RESUMO
ABSTRACT Bovine viral diarrhea virus can cause acute disease in livestock, leading to economic losses. We show that Prostaglandin A1 inhibits bovine viral diarrhea virus replication in Madin-Darby bovine kidney cells (94% inhibition using 5 µg/mL). Light and electron microscopy of infected cells shows that Prostaglandin A1 also prevents virus-induced vacuolization, but at higher concentrations (10 µg/mL).
Assuntos
Animais , Bovinos , Antivirais/farmacologia , Prostaglandinas A/farmacologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Vírus da Diarreia Viral Bovina/efeitos dos fármacos , Antivirais/análise , Prostaglandinas A/análise , Replicação Viral/efeitos dos fármacos , Doença das Mucosas por Vírus da Diarreia Viral Bovina/tratamento farmacológico , Linhagem Celular , Vírus da Diarreia Viral Bovina/fisiologia , Vírus da Diarreia Viral Bovina/genética , DiarreiaRESUMO
Bovine viral diarrhea virus can cause acute disease in livestock, leading to economic losses. We show that Prostaglandin A1 inhibits bovine viral diarrhea virus replication in Madin-Darby bovine kidney cells (94% inhibition using 5µg/mL). Light and electron microscopy of infected cells shows that Prostaglandin A1 also prevents virus-induced vacuolization, but at higher concentrations (10µg/mL).
Assuntos
Antivirais/farmacologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Vírus da Diarreia Viral Bovina/efeitos dos fármacos , Prostaglandinas A/farmacologia , Animais , Antivirais/análise , Doença das Mucosas por Vírus da Diarreia Viral Bovina/tratamento farmacológico , Bovinos , Linhagem Celular , Diarreia , Vírus da Diarreia Viral Bovina/genética , Vírus da Diarreia Viral Bovina/fisiologia , Prostaglandinas A/análise , Replicação Viral/efeitos dos fármacosRESUMO
Production of Polyglutamate (PGA) biopolymer by immobilized Bacilluslicheniformis strain-R was intensively investigated. Preliminary experiments were carried out to address the most suitable immobilization methodology. Entrapment of Bacillus cells in alginate-agar led optimal PGA production (36.75 g/l), with 1.32- and 2.18-fold increase in comparison with alginate- or K-carrageenan-immobilized cells, respectively. During semicontinuous cultivation of agar-alginate gel-cell mixture, production of PGA by 10 ml mixture was increased from 2nd to 3rd run whereas, increased till the 4th run using 15ml mixture. Adsorption was the most suitable immobilization technique for production of PGA and the sponge cubes was the preferred matrix recording 43.2 g/l of PGA with the highest cell adsorption. Furthermore, no PGA was detected when B. licheniformis cells were adsorbed on wood and pumice. Although luffa pulp-adsorbed cells recorded the highest PGA production (50.4 g/l), cell adsorption was the lowest. Semicontinuous cultivation of B. licheniformis cells adsorbed on sponge led to increase of PGA production till the 3rd run and reached 55.5 g/l then slightly decreased in the 4th run. The successful use of fixed-bed bioreactor for semicontinuous cultivation of B. licheniformis cells held on sponge cubes (3 runs, 96 hours/run) provides insight for the potential biotechnological production of PGA by immobilized cells.
Assuntos
Reatores Biológicos , Bacillus/enzimologia , Bacillus/isolamento & purificação , Técnicas In Vitro , Poli G/análise , Poli G/biossíntese , Prostaglandinas A/análise , Prostaglandinas A/biossíntese , Meios de Cultura , Ativação Enzimática , Métodos , Reação em Cadeia da Polimerase , MétodosRESUMO
Chronic obstructive pulmonary disease (COPD) patients have increased neutrophils and macrophages in their lungs, and inflammation of the airway is related to oxidative stress. This study assessed the levels of 8-isoprostane (an oxidative stress marker) and chemokines related to neutrophil and monocyte inflammation (growth-related oncogene alpha [GROalpha] and monocyte chemoattractant protein-1 [MCP-1]) in the airway of ex-smoking COPD patients by exhaled breath condensate (EBC) collection. Thirty-two (28 males) stable COPD patients (14 with FEV(1) 50% [Group 1], 18 with FEV(1) <50% predicted [Group 2]) and 18 non-smoking age and sex-matched controls were studied in this cross-sectional study. EBC was collected using the EcoScreen (Jaeger, Germany) during 10 min of tidal breathing with the nose clipped. Concentrations of 8-isoprostane, GROalpha and MCP-1 were measured by enzyme immunoassays. COPD patients had a higher concentration of 8-isoprostane than controls (COPD versus control, P<0.001; Group 1 versus Group 2, P=0.045). 8-isoprostane increased across the groups from normal, Group 1 to Group 2 (r=0.64, P<0.001). The median intraquartile range (IQR) levels in pg/ml for GROalpha were 45.3(44.5-46.5), 45.4(44.5-46.0), 46.0(45.6-47.3), whereas MCP-1 levels were 5.3(5.2-5.9), 6.2(5.4-6.9) and 5.7(5.5-6.4) in Group 1, Group 2 COPD and control subjects, respectively. GROalpha level was lower in COPD patients when compared to controls (P=0.01). MCP-1 level did not differ between COPD and the control group. 8-isoprostane level, but not GROalpha and MCP-1, in EBC was increased in COPD patients with poorer lung function. This suggests an increased oxidative stress in the airway in patients with more severe COPD.
