Proteomic Identification and Quantification of Basal Endogenous Proteins in the Ileal Digesta of Growing Pigs.

The accurate estimation of basal endogenous losses (BEL) of amino acids at the ileum is indispensable to improve nutrient utilization efficiency. This study used a quantitative proteomic approach to identify variations in BEL in the ileal digesta of growing pigs fed a nitrogen-free diet (NFD) or a casein diet (CAS). Eight barrow pigs (39.8 ± 6.3 kg initial body weight (BW)) were randomly assigned to a 2 × 2 crossover design. A total of 348 proteins were identified and quantified in both treatments, of which 101 showed a significant differential abundance between the treatments (p < 0.05). Functional and pathway enrichment analyses revealed that the endogenous proteins were associated with intestinal metabolic function. Furthermore, differentially abundant proteins (DAPs) in the digesta of pigs fed the NFD enriched terms and pathways that suggest intestinal inflammation, the activation of innate antimicrobial host defense, an increase in cellular autophagy and epithelial turnover, and reduced synthesis of pancreatic and intestinal secretions. These findings suggest that casein diets may provide a more accurate estimation of BEL because they promote normal gastrointestinal secretions. Overall, proteomic and bioinformatic analyses provided valuable insights into the composition of endogenous proteins in the ileal digesta and their relationship with the functions, processes, and pathways modified by diet composition.

Vaccination Strategies in a Potential Use of the Vaccine against Bovine Tuberculosis in Infected Herds.

Bovine tuberculosis (bTB) is a disease of cattle that represents a risk to public health and causes severe economic losses to the livestock industry. Recently, one of the strategies recommended for reducing the prevalence of the disease in animals is the use of the BCG vaccine, alone or in combination with proteins. It has been shown that the vaccine elicits a strong immune response, downsizes the number of animals with visible lesions, and reduces the rate of infection as well as the bacillary count. This paper, based on scientific evidence, makes suggestions about some practical vaccination alternatives that can be used in infected herds to reduce bTB prevalence, considering BCG strains, vaccine doses, routes of application, and age of the animals. Our conclusion is that vaccination is a promising alternative to be included in current control programs in underdeveloped countries to reduce the disease burden.

Changes in lactate dehydrogenase on admission throughout the COVID-19 pandemic and possible impacts on prognostic capability.

Introduction: The enzyme lactate dehydrogenase (LDH) is a good marker of general hyperinflammation correlated with mortality for COVID-19, and is therefore used in prognosis tools. In a current COVID-19 clinical randomized trial (CRT), the blood level of LDH was selected as an inclusion criterion. However, LDH decreased during the pandemic; hence, the impact of this decrease on the prognostic value of LDH for mortality was evaluated. Methods: Data on LDH levels in 843 patients were obtained and analyzed. Relative risk, standard error and receiver operating characteristic curves were calculated for two cutoff values. Results: Relative risk lost validity and the area under the curve narrowed by trimester during the pandemic. Conclusion: The progressive decrease in LDH impacted the capacity to predict mortality in COVID-19. More studies are needed to validate this finding and its implications.

A.L.L. Y.O.U. N.E.E.D. I.S. L.O.V.E. Manual on health self-management and patient-reported outcomes among low-income young adult Mexicans on chronic dialysis: Feasibility study.

