Impact of APOE Alleles-by-Diet Interactions on Glycemic and Lipid Features- A Cross-Sectional Study of a Cohort of Type 2 Diabetes Patients from Western Mexico: Implications for Personalized Medicine.

PURPOSE: To analyze clinically relevant interactions between the apolipoprotein E (APOE) ε2, ε3 and ε4 alleles and nutritional factors on glycemic control and lipid levels in a cohort of type 2 diabetes (T2D) patients from western Mexico. PATIENTS AND METHODS: In this cross-sectional study of the cohort of T2D patients, a total of 224 individuals were selected for interaction studies. Clinical and anthropometric data were obtained from pre-designed medical records. Dietary intake was assessed by validated three-day food consumption records. Biochemical measurements were determined by automated methods. APOE genotyping was performed by a real-time allelic discrimination assay. Gene-diet interactions were tested by corrected multiple linear regression analyses, which were adjusted by potential confounding factors such as age, sex, energy intake, BMI and anti-hyperglycemic therapy (Metformin, Glibenclamide or Insulin), and years with T2D. RESULTS: Seventy-six percent of patients with T2D were on Metformin therapy. The frequencies of the APOE alleles were ε2 (5.8%), ε3 (74.1%) and ε4 (20.1%). After statistical settings, significant APOE alleles-by-diet interactions in relation to the metabolic profile were found. Interestingly, higher blood levels of total cholesterol (p int. = 0.016), non-HDL-c (p int. = 0.024), and LDL-c (p int. = 0.030) were found only in carriers of the APOE ε2 allele with a low consumption of MUFA. In contrast, carriers of the APOE ε4 allele with a high ω-6:ω-3 PUFA ratio in the diet had higher %HbA1c blood concentrations (p int. = 0.035). CONCLUSION: This study suggests a differential metabolic impact of APOE alleles on lipid/glycemic phenotypes depending on the dietary intake, with important potential implications in the personalized medicine and nutritional management of patients with type 2 diabetes mellitus.

Interactions between DRD2/ANKK1 TaqIA Polymorphism and Dietary Factors Influence Plasma Triglyceride Concentrations in Diabetic Patients from Western Mexico: A Cross-sectional Study.

This study aimed to screen relevant interactions between DRD2/ANKK1 TaqIA polymorphism and dietary intakes with reference to phenotypical features in patients with T2D from western Mexico. In this cross-sectional study, a total of 175 T2D patients were enrolled. Dietary intake was evaluated using 3-day food records and appropriate software. Glycemic and blood lipid profiles were measured by standardized methods. Genotyping of the DRD2/ANKK1 TaqIA polymorphism was performed by the RFLP method. Gene-diet interactions regarding anthropometric and metabolic phenotypes were screened by adjusted multiple linear regression analyses. Genotype frequencies of the DRD2/ANKK1 TaqIA polymorphism were A1A1 (16.0%), A1A2 (52.6%), and A2A2 (31.4%). Statistically significant interactions between the DRD2/ANKK1 TaqIA genotypes and dietary factors in relation to blood triglyceride (TG) levels were found. Carriers of the A1 allele (A1A1 homozygotes plus A1A2 heterozygotes) were protected from TG increases by maltose intake (P int. = 0.023). Instead, A2A2 homozygotes were susceptible to TG rises through consumptions of total fat (P int. = 0.041), monounsaturated fatty acids (P int. = 0.001), and dietary cholesterol (P int. = 0.019). This study suggests that the interactions between DRD2/ANKK1 TaqIA polymorphism and dietary factors (sugar and fats) influence TG levels in diabetic patients.

Genotoxic Effect in Autoimmune Diseases Evaluated by the Micronucleus Test Assay: Our Experience and Literature Review.