Assuntos
Testes Respiratórios , Quimiocina CCL2/análise , Quimiocinas CXC/análise , Peptídeos e Proteínas de Sinalização Intercelular/análise , Estresse Oxidativo/fisiologia , Prostaglandinas A/análise , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/análise , Testes Respiratórios/métodos , Quimiocina CXCL1 , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Oncogenes , Estudos Prospectivos , Doença Pulmonar Obstrutiva Crônica/fisiopatologiaRESUMO
BACKGROUND: Reliable assessment of severity in psoriasis is essential to document treatment responses in clinical research. The reliability of current clinical outcome measures is uncertain. OBJECTIVE: To quantify the relative variation in commonly used outcome measures (the Psoriasis Area and Severity Index [PASI] and one version of the Psoriasis Global Assessment [PGA]), and a newer measure, the Lattice System Physician's Global Assessment (LS-PGA). METHODS: Physicians who were experienced (53%; 9/17) or inexperienced (47%; 8/17) in using PASI and PGA evaluated 35 patients with psoriasis in random order twice with each rating system. We assessed the variation in scoring psoriasis severity within (intrarater) and among (interrater) physicians. RESULTS: PASI, PGA, and LS-PGA were highly correlated (r > 0.8 for all comparisons) and had high overall reliability (Cronbach's alpha > 0.9 for each). PGA and LS-PGA had lower intrarater variation than PASI. LS-PGA had a 55% higher concordance coefficient between the two evaluations than did PGA. Interrater variation was lower for PGA and LS-PGA than for PASI both before and after correction for measurement error. Experience was beneficial in reducing variation in PASI scores but was not required with PGA or LS-PGA. CONCLUSION: The LS-PGA, which is standardized, does not require experience, and provides discrete word-based scores with intrinsic meaning, is a reliable measure of therapeutic effect in psoriasis, and would allow comparisons across different clinical trials.
Assuntos
Padrões de Prática Médica/estatística & dados numéricos , Psoríase/classificação , Índice de Gravidade de Doença , Adulto , Superfície Corporal , Feminino , Humanos , Masculino , Michigan , Pessoa de Meia-Idade , Prostaglandinas A/análise , Psoríase/diagnóstico , Reprodutibilidade dos TestesRESUMO
Cysteinyl leukotrienes (cys-LTs), LTB4 and 8-isoprostane are increased in the exhaled breath condensate (EBC) from asthmatic patients. The aim of this study was to investigate whether the measurement of cys-LTs, LTB4 and 8-isoprostane in EBC can reflect the level of airway inflammation assessed by induced sputum in asthmatic children sensitized to house dust mite (HDM) during natural avoidance of HDM allergens. Twelve children were evaluated at the time of admission (T0) and after 3 months of stay (T1) at the Istituto Pio XII (Misurina, Italian Dolomites 1756 m). Sputum eosinophil percentage and measurement of cys-LTs, LTB4 and 8-isoprostanes in the breath condensate at T0 and T1 were evaluated. Eosinophil percentage in induced sputum was 8.5 +/- 1.1% at T0 and 3.5 +/- 0.4% at T1 (p = 0.011). Neutrophil percentage in sputum was 1.1 +/- 0.5% at T0 and 1.5 +/- 1.0% at T1 (ns). Cys-LTs mean level was 14.24 +/- 4.53 pg/ml at T0 and 4.65 +/- 0.68 pg/ml at T1 (p = 0.0125). LTB4 level was 2.36 +/- 0.19 pg/ml at T0 and 2.41 +/- 0.23 pg/ml at T1 (ns). 8-Isoprostane level reduced from 17.47 +/- 3.18 pg/ml at T0 to 7.36 +/- 3.26 pg/ml at T1 (p = 0.003). This study show that exhaled cys-LTs and 8-isoprostane, as well as eosinophil percentage in induced sputum, are reduced after allergen avoidance in asthmatic children suggesting a potential application of EBC for the non-invasive evaluation of airway inflammation in asthma in allergic asthmatic children.
