RESUMO
OBJECTIVES: To assess effectiveness and safety of damoctocog alfa pegol in interim analyses of the ongoing real-world hemophilia A HEM-POWR study. METHODS: HEM-POWR (NCT03932201) is a multinational Phase 4 prospective observational study. The primary objective was annualized bleeding rate (ABR) in previously treated patients (PTPs) with hemophilia A. Secondary objectives included adverse events and number of affected joints. RESULTS: At data cut-off (August 17, 2022), the safety analysis set included 268 patients and the full analysis set (FAS) included 161 patients. The most common dosing regimen during observation period was prophylaxis (FAS = 158/161, 98.1%) every 3-4 days (twice weekly; FAS = 78/158, 49.4%) and a median (min, max) infusion dose of 37.5 (10, 72) IU/kg. PTPs receiving prophylactic damoctocog alfa pegol have fewer infusions compared with prior treatment. Median total ABR (Q1, Q3) was 0.0 (0.0, 1.8) and mean total ABR (SD) was 2.4 (8.2). The proportion of patients with no affected joints increased between initial visit and follow-up. No FVIII inhibitors, treatment-related adverse events, or deaths were reported. CONCLUSIONS: Damoctocog alfa pegol shows effectiveness and acceptable safety, as well as consistent utilization, in real-world PTPs with hemophilia A, including in patients with non-severe hemophilia and those with a history of inhibitors. Please see video for a summary of this study.
Assuntos
Hemofilia A , Humanos , Hemofilia A/complicações , Hemofilia A/tratamento farmacológico , Estudos Prospectivos , Fator VIII/efeitos adversos , Esquema de MedicaçãoRESUMO
Bacteria contaminate semen during collection and handling. The objective of this study was to identify the bacteria in pony stallion semen, the effects of antibiotics included in commercial semen extenders (lincomycin and spectinomycin) and the effect of modified single layer centrifugation (MSLC), on bacterial load. Ejaculates from six pony stallions, 3 ejaculates per animal, were extended in EquiPlus extender either with or without antibiotics. Aliquots were processed by MSLC to form four treatment groups: control and MSLC with antibiotics (CA and SA, respectively) and control and MSLC without antibiotics (CW and SW, respectively). Bacteriological examinations were carried out within 2 hr. Thirty-one species of bacteria were isolated from one or more ejaculates, with Corynebacterium spp. being the most frequently detected. Corynebacterium spp. were present in all ejaculates. The MSLC resulted in a significantly lower total bacterial count than controls (CA vs. SA, p < 0.001; CW vs. SW, p < 0.0001).
Assuntos
Antibacterianos/farmacologia , Bactérias/isolamento & purificação , Centrifugação/métodos , Sêmen/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Animais , Bactérias/efeitos dos fármacos , Carga Bacteriana , Ejaculação , Cavalos , Masculino , Sêmen/microbiologia , Preservação do Sêmen/métodos , Preservação do Sêmen/veterinária , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/microbiologiaRESUMO
A robust liquid chromatography-tandem mass spectrometry method was developed and comprehensively validated for the quantification of cefquinome considering the changing matrix composition from bovine colostrum to raw milk. Sample preparation consisted of addition of isotopically labeled cefquinome internal standard prior to protein precipitation of 2 g colostrum or milk followed by solid-phase extraction. A wide concentration range from 1 to 5000 ng cefquinome per gram of colostrum or milk was quantified using a 3200 QTRAP tandem mass spectrometer in positive ionization mode with electrospray ionization. Validation was performed according to the European Commission Decision 2002/657/EC guidelines. Matrix-comprehensive in-house validation included analytical limits CCα and CCß, recovery, precision and calibration curves with prediction intervals, storage conditions, and evaluation of robustness based on factorial effect analysis. The detection limit was 0.2 ng cefquinome per gram of colostrum or milk. Recovery was between 98.4 and 99.4% for cefquinome concentrations from 4 to 240 ng/g. None of the investigated validation factors (matrix, storage of extracts, lot of SPE cartridges, and operators) exerted an influence higher than ± 3.2%, indicating that these factors make relatively low contributions to the respective combined measurement uncertainties. The comprehensively validated method enables routine residue control purposes and to monitor the pharmacokinetics of cefquinome in bovine colostrum and raw milk. In particular, residue depletion curves of cefquinome from high concentrations in first milking after treatment to concentrations far below the maximum residue limit can be measured.
