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Studies have identified that diseases in pregnancy affect fetal growth and development of the newborn. In Mexican population, the gene SLC16A11 has been identified as a factor that increases the risk of developing type 2 diabetes mellitus. To date, information is scarce about its expression in gestational diabetes mellitus (GDM); epigenetic modifications due to maternal hyperglycemic state could be identified early in fetal development.This study aimed to determine the SLC16A11 expression and methylation status in umbilical cord blood of newborns offspring of mothers with or without GDM.Cross-sectional, analytic study. Pregnant patients undergoing caesarean delivery with and without GDM in the Unidad Medica de Alta Especialidad Hospital de Gineco-obstetricia #4 Luis Castelazo Ayala, Instituto Mexicano del Seguro Social, were invited to participate. DNA was extracted from the mothers' blood cells, or umbilical cord blood cells of their newborns, and subjected to methylation status. Total RNA was used to evaluate the SLC16A11 expression by endpoint RT-PCR. Variables were analyzed with Student t. Values of p0.05 were considered statistically significant.A SLC16A11 downregulation was observed for newborns, while methylation status was found in only 1 of 68 mother-child pairs. Somatometry of newborns showed no differences between groups. Differences were found in total cholesterol, triglycerides, ALT, glucose, and HbA1c.For the first time, a differential expression for SLC16A11 was observed in offspring. Downregulation in this gene expression could characterize the offspring from GDM. No difference was found in somatometry of newborns of mothers with and without GDM.

We are currently witnessing a worldwide event caused by the pandemic outbreak derived from the new SARS-CoV-2 virus, which requires the generation of knowledge. Due to its novelty, many hypotheses and theories are discussed daily regarding the origin of this new virus. Several studies are focused on demonstrating how similar it is to other viruses.To highlight the differences of SARS-CoV-2 with other SARS viruses, from a comparative genomics analysis, and determine if these can be attributed to manipulation events.Complete genomes of two SARS viruses were downloaded, along with other six of human coronaviruses, and 16 of SARS-type coronaviruses. These were analyzed using the BLAST Ring Image Generator tool; afterwards, the evident differences were examined by MAFFT and BLAST programs.High identity was observed in fragments of the mammalian SARS-like genomes with the SARS-CoV-1 and SARS-CoV-2 genomes, identifying three main nucleotide differences, in the ORF1ab nsp3 region gene, in the receptor recognition S gene, and in the ORF8 gene, with which the SARS-type strains of mammals can be separated into the SARS-CoV-1 and SARS-CoV-2 type.The complete SARS-CoV-2 genome has high identity with mammalian SARS-type strains, which is why its most probable appearance could be the result of natural evolution.en este momento somos testigos de un evento de magnitud mundial provocado por el brote pandémico derivado del nuevo virus SARS-CoV-2, lo cual requiere la generación de conocimiento. Por lo novedoso que resulta, muchas hipótesis y teorías son discutidas a diario respecto al origen de este nuevo virus. Varios estudios están enfocados en demostrar la similitud que el SARS-CoV-2 tiene con otros virus.resaltar las diferencias del SARS-CoV-2 con otros virus SARS, a partir de un análisis de genómica comparativa, y determinar si se pueden atribuir a eventos de manipulación.se descargaron dos genomas completos de virus SARS, seis genomas completos de coronavirus humanos y 16 de coronavirus tipo SARS; fueron analizados en un estudio de genómica comparativa mediante la herramienta BLAST Ring Image Generator, y a continuación se examinaron las diferencias evidentes mediante el uso de los programas MAFFT y BLAST.se observó una alta identidad en fragmentos de los genomas tipo SARS de mamíferos con los genomas SARS-CoV-1 y SARS-CoV-2, y se identificaron tres diferencias nucleotídicas principales: en el gen ORF1ab región nsp3, en el gen S de reconocimiento al receptor y en el gen ORF8, con el cual se pueden separar las cepas tipo SARS de mamíferos en tipo SARS-CoV-1 y SARS-CoV-2.el genoma completo de SARS-CoV-2 posee una alta identidad con cepas tipo SARS de mamíferos, por lo que su aparición más probable podría ser el resultado de la evolución natural.

