RESUMO
A study was conducted on 21 pig herds using one-site production system in the southeast region of Brazil to assess the relationships among serological results for primary pathogens involved in respiratory diseases (Actinobacillus pleuropneumoniae, App; Mycoplasma hyopneumoniae, Mhyo; and swine influenza virus, SIV), cough index, pneumonia index, pleuritis and herd characteristics. The prevalence of antibodies against Mhyo and SIV increased throughout the raising phases, with the highest prevalence in slaughtered pigs (> 40%), while pigs in 65% (14/21) of nurseries demonstrated marked seroprevalence of App that decreased until the day of slaughter. Pleuritis and pulmonary consolidations were recorded in 9.0 and 72.4%, respectively, of the 908 evaluated lungs. Histopathological analysis of the lung lesions revealed suppurative bronchopneumonia in almost half of the lungs (48.9%). Regression analyses were conducted to identify risk factors associated with the cough index; pleuritis; pulmonary consolidation; and App, Mhyo and SIV serological results. All-in-all-out management in nursery buildings reduced the seroprevalence of Mhyo in herds. App seroprevalence was associated with pleuritis, and the presence of cough episodes in growing pigs was associated with SIV seropositivity in nursery pigs.
Assuntos
Infecções por Actinobacillus/veterinária , Infecções por Orthomyxoviridae/veterinária , Pleurisia/veterinária , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/microbiologia , Infecções por Actinobacillus/epidemiologia , Infecções por Actinobacillus/patologia , Actinobacillus pleuropneumoniae/isolamento & purificação , Criação de Animais Domésticos , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antivirais/sangue , Brasil/epidemiologia , Tosse/microbiologia , Tosse/veterinária , Estudos Transversais , Fazendas , Modelos Logísticos , Pulmão/patologia , Mycoplasma hyopneumoniae/isolamento & purificação , Infecções por Orthomyxoviridae/epidemiologia , Pleurisia/epidemiologia , Pleurisia/microbiologia , Pleurisia/patologia , Pneumonia Suína Micoplasmática/epidemiologia , Análise de Regressão , Fatores de Risco , Estudos Soroepidemiológicos , Suínos , Doenças dos Suínos/patologia , Doenças dos Suínos/prevenção & controleRESUMO
Pyricularia oryzae is a species complex that causes blast disease on more than 50 species of poaceous plants. Pyricularia oryzae has a worldwide distribution as a rice pathogen and in the last 30 years emerged as an important wheat pathogen in southern Brazil. We conducted phylogenetic analyses using 10 housekeeping loci for 128 isolates of P. oryzae sampled from sympatric populations of wheat, rice, and grasses growing in or near wheat fields. Phylogenetic analyses grouped the isolates into three major clades. Clade 1 comprised isolates associated only with rice and corresponds to the previously described rice blast pathogen P. oryzae pathotype Oryza (PoO). Clade 2 comprised isolates associated almost exclusively with wheat and corresponds to the previously described wheat blast pathogen P. oryzae pathotype Triticum (PoT). Clade 3 contained isolates obtained from wheat as well as other Poaceae hosts. We found that Clade 3 is distinct from P. oryzae and represents a new species, Pyricularia graminis-tritici (Pgt). No morphological differences were observed among these species, but a distinctive pathogenicity spectrum was observed. Pgt and PoT were pathogenic and highly aggressive on Triticum aestivum (wheat), Hordeum vulgare (barley), Urochloa brizantha (signal grass), and Avena sativa (oats). PoO was highly virulent on the original rice host (Oryza sativa), and also on wheat, barley, and oats, but not on signal grass. We conclude that blast disease on wheat and its associated Poaceae hosts in Brazil is caused by multiple Pyricularia species. Pyricularia graminis-tritici was recently found causing wheat blast in Bangladesh. This indicates that P. graminis-tritici represents a serious threat to wheat cultivation globally.
RESUMO
Porcine enteropathogenic Escherichia coli (PEPEC) produce an outer membrane protein (intimin) called Paa (porcine attaching and effacing-associated), which is involved in the pathogenesis of E. coli in piglets with diarrhea. The paa gene of a PEPEC strain isolated in Paraná, Brazil, was amplified by polymerase chain reaction, sequenced, and cloned into the pTrcHisTOPO2 vector. The deduced amino acid sequence encoded by the paa gene of PEPEC from Paraná, Brazil, showed 99% homology to the sequences from other PEPEC strains. In this study, the overexpression of recombinant Paa (rPaa) using alternative induction strategies was attempted. The auto-induction protocol showed excellent results for rPaa protein production with 0.4% (w/v) lactose. The rPaa protein is insoluble and was purified with Triton X-100 wash as a total antigen. This method produced a relatively high yield of rPaa. rPaa was recognized by serum from pigs immunized with the PEPEC strain. These results suggest that rPaa could be included in the development of a vaccine against swine colibacillosis.
