Production and characterization of a monoclonal antibody against the sialidase of Gardnerella vaginalis using a synthetic peptide in a MAP8 format.

Bacterial vaginosis is one of the most frequent vaginal infections. Its main etiological agent is Gardnerella vaginalis, which produces several virulence factors involved in vaginal infection and colonization, in particular, sialidase (SLD), a potential clinical biomarker that participates in immune response modulation and mucus degradation. The main objective of this work was the production and evaluation of a monoclonal antibody against G. vaginalis sialidase and its validation in immunoassays. For immunization of mice, a synthetic multiantigenic peptide was used, and hybridomas were generated. After fusion, hybridomas were evaluated for antibody production and cloned by limited dilution. One clone producing IgG1 was selected and characterized by indirect ELISA, dot blot, and Western blot, and we also tested clinical isolates and HeLa cells infected with G. vaginalis. The results showed that the anti-SLD antibody recognized a single protein of ~90 kDa that correlated with the estimated molecular weight of SLD. In addition, anti-SLD antibody recognized SLD from complete bacteria and from culture supernatants of infected Hela cells. In conclusion, our results showed that the anti-SLD antibody recognized SLD from different sources and could be considered a new tool for the diagnosis of bacterial vaginosis. KEY POINTS: • Anti-sialidase mAb was generated using a synthetic peptide • The mAb recognizes synthetic peptide and intact protein from multiple sources • The antibody was characterized by several immunological methods.

THE ACUTE-PHASE AND HEMOSTATIC RESPONSE IN DROMEDARY CAMELS ( CAMELUS DROMEDARIUS).

Acute-phase reactants indicate inflammation and are increasingly used in veterinary medicine to indicate and to monitor progression of disease. Hemostasis and inflammation have interconnected pathophysiologic pathways and influence each other on different levels. This study established observed normal ranges for acute-phase reactants and for coagulation and thromboelastographic (TEG) parameters in 49 dromedary camels ( Camelus dromedarius) and assessed the response to chronic and acute inflammation. Chronically infected animals suffering from lymph abscessation due to Corynebacterium spp. had significantly higher concentrations of the acute-phase reactants haptoglobin ( P < 0.005) and fibrinogen ( P < 0.013) and an increased clot strength characterized by an increase of the TEG parameters MA ( P < 0.039), representing the maximum amplitude of the clot strengths, and G, the global clot strength ( P < 0.022), compared to healthy animals. When the acute-phase and hemostatic responses of 10 males receiving a gonadotropin-releasing hormone vaccine and of 9 males that were surgically castrated over 7 days were studied, haptoglobin proved to be a minor positive acute-phase protein, with moderate levels in healthy animals. It increased significantly after both vaccination and castration and remained elevated 7 days postinsult. The negative reactant iron significantly decreased over the 7-day period after castration, whereas a similar decrease following vaccination lasted less than 3 days. Fibrinogen reacted as a positive, minor reactant, with a significant increase and a peak on days 3-5, with higher values seen after castration. Prothrombin time showed a slight shortening at days 5-7, and the TEG parameters MA and G showed significantly increased values, similar to fibrinogen. The acute-phase protein serum amyloid A showed poor repeatability, suggesting that the assay was not reliable.

Genetic and phenoptypic differentiation of zebra mussel populations colonizing Spanish river basins.

Zebra mussel populations in Ebro and Mijares Rivers (northern Spain) were analyzed to study the mechanisms by which this aquatic species deals with pollution. Variability analyses of mitochondrial cytochrome oxidase I gene and of one nuclear microsatellite were performed for ten populations from the Ebro River and one from the Mijares River. Comparison of these results with those from five additional European populations indicated that the Spanish populations constitute a homogeneous gene pool. Transcriptome analyses of gill samples from a subset of the Spanish populations showed changes on expression levels that correlated with variations in general fitness and loads of heavy metals. The less polluted upstream Ebro populations showed overexpression of mitochondrial and cell proliferation-related genes compared to the more polluted, downstream Ebro populations. Our data indicate that heavy metals were the main factors explaining these transcriptomic patterns, and that zebra mussel is resilient to pollutants (like mercury and organochlorine compounds) proved to be extremely toxic to vertebrates. We propose that zebra mussel populations sharing a common gene pool may acclimate to different levels and forms of pollution through modulations in their transcriptomic profile, although direct selection on genes showing differential expression patterns cannot be ruled out.