Assuntos
Bovinos/fisiologia , Cefalosporinas/química , Cromatografia Líquida/métodos , Colostro/química , Leite/química , Espectrometria de Massas em Tandem/métodos , Animais , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Manejo de Espécimes , Fatores de TempoRESUMO
Streptomyces violaceoruber grown in co-culture with Streptomyces aburaviensis produces an about 17-fold higher volume of droplets on its aerial mycelium than in single-culture. Physical separation of the Streptomyces strains by either a plastic barrier or by a dialysis membrane, which allowed communication only by the exchange of volatile compounds or diffusible compounds in the medium, respectively, still resulted in enhanced droplet formation. The application of molecular sieves to bioassays resulted in the attenuation of the droplet-inducing effect of S. aburaviensis indicating the absorption of the compound. 1H-NMR analysis of molecular-sieve extracts and the selective indophenol-blue reaction revealed that the volatile droplet-inducing compound is ammonia. The external supply of ammonia in biologically relevant concentrations of ≥8 mM enhanced droplet formation in S. violaceoruber in a similar way to S. aburaviensis. Ammonia appears to trigger droplet production in many Streptomyces strains because four out of six Streptomyces strains exposed to ammonia exhibited induced droplet production.
Assuntos
Amônia/metabolismo , Gotículas Lipídicas/química , Streptomyces/química , Amônia/análise , Amônia/química , Bioensaio , Difusão , Gases/química , Concentração de Íons de Hidrogênio , Espectroscopia de Ressonância Magnética , Compostos Orgânicos/química , Streptomyces/crescimento & desenvolvimento , Streptomyces/metabolismoRESUMO
BACKGROUND/AIMS: Human peritoneal mesothelial cells (HPMC) secrete phosphatidylcholines (PC) which form a lipid bilayer lining the peritoneum. They prevent frictions and adhesions and act as a barrier to the transport of water-soluble solutes while permitting water flux. PC may play an essential role in peritoneal integrity and function, the role of PD induced HPMC senescence on PC homeostasis, however, is unknown. METHODS: HPMC cell lines were isolated from four non-uremic patients. Expression of the three PC synthesis genes (rt-PCR), and cellular storage and secretion of PC (ESI-mass-spectrometry) were analyzed in young and senescent HPMC (>Hayflick-limit). RESULTS: Senescent cells displayed significantly altered morphology; flow cytometry demonstrated extensive staining for senescence-associated beta galactosidase. Nine different PC were detected in HPMC with palmitoyl-myristoyl phosphatidylcholine (PMPC) being most abundant. In senescent HPMC mRNA expression of the three key PC synthesis genes was 1.5-, 2.4- and 6-fold increased as compared to young HPMC, with the latter, phosphatidylcholine cytidylyltransferase, being rate limiting. Intracellular storage of the nine PC was 75-450 % higher in senescent vs. young HPMC, PC secretion rates were 100-300 % higher. Intracellular PC concentrations were not correlated with the PC secretion rates. Electron microscopy demonstrated lamellar bodies, the primary storage site of PC, in senescent but not in young cells. CONCLUSION: Senescent HPMC store and secrete substantially more PC than young cells. Our findings indicate a novel protective mechanism, which should counteract peritoneal damage induced by chronic exposure to PD fluids.
