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1.
Int J Obstet Anesth ; 49: 103235, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34810053

RESUMO

BACKGROUND: The obstetric quality of recovery scoring tool (ObsQoR-11) was developed and validated in the United Kingdom for use after elective and emergency caesarean delivery. Quality of recovery scoring tools validated in one country may not be valid in other countries with significant cultural, socio-economic and linguistic variations. The aim of the current study was to validate a Hindi version of the ObsQoR-11. METHODS: In this prospective observational study, 100 parturients who underwent elective caesarean delivery in a tertiary care obstetric referral university hospital in North India were asked to complete a Hindi version of the ObsQoR-11 scoring tool 24 h after surgery. The performance of the Hindi version of ObsQoR-11 was assessed using measures of validity, reliability, and feasibility. RESULTS: The Hindi version of ObsQoR-11 correlated moderately with the global health visual analogue scale (r=0.45, 95% CI 0.27 to 0.59; P <0.0001) and discriminated well between good and poor recovery (mean (SD) score 84.6 (9.4) vs 75.0 (11.2); P <0.0001). The reliability and internal consistency were moderate (Cronbach's alpha=0.66; Spearman-Brown Prophesy Reliability estimate=0.57) with good repeatability (intraclass correlation coefficient 0.85, 95% CI 0.69 to 0.93; P <0.0001) and no floor or ceiling effects. All parturients completed the questionnaire in a (median (IQR) time of completion of 3 (1.5 - 5.5) min). CONCLUSION: The Hindi version of the ObsQoR-11 questionnaire is a promising scoring tool to evaluate quality of recovery after elective caesarean delivery. Further research is needed to evaluate the Hindi tool in other institutions in India as well as in other languages.


Assuntos
Cesárea , Países em Desenvolvimento , Feminino , Humanos , Gravidez , Estudos Prospectivos , Psicometria , Reprodutibilidade dos Testes , Inquéritos e Questionários
2.
Genomics ; 112(1): 108-113, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-30735793

RESUMO

The study was undertaken to decipher the microRNA (miRNA) related markers associated with corpus luteum (CL) tropism in buffalo. The data obtained from deep sequencing of CL tissue from different physiological stages was mined in silico for the identification of miRNA-related markers (SSR & SNP). From the present study, 5 annotated and 176 unannotated miRNA were deduced while comparing with Bos taurus genome. In addition, 4 SSRs and 9 SNPs were deduced from the miRNA sequences. These SSRs were on the genes viz. Eukaryotic translation initiation factor 1-like, myocyte enhancer factor 2A, beta casein, T cell receptor gamma cluster 1. The SNP positions on genes viz. PYGO1 (Pygopus family PHD finger 1), LOC100337244 (Multidrug resistance-associated protein 4), FTH1 (Ferritin heavy chain 1), LOC788634 (BOLA class I histocompatibility antigen), PLXND1 (Plexin D1) and UBC (Ubiquitin C) show that these genes play critical role in CL tropism during estrous cycle in buffalo.


Assuntos
Búfalos/genética , Corpo Lúteo/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Animais , Biomarcadores/metabolismo , Búfalos/metabolismo , Feminino , Regulação da Expressão Gênica , MicroRNAs/química , Repetições de Microssatélites , Polimorfismo de Nucleotídeo Único , Gravidez
3.
Int J Obstet Anesth ; 33: 90, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-29126861

Assuntos
Poaceae , Humanos
5.
Cytotechnology ; 68(6): 2271-2285, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27650183

RESUMO

The study examined the effects of different environmental stress on developmental competence and the relative abundance (RA) of various gene transcripts in oocytes and embryos of buffalo. Oocytes collected during cold period (CP) and hot period (HP) were matured, fertilized and cultured in vitro to blastocyst hatching stage. The mRNA expression patterns of genes implicated in developmental competence (OCT-4, IGF-2R and GDF-9), heat shock (HSP-70.1), oxidative stress (MnSOD), metabolism (GLUT-1), pro-apoptosis (BAX) and anti-apoptosis (BCL-2) were evaluated in immature and matured oocytes as well as in pre-implantation stage embryos. Oocytes reaching MII stage, cleavage rates, blastocyst yield and hatching rates increased (P < 0.05) during the CP. In MII oocytes and 2-cell embryos, the RA of OCT-4, IGF-2R, GDF-9, MnSOD and GLUT-1 decreased (P < 0.05) during the HP. In 4-cell embryos, the RA of OCT-4, IGF-2R and BCL-2 decreased (P < 0.05) in the HP, whereas GDF-9 increased (P < 0.05). In 8-to 16-cell embryos, the RA of OCT-4 and BCL-2 decreased (P < 0. 05) in the HP, whereas HSP-70.1 and BAX expression increased (P < 0.05). In morula and blastocyst, the RA of OCT-4, IGF-2R and MnSOD decreased (P < 0.05) during the HP, whereas HSP-70.1 was increased (P < 0.05). In conclusion, deleterious seasonal effects induced at the GV-stage carry-over to subsequent embryonic developmental stages and compromise oocyte developmental competence and quality of developed blastocysts.

