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1.
J Virol Methods ; 327: 114948, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38718900

ABSTRACT

Rabies, a fatal zoonotic viral disease affecting mammals, including humans, remains a significant global health concern, particularly in low-income countries. The disease, primarily transmitted through infected animal saliva, prompts urgent diagnosis for timely post-exposure prophylaxis (PEP). The gold standard diagnostic test, direct fluorescent antibody test (dFAT), while sensitive, suffers from limitations such as subjective interpretation and high costs. As a confirmatory technique, the LN34 Pan-Lyssavirus RT-qPCR assay has emerged as a promising tool for universal Lyssavirus detection. This study evaluated its performance using 130 rabies virus isolates representing eleven Brazilian variants and 303 clinical samples from surveillance operations. The LN34 assay demonstrated 100% sensitivity and 98% specificity compared to dFAT. Additionally, it detected all samples, including those missed by dFAT, indicating superior sensitivity. The assay's specificity was confirmed through Sanger nucleotide sequencing, with only a minimal false-positive rate. Comparative analysis revealed higher accuracy and concordance with dFAT than traditional rabies tissue culture infection tests (RTCIT). False-negative RTCIT results were attributed to low viral load or suboptimal sampling. These findings underscore the LN34 assay's utility as a confirmatory technique, enhancing rabies surveillance and control in Brazil. Its widespread adoption could significantly improve diagnostic sensitivity, crucial for effective PEP and public health interventions.


Subject(s)
Rabies virus , Rabies , Real-Time Polymerase Chain Reaction , Sensitivity and Specificity , Rabies/diagnosis , Rabies/veterinary , Rabies/virology , Brazil , Rabies virus/genetics , Rabies virus/isolation & purification , Rabies virus/classification , Humans , Animals , Real-Time Polymerase Chain Reaction/methods , Lyssavirus/genetics , Lyssavirus/isolation & purification , Lyssavirus/classification , RNA, Viral/genetics , Viral Load
2.
J Virol Methods ; 283: 113918, 2020 09.
Article in English | MEDLINE | ID: mdl-32554044

ABSTRACT

The direct-fluorescent antibody test (dFAT) is considered the "gold standard" assay to diagnose rabies. However, it is crucial to develop molecular techniques, such as RT-PCR and RT-qPCR, since many laboratories lack the needed supplies for performing complementary methods (viral isolation, for example). For this purpose, diagnostic techniques must be specific and sensitive to guarantee accuracy. This present investigation aimed to detect rabies virus (RABV) in 126 clinically suspected cattle in Brazil using different diagnostic tests [dFAT, mouse inoculation test (MIT), immunohistochemistry (IHC), RT-PCR and RT-qPCR] and to compare those results obtained under routine laboratory conditions. The results of the present investigation demonstrate that the molecular techniques are more sensitive and may detect low viral load, even though the non-homogeneous viral distribution caused a false-negative result in dFAT. We also observed a usual alteration in antigens distribution among regions of the central nervous system (CNS). By both dFAT and IHC assays, the most reliable CNS structures were thalamus and midbrain. Although this investigation demonstrated diagnostic sensitivity and specificity close to 100 % in all laboratory techniques employed, a dFAT auxiliary test is required for bovine specimens, such as molecular techniques, when there are poor sampling conditions (low viral load combined with unavailability of brainstem structures).


Subject(s)
Cattle Diseases/diagnosis , Clinical Laboratory Techniques/methods , Immunologic Tests/methods , Rabies/diagnosis , Rabies/veterinary , Animals , Brazil , Cattle , Cattle Diseases/virology , Disease Models, Animal , Fluorescent Antibody Technique, Direct/methods , Immunohistochemistry/methods , Mice , Rabies/immunology , Rabies/virology , Rabies virus/immunology , Real-Time Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , Sensitivity and Specificity , Viral Load
3.
Arq. bras. med. vet. zootec. (Online) ; 71(2): 667-676, mar.-abr. 2019. tab, graf
Article in Portuguese | VETINDEX, LILACS | ID: biblio-1011271

ABSTRACT

Objetivou-se avaliar os fatores da composição da dieta que podem interferir no consumo, na digestibilidade, na produção e na composição do leite de vacas mestiças Holandês x Zebu confinadas em condições experimentais no Brasil. A metanálise foi utilizada para o levantamento de trabalhos científicos de domínio público, compreendendo o período de janeiro de 2000 a dezembro de 2015. A base de dados foi constituída por 40 experimentos com 159 tratamentos, envolvendo 735 vacas em lactação. O consumo de nutrientes digestíveis totais (NDT) é comprometido pelo teor de fibra em detergente ácido (FDA) da dieta, relacionando-se com a digestibilidade da matéria seca (MS) e da fibra em detergente neutro (FDN), o que repercute na eficiência alimentar (EA). Observa-se a relação do teor de carboidratos não fibrosos (CNF) da dieta com o teor de lactose e o efeito do teor de MS da dieta e o elevado teor de FDN no consumo em função das características específicas dos tipos de volumosos utilizados. O consumo de NDT da dieta é prejudicado pela digestibilidade da MS e, principalmente, da FDN em função da proporção e das características dos volumosos utilizados na dieta, e esses fatores repercutem na EA, bem como na produção e na composição do leite.(AU)


