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1.
Exp Parasitol ; 243: 108408, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36336025

RESUMO

As per 20th livestock census, India possessing 193.46 million cattle and 109.85 million buffaloes in organized and unorganized sectors. These animals are suffering from tick infestation almost throughout the year causing both direct and indirect losses. Repeated use of chemical acaricides for tick management resulted in the establishment of acaricide resistant tick populations, insecticide residues in livestock products and environmental pollution. In the present study, analysis of the data generated during 1990-2021 revealed pooled prevalence of infestations in cattle and buffaloes was 53% [95% CI = 47-60%] and 38% [95% CI = 28-49%], respectively. Based on peripheral blood smear examination the prevalence of theileriosis, babesiosis and anaplasmosis in cattle is estimated as 17% [95% CI = 12-24%], 4% [95% CI = 3-6%] and 5% [95% CI = 3-7%], respectively, while in buffalo the prevalence of infection is estimated as 7% [95% CI = 2-21%], 2% [95% CI = 0-5%] and 8% [95% CI = 2-36%],respectively.To estimate economic impact, both direct (reduction in milk production, cost of treatment, leather depreciation) and indirect (milk loss and treatment cost) losses were taken into consideration. Loss of milk production was predicted as 13.91, 56.91 and 85.34L/cross-bred cow/lactation in low, moderate and high tick infestation conditions, respectively. Whereas, 20.10, 7.01L milk/buffalo/lactation in Hyalomma spp. and Rhipicephalus spp. infestation was estimated. Similarly,the estimated loss of milk production due to clinical theileriosis, babesiosis and anaplasmosis was 57.96, 30.96 and 59.22L, respectively. The cumulative (milk loss, treatment cost and leather loss) loss due to tick infestation was calculated as 46199.31 million INR (USD595.07 million) while due to TBDs 14877.15 million INR (USD191.15 million) = 61076.46 million INR (USD787.63 million). The data provided base line information for the policy maker to develop strategies at government level so that the significantly high cumulative loss of 787.63 million USD due to ticks and tick borne diseases (TTBDs) can be minimized.


Assuntos
Acaricidas , Anaplasmose , Babesiose , Doenças dos Bovinos , Rhipicephalus , Theileriose , Infestações por Carrapato , Doenças Transmitidas por Carrapatos , Animais , Feminino , Bovinos , Infestações por Carrapato/epidemiologia , Infestações por Carrapato/veterinária , Theileriose/epidemiologia , Búfalos , Doenças dos Bovinos/epidemiologia , Doenças Transmitidas por Carrapatos/epidemiologia , Doenças Transmitidas por Carrapatos/veterinária , Gado
2.
Exp Parasitol ; 242: 108369, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36058254

RESUMO

Fasciola gigantica faces a series of threats from various free radicals produced by the host immune system during its invasion through the abdominal cavity and establishment in the bile duct of ruminants, limiting the fluke viability. The role of the superoxide radical produced by Muzaffarnagari sheep immune effector cells against F. gigantica newly excysted juveniles (NEJs) is highlighted in this study, as is the critical role of superoxide dismutase enzyme (SOD) in dismutation of superoxide radicals derived from host immune effector cells in vitro. Three concentrations of the ovine immune effector cells viz. 2.5, 5, and 10 × 106 cells were tested for their ability to induced cytotoxic killing of the parasite. All the three cell concentrations caused significant (p < 0.01) cytotoxic killing of NEJs in comparison to the control groups. Also, reduction of the immune effector cell concentration directly correlates with the NEJs killing. Attachment of immune effector cells to the parasite tegument in the presence of anti-F. gigantica antibodies was found to be critical in inducing NEJs killing via antibody-dependent cell-mediated cytotoxicity (ADCC). However, the addition of SOD greatly inhibits cytotoxic killing of NEJs, demonstrating the importance of SOD enzyme in fluke survival and parasite evasion of the host immunity. Thus, F. gigantica SOD warrants a promising candidate for immunoprophylactic studies in ruminants against the tropical liver fluke.


