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Estudaram-se as alterações tonométricas, paquimétricas e de comprimento axial em cães com glaucoma submetidos à ablação uveal intravítrea. Foram avaliados 13 olhos irreversivelmente cegos de cães que apresentavam glaucoma crônico unilateral, nos quais realizou-se a ablação uveal intra-vítrea, por meio de injeção na câmara vítrea de 0,5ml de sulfato de gentamicina (40mg/ml) associado a 0,3ml de fosfato de dexametasona (4mg/ml). As mensurações da pressão intra-ocular (Po), espessura corneana e eixo axial com tonometria de aplanação, paquimetria ultra-sônica e ultra-sonografias modos A e B foram realizadas no dia da ablação (M0) e após uma (M1), quatro (M4), oito (M8), 12 (M12), 24 (M24) e 48 semanas (M48). A Po diminuiu significativamente em todos os momentos em relação ao M0, com aumento da espessura corneana ao longo do experimento. Nas avaliações ultra-sonográficas notou-se diminuição significativa do bulbo ocular a partir de M4, com medidas ultra-sonográficas modo A significativamente maiores que as do modo B. O procedimento foi efetivo na redução da Po e na diminuição do eixo axial, demonstrando ser viável no controle do glaucoma crônico em olhos irreversivelmente cegos, e ser uma alternativa à enucleação e inserção de prótese ocular(AU)
Changes in tonometry, pachymetry and globe axial length in glaucomatous dogs submitted to intravitreal uveal ablation were studied. Thirteen irreversible blind canine eyes were evaluated. They presented unilateral chronic glaucoma, in which intravitreal uveal ablation was performed, through vitreous chamber injection of 0.5ml of gentamicin sulfate solution (40mg/ml) with 0.3ml of dexametazon dissodic phosphate (4mg/ml). The measurements of intraocular pressure (IOP), corneal thickness and globe axial length with aplanation tonometry, ultrasound pachymetry and A and B-mode ultrasound scan were made on the day of the procedure (M0), and on the next day (M1), four (M4), eight (M8), 12 (M12), 24 (M24) and 48 weeks (M48) after. The IOP decreased on all moments in relation to M0, with corneal thickness increasing throughout the experiment. The ultrasonographic evaluations revealed significant globe decrease from M4, with A-scan measurements significantly higher than B-scan. The procedure was effective on intraocular pressure reduction and in shortening the axial length, showing to be feasible to control chronic glaucoma in irreversible blind eyes, as well as an alternative to enucleation and intraocular prothesis(AU)
Assuntos
Animais , Masculino , Feminino , Cães , Tonometria Ocular/métodos , Tonometria Ocular/veterinária , Ultrassonografia/métodos , Glaucoma/fisiopatologia , Gentamicinas/administração & dosagem , Cirurgia Filtrante/métodos , Cães , Ultrassonografia/veterináriaRESUMO
Estudaram-se as alterações tonométricas, paquimétricas e de comprimento axial em cães com glaucoma submetidos à ablação uveal intravítrea. Foram avaliados 13 olhos irreversivelmente cegos de cães que apresentavam glaucoma crônico unilateral, nos quais realizou-se a ablação uveal intra-vítrea, por meio de injeção na câmara vítrea de 0,5ml de sulfato de gentamicina (40mg/ml) associado a 0,3ml de fosfato de dexametasona (4mg/ml). As mensurações da pressão intra-ocular (Po), espessura corneana e eixo axial com tonometria de aplanação, paquimetria ultra-sônica e ultra-sonografias modos A e B foram realizadas no dia da ablação (M0) e após uma (M1), quatro (M4), oito (M8), 12 (M12), 24 (M24) e 48 semanas (M48). A Po diminuiu significativamente em todos os momentos em relação ao M0, com aumento da espessura corneana ao longo do experimento. Nas avaliações ultra-sonográficas notou-se diminuição significativa do bulbo ocular a partir de M4, com medidas ultra-sonográficas modo A significativamente maiores que as do modo B. O procedimento foi efetivo na redução da Po e na diminuição do eixo axial, demonstrando ser viável no controle do glaucoma crônico em olhos irreversivelmente cegos, e ser uma alternativa à enucleação e inserção de prótese ocular