The aim was to evaluate diet composition factors that may interfere with intake, digestibility, milk yield and composition of crossbred Holstein x Zebus cows confined under Brazilian experimental conditions. The meta-analysis was used for the survey of scientific works of public domain, covering the period from January 2000 to December 2015. The database consisted of 40 experiments with 159 treatments, involving 735 lactating cows. The voluntary intake of total digestible nutrients (TDN) is compromised by the acid detergent fiber (FAD) content of the diet, related to dry matter digestibility (DM) and neutral detergent fiber (NDF), affecting dietary efficiency (DE). The relation of the non-fibrous carbohydrate (NFC) content of the diet with the lactose content and the effect of the DM diet content and the high NDF content on the voluntary intake are specific characteristics of the types of roughage used. Dietary NDT intake is affected by the digestibility of DM and, mainly, NDF, depending on the proportion and characteristics of the nutrients used in the diet, and these factors have an impact on dietary efficiency, as well as on milk composition.(AU)


Subject(s)
Animals , Female , Pregnancy , Cattle , Milk/chemistry , Diet/veterinary , Animal Feed/analysis , Nutrients
4.
Theriogenology ; 119: 1-9, 2018 Oct 01.
Article in English | MEDLINE | ID: mdl-29958134

ABSTRACT

The aim of this work was to evaluate the proteomic changes that occurred in the frozen-thawed bovine spermatozoa after the addition of l-arginine (L-arg) during in vitro sperm capacitation. Aspects related to the sperm capacitation pattern like membrane integrity, mitochondrial activity, sperm motility and vigor, and the sperm proteome were determined. These were respectively assessed by chlortetracycline staining, H342/PI, JC-1, light microscopy, and the proteomic abundance by nUPLC-MS/MS analysis. Frozen-thawed sperm from three Nellore bulls were capacitated in vitro for 3 h in sp-TALP medium supplemented with 20 µg/mL heparin (Control) or with 20 µg/mL heparin plus 1 mM L-arg (L-arg group). Data were subjected to analysis of variance and means compared by SNK test at 5% probability. When compared to Control, the percentage of sperm motility was higher in the L-arg group (P < 0.05). For test data after 3 h of incubation, sperm capacitated with L-arg showed higher membrane integrity and mitochondrial potential when compared to Control (P < 0.05). Moreover, we observed an increase in the percentage of capacitated sperm pattern (P < 0.05). Protein abundance analysis identified 367 proteins. Forty proteins were differentially abundant between Control and L-arg group (P < 0.05), of which 11 were up-regulated, and 29 were down-regulated in L-arg group. In addition, we observed that one protein was uniquely abundant in the L-arg group. Our findings indicate that the addition of L-arg to the culture medium presented a differential protein abundance pattern and increased the bovine frozen-thawed sperm quality and the percentage of capacitated sperm. The proteomic changes observed may be linked to the molecular mechanisms involved in the action of L-arg on the in vitro sperm capacitation of cattle.


Subject(s)
Arginine/pharmacology , Cattle , Gene Expression Regulation/drug effects , Proteome/drug effects , Spermatozoa/drug effects , Animals , Cryopreservation/veterinary , Freezing , Male , Semen Analysis/veterinary , Semen Preservation/veterinary , Sperm Capacitation , Spermatozoa/metabolism
5.
Rev Sci Instrum ; 88(2): 024903, 2017 Feb.
Article in English | MEDLINE | ID: mdl-28249522

ABSTRACT

Specimens subject to periodic heating must be probed for a calibrated temperature response if standard measurements of thermal diffusivity are to be extended to determine thermal conductivity. A variation on two-color pyrometry is developed to measure both the offset and harmonic amplitudes of temperature fluctuations caused by periodic heating. The requisite pyrometric formulae are derived for low amplitude heating using an expansion of the nonlinear thermal emission. Well-defined uncertainties in the temperature values are determined from experimental uncertainties in radiometric measurements. The accuracy demonstrated in this work is better than 2% for the temperature offset and 3%-8% for the fluctuating temperature amplitude.

