Association Study of CACNA1D, KCNJ11, KCNQ1, and CACNA1E Single-Nucleotide Polymorphisms with Type 2 Diabetes Mellitus.

Type 2 diabetes mellitus (T2DM) is a complex chronic disease characterized by decreased insulin secretion and the development of insulin resistance. Previous genome-wide association studies demonstrated that single-nucleotide polymorphisms (SNPs) present in genes coding for ion channels involved in insulin secretion increase the risk of developing this disease. We determined the association of 16 SNPs found in CACNA1D, KCNQ1, KCNJ11, and CACNA1E genes and the increased probability of developing T2DM. In this work, we performed a case-control study in 301 Mexican adults, including 201 cases with diabetes and 100 controls without diabetes. Our findings indicate a moderate association between T2DM and the C allele, and the C/C genotype of rs312480 within CACNA1D. The CAG haplotype surprisingly showed a protective effect, whereas the CAC and CGG haplotypes have a strong association with T2DM. The C allele and C/C genotype of rs5219 were significantly associated with diabetes. Also, an association was observed between diabetes and the A allele and the A/A genotype of rs3753737 and rs175338 in CACNA1E. The TGG and CGA haplotypes were also found to be significantly associated. The findings of this study indicate that the SNPs examined could serve as a potential diagnostic tool and contribute to the susceptibility of the Mexican population to this disease.

Genetic Variation among the Partial Gene Sequences of the Ribosomal Protein Large-Two, the Internal Transcribed Spacer, and the Small Ribosomal Subunit of Blastocystis sp. from Human Fecal Samples.

In the present study, we compared the genetic variability of fragments from the internal transcribed spacer region (ITS) and the small subunit ribosomal DNA (SSUrDNA) as nuclear markers, in contrast with the ribosomal protein large two (rpl2) loci, placed in the mitochondrion-related organelles (MROs) within and among human fecal samples with Blastocystis. Samples were analyzed using polymerase chain reaction (PCR)-sequencing, phylogenies, and genetics of population structure analyses were performed. In total, 96 sequences were analyzed, i.e., 33 of SSUrDNA, 35 of rpl2, and 28 of ITS. Only three subtypes (STs) were identified, i.e., ST1 (11.4%), ST2 (28.6%), and ST3 (60%); in all cases, kappa indexes were 1, meaning a perfect agreement among ST assignations. The topologies of phylogenetic inferences were similar among them, clustering to each ST in its specific cluster; discrepancies between phylogeny and assignment of STs were not observed. The STRUCTURE v2.3.4 software assigned three subpopulations corresponding to the STs 1-3, respectively. The population indices were consistent with those previously reported by other groups. Our results suggest the potential use of the ITS and rpl2 genes as molecular markers for Blastocystis subtyping as an alternative approach for the study of the genetic diversity observed within and between human isolates of this microorganism.

Identification of SARS-CoV-2 in urban rodents from Southern Mexico City at the beginning of the COVID-19 pandemic.

Currently, there are some concerns about the situation and, in particular, about the future of the COVID-19 pandemic and the new emerging variants of SARS-CoV-2. Rodents are an example of synanthropic animals in urban environments that harbor important zoonoses. Although the molecular identification of SARS-CoV-2 in Rattus norvegicus from New York City had been reported, in other studies, urban wild rodents infected with this virus have not been found. This study aimed to molecularly identify the presence of SARS-CoV-2 in urban wild rodents from Mexico City, trapped along a water channel of a public park as part of a pest control program, at the beginning of the COVID-19 pandemic, during the fall and winter of 2020. Up to 33 Mus musculus and 52 R. norvegicus were captured and euthanized, large intestine samples with feces from the animals were obtained. RNAs were obtained and subjected to qRT-PCR for SARS-CoV-2 identification and threshold cycle (Ct) values were obtained. Four mice (12.1%) and three rats (5.8%) were positive, three rodents exhibited Ct<30. Our results on the frequency of SARS-CoV-2 in urban rats are in line with other previous reports. Thus, similar to other authors, we suggest that surveillance for the detection of SARS-CoV-2 in urban wild rodents, as sentinel animals, should be maintained.

Identification of SARS-CoV-2 in urban rodents from Southern Mexico City at the beginning of the COVID-19 pandemic

ABSTRACT Currently, there are some concerns about the situation and, in particular, about the future of the COVID-19 pandemic and the new emerging variants of SARS-CoV-2. Rodents are an example of synanthropic animals in urban environments that harbor important zoonoses. Although the molecular identification of SARS-CoV-2 in Rattus norvegicus from New York City had been reported, in other studies, urban wild rodents infected with this virus have not been found. This study aimed to molecularly identify the presence of SARS-CoV-2 in urban wild rodents from Mexico City, trapped along a water channel of a public park as part of a pest control program, at the beginning of the COVID-19 pandemic, during the fall and winter of 2020. Up to 33 Mus musculus and 52 R. norvegicus were captured and euthanized, large intestine samples with feces from the animals were obtained. RNAs were obtained and subjected to qRT-PCR for SARS-CoV-2 identification and threshold cycle (Ct) values were obtained. Four mice (12.1%) and three rats (5.8%) were positive, three rodents exhibited Ct<30. Our results on the frequency of SARS-CoV-2 in urban rats are in line with other previous reports. Thus, similar to other authors, we suggest that surveillance for the detection of SARS-CoV-2 in urban wild rodents, as sentinel animals, should be maintained.