PURPOSE: We evaluated disease knowledge/self-management skills among low-income Mexican young adults maintained on dialysis and to test the effectiveness of the A.L.L. Y.O.U. N.E.E.D. I.S. L.O.V.E (AYNIL) Manual - Spanish Version on patient-reported outcomes. This is a low literacy teaching tool designed with patients and educators' input. DESIGN AND METHODS: A quasi-experimental study was conducted in 17 chronic dialysis patients at Mexico City's Hospital General de México, Dr. Eduardo Liceaga. Ages 18-30-year-old completed disease knowledge/self-management and quality of life measures before the intervention and 6 weeks later. RESULTS: Significant increases were observed on disease knowledge/self-management scores in the STARx questionnaire from 47 (IQ: 40,51) to 50 (IQ: 48,54) p = 0.04. The UNC-TRxANSITION Index increased significantly from 4.8 (IQ: 3.9,5.7) to 7.7 (IQ: 7.5,8.2) p ≤0.001. Significant increases in scores were detected in the "Burden of kidney disease" (p = 0.008), "Effects of kidney disease" " (p = 0.03) and " Dialysis staff encouragement" (p = 0.027) based on the KDQoL survey. CONCLUSIONS: In this vulnerable population, the Spanish version of the A.L.L. Y.O.U. N.E.E.D. I.S. L.O.V.E. - AYNIL Manual improved CKD/ESRD disease knowledge/self-management skills and HRQoL. This study highlighted the need for low-literacy educational tools to improve patient-reported outcomes. PRACTICE IMPLICATIONS: Young adults with CKD/ESRD can benefit from patient-centered educational interventions to enhance their autonomy and the development of self-management behaviors that improve patient-reported outcomes and potential complications of the disease. Special attention is needed in low-income patients with low rates of adherence to treatments and poor self-management skills.

Prime Vaccination with Chitosan-Coated Phipps BCG and Boosting with CFP-PLGA against Tuberculosis in a Goat Model.

Attempts to improve the immune response and efficacy of vaccines against tuberculosis in cattle, goats, and other animal species have been the focus of research in this field during the last two decades. Improving the vaccine efficacy is essential prior to running long-lasting and expensive field trials. Studies have shown that vaccine protocols utilizing boosting with proteins improve the vaccine efficacy. The use of polymers such as chitosan and PolyLactic-co-Glycolic Acid (PLGA) improves the immune response against different diseases by improving the interaction of antigens with the cellular immune system and modulating the host immune response. This study shows that the prime BCG vaccination, boosted with a culture filtrate protein (CFP), alone or in combination with chitosan and PolyLactic-co-Glycolic Acid (PLGA), have the potential to reduce tuberculosis (TB) dissemination by reducing the number of animals with lesions, the number of lesions per animal, and the size of the lesions in vaccinated animals, compared with those not vaccinated or those vaccinated with BCG alone. The vaccinated groups showed significantly higher Interferon-γ levels in the blood compared to the control, nonvaccinated group after vaccination, after boosting, and after the challenge with the wild-type Mycobacterium bovis strain.

Molecular epidemiology of cattle tuberculosis in Mexico through whole-genome sequencing and spoligotyping.

Mycobacterium bovis infection in cattle persists in Mexico, posing a threat to human health. Control of bovine tuberculosis, through the National Program Against Bovine Tuberculosis, has led to the decrease of disease prevalence in most of the country, except for high dairy production regions. Genotyping of M. bovis has been performed mainly by spoligotyping and variable number tandem repeats (VNTR), but higher resolution power can be useful for a finer definition of the spread of the disease. Whole genome sequencing and spoligotyping was performed for a set of 322 M. bovis isolates from different sources in Mexico: Baja California, Coahuila, Estado de Mexico, Guanajuato, Hidalgo, Jalisco, Queretaro and Veracruz, from dairy and beef cattle, as well as humans. Twelve main genetic clades were obtained through WGS and genetic diversity analysis. A clear differentiation of the Baja California isolates was seen as they clustered together exclusively. However, isolates from the central states showed no specific clustering whatsoever. Although WGS proves to have higher resolving power than spoligotyping, and since there was concordance between WGS and spoligotyping results, we consider that the latter is still an efficient and practical method for monitoring bovine tuberculosis in developing countries, where resources for higher technology are scarce.

Characterization of Leptospira isolates from humans and the environment in Uruguay.

