RESUMO
OBJECTIVES: Although controversy exists regarding the efficacy of antibiotic prophylaxis for patients at risk of infective endocarditis, expert committees continue to publish recommendations for antibiotic prophylaxis regimens. This study aimed to evaluate the efficacy of four antimicrobial regimens for the prevention of bacteraemia following dental extractions. METHODS: The study population included 266 adults requiring dental extractions who were randomly assigned to the following five groups: control (no prophylaxis); 1000/200 mg of amoxicillin/clavulanate intravenously; 2 g of amoxicillin by mouth; 600 mg of clindamycin by mouth; and 600 mg of azithromycin by mouth. Venous blood samples were collected from each patient at baseline and at 30 s, 15 min and 1 h after dental extractions. Samples were inoculated into BACTEC Plus culture bottles and processed in the BACTEC 9240. Conventional microbiological techniques were used for subcultures and further identification of the isolated bacteria. The trial was registered at ClinicalTrials.gov with ID number NCT02115776. RESULTS: The incidence of bacteraemia in the control, amoxicillin/clavulanate, amoxicillin, clindamycin and azithromycin groups was: 96%, 0%, 50%, 87% and 81%, respectively, at 30 s; 65%, 0%, 10%, 65% and 49% at 15 min; and 18%, 0%, 4%, 19% and 18% at 1 h. Streptococci were the most frequently identified bacteria. The percentage of positive blood cultures at 30 s post-extraction was lower in the amoxicillin/clavulanate group than in the amoxicillin group (Pâ<â0.001). The incidence of bacteraemia in the clindamycin group was similar to that in the control group. CONCLUSIONS: Bacteraemia following dental extractions was undetectable with amoxicillin/clavulanate prophylaxis. Alternative antimicrobial regimens should be sought for patients allergic to the ß-lactams.
Assuntos
Combinação Amoxicilina e Clavulanato de Potássio/administração & dosagem , Antibacterianos/administração & dosagem , Antibioticoprofilaxia/métodos , Bacteriemia/prevenção & controle , Extração Dentária/efeitos adversos , Inibidores de beta-Lactamases/administração & dosagem , Administração Intravenosa , Adolescente , Adulto , Técnicas Bacteriológicas , Sangue/microbiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Resultado do Tratamento , Adulto JovemRESUMO
Neither breakpoints (BPs) nor epidemiological cutoff values (ECVs) have been established for Candida spp. with anidulafungin, caspofungin, and micafungin when using the Sensititre YeastOne (SYO) broth dilution colorimetric method. In addition, reference caspofungin MICs have so far proven to be unreliable. Candida species wild-type (WT) MIC distributions (for microorganisms in a species/drug combination with no detectable phenotypic resistance) were established for 6,007 Candida albicans, 186 C. dubliniensis, 3,188 C. glabrata complex, 119 C. guilliermondii, 493 C. krusei, 205 C. lusitaniae, 3,136 C. parapsilosis complex, and 1,016 C. tropicalis isolates. SYO MIC data gathered from 38 laboratories in Australia, Canada, Europe, Mexico, New Zealand, South Africa, and the United States were pooled to statistically define SYO ECVs. ECVs for anidulafungin, caspofungin, and micafungin encompassing ≥97.5% of the statistically modeled population were, respectively, 0.12, 0.25, and 0.06 µg/ml for C. albicans, 0.12, 0.25, and 0.03 µg/ml for C. glabrata complex, 4, 2, and 4 µg/ml for C. parapsilosis complex, 0.5, 0.25, and 0.06 µg/ml for C. tropicalis, 0.25, 1, and 0.25 µg/ml for C. krusei, 0.25, 1, and 0.12 µg/ml for C. lusitaniae, 4, 2, and 2 µg/ml for C. guilliermondii, and 0.25, 0.25, and 0.12 µg/ml for C. dubliniensis. Species-specific SYO ECVs for anidulafungin, caspofungin, and micafungin correctly classified 72 (88.9%), 74 (91.4%), 76 (93.8%), respectively, of 81 Candida isolates with identified fks mutations. SYO ECVs may aid in detecting non-WT isolates with reduced susceptibility to anidulafungin, micafungin, and especially caspofungin, since testing the susceptibilities of Candida spp. to caspofungin by reference methodologies is not recommended.
