Anti-inflammatory potential of pyocyanin in LPS-stimulated murine macrophages.

Context: Pyocyanin is a typical Pseudomonas aeruginosa virulence factor, a common Gram-negative rod responsible for a wide range of severe nosocomial infections. There is evidence indicating that pyocyanin has multiple biological activities, but little is known about anti-inflammatory properties. Objective: This study investigated pyocyanin effect on nitric oxide and cytokine production in lipopolysaccharide (LPS)-activated murine peritoneal macrophages. Materials and methods: Macrophages were incubated in the presence and absence of pyocyanin (1, 5, 10, 50, and 100 µM) with and without LPS (1 µg/mL). Nitric oxide production was determined by Griess reagent and tumor necrosis factor (TNF)-α and interleukin (IL)-1ß production was assessed by enzyme-linked immunosorbent assay. In addition, pyocyanin effects on zymosan A-induced peritonitis in mice were evaluated. Results: Pyocyanin (5 and 10 µM) decreased nitric oxide, TNF-α, and IL-1ß production independent of macrophage death. On the other hand, in vivo, pyocyanin (5 mg/kg) was not able to affect leukocyte migration into the site of inflammation. Discussion and conclusion: Thus, our findings suggest that pyocyanin exerts anti-inflammatory effects on murine peritoneal macrophages, downregulating nitric oxide, TNF-α, and IL-1ß levels, which seems to be independent of cell migration. These effects may represent a mechanism of immune evasion; nevertheless more detailed studies should be performed to confirm this hypothesis.

Effects of rumen-undegradable protein on intake, performance, and mammary gland development in prepubertal and pubertal dairy heifers.

The objective of this study was to evaluate the influence of different amounts of rumen-undegradable protein (RUP) on intake, N balance, performance, mammary gland development, carcass traits, and hormonal status of Holstein heifers at different physiological stages (PS). Sixteen prepubertal (PRE) heifers (initial BW = 106 ± 7.6 kg; age = 4.3 ± 0.46 mo) and 16 pubertal (PUB) heifers (initial BW = 224 ± 7.9 kg; age = 12.6 ± 0.45 mo) were used in an experiment over a period of 84 d. Four diets with increasing RUP contents (38, 44, 51, and 57% of dietary crude protein) and heifers at 2 PS (PRE or PUB) were used in a 4 × 2 factorial arrangement of treatments in a completely randomized design. Throughout the experiment, 2 digestibility trials were performed over 5 consecutive days (starting at d 36 and 78) involving feed and ort sampling and spot collections of feces and urine. At d 0 and 83, body ultrasound images were obtained for real-time carcass trait evaluation. The mammary gland was ultrasonically scanned at d 0 and every 3 wk during the experiment. Blood samples were taken at d 0 and 84 to determine serum concentrations of progesterone, estrogen, insulin-like growth factor I (IGF-I), and insulin. No interaction between PS and the level of RUP was found for any trait. Apparent digestibility of dry matter, organic matter, and neutral detergent fiber corrected for ash and protein was not affected by RUP level but was lower for PRE compared with PUB heifers. Sorting against neutral detergent fiber corrected for ash and protein (tendency only) and for crude protein was greater for PUB than PRE heifers. Pubertal heifers had greater average daily gain (905 vs. 505 g/d) and N retention (25.9 vs. 12.5 g/d) than PRE heifers. In addition, average daily gain and N retention were greatest at 51% RUP of dietary protein. Mammary ultrasonography indicated no effects of RUP amounts on mammary gland composition, whereas PRE heifers had greater pixel values than PUB, indicating higher contents of fat rather than protein in the mammary glands of PRE heifers. Serum progesterone and IGF-I concentration was affected only by PS, and PRE heifers had greater values of progesterone and IGF-I concentrations than PUB heifers. Serum insulin concentration was unaffected by PS but tended to be higher at 51% of RUP. In conclusion, an RUP level of 51% increases body weight, average daily gain, feed efficiency, and N retention in heifers regardless of the PS. In addition, PRE heifers have a lower sorting ability and reduced intake, total-tract digestibility, and N retention. They also have higher amounts of fat in their mammary glands, even at moderate growth rates.

Doxorubicin hinders DNA condensation promoted by the protein bovine serum albumin (BSA).

