Hemodiálise veterinária: considerações do colégio brasileiro de nefrologia e urologia veterinárias / Veterinary hemodialysis: considerations of the brazilian college of veterinary nephrology and urology / Hemodiálisis veterinaria: consideraciones del colegio brasileño de nefrología y urología veterinaria

Sendo parte integrante do arsenal de opções terapêuticas na nefrologia veterinária, a hemodiálise aplicada aos animais se popularizou e avançou consideravelmente nos últimos anos, não só no mundo como no Brasil. Neste contexto, visando apresentar as características principais da hemodiálise,o Colégio Brasileiro de Nefrologia e Urologia Veterinárias (CBNUV), em nome da sua subcomissão de Técnicas Dialíticas, traz, neste documento, as informações necessárias para embasar os conhecimentos sobre o tema.(AU)

Being an integral part of the arsenal of therapeutic options in veterinary nephrology, hemodialysis applied to animals has become popular and has advanced considerably in recent years, not only in the world but in Brazil. In this context, intending to present the main characteristics of hemodialysis, the Colégio Brasileiro de Nephrologiae Urologia Veterinárias (CBNUV), on behalf of its subcommittee on Dialytic Techniques, brings, in this document, the necessary information to base knowledge on the subject.(AU)

Siendo parte integral del arsenal de opciones terapéuticas en nefrología veterinaria, la hemodiálisis aplicada a animales se popularizó y avanzó considerablemente en los últimos años, no solo en el mundo sino también en Brasil. En ese contexto, con el fin de presentar las principales características de la hemodiálisis, el Colégio Brasileiro de Nephrologia e Urologia Veterinárias (CBNUV), en representación de su subcomité de Técnicas Dialíticas, trae, en este documento, las informaciones necesarias para fundamentar el conocimiento sobre el tema.(AU)

Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum

Starch processing industries use amylases, accounting for approximately 30% of the world's enzyme market. Previously, an amylase-producing strain of Epicoccum nigrumwas isolated from maize grains. Although E. nigrumamylase production is already reported in the literature, no published data on production optimization or characterization of the produced enzyme exists. The objectives of this work were to improve the amylase production by theE. nigrumPG 16 strain and to purify and characterize the produced enzyme. The E. nigrumPG 16 amylase production best conditions in submerged culture were: inoculum of 4% (v v-1) of a five-days-old stationary culture homogenate, agitation at 100 rpm, 25°C, natural light, 72 hours of incubation, starch as thecarbon source, and an initial medium pH of 7.0. A molecular exclusion chromatography profile has shown the production of only one amylase, which was partially purified with ammonium precipitation and dialysis. The enzyme optima pH and temperature are 6.0 and 50°C, respectively. The partially purified enzyme lostits activity when incubated for 30 min in temperatures above 40°C, presenting a T50of 46.25°C. The KMand Vmaxof the partially purified enzyme are 1.72 mg mL-1of starch and 0.15 mgmin-1of degraded starch, respectively. The ion Ca2+slightly activated the studied enzyme. The ions Cu2+, Zn2+, and Fe3+and the detergents SDS and Tween 80 acted as inhibitors of the studied enzyme. The partially purified enzyme released glucose from p-nitrophenyl α-D-glucopyranoside (p-NPG). Glucose was the enzyme's main product from starch hydrolysis, as evidenced by thin-layer chromatography. The E. nigrumPG 16 studied enzyme is a glucoamylase and represents an alternative for enzymatic starch hydrolysis.(AU)

Qualidade da água utilizada em serviços de hemodiálise antes e após passar por sistema de tratamento em Santarém, Oeste do Pará, Amazônia / Quality of the water used in hemodialysis services before and after going through a treatment system in Santarém, Western Pará, Amazonia

A água utilizada em serviços de hemodiálise passa por um sistema de tratamento a fim de garantir que os parâmetros microbiológicos e físico-químicos estejam adequados e proporcionem segurança ao paciente. Este trabalho avaliou a qualidade da água antes e após esta passar por sistema de tratamento em dois serviços (A e B) de hemodiálise em Santarém. Foram coletadas amostras de água em três pontos: Ponto 1 (P1) após saída do poço, Ponto 2 (P2) na saída do tratamento e Ponto 3 (P3) na máquina de hemodiálise. Testes microbiológicos e físico-químicos foram realizados e os resultados mostraram que não houve crescimento de coliformes totais e Escherichia coli nas amostras. No serviço A, houve contaminação por bactérias heterotróficas em 87% das amostras, destas, 27% ocorreram no P3. O pH mostrou-se menor que 6,0 no P1 nos dois serviços, normalizando os seus valores no P3. No serviço A, concentrações superiores ao recomendado de alumínio e nitrato foram verificadas em 100% das amostras coletadas no P1, os valores foram normalizados na saída do tratamento (P2). A pesquisa demonstrou que, para os parâmetros estudados, a qualidade da água, após passar por sistema de tratamento, é satisfatória, com exceção das bactérias heterotróficas no serviço A. (AU)

The water used in hemodialysis services goes through a treatment system in order to ensure that the microbiological and physicochemical parameters are adequate and provide safety to the patient. This work evaluated the quality of water before and after it went through a treatment system in two hemodialysis services (A and B) in Santarém. Water samples were collected at three points: Point 1 (P1) after leaving the well, Point 2 (P2) leaving the treatment and Point 3 (P3) on the hemodialysis machine. Microbiological and physical-chemical tests were performed and the results showed no growth of total coliforms and Escherichia coli in the samples. In service A, there was contamination by heterotrophic bacteria in 87% of the samples, 27% of which occurred in P3. The pH was less than 6.0 in P1 in both services, normalizing its values in P3. Service A, concentrations higher than the recommended amount of aluminum and nitrate were verified in 100% of the samples collected in P1, the values were normalized at the end of the treatment (P2). The research demonstrated that, for the studied parameters, the water quality, after going through a treatment system, is satisfactory, with the exception of heterotrophic bacteria in service A. (AU)

Chronic kidney failure following lancehead bite envenoming: a clinical report from the Amazon region