Changes in tonometry, pachymetry and globe axial length in glaucomatous dogs submitted to intravitreal uveal ablation were studied. Thirteen irreversible blind canine eyes were evaluated. They presented unilateral chronic glaucoma, in which intravitreal uveal ablation was performed, through vitreous chamber injection of 0.5ml of gentamicin sulfate solution (40mg/ml) with 0.3ml of dexametazon dissodic phosphate (4mg/ml). The measurements of intraocular pressure (IOP), corneal thickness and globe axial length with aplanation tonometry, ultrasound pachymetry and A and B-mode ultrasound scan were made on the day of the procedure (M0), and on the next day (M1), four (M4), eight (M8), 12 (M12), 24 (M24) and 48 weeks (M48) after. The IOP decreased on all moments in relation to M0, with corneal thickness increasing throughout the experiment. The ultrasonographic evaluations revealed significant globe decrease from M4, with A-scan measurements significantly higher than B-scan. The procedure was effective on intraocular pressure reduction and in shortening the axial length, showing to be feasible to control chronic glaucoma in irreversible blind eyes, as well as an alternative to enucleation and intraocular prothesis
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Animais , Masculino , Feminino , Cães , Cirurgia Filtrante/métodos , Cães , Gentamicinas/administração & dosagem , Glaucoma/fisiopatologia , Tonometria Ocular/métodos , Tonometria Ocular/veterinária , UltrassonografiaRESUMO
The identification of 163 strains of Mycobacterium bovis by polymerase chain reaction (PCR) and microbiological tests was carried out on 252 tuberculous-like lesions (TLLs) collected from slaughtered cattle in south-east Brazil. This study compared the usefulness of three genotyping techniques, IS6110-restriction fragment length polymorphism (RFLP), polymorphic guanine-cytosine-rich sequence (PGRS)-RFLP and direct repeat (DR)-spoligotyping, as applied to M. bovis isolates. Based on IS6110-RFLP genotyping we selected a group of 23 isolates containing more than one IS6110 copy, along with 16 samples containing one IS6110 copy from different geographical areas, evenly distributed among dairy (eight) and beef cattle (eight). These selected isolates were analysed by PGRS-RFLP and DR-spoligotyping genotyping. Dairy cattle (17%) display a higher frequency of multiple IS6110 copies than beef cattle (10%). A comparison between the genotype data obtained fails to show a correlation between the main clusters found by the three techniques. However, the clustering of each genotyping procedure revealed that the majority of strains are closely related. The RFLP-PGRS patterns showed a sizable group (20.5%) containing a 5.5 kb fragment and the predominant spoligotype is similar to that from the BCG vaccine strain. Unexpectedly, four strains (2.4%) showed drug resistance to 0.2 microg/ml isoniazid and 20 microg/ml ethionamide, but none of them was resistant to rifampicin or other antibiotics tested.
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Técnicas de Tipagem Bacteriana/veterinária , Mycobacterium bovis/classificação , Mycobacterium bovis/genética , Polimorfismo de Fragmento de Restrição , Tuberculose Bovina/microbiologia , Animais , Brasil , Bovinos , Análise por Conglomerados , Impressões Digitais de DNA/veterinária , Farmacorresistência Bacteriana/genética , FilogeniaRESUMO
The pyrazinamidase gene coding for the enzyme that activates the bactericidal drug pyrazinamide contains a polymorphic site that is preserved in Mycobacterium bovis. We synthesized two sets of primers, one encompassing a 180 bp fragment, and the second spanning a 726 bp fragment including the full pncA gene. Following PCR of Mycobacterium tuberculosis and M. bovis samples, it is possible to discriminate by this polymorphism between these species by digestion with Eco065 I. Digestion of the 180 bp fragment results in two fragments of 101 and 79 bp, specific for M. tuberculosis. Alternatively, digestion of the 726 bp fragment yields three fragments of 452, 165 and 109 bp for M. tuberculosis, but only two fragments of 561 and 165 bp for M. bovis.