Assuntos
Escherichia coli Enteropatogênica/genética , Infecções por Escherichia coli/veterinária , Proteínas de Escherichia coli/genética , Animais , Clonagem Molecular , Escherichia coli Enteropatogênica/isolamento & purificação , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/prevenção & controle , Proteínas de Escherichia coli/biossíntese , Expressão Gênica , Sus scrofa/microbiologia , Suínos , Doenças dos Suínos/microbiologia , Doenças dos Suínos/prevenção & controle , Ativação TranscricionalRESUMO
BACKGROUND: Gait speed is considered a predictor of adverse health outcomes and functional decline in the elderly. This decline is also identified in respiratory muscles. OBJECTIVE: To assess the impact of gait speed in maximal inspiratory pressure, maximal expiratory pressure, handgrip strength, and the different types of frailty syndrome in community-dwelling elderly people. DESIGN: Cross-sectional study. PARTICIPANTS: Women (aged ≥ 65 years) were classified into different frailty phenotypes (n = 106). MEASUREMENTS: Gait speed (10 m), handgrip strength (Jamar dynamometer), and maximum inspiratory and expiratory pressures (GerAr manovacuometer, MV-150/300 model) were measured. Linear regression analyses were conducted to determine the influence of gait speed and age on handgrip strength, maximal inspiratory pressure, and maximal expiratory pressure. Logistic regression was performed to assess the influence of gait speed and frailty age (α = 0.05). RESULTS: A total of 106 elderly women participated in the study (73.96 ± 6.91 years). Thirty-two subjects were not frail, 42 were pre-frail, and 32 were frail. Gait speed and age significantly predicted handgrip strength and frailty (p < 0.05). In the multivariate model, gait speed had the greatest contribution, while age lost statistical significance. Regarding maximal inspiratory and maximal expiratory pressures, gait speed and age were significant explanatory variables (p < 0.05). In the multivariate model, gait speed lost statistical significance to predict maximal inspiratory pressure. CONCLUSION: Gait speed was confirmed to be a predictor of some health outcomes, including respiratory muscle function. The results suggest that interventions to increase gait speed may contribute to improve respiratory function and muscle strength, and decrease the risk of frailty among elderly people.
RESUMO
Composition and spatial diversity patterns of retained and discarded catches in the deep-water shrimp (family Aristeidae) trawling fishery off Brazil were assessed by observers on-board commercial operations in 2005 and 2006. These trawls caught 19,440 kg and 180,076 individuals of which 76·0 and 65·2%, respectively, were discarded at sea. Finfishes represented 54% of the numerical catch but were almost fully discarded (98%). Crustaceans represented 40% of the numerical catch and were mostly retained (80%). The scarlet shrimp Aristaeopsis edwardsiana comprised approximately half of the retained catch. The remainder of the retained proportion comprised mainly the red giant shrimp Aristaeomorpha foliacea, the alistado shrimp Aristeus antillensis and small quantities of Argentine hake Merluccius hubbsi and gulf hake Urophycis mystacea. Discards comprised 108 species including 72 fish species, 19 crustaceans and 10 cephalopods. The large-scaled lanternfish Neoscopelus macrolepidotus was dominant in the discards, followed by the benthopelagic fishes Monomitopus agassizii, Synagrops bellus, Dibranchus atlanticus and Gadella imberbis and various macrurid species. This fishery was restricted to a limited bathymetric range (700-800 m), where discrete megafauna assemblages exist and may have been significantly affected.
Assuntos
Biota , Pesqueiros , Animais , Brasil , Cefalópodes , Decápodes , PeixesRESUMO
A metadona é um opioide pouco utilizado na medicina veterinária, e há, ainda, questionamentos quanto ao seu uso. Desse modo, comparou-se o emprego da metadona, administrada pelas vias IM e IV, e avaliaram-se seus efeitos sobre as características cardiorrespiratórias e os tempos de extubação e recuperação em cadelas submetidas à ovariossalpingo-histerectomia. Utilizaram-se 16 cadelas, pré-medicadas com levomepromazina, 0.6mg/kg, IM. Após 20 minutos, a indução anestésica foi feita com propofol, 5mg/kg, IV, e a manutenção com isoflurano. Transcorridos 10 minutos, os animais receberam metadona, 0.3mg/kg, pelas vias IM - grupo GIM - e IV - grupo GIV. As mensurações das frequências cardíaca (FC) e respiratória (FR), pressão arterial média (PAM), temperatura esofágica (TE), pressão parcial de dióxido de carbono (EtCO2) e saturação de oxi-hemoglobina (SpO2) foram registradas antes da administração do opioide (T0), após 20 minutos (T1) e a cada 10 minutos da administração até 60 minutos (T2 a T5). Usou-se a análise de perfil (5%). Com relação à FC, PAM, FR, EtCO2, TE e SpO2, não foram observadas diferenças significativas entre grupos e momentos. Observaram-se tempos de extubação e recuperação maiores no GIV. É possível concluir que a administração IV e a IM da metadona não produzem alterações nas características cardiorrespiratórias em cadelas.