Assuntos
Senescência Celular , Células Epiteliais/metabolismo , Epitélio/metabolismo , Peritônio/metabolismo , Fosfatidilcolinas/metabolismo , Linhagem Celular , Células Epiteliais/ultraestrutura , Humanos , Peritônio/citologiaRESUMO
Steroid-refractory graft-versus-host disease is a life-threatening complication after allogeneic stem cell transplantation. Evidence is accumulating that steroid-refractory graft-versus-host disease is associated with endothelial distress. Endothelial cell homeostasis is regulated by nitric oxide, and serum nitrates are derived from nitric oxide synthase activity or dietary sources. In this retrospective study based on 417 patients allografted at our institution we investigated whether quantification of serum nitrates could predict steroid-refractory graft-versus-host disease. Elevated pre-transplant levels of serum nitrates (>26.5 µM) predicted steroid-refractory graft-versus-host disease (P=0.026) and non-relapse mortality (P=0.028), particularly in combination with high pre-transplant angiopoietin-2 levels (P=0.0007 and P=0.021, respectively). Multivariate analyses confirmed serum nitrates as independent predictors of steroid-refractory graft-versus-host disease and non-relapse mortality. Differences in serum nitrate levels did not correlate with serum levels of tumor necrosis factor or C-reactive protein or expression of inducible nitric oxide synthase in blood cells. Patients with high pre-transplant nitrate levels had significantly reduced rates of refractory graft-versus-host disease (P=0.031) when pravastatin was taken. In summary, patients at high risk of developing steroid-refractory graft-versus-host disease could be identified prior to transplantation by serum markers linked to endothelial cell function. Retrospectively, statin medication was associated with a reduced incidence of refractory graft-versus-host disease in this endothelial high-risk cohort.
Assuntos
Doença Enxerto-Hospedeiro/diagnóstico , Doença Enxerto-Hospedeiro/etiologia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Nitratos/sangue , Adolescente , Adulto , Idoso , Angiopoietina-2/sangue , Ativação Enzimática , Feminino , Doença Enxerto-Hospedeiro/tratamento farmacológico , Doença Enxerto-Hospedeiro/prevenção & controle , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Masculino , Pessoa de Meia-Idade , Óxido Nítrico Sintase/metabolismo , Avaliação de Resultados da Assistência ao Paciente , Prognóstico , Recidiva , Estudos Retrospectivos , Risco , Esteroides/uso terapêutico , Adulto JovemRESUMO
A sensitive and robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method allowing the rapid screening and confirmation of thyreostatic drugs in bovine blood plasma was developed and validated according to Commission Decision 2002/657/EC, chapter 3.1.3 "alternative validation", by applying a matrix-comprehensive in-house validation concept. Decision limit CCα, detection capability CCß, recovery, repeatability, within-laboratory reproducibility and the uncertainty of measurement were calculated. Furthermore, a factorial effect analysis was carried out to identify factors that have a significant influence on the method. Factors considered to be relevant for the method in routine analysis (e.g. operator, storage duration of the extracts before measurement, different cartridge lots and duration of sample preparation) were systematically varied on two levels during the validation study. Subsequently, the extent to which these factors influence the measurement results of the individual analytes was examined.
Assuntos
Antitireóideos/sangue , Análise Química do Sangue/métodos , Cromatografia Líquida/normas , Espectrometria de Massas em Tandem/normas , Animais , Bovinos , Limite de Detecção , Estrutura Molecular , Reprodutibilidade dos TestesRESUMO
A sensitive and robust liquid chromatography-tandem mass spectrometry method allowing the rapid screening and confirmation of ten synthetic corticosteroids in bovine and porcine muscle tissue was developed and validated. The validation was conducted according to Commission Decision 2002/657/EC, Sect. 3.1.3 ("Validation according to alternative models"), by applying a matrix-comprehensive in-house validation concept. The decision limit, detection capability, recovery, repeatability, within-laboratory-reproducibility and measurement uncertainty were calculated. Furthermore, a factorial effect analysis was conducted to identify factors that have a significant influence on the method. To this end, factors considered to be relevant for the method in routine analysis (e.g. operator, duration of storage of the extracts before measurement, different lots of the cartridges and different species) were systematically varied on two levels during the validation study. Subsequently, the extent to which these factors influence the measurement results of the individual analytes was examined.