6.
Arq. bras. med. vet. zootec ; 66(3): 745-756, 06/2014. tab, graf
Article in Portuguese | LILACS | ID: lil-718093

ABSTRACT

O objetivo do presente trabalho foi acompanhar o desenvolvimento imunológico dos neonatos caninos, a fim de avaliar a imunidade celular pela análise dos leucócitos e linfócitos totais e das subpopulações de linfócitos T (CD4+ e CD8+) pela técnica de citometria de fluxo. Foram utilizados 30 cães neonatos de ambos os sexos, sem raça definida, aos três, 10, 17, 24, 31, 38 e 45 dias de idade. A contagem de leucócitos totais aos 45 dias (11.639±3.574) foi significativamente maior que no terceiro dia de idade (8.740±1.812) (P<0,05); não houve diferença entre a contagem total de linfócitos aos 45 dias em relação ao terceiro dia de idade. Quanto às subpopulações de LT CD4+ e LT CD8+, os percentuais de LT CD4+, aos três dias de idade (24,9±16,8 por cento), foram inferiores quando comparados à média entre o 10°, o 24° e o 31°dia (35,5 por cento), e os de CD8+, ao terceiro dia, menores em relação às médias do 10° e do 31° dia de idade. Pode-se concluir que as subpopulações de LT CD4+ e CD8+ sofrem oscilações durante o desenvolvimento pós-natal, sendo estas crescentes em relação aos níveis obtidos aos três dias de idade. A relação CD4+:CD8+ mostrou superioridade para o primeiro tipo celular, sendo que a maior relação entre CD4+ e CD8+ ocorreu no terceiro dia de idade. Com base nos resultados obtidos neste estudo, notaram-se as diferenças semanais nas populações linfocitárias, o que demonstra a dinâmica dessas células durante o período neonatal...


The aim of this study was to evaluate the immune development of newborn canines, evaluating the cellular immunity by analysis of leukocyte and lymphocyte totals and sub-population of T lymphocytes (CD4+ and CD8+) by the flow cytometry technique. Thirty neonatal mongrel dogs of both sexes were used, at 3, 10, 17, 24, 31, 38 and 45 days of age. The total leukocyte count to 45 days (11639±3574/µL) was significantly greater than on the third day of age (8740±1812) (p<0.05); There were no differences between the total count of lymphocytes at 45 days in relation to the third day of age. As for the subpopulations of CD4+ and CD8+, CD4+ cell percentages, at three days of age (24.9±16.8) they were inferior when compared to the average between the 10th, 24th and 31st day (35.5), and CD8+ cells, on the third day, were lower compared with averages from the 10th and 31st day of age. It can be concluded that the subpopulations of CD4+ and CD8+ cells undergo oscillations during development, with growing postnatal levels obtained at three days old. The CD4:CD8 ratio showed superiority to the first cell type, and the largest relationship between CD4+ and CD8+ occurred on the third day of age. Based on the results obtained in this study, the differences noted in lymphocyte populations weekly showed the dynamics of these cells during the neonatal period...


Subject(s)
Animals , Infant, Newborn , Dogs , Animals, Newborn/immunology , Flow Cytometry/veterinary , Leukocyte Count/veterinary , Dogs/growth & development , Immunization, Passive/veterinary
7.
Minerva Stomatol ; 63(3): 77-83, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24632799

ABSTRACT

AIM: The objective of this study was to compare integration of AlloDerm® acellular dermal grafts in animals subjected to non-ablative laser irradiation and animals not exposed to this therapy. METHODS: Standardized AlloDerm® fragments measuring 5 mm² were grafted into the subcutaneous tissue overlying the calvaria in 32 Wistar rats. Laser therapy (685 ηm), at a dose of 4 J/cm2 per session, was applied immediately after surgical intervention and every 48 hours thereafter for a total of four applications. RESULTS: Analysis of histology slides revealed significantly greater edema in the control group. There was no neutrophil infiltration in the laser-irradiated group at any point during the study period, whereas such infiltration was present in control animals at three of the four points of observation. In the laser therapy group, lymphocyte infiltration was observed from day 1, whereas in the control group, it was only apparent from day 3. Vascularization was substantially greater in the control group. In the experimental group, the AlloDerm® graft was completely replaced by fibrous tissue. CONCLUSION: These findings suggest that add-on non-ablative laser therapy is an effective stimulator of healing and graft integration after placement of AlloDerm® acellular dermal grafts.


Subject(s)
Acellular Dermis , Collagen , Low-Level Light Therapy , Skin Transplantation/methods , Wound Healing/radiation effects , Animals , Chemotaxis, Leukocyte/radiation effects , Edema/etiology , Edema/prevention & control , Fibrosis , Graft Survival/radiation effects , Implants, Experimental , Male , Postoperative Complications/etiology , Postoperative Complications/prevention & control , Random Allocation , Rats , Rats, Wistar , Skull , Subcutaneous Tissue/blood supply , Subcutaneous Tissue/pathology
8.
Neuroscience ; 267: 177-86, 2014 May 16.
Article in English | MEDLINE | ID: mdl-24626159

