RESUMO
Choline and methionine may serve unique functions to alter hepatic energy metabolism. Our objective was to trace carbon flux through pathways of oxidation and glucose metabolism in bovine hepatocytes exposed to increasing concentrations of choline chloride (CC) and D,L-methionine (DLM). Primary hepatocytes were isolated from 4 Holstein calves and maintained for 24 h before treatment with CC (0, 10, 100, 1000 µmol/L) and DLM (0, 100, 300 µmol/L) in a factorial design. After 21 h, [1-14C]C16:0 or [2-14C]pyruvate was added to measure complete and incomplete oxidation, and cellular glycogen. Reactive oxygen species (ROS), cellular triglyceride (TG), and glucose and ß-hydroxybutyrate (BHB) export were quantified. Exported very-low density lipoprotein particles were isolated for untargeted lipidomics and to quantify TG. Interactions between CC and DLM, and contrasts for CC (0 vs. [10, 100, 1000 µmol/L] and linear and quadratic contrast 10, 100, 1000 µmol/L) and DLM (0 vs. [100, 300 µmol/L] and 100 vs. 300 µmol/L) were evaluated. Presence of CC increased complete oxidation of [1-14C]C16:0 and decreased BHB export. Glucose export was decreased, but cellular glycogen was increased by the presence of CC and increasing CC. Presence of CC decreased ROS and marginally decreased cellular TG. No interactions between CC and DLM were detected for these outcomes. These data suggest a hepato-protective role for CC to limit ROS and cellular TG accumulation, and to alter hepatic energy metabolism to support complete oxidation of FA and glycogen storage regardless of Met supply.
Assuntos
Bovinos/metabolismo , Hepatócitos/metabolismo , Palmitatos/metabolismo , Ácido Pirúvico/metabolismo , Animais , Animais Recém-Nascidos , Ciclo do Carbono/efeitos dos fármacos , Células Cultivadas , Colina/metabolismo , Colina/farmacologia , Metabolismo Energético/efeitos dos fármacos , Ácidos Graxos/metabolismo , Glucose/metabolismo , Hepatócitos/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Metionina/metabolismo , Metionina/farmacologia , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Triglicerídeos/metabolismoRESUMO
Nutritional interventions, either by controlling dietary energy (DE) or supplementing rumen-protected choline (RPC) or both, may mitigate negative postpartum metabolic health outcomes. A companion paper previously reported the effects of DE density and RPC supplementation on production and health outcomes. The objective of this study was to examine the effects of DE and RPC supplementation on the expression of hepatic oxidative, gluconeogenic, and lipid transport genes during the periparturient period. At 47 ± 6 d relative to calving (DRTC), 93 multiparous Holstein cows were randomly assigned in groups to dietary treatments in a 2 × 2 factorial of (1) excess energy (EXE) without RPC supplementation (1.63 Mcal of NEL/kg of dry matter; EXE-RPC); (2) maintenance energy (MNE) without RPC supplementation (1.40 Mcal of NEL/kg dry matter; MNE-RPC); (3) EXE with RPC supplementation (EXE+RPC); and (4) MNE with RPC supplementation (MNE+RPC). To achieve the objective of this research, liver biopsy samples were collected at -14, +7, +14, and +21 DRTC and analyzed for mRNA expression (n = 16/treatment). The interaction of DE × RPC decreased glucose-6-phosphatase and increased peroxisome proliferator-activated receptor α in MNE+RPC cows. Expression of cytosolic phosphoenolpyruvate carboxykinase was altered by the interaction of dietary treatments with reduced expression in EXE+RPC cows. A dietary treatment interaction was detected for expression of pyruvate carboxylase although means were not separated. Dietary treatment interactions did not alter expression of carnitine palmitoyltransferase 1A or microsomal triglyceride transfer protein. The 3-way interaction of DE × RPC × DRTC affected expression of carnitine palmitoyltransferase 1A, glucose-6-phosphatase, and peroxisome proliferator-activated receptor α and tended to affect cytosolic phosphoenolpyruvate carboxykinase. Despite previously reported independent effects of DE and RPC on production variables, treatments interacted to influence hepatic metabolism through altered gene expression.