Salud sexual de mujeres universitarias de guanajuato, méxico

Objetivo: Describir la salud sexual global y por dimensión de un grupo de mujeres universitarias y determinar su relación con variables sociodemográficas, académicas y caracterización en sexualidad. Material y Método: Investigación cuantitativa, descriptiva, correlacional, realizada en 91 mujeres de dos universidades privadas de Guanajuato, se aplicó una cédula de datos y la Escala de salud sexual; el muestreo para las universidades fue probabilístico y para las participantes no probabilístico; se utilizó estadística descriptiva e inferencial y se apegó a las consideraciones ético-legales. Resultados: Participaron 91 mujeres de 18 a 24 años, 74,7% con vida sexual, refirieron de 1 a 11 parejas sexuales, una media de 17 años (DE= 1,73) para inicio de vida sexual, obtuvieron niveles altos de salud sexual (x= 114,85; DE= 11,50) y se encontró relación estadísticamente significativa en variables como inicio de vida sexual, edad de inicio de vida sexual, número de parejas sexuales, vida sexual activa y uso de métodos anticonceptivos. Conclusiones: Las participantes cuentan con niveles altos de salud sexual, así como de conocimientos y actitudes, sin embargo, presentan un nivel medio de prácticas, situación que vulnera la salud y otros aspectos. Este estudio contribuye a brindar un panorama sobre la salud sexual de las jóvenes, así como algunas variables que enfermería debe considerar para la formulación de programas que coadyuven a fomentar la salud sexual.

Objective: To describe the overall sexual health, as well as specific dimensions, of a group of university women and to determine its relationship with sociodemographic, academic and sexuality characterizing variables. Materials and Methods: Quantitative, descriptive, correlational study, carried out on 91 women from two private universities in Guanajuato (Mexico), using a data sheet and the Sexual Health Scale, with probability sampling for the universities and non-probability sampling for the participants. Descriptive and inferential statistics were used and ethical-legal considerations were followed. Results: 91 women aged 18 to 24 years participated, 74.7% had an active sexual life, reported from one to 11 sexual partners, a mean age of 17 years (SD 1.73) for sexual initiation, showed high levels of sexual health awareness (X114.85; SD 11.50) and a statistically significant relationship was found in variables such as sexual initiation, age at sexual initiation, number of sexual partners, sexually active life and use of contraceptive methods. Conclusions: The participants presented a high level of sexual health awareness, as well as knowledge and attitudes, but they presented a medium level of practices, a situation that is detrimental to health and other aspects. This study aims to provide an overview of the sexual health of young women, as well as some variables that nursing practice should consider when formulating programs that help promote sexual health.

Objetivo: Descrever a saúde sexual geral, e por dimensão específica, de um grupo de mulheres universitárias e determinar sua relação com variáveis sociodemográficas, académicas e de caracterização da sexualidade. Material e métodos: Pesquisa quantitativa, descritiva, correlacional, realizada com 91 mulheres de duas universidades privadas de Guanajuato (México), usando uma ficha técnica e a Escala de Saúde Sexual, com amostragem probabilística para as universidades e amostragem não probabilística para as participantes. Foram usadas estatísticas descritivas e inferenciais e respeitaram-se os aspectos ético-legais. Resultados: Participaram 91 mulheres de 18 a 24 anos, 74,7% tinham vida sexual ativa, relataram de um a 11 parceiros sexuais, uma idade média de 17 anos (DP 1,73) para o início da vida sexual, apresentaram altos níveis de saúde sexual (x 114,85; DP 11,50) e foi encontrada uma relação estatisticamente significativa em variáveis como início da vida sexual, idade de início da vida sexual, número de parceiros sexuais, vida sexual ativa e uso de métodos contraceptivos. Conclusões: Os participantes apresentaram elevados níveis de saúde sexual, bem como conhecimentos e atitudes, porém apresentaram um nível médio de práticas, situação que prejudica a saúde e outros aspectos. Este estudo tem como objetivo fornecer um panorama sobre a saúde sexual de mulheres jovens, bem como algumas variáveis que a prática de enfermagem deve considerar na formulação de programas que ajudem a promover a saúde sexual.

Presence of human adenovirus 36 in visceral fat tissue, viral load, and analysis of its genetic variability.

