Ascorbyl palmitate (ASC16) as a potential inhibitor of toxicity induced by Crotalus durissus terrificus venom.

It is well known that C. d. terrificus venom causes pathophysiological effects such as neuropathies, coagulopathies, and even death. Previous studies have reported that ASC16 can interact with monomeric phospholipases A2 from the venom of various snake species (e.g., Vipera russelli and Echis carinatus). As a result, ASC16 has been proposed as an inhibitor of the toxic effects induced by the heterodimeric complex (crotoxin) and other components of the venom of C. d. terrificus. To investigate this further, in silico studies were designed using the crotoxin (CTX) protein complex as a model, and experimental assays were conducted to evaluate the inhibitory effect of ASC16 on CTX, as well as on other venom enzymes such as thrombin-like enzyme (TLE), phosphodiesterase (PDE) and l-aminoxidase (LAAO). For in vitro assays, specific substrates were used, and lethal activity was measured over 48 h using an in vivo murine experimental model (CF01). In silico studies have indicated that the hydrophilic portion of ASC16 adopts a stable conformation while interacting with the catalytic site of crotoxin. At the highest concentrations, ASC16 significantly inhibited the activities of PLA2 (40.89 ± 0.09 %), TLE (11.03 ± 0.69 %), PDE (51.33 ± 2.83 %), and LAAO (56.79 ± 2.91 %). Furthermore, ASC16 neutralized the 2 LD50 lethality of crotalic venom. These findings lay the groundwork for designing promising adjuvants that can facilitate the incorporation of a larger quantity of proteins in immunization schemes. Consequently, this approach aims to achieve higher antibody titers, reduce the number of required immunizations, and minimize local damage in the producer animal.

Fast venomic analysis of Crotalus durissus terrificus from northeastern Argentina

The complete knowledge of the toxins that make up venoms is the base for the treatment of snake accidents victims and the selection of specimens for the preparation of venom pools for antivenom production. In this work, we used a fast and direct venomics approach to identify the toxin families in the C.d. terrificus venom, a Southern American Neotropical rattlesnake. The RP-HPLC separation profile of pooled venom from adult specimens followed by mass spectrometry analysis revealed that C.d. terrificus’ venom proteome is composed of 12 protein families, which are unevenly distributed in the venom, e.g., there are few major proteins in the venom's composition phospholipase A2, serine proteinase, crotamine and L-amino acid oxidase. At the same time, the proteome analysis revealed a small set of proteins with low quantity (less than 1.5%), both enzymes (metaloprotease, phospholipase B and 5′-nucleotidase) and proteins (Bradykinin potentiating and C-type natriuretic peptides, C-type lectin convulxin and nerve growth factor). To sum up, this research is the first venomic report of C.d.terrificus venom from Argentina. This proved to be crotamine positive venom that has a lower metalloprotease content than C.d. terrificus venoms from other regions. This information could be used in the discovery of future pharmacological agents or targets in antivenom therapy.

Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae)

Background: Argenteohyla siemersi (red-spotted Argentina frog) is a casque-headed tree frog species belonging to the Hylidae family. This species has a complex combination of anti-predator defense mechanisms that include a highly lethal skin secretion. However, biochemical composition and biological effects of this secretion have not yet been studied. Methods: The A. siemersi skin secretion samples were analyzed by mass spectrometry and chromatographic analysis (MALDI-TOF/MS, RP-HPLC and GC-MS). Proteins were also studied by SDS-PAGE. Among the biological activities evaluated, several enzymatic activities (hemolytic, phospholipase A2, clotting, proteolytic and amidolytic) were assessed. Furthermore, the cytotoxic activity (cytolysis and fluorescence staining) was evaluated on myoblasts of the C2C12 cell line. Results: The MALDI-TOF/MS analysis identified polypeptides and proteins in the aqueous solution of A. siemersi skin secretion. SDS-PAGE revealed the presence of proteins with molecular masses from 15 to 55 kDa. Steroids, but no alkaloids or peptides (less than 5 KDa), were detected using mass spectrometry. Skin secretion revealed the presence of lipids in methanolic extract, as analyzed by CG-MS. This secretion showed hemolytic and phospholipase A2 activities, but was devoid of amidolytic, proteolytic or clotting activities. Moreover, dose-dependent cytotoxicity in cultured C2C12 myoblasts of the skin secretion was demonstrated. Morphological analysis, quantification of lactate dehydrogenase release and fluorescence staining indicated that the cell death triggered by this secretion involved necrosis. Conclusions: Results presented herein evidence the biochemical composition and biological effects of A. siemersi skin secretion and contribute to the knowledge on the defense mechanisms of casque-headed frogs.

Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae)

Background: Argenteohyla siemersi (red-spotted Argentina frog) is a casque-headed tree frog species belonging to the Hylidae family. This species has a complex combination of anti-predator defense mechanisms that include a highly lethal skin secretion. However, biochemical composition and biological effects of this secretion have not yet been studied. Methods: The A. siemersi skin secretion samples were analyzed by mass spectrometry and chromatographic analysis (MALDI-TOF/MS, RP-HPLC and GC-MS). Proteins were also studied by SDS-PAGE. Among the biological activities evaluated, several enzymatic activities (hemolytic, phospholipase A2, clotting, proteolytic and amidolytic) were assessed. Furthermore, the cytotoxic activity (cytolysis and fluorescence staining) was evaluated on myoblasts of the C2C12 cell line. Results: The MALDI-TOF/MS analysis identified polypeptides and proteins in the aqueous solution of A. siemersi skin secretion. SDS-PAGE revealed the presence of proteins with molecular masses from 15 to 55 kDa. Steroids, but no alkaloids or peptides (less than 5 KDa), were detected using mass spectrometry. Skin secretion revealed the presence of lipids in methanolic extract, as analyzed by CG-MS. This secretion showed hemolytic and phospholipase A2 activities, but was devoid of amidolytic, proteolytic or clotting activities. Moreover, dose-dependent cytotoxicity in cultured C2C12 myoblasts of the skin secretion was demonstrated. Morphological analysis, quantification of lactate dehydrogenase release and fluorescence staining indicated that the cell death triggered by this secretion involved necrosis. Conclusions: Results presented herein evidence the biochemical composition and biological effects of A. siemersi skin secretion and contribute to the knowledge on the defense mechanisms of casque-headed frogs.

Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae)

Argenteohyla siemersi (red-spotted Argentina frog) is a casque-headed tree frog species belonging to the Hylidae family. This species has a complex combination of anti-predator defense mechanisms that include a highly lethal skin secretion. However, biochemical composition and biological effects of this secretion have not yet been studied. Methods: The A. siemersi skin secretion samples were analyzed by mass spectrometry and chromatographic analysis (MALDI-TOF/MS, RP-HPLC and GC-MS). Proteins were also studied by SDS-PAGE. Among the biological activities evaluated, several enzymatic activities (hemolytic, phospholipase A2, clotting, proteolytic and amidolytic) were assessed. Furthermore, the cytotoxic activity (cytolysis and fluorescence staining) was evaluated on myoblasts of the C2C12 cell line. Results: The MALDI-TOF/MS analysis identified polypeptides and proteins in the aqueous solution of A. siemersi skin secretion. SDS-PAGE revealed the presence of proteins with molecular masses from 15 to 55 kDa. Steroids, but no alkaloids or peptides (less than 5 KDa), were detected using mass spectrometry. Skin secretion revealed the presence of lipids in methanolic extract, as analyzed by CG-MS. This secretion showed hemolytic and phospholipase A2 activities, but was devoid of amidolytic, proteolytic or clotting activities. Moreover, dose-dependent cytotoxicity in cultured C2C12 myoblasts of the skin secretion was demonstrated. Morphological analysis, quantification of lactate dehydrogenase release and fluorescence staining indicated that the cell death triggered by this secretion involved necrosis. Conclusions: Results presented herein evidence the biochemical composition and biological effects of A. siemersi skin secretion and contribute to the knowledge on the defense mechanisms of casque-headed frogs.(AU)

Evaluation of pro-inflammatory events induced by Bothrops alternatus snake venom

Inflammation is a major local feature of envenomation by bothropic snakes being characterized by a prominent local edema, pain, and extensive swelling. There are reports demonstrating that whole Bothrops snake venoms and toxins isolated from them are able to activate macrophages functions, such as phagocytosis, production of reactive oxygen, cytokines and eicosanoids, however, little is known about the effects of Bothrops alternatus (B.alpha.) venom on macrophages. In this work, we evaluated the proinflammatory effects of B.alpha. venom with in vivo and in vitro experiments using the Raw 264.7 cell line and mouse peritoneal macrophages. We detected that B.alpha. venom augments cell permeability (2-fold), and cellular extravasation (mainly neutrophils), increase proinflammatory cytokines IL1 (similar to 300-fold), IL12 (similar to 200-fold), and TNF alpha (similar to 80-fold) liberation and induce the expression of enzymes related to lipid signaling, such as cPLA(2 alpha) and COX-2. Additionally, using lipidomic techniques we detected that this venom produces a release of arachidonic acid (similar to 10 nMol/mg. Protein) and other fatty acids (16:0 and 18:1 n-9c). Although much of these findings were described in inflammatory processes induced by other bothropic venoms, here we demonstrate that B.alpha. venom also stimulates pro-inflammatory pathways involving lipid mediators of cell signaling. In this sense, lipidomics analysis of macrophages stimulated with B.alpha. venom evidenced that the main free fatty acids are implicated in the inflammatory response, and also demonstrated that this venom, is able to activate lipid metabolism even with a low content of PLA(2).