Laboratory diagnosis of human leptospirosis usually relies on indirect methods exploring specific immune response. Isolation and identification of the involved strains are cumbersome, but can provide biological resources for pathogenic studies and relevant information for guiding prevention and control measures. The aim of the research we are hereby reporting was the characterization of Leptospira isolates obtained from humans and the environment in Uruguay. Blood cultures were performed from early samples of 302 Uruguayan patients, mainly rural workers, and from 36 water samples taken from their living or working environments. Eight human isolates and seven environmental isolates were obtained and analyzed by end point Polymerase Chain Reaction (PCR), Multilocus Variable Number of Tandem Repeat Analysis (MLVA) and other molecular methods. Human isolates corresponded to several serogroups and serovars of Leptospira interrogans and Leptospira kirschneri species, probably reflecting the infection with similar involved Leptospira species and serovars of an extended animal reservoir in rural settings of the country, mostly dedicated to meat and dairy production. Culture-positive patients were older than usually affected workers, and presented signs and symptoms of severe illness. A high organic and circulating bacterial burden may explain an easier positive result from these workers' samples. Environmental isolates were mainly identified as Leptospira biflexa strains, with a single L. meyeri isolate of uncertain significance.

Genetic diversity based on MIRU-VNTR profile of isolates of Mycobacterium bovis from Mexican cattle.

Bovine tuberculosis (bTB) is a disease caused by Mycobacterium bovis (M. bovis), which affects cattle, animal species and humans. To determinate the genetic structure of strains of M. bovis in mexican cattle, 467 isolates obtained from 2009 to 2010 from different regions of Mexico with known spoligotype were included in the study. The isolates were genotyped by interspersed repeated mycobacterial units-variable number tandem repeats (MIRU-VNTR) obtaining 13 MIRU-VNTR groups. When combining MIRU-VNTR patterns with its spolygotypes, the Hunter genetic discrimination index (HGDI), we obtained 421 genetic patterns distributed in 17 groups. The HGDI for the total loci was 0.99. The locus that presented the higher HGDI was 2461 (0.857), while the locus with the lowest HGDI was 2686 (0.239). When we analyzed our results, using just 6 or 8 MIRU-VNTR we obtained an discriminatory power of 0.8499 and 0.8875 respectively indicating lower HGDI than 12 MIRU-VNTR locus.

Molecular Relationship between Strains of M. bovis from Mexico and Those from Countries with Free Trade of Cattle with Mexico.

The purpose of this study was to identify relationships between spoligotypes of M. bovis from cattle in Mexico and those reported in countries with free trade of cattle with Mexico: Australia, Canada, New Zealand and the United States of America. Mexican spoligotypes were obtained from isolates collected from cattle in different parts of the country. Spoligotypes from Canada and New Zealand were obtained from different reports in the literature. Those from the United States were obtained from the database of the National Veterinary Services Laboratory in APHIS-USDA. In order to perform the analysis in a single data set, spoligotypes were all converted to binary data and classified according to www.mbovis.org or www.pasteur-guadeloupe.fr:8081. Epidemiologic information included country and species infected. From 3,198 isolates, 174 different spoligotypes were obtained, 95 were orphans. Ninety one percent of the isolates came from the Unites States (n = 1,609) and Mexico (n = 1,323). Spoligotype SB0265 is shared between Canada and the United States in cattle and wildlife. Six spoligotypes, SB0673, SB0121, SB0145, SB0971, SB0140 and SB1165, were frequent in cattle and wildlife in the United States and cattle in Mexico, suggesting wide exchange of strains. Spoligotype SB0669 was found only in Mexico. Spoligotype SB0140 was the most common in Australia and the sixth in the United States and Mexico. In a phylogenetic analysis, spoligotype SB0140 appears as the oldest spoligotype in the data set, suggesting this as the ancestral spoligotype for all spoligotypes in the five countries. Some spoligotypes are shared by animals and humans, corroborating the zoonotic importance of M. bovis.

BmVDAC upregulation in the midgut of Rhipicephalus microplus, during infection with Babesia bigemina.

