Vitamins as influenza vaccine adjuvant components.

A number of adjuvant formulations were assayed in mice immunized with 3.75 µg of A/California/7/2009 (H1N1) pdm09 influenza vaccine with vitamins A, D and/or E in emulsions or B2 and/or B9 combined with Bordetella pertussis MPLA and/or alum as adjuvants. Squalene was used as positive control, as well as MPLA with alum. The immune response was evaluated by a panel of tests, including a hemagglutination inhibition (HAI) test, ELISA for IgG, IgG1, and IgG2a and IFN-γ, IL-2, IL-6 and IL-10 quantification in splenocyte culture supernatant after stimulus with influenza antigen. Immunological memory was evaluated using a 1/10 dose booster 60 days after the first immunization followed by assessment of the response by HAI, IgG ELISA, and determination of the antibody affinity index. The highest increases in HAI, IgG1 and IgG2a titers were obtained with the adjuvant combinations containing vitamin E, or the hydrophilic combinations containing MPLA and alum or B2 and alum. The IgG1/IgG2a ratio indicates that the response to the combination of B2 with alum would have more Th2 character than the combination of MPLA with alum. In an assay to investigate the memory response, a significant increase in HAI titer was observed with a booster vaccine dose at 60 days after immunization with vaccines containing MPLA with alum or B2 with alum. Overall, of the 27 adjuvant combinations, MPLA with alum and B2 with alum were the most promising adjuvants to be evaluated in humans.

Surfactant protein A is decreased in the lung of rats with hepatopulmonary syndrome.

PURPOSE: To evaluate surfactant protein A levels in an hepatopulmonary syndrome rat model. To date, there have been no studies aimed at evaluating surfactant levels in the setting of cirrhosis or hepatopulmonary syndrome. METHODS: A total of 35 rats were divided into control, sham, and experimental HPS groups. We evaluated surfactant protein A levels in rats and the experimental model designed to induce hepatopulmonary syndrome was common bile duct ligation. Statistical analysis was performed using GraphPad Prism Software(r). Differences were considered statistically significant when p<0.05. RESULTS: Lung homogenate of surfactant protein A levels were lower in the experimental hepatopulmonary syndrome and sham groups in comparison to the control group (p<0.05). Serum SP-A levels were the same in experimental hepatopulmonary syndrome and control groups but decreased in the sham group compared with the experimental groups (p<0.05). Myeloperoxidase activity was higher in the experimental hepatopulmonary syndrome group than the other two groups (p<0.05). CONCLUSION: Surfactant protein A is present in experimental hepatopulmonary syndrome and leads to an imbalance between serum and pulmonary levels due to systemic inflammatory response.

Surfactant protein A is decreased in the lung of rats with hepatopulmonary syndrome

PURPOSE: To evaluate surfactant protein A levels in an hepatopulmonary syndrome rat model. To date, there have been no studies aimed at evaluating surfactant levels in the setting of cirrhosis or hepatopulmonary syndrome. METHODS: A total of 35 rats were divided into control, sham, and experimental HPS groups. We evaluated surfactant protein A levels in rats and the experimental model designed to induce hepatopulmonary syndrome was common bile duct ligation. Statistical analysis was performed using GraphPad Prism Software(r). Differences were considered statistically significant when p<0.05. RESULTS: Lung homogenate of surfactant protein A levels were lower in the experimental hepatopulmonary syndrome and sham groups in comparison to the control group (p<0.05). Serum SP-A levels were the same in experimental hepatopulmonary syndrome and control groups but decreased in the sham group compared with the experimental groups (p<0.05). Myeloperoxidase activity was higher in the experimental hepatopulmonary syndrome group than the other two groups (p<0.05). CONCLUSION: Surfactant protein A is present in experimental hepatopulmonary syndrome and leads to an imbalance between serum and pulmonary levels due to systemic inflammatory response. .