Use of monoclonal antibodies in cancer immunotherapy: types and mechanisms of action.

Immunotherapy is one of the most innovative treatments in the current field of oncology and consists of stimulating the immune system to eliminate tumoral cells. Monoclonal antibodies (mAbs) are glycoproteins secreted by B-cells capable of recognizing and neutralizing foreign organisms or antigens. Structurally, they are composed of two heavy and two light chains. The generation of therapeutic mAbs is one of the most developed and fastest-growing areas of the biotechnological and pharmaceutical industries and is an important adjunct to cancer therapy. Several antibodies have been approved for human administration and can be mouse-derived, chimeric, humanized, or fully human. mAbs main mechanism of action includes the lysis of the tumoral cells through inducing apoptosis, phagocytosis, complement activation, or signaling inhibition.

La inmunoterapia es un tratamiento innovador para la oncología actual, que consiste en la estimulación del sistema inmunitario para la eliminación de las células tumorales. Los anticuerpos monoclonales (mAbs) son glicoproteínas secretadas por los linfocitos B, capaces de reconocer y neutralizar organismos extraños o antígenos. Estructuralmente se componen de dos cadenas pesadas y dos cadenas ligeras. La generación de mAbs terapéuticos es una de las áreas de mayor crecimiento en la industria biotecnológica y farmacéutica y representa un complemento importante en la terapia del cáncer. Existen diversos mAbs que han sido aprobados para su administración en humanos, y pueden ser derivados de ratón, quiméricos, humanizados o completamente humanos. Los mecanismos de acción consisten principalmente en la lisis de las células tumorales a través de la inducción de la apoptosis, fagocitosis, activación del complemento o inhibición de la señalización celular.

Autophagy in Hematological Malignancies.

Autophagy is a highly conserved metabolic pathway via which unwanted intracellular materials, such as unfolded proteins or damaged organelles, are digested. It is activated in response to conditions of oxidative stress or starvation, and is essential for the maintenance of cellular homeostasis and other vital functions, such as differentiation, cell death, and the cell cycle. Therefore, autophagy plays an important role in the initiation and progression of tumors, including hematological malignancies, where damaged autophagy during hematopoiesis can cause malignant transformation and increase cell proliferation. Over the last decade, the importance of autophagy in response to standard pharmacological treatment of hematological tumors has been observed, revealing completely opposite roles depending on the tumor type and stage. Thus, autophagy can promote tumor survival by attenuating the cellular damage caused by drugs and/or stabilizing oncogenic proteins, but can also have an antitumoral effect due to autophagic cell death. Therefore, autophagy-based strategies must depend on the context to create specific and safe combination therapies that could contribute to improved clinical outcomes. In this review, we describe the process of autophagy and its role on hematopoiesis, and we highlight recent research investigating its role as a potential therapeutic target in hematological malignancies. The findings suggest that genetic variants within autophagy-related genes modulate the risk of developing hemopathies, as well as patient survival.

Nanophotonic Sialidase Immunoassay for Bacterial Vaginosis Diagnosis.

Bacterial vaginosis (BV) affects reproductive-age women and can lead to pelvic inflammatory disease, postpartum endometritis, and preterm labor/delivery and predisposes the infection of sexually transmitted diseases. Typically, BV diagnosis involves the analysis of vaginal swab samples via microscopy operated by highly skilled personnel. Hence, novel approaches for BV diagnosis are an existing need. In response, the first immunosensing platform targeting sialidase, a BV biomarker, is reported. The nanophotonic operational principle of this biosensing platform allows for a cheaper, faster, and simpler analysis when compared with an indirect enzyme-linked immunosorbent assay (ELISA). The clinical evaluation of such a nanotechnology is highlighted, where 162 vaginal swab samples were analyzed with high sensitivity and specificity (96.29%, respectively). The resulting nanoimmunosensing platform offers a resourceful approach to perform a timely BV diagnosis.