6.
Anim Reprod Sci ; 152: 26-31, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25497424

RESUMO

The variety of mammalian cells has been successfully cryopreserved by use of the silk protein sericin due to its strong free-radical-scavenging and potent antioxidant activity. The present study was conducted to examine the protective role of sericin on buffalo spermatozoa during cryopreservation. Semen of four breeding bulls was collected twice a week using artificial vagina technique. The ejaculates of four bulls were pooled, divided into five equal fractions, diluted with the extender supplemented with different concentrations of sericin (0, 0.25, 0.5, 1.5 and 2%) and then cryopreserved. Post-thawed motility was objectively assessed by computer assisted sperm analyzer. Sperm plasma membrane integrity was assessed by hypo-osmotic swelling test (HOST). Malondialdehyde (MDA) concentration, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were determined in frozen-thawed extended seminal plasma by spectrophotometry. The extender supplemented with 0.25, 0.5 and 1% sericin resulted in the higher sperm motility and GPx acivity. Furthermore, plasma membrane integrity and SOD activity were found to be higher (P<0.05) in group supplemented with 0.25 and 0.5% sericin (P<0.05). The MDA concentration was found to be significantly lower (P<0.05) in 0.25 and 0.5% sericin treated groups than control and other treated groups. In conclusion, the supplementation of 0.25-0.5% sericin in semen extender improves frozen-thawed semen quality through protecting sperm from oxidative stress.


Assuntos
Criopreservação/veterinária , Crioprotetores/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Preservação do Sêmen/veterinária , Sêmen/efeitos dos fármacos , Sericinas/farmacologia , Animais , Búfalos , Criopreservação/métodos , Masculino , Sêmen/fisiologia , Preservação do Sêmen/métodos
8.
Ann Med Health Sci Res ; 3(1): 96-8, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23634338

RESUMO

The publication process is a shared responsibility. Besides the writing, reviewing, publishing, and editorial teams, readers are one of the most important pillars of this process. Readers and authors cannot be dealt with separately, because most of the readers are authors. The varieties of articles and improvement in presentations reflect the rising interest and enthusiasm of writers and readers. Increasing number in critical comments and author's reply can be considered as a post-publication peer review process. Impact Factor, which was used as a proxy for the relative importance of a journal, is now being considered a misleading tool in assessing the quality of a paper or the researcher. Here, we are trying to discuss in brief the points which should be kept in mind before manuscript preparation and submission, so that our research should reach to maximum readers in an unbiased form.

9.
J Postgrad Med ; 57(4): 321-8, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22120862

RESUMO

Statins are the most effective and widely used drugs for treating dyslipidemia, a major risk factor for coronary heart disease. These are one of the safest hypolipidemic drugs but many patients are bound to discontinue statins due to their side effects. Hepatotoxicity, myotoxicity and peripheral neuropathy are important out of them. Discontinuation of statins leads to dylipidemia and its grave consequences. Hence, there should be enough strategies for statin intolerant patients, so that they can be saved from these consequences. These side effects can be avoided by the awareness of certain factors viz. potential drug interactions and dose adjustment according to patho-physiology of the patient. Baseline investigations for liver function and muscle toxicity should be done before initiating statin therapy. Here, we are discussing various options for statin intolerant hyperlipidemic patients such as lower and intermittent dosing of statins, alternate hypolipidemic drugs, red yeast rice, supplementation with coenzyme Q10 and vitamin D. A number of hypolipidemic drugs are in trial phases and hold promise for statin intolerant patients.