In this work, we have studied the interaction between the anticancer drug doxorubicin (doxo) and condensed DNA, using optical tweezers. To perform this task, we use the protein bovine serum albumin (BSA) in the working buffer to mimic two key conditions present in the real intracellular environment: the condensed state of the DNA and the abundant presence of charged macromolecules in the surrounding medium. In particular, we have found that, when doxo is previously intercalated in disperse DNA, the drug hinders the DNA condensation process upon the addition of BSA in the buffer. On the other hand, when bare DNA is firstly condensed by BSA, doxo is capable to intercalate and to unfold the DNA condensates at relatively high concentrations. In addition, a specific interaction between BSA and doxo was verified, which significantly changes the chemical equilibrium of the DNA-doxo interaction. Finally, the presence of BSA in the buffer stabilizes the double-helix structure of the DNA-doxo complexes, preventing partial DNA denaturation induced by the stretching forces.

Photoperiod Differences in Sand Fly (Diptera: Psychodidae) Species Richness and Abundance in Caves in Minas Gerais State, Brazil.

Caves are unique habitats that are inhabited by a diverse and singular biota. Among these inhabitants are sand flies, which are of great epidemiological interest in the Neotropical region because they are vectors of Leishmania The period of activity of these insects is usually crepuscular and nocturnal, but there are reports of diurnal activity of sand flies in caves. Thus, the aim of this study was to evaluate the periodicity of daily activity of sand flies in cave environments in the municipality of Pains, Minas Gerais. Sand flies were collected with light traps, which were operated for 5 consecutive days in the rainy season and in the dry season. Samples were collected every 12 h and separated between photophase and scotophase periods. In total, 1,777 sand flies of 23 species were collected. The most abundant species was Lutzomyia renei (Martins, Falcão, and Silva) (44%), followed by Lutzomyia longipalpis (Lutz and Neiva) (15%), Evandromyia edwardsi (Mangabeira) (11%), and Micropygomyia quinquefer (Costa Lima) (6%). The richness and abundance of total sand flies and the abundance of male and female sand flies in the aphotic zone of the caves did not differ between the photophase and scotophase, but differed between photoperiods at the entrance and at sites surrounding the caves. From our study of the daily activity of these insects in this ecotope, it will be possible to know which period of the day is of greatest risk of exposure of vertebrates who visit or live in these environments, including the human population.

[Evaluation of the pain in older people]. / Evaluación del dolor en el adulto mayor.

OBJECTIVE: To compare the information obtained of the percentage of improvement declared orally with the improvement marked in the visual analogical scale (VAS) in patients under and over 65 years of age. PATIENTS AND METHODS: Ninety-five individuals with acute shoulder pain (enthesitis) were evaluated. The subjects were requested to mark the pain intensity in the VAS before a treatment with corticosteroid injection in the shoulder and were evaluated again through the VAS one week after the procedure. They were also requested to declare orally the pain intensity. Then, the information was compared between patients aged under and over 65 years of age. RESULTS: 29.8% of those younger than 65 years, and 60.95% of those older than that age presented more than 10% difference between orally stated and calculated pain relief percentage based on the VSA. CONCLUSION: The difference between the orally stated and the calculated pain relief percentage based on the VSA was significantly higher in the group of those older than 65 years. This finding proved that the VAS is a poor method to evaluate pain relief in the senior population.

Comparar las informaciones obtenidas del porcentaje verbal de mejoría declarada verbalmente con la escala visual analógica (EVA) en personas con edad menor y mayor o igual de 65 años.

Depletion interactions and modulation of DNA-intercalators binding: Opposite behavior of the "neutral" polymer poly(ethylene-glycol).

In this work we have investigated the role of high molecular weight poly(ethylene-glycol) 8000 (PEG 8000) in modulating the interactions of the DNA molecule with two hydrophobic compounds: Ethidium Bromide (EtBr) and GelRed (GR). Both compounds are DNA intercalators and are used here to mimic the behavior of more complex DNA ligands such as chemotherapeutic drugs and proteins whose domains intercalate DNA. By means of single-molecule stretching experiments, we have been able to show that PEG 8000 strongly shifts the binding equilibrium between the intercalators and the DNA even at very low concentrations (1% in mass). Additionally, microcalorimetry experiments were performed to estimate the strength of the interaction between PEG and the DNA ligands. Our results suggest that PEG, depending on the system under study, may act as an "inert polymer" with no enthalpic contribution in some processes but, on the other hand, it may as well be an active (non-neutral) osmolyte in the context of modulating the activity of the reactants and products involved in DNA-ligand interactions.