Assuntos
Bovinos/genética , Colina/administração & dosagem , Ingestão de Energia/fisiologia , Gluconeogênese/genética , Metabolismo dos Lipídeos/genética , Fígado/metabolismo , Animais , Bovinos/fisiologia , Dieta/veterinária , Suplementos Nutricionais , Feminino , Expressão Gênica/fisiologia , Glucose-6-Fosfatase/metabolismo , Lactação/efeitos dos fármacos , Leite/metabolismo , Período Periparto/efeitos dos fármacos , Gravidez , Cuidado Pré-Natal , Rúmen/metabolismoRESUMO
Our previously published paper demonstrated that fermented ammoniated condensed whey (FACW) supplementation improved feed efficiency and metabolic profile in postpartum dairy cows. The objective of this study was to further explore the effects of FACW supplementation on liver triglyceride content, hepatic gene expression and protein abundance, and plasma biomarkers related to liver function, inflammation, and damage. Individually fed multiparous Holstein cows were blocked by calving date and randomly assigned to postpartum (1 to 45 d in milk, DIM) isonitrogenous treatments: control diet (n = 20) or diet supplemented with FACW (2.9% dry matter of diet as GlucoBoost; Fermented Nutrition, Luxemburg, WI, replacing soybean meal; n = 19). Liver biopsies were performed at 14 and 28 DIM for analysis of mRNA expression, protein abundance, and liver triglyceride content. There was marginal evidence for a reduction in liver triglyceride content at 14 DIM in FACW-supplemented cows compared with the control group. Cows supplemented with FACW had greater mRNA expression of glucose-6-phosphatase at 14 DIM relative to control. Supplementation with FACW increased mRNA expression of pyruvate carboxylase (PC), but did not alter cytosolic phosphoenolpyruvate carboxykinase (PCK1), resulting in a 2.4-fold greater PC:PCK1 ratio for FACW-supplemented cows compared with control. There was no evidence for a FACW effect on mRNA expression of propionyl-CoA carboxylase nor on mRNA expression or protein abundance of lactate dehydrogenase A or B. Cows supplemented with FACW had lower plasma urea nitrogen compared with control. Plasma l-lactate was greater for FACW-supplemented cows compared with control at 2 h before feeding time at 21 DIM. There was no evidence for altered expression of IL1B or IL10, or blood biomarkers related to liver function and damage. Greater glucose-6-phosphatase and PC gene expression, together with greater blood glucose and similar milk lactose output, suggests that FACW increased the supply of glucose precursors, resulting in greater gluconeogenesis between 3 and 14 DIM. Greater hepatic PC:PCK1 ratio, together with previously reported decreased plasma ß-hydroxybutyrate and the marginal evidence for lower liver triglyceride content at 14 DIM, suggests greater hepatic capacity for complete oxidation of fatty acids in FACW-supplemented cows compared with control. Overall, improvements in metabolite profile and feed efficiency observed with postpartum supplementation of FACW may be attributed to increased gluconeogenic and anaplerotic precursors, most likely propionate, due to modulated rumen fermentation.
Assuntos
Bovinos/fisiologia , Suplementos Nutricionais/análise , Leite/metabolismo , Soro do Leite/administração & dosagem , Ácido 3-Hidroxibutírico/sangue , Compostos de Amônio/química , Animais , Dieta/veterinária , Feminino , Fermentação , Gluconeogênese/efeitos dos fármacos , Glucose/metabolismo , Glucose-6-Fosfatase/genética , Lactação/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Nutrientes/metabolismo , Período Pós-Parto/efeitos dos fármacos , Piruvato Carboxilase/genética , Distribuição Aleatória , Rúmen/metabolismoRESUMO
During the peripartum period, dairy cows experience both an increase in circulating fatty acid (FA) profile and a change in circulating FA profile, which have been shown to alter regulation of gluconeogenic genes. The objective was to quantify gene expression of key enzymes involved in gluconeogenesis and FA transport into the mitochondria in primary hepatocytes in response to exposure to an FA mixture mimicking what is circulating in a transition dairy cow with or without enrichment of C16:0, C18:0, and C18:1. Primary hepatocytes were isolated from 4 Holstein bull calves 3 d of age (± standard deviation 2 d) and cultured. Twenty-four hours after plating, treatments were applied to the cells for 24-h incubation. Treatments consisted of (1) control (1% BSA), (2) 0.75 mM FA cocktail (3% C14:0, 27% C16:0, 23% C18:0, 31% C18:1, 8% C18:2, and 8% C18:3 to mimic the FA profile of dairy cattle at calving), (3) 0.90 mM FA cocktail, (4) 0.75 mM FA cocktail + 0.15 mM C16:0, (5) 0.75 mM FA cocktail + 0.15 mM C18:0, and (6) 0.75 mM FA cocktail + 0.15 mM C18:1. After harvest in Trizol (Life Technologies, Carlsbad, CA), samples were stored at -80°C until RNA extraction, purification, and reverse transcription. Abundance of mRNA was measured using quantitative real-time PCR. Expression of genes of interest [carnitine palmitoyltransferase 1A, pyruvate carboxylase, cytosolic phosphoenolpyruvate carboxykinase (PCK1), mitochondrial phosphoenolpyruvate carboxykinase, and glucose-6-phosphatase] was calculated relative to the average abundance of 2 reference genes (ribosomal protein L32 and glyceraldehyde 3-phosphate dehydrogenase), which were the most stable out of 3 tested. Data were analyzed using PROC MIXED (SAS version 9.4; SAS Institute, Cary, NC) with the fixed effect of treatment and calf in the random statement. The addition of FA compared with the 1% BSA treatment increased the expression of carnitine palmitoyltransferase 1A and cytosolic PCK1. Enrichment with individual FA did not further regulate pyruvate carboxylase or PCK1 beyond that achieved by the basal profile. These results suggest that shifts in circulating FA profile within a biological range, without a difference in the total FA concentration, have minimal effects on transcriptional regulation of hepatic gluconeogenic genes in primary bovine hepatocytes.
