RESUMEN
Therapy with biopharmaceuticals, mainly recombinant antibodies, offers patients higher life expectancy and better life quality than pharmacologic therapy. Countries with the highest scientific development are investing in this kind of therapy, and this is why the optimization of the production of these recombinant proteins would lead to their higher production and lower costs of the final product. Modifications in the use of promoters, the use of recombination regions, and the change in the order of the chains, are some of the genetic engineering changes that can increase the production of recombinant antibodies. In this work, three different promoters were tested: Prom A, hCMV, and EF1-a, for two different antibodies, one anti-TNFa and one anti-CD20+. Changes were made in the order of the chains H-L or L-H and one or two UCOE (ubiquitous chromatin opening element) sequences were also used to identify the combinations that provide the best transient and stable expression for the antibodies in the CHO-s cells. In our results, we observed that the use of the two UCOE regions, with L-H order is almost three times better for the expression of the two different antibodies, while the strength of the promoter is conditioned by the sequence of each expressed protein.
Asunto(s)
Anticuerpos Monoclonales , Antígenos CD20 , Expresión Génica , Regiones Promotoras Genéticas/genética , Factor de Necrosis Tumoral alfa , Animales , Anticuerpos Monoclonales/biosíntesis , Anticuerpos Monoclonales/genética , Células CHO , Cricetinae , Cricetulus , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genéticaRESUMEN
INTRODUCTION: Coronavirus disease 19 (COVID-19) has been a global public health emergency, with 209.89 million cases of infection with SARS-CoV-2 recorded, resulting in 4,401,675 deaths. After recuperation, it is probable that COVID-19 patients have sequelae of the disease. This study aimed to evaluate the respiratory anatomical-functional sequelae in Mexican patients who recovered from COVID-19. METHODOLOGY: This study included twenty-four patients who recovered from COVID-19 and eight non-infected patients (controls). Participants were screened for SARS-CoV-2 and the presence of IgM/IgG antibodies. Pulmonary function and lung anatomical abnormalities were evaluated by spirometry and computerized tomography. RESULTS: A total of 45.8% of the patients had pulmonary function with obstructive patterns: 70.8% of recovered cases had COVID-19 Reporting and Data System (CO-RADS) 1, 20.8% CO-RADS 3 and 16.7% CO-RADS 4. A total of 35.3% of patients with CO-RADS 1 also showed bilateral nodal growth; 70.8% of patients tested positive for IgG and 8.4% for IgG/IgM, and 20.8% tested negative for both antibodies. CONCLUSIONS: There were respiratory anatomical and functional sequelae in Mexican patients who recovered from COVID-19, with a high occurrence of pulmonary obstructive patterns in the study population. These observations indicate the importance of the routine evaluation of sequelae in Mexican patients who recovered from COVID-19 and the need for strict follow-up to improve the quality of life of these patients.
Asunto(s)
COVID-19 , Anticuerpos Antivirales , Humanos , Inmunoglobulina M , Pulmón , Calidad de Vida , SARS-CoV-2RESUMEN
INTRODUCTION: Introduction: worldwide, hospital malnutrition constitutes an important issue of morbidity and mortality. Although the prevalence of malnutrition has been calculated as between 7% and 27% in hospitalized patients, its real prevalence remains unknown or underestimated because of the different criteria for its identification and diagnosis. The aim of this study was to determine the prevalence of nutritional risk in a cohort of hospitalized patients and to identify the significance of the predictors associated with nutritional risk. Methods: the evaluation of the presence of nutritional risk was carried out in 247 individuals hospitalized at the second-level care institution Instituto de Seguridad y Servicios Sociales para los Trabajadores del Estado (ISSSTE) Zacatecas, Hospital General Nº 26, in Mexico. Nutritional screening was evaluated during the first 24 hours of stay with the NRS 2002. The weighing of associated variables with nutritional risk was calculated statistically using the software Sigma Plot v11. Results: forty-two percent of patients were at risk of malnutrition. Significant associations between nutritional risk and a reduction in food ingestion (during the last week), the illness severity of the patient, as well as age and sex (p < 0.05), were observed. A reduction in food ingestion during the previous week increased the likelihood of having nutritional risk 6.67 times more (95% CI: 3.4-13.2; p < 0.001) in the studied population. Conclusion: the risk of malnutrition in hospitalized patients at ISSSTE-Zacatecas, Hospital General Nº 26 is frequent (42%). Therefore, early detection of nutritional risk is important to offer for proper nutritional intervention with the objective of decreasing the associated morbidity and mortality.