Autoimmune diseases (AD) are classified into organ-specific, systemic, and mixed; all forms of AD share a high risk for cancer development. In AD a destructive immune response induced by autoreactive lymphocytes is started and continues with the production of autoantibodies against different targets; furthermore apoptosis failure and loss of balance in oxidative stress as a consequence of local or systemic inflammation are common features seen in AD as well. Micronucleus (MN) assay can be performed in order to evaluate loss of genetic material in a clear, accurate, fast, simple, and minimally invasive test. The MN formation in the cytoplasm of cells that have undergone proliferation is a consequence of DNA fragmentation during mitosis and the appearance of small additional nuclei during interphase. The MN test, widely accepted for in vitro and in vivo genotoxicity research, provides a sensitive marker of genomic damage associated to diverse conditions. In here, we present a review of our work and other published papers concerning genotoxic effect in AD, identified by means of the MN assay, with the aim of proposing this tool as a possible early biomarker for genotoxic damage, which is a consequence of disease progression. Additionally this biomarker could be used for follow-up, to asses genome damage associated to therapies.

Recombinant murine IL-12 promotes a protective Th1/cellular response in Mongolian gerbils infected with Sporothrix schenckii.

Sporotrichosis is a cutaneous fungal infection caused by Sporothrix schenckii. It is known to be mainly contained by Th1 responses. As IL-12 is crucial for Th1 response, we investigated if treatment with recombinant murine IL-12 (rmIL-12) promoted Th1 immunity and/or clinical improvement in an experimental sporotrichosis gerbil model. Gerbils were inoculated with S. schenckii in the footpad and treated with rmIL-12. Seven days post infection there was a significant increase in macrophage phagocytosis and oxidative burst, and in delayed-type hypersensitivity (DTH) reaction in rmIL-12 treated gerbils, as well as a ∼10-fold increase of serum IFN-gamma and a decrease of IL-4 and IL-10. Moreover, rmIL-12 substantially decreased (∼70%) S. schenckii burden in liver and spleen and improved the clinical outcome preventing footpad ulcer and tail nodules observed in untreated gerbils. Our study demonstrates that rmIL-12 promotes Th1 immune response against S. schenckii favouring its clearance and preventing clinical symptoms.

Micronuclei and other nuclear anomalies in exfoliated buccal mucosa cells of Mexican women with breast cancer.

PURPOSE: Breast cancer (BC) is the most frequently diagnosed form of cancer and the leading cause of cancer-related deaths among females in the world. RESULTS of several studies showed that the genome of primary cancer patients (naive for any treatment) is unstable. The purpose of the present study was to evaluate the genomic instability in BC patients by means of buccal cells micronucleus (MN) cytome assay Methods: The frequencies of nuclear anomalies including MN, binucleates (BN), broken eggs (BE), condensed chromatin (CC), karyorrhexis (KR) and karyolysis (KL) were evaluated in exfoliated buccal mucosa cells of Mexican women with primary BC and healthy women. Buccal cells were collected from 21 BC patients (9 with stage I and 12 with stage II) and from 20 healthy females used as control group. RESULTS: The results of the evaluation of cells showed that the frequencies of MN, BN, BE, KR and KL were significantly increased in the pooled group of BC patients compared with the control group. However, no one parameter of buccal MN-cytome assay in patients with stage I BC was significant compared with controls and BC patients with stage II. CONCLUSION: Application of the buccal MN-cytome assay for the study of genomic instability in primary BC patients showed that both genotoxic and cytotoxic effects can be evaluated in such patients.

Potential uses, limitations, and basic procedures of micronuclei and nuclear abnormalities in buccal cells.

The use of biomarkers as tools to evaluate genotoxicity is increasing recently. Methods that have been used previously to evaluate genomic instability are frequently expensive, complicated, and invasive. The micronuclei (MN) and nuclear abnormalities (NA) technique in buccal cells offers a great opportunity to evaluate in a clear and precise way the appearance of genetic damage whether it is present as a consequence of occupational or environmental risk. This technique is reliable, fast, relatively simple, cheap, and minimally invasive and causes no pain. So, it is well accepted by patients; it can also be used to assess the genotoxic effect derived from drug use or as a result of having a chronic disease. Furthermore the beneficial effects derived from changes in life style or taking additional supplements can also be evaluated. In the present paper, we aim to focus on the explanation of MN test and its usefulness as a biomarker; we further give details about procedures to perform and interpret the results of the test and review some factors that could have an influence on the results of the technique.

