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1.
Clin Otolaryngol ; 40(6): 646-50, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25858299

RESUMO

OBJECTIVES: Poor camera control during endoscopic dacryocystorhinostomy (EnDCR) surgery can cause inadequate visualisation of the anatomy and suboptimal surgical outcomes. This study investigates the feasibility of using computer vision tracking in EnDCR surgery as a potential formative feedback tool for the quality of endoscope control. DESIGN: A prospective cohort analysis was undertaken comparing junior versus senior surgeons performing routine EnDCR surgery. Computer vision tracking was applied to endoscopic video footage of the surgery: Total number of movements, camera path length in pixels and surgical time were determined for each procedure. A Mann-Whitney U-test was used to test for a significant difference between juniors and seniors (P < 0.05). SETTING: Operating theatre. PARTICIPANTS: Ten junior surgeons (<20 completed procedures) and 10 senior surgeons (>100 completed procedures). MAIN OUTCOME MEASURES: Total number of movements of the endoscope per procedure. Path length of the endoscope per procedure. RESULTS: Twenty videos, 10 from junior surgeons and 10 from senior surgeons were analysed. Feasibility of our tracking system was demonstrated. Mean camera path lengths were significantly different at 119,329px (juniors) versus 43,697px (seniors), P ≪ 0.05. The mean number of movements was significantly different at 9134 (juniors) versus 3690 (seniors), P ≪ 0.05. These quantifiable differences demonstrate construct validity for computer vision endoscope tracking as a measure of surgical experience. CONCLUSIONS: Computer vision tracking is a potentially useful structured and objective feedback tool to assist trainees in improving endoscope control. It enables juniors to examine how their pattern of endoscope control differs from that of seniors, focusing in particular on sections where they are most divergent.


Assuntos
Dacriocistorinostomia/métodos , Endoscopia/métodos , Doenças do Aparelho Lacrimal/cirurgia , Gravação em Vídeo/instrumentação , Adolescente , Adulto , Desenho de Equipamento , Feminino , Humanos , Masculino , Salas Cirúrgicas , Estudos Prospectivos , Reprodutibilidade dos Testes , Adulto Jovem
2.
Endoscopy ; 42(10): 790-9, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20886398

RESUMO

BACKGROUND AND STUDY AIMS: The impact of the diagnosis and treatment of dysplastic Barrett's esophagus on quality of life (QoL) is poorly understood. This study assessed the influence of dysplastic Barrett's esophagus on QoL and evaluated whether endoscopic treatment of dysplastic Barrett's esophagus with radiofrequency ablation (RFA) improves QoL. PATIENTS AND METHODS: We analyzed changes in QoL in the AIM Dysplasia Trial, a multicenter study of patients with dysplastic Barrett's esophagus who were randomly allocated to RFA therapy or a sham intervention. We developed a 10-item questionnaire to assess the influence of dysplastic Barrett's esophagus on QoL. The questionnaire was completed by patients at baseline and 12 months. RESULTS: 127 patients were randomized to RFA (n = 84) or sham (n = 43). At baseline, most patients reported worry about esophageal cancer (71 % RFA, 85 % sham) and esophagectomy (61 % RFA, 68 % sham). Patients also reported depression, impaired QoL, worry, stress, and dissatisfaction with the condition of their esophagus. Of those randomized, 117 patients completed the study to the 12-month end point. Compared with the sham group, patients treated with RFA had significantly less worry about esophageal cancer ( P=0.003) and esophagectomy ( P =0.009). They also had significantly reduced depression ( P=0.02), general worry about the condition of their esophagus ( P≤0.001), impact on daily QoL ( P=0.009), stress ( P=0.03), dissatisfaction with the condition of their esophagus ( P≤0.001), and impact on work and family life ( P=0.02). CONCLUSIONS: Inclusion in the treatment group of this randomized, sham-controlled trial of RFA was associated with improvement in disease-specific health-related quality of life. This improvement appears secondary to a perceived decrease in the risk of cancer.