Snakebite envenoming can be a life-threatening condition, for which emergency care is essential. The Bothrops (lancehead) genus is responsible for most snakebite-related deaths and permanent loss of function in human victims in Latin America. Bothrops spp. venom is a complex mixture of different proteins that are known to cause local necrosis, coagulopathy, and acute kidney injury. However, the long-term effects of these viper envenomings have remained largely understudied. Case presentation: Here, we present a case report of a 46-years old female patient from Las Claritas, Venezuela, who was envenomed by a snake from the Bothrops genus. The patient was followed for a 10-year period, during which she presented oliguric renal failure, culminating in kidney failure 60 months after the envenoming. Conclusion: In Latin America, especially in Brazil, where there is a high prevalence of Bothrops envenoming, it may be relevant to establish long-term outpatient programs. This would reduce late adverse events, such as chronic kidney disease, and optimize public financial resources by avoiding hemodialysis and consequently kidney transplantation.(AU)

Analysis of hemodialysis water quality from 2008 to 2016 in Minas Gerais, Brazil / Análise da qualidade da água de hemodiálise durante o período de 2008 a 2016 em Minas Gerais, Brasil

Water quality requirements for hemodialysis are crucial for preventing health risks for renal patients. The objective of this study was to assess the quality of the water used for hemodialysis treatments in the state of Minas Gerais. The analytical results for 25 parameters in 1056 hemodialysis water samples from 89 renal dialysis units were analyzed. Data were obtained through the Minas Gerais Hemodialysis Unit Quality Monitoring Programme between July 2008 and December 2016. The effects of the Programme on patient health protection were assessed by evaluating compliance with legal limits and the efficacy of implementing inspections from 2012. Considering the 1056 samples analyzed, 264 (25%) presented at least one parameter in disagreement with current legislation. Conductivity (9.8%), endotoxin (6.9%), fluoride (4.2%) and heterotrophic bacteria (2.2%) were the most critical parameters. Nevertheless, the concentrations of all these parameters (except fluoride) decreased after implementing inspections. The average levels of 15 parameters decreased throughout the study period. The annual percentage of non-compliance decreased from 44% in 2008 to 23% in 2016. It was verified the relevance of the implementation of the Program in the prevention of possible disorders to the patients’ health, indicating the importance of its continuity.

Os requisitos de qualidade da água para a hemodiálise são cruciais para prevenir riscos à saúde de pacientes renais. O objetivo foi avaliar a qualidade da água utilizada nos tratamentos de hemodiálise em Minas Gerais. Os dados foram obtidos por meio do Programa de Monitoramento da Qualidade dos Serviços de Hemodiálise do Estado entre julho de 2008 e dezembro de 2016. Os resultados analíticos para 25 parâmetros em 1056 amostras de água em hemodiálise de 89 unidades renais foram analisados. Os efeitos do Programa na saúde dos pacientes foram avaliados através da conformidade dos limites legais e da eficácia da implementação de inspeções a partir de 2012. Considerando-se as 1056 amostras analisadas, 264 (25%) apresentaram pelo menos um parâmetro em desacordo com a legislação vigente. Condutividade, endotoxina, fluoreto e bactérias heterotróficas foram os parâmetros mais críticos. No entanto, as concentrações de todos esses parâmetros (exceto flúor) diminuíram após a implementação das inspeções. Os níveis médios de 15 parâmetros diminuíram ao longo do período do estudo. O percentual anual de não conformidade diminuiu de 44% (2008) para 23% (2016). Verificou-se a relevância da implantação do Programa na prevenção de possíveis transtornos à saúde dos pacientes, indicando a importância de sua continuidade.

Analysis of hemodialysis water quality from 2008 to 2016 in Minas Gerais, Brazil / Análise da qualidade da água de hemodiálise durante o período de 2008 a 2016 em Minas Gerais, Brasil

Water quality requirements for hemodialysis are crucial for preventing health risks for renal patients. The objective of this study was to assess the quality of the water used for hemodialysis treatments in the state of Minas Gerais. The analytical results for 25 parameters in 1056 hemodialysis water samples from 89 renal dialysis units were analyzed. Data were obtained through the Minas Gerais Hemodialysis Unit Quality Monitoring Programme between July 2008 and December 2016. The effects of the Programme on patient health protection were assessed by evaluating compliance with legal limits and the efficacy of implementing inspections from 2012. Considering the 1056 samples analyzed, 264 (25%) presented at least one parameter in disagreement with current legislation. Conductivity (9.8%), endotoxin (6.9%), fluoride (4.2%) and heterotrophic bacteria (2.2%) were the most critical parameters. Nevertheless, the concentrations of all these parameters (except fluoride) decreased after implementing inspections. The average levels of 15 parameters decreased throughout the study period. The annual percentage of non-compliance decreased from 44% in 2008 to 23% in 2016. It was verified the relevance of the implementation of the Program in the prevention of possible disorders to the patients health, indicating the importance of its continuity.(AU)

Os requisitos de qualidade da água para a hemodiálise são cruciais para prevenir riscos à saúde de pacientes renais. O objetivo foi avaliar a qualidade da água utilizada nos tratamentos de hemodiálise em Minas Gerais. Os dados foram obtidos por meio do Programa de Monitoramento da Qualidade dos Serviços de Hemodiálise do Estado entre julho de 2008 e dezembro de 2016. Os resultados analíticos para 25 parâmetros em 1056 amostras de água em hemodiálise de 89 unidades renais foram analisados. Os efeitos do Programa na saúde dos pacientes foram avaliados através da conformidade dos limites legais e da eficácia da implementação de inspeções a partir de 2012. Considerando-se as 1056 amostras analisadas, 264 (25%) apresentaram pelo menos um parâmetro em desacordo com a legislação vigente. Condutividade, endotoxina, fluoreto e bactérias heterotróficas foram os parâmetros mais críticos. No entanto, as concentrações de todos esses parâmetros (exceto flúor) diminuíram após a implementação das inspeções. Os níveis médios de 15 parâmetros diminuíram ao longo do período do estudo. O percentual anual de não conformidade diminuiu de 44% (2008) para 23% (2016). Verificou-se a relevância da implantação do Programa na prevenção de possíveis transtornos à saúde dos pacientes, indicando a importância de sua continuidade.(AU)

Intermittent hemodialysis, an alternative treatment for ovine with acute-on-chronic kidney disease / Hemodiálise intermitente, um tratamento alternativo para ovinos em doença renal crônica agudizada / Hemodialisis intermitente, un tratamiento alternativo para ovinoscon enfermedad renal aguda en crónica