espanolEn el afan por explicar la fisiopatogenia de COVID-19 se estan realizando estudios en torno a la correspondencia entre la expresion de receptores celulares de SARS-CoV-2 y las secuencias virales. La expresion de los receptores ACE2, CD147 y TMPRSS2 podria indicar blancos de infeccion poco explorados. Para el analisis genomico de los receptores de SARS-CoV-2 se opto por utilizar la informacion del BioGPS, un portal que centraliza los recursos de anotacion genetica, en combinacion con la de The Human Protein Atlas, el portal mas grande de datos del transcriptoma y proteoma humanos. Tambien se revisaron los articulos mas recientemente respecto al tema. En numerosos sitios anatomicos se observo la expresion de ARN y proteinas de los receptores del virus, que coinciden parcialmente con la informacion reportada en la literatura. Resalto la alta expresion en las celulas de los testiculos, por lo que seria importante descartar si este sitio anatomico es un reservorio de SARS-CoV-2; de no ser asi, determinar el dano en las celulas germinales, tal como sucede en infecciones por otros virus ARN. EnglishIn the efforts to explain COVID-19 pathophysiology, studies are being carried out on the correspondence between the expression of SARS-CoV-2 cell receptors and viral sequences. ACE2, CD147 and TMPRSS2 receptors expression could indicate poorly explored potential infection targets. For the genomic analysis of SARS-CoV-2 receptors, using BioGPS information was decided, which is a portal that centralizes genetic annotation resources, in combination with that of The Human Protein Atlas, the largest portal of human transcriptome and proteome data. We also reviewed the most recent articles on the subject. RNA and viral receptor proteins expression was observed in numerous anatomical sites, which partially coincides with the information reported in the literature. High expression in testicular cells markedly stood out, and it would be therefore important ruling out whether this anatomical site is a SARS-CoV-2 reservoir; otherwise, germ cell damage, as it is observed in infections with other RNA viruses, should be determined.

In the efforts to explain COVID-19 pathophysiology, studies are being carried out on the correspondence between the expression of SARS-CoV-2 cell receptors and viral sequences. ACE2, CD147 and TMPRSS2 receptors expression could indicate poorly explored potential infection targets. For the genomic analysis of SARS-CoV-2 receptors, using BioGPS information was decided, which is a portal that centralizes genetic annotation resources, in combination with that of The Human Protein Atlas, the largest portal of human transcriptome and proteome data. We also reviewed the most recent articles on the subject. RNA and viral receptor proteins expression was observed in numerous anatomical sites, which partially coincides with the information reported in the literature. High expression in testicular cells markedly stood out, and it would be therefore important ruling out whether this anatomical site is a SARS-CoV-2 reservoir; otherwise, germ cell damage, as it is observed in infections with other RNA viruses, should be determined.

"Introducción: en este momento somos testigos de un evento de magnitud mundial provocado por el brote pandémico derivado del nuevo virus SARS-CoV-2, lo cual requiere la generación de conocimiento. Por lo novedoso que resulta, muchas hipótesis y teorías son discutidas a diario respecto al origen de este nuevo virus. Varios estudios están enfocados en demostrar la similitud que el SARS-CoV-2 tiene con otros virus. Objetivo: resaltar las diferencias del SARS-CoV-2 con otros virus SARS, a partir de un análisis de genómica comparativa, y determinar si se pueden atribuir a eventos de manipulación. Material y métodos: se descargaron dos genomas completos de virus SARS, seis genomas completos de coronavirus humanos y 16 de coronavirus tipo SARS; fueron analizados en un estudio de genómica comparativa mediante la herramienta BLAST Ring Image Generator, y a continuación se examinaron las diferencias evidentes mediante el uso de los programas MAFFT y BLAST. Resultados: se observó una alta identidad en fragmentos de los genomas tipo SARS de mamíferos con los genomas SARS-CoV-1 y SARS-CoV-2, y se identificaron tres diferencias nucleotídicas principales: en el gen ORF1ab región nsp3, en el gen S de reconocimiento al receptor y en el gen ORF8, con el cual se pueden separar las cepas tipo SARS de mamíferos en tipo SARS-CoV-1 y SARS-CoV-2. Conclusión: el genoma completo de SARS-CoV-2 posee una alta identidad con cepas tipo SARS de mamíferos, por lo que su aparición más probable podría ser el resultado de la evolución natural."

