Diseño y construcción de un péptido con potencial inmunogénico a partir del dominio de unión a supresor de hairless de la proteína hairless de Drosophila melanogaster

Signal transduction pathways regulate the functions of different types of cells that constitute a multicellular organism, with the final goal of to regulate of target genes. The Notch signaling pathway is mainly involved in early and late embryo development, and recently has been described its participation in the adult organism proper function. Hairless, a protein member of the Notch signaling pathway modulates negatively this signal transduction pathway. Despite efforts to detect Hairless in higher animals, it has not been able to identify a homolog protein. The identification of a Hairless homolog protein is essential for the understanding of the regulatory processes during embryo development and other pathological processes where Notch pathway is involved. The Hairless counterparts described in insects have at least three highly conserved domains, where the Suppressor of Hairless binding domain is the most conserved. Using analysis in silico and molecular biology techniques was identified and delimited a peptide with immunogenic potential from the Suppressor of Hairless binding domain from Drosophila melanogaster Hairless protein. This peptide was used as antigen to generate polyclonal antibodies in hens. Sera were tested against total protein extract from fruit fly embryos.Las funciones de los diferentes tipos de células que constituyen un organismo multicelular, son reguladas mediante vías de transducción de señales, cuyo objetivo final es la regulación de genes blanco. La vía de señalización Notch participa principalmente en el desarrollo temprano y tardío del embrión, y en menor grado en el funcionamiento correcto del individuo. Hairless es una proteína involucrada en la vía de señalización Notch y su papel es modular negativamente la vía. A pesar de los esfuerzos realizados para detectar a hairless en animales superiores, no se ha logrado identificar un homólogo de la misma. Su identificación es fundamental para entender mejor los procesos de regulación de la vía, del desarrollo embrionario y de otros procesos patológicos que involucran a Notch. Los homólogos de Hairless en insectos poseen al menos tres dominios altamente conservados, de los cuales el dominio de unión a Supresor de hairless, una proteína propia de la vía, es el más conservado. Mediante análisis in silico y técnicas de biología molecular se identificó y delimitó un péptido con potencial inmunogénico del dominio de unión a Supresor de hairless de la proteína hairless de Drosophila melanogaster. Este péptido se usó como antígeno en gallinas, para generar anticuerpos policlonales. Los sueros se probaron contra extracto total de proteína de embriones de la mosca de la fruta

Caracterización molecular de aislamientos de Staphylococcus spp. asociados a mastitis bovina en Tarímbaro, Michoacán

Mastitis is a disease of dairy cattle worldwide that causes decreased milk quality/yield, resulting in major economic losses. Mastitis is caused by a wide variety of organisms, including Staphylococcus aureus as a major pathogen. The purpose of this research was to characterize 20 Staphylococcus spp. isolates from the mammary gland of mastitic, lactating, backyard cattle, using both the Random Amplified Polymorphic DNA (RAPD) analysis, and sensitivity tests to 13 antibiotics and 1 detergent. Sensitivity tests to 13 antibiotics showed that all isolates were resistant to penicillin but sensitive to cephalothin, erythromycin, and trimethoprim. All isolates were susceptible to a 90% similarity in the RAPD band pattern, while isolates SA35 & SA36 were identical to each other. Nevertheless, all other isolates showed different amplification profiles. Isolates SA7 & SA8, SA30 & SA32, SA35 & SA36 had similar antibiotic resistance/RAPD patterns, suggesting re-isolation. Staphylococcus isolates collected in Tarímbaro, Michoacán showed several different amplification patterns.La mastitis es una enfermedad que afecta al ganado bovino lechero a nivel mundial, provocando una disminución en la calidad y cantidad de la leche producida, lo cual genera pérdidas económicas considerables. Esta enfermedad es causada por una variedad de microorganismos, entre los cuales Staphyloccocus aureus es uno de los más importantes. El objetivo del trabajo fue caracterizar 20 aislamientos de Staphylococcus spp. obtenidos de la glándula mamaria de vacas lactantes con mastitis en un sistema de producción de traspatio, mediante el uso de polimorfismos derivados de la amplificación al azar de ADN (RAPD's) y su sensibilidad hacia trece antibióticos y 1 detergente. Las pruebas de sensibilidad a los 13 antibióticos demostraron que todos los aislamientos fueron resistentes a penicilina y sensibles a cefalotina, eritromicina y trimetoprim. Todos los aislamientos fueron susceptibles a una concentración menor de 6 µg/ml de bromuro de cetil trimetil amonio (CTAB). La caracterización molecular permitió determinar que los aislamientos SA7 y SA8, SA30 y SA32, presentaron una similitud superior al 90 % en el patrón de bandas obtenida por RAPD, mientras que los aislamientos SA35 y SA36 fueron idénticos, sin embargo, el resto de los aislamientos presentaron perfiles de amplificación distintos. Los aislamientos SA7 y SA8, SA30 y SA32, SA35 y SA36, tuvieron patrones de resistencia a antibióticos y de RAPD similares, por lo que probablemente se trató de reaislamientos. Los aislamientos de Staphylococcus de Tarí­mbaro, Michoacán presentaron una diversidad de perfiles de amplificación

Behavior of Eosinophil Counts in Recovered and Deceased COVID-19 Patients over the Course of the Disease

Knowledge about the immune responses against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, particularly regarding the function of eosinophils, has been steadily emerging recently. There exists controversy regarding the implications of eosinophils in the coronavirus disease 2019 (COVID-19)’s pathology. We report a retrospective cohort study including the comparison of leukocyte counts in COVID-19 patients, considering the outcomes of recovery (n = 59) and death (n = 60). Among the different types of leukocytes, the eosinophil counts were those that showed the greatest difference between recovered and deceased patients. Eosinopenia (eosinophil count < 0.01 × 109/L) was more frequently observed in deceased than recovered patients (p = 0.0012). The eosinophil counts more rapidly increased and showed a greater proportion over the course of the disease in the recovered than deceased patients. Furthermore, the estimated survival rate was greater in patients without eosinopenia than in patients with eosinopenia (p = 0.0070) during hospitalization. Importantly, recovered but not deceased patients showed high negative correlations of the eosinophils with the neutrophil-to-lymphocyte ratio (NLR) and neutrophil counts at Day 9 of the onset of clinical symptoms (p ≤ 0.0220). Our analysis suggests that eosinopenia may be associated with unfavorable disease outcomes and that the eosinophils have a beneficial function in COVID-19 patients, probably contributing by controlling the exacerbated inflammation induced by neutrophils