Polimorfismo de los genes que codifican para las proteínas VP4, VP7 y NSP4 de rotavirus y su asociación con la severidad de la diarrea

De los virus que provocan gastroenteritis, rotavirus es el principal agente causal de diarreas severas provocando la muerte de cerca 440,000 niños por año a nivel mundial. Algunos estudios señalan a las proteínas VP4, VP7 y NSP4 como factores de patogenicidad; VP4 y VP7 se involucran en procesos de ingreso del virus a la célula y en la producción de anticuerpos de neutralización; NSP4 es la única proteína descrita como enterotoxina, la cual, mediante un incremento en Ca2+ desencadena los procesos diarreicos. Hasta el momento, no se conocen los mecanismos por los cuales rotavirus puede causar una gastroenteritis severa. Por lo que el objetivo del presente trabajo es determinar si existe una correlación entre el polimorfismo de los genes que codifican para VP4, VP7 y NSP4 con los casos severos de diarrea. Se analizaron 123 muestras de heces de niños menores de 5 años colectadas de Octubre de 2004 a Marzo de 2005. La presencia del RNA característico de rotavirus se determinó a través de electroforesis en un sistema continuo. Posteriormente el RNA se sometió a RT-PCR para determinar los genotipos VP4, VP7 y NSP4. Los productos de PCR fueron desnaturalizados y sometidos a un sistema de SSCP para detectar el polimorfismo intragenotípico de las cepas de estudio y someter a una población representativa a secuencia de nucleótidos. Los resultados muestran que 64 (52%) de las cepas presentaron el modelo de migración característico de rotavirus. Con respecto a VP7 y VP4 el genotipo que predominó fue G1P[8] (34%), para NSP4 el genotipo Wa (65%) fue el que más se presentó. Por otro lado, las variaciones intragenotípicas detectadas en los genes que codifican para VP4 y VP7 mostraron tres modelos de migración y no se detectó una asociación con la severidad de la gastroenteritis. Por su parte, para NSP4, se detectaron 5 modelos de variación intragenotípica, mismos que se correlacionaron con el grado de deshidratación (P<0.05). Los análisis de secuencia de los modelo indicó variaciones en la regiones de aa 115- 140 para los modelos 1,2,3 y 5; pero no para el modelo 4, mismo que se relacionó con los casos más severos. Estos resultados muestran que VP7 y VP4 no están involucrados en la severidad de la infección y que NSP4 además de ser un factor de infecciones sintomáticas, como lo han reportado algunos investigadores, también puede ser un factor de la severidad de la gastroenteritis, indicando la importancia de estudiar este gen y la proteína que codifica para tener un mayor conocimiento de la patogénesis de la gastroenteritis por rotavirus. Abstract Of the viruses that cause gastroenteritis, rotavirus is the main causal agent of severe diarrhea; it causes the fence death 440,000 children per year at world-wide level. Some researchers point to proteins VP4, VP7 and NSP4 like pathogenicity factors; VP4 and VP7 are related with the penetration viral into epithelial cells and the synthesis of neutralization antibodies; NSP4 is the first described viral enterotoxin, which, by means of an increase in Ca2+ it triggers the diarrheic processes. At the moment, are not known the mechanisms by which rotavirus can cause a severe gastroenteritis. The goal of the present work is determine if exist a correlation between the polymorphism of the genes codifies for VP4, VP7 and NSP4 with the severe cases of diarrhea. On this study were analyzed 123 samples of diarrheic faeces of children less than five years old. The presence of the RNA characteristic of rotavirus was determined by electroforesis in a continuous system. The VP4, VP7 and NSP4 genotypes were determined through RT-PCR. To detect the intragenotípic polymorphism, the PCR products were analyzed by SSCP. The results show that 64 (52%) of the samples presented the model of migration characteristic of rotavirus. The genotypes G1P[8] (34%) and Wa (65%) were predominant in VP7-VP4 and NSP4, respectively. On the other hand, the detected intragenotipic variations in the genes that codify to VP4 and VP7 was not associated to the severity of the gastroenteritis; it was not the case for NSP4, where the five different models from intragenotipic variation were detected, same that was correlated with the dehydration degree (P<0.05). The analysis of sequence of the models showed variations in the regions of aa 115-140 for the models 1, 2,3 and 5; but it does not to model 4, same that was related to the most severe cases. These results show that probably VP7 and VP4 are determining for the infection and the viral neutralization, but not thus for the severity of the gastroenteritis. On the other hand, NSP4 can influence in the presence of asymptomatic infections, it reported by some investigators, if not also in the severity of the gastroenteritis, indicating that NSP4 can be considered like an important factor of viral pathogenicity in natural infections