ABSTRACT

In a previous work we found that nitric oxide (NO) and cyclicGMP (cGMP) inhibit glutamatergic synaptic transmission in trigeminal motoneurons (MnV). Here we study the actions of the NO/cGMP signaling pathway on glycinergic synaptic transmission in trigeminal and hypoglossal motoneurons (MnXII) in brain stem slices of neonatal rats. Glycinergic inhibitory postsynaptic currents (IPSCs) were recorded in MnV by stimulation of the supratrigeminal nucleus (SuV) and in MnXII by stimulation of the nucleus of Roller. The NO donor DETA/NONOate (DETA/NO) reduced the amplitude of the IPSC to 58.1±4.2% of control values in MnV. In the presence of YC-1, a modulator of guanylate cyclase that acts as a NO sensitizer, lower and otherwise ineffective concentrations of DETA/NO induced a reduction of the IPSC to 47.2±15.6%. NO effects were mimicked by 8 bromo cyclicGMP (8BrcGMP). They were accompanied by an increase in the paired pulse facilitation (PPF) and in the failure rate of evoked IPSCs. 8BrcGMP did not modify the glycinergic currents elicited by exogenous glycine. In MnXII the IPSCs were also reduced by NO donors and 8BrcGMP to 52.9±6.3% and 45.9±4% of control values, respectively. In these neurons, but not in MnV, we also observed excitatory postsynaptic actions of NO donors. We propose that the differences between the two motor pools may be due to a differential development of the nitrergic system in the two nuclei. Our data show that NO, through its second messenger cGMP, reduces inhibitory glycinergic synaptic transmission in both MnV and MnXII. For MnV, evidence in favor of presynaptic inhibition of glycine release is presented. Given our previous data together with the current results, we propose that the NO/cGMP signaling pathway participates pre- and postsynaptically in the combined regulation of MnV and MnXII activities in motor acts in which they participate.


Subject(s)
Cyclic GMP/metabolism , Glycine/metabolism , Hypoglossal Nerve/cytology , Motor Neurons/physiology , Nitric Oxide/metabolism , Signal Transduction/physiology , Synaptic Transmission/physiology , Trigeminal Nuclei/cytology , Action Potentials/drug effects , Action Potentials/physiology , Animals , Animals, Newborn , Choline O-Acetyltransferase/metabolism , Dose-Response Relationship, Drug , Enzyme Inhibitors/pharmacology , Excitatory Amino Acid Antagonists/pharmacology , Glycine Agents/pharmacology , In Vitro Techniques , NADPH Dehydrogenase/metabolism , Rats , Synaptic Transmission/drug effects
9.
Arq. bras. med. vet. zootec ; 66(1): 219-228, fev. 2014. ilus, tab
Article in Portuguese | LILACS | ID: lil-704027

ABSTRACT

Dois experimentos avaliaram a resposta em ganho de peso de novilhas e a produção de leite de vacas à remoção de folhas apicais e laterais da cana-de-açúcar. Em ambos, os animais foram alimentados individualmente em confinamento total. No Experimento 1, 32 novilhas Girolando (295±45kg de peso vivo) foram blocadas em pares e alocadas a um tratamento por 42 dias, após um período de padronização de 14 dias. Os tratamentos foram: cana integral ou despalhada. O peso vivo e o consumo foram mensurados ao longo do tempo. Os teores de ingredientes e nutrientes na matéria seca (MS) das dietas foram 77,1% de cana, 19,2% de farelo de soja, 1% de ureia, 14,1% de proteína bruta (PB) e 38,8% de fibra em detergente neutro (FDN) na cana despalhada, e 78,6%, 17,9%, 0,9%, 13,7% e 45,3% na cana integral, respectivamente. O ganho diário de peso foi 1,395kg na cana despalhada e 1,125kg na integral (P=0,05). Não houve efeito de tratamento sobre o consumo de MS (P=0,78). A despalha aumentou a taxa de ingestão de alimento (P=0,04) e o consumo diário de matéria orgânica não FDN digestível (P=0,03). No Experimento 2, 14 vacas Holandesas (256±124 dias em lactação) foram blocadas em pares e alocadas a uma sequência dos dois tratamentos em delineamento de reversão simples, com períodos de 21 dias e mensuração da resposta na terceira semana. A dieta com cana despalhada teve 18,4% de cana na MS, 37,6% de silagem de milho, 44% de concentrado baseado em milho, polpa cítrica e farelo de soja, 17,3% de PB e 32,9% de FDN, enquanto a dieta com cana integral teve 18,2%, 37,7%, 44,1%, 16,6% e 34,7%, respectivamente. A despalha tendeu a aumentar a digestibilidade da MS (P=0,06) e o consumo diário de matéria orgânica digestível (P=0,10), sem afetar a produção de leite (18,4kg, P=0,65)....


