRNA is trapped in the nucleus altering
the gene expression and alternative splicing of various genes. The aim of this study is to establish a cost-effective
molecular diagnostic test for DM1 for its clinical use in the National Rehabilitation Institute, a public reference center
for DM1 that offers service to the uninsured population in our City. Our second goal is to identify genes altered by
the DM1 mutation in muscular tissue of DM1 patients by means of a genome wide microarrays study, as a first step
to develop gene therapy strategies for the disease. To accomplish the first objective, we analyzed the DM1 locus in
400 unrelated healthy subjects of the Mexican mestizo population. We found twenty five different, ranging from 5 to
37 repeats, The distribution of alleles was bimodal, with peaks at 5 and 10-14 repeats and being the most common
alleles those containing 13, 11 and 5 CTG repeats. Polymorphic distribution of our country was compared with 8
different human populations (Caucasian European, Yugoslav, Japanese, Korean, Taiwan, African Negroids, African
American and Chilean) and one Mesoamerican group (Mixtec), finding a high influence of the Native American
groups in our population.
To accomplish our second goal, we have been developed skeletal muscle cell cultures from DM1 patients and
healthy subjects by differentiating fibroblast cells with overexpression of MyoD, a “master” gene that controls muscle
differentiation. Such strategy avoids the invasive procedure of taking muscle biopsy from patients, and will enable
us to perform the microarrays studies.
*Contact
Phone: (55) 5255-5747 ext. 3339
e-mail: [email protected]
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Evaluation of immunogenicity and protective efficacy
of a pcDNA-Ehcpadh vaccine in hamster
Martínez MB1, Rodríguez MA2*, García Rivera G2, Sánchez T2, Hernández Pando R3, Aguilar D3, Orozco E2
1Posgrado en Ciencias Genómicas. Universidad Autónoma de la Ciudad de México. 2Departamento de
Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados-Instituto Politécnico
Nacional, 3Departamento de Patología, Instituto Nacional de Ciencias Médicas y Nutrición, Salvador Zubirán,
México D. F.
Abstract
Entamoeba histolytica is a protozoan parasite, with a worldwide distribution and responsible agent of human
amoebiasis disease. Throughout the world, 40 million of individuals are affected by amoebiasis and around 100 000
of them die due to this parasitic disease every year. EhCPADH is an immunogenic and heterodimeric protein of E.
histolytica formed by a cysteine protease (EhCP112) and an adhesin (EhADH112). This protein complex is located
in the plasma membrane and in cytoplasmic vesicles of trophozoites, and several studies have been demonstrated
that it is involved in cytopathic effect, target-cell adherence and phagocytosis. In previous reports, we showed that
EhCPADH could be a good vaccine candidate to prevent invasive amoebiasis. In this study, we immunized hamsters
with a pcDNA-Ehcp112/pcDNAEhadh112 plasmid mixture (pcDNA-Ehcpadh) by intramuscular or intradermal route.
Then, we evaluated in the immunized animals: i) the expression of the amoebic genes; ii) the humoral response; iii)
the cell mediated immune response; iv) the cytokine pattern associated to the cellular immune response before and
after an intraportal inoculation of virulent trophozoites; and v) the hepatic damage and survival rate of challenged
animals. RT-PCR and immunohistochemical assays showed that both antigens were expressed in spleen and liver of
immunized animals, but they displayed different expression patterns. No antibodies immune response was obtained
for none of the two routes. However, intradermally immunized hamsters showed a robust Th1-like immune response,
characterized by high levels of INF- and TNF- cytokines, detected in the liver of hamsters challenged with virulent
trophozoites. Vaccination of animals by the intradermal route results in a considerable reduction of hepatic abscesses
formation and in a higher survival rate after the challenge with virulent trophozoites. These results suggest that a
refinement of this DNA vaccine could be a good choice to control hepatic amoebiasis.