Assuntos
Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Inibidores de Hidroximetilglutaril-CoA Redutases/efeitos adversos , Hiperlipidemias/tratamento farmacológico , Anticolesterolemiantes/uso terapêutico , Doença Hepática Induzida por Substâncias e Drogas/terapia , Suplementos Nutricionais , Ácidos Fíbricos , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/administração & dosagem , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Estilo de Vida , Doenças Musculares/induzido quimicamente , Doenças Musculares/terapia , Niacina , Ubiquinona/análogos & derivados , Vitamina D
11.
Chest ; 120(6): 1989-97, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11742933

RESUMO

STUDY OBJECTIVE: To describe the clinical course, complications, and prognostic factors of morbidly obese patients admitted to the ICU compared to a control group of nonobese patients. DESIGN: A retrospective study. SETTING: Two university-affiliated hospitals. METHODS: We reviewed the medical records of 117 morbidly obese patients (body mass index >/= 40 kg/m(2)) admitted to the medical ICU between January 1994 and June 2000. Data collected included demographic information, comorbid condition, APACHE (acute physiology and chronic health evaluation) II score, invasive procedures, organ failure, and in-hospital mortality. RESULTS: Obstructive airway disease, pneumonia, and sepsis were the main reasons for admission to the ICU in the morbidly obese group. Sixty-one percent of the morbidly obese patients and 46% of the nonobese group required mechanical ventilation (p = 0.02). The mean lengths of mechanical ventilation and ICU stay were significantly longer for the morbidly obese group (7.7 +/- 9.6 days and 9.3 +/- 10.5 days vs 4.6 +/- 7.1 days and 5.8 +/- 8.2 days, respectively; p < 0.001). APACHE II scores were not significantly different in the two groups (19.1 +/- 7.6 and 20.6 +/- 12.2; p = 0.6). Overall mortality was 30% for the morbidly obese patients and 17% for the nonobese group (p = 0.019). By multivariate analysis, multiorgan failure (odds ratio [OR], 4.6; 95% confidence interval [CI], 2.1 to 16.6), PaO(2)/fraction of inspired oxygen < 200 for > 48 h (OR, 2.3; 95% CI, 1.2 to 7.8), and depressed left ventricular ejection fraction < 40% (OR, 1.4; 95% CI, 1.03 to 13.8) were independently associated with ICU mortality in the morbidly obese group. CONCLUSION: We conclude that critically ill morbidly obese patients are at increased risk of morbidity and mortality compared to the nonobese patients.


Assuntos
Unidades de Terapia Intensiva/estatística & dados numéricos , Obesidade Mórbida/mortalidade , APACHE , Adulto , Idoso , Causas de Morte , Feminino , Mortalidade Hospitalar , Hospitais Universitários , Humanos , Tempo de Internação/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Insuficiência de Múltiplos Órgãos/mortalidade , New York/epidemiologia , Obesidade Mórbida/complicações , Admissão do Paciente/estatística & dados numéricos , Pneumonia/mortalidade , Doença Pulmonar Obstrutiva Crônica/mortalidade , Medição de Risco , Choque Séptico/mortalidade
12.
Respir Care ; 46(7): 702-4, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11403702

RESUMO

Granular cell tumor (GCT) of the lung is a rare tumor, constituting less than 10% of all GCTs. It is a multicentric tumor and infiltrates into adjoining tissue, but malignant GCT of the lung has not been reported. Diagnosis is usually obtained with bronchoscopic biopsy. Treatment options include bronchoscopic extirpation, laser therapy, and sleeve resection. We present a case of GCT co-existing with adenocarcinoma of the lung and review the literature.


Assuntos
Adenocarcinoma Bronquioloalveolar/complicações , Neoplasias Pulmonares/complicações , Granuloma de Células Plasmáticas Pulmonar/complicações , Adulto , Feminino , Humanos
13.
J Hered ; 92(1): 16-22, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11336224