Chronic Kidney Disease (CKD) is a syndrome composed of numerous comorbidities such as the failure of excretion of toxins and nitrogenous compounds from metabolism, as well as the failure to maintain endocrine, water, electrolyte and acid-base homeostasis in all mammals. As it is a non-regenerative and progressive disease, any endogenous or exogenous insults in the organ in question will lead to the intensification of all these mentioned disorders associated with decreased urinary output. Several studies in humans and animals recommend the use of renal replacement therapy, especially in those who already have CKD, and who are in uremic syndrome. This report aims to demonstrate that the use of intermittent hemodialysis (HI) in sheep is a therapeutic alternative in those in renal failure. A six-year-old Bergamasca sheep was presented with lateral decubitus, dyspnea, dark liquid diarrhea, apathy, anorexia and pulmonary crepitation on auscultation. Laboratory tests showed azotemia and electrolyte changes such as hypocalcemia, hypokalemia, hypochloremia, isosthenuria (1,012), pyuria, glycosuria, and a protein: creatinine ratio of 0.88. In ultrasonography, the real resistivity index was above 0.54, being suggestive of loss of renal perfusion. The treatment chosen consisted of electrolytic and fluid replacement, associated with HI. After HI, the sheep showed great clinical and laboratory improvement as expected, however, despite not having survived, the histopathological findings of the necropsy pointed out segments of glomerulosclerosis, thus reinforcing the hypothesis that the animal already had a CKD which was aggravated due to the association of hypovolemia, hypotension by anesthetic drugs and the indiscriminate use of NSAIDs. Despite the death of the sheep, HI again proved to be a therapy of rapid clinical...

A Doença Renal Crônica (DRC) é uma síndrome composta por inúmeras comorbidades como a falha da excreção de toxinas e compostos nitrogenados oriundos do metabolismo, bem como da falha na manutenção da homeostase endócrina, hídrica, eletrolítica e ácido-básico de todos os mamíferos. Por ser uma doença não-regenerativa e progressiva, quaisquer insultos endógenos ou exógenos no órgão em questão propiciará a intensificação de todas estas desordens mencionadas associadas a diminuição do débito urinário. Diversos estudos em humanos e animais recomendam o emprego de terapia de substituição renal, especialmente naqueles que já apresentam a DRC, e que se encontram em síndrome urêmica. Este relato tem como objetivo demonstrar que o emprego da hemodiálise intermitente (HI) em ovinos é uma alternativa terapêutica naqueles em falência renal. Uma ovelha Bergamácia de seis anos foi encaminhada em decúbito lateral, dispneia, diarreia líquida escura, apatia, anorexia e crepitação pulmonar à auscultação. Exames laboratoriais demonstraram azotemia e alterações eletrolíticas como hipocalcemia, hipocalemia, hipocloremia, isostenúria (1.012), piúria, glicosúria, e razão proteína: creatinina de 0,88. Em ultrassonografia, o index de resistividade real, apresentou-se acima de 0,54, sendo sugestivo de perda de perfusão renal. O tratamento eleito constituiu na reposição eletrolítica e volêmica, associada a HI. Após a HI, a ovelha apresentou grande melhora clínica e laboratorial como esperado, no entanto, apesar de não ter sobrevivido, os achados histopatológicos da necropsia apontaram segmentos de glomeruloesclerose, reforçando assim a hipótese de que o animal já apresentava uma DRC a qual agudizou devido a associação da hipovolemia, hipotensão por fármacos anestésicos e ao uso indiscriminado de AINES. Apesar do óbito da ovelha, a HI demonstrou novamente ser uma terapia de rápida melhora clínica e laboratorial do paciente comparada ao tratamento clínico convencional, assim sendo uma importante alternativa...

La enfermedad renal crónica (ERC) es un síndrome compuesto por numerosas comorbilidades, como el fracaso de la excreción de toxinas y compuestos nitrogenados del metabolismo, así como el fracaso para mantener la homeostasis endocrina, hidrica, electroliticay ácido-base en todos los mamíferos. Como es una enfermedad no regenerativa y progresiva, cualquier insulto endógeno o exógeno en el órgano en cuestión conducirá a la intensificación de todos estos trastornos mencionados asociados con la disminución del gasto urinario. Varios estudios en humanos y animales recomiendan el uso de la terapia de reemplazo renal, especialmente en aquellos que ya tienen ERC y que padecen síndrome urémico. Este informe tiene como objetivo demostrar que el uso de hemodiálisis intermitente (HI) en ovejas es una alternativa terapéutica en aquellos con insuficiencia renal. Una oveja Bergamasca de seis años fue apresentadaen decúbito lateral, disnea, diarrea líquida oscura, apatía, anorexia y crepitación pulmonar en la auscultación. Las pruebas de laboratorio mostraron alteraciones de azotemia y electrolitos como hipocalcemia, hipocalemia, hipocloremia, isostenuria (1,012), piuria, glucosuria y una relación proteína: creatinina de 0.88. En la ecografía, el índice de resistividad real fue superior a 0,54, lo que sugiere una pérdida de perfusión renal. El tratamiento elegido consistió en reemplazo electrolítico y de fluidos, asociado con HI. Después de HI,la oveja mostró una gran mejoría clínica y de laboratorio como se esperaba, sin embargo, a pesar de no haber sobrevivido, los hallazgos histopatológicos de la necropsia señalaron segmentos de glomeruloesclerosis, lo que refuerza la hipótesis de que el animal ya tenía una ERC que se agravó debido a la asociación de hipovolemia, hipotensión por fármacos anestésicos y el uso indiscriminado de AINE. A pesar de la muerte de la oveja, HI nuevamente demostró ser una terapia de rápida mejora clínica y de laboratorio del paciente en comparación con el tratamiento clínico convencional...