*Contact
Phone: (55) 5747-3800 ext. 5653
e-mail: [email protected]
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A dendritic cell precursor in peripheral blood correlated
with psoriasis activity
Mora Velandia LM1, Pérez Montesinos G1, Isibasi A2, Santa Cruz FJ3, Bonifaz LC1*
1Unidad de Investigación Médica en Enfermedades Autoinmunes, 2Inmunohistoquimica, Hospital de
Especialidades, Centro Médico SXXI, Instituto Mexicano del Seguro Social, 3Centro Dermatológico Ladislao de la
Pascua, Secretaria de Salud, México, D.F.
Abstract
Dendritic cells (DC) play a pivotal role in the development of immune responses and the maintenance of peripheral
tolerance. Psoriasis is one of the common cutaneous autoimmune diseases in Mexico City. Psoriasis diagnosis
and prognosis is mostly clinical supported with skin biopsy. Specialized DC subsets in skin have been reported
to participate as key orchestrators in the pathogenesis of psoriasis. However, the contribution of dendritic cells or
their precursors from the periphery in the pathogenesis of skin lesions is unknown. Conventional dendritic cells are
characterized by the expression of CD11c, CD11b and MHC-II. In this work we evaluated DC subpopulations in
peripheral blood of psoriasis patients with different grades of the disease and compare them with normal donors
(n=20) and found in patients with high disease activity a dramatic reduction of a population that express low levels
of CD11c and MHC-II but is positive for CD11b and CD123 (IL-3R). This population also expresses high levels of the
cutaneous antigen (CLA). The identified population freshly isolated from peripheral blood does not have dendritic
cell morphology or phenotype. However after culture with IL-3 acquires a DC phenotype expressing CD11c and
MHC-II. The loss of this population in peripheral blood correlates with both psoriasis activity and evolution time
of the disease (R=0.827) suggesting that the identified population could be a DC precursor able to migrate into the
skin to participate in the pathogenesis of psoriatic lesions. The inverse correlation of the presence of this population
in peripheral blood with psoriasis activity can be used as a blood marker for the prognosis of the disease. The
identification of this population and its function in psoriasis and other prevalent cutaneous diseases in Mexico City
can be also important to design new therapeutic tools.
*Contact
Phone: (55) 56276900 ext. 21370
e-mail: [email protected]
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Molecular markers for the identification of resistance to antifungal
(itraconazol and voriconazole) in clinical isolates of Aspergillus fumigates
Reyes Montes MR1*, Frías De León MG1, Duarte Escalante E1, Zavala Ramírez M1, Córdoba S2, Davel G2,
Bobadilla del Valle M3
1Facultad de Medicina, Universidad Nacional Autónoma de México, 2Departamento de Micología, Instituto
Nacional de Enfermedades Infecciosas (ANEI), Administración Nacional de Laboratorios e Institutos de Salud
(ANLIS) “Dr. Carlos G. Malbrán”, Buenos Aires, Argentina, 3Departamento de Infectología, Instituto Nacional de
Ciencias Medicas y Nutrición Salvador Zubirán, México, D.F.
Abstract
Aspergillus fumigatus is the causal agent of the disease aspergillosis, commonly distributed worldwide. In the last few
years, the association of the fungus with immunosuppressed hosts has increased considerably, as shown by diverse
authors. The treatment of these patients has been limited to amphotericin B, the wide spectrum triazoles, such as
itraconazol or voriconazole and/or echinocandin caspofungin. The continuous use of these treatments may result in
the development of resistance, therefore the use of these antifungal results minimally effective, with a mortality rate
between 30% and 90%. Unfortunately, in Mexico (MX), there is hardly any information available on the number
of cases, the sources of infection and their relation with the clinical forms caused by these fungi. In our country, A.
fumigatus is an important pathogen since the number of cases in immunosuppressed patients has increased from
there the importance of administering an effective treatment based on the selection of an adequate antifungal for
which this study´s purpose is to obtain SCAR markers (Sequence Characterized Amplified Region) that identify
resistance to itraconazol and voriconazole in A. fumigatus clinical isolates from MX.