Evaluation of pro-inflammatory events induced by Bothrops alternatus snake venom

Inflammation is a major local feature of envenomation by bothropic snakes being characterized by a prominent local edema, pain, and extensive swelling. There are reports demonstrating that whole Bothrops snake venoms and toxins isolated from them are able to activate macrophages functions, such as phagocytosis, production of reactive oxygen, cytokines and eicosanoids, however, little is known about the effects of Bothrops alternatus (B.alpha.) venom on macrophages. In this work, we evaluated the proinflammatory effects of B.alpha. venom with in vivo and in vitro experiments using the Raw 264.7 cell line and mouse peritoneal macrophages. We detected that B.alpha. venom augments cell permeability (2-fold), and cellular extravasation (mainly neutrophils), increase proinflammatory cytokines IL1 (similar to 300-fold), IL12 (similar to 200-fold), and TNF alpha (similar to 80-fold) liberation and induce the expression of enzymes related to lipid signaling, such as cPLA(2 alpha) and COX-2. Additionally, using lipidomic techniques we detected that this venom produces a release of arachidonic acid (similar to 10 nMol/mg. Protein) and other fatty acids (16:0 and 18:1 n-9c). Although much of these findings were described in inflammatory processes induced by other bothropic venoms, here we demonstrate that B.alpha. venom also stimulates pro-inflammatory pathways involving lipid mediators of cell signaling. In this sense, lipidomics analysis of macrophages stimulated with B.alpha. venom evidenced that the main free fatty acids are implicated in the inflammatory response, and also demonstrated that this venom, is able to activate lipid metabolism even with a low content of PLA(2).

Factors related to total energy expenditure in older adults (Chile) / Factores relacionados con el gasto energético total en adultos mayores (Chile)

Aim: to asses Total Energy Expenditure (TEE) in healthy Chilean institutionalized or independently older people Methods: twenty seven young (27-30 years), 27 institutionalized (> 65 years old) and 27 free-living older (> 65 years old) participants were studied. Body composition was estimated by dual energy X-ray absorptiometry. Physical activity energy expenditure (AEE) and TEE were assessed using Actiheart accelerometers. The Mini Nutritional Assessment (MNA) was applied and Timed Up and Go (TUG) was measured. Results: AEE was 171, 320 and 497 kcal/day in institutionalized, free living older and young participants, respectively (p< 0.01 between young and older participants). Both absolute TEE and TEE/RMR was higher in young people. Multiple regression analysis accepted age, MNA and TUG as significant predictors of AEE (r2 = 0.24 p< 0.01). Conclusion: AEE and PAL were lower among older people, with no differences by institutionalization (AU)

Objetivo: evaluar el Gasto Energético Total (GET) en ancianos sanos que viven institucionalizados o independientes en Chile. Método: se evaluaron veintisiete jóvenes (27-30 años), 27 adultos mayores institucionalizados (> 65 años) y 27 ancianos independientes (> 65 años). Se midió la composición corporal utilizando absorciometría bifotónica de rayos X. Se calculó el gasto energético por actividad física (GEAF) y el gasto energético total (GET) utilizando acelerómetros Actiheart; se aplicó Mini Nutritional Assessment (MNA) y se midió el Timed Up and Go (TUG). Resultados: el GEAF fue 171, 320 y 497 kcal/día en ancianos institucionalizados, independientes y jóvenes, respectivamente (p <0,01 entre jóvenes y ancianos). Tanto el valor absoluto de GET como GET/GER fue más alto entre los jóvenes. El análisis de regresión múltiple aceptó la edad, el MNA y el TUG como predictores significativos de GEA (r2 = 0,24 p < 0,01). Conclusión: el GEAF y el nivel de actividad física (PAL) fueron más bajos en los adultos mayores, sin diferencias por institucionalización (AU)

Central obesity and not age increases skeletal muscle lipids, without influencing lean body mass and strength / La obesidad central y no la edad aumenta la grasa intramiocelular, sin afectar la masa y funcion muscular

Background/Aims: To measure skeletal muscle lipid infiltration, its association with insulin resistance (IR) lean mass and function, in Chilean men differing in age and body composition. Our hypothesis was that muscle lipid accumulation would be higher among older and heavier individuals and this would deteriorate insulin sensitivity (IS) and decrease muscle mass and function, both features of the ageing process. Methods: Healthy men (38 < 55 and 18 > 65 years), underwent anthropometric measurements, body composition assessment through radiologic densitometry, Nuclear Magnetic Resonance spectroscopy at the tibialis anterioris muscle to measure intra (IMCL) and extramyocellular lipids (EMCL), quadriceps and handgrip strength, 12 minute walking distance and serum biochemistry (haemoglobin, lipoproteins, creatinine, ultrasensitive C Reactive Protein, fasting and post glucose insulin and glucose concentrations, to assess IS). Physical activity was estimated by actigraphy. Results: 23 men were eutrophic, 26 were overweight and 7 were obese and mostly sedentary, independent of age. Both IMCL and EMCL were higher in overweight/ obese men. Abdominal fat was negatively associated with IS and positively correlated with muscle lipid accretion (both IMCL and EMCL), but not with age. As expected, older individuals had lower muscle mass and strength, but not more adipose tissue nor intramyocellular lipids, yet were more glucose intolerant. Conclusions: central obesity was associated with IMCL and EMCL infiltration and IR. This type of lipid accretion was not related with ageing nor age-related sarcopenia. Older individuals were more glucose intolerant, which was explained by a decrease of insulin secretion more than adiposity-related IR (AU)

