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1.
Iran J Basic Med Sci ; 23(3): 396-405, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32440328

RESUMO

OBJECTIVES: The role of N-acetylcysteine (NAC) as an anti-oxidant in attenuating bleomycin-induced pulmonary fibrosis has been reported. However, its effect on parenchymal remodeling via regulating the protease-antiprotease balance is not fully defined. Therefore, the present study was designed to explore the possible role of matrix metalloproteinases (MMP), tissue inhibitors of metalloproteinases (TIMP) and transforming growth factor-ß1 (TGF-ß1) pathway and their modulation by NAC in attenuating bleomycin-induced pulmonary fibrosis in rats. MATERIALS AND METHODS: Bleomycin sulphate (7 units/kg) was instilled inside the trachea to induce pulmonary fibrosis. The time course of TGF-ß1, MMP-9, TIMP-1,3 mRNA and protein expression, TGF-ß1 and hydroxyproline levels were evaluated on days 7, 14, and 28. NAC (0.3 mmol/kg and 3 mmol/kg) was administered in bleomycin-instilled animals. RESULTS: NAC treatment significantly attenuated bleomycin-induced histopathological changes by decreasing interstitial inflammation and reducing the deposition of extracellular matrix proteins such as collagen. Moreover, it increased the mRNA and protein expression of MMP-9 and decreased the expression of TIMP-1,3 in alveolar epithelial cells (AECs), interstitial macrophages and inflammatory cells. Indeed, there was decrease in the MMP-9/TIMP ratio in bleomycin-instilled rats, which increased with NAC treatment. Moreover, NAC attenuated bleomycin-induced increased expression of TGF-ß1 and total lung collagen levels. CONCLUSION: NAC attenuates bleomycin-induced pulmonary fibrosis by normalizing the protease-antiprotease balance and favoring the degradation of collegen to reduce fibrosis.

2.
COPD ; 15(6): 559-571, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30663435

RESUMO

Adenosine is a signaling molecule which is produced in high concentrations during airway inflammation. Airway inflammation is a characteristic feature of COPD. Therefore, the current study was designed to evaluate the changes in adenosine metabolism in COPD and correlate these changes with severity of the disease. The study was conducted on 50 healthy controls (25 healthy non-smokers and 25 healthy smokers) and 46 COPD patients (21 moderate, 15 severe and 10 very severe). The patients were sub-divided into moderate, severe and very severe categories as per the GOLD spirometric classification. Blood was collected from each subject and serum, lymphocytes and erythrocytes were separated. The adenosine levels and activities of 5'-nucleotidase, adenosine deaminase and its isoenzymes were assessed in serum, lymphocytes and erythrocytes. The data were analyzed statistically. A p value < 0.05 was considered as significant. In healthy smokers and COPD patients the adenosine levels increased. In COPD patients 5'-nucleotidase activity increased significantly in serum, lymphocytes and erythrocytes. The activities of ADA and isoenzymes decreased significantly in serum of healthy smokers and COPD patients, in lymphocytes and erythrocytes of very severe COPD patients and of ADA and ADA2 in lymphocytes and erythrocytes of moderate and severe COPD patients. The FEV1 (% of predicted) showed a significant negative correlation with adenosine levels and 5'-nucleotidase activity in serum, lymphocytes and erythrocytes and significant positive correlation with ADA and isoenzymes activity in serum and lymphocytes of COPD patients. We conclude that the adenosine metabolism changes in COPD. The adenosine levels and 5'-nucleotidase activity increase, and ADA activity decreases with severity of the disease.


Assuntos
5'-Nucleotidase/sangue , Adenosina Desaminase/sangue , Adenosina/sangue , Doença Pulmonar Obstrutiva Crônica/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Eritrócitos/metabolismo , Feminino , Volume Expiratório Forçado , Voluntários Saudáveis , Humanos , Isoenzimas/sangue , Linfócitos/metabolismo , Masculino , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Índice de Gravidade de Doença , Fumar/sangue
3.
Anal Chem ; 90(3): 1870-1880, 2018 02 06.
Artigo em Inglês | MEDLINE | ID: mdl-29276835