Assuntos
Bovinos/metabolismo , Ácidos Graxos/metabolismo , Gluconeogênese , Hepatócitos/metabolismo , Animais , Bovinos/genética , Células Cultivadas , Feminino , Expressão Gênica , Regulação da Expressão Gênica , Glucose-6-Fosfatase/genética , Glucose-6-Fosfatase/metabolismo , Lactação , Fígado/metabolismo , Mitocôndrias/genética , Mitocôndrias/metabolismo , Fosfoenolpiruvato Carboxiquinase (ATP)/genética , Fosfoenolpiruvato Carboxiquinase (ATP)/metabolismo , Piruvato Carboxilase/genética , Piruvato Carboxilase/metabolismoRESUMO
Background: Fournier's gangrene (FG) is a life-threatening infection of the genital, perineal, and perianal regions with a morbidity range between 3 and 67%. Our aim is to report our experience in treatment of FG and to assess whether three different scoring systems can accurately predict mortality and morbidity in FG patients. Methods: All patients that were treated for FG at the Department of Urology of the University Hospital Basel between June 2012 and March 2017 were included and assessed retrospectively by chart review. Furthermore, we calculated Fournier's Gangrene Severity Index (FGSI), the Laboratory Risk Indicator for Necrotizing Fasciitis (LRINEC), and the neutrophil-lymphocyte ratio (NLR) in every patient and assessed whether those scores correlate with the patients' morbidity and mortality. Results: Twenty patients were included, with a median (IQR) age of 66 (46-73) years. Fifteen of twenty (75%) patients required treatment on an intensive care unit, and three died (mortality rate: 15%). The mean FGSI, LRINEC, and NLR scores were 13.0, 9.3, and 45.3 for non-survivors and 7.7, 6.5, and 26 for survivors, respectively. None of the risk scores correlated significantly with mortality; however, all three significantly correlated with infection- and surgically-induced morbidity. Conclusions: In our series, Fournier's gangrene was associated with a mortality rate of 15% despite maximum multidisciplinary therapy at a specialized center. All risk scores were able to predict the morbidity of the disease in terms of local extent and the required surgical measures.
Assuntos
Gangrena de Fournier/classificação , Gangrena de Fournier/mortalidade , Índice de Gravidade de Doença , Idoso , Feminino , Gangrena de Fournier/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Estatísticas não Paramétricas , Suíça/epidemiologia , Centros de Atenção Terciária/organização & administração , Centros de Atenção Terciária/estatística & dados numéricosRESUMO
To date, osteometric data for Swiss Bronze Age cattle, particularly from Alpine sites, are scarce. In the present study, using a large dataset generated by combining preexisting data with recent data obtained from a large Alpine site, cattle size from the Late Neolithic to the Late Bronze Age (LBA) in populations from different sites and regions was evaluated using the logarithmic size index and other statistical analysis. Additionally, the finite mixture model and a meta-analytic technique were used to observe possible effects of sex ratios on cattle size. Results indicated that sex ratios did not affect size distribution. Cattle populations did not differ over time, but the Alpine cattle were smaller than the Central Plateau cattle. There were two distinct sizes in the Alpine cattle populations. It is suggested that the different economic interrelationships between Alpine and other geographically related communities might have led to the emergence of size diversity in Swiss Bronze Age cattle. Further interdisciplinary studies with larger sample sizes are required to confirm these possibilities.