It has been proposed that infection by adipogenic viruses constitutes a "low risk" factor for obesity. Here, we report the presence of adenovirus 36 (Ad36) and its viral load copy number in fat tissue of participants with obesity and normal weight; phylogenetic analysis was performed to describe their relationship and genetic variability among viral haplotypes. Adipose tissue obtained from 105 adult patients with obesity (cases) and 26 normal-weight adult participants as controls were analyzed by quantitative polymerase chain reaction (qPCR) amplifying the partial Ad36 E1a gene. The amplicons were examined by melting curves and submitted to sequencing. Then, genetic diversity and phylogenetic inferences were performed. Ad36 was identified at rates of 82% and 46% in the case and control groups, respectively (p = 1.1 × 10-4 , odds ratio = 5.28); viral load copies were also significantly different between both groups, being 25% higher in the case group. Melting curve analysis showed clear amplification among positive samples. Phylogenetic inferences and genetic diversity analyses showed that the Ad36 E1a gene exhibits low genetic variability and differentiation with strong gene flow due to an expanding process. Our results suggest that the phenomenon of infectobesity by Ad36 might not be a low-risk factor, as has been previously argued by other authors.

Identification of Blastocystis sp. in school children from a rural Mexican village: subtypes and risk factors analysis.

Blastocystis sp. is a common eukaryotic microorganism that colonizes the intestinal tract of several animals, including humans, although its role as a pathogen is still unclear. In the present study, we report the prevalence and risk factors associated with Blastocystis infection in scholars from a rural community in Mexico. A cross-sectional observational study was carried out on schoolchildren aged 3 to 15 years old; fecal samples were analyzed by culture, Faust technique, and molecular analysis. In addition, a structured questionnaire was applied to identify possible risk factors. Of the 177 samples obtained, Blastocystis sp. was the microorganism that presented the highest frequency (n=78, 44%), and included the following subtypes (STs): ST1 (n=43, 56.5%), ST2 (n=18, 23.6%), and ST3 (n=15, 19.7%); Blastocystis STs were not identified in two cases. No associating factors were found between Blastocystis infection or among STs vs. symptoms. During bivariate analysis, no statistically significant risk factors were found, except for the variable of "eating sweets, snacks, and handmade food on the way home" (p=0.04). Therefore, it is plausible to conclude that schoolchildren become infected with Blastocystis sp. mainly outside their homes, perhaps by eating contaminated handmade food on their way to or from school; however, this variable should be evaluated in detail in future studies.

Factores relacionados a la depresión durante la adolescencia: una revisión integrativa / Factors related to depression during adolescence: an integrative review

INTRODUCCIÓN: La depresión es un problema de salud mental común en la etapa de la adolescencia, se manifiesta por descenso del humor, tristeza y pérdida de interés en actividades cotidianas. Esta etapa es sensible por los grandes cambios biopsicosociales. OBJETIVO: identificar factores relacionados a la depresión en adolescentes que puedan actuar como factores protectores o factores de riesgo. METODOLOGÍA: se realizó una búsqueda bibliográfica en las bases de datos WoS, PUBMED, Scopus, y BVS; se utilizaron descriptores normalizados para la expresión de búsqueda "Adolescente AND factores protectores OR factores de riesgo AND depresión", seleccionando 38 artículos. RESULTADOS: se obtuvieron 34 factores, que pueden actuar como de riesgo y protectores, y agrupados en dimensiones: a) biológica: género, edad, índice de masa corporal, problemas de salud; b) psicológica: autorregulación, autoestima, afecto positivo/negativo, pensamientos negativos, imagen corporal, estrés, alexitimia, calidad de vida, y c) social, subdividida en tres grupos: c.1) hábitos: consumo de sustancias nocivas, actividad física/sedentarismo, adicción a pantallas, rendimiento académico, participación comunitaria, estilo de vida, actividad sexual, sueño; c.2) contexto familiar: experiencias familiares, relación padres-hijos, funcionalidad familiar, composición familiar, nivel socioeconómico; y c.3) entorno: escuela urbana, implicación escolar, bullying, apoyo social, exposición a violencia, eventos vitales negativos, alfabetización en salud y áreas verdes. CONCLUSIÓN: Existen factores relacionados a la depresión en adolescentes que podrán actuar como factores protectores o de riesgo, su conocimiento por parte de los profesionales de la salud y de la enfermera en particular es fundamental para intervenirlos.