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Amidoidrolases/genética , Mycobacterium bovis/genética , Mycobacterium tuberculosis/genética , Polimorfismo de Fragmento de Restrição , Sequência de Bases , Humanos , Mycobacterium bovis/isolamento & purificação , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose Pulmonar/diagnóstico , Tuberculose Pulmonar/microbiologiaRESUMO
Large-scale vaccination with BCG, the live attenuated strain of Mycobacterium bovis, is being adopted around the world, although sporadic complications have occurred after the procedure. Lymphadenopathy is not uncommon especially in babies under one year (0.73 percent of vaccinated infants), but the swelling subsides within 2 months in most cases, with no medical or surgical treatment. Brazil adopted BCG vaccination program earlier in the seventies and by 1995 more than 96 percent of the infant population received this immunization. We report here the occurrence of lymphadenopathy in a two-year-old child vaccinated with the Brazilian BCG strain. The diagnosis was made using a lymph node biopsy and intestinal aspirates that yielded a positive mycobacterial culture. The isolate was resistant to isoniazid, rifampicin, pyrazinamide and thiophen-2-carbonic acid hydrazide, sensitive to streptomycin, ethambutol, and p-nitrobenzoic acid, and reacted positively to cyclo-serine and negatively to niacin. The pncA gene involved in bacterial activation of pyrazinamide contains in M. bovis a point mutation that renders pyrazinamidase unable to catalyze drug activation. Therefore, this polymorphism is a good option for developing methods to differentiate M. bovis and M. tuberculosis. Taking advantage of this difference we further analyzed the isolates by single-stranded conformation polymorphism electrophoresis of DNA following PCR of the pncA gene. The isolate identity was confirmed by RFLP electrophoretic analysis of the amplified fragment following Eco065I digestion, which selectively cleaves M. tuberculosis DNA. From this result it is proposed that RFLP of pncA gene represents an alternative for differential diagnosis of M. bovis.
Assuntos
Humanos , Masculino , Pré-Escolar , Vacina BCG , Linfadenite , Mycobacterium bovis , DNA Bacteriano , Linfonodos , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de RestriçãoRESUMO
Large-scale vaccination with BCG, the live attenuated strain of Mycobacterium bovis, is being adopted around the world, although sporadic complications have occurred after the procedure. Lymphadenopathy is not uncommon especially in babies under one year (0.73% of vaccinated infants), but the swelling subsides within 2 months in most cases, with no medical or surgical treatment. Brazil adopted BCG vaccination program earlier in the seventies and by 1995 more than 96% of the infant population received this immunization. We report here the occurrence of lymphadenopathy in a two-year-old child vaccinated with the Brazilian BCG strain. The diagnosis was made using a lymph node biopsy and intestinal aspirates that yielded a positive mycobacterial culture. The isolate was resistant to isoniazid, rifampicin, pyrazinamide and thiophen-2-carbonic acid hydrazide, sensitive to streptomycin, ethambutol, and p-nitrobenzoic acid, and reacted positively to cyclo-serine and negatively to niacin. The pncA gene involved in bacterial activation of pyrazinamide contains in M. bovis a point mutation that renders pyrazinamidase unable to catalyze drug activation. Therefore, this polymorphism is a good option for developing methods to differentiate M. bovis and M. tuberculosis. Taking advantage of this difference we further analyzed the isolates by single-stranded conformation polymorphism electrophoresis of DNA following PCR of the pncA gene. The isolate identity was confirmed by RFLP electrophoretic analysis of the amplified fragment following Eco065I digestion, which selectively cleaves M. tuberculosis DNA. From this result it is proposed that RFLP of pncA gene represents an alternative for differential diagnosis of M. bovis.
Assuntos
Vacina BCG/efeitos adversos , Linfadenite/microbiologia , Mycobacterium bovis/isolamento & purificação , Amidoidrolases/genética , Pré-Escolar , DNA Bacteriano/análise , Humanos , Linfonodos/microbiologia , Linfadenite/diagnóstico , Masculino , Mycobacterium bovis/genética , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de RestriçãoRESUMO
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Three hundred and twenty 30-week old White Leghorn cockerels were housed in individual cages and distributed in a completely randomized factorial design of 5 x 3, with five oil sources (sunflower, soybean, canola, linseed and fish/soybean) and three levels of antioxidant (30, 200 and 400 mg of vitamin E/kg). The aim of this study was to evaluate the effect of the ratio of omega6: omega3 fatty acids by the inclusion of different oil sources and of dietary supplementation with vitamin E on the reproductive performance of cockerels. The use of the fish/soybean combination determined the lowest total antioxidant status of the semen. However, the addition of vitamin E to the fish/soybean-oil-based diet resulted in a linear increase in semen volume, motility and sperm vigour in the 38th week and again in the 52nd week for motility and for sperm vigour and fertility rate in the periods from 50-53 and 41-53 weeks of age. The use of canola oil in the diet resulted in the highest fertility rate during 50-53 and 41-53 weeks of life. Animals receiving the soybean oil based diet showed the smallest fertility rate in the range from 50-53 weeks of age and concomitantly the highest level of cholesterol in the spermatozoa in the range from 47-51 weeks. An interaction between the vitamin E level and soybean oil was verified by the linear increase in motility and sperm vigour at 38 weeks of age. Later, the contrary was shown by a linear reduction in fertility in the periods from 44-46, 47-49 and 41-53 weeks of age. Cockerels that had been fed on the sunflower-oil-based diet showed the highest content of saturated fatty acids in the spermatozoa from 48-51 weeks. An interaction effect was observed between the vitamin E level and sunflower oil shown by a linear increase in the content of saturated fatty acids in the spermatozoa and a linear reduction in the C18: 1omega9, C18 :2 omega6 and PUFA (C18 : 2omega6 + C20 : 4omega6) contents in the spermatozoa at 48-51 weeks and in sperm volume at 52 weeks of age.