Mutation Distribution in the NSP4 Protein in Rotaviruses Isolated from Mexican Children with Moderate to Severe Gastroenteritis

The NSP4 protein is a multifunctional protein that plays a role in the morphogenesis and pathogenesis of the rotavirus. Although NSP4 is considered an enterotoxin, the relationship between gastroenteritis severity and amino acid variations in NSP4 of the human rotavirus remains unclear. In this study, we analyzed the sequence diversity of NSP4 and the severity of gastroenteritis of children with moderate to severe gastroenteritis. The rotavirus-infected children were hospitalized before the rotavirus vaccine program in Mexico. All children had diarrhea within 14 days, 44 (88%) were vomiting and 35 (70%) had fevers. The severity analysis showed that 13 (26%) cases had mild gastroenteritis, 23 (46%) moderate gastroenteritis and 14 (28%) severe. NSP4 phylogenetic analysis showed three clusters within the genotype E1. Sequence analysis revealed similar mutations inside each cluster, and an uncommon variation in residue 144 was found in five of the Mexican NSP4 sequences. Most of the amino acid variations were located in the VP4 and VP6 binding site domains, with no relationship to different grades of gastroenteritis. This finding indicates that severe gastroenteritis caused by the rotavirus appears to be related to diverse viral or cellular factors instead of NSP4 activity as a unique pathogenic factor

Modulation of rotavirus severe gastroenteritis by the combination of probiotics and prebiotics

Abstract Annual mortality rates due to infectious diarrhea are about 2.2 million; children are the most vulnerable age group to severe gastroenteritis, representing group A rotaviruses as the main cause of disease. One of the main factors of rotavirus pathogenesis is the NSP4 protein, which has been characterized as a viral toxin involved in triggering several cellular responses leading to diarrhea. Furthermore, the rotavirus protein NSP1 has been associated with interferon production inhibition by inducing the degradation of interferon regulatory factors IRF3, IRF5, and IRF7. On the other hand, probiotics such as Bifidobacterium and Lactobacillus species in combination with prebiotics such as inulin, HMO, scGOS, lcFOS have been associated with improved generalized antiviral response and anti-rotavirus effect by the reduction of rotavirus infectivity and viral shedding, decreased expression of NSP4 and increased levels of specific anti-rotavirus IgAs. Moreover, these probiotics and prebiotics have been related to shorter duration and severity of rotavirus diarrhea, to the prevention of infection and reduced incidence of reinfections. In this review we will discuss in detail about the rotavirus pathogenesis and immunity, and how probiotics such as Lactobacillus and Bifidobacterium species in combination with prebiotics have been associated with the prevention or modulation of rotavirus severe gastroenteritis. Keywords Rotavirus · Pathogenesis · Probiotics · Prebiotic

Rendimiento y valor nutricional de brásicas forrajeras en comparación con forrajes tradicionales

