RESUMO
OBJECTIVE: To assess prospectively whether preimplantation B-type natriuretic peptide (BNP) and C reactive protein (CRP) concentrations predict future appropriate therapies from an implantable cardioverter-defibrillator (ICD). DESIGN AND SETTING: Prospective cohort study conducted in a tertiary cardiac care centre. METHODS: 345 consecutive patients undergoing first time ICD implantation were prospectively studied. Serum BNP and CRP concentrations were obtained the day before ICD implantation. Patients were followed up with device interrogation to assess for appropriate shocks or antitachycardia pacing. Inappropriate therapies were excluded. Mean (SD) follow up was 13 (5) months. RESULTS: Patients had ischaemic (71%), primary dilated (17%), and valvar or other cardiomyopathies (12%). About half (52%) had ICDs implanted for primary prevention. Sixty three (18%) received appropriate ICD therapies. Serum creatinine, beta blocker, statin, and angiotensin converting enzyme inhibitor usage did not differ between therapy and no therapy groups. By univariate comparison, ejection fraction (p = 0.048), not taking amiodarone (p = 0.033), and BNP concentration (p = 0.0003) were risk factors for ICD therapy. However, by Cox regression multivariate analysis, only BNP above the 50th centile was a significant predictor (hazard ratio 2.19, 95% confidence interval 1.07 to 4.71, p = 0.040). Median BNP was 573 ng/l versus 243 ng/l in therapy and no therapy patients, respectively (p = 0.0003). More patients with BNP above the 50th centile (27% v 10%, p = 0.006) received ICD therapies. CONCLUSIONS: A single preimplantation BNP concentration determination is independently predictive of ICD therapies in patients with cardiomyopathies undergoing first time ICD implantation. CRP was not independently predictive of ICD therapies when compared with BNP.
Assuntos
Doença da Artéria Coronariana/terapia , Desfibriladores Implantáveis/estatística & dados numéricos , Peptídeo Natriurético Encefálico/sangue , Biomarcadores/sangue , Proteína C-Reativa/análise , Métodos Epidemiológicos , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeAssuntos
Giardíase/complicações , Hemorragia/etiologia , Síndromes de Malabsorção/etiologia , Vitamina K/metabolismo , Animais , Anticoagulantes/efeitos adversos , Diagnóstico Diferencial , Feminino , Giardia lamblia , Giardíase/diagnóstico , Giardíase/tratamento farmacológico , Humanos , Pessoa de Meia-Idade , Varfarina/efeitos adversosRESUMO
OBJECTIVES: We sought to determine the electrocardiographic (ECG) features associated with acute left main coronary artery (LMCA) obstruction. BACKGROUND: Prediction of LMCA obstruction is important with regard to selecting the appropriate treatment strategy, because acute LMCA obstruction usually causes severe hemodynamic deterioration, resulting in a less favorable prognosis. METHODS: We studied the admission 12-lead ECGs in 16 consecutive patients with acute LMCA obstruction (LMCA group), 46 patients with acute left anterior descending coronary artery (LAD) obstruction (LAD group) and 24 patients with acute right coronary artery (RCA) obstruction (RCA group). RESULTS: Lead aVR ST segment elevation (>0.05 mV) occurred with a significantly higher incidence in the LMCA group (88% [14/16]) than in the LAD (43% [20/46]) or RCA (8% [2/24]) groups. Lead aVR ST segment elevation was significantly higher in the LMCA group (0.16 +/- 0.13 mV) than in the LAD group (0.04 +/- 0.10 mV). Lead V(1) ST segment elevation was lower in the LMCA group (0.00 +/- 0.21 mV) than in the LAD group (0.14 +/- 0.11 mV). The finding of lead aVR ST segment elevation greater than or equal to lead V(1) ST segment elevation distinguished the LMCA group from the LAD group, with 81% sensitivity, 80% specificity and 81% accuracy. A ST segment shift in lead aVR and the inferior leads distinguished the LMCA group from the RCA group. In acute LMCA obstruction, death occurred more frequently in patients with higher ST segment elevation in lead aVR than in those with less severe elevation. CONCLUSIONS: Lead aVR ST segment elevation with less ST segment elevation in lead V(1) is an important predictor of acute LMCA obstruction. In acute LMCA obstruction, lead aVR ST segment elevation also contributes to predicting a patient's clinical outcome.