Introducción/Objetivos: medir la infiltración grasa en el músculo esquelético, su asociación con resistencia a la insulina (RI) y con masa y función muscular, en hombres chilenos de diferente edad y composición corporal. Nuestra hipótesis era que habría mas acumulación de grasa en el tejido muscular entre las personas de mayor edad y peso, lo cual deterioraría la sensibilidad a la insulina (SI) y afectaría negativamente la masa y la función muscular, ambas características del proceso de envejecimiento. Métodos: se estudiaron hombres sanos (38 < 55 anos y 18 > 65 anos), que fueron sometidos a mediciones antropométricas, evaluación de la composición corporal mediante densitometría radiológica (DEXA), espectroscopia de resonancia nuclear magnética en el músculo tibial anterior para medir lípidos intra (LIM) y extramiocelulares (LEM), fuerza de mano y cuadriceps, test de 12 minutos y bioquímica sérica (glicemia, hemoglobina, lipoproteínas, creatinina y proteína C reactiva ultrasensible en ayunas, ademas de glucosa e insulina post carga de glucosa para evaluar SI). La actividad física se estimo mediante actigrafía. Resultados: 23 hombres eran eutróficos, 26 tenían sobrepeso y 7 eran obesos, todos eran sedentarios según el registro actigráfico, independiente de la edad. Tanto LIM como LEM resultaron mas altos entre los hombres con sobrepeso / obesidad. La grasa abdominal se asocio negativamente con la SI y se correlaciono positivamente con la acumulación de grasa en el músculo (tanto LIM como LEM), pero no con la edad. Como era de esperar, las personas mayores tenían menor masa magra y fuerza, pero no mas tejido adiposo ni lípidos intramiocelulares, aunque eran mas intolerantes a la glucosa. Conclusiones: La obesidad central se asocio con infiltración de grasa intramuscular y con RI. Esta distribución adiposa no se relaciono con edad ni con sarcopenia asociada al envejecimiento. Las personas mayores resultaron mas intolerantes a la glucosa, explicable por una disminución de la secreción de insulina mas que por RI relacionada con mayor adiposidad (AU)

Evaluación de un programa ministerial para manejo del síndrome metabólico en adultos con sobrepeso y obesidad / Evaluation of a program for management of metabolic syndrome in adults with overweight and obesity

Background: The Chilean Ministry of Health developed a healthy lifestyles intervention directed to adults with overweight and cardiovascular risk factors, called "Program on Healthy Eating and Physical Activity" (PASAF). Aim: To evaluate the impact of PASAF on nutritional status and metabolic parameters. Patients and Methods: We analyzed databases from three primary care centers belonging to a municipality of Metropolitan Santiago. We selected adults enrolled in the PASAF during three years (2007-2009). The program lasted four months and included an assessment of anthropometric and metabolic parameters at baseline and at the end, eight workshops with a nutritionist, seven with a psychologist and 32 sessions of physical activity. Result: We evaluated 526 subjects aged ≥ 18 years (93% females), of whom 85.6% attended the last appointment for assessment. Analyzing available data, attendance to workshops was < 50% of the scheduled sessions. Weight, body mass index and waist circumference decreased significantly (median: -1.4 kg, -0.6 kg/m² and -3 cm, respectively). The median weight loss was 1.8% of initial weight and 17.1% of participants experienced a decrease ≥ 5% of their initial weight. There were significant improvements in lipid levels and blood pressure among participants with lower initial excess weight. A reduction in fasting blood glucose was observed only among subjects who lost ≥ 5% of their initial weight. Conclusions: The PASAF modestly reduced nutritional parameters. Correction of metabolic parameters was especially effective in less obese subjects. The attendance to workshops was low.

Variability of formulas to assess insulin sensitivity and their association with the Matsuda index / Variabilidad de las fórmulas que evalúan la sensibilidad a la insulina y su asociación con el Índice Matsuda

Objective: To assess the individual variability of HOMA and QUICKI indexes for the assessment of insulin resistance, using three fasting blood samples obtained within 30 minutes. Research methods & procedures: Data from 80 participants aged 41.5 ± 15 years (26 females), who underwent an oral glucose tolerance test to calculate Matsuda index, were used. Every participant had three fasting blood samples obtained within 30 minutes and four blood samples obtained at 30, 60, 90 and 120 minutes after a 75 g oral glucose load. Insulin and glucose were measured in each sample. HOMA and QUICKI indexes were calculated using the nine possible combinations of the three fasting blood samples. Matsuda index was calculated with all samples obtained. Results: Median values of HOMA-IR, HOMA-β, QUICKI and Matsuda indexes were 1.9, 117.9, 0.35 and 3.71 arbitrary units, respectively. The individual variation coefficients of HOMA-IR, HOMA-β and QUICKI were 11.8 (7.8-18.9), 15 (10.2-22.9) and 1.8 (8.8-21.9) % respectively. When compared with Matsuda index, the R squared values of HOMA-IR, HOMA-β and QUICKI were 0.46, 0.2 and 0.71, respectively. Conclusions: Among fasting indexes for insulin resistance, QUICKI had the lower variation coefficient and the higher correlation with Matsuda index (AU)