RESUMO
Accurate cow-side blood ß-hydroxybutyrate (BHB) detection meters are valuable tools for rapid diagnosis of hyperketonemia. The main objective of this study was to compare the blood BHB measured in whole blood by the BHBCheck meter (PortaCheck, Moorestown, NJ) to a previously validated meter, Precision Xtra meter (Abbott Laboratories, Abbott Park, IL) and a colorimetric laboratory assay. Samples (n = 426) were collected from postpartum primiparous and multiparous Holstein cows (n = 79 cows) enrolled in 1 of 2 experiments (Exp) with different sampling schedules (Exp 1: n = 39 cows, 58 samples; Exp 2: n = 40 cows, 368 samples). In both Exp, whole-blood samples were collected from the coccygeal vessels after morning milking, before morning feeding. Blood samples were used immediately for BHB quantification via the BHBCheck meter and the Precision Xtra meter. Blood was also collected into evacuated tubes containing no additive (Exp 1) or potassium oxalate/sodium fluoride (Exp 2), which were centrifuged for serum or plasma separation and stored at -20°C for subsequent analysis. Laboratory quantification of BHB concentration was done by the BHB LiquiColor Assay (EKF Diagnostics-Stanbio, Boerne, TX; certified for serum and plasma). Data were analyzed by UNIVARIATE, CORR, FREQ, REG, and LOGISTIC procedures of SAS 9.4 (SAS Institute Inc., Cary, NC). Within this sample set, average parity was 3.3 lactations and DIM was 14 d. The proportion of samples classified as hyperketonemia (BHB ≥1.2 mmol/L) was 25, 28, and 31% as determined by the colorimetric assay, BHBCheck meter, and Precision Xtra meter, respectively. The correlation for BHBCheck meter BHB concentration compared with the colorimetric assay concentrations was r = 0.96, with a sensitivity of 91% and specificity of 93%. Correlation, sensitivity, and specificity of the Precision Xtra meter concentrations were 0.97, 98%, and 92%, respectively. Bland-Altman plots demonstrated minimal bias for both meters. Area under the receiver operator characteristic curve suggests adequate diagnostic accuracy of both meters. Overall, accuracy, sensitivity, and specificity of the BHBCheck meter was similar to the Precision Xtra meter and laboratory assay, indicating the BHBCheck meter is appropriate for use as a cow-side diagnostic test for hyperketonemia in dairy cows.
Assuntos
Ácido 3-Hidroxibutírico/sangue , Doenças dos Bovinos/diagnóstico , Cetose/veterinária , Sistemas Automatizados de Assistência Junto ao Leito , Animais , Bovinos , Doenças dos Bovinos/sangue , Testes Diagnósticos de Rotina/instrumentação , Testes Diagnósticos de Rotina/métodos , Testes Diagnósticos de Rotina/veterinária , Feminino , Cetose/sangue , Cetose/diagnóstico , Paridade , Gravidez , Curva ROC , Sensibilidade e EspecificidadeRESUMO
For therapeutic drug monitoring in remote settings, dried blood spots (DBS) are particularly advantageous, as blood sample collection and handling is uncomplicated. The aim of this study was to develop and validate an automated extraction method for the analysis of nevirapine, efavirenz and lopinavir in DBS samples. Automated extraction was performed with methanol : water (70 : 30 v/v), using a DBS-MS 500 autosampler coupled to a liquid chromatography tandem mass spectrometry system. The autosampler used digital images of each DBS to position the extraction head, sprayed 10 µl of internal standard onto each DBS and extracted a 4-mm disc (Ø) from the centre of each spot by unilateral flow using 25-µl extraction solvent. The analytes were baseline separated on a pentafluorophenyl column and analysed by using electrospray ionization with multiple reaction monitoring in positive polarity mode for nevirapine and lopinavir and in negative mode for efavirenz. The method was linear between 10 and 10 000 ng/ml for all analytes. Automated sample extraction resulted in consistent recoveries (nevirapine: 70 ± 6%, efavirenz: 63 ± 11% and lopinavir: 60 ± 10%) and matrix effects between different donors and concentration levels. Intra-day and inter-day accuracy and precision deviations were ≤15%. Manual and automated extractions of DBS samples collected within the framework of an adherence assessment study in rural Tanzania showed good agreements with deviations of less than 10%. Our study highlights that therapeutic drug monitoring samples obtained in the resource-constrained setting of rural Africa can be reliably determined by automated extraction of DBS. Overall, automatization improved method sensitivity and facilitates analysis of large sample numbers. Copyright © 2017 John Wiley & Sons, Ltd.