INTRODUCCIÓN: Introducción: la desnutrición hospitalaria constituye un problema de morbimortalidad en todo el mundo. Aunque la prevalencia de malnutrición se ha calculado entre el 7% y el 27% en pacientes hospitalizados, su prevalencia real sigue siendo desconocida o subestimada debido a los diferentes criterios para su identificación y diagnóstico. El objetivo de este estudio fue determinar la prevalencia del riesgo nutricional mediante la herramienta Nutritional Risk Screening 2002 (NRS-2002) en pacientes hospitalizados y ponderar los factores predictivos asociados con el riesgo nutricional. Métodos: la evaluación de la presencia de riesgo nutricional se realizó en 247 hospitalizados en el hospital general de segundo nivel Instituto de Seguridad y Servicios Sociales para los Trabajadores del Estado (ISSSTE) Zacatecas, Hospital General Nº 26, en México. La evaluación nutricional se realizó durante las primeras 24 horas de estadía mediante la herramienta NRS 2002. El análisis de datos se llevó a cabo mediante el software Sigma Plot v11. Resultados: el 42% de los pacientes presentaron riesgo de desnutrición. Después de la corrección por covariables, se encontraron asociaciones significativas entre el riesgo nutricional y una reducción de la ingesta de alimentos (durante la última semana), la gravedad de la enfermedad del paciente, la edad y el sexo (p < 0,05). Entre la población estudiada, la reducción de la ingesta durante la última semana aumentó 6,67 veces la probabilidad de presentar riesgo nutricional (IC 95%: 3,4-13,2; p < 0,001). Conclusión: el riesgo de desnutrición en pacientes hospitalizados en el Hospital General Nº 26 Zacatecas-ISSSTE es frecuente (42%), por lo que es importante realizar una detección temprana para ofrecer una intervención nutricional adecuada y, con ello, disminuir la morbimortalidad asociada.
Asunto(s)
Desnutrición/diagnóstico , Evaluación Nutricional , Medición de Riesgo/métodos , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Índice de Masa Corporal , Estudios de Cohortes , Ingestión de Alimentos , Femenino , Hospitalización , Humanos , Masculino , Desnutrición/epidemiología , Tamizaje Masivo , México/epidemiología , Persona de Mediana Edad , Prevalencia , Factores SexualesRESUMEN
Culex quinquefasciatus Say is a vector of many pathogens of humans, and both domestic and wild animals. Personal protection, reduction of larval habitats, and chemical control are the best ways to reduce mosquito bites and, therefore, the transmission of mosquito-borne pathogens. Currently, to reduce the risk of transmission, the pyrethroids, and other insecticide groups have been extensively used to control both larvae and adult mosquitoes. In this context, amino acids and acylcarnitines have never been associated with insecticide exposure and or insecticide resistance. It has been suggested that changes in acylcarnitines and amino acids profiles could be a powerful diagnostic tool for metabolic alterations. Monitoring these changes could help to better understand the mechanisms involved in insecticide resistance, complementing the strategies for managing this phenomenon in the integrated resistance management. The purpose of the study was to determine the amino acids and acylcarnitines profiles in larvae of Cx. quinquefasciatus after the exposure to different insecticides. Bioassays were performed on Cx. quinquefasciatus larvae exposed to the diagnostic doses (DD) of the insecticides chlorpyrifos (0.001 µg/mL), temephos (0.002 µg/mL) and permethrin (0.01 µg/mL). In each sample, we analyzed the profile of 12 amino acids and 31 acylcarnitines by LC-MS/MS. A t-test was used to determine statistically significant differences between groups and corrections of q-values. Results indicates three changes, the amino acids arginine (ARG), free carnitine (C0) and acetyl-carnitine (C2) that could be involved in energy production and insecticide detoxification. We confirmed that concentrations of amino acids and acylcarnitines in Cx. quinquefasciatus vary with respect to different insecticides. The information generated contributes to understand the possible mechanisms and metabolic changes occurring during insecticide exposure.