Methadone is a little used opioid in veterinary practice, and there are still questions about its use. The objective of this study was to evaluate the effects of intramuscular (IM) or intravenous (IV) administration of methadone on cardiopulmonary parameters and times of extubation and recovery in female dogs submitted to ovariohysterectomy. Sixteen adult female dogs were used and premedicated with levomepromazine (0.6mg/kg, IM). After 20 minutes, propofol (5mg/kg, IV) was used for induction and anesthesia was maintained with isoflurane. After 10 minutes, methadone at 0.3mg/kg was administered intravenously in IVG and intramuscularly in IMG. The measurement of heart (HR) and respiratory rates (RR), median arterial pressure (MAP), esophagic temperature (ET) and concentration of end-tidal carbon dioxide (PE'CO2) was performed immediately before the administration of the opioid (T0), after 20 minutes (T1) and then at 10-minute intervals (T2, T3, T4 and T5). The statistical analysis used was profile (5%). HR, APM, RR, PE'CO2, BT and SpO2 did not differ significantly among times or between groups at any time. Times of extubation and recovery were higher in IVG. It is possible to conclude that IV and IM administration of methadone did not produce changes in the cardiorespiratory parameters of that specie.
Assuntos
Animais , Cães , Analgésicos/análise , Anestesia , Histerectomia , Injeções Intramusculares , Injeções Intravenosas , Cães/classificaçãoRESUMO
The latex of Calotropis procera is a rich source of proteolytic activity. This latex is known to contain two distinct cysteine peptidases: procerain and procerain B. In this study, new cysteine peptidases were purified from C. procera latex. The enzymes were purified by two sequential ion-exchange chromatography steps (CM-Sepharose plus Resource S(®)) at pH 5.0 and 6.0. The purified enzymes had molecular mass spectra corresponding to CpCP-1=26,213, CpCP-2=26,133 and CpCP-3=25,086 Da. These enzymes exhibited discrete differences in terms of enzymatic activity at a broad range of pH and temperature conditions and contained identical N-terminal amino acid sequences. In these respects, these three new proteins are distinct from those previously studied (procerain and procerain B). Circular dichroism analysis revealed that the new peptidases contain extensive secondary structures, α(15-20%) and ß(26-30%), that were stabilized by disulfide bonds. The purified enzymes exhibited plasma-clotting activity mediated by a thrombin-like mechanism. The set of results suggest the three isolated polypeptides correspond to different post-translationally processed forms of the same protein.
Assuntos
Calotropis/enzimologia , Coagulantes/química , Cisteína Endopeptidases/química , Látex/química , Proteínas de Plantas/química , Sequência de Aminoácidos , Coagulação Sanguínea , Cromatografia por Troca Iônica , Coagulantes/isolamento & purificação , Coagulantes/farmacologia , Cisteína Endopeptidases/isolamento & purificação , Cisteína Endopeptidases/farmacologia , Humanos , Concentração de Íons de Hidrogênio , Hidrólise , Dados de Sequência Molecular , Peso Molecular , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/farmacologia , Estrutura Secundária de Proteína , Proteólise , Tempo de Protrombina , Análise de Sequência de Proteína , Homologia de Sequência de AminoácidosRESUMO
BACKGROUND AND OBJECTIVE: Gingivitis is a disease that is characterized by inflammation of the gingival tissue, which can progress to periodontitis and tooth loss. Although many studies have attempted to identify salivary proteins that are associated with the disease, this is the first study to use a proteomic approach to analyze and compare the proteomic profile of whole saliva from gingivitis patients and healthy controls. MATERIAL AND METHOD: To analyze the saliva proteome, two-dimensional gel electrophoresis and liquid chromatography were used, followed by mass spectrometry. RESULTS: The analyses showed that gingival inflammation was associated with increased amounts of blood proteins (serum albumin and hemoglobin), immunoglobulin peptides and keratins. In the control group, salivary cystatins, which were detected using capillary Liquid Chromatography on line to electrospray ionization Quadrupole Time-of-flight mass spectrometry, appeared to be more abundant. CONCLUSION: This approach provides novel insight into profiles of the salivary proteome during gingival inflammation, which may contribute to improvements in diagnosis.