Assuntos
Antifúngicos/farmacologia , Candida/efeitos dos fármacos , Equinocandinas/farmacologia , Lipopeptídeos/farmacologia , Anidulafungina , Candida/genética , Caspofungina , Micafungina , Testes de Sensibilidade Microbiana , Mutação/genéticaRESUMO
FoxM1 is a member of the Forkhead family of transcription factors and is implicated in inducing cell proliferation and some forms of tumorigenesis. It binds promoter regions with a preference for tandem repeats of a consensus 'TAAACA' recognition sequence. The affinity of the isolated FoxM1 DNA-binding domain for this site is in the micromolar range, lower than observed for other Forkhead proteins. To explain these FoxM1 features, we determined the crystal structure of its DNA-binding domain in complex with a tandem recognition sequence. FoxM1 adopts the winged-helix fold, typical of the Forkhead family. Neither 'wing' of the fold however, makes significant contacts with the DNA, while the second, C-terminal, wing adopts an unusual ordered conformation across the back of the molecule. The lack of standard DNA-'wing' interactions may be a reason for FoxM1's relatively low affinity. The role of the 'wings' is possibly undertaken by other FoxM1 regions outside the DBD, that could interact with the target DNA directly or mediate interactions with other binding partners. Finally, we were unable to show a clear preference for tandem consensus site recognition in DNA-binding, transcription activation or bioinformatics analysis; FoxM1's moniker, 'Trident', is not supported by our data.
Assuntos
Fatores de Transcrição Forkhead/química , Regiões Promotoras Genéticas , Sequência de Bases , Sítios de Ligação , Sequência Consenso , DNA/química , Proteína Forkhead Box M1 , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Genoma Humano , Humanos , Modelos Moleculares , Mutação , Ligação Proteica , Multimerização Proteica , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Sequências de Repetição em Tandem , Transcrição GênicaRESUMO
Monocular elevation deficiency (MED) is characterized by unilateral limitation of supraductions, similar in adduction and abduction, in addition to hypotropia and ptosis. We describe a case of a 62-year-old woman with long-standing left ptosis who was initially operated with a frontal suspension technique. On subsequent examinations, a MED of that eye was found. The passive duction test was positive, so the inferior rectus was recessed. In addition, a modified Nishida technique was performed, consisting of the scleral anchorage of the superior edges of the rectus, medial and lateral, 12mm from the corneal limbus in the superonasal and superotemporal quadrants, respectively. Postoperatively, the left eye presented a minimal hypotropia of 3 PD. This modified technnique used here on the horizontal rectus muscles, turned out to be effective and safe for the correction of MED.
Assuntos
Blefaroptose , Estrabismo , Blefaroptose/cirurgia , Feminino , Humanos , Pessoa de Meia-Idade , Músculos Oculomotores/cirurgia , Procedimentos Cirúrgicos Oftalmológicos , Esclera , Estrabismo/cirurgiaRESUMO
Yeasts are the key microorganisms that transform grape juice into wine, and nitrogen is an essential nutrient able to affect yeast cell growth, fermentation kinetics and wine quality. In this work, we focused on the intra- and extracellular metabolomic changes of three aromatic amino acids (tryptophan, tyrosine, and phenylalanine) during alcoholic fermentation of two grape musts by two Saccharomyces cerevisiae strains and the sequential inoculation of Torulaspora delbrueckii with Saccharomyces cerevisiae. An UPLC-MS/MS method was used to monitor 33 metabolites, and 26 of them were detected in the extracellular samples and 8 were detected in the intracellular ones. The results indicate that the most intensive metabolomic changes occurred during the logarithm cellular growth phase and that pure S. cerevisiae fermentations produced higher amounts of N-acetyl derivatives of tryptophan and tyrosine and the off-odour molecule 2-aminoacetophenone. The sequentially inoculated fermentations showed a slower evolution and a higher production of metabolites linked to the well-known plant hormone indole acetic acid (auxin). Finally, the production of sulfonated tryptophol during must fermentation was confirmed, which also may explain the bitter taste of wines produced by Torulaspora delbrueckii co-fermentations, while sulfonated indole carboxylic acid was detected for the first time in such an experimental design.
RESUMO
Monocular elevation deficiency (MED) is characterized by unilateral limitation of supraductions, similar in adduction and abduction, in addition to hypotropia and ptosis. We describe a case of a 62-year-old woman with long-standing left ptosis who was initially operated with a frontal suspension technique. On subsequent examinations, a MED of that eye was found. The passive duction test was positive, so the inferior rectus was recessed. In addition, a modified Nishida technique was performed, consisting of the scleral anchorage of the superior edges of the rectus, medial and lateral, 12mm from the corneal limbus in the superonasal and superotemporal quadrants, respectively. Postoperatively, the left eye presented a minimal hypotropia of 3 PD. This modified technnique used here on the horizontal rectus muscles, turned out to be effective and safe for the correction of MED.