Assuntos
Estenose Coronária/diagnóstico , Vasos Coronários , Eletrocardiografia/métodos , Doença Aguda , Adulto , Idoso , Análise de Variância , Circulação Colateral , Angiografia Coronária , Estenose Coronária/classificação , Estenose Coronária/mortalidade , Estenose Coronária/fisiopatologia , Estenose Coronária/terapia , Análise Discriminante , Eletrocardiografia/instrumentação , Eletrocardiografia/normas , Feminino , Hemodinâmica , Humanos , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Seleção de Pacientes , Valor Preditivo dos Testes , Prognóstico , Estudos Retrospectivos , Sensibilidade e Especificidade , Resultado do TratamentoRESUMO
We have previously shown [Sakai et al., J. Biosci. Bioeng., 88, 306-309 (1999)] that exogenously supplied phosphatidic acid (PA) and lysophosphatidic acid (LPA) promoted the growth of Chinese hamster ovary (CHO) cells in serum-free culture. However, the direct addition of high concentrations of these phospholipids alone to the culture medium resulted in the formation of precipitates. We therefore examined the use of two nonionic surfactants, Tween 80 and Pluronic F-68, as a means of supplying PA more effectively to CHO cells in a serum-free culture. A clear dispersion of PA from egg yolk lecithin that could be successfully sterile-filtered was obtained by using Tween 80 or Pluronic F-68. When PA prepared with either of the surfactants was added to serum-free media, precipitation was noticeably reduced. Furthermore, the growth-promoting activity of PA was considerably enhanced by the presence of the surfactants. Since Tween 80 and Pluronic F-68 themselves possessed no growth-stimulating property, it was suggested that the enhanced growth-promoting activity results from the improved availability of PA to the cells. The use of Tween 80 with PA analogues having saturated acyl chains also accelerated cell growth, whereas these PAs showed little growth-promoting activity, due to their poor water-solubility, when added alone.
RESUMO
Severe mitral regurgitation was associated with cardiogenic shock in five (0.8%) of 623 patients with acute myocardial infarction who were urgently admitted to the authors' hospitals between 1994 and 1996. The infarct was located in the inferior wall in four patients and in the inferoposterior wall in one patient. Severe mitral valve regurgitation occurred concurrently with cardiogenic shock between one and six days after the onset of myocardial infarction. A mitral regurgitant murmur was not audible in four of five patients. Similarly, mitral regurgitant Doppler signals were not detected in four patients by transthoracic echocardiographic examination, while transesophageal echocardiographic examination detected mitral regurgitant signals clearly in all patients. Thus, when cardiogenic shock is unexpectedly associated with inferior or inferoposterior wall acute myocardial infarction, severe mitral regurgitation should be suspected, even when a mitral regurgitant murmur is not audible. Furthermore, mitral regurgitant flow signals may not always be detected by transthoracic echocardiography. Thus, examination for mitral regurgitation by transesophageal echocardiography should be considered.
Assuntos
Ecocardiografia Transesofagiana , Ruptura Cardíaca Pós-Infarto/diagnóstico por imagem , Insuficiência da Valva Mitral/diagnóstico por imagem , Infarto do Miocárdio/diagnóstico por imagem , Músculos Papilares/diagnóstico por imagem , Idoso , Diagnóstico Diferencial , Ecocardiografia Doppler , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Choque Cardiogênico/diagnóstico por imagemRESUMO
Immobilization of insect cells using porous biomass support particles (BSPs) and production of a recombinant protein by the immobilized cells after infection with a baculovirus were investigated in a shake-flask culture. Sf9 cells were passively immobilized in reticulated polyvinyl formal (PVF) resin BSPs (2 x 2 x 2 mm cubes) with matrices of 60 mum mean pore diameter in situ in shake-flasks. The cell density in the BSPs was over 5 x 10(7) cells/cm3-BSP in cultures with regular replacement of the culture medium, as estimated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. After infection with a recombinant baculovirus carrying the beta-galactosidase gene, immobilized cells within the BSPs showed a high specific productivity, comparable to the maximum productivity in shake-flask cultures of non-immobilized cells, as long as nutrients in the medium were not depleted. Even when immobilized cells at a high density of 5 x 10(7) cells/cm3-BSP were infected with the baculovirus, efficient beta-galactosidase production with a high specific productivity was possible by replacing the medium at appropriate intervals to avoid nutrient depletion.