Assuntos
Antioxidantes/administração & dosagem , Galinhas/fisiologia , Ácidos Graxos Ômega-3/administração & dosagem , Ácidos Graxos Ômega-6/administração & dosagem , Fertilidade/efeitos dos fármacos , Vitamina E/administração & dosagem , Animais , Suplementos Nutricionais , Relação Dose-Resposta a Droga , Óleos de Peixe/administração & dosagem , Masculino , Distribuição Aleatória , Sêmen/efeitos dos fármacos , Sêmen/fisiologia , Óleo de Soja/administração & dosagem , Contagem de Espermatozoides/veterinária , Motilidade dos Espermatozoides , Espermatogênese/efeitos dos fármacos , Espermatozoides/química , Espermatozoides/fisiologiaRESUMO
Diagnosis of the Mycobacterium tuberculosis complex by direct PCR of mediastinal lymphnode DNA and microbiological tests were compared in cattle suspicious of bearing tuberculous-like lesions detected during slaughter. The PCR procedure applied on DNA samples (n=54) obtained by adding alpha -casein into the thiocyanate extraction mix was positive in 70% of the samples. PCR confirmed the identification of 23 samples (100%) that grew in culture, 9 samples (60%) that failed to grow in culture, plus 6 (37.5%) samples that resulted in growth of bacterial contaminants. Genotyping by IS6110-RFLP and DR-spoligotyping analysis of seven samples revealed the presence of several polimorphisms. Seven of the isolates contained multiple copies of IS6110, thus defining the existence of five singular genotypes.
Assuntos
Linfonodos/microbiologia , Mycobacterium bovis/genética , Animais , Bovinos , DNA Bacteriano/análise , Genótipo , Mycobacterium bovis/isolamento & purificação , Reação em Cadeia da Polimerase , Polimorfismo Genético , Polimorfismo de Fragmento de Restrição , Tuberculose Bovina/microbiologiaRESUMO
Diagnosis of the Mycobacterium tuberculosis complex by direct PCR of mediastinal lymphnode DNA and microbiological tests were compared in cattle suspicious of bearing tuberculous-like lesions detected during slaughter. The PCR procedure applied on DNA samples (n=54) obtained by adding alpha -casein into the thiocyanate extraction mix was positive in 70 percent of the samples. PCR confirmed the identification of 23 samples (100 percent) that grew in culture, 9 samples (60 percent) that failed to grow in culture, plus 6 (37.5 percent) samples that resulted in growth of bacterial contaminants. Genotyping by IS6110-RFLP and DR-spoligotyping analysis of seven samples revealed the presence of several polimorphisms. Seven of the isolates contained multiple copies of IS6110, thus defining the existence of five singular genotypes
Assuntos
Animais , Bovinos , Linfonodos/microbiologia , Mycobacterium bovis/genética , DNA Bacteriano , Genótipo , Mycobacterium bovis/isolamento & purificação , Reação em Cadeia da Polimerase , Polimorfismo Genético , Polimorfismo de Fragmento de Restrição , Tuberculose Bovina/patologiaRESUMO
We report a case of a carotid artery bifurcation aneurysm which ruptured into a silvian fissure arachnoid cyst. In the review of the literature, only three cases were before reported. We discuss about uncommon clinical findings, the surgical aspects and the associations among the lesions.