RESUMO

For LC-MS-based targeted quantification of biotherapeutics and biomarkers in clinical and pharmaceutical environments, high sensitivity, high throughput, and excellent robustness are all essential but remain challenging. For example, though nano-LC-MS has been employed to enhance analytical sensitivity, it falls short because of its low loading capacity, poor throughput, and low operational robustness. Furthermore, high chemical noise in protein bioanalysis typically limits the sensitivity. Here we describe a novel trapping-micro-LC-MS (T-µLC-MS) strategy for targeted protein bioanalysis, which achieves high sensitivity with exceptional robustness and high throughput. A rapid, high-capacity trapping of biological samples is followed by µLC-MS analysis; dynamic sample trapping and cleanup are performed using pH, column chemistry, and fluid mechanics separate from the µLC-MS analysis, enabling orthogonality, which contributes to the reduction of chemical noise and thus results in improved sensitivity. Typically, the selective-trapping and -delivery approach strategically removes >85% of the matrix peptides and detrimental components, markedly enhancing sensitivity, throughput, and operational robustness, and narrow-window-isolation selected-reaction monitoring further improves the signal-to-noise ratio. In addition, unique LC-hardware setups and flow approaches eliminate gradient shock and achieve effective peak compression, enabling highly sensitive analyses of plasma or tissue samples without band broadening. In this study, the quantification of 10 biotherapeutics and biomarkers in plasma and tissues was employed for method development. As observed, a significant sensitivity gain (up to 25-fold) compared with that of conventional LC-MS was achieved, although the average run time was only 8 min/sample. No appreciable peak deterioration or loss of sensitivity was observed after >1500 injections of tissue and plasma samples. The developed method enabled, for the first time, ultrasensitive LC-MS quantification of low levels of a monoclonal antibody and antigen in a tumor and cardiac troponin I in plasma after brief cardiac ischemia. This strategy is valuable when highly sensitive protein quantification in large sample sets is required, as is often the case in typical biomarker validation and pharmaceutical investigations of antibody therapeutics.


Assuntos
Cromatografia Líquida/instrumentação , Ensaios de Triagem em Larga Escala/instrumentação , Espectrometria de Massas/instrumentação , Peptídeos/análise , Proteínas/análise , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/análise , Biomarcadores/análise , Cromatografia Líquida/economia , Cromatografia Líquida/métodos , Desenho de Equipamento , Ensaios de Triagem em Larga Escala/economia , Ensaios de Triagem em Larga Escala/métodos , Humanos , Imunoglobulina G/análise , Limite de Detecção , Espectrometria de Massas/economia , Espectrometria de Massas/métodos , Camundongos , Ratos , Suínos
4.
Proteomics Insights ; 8: 1178641817700880, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28469466

RESUMO

The 2-dimensional gel electrophoresis (2-DE) technique is widely used for the analysis of complex protein mixtures extracted from biological samples. It is one of the most commonly used analytical techniques in proteomics to study qualitative and quantitative protein changes between different states of a cell or an organism (eg, healthy and diseased), conditionally expressed proteins, posttranslational modifications, and so on. The 2-DE technique is used for its unparalleled ability to separate thousands of proteins simultaneously. The resolution of the proteins by 2-DE largely depends on the quality of sample prepared during protein extraction which increases results in terms of reproducibility and minimizes protein modifications that may result in artifactual spots on 2-DE gels. The buffer used for the extraction and solubilization of proteins influences the quality and reproducibility of the resolution of proteins on 2-DE gel. The purification by cleanup kit is another powerful process to prevent horizontal streaking which occurs during isoelectric focusing due to the presence of contaminants such as salts, lipids, nucleic acids, and detergents. Erythrocyte membrane proteins serve as prototypes for multifunctional proteins in various erythroid and nonerythroid cells. In this study, we therefore optimized the selected major conditions of 2-DE for resolving various proteins of human erythrocyte membrane. The modification included the optimization of conditions for sample preparation, cleanup of protein sample, isoelectric focusing, equilibration, and storage of immobilized pH gradient strips, which were further carefully examined to achieve optimum conditions for improving the quality of protein spots on 2-DE gels. The present improved 2-DE analysis method enabled better detection of protein spots with higher quality and reproducibility. Therefore, the conditions established in this study may be used for the 2-DE analysis of erythrocyte membrane proteins for different diseases, which may help to identify the proteins that may serve as markers for diagnostics as well as targets for development of new therapeutic potential.