BACKGROUND: Studies have identified the intrauterine environment as a regulating factor of the methylation profiles of DNA of newborns. In Mexicans a SLC16A11 risk haplotypes is associated with type 2 diabetes mellitus. It is in our interest to evaluate SLC16A11 expression from the earliest stages of human development in the cells of umbilical cord blood of newborn offspring of mothers with and without gestational diabetes mellitus (GDM), and thus identify indirectly epigenetic modifications due to maternal hyperglycemic status. To date is scarce this kind of studies. OBJECTIVE: To determine the SLC16A11 gene expression, in the cells of umbilical cord blood of newborn offspring of mothers with or without GDM. MATERIAL AND METHODS: Pregnant patients submitting to cesarean, with or without GDM, were invited to participate in at Gyneco-obstetric hospital of IMSS, Mexico City. RNA of peripheral blood cells (PBC) from mothers with or without GDM, and from the umbilical cord blood cells of their newborn offspring, were extracted. Briefly, to evaluate differences in SLC16A11 gene expression, cDNA was generated by using retro-transcriptase reaction and then subjected to end point PCR with specific oligonucleotides. Variables were analyzed with t-Student test using the SPSS program. Values of p < 0.05 were considered statistically significant. RESULTS: Mean age of mothers was 30 ± 4.25 years. GDM mothers were heavier (32.90 ± 3.7 vs 30.12 ± 3.4 kg/m2, p=0.032), but there were no differences in glycemic control (HbA1c)(5.4 ± 0.6 vs 5.2 ± 0.6, p=0.333). Somatometric measurements of offspring showed no significant differences (weight: 3013.33 ± 675.54 vs 3135.71g ± 740.59, p=0.626 and length: 48.50 ± 3.03 vs 49.17cm ± 1.56, p=0.368). Analysis SLC16A11 gene expression did not showed differences in mothers, (p=0.059), and in offspring (p=0.315). Because there is a SLC16A11 gene expression, this could suggest that the SLC16A11 gene promoter could not be methylated. CONCLUSIONS: No modifications in SLC16A11 gene expression were present in GDM, which suggests that GDM does not modify the molecular mechanism in SLC16A11 RNA expression.

Worldwide, breast cancer (BrCa) is currently the leading cause of deaths associated to malignant lesions in adult women. Given that some studies have mentioned that peritumoral adipocytes may contribute to breast carcinogenesis, present work sought to quantitative evaluate the morphometry of these cells in a group of adult women. Three thousand six hundred sixty four breast adipocytes, that came from biopsies of a group of adult females with different types of breast carcinomas (ductal, lobular, and mixed) and one with normal tissues, were evaluated through an image analysis (IA) process regarding six morphometric descriptors: area (A), perimeter (P), Feret diameter (FD ), aspect ratio (AR), roundness factor (RF), and fractal dimension of cellular contour (FDC ). Data showed that the adipocytes of the normal tissues group were bigger (A: 3398 ± 2331 µm2 , P: 239 ± 83 µm, and FD : 79.9 ± 24.5 µm) than those from BrCa samples (A: 2860 ± 1933 µm2 , P: 214 ± 66 µm, and FD : 73.2 ± 22.5 µm), and presented a more irregular contour (FDC of 1.370 ± 0.037 for normal group and of 1.335 ± 0.049 for the oncologic one). Moreover, it could be accounted that adipocytes of mixed carcinomas were largest (FD : 75.1 ± 22.4 µm) than those of lobular lesions (FD : 61.6 ± 22.6 µm), while the adipocytes of ductal carcinomas were the most oval (AR: 1.421 ± 0.524) and roughest (FDC : 1.324 ± 0.050) cells. IA results suggest that BrCa lesions can be categorized through a quantitative morphometric evaluation of peritumoral adipocytes. These findings could let the development of an analytical tool to help the Pathologist to enhance the accuracy of the oncologic diagnose.