RESUMO
The interactions of four HIV-protease inhibitors, ritonavir (RIT), saquinavir (SAQ), indinavir (IND) and nelfinavir (NEL), were examined by in vitro metabolic studies using rat liver microsomal fractions. The substrate concentrations employed were 0.75 approximately 12 microM, and the inhibitor concentrations were 2.5 approximately 60 microM. The metabolic clearance rates of SAQ, NEL and IND as determined by V(max)/K(m) were 170.9+/-10.9, 126.0+/-4.4 and 73.0+/-2.0 microL/min/mg protein, respectively. RIT was a potent inhibitor of the other three protease inhibitors, and the inhibition constants (K(i)) were 1.64 microM for SAQ, 0.95 microM for IND and 1. 01 microM for NEL. NEL was the second strongest inhibitor with a K(i) for NEL inhibition of IND metabolism of 2.14 microM. IND was the third strongest inhibitor with K(i)s of 2.76 microM for inhibition of NEL and 3.55 microM for inhibition of SAQ. As SAQ has the highest metabolic clearance rate, the K(i) for the SAQ inhibition of IND metabolism was high, 9.50 microM. Based on these in vitro results, drug interactions between NEL and IND or RIT were studied after oral administration to rats where the dose of each drug was 20 mg/kg. The C(max) and AUC of NEL were increased 3.6- and 8.5-fold by the co-administration with RIT. However, in contrast to co-administration of NEL and RIT, the effect of IND on the pharmacokinetics of NEL was negligible and the t(1/2) of NEL was not significantly increased by IND. Therefore, the combination of NEL and IND is recommended as a combination therapy for AIDS patients.
Assuntos
Inibidores da Protease de HIV/farmacocinética , Síndrome da Imunodeficiência Adquirida/tratamento farmacológico , Animais , Interações Medicamentosas , Quimioterapia Combinada , Indinavir/farmacocinética , Masculino , Nelfinavir/farmacocinética , Ratos , Ratos Wistar , Ritonavir/farmacocinética , Saquinavir/farmacocinéticaRESUMO
Mutations in the transforming growth factor-beta type II receptor (RII) gene that remain uncorrected due to mutation and inactivation of mismatch repair genes play an important role in hereditary nonpolyposis colorectal cancer (HNPCC) and in a subset of sporadic colorectal cancers. Some colorectal cancers develop from colorectal polyps. To elucidate the role of the RII gene in the generation of colorectal polyps, we analyzed 137 colorectal polyps from 100 patients for RII mutations and microsatellite instability (MSI). MSI was detected in three of 36 polyps from 25 patients. For one of these three polyps, the mobilities of the PCR products between polyp and nonpolyp tissues was different for only one microsatellite marker, and for the other two polyps the mobilities were different for more than two markers. These two polyps were obtained from one patient with ascending colon carcinoma and suspected HNPCC based on his clinical profile and family history. An RII mutation was detected in only one of these two polyps. RII may play a minor role in sporadic colorectal polyps. RII gene analysis in colorectal polyps may be a useful screening measure for potential HNPCC patients.
Assuntos
Pólipos do Colo/genética , Pólipos Intestinais/genética , Mutação/fisiologia , Receptores de Fatores de Crescimento Transformadores beta/genética , Neoplasias Retais/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Pólipos do Colo/patologia , Feminino , Humanos , Pólipos Intestinais/patologia , Masculino , Repetições de Microssatélites , Pessoa de Meia-Idade , Linhagem , Proteínas Serina-Treonina Quinases , Receptor do Fator de Crescimento Transformador beta Tipo II , Neoplasias Retais/patologiaRESUMO
The effects of dose on the pharmacokinetic characteristics of KNI-272 were evaluated in rats after intravenous (iv) administration. The plasma kinetics of KNI-272 were dose-independent within a dose range of 1.0 to 10.0 mg/kg. However, when the dose was increased to 50.0 mg/kg, the area under the plasma concentration-time curve (AUC)/dose significantly increased and the total plasma clearance (Cl(tot)) significantly decreased, possibly due to saturation of hepatic metabolism. On the other hand, the terminal elimination half-life (t(1/2,lambda(z))) was independent of dose. Using biochemical and physiological parameters obtained from in vitro and in vivo studies, we developed a physiologically based pharmacokinetic (PBPK) model for KNI-272 in rats in which concentration-dependent nonlinear hepatic metabolism (Michaelis-Menten type metabolism) was considered. Using this PBPK model, plasma KNI-272 concentration-time profiles were simulated. From these profiles it was demonstrated that the terminal elimination phase was proportional to the dose at lower doses. However, as the dose increased to 50.0 mg/kg, the simulated plasma concentrations at the terminal elimination phase increased more than the increase of dose in the same way as the observed data. Accordingly, the dose-dependent plasma kinetics observed after a 50.0 mg/kg dose was considered to be attributable in part to concentration-dependent hepatic metabolism in rats.