Assuntos
Asma/imunologia , Testes Respiratórios/métodos , Eicosanoides/análise , Eosinófilos/imunologia , Escarro/imunologia , Adolescente , Altitude , Asma/terapia , Criança , Cisteína/análise , Cisteína/imunologia , Eicosanoides/imunologia , Feminino , Humanos , Leucotrieno B4/análise , Leucotrieno B4/imunologia , Leucotrienos/análise , Leucotrienos/imunologia , Masculino , Projetos Piloto , Prostaglandinas A/análise , Prostaglandinas A/imunologia , Pyroglyphidae/imunologiaRESUMO
AIMS: Oxidative stress is implicated in the progression of heart failure, and matrix metalloproteinase (MMP) activity is increased in patients with congestive heart failure. We examined the role of oxidative stress on MMP activity in humans. METHODS AND RESULTS: We measured the MMP activity and the level of 8-iso-prostagandin F2alpha (8-iso-PGF2alpha), a specific and quantitative maker of oxidant stress, in the pericardial fluid (PF) in 47 consecutive patients with coronary artery disease who underwent coronary artery bypass surgery. Zymography of PF showed bands at 92-85kDa (MMP-9) and 72-65kDa (MMP-2). The MMP activity was expressed as the ratio to MMP-2 standard. MMP-2, MMP-9 and total gelatinolysis activities were positively correlated with left ventricular end-diastolic volume index (LVEDVI), and MMP-2 and total gelatinolysis activities were also positively correlated with LV end-systolic volume index. Moreover, MMP-2, MMP-9 and total gelatinolysis activities were all positively correlated with pericardial level of 8-iso-PGF2alpha. Also, LVDEVI was positively correlated with pericardial level of 8-iso-PGF2alpha. CONCLUSIONS: Oxidative stress may play an important role in the regulation of MMP activity. Augmented MMP activity may be involved in the development of ventricular remodelling in patients with coronary artery disease.
Assuntos
Doença da Artéria Coronariana/etiologia , Metaloproteinases da Matriz/metabolismo , Estresse Oxidativo/fisiologia , Remodelação Ventricular/fisiologia , Idoso , Doença da Artéria Coronariana/enzimologia , Feminino , Humanos , Masculino , Pericárdio/química , Prostaglandinas A/análiseRESUMO
Peripheral vascular disease is a common ailment of the aged and diabetic communities. As the numbers of these individuals increase, the need for therapeutic interventions will continue to grow. One of the possible therapies is the use of prostaglandins (PGE(1), prostacyclin and Iloprost) to decrease the vascular tone and increase vascular blood flow. Due to the hydrophobicity of the prostaglandins and prostaglandin analogues, various vehicles have been utilized to maintain the active pharmaceutical ingredient in a stable solution, e.g. alpha-cyclodextrin (Alprostadil, Edex) or emulsified lipid vehicles. In our laboratory, we designed a method for separating and assaying lipid-encapsulated PGE(1). Utilizing organic extraction, automated solid-phase extraction and precipitation techniques, we validated the measurement of the PGE(1) and PGA(1) content of the clinical drug formulation in the microgram per milliliter concentration range with an high performance liquid chromatography (HPLC) assay.
Assuntos
Alprostadil/análise , Antivirais/análise , Prostaglandinas A/análise , Alprostadil/administração & dosagem , Antivirais/administração & dosagem , Autoanálise , Soluções Tampão , Cromatografia Líquida de Alta Pressão , Injeções Intravenosas , Lipossomos , Prostaglandinas A/administração & dosagem , Padrões de Referência , Reprodutibilidade dos Testes , Solventes , Espectrofotometria UltravioletaRESUMO
A2/J2-Isoprostanes (IsoPs) are prostaglandin (PG) A2/J2-like compounds that are produced in vivo as dehydration products of D2/E2-IsoPs. One A2-IsoP that should be formed is 15-A2t-IsoP (8-iso-PGA2). Analogous to cyclopentenone PGs, 15-A2t-IsoP readily undergoes nucleophilic addition to various biomolecules suggesting the compound is capable of exerting potent bioactivity. However, proof that it is definitively formed in vivo is lacking. Evidence is now presented that 15-A2t-IsoP, in fact, is generated in vivo by demonstrating that an endogenous A2-IsoP with a retention time on capillary GC identical with that 15-A2t-IsoP co-chromatographs through four high resolving HPLC purification procedures with authentic radiolabeled 15-A2t-IsoP.