Surfactant protein A is decreased in the lung of rats with hepatopulmonary syndrome

PURPOSE:To evaluate surfactant protein A levels in an hepatopulmonary syndrome rat model. To date, there have been no studies aimed at evaluating surfactant levels in the setting of cirrhosis or hepatopulmonary syndrome.METHODS:A total of 35 rats were divided into control, sham, and experimental HPS groups. We evaluated surfactant protein A levels in rats and the experimental model designed to induce hepatopulmonary syndrome was common bile duct ligation. Statistical analysis was performed using GraphPad Prism Software(r). Differences were considered statistically significant when p<0.05.RESULTS:Lung homogenate of surfactant protein A levels were lower in the experimental hepatopulmonary syndrome and sham groups in comparison to the control group (p<0.05). Serum SP-A levels were the same in experimental hepatopulmonary syndrome and control groups but decreased in the sham group compared with the experimental groups (p<0.05). Myeloperoxidase activity was higher in the experimental hepatopulmonary syndrome group than the other two groups (p<0.05).CONCLUSION:Surfactant protein A is present in experimental hepatopulmonary syndrome and leads to an imbalance between serum and pulmonary levels due to systemic inflammatory response.(AU)

An improved whole cell pertussis vaccine with reduced content of endotoxin.

An improved whole cell pertussis vaccine, designated as Plow, which is low in endotoxicity due to a chemical extraction of lipo-oligosaccharide (LOS) from the outer membrane, was evaluated for safety, immunogenicity and potency, comparatively to a traditional whole cell pertussis vaccine. Current whole cell pertussis vaccines are effective but contain large quantities of endotoxin and consequently display local and systemic adverse reactions after administration. Endotoxin is highly inflammatory and contributes considerably to the reactogenicity as well as the potency of these vaccines. In contrast, acellular pertussis vaccines hardly contain endotoxin and are significantly less reactogenic, but their elevated costs limit their global use, especially in developing countries. In this paper, bulk products of Plow and a traditional whole cell vaccine, formulated as plain monocomponents or combined with diphtheria and tetanus toxoids (DTPlow or DTP, respectively) were compared by in vitro and in vivo assays. Chemical extraction of LOS resulted in a significant decrease in endotoxin content (20%) and a striking decline in endotoxin related toxicity (up to 97%), depending on the used in vitro or in vivo test. The LOS extraction did not affect the integrity of the product and, more importantly, did not affect the potency and/or stability of DTPlow. Moreover, hardly any differences in antibody and T-cell responses were observed. The development of Plow is a significant improvement regarding the endotoxicity of whole cell pertussis vaccines and therefore a promising and affordable alternative to currently available whole cell or acellular pertussis vaccines for developing countries.

Medicina interna de bezerros clonados: distúrbios clínicos observados nos primeiros 30 dias de vida / Internal medicine of cloned calf: clinical disorders observed during the first 30 days of life

Apesar dos esforços realizados nos últimos quatro anos, as taxas de mortalidade dos bezerros clonados da raça que chegam a termo são ainda altas, cerca de 50%. Demonstrou-se a ocorrência de graves distúrbios cardiorrespiratórios caracterizados por hiperfonese, presença de sopros cardíacos na 1ª e 2ª bulha associadosa dispneias, respiração rude e estertores. Em consequência ao nãofechamento do Forâmen de Botal e do Ducto Arterioso, há mistura de sangue arterial e venoso comprometendo a capacidade de oxigenação do sangue dos bezerros clonados. Observou-se ainda a ocorrência de macrossomia, hipoglicemia, hipotermia, anomaliasdas estruturas umbilicais, anemia e alopecia.

Despite all efforts during the last four years to improve cloned newborn care, the mortality rate of calf after term is still high, around 50%. Clinical symptoms observed in these cloned calf were related to severe cardiopulmonary disorders like hyperphonese, diastolic and systolic cardiac murmurs associated to dyspnea and crackling lung sounds. Due to the patency of the Foramen Ovale and Ductus Arteriosus, in which causes the mix of arterial and venous blood, the blood oxygenation in these cloned calf is compromised. In addition, cloned calf could also present increasedbirth weight, hypoglycemia, hypothermia, umbilical cord abnormalities, anemia, and alopecia.

Economical value of vaccines for the developing countries-the case of instituto butantan, a public institution in Brazil

A recent review has discussed the economic value of vaccine for developed countries. The situation is quite different in developing countries, and we examine the situation in Brazil. Vaccines are of fundamental importance for the control of infectious diseases, especially among the population that lives in poor sanitary conditions. Also, vaccines can generate herd effects that result in protection even among those who have not been vaccinated, which can be of particular value to poor individuals who are not reached by health services. In appreciation of this importance, various international agencies, including the Pan American Health Organization (PAHO) and UNICEF, undertake large-scale procurement of vaccines for supply to developing countries. This scale of procurement has allowed these agencies to obtain very low prices. In Brazil, the Constitution includes the right to health care, which has led the government to formulate a goal of universal vaccination free of charge, a cost-effective measure against many important infectious diseases. Universal vaccination is a fundamental role of the federal, state, and municipal governments through the current unified public health care system (Sistema Único de Sáude - SUS).