Adição de óleo de palma na dieta sobre a lipidemia e a qualidade do sêmen de bubalinos (Bubalus bubalis) / Addition of palm oil to diet on serum lipid profile and seminal quality of buffaloes (Bubalus bubalis)

O estudo visou investigar o efeito da suplementação dietética com óleo de palma sobre o perfil sérico lipídico e as características seminais de touros bubalinos (Bubalus bubalis), bem como suas correlações. Doze touros foram alocados no Grupo CONT (n=5) ou Grupo ÓLEO (n=7) e receberam alimentação isoproteica composta por silagem de milho e concentrado (milho triturado, farelo de trigo e ureia), com proporção volumoso/concentrado de 50%, durante 130 dias (Períodos 1 e 2). Diferencialmente, óleo de palma foi adicionado ao concentrado (2% MS) do Grupo ÓLEO. Sêmen e sangue foram colhidos quinzenalmente. Houve aumento nas concentrações séricas de colesterol, HDL e lipídios totais nos animais do Grupo ÓLEO, enquanto o período influenciou nas taxas de colesterol, LDL, lipídios totais e VLDL (P<0,05). Houve decréscimo no turbilhonamento e discreta redução na integridade de membranas nos animais do Grupo ÓLEO (P<0,05). Correlações significativas foram encontradas entre triglicerídeos e defeitos menores (r=-0,412; P=0,006), LDL e defeitos totais (r=-0,333; P=0,030), lipídios totais e defeitos menores (r=-0.366; P= 0,017), lipídios totais e defeitos totais (r=-0.309; P= 0,046), e VLDL e viabilidade espermática (r=0,381; P=0,012), apontando efeitos positivos da maior lipidemia na morfologia e na viabilidade espermática. O uso de óleo de palma alterou o perfil lipídico sérico, mas não favoreceu per se os parâmetros seminais relacionados com o potencial de fertilidade dos touros.(AU)

The study aimed to investigate the effect of dietary supplementation with palm oil on serum lipid profile, the seminal characteristics of buffaloes (Bubalus bubalis) and the correlations between them. Twelve bulls were divided into Group CONT (n = 5) or Group OIL (n = 7). Animals in both groups were fed with isoproteic diets composed of corn silage and concentrated (ground corn, wheat bran and urea), with 50% forage/concentrate ratio, during 130 days (Periods 1 and 2). Palm oil (2%DM) was added to diet offered to Group OIL. Semen and blood samples were collected every two weeks. Increased serum levels of cholesterol, HDL and total lipids were observed for Group OIL, while the period influenced on cholesterol, LDL, total lipids and VLDL (P<0,05). Treatment decreased gross motility (P<0.05) and slightly reduced sperm membrane integrity (P<0.05) in animals of Group OIL. There were significant correlations between triglycerides and minor defects (r=-0.412, P=0.006), LDL and total defects (r=-0.333, P=0.030), total lipids and minor defects (r=-0.366, P=0.017), VLDL and spermatic viability (r=0.381, P=0.012), and total lipids and totals defects (r=-0.309, P=0.046), highlighting positive effects of higher lipid serum levels on sperm morphology and viability. The incorporation of palm oil in the diet altered the serum lipid profile, but it showed no positive effect on seminal parameters related to the potential of bull fertility.(AU)

Adição de óleo de palma na dieta sobre a lipidemia e a qualidade do sêmen de bubalinos (Bubalus bubalis) / Addition of palm oil to diet on serum lipid profile and seminal quality of buffaloes (Bubalus bubalis)