5.
Bioanalysis ; 8(14): 1465-79, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27329641

RESUMO

BACKGROUND: Alectinib is a novel anaplastic lymphoma kinase (ALK) inhibitor for treatment of patients with ALK-positive non-small-cell lung cancer who have progressed on or are intolerant to crizotinib. To support clinical development, concentrations of alectinib and metabolite M4 were determined in plasma from patients and healthy subjects. METHODS: LC-MS/MS methods were developed and validated in two different laboratories: Chugai used separate assays for alectinib and M4 in a pivotal Phase I/II study while Roche established a simultaneous assay for both analytes for another pivotal study and all other studies. CONCLUSION: Cross-validation assessment revealed a bias between the two bioanalytical laboratories, which was confirmed with the clinical PK data between both pivotal studies using the different bioanalytical methods.


Assuntos
Carbazóis/sangue , Carbazóis/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Piperidinas/sangue , Piperidinas/metabolismo , Inibidores de Proteínas Quinases/sangue , Inibidores de Proteínas Quinases/metabolismo , Receptores Proteína Tirosina Quinases/antagonistas & inibidores , Espectrometria de Massas em Tandem/métodos , Quinase do Linfoma Anaplásico , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/enzimologia , Cromatografia Líquida de Alta Pressão/instrumentação , Desenho de Equipamento , Humanos , Limite de Detecção , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/enzimologia , Espectrometria de Massas em Tandem/instrumentação
6.
Bioanalysis ; 6(24): 3355-68, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25534792

RESUMO

The 2014 8th Workshop on Recent Issues in Bioanalysis (8th WRIB), a 5-day full immersion in the evolving field of bioanalysis, took place in Universal City, California, USA. Close to 500 professionals from pharmaceutical and biopharmaceutical companies, contract research organizations and regulatory agencies worldwide convened to share, review, discuss and agree on approaches to address current issues of interest in bioanalysis. The topics covered included both small and large molecules, and involved LCMS, hybrid LBA/LCMS, LBA approaches and immunogenicity. From the prolific discussions held during the workshop, specific recommendations are presented in this 2014 White Paper. As with the previous years' editions, this paper acts as a practical tool to help the bioanalytical community continue advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2014 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 3) covers the recommendations for Large molecules bioanalysis using LBA and Immunogenicity. Part 1 (Small molecules bioanalysis using LCMS) and Part 2 (Hybrid LBA/LCMS, Electronic Laboratory Notebook and Regulatory Agencies' Input) were published in the Bioanalysis issues 6(22) and 6(23), respectively.


Assuntos
Técnicas de Química Analítica , Imunidade , Anticorpos Neutralizantes/imunologia , Biotransformação , Humanos , Preparações Farmacêuticas/metabolismo , Farmacocinética , Polietileno/química , Guias de Prática Clínica como Assunto , Estados Unidos , United States Food and Drug Administration
7.
Bioanalysis ; 6(23): 3237-49, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25529890

RESUMO

The 2014 8th Workshop on Recent Issues in Bioanalysis (8th WRIB), a 5-day full immersion in the evolving field of bioanalysis, took place in Universal City, California, USA. Close to 500 professionals from pharmaceutical and biopharmaceutical companies, contract research organizations and regulatory agencies worldwide convened to share, review, discuss and agree on approaches to address current issues of interest in bioanalysis. The topics covered included both small and large molecules, and involved LCMS, hybrid LBA/LCMS, LBA approaches and immunogenicity. From the prolific discussions held during the workshop, specific recommendations are presented in this 2014 White Paper. As with the previous years' editions, this paper acts as a practical tool to help the bioanalytical community continue advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2014 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 2) covers the recommendations for Hybrid LBA/LCMS, Electronic Laboratory Notebook and Regulatory Agencies' Input. Part 1 (Small molecules bioanalysis using LCMS) was published in the Bioanalysis issue 6(22) and Part 3 (Large molecules bioanalysis using LBA and Immunogenicity) will be published in the Bioanalysis issue 6(24).