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Aneurisma Roto/complicações , Cistos Aracnóideos/complicações , Aneurisma Intracraniano/complicações , Hemorragia Subaracnóidea/etiologia , Adulto , Aneurisma Roto/diagnóstico por imagem , Aneurisma Roto/cirurgia , Cistos Aracnóideos/diagnóstico por imagem , Cistos Aracnóideos/cirurgia , Angiografia Cerebral , Humanos , Aneurisma Intracraniano/diagnóstico por imagem , Aneurisma Intracraniano/cirurgia , Masculino , Tomografia Computadorizada por Raios XRESUMO
Natural killer (NK) cells play an important role in immune surveillance against tumors. The present work aimed to study the cytotoxic activity of NK cells and T cell subsets in peripheral blood of 13 patients with primary tumors in central nervous system (CNS). As controls 29 healthy subjects with the age range equivalent to the patients were studied. The methods employed were: a) determination of cytotoxic activity of NK cells towards K562 target cells, evaluated by single cell-assay; b) enumeration of CD3+ lymphocytes and their CD4+ and CD8+ subsets defined by monoclonal antibodies; c) the identification of tumors were done by histologic and immunochemistry studies. The results indicated that adults and children with tumor in CNS display reduced percentage of total T cells, helper/inducer subset and low helper/suppressor ratio. The cytotoxic activity of NK cells was decreased in patients with CNS tumors due mainly to a decrease in the proportion of target-binding lymphocytes. These results suggest that cytotoxic activity of NK cells may be affected by the immunoregulatory disturbances observed in patients with primary tumors in CNS.
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Neoplasias do Sistema Nervoso Central/imunologia , Células Matadoras Naturais/imunologia , Subpopulações de Linfócitos T/imunologia , Adolescente , Adulto , Neoplasias do Sistema Nervoso Central/sangue , Criança , Pré-Escolar , Citotoxicidade Imunológica , Feminino , Humanos , Imunidade Celular , Masculino , Pessoa de Meia-Idade , Subpopulações de Linfócitos T/químicaRESUMO
A simple method was developed for DNA extraction of Mycobacterium bovis in milk and further detection of the bacterium by polymerase chain reaction (PCR). Milk previously seeded with M. bovis was used as the starting material. The procedure involved overnight digestion of a milk sample with proteinase K at 56 degrees C and phenol extraction, followed by ethanol precipitation and PCR. The amplification pattern obtained was analyzed with primers BW8-BW9 which amplify a 248 bp in strains of M. bovis. By using the BW8-BW9 primers, 10(3) CFU were detected on silver-stained PAGE gels. The procedure was validated by PCR analysis of milk in tuberculin-positive animals. It is anticipated that this method can be used for routine diagnosis of M. bovis in milk samples.
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DNA Bacteriano/isolamento & purificação , Leite/microbiologia , Mycobacterium bovis/isolamento & purificação , Animais , Bovinos , DNA Bacteriano/genética , Endopeptidase K , Mycobacterium bovis/genética , Reação em Cadeia da Polimerase/métodosRESUMO
We describe the use of a plant cysteine proteinase isolated from latex of Carica candamarcensis as a protective agent during isolation of bacterial DNA following growth in culture of these cells. Between 100 to 720 units of proteinase (1 µg = 6 units) afforded good DNA protection when incubated with various kinds of microorganisms. Agarose gel electrophoresis showed that the resulting DNA was similar in size to DNA preparations obtained by treatment with proteinase K. The viability of the resulting material was checked by PCR amplification using species-specific primers. After standing at room temperature (25oC) for 35 days, the enzyme lost 10 percent of its initial activity. The enzyme stability and good yield of DNA suggest the use of this proteinase as an alternative to proteinase K
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Animais , Cisteína Proteases , DNA/isolamento & purificação , Endopeptidase K , Substâncias Protetoras/farmacologia , DNA Bacteriano/isolamento & purificação , Eletroforese em Gel de Ágar , Endopeptidase K/farmacologia , Leptospira , Reação em Cadeia da PolimeraseRESUMO
There are many reports supporting a self-limitation mechanism involved with hypermetabolic response after severe cranial injury. It was proposed a study with severe head injury patients, in three stages of the evolution. The first 7 days after admission (moment 1-M1), the second three days latter (M2) and the last 7 days after the first (M3). Among male patients with severe head injury, attended between January 1992 and December 1993 in University Hospital of Botucatu, UNESP, were selected 28 male patients, with Glasgow severity scale between 4 and 6, with pO2 < 70 mm Hg, weighting 60 kg or more. Among these patients, 6 finished the study, including analysis of the excretion of N, acute phase proteins, glycemia, triglycerides and amine nitrogen. During the study there were no changes in nitrogen balance and there was a decrease in protein C-reative. Glycemia tends to fall within two weeks after injury. The authors make some considerations about possible mechanisms involved in brain modulation associated with the period of dependence of hypermetabolism and hypercatabolism after closed brain injury. There are some evidences that the brain responds to head trauma with a gobal non specific way, which tends to be reorganized beyond the first two weeks after lesion. The study does not show any influence of the type and severity of head trauma.