Protective effects of melatonin on changes occurring in the experimental autoimmune encephalomyelitis model of multiple sclerosis.

BACKGROUND: Melatonin has been related to the pathophysiology of multiple sclerosis (MS), and its anti-inflammatory and immunomodulatory properties have been proved in numerous neurodegenerative diseases. This study aimed to find out whether a melatonin supplement in MS is able to act as a benefit to its clinical status, i.e. oxidative stress, inflammation and indirect biomarkers of bacterial dysbiosis, lipopolysaccharide (LPS) and LPS-binding protein (LBP), verifying its therapeutic potential and its possible clinical use in patients with MS. METHODS: The animal MS model, experimental autoimmune encephalomyelitis (EAE), was employed whereby 25 male Dark Agouti rats (5 animals per group) were divided into: a control group (not manipulated); a control+vehicle group; a control+melatonin group; an EAE group; an EAE+melatonin group. Melatonin was administered daily for 51 days, at a dose of 1 mg/kg body weight/i.p., once a day, five days a week. RESULTS: The results from the administration of melatonin demonstrated an improvement in clinical status, a diminution in oxidative stress and inflammation, as well as in bacterial dysbiosis. CONCLUSION: Melatonin could play an effective role against MS, either alone or as a therapy combined with traditional agents.

Helios-Negative Regulatory T Cells as a Key Factor of Immune Tolerance in Nonallergic Beekeepers.

BACKGROUND AND OBJECTIVES: Although exposure to stings has been identified as the leading risk factor for anaphylaxis due to Hymenoptera venom allergy, professional beekeepers receive hundreds of stings yearly without developing systemic reactions. This study aims to analyze the mechanisms underlying bee venom tolerance in beekeepers. METHODS: A cross-sectional study was conducted. Participants were recruited and classified into 3 groups: allergic patients (APs), who experienced systemic reactions after bee stings, with a positive intradermal test and specific IgE (sIgE) to Apis mellifera venom (AmV); tolerant beekeepers (TBKs), who received ≥50 stings/year; and healthy nonexposed controls (HCs). We measured serum levels of sIgE and specific IgG4 (sIgG4) to AmV, rApi m 1, rApi m 2, rApi m 3, Api m 4, rApi m 5, and rApi m10, as well as AmV-induced basophil degranulation, percentage of T-cell subsets, regulatory T cells (Treg), and IL-10 production. RESULTS: Compared with TBKs, APs had high levels of sIgE to AmV and all its allergic components (P<.001), together with a high basophil activation rate (P<.001). Conversely, compared with APs, TBKs had higher levels of sIgG4 (P<.001) and IL-10 (P<.0001), as well as an enhanced CTLA-4+ Treg population (P=.001), expanded Helios- Treg (P<.003), and reduced type 1 helper T cells (TH1) (P=.008), TH2 (P=.004), and TH17 (P=.007) subsets. CONCLUSIONS: The profile of TBKs, which was strongly marked by Treg activity, differed from that of TBKs. This natural tolerance would be led by the expansion of inducible Helios- Treg cells at the peripheral level. The Helios- Treg population could be a novel candidate biomarker for monitoring tolerance.

Helios-Negative Regulatory T Cells as a Key Factor of Immune Tolerance in Nonallergic Beekeepers

Background: Although exposure to stings has been identified as the leading risk factor for anaphylaxis due to Hymenoptera venom allergy, professional beekeepers receive hundreds of stings yearly without developing systemic reactions. Objective: This study aims to analyze the mechanisms underlying bee venom tolerance in beekeepers. Methods: A cross-sectional study was conducted. Participants were recruited and classified into 3 groups: allergic patients (APs), who experienced systemic reactions after bee stings, with a positive intradermal test and specific IgE (sIgE) to Apis mellifera venom (AmV); tolerant beekeepers (TBKs), who received ≥50 stings/year; and healthy nonexposed controls (HCs). We measured serum levels of sIgE and specific IgG4 (sIgG4) to AmV, rApi m 1, rApi m 2, rApi m 3, Api m 4, rApi m 5, and rApi m10, as well as AmV-induced basophil degranulation, percentage of T-cell subsets, regulatory T cells (Treg), and IL-10 production. Results: Compared with TBKs, APs had high levels of sIgE to AmV and all its allergic components (P<.001), together with a high basophil activation rate (P<.001). Conversely, compared with APs, TBKs had higher levels of sIgG4 (P<.001) and IL-10 (P<.0001), as well as an enhanced CTLA-4+ Treg population (P=.001), expanded Helios– Treg (P<.003), and reduced type 1 helper T cells (TH1) (P=.008), TH2 (P=.004), and TH17 (P=.007) subsets. Conclusions: The profile of TBKs, which was strongly marked by Treg activity, differed from that of TBKs. This natural tolerance would be led by the expansion of inducible Helios– Treg cells at the peripheral level. The Helios– Treg population could be a novel candidate biomarker for monitoring tolerance (AU)