Objetivo: Evaluar la variabilidad individual de los índices HOMA y QUICKI para resistencia a insulina, utilizando tres muestras de sangre en ayunas obtenidas en un período de 30 minutos. Material y métodos: Se utilizaron datos provenientes de 80 participantes de 41.5 ± 15 años de edad (26 mujeres) a quienes se les efectuó una prueba de tolerancia a glucosa oral para calcular el índice de Matsuda. A cada participante se le tomaron tres muestras de sangre en ayunas en un período de 30 minutos y cuatro muestras a los 30, 60, 90 y 120 minutos después de una carga oral de 75 g de glucosa. En cada muestra se midieron los niveles de insulina y glucosa. Los índices HOMA y QUICKI se calcularon utilizando las nueve combinaciones posibles con las tres muestras obtenidas en ayunas. El índice de Matsuda se calculó utilizando todas las muestras. Resultados: Las medianas de los índices HOMA-IR, HOMA-β, QUICKI y Matsuda fueron 1,9, 117,9, 0,35 and 3,71 unidades arbitrarias, respectivamente. Los coeficientes de variación individual del HOMA-IR, HOMA-β y QUICKI fueron 11,8 (7,8-18,9), 15 (10,2-22,9) and 1,8 (8,8-21,9) %, respectivamente. Comparados con el índice de Matsuda, los valores de R2 para el HOMA-IR, HOMA-β y QUICKI fueron 0,46, 0,2 y 0,71, respectivamente. Conclusiones: De los índices que utilizan muestras en ayunas para determinar resistencia a insulina, el QUICKI es el que tiene el menor coeficiente de variación y la mejor correlación con el índice de Matsuda (AU)

Variability of glycemic and insulin response to a standard meal, within and between healthy subjects / Variabilidad de las respuestas glucémica y de insulina, intra e intersujeto, a una comida estándar en individuos sanos

Aim: To test the variability within and between subject of glycemic response test following the ingestion of a standard food. Material and methods: Glucose and insulin response of a standard meal (white bread) was performed in ten healthy volunteers and repeated under identical conditions for 6 times. Blood glucose and insulin levels were measured in the fasted state and over the 180 min following commencement of consumption of the foods The Area Under the Curve (AUC) for glucose and insulin was calculated for the values above baseline for the 3hour period following the standard meal. Within and between coefficient of variation was calculated. Results: The total intra-individual variation of the gAUC was 51.8% range 24.9 to 91.4%. The inter-individual variation of the gAUC in the complete study was 75.2% . The total intra-individual variation of the iAUC was 51.9%. ranged: 7.7 to 103%. The inter-individual variation in the complete study was 86%. Conclusion: Glucose and insulin response to a reference food has low reliability, therefore limits its clinical utility for individual dietary prescription (AU)

Objetivo: Evaluar la variabilidad inter e intra individual de la respuesta glicémica frente a la ingestión de un alimento estándar en sujetos sanos. Material y métodos: Se midió en 6 oportunidades la respuesta glicémica e insulínica de un alimento estándar (pan batido blanco) en 10 voluntarios sanos. A cada sujeto se le midió la glicemia e insulina en ayuno a los 15, 45, 60, 90, 120, 150 y 180 minutos, después de ingerir el alimento estándar. Posteriormente se calculó el área bajo la curva (ABC) de glicemia e insulina en cada sujeto. Se calculó el coeficiente de variación intra e inter individual del ABC de glucosa e insulina. Resultados: La variación intra-individual total del ABC de glucosa fue de 51,8% (rango: 24,9%-91,4%). La variación inter-individual del ABC de glucosa total fue 75,2%. La variación intra-individual del ABC de insulina fue de 51,9% (rango: 7,7%-103%). La variación inter-individual del ABC de insulina total fue de 86%. Conclusión: La respuesta glicémica e insulínica a un alimento estándar tiene poca reproducibilidad, tanto intra e inter individual. En consecuencia, su utilidad clínica para la prescripción dietaria individual, está limitada (AU)

Calidad de vida en pacientes operadas de bypass gástrico hace más de un año: Influencia del nivel socioeconómico / Quality of life of patients subjected to gastric bypass more than one year ago: Influence of socioeconomic status

Background: The study of the effects of bariatric surgery on quality of life in patients of different socioeconomic levels (SEL) is worthwhile. Aim: To study quality of life (QoL), eating behavior, depressive symptoms and sexuality in patients subjected to a gastric bypass (GBP) more than 1 year before. Material and methods: The sample was composed of 33 GPB patients (19 high SEL and 14 low SEL), and 27 non-operated women (18 high SEL and 9 low SEL) of similar weight and age, as controls. Assessment included medical history anthropometry radiological densitometry. Eating behavior was assessed using the three factor eating questionnaire, quality of life using SF-36 and the Bariatric Analysis of Reporting Outcome System (BAROS) depressive symptoms were assessed using the Beck scale version II and sexual behavior using the female sexual function index (FSFI). Results: QoL was lower in operated patients from low SEL, especially when compared to high SEL control women. Operated patients had a predominantly restrictive pattern of eating behavior. Eating behavior disorders were detected in 5 of 33 operated patients versus 4 of 27 controls (p =ns). Sexual function was absent or dysfunctional in 22 operated versus 8 controls (p =0,02). No significant differences were observed for depressive symptoms, between operated patients and controls. Conclusions: In the long term, QoL of bariatric patients, especially those from low SEL, is inferior to control women and Chilean general population. Operated patients have restrictive eating patterns and lower sexual satisfaction indexes. Frequency of depressive symptoms was high both in bariatric and control women.