Intermittent hemodialysis, an alternative treatment for ovine with acute-on-chronic kidney disease / Hemodiálise intermitente, um tratamento alternativo para ovinos em doença renal crônica agudizada / Hemodialisis intermitente, un tratamiento alternativo para ovinoscon enfermedad renal aguda en crónica

Chronic Kidney Disease (CKD) is a syndrome composed of numerous comorbidities such as the failure of excretion of toxins and nitrogenous compounds from metabolism, as well as the failure to maintain endocrine, water, electrolyte and acid-base homeostasis in all mammals. As it is a non-regenerative and progressive disease, any endogenous or exogenous insults in the organ in question will lead to the intensification of all these mentioned disorders associated with decreased urinary output. Several studies in humans and animals recommend the use of renal replacement therapy, especially in those who already have CKD, and who are in uremic syndrome. This report aims to demonstrate that the use of intermittent hemodialysis (HI) in sheep is a therapeutic alternative in those in renal failure. A six-year-old Bergamasca sheep was presented with lateral decubitus, dyspnea, dark liquid diarrhea, apathy, anorexia and pulmonary crepitation on auscultation. Laboratory tests showed azotemia and electrolyte changes such as hypocalcemia, hypokalemia, hypochloremia, isosthenuria (1,012), pyuria, glycosuria, and a protein: creatinine ratio of 0.88. In ultrasonography, the real resistivity index was above 0.54, being suggestive of loss of renal perfusion. The treatment chosen consisted of electrolytic and fluid replacement, associated with HI. After HI, the sheep showed great clinical and laboratory improvement as expected, however, despite not having survived, the histopathological findings of the necropsy pointed out segments of glomerulosclerosis, thus reinforcing the hypothesis that the animal already had a CKD which was aggravated due to the association of hypovolemia, hypotension by anesthetic drugs and the indiscriminate use of NSAIDs. Despite the death of the sheep, HI again proved to be a therapy of rapid clinical...(AU)

A Doença Renal Crônica (DRC) é uma síndrome composta por inúmeras comorbidades como a falha da excreção de toxinas e compostos nitrogenados oriundos do metabolismo, bem como da falha na manutenção da homeostase endócrina, hídrica, eletrolítica e ácido-básico de todos os mamíferos. Por ser uma doença não-regenerativa e progressiva, quaisquer insultos endógenos ou exógenos no órgão em questão propiciará a intensificação de todas estas desordens mencionadas associadas a diminuição do débito urinário. Diversos estudos em humanos e animais recomendam o emprego de terapia de substituição renal, especialmente naqueles que já apresentam a DRC, e que se encontram em síndrome urêmica. Este relato tem como objetivo demonstrar que o emprego da hemodiálise intermitente (HI) em ovinos é uma alternativa terapêutica naqueles em falência renal. Uma ovelha Bergamácia de seis anos foi encaminhada em decúbito lateral, dispneia, diarreia líquida escura, apatia, anorexia e crepitação pulmonar à auscultação. Exames laboratoriais demonstraram azotemia e alterações eletrolíticas como hipocalcemia, hipocalemia, hipocloremia, isostenúria (1.012), piúria, glicosúria, e razão proteína: creatinina de 0,88. Em ultrassonografia, o index de resistividade real, apresentou-se acima de 0,54, sendo sugestivo de perda de perfusão renal. O tratamento eleito constituiu na reposição eletrolítica e volêmica, associada a HI. Após a HI, a ovelha apresentou grande melhora clínica e laboratorial como esperado, no entanto, apesar de não ter sobrevivido, os achados histopatológicos da necropsia apontaram segmentos de glomeruloesclerose, reforçando assim a hipótese de que o animal já apresentava uma DRC a qual agudizou devido a associação da hipovolemia, hipotensão por fármacos anestésicos e ao uso indiscriminado de AINES. Apesar do óbito da ovelha, a HI demonstrou novamente ser uma terapia de rápida melhora clínica e laboratorial do paciente comparada ao tratamento clínico convencional, assim sendo uma importante alternativa...(AU)

La enfermedad renal crónica (ERC) es un síndrome compuesto por numerosas comorbilidades, como el fracaso de la excreción de toxinas y compuestos nitrogenados del metabolismo, así como el fracaso para mantener la homeostasis endocrina, hidrica, electroliticay ácido-base en todos los mamíferos. Como es una enfermedad no regenerativa y progresiva, cualquier insulto endógeno o exógeno en el órgano en cuestión conducirá a la intensificación de todos estos trastornos mencionados asociados con la disminución del gasto urinario. Varios estudios en humanos y animales recomiendan el uso de la terapia de reemplazo renal, especialmente en aquellos que ya tienen ERC y que padecen síndrome urémico. Este informe tiene como objetivo demostrar que el uso de hemodiálisis intermitente (HI) en ovejas es una alternativa terapéutica en aquellos con insuficiencia renal. Una oveja Bergamasca de seis años fue apresentadaen decúbito lateral, disnea, diarrea líquida oscura, apatía, anorexia y crepitación pulmonar en la auscultación. Las pruebas de laboratorio mostraron alteraciones de azotemia y electrolitos como hipocalcemia, hipocalemia, hipocloremia, isostenuria (1,012), piuria, glucosuria y una relación proteína: creatinina de 0.88. En la ecografía, el índice de resistividad real fue superior a 0,54, lo que sugiere una pérdida de perfusión renal. El tratamiento elegido consistió en reemplazo electrolítico y de fluidos, asociado con HI. Después de HI,la oveja mostró una gran mejoría clínica y de laboratorio como se esperaba, sin embargo, a pesar de no haber sobrevivido, los hallazgos histopatológicos de la necropsia señalaron segmentos de glomeruloesclerosis, lo que refuerza la hipótesis de que el animal ya tenía una ERC que se agravó debido a la asociación de hipovolemia, hipotensión por fármacos anestésicos y el uso indiscriminado de AINE. A pesar de la muerte de la oveja, HI nuevamente demostró ser una terapia de rápida mejora clínica y de laboratorio del paciente en comparación con el tratamiento clínico convencional...(AU)

Assessment of neuropharmacological potential of low molecular weight components extracted from Rhinella schneideri toad poison