The high nutritional value of brassicas can increase productivity in traditional forage production systems. The objective of the study was to compare the nutritional value and yield of dry matter (DM) and nutrients between forage brassicas and traditional autumn-winter species. The forage brassicas were Winfred, Hunter and Graza radish and the traditional forages were oats, triticale, barley, wheat and berseem clover. The study was conducted in Matamoros, Coahuila, Mexico in the 2018-2019 cycle, under a randomized complete block experimental design with four repetitions. The regrowth capacity, the nutritional composition of the forage and the yields of DM and nutrients were determined. All species showed regrowth capacity, with three cuts in berseem clover in 154 d, and with two cuts in brassicas (150-154 d) and cereals (133-144 d). The brassicas had nutritional composition similar to that of berseem clover and better than that of cereals, mainly due to their higher content of net energy of lactation (NEL; 6.57 to 7.32 MJ kg-1 DM). The DM yields of the brassicas were similar to those observed in traditional forages; however, due to their high nutritional composition, the brassicas were equal to or superior in production of crude protein (CP) (1,608 to 2,986 kg ha-1) and NEL (62,819 to 84,044 MJ ha-1) to traditional forages. In general, forage brassicas can increase nutrient yield with respect to cereals and berseem clover, especially in the production of NEL (27.5 to 47.3 %).El alto valor nutritivo de las brásicas puede incrementar la productividad en los sistemas de producción de forrajes tradicionales. El objetivo del estudio fue comparar el valor nutricional y el rendimiento de materia seca (MS) y nutrientes entre brásicas forrajeras y especies tradicionales de otoño-invierno. Las brásicas forrajeras fueron Winfred, Hunter y rábano Graza y los forrajes tradicionales fueron avena, triticale, cebada, trigo y el trébol Alejandrino. El estudio se realizó en Matamoros, Coahuila, México en el ciclo 2018-2019, bajo un diseño experimental de bloques completos al azar con cuatro repeticiones. Se determinó la capacidad de rebrote, la composición nutricional del forraje y los rendimientos de MS y nutrientes. Todas las especies presentaron capacidad de rebrote, con tres cortes en trébol Alejandrino en 154 días, y con dos cortes en brásicas (150-154 días) y cereales (133-144 días). Las brásicas presentaron similar composición nutricional al trébol Alejandrino y mejor al de los cereales, principalmente por su mayor contenido de energía neta para lactancia (ENL; 6.57 a 7.32 MJ kg-1 MS). Los rendimientos de MS de las brásicas fueron similares a los observados en los forrajes tradicionales; sin embargo, por su alta composición nutricional las brásicas fueron iguales o superiores en producción de proteína cruda (PC) (1,608 a 2,986 kg ha-1) y ENL (62,819 a 84,044 MJ ha-1) a los forrajes tradicionales. En general, las brásicas forrajeras pueden incrementar el rendimiento de nutrientes respecto a los cereales y al trébol Alejandrino, especialmente en la producción de ENL (27.5 a 47.3 %)

Multimodal treatment of myophasal chronic pelvic pain in women: institutional experience

Background: Chronic pelvic pain of myofascial origin is an underdiagnosed condition and with partial responses to the different treatment modalities. The objective of this study was to report the results of the multimodal management of Myofascial Chronic Pelvic Pain.Methods: Retrospective cohort of 33 patients with myofascial chronic pelvic pain, during the period 01 January 2016 to August 31, 2018, who were treated based on trigger point infiltration and analgesic vaginal electrostimulation. The analogous visual pain scale (VAS) was used at the beginning, sixth, twelfth and six months after the last electrostimulation session, in order to measure the effect of the treatment.Results: During the study period, 32 patients with chronic myofascial pelvic pain were found who received the multimodal scheme that is offered in our clinic. The average score of the EVA before starting the treatment was 8.5 points, at the sixth session of the treatment the average EVA was 3.6 points, at the twelfth session of 1.3 points and the average score of the EVA at six months after the last session of the multimodal treatment was 2.9 points. A significant difference was demonstrated in the average EVA score in the first with respect to the sixth, twelfth and six months (p = <0.000). There were no complications or adverse effects secondary to the multimodal treatment.Conclusions: Multimodal treatment was associated with progressive improvement in 80% of patients with miofascial chronic pelvic pain

Optimization of total anthocyanin content and antioxidant activity of a Hibiscus sabdariffa infusion using response surface methodology

Hibiscus sabdariffa L. calyces are underutilized sources of health-promoting anthocyanins. Infusions are the most common way to consume them, but because anthocyanins are thermosensitive, prolonged extraction times at high temperatures may reduce their bioactivities, suggesting the need to identify optimal preparation conditions. Response surface methodology was used to establish calyces-to-water ratio (X1: 1–20 g/100 mL), temperature (X2: 70–100 °C), and time (X3: 1–30 min) that would produce an infusion with optimized total anthocyanin content (TAC) and antioxidant activity. Under optimum conditions (X1=10 g/100 mL, X2=88.7 °C, and X3=15.5 min) TAC was 132.7±7.8 mg cyanidin-3-glucoside equivalents (C3G)/100 mL, and antioxidant activity was 800.6±69.9 (DPPH assay), and 1792.0±153.5 (ABTS assay) μmol Trolox equivalents (TE)/100 mL. Predicted and experimental results were statistically similar. Identifying ideal processing conditions can promote consumption of an H. sabdariffa-based functional beverage with high anthocyanin content and antioxidant activity that exert health-promoting bioactivities on the consumer