Empiema pleural secundario a cálculo biliar. Una complicación infrecuente / Pleural empyema secondary to gallstone. An infrequent complication

Mujer de 84 años que acude a Urgencias por presentar en la semana previa dolor en costado derecho de características pleuríticas asociado a fiebre tras haberse sometido a colecistectomía dos semanas antes. En la radiografía de tórax presenta derrame pleural, confirmándose en la tomografía computarizada de tórax con contraste y objetivándose derrame pleural loculado, así como una imagen compatible con litiasis biliar. Posteriormente se realiza toracocentesis diagnóstica en la que se obtienen resultados compatible con empiema. Se coloca drenaje pleural con instilación de uroquinasa y antibioterapia empírica, presentando buena evolución. Se establece el diagnóstico de empiema pleural secundario a cálculo biliar intraperitoneal (AU)

An 84-year-old woman come to the emergency department for presenting in the previous week pleuritic right flank pain associated with fever after having undergone cholecystectomy 2 weeks earlier. She presents pleural effusion in the chest X-ray, which was confirmed in the chest computered tomography with contrast, showing loculated pleural effusion, as well as an image compatible with biliary lithiasis. Subsequently, diagnostic thoracentesis is performed and the results were compatible with empyema. Pleural drainage is placed with urokinase instillation and empirical antibiotherapy, presenting good evolution. The diagnosis of pleural empyema secondary to intraperitoneal gallstone is established (AU)

3-Methyl-2-benzothiazolinone hydrazone and 3-dimethylamino benzoic acid as substrates for the development of polyphenoloxidase and phenoloxidase activity by zymograms.

In the present study, a sequential staining process of polyphenoloxidase and phenoloxidase enzymes was designed by the zymography technique. As a first step, electrophoresis was carried out under native conditions, and later, first staining was carried out with a revealing solution of 3-methyl-2-benzothiazoline hydrazone (MBTH)-3-dimethylamino benzoic acid (DMAB) that allowed the visualization of polyphenoloxidase enzymes, and later and using the same gel, we proceeded to the differential staining of phenoloxidase, adding a solution of H2O2. The technique was standardized using commercial enzymes of laccase (T. versicolor) and horseradish. The technique was used to identify polyphenoloxidases (laccases) and phenoloxidases (lignin peroxidase) of crude extracts obtained from the growth of the basidiomycete Lentinus strigosus on Pinus radiata. The technique showed great sensitivity to detect the different enzymatic activities (1.56 Activity Unit/mL minimum) in the same gel without interference between the enzymes and the solutions used. On the other hand, the efficiency of the technique was compared with the substrates that are commonly used for the detection of this type of activities such as 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and guaiacol, observing greater sensitivity and minimal interference, so that the present method will allow in the same gel, and visualize polyphenoloxidase and phenoloxidase activities simultaneously facilitating expression studies.

Tos e infiltrados pulmonares. Un final inesperado / Pulmonary infiltrates and cough. An unexpected end

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Isolation and Selection of Streptomyces Species from Semi-arid Agricultural Soils and Their Potential as Producers of Xylanases and Cellulases.