Two experiments evaluated the responses in weight gain of heifers and milk yield of cows to the removal of the apical and lateral leaves of sugarcane. In both the animals were individually fed in total confinement. In Experiment 1, 32 Holstein x Gir heifers (295±45kg of body weight) were paired, blocked and allocated to a treatment for 42 days, following a 14-day standardization period. Treatments were whole or deleafed sugarcane. Body weight and intake were measured over time. The content of ingredients and nutrients in diets dry matter (DM) were 77.1% sugarcane, 19.2% soybean meal, 1% urea, 14.1% crude protein (CP), and 38.8% neutral detergent fiber (NDF) for deleafed sugarcane, and 78.6%, 17.9%, 0.9%, 13.7%, and 45.3% for whole sugarcane, respectively. Daily weight gain was 1.395kg for deleafed and 1.125kg for whole sugarcane (P=0.05). There was no treatment effect on DM intake (P=0.78). Leaf removal increased the rate of feed intake (P=0.04) and the daily intake of digestible non-NDF organic matter (P=0.03). In Experiment 2, 14 Holsteins (256±124 days of lactation) were paired blocked and allocated to a sequence of two treatments in a cross - over design, with 21 - day periods, and measurement of the response on the third week. The deleafed sugarcane diet had 18.4% sugarcane in DM, 37.6% corn silage, 44% corn, citrus pulp, soybean meal based concentrate, 17.3% CP, and 32.9% NDF, while the whole sugarcane diet had 18.2%, 37.7%, 44.1%, 16.6%, and 34.7%, respectivel y. Deleafing tended to increase DM digestibility (P=0.06) and the daily intake of digestible organic matter (P=0.10), without affecting milk yield (18.4kg, P=0.65). Deleafing s ugarcane increased the weight gain of heifers consuming a high proportion of sug arcane in the diet, but this practice did not induce a positive performance response in late lactating cows consuming 18% sugarcane in the diet.


Subject(s)
Animals , Cattle/classification , Digestion/physiology , Weight Gain/physiology , Mastication , Saccharum
10.
Med Oral Patol Oral Cir Bucal ; 19(2): e184-91, 2014 Mar 01.
Article in English | MEDLINE | ID: mdl-24316700

ABSTRACT

OBJECTIVE: Several studies have focused on the relationship among serotype distribution, ethnical status and geographic populations, and periodontal conditions. Studies that have investigated the prevalence and the distribution of A. actinomycetemcomitans serotypes and the relation between the different serotypes of the bacterium and periodontal status were reviewed. MATERIAL AND METHODS: A systematic literature search for publications regarding the distribution of A. actinomycetemcomitans serotypes in subgingival samples of periodontitis patients and periodontally healthy subjects by employing polymerase chain reaction (PCR) was conducted. RESULTS: From the 85 studies identified in the first analysis, only 12 met all inclusion and exclusion criteria. Clinical isolates from diverse geographic populations with different periodontal conditions were evaluated. Serotypes a, b and c were largely found, and serotype c was the most prevalent. They were isolated from various periodontal conditions, including aggressive periodontitis. CONCLUSIONS: The available literature suggests that serotypes a, b, and c are globally dominant, serotypes d and e are rare, and the prevalence of the most recently identified serotype f is still unknown. It is widely accepted that distribution patterns of A. actinomycetemcomitans vary among subjects of different ethnicity and geographic regions. The correlation of different serotypes with various periodontal conditions remains unclear.


Subject(s)
Aggregatibacter actinomycetemcomitans/classification , Pasteurellaceae Infections , Periodontal Index , Periodontitis/microbiology , Geography , Humans , Serotyping
11.
Neuroscience ; 177: 138-47, 2011 Mar 17.
Article in English | MEDLINE | ID: mdl-21185916

ABSTRACT

In the present work we study the contribution of the chloride channel of the Cystic Fibrosis Transmembrane Regulator (CFTR) in the postsynaptic inhibition of somatic motoneurons during rapid-eye-movement (REM) sleep atonia. Postsynaptic inhibition of motoneurons is partially responsible for the atonia that occurs during REM sleep. Disfacilitation is an additional mechanism that lowers motoneuron excitability in this state. Postsynaptic inhibition is mediated by the release of glycine from synaptic terminals on motoneurons, and by GABA that plays a complementary role to that of glycine. In this work we look in brain stem motoneurons of neonatal rats at a mechanism unrelated to the actions of glycine, GABA or to disfacilitation which depends on the chloride channel of the CFTR. We studied the presence of CFTR by immunocytochemistry. In electrophysiological experiments utilizing whole cell recordings in in vitro slices we examined the consequences of blocking this chloride channel. The effects on motoneurons of the application of glycine, of the application of glibenclamide (a CFTR blocker) and again of glycine during the effects of glibenclamide were studied. Glycine produced an hyperpolarization, a decrease in motoneuron excitability and a decrease in input resistance, all characteristic changes of the postsynaptic inhibition produced by this neurotransmitter. Glibenclamide produced an increase in input resistance and in motoneurons' repetitive discharge as well as a shift in the equilibrium potential for chloride ions as indicated by the displacement of the reversal potential for glycinergic actions. In motoneurons treated with glibenclamide, glycine produced postsynaptic inhibition but this effect was smaller when compared to that elicited by glycine in control conditions. The fact that blocking of the CFTR-chloride channel in brain stem motoneurons influences glycinergic inhibition suggests that this channel may play a complementary role in the glycinergic inhibition that occurs during REM sleep.