Assuntos
Técnicas de Laboratório Clínico , Métodos Analíticos de Preparação de Amostras , Cromatografia Líquida , Humanos , Espectrometria de Massas
8.
Bioanalysis ; 6(22): 3039-49, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25496256

RESUMO

The 2014 8th Workshop on Recent Issues in Bioanalysis (8th WRIB), a 5-day full immersion in the evolving field of bioanalysis, took place in Universal City, California, USA. Close to 500 professionals from pharmaceutical and biopharmaceutical companies, contract research organizations and regulatory agencies worldwide convened to share, review, discuss and agree on approaches to address current issues of interest in bioanalysis. The topics covered included both small and large molecules, and involved LCMS, hybrid LBA/LCMS, LBA approaches and immunogenicity. From the prolific discussions held during the workshop, specific recommendations are presented in this 2014 White Paper. As with the previous years' editions, this paper acts as a practical tool to help the bioanalytical community continue advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2014 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 1) covers the recommendations for small molecule bioanalysis using LCMS. Part 2 (Hybrid LBA/LCMS, Electronic Laboratory Notebook and Regulatory Agencies' input) and Part 3 (Large molecules bioanalysis using LBA and Immunogenicity) will be published in the upcoming issues of Bioanalysis.


Assuntos
Bioensaio , Cromatografia Líquida/métodos , Espectrometria de Massas/métodos
10.
Bioanalysis ; 5(23): 2903-18, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24107004

RESUMO

The 2013 7th Workshop on Recent Issues in Bioanalysis was held in Long Beach, California, USA, where close to 500 professionals from pharmaceutical and biopharmaceutical companies, CROs and regulatory agencies convened to discuss current topics of interest in bioanalysis. These 'hot' topics, which covered both small and large molecules, were the starting point for fruitful exchanges of knowledge, and sharing of ideas among speakers, panelists and attendees. The discussions led to specific recommendations pertinent to bioanalytical science. Such as the previous editions, this 2013 White Paper addresses important bioanalytical issues and provides practical answers to the topics presented, discussed and agreed upon by the global bioanalytical community attending the 7th Workshop on Recent Issues in Bioanalysis.


Assuntos
Descoberta de Drogas/métodos , Animais , Bioquímica/métodos , Bioquímica/normas , Biomarcadores Farmacológicos/análise , California , Técnicas de Química Analítica/métodos , Técnicas de Química Analítica/normas , Aprovação de Drogas/métodos , Descoberta de Drogas/normas , Humanos , Farmacocinética , Estudos de Validação como Assunto
11.
Bioanalysis ; 4(18): 2213-26, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23046264

RESUMO

Over 400 professionals representing pharmaceutical companies, CROs, and multiple regulatory agencies participated in the 6th Workshop on Recent Issues in Bioanalysis (WRIB). Like the previous sessions, this event was in the format of a practical, focused, highly interactive and informative workshop aiming for high-quality, improved regulatory compliance and scientific excellence. Numerous 'hot' topics in bioanalysis of both small and large molecules were shared and discussed, leading to consensus and recommendations among panelists and attendees representing the bioanalytical community. The major outcome of this year's workshop was the noticeable alignment of multiple bioanalytical guidance/guidelines from different regulatory agencies. This represents a concrete step forward in the global harmonization of bioanalytical activities. The present 2012 White Paper acts as a practical and useful reference document that provides key information and solutions on several topics and issues in the constantly evolving world of bioanalysis.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Guias como Assunto , Imunoensaio/métodos , Espectrometria de Massas/métodos , Preparações Farmacêuticas/análise , Proteínas Recombinantes/análise , Calibragem , Cromatografia Líquida de Alta Pressão/normas , Teste em Amostras de Sangue Seco/métodos , Indústria Farmacêutica , Regulamentação Governamental , Humanos , Imunoensaio/normas , Espectrometria de Massas/normas , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Estudos de Validação como Assunto
12.
COPD ; 9(4): 322-31, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22497562