Evaluación a largo plazo del estado nutricional, composición corporal y densidad mineral ósea en mujeres operadas de bypass gástrico: impacto del nivel socioeconómico / Nutritional status, body composition and bone mineral density in gastric bypass females: impact of socioeconomic level

Background: Roux-en-Y gastric bypass (RYGBP) has had a positive impact on co-morbidities associated with obesity. However, in the long-term it can induce micronutrient deficiencies. Aim: To perform a complete nutritional assessment in a group of women previously operated of RYGBP, from different socioeconomic levéis (SEL). Patients and Methods: Thirty three women (19 high SEL and 14 low SEL), were assessed by dietary recalls, anthropometric measurements, muscle strength, bone mineral density, routine clinical laboratory, serum levéis of vitamin B12, 250H-vitamin D, Mate, calcium, ferritine, ceruloplasmin and indicators ofbone turnover (parathohormone, osteocalcin and urinary pyridinolines). Their valúes were compared to those of 30 control women (18 high SEL and 12 low SEL). Results: Low SEL operated women consumed fewer vitamin and mineral supplements compared with their high SEL pairs. No cases of vitamin B12, folie acid or copper deficiencies were detected. Frequency of iron deficieney was similar in patients and controls. Vitamin D insufficieney was higher amongpatients than in controls (p =0,04 7), regardless SEL. Patients had also a higher frequency of high serum PTH and osteocalcin and urinary pyridinoline levéis. However, no differences in bone mineral density were observed between operated women and controls. Conclusions: Vitamin and mineral deficiencies were lower than expected among operated women. However, problems associated with vitamin D deficieney were highly prevalent among patients operated of RYGBP, irrespective SEL. These alterations were only detectable through speciñe markers at this stage, because they did not transíate into lower bone mineral density (BMD) of surgicalpatients, probably due to the higher pre-operative BMD of these morbidobese patients.

Efecto del probiótico Saccharomyces boulardii en la prevención de la diarrea asociada con antibióticos en adultos ambulatorios en tratamiento con amoxicilina / Effect of probiotic Saccharomyces boulardii on prevention of antibiotic-associated diarrhea in adult outpatients with amoxicillin treatment

Background: Antibiotic-associated diarrhea is one of the most common adverse effects of antimicrobials. Any antimicrobial can potentially produce diarrhea but beta-lactamics have a higher risk. Among these, amoxicillin is widely indicated in ambulatory practice. One ofthe alternatives suggested to prevent antibiotic-associated diarrhea, is the use of the probiotic Saccharomyces boulardii. Aim: To evalúate whether the concomitant use of Saccharomyces boulardii and amoxicillin can prevent antibiotic associated diarrhea in ambulatory adults with acute infections diseases, without provoking other adverse effects. Material and Methods: Eighty six adults (aged 15 to 81 years) with acute infectious diseases, excluding those arising in the gastrointestinal tract, that received a prescription of oral amoxicillin for 5 to 10 days, were included. In a controlled randomized, double blind trial, 41 patients were assigned to receive lyophilized Saccharomyces boulardii (500 mg/day) duríng 12 days, and 45 patients were assigned to placebo for the same period. Results: Ten percent of patients (9/86) reported acute diarrhea, 9,8 percent (4/41) in the experimental group and 11.196 (5/45) in the control group (p = 100). No adverse effects were associated to the use of the probiotic. Conclusions: Saccharomyces boulardii (500 mg/day) did not prevent diarrhea related to amoxicillin.

Determinación de los índices glicémicos y de insulina en fórmulas para alimentación enteral en adultos sanos / Glycemic and insulin indices of tube feeding formulas in healthy adults

Background: In acute illnesses, plasma glucose levels are often increased and generally parallel the severity of stress. Hyperglycemia caused by reduced insulin sensitivity and reduced insulin secretion is associated with increased susceptibility to infections. Maintaining blood glucose levels at or below 110 mg/dl reduces morbidity and mortality in critically ill patients. Aim: To measure the glucose and insulin responses of four commercially available enteral formulas compared with a standard meal reference product. Material and Methods: The glycemic index (GI) and the insulin index (II) were determined in a randomized, cross over protocol in 38 healthy volunteers between 18 and 46 years of age. Each subject underwent five tests: three with the standard meal (bread) and two with the study products. The enteral formulas were Clinutren HPR (whole protein of high protein value), Crucial® (casein peptide based formula), Peptamen®, (whey peptide based formula), Glytrol® (formula for diabetics with whole protein with fiber). Each study product was evaluated 10 times. Results: The diabetic formula and the high protein energy dense formulas induced a significantly lower GI (p <0.02) compared with the standard meal. The GI response did not appear to be due to enhanced insulin secretion. The other tested formulas had lower GI than the standard meal, but in addition they exhibited increased II The whey based peptide formulation produced the highest insulin response (p <0.03). Conclusions: Both GI and II are related to the concentration, form and type of protein contained in the enteral formula. The whey peptide formulation produced a low GI with the highest insulin index. Based on the low GI of these enteral products, all can be useful to provide nutritional support during metabolic stress, without adding an additional challenge to blood glucose management.

