ABSTRACT
It was previously demonstrated that the methanol fraction of Sideroxylon obtusifolium (MFSOL) promoted anti-inflammatory and healing activity in excisional wounds. Thus, the present work investigated the healing effects of MFSOL on human keratinocyte cells (HaCaT) and experimental burn model injuries. HaCaT cells were used to study MFSOL's effect on cell migration and proliferation rates. Female Swiss mice were subjected to a second-degree superficial burn protocol and divided into four treatment groups: Vehicle, 1.0% silver sulfadiazine, and 0.5 or 1.0% MFSOL Cream (CrMFSOL). Samples were collected to quantify the inflammatory mediators, and histological analyses were performed after 3, 7, and 14 days. The results showed that MFSOL (50 µg/mL) stimulated HaCaT cells by increasing proliferation and migration rates. Moreover, 0.5% CrMFSOL attenuated myeloperoxidase (MPO) activity and also stimulated the release of interleukin (IL)-1ß and IL-10 after 3 days of treatment. CrMFSOL (0.5%) also enhanced wound contraction, promoted improvement of tissue remodeling, and increased collagen production after 7 days and VEGF release after 14 days. Therefore, MFSOL stimulated human keratinocyte (HaCaT) cells and improved wound healing via modulation of inflammatory mediators of burn injuries.
Subject(s)
Burns , Sapotaceae , Burns/drug therapy , Female , Humans , Keratinocytes , Methanol , Plant Leaves , Proline , Wound HealingABSTRACT
It was previously demonstrated that the methanol fraction of Sideroxylon obtusifolium (MFSOL) promoted anti-inflammatory and healing activity in excisional wounds. Thus, the present work investigated the healing effects of MFSOL on human keratinocyte cells (HaCaT) and experimental burn model injuries. HaCaT cells were used to study MFSOL's effect on cell migration and proliferation rates. Female Swiss mice were subjected to a second-degree superficial burn protocol and divided into four treatment groups: Vehicle, 1.0% silver sulfadiazine, and 0.5 or 1.0% MFSOL Cream (CrMFSOL). Samples were collected to quantify the inflammatory mediators, and histological analyses were performed after 3, 7, and 14 days. The results showed that MFSOL (50 μg/mL) stimulated HaCaT cells by increasing proliferation and migration rates. Moreover, 0.5% CrMFSOL attenuated myeloperoxidase (MPO) activity and also stimulated the release of interleukin (IL)-1β and IL-10 after 3 days of treatment. CrMFSOL (0.5%) also enhanced wound contraction, promoted improvement of tissue remodeling, and increased collagen production after 7 days and VEGF release after 14 days. Therefore, MFSOL stimulated human keratinocyte (HaCaT) cells and improved wound healing via modulation of inflammatory mediators of burn injuries.
Subject(s)
Humans , Female , Burns/drug therapy , Sapotaceae , Proline , Keratinocytes , Plant Leaves , MethanolABSTRACT
ABSTRACT The permanent investigation of new antimycobacterial drugs is necessary for the eradication programs of tuberculosis and other mycobacterium-related diseases. The aim of the present study is to search for new sources of antimycobacterial drugs using plant materials. In this study, 11 plant materials (extracts, essential oils and some fractions) obtained from 4 species of medicinal plants traditionally used as general therapeutics for different illnesses and specifically as treatment of tuberculosis, were evaluated using the microplate resazurin assay against 2 species of the Mycobacterium tuberculosis Complex and 3 nontuberculous mycobacteria. The results showed the hexane extract and the essential oil from fruits of Pterodonemarginatus (Vogel) as potential sources of antimycobacterial drugs against 4 species of tested mycobacteria. The hexane fraction of methanol extract from leaves of Centella asiatica also presented significant mycobacterial growth inhibition, but against M. chelonae only. In conclusion, it was possible to contribute to the antimycobacterial investigations by presenting three new samples of plants with significant antimicrobial activity against four Mycobacteriumspp and suggest future studies about the antimycobacterial properties of fruits from P. emarginatus.