Assuntos
Antirretrovirais/sangue , Teste em Amostras de Sangue Seco/métodos , Ensaios de Triagem em Larga Escala/métodos , Alcinos , Benzoxazinas/análise , Cromatografia Líquida de Alta Pressão , Ciclopropanos , Humanos , Limite de Detecção , Lopinavir/análise , Nevirapina/análise , Sensibilidade e Especificidade , Espectrometria de Massas em TandemRESUMO
OBJECTIVES: Self-reported adherence assessment in HIV-infected patients on antiretroviral therapy (ART) is challenging and may overestimate adherence. The aim of this study was to improve the ability of health care providers to elicit patients' reports of nonadherence using a "patient-centred" approach in a rural sub-Saharan African setting. METHODS: A prospective interventional cohort study of HIV-infected patients on ART for ≥ 6 months attending an HIV clinic in rural Tanzania was carried out. The intervention consisted of a 2-day workshop for health care providers on patient-centred communication and the provision of an adherence assessment checklist for use in the consultations. Patients' self-reports of nonadherence (≥ 1 missed ART dose/4 weeks), subtherapeutic plasma ART concentrations (< 2.5th percentile of published population-based pharmacokinetic models), and virological and immunological failure according to the World Health Organization definition were assessed before and after (1-3 and 6-9 months after) the intervention. RESULTS: Before the intervention, only 3.3% of 299 patients included in the study reported nonadherence. Subtherapeutic plasma ART drug concentrations and virological and immunological failure were recorded in 6.5%, 7.7% and 14.5% of the patients, respectively. Two months after the intervention, health care providers detected significantly more patients reporting nonadherence compared with baseline (10.7 vs. 3.3%, respectively; P < 0.001), decreasing to 5.7% after 6-9 months. A time trend towards higher drug concentrations was observed for efavirenz but not for other drugs. The virological failure rate remained unchanged whereas the immunological failure rate decreased from 14.4 to 8.7% at the last visit (P = 0.002). CONCLUSIONS: Patient-centred communication can successfully be implemented with a simple intervention in rural Africa. It increases the likelihood of HIV-infected patients reporting problems with adherence to ART; however, sustainability remains a challenge.
Assuntos
Fármacos Anti-HIV/uso terapêutico , Infecções por HIV/tratamento farmacológico , Pessoal de Saúde/educação , Adulto , Lista de Checagem , Feminino , Humanos , Masculino , Adesão à Medicação , Pessoa de Meia-Idade , Assistência Centrada no Paciente , Relações Profissional-Paciente , Estudos Prospectivos , População Rural , Autorrelato , Tanzânia , Resultado do TratamentoRESUMO
The optimal approach in laboratory diagnosis of Clostridium difficile infection (CDI) is still not well defined. Toxigenic culture (TC) or alternatively fecal toxin assay by cell cytotoxicity neutralization assay are considered to be the reference standard, but these methods are time-consuming and labor intensive. In many medical centers, diagnosis of CDI is therefore still based on fecal toxin A/B enzyme immunoassay (EIA) directly from stool alone, balancing cost and speed against limited diagnostic sensitivity. The aim of the study was to assess in which patient population the additional workload of TC is justified. All consecutive stool specimens submitted for diagnosis of suspected CDI between 2004 and 2011 at a tertiary-care center were examined by toxin EIA and TC. Clinical data of patients with established diagnosis of CDI were collected in a standardized case-report form. From 12,481 stool specimens submitted to the microbiologic laboratory, 480 (3.8%) fulfilled CDI criteria; 274 (57.1%) were diagnosed by toxin EIA; and an additional 206 (42.9%) were diagnosed by TC when toxin EIA was negative. Independent predictors for negative toxin EIA but positive TC were high-dose corticosteroids (odds ratio (OR) 2.97, 95% confidence interval (CI) 1.50-5.90, p 0.002), leukocytopenia <1000/µL (OR 2.52, 95% CI 1.22-5.23, p 0.013) and nonsevere CDI (OR 2.21, 95% CI 1.39-3.50, p 0.001). There was no difference in outcomes such as in-hospital mortality and recurrence between both groups. In conclusion, negative toxin EIA does not rule out CDI in immunocompromised patients in the setting of relevant clinical symptoms. Methods with improved sensitivity such as TC or PCR should be used, particularly in this patient population.