Assuntos
Fasciola hepatica , Fasciola , Fasciolíase , Ovinos , Animais , Superóxidos , Superóxido Dismutase
3.
J Parasit Dis ; 45(2): 330-335, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34295030

RESUMO

Trypanosoma evansi is a flagellated, extracellular haemoprotozoan parasite infecting a wide range of mammalian hosts including dromedaries, cattle, equines and dogs cause disease surra. Carrier animals with sub-clinical infection cause significant monetary losses to livestock holders and therefore detection of infection status using molecular diagnostic techniques becomes important in order to control the disease. In the current study cattle, buffalo, goat, pig and dog samples from three northeastern states of India-Assam, Mizoram and Tripura were screened to determine the prevalence of surra. A total of 1702 samples including 795 from Assam, 678 from Mizoram and 229 from Tripura were screened by CATT/T. evansi test out of which 16.8%, 27.1% and 22.3% samples in respective states were found to have antibodies against T. evansi. DNA detection of T. evansi by PCR amplification targeting VSG gene revealed the molecular prevalence of surra in Assam, Mizoram and Tripura as 8.5%, 7.5% and 4.4% respectively. The analysis of amplified partial VSG sequences showed 99% similarity within an animal species whereas 86-94% similarity was observed among different species of animals revealing the homogeneity. The study established the prevalence of surra in different species of animals in the three northeastern states of India-Assam, Mizoram and Tripura and this study is the first report of T. evansi infection in pig and goat from India. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12639-021-01392-z.

4.
J Parasit Dis ; 45(1): 244-251, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33746410

RESUMO

Surra, a haemoprotozoan parasitic disease even in subclinical form poses a challenge in terms of diagnosis and management to animal health practitioners and policy makers as well; eventually imparting financial loss to the livestock holders. A systematic study was designed to assess the seroprevalence of surra in cattle and associated climatic risk factors, by collecting 480 serum samples across the eight districts of Mizoram during 2017-2019. The apparent and true seroprevalence detected by card agglutination test was 37.08% (CI at 95%: 32.88-41.49) and 36.59% (CI at 95%: 32.4-40.99) whereas by recombinant Variable Surface Glycoprotein based indirect ELISA was 41.88% (CI at 95%: 37.5-46.3) and 40.35% (CI at 95%: 36.02-44.76) respectively. Climate parameters which influence vector population were extracted from their respective database and were correlated with seroprevalence data. Linear discriminant analysis revealed that air temperature, relative humidity and diurnal temperature range, leaf area index and soil moisture as significant risk factors discriminating seropositive and seronegative data sets classified by indirect ELISA. This study is the first report on seroprevalence of surra in cattle of Mizoram and the situation demands deployment of intervention strategies in order to assess the endemicity of the disease and thereby preventing the economic losses.

5.
Vet Parasitol ; 266: 63-66, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30736948

RESUMO

Trypanosoma evansi, a haemo-flagellated protozoan parasite causes chronic wasting disease in a wide range of animals. For its diagnosis, blood smear examination is useful in clinical cases for direct identification of the parasite but in latent infection the carrier animals are difficult to screen out by conventional blood smear test. Harboring low level of parasites and showing no symptom, the carrier animals for surra can act as a source of infection. The level of parasitaemia fluctuates, especially during latent infection; moreover the antibodies which are not found early in the infection may persist even after recovery or chemotherapy. In the present study a double antibody sandwich ELISA exploring, monoclonal antibodies and hyperimmune serum, raised against recombinant variable surface glycoprotein has been developed to detect circulating trypanosome antigens. The developed antigen detection ELISA (Ag-ELISA) was evaluated using 652 blood samples collected from cattle, buffalo, equine and camel. The statistical analysis of the data showed diagnostic sensitivity and specificity at 97.4% and 96.4% respectively, with a positive-negative cut-off OD value >0.28. Furthermore, the detection limit of the assay was found to 7.15 trypanosomes per mL. The present finding revealed that the developed assay can be exploited as a potential diagnostic test in the detection of circulating trypanosome antigens and also can be used as a population screening test for multiple animal species for detection of active infection for further treatment and control of the disease.