Changes in lipid profile and insulin resistance in obese patients after Scopinaro biliopancreatic diversion.

BACKGROUND: Obesity is associated with many cardiovascular risk factors. The aim of this study is to evaluate the effect of bariatric surgery (BS) in lipid profile and insulin resistance in obese patients. MATERIAL AND METHODS: We studied changes in lipid parameters glucose, insulin, and insulin resistance (IR) before biliopancreatic diversion and 3, 6, 12, 18, and 24 months after surgery, in 115 obese patients divided in two groups: diabetics (DM) and nondiabetics (non-DM). RESULTS: In both groups, all parameters significantly decreased at 3 months; however, high-density lipoprotein cholesterol levels (HDL-c) and total cholesterol (TC) to HDL-c ratio decreased up to 6 months. At baseline, in DM, we found a negative correlation between body mass index (BMI) and TC to HDL-c ratio and glucose with HDL-c and homeostasis model assessment (HOMA) with BMI and a direct correlation between glucose with TC to HDL-c ratio and glucose with triglycerides. In non-DM, no correlations were found. Comparing both groups for each parameter, we found significant differences at basal levels for glucose, HOMA, triglycerides (TG), and TG to HDL-c ratio. Changes between basal levels and 1 year after surgery were significantly different in glucose and HOMA. CONCLUSION: BS offers excellent results in weight reduction and significantly improves IR, diabetes, and the lipid profile, decreasing global cardiovascular risk.

Production and characterization of monoclonal antibodies against the DNA binding domain of the RE1-silencing transcription factor.

The use of monoclonal antibodies for the detection of cellular biomarkers during carcinogenesis provides new strategies for cancer diagnosis or prognosis in patients. Loss of the Restrictive Element 1-Silencing Transcription (REST) factor has been observed in previous molecular and immunological approaches in aggressive breast cancer, small cell lung cancer, liver carcinoma, and colo-rectal cancer; however, for clinic diagnosis, monoclonal antibodies for REST recognition are unavailable. The goal of this work was to design, produce and characterize monoclonal antibodies against the REST DNA binding damain (DBD) that would be suitable for immunoassays. We searched for conserved domains, and immunogenic and antigenic sites in the REST structure via in silico analysis. For mice immunization, we used a recombinant REST DBD purified by affinity chromatography, and then Hybridomas were generated by mouse spleen fusion with myeloma cells. Finally, for monoclonal antibody characterization, we performed enzyme-linked immunosorbent (ELISA), western blot, dot blot, immunocytochemistry (ICC) and immunoprecipitation assays. Results showed that the DBD is conserved in REST isoforms and contains immunogenic and antigenic sites. We generated three clones producing monoclonal antibodies against REST DBD, one of them specifically recognized native REST and was suitable for ICC in samples from patients.

Use of monoclonal antibodies in cancer immunotherapy: types and mechanisms of action / Uso de anticuerpos monoclonales en la inmunoterapia del cáncer: tipos y mecanismos de acción

Abstract Immunotherapy is one of the most innovative treatments in the current field of oncology and consists of stimulating the immune system to eliminate tumoral cells. Monoclonal antibodies (mAbs) are glycoproteins secreted by B-cells capable of recognizing and neutralizing foreign organisms or antigens. Structurally, they are composed of two heavy and two light chains. The generation of therapeutic mAbs is one of the most developed and fastest-growing areas of the biotechnological and pharmaceutical industries and is an important adjunct to cancer therapy. Several antibodies have been approved for human administration and can be mouse-derived, chimeric, humanized, or fully human. mAbs main mechanism of action includes the lysis of the tumoral cells through inducing apoptosis, phagocytosis, complement activation, or signaling inhibition.