Assuntos
Fármacos Anti-HIV/farmacocinética , Inibidores da Protease de HIV/farmacocinética , Oligopeptídeos/farmacocinética , Animais , Fármacos Anti-HIV/sangue , Relação Dose-Resposta a Droga , Inibidores da Protease de HIV/sangue , Injeções Intravenosas , Masculino , Microssomos Hepáticos/metabolismo , Modelos Biológicos , Oligopeptídeos/sangue , Valor Preditivo dos Testes , Ratos , Ratos Wistar , Distribuição TecidualRESUMO
Upper respiratory tract infection including chronic tonsillitis is considered to be involved in the onset and/or the progression of IgA nephropathy. It is well known that deterioration of urinary findings occurs after episodes of upper respiratory tract infection in patients with IgA nephropathy. We previously showed that the expression of macrophage-colony-stimulating factor (M-CSF) is increased in the glomeruli of patients with IgA nephropathy and correlated with glomerular mesangial proliferation, suggesting that M-CSF plays an important role in the progression of IgA nephropathy. In the present study, we measured the serum and urinary concentrations of M-CSF in patients with IgA nephropathy associated with chronic tonsillitis. Furthermore, we evaluated the effects of the local provocation test of tonsils (mechanical tonsil stimulation) on the serum and urinary concentrations of M-CSF in the following three groups: (1) IgA nephropathy with severe mesangial proliferation, (2) IgA nephropathy with mild mesangial proliferation, and (3) patients with chronic tonsillitis without renal disease. The serum and urinary levels of M-CSF in the groups with severe and mild IgA nephropathy were significantly higher than those in the chronic tonsillitis group. The urinary M-CSF level but not the serum M-CSF level was positively correlated with the degrees of mesangial proliferation and glomerular M-CSF expression in the renal biopsy specimens. The urinary M-CSF concentration was significantly increased after tonsillitis stimulation in both mild and severe IgA nephropathy groups. Enhanced urinary excretion of M-CSF prolonged for 7 days after tonsil stimulation in the severe IgA nephropathy group; in contrast, the urinay M-CSF level was increased for only 2 days after tonsil stimulation in the mild IgA nephropathy group. The urinary M-CSF level was not changed in the chronic tonsillitis group after tonsil stimulation. The serum concentrations of M-CSF were not changed after tonsil stimulation in these three groups. Our present results suggest that tonsil stimulation contributes to the progression of IgA nephropathy via enhancement of glomerular production of M-CSF. The urinary excretion of M-CSF may be a useful predictor to evaluate the relevance of chronic tonsillitis to the disease and the indication of tonsillectomy in patients with IgA nephropathy.