The molecular mechanisms involved during the infection of Rhipicephalus microplus midgut cells by Babesia bigemina are of great relevance and currently unknown. In a previous study, we found a voltage-dependent anion channel (VDAC)-like protein (BmVDAC) that may participate during parasite invasion of midgut cells. In this work, we investigated BmVDAC expression at both mRNA and protein levels and examined BmVDAC localization in midgut cells of ticks infected with B. bigemina at different times post-repletion. Based on the RT-PCR results, Bmvdac expression levels were significantly higher in infected ticks compared to uninfected ones, reaching their highest values at 24h post-repletion (p<0.0001). Similar results were obtained at the protein level (p<0.0001). Interestingly, BmVDAC immunolocalization showed that there was an important differential expression and redistribution of BmVDAC protein between the midgut cells of infected and uninfected ticks, which was more evident 24h post-repletion of infected ticks. This is the first report of BmVDAC upregulation and immunolocalization in R. microplus midgut cells during B. bigemina infection. Further studies regarding the function of BmVDAC during the infection may provide new insights into the molecular mechanisms between B. bigemina and its tick vector and could result in its use as an anti-tick and transmission-blocking vaccine candidate.

Valoración de un procedimiento de inmunofluorescencia indirecta para la detección de anticuerpos tipo IgM (IF-IgM) utilizado en el diagnóstico temprano de leptospirosis / Evaluation of an indirect immunofluorescence assay for detection of IgM (IF-IgM) type antibodies in the early diagnosis of leptospirosis

Introducción: la leptospirosis es una enfermedad febril, aguda, que presenta manifestaciones clínicas variadas. Esto dificulta o retarda el diagnóstico clínico, por lo cual es útil disponer de métodos de laboratorio adecuados para orientar el manejo inicial de estos pacientes. Objetivo: evaluar un procedimiento de inmunofluorescencia para detectar IgM (IF-IgM) de elaboración propia aplicado al diagnóstico temprano de leptospirosis. Material y método: se analizaron por IF-IgM y microaglutinación (MAT) (tomada como estándar de referencia) sueros obtenidos de pacientes con sospecha clínica de leptospirosis. La sensibilidad y especificidad de la IF-IgM versus MAT se establecieron utilizando una tabla de doble entrada. La concordancia entre dos observadores se determinó por el test Kappa. Resultados: de 161 muestras precoces analizadas, 97 sueros correspondieron a pacientes con infección aguda confirmada por MAT y 64 sin infección. La sensibilidad y especificidad de la IF-IgM con sueros de fase aguda fueron 79% y 100%, respectivamente. El índice Kappa fue 1. Conclusiones: la IF-IgM aparece como una herramienta útil para el diagnóstico temprano de pacientes con leptospirosis. No requiere el manejo de bacterias viables, puede realizarse en laboratorios que cuenten con microscopio de luz ultravioleta, se necesita una sola muestra de suero y el resultado está listo en tres a cuatro horas. En cuanto a las desventajas, no identifica los serovares involucrados y un resultado negativo no descarta la infección. Teniendo en cuenta esto último es obligatorio analizar por MAT una segunda muestra de suero obtenida a los 10-20 días de la primera para descartarla o confirmarla. (AU)

Introduction: leptospirosis is a severe febrile disease, with a variety of clinical presentations. This results in the delay of clinical diagnosis, or in difficulties achieving it, so having the appropriate laboratory tests may deem useful to guide the initial care of these patients. Objective: to evaluate a self-made immunofluorescence technique for the detection of IgM antibodies (IF-IgM) in the early diagnosis of leptospirosis. Method: serum samples obtained from patients with a clinical suspicion of leptospirosis were analysed by IF-IgM antibodies and the microagglutination test (MAT) . Sensitivity and specificity of IF-IgM versus MAT were determined using a double entry table. Agreement between two observers was determined by the Kappa test. Results: out of one hundred and sixty one early samples analysed, 97 serum samples corresponded to patients with a confirmed acute infection by MAT and 64 evidenced no infection. Sensitivity and specificity of IF-IgM assays in the acute phase were 79% and 100% respectively. The Kappa value was 1.Conclusions: IF-IgM seems to be a useful tool for the early diagnosis of patients with leptospirosis. It does not need viable bacteria to be handled; it may be applied in laboratories equipped with ultraviolet light microscopes, a single serum sample is needed and the result is seen three to four hours later. As to its disadvantages, it fails to identify the involved serovars and a negative result does not discard infection. Bearing the latter into account, the MAT test is mandatory in a second serum sample obtained 10-20 days after the first one, to either discard or confirm the disease.