Assuntos
Esôfago de Barrett/psicologia , Esôfago de Barrett/cirurgia , Ablação por Cateter , Qualidade de Vida/psicologia , Idoso , Ansiedade/etiologia , Distribuição de Qui-Quadrado , Neoplasias Esofágicas/prevenção & controle , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Lesões Pré-Cancerosas/prevenção & controle , Estatísticas não Paramétricas , Inquéritos e Questionários
3.
Parasite Immunol ; 32(4): 252-8, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20398225

RESUMO

Although there is an effective drug (praziquantel) available for the treatment of schistosomiasis, yet the disease is still spreading unabated and is rampant in 76 countries. Control via praziquantel treatment has so far been insufficient in reducing the disease transmission. Therefore, a vaccine in addition to other strategies, for example, improving sanitation and introduction of new drugs are essential to successfully control and eventually eradicate schistosomiasis. To this effect, we have targeted a functionally important antigen, Sm-p80 as a vaccine candidate. In this study, full length cDNA of Sm-p80 was cloned in VR1020, a FDA approved vector for human use. The protective efficacy of this vaccine formulation was tested in a murine model. Sm-p80-VR1020 vaccine formulation was able to induce 47% reduction in worm burden. Serology on samples obtained from vaccinated animals revealed a strong antibody response which included IgG and all of its subtypes, IgM and IgA. Proliferating splenocytes in response to recombinant Sm-p80 produced a wide spectrum of cytokines representing Th1, Th2 and Th17 types, as ascertained via RT-PCR analysis. These findings further strengthen the importance of Sm-p80 molecule as a vaccine candidate for intestinal schistosomiasis.


Assuntos
Antígenos de Helmintos/imunologia , Schistosoma mansoni/imunologia , Esquistossomose mansoni/prevenção & controle , Vacinas de DNA/imunologia , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/genética , Proliferação de Células , Citocinas/metabolismo , Feminino , Vetores Genéticos , Humanos , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Leucócitos Mononucleares/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Schistosoma mansoni/genética , Esquistossomose mansoni/imunologia , Baço/imunologia , Vacinas de DNA/genética
4.
Parasite Immunol ; 31(3): 156-61, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19222788

RESUMO

No effective vaccine exists for the human parasitic disease, schistosomiasis. We have targeted a functionally important antigen, Sm-p80 as a vaccine candidate because of its consistent immunogenicity, protective potential and important role in the immune evasion process. In this study we report that a Sm-p80-based DNA vaccine formulation confers 59% reduction in worm burden in mice. Animals immunized with Sm-p80-pcDNA3 exhibited a decrease in egg production by 84%. Sm-p80 DNA elicited strong immune responses that include IgG2A and IgG2B antibody isotypes in vaccinated animals. Splenocytes proliferated in response to Sm-p80 produced appreciably more Th1 response enhancing cytokines (IL-2, IFN-gamma) than Th2 response enhancing cytokines (IL-4, IL-10). These data reinforce the potential of Sm-p80 as an excellent vaccine candidate for schistosomiasis.


Assuntos
Antígenos de Helmintos/imunologia , Schistosoma mansoni/imunologia , Esquistossomose mansoni/prevenção & controle , Vacinas de DNA/imunologia , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/genética , Proliferação de Células , Citocinas/metabolismo , Feminino , Imunoglobulina G/sangue , Leucócitos Mononucleares/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Contagem de Ovos de Parasitas , Schistosoma mansoni/genética , Baço/imunologia , Vacinas de DNA/genética
5.
J Enzyme Inhib Med Chem ; 24(3): 876-82, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18951282

RESUMO

Several substituted phenoxy acetic acid derived pyrazolines were synthesized by the reaction between 2-{4-[3-(2,4-dihydroxyphenyl)-3-oxo-1-propenyl]-2-methoxyphenoxy} acetic acid and substituted acid hydrazides and were tested for their in vitro cytotoxicity and antiviral activity. None of the compounds showed any specific antiviral activity [50% antivirally effective concentration (EC(50)) > or = 5-fold lower than minimum cytotoxic concentration]. The most cytotoxic of the series was 2-{4-[3-(2,4-dihydroxyphenyl)-1-(2-hydroxybenzoyl-4,5-dihydro-1H-5-pyrazolyl]-2-methoxyphenoxy}acetic acid (3(j)), with a minimum cytotoxic concentration of 0.16 microg/mL in human embryonic lung (HEL) cells.