Assuntos
Proteínas de Bactérias/análise , Toxinas Bacterianas/análise , Clostridioides difficile/crescimento & desenvolvimento , Infecções por Clostridium/diagnóstico , Enterocolite/diagnóstico , Enterotoxinas/análise , Fezes/química , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Clostridioides difficile/metabolismo , Infecções por Clostridium/microbiologia , Técnicas Citológicas/métodos , Enterocolite/microbiologia , Feminino , Humanos , Hospedeiro Imunocomprometido , Técnicas Imunoenzimáticas/métodos , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Centros de Atenção Terciária , Adulto JovemRESUMO
OBJECTIVE: Despite reduced workplace exposures, beryllium sensitization and chronic beryllium disease still occur. Effective health and safety training is needed. METHODS: Through an Occupational Safety and Health Administration (OSHA) Targeted Topic Training grant and company partners, we developed a training program. Evaluation and validation included knowledge and training reaction assessments and training impact survey. RESULTS: We describe herein the iterative, five-pronged approach: (1) needs assessment; (2) materials development; (3) pilot-testing, evaluation, and material revisions; (4) worker training; and (5) evaluation and validation. Mean posttraining test score increased 14% (82% to 96%; P < 0.005) and were unchanged at 90-day follow-up (94%; P = 0.744). In addition, 49% reported making changes in work practices. CONCLUSIONS: The use of a five-pronged training program was effective and well received and resulted in improved work practices. These materials are available on the OSHA Web site.
Assuntos
Beriliose/prevenção & controle , Serviços de Saúde do Trabalhador/métodos , Saúde Ocupacional/educação , Avaliação Educacional , Humanos , Comunicação Interdisciplinar , Avaliação das Necessidades , Projetos Piloto , Desenvolvimento de Programas , Avaliação de Programas e Projetos de Saúde , Reprodutibilidade dos Testes , Estados Unidos , United States Occupational Safety and Health AdministrationRESUMO
AIM: To establish a refined model of artificially infected root canals and confirm its suitability as a sensitive ex vivo method to assess the efficacy of disinfecting agents. Disinfection was evaluated using sodium hypochlorite (NaOCl), either blocked or unblocked by sodium thiosulphate, and a recently promoted method of disinfection, the antibacterial photodynamic therapy (PDT). METHODOLOGY: The roots of bovine incisors were sectioned into three parts, the canals of coronal and middle regions were filled with a suspension of Enterococcus faecalis and the apical region with culture medium. After 7 days, coronal sections were disinfected using NaOCl (0.5%, 1.0% and 3.0% for 30, 60 and 600 s) or a system for photoactivated chemotherapy (PACT; Cumdente, Tübingen, Germany) for antibacterial PDT. Apical sections served as sterile controls and middle sections as bacterial growth controls. In half of the NaOCl-treated specimens, disinfection was arrested. Dentine chips from biopsies at different depths from the central canal towards the periphery were plated and assessed for colony-forming units (CFU). Disinfection was considered biologically relevant if the reduction of CFU was at least three log10 orders of magnitude. RESULTS: Colony-forming units of 10³ - 104 in growth controls indicated effective artificial infection. A biologically relevant reduction of CFU was accomplished with unblocked NaOCl, but not after blocking with NaOCl nor with PDT. CONCLUSIONS: The system reliably detected disinfection of the root canal and dentinal tubules and proved suitable for ex vivo testing of root canal disinfection. The effect of NaOCl depended on the duration of impact. Under the present experimental conditions, the antibacterial PDT system did not achieve sufficient disinfection.