Introdução: a leptospirose é uma doença febril, aguda, com diferentes manifestações clínicas o que dificulta ou atrasa o diagnóstico clínico. Por essa razão é importante contar com métodos de laboratório adequados para orientar o manejo inicial dos pacientes.Objetivo: avaliar uma técnica de imunofluorescencia para detectar IgM (IF-IgM) para o diagnóstico precoce de leptospirose.Material e método: foram analisados por IF-IgM e microaglutinaçao (MAT) (usada como padrão de referencia) soros de pacientes com suspeita clínica de leptospirose. A sensibilidade e a especificidade da IF-IgM comparada com MAT foi determinada usando uma tabela de dupla entrada. A concordância entre dois observadores foi determinada usando o teste Kappa.Resultados: Cento sessenta e uma amostras da fase precoce foram analisadas, sendo 97 de pacientes com infecção aguda confirmada por MAT e 64 sem infecção. A sensibilidade e a especificidade da IF-IgM com soros obtidos na fase aguda foram 79% e 100%, respectivamente. O índice Kappa foi igual a 1.Conclusões: a técnica IF-IgM que desenvolvemos, pode ser uma ferramenta útil para o diagnóstico precoce de pacientes com leptospirose. Não é necessário manipular bactérias viáveis, pode ser realizada em laboratórios que tenham microscópio com luz ultravioleta, somente é necessária uma amostra de soro e o resultado está disponível em três-quatro horas. Com relação às desvantagens, esta técnica não identifica os serovares e um resultado negativo não descarta a presença de infecção. Considerando estas desvantagens, é obrigatório analisar por MAT uma segunda amostra de soro obtida 10 a 20 dias depois da primeira para descartar ou confirmar a infecção.

Valoración de un procedimiento de inmunofluorescencia indirecta para la detección de anticuerpos tipo IgM (IF-IgM) utilizado en el diagnóstico temprano de leptospirosis / Evaluation of an indirect immunofluorescence assay for detection of IgM (IF-IgM) type antibodies in the early diagnosis of leptospirosis

Introducción: la leptospirosis es una enfermedad febril, aguda, que presenta manifestaciones clínicas variadas. Esto dificulta o retarda el diagnóstico clínico, por lo cual es útil disponer de métodos de laboratorio adecuados para orientar el manejo inicial de estos pacientes. Objetivo: evaluar un procedimiento de inmunofluorescencia para detectar IgM (IF-IgM) de elaboración propia aplicado al diagnóstico temprano de leptospirosis. Material y método: se analizaron por IF-IgM y microaglutinación (MAT) (tomada como estándar de referencia) sueros obtenidos de pacientes con sospecha clínica de leptospirosis. La sensibilidad y especificidad de la IF-IgM versus MAT se establecieron utilizando una tabla de doble entrada. La concordancia entre dos observadores se determinó por el test Kappa. Resultados: de 161 muestras precoces analizadas, 97 sueros correspondieron a pacientes con infección aguda confirmada por MAT y 64 sin infección. La sensibilidad y especificidad de la IF-IgM con sueros de fase aguda fueron 79% y 100%, respectivamente. El índice Kappa fue 1. Conclusiones: la IF-IgM aparece como una herramienta útil para el diagnóstico temprano de pacientes con leptospirosis. No requiere el manejo de bacterias viables, puede realizarse en laboratorios que cuenten con microscopio de luz ultravioleta, se necesita una sola muestra de suero y el resultado está listo en tres a cuatro horas. En cuanto a las desventajas, no identifica los serovares involucrados y un resultado negativo no descarta la infección. Teniendo en cuenta esto último es obligatorio analizar por MAT una segunda muestra de suero obtenida a los 10-20 días de la primera para descartarla o confirmarla...