Interleucina-12 exógena evita disminución de actividad de mieloperoxidasa de macrófagos en gérbiles con esporotricosis / Exogenous interleukin-12 prevents the decrease in myeloperoxidase activity of macrophages in gerbils with sporotrichosis

Introducción: la interleucina-12 es una citosina inmunorreguladora con múltiples funciones biológicas, incluyendo la activación de macrófagos. La mieloperoxidasa es una enzima que desempeña un papel importante en la función antimicrobiana de fagocitos activados. En presencia de peróxido de hidrógeno, esta cataliza la oxidación de cloruro para generar ácido hipocloroso, potente agente microbicida. Objetivo: determinar el efecto estimulador de interleucina-12 recombinante murina sobre la actividad de mieloperoxidasa en macrófagos peritoneales en gérbiles infectados experimentalmente con levaduras de Sporothrix schenckii. Métodos: 500 ng de interleucina-12 recombinante murina fueron suministrados diariamente por vía intraperitoneal durante 5 días consecutivos a gérbiles machos, los cuales al sexto día fueron inoculados por vía subcutánea en el cojinete plantar posterior izquierdo con 6x10(6) levaduras de S. schenckii. siete días después de la infección los macrófagos fueron obtenidos de la cavidad peritoneal y la actividad de su mieloperoxidasa fue determinada mediante el método de Kaplow, expresándose como porcentaje de actividad de macrófagos peritoneales. Los resultados son expresados como el promedio del por ciento de actividad de mieloperoxidasa ± desviación estándar de 3 experimentos independientes. Las diferencias estadísticas entre grupos fueron evaluadas por medio de t de Student y un valor de p < 0,05 fue considerado significativo. Resultados: la administración de interleucina-12 recombinante murina a gérbiles previa infección con S. schenckii aumentó significativamente la actividad de mieloperoxidasa de macrófagos peritoneales (p = 0,0001) comparada con los controles sanos. En contraste, macrófagos de gérbiles infectados no tratados mostraron actividad de mieloperoxidasa significativamente disminuida comparada con los controles sanos (p= 0,001), lo cual sugiere activación deteriorada de macrófagos. Conclusiones: la administración in vivo de interleucina-12 recombinante murina antes de la infección con S. schenckii, induce activación de macrófagos evidenciada por un aumento en la actividad de mieloperoxidasa, lo que contribuiría a la defensa del organismo contra este agente infeccioso vía sistema oxidativo dependiente de mieloperoxidasa.

Introduction: interleukin-12 is an immunoregulatory cytokine with multiple biologic functions, including macrophage activation. Myeloperoxidase is an enzyme that plays an important role in the antimicrobial function of activated phagocytes. In the presence of hydrogen peroxide, myeloperoxidase catalyzes chloride oxidation to produce hypochlorous acid, a powerful microbicidal agent. Objective: determine the stimulating effect of murine recombinant interleukin-12 on myeloperoxidase activity in peritoneal macrophages of gerbils experimentally infected with Sporothrix schenckii yeasts. Methods: 500 ng murine recombinant interleukin-12 were administered intraperitoneally on a daily basis on 5 consecutive days to male gerbils. On the sixth day the gerbils were inoculated subcutaneously on the left posterior plantar pad with 6x10(6) S. schenckii yeasts. Seven days after infection, macrophages were obtained from the peritoneal cavity. Myeloperoxidase activity was determined by Kaplow's method and expressed as percentage of peritoneal macrophage activity. Results are expressed as the average percentage of myeloperoxidase activity ± standard deviation from 3 independent experiments. Statistical differences between groups were evaluated by Student's t test. A value of p < 0.05 was considered significant. Results: administration of murine recombinant interleukin-12 to gerbils following infection with S. schenckii significantly increased the myeloperoxidase activity of peritoneal macrophages (p = 0.0001) in comparison with healthy controls. Macrophages of untreated infected gerbils, however, showed significantly reduced myeloperoxidase activity in comparison with healthy controls (p= 0.001), suggesting poor macrophage activation. Conclusions: in vivo administration of murine recombinant interleukin-12 before infection with S. schenckii induces macrophage activation evidenced by an increase in myeloperoxidase activity, enhancing the organism's defense against that infectious agent via the myeloperoxidase-dependent oxidative system.