RESUMO

Little is known about the genetics controlling the rate of embryonic development in salmonids, despite the fact that this trait plays an important role in the life history of wild and cultured stocks. We investigated the genetics of embryonic development rate by performing an analysis of quantitative trait loci (QTL) on two families of androgenetically derived doubled haploid rainbow trout produced from a hybrid of two clonal lines with divergent embryonic development rates. A total of 170 doubled haploid individuals were genotyped at 222 marker loci [219 amplified fragment length polymorphism (AFLP) markers, 2 microsatellites, and p53]. A genetic linkage analysis resulted in a map consisting of 27 linkage groups with 21 of the markers remaining unlinked at a minimum LOD of 3.0 and maximum theta of 0.40. Eight of these linkage groups were matched to published rainbow trout linkage groups. Composite interval mapping (CIM) revealed evidence for two QTL influencing time to hatch, and suggestive evidence for a third. These QTL accounted for a total of 24.6% of the variation in time to hatch. One of these QTL had a large effect on development rate, especially in one family of doubled haploids, in which it explained 25.6% of the variance in time to hatch. QTL influencing embryonic length and weight at the commencement of exogenous feeding were also identified. The QTL with the strongest effect on embryonic length (lenR13) mapped to the same position as the QTL with the strongest effect on time to hatch (tthR13), suggesting a single QTL may have a pleiotropic effect on both these traits. These results suggest that the use of clonal lines with a doubled haploid crossing design is an effective way of analyzing the genetic basis of complex traits in salmonids.


Assuntos
Mapeamento Cromossômico , Peso Fetal/genética , Oncorhynchus mykiss/genética , Característica Quantitativa Herdável , Animais , Feminino , Peso Fetal/fisiologia , Genes Dominantes , Escore Lod , Masculino , Repetições de Microssatélites , Polimorfismo de Fragmento de Restrição
14.
Am J Respir Crit Care Med ; 163(3 Pt 1): 645-51, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11254518

RESUMO

The etiology of severe pneumonia requiring mechanical ventilation in the very elderly has been imprecise because of lack of comprehensive studies and low yield of diagnostic approach. Overall, 104 patients 75 yr of age and older with severe pneumonia were studied prospectively at two university-affiliated hospitals. Microbial investigation included blood culture, serology, pleural fluid, and bronchoalveolar secretions. Streptococcus pneumoniae (14%), gram-negative enteric bacilli (14%), Legionella sp. (9%), Hemophilus influenzae (7%), and Staphylococcus aureus (7%) were the predominant pathogens in community-acquired pneumonia (CAP). Staphylococcus aureus (29%), gram-negative enteric bacilli (15%), Streptococcus pneumoniae (9%), and Pseudomonas aeruginosa (4%) accounted for most isolates of nursing home-acquired pneumonia (NHAP). The case fatality rate was 55% (53% for CAP and 57% for NHAP; p > 0.5). Activity of Daily Living (ADL) Index, pulmonary, endocrine and central nervous system (CNS) comorbidities were associated with distinct microbial etiology. By multivariate analysis, hospital mortality was associated independently with 24-h urine output (odds ratio [OR], 5.6; 95% confidence interval [CI], 2.5 to 7.9; p < 0.001), septic shock (OR, 4.3; 95% CI, 1.9 to 8.9; p = 0.0059), radiographic multilobar involvement (OR, 3.7; 95% CI, 1.8 to 15.6; p = 0.02), and inadequate antimicrobial therapy (OR, 2.6; 95% CI, 1.4 to 23.9; p = 0.034). Further studies should focus on identifying effective antimicrobial regimens in randomized trials.


Assuntos
Pneumonia Bacteriana/epidemiologia , Pneumonia Bacteriana/microbiologia , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pneumonia Bacteriana/complicações , Pneumonia Bacteriana/diagnóstico , Pneumonia Bacteriana/tratamento farmacológico , Prevalência , Prognóstico , Estudos Prospectivos , Índice de Gravidade de Doença
15.
J Am Geriatr Soc ; 49(12): 1614-21, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11843993