From phenotypically characterized A. fumigatus isolates (macro and micromorphology, thermotolerance at 28, 37
and 48ºC, resistance profile to itraconazol and voriconazole), genetically homogeneous populations from sensitive
and resistant isolates to the antifungals will be selected from polymorphic patterns generated by AFLP with eight
combinations of oligonucleotides to identify the transcripts of differential expressions related to resistance for later
designing oligonucleotides based on the sequence of those transcribed. The SCAR markers are assessed in all of the
previously typified isolates as sensitive or resistant to both antifungals. To date, the phenotypical characteristics of 70
isolates of the fungi from MX, Argentina (AR), Peru (PE) and France (FR) have been analyzed. No variability was seen
in the macromorphology, however, the micromophology and speed of growth to the three temperatures studied,
we were found that the isolates from Peru grew at a faster rate and have larger vesicles with respect to the isolates
from MX, AR and FR. The PE isolates show specific phenotypical characteristics that differ them from the rest of the
isolates. Having SCAR markers for antifungal resistance for A. fumigatus, from autochthonous isolates from MX is
important as it allows for faster and specific diagnosis of the disease, as well as for identifying the adequate antifungal
for each patient using an accessible and easily transferable methodology for Mexican diagnostic centers and those of
other countries. Once SCAR markers are found, it is expected that an inexpensive kit will be marketed for identifying
the resistance to these antifungal.
*Contact
Phone: (55) 5695-9501
e-mail: [email protected]
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Development of mobile technologies, committed to health care,
beyond telemedicine
Salmerón Quiroz BB1*, Cano Arrieta G1, Guerrero Castellanos F1, Olmos Navarrete L1, Ruiz García JM1, Ruíz De
Los Santos MA1, Pérez Pedraza A1, Orozco Manzo PL1, Villegas Medina G1, Rodríguez Paredes SA1
1Escuela Superior de Ingeniería Mecánica y Eléctrica, Instituto Politécnico Nacional, México, D.F.
Abstract
The proposed project will contemplate the functional requirements and technological feasibility of integrating
computer systems on a moving ambulance with computer systems of a hospital and/or integrate these systems of
medical measurement in a mobile platform (wireless or data store in pen drives) useful to monitoring and diagnosis
of patients.
Therefore, in this project is pretended to create portable system, wireless for monitoring and diagnosis patients that
counts with the principal parameters of diagnosis, to be:
• Blood Pressure
• Heart Rhythm
• Respiration Rate
• Temperature
• Glucose Level
• Oxygen Saturation
• Body Mass Index
Such measurement system is wanted to be autonomous, portable, making the integration of mobile high speeded
low cost and low priced technologies.
Result: Feasibility study, Study and analysis of sensors to be deployed, Tracking trajectory algorithms development,
also used in tracking of indeformable objects, Real time data fusion software development, Human resources
configuration, Temperature sensor development, Pressure sensor development.
*Contact
Phone: (55) 5729-6000 ext. 64528, 64529
e-mail: [email protected]
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Production and purification of plasmid DNA vaccines
Sánchez Casco M1, Islas Lugo Fabiola1, Montes Horcasitas M. del C.1, Arroyo Rossana2, Martínez Benitez MB3,
Montesinos Cisneros RM4, Tejeda Mansir A4, Guzmán Zamudio R5, Aguilar Setién JA6, Dumonteil E7 and Ortega
López J1*
1Departamento de Biotecnología y Bioingeniería, 2Departamento de Infectómica y Patogénesis Molecular,
Centro de Investigación y de Estudios Avanzados-Instituto Politécnico Nacional, 3Postgrado en Ciencias
Genómicas, Universidad Autónoma de la Ciudad de México, 4Departamento de Matemáticas, 5Departamento de
Investigaciones Científicas y Tecnológicas, Universidad de Sonora, 6Department of Chemical and Enviromental
Engineering University of Arizona, 7Unidad de Investigación Médica e Inmunología, Instituto Mexicano del
Seguro Social, 8Centro de Investigaciones Regionales Hideyo Noguchi, Universidad Autónoma de Yucatán.
