Action of bromelain and ficin on horse anti Bothrops sp venom antibodies

The treatment with hyperimmune sera constitute the only specific and effective therapy available against snakebite envenomation, most common in developing countries. Serum quality is an important factor on patient recovery time and in the incidence of death and permanent disability. To date, most sera consist of pepsin digested IgG antibodies harvested from hyperimmune animals. The use of animal derived enzymes, such as pepsin, to digest IgG, constitute a source of adventitious agents and contaminants, such as porcine circovirus. The present study aims to evaluate the use of the plant derived enzymes bromelain and ficin, as an alternative to pepsin. To this purpose, horse serum immunized against Bothrops venoms was purified with caprylic acid and digested with bromelain or ficin. SDS-PAGE results evidence the formation of F(ab)’2 fragments and suggest that a digestion time superior to 8 hours may be required to completely digest the antibodies with bromelain or ficin. F(ab)’2 fragments obtained by digestion with either bromelain or ficin digestion preserved the ability to recognize Bothrops sp. venom in western blotting assays. Therefore, both enzymes are suitable for use in large-scale production, minimizing contamination risks and increasing safety and efficiency of serotherapy treatments.

Action of Bromelain and Ficin on horse anti Bothrops sp venom Antibodies

Abstract The treatment with hyperimmune sera constitute the only specific and effective therapy available against snakebite envenomation, most common in developing countries. Serum quality is an important factor on patient recovery time and in the incidence of death and permanent disability. To date, most sera consist of pepsin digested IgG antibodies harvested from hyperimmune animals. The use of animal derived enzymes, such as pepsin, to digest IgG, constitute a source of adventitious agents and contaminants, such as porcine circovirus. The present study aims to evaluate the use of the plant derived enzymes bromelain and ficin, as an alternative to pepsin. To this purpose, horse serum immunized against Bothrops venoms was purified with caprylic acid and digested with bromelain or ficin. SDS-PAGE results evidence the formation of F(ab)'2 fragments and suggest that a digestion time superior to 8 hours may be required to completely digest the antibodies with bromelain or ficin. F(ab)'2 fragments obtained by digestion with either bromelain or ficin digestion preserved the ability to recognize Bothrops sp. venom in western blotting assays. Therefore, both enzymes are suitable for use in large-scale production, minimizing contamination risks and increasing safety and efficiency of serotherapy treatments.

Small cell lung cancer: an overview of the targets

Abstract Lung cancer is the leading cause of cancer deaths worldwide. Small cell lung cancer (SCLC) accounts for approximately 15% of all lung cancer cases. Despite a frequently good response to first-line treatment with chemotherapy and/or radiotherapy, early relapse occurs in the majority of patients and 5-year survival is only about 5%. This histological subtype of lung cancer is strongly associated with tobacco smoking. The behavior of SCLC is unique within solid tumors. Initially, it positively responds to chemotherapy or radiotherapy. However, at relapse, which occurs early in the majority of cases, the tumor is resistant to available therapy and eventually will cause the death of the patient. These results in an overall 5-year survival of approximately 5% for the entire population of patients diagnosed with SCLC. This dismal prognosis has not significantly changed in past years. There is an urgent need for discovery targets to select patients more prone to having a proper response to the treatment, avoiding to reduce their resistance and resulting the increase of overall and progression-free survivals.