Asunto(s)
Aminoácidos/metabolismo , Carnitina/análogos & derivados , Culex/metabolismo , Insecticidas/farmacología , Larva/metabolismo , Metaboloma , Animales , Carnitina/metabolismo , Culex/clasificación , Culex/efectos de los fármacos , Larva/efectos de los fármacos , Espectrometría de Masas en TándemRESUMEN
RESUMEN: Los instrumentos rotatorios utilizados en el tratamiento de conductos dentales necesitan cumplir con ciertas características físicas que le confieran un uso correcto, adecuado y seguro para su uso clínico. En el mercado comercial existen una serie de instrumentos importados que dejan en duda la autenticidad de estos debido a sus bajos costos. En este estudio evaluamos física, química y morfológicamente dos kits de instrumentos rotatorios Protaper universal (Dentsply) mediante análisis de MEB, EDX, microdureza y EDS. Uno de los kits (grupo 1) se adquirió directamente de la casa comercial y otro kit importado que a simple vista no parecía cumplir con los estándares de calidad de un correcto empaquetado (grupo 2). En el análisis de MEB se observaron características morfológicas muy diferentes entre ambos kits, el grupo 2 presentó diversas irregularidades en la superficie de las limas, sin embargo, en el EDS no se encontró diferencia alguna. En cuanto al análisis de microdureza se observó una estadística estadísticamente significativa y en el EDS se observaron mayores picos de intensidad en cuanto a la aleación de Ni-Ti en el grupo 2. Estos resultados sugieren que existen instrumentos rotatorios importados que a pesar de su bajo costo pueden presentar ciertas características muy similares a los kits auténticos, sin embargo, física y químicamente pueden resultar en un riesgo para su uso clínico debido a la diferencia entre estos.
ABSTRACT: Rotatory files are instruments used in the treatment of dental canals roots. These instruments need to comply with certain physical characteristics for a correct, adequate and safe use for clinical use. In the commercial market there are a series of imported instruments that cast doubt on their authenticity due to their low costs. In this study we physically, chemically and morphologically evaluated two Protaper universal rotary instrument kits (Dentsply) using SEM, EDX, microhardness and EDS analysis. One of the kits (group 1) was purchased directly from the commercial house and another imported kit that a simple view did not seem to meet the quality standards of a correct packaging (group 2). In the SEM analysis, very different morphological characteristics were observed between both kits, group 2 presented various irregularities on the surface of the files, however, no difference was found in the EDS. Regarding the microhardness analysis, a statistically significant statistic was observed and in the EDS, higher intensity peaks were observed in terms of the Ni-Ti alloy in group 2. These results suggest that there are imported rotary instruments that despite their low cost can present characteristics very similar to authentic kits, however, physically and chemically they can result in a risk for their clinical use due to the difference between them.
RESUMEN
The aim of this study was to evaluate the effects of the LL-37, HNP-1 and HBD2/3 peptides on cytokine and MMP production in human polymorphonuclear cells, mononuclear cells and chondrocytes. The levels of cytokines in supernatants from mononuclear and polymorphonuclear cell cultures were measured with a cytometric bead array by flow cytometry. Likewise, the levels of metalloproteinase/MMP-1, 3, and 13 were measured in supernatants from chondrocyte cultures using an ELISA. The expression of RANKL on lymphocytes was analyzed by flow cytometry. We observed increased levels of TNF-α, IL-6 and IL-10 in mononuclear and polymorphonuclear cell cultures stimulated with HBD-2/3. We also observed increased levels of IFN-γ, IL-10, and IL-6 in mononuclear cell cultures stimulated with HNP-1, and increased IL-6 levels were observed in polymorphonuclear cell cultures exposed to HNP-1. We also found that the MMP-1 level increased in the chondrocyte cultures stimulated with HBD-3, whereas the MMP-1 level was decreased in cultures exposed to LL-37. The present report is the first study to determine that HNP-1and HBD2/3 promote the secretion of pro-inflammatory cytokines by polymorphonuclear and mononuclear cells and the secretion of MMP by chondrocytes, whereas LL-37 diminishes MMP1 secretion. Our results suggest that HBD-2/3 and HNP1 might play a pathological role in rheumatoid arthritis, while LL-37 might have a protective role.