Subject(s)
Cystic Fibrosis Transmembrane Conductance Regulator/physiology , Glycine/physiology , Motor Neurons/physiology , Neural Inhibition/physiology , Pons/physiology , Trigeminal Nuclei/physiology , Animals , Animals, Newborn , Motor Neurons/cytology , Organ Culture Techniques , Pons/cytology , Rats , Rats, Wistar , Sleep, REM/physiology , Synaptic Transmission/physiology , Trigeminal Nuclei/cytology
12.
Br J Pharmacol ; 161(4): 899-910, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20860667

ABSTRACT

BACKGROUND AND PURPOSE: The serine and cysteine peptidase inhibitor, BbCI, isolated from Bauhinia bauhinioides seeds, is similar to the classical plant Kunitz inhibitor, STI, but lacks disulphide bridges and methionine residues. BbCI blocks activity of the serine peptidases, elastase (K(iapp) 5.3 nM) and cathepsin G (K(iapp) 160.0 nM), and the cysteine peptidase cathepsin L (K(iapp) 0.2 nM). These three peptidases play important roles in the inflammatory process. EXPERIMENTAL APPROACH: We measured the effects of BbCI on paw oedema and on leucocyte accumulation in pleurisy, both induced by carrageenan. Leucocyte-endothelial cell interactions in scrotal microvasculature in Wistar rats were investigated using intravital microscopy. Cytokine levels in pleural exudate and serum were measured by elisa. KEY RESULTS: Pretreatment of the animals with BbCI (2.5 mg·kg(-1)), 30 min before carrageenan-induced inflammation, effectively reduced paw oedema and bradykinin release, neutrophil migration into the pleural cavity. The number of rolling, adhered and migrated leucocytes at the spermatic fascia microcirculation following carrageenan injection into the scrotum were reduced by BbCI pretreatment. Furthermore, levels of the rat chemokine cytokine-induced neutrophil chemo-attractant-1 were significantly reduced in both pleural exudates and serum from animals pretreated with BbCI. Levels of interleukin-1ß or tumour necrosis factor-α, however, did not change. CONCLUSIONS AND IMPLICATIONS: Taken together, our data suggest that the anti-inflammatory properties of BbCI may be useful in investigations of other pathological processes in which human neutrophil elastase, cathepsin G and cathepsin L play important roles.


Subject(s)
Anti-Inflammatory Agents/pharmacology , Inflammation/drug therapy , Plant Proteins/pharmacology , Animals , Anti-Inflammatory Agents/isolation & purification , Bauhinia/chemistry , Carrageenan , Cathepsin G/antagonists & inhibitors , Cathepsin G/metabolism , Cathepsin L/antagonists & inhibitors , Cathepsin L/metabolism , Cell Adhesion/drug effects , Cell Movement/drug effects , Cytokines/metabolism , Disease Models, Animal , Edema/drug therapy , Edema/physiopathology , Enzyme-Linked Immunosorbent Assay , Humans , Inflammation/physiopathology , Leukocyte Elastase/antagonists & inhibitors , Leukocyte Elastase/metabolism , Leukocytes/drug effects , Leukocytes/metabolism , Male , Microscopy/methods , Plant Proteins/isolation & purification , Rats , Rats, Wistar , Seeds
13.
Br. j. pharmacol ; 161(4): 899-910, Jun 17, 2010.
Article in English | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP, SESSP-IBACERVO | ID: biblio-1061583

ABSTRACT

The serine and cysteine peptidase inhibitor, BbCI, isolated from Bauhinia bauhinioides seeds, is similar to the classical plant Kunitz inhibitor, STI, but lacks disulphide bridges and methionine residues. BbCI blocks activity of the serine peptidases, elastase (Kiapp 5.3 nM) and cathepsin G (Kiapp 160.0 nM), and the cysteine peptidase cathepsin L (Kiapp 0.2 nM). These three peptidases play important roles in the inflammatory process. We measured the effects of BbCI on paw oedema and on leucocyte accumulation in pleurisy, both induced by carrageenan. Leucocyte–endothelial cell interactions in scrotal microvasculature in Wistar rats were investigated using intravital microscopy. Cytokine levels in pleural exudate and serum were measured by elisa.Pretreatment of the animals with BbCI (2.5 mg·kg−1), 30 min before carrageenan-induced inflammation, effectively reduced paw oedema and bradykinin release, neutrophil migration into the pleural cavity. The number of rolling, adhered and migrated leucocytes at the spermatic fascia microcirculation following carrageenan injection into the scrotum were reduced by BbCI pretreatment. Furthermore, levels of the rat chemokine cytokine-induced neutrophil chemo-attractant-1 were significantly reduced in both pleural exudates and serum from animals pretreated with BbCI. Levels of interleukin-1â or tumour necrosis factor-á, however, did not change.Taken together, our data suggest that the anti-inflammatory properties of BbCI may be useful in investigations of other pathological processes in which human neutrophil elastase, cathepsin G and cathepsin L play important roles.