Rabbit IgG antibodies against Phospholipase A2 from Crotalus durissus terrificus neutralize the lethal activity of the venom

Crotalus durissus terrificus (C.d.t.) (South American rattlesnake) venom possesses myotoxic and neurotoxic activities, both of which are also expressed by crotoxin, the principal toxin of this venom. Crotoxin contains a basic phospholipase A2 (PLA2) and a non toxic acidic protein, crotapotin. We have produced and investigated the ability of IgG antibodies raised in rabbits against PLA2 to neutralize the lethality of the whole venom. PLA2 was isolated by gel filtration chromatography (Sephadex G-75). Specific antibodies were obtained by subcutaneous and intramuscular inoculation of PLA2 (700 µg) with Freund adjuvant. Groups of six mice (20 + 2 g) were inoculated with 0.5 ml i.p. of C. d. t. venom (4 µg) or a mixture of venom that had been preincubated with the desired volume of IgG antibodies. Mortality, recorded 24 and 48 h after inoculation, showed that IgG anti-PLA2 were more effective than anticrotalic serum in neutralizing the lethal activity. These results demonstrate that it could be possible to obtain an anti-venom made by specific antibodies with a high level of protection against the lethal component of C.d.t. venom, and/or the inclusion of these antibodies as a supplement in heterologous anti-venoms

El veneno de Crotalus durissus terrificus (C.d.t.) (Cascabel de Sud América) posee actividad miotóxica y neurotóxica, actividades que también exhibe el complejo crotoxina, principal componente tóxico de este veneno. El complejo crotoxina está constituido por una fosfolipasa A2 básica (PLA2) y una proteína acídica no tóxica, el crotapotín. En este trabajo se estudió la capacidad neutralizante de anticuerpos IgG anti-PLA2 sobre la letalidad inducida por el veneno entero. El antígeno PLA2, fue aislado por cromatografía de filtración en gel (Sephadex G-75). Se inocularon conejos machos por vía subcutánea e intramuscular, con 700 µg de PLA2 y adyuvante para la obtención de anticuerpos específicos. La capacidad neutralizante del antisuero se analizó en ratones por inoculación con diluciones de veneno entero preincubado con un volumen adecuado de anticuerpos IgG anti-PLA2. Se inocularon ratones controles con 0.5 ml i.p. de veneno (4 µg.ml-1). El número de muertes fue contabilizado a las 24 y 48 h posteriores a la inoculación, demostrándose que la capacidad neutralizante de los anticuerpos IgG anti-PLA2 fue superior a la obtenida con el antiveneno crotálico. Los resultados obtenidos demuestran la potencial aplicación de antivenenos constituidos por anticuerpos específicos contra PLA2, y/o la inclusión de estos anticuerpos como suplementos en antivenenos polivalentes

Rabbit IgG antibodies against Phospholipase A2 from Crotalus durissus terrificus neutralize the lethal activity of the venom

Crotalus durissus terrificus (C.d.t.) (South American rattlesnake) venom possesses myotoxic and neurotoxic activities, both of which are also expressed by crotoxin, the principal toxin of this venom. Crotoxin contains a basic phospholipase A2 (PLA2) and a non toxic acidic protein, crotapotin. We have produced and investigated the ability of IgG antibodies raised in rabbits against PLA2 to neutralize the lethality of the whole venom. PLA2 was isolated by gel filtration chromatography (Sephadex G-75). Specific antibodies were obtained by subcutaneous and intramuscular inoculation of PLA2 (700 Ag) with Freund adjuvant. Groups of six mice (20 + 2 g) were inoculated with 0.5 ml i.p. of C. d. t. venom (4 Ag) or a mixture of venom that had been preincubated with the desired volume of IgG antibodies. Mortality, recorded 24 and 48 h after inoculation, showed that IgG anti-PLA2 were more effective than anticrotalic serum in neutralizing the lethal activity. These results demonstrate that it could be possible to obtain an anti-venom made by specific antibodies with a high level of protection against the lethal component of C.d.t. venom, and/or the inclusion of these antibodies as a supplement in heterologous anti-venoms (AU)