Assuntos
Antígenos de Protozoários/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Glicoproteínas de Membrana/imunologia , Proteínas de Protozoários/imunologia , Trypanosoma/imunologia , Tripanossomíase/veterinária , Animais , Anticorpos Monoclonais/imunologia , Antígenos de Protozoários/sangue , Búfalos/parasitologia , Camelus/parasitologia , Bovinos/parasitologia , Cavalos/parasitologia , Glicoproteínas de Membrana/genética , Proteínas Recombinantes/imunologia , Sensibilidade e Especificidade , Tripanossomíase/diagnóstico
6.
J Immunol Methods ; 460: 87-92, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30056943

RESUMO

Trypanosoma evansi, a haemoflagellated protozoan parasite, is responsible for chronic as well as the acute debilitating disease called surra in a wide range of herbivores and carnivores including domestic and wild animals. Since the parasite is having wide host range, there is a need for diagnostic test which can detect the T. evansi specific antibody in different species of animals for generating sero-surveillance data. In the present study we developed and evaluated competitive enzyme immunoassay using monoclonal antibodies (MAbs) raised against recombinant variable surface glycoprotein (rVSG) of T. evansi. The immunoreactivity of the developed MAbs (IgG3-subtype) was evaluated by immunoblot as well as ELISA and subsequently used in the development and standardization of competitive ELISA (C-ELISA). Further, the serological data generated from the C-ELISA using reference samples constituting true positive or surely infected (35), true negative (45), sero-positive (225) and sero-negative (215) samples and was analyzed statistically. The true positivity/negativity was determined by thin blood smear examination and diagnostic PCR assay, While, seropositivity/seronegativity of the reference samples was determined through standard reference tests. The data showed the diagnostic sensitivity of 92.6% and specificity of 96.4% with Cohen's kappa value of 0.88. In order to determine the utility of C-ELISA in detecting T. evansi antibodies in different species of animals, the assay was further evaluated with 1361 field sera sample comprising bovine, horse, donkey and camel. Since the C-ELISA described herein has showed high sensitivity and specificity, this single test can be explored in the sero-surveillance of T. evansi in a wide range of animals.


Assuntos
Anticorpos Monoclonais Murinos/química , Anticorpos Antiprotozoários/química , Antígenos de Protozoários , Proteínas de Protozoários , Trypanosoma , Tripanossomíase , Animais , Antígenos de Protozoários/genética , Antígenos de Protozoários/imunologia , Búfalos , Camelus , Bovinos , Ensaio de Imunoadsorção Enzimática , Equidae , Cavalos , Immunoblotting , Camundongos , Camundongos Endogâmicos BALB C , Proteínas de Protozoários/genética , Proteínas de Protozoários/imunologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Trypanosoma/genética , Trypanosoma/imunologia , Tripanossomíase/diagnóstico , Tripanossomíase/imunologia , Tripanossomíase/veterinária
7.
J Helminthol ; 90(5): 577-82, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26343353

RESUMO

Helminth infections in the mithun Bos frontalis, including the liver fluke Fasciola gigantica, hepato-gastric amphistomes Explanatum explanatum, Paramphistomum epiclitum and Calicophoron calicophorum, and the cestodes Echinococcus granulosus and E. ortleppi were studied in north-east India over a 2-year period from 2012 to 2014. Cystic echinococcosis caused by E. granulosus and E. ortleppi was found to be highly prevalent in the mithun, with E. ortleppi being reported for the first time. Molecular markers, including the internal transcribed spacer 2 (ITS-2), 28S rDNA and mitochondrial NADH dehydrogenase sub-unit1 (nad1) were used to confirm the identification of the trematode and cestode species.


Assuntos
Biodiversidade , Cestoides/isolamento & purificação , Infecções por Cestoides/veterinária , Ruminantes/parasitologia , Trematódeos/isolamento & purificação , Infecções por Trematódeos/veterinária , Animais , Cestoides/classificação , Cestoides/genética , Infecções por Cestoides/epidemiologia , Infecções por Cestoides/parasitologia , DNA de Helmintos/genética , DNA Ribossômico/genética , DNA Espaçador Ribossômico/genética , Marcadores Genéticos , Índia , NADH Desidrogenase/genética , Prevalência , RNA Ribossômico 28S/genética , Trematódeos/classificação , Trematódeos/genética , Infecções por Trematódeos/epidemiologia , Infecções por Trematódeos/parasitologia
8.
Singapore Med J ; 49(1): e12-4, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18204752

RESUMO

A five-year-old boy presented with progressive weight gain with effort intolerance and nocturnal symptoms suggesting obstructive sleep apnoea. A clinical diagnosis of Prader-Willi Syndrome was made. As the initial radiography and computed tomography suggested a foreign body, bronchoscopy was done under general anaesthesia and impacted peanuts were removed from the left main bronchus. His symptoms resolved instantly and the patient was asymptomatic at six months follow-up. This report highlights the need to consider foreign body aspiration as a cause for dyspnoea in children with Prader-Willi Syndrome. The report also focuses on the need to adopt strategies that prevent foreign body aspiration and choking in patients with Prader- Willi Syndrome.