Assuntos
Gengivite/metabolismo , Proteoma , Proteínas e Peptídeos Salivares/análise , Adulto , Cromatografia Líquida , Bases de Dados de Proteínas , Eletroforese em Gel Bidimensional , Feminino , Perfilação da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Empregou-se a associação midazolam e detomidina para indução de anestesia com cetamina em 16 potros, machos e fêmeas, entre três e seis meses de idade, distribuídos aleatória e equitativamente em dois grupos (GI e GII). A todos os animais foram administrados midazolam, via intramuscular, na dose de 0,2mg/kg, e após 15 minutos, detomidina, via intravenosa, na dose de 0,02mg/kg. Os animais do GII receberam cetamina pela via intravenosa, dose 2,0mg/kg, três minutos após a administração de detomidina. Quinze minutos após o midazolam, ocorreram sedação e ligeira ataxia, e dois minutos após a administração da detomidina, decúbito lateral em todos os potros, com miorrelaxamento e presença dos reflexos de deglutição e miorrelaxamento, anal e oculo-palpebral. A associação midazolam/detomidina e cetamina provocou ausência dos reflexos de deglutição. Para todos os animais, o tempo de recuperação foi de 45-60 minutos, e temperatura retal e frequência respiratória permaneceram estáveis. Ocorreram bradicardia, bloqueio atrioventricular de segundo grau e aumento das pressões arteriais sistólica, diastólica e média após dois minutos da administração da detomidina. A associação midazolam/detomidina e cetamina demonstrou ser um método eficiente e seguro para a anestesia de potros hígidos.
A combination of midazolam, 0.2mg/kg body weight given via intramuscular, and detomidine, 0.02mg/kg body weight given via intravenous (IV), was evaluated as a method for induction of anesthesia with ketamine, 2.0mg/kg body weight given via IV in foals. Sixteen male and female foals aging from three to six-month old were distributed into two groups. Both groups were first injected with midazolam and with detomidine 15 minutes later. Three minutes later, ketamine was injected in the foals. Sedation and light ataxia were observed 15 minutes after midazolam administration. Bradycardia, atrioventricular block, increased blood pressure, lateral recumbency, and muscle relaxation were observed two minutes after detomidine. At that time, the oculopalpebral, anal, and cogh reflexes were present. The association of midazolam/detomidine and ketamine caused absence of the swallow reflexes. The recovery time was from 45 to 60 minutes for animals from GI and GII groups, respectively. Respiratory rate and body temperature remained unchanged. The combination of midazolam/detomidine and ketamine is a safe, good, and efficient method for anesthesia in healthy foals.
Assuntos
Animais , Analgésicos/administração & dosagem , Anestésicos/administração & dosagem , Combinação de Medicamentos , Midazolam/administração & dosagem , CavalosRESUMO
Foram realizados estudos empregando-se analgésicos por via epidural e subcutânea em cadelas de diferentes raças e idades, submetidas à castração mediante celiotomia. Vinte animais foram tranquilizados e anestesiados com tiletamina-zolazepam, e aleatoriamente distribuídos em quatro grupos (n=5), de acordo com o fármaco e a via de administração. Os do grupo morfina (GM) foram submetidos à anestesia epidural no espaço lombossacro, com morfina (0,1mg/kg) associada ao cloreto de sódio a 0,9 por cento. Aos do grupo xilazina (GX), foram administrados xilazina (0,2mg/kg) e cloreto de sódio a 0,9 por cento. Os do grupo meloxicam (GME) receberam 0,2mg/kg do anti-inflamatório meloxicam associado ao cloreto de sódio a 0,9 por cento, injetado pela via subcutânea. Os do grupo-controle (CG) receberam apenas cloreto de sódio a 0,9 por cento. O volume final para as injeções epidurais foi padronizado para 0,3mL/kg. A mensuração inicial da concentração de cortisol plasmático, do ritmo cardíaco, da frequência respiratória e os parâmetros comportamentais foram registrados imediatamente antes do procedimento cirúrgico (M1). Registros adicionais foram apresentados às 2, 6, 12 e 24 horas após o procedimento cirúrgico (M2, M3, M4 e M5, respectivamente). As variáveis comportamentais foram avaliadas por meio de sinais clínicos e seus respectivos escores. Em GX foram observadas depressão respiratória, bradicardia e concentração de cortisol mais alta do que o registrado no GM. A analgesia obtida pelo meloxicam foi considerada ineficiente. É possível concluir que a morfina, via epidural, promoveu menor incidência de efeitos colaterais e melhor analgesia e bem-estar animal.