Assuntos
Acetatos/química , Acetatos/farmacologia , Antivirais/síntese química , Antivirais/farmacologia , Pirazóis/química , Pirazóis/farmacologia , Acetatos/toxicidade , Antivirais/química , Antivirais/toxicidade , Linhagem Celular , Humanos , Pulmão/citologia , Pulmão/embriologia , Testes de Sensibilidade Microbiana , Pirazóis/toxicidade , Relação Estrutura-Atividade
6.
Dis Esophagus ; 22(3): 216-22, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19207544

RESUMO

In patients undergoing chemoradiotherapy for esophageal cancer, the inability to eat may severely impair nutritional status. We conducted a retrospective study to compare the efficacy of the Polyflex self-expanding silicone stent (PS) versus a jejunostomy tube (JT) for maintaining nutrition during neoadjuvant chemoradiation therapy in patients with esophageal cancer who were scheduled for resectional surgery. Thirty-six patients were treated either with PS placement (12 patients) or JT placement (24 patients) prior to receiving an 8-week course of chemoradiation therapy. Patients were interviewed weekly until cessation of therapy. Patient data were collected on procedural success and complication rates, nutritional status, and dysphagia scores. PS placement was successful in 11 of 12 patients (92%), and those 11 patients were able to resume oral nutrition. Dysphagia scores improved from a mean of 3 to 1 in the PS group (P < 0.005) but did not change significantly in the JT group. PS were removed endoscopically without complications prior to the esophagectomies. Albumin levels and weight increased significantly in both the PS and JT groups. There were no significant differences between groups in the procedural success rates (PS 92% vs. JT 100%, P = 0.33), complication rates (PS 22% vs. JT 4%, P = 0.11), mean increase in weight (PS 4.4 kg vs. JT 4.2 kg, P = 0.59), and mean increase in serum albumin (PS 0.62 g/dL vs. JT 0.44 g/dL, P = 0.05). PS is a safe and effective alternative to a surgical JT for maintaining nutrition in this subset of patients.


Assuntos
Transtornos de Deglutição/terapia , Nutrição Enteral , Neoplasias Esofágicas/complicações , Jejunostomia/instrumentação , Stents , Adenocarcinoma/complicações , Adenocarcinoma/terapia , Idoso , Idoso de 80 Anos ou mais , Peso Corporal , Carcinoma de Células Escamosas/complicações , Carcinoma de Células Escamosas/terapia , Estudos de Casos e Controles , Transtornos de Deglutição/etiologia , Endoscopia Gastrointestinal , Neoplasias Esofágicas/terapia , Estenose Esofágica/etiologia , Estenose Esofágica/terapia , Feminino , Humanos , Laparoscopia , Masculino , Pessoa de Meia-Idade , Terapia Neoadjuvante , Estado Nutricional , Estudos Retrospectivos , Albumina Sérica/análise , Índice de Gravidade de Doença
7.
Eur J Med Chem ; 43(11): 2331-7, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18031870

RESUMO

A series of novel 1-substituted-4-(2-methylphenyl)-4H-[1,2,4]triazolo[4,3-a]quinazolin-5-ones were synthesized by the cyclization of 2-hydrazino-3-(2-methylphenyl)-3H-quinazolin-4-one with various one carbon donors. The starting material 2-hydrazino-3-(2-methylphenyl)-3H-quinazolin-4-one was synthesized from 2-methyl aniline by a novel innovative route. The title compounds were tested for their in vivo H(1)-antihistaminic activity on guinea pigs; all the tested compounds protected the animals from histamine-induced bronchospasm significantly. Compound 1-methyl-4-(2-methylphenyl)-4H-[1,2,4]triazolo[4,3-a]quinazolin-5-one (II) emerged as the most active compound of the series and it is more potent (72.45%) when compared to the reference standard chlorpheniramine maleate (71%). Compound II showed negligible sedation (11%) when compared to chlorpheniramine maleate (30%). Hence it could serve as the prototype molecule for further development as a new class of H(1)-antihistaminic agents.