Assuntos
Corticosteroides/análise , Cromatografia Líquida de Alta Pressão/métodos , Resíduos de Drogas/análise , Contaminação de Alimentos/análise , Carne/análise , Músculo Esquelético/química , Espectrometria de Massas em Tandem/métodos , Animais , Bovinos , SuínosRESUMO
Degradation of the lower chlorinated ethenes is crucial to the application of natural attenuation or in situ bioremediation on chlorinated ethene contaminated sites. Recently, within mixtures of several chloroethenes as they can occur in contaminated groundwater inhibiting effects on aerobic chloroethene degradation have been shown. The current study demonstrated that metabolic vinyl chloride (VC) degradation by an enrichment culture originating from groundwater was not affected by an equimolar concentration (50 µM) of cis-1,2-dichloroethene (cDCE). Only cDCE concentrations at a ratio of 2.4:1 (initial cDCE to VC concentration) caused minor inhibition of VC degradation. Furthermore, the degradation of VC was not affected by the presence of trans-1,2-dichloroethene (tDCE), 1,1-dichloroethene (1,1-DCE), trichloroethene (TCE), and tetrachloroethene (PCE) in equimolar concentrations (50 µM). Only cDCE and tDCE were cometabolically degraded in small amounts. The VC-degrading culture demonstrated a broad pH tolerance from 5 to 9 with an optimum between 6 and 7. Results also showed that the culture could degrade VC concentrations up to 1,800 µM (110 mg/L).
Assuntos
Bactérias/citologia , Bactérias/metabolismo , Técnicas de Cultura de Células/métodos , Cloreto de Vinil/metabolismo , Aerobiose , Biodegradação Ambiental , Dicloroetilenos/metabolismo , Concentração de Íons de HidrogênioRESUMO
Both obesity and sarcopenia are frequently associated in ageing, and together may promote the progression of related conditions such as diabetes and frailty. However, little is known about the pathophysiological mechanisms underpinning this association. Here we show that systemic alanine metabolism is linked to glycaemic control. We find that expression of alanine aminotransferases is increased in the liver in mice with obesity and diabetes, as well as in humans with type 2 diabetes. Hepatocyte-selective silencing of both alanine aminotransferase enzymes in mice with obesity and diabetes retards hyperglycaemia and reverses skeletal muscle atrophy through restoration of skeletal muscle protein synthesis. Mechanistically, liver alanine catabolism driven by chronic glucocorticoid and glucagon signalling promotes hyperglycaemia and skeletal muscle wasting. We further provide evidence for amino acid-induced metabolic cross-talk between the liver and skeletal muscle in ex vivo experiments. Taken together, we reveal a metabolic inter-tissue cross-talk that links skeletal muscle atrophy and hyperglycaemia in type 2 diabetes.
Assuntos
Alanina/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Hiperglicemia/metabolismo , Fígado/metabolismo , Músculo Esquelético/patologia , Atrofia Muscular/metabolismo , Alanina/sangue , Alanina Transaminase/sangue , Animais , Glicemia/metabolismo , Diabetes Mellitus Tipo 2/patologia , Modelos Animais de Doenças , Homeostase , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Obesidade/metabolismoRESUMO
BACKGROUND: Residual/recurrent mitral valve regurgitation is observed in 30% after undersized ring annuloplasty (RING) for ischemic mitral regurgitation (IMR). RING addresses primarily annular dilatation but does not correct severe leaflet tethering attributable to papillary muscle (PM) displacement. We proposed adjunctive PM repositioning under transesophageal echocardiography (TEE) guidance in the loaded beating heart using a transventricular suture (STRING). METHODS AND RESULTS: Patients with tenting height > or =10 mm were identified as high-risk patients for repair failure. In these patients (n=30, age 68+/-11 years, ejection fraction 37+/-14%), RING (partial, median 29 mm) was combined with the adjunctive STRING-technique. A Teflon-pledgeted 3-0-polytetrafluoroethylene-suture was anchored in the posterior PM via horizontal aortotomy, exteriorized through the aorto-mitral continuity, and tied in the loaded beating heart under TEE guidance. Tenting height (14+/-2 mm versus 6+/-1 mm, P<0.001) and tenting area (3.9+/-0.9 cm(2) versus 1.0+/-0.2 cm(2), P<0.001) decreased. The distance between pPM and aorto-mitral continuity decreased (44+/-4 mm versus 37+/-3 mm, P<0.001). Survival at 2 years was similar compared with a historical matched control-group (89% versus 73%, P=0.13), whereas freedom from MR>II was higher in the RING+STRING-group (94% versus 71%, P=0.01). End-diastolic (61.7+/-7.2 mm versus 54.8+/-9.2 mm, P<0.001) and end-systolic (48.5+/-8.5 mm versus 42.7+/-7.8 mm, P=0.002) ventricular diameters decreased in the RING+STRING-group but persisted in the control-group (60.4+/-7.8 mm versus 58.9+/-7.5 mm, P=0.38; 47.8+/-9.6 mm versus 48.3+/-9.5 mm, P=0.52). During follow-up (median 26 months) only 1 patient of the study-group required reoperation for degenerative MR, while 2 control-group patients underwent reoperation for recurrent functional MR. CONCLUSIONS: Our novel approach for IMR attenuates high risk of repair failure in patients with severe leaflet tethering and results in reverse remodeling.