Hodgkin lymphoma (HL) is a rare neoplasm of the lymphatic system, in which inflammation and allelic variants in cytokines have been proposed as etiological factors. Epstein-Barr virus infection is often associated as a risk factor in HL and since cytokines are involved in the humoral response to viral infection. Our aim was to study the association between single nucleotide polymorphisms (SNPs) located in the tumor necrosis factor (TNF) gene (- 376GA, - 238GA and 581GA) in a sample of Mexican patients (56 cases) and their susceptibility to develop HL, comparing these SNPs among healthy individuals (127 controls). Frequencies for TNF - 238GA and TNF 581GA showed no significant differences between cases and controls. However, the proportion of cases with the GA genotype of - 376 SNP showed a significant difference as compared to controls, odds ratio = 4.41 (95% confidence interval: 1.21-16.6), p = 0.02. We found that in this group of patients from Mexico the SNP - 376GA in TNF shows an association with higher risk for HL.

Infection with human papillomavirus (HPV), mainly HPV type 16, is the major etiologic factor associated with cervical cancer (CC), but HPV infection alone is not sufficient for progression of precursor lesions. Host genetic susceptibility may lead to abnormal immune response resulting from virus persistence. Several studies have suggested a possible association with specific human leukocyte antigen (HLA) class I and II alleles and CC, but results are not consistent. The association of genetic HLA class I (A and B) and HLA class II (DR*B1 and DQ*B1) haplotypes with HPV16-positive CC (n = 104) and base population controls (n = 104) was evaluated in this Mexican population study. Sequence-specific primer HLA genes were determined by polymerase chain reaction (PCR)-based methods in peripheral blood cell counts (PCR sequence-specific oligonucleotides). The cervical swabs of 208 women were tested for HPV16 by Hybrid Capture II. Allele and haplotype HLA frequencies, Hardy-Weinberg tests, and a haplotype homogeneity test were estimated using the Arlequin software v. 3.01. Odds ratio (OR) was calculated to compare cases and control women. Consistent associations across other studies in women with CC and infected by HPV16 were observed for HLA-DRB1*15 (OR, 3.9; 95% CI, 1.6-10.2) and the haplotype DRB1*15 DQB1*0602 (OR, 4.1; 95% CI, 1.4-12.7) compared with control women. The HLA-A2-B44-DR4-DQ*0302, HLA-A24-B35-DR16-DQ*0301, and HLA-A2-B40-DR4-DQ*0302 haplotypes showed a positive association with CC (OR, >1), whereas HLA-A2-B39-DR4-DQ*0302, HLA-A24-B35-DR4-DQ*0302, and HLA-A68-B40-DR4-DQ*0302 showed a negative association (OR, Abbreviations:Cervical Cancer-CC, confidence interval-CI, human leukocyte antigens-HLA, human papillomavirus-HPV, odds ratio-OR, polymerase chain reaction-PCR, relative risk-RR, relative light units-RLU, ribonucleic acid-RNA, sequence-sensitive oligonucleotide-SSO

Introducción: las lesiones intraepiteliales escamosas (LIE) son un estado de transición hacia el cáncer cervicouterino (CaCu) y un déficit de micronutrientes puede acelerar este proceso. Por ello, determinar la existencia de este déficit y conocer qué factores se asocian permitiría una posible prevención en esta población de riesgo. Objetivo: determinar la presencia de alguna deficiencia de micronutrientes involucrados en el proceso anticancerígeno y asociar este déficit con hábitos y factores demográficos en pacientes con LIE de Ciudad Juárez, Chihuahua, México.Métodos:en un estudio transversal analítico fueron seleccionadas 102 pacientes con LIE. Se realizó una encuesta dietaría (recordatorio de 24 horas) para estimar la ingesta de micronutrientes. La deficiencia fue determinada con un consumo < 75% de la ingesta diaria recomendada o sugerida (IDR o IDS) en México. Algunos hábitos y factores demográficos fueron obtenidos mediante la entrevista con la paciente. Se realizó un modelo de regresión logística para asociar la presencia de deficiencia con factores que afectan a la ingesta o incrementan el requerimiento de micronutrientes. Resultados:el retinol, ácido fólico, zinc, vitaminas C y E, considerados como micronutrientes en el proceso anticancerígeno del CaCu, se encontraron por debajo del 75% de la IDR. Aquellas mujeres con sobrepeso, obesidad y amas de casa se asociaron significativamente con la deficiencia de micronutrientes. Conclusión: el sobrepeso, la obesidad y la ocupación han sido asociados para presentar deficiencias de micronutrientes en este estudio. Estas variables convergen en una posible inseguridad alimentaria, la cual podría asociarse al incremento de incidencia de CaCu en México.