Assuntos
Antagonistas dos Receptores Histamínicos H1/síntese química , Antagonistas dos Receptores Histamínicos H1/farmacologia , Quinazolinas/síntese química , Quinazolinas/farmacologia , Triazóis/síntese química , Triazóis/farmacologia , Animais , Sistema Nervoso Central/efeitos dos fármacos , Cobaias , Antagonistas dos Receptores Histamínicos H1/química , Antagonistas dos Receptores Histamínicos H1/classificação , Masculino , Metilação , Estrutura Molecular , Quinazolinas/química , Quinazolinas/classificação , Relação Estrutura-Atividade , Triazóis/química , Triazóis/classificação
8.
Diabetes Metab Syndr ; 12(6): 839-842, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28899639

RESUMO

BACKGROUND: It is being increasingly reported that some of the youth onset diabetes patients cannot be classified clearly as type 1 diabetes mellitus (T1DM) or type 2 diabetes mellitus (T2DM) based on usual criteria and the term double diabetes (DD) coined for these cases. AIM: The objective of the study was to find out the prevalence of DD in youth onset diabetes patients from east Delhi and neighboring NCR region. METHODS: A total of 200 patients with youth onset diabetes below 25 years of age were recruited from a tertiary care hospital in East Delhi. Clinical history, family history of diabetes and anthropometry of patients were recorded. Fasting serum C-peptide, Anti-IA2-antibody and Anti-GAD-antibody were measured in all patients. Patients positive for Anti-GAD-antibody (>1.05U/ml) and C-peptide level >0.3nmol/l were characterized as DD patients. Patients negative for Anti-GAD-antibody and C-peptide >0.3nmol/l were kept under the category of T2DM. Patients with low C-peptide level along with one of the following, positive Anti-GAD-antibody, positive Anti-IA2-antibody and diabetic ketoacidosis (DKA) were considered as T1DM. Remaining patients were kept under the unknown category. RESULTS: Mean age of study subjects was 18.2±7.1years. Seven percent (7%) of the subjects were classified as DD, 51% as T1DM, 13% as T2DM and 29% were kept under the unknown category. Mean age of subjects with 22.2±9.7, 16.9±6.7, 20.6±7.7 and 19.4±7.4 years in DD, T1DM, T2DM and unknown category respectively. Mean BMI of subjects with DD, T1DM, T2DM and unknown category was 19.8±5.7, 16.6±3.7, 19.3±4.1 and 18.0±4.6 kg/m2 respectively. CONCLUSION: Double diabetes is an important occurrence among youth onset diabetes subjects. Only half of the subjects with youth onset of diabetes had T1DM.


Assuntos
Diabetes Mellitus Tipo 1/epidemiologia , Diabetes Mellitus Tipo 2/epidemiologia , Adolescente , Idade de Início , Criança , Feminino , Humanos , Índia/epidemiologia , Masculino , Prevalência , Adulto Jovem
9.
Diabetes Metab Syndr ; 12(3): 313-316, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29287842

RESUMO

AIM: To find out the prevalence of obesity and glucose intolerance among nurses working in tertiary care hospital. METHODS: Study was conducted in 496 apparently healthy females comprising two groups. Group B had 290 nurses and control group A had 206 age matched female subjects of general population. Detailed performa was filled which included anthropometry, systemic examination and other details. Fasting plasma glucose was done followed by oral glucose tolerance test (OGTT). Subjects with body mass index ≥23 kg/m2 were categorized as 'overweight' and ≥25 kg/m2 as 'obese' as per criteria for Asian Indians. Women with waist circumference of ≥80 cm were categorized as 'centrally obese'. RESULTS: Mean age of subjects in groups A and B was 40.45 ±â€¯8.64 years and 40.50 ±â€¯6.96 years respectively. Significantly higher number of nurses (80%) were overweight or obese compared to controls (59.71%,P = < .001). Similarly, central obesity was significantly higher in nurses (82.07%) compared to controls (67.96%,P = <.001). The prevalence of glucose intolerance (prediabetes and newly detected diabetes) was significantly higher in controls compared to nurses (45.63% vs 29.66%, P < .001). CONCLUSION: Every four out of five nurses working in tertiary care hospital have overweight/obesity and central obesity. Despite this they have lower rates of glucose intolerance.