Bioactive Products From Plant-Endophytic Gram-Positive Bacteria

Endophytes constitute plant-colonizing microorganisms in a mutualistic symbiosis relationship. They are found in most ecosystems reducing plant crops’ biotic and abiotic stressors by stimulating immune responses, excluding plant pathogens by niche competition, and participating in antioxidant activities and phenylpropanoid metabolism, whose activation produces plant defense, structural support, and survival molecules. In fact, metabolomic studies have demonstrated that endophyte genes associated to specific metabolites are involved in plant growth promotion (PGP) by stimulating plant hormones production such as auxins and gibberellins or as plant protective agents against microbial pathogens, cancer, and insect pests, but eco-friendly and eco-safe. A number of metabolites of Gram-positive endophytes isolated from agriculture, forest, mangrove, and medicinal plants, mainly related to the Firmicutes phyla, possess distinctive biocontrol and plant growth-promoting activities. In general, Actinobacteria and Bacillus endophytes produce aromatic compounds, lipopeptides, plant hormones, polysaccharides, and several enzymes linked to phenylpropanoid metabolism, thus representing high potential for PGP and crop management strategies. Furthermore, Actinobacteria have been shown to produce metabolites with antimicrobial and antitumor activities, useful in agriculture, medicine, and veterinary areas. The great endophytes diversity, their metabolites production, and their adaptation to stress conditions make them a suitable and unlimited source of novel metabolites, whose application could reduce agrochemicals usage in food and drugs production

Evaluation of chromosome organization and microtubule arrangement in goat (capra aegragrus) oocytes after vitrification, in vitro maturation and fertilization, and early embryo development

Objective: Evaluate the use of Ethylene Glycol (EG), Dimethyl Sulfoxide (DMSO), Sucrose and Fetal Bovine Serum (FBS) as cryoprotectants and their effect on the organization of chromosomes and the arrangement of microtubules, during the vitrification process in goat oocytes matured in vitro and in the development of preimplantation embryos produced in vitro. Design/methodology/approach: In vitro matured oocytes were divided into 3 groups (control group, cryoprotectant exposed group, vitrified group). A mixture of 15% EG, 15% DMSO, 0.4 M sucrose and 20% FBS was used for the vitrification using the Cryotop device. In vitro matured oocytes were warmed and afterwards each group was divided into two more groups. Both groups were subjected to immunofluorescence, the first group to observe the damage produced to the chromosomes and microtubules and the second group to observe the effect on the in vitro embryo development. Results: The combined use of 15% EG, 15% DMSO, 0.4 M Sucrose and 20% FBS during vitrification did not prevent cryoinjuries in goat oocytes and in vitro produced embryos, since embryo development was disrupted before the blastocyst stage by stopping cleavage at the morula stage. This disruption was associated with chromosome decondensation and the absence of a microtubule network, thereby hindering chromosomal segregation. Limitations on study/implications: The effect of conventional cryoprotectants on chromosomes and microtubules arrangement on vitrified goat oocytes and in vitro embryo production. Findings/conclusions: The combined use of 15% EG, 15% DMSO, 0.4 M sucrose and 20% FBS as vitrification cryoprotectants did not prevent cryoinjuries in caprine oocytes and did not improve caprine embryo development in vitro

Reflexiones y aportaciones metodológicas

los aportes reunidos en esta obra, provienen de experiencias previas de trabajo profesional y de investigación, lo cual es resultado de distintos enfoques y vivencias de los grupos de investigación aquí involucrados, los que han interactuado en distintos eventos académicos y específicamente, en seminarios que propiciaron la producción que se reúne en esta publicación colectiva. Finalmente, esta experiencia acumulada es una de las mejores recomendaciones para la lectura por demás interesante y propositivo.El presente libro, identifica los aportes metodológicos del trabajo cotidiano en diferentes capítulos para integrarlos a los propios de las disciplinas del diseño. Asimismo, uno de los capítulos aborda la necesidad de estudiar un contexto de referencia que el destinatario pueda interpretar, entre un canal físico y una conexión psicológica a modo de estrategia interpretativa individual y colectiva. Este discurso, sostienen los coautores, se enmarca en la hermenéutica y dentro de lo colectivo, ya que este proceso se estudia desde las aportaciones del interaccionismo simbólico. Se abandona la construcción de las utopías, a la vez que busca soluciones diseñísticas, actuales y novedosas. Es así que el sentido del mensaje del objeto de diseño como texto conlleva un discurso que no “es realizado por el emisor, sino por el receptor y es la enunciación, narración y reconfiguración de los textos, en donde el discurso conforma una determinada idea de la realidad”