O estudo visou investigar o efeito da suplementação dietética com óleo de palma sobre o perfil sérico lipídico e as características seminais de touros bubalinos (Bubalus bubalis), bem como suas correlações. Doze touros foram alocados no Grupo CONT (n=5) ou Grupo ÓLEO (n=7) e receberam alimentação isoproteica composta por silagem de milho e concentrado (milho triturado, farelo de trigo e ureia), com proporção volumoso/concentrado de 50%, durante 130 dias (Períodos 1 e 2). Diferencialmente, óleo de palma foi adicionado ao concentrado (2% MS) do Grupo ÓLEO. Sêmen e sangue foram colhidos quinzenalmente. Houve aumento nas concentrações séricas de colesterol, HDL e lipídios totais nos animais do Grupo ÓLEO, enquanto o período influenciou nas taxas de colesterol, LDL, lipídios totais e VLDL (P<0,05). Houve decréscimo no turbilhonamento e discreta redução na integridade de membranas nos animais do Grupo ÓLEO (P<0,05). Correlações significativas foram encontradas entre triglicerídeos e defeitos menores (r=-0,412; P=0,006), LDL e defeitos totais (r=-0,333; P=0,030), lipídios totais e defeitos menores (r=-0.366; P= 0,017), lipídios totais e defeitos totais (r=-0.309; P= 0,046), e VLDL e viabilidade espermática (r=0,381; P=0,012), apontando efeitos positivos da maior lipidemia na morfologia e na viabilidade espermática. O uso de óleo de palma alterou o perfil lipídico sérico, mas não favoreceu per se os parâmetros seminais relacionados com o potencial de fertilidade dos touros.

The study aimed to investigate the effect of dietary supplementation with palm oil on serum lipid profile, the seminal characteristics of buffaloes (Bubalus bubalis) and the correlations between them. Twelve bulls were divided into Group CONT (n = 5) or Group OIL (n = 7). Animals in both groups were fed with isoproteic diets composed of corn silage and concentrated (ground corn, wheat bran and urea), with 50% forage/concentrate ratio, during 130 days (Periods 1 and 2). Palm oil (2%DM) was added to diet offered to Group OIL. Semen and blood samples were collected every two weeks. Increased serum levels of cholesterol, HDL and total lipids were observed for Group OIL, while the period influenced on cholesterol, LDL, total lipids and VLDL (P<0,05). Treatment decreased gross motility (P<0.05) and slightly reduced sperm membrane integrity (P<0.05) in animals of Group OIL. There were significant correlations between triglycerides and minor defects (r=-0.412, P=0.006), LDL and total defects (r=-0.333, P=0.030), total lipids and minor defects (r=-0.366, P=0.017), VLDL and spermatic viability (r=0.381, P=0.012), and total lipids and totals defects (r=-0.309, P=0.046), highlighting positive effects of higher lipid serum levels on sperm morphology and viability. The incorporation of palm oil in the diet altered the serum lipid profile, but it showed no positive effect on seminal parameters related to the potential of bull fertility.

Microcalorimetric study of the adsorption of lactoferrin in supermacroporous continuous cryogel with immobilized Cu(2+) ions.

The adsorption affinity of lactoferrin from whey in monolithic supermacroporous cryogel was analyzed using equilibrium data adsorptive isothermal titration microcalorimetry to measure thermodynamic information governing the process. Isotherm data was obtained at temperatures of 20, 30 and 40°C, pH 6, 7 and 8, and ionic strength of 200, 600 and 1000mmolL(-1) NaCl. The Langmuir model was fitted to equilibrium data. The binding was tighter at higher temperatures. The adsorption of protein was observed as spontaneous in all cases analyzed. The microcalorimetric study indicated that, in most cases examined, the adsorption of the protein in the matrix was entropy and enthalpy favored and entropy driven. Results provide data to enable the improvement of technical processes for the affinity separation of proteins.

Bat assemblages in conservation areas of a metropolitan region in Southeastern Brazil, including an important karst habitat.

Species richness and abundance of bats were studied in four nature reserves, including a karst area which has many potential rocky shelters for bats, such as caves and rock crevices. The reserves were located in the greater Belo Horizonte metropolitan area, one of the most populated regions of Brazil, within the Atlantic Forest, and Cerrado (Brazilian savanna) ecological domains. Bats were sampled using mist-nets and, in the karst area, also by active searches in shelters. A total of 1,599 bats were captured representing 30 species belonging to four families. There was little similarity among the four chiropteran faunas. The greatest species richness was found in the karst area with 22 species recorded whereas richness estimates in the other areas indicated the need for further studies. Two hundred and sixty-five individuals of 14 species were captured from 56 shelters. Most of the shelters were frequently used for diurnal roosts, and all the bats found belonged to the Phyllostomidae, with the exception of Myotis nigricans (Vespertilionidae), Nyctinomops laticaudatus (Molossidae) and Peropteryx macrotis (Emballonuridae). The sanguinivorous Desmodus rotundus was the most common species in the shelters. The results of this study demonstrate the importance of maintaining multiple protected areas to ensure a representative fauna of bats in a region characterized by a vegetation transition zone and with intense economic activity and high environmental impact. This study also demonstrates the importance of rock shelters for maintaining local bat richness and the importance of active searches for bats in their diurnal roosts for a more thorough sampling of the bat fauna at a given locality.

