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1.
J Orthop Sci ; 2024 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-38777662

RESUMO

BACKGROUND: To the best of our knowledge, no prior studies have identified any risk factors for subchondral insufficiency fractures of the medial tibial condyle. This study aimed to explain relationships between subchondral insufficiency fractures of the medial tibial condyle and the meniscus status, lower extremity alignment, or osteoporosis. METHODS: This retrospective study included 325 consecutive patients whose chief complaint is knee joint pain and who had visited one institution between April 2016 and March 2021, of which 70 patients (8 men and 62 women) who had suspected subchondral insufficiency fractures of the medial tibial condyle had undergone magnetic resonance imaging and radiographic examination. These patients were divided into two groups based on the results of their magnetic resonance imaging: the insufficiency fracture group included 46 patients who had subchondral insufficiency fractures of the medial tibial condyle and the nonfracture group included 24 patients without fractures. The meniscus injury and medial meniscus extrusion (MME) were evaluated by using magnetic resonance imaging. The Kellgren-Lawrence grade, the femorotibial angle, and the percent mechanical axis (%MA) were evaluated with the use of knee radiographs. T-scores were also measured by using dual-energy X-ray absorptiometry with a bone densitometer. RESULTS: MME were significantly larger and the %MA was significantly smaller in the insufficiency fracture group than that in the nonfracture group. The prevalence of medial meniscus injuries and pathological MME were higher in the insufficiency fracture group than those in the nonfracture group. The prevalence of varus knee and osteoporosis did not vary remarkably different between the two groups. CONCLUSION: The patients who had insufficiency fractures of the medial tibial condyle tended to have medial meniscus extrusion.

2.
Vet Sci ; 10(2)2023 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-36851463

RESUMO

Soluble guanylate cyclase (sGC) stimulator riociguat is a relatively novel therapeutic agent for pulmonary hypertension (PH) in human medicine. Riociguat induces endothelium-independent pulmonary artery (PA) relaxation by directly activating the sGC-cyclic guanosine-3',5'-monophosphate (cGMP) pathway in muscle cells. Although riociguat may be effective in the treatment of dogs with refractory PH, basic studies on its clinical application in veterinary medicine are lacking. The present study aimed to explore the effects of riociguat on the contractility of an isolated canine PA and the hemodynamics of dogs with acute PH. In an isolated endothelium-denuded canine PA, the effects of riociguat on endothelin (ET)-1-induced contraction and cGMP levels were investigated using the Magnus method and ELISA, respectively. The effect of riociguat on the hemodynamics of the thromboxane A2 analog U46619-induced PH model dog was examined by invasive catheterization. Riociguat increased cGMP levels and reduced ET-1-induced contraction of the isolated PA. Riociguat inhibited the U46619-induced elevation of PA pressure and pulmonary vascular resistance and increased cardiac output, but it had no effect on basal systemic blood pressure. These results demonstrate for the first time that riociguat can inhibit the elevation of PA pressure through PA relaxation via an endothelium-independent increase in cGMP in dogs with PH.

3.
JTCVS Open ; 11: 49-58, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36172424

RESUMO

Objective: Postoperative atrial fibrillation (POAF) after cardiac surgery is associated with increased mortality. The efficacy of landiolol hydrochloride for POAF prevention after coronary artery bypass grafting procedure and valve surgery has been reported. However, little evidence is available on its role in POAF prevention after aortic root, ascending aorta, and aortic arch surgery. This study aimed to determine the association between intravenous landiolol and the incidence of POAF after these aortic surgeries. Methods: We included 358 consecutive adult patients without preoperative atrial fibrillation who underwent aortic root, ascending aorta, and aortic arch surgery between January 1, 2011, and December 31, 2018, at our institution. The therapeutic influence of landiolol in preventing POAF was estimated by propensity score-matched analysis (n = 222). The primary end point was the incidence of POAF within 72 hours after surgery. The secondary end points included adverse clinical events such as 30-day mortality and symptomatic cerebral infarction. Results: The median age of the cohort was 72 years, 68.5% were men, and 46.4% received postoperative oral or transdermal ß-blockers. After minimizing differences in patient background by propensity score matching, the incidence of POAF in the landiolol group was significantly lower than that in the reference group (18.9% vs 38.7%; P = .002). Landiolol use was associated with reduced incidence of POAF (odds ratio, 0.39; 95% CI, 0.21 to -0.72; P = .003). There were no significant differences in secondary end points. Conclusions: Intravenous landiolol was associated with a lower incidence of POAF after aortic root, ascending aorta, and aortic arch surgery.