INTRODUCTION: Depression is a common mental health problem in adolescence, manifested by poor mood, sadness and loss of interest in daily activities. Adolescents are especially susceptible to depression due to the great biopsychosocial changes in this stage of life. OBJECTIVE: To identify risk factors and protective factors associated with adolescent depression that are evidence-based. METHODOLOGY: a bibliographic search was carried out in the WoS, PUBMED, Scopus, and VHL databases. Standardized descriptors used to conduct the search included Adolescent AND protective factors OR risk factors AND depression. 38 articles were selected. RESULTS: 38 factors were identified, classified as risky and protective, and grouped into the following dimensions: a) biological: gender, age, BMI, health problems; b) psychological: negative or positive affection, negative thoughts, satisfaction with body image, stress, alexhythemia, quality of life, self-regulation, self-esteem; and c) social, subdivided into three groups: c.1) habits, physical activity, consumption of harmful substances, screen addiction, lifestyle that needs to be improved, sexual activity, community participation, sleep duration, academic performance; c.2) family context: experiences, parent-child relationship, composition, socioeconomic level, functionality, educational level of parents; and c.3) environmental:: social support, bullying, exposure to violence, belonging to an urban school, negative life events, school involvement, neighborhood with green areas and health literacy. CONCLUSION: Several factors that affect depression in adolescents are reported by the literature. In the biological dimension, they tend to be risk factors, and in the psychological and social dimensions, they may increase risk or be protective. Knowledge of these factors by the nurse is essential to guide interventions.

Long-term infection passaging of Human Adenovirus 36 in monkey kidney cells.

Human Adenovirus 36 (HAdV-36) has been related to diverse effects on metabolism and may attenuate the lipid accumulation in kidneys with increased adiposity. Some of these effects would be related to viral persistence. However, until now, a model of persistent in vitro infection by HAdV-36 is unknown. In this study, we examined the cells of the Vero lineage to explore their permissiveness to long-term HAdV-36 infection. HAdV-36 was productively replicated in Vero cells and maintained long-term infection for up to 35 cell passages. A subculture was obtained from the cells that survived the primary infection at a low MOI (0.5). The production of the extracellular infectious virus with titers ranging from 104 to 106 TCID50/mL and DNA-bearing cells was detected. In long-term infected cells, the intracellular distribution of viral antigen was demonstrated by performing immunolocalization (IFI) and expression of cell-viral antigen in 50% of cells by flow cytometry, using anti-HAdV-36 hyperimmune rabbit serum. Furthermore, E1a and E4orf1 genes in long-term infected passages showed a decreasing trend. Our preliminary results reveal that renal epithelial monkey cells are permissive for the productive infection of HAdV-36. Vero cell culture long-term infection might be a promising model for addressing the fundamental aspects of the HAdV-36 biology that cannot reveal broadly-used cultures, which do not maintain long-term infection in primary or transformed cells.

Identification of α-L-fucosidase (ALFuc) of Blastocystis sp. subtypes ST1, ST2 and ST3.

Blastocystis sp. is a common intestinal microorganism. The α-L-fucosidase (ALFuc) is an enzyme long associated with the colonization of the gut microbiota. However, this enzyme has not been experimentally identified in Blastocystis cultures. The objective of the present study was to identify ALFuc in supernatants of axenic cultures of Blastocystis subtype (ST)1 ATCC-50177 and ATCC-50610 and to compare predicted ALFuc proteins of alfuc genes in sequenced STs1-3 isolates in human Blastocystis carriers. Excretion/secretion (Es/p) and cell lysate proteins were obtained by processing Blastocystis ATCC cultures and submitting them to SDS-PAGE and immunoblotting. In addition, 18 fecal samples from symptomatic Blastocystis human carriers were analyzed by sequencing of amplification products for subtyping. A complete identification of the alfuc gene and phylogenetic analysis were performed. Immunoblotting showed that the amplified band corresponding to ALFuc (~51 kDa) was recognized only in the ES/p. Furthermore, prediction analysis of ALFuc 3D structures revealed that the domain α-L-fucosidase and the GH29 family's catalytic sites were conserved; interestingly, the galactose-binding domain was recognized only in ST1 and ST2. The phylogenetic inferences of ALFuc showed that STs1-3 were clearly identifiable and grouped into specific clusters. Our results show, for the first time through experimental data that ALFuc is a secretion product of Blastocystis sp., which could have a relevant role during intestinal colonization; however, further studies are required to clarify this condition. Furthermore, the alfuc gene is a promising candidate for a phylogenetic marker, as it shows a conserved classification with the SSU-rDNA gene.

Sonographic cervical length predicts vaginal delivery after previous cesarean section in women with low Bishop score induced with a double-balloon catheter.