Bat assemblages in conservation areas of a metropolitan region in southeastern Brazil, including an important karst habitat / Assembleias de morcegos em áreas de conservação de uma região metropolitana no sudeste do Brasil, incluindo um importante hábitat cárstico

Species richness and abundance of bats were studied in four nature reserves, including a karst area which has many potential rocky shelters for bats, such as caves and rock crevices. The reserves were located in the greater Belo Horizonte metropolitan area, one of the most populated regions of Brazil, within the Atlantic Forest, and Cerrado (Brazilian savanna) ecological domains. Bats were sampled using mist-nets and, in the karst area, also by active searches in shelters. A total of 1,599 bats were captured representing 30 species belonging to four families. There was little similarity among the four chiropteran faunas. The greatest species richness was found in the karst area with 22 species recorded whereas richness estimates in the other areas indicated the need for further studies. Two hundred and sixty-five individuals of 14 species were captured from 56 shelters. Most of the shelters were frequently used for diurnal roosts, and all the bats found belonged to the Phyllostomidae, with the exception of Myotis nigricans (Vespertilionidae), Nyctinomops laticaudatus (Molossidae) and Peropteryx macrotis (Emballonuridae). The sanguinivorous Desmodus rotundus was the most common species in the shelters. The results of this study demonstrate the importance of maintaining multiple protected areas to ensure a representative fauna of bats in a region characterized by a vegetation transition zone and with intense economic activity and high environmental impact. This study also demonstrates the importance of rock shelters for maintaining local bat richness and the importance of active searches for bats in their diurnal roosts for a more thorough sampling of the bat fauna at a given locality.(AU)

Neste estudo, a riqueza e a abundância de espécies de morcegos foram estudadas em quatro reservas naturais, incluindo uma área cárstica com muitas cavernas e fendas em rochas, que constituem abrigos rochosos potenciais para os morcegos. As reservas encontram-se dentro da região metropolitana de Belo Horizonte-MG, uma das mais povoadas do Brasil, e dentro dos domínios de Floresta Atlântica e do Cerrado. Os morcegos foram amostrados com redes de neblina (mist-nets) e, na área cárstica, também foi realizada busca ativa por morcegos em seus abrigos. Foram capturados 1.599 morcegos de 30 espécies, pertencentes a quatro famílias. Houve pouca similaridade entre as faunas de quirópteros analisadas. A maior riqueza foi encontrada na área cárstica, com 22 espécies registradas com redes de neblina e em seus abrigos; entretanto, estimativas de riqueza indicaram a necessidade de mais estudos nas outras áreas. Do total amostrado, 265 indivíduos de 14 espécies foram capturados em 56 abrigos. A maioria dos abrigos era ocupada no período diurno e de uso frequente; os morcegos presentes eram filostomídeos, exceto Myotis nigricans (Vespertilionidae), Nyctinomops laticaudatus (Molossidae) e Peropteryx macrotis (Emballonuridae). O hematófago Desmodus rotundus foi o mais frequente nos abrigos. O resultado deste estudo indica a importância da manutenção de diferentes áreas para a conservação dos quirópteros em região caracterizada como zona de transição vegetacional e com intensa atividade econômica e impacto ambiental. Este estudo demonstra também a importância dos abrigos rochosos para a riqueza local de espécies e a importância de busca ativa por morcegos em seus abrigos diurnos para uma amostragem mais efetiva da fauna local destes quirópteros.(AU)

Effects of pregnancy and feeding level on carcass and meat quality traits of Nellore cows.