Resumen La inmunoterapia es un tratamiento innovador para la oncología actual, que consiste en la estimulación del sistema inmunitario para la eliminación de las células tumorales. Los anticuerpos monoclonales (mAbs) son glicoproteínas secretadas por los linfocitos B, capaces de reconocer y neutralizar organismos extraños o antígenos. Estructuralmente se componen de dos cadenas pesadas y dos cadenas ligeras. La generación de mAbs terapéuticos es una de las áreas de mayor crecimiento en la industria biotecnológica y farmacéutica y representa un complemento importante en la terapia del cáncer. Existen diversos mAbs que han sido aprobados para su administración en humanos, y pueden ser derivados de ratón, quiméricos, humanizados o completamente humanos. Los mecanismos de acción consisten principalmente en la lisis de las células tumorales a través de la inducción de la apoptosis, fagocitosis, activación del complemento o inhibición de la señalización celular.

Formación posgraduada y modos hegemônicos de análisis de políticas públicas

Objetivo Caracterizar los modos de enseñar e investigar políticas públicas a nivel de posgrado en Colombia. Métodos Investigación cualitativa de tipo análisis documental. Inicialmente, se realiza una caracterización general de autores, enfoques teóricos empleados, temáticas de interés, entre otros. En un segundo momento, y a partir del contenido de los programas académicos de los posgrados de donde emergieron las tesis analizadas, se identificaron significantes centrales del discurso curricular. Resultados Se analizaron 313 tesis y 8 programas de posgrado en políticas públicas. Se destaca una marcada preferencia por el empleo de enfoques pospositivistas y estrategias tipo problemsolving; cuestionamientos débiles a determinantes estructurales de opresión, exclusión y explotación, y mantenimiento de relaciones de dominación en la toma de decisiones entre investigadores de ciencias políticas/administrativas y sujetos con otros saberes. Conclusiones Los hallazgos del estudio problematizan el campo del Análisis de Políticas Públicas, sus posturas epistemológicas y métodos predominantes, así como el papel que asume el analista, invitando al lector a reflexionar sobre la necesidad de incursionar en enfoques críticos alternativos.

Objective To describe the methods used at a postgraduate level to teach and research public policies in Colombia. Methods Documentary analysis as part of a qualitative study. Among other things, a broad characterisation of authors, theoretical techniques employed, and areas of interest was accomplished in a first instance. Then, the main significants of the programmatic discourse were recognized based on the postgraduate curriculum from the universities from which the investigated thesis came. Results 313 theses and 8 postgraduate programs in public policy analysis were studied. There is a clear preference for post-positivist mainstream approaches and problem-solving strategies; weak challenges to structural determinants of oppression, exclusion, and exploitation; and the preservation of dominant relationships in decision-making between political/administrative science researchers and subjects with other knowledge. Conclusions The research's findings challenge the discipline of Public Policy Analysis, its epistemological views, and its dominant study methodologies, as well as the analyst's role, encouraging the reader to consider the need for alternative critical approaches.

Macrocytic anemia and thrombocytopenia induced by orlistat.

INTRODUCTION: The overall incidence of obesity and its prevalence is increasing continuously. The obesity is a cardiovascular risk factor whose importance is increasing too. It is associated with many chronic conditions such as type II diabetes mellitus or cardiovascular diseases. The obesity is also implicated as a risk factor for several kinds of cancer such as esophagus, pancreas, colon, rectum, breast cancer in menopausal women. The treatment of the obesity may reduce the incidence of these diseases. The mainstray of the treatment of obesity is changing the lifestyles, but obesity´s treatment may need drug therapy or even though surgical treatment. Orlistat is a specific inhibitor of gastrointestinal lipases, which stops fat absortion. It is used along with a hypocaloric diet, for obesity´s treatment. The beneficial effects of orlistat include weight loss, the improvement of blood pressure´s control, it may delay the development of diabetes mellitus, and it may reduce HbA1c. CASE REPORT: Besides the interaction with other drugs (mainly warfarin and amiodarone). Orlistat´s mainly side effects are gastrointestinal disorders such as the existence of oily spotting from the rectum, abdominal pain or discomfort, fecal urgency. There are also side effects at other levels, like flu symptoms, hypoglycemia, heathache or upper respiratory infections. There are other side effects with very low incidence but clinically relevant like pancreatitis, subacute liver failure, severe liver disease, myopathy, or tubular necrosis secondary to oxalate nephropathy induced by Orlistat. DISCUSSION: IN THIS CASE REPORT APPEARS A NEW ADVERSE EFFECT OF ORLISTAT THAT HAS NOT BEEN DESCRIBED ABOVE: thrombopenia and macrocytic anemia.