The Mezquital Valley (MV), Mexico, is a semi-arid region whose main economic activity is agriculture, this zone is characterized by the use of wastewater for crop irrigation. This condition has increased the amount nutrients in soils, organic carbon content and native microorganisms. The Streptomyces species are a group of saprophytic bacteria that represent between 20 and 60% of the total microbial population in soils, capable of producing metabolites of commercial importance. In this work, Streptomyces species were isolated from agricultural soils of the MV and was evaluated the production of endoglucanases (CMCase) and xylanases (Xyl) in Solid-State Cultivation (SSC). From soil samples, 73 possible strains of Streptomyces species were isolated for their ability to produce CMCase and Xyl in SSC. The study also included its characterization by morphological characteristics. Of the isolated microorganisms, 38 strains were selected as strong enzyme producers according to the measurement of the halo generated in plate and by growth on barley straw as only carbon source. Two different sizes of barley straw particle were tested, finding that the greatest enzymatic activity was observed in particle size 12. Three strains of Streptomyces species were chosen which presented the best catalytic capacities, a maximum of 100.69 AU Xyl/gram dry matter (gdm), 82 AU Xyl/gdm and 26.02 AU CMCase/gdm for strains 30, 28 and 12, respectively. The strains were identified by ribosomal gen16s sequence and identified as S. flavogriseus, S. virginiae and S. griseoaurantiacus. It is the first report of endogluconase and xylanolytic activity by S. virginiae isolated from a semi-arid soil.

Use of barley straw as a support for the production of conidiospores of Trichoderma harzianum.

In this work was to evaluate the conidiospores production of Trichoderma harzianum using barley straw as substrate. Four growth conditions were used; washed and unwashed barley straw and washed and unwashed barley straw supplemented with mineral salts. The highest spore production was observed when washed barley straw supplemented with mineral salts with 1.56 × 1010 conidiospores/gram of dry matter (gdm) at 216 h of cultivation was used. The effect of substrate moisture on spore production was studied, three initial moisture levels of the substrate were tested and it was observed that a humidity of 80 % of the substrate improves the production of conidiospores reaching a concentration of 2.35 × 1010 conidiospores/gdm at 136 h. Finally, conidiospores viability was evaluated for 12 months by keeping them on the conidia and substrate, and viability of 71 % of the conidiospores was observed, so this maintenance method is an excellent means of conserving the conidiospores viability.

Biological decolorization of Amaranth dye with Trametes polyzona in an airlift reactor under three airflow regimes.

In the present work, a strain of the basidiomycete fungus Trametes polyzona was used to decolorize the Amaranth dye. The decolorization was carried out in an Airlift reactor with three flow regimes: 1, 2, and 3 vvm. The results showed that the decolorization was a function of the flow regime. The decolorization times for the regimes of 1, 2, and 3 vvm were 30, 25, and 19 days, respectively. The COD (Chemical Oxygen Demand) decreased from 1600 to 72 mg COD/L. The enzymatic activity kinetics of laccase (Lcc), lignin peroxidase (LiP), and manganese peroxidase (MnP) were determined. In all the treatments, the enzyme LiP was expressed during the first 6 days, at which point 80% decolorization was observed, whereas Lcc and MnP enzymes were produced from day 6 until the end of the decolorization process. The effluent generated showed no inhibitory effects on the growth of the algae Nannochloropsis salina. T. polyzona showed great versatility in the decolorization of synthetic effluents containing the Amaranth dye, and the fungus was able to use this dye as its only carbon source starting at the beginning of the process. LiP was the enzyme that contributed the most to the decolorization process, and on average, 95% decreases in color and the COD were observed.

Successful adaptation of bee venom ommunotherapy in a patient monosensitized to api m 10

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Valor pronóstico de la invasión de grasa perirrenal y/o del seno en pacientes con carcinoma de células renales pT3a: estudio de cohorte multicéntrico. Grupo LARCG / Prognostic value of perirenal and/or sinus fat infiltration in patients with pT3a renal cell carcinoma: A multicentre cohort study. LARCG Group