Medicina interna de bezerros clonados: distúrbios clínicos observados nos primeiros 30 dias de vida / Internal medicine of cloned calf: clinical disorders observed during the first 30 days of life

Apesar dos esforços realizados nos últimos quatro anos, as taxas de mortalidade dos bezerros clonados da raça que chegam a termo são ainda altas, cerca de 50%. Demonstrou-se a ocorrência de graves distúrbios cardiorrespiratórios caracterizados por hiperfonese, presença de sopros cardíacos na 1ª e 2ª bulha associadosa dispneias, respiração rude e estertores. Em consequência ao nãofechamento do Forâmen de Botal e do Ducto Arterioso, há mistura de sangue arterial e venoso comprometendo a capacidade de oxigenação do sangue dos bezerros clonados. Observou-se ainda a ocorrência de macrossomia, hipoglicemia, hipotermia, anomaliasdas estruturas umbilicais, anemia e alopecia.(AU)

Despite all efforts during the last four years to improve cloned newborn care, the mortality rate of calf after term is still high, around 50%. Clinical symptoms observed in these cloned calf were related to severe cardiopulmonary disorders like hyperphonese, diastolic and systolic cardiac murmurs associated to dyspnea and crackling lung sounds. Due to the patency of the Foramen Ovale and Ductus Arteriosus, in which causes the mix of arterial and venous blood, the blood oxygenation in these cloned calf is compromised. In addition, cloned calf could also present increasedbirth weight, hypoglycemia, hypothermia, umbilical cord abnormalities, anemia, and alopecia.(AU)

Clinical disorders observed during the first 30 days of life of cloned Zebu calf

Background: The advent of animal cloning from embryos reconstructed with nuclei from cells at different stages of differentiation has been responsible for the appearance of several anomalies in calves, with some alterations diagnosed during pregnancy and others after the birth of cloned animals. Despite efforts over the past five years, the rate of mortality after the birth of cloned calves is still high, at about 50%. Below is reported the research experience gained at the University of São Paulo in the period from 2005 to 2010, related to the birth of cloned calves from Nellore. Review: Research showed that cloned calves present cardiopulmonary disorders characterized by tachycardia with episodes of arrhythmia with bradycardia, increase in the 1st heart sound, presence or absence of heart murmurs in the 1st and 2nd heart sound associated with dyspnea, breathing harsh and crackling dry, moist and crackling. During the first days of life of cloned calves that died, ultrasound examination demonstrated the presence of concentric hypertrophy congenital cardiac and turbulent flow at the foramen of botal towards the right heart to left heart, indicating reversal of blood flow to the pattern observed in fetal life. As a result, mixture of arterial and venous blood occurs, compromising the ability of oxygen to the blood of cloned calves. The occurrence of macrosomia was observed in 20.9% of Nellore calves. It is believed that this syndrome is associated with disorders of carbohydrate metabolism in the placenta and/or fetus, leading to fetal gigantism, which mimicks excessive fetal growth seen in pregnant women with diabetes. We observed the occurrence of moderate to severe normocytic and normochromic anemia, with the anemia gradually appearing from 12 h of life onwards, reaching its maximum intensity at the end of the first week, to start a gradual recovery to normal values from the 15th day of life. The anemia observed in cloned calves was ferropriva, as such animals showed a significant decrease in serum iron levels associated with decreased transferrin saturation index (STI). Umbilical anomalies occurred in most clones. There was an increase in the thickness of the umbilical cord, which hindered its spontaneous rupture at birth. The umbilical arteries did not undergo retraction into the abdominal cavity, leaving them exposed in the remnant of the umbilical cord. In the first three days after birth, strong pulse of these arteries was noted in umbilical cord, making it necessary to use clamps on the arteries in order to prevent bleeding. There were also intra-abdominal hematomas involving the arteries and the urachus. Between 15 and 20 days of life, it was observed the occurrence of alopecia in about 75.0% of the calves, whose origin may be related to disturbances in the synthesis and absorption of vitamins, since the supplementation of calves with ADE vitamin complex decreased symptoms. Conclusion: The results confirm the occurrence of serious cardiopulmonary disorders characterized by tachycardia, hyperphonesis, presence of heart murmurs in the 1st and 2nd heart sounds, episodes of arrhythmia and bradycardia associated with dyspnea, rales, and coarse breath. As a result of the non-closure of the foramen of Botal and the Ductus Arteriosus, arterial and venous blood a mix, compromising the oxygen carrying capacity in the blood of cloned calves. There was also the occurrence of macrosomia, hypoglycemia, hypothermia, abnormal umbilical structures, anemia and alopecia