Assuntos
Fígado/metabolismo , Prostaglandinas A/biossíntese , Animais , Tetracloreto de Carbono/toxicidade , Cromatografia Líquida de Alta Pressão , Cromatografia Gasosa-Espectrometria de Massas , Fígado/efeitos dos fármacos , Estresse Oxidativo , Prostaglandinas A/análise , Prostaglandinas A/isolamento & purificação , Ratos , Ratos Sprague-Dawley , TrítioRESUMO
A method for the simultaneous determination of prostaglandins E1, A1 and B1 (PGE1, PGA1 and PGB1) in solution has been developed by reversed-phase high-performance liquid chromatography using a 3 microns C18 column. The mobile phase consisted of 35% acetonitrile in 0.002 M phosphate buffer (pH 3.5) and its flow-rate was 1.5 ml/min. Quantitative measurement was performed using a photodiode array detector system at 190, 220, and 280 nm for PGE1, PGA1 and PGB1, respectively. The method has been applied to the primary kinetic studies for reaction profile for PGE1----PGA1----PGB1 at 60 degrees C in pH 2.0, 7.2, 10.0 and 12.0 buffer solutions.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Prostaglandinas A/análise , Prostaglandinas B/análise , Prostaglandinas E/análise , Animais , Humanos , Prostaglandinas E/farmacocinéticaRESUMO
In previous reports a direct thin-layer chromatographic method (TLC) for the determination of prostaglandins (PG) E2, A2, B2 and F2 alpha and a High-Performance Liquid Chromatographic method (HPLC) for the determination of PGE2-, PGA2- and PGB2- descendants and for stability studies of PGE2-descendant in pharmaceutical preparations were described In this paper the described HPLC method and a newly developed technique based on ion-pair HPLC are given for the simultaneous determination of PGE2, PGA2 and PGB2 and for the stability studies of PGE2 in pharmaceutical preparations. The extraction of PG's from pharmaceutical preparations is performed in a fully automated, electronically controlled extraction apparatus within 3 minutes. Ion-pair reversed phase HPLC is performed on a column of LiChrosorb RP18 using either methanol-water-octan-1-sulfonic acid sodium salt (55 ml + 45 ml + 65 mg) or methanol-water-tetrabutylammoniumperchlorate (360 ml + 290 ml + 1.0 g) as eluent. HPLC separation can also be performed on a column of mu-Bondapack C18 using methanol-n-butanol-glacial acetic acid-water (350 ml + 45 ml + 5 ml + 400 ml) as solvent. The stability of PGE2 is investigated in pure substance, in ethanolic-aqueous solution, in absolute ethanolic solution and in freeze-dried ampoules. The examined preparations are stored at temperatures between 4 and 40 degrees C and are investigated periodically. The stability studies of PGE2 indicate that PGA2 and PGB2 are formed as degradation products of PGE2. The prediction of the stability studies of PGE2 indicate that this active drug is unstable in pure substance as well as in pharmaceutical preparations.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Prostaglandinas/análise , Fenômenos Químicos , Química , Cromatografia Líquida de Alta Pressão , Dinoprostona/análise , Prostaglandinas A/análise , Prostaglandinas B/análise , Soluções , TemperaturaRESUMO
Field desorption (FD) mass spectrometry was applied to the determination of prostaglandin E2 (PGE2) in urine. Studies on synthetic compounds showed that methyl esters are stable under FD mass spectrometric conditions. The ester of PGE2 gives rise to protonated molecular ions and fragments due to the elimination of water and the side chains C8H14O and C8H12O2. This behaviour enabled its determination in the nanogram per millilitre range.
Assuntos
Prostaglandinas/análise , Dinoprostona/análise , Dinoprostona/urina , Humanos , Espectrometria de Massas , Prostaglandinas/urina , Prostaglandinas A/análise , Prostaglandinas A/urinaRESUMO
A high-performance liquid chromatographic method for the determination of prostaglandin E1 (PGE1) incorporated in white petrolatum, white ointment, hydrophilic petrolatum and Plastibase was investigated. The prostaglandin was separated from the oleaginous vehicles with n-hexane-aqueous acetonitrile. Most of white petrolatum, which is a principal component in the vehicles, remained in the n-hexane layer, and the recovery of the drug from any vehicle attained 100%. The method was applied to stability studies of PGE1 in white petrolatum and macrogol ointment. In which pure PGE1 and PGE1-alpha-cyclodextrin complex (PGE1-CD) were incorporated. The drug remained intact for up to 6 months when stored at 5 degrees C. At 25 and 40 degrees C, pure PGE1 was more stable than PGE1-CD and both PGE1 species were more stable in white petrolatum than in macrogol ointment.