Abstract
DNA vaccines are one of the most promising health technologies for the prevention of infectious diseases. The direct
injection of DNA, such as naked plasmid DNA (pDNA) containing the sequence of a specific antigen into a living
host, causes that some of its cells produce small amount of the antigenic protein. This protein activates the specific
immune response of the host against the delivered antigen DNA. DNA vaccines have been identified among the
ten most important health biotechnologies for the improvement of the health world system in the present century.
In México, several groups have identified antigens with great potential for prevention and treatment of infectious
diseases, such as amebiasis, leishmaniasis, trypanosomiasis and rabies, among others, by pDNA vaccination.
Plasmid DNA immunization is potentially better, cheaper, and safer than the conventional protocols with mixture
or pure antigenic proteins. Nevertheless, one of the major limitations of DNA vaccines is the high amount of pure
and supercoiled pDNA required per dose. Once an antigen(s) is identified as good candidate in preclinical tests, the
amount of pDNA required for the next phase trial is technically and economically difficult to obtain by the standard
laboratory protocols. Therefore, a reproducible, safe, and economical process for the production and purification
of supercoiled pDNA should be developed. The aim of this study is to determine the conditions for a production of
pDNA vaccine at preparative scale using plasmid that already have been tested as potential DNA vaccines for the
leishmaniasis, amebiasis, Chagas disease and rabies. The goal of this research project is to improve the production,
isolation and purification of supercoiled pDNA with the quality required for clinical phase trial. Initial studies using
the pVAX-NH36, a pDNA vaccine candidate against leishmaniasis, have shown a 20-fold increase (from 2 to 40
mg/L) in flask and up to 60-fold (from 2-120 mg/L) in a 1 L batch bioreactor by only changing the Echerichia coli
growing medium. Experiments are in progress to optimize the medium, culture condition, as well as, the isolation
and purification of pDNA. The results of this work will help the public health system of Mexico City and the whole
country by contributing to the development of pDNA vaccines.
This work is supported by grant PIFUT08-108 (to J.O.L.) from Instituto de Ciencia y Tecnología del Distrito Federal.
*Contact
Phone: (55) 5747-3800 ext. 4381
e-mail: [email protected]
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Development of nanomaterials and biological vectors from viruses:
supramolecular assembly in vitro
Sánchez Rodríguez SP1, Echeverría OM2, Vázquez Nin GH2, Bustos Jaimes I1*
1Laboratorio de Fisicoquímica e Ingeniería de Proteínas, Facultad de Medicina, Universidad Nacional Autónoma
de México (UNAM), 2Laboratorio de Microscopía Electrónica, Facultad de Ciencias, UNAM.
Abstract
Viruses are supramolecular arrangements of nucleic acids enclosed into protein shells called capsids. Capsid-forming
proteins from different viruses can be expressed in bacteria or other simple organisms, producing thus empty capsids
also known as virus-like particles (VLPs). VLPs are valuable nanomaterials for chemical and biomedical applications,
as they are not infective. VLPs can be used in medicine as vaccines, imaging aids and molecular vectors for tissuespecific delivery of drugs or nucleic acids. The aim of our research is to set-up conditions for the in vitro assembly
and disassembly of VLPs of human parvovirus B19. Previous work has shown that the VP2 protein form this virus is
able to form VLPs in the absence of the other virus components, specifically other proteins and DNA. However this
is not enough for the use of this VLP in biomedicine. For the introduction of any kind of molecule into the formed
VLPs, it is mandatory to control the equilibrium between the assembled and disassembled states. Thus for the
introduction of molecules it is essential to have the proteins in its forming units, also called capsomers, while for the
field application it is required to shift the equilibrium to the VLP form of the protein.