RESUMO A investigação permanente de novas drogas antimicobacterianas é necessária no programa de erradicação da tuberculose e de outras doenças relacionadas com micobactérias. O objetivo deste estudo foi buscar novas fontes de drogas antimicobacterianas usando material vegetal. Neste estudo, 11 materiais de base vegetal (extratos, óleos essenciais e algumas frações) foram avaliados contra 5 espécies de micobactérias. Estes materiais foram obtidos a partir de 4 espécies de plantas medicinais tradicionalmente utilizadas como terapêutica geral para diferentes doenças e, especificamente, no tratamento de tuberculose (Baccharis dracunculifolia, Centella asiatica, Lantana camara, Pterodon emarginatus). Os ensaios foram realizados em microplacas com resazurina contra duas espécies do Complexo Mycobacteriumtuberculosis e 3 espécies de micobactérias não tuberculosas. Os resultados mostraram o extrato hexânico e o óleo essencial de frutos de P.emarginatus como potenciais fontes para drogas antimicobacterianas contra quatro espécies de micobactérias testadas. A fração hexânica do extrato metanólico das folhas de C. asiatica também apresentou significativa inibição do crescimento de micobactérias apenas contra M.chelonae. Em conclusão, foi possível contribuir para as investigações de antimicobacterianos por apresentar três novas amostras de plantas com atividade antimicrobiana significativa contra quatro Mycobacterium spp e sugerir a realização de estudos futuros sobre as propriedades antimicobacterianas de frutos de P. emarginatus.
Subject(s)
/classification , Baccharis/classification , Lantana/classification , Anti-Infective Agents/pharmacology , Plants , Nontuberculous MycobacteriaABSTRACT
BACKGROUND: We recently described the Mycobacterium tuberculosis RD(Rio) genotype, a clonally derived sublineage within the Latin American-Mediterranean (LAM) family. Genetic diversity of M. tuberculosis likely affects the clinical aspects of tuberculosis (TB). Prospective studies that address this issue are scarce and remain controversial. OBJECTIVE: To determine the association of differential clinical features of pulmonary TB with the RD(Rio) M. tuberculosis etiology. METHODS: Culture-proven pulmonary TB patients (n = 272) were clinically evaluated, including history, physical examination, chest X-ray and anti-human immunodeficiency virus serology. Isolates were classified as RD(Rio) or non-RD(Rio) M. tuberculosis by multiplex polymerase chain reaction and further spoligotyped. Clinical and M. tuberculosis genotype data were analyzed. RESULTS: RD(Rio) M. tuberculosis caused disease in 26.5% (72/270) of all TB cases. The LAM genotype, of which RD(Rio) strains are members, was responsible for 46.0% of the TB cases. Demographic data, major signs and symptoms, radiographic presentation, microbiological features and clinical outcomes were not significantly different among patients with TB caused by RD(Rio) and non-RD(Rio) strains. CONCLUSIONS: Disease caused by M. tuberculosis RD(Rio) strains was not clinically distinctive or more severe than disease caused by non-RD(Rio) strains in this series of TB patients. Larger prospective studies specifically designed to disclose differential clinical characteristics of TB caused by specific M. tuberculosis lineages are needed.