Assuntos
Cavidade Pulpar/microbiologia , Fotoquimioterapia/métodos , Irrigantes do Canal Radicular/uso terapêutico , Hipoclorito de Sódio/uso terapêutico , Animais , Carga Bacteriana/efeitos dos fármacos , Bovinos , Quelantes/farmacologia , Desinfetantes de Equipamento Odontológico/administração & dosagem , Desinfetantes de Equipamento Odontológico/uso terapêutico , Dentina/efeitos dos fármacos , Dentina/microbiologia , Desinfecção/métodos , Relação Dose-Resposta a Droga , Enterococcus faecalis/efeitos dos fármacos , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Terapia com Luz de Baixa Intensidade , Microscopia Eletrônica de Varredura , Fármacos Fotossensibilizantes/administração & dosagem , Fármacos Fotossensibilizantes/uso terapêutico , Irrigantes do Canal Radicular/administração & dosagem , Hipoclorito de Sódio/administração & dosagem , Tiossulfatos/farmacologia , Técnicas de Cultura de Tecidos , Cloreto de Tolônio/administração & dosagem , Cloreto de Tolônio/uso terapêuticoRESUMO
BACKGROUND AND PURPOSE: The endocannabinoid and corticotropin-releasing factor (CRF) systems have been implicated in several long-lasting behavioural effects of prior cocaine experience. The present experiments were designed to probe functional interactions between endocannabinoids and CRF by testing the role of cannabinoid CB(1) receptors in cocaine-related behaviours induced or mediated by CRF. EXPERIMENTAL APPROACH: In Experiment 1, rats trained to self-administer cocaine were pretreated with the CB(1) receptor antagonist, AM251 (0, 10, 100 or 200 µg, i.c.v.), before tests for reinstatement in response to CRF (0, 0.5 µg, i.c.v.), intermittent footshock stress (0, 0.9 mA) or cocaine (0, 10 mg·kg(-1) , i.p.). In Experiment 2, rats pre-exposed to cocaine (15-30 mg·kg(-1) , i.p.) or saline for 7 days were pretreated with AM251 (0, 10 or 100 µg, i.c.v.) before tests for locomotion in response to CRF (0.5 µg, i.c.v.), cocaine (15 mg·kg(-1) , i.p.) or saline (i.c.v.). KEY RESULTS: Pretreatment with AM251 selectively interfered with CRF-, but not footshock- or cocaine-induced reinstatement. AM251 blocked the expression of behavioural sensitization induced by challenge injections of both CRF and cocaine. CONCLUSIONS AND IMPLICATIONS: These findings reveal a mediating role for CB(1) receptor transmission in the effects of CRF on cocaine-related behaviours.
Assuntos
Transtornos Relacionados ao Uso de Cocaína/fisiopatologia , Hormônio Liberador da Corticotropina/farmacologia , Receptor CB1 de Canabinoide/fisiologia , Animais , Comportamento Animal/efeitos dos fármacos , Antagonistas de Receptores de Canabinoides/farmacologia , Cocaína/administração & dosagem , Eletrochoque , Masculino , Atividade Motora/efeitos dos fármacos , Piperidinas/farmacologia , Pirazóis/farmacologia , Ratos , Ratos Long-Evans , Ratos Wistar , Receptor CB1 de Canabinoide/antagonistas & inibidores , Autoadministração , Estresse FisiológicoRESUMO
The endocannabinoid (eCB) system is an important regulator of the stress response and mediates several stress-related behaviors, including anxiety. Despite anatomical evidence that eCBs interact with the principle stress peptide, corticotropin-releasing factor (CRF), few data exist that address functional interactions between these systems. Accordingly, we examined the effects of the CB1 receptor antagonist, AM251, on behavioral anxiety induced by (1) exogenous CRF, and (2) withdrawal from chronic cocaine exposure (mediated by CRF). After behavioral testing, we collected blood and assessed plasma corticosterone levels. In Experiment 1, male Long-Evans rats were pretreated with AM251 (0, 10, 100, or 200 µg, i.c.v.), followed by CRF (0 or 0.5 µg, i.c.v.), before testing for anxiety-like behavior in the elevated plus maze (EPM). In Experiment 2, rats were exposed to cocaine (20 mg/kg, i.p.) or saline for 14 consecutive days. Forty-eight hours following cocaine exposure, rats were pretreated with AM251 (0, 10, or 100 µg, i.c.v.) and tested in the EPM. AM251 produced an anxiogenic response at the highest dose, but reversed the behavioral anxiety induced by CRF and withdrawal from chronic cocaine in a dose-dependent manner. AM251 also increased plasma corticosterone levels, but did so irrespective of CRF treatment or cocaine preexposure. Our findings suggest that the anxiogenic effects of CRF and cocaine withdrawal are mediated, at least in part, by CB1 receptor transmission, and provide evidence in support of eCB-CRF interactions that are independent of the hypothalamic-pituitary-adrenal axis.
Assuntos
Ansiedade/tratamento farmacológico , Cocaína/efeitos adversos , Hormônio Liberador da Corticotropina/farmacologia , Inibidores da Captação de Dopamina/efeitos adversos , Receptor CB1 de Canabinoide/antagonistas & inibidores , Síndrome de Abstinência a Substâncias/tratamento farmacológico , Animais , Ansiedade/sangue , Ansiedade/induzido quimicamente , Comportamento Animal/efeitos dos fármacos , Corticosterona/sangue , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Masculino , Piperidinas/farmacologia , Sistema Hipófise-Suprarrenal/efeitos dos fármacos , Pirazóis/farmacologia , Ratos , Ratos Long-EvansRESUMO
Wernicke encephalopathy is a neurological disorder, often caused by a lack of thiamine. Immediate diagnosis and therapy are important to prevent significant morbidity and mortality. Based on a clinical case radiologic imaging features on MRI are demonstrated.