RESUMO
Yeast nitrogen metabolism produces metabolites, whose origin in wines has scarcely been studied, with an important biological and organoleptic role. The present work focuses on comparing three intracellular extraction methods in order to elucidate efficiency of extraction while measuring the effect of temperature upon the integrity of the compounds related to the metabolism of tryptophan and tyrosine by yeast. Two UHPLC/HRMS methods to measure 16 metabolites were developed and validated. The validation provided optimum values of LOD (7.4·10-6 to 0.1⯵gâ¯L-1), of LOQ (2·10-5 to 0.02⯵gâ¯L-1) of precision (11-0.5% RSD) and repeatability (12-0.5% RSD). The removal of interfering molecules enabled matrix effects to be kept at low levels. The results pointed out that the low-temperature methods were more effective, providing better precision for 16 metabolites. The high-temperature extraction method may yield false enhanced compounds concentrations since they originate in cell wall macromolecules degradation.
Assuntos
Fracionamento Químico/métodos , Saccharomyces cerevisiae/metabolismo , Triptofano/análise , Tirosina/análise , Cromatografia Líquida de Alta Pressão , Temperatura Baixa , Congelamento , Glicerol/química , Espectrometria de Massas , Nitrogênio/metabolismo , Triptofano/metabolismo , Tirosina/metabolismo , Vinho/análiseRESUMO
Tryptophan, phenylalanine, and tyrosine play an important role as nitrogen sources in yeast metabolism. They regulate biomass production and fermentation rate, and their catabolites contribute to wine health benefits and sensorial character through the yeast biotransformation of grape juice constitutes into biologically active and flavor-impacting components. A UHPLC-MS/MS method was applied to monitor 37 tryptophan/phenylalanine/tyrosine yeast metabolites both in extra- and intracellular extracts produced by the fermentation of two Saccharomyces cerevisiae strains and one Torulaspora delbrueckii. The results shed light on the intra- and extra-cellular metabolomic dynamics, by combining metabolic needs, stimuli, and signals. Among others, the results indicated (a) the production of 2-aminoacetophenone by yeasts, mainly by the two Saccharomyces cerevisiae; (b) the deactivation and/or detoxification of tryptophol via sulfonation reaction; and (c) the deacetylation of N-acetyl tryptophan ethyl ester and N-acetyl tyrosine ethyl ester by producing the corresponding ethyl esters.
Assuntos
Aminoácidos Aromáticos/metabolismo , Saccharomyces cerevisiae/metabolismo , Torulaspora/metabolismo , Aminoácidos Aromáticos/química , Cromatografia Líquida de Alta Pressão , Nitrogênio/metabolismo , Saccharomyces cerevisiae/química , Espectrometria de Massas em Tandem , Torulaspora/químicaRESUMO
X-linked adrenoleukodystrophy (X-ALD) belongs to a family of rare diseases due to inborn errors of metabolism. It has a wide spectrum of clinical manifestations that anaesthesiologists should recognise during the perioperative period (respiratory centre dysfunction, hypotonia, adrenal or hepatic failure, gastroesophageal reflux disease, osteopenia, seizures).The case is presented of a 42-year-old X-linked adrenoleukodystrophy female carrier, who underwent combined general and an ultrasound-guided interscalene brachial plexus block anaesthesia for shoulder arthroscopy. Induction was performed with thiopentone and fentanyl, and sevoflurane was used as inhaled maintenance agent. No events were recorded during the procedure. Her post-operative recovery was satisfactory and she was later discharged home.