Comportamiento del tabaquismo en los adolescentes del Reparto Fructuoso Rodríguez, Cárdenas / Behavior of smoking in teenagers of the Fructuoso Rodriguez Neighborhood. Cardenas

Introducción: la Organización Mundial de la Salud reporta al tabaquismo como la principal causa prevenible de enfermedad, discapacidad y muerte prematura, y ha señalado que mueren aproximadamente 14 000 personas al día, por alguna de las enfermedades atribuibles a este. Objetivo: caracterizar el comportamiento del tabaquismo en adolescentes. Materiales y métodos: la investigación fue observacional, descriptiva, transversal, en un universo de 51 adolescentes de los consultorios 24, 25 y 26 del Policlínico Universitario “Héroes del Moncada”, de Cárdenas, que cumplieron los siguientes criterios de inclusión: tener edad comprendida entre 10 y 19 años, ser fumador activo, independientemente del tiempo con el hábito, la cantidad de cigarrillos que fume, y con consentimiento asistido por los padres de participar en el estudio durante el período de enero a junio de 2013. Resultados: el 11,4 % de los adolescentes son fumadores, y del sexo masculino 78,4 %. Predominó el grupo de 17 a 19 años, con un 58,8 %. El 92,2 % comenzó a fumar a los 12 años, el 56,9 % lo hace diariamente, y compran personalmente el cigarro en las tiendas el 43,8 %. Todos fuman públicamente, y en casa lo hace el 62,7 %. Conclusiones: predominaron los fumadores varones entre 17 y 19 años de edad. La mayoría de los adolescentes fuman diariamente en lugares públicos, compran personalmente los cigarros en las tiendas y comenzaron a fumar con más de 12 años de edad.

Background: the World Health Organization reports smoking as the main preventable cause of disease, disability and premature death, and has pointed out that around 14 000 persons per day die due to any of the diseases attributed to it. Aim: characterizing smoking behavior in teenagers. Material and Methods: the research was observational, descriptive, cross-sectional, in a universe of 51 teenagers belonging to the family physician consultations 24, 25 and 26 of the Teaching Policlinic “Heroes del Moncada”, of Cardenas, fulfilling the following inclusion criteria: aged 10-19 years, being active smokers without taking into account the time they have being smoking, the quantity of cigarettes they smoke, and having their relatives´ informed consent for participating in the research in the period from January to June 2013. Outcomes: 11.4 % of the teenagers were smokers, and 78.4 % were male. There it was a predominance of the 17-19 years-old group, for 58.8 %. 92.2 % of them began smoking at the age of 12, 56.9 percent smokes every day, and 43.8 % buys cigarettes in the stores by themselves. All of them smoke in public, and 62.7 % smokes at home. Conclusions: there it is a predominance of male teenagers aged 17-19 years. Most of the teenagers smoke every day in public places, buy personally cigarettes in the stores and began smoking when they were more than 12 years old.

Effects of dietary L-arginine intake on cardiorespiratory and metabolic adaptation in athletes.

To assess the effect of diet enrichment with L-arginine or supplementation at high doses on physiological adaptation during exercise, 9 athletes followed 3 different diets, each over 3 consecutive days, with a wash-out period of 4 d between training sessions: control diet (CD), 5.5 +/- 0.3 g/d of L-arginine; Diet 1 (rich in L-arginine food), 9.0 +/- 1.1 g/d of L-arginine; and Diet 2 (the same as CD but including an oral supplement of 15 g/d), 20.5 +/- 0.3 g/d of L-arginine. Plasma nitrate levels of each participant were determined on the day after each treatment. Participants performed a submaximal treadmill test (initial speed 10-11 km/hr, work increments 1 km/hr every 4 min until 85-90% VO2max, and passive recovery periods of 2 min). Oxygen uptake and heart rate were monitored throughout the test. Blood lactate concentration ([La-]b) was determined at the end of each stage. Repeated-measures ANOVA and paired Student's t tests were used to compare the various physiological parameters between diets. The level of significance was set at p < .05. [La-]b showed a significant effect at the 5-min time point between CD and Diet 2 (CD 3.0 +/- 0.5 mM, Diet 2 2.5 +/- 0.5 mM, p = .03), but this tendency was not found at higher exercise intensities. No significant differences were observed in any of the cardiorespiratory or plasma nitrate levels. In conclusion, dietary L-arginine intake on the days preceding the test does not improve physiological parameters during exercise.