Subject(s)
DNA, Bacterial/analysis , Mycobacterium tuberculosis/genetics , Tuberculosis, Pulmonary/microbiology , Adolescent , Adult , Aged , Aged, 80 and over , Brazil/epidemiology , DNA Fingerprinting , Female , Genotype , Humans , Male , Middle Aged , Mycobacterium tuberculosis/isolation & purification , Polymerase Chain Reaction , Prospective Studies , Tuberculosis, Pulmonary/diagnosis , Tuberculosis, Pulmonary/epidemiology , Young AdultABSTRACT
Estimou-se a prevalência de tuberculose em bovinos, e em seus respectivos rebanhos, abatidos em 2009 no estado de Mato Grosso, utilizando como diagnóstico confirmatório o exame bacteriológico e o molecular a partir de fragmentos de tecidos lesionados. Nos sete abatedouros selecionados, detentores de serviço de inspeção federal (SIF), foram inspecionados 41.193 bovinos, sadios ao exame ante mortem, procedentes de 492 rebanhos originários de 85 (60%) municípios mato-grossenses. Um total de 198 carcaças apresentaram lesões suspeitas. Apenas três carcaças (3/198) apresentaram lesões confirmadas como tuberculosas pelos diagnósticos laboratoriais. A prevalência aparente de tuberculose bovina em animais e rebanhos abatidos no estado de Mato Grosso foi de 0,007% [IC 95% = -0,001%; 0,016%] e 0,61% [IC 95% = -0,08%; 1,30%], respectivamente. O estado do Mato Grosso possui, naturalmente, um status sanitário considerado de baixa prevalência.(AU)
The prevalence of bovine tuberculosis in cattle, and its herds, slaughtered in 2009 in the state of Mato Grosso, Brazil, was estimated using bacteriological analysis and molecular test, from fragments of injured tissues as well as direct DNA templates. 41,193 cattle, which appeared healthy in the ante mortem examination, from seven selected slaughterhouses, under Brazilian federal inspection services (SIF), were inspected. The animals were from 492 herds located in 85 (60%) different cities of Mato Grosso. A total of the 198 carcasses had suspicious lesions. Three carcasses (3/198) had lesions that were found to be tuberculous in laboratory diagnosis. The apparent prevalence of bovine tuberculosis in the total number of animals and in herds slaughtered in Mato Grosso was 0.007% [IC 95% = -0.001%; 0.016%] and 0.61% [IC 95% = -0.08%; 1.30%], respectively. The sanitation status demonstrated in Mato Grosso indicates the progress in this state toward the eradication of bovine tuberculosis.(AU)
Subject(s)
Animals , Cattle , Cattle/microbiology , Mycobacterium bovis , Clinical Laboratory Techniques/veterinary , Lymph Nodes/pathology , Inflammation/veterinary , Polymerase Chain Reaction/veterinary , Tissue Array Analysis/veterinaryABSTRACT
Estimou-se a prevalência de tuberculose em bovinos, e em seus respectivos rebanhos, abatidos em 2009 no estado de Mato Grosso, utilizando como diagnóstico confirmatório o exame bacteriológico e o molecular a partir de fragmentos de tecidos lesionados. Nos sete abatedouros selecionados, detentores de serviço de inspeção federal (SIF), foram inspecionados 41.193 bovinos, sadios ao exame ante mortem, procedentes de 492 rebanhos originários de 85 (60%) municípios mato-grossenses. Um total de 198 carcaças apresentaram lesões suspeitas. Apenas três carcaças (3/198) apresentaram lesões confirmadas como tuberculosas pelos diagnósticos laboratoriais. A prevalência aparente de tuberculose bovina em animais e rebanhos abatidos no estado de Mato Grosso foi de 0,007% [IC 95% = -0,001%; 0,016%] e 0,61% [IC 95% = -0,08%; 1,30%], respectivamente. O estado do Mato Grosso possui, naturalmente, um status sanitário considerado de baixa prevalência.
The prevalence of bovine tuberculosis in cattle, and its herds, slaughtered in 2009 in the state of Mato Grosso, Brazil, was estimated using bacteriological analysis and molecular test, from fragments of injured tissues as well as direct DNA templates. 41,193 cattle, which appeared healthy in the ante mortem examination, from seven selected slaughterhouses, under Brazilian federal inspection services (SIF), were inspected. The animals were from 492 herds located in 85 (60%) different cities of Mato Grosso. A total of the 198 carcasses had suspicious lesions. Three carcasses (3/198) had lesions that were found to be tuberculous in laboratory diagnosis. The apparent prevalence of bovine tuberculosis in the total number of animals and in herds slaughtered in Mato Grosso was 0.007% [IC 95% = -0.001%; 0.016%] and 0.61% [IC 95% = -0.08%; 1.30%], respectively. The sanitation status demonstrated in Mato Grosso indicates the progress in this state toward the eradication of bovine tuberculosis.