Lack of effects of the TNF-alpha and IL-10 gene polymorphisms in Mexican patients with lepromatous leprosy.

Several human genetic variants have been associated with susceptibility or resistance to leprosy. The aim of this study was to assess whether gene polymorphisms of -308 G/A TNF-alpha and -819 T/C IL-10 are associated with lepromatous leprosy in Mexican mestizos patients from northwest Mexico. We genotyped these polymorphisms by means of polymerase chain reaction (PCR) and restriction fragment length polymorphisms (RFLPs) in 68 patients with lepromatous leprosy and 144 healthy Mexican Mestizos controls. We found that the -308G TNF-alpha allele was predominant in both cases (94.3%) and controls (92.3%) without statistical significance and the frequencies of -819C IL-10 allele were also similar for the cases (56.0%) and controls (59.0%). These negative findings suggest that other genes or polymorphisms may be important in the susceptibility to leprosy infection in the Mexican mestizos.

Síndrome de lipodistrofia en pacientes con VIH/sida que reciben terapia antirretroviral de gran actividad en Tepic, México / Lipodystrophy syndrome in HIV/AIDS patients receiving highly active antiretroviral therapy in Tepic, Mexico

Objetivo: determinar la relación entre la manifestación del síndrome de lipodistrofia y la terapia antirretroviral de gran actividad en pacientes con VIH/sida. Métodos: se realizó un estudio descriptivo, transversal y correlacional en pacientes con VIH/sida que reciben terapia antirretroviral de gran actividad, atendidos entre marzo y diciembre de 2007 en el Centro Ambulatorio de Prevención y Atención del Sida e Infecciones de Transmisión Sexual (CAPASITS) de Tepic, Nayarit, México. La definición y diagnóstico del síndrome se realizó mediante el método de The National Centre in HIV Epidemiology and Clinical Research (NCHECR) de Australia. Se utilizó la prueba de chi cuadrado para evaluar la dependencia entre el síndrome y la terapia. Se evaluaron 175 pacientes (128 hombres y 47 mujeres), de 19 a 72 años de edad. Resultados: se diagnosticaron 141 pacientes (80,6) con síndrome de lipodistrofia (IC95% 74,7-86.4 %); el 82,6 % correspondió a hombres y el 74,5 % a mujeres. Según la severidad, el porcentaje fue de 17 % de grado 1, 3 % de grado 2, 10 % de grado 3 y 51 % de grado IV. Las pruebas de chi cuadrado para evaluar dependencia entre el síndrome y la terapia resultaron no significativas. Conclusiones: el síndrome de lipodistrofia severo resulta un serio problema para la apariencia de pacientes de VIH/sida que reciben o no la terapia antirretroviral y que agrega un riesgo cardiovascular importante que debe ser considerado para intentar su prevención o tratamiento. Aunque los resultados pueden presentar sesgos y limitaciones, aportan una aproximación importante para sustentar y planificar intervenciones sanitarias que disminuirían el impacto del síndrome en la salud de los pacientes con VIH/sida.

Objective: to establish the relationship between lipodystrophy syndrome and highly active anti-retroviral therapy in HIV/AIDS patients. Methods: a cross-sectional, descriptive and co-relational study was conducted in HIV/AIDS patients, who had received highly active antiretroviral therapy in Centro Ambulatorio de Prevención y Atención del Sida e Infecciones de Transmisión Sexual (CAPASITS) de Tepic in Nayarit, Méjico from March to December, 2007. The definition and diagnosis of this syndrome was based on the method developed by the National Center in HIV Epidemiology and Clinical Research (NCHECR) method in Australia. The Chi square test evaluated the dependence between syndrome and therapy. One hundred seventy five patients (128 men and 47 women) aged 19-72 years were evaluated. Results: one hundred forty one patients (80.6 %) were diagnosed with lipodystrophy syndrome (CI95%74,7-86,4 %); 82.6 % corresponded to men and 74.5 % to women. According to severity, 17 % classified as grade 1; 1.3 % in grade 2; 10 % in grade 3 and 51 % as grade IV. The Chi square tests for the evaluation of dependence between syndrome and therapy were not significant. Conclusions: severe lipodystrophy syndrome is a serious health problem for HIV/AIDS patients receiving antiretroviral therapy or not. It adds a significant cardiovascular risk that must be considered for prevention or treatment. Although the results may be biased or restricted, they represent an important approach to planning and performing health interventions that will reduce the impact of the syndrome on the health of HIV/AIDS patients.