OBJECTIVE: To assess the role of cervical length when predicting vaginal delivery after a previous cesarean section (CS) in women with low Bishop score following the use of a double-balloon catheter for induction of labor (IOL). METHODS: A prospective, longitudinal study was conducted at a large teaching hospital in Santiago to recruit pregnant women at term with a previous CS and Bishop score ≤6 for IOL with a double-balloon catheter. The device was maintained for up to 24 h and the patient continued IOL with oxytocin only if the Bishop score was >6. Demographic and clinical variables were recorded and compared against vaginal delivery as the primary outcome. Multivariate logistic regression analysis was used to compare perinatal demographic and clinical variables in women achieving vaginal delivery versus those having a repeat CS. RESULTS: The final cohort included 40 pregnant women. Women achieving vaginal delivery (n = 17, 42.5%) had statistically significant differences in mean cervical length (24.8 mm versus 33.4 mm, respectively; p = .006), median Bishop score after removing the double-balloon catheter (11 versus 7, respectively; p = .005), and mean interval between double-balloon catheter placement and vaginal delivery or the decision to perform a CS (17.4 h versus 23.6 h, respectively; p = .03). Backward stepwise selection revealed an odds ratio of 0.90 (95% confidence interval = 0.82-0.98) for cervical length and a receiver operating characteristic curve area of 0.73. CONCLUSION: Cervical length, as determined by transvaginal sonography, proved to be effective in predicting vaginal delivery in women with a previous CS and low Bishop score following the use of a double-balloon catheter for IOL.

Identification of α-L-fucosidase (ALFuc) of Blastocystis sp. subtypes ST1, ST2 and ST3

ABSTRACT Blastocystis sp. is a common intestinal microorganism. The α-L-fucosidase (ALFuc) is an enzyme long associated with the colonization of the gut microbiota. However, this enzyme has not been experimentally identified in Blastocystis cultures. The objective of the present study was to identify ALFuc in supernatants of axenic cultures of Blastocystis subtype (ST)1 ATCC-50177 and ATCC-50610 and to compare predicted ALFuc proteins of alfuc genes in sequenced STs1-3 isolates in human Blastocystis carriers. Excretion/secretion (Es/p) and cell lysate proteins were obtained by processing Blastocystis ATCC cultures and submitting them to SDS-PAGE and immunoblotting. In addition, 18 fecal samples from symptomatic Blastocystis human carriers were analyzed by sequencing of amplification products for subtyping. A complete identification of the alfuc gene and phylogenetic analysis were performed. Immunoblotting showed that the amplified band corresponding to ALFuc (~51 kDa) was recognized only in the ES/p. Furthermore, prediction analysis of ALFuc 3D structures revealed that the domain α-L-fucosidase and the GH29 family's catalytic sites were conserved; interestingly, the galactose-binding domain was recognized only in ST1 and ST2. The phylogenetic inferences of ALFuc showed that STs1-3 were clearly identifiable and grouped into specific clusters. Our results show, for the first time through experimental data that ALFuc is a secretion product of Blastocystis sp., which could have a relevant role during intestinal colonization; however, further studies are required to clarify this condition. Furthermore, the alfuc gene is a promising candidate for a phylogenetic marker, as it shows a conserved classification with the SSU-rDNA gene.

Long-term infection passaging of Human Adenovirus 36 in monkey kidney cells

ABSTRACT Human Adenovirus 36 (HAdV-36) has been related to diverse effects on metabolism and may attenuate the lipid accumulation in kidneys with increased adiposity. Some of these effects would be related to viral persistence. However, until now, a model of persistent in vitro infection by HAdV-36 is unknown. In this study, we examined the cells of the Vero lineage to explore their permissiveness to long-term HAdV-36 infection. HAdV-36 was productively replicated in Vero cells and maintained long-term infection for up to 35 cell passages. A subculture was obtained from the cells that survived the primary infection at a low MOI (0.5). The production of the extracellular infectious virus with titers ranging from 104 to 106 TCID50/mL and DNA-bearing cells was detected. In long-term infected cells, the intracellular distribution of viral antigen was demonstrated by performing immunolocalization (IFI) and expression of cell-viral antigen in 50% of cells by flow cytometry, using anti-HAdV-36 hyperimmune rabbit serum. Furthermore, E1a and E4orf1 genes in long-term infected passages showed a decreasing trend. Our preliminary results reveal that renal epithelial monkey cells are permissive for the productive infection of HAdV-36. Vero cell culture long-term infection might be a promising model for addressing the fundamental aspects of the HAdV-36 biology that cannot reveal broadly-used cultures, which do not maintain long-term infection in primary or transformed cells.

Association between cycle threshold (Ct ) values and clinical and laboratory data in inpatients with COVID-19 and asymptomatic health workers.

In-house assays for the diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by quantitative reverse-transcription polymerase chain reaction (qRT-PCR), are feasible alternatives, particularly in developing countries. Cycle threshold (Ct ) values obtained by qRT-PCR were compared with clinical and laboratory data from saliva of inpatients with COVID-19 and asymptomatic health workers (AHW) were studied. Saliva specimens from 58 inpatients confirmed by qRT-PCR for SARS-CoV-2 using nasopharyngeal specimens, and 105 AHW were studied by qRT-PCR using three sets of primers for the N (N1, N2, and N3) gene of SARS-CoV-2, according to the CDC Diagnostic Panel protocol, showing a positivity of 88% for inpatients and 8% for AHW. Bivariate analysis revealed an association between Ct < 38.0 values for N2 and mechanical ventilation assistance among patients (p = .013). In addition, values of aspartate-transaminase, lactate dehydrogenase, and ferritin showed significant correlations with Ct values of N1 and N3 genes in inpatients. Therefore, our results show that Ct values correlate with some relevant clinical data for inpatients with COVID-19.