Assuntos
Adrenoleucodistrofia/genética , Artroscopia , Bloqueio do Plexo Braquial/métodos , Heterozigoto , Ombro/cirurgia , Adulto , Anestesia Geral , Anestésicos , Feminino , Fentanila , Humanos , Sevoflurano , TiopentalAssuntos
Abscesso Abdominal/etiologia , Infecções Relacionadas a Cateter/etiologia , Corpos Estranhos/complicações , Pielonefrite/etiologia , Ureter , Cateterismo Urinário/efeitos adversos , Abscesso Abdominal/tratamento farmacológico , Abscesso Abdominal/cirurgia , Adulto , Falha de Equipamento , Feminino , Corpos Estranhos/diagnóstico por imagem , Corpos Estranhos/cirurgia , Humanos , Hidronefrose/etiologia , Insuficiência de Múltiplos Órgãos/etiologia , Esclerose Múltipla/complicações , Pielonefrite/tratamento farmacológico , Choque Séptico/etiologia , Tomografia Computadorizada por Raios X , Ureter/diagnóstico por imagem , Bexiga Urinaria Neurogênica/complicaçõesRESUMO
Hydroxytyrosol (HT) is a phenolic compound of recognized bioactivity that has been described in wines but little is known about its origin. This work demonstrates that yeast involved in wine making, i.e. Saccharomyces cerevisiae strains and the non-Saccharomyces Torulaspora delbrueckii, can synthesise HT, as this compound was identified in the intracellular media of three strains by means of a developed and validated UHPLC-HRMS method with LOQ and LOD of 0.108 and 0.035ngmL-1 respectively. Controlled fermentations were performed with different varieties of grapes (Corredera, Moscatel, Chardonnay, Palomino fino, Sauvignon Blanc, Vijiriega, and Tempranillo) and synthetic must. The Saccharomyces cerevisiae strain QA23 was the most efficient producer of HT from tested yeasts. On the other hand, the grape variety influences HT wine concentrations. Furthermore, the maximum concentration of HT is reached between the fourth and sixth day of fermentation. This work reveals that yeasts have a great potential for the production of HT.
Assuntos
Fermentação , Álcool Feniletílico/análogos & derivados , Saccharomyces cerevisiae/metabolismo , Torulaspora/metabolismo , Vinho/análise , Cromatografia Líquida de Alta Pressão , Álcool Feniletílico/análise , Álcool Feniletílico/metabolismo , VitisRESUMO
OBJECTIVE: The prevalence of Pneumocystis jirovecii colonization and its role in pulmonary disease remains unclear. PCR methods have shown an improved sensitivity in the detection of this fungus. It has been suggested that the PCR results be combined with another test such as IFA to create a diagnostic algorithm. METHODS: A multiplex nested-PCR procedure with a 16S rRNA gene as the internal amplification control was evaluated to determine the role of P. jirovecii in pulmonary disease. RESULTS: A 20% of the 199 bronchoalveolar lavage samples were PCR-positive, 13.5% samples were PCR-inhibited, and the rate of Pneumocystis-colonisation was 6.4%. The sensitivity, specificity, positive predictive value and negative predictive value of the nested-PCR were 100%, 93%, 70% and 100%, respectively. The sensitivity of the nested-PCR was higher than the current "gold standard" immunofluorescence assay (IFA) (p< 0.0001). PCR-negative and PCR-positive patients did not show any clinical or radiological differences in the medical variables studied. CONCLUSIONS: PCR could help the diagnosis of Pneumocystis pulmonary disease given the high negative predictive value of the technique. P. jirovecii DNA can frequently be detected in healthy population, so the analysis of the patient medical history is critical to make the correct clinical decision.
Assuntos
Pneumopatias/complicações , Pneumocystis carinii , Pneumonia por Pneumocystis/complicações , Pneumonia por Pneumocystis/diagnóstico , Reação em Cadeia da Polimerase/métodos , Idoso , Idoso de 80 Anos ou mais , Algoritmos , Líquido da Lavagem Broncoalveolar/microbiologia , DNA Bacteriano/genética , Feminino , Humanos , Pneumopatias/tratamento farmacológico , Pneumopatias/microbiologia , Masculino , Pessoa de Meia-Idade , Pneumocystis carinii/genética , Valor Preditivo dos Testes , RNA Bacteriano/genética , RNA Ribossômico 16S/genéticaRESUMO
Melatonin is a neurohormone involved in the regulation of circadian rhythms in humans. Evidence has recently been found of its occurrence in wines and its role in the winemaking process. The yeast Saccharomyces cerevisiae is consequently thought to be important in Melatonin synthesis, but limited data and reference texts are available on this synthetic pathway. This paper aims to elucidate whether the synthetic pathway of Melatonin in Saccharomyces and non-Saccharomyces strains involves these intermediates. To this end, seven commercial strains comprising Saccharomyces cerevisiae (Red Fruit, ES488, Lalvin QA23, Uvaferm BC, and Lalvin ICV GRE) and non-Saccharomyces (Torulaspora delbrueckii and Metschnikowia pulcherrima) were monitored, under controlled fermentation conditions, in synthetic must, for seven days. Samples were analysed using a UHPLC-HRMS system (Qexactive). Five out of the seven strains formed Melatonin during the fermentation process: three S. cerevisiae strains and the two non-Saccharomyces. Additionally, other compounds derived from l-tryptophan occurred during fermentation.