The use of analgesics by epidural and subcutaneous way in bitches submitted to surgical sterilization by laparotomy was evaluated. Twenty females dogs of different ages and breeds were sedated and anesthetized with a combination of tiletamine-zolazepam and randomly distributed into four experimental groups of five animals each. Through the epidural space, the animals of each group received 0.1mg/kg of morphine (MG) or 0.2mg/kg of xylazine (XG); while the control group (CG) received a 0.9 percent solution of chloride sodium. The final volume stabilished for lumbosacral epidural injections was 0.3mL/kg. The dogs of meloxicam group (MEG) subcutaneously received 0.2mg/kg of the drug. The initial measurement of plasmatic cortisol concentration, heart and respiratory rates, and behaviour parameters were evaluated as soon as the surgical procedure had finished (M1). These evaluations were repeated after 2, 6, 12, and 24 hours of the post-operative period (M2, M3, M4, and M5 respectively). The behaviour parameters were evaluated by clinical signs and respective scores. Values of cortisol concentration were higher in the xylazine group than the ones registered in the morphine group. Meloxicam failed to provide pain control. Epidural use of morphine guaranteed fewer side effects and better pain control.
Assuntos
Animais , Feminino , Bem-Estar do Animal , Anti-Inflamatórios , Castração , Cães , Histerectomia/métodos , Morfina/administração & dosagem , Dor , Cuidados Pós-Operatórios , Xilazina/administração & dosagemRESUMO
Avaliou-se o efeito de dois fosfatos, tripolifosfato de sódio (TPF) e hexametafosfato de sódio (HMF), incorporados à ração seca sob diferentes formas, como cobertura do grânulo da ração e no interior da massa da ração, na prevenção do desenvolvimento de cálculo dentário em 25 cães. O delineamento experimental utilizado foi inteiramente ao acaso, com cinco tratamentos e cinco repetições. Os tratamentos experimentais foram: 1) ração seca; 2) ração seca com TPF incorporado na cobertura do grânulo da ração; 3) ração seca com TPF incorporado na massa da ração; 4) ração seca com HMF incorporado na cobertura do grânulo da ração; 5) ração seca com HMF incorporado na massa da ração. Os animais receberam as dietas por um período experimental de 90 dias e, após esse período, foram submetidos à avaliação da área de cálculo dentário formado. A inclusão do HMF na ração seca, tanto na cobertura dos grânulos como no interior da massa, e do TPF, como cobertura dos grânulos, reduziu o acúmulo de cálculo dentário em cães, comparada à dieta sem adição de fosfatos. O HMF foi o fosfato mais eficiente, ao reduzir o acúmulo de cálculo dentário em até 47 por cento. As formas de inclusão do HMF na ração não influenciaram o acúmulo de cálculo dentário. Houve redução do desenvolvimento de cálculo dentário pela incorporação do TPF como cobertura do grânulo da ração, comparada à inclusão deste fosfato no interior da massa da ração. Conclui-se que os fosfatos incorporados na ração seca reduzem o acúmulo de cálculo dentário em cães.
The effect of dry food treated with sodium tripolyphosphate (TPP) or sodium hexametaphosphate (HMP), as kibble coated or added into the interior of the kibble on the accumulation of dental calculus in 25 dogs was evaluated. The study used a completely randomized design with five treatments and five repetitions. The experimental treatments were: 1) dry food; 2) dry food coated with TPP; 3) dry food with TPP incorporated into the interior of the kibble; 4) dry food coated with HMP; 5) dry food with HMP incorporated into the interior of the kibble. The animals received the diets for a 90-day experimental period, and then, the teeth were clinically examined for the presence of calculus. The inclusion of HMP in the dry food, as kibble coated or added into the interior of the kibble, and the inclusion of TPP as kibble coated reduced the accumulation of dental calculus compared to the control group without anti-calculus agents. HMP was the most efficient phosphate, reducing the accumulation of dental calculus in 47 percent. The forms of HMP inclusion in the dry food exerted no significant effect upon calculus formation. However, there was a reduction of calculus accumulation when TPF was incorporated as coating of the kibble, compared to the inclusion of this phosphate into the interior of the kibble. It was concluded that phosphates incorporated in the dry food reduce the accumulation of dental calculus in dogs.
Assuntos
Animais , Masculino , Feminino , Ração Animal , Cálculos Dentários/prevenção & controle , Cálculos Dentários/veterinária , Cães , Fosfatos/administração & dosagem , Fosfatos/metabolismo , Fosfatos/uso terapêuticoRESUMO
The main objective of this study was to evaluate the clinical differences and the pattern and extent of organ damage in late-onset systemic lupus erythematosus (SLE). A nested case-control study was performed from patients with SLE followed in the Rheumatology Unit of the State University of Campinas between 1974 and 2005. Patients who developed SLE after the age of 49 were considered late-onset SLE. SLE patients with age <49 years, matched for sex, ethnicity, disease duration and organ damage at study entry were randomly chosen to compose the control group. Baseline and cumulative clinical manifestations, laboratory data, SLE disease activity index (SLEDAI), Systemic Lupus International Collaborating Clinics/American College of Rheumatology-damage index (SDI) and mortality were compared between groups. At diagnosis and follow-up, late-onset group had lower SLEDAI scores when compared with younger age onset. Clinically, they presented less frequently arthritis (P = 0.0002) and malar rash (P = 0.02) and more frequently Raynaud's phenomenon (P = 0.002) and arterial hypertension (P = 0.02) when compared with young onset at diagnosis. Late-onset SLE received lower total corticosteroid dose (P < 0.001) and less frequently cyclophosphamide (P = 0.01). During the study period, late-onset SLE had always lower SLEDAI scores (P = 0.001). At study endpoint, late-onset SLE patients had significantly higher SDI scores (P = 0.001) and a higher mortality rate when compared with younger onset group (P < 0.01). In conclusion, late-onset SLE is milder on presentation and during course of disease, but patients have more organ damage and a higher rate of mortality than young onset SLE. Patients with late onset should be followed with close monitoring and early identification of complications is mandatory in this subgroup of patients with SLE.