Topical pimecrolimus lacks genotoxicity and cytotoxicity by means of micronucleus erythrocyte rodent assay.

Topical pimecrolimus is an alternative treatment of atopic dermatitis. However, rare cases of malignancy have been reported with their use. This study was performed to investigate the possible geno- or cytotoxic effect in mouse bone marrow caused by systemic absorption of pimecrolimus 1% cream. In order to determine this, induction of micronucleated erythrocytes (MNE) in mouse peripheral blood was determined after the cutaneous application of three different doses, daily for 5 consecutive days. No differences were found in frequencies of polychromatic erythrocytes, MNE, and micronucleated polychromatic erythrocytes in the different groups of study. In conclusion, under described conditions, no geno- or cytotoxic effects were detected after the cutaneous application of pimecrolimus.

Induction of micronucleated erythrocytes in mouse peripheral blood after cutaneous application of 5-fluorouracil.

BACKGROUND: For topically applied drugs such as 5-fluorouracil (5-FU), dosage is not as precise as for other drug administration pathways. Consequently, quantity of drug delivered may differ among individuals and applications. 5-FU is used in treatment of different diseases and has been reported as a clastogenic compound by micronucleus assay. METHODS: To determine whether 5-FU cream (5% 5-FU) absorbed through skin can produce genotoxic or cytotoxic effect in mouse bone marrow, induction of micronucleated erythrocytes (MNE) in mouse peripheral blood was examined after cutaneous application of 5-FU daily for 5 days. RESULTS: 5-FU cream induced significant micronuclei at doses of 37.5 mg (total weight of cream)/2 cm(2) and 75.0 mg/2 cm(2), as well as cytotoxic effects at doses of 150.0 and 300.0 mg/2 cm(2). CONCLUSIONS: Cutaneous application of 5-FU increased number of MNE in mouse peripheral blood. These data emphasize the importance of using correct dose when applying drugs topically.

Chlamydia trachomatis infection in pregnant women with premature membrane rupture or premature delivery threat

Introducción. Chlamydia trachomatis se considera el agente causal de tracoma, salpingitis, endometritis y podría estar involucrada en la ruptura prematura de membrana (PMR) y amenaza de parto prematuro (PDT). El objetivo de este trabajo fue determinar la presencia de antígenos de C. trachomatis y anticuerpos contra C. trachomatis en mujeres embarazadas con PMR, PDT (ambos grupos de etiología desconocida) y mujeres con embarazo normal (NP).Material y métodos. Se obtuvieron 50 muestras endocervicales por cada grupo de mujeres embarazadas, para la determinación de antígenos de C. trachomatis, por el método de inmunofluorescencia directa. Asimismo fueron tomados 5 ml de sangre venosa, para identificar la presencia de anticuerpos contra C. trachomatis por inmunofluorescencia indirecta. Resultados. Seis por ciento (3/50) de las pacientes con PMR presentaron antígenos de C. trachomatis y anticuerpos IgG anti-C. trachomatis. Dos por ciento (1/50) con PDT tuvieron antígenos de C. trachomatis y anticuerpos IgM anti-C. trachomatis. Seis por ciento (3/50) de las pacientes con NP mostraron antígenos de C. trachomatis, pero no anticuerpos anti-C. trachomatis. Sin embargo, en 10 por ciento (5/50), 10 por ciento (5/50) y 16 por ciento (8/50) con PMR, PDT o NP, respectivamente; solamente se encontraron anticuerpos IgG anti-C. trachomatis. Conclusión. El hallazgo tanto de antígenos como anticuerpos anti-C. trachomatis en los tres grupos estudiados, resalta la importancia de la oportuna identificación de la infección, para la aplicación del tratamiento adecuado, para prevenir las secuelas de la infección, tanto en las mujeres embarazadas como en sus productos.