Assuntos
Insuficiência da Valva Mitral/cirurgia , Valva Mitral/cirurgia , Isquemia Miocárdica/complicações , Técnicas de Sutura , Idoso , Ecocardiografia , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Insuficiência da Valva Mitral/diagnóstico por imagem , Isquemia Miocárdica/diagnóstico por imagemRESUMO
OBJECTIVE: Recent studies have shown an increase of C-reactive-protein (CRP) after exposure to zinc- and copper-containing welding fumes. The objective of this study was to determine the effects of exposure to zinc- and copper-containing welding fumes on leukocytes, their subtypes, and myeloperoxidase (MPO). METHODS: Serum samples of male volunteers were examined after exposures to welding fumes in two settings: repeated exposure on 4 consecutive days for 6âhours and single exposures for different times (3, 4, 5âhours). RESULTS: Neutrophil granulocyte and MPO levels showed increases 24âhours after single and repeated exposures for 6âhours similar to CRP increases reported in literature. Overall leukocyte levels and levels of monocytes and lymphocytes were not significantly affected. CONCLUSIONS: This study indicates the involvement of neutrophil granulocytes in welding fume fever additional to mediator related effects.
Assuntos
Poluentes Ocupacionais do Ar , Cobre/efeitos adversos , Neutrófilos/citologia , Exposição Ocupacional , Soldagem , Zinco/efeitos adversos , Poluentes Ocupacionais do Ar/efeitos adversos , Biomarcadores , Humanos , Inflamação , Exposição por Inalação/efeitos adversos , Masculino , Neutrófilos/enzimologia , Exposição Ocupacional/efeitos adversos , Peroxidase/análiseRESUMO
Chloroethenes belong to the most widely distributed groundwater contaminants. Since 2014, it has been known that trichloroethene (TCE) can be degraded aerobically and metabolically as growth substrate by a mixed bacterial enrichment culture (named SF culture). In this study, the degradation capabilities under a range of field-relevant conditions were investigated in fixed-bed reactors as well as in batch experiments. Aerobic metabolic TCE degradation was stable over the long term, with degradation optima at 22⯰C and pH 7. Degradation of up to 400⯵M TCE was observed. The longest starvation period after which degradation of TCE was regained was 112 days. The possible co-contaminants perchloroethene, trans-1,2-dichloroethene, and cis-1,2-dichloroethene did not inhibit TCE degradation, even though they were not degraded themselves. The presence of equimolar amounts of 1,1-dichloroethene and vinyl chloride inhibited TCE degradation. Experiments with groundwater from different chloroethene-contaminated field sites proved the potential of the SF culture for bioaugmentation. Thus, aerobic metabolic TCE degradation should be considered as a promising method for the bioremediation of field sites with TCE as the main contaminant.