Assuntos
Corpos Estranhos , Síndrome de Prader-Willi/complicações , Apneia Obstrutiva do Sono/complicações , Apneia Obstrutiva do Sono/diagnóstico , Traumatismos Torácicos/diagnóstico , Obstrução das Vias Respiratórias , Brônquios/patologia , Broncografia , Criança , Diagnóstico Diferencial , Humanos , Inalação , Masculino , Radiografia Torácica , Tomografia Computadorizada por Raios X , Traqueia/patologia
9.
Glycoconj J ; 24(1): 49-55, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17115276

RESUMO

Transendothelial migration of monocytes followed by their differentiation into macrophages involves interaction of monocytes with subendothelial matrix. The influence of extracellular matrix on monocyte-macrophage differentiation was studied using an in vitro model system with human PBMC maintained on different matrix protein substrata. Upregulation of macrophage specific marker activities such as endocytosis of modified proteins, changes in expression of cell surface antigen, and production of matrix metalloproteinases was studied. Cells maintained on Fibronectin (Fn) showed significantly higher rate of endocytosis and production of MMP2 and MMP9 when compared to other matrix protein substrata. Immunoblot analysis, ELISA, and zymography showed that Fn-dependent upregulation of MMPs was blocked by antibodies to alpha(5)beta(1) integrin indicating that the Fn effect was mediated by integrins. The Fn effect on mo-mPhi was blocked by genistein and herbimycin. As monocytes differentiate to macrophages there was an increase in the rate of production of Fn. These results indicate the influence of the microenvironment of the cell, particularly Fn, on mo-mPhi differentiation and integrin-mediated downstream signaling through focal adhesion kinase and Src type tyrosine kinase is involved in this.


Assuntos
Diferenciação Celular , Fibronectinas/metabolismo , Macrófagos/citologia , Macrófagos/metabolismo , Monócitos/citologia , Regulação para Cima , Acetilação/efeitos dos fármacos , Anticorpos/farmacologia , Benzoquinonas/farmacologia , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Endocitose/efeitos dos fármacos , Imunofluorescência , Genisteína/farmacologia , Humanos , Lactamas Macrocíclicas/farmacologia , Macrófagos/efeitos dos fármacos , Metaloproteinases da Matriz/genética , Naftalenos/farmacologia , Rifabutina/análogos & derivados , Soroalbumina Bovina/metabolismo , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos
10.
Plant Dis ; 87(3): 247-251, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30812755

RESUMO

Agroinfection of bipartite geminiviruses is routinely done by mixing two Agrobacterium strains that independently harbor partial tandem repeats of DNA A and DNA B. We report here an improved agroinfection method for bipartite geminiviruses that utilizes one strain of Agrobacterium that harbors DNA A and DNA B partial tandem repeats on two compatible replicons. A cointegrate vector, pGV2260∷pGV1.3A, with the partial tandem repeat of Mungbean yellow mosaic virus-Vi (MYMV-Vi) DNA A and a binary vector, pGA1.9B, with the partial tandem repeat of MYMV-Vi DNA B gave an agroinfection efficiency of 24% when harbored in two Agrobacterium strains and an efficiency of 61% when harbored in one Agrobacterium strain. A combination of binary vectors, pGA1.9A with MYMV-Vi DNA A partial tandem repeat and pGA1.9B with DNA B partial tandem repeat, gave an agroinfection efficiency of 74% when harbored in two strains. But pGA1.9A and pPZP1.9B (a partial tandem repeat of DNA B), when present in the same Agrobacterium strain, gave 100% agroinfection. Accumulation of viral DNA was shown by Southern blotting. The single-strain method using two compatible replicons consistently gave 100% agroinfection efficiency.