Usefulness of real-time PCR assay targeting lipL32 gene for diagnosis of human leptospirosis in Uruguay.

INTRODUCTION: Assays based on DNA amplification can provide information that contributes to the initial management of patients with leptospirosis. However, these have not been adopted in Uruguay. Our aim was to evaluate the performance of the lipL32 real-time PCR (qPCR) for diagnosis of leptospirosis. METHODOLOGY: We analyzed by microscopic agglutination test (MAT) and lipL32 qPCR serum samples from 183 patients with suspected leptospirosis. To establish the analytical sensitivity of the qPCR, experimentally spiked samples with known amounts of Leptospira interrogans were analyzed. RESULTS: The analytical sensitivity of the qPCR was 102 leptospires/mL. In 98 patients MAT results were negative meanwhile 85 showed positive reactions, revealing acute infections. Twenty six acute-phase sera of these 85 patients showed a positive signal by qPCR (diagnostic sensitivity 30%). In these patients the average time between onset of symptoms and collection of the first sample was 8 days. In patients with negative results for qPCR and positive MAT results (n=59) the average interval between onset of symptoms and collection of the first sample was 13 days. The qPCR did not yield false positive results. CONCLUSIONS: The qPCR had a lower diagnostic sensitivity than MAT and a higher cost. However, it allowed to make an early diagnosis in 26 patients. In patients with confirmed acute infections and negative results by qPCR, more than 8 days had elapsed between the onset of the illness and extraction of the first serum sample. Our data support that the qPCR from sera have clinical utility within the first week of illness.

A ten-year follow-up of human leptospirosis in Uruguay: an unresolved health problem.

Leptospira spp. are delicate bacteria that cannot be studied by usual microbiological methods. They cause leptospirosis, a zoonotic disease transmitted to humans through infected urine of wild or domestic animals. We studied the incidence of this disease in the Uruguayan population, its epidemiologic and clinical features, and compared diagnostic techniques. After examining 6,778 suspect cases, we estimated that about 15 infections/100,000 inhabitants occurred yearly, affecting mainly young male rural workers. Awareness about leptospirosis has grown among health professionals, and its lethality has consequently decreased. Bovine infections were probably the principal source of human disease. Rainfall volumes and floods were major factors of varying incidence. Most patients had fever, asthenia, myalgias or cephalalgia, with at least one additional abnormal clinical feature. 30-40% of confirmed cases presented abdominal signs and symptoms, conjunctival suffusion and altered renal or urinary function. Jaundice was more frequent in patients aged > 40 years. Clinical infections followed an acute pattern and their usual outcome was complete recovery. Laboratory diagnosis was based on indirect micro-agglutination standard technique (MAT). Second serum samples were difficult to obtain, often impairing completion of diagnosis. Immunofluorescence was useful as a screening test and for early detection of probable infections.

A ten-year follow-up of human leptospirosis in Uruguay: an unresolved health problem / Diez años de seguimiento de la leptospirosis humana en Uruguay: un problema de salud no resuelto