We developed a procedure to detect the 7 point mutations at Cys634 of the proto-oncogene RET, which is responsible for medullary thyroid carcinoma (MTC). Genomic DNA was prepared from blood samples obtained from normal and MTC-affected individuals belonging to a family with a history of the disease. The RET genotype for each individual was first established by performing restriction and sequencing analyses. Single-stranded target DNA was prepared by asymmetric polymerase chain reaction (PCR) amplification of a 93-bp fragment containing Cys634. The target was annealed with pairs of prelabeled stacking oligonucleotides designed to create appropriate 7-nucleotide gaps, which served as the sites of subsequent hybridization with glass-immobilized 7-mer probes. The target-stacking oligonucleotide duplexes were hybridized with DNA chips containing a set of eight 7-mer probes designed to detect the wild-type sequence and the seven point mutations described. We tested two sets of immobilized probes containing internal or 5'-terminal codon-634 single-base variations. Both groups of probes were able to discriminatively identify the mutations. The hybridization patterns indicated that the disease in this family was due to the C634Y mutation, in accord with the original sequence analysis. The hybridization-based mutation assignment was additionally supported by determination of the control homozygous and heterozygous hybridization patterns produced with synthetic targets having the normal or codon 634 mutant sequences. The effects of mismatch type and nearest-neighbor sequences on the occurrence of false-positive (mismatched) hybridizations are discussed.

Background. The invasive and metastatic potential of malignant cells results from complex interactions of numerous factors not yet fully understood. Genomic alterations such as ras overexpression and nm23-H1 inhibition have been found to be frequently associated with increased invasiveness in various cancers. On the other hand, secretion of different proteinases are necessary for malignant cells to traverse a network of matrix macromolecules, but the relationship between the genomic alterations and the proteolytic phenotype is still unclear. Our aim was to investigate whether the appearance of the proteolytic phenotype had any correlation with the expression of H-ras and nm23-H1 genes in carcinoma of the uterine cervix. Methods. Twenty-five samples from patients with carcinoma of the uterine cervix at different clinical stages were studied. Cathepsin B1, plasminogen activator, and collagenase activity were assessed in tissue cytosols using specific synthetic oligopeptides as substrates. The expression of H-ras and nm23-H1 was investigated by means of immunohistochemistry and in situ hybridization. Results. Our results showed that cathepsin B1 was the most consistently elevated proteinase, demonstrating a linear correlation with clinical staging. H-ras expression was found elevated in 40% of the cases. Nm23-H1 protein immunoreactivity was positive in 40% of the cases. No correlation was found among H-ras, cathepsin B1 activity, and survival rate. Among cases with high cysteine proteinase activity, a different clinical behavior depending on the expression of Nm23-H1 was observed. The cases with Nm23-H1 protein had a markedly better survival rate than those lacking this protein. In contrast, the absence of Nm23-H1 in association with high cathepsin B1 activity was a clear indicator of a poor prognosis. Conclusions. These findings suggest a complex interaction between the proteolytic phenotype and the expression of H-ras and nm23-H1 genes in carcinoma of the cervix that influences the clinical behavior of the tumor. © 2000 IMSS. Published by Elsevier Science Inc.