Assuntos
Prostaglandinas E/análise , Alprostadil , Cromatografia Líquida de Alta Pressão/métodos , Estabilidade de Medicamentos , Bases para Pomadas , Pomadas , Veículos Farmacêuticos , Prostaglandinas A/análise , Prostaglandinas B/análise , Prostaglandinas E/isolamento & purificação , Temperatura , Fatores de TempoRESUMO
A comparison has been made between the growth patterns of two spontaneously appearing mammary adenocarcinomas in murine bone marrow radiation chimeras and in mice preimmunized with monoclonal antibodies (MAb) detecting embryo-associated antigenic determinants. A correlation was seen between the ability of the embryo-immunized chimeras to produce cytotoxic antibody to the tumors, as assessed by an antibody-dependent cellular cytotoxic assay, and the permissiveness of the mice for growth of a tumor transplant. In addition, mice deliberately preimmunized with cytotoxic MAb (antibody-dependent cellular cytotoxic assay) allowed more rapid growth specifically of that tumor earlier found to be most sensitive to the MAb used for immunization. By comparing the changing antigenic phenotype of tumor cells serially passaged through different immunized, nonimmunized mice, evidence was found suggesting that immunization could cause either antigen modulation of transferred tumor cells or a (transient) selective advantage to antigenically discrete subpopulations within the heterogeneous tumor population. Finally, we have studied the growth pattern of tumor cells transplanted into mice immunized with rabbit antibodies directed against the murine MAb. In this case, tumor growth was slowed preferentially for the tumor reactive with the specific MAb, and again, predictable changes in the antigenic spectrum of tumor cells harvested from these animals were observed. Our overall findings are interpreted in terms of the involvement of networks of antibodies reacting with embryo-associated antigens in the regulation of growth of the murine mammary adenocarcinomas studied.
Assuntos
Adenocarcinoma/imunologia , Anticorpos Monoclonais , Citotoxicidade Celular Dependente de Anticorpos , Neoplasias Mamárias Experimentais/imunologia , Prostaglandinas A/análise , Adenocarcinoma/fisiopatologia , Animais , Medula Óssea/efeitos da radiação , Quimera , Cinética , Neoplasias Mamárias Experimentais/fisiopatologia , Camundongos , Camundongos Endogâmicos , Fenótipo , Prostaglandinas A/genéticaRESUMO
An absorption high-performance liquid chromatography assay for the quantitative determination of prostaglandins A1 and B1 in alprostadil (prostaglandin E1) and in Prostin VR Pediatric Sterile Solution was developed. Prostaglandins A1 and B1 have been shown to be the major degradation products of prostaglandin E1. The adsorption system provided baseline resolution of the 2-naphthacyl esters of prostaglandin A1 from prostaglandin B1. Derivatization conditions providing maximal response for prostaglandins A1 and B1 while minimizing the conversion of prostaglandin E1 to prostaglandins A1 and B1 were established.
Assuntos
Prostaglandinas A/análise , Prostaglandinas B/análise , Prostaglandinas E/análise , Prostaglandinas/análise , Alprostadil , Cromatografia Líquida de Alta Pressão/métodos , Estabilidade de MedicamentosRESUMO
Equivalents of primary prostaglandins A and B as well as E and F were determined in the amniotic fluid of 64 clinically intact women in early pregnancy. The prostaglandin levels of the A and B groups were continuously lowered, between the ninth and 32nd weeks of pregnancy, while prostaglandin E revealed gradual rise from 57 pg/ml, in the ninth and tenth weeks, to 100 pg/ml, between the 29th and 32nd weeks. Prostaglandin F levels behaved similarly, between the 16th and 21st as well as between the 29th and 32nd weeks, mean values being 60 pg/ml or 75 pg/ml, respectively. High prostaglandin F levels, as recorded between the ninth and twelfth weeks of pregnancy (244 pg/ml), were probably attributable to mode by which amniotic fluid has been collected. Collection in that phase of pregnancy was transcervically, following cervix dilation during abortion, which differed from transabdominal amniocentesis in more advanced pregnancy. The above findings are discussed with regard to their physiological and clinical relevance.