Assuntos
Água Subterrânea , Tricloroetileno , Cloreto de Vinil , Biodegradação AmbientalRESUMO
Stilbenes and zeranol are nonsteroidal estrogenic growth promoters which are banned in the European Union (EU) for use in food-producing animals by Council Directive 96/22/EC. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the screening and confirmation of stilbenes (diethylstilbestrol, dienestrol, hexestrol) and resorcylic acid lactones (zeranol and its metabolites taleranol and zearalanone as well as the mycotoxins alpha-zearalenol, beta-zearalenol and zearalenone) in bovine urine. The method permits the confirmation and quantification of stilbenes and resorcylic acid lactones at levels below 1 microg L(-1) and 1.5 microg L(-1), respectively. The validation was carried out according to Commission Decision 2002/657/EC, Chap. 3.1.3 "alternative validation" by a matrix-comprehensive in-house validation concept. Decision limit CCalpha, detection capability CCbeta, recovery, repeatability, within-laboratory reproducibility and the uncertainty of measurement were calculated. Furthermore, a factorial effect analysis was carried out to identify factors that have a significant influence on the method. Factors considered to be relevant for the method in routine analysis (e.g. operator, matrix condition, storage duration of the extracts before measurement, different cartridge lots, hydrolysis conditions) were systematically varied on two levels. The factorial analysis showed that different cartridge lots, storage durations and matrix conditions can exert a relevant influence on the method.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Cromatografia por Troca Iônica/métodos , Hidroxibenzoatos/química , Lactonas/urina , Espectrometria de Massas por Ionização por Electrospray/métodos , Estilbenos/urina , Espectrometria de Massas em Tandem/métodos , Animais , Bovinos , Feminino , Lactonas/química , Masculino , Estrutura Molecular , Reprodutibilidade dos Testes , Estilbenos/químicaRESUMO
Assessing changes in the isotopic signature of contaminants is a promising new tool to monitor microbial degradation processes. In this study, chloroethene degradation was proven by depletion of chloroethenes, formation of chloride, increase in protein content and stable carbon isotope fractionation. Aerobic degradation of vinyl chloride (VC) was found to proceed metabolically, with degradation rates of 0.48 and 0.29 d(-1); and growth yields of 9.7 and 6.4 g of protein/mol of VC at room and groundwater temperature, respectively. Cis-1,2-dichloroethene (cDCE) was degraded cometabolically under aerobic conditions when VC was provided as growth substrate. Aerobic degradation was associated with significant stable carbon isotope fractionation, with enrichment factors ranging from -5.4+/-0.4 per thousand for metabolic degradation of VC to -9.8+/-1.7 per thousand for cometabolic degradation of cDCE. Thus, it was demonstrated that stable carbon isotope fractionation is suitable for assessing aerobic chloroethene degradation, which can contribute significantly to site remediation.
Assuntos
Bactérias/crescimento & desenvolvimento , Fracionamento Químico , Dicloroetilenos/metabolismo , Cloreto de Vinil/metabolismo , Aerobiose , Anaerobiose , Biodegradação Ambiental , Isótopos de Carbono , CinéticaRESUMO
A sensitive and robust LC-MS/MS method allowing the rapid screening and confirmation of selective androgen receptor modulators in bovine urine was developed and successfully validated according to Commission Decision 2002/657/EC, chapter 3.1.3 'alternative validation', by applying a matrix-comprehensive in-house validation concept. The confirmation of the analytes in the validation samples was achieved both on the basis of the MRM ion ratios as laid down in Commission Decision 2002/657/EC and by comparison of their enhanced product ion (EPI) spectra with a reference mass spectral library by making use of the QTRAP technology. Here, in addition to the MRM survey scan, EPI spectra were generated in a data-dependent way according to an information-dependent acquisition criterion. Moreover, stability studies of the analytes in solution and in matrix according to an isochronous approach proved the stability of the analytes in solution and in matrix for at least the duration of the validation study. To identify factors that have a significant influence on the test method in routine analysis, a factorial effect analysis was performed. To this end, factors considered to be relevant for the method in routine analysis (e.g. operator, storage duration of the extracts before measurement, different cartridge lots and different hydrolysis conditions) were systematically varied on two levels. The examination of the extent to which these factors influence the measurement results of the individual analytes showed that none of the validation factors exerts a significant influence on the measurement results.