Assuntos
Glomerulonefrite por IGA/complicações , Glomerulonefrite por IGA/urina , Fator Estimulador de Colônias de Macrófagos/urina , Tonsila Palatina/fisiopatologia , Tonsilite/complicações , Adulto , Estudos de Casos e Controles , Feminino , Mesângio Glomerular/metabolismo , Glomerulonefrite por IGA/fisiopatologia , Humanos , Fator Estimulador de Colônias de Macrófagos/sangue , Fator Estimulador de Colônias de Macrófagos/metabolismo , Masculino , Pessoa de Meia-Idade , Estimulação Física , Tonsilite/fisiopatologiaRESUMO
KNI-272 is a tripeptide protease inhibitor for treating human immunodeficiency virus type 1 (HIV-1). In in vitro stability studies using rat tissue homogenates, KNI-272 concentrations in the liver, kidney, and brain decreased significantly with time. Moreover, in tissue distribution studies, KNI-272 distributed highly to the liver, kidney, and small intestine in vivo. From these results and reported physiological parameters such as the tissue volume and tissue blood flow rate, we considered the liver to be the main organ which takes part in the metabolic elimination of KNI-272. Then the hepatic metabolism of KNI-272 was more thoroughly investigated by using rat liver microsomes. KNI-272 was metabolized in the rat liver microsomes, and five metabolites were found. The initial metabolic rate constant (kmetabolism) tended to decrease when the KNI-272 concentration in microsomal suspensions increased. The calculated Michaelis-Menten constant (K(m)) and the maximum velocity of KNI-272 metabolism (Vmax), after correction for the unbound drug concentration, were 1.12 +/- 0.09 micrograms/ml (1.68 +/- 0.13 microM) and 0.372 +/- 0.008 microgram/mg of protein/min (0.558 +/- 0.012 nmol/mg of protein per min), respectively. The metabolic clearance (CLint,metabo), calculated as Vmax/K(m), was 0.332 ml/mg of protein per min. Moreover, by using selective cytochrome P-450 inhibitors and recombinant human CYP3A4 fractions, KNI-272 was determined to be metabolized mainly by the CYP3A isoform. In addition, ketoconazole, a representative CYP3A inhibitor, inhibited KNI-272 metabolism competitively, and the inhibition constant (Ki) was 4.32 microM.
Assuntos
Inibidores da Protease de HIV/metabolismo , HIV-1 , Microssomos Hepáticos/metabolismo , Oligopeptídeos/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Sistema Enzimático do Citocromo P-450/química , Sistema Enzimático do Citocromo P-450/metabolismo , Inibidores da Protease de HIV/farmacocinética , Humanos , Técnicas In Vitro , Injeções Intravenosas , Isoenzimas/química , Isoenzimas/metabolismo , Masculino , Oligopeptídeos/farmacocinética , Ratos , Ratos Wistar , Distribuição TecidualRESUMO
The development of fluorescence bronchoscopy has made it possible to diagnose locally superficial hilar lung cancers such as carcinoma in situ, and in some cases atypical squamous metaplasia thought to be precancerous lesions. The selection of a treatment modality can be difficult for early hilar lung cancers due to problems associated with multiple lung carcinomas and the large number of heavy smokers afflicted. Many patients also have decreased pulmonary function. If carcinoma in situ and microinvasive carcinomas can be detected early, it may be possible to treat them radically with a less invasive method than surgery, such as endoscopic laser therapy. Centrally arising squamous cell carcinoma of the tracheobronchial tree, especially in heavy smokers, is thought to develop in multiple stages from squamous metaplasia, to atypical squamous metaplasia, followed by carcinoma in situ, and finally invasive cancer. However, it is hoped that preventive medicine for lung cancer will be established whereby patients with localized atypical squamous metaplasia detected by fluorescence bronchoscopy can be carefully monitored and motivated to stop smoking, and also administered chemopreventive agents.
Assuntos
Broncoscopia , Neoplasias Pulmonares/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma in Situ/patologia , Carcinoma de Células Escamosas/patologia , Feminino , Fluorescência , Humanos , Masculino , Metaplasia , Pessoa de Meia-Idade , Lesões Pré-Cancerosas/patologiaRESUMO
Sf9 insect cells infected with a recombinant baculovirus expressing beta-galactosidase and suspended in fresh medium (TNM-FH supplemented with 10% fetal bovine serum) at the time of infection were cultured in shake flasks at various combinations of initial cell density and multiplicity of infection (MOI). The effects of cell density and MOI on beta-galactosidase production were quantitatively analyzed by plotting the beta-galactosidase yield against the time integral of the viable cell density from the time of infection to the time when the beta-galactosidase production reached a plateau. The beta-galactosidase yield had a maximum value at a viable cell density time integral of approximately 8 x 10(6) cells.d/cm3 for each MOI used in a range from 0.01 to 10 plaque-forming units per cell (pfu/cell). Since glucose and fructose were exhausted when the culture reached 8 x 10(6) cells.d/cm3, it was concluded that protein production in a high-cell-density culture was limited by nutrient depletion in the culture medium, and hence the nutritional capacity of the medium was able to be determined as the viable cell density time integral at which the maximum product yield was attained. In cultures infected at a low MOI (< or =1 pfu/cell), the specific productivity, and thereby the yield, of beta-galactosidase declined with decreasing MOI due to the reduction in the proportion of initially infected cells. These results indicate that production of a recombinant protein in a culture with medium replacement at the time of infection can be optimized if the cells are infected at a high MOI (> or = 1 pfu/cell) and at a cell density such that the viable cell density time integral reaches the nutritional capacity just as the protein production is completed.