Assuntos
Biomarcadores/análise , Intolerância à Glucose , Obesidade/complicações , Obesidade/epidemiologia , Estado Pré-Diabético/etiologia , Centros de Atenção Terciária , Adulto , Antropometria , Glicemia/análise , Índice de Massa Corporal , Estudos de Casos e Controles , Estudos Transversais , Feminino , Seguimentos , Teste de Tolerância a Glucose , Hemoglobinas Glicadas/análise , Humanos , Índia/epidemiologia , Enfermeiras e Enfermeiros , Prevalência , Prognóstico , Fatores de Risco
10.
Aliment Pharmacol Ther ; 24(11-12): 1623-30, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17206950

RESUMO

AIM: To determine factors affecting compliance of a follow-up colonoscopy in patients with previously diagnosed adenomatous colon polyps. METHODS: A retrospective review was performed on patients with adenomatous polyps excised between January and December 1998. Twenty-nine clinical factors were assessed in patients grouped into whether they were compliant (n = 81) or noncompliant (n = 38) with follow-up colonoscopy. Significant variables by univariate analysis were included in multivariate regression. RESULTS: One hundred and nineteen patients with adenomatous colon polyps were identified. Of 119 patients, 114 had a documented recommendation for follow-up of 5 years or less, with 69% having been compliant. In a univariate analysis, greater number of polyps (P = 0.04), NSAID use (P = 0.02), statin use (P = 0.005), first-degree relatives with colon cancer (P = 0.05) and compliance with out-patient clinic follow-up (P < 0.001) were significantly associated with patient compliance. Multivariate analysis revealed statin use (P = 0.05), first-degree relatives with colon cancer (P = 0.06) and compliance with out-patient clinic follow-up (P < 0.001) were independent predictors of compliance. CONCLUSIONS: History of statin use and family history of colon cancer are good predictors of compliance. The strongest predictor can be anticipated with compliance assessed with encounters for other visits. Strong efforts should be directed at improving patient education about colon cancer by the physician and facilitating patient compliance.


Assuntos
Pólipos Adenomatosos/patologia , Neoplasias do Colo/patologia , Pólipos do Colo/patologia , Colonoscopia , Cooperação do Paciente , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
11.
Aliment Pharmacol Ther ; 24(7): 1059-66, 2006 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-16984500

RESUMO

BACKGROUND: Laparoscopic cholecystectomy (LC) is recommended for patients with choledocholithiasis after ERCP with sphincterotomy (ES) and stone extraction. AIM: We designed a decision model to address whether ES alone versus ES followed by LC (ES + LC) is the optimal treatment in high-risk patients with choledocholithiasis. METHODS: Our cohort were patients with obstructive jaundice who have undergone an ES with biliary clearance. Recurrent biliary complications over a 2-year period stratified by gallbladder status (in/out) and age-stratified surgical complication rates were obtained from the literature. Failure of therapy was defined as either recurrent symptoms or death attributed to biliary complications. RESULTS: For age 70-79 years, ES failed in 15% whereas ES + LC failed in 17% of cases. Mortality in the EC + LC group was 3.4 times that of the ES alone cohort. For age 80+ years, ES was dominant with an incremental success rate of 8%. Mortality in the ES + LC was 7.6 times that of ES. For age <70, ES + LC was the dominant strategy with an incremental success rate 5%. Sensitivity analysis in the groups confirmed our conclusions. CONCLUSIONS: Management of choledocholithiasis by ES and stone clearance, but without cholecystectomy, should be considered for patients aged 70+. For low-risk patients, ES + LC should be performed to prevent recurrent biliary complications.


Assuntos
Colecistectomia/métodos , Coledocolitíase/cirurgia , Esfinterotomia Endoscópica/métodos , Idoso , Idoso de 80 Anos ou mais , Estudos de Coortes , Técnicas de Apoio para a Decisão , Humanos , Fatores de Risco , Taxa de Sobrevida , Resultado do Tratamento
12.
Prog Mol Biol Transl Sci ; 142: 291-315, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27571699

RESUMO

There is an urgent need to develop new vaccines for tuberculosis, HIV/AIDS, and malaria, as well as for chronic and debilitating infections known as neglected tropical diseases (NTDs). The term "NTD" emerged at the beginning of the new millennium to describe a set of diseases that are characterized as (1) poverty related, (2) endemic to the tropics and subtropics, (3) lacking public health attention and inadequate industrial investment, (4) having poor research funding and a weak research and development (R&D) pipeline, (5) usually associated with high morbidity but low mortality, and (6) often having no safe and long-lasting treatment available. Many additional challenges to the current control and elimination programs for NTDs exist. These include inconsistent performance of diagnostic tests, regional differences in access to treatment and in treatment outcome, lack of integrated surveillance and vector/intermediate host control, and impact of ecological climatic changes particularly in regions where new cases are increasing in previously nonendemic areas. Moreover, the development of NTD vaccines, including those for schistosomiasis, leishmaniasis, leprosy, hookworm, and Chagas disease are being led by nonprofit product development partnerships (PDPs) working in partnership with academic and industrial partners, contract research organizations, and in some instances vaccine manufacturers in developing countries. In this review, we emphasize global efforts to fuel the development of NTD vaccines, the translational activities needed to effectively move promising vaccine candidates to Phase-I clinical trials and some of the hurdles to ensuring their availability to people in the poorest countries of Africa, Asia, Latin America, and the Caribbean.