RESUMO

OBJECTIVES: To develop a prognostic model to predict outcome of older patients with severe pneumonia requiring mechanical ventilation. DESIGN: A nonconcurrent prospective study. SETTING: A 24-bed intensive care unit (ICU) within two university-affiliated tertiary care hospitals. PARTICIPANTS: All patients age 75 and older with severe pneumonia between June 1996 and September 1999 were included. Demographic data including activities of daily living (ADL) index score before admission, and clinical and laboratory data were collected in the first 24 hours of admission to the ICU. One hundred four patients (mean age +/- standard deviation (SD) 82.3 +/- 5.5 years) met the inclusion criteria. MEASUREMENTS: A classification tree was developed using binary recursive partitioning to predict hospital discharge. The model was compared with a logistic regression model using variables selected by the tree analysis and with the Acute Physiologic and Chronic Health Evaluation (APACHE) II. RESULTS: Outcome predictors for the classification tree were use of vasopressors, presence of multilobar pneumonia on chest radiograph, ratio of blood urea nitrogen to creatinine, Glasgow Coma Scale, urine output, and ADL score before admission. The tree achieved a sensitivity of 83.8% (95% confidence interval (CI) 69.2-92.4) and a specificity of 93.3% (95% CI 83-98.1). The predictive accuracy as assessed by the area under the curve (c-index +/- standard error) was significantly higher with the classification tree (0.932 +/- 0.03) than with logistic regression and APACHE II, (0.801 +/- 0.028 and 0.711 +/- 0.049, respectively (P < .05). CONCLUSIONS: The classification tree model demonstrated a superior predictive accuracy to that of logistic regression and APACHE II. If validated prospectively, the classification tree can be used as a tool to assess the outcome of older patients with severe pneumonia requiring mechanical ventilation on admission to the ICU. In addition, the classification tree can be used to assist healthcare workers in providing a concise summary of local outcome experience and prognostic information to patients and their surrogates.


Assuntos
Modelos Estatísticos , Avaliação de Resultados em Cuidados de Saúde , Pneumonia/terapia , Respiração Artificial , APACHE , Atividades Cotidianas , Idoso , Idoso de 80 Anos ou mais , Estudos de Coortes , Feminino , Hospitais Universitários , Humanos , Unidades de Terapia Intensiva , Masculino , Pneumonia/mortalidade , Valor Preditivo dos Testes , Prognóstico , Estudos Prospectivos , Análise de Regressão , Sensibilidade e Especificidade , Índice de Gravidade de Doença
17.
Intensive Care Med ; 26(12): 1803-10, 2000 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11271088

RESUMO

OBJECTIVE: To evaluate the predictive ability of three severity of illness scoring systems in elderly patients with severe pneumonia requiring mechanical ventilation compared to a younger age group. DESIGN: Prospective cohort study. SETTING: Two university-affiliated tertiary care hospitals. PATIENTS AND PARTICIPANTS: One hundred four patients 75 years of age and older and 253 patients younger than 75 years of age enrolled from medical intensive care units. MEASUREMENTS AND RESULTS: Probabilities of hospital death for patients were estimated by the Acute Physiology and Chronic Health Evaluation (APACHE) II, the Mortality Probability Model (MPM) II and the Simplified Acute Physiology Score (SAPS) II. Predicted risks of hospital death were compared with observed outcomes using three methods of assessing the overall goodness of fit. The actual mortality of the elderly group was 54.87 % (95 % confidence interval [CI]: 45.2-64.4 %) compared to 28.9 % (95 % CI, 23.3-34.4 %) in the younger age group. There was a significant difference in the predictive accuracy of the scoring systems as assessed by the c-index, which is equivalent to the area under the receiver operator characteristics (ROC) curve, between the two groups, but not within individual groups. Calibration was insufficient for APACHE II and SAPS II in the elderly cohort as in-hospital mortality was lower than the predicted mortality for both models. CONCLUSIONS: Although the three severity of illness scoring systems (APACHE II, MPM II and SAPS II) demonstrated average discrimination when applied to estimate hospital mortality in the elderly patients with severe pneumonia, MPM II had the closest fit to our database. Alternative modeling approaches might be needed to customize the model coefficients to the elderly population for more accurate probabilities or to develop specialized models targeted to the designed population.


Assuntos
APACHE , Idoso/estatística & dados numéricos , Mortalidade Hospitalar , Pneumonia/classificação , Pneumonia/mortalidade , Índice de Gravidade de Doença , Idoso de 80 Anos ou mais , Calibragem , Cuidados Críticos/normas , Análise Discriminante , Feminino , Hospitais Universitários , Humanos , Unidades de Terapia Intensiva/estatística & dados numéricos , Tempo de Internação/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Modelos Estatísticos , New York/epidemiologia , Avaliação de Resultados em Cuidados de Saúde , Pneumonia/diagnóstico , Pneumonia/terapia , Estudos Prospectivos , Respiração Artificial , Fatores de Risco , Sensibilidade e Especificidade , Fatores de Tempo
18.
Chem Biol Interact ; 114(1-2): 15-31, 1998 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-9744553