Background:Studies on toad poison are relevant since they are considered a good source of toxins that act on different biological systems. Among the molecules found in the toad poison, it can be highlighted the cardiotonic heterosides, which have a known mechanism that inhibit Na+/K+-ATPase enzyme. However, these poisons have many other molecules that may have important biological actions. Therefore, this work evaluated the action of the low molecular weight components from Rhinella schneideri toad poison on Na+/K+-ATPase and their anticonvulsive and / or neurotoxic effects, in order to detect molecules with actions of biotechnological interest.Methods:Rhinella schneideri toad (male and female) poison was collected by pressuring their parotoid glands and immediately dried and stored at -20 °C. The poison was dialysed and the water containing the low molecular mass molecules (< 8 kDa) that permeate the dialysis membrane was collected, frozen and lyophilized, resulting in the sample used in the assays, named low molecular weight fraction (LMWF). Na+/K+ ATPase was isolated from rabbit kidneys and enzyme activity assays performed by the quantification of phosphate released due to enzyme activity in the presence of LMWF (1.0; 10; 50 and 100 µg/mL) from Rhinella schneideri poison. Evaluation of the L-Glutamate (L-Glu) excitatory amino acid uptake in brain-cortical synaptosomes of Wistar rats was performed using [3H]L-glutamate and different concentration of LMWF (10-5 to 10 µg/µL). Anticonvulsant assays were performed using pentylenetetrazole (PTZ) and N-methyl-D-aspartate (NMDA) to induce seizures in Wistar rats (n= 6), which were cannulated in the lateral ventricle and treated with different concentration of LMWF (0.25; 0.5; 1.0; 2.0; 3.0 and 4.0 µg/µL) 15 min prior to the injection of the seizure agent.Results:LMWF induced a concentration-dependent inhibition of Na+/K+-ATPase (IC50% = 107.5 μg/mL)...(AU)

Assessment of neuropharmacological potential of low molecular weight components extracted from Rhinella schneideri toad poison

Studies on toad poison are relevant since they are considered a good source of toxins that act on different biological systems. Among the molecules found in the toad poison, it can be highlighted the cardiotonic heterosides, which have a known mechanism that inhibit Na+/K+-ATPase enzyme. However, these poisons have many other molecules that may have important biological actions. Therefore, this work evaluated the action of the low molecular weight components from Rhinella schneideri toad poison on Na+/K+-ATPase and their anticonvulsive and / or neurotoxic effects, in order to detect molecules with actions of biotechnological interest. Methods: Rhinella schneideri toad (male and female) poison was collected by pressuring their parotoid glands and immediately dried and stored at -20 °C. The poison was dialysed and the water containing the low molecular mass molecules (< 8 kDa) that permeate the dialysis membrane was collected, frozen and lyophilized, resulting in the sample used in the assays, named low molecular weight fraction (LMWF). Na+/K+ ATPase was isolated from rabbit kidneys and enzyme activity assays performed by the quantification of phosphate released due to enzyme activity in the presence of LMWF (1.0; 10; 50 and 100 µg/mL) from Rhinella schneideri poison. Evaluation of the L-Glutamate (L-Glu) excitatory amino acid uptake in brain-cortical synaptosomes of Wistar rats was performed using [3H]L-glutamate and different concentration of LMWF (10-5 to 10 µg/µL). Anticonvulsant assays were performed using pentylenetetrazole (PTZ) and N-methyl-D-aspartate (NMDA) to induce seizures in Wistar rats (n= 6), which were cannulated in the lateral ventricle and treated with different concentration of LMWF (0.25; 0.5; 1.0; 2.0; 3.0 and 4.0 µg/µL) 15 min prior to the injection of the seizure agent. Results: LMWF induced a concentration-dependent inhibition of Na+/K+-ATPase (IC50% = 107.5 μg/mL). The poison induces an increased uptake of the amino acid L-glutamate in brain-cortical synaptosomes of Wistar rats. This increase in the L-glutamate uptake was observed mainly at the lowest concentrations tested (10-5 to 10-2 µg/µL). In addition, this fraction showed a very relevant central neuroprotection on seizures induced by PTZ and NMDA. Conclusions: LMWF from Rhinella schneideri poison has low molecular weight compounds, which were able to inhibit Na+/K+-ATPase activity, increase the L-glutamate uptake and reduced seizures induced by PTZ and NMDA. These results showed that LMWF is a rich source of components with biological functions of high medical and scientific interest.(AU)

Identification of a pore-forming protein from sea anemone Anthopleura dowii Verrill (1869) venom by mass spectrometry

Background:Pore-forming proteins (PFP) are a class of toxins abundant in the venom of sea anemones. Owing to their ability to recognize and permeabilize cell membranes, pore-forming proteins have medical potential in cancer therapy or as biosensors. In the present study, we showed the partial purification and sequencing of a pore-forming protein from Anthopleura dowii Verrill (1869). 17.Methods:Cytolytic activity of A. dowii Verrill (1869) venom was determined via hemolysis assay in the erythrocytes of four mammals (sheep, goat, human and rabbit). The cytotoxic activity was analyzed in the human adherent lung carcinoma epithelial cells (A549) by the cytosolic lactate dehydrogenase (LDH) assay, and trypan blue staining. The venom was fractionated via ammonium sulfate precipitation gradient, dialysis, and ion exchange chromatography. The presence of a pore-forming protein in purified fractions was evaluated through hemolytic and cytotoxic assays, and the activity fraction was analyzed using the percent of osmotic protections after polyethylene glycol (PEG) treatment and mass spectrometry. 18.Results:The amount of protein at which the venom produced 50% hemolysis (HU50) was determined in hemolysis assays using erythrocytes from sheep (HU50 = 10.7 ± 0.2 μg), goat (HU50 = 13.2 ± 0.3 μg), rabbit (HU50 = 34.7 ± 0.5 μg), and human (HU50 = 25.6 ± 0.6 μg). The venom presented a cytotoxic effect in A549 cells and the protein amount present in the venom responsible for producing 50% death (IC50) was determined using a trypan blue cytotoxicity assay (1.84 ± 0.40 μg/mL). The loss of membrane integrity in the A549 cells caused by the venom was detected by the release of LDH in proportion to the amount of protein. The venom was fractionated; and the fraction with hemolytic and cytotoxic activities was analyzed by mass spectrometry. A pore-forming protein was identified. The cytotoxicity in the A549 cells produced by the fraction...(AU)

Validação de metodologia analítica para determinação de endotoxina bacteriana em água para diálise por meio de método cromogênico cinético / Analytical methodology validation for determining the bacterial endotoxin in the dialysis water by means of kinetic chromogenic technique