Our work started by cloning the gene vp2, which codes for the VP2 protein, into a plasmid as expression vector.
The plasmid bearing the vp2 gene was introduced in the bacterium Escherichia coli. The vp2 gene expression was
induced and the recombinant protein purified under denaturizing conditions. The pure protein was refolded using
different conditions to form capsomers and VLPs. The formation of the different protein species was detected through
gel filtration chromatography. We are currently able to shift the equilibrium between the assembled and disassembled
forms of the VP2 protein. Our results clearly show that the formation of VLPs is highly biased by the physicochemical
conditions of the process. We have already obtained promising results from the encapsidation of model molecules.
Future experiments will show if these VLPs have potential as molecular vectors and nanoparticles.
*Contact
Phone: (55) 5623-2260
e-mail: [email protected]
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Bacteriophages as tools to characterize and control the opportunistic
clinical-pathogen Pseudomonas aeruginosa
Sepúlveda Robles O1, Martínez Peñafiel E1, Kameyama Kawabe L1,
Guarneros Peña G1*
1Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados-Instituto
Politécnico Nacional, México, D.F.
Abstract
The opportunistic pathogen Pseudomonas aeruginosa (Pa) is one of the major nosocomial problems in the world
due to its ubiquity, nutritional versatility and high resistance to antibiotics. In Mexico, 28.500 cases of nosocomial
infections are reported each year. Therefore, it is crucial to develop new methodologies for the rapid identification
of clinical strains of Pa to implement faster and more accurate treatment of infected patients. Also it is important
to propose alternatives to treat Pa infections and to monitor sources of pathogen dissemination in hospitals. To
accomplish these goals we are using viruses called bacteriophages or phages, which specifically infect and lyse Pa
strains. We have isolated and characterized 72 phages from different sources of natural reservoirs and waste-water
and assayed against 154 clinical isolates of Pa from different hospitals in Mexico City as hosts. All 72 phages infected
one or more Pa clinical strains to yield a specific pattern named host range. This method distinguished 67 unique
patterns of phage infection for strain identification. Most of the phages analyzed contained DNA as genetic material
and only 6 contained RNA. The DNA analysis using EcoRI and HindIII restriction enzymes showed that 53 out of 58
phages presented unique restriction profiles. In addition, we characterized the phage morphology for each case using
electron microscopy. Accordingly four morphological types were distinguished: 36 Siphoviruses, 12 Podoviruses, 7
Myoviruses and 6 Leviviruses. We have assembled a collection of phage and will attempt to typify Pa variants usually
observed in nosocomial outbreaks. The appropriate use of this tool will allow a rapid identification of Pa variants to
help making the therapeutic decisions concerning critically infected patients. Other epidemiological questions, such
as if the isolates of different malaises correlate with specific phage infection types will be addressed.
*Contact
Phone: (55) 5747-3800 ext. 5352
e-mail: [email protected], [email protected]
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Analysis of bacteriophages from Pseudomonas aeruginosa strains
prevalents in Mexico City hospitals
Uc-Mass A1, Guarneros Peña G1, Kameyama Kawabe L1*
1Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados-Instituto
Politécnico Nacional, México, D.F.