RESUMO

Chronic obstructive pulmonary disease (COPD) is characterized by inflammation of lung parenchyma and pulmonary hypoxemia with a proven systemic component. Tobacco smoke is the most important risk factor and plasma membrane plays a major role in the disease pathology and progression. The properties of biological membranes are a function of their lipid composition. Any change in its composition may lead to the pathophysiology. In COPD research, erythrocytes are emerging as a new therapeutic venture, as their shape and properties change in the disease. Therefore we studied the lipid composition of the erythrocyte membranes of COPD patients. The study included 30 patients having COPD, 10 healthy smokers and 10 non-smokers. Erythrocytes were separated from peripheral blood and their membranes prepared, followed by estimation of proteins, cholesterol and phospholipids. Individual phospholipids were identified and separated by TLC and fatty acid composition determined by gas chromatography. The data were analyzed statistically and P < 0.05 was considered significant. Our results demonstrate that in very severe COPD, proteins decrease, whereas phospholipids and cholesterol contents increase significantly, which showed a consistent negative correlation with FEV1%. The fatty acid analysis showed preponderance towards saturated fatty acids mainly arachidic and behenic acid, suggesting a decrease in membrane fluidity or a closer packing of lipid rafts. We are the first to report about preponderance of saturated fatty acids in plasma membrane of erythrocytes of COPD patients which may decrease the membrane fluidity and possibly impair the functions of the plasma membrane in the disease.


Assuntos
Membrana Eritrocítica/química , Lipídeos/sangue , Doença Pulmonar Obstrutiva Crônica/sangue , Adulto , Idoso , Biomarcadores/sangue , Estudos de Casos e Controles , Colesterol/sangue , Cromatografia Gasosa , Cromatografia Líquida , Ácidos Graxos/sangue , Feminino , Volume Expiratório Forçado , Humanos , Masculino , Pessoa de Meia-Idade , Fosfolipídeos/sangue , Doença Pulmonar Obstrutiva Crônica/etiologia , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Pesquisa Qualitativa , Fatores de Risco , Índice de Gravidade de Doença , Fumar/efeitos adversos
13.
J Asthma ; 49(2): 129-33, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22277105

RESUMO

OBJECTIVES: Erythrocyte membrane proteins reflect the prototype of multifunctional proteins of various erythroid and non-erythroid cells, which demonstrate various cellular functions. The protein profile of cells changes in various diseases. Therefore, the objective of this study was to understand the changes in protein profile of erythrocyte membranes in bronchial asthma. METHODS: The study included 20 patients of bronchial asthma and 20 healthy subjects. Erythrocytes were isolated from peripheral blood, membranes were prepared followed by the determination of protein contents, and protein profile was assessed using SDS-PAGE. RESULTS: In bronchial asthma, the protein contents of erythrocyte membranes in asthmatic patients were significantly higher (p < .005) than in healthy controls. Analysis of protein profile showed absence of the proteins, namely, band 4.2 and adducin subunit-II, and appearance of protein bands of molecular weights corresponding to galectin-3, glyceraldehyde 3-phosphate dehydrogenase, ß-actin, dematin, band 4.1, and adducin (subunit-I) in asthmatic patients when compared with healthy controls. CONCLUSIONS: In asthma, there are quantitative and qualitative changes in proteins of erythrocyte membranes. The absence of band 4.2 protein may cause impairment of the erythrocyte membrane integrity, and presence of galectin-3 may lead to the activation of various inflammatory cells. The altered protein profile may possibly lead to altered response of the inflammatory cells to the asthmogenic stimuli, which may be responsible for pathophysiology and manifestation of the symptoms of bronchial asthma.


Assuntos
Asma/sangue , Proteínas Sanguíneas/análise , Membrana Eritrocítica/química , Adolescente , Adulto , Feminino , Humanos , Masculino , Microdomínios da Membrana/metabolismo , Pessoa de Meia-Idade
14.
PLoS One ; 6(9): e25094, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21966424

RESUMO

X-linked adrenoleukodystrophy (X-ALD) affects the nervous system white matter and adrenal cortex secondary to mutations in the ABCD1 gene that encode the peroxisomal membrane protein. We conducted a genomic and protein expression study of susceptibility gene with its clinical and biochemical analysis. To the best of our knowledge this is the first preliminary comprehensive study in Indian population that identified novel mutations and SNPs in a relatively large group. We screened 17 Indian indigenous X-linked adrenoleukodystrophy cases and 70 controls for mutations and SNPs in the exonic regions (including flanking regions) of ABCD1 gene by direct sequencing with ABI automated sequencer along with Western blot analysis of its endogenous protein, ALDP, levels in peripheral blood mononuclear cells. Single germ line mutation was identified in each index case in ABCD1 gene. We detected 4 novel mutations (2 missense and 2 deletion/insertion) and 3 novel single nucleotide polymorphisms. We observed a variable protein expression in different patients. These findings were further extended to biochemical and clinical observations as it occurs with great clinical expression variability. This is the first major study in this population that presents a different molecular genetic spectrum as compared to Caucasian population due to geographical distributions of ethnicity of patients. It enhances our knowledge of the causative mutations of X-ALD that grants holistic base to develop effective medicine against X-ALD.


Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Adrenoleucodistrofia/genética , Adrenoleucodistrofia/metabolismo , Polimorfismo de Nucleotídeo Único/genética , Membro 1 da Subfamília D de Transportadores de Cassetes de Ligação de ATP , Adolescente , Western Blotting , Linhagem Celular , Criança , Pré-Escolar , Feminino , Mutação em Linhagem Germinativa/genética , Humanos , Índia , Masculino , Mutação , Mutação de Sentido Incorreto , Adulto Jovem
15.
Bioanalysis ; 3(18): 2081-96, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21942519

RESUMO

The 5th Workshop on Recent Issues in Bioanalysis (WRIB) was organized by the Calibration and Validation Group as a 2-day full immersion workshop for pharmaceutical companies, CROs and regulatory agencies to discuss, review, share perspectives, provide potential solutions and agree upon a consistent approach to recent issues in the bioanalysis of both small and large molecules. High quality, better compliance to regulations and scientific excellence are the foundation of this workshop. As in the previous editions of this significant event, recommendations were made and a consensus was reached among panelists and attendees, including industry leaders and regulatory experts representing the global bioanalytical community, on many 'hot' topics in bioanalysis. This 2011 White Paper is based on the conclusions from this workshop, and aims to provide a practical reference guide on those topics.


Assuntos
Preparações Farmacêuticas/análise , Calibragem , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia Líquida de Alta Pressão/normas , Teste em Amostras de Sangue Seco/métodos , Teste em Amostras de Sangue Seco/normas , Indústria Farmacêutica , Regulamentação Governamental , Guias como Assunto , Humanos , Espectrometria de Massas/instrumentação , Espectrometria de Massas/métodos , Espectrometria de Massas/normas , Transferência de Tecnologia , Análise Serial de Tecidos/métodos , Análise Serial de Tecidos/normas , Estudos de Validação como Assunto
16.
Cardiovasc Toxicol ; 11(4): 308-15, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21800129

RESUMO

Diabetes mellitus is very often associated with dyslipidemia, increased oxidative stress and endothelial dysfunction that could develop atherosclerosis and consequently cardiovascular diseases. Medicinal plants with reputed traditional use to treat diabetes and cardiovascular diseases might provide valuable drugs. Therefore, the present study was designed to evaluate anti-atherosclerotic potential of aqueous extract of Cassia auriculata L. leaves in streptozotocin (STZ)-induced diabetic rats. The rats were rendered diabetic by STZ (45 mg/kg, ip). Diabetic rats were orally administered C. auriculata leaf extract at 400 mg/kg dose daily for 21 days. The supplementation of extract to the diabetic rats produced significant reduction in fasting blood glucose along with significant reversal in altered serum lipid profile and apolipoprotein B. Lipid peroxidation was found to be significantly suppressed in extract-fed diabetic rats. The significant reduction in serum levels of oxidized low-density lipoprotein, soluble vascular cell adhesion molecule and plasma fibrinogen with a concomitant elevation in serum nitric oxide was observed in diabetic rats following treatment with extract. Histopathological examination of heart myocardium of extract-treated diabetic rats revealed reversal of fatty change toward normal. These results suggest that C. auriculata aqueous leaf extract exhibits anti-atherosclerotic role in the diabetic state and it indicates toward the notion that extract may help to prevent the progression of cardiovascular diseases.


Assuntos
Aterosclerose/tratamento farmacológico , Cassia/química , Diabetes Mellitus Experimental/tratamento farmacológico , Hiperglicemia/tratamento farmacológico , Hipoglicemiantes/farmacologia , Extratos Vegetais/farmacologia , Animais , Aterosclerose/etiologia , Glicemia/efeitos dos fármacos , Diabetes Mellitus Experimental/complicações , Fibrinogênio/metabolismo , Coração/efeitos dos fármacos , Hiperglicemia/complicações , Peroxidação de Lipídeos/efeitos dos fármacos , Lipoproteínas LDL/sangue , Masculino , Miocárdio/patologia , Óxido Nítrico/metabolismo , Folhas de Planta/química , Ratos , Ratos Wistar , Molécula 1 de Adesão de Célula Vascular/sangue
17.
Bioanalysis ; 3(8): 823-7, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21510754