Assuntos
Doenças Negligenciadas/terapia , Pesquisa Translacional Biomédica , Medicina Tropical , Antígenos/metabolismo , Doenças Negligenciadas/economia , Medicina Tropical/economia , Vacinas/imunologia
13.
Trop Biomed ; 33(4): 652-662, 2016 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-33579061

RESUMO

The current epidemiological study was designed to trace the involved risk factors in Hepatitis C Virus (HCV) spread and to identify any association between HCV genotypes and risk factors. Blood samples were taken from 400 participants and viral genotyping was performed in order to find any possible relationship between the risk factors and genotypes. Major genotypes included 3, 1, 4 and several untypeable ones with prevalence rates 65%, 22.5%, 2.75% and 9.75% respectively. Surgery and dental procedure were strongly related to the spread of genotype 3b, while genotype 1b was strongly related to blood transfusion and dental procedures as a single combination risk factor. On the other hand genotypes 1a, 3a, 4 and the untypeable genotypes, were equally affected by all reported risk factors. The probability of occurrence of genotype 3a with reference to dental procedures was 11%. Dental procedures, unsafe injection and surgical procedures are the main risk factors while the blood transfusion in combination with dental procedures has emerged as a potent risk factor in the transmission of HCV.

14.
Biochim Biophys Acta ; 1181(1): 37-44, 1993 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-8457603

RESUMO

Calcium-dependent, neutral cysteine-proteases (calpain) were purified from human blood flukes, Schistosoma mansoni. The electrophoretic mobilities, Western blot analyses and high specificity to peptide inhibitors confirmed the presence of both calpain I and II in the purified preparation. The schistosome calpains were localized in the surface syncytial epithelium and underlying musculature. Using peptide inhibitors, calpain was shown to function as a mediator of the surface membrane synthetic process. Since there was also no immunological cross-reactivity between vertebrate and schistosome calpains using antibodies affinity-purified from native and recombinant schistosome calpains, this protease may be usefully investigated as forming the basis of a molecular vaccine against schistosomiasis.


Assuntos
Calpaína/isolamento & purificação , Schistosoma mansoni/química , Animais , Western Blotting , Calpaína/antagonistas & inibidores , Membrana Celular/metabolismo , Colina/metabolismo , Cricetinae , Reações Cruzadas , Endopeptidases/metabolismo , Cinética , Mesocricetus , Metionina/metabolismo , Microscopia Eletrônica de Varredura , Schistosoma mansoni/metabolismo , Schistosoma mansoni/ultraestrutura
15.
Parasite ; 12(1): 3-8, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15828575

RESUMO

Despite advances in control via snail eradication and large-scale chemotherapy using praziquental, schistosomiasis continues to spread to new geographic areas particularly in sub-Saharan Africa. Presently, there is no vaccine for controlling this disease. We have concentrated on a functionally important schistosome antigen Sm-p80 as a possible vaccine candidate for schistosomiasis. Here we report the proliferation of spleen cells in response to the recombinant Sm-p80 protein and cytokine (IFN-gamma and IL-4) production by the splenocytes. These spleen cells were obtained from groups of mice that were vaccinated with a DNA vaccine formulation containing Sm-p80 and one of the Th-1 (IL-2 or IL-12) or Th-2 (GM-CSF, IL-4) enhancer cytokines. The splenocytes from the groups of mice vaccinated with Sm-p80 DNA in the presence of Th-2 enhancer cytokines showed moderate but detectable proliferation. The splenocytes obtained from mice vaccinated with Sm-p80 DNA with Th-1 enhancer cytokines IL-2 and IL-12 provided the highest proliferation. The IFN-gamma production by splenocytes was found to follow the similar pattern [(Sm-p80) < (Sm-p80 + IL-4) < (Sm-p80 + GMCSF) < (Sm-p80 + IL-12) < (Sm-p80 + IL-2)], as has been observed for the proliferation and protection data. However, the elevated IL-4 production was inversely correlated to Sm-p80-induced splenocyte proliferation or the protection. These results show again that protective immune response induced by Sm-p80 is of Th-1 type.