Representaciones sociales de la inclusión escolar de estudiantes con limitación visual: creencias y prácticas / Social representation of school inclusion students with vision impairment: Beliefs and practices

Esta investigación tuvo como objetivo comprender las Representaciones Sociales que tienen los diferentes actores de la comunidad educativa de 10 colegios públicos de Bogotá, frente a las experiencias de Inclusión Escolar de estudiantes con limitación visual. Las representaciones sociales son definidas como maneras espe-cíficas de entender y comunicar la realidad. Se realizo un estudio de enfoque cualitativo, hermenéutico- inter-pretativo; la estrategia de Investigación fue el estudio de caso y la técnica de recolección de información fue la entrevista de grupo focal. Se realizó un Análisis de Dominio (Spradley, 1979) que permitió comprender las representaciones sociales presentes en el discurso de los participantes. La información obtenida fue analizada a partir de las siguientes categorías: Necesidades y Expectativas, Prácticas y Significado de la discapacidad. Los resultados de esta investigación permiten concluir que si bien, existen contenidos compartidos en algunas de las representaciones sociales que han construido los diferentes actores de la comunidad educativa en relación con los procesos de inclusión escolar de estudiantes con limitación visual, también evidencian diferencias entre ellos a partir de su rol, experiencia y relación con esta condición. Se generó conocimiento en relación con las creencias y prácticas educativas que tienen los diferentes actores involucrados en las experiencias de inclusión escolar de personas con limitación visual.

The aim of this research study was to understand the social representations of different stakeholders at 10 Bogota's public schools, related with students' experiences all of them with visual impairment. Nowadays, Social representations are defined as specific ways to understand and communicate the reality. We realized a qualitative study approach which research strategy was the case study and data collection technique was the focus group interview. We used adomainanalysis(Spradley, 1979)by understand the social representation-sin participant's discourse. The information obtained wasanalyzed fromthe following categories: Needs and Expectations, Practices andMeaningof Disability. By our results we concluded that in spitethere aresomesha-red contentin social representations there are somedifferences between different stakeholders associated with theirrole, experience and relationship with this condition. New knowledge was generated about educational practices and beliefs related with differents takeholders involved in the experiences of educational inclusion of students with visual impairment

Laboratory diagnosis of human neurocysticercosis: double-blind comparison of enzyme-linked immunosorbent assay and electroimmunotransfer blot assay.

Neurocysticercosis is a common disease in underdeveloped countries. Its diagnosis is based on clinical, imaging (tomography or magnetic resonance), epidemiological, and laboratory data. Several methods based on the detection of antibodies against cysticerci in cerebrospinal fluid or serum have been tested. Among them, an enzyme-linked immunosorbent assay (ELISA) based on the use of a crude parasite antigen has been used by the laboratory network of cysticercosis in Mexico, which has given support to clinicians for up to 7 years. A Taenia solium-specific glycoprotein-based electroimmunotransfer blot (EITB) assay was reported to be highly sensitive and specific for this purpose. In order to compare both techniques, we studied 100 neurocysticercosis patients and 70 neurological noncysticercosis controls and searched for specific antibodies in paired samples of serum and cerebrospinal fluid using both techniques. We found that the EITB assay is more sensitive than the ELISA, especially when serum is being tested. Both techniques are more sensitive in cases with multiple living cysts than in cases with single cysts or calcified lesions. No global differences among cases with parasites located in different parts of the central nervous system were found. In the patients with cysts within the parenchyma, the sensitivity of the EITB assay was higher with serum than with cerebrospinal fluid. The immunodominant bands were found to be the same as those previously reported, i.e., GP-39 to -42, GP-24, and GP-13. Based on these results, we suggest the use of the EITB assay in routine diagnosis of cysticercosis for clinical cases.