Assuntos
Moduladores Seletivos de Receptor Estrogênico/urina , Espectrometria de Massas em Tandem/normas , Animais , Bovinos , Cromatografia Líquida/normasRESUMO
The hypothesis in this study was continuous treatment of stallions with the GnRH agonist deslorelin inhibits reproductive functions. A 2-week pre-experimental period was followed by an 11-week deslorelin implant treatment. Stallions received 4.7 (D1, n = 7), or 18.8â¯mg deslorelin (D2, n = 5) or remained untreated (C, n = 5). Libido, sperm motility, membrane integrity, DNA fragmentation, estrogen receptors, basal plasma testosterone and Anti Muellerian hormone (AMH) concentrations were evaluated once weekly during the treatment period. The testosterone response to the GnRH agonist buserelin and hCG was evaluated twice. In Week 2, stallions in Group C but not Groups D1 and D2 responded to buserelin with testosterone release (P < 0.001), while in Week 9, stallions in Group C and D1 but not D2 released testosterone after buserelin administration (group P < 0.01, week Pâ¯=â¯0.01). Stallions of all groups responded to hCG with testosterone release at both times of hCG administration (P < 0.001). The AMH concentration was similar in all groups. Deslorelin thus reduced pituitary responsiveness to GnRH but only with a large dose and this effect persisted for several weeks. Total sperm count increased transiently with the D2 treatment but not in stallions of the D1 and C groups after implant insertion (time P < 0.01, time x group P < 0.001). The percentage of ESR1-positive spermatozoa decreased transiently in Group D2 (time P < 0.01, time × group P < 0.01). There was no difference among groups at any time during the study in percentage of motile and membrane-intact spermatozoa and sperm with DNA fragmentation. In conclusion, deslorelin implants modulate pituitary function in stallions but not to an extent that affects testicular function.
Assuntos
Implantes de Medicamento/farmacologia , Cavalos , Testosterona/sangue , Pamoato de Triptorrelina/análogos & derivados , Animais , Implantes de Medicamento/administração & dosagem , Inibidores Enzimáticos/administração & dosagem , Inibidores Enzimáticos/farmacologia , Masculino , Sêmen , Análise do Sêmen/veterinária , Espermatogênese/efeitos dos fármacos , Testosterona/metabolismo , Pamoato de Triptorrelina/administração & dosagem , Pamoato de Triptorrelina/farmacologiaRESUMO
Contamination of semen with bacteria arises during semen collection and handling. This bacterial contamination is typically controlled by adding antibiotics to semen extenders but intensive usage of antibiotics can lead to the development of bacterial resistance and may be detrimental to sperm quality. The objective of this study was to determine the effects of antibiotics in a semen extender on sperm quality and to investigate the effects of removal of bacteria by modified Single Layer Centrifugation (MSLC) through a colloid. Semen was collected from six adult pony stallions (three ejaculates per male). Aliquots of extended semen were used for MSLC with Equicoll, resulting in four treatment groups: control and MSLC in extender with antibiotics (CA and SA, respectively); control and MSLC in extender without antibiotics (CW and SW, respectively). Sperm motility, membrane integrity, mitochondrial membrane potential and chromatin integrity were evaluated daily by computer-assisted sperm analysis (CASA) and flow cytometry. There were no differences in sperm quality between CA and CW, or between SA and SW, although progressive motility was negatively correlated to total bacterial counts at 0 h. However, MSLC groups showed higher mean total motility (P < 0.001), progressive motility (P < 0.05), membrane integrity (P < 0.0001) and mitochondrial membrane potential (P < 0.05), as well as better chromatin integrity (P < 0.05), than controls. Sperm quality remained higher in the MSLC groups than controls throughout storage. These results indicate that sperm quality was not adversely affected by the presence of antibiotics but was improved considerably by MSLC.