ABSTRACT
AIMS: To compare three decontamination methods applied to paucibacillary samples for primary isolation of Mycobacterium bovis from suspect lesions. Tuberculosis caused by Myco. bovis is an important infectious disease of cattle in Brazil and also has zoonotic potential. Although a national campaign based on testing and slaughtering cattle has achieved good results, there is a strong need to develop better diagnostic methods to identify cattle with recent infections harbouring few bacilli. METHODS AND RESULTS: A dairy herd (274 adult crossbred cows) located in the state of Rio de Janeiro was tested for tuberculosis with both single intradermal tuberculin test and comparative intradermal tuberculin test. Reactive cows (n=27, 9.8%) were slaughtered and suspect lesions were collected (one sample per cow). Samples considered paucibacillary (based on microscopy) were decontaminated with 0.75% hexadecylpyridinium chloride (HPC), 4% sodium hydroxide (Petroff) or 6% sulphuric acid. Using these methods, 10, five and six, respectively, of the 27 samples yielded positive cultures. Overall, Myco.bovis was isolated from 14 of 24 cows. Although the HPC method resulted in isolation of more Myco.bovis strains than either Petroff or sulphuric acid methods (P=0.015), it did not result in the recovery of Myco.bovis from all samples. However, using both HPC and 6% sulphuric acid methods for decontamination was possible to identify 13 of 14 (92·9%) of infected cows. CONCLUSIONS: At least two methods should be used concurrently for primary isolation of Myco. bovis from bovine tissues, particularly for paucibacillary samples. SIGNIFICANCE AND IMPACT OF THE STUDY: Detection of low numbers of Myco.bovis in tissue is an important goal in optimizing the detection of bovine tuberculosis and should assist in identification of infected cattle, in particular, those with few Myco.bovis bacilli. This was apparently the first study comparing three decontamination methods for the detection of Myco.bovis in paucibacillary samples from naturally infected cattle.
Subject(s)
Decontamination/methods , Mycobacterium bovis/isolation & purification , Tuberculosis, Bovine/diagnosis , Animals , Cattle , Cetylpyridinium/chemistry , Female , Lung/microbiology , Lung/pathology , Mycobacterium bovis/genetics , Tuberculin Test , Tuberculosis, Bovine/microbiologyABSTRACT
An interferometer that measures the refractive index changes due to bacterial metabolism is described. The apparatus permits simultaneous and real time measurement of bacterial growth in several samples of slowly growing mycobacteria. The error sources are discussed and the sensitivity of the apparatus is tested. For the species Mycobacterium bovis BCG and M. smegmatis, a relation between refractive index change and bacterial concentration is determined experimentally and the time constant of bacterial growth is measured.