RESUMO

Deficiency of the vitamin folic acid has recently been linked with increased incidence of neural tube defects and of cardiovascular disease, through elevated plasma homocysteine levels. The kidney has an important role in conserving folate to counteract development of deficiency. Urinary folate excretion is regulated by the degree of reabsorption of folate by the proximal tubule cell. To evaluate an in vitro model for studies of the regulation of urinary folate excretion, the present studies examined the transport of 5-methyltetrahydrofolate (5-CH3-H4PteGlu), the primary form of folate in the glomerular filtrate, by normal rat proximal tubule (RPT) cells in confluent monolayer cultures. Specific binding of 5-CH3-H4PteGlu to the apical membrane was saturable (K(D) = 27 nM), but intracellular transport was not saturated up to 100 nM concentrations. 5-CH3-H4PteGlu transport was decreased 50% by concentrations of folic acid that completely blocked 5-CH3-H4PteGlu binding by the apical folate receptor. Probenecid (10 mM), an anion exchange (reduced folate carrier) inhibitor, reduced 5CH3-H4PteGlu transport by 50% without significantly affecting binding. Aspirin (3 mM) did not alter 5-CH3-H4PteGlu transport, but significantly enhanced the inhibition due to probenecid. Similarly, indomethacin (5 microM) potentiated the inhibition of 5-CH3-H4PteGlu transport by probenecid. These data suggest that RPT cells take up 5-CH3-H4PteGlu by both the folate receptor and the reduced folate carrier, implying a role for both pathways in regulating urinary folate excretion.


Assuntos
Proteínas de Transporte/metabolismo , Túbulos Renais Proximais/metabolismo , Proteínas de Membrana , Proteínas de Membrana Transportadoras , Receptores de Superfície Celular , Tetra-Hidrofolatos/metabolismo , Animais , Proteínas de Transporte de Ânions , Aspirina/farmacologia , Transporte Biológico , Proteínas de Transporte/urina , Células Cultivadas , Receptores de Folato com Âncoras de GPI , Ácido Fólico/análogos & derivados , Ácido Fólico/metabolismo , Taxa de Filtração Glomerular , Indometacina/farmacologia , Masculino , Probenecid/farmacologia , Ratos , Ratos Sprague-Dawley , Proteína Carregadora de Folato Reduzido , Fármacos Renais/farmacologia , Tetra-Hidrofolatos/urina
19.
In Vitro Cell Dev Biol Anim ; 32(5): 285-91, 1996 May.
Artigo em Inglês | MEDLINE | ID: mdl-8792158

RESUMO

An in vitro model to establish primary and subcultures of rat kidney proximal tubule (RPT) cells is described. After excising the kidneys and separating the cortex, the cortical tissue is digested with the enzymes DNAse-collagenase (Type I) resulting in a high yield of viable RPT Cells. The isolated RPT cells are then seeded onto rat tail collagen-coated surfaces and grown to confluency in a serum-free, hormonally defined medium. The cell yield can be increased by transferring the conditioned medium on Day 1 to more rat tail collagen-coated surfaces. RPT cell attachment and morphology was better on rat tail collagen-coated surfaces than on bovine collagen Type I coated surfaces. The culture medium was a 1:1 mixture of Ham's F-12 and Dulbecco's modified Eagle's medium supplemented with bovine serum albumin, insulin, transferrin, selenium, hydrocortisone, triiodothyronine, epidermal growth factor, and glutamine. The RPT cells became confluent in 7-10 d, at which point they could be subcultured by trypsinizing and growth in the same medium. In some studies, 10 ng/ml cholera toxin was added to the culture medium. We could passage the RPT cells up to 14 times in the presence of cholera toxin. The cells were investigated for activity of several markers. The cells were histochemically positive for alkaline phosphatase and gamma-glutamyl transpeptidase activity and synthesized the intermediate filament pankeratin. The RPT cells displayed apically directed sodium-dependent active glucose transport in culture. Hence, the RPT cells retain structural and functional characteristics of transporting renal epithelia in culture. This rat cell culture model will be a valuable tool for substrate uptake and nephrotoxicity studies.


Assuntos
Túbulos Renais Proximais/citologia , Animais , Transporte Biológico , Divisão Celular , Células Cultivadas , Colágeno , Meios de Cultivo Condicionados , Meios de Cultura Livres de Soro , Matriz Extracelular , Glucose/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley
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