11.
Am J Ophthalmol ; 132(6): 872-4, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11730651

RESUMO

PURPOSE: To assess the acute effects of sildenafil citrate (VIAGRA) on the intraocular pressure (IOP) of patients with chronic open-angle glaucoma. DESIGN: This was a double-blind, randomized, placebo-controlled, crossover study, in which 15 patients received a single oral dose of sildenafil 100 mg or matching placebo on two separate occasions. METHODS: Fifteen subjects aged 63 +/- 14 years (mean +/- SD) with bilateral chronic open-angle glaucoma were administered a single oral dose of sildenafil 100 mg or matching placebo on two separate occasions at least 3 days apart. IOP was measured in both eyes by Goldmann ap-planation tonometry at baseline and then at 1-5 hours after dosing. Brachial artery systolic and diastolic blood pressures were determined by sphygmomanometry, and heart rate was also monitored at baseline and 1-5 hours after dosing. RESULTS: Compared with placebo, no statistically or clinically significant change in IOP was detected after a single dose of sildenafil 100 mg (P =.20). Moreover, no significant change in mean systemic blood pressure (P =.12) or heart rate (P =.72) was detected after treatment with sildenafil. CONCLUSION: At the maximum therapeutic dose of 100 mg, sildenafil did not produce any significant acute change in IOP in men with chronic open-angle glaucoma. This information is of importance for patients with glaucoma receiving sildenafil for treatment of erectile dysfunction.


Assuntos
3',5'-GMP Cíclico Fosfodiesterases/antagonistas & inibidores , Glaucoma de Ângulo Aberto/complicações , Pressão Intraocular/efeitos dos fármacos , Inibidores de Fosfodiesterase/uso terapêutico , Piperazinas/uso terapêutico , Pressão Sanguínea , Doença Crônica , Estudos Cross-Over , Método Duplo-Cego , Disfunção Erétil/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , Inibidores de Fosfodiesterase/efeitos adversos , Piperazinas/efeitos adversos , Purinas , Citrato de Sildenafila , Sulfonas , Tonometria Ocular
12.
Atherosclerosis ; 159(2): 333-41, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11730813

RESUMO

Blood monocytes (mo) on transendothelial migration interact with extracellular matrix components (ECM) and differentiate into macrophages (m(phi)), which play an important role in both physiological, and pathological conditions, particularly, atherosclerosis. In order to study whether modification of ECM such as non-enzymatic glycation occurring in diabetes influences mo-m(phi) differentiation, an in vitro system using isolated human peripheral blood mononuclear cells (PBMC) maintained on non-enzymatically glycated COL I (NEG COL I) was used. M(phi) specific functions such as receptor mediated endocytosis of modified proteins, production of m(phi) specific 92 and 72 kDa matrix metalloproteinases (MMPs), expression of surface antigen and loss of myeloperoxidase (MPO) activity were assessed. Endocytosis of 125[I] acetyl BSA was significantly higher in cells maintained on NEG COL I than those on COL I. Kinetic analysis revealed that the rate of uptake of modified BSA and production of MMPs by cells maintained on NEG COL I were higher than those on COL I suggesting a faster rate of differentiation of cells maintained on modified substrata. FACS analysis of the expression of surface antigen showed that the rate of down-regulation of monocyte specific CD14 and the rate of up-regulation of m(phi) specific CD71 were high in cells maintained on NEG COL I. These results suggest that the interaction of monocyte with non-enzymatically glycated matrix protein in the vessel wall may result in faster rate of induction of mo-m(phi) differentiation leading to foam cell formation, a critical early event in the initiation and development of atherosclerosis.


Assuntos
Antígenos de Diferenciação/fisiologia , Colágeno/metabolismo , Matriz Extracelular/metabolismo , Macrófagos/fisiologia , Monócitos/fisiologia , Arteriosclerose/fisiopatologia , Diferenciação Celular/fisiologia , Células Cultivadas , Endocitose/fisiologia , Citometria de Fluxo , Humanos , Macrófagos/imunologia , Monócitos/imunologia , Peroxidase/metabolismo , Probabilidade , Sensibilidade e Especificidade
13.
Biochim Biophys Acta ; 1540(1): 50-8, 2001 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-11476894