RESUMO

Bioanalytical methods employed for the quantitative determination of drugs and their metabolites in biological fluids provide essential regulatory data for bioavailability, bioequivalence, pharmacokinetic and toxicokinetic studies. The quality of these studies is directly related to the underlying bioanalytical data. Data generated by a typical bioanalytical laboratory is submitted to not only the local regulatory agency, but also to multiple regulatory agencies worldwide. Many pharmaceutical companies and CROs are now performing bioanalytical work for global submissions and the regulatory agencies are often reviewing the bioanalytical work performed in other countries. The bioanalytical workplace has become global and therefore needs universal rules for quality and compliance of bioanalysis. This paper provides a historical perspective and insight into the development and evolution of the regulatory guidance for bioanalytical method validation and analysis of samples.


Assuntos
Testes de Química Clínica/métodos , Regulamentação Governamental/história , Preparações Farmacêuticas/análise , Animais , Testes de Química Clínica/história , Testes de Química Clínica/normas , Conferências de Consenso como Assunto , Guias como Assunto , História do Século XX , Humanos , Laboratórios , Preparações Farmacêuticas/metabolismo , Tecnologia
18.
Biochimie ; 93(7): 1146-56, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21527308

RESUMO

Investigations on the role of intracellular Ca(2+) ion concentration in the mechanism of development of COPD in smokers and non-smokers were carried out. The intracellular Ca(2+) levels were found to be increased in human lymphocytes in patients with COPD as compared to non-smokers and smokers without COPD. The investigations reveal an association in altered intracellular Ca(2+) regulation in lymphocytes and severity of COPD, by means of significant activation of Protein kinase C and inducible nitric oxide synthase (iNOS). The effect of a novel calcium channel blocker ethyl 4-(4'-heptanoyloxyphenyl)-6-methyl-3,4-dihydropyrimidin-2-one-5-carboxylate (H-DHPM) as a potential candidate for the treatment of COPD was also investigated. H-DHPM treated cells showed a decrease in intracellular Ca(2+) level as compared to the control cells. Molecular studies were carried out to evaluate the expression profile of NOS isoforms in human lymphocytes and it was shown that H-DHPM decreases the increased iNOS in COPD along with reestablishing the normal levels of endothelial nitric oxide synthase (eNOS). The results of H-DHPM were comparable with those of Amlodipine, a known calcium channel blocker. Calcium channel blocker H-DHPM proves to be a potential candidate for the treatment of COPD and further clinical studies are required to prove its role in the treatment of pulmonary hypertension (PH).


Assuntos
Bloqueadores dos Canais de Cálcio/farmacologia , Cálcio/metabolismo , Linfócitos/efeitos dos fármacos , Pirimidinonas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Bloqueadores dos Canais de Cálcio/química , Linhagem Celular , Células Cultivadas , Quelantes/farmacologia , Ácido Egtázico/farmacologia , Feminino , Citometria de Fluxo , Humanos , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Linfócitos/metabolismo , Masculino , Microscopia de Fluorescência , Pessoa de Meia-Idade , Estrutura Molecular , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Óxido Nítrico Sintase Tipo III/genética , Óxido Nítrico Sintase Tipo III/metabolismo , Proteína Quinase C/metabolismo , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/patologia , Pirimidinonas/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fumar
19.
Bioanalysis ; 2(12): 1945-60, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21110739

RESUMO

The 4th Calibration and Validation Group Workshop on Recent Issues in Regulated Bioanalysis, a 2-day full immersion workshop, was organized by the Calibration and Validation Group. Contract research organizations, pharmaceutical companies and regulatory agencies came together to discuss several 'hot' topics concerning bioanalytical issues and regulatory challenges and to reach a consensus among panelists and attendees on many points regarding method validation of small and large molecules.


Assuntos
Biofarmácia/métodos , Técnicas de Química Analítica/métodos , Cooperação Internacional , Preparações Farmacêuticas/análise , Biofarmácia/normas , Calibragem , Técnicas de Química Analítica/normas , Humanos , Preparações Farmacêuticas/normas , Controle de Qualidade , Quebeque
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