Leptospira spp. are delicate bacteria that cannot be studied by usual microbiological methods. They cause leptospirosis, a zoonotic disease transmitted to humans through infected urine of wild or domestic animals. We studied the incidence of this disease in the Uruguayan population, its epidemiologic and clinical features, and compared diagnostic techniques. After examining 6,778 suspect cases, we estimated that about 15 infections/100,000 inhabitants occurred yearly, affecting mainly young male rural workers. Awareness about leptospirosis has grown among health professionals, and its lethality has consequently decreased. Bovine infections were probably the principal source of human disease. Rainfall volumes and floods were major factors of varying incidence. Most patients had fever, asthenia, myalgias or cephalalgia, with at least one additional abnormal clinical feature. 30-40% of confirmed cases presented abdominal signs and symptoms, conjunctival suffusion and altered renal or urinary function. Jaundice was more frequent in patients aged > 40 years. Clinical infections followed an acute pattern and their usual outcome was complete recovery. Laboratory diagnosis was based on indirect micro-agglutination standard technique (MAT). Second serum samples were difficult to obtain, often impairing completion of diagnosis. Immunofluorescence was useful as a screening test and for early detection of probable infections.

El género Leptospira comprende bacterias delicadas, que requieren métodos de estudio especiales. Causan una zoonosis transmitida a los seres humanos por la orina de animales domésticos o silvestres. Estudiamos la incidencia de la leptospirosis en la población humana de Uruguay, sus características epidemiológicas y clínicas, y comparamos técnicas diagnósticas. Tras examinar 6778 casos sospechosos, estimamos que anualmente ocurren unas 15 infecciones/100.000 habitantes, principalmente en trabajadores rurales jóvenes de sexo masculino. El nivel de alerta sobre la leptospirosis ha avanzado en el personal sanitario, y su letalidad ha descendido. La enfermedad humana se asocia principalmente con las infecciones bovinas. Lluvias e inundaciones influyen sobre su incidencia. La mayoría de los pacientes experimentaron fiebre, astenia, mialgias o cefaleas, con al menos un síntoma o signo adicional. 30 a 40% presentaban sintomatología abdominal, hiperemia conjuntival o alteraciones nefro-urológicas. La ictericia era más frecuente en mayores de 40 años. La enfermedad era habitualmente aguda y la recuperación completa. El diagnóstico de laboratorio se basó en la técnica de microaglutinación (MAT), con cepas vivas de distintos serovares. Se requieren segundas muestras de suero, de difícil obtención. El test de inmunofluorescencia se reveló útil para screening y detección precoz de la infección.

Prevalencia de polimorfismos del receptor de vitamina D en pacientes pediátricos mexicanos con enfermedad renal crónica / Prevalence of vitamin D receptor gene polymorphisms in Mexican children with chronic kidney disease

Introducción. La posible influencia de los diferentes polimorfismos del receptor de vitamina D sobre la regulación del eje calcio-hormona paratiroidea -vitamina D es particularmente importante en pacientes con enfermedad renal crónica, ya que cursan con trastornos en el metabolismo mineral óseo. El objetivo del trabajo fue describir la frecuencia de los polimorfismos rs7975232 (ApaI), rs731236 (TaqI) y rs1544410 (BsmI) en el receptor de vitamina D y su relación con el calcio, el fósforo, la fosfatasa alcalina y la hormona paratiroidea en pacientes pediátricos con enfermedad renal crónica. Métodos. Se incluyeron 31 pacientes con enfermedad renal crónica. Se tomaron muestras de sangre a cada uno para la determinación de marcadores bioquímicos y muestras de ADN para la genotipificación del gen del receptor a vitamina D utilizando el método de la reacción en cadena de la polimerasa y restricción enzimática. Resultados. La edad promedio de los pacientes fue de 15.6 ± 3.6 años. Para ApaI, fueron homocigotos AA siete pacientes (22.58%); Aa, 18 pacientes (58.06%) y AA, seis pacientes (19.35%). Para BsmI, 14 pacientes fueron BB (45.16%) y 17 pacientes fueron Bb (54.83%). Los pacientes BB tuvieron calcio sérico mayor que los Bb (p = 0.04). Para TaqI, fueron TT 19 pacientes (61.3%); Tt, once pacientes (35.48%) y uno, tt (3.22%). Aquellos con alelo t (Tt o tt) presentaron concentraciones de calcio sérico significativamente más bajas que el homocigoto TT. Conclusiones. Se reportaron las frecuencias de genotipos y alelos del receptor de vitamina D en niños mexicanos. Se obtuvieron valores de calcio sérico más elevados en aquellos con alelo BB o TT.