RESUMO

Human peripheral blood mononuclear cells (PBMC) upon transendothelial migration interact with subendothelial matrix components and differentiate into macrophages. In order to study whether the shape of the cells as dictated by the extracellular matrix can influence monocyte-macrophage (mo-m(phi)) differentiation, human PBMC were maintained in vitro on a three dimensional collagen I (COL I) lattice and studied for various macrophage specific functions, viz. endocytosis of [(125)I]acetyl bovine serum albumin (BSA), expression of specific cell surface antigens and expression of matrix metalloproteinases (MMPs). The cells maintained in three dimensional COL gel exhibited a higher rate of endocytosis of [(125)I]acetyl BSA than those on COL-coated plastic. FACS analysis showed that the mean fluorescence intensity (MFI) corresponding to monocyte specific LPS receptor CD14 was significantly decreased while MFI corresponding to macrophage specific transferrin receptor CD71 was significantly increased in cells maintained in vitro on three dimensional COL gel compared to two dimensional COL substrata. Expression of macrophage specific MMPs (gelatinase A and gelatinase B) was significantly high in cells maintained on COL gel than on COL I-coated plastic. Appearance of 67 kDa gelatinase in the COL gel suggested that induction as well as activation of MMPs occur when cells are maintained in a three dimensional environment. These results indicate that monocytes undergo a rapid rate of differentiation when maintained in vitro on three dimensional COL I lattice suggesting that apart from the chemical nature of the matrix, the shape of the cells as provided by the matrix also influences mo-m(phi) differentiation.


Assuntos
Técnicas de Cultura de Células/métodos , Colágeno , Monócitos/citologia , Antígenos CD/análise , Antígenos de Diferenciação de Linfócitos B/análise , Diferenciação Celular , Endocitose , Matriz Extracelular , Gelatinases/análise , Humanos , Radioisótopos do Iodo , Receptores de Lipopolissacarídeos/análise , Macrófagos/imunologia , Macrófagos/fisiologia , Monócitos/imunologia , Monócitos/fisiologia , Receptores da Transferrina , Soroalbumina Bovina/metabolismo
14.
Am J Ophthalmol ; 131(6): 751-5, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11384572

RESUMO

PURPOSE: Sildenafil citrate induces vasodilation by enhancing the smooth muscle relaxant effects of nitric oxide. We have previously reported that nitrate compounds, a different group of nitric oxide-mediated vasodilators used mainly for the treatment of ischemic cardiac diseases, produce an increase in optic nerve head circulation and retinal venous vasodilation. The purpose of the present investigation was to evaluate the effect of sildenafil on ocular circulation. METHODS: In a double-blind, randomized, crossover trial, 15 healthy male volunteers received 100-mg doses of sildenafil citrate (Viagra; Pfizer, Inc, New York, New York) or matching placebo on 2 separate days. Laser Doppler flowmetry was used to assess foveolar choroidal and optic nerve rim circulatory parameters. Measurements were obtained in one eye at baseline, 1 hour, and 5 hours after dosing. Blood pressure and intraocular pressure were monitored, and perfusion pressure was calculated. RESULTS: Mean optic nerve head blood flow measurements at baseline, 1 hour, and 5 hours were 11.6 +/- 2.2 arbitrary units (+/- SD), 12.5 +/- 2.8, and 12.1 +/- 2.4 after sildenafil and 11.9 +/- 2.5, 12.6 +/- 3.1, and 13.0 +/- 3.0 after placebo, respectively. When compared with placebo, no significant change in mean blood pressure, intraocular pressure, perfusion pressure, or choroidal or optic nerve circulatory parameters were observed after sildenafil treatment. The power to detect a 20% change in optic nerve head and choroidal blood flow after sildenafil was approximately 90%. CONCLUSIONS: In comparison with placebo, no significant change in optic nerve rim or foveolar choroidal blood flow was observed after treatment with sildenafil. This suggests that nitrate compounds and sildenafil may differentially affect ocular circulation. Furthermore, no significant effects on intraocular pressure, systemic blood pressure, or ocular perfusion pressure were detected after sildenafil treatment.