Assuntos
Fermentação/fisiologia , Melatonina/metabolismo , Saccharomyces cerevisiae/metabolismo , Triptofano/metabolismo , Vinho , Aromatizantes/isolamento & purificação , Aromatizantes/metabolismo , Melatonina/isolamento & purificação , Saccharomyces/metabolismo , Triptofano/isolamento & purificação , Vinho/análiseRESUMO
Excessive concentrations of vascular endothelial growth factor (VEGF) trigger angiogenesis, which causes complications such as the destabilization of atherosclerotic plaques and increased growth of tumors. This work focuses on the determination of the inhibitory activity of melatonin and other indolic related compounds on VEGF-induced VEGF receptor-2 (VEGFR-2) activation and an approximation to the molecular mechanism underlying the inhibition. Quantification of phosphorylated VEGFR-2 was measured by ELISA. Migration wound-healing assay was used to determine cell migration of human umbilical vein endothelial cells (HUVECs). This is the first time that melatonin, 3-indolacetic acid, 5-hydroxytryptophol, and serotonin are proved to significantly inhibit VEGF-induced VEGFR-2 activation in human umbilical vein endothelial cells and subsequent angiogenesis. 3-Indolacetic acid showed the highest inhibitory effect (IC50 value of 0.9704 mM), followed by 5-hydroxytryptophol (35% of inhibition at 0.1 mM), melatonin (30% of inhibition at 1 mM), and serotonin (24% of inhibition at 1 mM). An approximation to the molecular mechanism of the inhibition has been proposed, suggesting that indolic compounds might interact with the cell surface components of the endothelial membrane in a way that prevents VEGF from activating the receptor. Additionally, wound-healing assay revealed that exposure of HUVECs to melatonin and 3-indolacetic acid in the presence of VEGF significantly inhibited cell migration by 87% and 99%, respectively, after 24 h. These data demonstrate that melatonin, 3-indolacetic acid, 5-hydroxytryptophol, and serotonin would be good molecules for future exploitation as anti-VEGF signaling agents.
Assuntos
Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Indóis/farmacologia , Melatonina/farmacologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Inibidores da Angiogênese/farmacologia , Movimento Celular/efeitos dos fármacos , Células Cultivadas , Humanos , FosforilaçãoRESUMO
Anthocyanins are the major polyphenolic compounds in strawberry fruit responsible for its color. Due to their sensitivity, they are affected by food processing techniques such as fermentation that alters both their chemical composition and organoleptic properties. This work aims to evaluate the impact of different fermentation processes on individual anthocyanins compounds in strawberry wine and vinegar by UHPLC-MS/MS Q Exactive analysis. Nineteen, 18, and 14 anthocyanin compounds were identified in the strawberry initial substrate, strawberry wine, and strawberry vinegar, respectively. Four and 8 anthocyanin compounds were tentatively identified with high accuracy for the 1st time to be present in the beverages obtained by alcoholic fermentation and acetic fermentation of strawberry, respectively. Both, the total and the individual anthocyanin concentrations were decreased by both fermentation processes, affecting the alcoholic fermentation to a lesser extent (19%) than the acetic fermentation (91%). Indeed, several changes in color parameters have been assessed. The color of the wine and the vinegar made from strawberry changed during the fermentation process, varying from red to orange color, this fact is directly correlated with the decrease of anthocyanins compounds.
Assuntos
Antocianinas/química , Fragaria/química , Vinho/análise , Ácido Acético/análise , Bebidas/análise , Cor , Fermentação , Manipulação de Alimentos/métodos , Frutas/química , Espectrometria de Massas em TandemRESUMO
Overproduction of strawberry leads to food waste, as it is very perishable. Therefore, strategies to transform it into new products are appreciated. This research focuses on characterization of the nonanthocyanin phenolic content of a beverage obtained from strawberry by gluconic and acetic fermentation and subsequently monitored for 90 days of storage, at two temperatures. Sixty-four nonanthocyanin (poly)phenols were identified by high-resolution mass spectrometry (UHPLC coupled with linear trap quadrupole and Orbitrap mass analyzer) and, for the first time, four compounds were reported in beverages fermented from strawberry: aromadendrin hexoside, phloretin 2'-O-xylosyl glucoside, dihydroferulic acid 4-O-glucuronide, and kaempferol hexosyl hexoside. During the storage time the increase in protocatechuic acid content was 13 times and condensed tannins diminished, especially procyanidin trimer. Statistical analysis showed that the composition remains unchanged until day 15 of storage at room temperature (27-30 °C) and until day 30 under refrigerated conditions (4 °C).