Subject(s)
Energy Metabolism/physiology , Interferometry/instrumentation , Mycobacterium tuberculosis/isolation & purification , Mycobacterium tuberculosis/physiology , Refractometry/instrumentation , Calibration , Cell Proliferation , Equipment Design , Equipment Failure Analysis , Interferometry/standards , Refractometry/methods , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
Phenotypic characteristics, antimicrobial susceptibility, and genetic relationships were analyzed in 107 Staphylococcus aureus isolates recovered from cows with subclinical mastitis in Southeastern Brazil. Thirteen different biochemical patterns were detected among isolates. A predominant pattern represented by about 54% of the isolates was distributed among several herds. Isolates of distinct phenotypic profiles were also detected within a herd. Susceptibility to ampicillin, cefotaxime, cephalotin, chloramphenicol, erythromycin, gentamycin, kanamycin, nitrofurantoin, norfloxacin, ofloxacin, oxacillin, penicillin, rifampin, sulfamethoxazole-trimethoprim, tetracycline, trimethoprim, and vancomycin, determined by the disk diffusion method, was observed in 44.9% of isolates. On the other hand, 55.1, 7.4, and 2.8% of the strains were resistant to ampicillin/penicillin, tetracycline, and erythromycin, respectively. Genetic diversity was analyzed by pulsed-field gel electrophoresis using SmaI as the restriction enzyme. All isolates could be typed by pulsed-field gel electrophoresis, which identified 16 types and 24 subtypes. Type A and its subtypes comprised 54.2% of all isolates and were recovered from 6 of the 9 herds analyzed. Other types and subtypes were also found in multiple herds. Although multiple types and subtypes were found within a specific herd, a predominant type was frequently observed.
Subject(s)
Drug Resistance, Bacterial , Mastitis, Bovine/microbiology , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/metabolism , Animals , Brazil , Cattle , Electrophoresis, Gel, Pulsed-Field , Female , Fermentation , Genetic Variation , Genotype , Mastitis, Bovine/drug therapy , Microbial Sensitivity Tests , Milk/microbiology , Phenotype , Staphylococcus aureus/geneticsABSTRACT
Este trabalho teve como objetivo avaliar o ciclo reprodutivo do bagre de água doce Parauchenipterus striatulus, na represa de Ribeiräo das Lajes, Rio de Janeiro. Foram descritos aspectos morfológicos dos estádios de maturaçäo gonadal bem como variaçöes no índice gonadossomático, relaçäo peso: comprimento, fator de condiçäo gonadal. A amostragem dos peixes foi feita bimestralmente entre abril de 1996 e maio de 1997 com o uso de redes de espera de 100m de comprimento, 4m de altura e com malha entre 25 e 65mm de distância entrenós. Os 339 indivíduos capturados ao longo do período de amostragem corresponderam a 40 por cento do total de peixes. Cinco estádios de maturaçäo gonadal foram determinados por meio de análise macroscópica para ambos os sexos: imaturo, maturaçäo I, maturaçäo II, maduro, desovado (fêmeas) e esvaziado (machos). O índice gonadossomático foi mais elevado entre outubro e março para ambos os sexos, sugerindo amplo período de desova enquanto o fator de condiçäo atingiu seu máximo entre fevereiro a junho, mostrando uma relaçäo inversa entre essas duas características. A relaçäo peso-comprimento para machos foi de W=0,0095L3,0862 sobrescrito e fêmeas de W=0,0116L3,0126 sobrescrito, näo tendo sido encontradas diferenças significativas entre os coeficientes de alometria para ambos os sexos
Subject(s)
Animals , Male , Female , Catfishes , Fishes , ReproductionABSTRACT
Glycosphingolipids were extracted from hyphae of Fusarium solani and from an unnamed Fusarium species, and were purified by silica and Iatrobead column chromatography. Their structures were determined by compositional analysis, nuclear magnetic resonance spectroscopy, gas chromatography/mass spectrometry and by fast atom bombardment mass spectrometry of the native and peracetylated materials, which defined their sugar, long-chain base and fatty acid compositions. The locations of the double bonds in the bases were established by 2D NMR spectroscopy and by novel mass spectrometric approaches, including collisional activation of the protonated and lithium-cationized glycosphingolipids, and of the sphingadienene-derived fragment ion at m/z 276. From these results we propose that the structures of the glycosphingolipids from F. solani and Fusarium sp. are N-2'-hydroxyoctadecanoyl-1-O-beta-D-glucopyranosyl-9-methyl-4, 8-sphingadienine and N-2'-hydroxyoctadecenoyl-1-O-beta-D-glucopyranosyl-9-methyl-4, 8-sphingadienine, respectively.