Assuntos
Olho/irrigação sanguínea , Inibidores de Fosfodiesterase/farmacologia , Piperazinas/farmacologia , Vasodilatadores/farmacologia , Adulto , Pressão Sanguínea/efeitos dos fármacos , Corioide/irrigação sanguínea , Estudos Cross-Over , Método Duplo-Cego , Humanos , Pressão Intraocular/efeitos dos fármacos , Masculino , Disco Óptico/irrigação sanguínea , Purinas , Fluxo Sanguíneo Regional/efeitos dos fármacos , Citrato de Sildenafila , Sulfonas
15.
Int J Radiat Oncol Biol Phys ; 50(2): 465-72, 2001 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-11380235

RESUMO

PURPOSE: The purpose of this study was to assess the need for obtaining weekly complete blood count (CBC) values and to identify the pattern of changes in CBC during regional conventional fractionated radiotherapy. METHODS AND MATERIALS: A retrospective analysis of CBC data on 299 adult cancer patients who received definitive conventional radiotherapy to head and neck (n = 95), chest (n = 96), and pelvis (n = 108) was performed. Temporal patterns and magnitude of change in white blood cells, neutrophils, lymphocytes, and platelets during radiotherapy were examined. RESULTS: There were statistically significant declines in all counts, albeit not clinically significant. Notable differences between disease sites were found. The greatest weekly interval change in counts occurred during the first week of radiotherapy for all groups of patients. The mean WBC nadir values during treatment were 5.8 for head & neck, 6.8 for chest, and 5.4 for pelvis. The nadirs for all counts occurred toward the middle-to-end of radiotherapy. Lymphocytes were found to be more sensitive to radiotherapy than other leukocyte subcomponents. CONCLUSION: Our study suggests that weekly CBC monitoring is not necessary for all patients undergoing standard fractionated radiotherapy. Baseline blood counts may be used to determine an optimal schedule for monitoring CBCs in patients receiving conventional radiation alone. Reduced monitoring of CBC may result in significant financial savings.


Assuntos
Plaquetas/efeitos da radiação , Linfócitos/efeitos da radiação , Neoplasias/sangue , Neoplasias/radioterapia , Neutrófilos/efeitos da radiação , Adulto , Plaquetas/citologia , Fracionamento da Dose de Radiação , Humanos , Contagem de Leucócitos , Linfócitos/citologia , Neutrófilos/citologia , Contagem de Plaquetas , Estudos Retrospectivos
16.
Drug Metabol Drug Interact ; 16(3): 159-71, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-11116751

RESUMO

A major metabolic pathway for triamterene (a potassium sparing diuretic) is aromatic hydroxylation followed by sulphate conjugation. Diflunisal (a salicylate anti-inflammatory agent) also undergoes sulphate conjugation of its phenolic group as a major pathway. We investigated the possible effect of diflunisal on the elimination of triamterene (competition for phenolic sulphonation) in six healthy volunteers by studying the disposition of single doses of triamterene (100 mg) taken alone and in the presence of steady-state levels of diflunisal. Diflunisal coadministration (500 mg b.i.d.) had no effect on the pharmacokinetics of triamterene itself. However, plasma AUC of p-hydroxytriamterene sulphate was greater (4.6 times), and its renal clearance lower (0.24 times), in the presence of diflunisal. There was no change in the formation clearance or protein binding of p-hydroxytriamterene sulphate in the presence of diflunisal. The data point to competition for renal excretory pathways rather than sulphonation capacity. This interaction could have clinical relevance since p-hydroxytriamterene sulphate is pharmacologically active.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Inibidores de Ciclo-Oxigenase/farmacologia , Diflunisal/farmacologia , Diuréticos/farmacocinética , Triantereno/farmacocinética , Adulto , Cromatografia Líquida de Alta Pressão/métodos , Diuréticos/sangue , Diuréticos/urina , Interações Medicamentosas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Triantereno/sangue , Triantereno/urina
17.
J Muscle Res Cell Motil ; 15(6): 646-58, 1994 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-7706421