Carcass and meat quality traits of 16 pregnant and 5 non-pregnant cows fed at 1.2 times maintenance and 16 pregnant and 6 non-pregnant fed ad libitum were evaluated. Pregnancy did not affect final body weight (FBW; P=0.0923), cold carcass yield (CCY; P=0.0513), longissimus muscle area (LMA; P=0.8260), rib fat thickness (RFT; P=0.1873) and shear force (WBSF; P=0.9707). A lower FBW (P=0.0028), LMA (P=0.0048) and RFT (P=0.0001) were observed in feed restricted cows. However, no differences were found for CCY (P=0.7243) and WBSF (P=0.0759) among feeding level groups. These data suggests that carcass and meat quality traits are not affected by pregnancy status in Nellore cows. Moreover, although cows experiencing feed restriction did have reduced deposition of subcutaneous fat and lean tissue, there were no major impacts on meat quality traits.

Ceramic primer heat-treatment effect on resin cement/Y-TZP bond strength.

The purpose of this study was to evaluate the effect of different heat-treatment strategies for a ceramic primer on the shear bond strength of a 10-methacryloyloxydecyl-dihydrogen-phosphate (MDP)-based resin cement to a yttrium-stabilized tetragonal zirconia polycrystal (Y-TZP) ceramic. Specimens measuring 4.5 × 3.5 × 4.5 mm(3) were produced from Y-TZP presintered cubes and embedded in polymethyl methacrylate (PMMA). Following finishing, the specimens were cleaned using an ultrasound device and distilled water and randomly divided into 10 experimental groups (n=14) according to the heat treatment of the ceramic primer and aging condition. The strategies used for the experimental groups were: GC (control), without primer; G20, primer application at ambient temperature (20°C); G45, primer application + heat treatment at 45°C; G79, primer application + heat treatment at 79°C; and G100, primer application + heat treatment at 100°C. The specimens from the aging groups were submitted to thermal cycling (6000 cycles, 5°C/55°C, 30 seconds per bath) after 24 hours. A cylinder of MDP-based resin cement (2.4 mm in diameter) was constructed on the ceramic surface of the specimens of each experimental group and stored for 24 hours at 37°C. The specimens were submitted to a shear bond strength test (n=14). Thermal gravimetric analysis was performed on the ceramic primer. The data obtained were statistically analyzed by two-way analysis of variance and the Tukey test (α=0.05). The experimental group G79 without aging (7.23 ± 2.87 MPa) presented a significantly higher mean than the other experimental groups without aging (GC: 2.81 ± 1.5 MPa; G20: 3.38 ± 2.21 MPa; G100: 3.96 ± 1.57 MPa), showing no difference from G45 only (G45: 6 ± 3.63 MPa). All specimens of the aging groups debonded during thermocycling and were considered to present zero bond strength for the statistical analyses. In conclusion, heat treatment of the metal/zirconia primer improved bond strength under the initial condition but did not promote stable bonding under the aging condition.

Leishmanicidal effects of piperine, its derivatives, and analogues on Leishmania amazonensis.

Leishmaniasis is a tropical disease caused by protozoan parasites of the genus Leishmania which affects 12 million people worldwide. The discovery of drugs for the treatment of leishmaniasis is a pressing concern in global health programs. The aim of this study aim was to evaluate the leishmanicidal effect of piperine and its derivatives/analogues on Leishmania amazonensis. Our results showed that piperine and phenylamide are active against promastigotes and amastigotes in infected macrophages. Both drugs induced mitochondrial swelling, loose kinetoplast DNA, and led to loss of mitochondrial membrane potential. The promastigote cell cycle was also affected with an increase in the G1 phase cells and a decrease in the S-phase cells, respectively, after piperine and phenylamide treatment. Lipid analysis of promastigotes showed that piperine reduced triglyceride, diacylglycerol, and monoacylglycerol contents, whereas phenylamide only reduced diacylglycerol levels. Both drugs were deemed non toxic to macrophages at 50 µM as assessed by XTT (sodium 2,3,-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)-carbonyl]-2H-tetrazolium inner salt), Trypan blue exclusion, and phagocytosis assays, whereas low toxicity was noted at concentrations higher than 150 µM. None of the drugs induced nitric oxide (NO) production. By contrast, piperine reduced NO production in activated macrophages. The isobologram analysis showed that piperine and phenylamide acted synergistically on the parasites suggesting that they affect different target mechanisms. These results indicate that piperine and its phenylamide analogue are candidates for development of drugs for cutaneous leishmaniasis treatment.