RESUMO
We hypothesized that housing of stallions in a thermoneutral temperature zone during autumn and winter does not only influence metabolism and hair shedding but also improves the characteristics of raw and processed semen. Fertile Shetland pony stallions were followed from October to June. This time coincided with the seasons autumn, winter and spring. Ponies were kept in outside paddocks (group CON, n = 8) or in indoor stables (group ST, n = 8) from October to March when ST stallions returned to outdoor paddocks, but ponies remained in the same groups. The rectal temperature was measured once weekly. Heart rate, heart rate variability, testosterone and cortisol concentration in blood as well as quality and length of the coat were determined. Semen was collected once weekly and raw semen characteristics were analyzed. The characteristics of cooled-stored and cryopreserved semen were determined once monthly. During the stabling period, environmental temperature for group ST averaged 13.6 ± 2.3 and for group CON 5.6 ± 4.2 °C. The mean rectal temperature was higher (p < 0.05) in ST than in CON stallions. All hair coat parameters underwent seasonal changes (p < 0.001) and differed between groups (p < 0.05) with shorter guard hair, slower hair regrowth and earlier hair change in ST stallions. Season influenced heart rate which was highest in autumn, lowest in winter and intermediate in spring but did not differ between groups. Testosterone and cortisol concentrations in blood as well as sexual behavior underwent seasonal changes but did not differ between CON and ST stallions. Gel-free semen volume and total sperm count were influenced by season (p < 0.01) and showed a more pronounced increase from winter to spring in CON than in ST stallions (p < 0.05) while no differences with regard to sperm concentration in raw semen were detected. Progressive motility of spermatozoa in raw semen was highest in spring (p < 0.05) but not affected by group. In cooled-stored and cryopreserved semen, neither season nor group affected total motility, progressive motility or membrane integrity. In conclusion, environmental temperature during autumn and winter had clear results on body temperature as well as hair coat characteristics in Shetland stallions. Simultaneously determined effects on semen characteristics were minimal indicating that reproductive function in the horse is more dependent on day length i.e. the geophysical year than on other environmental factors.
Assuntos
Cabelo/fisiologia , Cavalos/fisiologia , Hidrocortisona/sangue , Estações do Ano , Temperatura , Testosterona/sangue , Animais , Temperatura Corporal , Estudos de Casos e Controles , Abrigo para Animais , Masculino , Análise do Sêmen/veterinária , Comportamento Sexual Animal/fisiologiaRESUMO
Elevated serum trimethylamine-N-oxide (TMAO) was previously reported to be associated with an elevated risk for cardiovascular events. TMAO originates from the microbiota-dependent breakdown of food-derived phosphatidylcholine (PC) to trimethylamine (TMA), which is oxidized by hepatic flavin-containing monooxygenases to TMAO. Our aim was to investigate the predominant site of absorption of the bacterial PC-breakdown product TMA. A healthy human proband was exposed to 6.9 g native phosphatidylcholine, either without concomitant treatment or during application with the topical antibiotic rifaximin, or exposed only to 6.9 g of a delayed-release PC formulation. Plasma and urine concentrations of TMA and TMAO were determined by electrospray ionization tandem mass spectrometry (plasma) and gas chromatography-mass spectrometry (urine). Native PC administration without concomitant treatment resulted in peak plasma TMAO levels of 43 ± 8 µM at 12 h post-ingestion, which was reduced by concomitant rifaximin treatment to 22 ± 8 µM (p < 0.05). TMAO levels observed after delayed-release PC administration were 20 ± 3 µM (p < 0.001). Accordingly, the peak urinary concentration at 24 h post-exposure dropped from 252 ± 33 to 185 ± 31 mmol/mmol creatinine after rifaximin treatment. In contrast, delayed-release PC resulted in even more suppressed urinary TMAO levels after the initial 12-h observation period (143 ± 18 mmol/mmol creatinine) and thereafter remained within the control range (24 h: 97 ± 9 mmol/mmol creatinine, p < 0.001 24 h vs. 12 h), indicating a lack of substrate absorption in distal intestine and large bowel. Our results showed that the microbiota in the small intestine generated the PC breakdown product TMA. The resulting TMAO, as a cardiovascular risk factor, was suppressed by topical-acting antibiotics or when PC was presented in an intestinally delayed release preparation.