Asunto(s)
Catelicidinas/metabolismo , Citocinas/biosíntesis , alfa-Defensinas/metabolismo , beta-Defensinas/metabolismo , Péptidos Catiónicos Antimicrobianos , Condrocitos/metabolismo , Citocinas/metabolismo , Humanos , Leucocitos Mononucleares/metabolismo , Metaloproteinasas de la Matriz/metabolismo , Cultivo Primario de CélulasRESUMEN
AIM: Polymorphisms in CYP2D6 impact the interindividual and interethnic variability of drug efficiency; therefore, we determined the CYP2D6 allele distribution in eight Amerindian groups from northwestern Mexico and compared them with the frequencies in Mexican Mestizos. MATERIALS & METHODS: A total of 508 Amerindians were studied. Genotyping of CYP2D6*5 and multiplication alleles was performed by long-range PCR, while CYP2D6*2, *3, *4, *6, *10, *17, *29, *35, *41 and copy number were evaluated by real-time PCR. RESULTS: The most frequent alleles were CYP2D6*2 (0.05-0.28), CYP2D6*4 (0.003-0.21) and multiplications (0.043-0.107). CYP2D6*5, *6, * 10 and *41 were not observed in the majority of Amerindians, and CYP2D6*3, *17, *35 and *29 were not detected. The poor metabolizer genotype ( *4/*5) was lower (0.2%) in Amerindians than in Mestizos (5%); conversely, the ultrarapid metabolizer genotype was higher (12.6%) in indigenous groups than in Mestizos (7%). CONCLUSION: Our data show a lower frequency of CYP2D6 inactive alleles and a higher frequency of duplication/multiplication of CYP2D6 active alleles in indigenous populations that in Mestizos. Original submitted 14 August 2013; Revision submitted 7 October 2013.
Asunto(s)
Citocromo P-450 CYP2D6/genética , Frecuencia de los Genes , Inactivación Metabólica/genética , Indígenas Norteamericanos/genética , Alelos , Citocromo P-450 CYP2D6/metabolismo , Genotipo , Humanos , México , FenotipoRESUMEN
BACKGROUND: The olfactomedin-like domain (OLFML) is present in at least four families of proteins, including OLFML2A and OLFML2B, which are expressed in adult rat retina cells. However, no expression of their orthologous has ever been reported in human and baboon. OBJECTIVE: The aim of this study was to investigate the expression of OLFML2A and OLFML2B in ocular tissues of baboons (Papio hamadryas) and humans, as a key to elucidate OLFML function in eye physiology. METHODS: OLFML2A and OLFML2B cDNA detection in ocular tissues of these species was performed by RT-PCR. The amplicons were cloned and sequenced, phylogenetically analyzed and their proteins products were confirmed by immunofluorescence assays. RESULTS: OLFML2A and OLFML2B transcripts were found in human cornea, lens and retina and in baboon cornea, lens, iris and retina. The baboon OLFML2A and OLFML2B ORF sequences have 96% similarity with their human's orthologous. OLFML2A and OLFML2B evolution fits the hypothesis of purifying selection. Phylogenetic analysis shows clear orthology in OLFML2A genes, while OLFML2B orthology is not clear. CONCLUSIONS: Expression of OLFML2A and OLFML2B in human and baboon ocular tissues, including their high similarity, make the baboon a powerful model to deduce the physiological and/or metabolic function of these proteins in the eye.