Cloning of the transmembrane glycoproteins G and F from a Brazilian isolate of bovine respiratory syncytial virus in a prokaryotic system / Clonagem das glicoproteínas transmembrana G e F de um isolado brasileiro do vírus respiratório sincicial bovino em sistema procarioto

The aim of this work was the cloning of those transmembrane glycoproteins G and F from an isolate bovine respiratory syncytial viruses (BRSV) - a Brazilian isolate of BRSV, named BRSV-25-BR in previous studies, in a prokaryotic system to proceed the sequencing of larger genomic fragments. The nucleotide substitutions were confirmed and these clones may also be used in further studies regarding the biological effects of those proteins in vitro and in vivo.(AU)

O objetivo deste trabalho foi a clonagem das glicoproteínas transmembrana G e F de um isolado de vírus respiratório sincicial bovino (BRSV) - um isolado brasileiro denominado BRSV-25-BR- que já demonstrou possuir mutações em regiões altamente conservadas do gene da proteína G - em sistema procariótico, com o intuito de sequenciar fragmentos genômicos maiores. As substituições de nucleotídeos foram confirmadas e tais clones podem ser utilizados em futuros estudos sobre os efeitos biológicos destas proteínas tanto in vitro como in vivo.(AU)

Clonagem das glicoproteínas transmembrana G e F de um isolado brasileiro do vírus respiratório sincicial bovino em sistema procarioto / Cloning of the transmembrane glycoproteins G and F from a Brazilian isolate of bovine respiratory syncytial virus in a prokaryotic system

The aim of this work was the cloning of those transmembrane glycoproteins G and F from an isolate bovine respiratory syncytial viruses (BRSV) - a Brazilian isolate of BRSV, named BRSV-25-BR in previous studies, in a prokaryotic system to proceed the sequencing of larger genomic fragments. The nucleotide substitutions were confirmed and these clones may also be used in further studies regarding the biological effects of those proteins in vitro and in vivo.

O objetivo deste trabalho foi a clonagem das glicoproteínas transmembrana G e F de um isolado de vírus respiratório sincicial bovino (BRSV) - um isolado brasileiro denominado BRSV-25-BR- que já demonstrou possuir mutações em regiões altamente conservadas do gene da proteína G - em sistema procariótico, com o intuito de sequenciar fragmentos genômicos maiores. As substituições de nucleotídeos foram confirmadas e tais clones podem ser utilizados em futuros estudos sobre os efeitos biológicos destas proteínas tanto in vitro como in vivo.

Genetic diversity of avian infectious bronchitis virus isolated from domestic chicken flocks and coronaviruses from feral pigeons in Brazil between 2003 and 2009.

To detect the presence of infectious bronchitis virus or avian coronavirus, a nested reverse transcriptase PCR (RT-PCR) method was developed with the aim of amplifying a fragment of 530 bases, comprising the gene coding S1 protein. In the first step, all samples were submitted to RNA extraction, RT-PCR, and nested PCR. Next, only the positive nested-PCR samples were propagated in specific-pathogen-free (SPF) embryonated chicken eggs for virus isolation. Positive samples were then sequenced and analyzed using a molecular phylogeny approach. Tracheal swab samples were collected from 23 different domestic chickens distributed in three regions of Brazil, in the period between 2003 and 2009. Also analyzed were six swab samples (tracheal and cloacal) from asymptomatic pigeons (Columba livia), caught in an urbanized region in southeastern Brazil. The study revealed two major phylogenetic groups: one clustered with the Massachusetts vaccine serotype and another joined with the D207 strain. Interestingly, samples grouped with the Connecticut and Arkansas serotypes were also found. Pigeon isolates clustered with the Massachusetts serotype showed significant similarity (close to 100%) to those obtained from chickens. Only one pigeon isolate was seen to be grouped with the Connecticut serotype, and no correlation was observed between sample grouping and region origin. Understanding the diversity of genotypes and eco-epizootiology of the disease in different environments is expected to be helpful for vaccine production aimed at the main circulating variants. In this respect, one could also expect benefits in the management of other bird species that may act as avian coronavirus reservoirs.