O nível de endotoxina presente na água tratada para hemodiálise é um importante indicador de qualidade, uma vez que altas concentrações de endotoxina atuam como a principal fonte de inflamação crônica em pacientes submetidos à diálise. Este estudo visa validar o método analítico para determinar quantitativamente a endotoxina bacteriana em amostras de água de hemodiálise pelo método cromogênico cinético e de comparar com o método de coagulação em gel. Os ensaios pelo método de coagulação em gel foram realizados de acordo com a Farmacopeia Brasileira em três amostras de água de hemodiálise. A validação do método cromogênico cinético foi realizada utilizando-se as mesmas amostras por meio de sistema de teste portátil. As médias geométricas das concentrações dos pontos finais obtidos nos testes de confirmação de sensibilidade LAL e de interferência por método de coagulação em gel apresentaram resultado de 0,125 UE/mL. Os resultados obtidos pelo método cromogênico para a recuperação do controle positivo do produto variaram de 89 a 186% e o coeficiente de variação de 2,5 a 18,2%, demonstrando que as amostras não apresentaram interferência. Foram obtidos resultados equivalentes em ambos os métodos, o que permite a implementação do método em laboratórios de saúde pública.

The occurrence of endotoxin in the treated water for hemodialysis is an important indicator of quality, since high concentrations of endotoxin constitute the main source for causing chronic inflammation in patients undergoing dialysis. This study aims at validating the analytical method for determining quantitatively the bacterial endotoxin in hemodialysis water samples. The data from the kinetic chromogenic method were compared with the results obtained from the gel coagulation technique. The gel coagulation assays were performed in three samples of hemodialysis water, according to the Brazilian Pharmacopoeia. The validation of the kinetic chromogenic method was performed using the same samples through the portable test system. The geometric means of the concentrations of the endpoints obtained from the tests for confirming the LAL sensitivity and the interference by gel coagulation method showed a result of 0.125 EU/mL. The results obtained by the chromogenic method for recovering the product positive control varied from 89 to 186% and the coefficient of variation from 2.5 to 18.2%, demonstrating that the samples did not show interference. Equivalent results were obtained in both methods, therefore being viable the implementation of this methodology in the public health laboratories.

Validação de metodologia analítica para determinação de endotoxina bacteriana em água para diálise por meio de método cromogênico cinético / Analytical methodology validation for determining the bacterial endotoxin in the dialysis water by means of kinetic chromogenic technique

O nível de endotoxina presente na água tratada para hemodiálise é um importante indicador de qualidade, uma vez que altas concentrações de endotoxina atuam como a principal fonte de inflamação crônica em pacientes submetidos à diálise. Este estudo visa validar o método analítico para determinar quantitativamente a endotoxina bacteriana em amostras de água de hemodiálise pelo método cromogênico cinético e de comparar com o método de coagulação em gel. Os ensaios pelo método de coagulação em gel foram realizados de acordo com a Farmacopeia Brasileira em três amostras de água de hemodiálise. A validação do método cromogênico cinético foi realizada utilizando-se as mesmas amostras por meio de sistema de teste portátil. As médias geométricas das concentrações dos pontos finais obtidos nos testes de confirmação de sensibilidade LAL e de interferência por método de coagulação em gel apresentaram resultado de 0,125 UE/mL. Os resultados obtidos pelo método cromogênico para a recuperação do controle positivo do produto variaram de 89 a 186% e o coeficiente de variação de 2,5 a 18,2%, demonstrando que as amostras não apresentaram interferência. Foram obtidos resultados equivalentes em ambos os métodos, o que permite a implementação do método em laboratórios de saúde pública.(AU)

The occurrence of endotoxin in the treated water for hemodialysis is an important indicator of quality, since high concentrations of endotoxin constitute the main source for causing chronic inflammation in patients undergoing dialysis. This study aims at validating the analytical method for determining quantitatively the bacterial endotoxin in hemodialysis water samples. The data from the kinetic chromogenic method were compared with the results obtained from the gel coagulation technique. The gel coagulation assays were performed in three samples of hemodialysis water, according to the Brazilian Pharmacopoeia. The validation of the kinetic chromogenic method was performed using the same samples through the portable test system. The geometric means of the concentrations of the endpoints obtained from the tests for confirming the LAL sensitivity and the interference by gel coagulation method showed a result of 0.125 EU/mL. The results obtained by the chromogenic method for recovering the product positive control varied from 89 to 186% and the coefficient of variation from 2.5 to 18.2%, demonstrating that the samples did not show interference. Equivalent results were obtained in both methods, therefore being viable the implementation of this methodology in the public health laboratories.(AU)

Qualificação do transporte de amostras para o monitoramento da qualidade de água tratada para uso em diálise / Qualification of the sample transport for monitoring the quality of water for dialysis purpose

As condições de coleta e o transporte das amostras de água tratada para uso em diálise são etapas críticas para garantir a qualidade dos resultados analíticos. A temperatura no transporte de amostras deve assegurar a manutenção das características físicas e biológicas desde a coleta até o recebimento pelo laboratório analítico. O objetivo deste estudo foi de qualificar o transporte de amostras de água tratada para diálise desde os Serviços de Diálise do estado de São Paulo até o Instituto Adolfo Lutz, realizado durante a execução do Programa Estadual de Monitoramento. Os resultados obtidos indicaram que o sistema adotado, utilizando-se caixas isotérmicas e gelo reutilizável, é capaz de garantir as condições adequadas de temperatura durante o transporte do ponto de coleta das amostras até o laboratório analítico.

The conditions for collecting and transporting the dialysis water samples are the critical steps to assure the quality of the analytical results. The temperature during the water samples transport should ensure the maintenance of the physical and biological characteristics of samples from their collections to the analytical laboratory. The objective of this study was to qualify the transport of the dialysis water samples from the Dialysis Services of the state of São Paulo to the Adolfo Lutz Institute, during the performance of the State Monitoring Program. By using the isothermal boxes and the reusable ices, the obtained results indicated that the adopted system were able to guarantee the adequate temperature conditions during the transport from the sample collection point to the analytical laboratory.