Subject(s)
Animals , Rats , Bauhinia/microbiology , Bradykinin , Cytokines , Plants/immunology , Plant Preparations/antagonists & inhibitors , Pancreatic Elastase , Pleurisy
14.
Cytokine ; 50(3): 335-40, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20231101

ABSTRACT

Previously, we have shown that electroacupuncture (EA) in rats decreased eosinophil infiltration into the pulmonary tissue (PT) and in the bronchoalveolar lavage (BAL) in an experimental model of asthma. Th2 cytokines, leukotriene B4 (LTB4) and nitric oxide (NO) are involved in the asthma inflammatory process. The aim of this study was to verify the effects of EA on these asthma mediators. Male Wistar rats were divided into control (C), immobilized (I), sham acupuncture (SA), and acupuncture (A) groups. All rats were sensitized, and EA treatment using clinical acupuncture points was started 24h after antigen priming. EA was done every other day for 2weeks. Subsequently, animals were challenged by inhalation and sacrificed 24h later. At this time, the BAL and lungs were collected and used to analyze cytokine production, LTB4 and NO. The EA increased IL-1 and IFN-gamma and decreased IL-4, IL-10, NO and LTB4 in the BAL and PT compared to the C and SA groups. The presence of eosinophils in the BAL negatively correlated with IL-1 and IFN-gamma production and positively correlated with IL-4 and IL-10 production. Our results show that the beneficial anti-inflammatory action of EA on asthma is related to the balance of the Thl/Th2 response and the reduction of LTB4 and NO. These results suggest that EA therapy could be an important complementary treatment for asthma.


Subject(s)
Asthma/immunology , Cytokines/immunology , Electroacupuncture , Inflammation/immunology , Leukotriene B4/immunology , Nitric Oxide/immunology , T-Lymphocytes, Helper-Inducer/immunology , Animals , Bronchoalveolar Lavage Fluid/immunology , Eosinophils/immunology , Humans , Male , Rats , Rats, Wistar , Th1 Cells/immunology , Th2 Cells/immunology
15.
Minerva Stomatol ; 58(11-12): 585-91, 2009.
Article in English | MEDLINE | ID: mdl-20027129

ABSTRACT

AIM: The aim of the present study was to compare cephalometric norms of white Brazilian adults with the values described by Tweed, who measured white North American adults. Sexual dimorphism was also evaluated, and male and female facial features were compared. METHODS: Samples were obtained from 40 lateral cephalograms and divided into groups of 20, according to patient's gender. The radiographs were analyzed using the computer-assisted Tweed cephalometric analysis and the 2.0 Radiocef software. Results were analyzed statistically using the Student's t test for genders and ethnicities. The level of significance was set at P< or =0.05. RESULTS: The comparison with Tweed norms revealed that Brazilians have a convex profile and more marked labial tipping of mandibular incisors than North Americans. Sexual dimorphism was found only in linear measurements, which suggests that facial morphology is similar for Brazilian males and females. CONCLUSIONS: Results revealed significant statistical differences in linear and angular cephalometric dimensions between different ethnic groups, which supports the idea that ethnicity should be included in cephalometric analyses.


Subject(s)
Cephalometry , Face/anatomy & histology , Skull/anatomy & histology , White People , Adult , Brazil , Dental Occlusion , Ethnicity , Female , Humans , Male , Radiography , Reference Values , Sex Characteristics , Skull/diagnostic imaging , Young Adult
16.
Transplant Proc ; 37(6): 2791-2, 2005.
Article in English | MEDLINE | ID: mdl-16182811

ABSTRACT

INTRODUCTION: Cirrhosis predisposes to the development of hepatocellular carcinoma (HCC), demanding that these patients undergo follow-up with imaging methods for the early detection of neoplastic nodules. Morphologic study of the explanted livers allows identification of lesions not detected pre-operatively. OBJECTIVE: To assess the frequency and to characterize the HCC found incidentally at pathological evaluation of explanted cirrhotic livers. MATERIAL AND METHODS: Thirty patients had HCC diagnosed in explanted cirrhotic livers. The livers were sectioned at intervals of 0.5 cm. Each detached nodule was selected for microscopic study according to size, color and/or consistency. Retrospective analysis of the clinical records was performed comparing available data from various imaging methods. RESULTS: In 11 patients, 16 HCC had previously been identified in the explants by one or more imaging methods. In the other 19 explanted livers (63.3%), HCC was incidentally found. All HCC identified in 9 patients and some of them in 10 patients were incidentally found, varying from microscopic focuses to 2 cm diameter lesions. They varied from only one (5 cases) to multiple nodules (7 cases). DISCUSSION: Imaging methods may underestimate the number of HCC in cirrhotic livers. Histological study is an essential tool that can early detect tumor nodules not previously detected by imaging methods. In our study, although small, multicentric HCC were identified, illustrating the multifocal nature of the hepatic carcinogenesis. Future studies must correlate these findings with patient outcomes. CONCLUSION: Characterization of explanted cirrhotic livers for HCC is important for previously known and incidentally found lesions.