CD14 and IL6 polymorphisms are associated with a pro-atherogenic profile in young adults with acute myocardial infarction

Abstract This study investigated the relationship of polymorphismsin genes encoding CD14, IL-6 and TLR4 withmetabolic, inflammatory and endothelial markers in youngadults with acute myocardial infarction (AMI). Glucose,lipids, nitrate and inflammatory markers, flow mediatedvasodilatation (FMV) and flow mediated by nitrate (FMN)were evaluated in 102 AMI and 108 non-AMI (controlgroup) young individuals (% years). CD14 -260C[T(rs2569190), IL6 -174G[C (rs1800795) and TLR4c.896A[G (rs4986790) and TLR4 c.1196C[T (rs4986791)polymorphisms were analyzed by PCR–RFLP. Minor allelefrequencies of CD14, IL6 and TLR4 polymorphisms weresimilar between AMI and control groups (p[0.05). In AMIgroup, individuals carrying IL6 -174CC genotype hadhigher serum triglycerides, VLDL cholesterol and glucosecompared to the IL6 -174GG/GC genotype carriers(p84 2013-11-12

Biological and physicochemical stability of ceftazidime and aminophylline on glucose parenteral solution

Ceftazidime is a broad spectrum antibiotic administered mainly by the parenteral route, and it is especially effective against Pseudomonas aeruginosa. The period of time in which serum levels exceed the Minimum Inhibitory Concentration (MIC) is an important pharmacodynamic parameter for its efficacy. One of the forms to extend this period is to administer the antibiotic by continuous infusion, after prior dilution in a Parenteral Solution (PS). The present work assessed the stability of ceftazidime in 5% glucose PS for 24 hours, combined or not with aminophylline, through High Performance Liquid Chromatography (HPLC). The physicochemical evaluation was accompanied by in vitro antimicrobial activity compared MIC test in the 24-hour period. Escherichia coli and Pseudomonas aeruginosa were the microorganisms chosen for the MIC comparison. The HPLC analysis confirmed ceftazidime and aminophylline individual stability on PS, while the MIC values were slightly higher than the mean described in the literature. When both drugs were associated in the same PS, the ceftazidime concentration by HPLC decreased 25% after 24 hours. Not only did the MIC values show high loss of antibiotic activity within the same period, but also altered MIC values immediately after the preparation, which was not detected by HPLC. Our results indicate that this drug combination is not compatible, even if used right away, and that PS might not be the best vehicle for ceftazidime, emphasizing the importance of the MIC evaluation for drug interactions.

Ceftazidima é um antimicrobiano administrado por via parenteral, que apresenta amplo espectro de ação, principalmente contra Pseudomonas aeruginosa. O tempo em que a concentração sérica de ceftazidima permanece acima da concentração mínima inibitória (MIC) é um importante parâmetro farmacodinâmico para a determinação da eficácia antimicrobiana e pode ser potencializado através da utilização de infusão contínua em soluções parenterais (PS). Este artigo visa a avaliar a estabilidade da ceftazidima em solução de glicose 5%, na presença e na ausência do fármaco aminofilina, através de cromatografia líquida de alta eficiência HPLC e MIC durante o período de 24 horas. Os microorganismos selecionados para a determinação do MIC foram Escherichia coli e Pseudomonas aeruginosa. Os ensaios em cromatógrafo líquido confirmaram a estabilidade dos fármacos ceftazidima e aminofilina quando são individualmente associados em PS, enquanto os valores de MIC ficaram maiores que os valores encontrados na literatura. Quando ambos os fármacos foram associados na mesma solução parenteral a concentração de ceftazidima obtida por HPLC diminuiu 25% depois de 24 horas. Os valores de MIC mostraram maior decaimento da atividade antimicrobiana neste mesmo período e também valores de MIC alterados nas soluções preparadas no tempo zero, decaimento este que não foi detectado em HPLC. Os resultados indicaram incompatibilidade na associação dos fármacos em PS, enfatizando a importância dos resultados de MIC para interações de fármacos.

Immunogenicity of a Whole-Cell Pertussis Vaccine with Low Lipopolysaccharide Content in Infants

The lack of a clear correlation between the levels of antibody to pertussis antigens and protection against disease lends credence to the possibility that cell-mediated immunity provides primary protection against disease...