Introducción y objetivos: El objetivo de este estudio es evaluar las tasas de sobrevida global (OS), la sobrevida específica del cáncer (CSS), la sobrevida libre de recaída global (RFS), la sobrevida tiempo libre hasta la recaída local (LRFS) y la sobrevida tiempo libre hasta la recaída a distancia (DRFS), en los pacientes con carcinoma de células renales (CCR) pT3a considerando a la infiltración de grasa (FI) perirrenal y/o sinusal como factores pronósticos. Materiales y métodos: Cohorte retrospectiva de pacientes con CCR pT3a sometidos a cirugía. Los datos se extrajeron de la base de datos LARCG. Se evaluaron variables demográficas, clínicas, patológicas y quirúrgicas. La FI se dividió en 4 grupos (vena, perirrenal, sinusal y ambas grasas). Se realizaron curvas de Kaplan-Meier y regresión de Cox. Resultados: Se incluyeron 293 pacientes. La edad media fue de 61,4 años. La mediana de seguimiento fue de 21 meses (r: 1-194). La CSS, la RFS, el LRFS y el DRFS estimadas a 3 años en el grupo de ambas grasas infiltradas fueron 53,1, 45,1, 58,7 y 51,6 meses, respectivamente, en todos los casos estadísticamente inferiores al resto (p ˂ 0,005). En el análisis multivariable, la infiltración de ambas grasas tuvo un aumento significativo de mortalidad específica, recaída global y local con respecto a infiltración de venas (HR: 4,5, 2,42 y 8,08, respectivamente). El grado de Fuhrman y la infiltración de la pelvis renal fueron predictores independientes de la CSS y la RFS. Conclusiones: La infiltración de ambas grasas renales aumenta el riesgo de recaída global y local en pT3a RCC. Del mismo modo, se asocia con una menor sobrevida específica del cáncer, debiendo considerarse como un factor de mal pronóstico

Introduction and objectives: The objective of this study is to evaluate overall survival (OS), cancer-specific survival (CSS), relapse-free survival, local and distant (LRFS and DRFS, respectively) rates in patients with pT3a renal cell carcinoma (RCC) considering the perirenal and/or sinus fat infiltration (FI) as prognostic factors. Materials and methods: Retrospective cohort of patients with pT3a RCC who underwent radical or partial nephrectomy. The data were extracted from the LARCG (Latin American Renal Cancer Group) database. The demographic, clinical, pathological and surgical variables were evaluated. FI was divided into 4 groups (vein, perirenal, sinus and both fats infiltration). The Kaplan Meier and Cox regression curves were performed. Results: 293 patients were included in the study. The mean age was 61.4 years. The median follow-up was 21 months (r: 1-194). CSS, RFS, LRFS and DRFS estimated at 3 years in the group of both fats’ infiltration were 53.1, 45.1, 58.7 and 51.6 months, respectively, and always statistically lower than the rest (P ˂ 0.005). In the multivariate analysis, the infiltration of both fats significantly increased specific mortality, overall and local relapse with respect to vein infiltration (HR: 4.5, 2.42 and 8.08, respectively). The Fuhrman grade and renal pelvis infiltration were independent predictors of CSS and RFS. Conclusions: Infiltration of both fats increases the risk of overall and local relapse in pT3a RCC. In the same way, it is associated with a lower cancer-specific survival and should be considered as a factor of poor prognosis

Dolor abdominal agudo en la enfermedad de Crohn. ¿Qué prueba de imagen urgente se debe hacer? / Acute abdominal pain in patients with Crohn's disease: what urgent imaging tests should be done?

La enfermedad de Crohn es una patología de etiología autoinmunitaria, con afectación predominante del aparato digestivo. Se trata de una enfermedad que se diagnostica a edad temprana y presenta un curso crónico con reagudizaciones. Estos brotes se suelen manejar de forma similar a un abdomen agudo en los pacientes que acuden a los servicios de urgencias, y no existe un consenso sobre cuáles serían las pruebas de imagen más adecuadas. Debido a esta controversia, se ha decidido realizar una revisión de la bibliografía actual sobre cuáles serían las pruebas de imagen indicadas (ya sea por rendimiento diagnóstico o por menor exposición a radiación ionizante) en el brote agudo de pacientes con enfermedad de Crohn

Crohn's disease is an autoimmune disease that predominantly affects the gastrointestinal tract. Crohn's disease is diagnosed at a young age and runs a chronic course with acute flare-ups. When patients with Crohn's disease present with flare-ups at the emergency department, they are usually managed in a way similar to patients with acute abdomen; there is no consensus about the most appropriate imaging work-up for patients with flare-ups of Crohn's disease. Thus, we decided to review the literature about the imaging tests indicated (whether related to their diagnostic performance or to lower exposure to ionizing radiation) for acute flare-ups in patients with Crohn's disease

Prognostic value of perirenal and/or sinus fat infiltration in patients with pT3a renal cell carcinoma: A multicentre cohort study. LARCG Group. / Valor pronóstico de la invasión de grasa perirrenal y/o del seno en pacientes con carcinoma de células renales pT3a: estudio de cohorte multicéntrico. Grupo LARCG.