RESUMO

Three recombinant chicken gizzard caldesmon (CaD) baculovirus vectors that contained the full-length CaD codon sequence (Pv1CaD), the full-length CaD codon sequence and a six-histidine tag at the 5'-end (pBlueBacHisCaD), or the full-length CaD codon sequence and an extra six-histidine codon sequence at the 3'-end (PvlHisCaD) were constructed. Spodoptera frugiperda (Sf9) cells transfected with these constructs overexpressed full-length CaD, yielding 2, 20, and 50 micrograms per 10(6) cells for pBlueBacHisCaD, PvlHisCaD, and PvlCaD, respectively. Time course assays for the expressed proteins demonstrated that the optimum harvest time was 36 h postinfection. Immunofluorescence microscopy revealed PvlCaD localized on the plasma membrane of Sf9 cells at 24 h postinfection and distributed throughout the cytoplasm at 36-48 h postinfection. Analysis of the purified recombinant full-length CaD revealed most of the characteristics of the authentic CaD, including (a) an electrophoretic mobility corresponding to 125 kDa, (b) heat stability, (c) binding to actin, tropomyosin-actin, myosin, and calmodulin, (d) ability to inhibit actin-activated ATP hydrolysis by smooth muscle myosin, and (e) ability of Ca(2+)-calmodulin to reverse the inhibition. A CaD mutant with a deletion of 159 amino acids from the carboxyl terminus of the full-length CaD was also expressed at high levels in Sf9 cells. However, this mutant showed a decreased ability to bind to actin, tropomyosin-actin, and calmodulin, whereas the myosin binding was unaffected; actin-activated ATP hydrolysis by smooth muscle myosin was not inhibited by this mutant.


Assuntos
Proteínas de Ligação a Calmodulina/biossíntese , Proteínas Recombinantes de Fusão/biossíntese , Actinas/metabolismo , Animais , Baculoviridae , Sequência de Bases , Calmodulina/metabolismo , Proteínas de Ligação a Calmodulina/química , Proteínas de Ligação a Calmodulina/genética , Proteínas de Ligação a Calmodulina/isolamento & purificação , Galinhas/genética , Clonagem Molecular , Códon/genética , Vetores Genéticos , Proteínas de Membrana/análise , Microscopia de Fluorescência , Dados de Sequência Molecular , Mutagênese , Miosinas/metabolismo , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/isolamento & purificação , Spodoptera , Tropomiosina/metabolismo
18.
J Biol Chem ; 269(22): 15803-7, 1994 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-8195235

RESUMO

A single-headed monomeric myosin (myosin I) was isolated from pig urinary bladder smooth muscles and purified to homogeneity. Myosin I from smooth muscle is composed of a 110-kDa heavy chain and three 17-kDa light chains. The heavy chain from smooth muscle myosin I does not cross-react with the antibody against conventional myosin (myosin II) from smooth muscle, but it does show antigenic similarity to adrenal medulla myosin I heavy chain. The light chain from smooth muscle myosin I is similar to calmodulin in molecular weight, amino acid composition, and migration on SDS-polyacrylamide gel electrophoresis in the presence of Ca2+. The high salt ATPase activity of myosin I in the presence of CaCl2 is higher than that in K(+)-EDTA. Smooth muscle actin causes a 5-10-fold activation of the Mg-ATPase activity of myosin I. In the presence of Ca2+, exogenous calmodulin enhances the actin-activated ATPase activity of myosin I, and the increased activity is associated with the binding of exogenous calmodulin to myosin I heavy chain. A maximum of 4 mol of light chains/mol of myosin I heavy chain is observed in the presence of exogenous calmodulin. Caldesmon, a calmodulin/actin-binding protein, inhibits the actin-activated ATPase activity of myosin I. This inhibition is reversed by exogenous calmodulin in the presence of Ca2+. The actin activation of myosin I ATPase exhibits around 50% Ca2+ sensitivity in the presence of exogenous calmodulin. When caldesmon is bound to actin, Ca2+ sensitivity is increased to 80% in the presence of calmodulin. Therefore, smooth muscle caldesmon, which is thought to play a role in the regulation of actin activation of myosin II, also regulates the actin activation of myosin I ATPase in smooth muscle.


Assuntos
ATPase de Ca(2+) e Mg(2+)/metabolismo , Proteínas de Ligação a Calmodulina/metabolismo , Calmodulina/metabolismo , Músculo Liso/metabolismo , Miosinas/metabolismo , Bexiga Urinária/metabolismo , Actinas/metabolismo , Medula Suprarrenal/metabolismo , Animais , ATPase de Ca(2+) e Mg(2+)/isolamento & purificação , Cálcio/farmacologia , Cromatografia em Gel , Cromatografia por Troca Iônica , Reações Cruzadas , Eletroforese em Gel de Poliacrilamida , Cinética , Substâncias Macromoleculares , Peso Molecular , Miosinas/química , Miosinas/isolamento & purificação , Suínos
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