Abstract
The bacterium Pseudomona aeruginosa is a ubiquitous Gram-negative soil microorganism, it can growth in diverse
humid environments and is able to express a variety of virulent determinant that cause a wide range of infections in
plants, nematodes, insects and animals. Pseudomonas aeruginosa have medical importance because is able to persist
and causes intra-hospitalary infections; as a typical opportunist infects injured, burned and inmunodeficient humans
and can cause chronic respiratory infections in individuals with cystic fibrosis. In the genome of Pseudomonas
aeruginosa the prophage existency has been associated with virulent traits and with genome plasticity. Around the
29% of variability between the genomes of different strains is due to the presence of phage-like sequences. Due to
the relevancy of phages in the Pseudomonas virulence we analyzed tree clinical strains collection of Pseudomonas
aeruginosa to find prophages. We obtained 59 samples of phages from the tree clinical collections that were lysogens,
and purified their genomes. The profile of migration of the digested genomes was used to identify unique phages,
this criteria permit us to detect 23 new phages. Also, we examined the phage morphology by electron microscopy of
fifteen of them, and we found that thirteen belongs to the Siphoviridae family, and just two of them are Podoviridae.
From the 23 phages isolated just one of them was founded in the tree collections, and this phage was also the phage
isolated at higher frequency (ten times); another two phages were founded in two collections, and they were isolated
seven times. We are currently constructing lysogens to test the exclusion among these phages.
*Contact
Phone: (55) 5747-3800 ext. 5352
e-mail: [email protected], [email protected]
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Design optimization of the multiarticulated prosthetic robotic hand
Velázquez Sánchez AT1*, Hernández Gómez LH, Torres San Miguel CR, Merchán Cruz EA2, Mejía Domínguez
JA1, Escalante Rodríguez E3
1Escuela Superior de Ingeniería Mecánica y Eléctrica campus Zacatenco, Instituto Politécnico Nacional (IPN),
2Escuela Superior de Ingeniería Mecánica y Eléctrica campus Azcapotzalco, IPN, 3Hospital Regional 1ro de
Octubre, Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado, México, D.F.
Abstract
This work shows the optimization of mechanical design and construction of the “Multiarticulated prosthetic robotic
hand (ROBOHANDIPN)” by analyzing the hand structure, and the optimization of the mechanical components,
through to use of genetic algorithms and the artificial neural networks, to ensure that the elements don’t fail during
operation and identify the structural constraints of the system.
We need to optimize the design of underactuated mechanisms for this implementation in robotic prosthetic hand;
there is an analysis of the structural elements of the robotic hand in order to: Synthesize the design to ensure that
none of the elements fail during operation and to identify the functional limitations of the hand.
Research conducted previously on this subject and specifically the work presented by Dr. Velázquez; provide tools
on the behavior of the hand grip in specific situations, identifying the essential characteristics of this system.
1.- kinematics Optimization of the mechanical structure.
2.- Simulation and Construction.
3.-The integration of electronic and control components.
According to the background, can be seen that to date, have not developed national prosthesis that can provide
greater dexterity in handling objects as presented in the daily lives and maimed unlikely that a patient recovers. Is
necessary a more thorough analysis on the structural synthesis of mechanisms for multiarticulated robotic hands. The
investigations are focused on the hand in order to mimic the movements and to provide the prosthetic hand dexterity
and skill with which the human mind to manipulate objects. However, by increasing the number of fingers and joints
that have the mechanisms, it also increases the degree of control employed in the same, so it is necessary to conduct
a thorough structural optimization, in order to execute the main types grip by the hand.
Regarding the development of robotic hands nationwide, has made the design of grippers with appearance of the
hand, but not with anthropomorphic features, much less with the ability to perform different types of grip. These
clamps are used as a prosthetic hand, performed the opening and closing without ensuring a natural movement in
the fingers, and there is no independent movement of the same. There are few reported work related to the hand or
robotic anthropomorphic end effectors, and most of the work of the prosthetic hand designed primarily focusing on
obtaining an anthropomorphic appearance, although this is not multiarticulated, and does not have sufficient skill
for handling of various objects.
It is possible to synthesize a four-bar mechanism with kinematic coupling, able to emulate the movement of the index
finger to grab the cylinder and timely, based on the experimentally obtained kinematic variables and to extrapolate
this analysis to the other fingers to form a multiarticulated robotic hand.
*Contact
Phone: (55) 572-96000
e-mail: [email protected] , [email protected]
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