Background. The possible influence of vitamin D receptor (VDR) gene polymorphisms on the regulation of the calcium-parathyroid hormone-vitamin D axis is particularly important in end-stage renal disease (ESRD) patients due to alterations in bone mineral metabolism. The aim of the study was to describe the frequency of VDR gene polymorphisms rs7975232 (ApaI), rs731236 (TaqI), rs1544410 (BsmI) and their relationship with serum calcium, phosphate, alkaline phosphatase and parathyroid hormone in children with ESRD. Methods. Thirty one patients were included in the study. Blood samples were drawn for biochemical determinations and DNA extraction. VDR genotypes were analyzed as restriction fragment length polymorphisms using ApaI, BsmI and TaqI. Results. Mean age of the patients was 15.6 ± 3.6 years. For ApaI, seven patients were AA (22.58%), 18 patients were Aa (58.06%) and six patients were aa (19.35%). For BsmI, 14 patients were BB (45.16%) and 17 were Bb (54.83%). Patients who were homozygous for BB had higher serum calcium concentrations than Bb patients (p = 0.04). For TaqI, 19 patients were TT (61.3%), 11 were Tt (35.48%) and one was tt (3.22%). Those with t allele (Tt or tt) had serum calcium concentrations significantly lower than TT patients. Conclusions. Genotype and allele frequency of the VDR gene is reported in Mexican patients with ESRD. Those with BB or TT genotype had higher serum calcium concentrations.

The identification of a VDAC-like protein involved in the interaction of Babesia bigemina sexual stages with Rhipicephalus microplus midgut cells.

We investigated the interaction of Rhipicephalus microplus midgut cells with Babesia bigemina sexual stages using a proteomic approach. A polypeptide from the R. microplus midgut that binds to proteins from B. bigemina sexual stages was identified and sequenced. Combining 2D overlay and tandem mass spectrometry (MS/MS) techniques, we determined that this polypeptide corresponds to a mitochondrial voltage-dependent anion-selective channel (VDAC). The vdac gene encoding the sequenced polypeptide was identified and sequenced. This is the first report of a VDAC-like protein in R. microplus, and a possible role for this protein in the B. bigemina infection process is suggested.

Falla de filtros para leucocitos para prevenir infección por citomegalovirus en un paciente con trasplante de médula ósea / Leukocyte-filter failure to prevent cytomegalovirus transmission in a patient with bone marrow transplantation

Introducción. La infección por citomegalovirus (CMV) adquirida mediante productos sanguÌneos puede llegar a ser importante causa de morbilidad y mortalidad en aquellos pacientes sometidos a trasplante de médula ósea (TMO). En vista de la dificultad para contar con donadores CMV negativos se utilizan filtros para leucocitos en los productos transfundidos, con reportes de efectividad de hasta 99 por ciento en pacientes CMV negativos.Caso clínico. Paciente masculino de 12 años de edad con diagnósticos de leucemia aguda no linfoblástica M4, CMV negativo por serología y PCR, que recibió TMO autólogo con purga con mafosfamida. Los productos sanguíneos se filtraron para leucocitos. Injerto de médula ósea el día +14, y el día +28 presentó disminución de leucocitos a 200/µL. La PCR fue positiva para CMV en orina, plasma y leucocitos; inició tratamiento con ganciclovir con lo cual la cifra de leucocitos se incrementó paulatinamente después de negativizar la PCR. Conclusión. Ya que la población seropositiva para CMV es muy alta en nuestro medio, es difícil obtener productos seronegativos, y a pesar del uso de filtros para leucocitos existe riesgo de transmitir el virus como es el caso que se presenta. Se considera que se requiere un escrutinio en banco de sangre para detectar donadores con infección reciente y viremia.