Qualificação do transporte de amostras para o monitoramento da qualidade de água tratada para uso em diálise / Qualification of the sample transport for monitoring the quality of water for dialysis purpose

As condições de coleta e o transporte das amostras de água tratada para uso em diálise são etapas críticas para garantir a qualidade dos resultados analíticos. A temperatura no transporte de amostras deve assegurar a manutenção das características físicas e biológicas desde a coleta até o recebimento pelo laboratório analítico. O objetivo deste estudo foi de qualificar o transporte de amostras de água tratada para diálise desde os Serviços de Diálise do estado de São Paulo até o Instituto Adolfo Lutz, realizado durante a execução do Programa Estadual de Monitoramento. Os resultados obtidos indicaram que o sistema adotado, utilizando-se caixas isotérmicas e gelo reutilizável, é capaz de garantir as condições adequadas de temperatura durante o transporte do ponto de coleta das amostras até o laboratório analítico.(AU)

The conditions for collecting and transporting the dialysis water samples are the critical steps to assure the quality of the analytical results. The temperature during the water samples transport should ensure the maintenance of the physical and biological characteristics of samples from their collections to the analytical laboratory. The objective of this study was to qualify the transport of the dialysis water samples from the Dialysis Services of the state of São Paulo to the Adolfo Lutz Institute, during the performance of the State Monitoring Program. By using the isothermal boxes and the reusable ices, the obtained results indicated that the adopted system were able to guarantee the adequate temperature conditions during the transport from the sample collection point to the analytical laboratory.(AU)

Identification, expression and characterization of the recombinant Sol g 4. 1 protein from the venom of the tropical fire ant Solenopsis geminata

Fire ant venom is a complex mixture consisting of basic piperidine alkaloids, various biologically active peptides and protein components, including a variety of major allergenic proteins. Tropical fire ant Solenopsis geminata is an important stinging ant species that causes anaphylaxis and serious medical problems. Although the biological activities of allergenic venom proteins that are unique to ant venom, particularly Solenopsis 2 and 4, are still unknown, these proteins are believed to play important roles in mediating the effects of the piperidine derivatives in the venom. Methods: In the present study, the cDNA cloning, sequencing and three-dimensional structure of Sol g 4.1 venom protein are described. The recombinant Sol g 4.1 protein (rSol g 4.1) was produced in E. coli , and its possible function as a hydrophobic binding protein was characterized by paralyzing crickets using the 50% piperidine dose (PD50). Moreover, an antiserum was produced in mice to determine the allergenic properties of Sol g 4.1, and the antiserum was capable of binding to Sol g 4.1, as determined by Western blotting. Results: The molecular weight of Sol g 4.1 protein is 16 kDa, as determined by SDS-PAGE. The complete cDNA is 414 bp in length and contains a leader sequence of 19 amino acids. The protein consists of six cysteines that presumably form three disulfide bonds, based on a predicted three-dimensional model, creating the interior hydrophobic pocket and stabilizing the structure. The rSol g 4.1 protein was expressed in inclusion bodies, as determined by SDS-PAGE. Dialysis techniques were used to refold the recombinant protein into the native form. Its secondary structure, which primarily consists of α-helices, was confirmed by circular dichroism analysis, and the three-dimensional model was also verified. The results of allergenic analysis performed on mice showed that the obtained protein was predicted to be allergenically active. Moreover, we report on the possible role of the Sol g 4.1 venom protein, which significantly reduced the PD50 from 0.027 to 0.013% in paralyzed crickets via synergistic effects after interactions with piperidine alkaloids. Conclusions: The primary structure of Sol g 4.1 showed high similarity to that of venom proteins in the Solenopsis 2 and 4 family. Those proteins are life-threatening and produce IgE-mediated anaphylactic reactions in allergic individuals. The possible function of this protein is the binding of the interior hydrophobic pockets with piperidine alkaloids, as determined by the analysis of the structural model and PD50 test.(AU)

Identification, expression and characterization of the recombinant Sol g 4.1 protein from the venom of the tropical fire ant Solenopsis geminata

Background: Fire ant venom is a complex mixture consisting of basic piperidine alkaloids, various biologically active peptides and protein components, including a variety of major allergenic proteins. Tropical fire ant Solenopsis geminata is an important stinging ant species that causes anaphylaxis and serious medical problems. Although the biological activities of allergenic venom proteins that are unique to ant venom, particularly Solenopsis 2 and 4, are still unknown, these proteins are believed to play important roles in mediating the effects of the piperidine derivatives in the venom. Methods: In the present study, the cDNA cloning, sequencing and three-dimensional structure of Sol g 4.1 venom protein are described. The recombinant Sol g 4.1 protein (rSol g 4.1) was produced in E. coli , and its possible function as a hydrophobic binding protein was characterized by paralyzing crickets using the 50% piperidine dose (PD50). Moreover, an antiserum was produced in mice to determine the allergenic properties of Sol g 4.1, and the antiserum was capable of binding to Sol g 4.1, as determined by Western blotting. Results: The molecular weight of Sol g 4.1 protein is 16 kDa, as determined by SDS-PAGE. The complete cDNA is 414 bp in length and contains a leader sequence of 19 amino acids. The protein consists of six cysteines that presumably form three disulfide bonds, based on a predicted three-dimensional model, creating the interior hydrophobic pocket and stabilizing the structure. The rSol g 4.1 protein was expressed in inclusion bodies, as determined by SDS-PAGE. Dialysis techniques were used to refold the recombinant protein into the native form. Its secondary structure, which primarily consists of -helices, was confirmed by circular dichroism analysis, and the three-dimensional model was also verified. The results of allergenic analysis performed on mice showed that the...(AU)

Identification, expression and characterization of the recombinant Sol g 4.1 protein from the venom of the tropical fire ant Solenopsis geminata