Subject(s)
Carcinoma, Hepatocellular/pathology , Liver Cirrhosis/epidemiology , Liver Neoplasms/pathology , Liver/pathology , Biopsy , Carcinoma, Hepatocellular/etiology , Hepatitis C/pathology , Humans , Liver Cirrhosis/etiology , Liver Cirrhosis/pathology , Liver Cirrhosis, Alcoholic/pathology , Liver Neoplasms/etiology
17.
Transplant Proc ; 36(4): 958-60, 2004 May.
Article in English | MEDLINE | ID: mdl-15194333

ABSTRACT

OBJECTIVE: To evaluate the impact of infections caused by multiple-drug-resistant (MDR) bacteria on the clinical outcome of liver transplant recipients. METHODS: Retrospective study including all episodes of bacterial infection diagnosed in patients undergoing liver transplantation from January 19, 1999, to June 30, 2002. The diagnosis of bacterial infection required microbiological documentation. Mortality associated with episodes of infection by MDR bacteria was compared to that observed after antibiotic-susceptible bacterial infections. RESULTS: Among 99 patients undergoing liver transplantation during the study period, there were 57 episodes of bacterial infections. Gram-negative bacilli were the predominant etiologic agents (76%) and Pseudomonas aeruginosa was the most frequent bacterial species found in these cases (23 isolates, 28%). Thirty-six episodes of infection (63%) were caused by MDR bacteria. Mean time after transplantation to the diagnosis of infection was 17 days. Mortality associated with episodes of MDR bacterial infections (nine deaths, 25%) was not significantly different from that observed during episodes of antibiotic-susceptible bacteria (five deaths, 24%; P =.92). CONCLUSION: These data suggest that resistance to multiple antimicrobial agents does not have an impact on the mortality associated to bacterial infections in liver transplant recipients.


Subject(s)
Anti-Bacterial Agents , Bacterial Infections/drug therapy , Drug Resistance, Bacterial , Drug Resistance, Multiple , Drug Therapy, Combination/therapeutic use , Liver Transplantation/physiology , Postoperative Complications/microbiology , Bacteria/classification , Bacteria/drug effects , Bacterial Infections/epidemiology , Brazil , Humans , Microbial Sensitivity Tests , Retrospective Studies
18.
Braz. j. biol ; 63(4): 599-615, Nov. 2003. ilus, tab
Article in English | LILACS | ID: lil-355879

ABSTRACT

The pollen spectra of honey samples collected in five apiaries situated near cerrado areas in Minas Gerais State, Brazil, were studied from April 1996 to April 1998 in order to establish the contribution of different plant species in pollen or nectar production The honey samples were characterized by the following native species: Astronium sp., Alternanthera sp., Schinus sp., and Serjanea sp. The pollinic participation percentage of those species was related to the degree of preservation in the areas.


Subject(s)
Animals , Honey , Plants , Pollen , Bees , Brazil , Seasons
19.
Article in English | MEDLINE | ID: mdl-12890550

ABSTRACT

We studied the chronic effect of exercise during water immersion, associated with thermal stress (water temperature at 22, 35 and 40 degrees C) at an intensity of 80% of maximal work load supported in pregnant rats (P) and non-pregnant female rats (NP). P and NP were subdivided into three subgroups according to water temperature during exercise (P22 and NP22; P35 and NP35; P40 and NP40). The animals were submitted to daily swimming sessions of 10-15 min, for 19 days of pregnancy (P) or experimental conditions (NP). Plasma concentration of triglycerides, cholesterol, glucose, total protein, albumin and corticosterone were determined 24 h after the last exercise session. Weight gain and rectal temperature pre- and post-swimming session were also determined. The offspring were examined just after caesarian section on the 20th day of pregnancy to check weight, length and litter size. Pregnant rats showed an increase of triglycerides, reduction of glycemia, total protein and albumin and cholesterol (at 35 degrees C) when compared to non-pregnant animals. Such effects probably lead to an adequate delivery of substrate to the fetus and prepare the mother for lactation. Daily thermal stress did not modify metabolic responses to exercise in pregnant rats. Results also show a deleterious effect on offspring when the mother is exposed daily to extreme temperatures during swimming. These results suggest that water temperature (cold and hot) in swimming have to be considered to avoid damage in fetal development.


Subject(s)
Pregnancy, Animal/physiology , Swimming , Temperature , Water , Animals , Blood Glucose/analysis , Blood Proteins/analysis , Cholesterol/blood , Corticosterone/blood , Female , Litter Size , Pregnancy , Rats , Rats, Wistar , Rectum/physiology , Triglycerides/blood , Weight Gain
20.
Braz J Biol ; 63(4): 599-615, 2003 Nov.
Article in English | MEDLINE | ID: mdl-15029371

ABSTRACT

The pollen spectra of honey samples collected in five apiaries situated near cerrado areas in Minas Gerais State, Brazil, were studied from April 1996 to April 1998 in order to establish the contribution of different plant species in pollen or nectar production The honey samples were characterized by the following native species: Astronium sp., Alternanthera sp., Schinus sp., and Serjanea sp. The pollinic participation percentage of those species was related to the degree of preservation in the areas.


Subject(s)
Honey/analysis , Plants/classification , Animals , Bees , Brazil , Pollen/classification , Seasons
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