Interaction between human mucins and parasite glycoproteins: the role of lectins and glycosidases in colonization by intestinal protozoa.

Intestinal mucins are the first line of defense against microorganisms. Although knowledge about the mechanisms involved in the establishment of intestinal protozoa is limited, there is evidence that these parasites produce lectin-like molecules and glycosidases, that exert both, constitutive and secretory functions, promoting the establishment of these microorganisms. In the present review, we analyse the main interactions between mucins of the host intestine and the four main protozoan parasites in humans and their implications in intestinal colonization. There are lectin-like molecules that contain complex oligosaccharide structures and N-acetylglucosamine (GlcNAc), mannose and sialic acid as main components, which are excreted/secreted by Giardia intestinalis, and recognized by the host using mannose-binding lectins (MBL). Entamoeba histolytica and Cryptosporidium spp. express the lectin galactose/N-acetyl-D-galactosamine, which facilitates their adhesion to cells. In Cryptosporidium, the glycoproteins gp30, gp40/15 and gp900 and the glycoprotein lectin CpClec are involved in protozoan adhesion to intestinal cells, forming an adhesion-attack complex. G. intestinalis and E. histolytica can also produce glycosidases such as ß-N-acetyl-D-glucosaminidase, α-d-glucosidase, ß-d-galactosidase, ß-l-fucosidase, α-N-acetyl-d-galactosaminidase and ß-mannosidase. In Blastocystis, α-D-mannose, α-D-glucose, GlcNAc, α-D-fucose, chitin and sialic acid that have been identified on their surface. Fucosidases, hexosaminidases and polygalacturonases, which may be involved in the mucin degradation process, have also been described in the Blastocystis secretoma. Similarly, symbiotic coexistence with the intestinal microbiota promotes the survival of parasites facilitating cell invasion and nutrients obtention. Furthermore, it is necessary to identify and characterize more glycosidases, which have been only partially described by in silico analyses of the parasite genome.

Clinical study and some molecular features of Mexican patients with syndromic craniosynostosis.

BACKGROUND: Craniosynostosis is one of the major genetic disorders affecting 1 in 2,100-2,500 live newborn children. Environmental and genetic factors are involved in the manifestation of this disease. The suggested genetic causes of craniosynostosis are pathogenic variants in FGFR1, FGFR2, FGFR3, and TWIST1 genes. METHODS: In order to describe their major clinical characteristics and the presence of pathogenic variants, a sample of 36 Mexican patients with craniosynostosis diagnosed as: Crouzon (OMIM 123,500), Pfeiffer (OMIM 101,600), Apert (OMIM 101,200), Saethre-Chotzen (OMIM 101,400), and Muenke (OMIM 602,849) was analyzed. RESULTS: In addition to craniosynostosis, most of the patients presented hypertelorism, midface hypoplasia, and abnormalities in hands and feet. To detect the pathogenic variants p.Pro252Arg FGFR1 (OMIM 136,350), p.Ser252Trp, p.Pro253Arg FGFR2 (OMIM 176,943), p.Pro250Arg, FGFR3 (OMIM 134,934), and p.Gln119Pro TWIST1 (OMIM 601,622), PCR amplification and restriction enzyme digestion were performed. Four and two patients with Apert presented the pathogenic variants p.Ser252Trp and p.Pro253Arg in FGFR2, respectively (with a frequency of 11.1% and 5.5%). The p.Pro250Arg pathogenic variant of FGFR3 was found in a patient with Muenke (with a frequency of 2.8%). The above percentages were calculated with the total number of patients. CONCLUSION: The contribution of this work is discreet, since only 4 genes were analyzed and sample size is small. However, this strategy could be improved by sequencing the FGFR1, FGFR2, FGFR3, and TWIST1 genes, to determine different pathogenic variants. On the other hand, it would be important to include other genes, such as TCF12 (OMIM 600,480), MSX2 (OMIM 123,101), RAB23 (OMIM 606,144), and EFNB1 (OMIM 300,035), to determine their participation in craniosynostosis in the Mexican population.

Trypanosoma cruzi infection associated with atypical clinical manifestation during the acute phase of the Chagas disease.