RESUMO
The effects of phosphatidic acid (PA) and lysophosphatidic acid (LPA) dispersed in serum-free media on the growth of Chinese hamster ovary (CHO) cells were investigated. After cells were incubated in serum-free media supplemented with PA or LPA for 3 d, the cellular growth was evaluated by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Supplementing PA from egg yolk lecithin to basal synthetic media such as alpha-MEM (minimum essential medium, alpha modification) markedly promoted the growth of CHO cells. PA from egg yolk lecithin also enhanced the cell growth in a serum-free medium containing insulin and transferring. When PA analogues with different acyl chains of C14-18 were compared, PA with unsaturated acyl chains, dioleoyl-PA (C18: 1), was most effective for stimulating the growth of CHO cells. Similar results were obtained when LPA was examined. These results suggest that PA and LPA, especially those with unsaturated acyl chains, are promising growth-promoting supplements for use as constituents in a low-protein serum-free medium.
Assuntos
Glomerulonefrite Membranosa/complicações , Glomerulonefrite Membranosa/patologia , Glomérulos Renais/patologia , Psoríase/complicações , Albuminúria , Membrana Basal/patologia , Membrana Basal/ultraestrutura , Glomerulonefrite Membranosa/tratamento farmacológico , Humanos , Glomérulos Renais/ultraestrutura , Masculino , Pessoa de Meia-Idade , Prednisolona/uso terapêutico , Proteinúria , Psoríase/tratamento farmacológico , Psoríase/patologiaRESUMO
A 58-year-old man with idiopathic dilated cardiomyopathy was treated with incremental administration of a beta-blocker (metoprolol) and an angiotensin converting enzyme inhibitor (enarapril). The left ventricular end-diastolic dimension and ejection fraction improved in 8 months from 83.3 mm and 17.3% to 46 mm and 69%, respectively. The washout ratio and heart-to-mediastinum ratio depicted on the delayed image for iodine-123 metaiodobenzylguanidine 123I-MIBG) uptake improved from 61.7% and 1.34 to 23.1% and 1.85, respectively, in association with improvement of left ventricular indices. Successive endomyocardial biopsy specimens disclosed reduction of the degrees of vacuolation, staining irregularity, and deformity of myocyte cytoplasm and nucleus compared to the findings before therapy. In this patient with dilated cardiomyopathy 123I-MIBG scintigraphy was useful for the evaluation of the effects of therapy. We conclude that it may be informative in the estimation of the histopathological abnormalities of the myocardium.
Assuntos
Antagonistas Adrenérgicos beta/administração & dosagem , Inibidores da Enzima Conversora de Angiotensina/administração & dosagem , Cardiomiopatia Dilatada/diagnóstico por imagem , Cardiomiopatia Dilatada/tratamento farmacológico , Enalapril/administração & dosagem , Radioisótopos do Iodo , Iodobenzenos , Metoprolol/administração & dosagem , Função Ventricular Esquerda , 3-Iodobenzilguanidina , Biópsia , Cardiomiopatia Dilatada/fisiopatologia , Endocárdio/patologia , Humanos , Radioisótopos do Iodo/farmacocinética , Iodobenzenos/farmacocinética , Masculino , Pessoa de Meia-Idade , Miocárdio/patologia , Cintilografia , Volume SistólicoRESUMO
The growth kinetics of anchorage-independent animal cells [mouse myeloma MPC-11 (ATCC CCL 167)] immobilized within porous polyvinyl formal resin biomass support particles (BSPs; 3 x 3 x 3 mm cubes; mean pore diameter, 60 microns) was analyzed by measuring intracellular and extracellular lactate dehydrogenase (LDH) activities in a perfusion culture. First, the intracellular LDH content of cells immobilized within the BSPs was assayed in a shake-flask culture and found to remain almost comparable to that of non-immobilized cells in the exponential growth phase under static culture. Then, cells inoculated in the BSPs were grown in a continuous stirred-tank bioreactor. Using the LDH content of non-immobilized cells, the density of immobilized cells and the numbers of leaked and dead cells were evaluated, respectively, by the intracellular LDH activity of immobilized and leaked cells and the LDH activity in cell-free culture supernatant. In the initial period, immobilized cells exhibited exponential growth at a constant apparent specific growth rate; however, the actual specific growth rate, which takes into consideration cell death and cell leakage, decreased significantly. In the stationary phase, the actual specific growth rate maintained a constant but markedly lower value than during exponential growth.