Purification and characterization of aprotinin from porcine lungs.

Aprotinin, the most studied serine proteinase inhibitor, was isolated from porcine lung for the first time. The purified porcine aprotinin had an Mr value of approximately 7 kDa. It cross-reacted with polyclonal serum anti-commercial aprotinin. About 1 microg porcine aprotinin inhibited 6 microg trypsin whereas 1 microg commercial soybean inhibitor inhibited only 1 microg trypsin. The aprotinin gene was also isolated from porcine lung: the deduced amino acid sequence showed 74% identity to bovine aprotinin.

Purification and characterization of aprotinin from porcine lungs

Aprotinin, the most studied serine proteinase inhibitor, was isolated from porcine lung for the first time. The purified porcine aprotinin had an Mr value of ¡­7 kDa. It cross-reacted with polyclonal serum anti-commercial aprotinin. About 1 ¥ìg porcine aprotinin inhibited 6 ¥ìg trypsin whereas 1 ¥ìg commercial soybean inhibitor inhibited only 1 ¥ìg trypsin. The aprotinin gene was also isolated from porcine lung: the deduced amino acid sequence showed 74% identity to bovine aprotinin.

Pulmonary surfactant protein A isolation as a by-product of porcine pulmonary surfactant production.

A pulmonary surfactant reduces surface tension at the air/liquid interface of the alveoli and stabilizes alveoli at low lung volumes. Surfactant deficiency and dysfunction were shown to be present in a number of pulmonary diseases, and surfactant replacement therapy is the common clinical conduct. The hydrophilic SP-A (surfactant protein A) is absent when solvent extraction was used during exogenous surfactant production. Addition of SP-A to the surfactant preparation increases the surface activity and completely counteracts inhibition by blood proteins. SP-A recognizes and binds to carbohydrate structures on the surfaces of pathogenic micro-organisms, and acts as opsonins or cross-linking molecules by binding to a variety of cells that participate in the pulmonary immune response. The purification procedure yielded 206 mg of high-purity SP-A/kg of porcine lung, as judged by gel filtration, SDS/PAGE and Western blotting. The electrophoretic profiles obtained showed that pure SP-A consists of proteins of wide molecular mass in the range 26-36 kDa and a dimer in the range 56-60 kDa. The Western-blot results displayed the same band pattern profile after incubating the membrane using a commercially available polyclonal anti-SP-A antibody produced in goat. Gel-filtration experiments confirmed the molecular mass of SP-A in 10 mM NaCl solution. The isolated SP-A showed mannose-binding ability, representative of its functionality.

Suscetibilidade da linhagem de células Vero a cepas vacinais do vírus do sarampo / Susceptibility of Vero cell line to vaccine strains of the measles vírus

A suscetibilidade da linhagem de células Vero ao vírus do sarampo é bem conhecida e sua utilizaçäo no controle da potência da vacina contra o sarampo é amplamente difundida. Com o objetivo de comparar a suscetibilidade de células Vero empregadas em titulaçöes, amostras provenientes de dois laboratórios controladores (Vero IB e Vero INCQS), foram testadas frente a três cepas vacinais: Moraten Schwarz e Biken CAM-70. Foram titulados 72 lotes de vacinas contra o sarampo, sendo 25 produzidos com a cepa Moraten, 24 com a cepa Schwarz e 23 com a cepa Biken CAM-70. A análise estatística dos resultados obtidos nas titulaçöes, feita através dos testes Limites para uma Média e "t" de Student, mostrou que para as cepas Moraten e Biken CAM-70, as diferenças de títulos näo foram estatisticamente significantes, o mesmo näo ocorrendo com a cepa Schwarz, para a qual as células Vero IB se mostraram mais sensíveis