Cervical cancer and cervical intraepithelial neoplasia (CIN) require a careful selection of conventional and complementary therapies. The inclusion of dietary supplements within the aforementioned treatments set the tone within treatments. However, there are no reports of what kind of supplements could be used to provide a better response to the disease in these patients.In this review, we analyze clinical trials of the past 20 years that evaluated dietary supplements on this population, with the objective to raise awareness on which of them are viable to be administered.We performed a search for clinical trials that used a dietary supplement in women with cervical cancer and some degree of NIC analyzing them how to use the supplement, the conventional treatment, deficiency of the nutrient or compound to be evaluated, the characteristics of the population, the clinical trial and the supplement, as well as dosage and their effects.Twenty were in total the number of studies reviewed. The articles were classified according to the nature of the supplement: Retinoids, vitamin E, probiotics, indoles, multivitamin, folic acid and selenium.Some supplements were found to be effective in the treatment of cervical cancer and CIN. However, their effect depends on specific factors of the disease. The correct management of dietary supplements is an effective aid to help the patient with cancer and therefore, it is important to define what complementary therapies can be used for this population.Introducción: En el cáncer cervicouterino (CaCu) y neoplasias intraepiteliales cervicales (NIC) se requiere de una cuidadosa selección de terapias convencionales y complementarias, la inclusión de suplementos dietéticos dentro de ellas marcaría la pauta dentro de los tratamientos. Sin embargo, no se tiene algún reporte de qué tipo de suplementos pudieran ser utilizados para brindar una mejor respuesta ante la enfermedad por estas pacientes. Objetivo: En la presente revisión se analizan los ensayos clínicos de los últimos 20 años que evaluaron suplementos dietéticos en esta población, con el objetivo de dar a conocer cuáles de ellos son viables para ser administrados. Metodología: Se realizó una búsqueda de ensayos clínicos que utilizaron algún suplemento dietario en mujeres con CaCu y algún grado de NIC analizando en ellos el modo de utilización del suplemento; el tratamiento convencional; si existe una deficiencia del nutriente ó compuesto a evaluar; las características de la población, del ensayo clínico y del suplemento; así como la dosis y los efectos esperados. Resultados: Veinte fueron en total el número de estudios analizados. Los artículos fueron clasificados según la naturaleza del suplemento: Retinoides, vitamina E, probióticos, indoles, multivitamínico, ácido fólico y selenio. Conclusión: Para el tratamiento de CaCu y NIC resultaron efectivos algunos de los suplementos encontrados en esta revisión, sin embargo su efecto dependerá de diferentes factores propios de la enfermedad. Debido a esto, es necesario el manejo correcto de los suplementos dietéticos para poder ser utilizados eficientemente como tratamiento complementario en esta población.

Recently, technological advances have greatly increased, generating the development of nanotechnology, which is responsible for the design of structures and materials in the nanometer scale. This creates one of the most important cutting-edge sciences, integrating physics, chemistry, engineering and biology sciences. Specifically the integration with biology results in a new science called nanobiotechnology, specifically nanomedicine, which has the goal of mainly looking for more precise molecular diagnostic and prognostic processes, as well as the new design of drugs in the personalized medicine field. On the other hand, at molecular level in medical research, the nanoparticles are most commonly used as tools. Molecular diagnostics uses gold nanoparticles, paramagnetic nanoparticles and quantum dots, which can be used for the diagnosis and treatment of several diseases, including cancer. Quantum dots are the most promising tools for diagnosis and therapy in cancer research.

This study determined the methylation status of cellular retinoic acid-binding protein (CRABP) gene promoters and associated them with demographic characteristics, habits, and the presence of human papilloma virus (HPV) in patients with cervical cancer (CC), low and high squamous intraepithelial lesions, and no intraepithelial lesion. Women (n = 158) were selected from the Colposcopy Clinic of Sanitary Jurisdiction II in Ciudad Juarez, Chihuahua, Mexico. Demographic characteristics and habit information were collected. Cervical biopsy and endocervical scraping were used to determine methylation in promoter regions by methylation-specific polymerase chain reaction technique. We found hemi-methylation patterns in the promoter regions of CRABP1 and CRABP2; there was 28.5% hemi-methylation in CRABP1 and 7.0% in that of CRABP2. Methylation in CRABP1 was associated with age (≥35 years, P = 0.002), family history of cancer (P = 0.032), the presence of HPV-16 (P = 0.013), and no alcohol intake (P = 0.035). These epigenetic changes could be involved in the CC process, and CRABP1 has the potential to be a predictive molecular marker of retinoid therapy response.