Assuntos
Lúpus Eritematoso Sistêmico/diagnóstico , Idade de Início , Idoso , Estudos de Casos e Controles , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Fatores de TempoRESUMO
Avaliou-se a utilização de coadjuvantes na diminuição da placa bacteriana e formação do cálculo dentário em 16 cães. O delineamento experimental foi casualizado com quatro tratamentos (T) e quatro repetições. Os tratamentos foram: T1- controle, T2- coadjuvante com ação mecânica, T3- coadjuvante com tripolifosfato de sódio e T4- coadjuvante com hexametafosfato de sódio. Após sete dias de adaptação, no dia anterior ao início dos tratamentos, os animais foram submetidos à remoção de cálculo dentário. O experimento teve duração de 21 dias e ao final realizaram-se as medições das placas bacterianas formadas com o uso de marcadores (fucsina). O coadjuvante somente com ação mecânica não foi efetivo em retardar o aparecimento da placa bacteriana. Os coadjuvantes com polifosfatos apresentaram uma ação efetiva e significativa na diminuição da formação do calculo dentário
The use of coadjutants in the decrease of the bacterial plate and formation of the dental calculus was evaluated in 16 dogs. The experimental design was randomized with four treatments (T) and four repetitions. The treatments were the following: 1- control; 2- coadjutant 1 (one) with mechanical action; 3- coadjutant 2 (two) with tripolyphosphate of sodium and 4- coadjutant 3 (three) with hexametaphosphate of sodium. After seven days of adaptation, on the day previous to the beginning of the treatments, the animals were submitted to the removal of dental calculus and after twenty-one days of treatment a measurement of the bacterial plaque formed through the use of markers (fucsin). The coadjutant only with mechanical action was not effective in delaying the appearance of the bacterial plaque. The coadjutant containing polyphosphates presented an effective and significant action decreasing the formation of the dental calculus
Assuntos
Animais , Masculino , Feminino , Cães , Cálculos Dentários/prevenção & controle , Cálculos Dentários/veterinária , Cães , Coadjuvantes de Tecnologia Alimentar , Higiene Bucal/métodos , Placa Dentária/prevenção & controle , Placa Dentária/veterinária , Polifosfatos/administração & dosagem , Ração Animal/toxicidadeRESUMO
This article reviews the origin and evolution of high throughput screening (HTS) through the experience of an individual pharmaceutical company, revealing some of the mysteries of the early stages of drug discovery to the wider pharmacology audience. HTS in this company (Pfizer, Groton, USA) had its origin in natural products screening in 1986, by substituting fermentation broths with dimethyl sulphoxide solutions of synthetic compounds, using 96-well plates and reduced assay volumes of 50-100 microl. A nominal 30 mM source compound concentration provided high microM assay concentrations. Starting at 800 compounds each week, the process reached a steady state of 7200 compounds per week by 1989. Screening in the Applied Biotechnology and Screening Group was centralized with screens operating in lock-step to maximize efficiency. Initial screens were full files run in triplicate. Autoradiography and image analysis were introduced for (125)I receptor ligand screens. Reverse transcriptase (RT) coupled with quantitative PCR and multiplexing addressed several targets in a single assay. By 1992 HTS produced 'hits' as starting matter for approximately 40% of the Discovery portfolio. In 1995, the HTS methodology was expanded to include ADMET targets. ADME targets required each compound to be physically detected leading to the development of automated high throughput LC-MS. In 1996, 90 compounds/week were screened in microsomal, protein binding and serum stability assays. Subsequently, the mutagenic Ames assay was adapted to a 96-well plate liquid assay and novel algorithms permitted automated image analysis of the micronucleus assay. By 1999 ADME HTS was fully integrated into the discovery cycle.