Assuntos
Divisão Celular , Animais , Biotecnologia , Adesão Celular , Contagem de Células , Morte Celular , Meios de Cultura , Técnicas Citológicas , Cinética , L-Lactato Desidrogenase/metabolismo , Camundongos , Células Tumorais Cultivadas/enzimologia , Células Tumorais Cultivadas/patologiaRESUMO
OBJECTIVE: We describe a monochromatic X-ray CT system using synchrotron radiation, which has been constructed employing a vertical wiggler beam line at the National Laboratory for High-Energy Physics in Tsukuba, Japan. This CT system was produced to facilitate examination of the microstructures of material and biological specimens. MATERIALS AND METHODS: The system provides excellent contrast and has a spatial resolution capacity ranging from 2 to 100 mm. In this preliminary experiment, the images of the rat were obtained at 36-mm slice thickness and 33.2-keV X-ray energy. RESULTS: The nasal cartilage, skull, and teeth of a live rat were clearly demonstrated with 36-mm spatial resolution. CONCLUSION: Using a monochromatic X-ray CT system with synchrotron radiation, detailed structures of rat skull were imaged clearly.
Assuntos
Síncrotrons , Tomografia Computadorizada por Raios X/métodos , Animais , Cartilagem/diagnóstico por imagem , Nariz/diagnóstico por imagem , Ratos , Dente/diagnóstico por imagemRESUMO
Growth and death of anchorage-independent animal cells entrapped within porous biomass support particles (BSPs) in static or shake-flask cultures were evaluated by comparison of enzyme activity with non-immobilized cells grown under static culture using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and release of lactate dehydrogenase into the culture medium. Mouse myeloma MPC-11 (ATCC CCL 167) cells inoculated within porous polyvinyl formal resin BSPs (3 x 3 x 3 or 2 x 2 x 2 mm; mean pore diameter, 60 microns) grew exponentially at a specific growth rate comparable to that of non-immobilized cells in the initial period of incubation. Entrapped cells then reached the stationary phase with a cell density over 10(7) cells/cm3 BSP. The death rate of entrapped cells increased in response to the rise in viable cell density in the BSPs. Observation of viable cell distribution within the BSPs using MTT staining indicated that the cells concentrated within a thin outer shell of the BSPs with time. After the immobilized cells reached the stationary phase, penetration of cells into the outer shell ceased and heterogeneous distribution of cell density occurred in the viable cell layer in the shake-flask culture.
Assuntos
Morte Celular , Divisão Celular , Células Tumorais Cultivadas/citologia , Animais , Contagem de Células , Corantes , L-Lactato Desidrogenase/análise , Camundongos , Polivinil , Resinas Vegetais , Sais de Tetrazólio , TiazóisRESUMO
The use of nondwarf rice (Oryza sativa L.) cultivars treated with uniconazole as test plants for gibberellin (GA) bioassay instead of Tan-ginbozu dwarf rice variant was investigated. The sensitivity of six nondwarf rice cultivars to GAs was increased substantially by treatment of the seeds with uniconazole. The minimum detectable dose of a GA in the nondwarf cultivars treated with uniconazole was 1- to 1/10-fold of that in the nontreated Tanginbozu and 3- to 10-fold of that in uniconazole-treated Tanginbozu. The relative activity of several GAs on treated nondwarf rice cultivars was not largely different from that to Tan-ginbozu. Considering that seeds of nondwarf rice are available commercially, the nondwarf rice seedling assay would be useful as a simple assay for systematic analysis of GAs, and also as a routine teaching tool in high schools and universities.