Assuntos
Adjuvantes Imunológicos/farmacologia , Calpaína/imunologia , Schistosoma mansoni/imunologia , Esquistossomose mansoni/prevenção & controle , Baço/citologia , Vacinas de DNA/imunologia , Animais , Citocinas/imunologia , Imunização , Interferon gama/biossíntese , Interferon gama/farmacologia , Interleucina-4/farmacologia , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Subunidades Proteicas , Esquistossomose mansoni/imunologia , Baço/imunologia
16.
J Interferon Cytokine Res ; 19(8): 923-8, 1999 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10476939

RESUMO

ISG-15 is a 15-kDa protein encoded by an interferon (IFN)-stimulated gene (ISG), which is transcriptionally regulated by IFN-alpha and IFN-beta. Considered as part of the cytokine network, ISG-15 has the potential to amplify the immunomodulatory effects of these IFNs by enhancing IFN-gamma production, natural killer cell proliferation, and lymphokine-alphactivated killer cell cytotoxicity. To understand better the mechanism(s) of action of orally administered IFN-alpha, we have studied the effect of IFN-alpha on ISG-15 gene expression by human buccal epithelial cells (BEC). For in vitro studies, ISG-15 mRNA and protein levels were measured in BEC incubated for 0.5, 2, and 9 h with 100 or 1,000 IU/ml of human lymphoblastoid IFN-alpha. For in vivo studies, ISG-15 mRNA was measured in BEC samples collected at baseline, and 0.5, 2, and 9 h after 5-20 min of oral rinsing with 10 ml of IFN-alpha (1,000 IU/ml). ISG-15 mRNA was measured by reverse transcriptase polymerase chain reaction (RT-PCR), and ISG-15 protein production by Western Blot analysis. IFN-alpha augmented BEC ISG-15 gene expression in a concentration dependent manner both in vivo and in vitro. We conclude that orally administered IFN-alpha exerts its immunomodulatory effects in humans in part by upregulating the production of ISG-15 by BEC, thereby enhancing the immune reactivity of mucosa-associated lymphocytes.


Assuntos
Adjuvantes Imunológicos/uso terapêutico , Citocinas/genética , Células Epiteliais/efeitos dos fármacos , Interferon-alfa/uso terapêutico , Mucosa Bucal/efeitos dos fármacos , Transcrição Gênica , Ubiquitinas/análogos & derivados , Administração Oral , Divisão Celular/efeitos dos fármacos , Bochecha , Humanos , Técnicas In Vitro , Células Matadoras Naturais/citologia , Células Matadoras Naturais/efeitos dos fármacos , Peso Molecular , Mucosa Bucal/citologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estimulação Química
17.
J Interferon Cytokine Res ; 19(8): 929-35, 1999 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10476940

RESUMO

Aquaporins are a family of homologous membrane proteins that function as highly selective water channels. Aquaporin-5 (AQP5) is uniquely present in lacrimal and salivary glands, where it accounts for normal tear and saliva production. We tested the hypothesis that orally administered human interferon-alpha (HuIFN-alpha) benefits persons with xerostomia by augmenting the production of AQP5 protein by parotid gland epithelium. Cells from three human parotid glands were cultured with and without human lymphoblastoid IFN-alpha, and assayed for AQP5 mRNA levels by reverse transcriptase polymerase chain reaction (RT-PCR), and AQP5 protein levels by Western blot. Intracellular localization of AQP5 protein was done using confocal microscopy. The functional integrity of the glandular tissue was confirmed by RT-PCR analysis of alpha-amylase 1 and basic proline-rich protein transcripts. AQP5 was constitutively expressed in human parotid gland tissue, with AQP5 protein restricted to the plasma membranes and cytoplasmic vesicles of acinar cells. IFN-alpha augmented AQP5 transcription and protein production in a concentration-dependent manner, and increased the size of intensity of staining of AQP5-containing cytoplasmic vesicles in acinar cells. We conclude that IFN-alpha upregulates AQP5 gene expression in human parotid acinar cells in vitro. To our knowledge, this is the first demonstration that IFN-alpha regulates the gene expression of an aquaporin.