Our objective was to determine the association between viral load of high risk human papilloma virus (HPV) using the Hybrid Capture II (HC II) system and cervical intraepithelial neoplasia (CIN) lesion stage.A total of 182 consecutive women with confirmed diagnoses of CIN 1-3 and 182 healthy women with negative Pap were included. All subjects underwent structured interviews focused on socioeconomic and reproductive factors. HC II testing was used to detect human papilloma virus (HPV) DNA. Viral load was measured by light measurements expressed as relative lights unit (RLU) ratio (specimens/control). Log(10)RLU ratios were categorized for analysis into four groups: negative (/=0); low viral load (0.01-1.0), middle viral load (1.01-2.0), and high viral load (2.0-3.6). Frequencies and association measurement odds ratio (OR) adjusted by unconditional multinomial regression (UMR) were used in analysis.A total of 75 of 80 (93.7%) patients with CIN 2-3, 82 of 101 (79.4%) with CIN 1, and 36 of 182 (19.8%) controls were positive for HPV DNA. The higher the viral load of HPV DNA infection observed, the higher the probability of being associated with stage of CIN (P0.001). Association between low viral load HPV and CIN 1 was 16.8 (7.2-39) compared with the highest association observed with high viral load and CIN 2-3 (OR(a) = 365.8, 94.7-1412). Both control and cases in the oldest women presented the highest viral load.We found high frequencies of HPV DNA in CIN 1 and in CIN 2-3 patients. A clear association between viral load of HPV DNA was determined by HC II assay and CIN stage.

Composite pheochromocytomas (CP) account for only 3% of all pheochromocytomas. We analyzed the clinical, immunohistochemical, ultrastructural, DNA content, and 634 ret mutation features in a 56-year-old Mexican woman with CP localized in the right adrenal gland and associated to a blood pressure of 140/90 mmHg. Clinical symptoms were absent after surgery. The tumor showed pheochromocytoma and neuroblastoma components. This dual phenotype was supported by light microscopy and corroborated by immunohistochemistry and ultrastructural findings. Flow cytometric analysis showed that both components were diploid. A genetic mutational analysis of the ret oncogene in exon 11 showed no 634 mutation. This case demonstrates the indolent behavior of neuroblastoma associated to a sporadic-type CP in an adult patient.

We have developed an oligonucleotide microarray (genosensor) utilizing a double tandem hybridization technique to search for 9 point mutations located in the most frequently altered codons of the TP53 gene. Isolated and multiplexed PCR products, 108 and 92 bp long, from exons 7 and 8, respectively, were obtained from 24 different samples. Single-stranded target DNA was then prepared from isolated or multiplexed PCR products, through cyclic DNA synthesis. Independent ssDNA's were annealed with the corresponding pairs of labeled stacking oligonucleotides to create partially duplex DNA having a 7-nt gap, which contains the sequence that will be interrogated by the capture probes forming double tandem hybridization. In the case of multiplexed ssPCR products, only two stacking oligonucleotides were added per target, therefore the gap for the PCR products having two consecutive codons to be interrogated in exon 7 was 12 nt long, so only single tandem hybridization was produced with these respective probes. 18 codon substitutions were found by DNA sequencing. In 13 of them a perfect correlation with the pattern of hybridization was seen (In 5 no signal was seen with the wt probe while a new signal was seen with the appropriate mutant probe, and in 8 more, as expected, no signal was seen with any probe due to the absence of the corresponding probe in the array). In 3 other cases a mutation was falsely suggested by the combination of the absence of the wild type signal along with a false signal in the other probe. In the other 2 cases the presence of the mutation was not detected due to the production of a false hybridization signal with the wild type probe. In both cases (false mutation or no mutation detected) relatively stable mismatched target/probe duplexes should be formed. These problems could be avoided by the addition of probes to improve the performance of the array. Our results demonstrate that a simple TP53 microarray employing short (7-mer) probes, used in combination with single or double tandem hybridization approach and a simple or multiplex target preparation method, can identify common TP53 missense mutations from a variety of DNA sources with good specificity.