BACKGROUND: Chagas disease (CD) is caused by the protozoan parasite Trypanosoma cruzi and is transmitted by triatomine insects. Clinical manifestations vary according to the phase of the disease. Cutaneous manifestations are usually observed in the acute phase (chagoma and Romaña's sign) or after reactivation of the chronic phase by immunosuppression; however, a disseminated infection in the acute phase without immunosuppression has not been reported for CD. Here, we report an unusual case of disseminated cutaneous infection during the acute phase of CD in a Mexican woman. METHODS: Evaluation of the patient included a complete clinical history, a physical exam, and an exhaustive evaluation by laboratory tests, including ELISA, Western blot and PCR. RESULTS: Skin biopsies of a 50-year-old female revealed intracellular parasites affecting the lower extremities with lymphangitic spread in both legs. The PCR tests evaluated biopsy samples obtained from the lesions and blood samples, which showed a positive diagnosis for T. cruzi. Partial sequencing of the small subunit ribosomal DNA correlated with the genetic variant DTU II; however, serological tests were negative. CONCLUSIONS: We present a case of CD with disseminated skin lesions that was detected by PCR and showed negative serological results. In Mexico, an endemic CD area, there are no records of this type of manifestation, which demonstrates the ability of the parasite to initiate and maintain infections in atypical tissues .

Association of Cystathionine ß-Synthase Gene Polymorphisms With Preeclampsia.

Preeclampsia (PE) is a pregnancy disorder that increases maternal and fetal morbidity and mortality worldwide. High plasma levels of homocysteine (Hcy) are a risk factor for several cardiovascular diseases. Cystathionine ß-synthase (CBS) plays an important role in Hcy homeostasis catalyzing the irreversible degradation of Hcy to cystathionine, protecting the endothelium from injury caused by hypoxia. Several mutations and polymorphisms may alter the expression of the CBS gene, resulting in variable levels of Hcy. The purpose of this study was to investigate the association of CBS gene polymorphisms with PE in Mexican women. A case-control study consisting of 129 pregnant women with PE (37 severe and 92 mild) and 173 women with uncomplicated pregnancies was performed. Polymorphisms, such as G797A, C785T, T833C, G919A, T959C, C1105T, and 844ins68 base pair, in the CBS gene were genotyped. The polymorphism G797A was monomorphic in cases with the presence of only G797A-G allele. Allele C785T-T and genotype C785T-C/T were associated with susceptibility in severe and mild PE. Alleles G797A-G and T959C-T were associated with susceptibility only in severe PE. Haplotype TGTWGTC was of susceptibility for severe PE and of protection for mild PE. Haplotypes CGTWGCC and CATWGTC seem to be protective for severe PE, but the latter is related to susceptibility in mild PE. The results suggest that C785T, G797A, and T959C mutations are contributing in different ways in severe and mild PE in our population and could be count as another related factor for this disease.

Is the genetic variability of Cathepsin B important in the pathogenesis of Blastocystis spp.?

The potential role of Blastocystis as a pathogen is controversial because it is found in both symptomatic and asymptomatic carriers. Since Cathepsin B has been identified as a main virulence factor that contributes to the pathogenesis of this parasite, the purpose of this study was to analyze the genetic polymorphisms of cathepsin B from Blastocystis from patients with irritable bowel syndrome and from asymptomatic carriers. DNA from fecal samples of both groups, which were previously genotyped by 18S sequencing, was used to amplify a fragment of the cathepsin B gene. Phylogenetic reconstructions were performed and some genetic population indexes were obtained. Amplicons of 27 samples (15 cases, 10 controls, and two commercial ATCC strains) were obtained and analyzed. Phylogenetic reconstructions using nucleotides or inferred amino acid sequences did not separate between cases or controls or among subtypes. Regarding the values of genetic variability, we found that the haplotype and nucleotide diversity indexes of cathepsin B from cases and controls were similar to the values of 18S from controls. By contrast, 18S from cases showed low variability, suggesting that the genetic variability of cathepsin B was not related to the symptomatology of Blastocystis carriers. However, since no polymorphisms related to cases or controls were found, it is logical to assume that the potential damage caused by Blastocystis in situ may be due to unclear mechanisms of Cathepsin B regulation and expression that should be studied in future studies.

Effect of different sources of non-fiber carbohydrate on ruminal pH and in vitro digestibility of tropical forage / Efeito de diferentes fontes de carboidratos não-fibrosos sobre o pH ruminal e digestibilidade in vitro de forragens tropicais