Veneno de Bothrops jararaca modificado com PEG (monometoxipolietileno glicol) para elaboração de um adjuvante complexado à partícula antigênica / Bothrops jararaca venon modified for elaboration of complexed adjuvant to the antigenic particule with (monomethoxypolyethylene glycol (mPEG)

Enzimas, biofármacos e produtos de origem biológica de modo geral sempre foram obstáculos para o uso terapêutico. Isto em função dos riscos de reação adversa que poderiam ocasionar bem como a indução de uma resposta imunológica desejável ou não. A utilização do monometoxipolietileno glicol (mPEG) conjugado a proteínas e enzimas trouxe novas perspectivas de utilização de substâncias naturais como substâncias farmacológicas ativas, pois o fato de estarem conjugadas a um polímero inerte atribui uma nova estrutura físico-química a essas substâncias, diminuindo consideravelmente os riscos de anafilaxia e a biodegradação por formação de complexos antígeno-anticorpo...

Effect of beta-propiolactone treatment on the complement activation mediated by equine antisera

A reducao da ativacao do complemento atraves de uma alteracao do fragmento Fc das imunoglobulinas pela beta-propiolactona foi obtida em soros hiperimunes equinos antivirus rabico, venenos Bothrops e toxina difterica. Os resultados foram avaliados por teste de anafilaxia em cobaias, e comparados com aqueles obtidos com os mesmos soros purificados por precipitacao salina (sulfato de amonio), seguidos ou nao por digestao enzimatica com pepsina. Os niveis de pureza proteica foram para o soro antibotropico de 184,5 mg/g e 488,5 mg/g tratado pela beta-propiolactona e digeridos pela pepsina, respectivamente...(au)

Produçäo de anticorpos antiveneno total de Crotalus durissus terrificus em cavalos por fosfolipase A2 / Antigens antivenom's total production of Crotalus durissus terrificus in horses by phospholipase A2

Fosfolipases A2 purificada de veneno de Crotalus durissus terrificus ou veneno integral foram usados, como antígenos, para imunizar cavalos e burros. A imunizaçäo de base foi feita injetando os animais, pela via subcutânea, com 5mg do antígeno emulsionado em adjuvante de Freund completo. Quatro meses depois, os animais foram reimunizados injetando-se os antígenos correspondentes, dissolvidos em salina, ao redor dos granulomas resultantes da imunizaçäo primária. Esta injeçäo foi repetida por mais cinco vezes a intervalos de uma semana..

Cross - reactivity of horse monovalent antivenoms to venoms of ten bothrops species

Horses were immunized with B. alternatus, B. atrox, B. cotiara, B. erythromelas, B. insularis, B. jararaca, B. jararacussu, B, moojeni, B. neuwiedi and B. pradoi venoms. Antibodies recognizing the venom antigenic components were either immunochemically detected by the precipattion methods or biologically by the assays measuring the venomns indirect hemolytic and lethal toxic activities . Specific and cross - reacting antivenoms.Modifications in the serum electrophoretic patterns characterized by a reduction of the albumin peak and by a correspondent increase of the y-globulins with a pattent or no modification of the α or β globulins were found in these sera.

Cavalos foram imunizados com veneno de B. alternatus, B. atrox, B. cotiara, B. erythromelas, B. insularis, B. jararaca, B. jararacussu, B, moojeni, B. neuwiedi and B. pradoi. Anticorpos específicos para componentes antigênicos dos venenos foram detectados pelos métodos imuno- enzimático, dupla-difusão e precipitação quantitativa enquanto que os anticorpos neutralizantes foram analisados pelos métodos da hemólise indireta em placas e pela neutralização de seus efeitos letais. Anticorpos, tanto específicos como dando reações cruzadas com venenos botrópicos foram encontrados em todos os dez soros monovalentes. Modificações nos padrões eletroforéticos, caracterizadas por uma redução no pico da albumina e por um correspondente aumento das γ. globulinas com modificações ora acentuadas ora pouco perceptíveis nas frações das α e β globolinas, foram detectadas todos esses soros.