Assuntos
Desenho de Fármacos , Avaliação Pré-Clínica de Medicamentos/história , Indústria Farmacêutica/história , Farmacologia/história , Testes de Toxicidade/história , Animais , Linhagem Celular , Sistema Livre de Células , Bases de Dados como Assunto/história , Difusão de Inovações , Avaliação Pré-Clínica de Medicamentos/tendências , Indústria Farmacêutica/métodos , Indústria Farmacêutica/tendências , Técnicas Genéticas , História do Século XX , História do Século XXI , Humanos , Ligantes , Análise em Microsséries/história , Microquímica/história , Estrutura Molecular , Farmacocinética , Farmacologia/métodos , Farmacologia/tendências , Conformação Proteica , Reprodutibilidade dos Testes , Relação Estrutura-Atividade , Testes de Toxicidade/tendências , Estados UnidosRESUMO
O crescimento da tuberculose (TB) pulmonar é um grande problema de Saúde Pública no Brasil. A adesão do paciente ao uso de medicamentos é fundamental, sendo uma das condições essenciais para o êxito do tratamento. A longa duração do tratamento e utilização de medicamentos que usualmente proporcionam reações adversas intensas acabam por comprometer esta adesão. O objetivo deste estudo foi implantar a Metodologia Dáder de Seguimento Farmacoterapêutico aos pacientes portadores de TB pulmonar no Ambulatório de Tisiologiado Hospital e Maternidade Celso Pierro, em Campinas, SP (Brasil). Para tanto foram selecionados sete pacientes de acordo com os critérios de inclusão e exclusão. Através das entrevistas agendadas e o seguimento farmacoterapêutico proposto por esta metodologia, verificou-se que seis pacientes tiveram alta dentro do esquema terapêutico I proposto pelo programa nacional de controle da tuberculose (PNCT); adicionalmente, estes pacientes foram os que tiveram bons níveis de cumprimento da farmacoterapia. Segundo ametodologia, detectou-se dois tipos de problemas relacionados a medicamentos (PRM), referentes à segurança: o PRM6 foi o mais freqüente (seis pacientes), entretanto, um dos pacientes manifestou o PRM5 associado ao PRM6. O estudo realizado com os pacientes que participaram do seguimento farmacoterapêutico mostrou que a Metodologia Dáder é aplicável e eficiente na identificação de PRM, em pacientes portadores deTB e que o farmacêutico no desenvolvimento da Atenção Farmacêutica é um profissional que compõe a equipe de saúde, sendo importante para o sucesso do tratamento farmacológico e não farmacológico.
Assuntos
Humanos , Masculino , Feminino , Adulto , Adesão à Medicação , Antituberculosos/uso terapêutico , Assistência Farmacêutica , Tuberculose Pulmonar/tratamento farmacológico , SeguimentosRESUMO
An intensity-referenced temperature-independent curvature-measurement technique that uses a smart composite that comprises two chirped fiber Bragg gratings is demonstrated. The two gratings are embedded on opposite sides of the composite laminate and act simultaneously as curvature sensors and as wavelength discriminators, enabling a temperature-independent intensity-based scheme to measure radius of curvature. Also, the system's performance is independent of arbitrary power losses that are induced in the lead fibers to the sensing head. It is demonstrated that the measurement range depends on the relative positions of the chirped fiber Bragg gratings and on their spectral bandwidths. By using two chirped fiber Bragg gratings with bandwidths W1 = 2.8 nm and W2 = 3.7 nm and with central wavelengths at lambda 01 = 1560.3 nm and lambda 02 = 1563.7 nm, we obtained a resolution of 1.6 mm/square root of Hz for the measurement of the radius of curvature (approximately R = 350 mm) over the measurement range 190 mm < R < infinity.