Assuntos
Aquaporinas/genética , Interferon-alfa/uso terapêutico , Proteínas de Membrana , Glândula Parótida/efeitos dos fármacos , Administração Oral , Aquaporina 5 , Células Cultivadas , Avaliação Pré-Clínica de Medicamentos , Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , Glândula Parótida/citologia , Glândula Parótida/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para Cima
18.
Mol Biochem Parasitol ; 25(1): 19-28, 1987 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-2444880

RESUMO

Gene expression and its regulation was studied in the rat tapeworm, Hymenolepis diminuta, during strobilization. RNAs extracted from the different developmental stages of the parasite were translated in vitro in a rabbit reticulocyte lysate system. Two-dimensional patterns of the translational products were compared with the 2-dimensional patterns obtained by metabolic labelling with [35S]methionine. The results indicated a post-transcriptional regulation of gene expression during strobilization in this parasite. Gene expression and its regulation was also studied in H. diminuta obtained from 'non-permissive' hosts (mice) and immune suppressed mice and compared with the parasites of the same age obtained from 'permissive' rats hosts. The 2-dimensional patterns of the in vitro translation products, obtained by translating the RNA of different groups of parasites, were compared with the patterns of gene products obtained from parasites using [35S]methionine metabolic labelling. The results indicated a massive post-transcriptional regulation of gene expression, the latter inhibited in parasites obtained from normal mice, but not in immune suppressed mice.


Assuntos
Regulação da Expressão Gênica , Hymenolepis/genética , Animais , Eletroforese em Gel de Poliacrilamida , Feminino , Hymenolepis/crescimento & desenvolvimento , Hymenolepis/imunologia , Tolerância Imunológica , Masculino , Camundongos , Fotofluorografia , Biossíntese de Proteínas , Proteínas/análise , Proteínas/genética , RNA/genética , RNA Mensageiro/genética , Ratos
19.
Mol Biochem Parasitol ; 40(1): 95-103, 1990 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-2348834

RESUMO

A procedure is described whereby a highly enriched plasma membrane fraction was isolated from Trypanosoma brucei by the technique of preparative free-flow electrophoresis. The purity of the plasma membrane fraction was monitored by electron microscopy and by marker enzymology, and is compared to those obtained by previous methods. Proteins associated with plasma membrane fraction were analyzed by SDS-PAGE and phase separated in Triton X-114.


Assuntos
Membrana Celular/ultraestrutura , Trypanosoma brucei brucei/ultraestrutura , Animais , Fracionamento Celular , Membrana Celular/enzimologia , Eletroforese , Microscopia Eletrônica
20.
Mol Vis ; 6: 144-7, 2000 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-10930475

RESUMO

The present study focuses on the detection of differentially expressed genes in migrating (healing) and nonmigrating (normal) corneal epithelium on agarose gel using a modified procedure of differential display reverse transcriptase-polymerase chain reaction (DDRT-PCR). Rabbit corneal epithelial organ cultures were used to obtain nonmigrating and migrating samples. RNA was extracted using Trizol LS reagent. PCR was modified in order to allow detection of amplified products on 3% agarose gel with ethidium bromide staining. Products were also resolved on 6% denaturing polyacrylamide-urea gels and observed by silver staining. Agarose gels showed two prominent bands that were heavily expressed in the 458 bp and 587 bp region of the nonmigrating samples. In addition light bands were visible in the region corresponding to 234 bp and 450 bp. In the migrating samples, two light bands were visible in the region of 267 bp and 300 bp. Eight amplicons, six in the nonmigrating corneal epithelial sample and two in the migrating corneal epithelial samples, were also found to be differentially expressed when products were run on 6% denaturing polyacrylamide-urea gels. Thus, DDRT-PCR products can be detected on agarose gels and prove very helpful and economical in the initial studies of DDRT-PCR.


Assuntos
Epitélio Corneano/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Animais , Técnicas de Cultura , Eletroforese em Gel de Ágar , Eletroforese em Gel de Poliacrilamida , RNA Mensageiro/análise , Coelhos , Coloração pela Prata , Cicatrização
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