Background: Fire ant venom is a complex mixture consisting of basic piperidine alkaloids, various biologically active peptides and protein components, including a variety of major allergenic proteins. Tropical fire ant Solenopsis geminata is an important stinging ant species that causes anaphylaxis and serious medical problems. Although the biological activities of allergenic venom proteins that are unique to ant venom, particularly Solenopsis 2 and 4, are still unknown, these proteins are believed to play important roles in mediating the effects of the piperidine derivatives in the venom. Methods: In the present study, the cDNA cloning, sequencing and three-dimensional structure of Sol g 4.1 venom protein are described. The recombinant Sol g 4.1 protein (rSol g 4.1) was produced in E. coli , and its possible function as a hydrophobic binding protein was characterized by paralyzing crickets using the 50% piperidine dose (PD50). Moreover, an antiserum was produced in mice to determine the allergenic properties of Sol g 4.1, and the antiserum was capable of binding to Sol g 4.1, as determined by Western blotting. Results: The molecular weight of Sol g 4.1 protein is 16 kDa, as determined by SDS-PAGE. The complete cDNA is 414 bp in length and contains a leader sequence of 19 amino acids. The protein consists of six cysteines that presumably form three disulfide bonds, based on a predicted three-dimensional model, creating the interior hydrophobic pocket and stabilizing the structure. The rSol g 4.1 protein was expressed in inclusion bodies, as determined by SDS-PAGE. Dialysis techniques were used to refold the recombinant protein into the native form. Its secondary structure, which primarily consists of -helices, was confirmed by circular dichroism analysis, and the three-dimensional model was also verified. The results of allergenic analysis performed on mice showed that the...

Anthelmintic effect of Cassia fistula and Combretum leprosum protein fractions against goat gastrointestinal nematodes / Efeito antihelmíntico das frações proteicas de Cassia fistula e Combretum leprosum contra nematodeos gastrintestinais de caprinos

In this study, we evaluated the ovicidal and larvicidal activity of protein preparations obtained from Cassia fistula L. and Combretum leprosum Mart. leaves on the gastrointestinal parasites of goats. Protein preparations were obtained after the extraction of C. fistula L. and C. leprosum Mart. leaves, followed by protein fractionation (with ammonium sulfate saturation percentages of 30%, 30%-60%, and 60%-90%) and dialysis, which resulted in protein fractions (called F1, F2, and F3, respectively). The fractions were evaluated by egg hatching (the eggs were recovered in stool samples from naturally infected goats) and larval development tests. The results reveled that the inhibition of hatching of eggs caused by the protein fractions of C. fistula (38%) were similar to that of the control drug, thiabendazole. In addition, the fractions of C. fistula caused significant inhibition (61-69%) of larval development also. However, C. leprosum did not reveal significant inhibition of egg hatching and larval development. We conclude that C. fistula L. showed better ovicidal and larvicidal activity against endoparasites.(AU)

Neste estudo, foram avaliadas as atividades ovicida e larvicida de preparações proteicas de Cassia fistula L. e Combretum leprosum Mart. em parasitas gastrointestinais de caprinos. As preparações proteicas foram obtidas por extração das folhas de C. fistula L. e C. leprosum Mart. seguido pelo fracionamento proteico (com porcentagens de saturação de sulfato de amônio de 30%, 30-60%, 60-90%) e diálise, resultando nas frações proteicas (intituladas F1, F2 e F3, respectivamente). As frações foram avaliadas nos testes de eclosão de ovos (os ovos foram recuperados em amostras de fezes de cabras naturalmente infectadas) e de desenvolvimento larvar. Os resultados revelaram que a inibição da eclosão de ovos causada pelas frações proteicas de C. fistula (38%) foi semelhante à do fármaco controle, o tiabendazol. Além disso, as frações de C. fistula também causaram inibição significativa (61-69%) do desenvolvimento larvar. No entanto, C. leprosum não revelou inibição significativa na eclosão dos ovos e no desenvolvimento larvar. Concluiu-se que C. fistula L. mostrou uma melhor atividade ovicida e larvicida contra endoparasitas.(AU)

Detection of microorganisms, endotoxins and aluminum in mobile dialysis services / Detecção de microrganismos, endotoxinas e alumínio em serviços de diálise móvel

Hemodialysis water and dialysates are fundamental in the treatment of kidney disease.During just one hemodialysis session, 120 liters of dialysate are consumed. Thus, it is essential that theparameters of chemical and microbiological quality of the fluids used in dialysis systems are carefullyobserved. In this study, water samples were collected at 12 hospitals in the state of Rio de Janeiro. Thesamples were obtained at three points of fluid reservoirs: pre-, post-osmosis and dialysis solution. Aftercollection, colony forming units (CFU), total coliforms and Escherichia coli 100 mL-1 were quantified. Later,isolated colonies and endotoxin content were identified by biochemical assays. Data about total aluminumlevels per sample (mg L-1) were also obtained. Samples of all mobile dialysis services and points ofcollection were contaminated above the levels set out by national laws, in particular by Pseudomonasaeruginosa. Endotoxin levels were also above the recommended by current legislation (> 0.25 EU mL-1).Only three samples contained detectable levels of aluminum, which were found to be above therecommended values for the corresponding resolution (0.01 mg L-1). Finally, there were no observableamounts of total coliforms and E. coli 100 mL-1 sample. The data from this study are an important step forward in the standardization and control of chemical/microbiological quality of mobile dialysis services.(AU)

Águas de hemodiálise e dialisatos são peças fundamentais na terapêutica da doença renal.Durante apenas uma sessão de hemodiálise, são consumidos aproximadamente 120 litros de dialisato. Destaforma, é essencial que os parâmetros de qualidade microbiológica e química dos fluidos utilizados emsistemas de diálise sejam cuidadosamente observados. Neste trabalho, foram coletadas amostras de água em12 hospitais do Estado do Rio de Janeiro. Amostras foram obtidas em pontos pré-osmose, pós-osmose esolução de diálise. Após coleção, quantificaram-se unidades formadoras de colônias (UFC), coliformestotais e Escherichia coli 100 mL-1. Posteriormente, colônias isoladas e teor de endotoxinas foram identificadospor ensaios bioquímicos. Dados acerca dos níveis totais de alumínio por amostra (mg L-1) também foramobtidos. Amostras de todos os serviços de diálise móvel e pontos de coleção apresentaram contaminaçãoacima dos níveis previstos em legislação nacional, em especial por Pseudomonas aeruginosa. Teores deendotoxinas também se mostraram acima da legislação vigente (> 0,25 EU mL-1). Apenas três amostrascontinham níveis detectáveis e acima dos valores preconizados por resolução correspondente (0,01 mg L-1).Por fim, não foram encontradas quantidades observáveis de coliformes totais e E. coli 100 mL-1 de amostra.Dados de nosso estudo são importante avanço na padronização e controle de qualidadequímica/microbiológica em serviços de diálise móvel.(AU)