RESUMO
Dipeptidyl peptidase IV (DPP-IV; CD26) (EC 3.4.14.5) is a membrane-anchored ectoenzyme with N-terminal exopeptidase activity that preferentially cleaves X-Pro-dipeptides. It can also be spontaneously released to act in the extracellular environment or associated with the extracellular matrix. Many hematopoietic cytokines and chemokines contain DPP-IV-susceptible N-terminal sequences. We monitored DPP-IV expression and activity in murine bone marrow and liver stroma cells which sustain hematopoiesis, myeloid precursors, skin fibroblasts, and myoblasts. RT-PCR analysis showed that all these cells produced mRNA for DPP-IV. Partially purified protein reacted with a commercial antibody to CD26. The K M values for Gly-Pro-p-nitroanilide ranged from 0.43 to 0.98 mM for the membrane-associated enzyme of connective tissue stromas, and from 6.76 to 8.86 mM for the enzyme released from the membrane, corresponding to a ten-fold difference, but only a two-fold difference in K M was found in myoblasts. K M of the released soluble enzyme decreased in the presence of glycosaminoglycans, nonsulfated polysaccharide polymers (0.8-10 æg/ml) or simple sugars (320-350 æg/ml). Purified membrane lipid rafts contained nearly 3/4 of the total cell enzyme activity, whose K M was three-fold decreased as compared to the total cell membrane pool, indicating that, in the hematopoietic environment, DPP-IV activity is essentially located in the lipid rafts. This is compatible with membrane-associated events and direct cell-cell interactions, whilst the long-range activity depending upon soluble enzyme is less probable in view of the low affinity of this form
Assuntos
Animais , Camundongos , Ratos , Células da Medula Óssea , Dipeptidil Peptidase 4 , Células Estromais , Linhagem Celular , Dipeptidil Peptidase 4 , Expressão Gênica , Sistema Hematopoético , Immunoblotting , Fígado , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA MensageiroRESUMO
Dipeptidyl peptidase IV (DPP-IV; CD26) (EC 3.4.14.5) is a membrane-anchored ectoenzyme with N-terminal exopeptidase activity that preferentially cleaves X-Pro-dipeptides. It can also be spontaneously released to act in the extracellular environment or associated with the extracellular matrix. Many hematopoietic cytokines and chemokines contain DPP-IV-susceptible N-terminal sequences. We monitored DPP-IV expression and activity in murine bone marrow and liver stroma cells which sustain hematopoiesis, myeloid precursors, skin fibroblasts, and myoblasts. RT-PCR analysis showed that all these cells produced mRNA for DPP-IV. Partially purified protein reacted with a commercial antibody to CD26. The K M values for Gly-Pro-p-nitroanilide ranged from 0.43 to 0.98 mM for the membrane-associated enzyme of connective tissue stromas, and from 6.76 to 8.86 mM for the enzyme released from the membrane, corresponding to a ten-fold difference, but only a two-fold difference in K M was found in myoblasts. K M of the released soluble enzyme decreased in the presence of glycosaminoglycans, nonsulfated polysaccharide polymers (0.8-10 micro g/ml) or simple sugars (320-350 micro g/ml). Purified membrane lipid rafts contained nearly 3/4 of the total cell enzyme activity, whose K M was three-fold decreased as compared to the total cell membrane pool, indicating that, in the hematopoietic environment, DPP-IV activity is essentially located in the lipid rafts. This is compatible with membrane-associated events and direct cell-cell interactions, whilst the long-range activity depending upon soluble enzyme is less probable in view of the low affinity of this form.
Assuntos
Células da Medula Óssea/enzimologia , Dipeptidil Peptidase 4/fisiologia , Microdomínios da Membrana/enzimologia , Células Estromais/enzimologia , Animais , Linhagem Celular , Dipeptidil Peptidase 4/genética , Expressão Gênica , Sistema Hematopoético/enzimologia , Immunoblotting , Fígado/citologia , Fígado/enzimologia , Camundongos , RNA Mensageiro/biossíntese , Ratos , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
On-line monitoring biomass concentration in mycelial fed-batch cultivations of Streptomyces clavuligerus grown with soluble and partially insoluble complex media, was investigated with an in-situ capacitance probe fitted to an industrial pilot-plant tank. Standard off-line and on-line biomass determinations, including cell dry weight, packed mycelial volume, viscosity, DNA concentration and total CO(2) evolution in the exhaust gases, were performed throughout the experiments and compared to on-line capacitance measurements. Linear relations between capacitance and all other measurements were developed for both media that hold only in defined process phases, depending on the biomass state and the amount of insoluble matter present. For the industrial complex culture media good linear relations were obtained in the fast growth phase between capacitance and DNA concentration and total CO(2) evolution, while in the subsequent transition and stationary phases only with apparent viscosity was a reasonable correlation found. The capacitance probe was shown to be a valuable tool for real-time monitoring biomass concentration in industrial-like cultivation of mycelial streptomycetes.
Assuntos
Ácido Clavulânico/biossíntese , Microbiologia Industrial/instrumentação , Microbiologia Industrial/métodos , Streptomyces/crescimento & desenvolvimento , Streptomyces/metabolismo , Reatores Biológicos , Dióxido de Carbono/metabolismo , Meios de Cultura , DNA Bacteriano/análise , Condutividade Elétrica , Modelos LinearesRESUMO
The XPR2 gene encoding an alkaline extracellular protease (AEP) from Yarrowia lipolytica was cloned, and its complete nucleotide sequence was determined. The amino acid sequence deduced from the nucleotide sequence reveals that the mature AEP consists of 297 amino acids with a relative molecular weight of 30,559. The gene codes for a putative 22-amino-acid prepeptide (signal sequence) followed by an additional 135-amino-acid propeptide containing a possible N-linked glycosylation site and two Lys-Arg peptidase-processing sites. The final Lys-Arg site occurs at the junction with the mature, extracellular form. The mature protease contains two potential glycosylation sites. AEP is a member of the subtilisin family of serine proteases, with 42.6% homology to the fungal proteinase K. The functional promoter is more than 700 base pairs long, allowing for the observed complex regulation of this gene. The 5' and 3' flanking regions of the XPR2 gene have structural features in common with other yeast genes.