El veneno de Crotalus durissus terrificus (C.d.t.) (Cascabel de Sud América) posee actividad miotóxica y neurotóxica, actividades que también exhibe el complejo crotoxina, principal componente tóxico de este veneno. El complejo crotoxina está constituido por una fosfolipasa A2 básica (PLA2) y una proteína acídica no tóxica, el crotapotín. En este trabajo se estudió la capacidad neutralizante de anticuerpos IgG anti-PLA2 sobre la letalidad inducida por el veneno entero. El antígeno PLA2, fue aislado por cromatografía de filtración en gel (Sephadex G-75). Se inocularon conejos machos por vía subcutánea e intramuscular, con 700 Ag de PLA2 y adyuvante para la obtención de anticuerpos específicos. La capacidad neutralizante del antisuero se analizó en ratones por inoculación con diluciones de veneno entero preincubado con un volumen adecuado de anticuerpos IgG anti-PLA2. Se inocularon ratones controles con 0.5 ml i.p. de veneno (4 Ag.ml-1). El número de muertes fue contabilizado a las 24 y 48 h posteriores a la inoculación, demostrándose que la capacidad neutralizante de los anticuerpos IgG anti-PLA2 fue superior a la obtenida con el antiveneno crotálico. Los resultados obtenidos demuestran la potencial aplicación de antivenenos constituidos por anticuerpos específicos contra PLA2, y/o la inclusión de estos anticuerpos como suplementos en antivenenos polivalentes (AU)

Rabbit IgG antibodies against Phospholipase A2 from Crotalus durissus terrificus neutralize the lethal activity of the venom

Crotalus durissus terrificus (C.d.t.) (South American rattlesnake) venom possesses myotoxic and neurotoxic activities, both of which are also expressed by crotoxin, the principal toxin of this venom. Crotoxin contains a basic phospholipase A2 (PLA2) and a non toxic acidic protein, crotapotin. We have produced and investigated the ability of IgG antibodies raised in rabbits against PLA2 to neutralize the lethality of the whole venom. PLA2 was isolated by gel filtration chromatography (Sephadex G-75). Specific antibodies were obtained by subcutaneous and intramuscular inoculation of PLA2 (700 Ag) with Freund adjuvant. Groups of six mice (20 + 2 g) were inoculated with 0.5 ml i.p. of C. d. t. venom (4 Ag) or a mixture of venom that had been preincubated with the desired volume of IgG antibodies. Mortality, recorded 24 and 48 h after inoculation, showed that IgG anti-PLA2 were more effective than anticrotalic serum in neutralizing the lethal activity. These results demonstrate that it could be possible to obtain an anti-venom made by specific antibodies with a high level of protection against the lethal component of C.d.t. venom, and/or the inclusion of these antibodies as a supplement in heterologous anti-venoms (AU)

El veneno de Crotalus durissus terrificus (C.d.t.) (Cascabel de Sud América) posee actividad miotóxica y neurotóxica, actividades que también exhibe el complejo crotoxina, principal componente tóxico de este veneno. El complejo crotoxina está constituido por una fosfolipasa A2 básica (PLA2) y una proteína acídica no tóxica, el crotapotín. En este trabajo se estudió la capacidad neutralizante de anticuerpos IgG anti-PLA2 sobre la letalidad inducida por el veneno entero. El antígeno PLA2, fue aislado por cromatografía de filtración en gel (Sephadex G-75). Se inocularon conejos machos por vía subcutánea e intramuscular, con 700 Ag de PLA2 y adyuvante para la obtención de anticuerpos específicos. La capacidad neutralizante del antisuero se analizó en ratones por inoculación con diluciones de veneno entero preincubado con un volumen adecuado de anticuerpos IgG anti-PLA2. Se inocularon ratones controles con 0.5 ml i.p. de veneno (4 Ag.ml-1). El número de muertes fue contabilizado a las 24 y 48 h posteriores a la inoculación, demostrándose que la capacidad neutralizante de los anticuerpos IgG anti-PLA2 fue superior a la obtenida con el antiveneno crotálico. Los resultados obtenidos demuestran la potencial aplicación de antivenenos constituidos por anticuerpos específicos contra PLA2, y/o la inclusión de estos anticuerpos como suplementos en antivenenos polivalentes (AU)

Mice plasma fibrinogen consumption by thrombin-like enzyme present in rattlesnake venom from the north-east region of Argentina.

Due to variability of venom components from the same species of snakes that inhabit different regions, particular properties of the venom of Crotalus durissus terrificus that inhabits the North-East of Argentina were studied. Gyroxin, a thrombin-like enzyme, was isolated from this venom by gel filtration and affinity chromatography, it was found to be homogeneous according to SDS-PAGE, with a molecular weight of 33 kDa. [quot ]Gyroxin syndrome[quot ] in mice was tested and it showed changes in the animal behavior, confirming that the isolated thrombin-like enzyme is gyroxin. Effects of this enzyme and the crude venom on mice plasmatic fibrinogen levels were determined. The mice plasma fibrinogen decreased rapidly until incoagulability during the first hour after thrombin-like enzyme injection, then reaching its normal level 10 hours after injection; whereas crude venom resulted in a 60

decrease of the mice plasma fibrinogen, reaching its normal level after the same period of time. After 1 hour of gyroxin inoculation, intravascular coagulation was observed in histological cuttings of lung, cardiac muscle and liver. The isolated enzyme showed strong hydrolyzing activity on fibrinogen and fibrin in vitro, whereas the crude venom exhibited weak hydrolyzing activity on both substrates. It is probable that this very low activity is due to the low percentage of the enzyme in the crude venom. Decreasing of plasmatic fibrinogen levels may be due to either the coagulant or hydrolyzing actions of the enzyme.