INTRODUCTION AND OBJECTIVES: The objective of this study is to evaluate overall survival (OS), cancer-specific survival (CSS), relapse-free survival, local and distant (LRFS and DRFS, respectively) rates in patients with pT3a renal cell carcinoma (RCC) considering the perirenal and/or sinus fat infiltration (FI) as prognostic factors. MATERIALS AND METHODS: Retrospective cohort of patients with pT3a RCC who underwent radical or partial nephrectomy. The data were extracted from the LARCG (Latin American Renal Cancer Group) database. The demographic, clinical, pathological and surgical variables were evaluated. FI was divided into 4 groups (vein, perirenal, sinus and both fats infiltration). The Kaplan Meier and Cox regression curves were performed. RESULTS: 293 patients were included in the study. The mean age was 61.4 years. The median follow-up was 21 months (r: 1-194). CSS, RFS, LRFS and DRFS estimated at 3 years in the group of both fats' infiltration were 53.1, 45.1, 58.7 and 51.6 months, respectively, and always statistically lower than the rest (P˂0.005). In the multivariate analysis, the infiltration of both fats significantly increased specific mortality, overall and local relapse with respect to vein infiltration (HR: 4.5, 2.42 and 8.08, respectively). The Fuhrman grade and renal pelvis infiltration were independent predictors of CSS and RFS. CONCLUSIONS: Infiltration of both fats increases the risk of overall and local relapse in pT3a RCC. In the same way, it is associated with a lower cancer-specific survival and should be considered as a factor of poor prognosis.

Acute abdominal pain in patients with Crohn's disease: what urgent imaging tests should be done? / Dolor abdominal agudo en la enfermedad de Crohn. ¿Qué prueba de imagen urgente se debe hacer?

Crohn's disease is an autoimmune disease that predominantly affects the gastrointestinal tract. Crohn's disease is diagnosed at a young age and runs a chronic course with acute flare-ups. When patients with Crohn's disease present with flare-ups at the emergency department, they are usually managed in a way similar to patients with acute abdomen; there is no consensus about the most appropriate imaging work-up for patients with flare-ups of Crohn's disease. Thus, we decided to review the literature about the imaging tests indicated (whether related to their diagnostic performance or to lower exposure to ionizing radiation) for acute flare-ups in patients with Crohn's disease.

Melanoma cutáneo de la región dorsal. reporte de un caso / Cutaneous melanoma of the dorsal region.a case report

RESUMEN: El melanoma es una neoplasia que se origina de los melanocitos, células derivadas de la cresta neural. En el Ecuador, entre el 2001 y 2005 se diagnosticaron en Quito en promedio: 18 casos anuales de melanoma en hombres y 26 casos en mujeres, con una tasa de incidencia de 2.5 y 3.2 respectivamente, mientras queen Guayaquil, durante el período 2003-2006 fueron reportados 49 casos en total, con una incidencia de 0.9 por cada 100.000 habitantes en hombres y 0.1 en mujeres. Siempre que se pueda es preferible una biopsia escisional, extirpando la lesión entera. La determinación del ganglio centinela junto al nivel de Clark, el espesor de Breslow y la ulceración del tumor, son considerados los factores pronósticos más importantes del melanoma maligno.

SUMMARY: Melanoma is a neoplasm that originates from the melanocytes, cells derived from the neural crest. In Ecuador, between 2001 and 2005 an average of 18 cases in men and 26 cases in women were diagnosed in Quito, with an incidence rate of 2.5 and 3.2 respectively, while in Guayaquil, during the period 2003-2006, 49 cases were reported, with an incidence of 0.9 per 100.000 inhabitants in men and 0.1 in women. The exact etiology is not clear, it is considered a heterogeneous diseasewith multifactorial origin. Whenever possible, an excisional biopsy is preferable, removing the entire lesion. The determination of the sentinel lymph node along with the Clark level, the Breslow thickness and the ulceration of the tumor, are considered the most important prognostic factors of malignant melanoma.