Assuntos
Encefalopatia de Wernicke , Idoso , Alcoolismo/complicações , Humanos , Imageamento por Ressonância Magnética/métodos , Masculino , Desnutrição/complicações , Tiamina/administração & dosagem , Tiamina/uso terapêutico , Encefalopatia de Wernicke/diagnóstico , Encefalopatia de Wernicke/tratamento farmacológico , Encefalopatia de Wernicke/etiologiaRESUMO
The androgen receptor (AR) plays a crucial role in the modulation of prostate cell proliferation and is involved in the development and progression of prostate cancer (PCa). An understanding of the complex regulation of AR provides novel treatment options for PCa. Here, we show (i) that the ubiquitin-like modifier, interferon-stimulated gene 15 (ISG15), and most enzymes involved in ISG15 conjugation were upregulated in tumor samples versus in non-malignant tissues of PCa patients and (ii) that the expression of these components significantly differed between tumors in patients treated with and without androgen ablation. Using PCa cell lines as in vitro models, the specific androgen-mediated, AR-dependent regulation of the ISGylation components was confirmed. In addition, the ISGylation system controls AR mRNA and protein expressions, as overexpression of Ube1L as a limiting ISGylation factor in the AR(+) androgen-sensitive PCa cell line, LNCaP, results in significant AR upregulation, accompanied by an increased proliferation even under androgen deprivation. Accordingly, Ube1L knockdown decreased the AR expression. Thus, this study describes for the first time the modulation of AR expression by ISGylation components, which affects the proliferation of PCa cells, thereby providing evidence for a novel function of the ISGylation system in malignant transformation.
Assuntos
Citocinas/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias da Próstata/metabolismo , Ubiquitinas/genética , Androgênios/fisiologia , Citocinas/fisiologia , Técnicas de Silenciamento de Genes , Humanos , Masculino , Neoplasias da Próstata/patologia , RNA Mensageiro/genética , Receptores Androgênicos/genética , Receptores Androgênicos/metabolismo , Receptores Androgênicos/fisiologia , Transdução de Sinais , Ubiquitinas/fisiologiaRESUMO
We report on a twenty-three year old woman with shifting polyarthritis after an episode with gastroenteritis. Despite normalisation of stool we could detect campylobacter jejuni one month later. After therapy with macrolide antibiotics arthritis recovered.
Assuntos
Artrite Reativa/diagnóstico , Infecções por Campylobacter/complicações , Campylobacter jejuni , Diarreia/complicações , Adulto , Diagnóstico Diferencial , Feminino , HumanosRESUMO
We report a case of a 49-year-old woman from the Philippines, who has been living for many years in Switzerland, with chronic unilateral swelling of the cervical lymphnodes. After the serological exclusion of several microorganisms, a fine needle aspiration was performed without conclusive result. However, patient history and a strongly positive PPD skin test were highly suspicious for tuberculous lymphadenitis, which was finally confirmed by lymphnode biopsy on histopathological and cultural workup. Therefore tuberculostatic treatment was initiated for 6 months.
Assuntos
Emigrantes e Imigrantes , Doenças Linfáticas/etiologia , Tuberculose dos Linfonodos/diagnóstico , Algoritmos , Diagnóstico Diferencial , Feminino , Humanos , Excisão de Linfonodo , Linfonodos/patologia , Doenças Linfáticas/patologia , Pessoa de Meia-Idade , Filipinas/etnologia , Suíça , Tuberculose dos Linfonodos/patologiaRESUMO
Malakoplakia is a rare inflammatory condition seen in transplant patients. There are two previously reported cases of malakoplakia involving the gastrointestinal tract in liver transplant patients. The present paper reports a case of colonic malakoplakia in a 58-year-old woman, a liver transplant recipient who was receiving immunosuppressive drugs. She presented with chronic diarrhea while on tacrolimus. There was no history of antecedent infection. Colonoscopy showed patchy mucosal edema, but no discrete yellow plaques or nodules. The diagnosis was made by colon biopsies, which showed chronic inflammation with many histiocytes containing Michaelis-Gutmann bodies. Although rare, malakoplakia is one of many potential causes of diarrhea in a transplant patient. The present case indicates that malakoplakia may be associated with chronic diarrhea, even if there are no macroscopic lesions seen during colonoscopy.