The present study aimed to evaluate non-fiber carbohydrates (NFC) in sugarcane-based diets on rumen pH, and forage digestibility, and to describe NFC degradation curves. The study consisted of two trials. For the first trial, three rumen cannulated steers, BW of 350 ± 15 kg (mean ± SE), were assigned in a 3×3 Latin Square (LS) design. They were fed diets containing finely-ground (0.9 mm average particle size) corn (GC), steam-rolled corn (SRC), or pelleted citrus pulp (PCP). Each period had 14 d, with the first 12 for adaptation. The 13th d was for serial measurement of rumen pH, and the14th for rumen fluid collection and in vitro incubation for DM and NDF digestibility (IVDMD and IVNDFD) of bermudagrass hay (Hay), corn (CS), and sugarcane (SS) silages. In the second trial, rumen fluid of a cannulated bull, fed corn silage and a regular concentrate, was collected for in vitro digestion of NFC for multiple time points. The incubation results were used to adjust the NFC degradation curves, and calculate lag-time, feed fractions, and degradation rate. Data from first trial was analyzed in a 3×3 LS. The model for the digestibility parameters included fixed effects of forage (Feed), diets with NFC (Diet), and their interaction (Feed × Diet), and random effect of animal and period. The model for rumen pH included fixed effect of diet, time as repeated measures, animal and period as random effects. The significance was considered at probability ≤ 5% (α = 0.05). The NFC degradation curves were adjusted using the PROC NLIN procedure from SAS, and equation parameters compared using confidence intervals. There was a Diet × Time interaction on rumen pH (P = 0.04), where SRC decreased pH compared to PCP and GC diets at the time 6 h, only. There was no Feed × Diet interaction effect (P > 0.05) for any digestibility parameter. There was a Feed effect on both IVDMD and IVNDFD, either after 30 or 48 h incubation (P < 0.01). The CS had the greatest IVDMD, followed by SS and Hay, after 30 and 48 h of incubation. The CS had the greatest IVNDFD after 30 h, compared to SS and Hay. However, for IVNDFD after 48 h, CS presented the greatest mean, followed by SS and Hay. The rumen fluid from animals fed SRC decreased both IVDMD and IVNDFD (P < 0.05) of all roughages after 48 h. Results from the second trial showed that the PCP had lower Lag Time, B fraction and greater kd compared to both corn sources, and SRC had greater kd than GC. In conclusion, the SRC diet decreased rumen pH 6 h after feeding and, consequently, decreased fiber digestibility of the tropical forage sources evaluated. Although the PCP had lower lag time, and faster rate of degradation of B fraction, it did not negatively affect rumen pH or fiber digestibility of forage.(AU)

O presente estudo teve como objetivo avaliar os carboidratos não-fibrosos (CNF) em dietas à base de cana-de-açúcar sobre o pH ruminal e digestibilidade da forragem, e descrever as curvas de degradação dos CNF. O estudo foi composto de dois ensaios. No primeiro, três novilhos canulados no rúmen, com peso vivo de 350 ± 15 kg (Média ± DP), foram alocados em um quadrado latino (QL) 3×3, e alimentados com dietas contendo: milho moído (MM, tamanho de partículas 0,9 mm), laminado a vapor (MLV) ou polpa cítrica peletizada (PCP). Cada período tinha 14 d, sendo os primeiros 12 para adaptação e o 13º para a medição seriada do pH e o 14º para a coleta de líquido ruminal e incubação in vitro para digestibilidade da MS e FDN (DIVMS e DIVFDN) de feno de bermudagrass (Feno) e silagens de milho (SM) e cana (SC). No segundo ensaio, coletou-se fluido ruminal de um touro canulado, alimentado com silagem de milho e concentrado padrão, para digestão in vitro dos CNF em vários tempos. Esses resultados foram utilizados para ajustar as curvas de degradação dos CNF e calcular o tempo de colonização, frações alimentares e taxa de degradação. Os resultados do primeiro ensaio foram analisados em um QL 3×3. O modelo dos parâmetros de digestibilidade incluiu efeito fixo de forragem (Alimento), dieta com CNF (Dieta) e interação (Alimento × Dieta), e efeito aleatório de animal e período. O modelo para pH incluiu efeito fixo de Dieta, Tempo como medida repetida, animal e período como aleatórios. Foi considerada a probabilidade significativa de ≤ 5% (α = 0,05). As curvas de degradação dos CNF foram ajustadas pelo PROC NLIN do SAS, e parâmetros de equação comparados por intervalo de confiança. Houve interação Dieta × Tempo no pH ruminal (P = 0,04), onde o MLV diminuiu o pH comparado com PCP e MM apenas no tempo 6 h. Não houve interação Alimento × Dieta (P > 0,05) para nenhum parâmetro de digestibilidade. Houve efeito de Alimento sobre a DIVMS e DIVFDN, após 30 e 48 h de incubação (P < 0,01). A SM teve a maior DIVMS, seguido por SC e Feno, após 30 e 48 h de incubação. A SM teve a maior DIVFDN após 30 h, comparado com SC e Feno. No entanto, para DIVFDN após 48 h, a SM teve maior média, seguida da SC e Feno. O fluido ruminal de animais alimentados com MLV diminuiu a DIVMS e DIVFDN (P < 0.05) de todas as forragens, após 48 h. Resultados do segundo ensaio mostram que PCP diminuiu o tempo de colonização, fração B e aumentou a kd comparado com os dois milhos, e MLV apresentou maior kd que o MM. Em conclusão, a dieta com MLV diminuiu o pH ruminal no tempo 6 h e, consequentemente, diminuiu a DIVFDN das forragens avaliadas. Embora PCP tenha apresentado menor tempo de colonização e maior taxa de degradação da fração B, não afetou negativamente o pH do rúmen nem a digestibilidade da fibra das forragens. (au)