4.
J Cardiol Cases ; 24(2): 72-74, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34354781

RESUMO

An 11-year-old boy with no medical or family history was diagnosed with Stanford type B acute aortic dissection. Although a conservative treatment approach was adopted, deep sedation was required to keep him still during computed tomography. It revealed enlargement of the false lumen of the descending aorta, bilateral pleural effusion, and atelectasis. Thus, he underwent descending aortic replacement. After amelioration of perioperative rhabdomyolysis, he was discharged post-recovery. Since there have been no clinical guidelines for management of pediatric aortic dissection, it was difficult to decide between surgical and conservative approaches. Considering difficulty of mild sedation in children, if conservative approaches seem to be problematic, an early surgical approach with aortic replacement is sometimes necessary. .

5.
J Anesth ; 35(5): 611-616, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34086138

RESUMO

PURPOSE: An electromyographic (EMG) tube is sometimes used for vagal nerve monitoring during neurosurgery. Some characteristics of an EMG tube are different from those of a normal endotracheal tube. Although postoperative laryngeal edema (PLE) may occur and reintubation may be required in some patients in whom an EMG tube is used, its relevance to these events has not been investigated in detail. Our goal was to determine the relevance of an EMG tube to the development of PLE and the need for reintubation. METHODS: A retrospective study was conducted in 900 patients after neurosurgery from 2012 to 2018. Severe PLE occurrence or the requirement for postoperative reintubation were compared between the EMG tube (E) group and the Normal tube (N) group, using a propensity score (PS) matching analysis RESULTS: After PS matching, severe PLE incidence (n = 2/20, 10.0%) in the E group was significantly higher than that (0/80, 0%) in the N group. There was no significant difference in the incidence of reintubation between the E group (1/20, 5.0%) and the N group (0/80, 0%). CONCLUSION: Electromyographic (EMG) tube use was significantly associated with higher incidence of severe PLE.


Assuntos
Edema Laríngeo , Neurocirurgia , Eletromiografia , Humanos , Intubação Intratraqueal/efeitos adversos , Edema Laríngeo/diagnóstico , Edema Laríngeo/etiologia , Estudos Retrospectivos
6.
Jpn J Clin Oncol ; 51(3): 354-362, 2021 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-33173939

RESUMO

OBJECTIVE: This phase 2 study examined the efficacy and safety of tolvaptan, an aquaretic drug, in the treatment of ascites associated with cancer. METHODS: In the dose-escalation phase, oral tolvaptan was initiated at a dose of 3.75 mg/day, and the dose was increased daily to 7.5, 15 and 30 mg/day. Dose escalation was terminated once the increase from baseline in the daily urine volume reached 500 ml, at which point the patient proceeded to the maintenance phase of 5-7 days. Improvement of ascites was determined primarily by reduction in body weight and ascitic fluid volume. RESULTS: The mean change from baseline in body weight was maintained below 0 kg throughout the study. The mean change (±standard deviation) from baseline in ascitic fluid volume at the end of treatment (EOT) was 237.45 ± 868.14 ml in 33 evaluable patients. Although an increase from baseline in ascitic fluid volume at the EOT was observed in 23 of 33 patients (maximum: 1589.3 ml, minimum: 3.83 ml), a reduction in ascitic fluid volume was observed in the remaining 10 patients (maximum: -2304.3 ml, minimum: -27.5 ml). The common treatment-emergent adverse events included vomiting (5 of 43 patients, 11.6%), abdominal distension, constipation, thirst, blood osmolarity increased and renal impairment (3 of 43 patients, 7.0% each). CONCLUSIONS: Tolvaptan seemed to have no definitive effect on reducing ascites; however, it might be effective in at least some cancer patients. No new safety concerns were identified at doses of 3.75-30 mg/day.


Assuntos
Ascite/tratamento farmacológico , Tolvaptan/efeitos adversos , Tolvaptan/uso terapêutico , Idoso , Antagonistas dos Receptores de Hormônios Antidiuréticos/efeitos adversos , Antagonistas dos Receptores de Hormônios Antidiuréticos/uso terapêutico , Ascite/patologia , Peso Corporal/efeitos dos fármacos , Feminino , Humanos , Masculino , Resultado do Tratamento
7.
Appl Environ Microbiol ; 84(1)2018 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-29079616

RESUMO

In soy sauce manufacturing, Candida versatilis plays a role in the production of volatile flavor compounds, such as volatile phenols, but limited accessible information on its genome has prevented further investigation regarding aroma production and breeding. Although the draft genome sequence data of two strains of C. versatilis have recently been reported, these strains are not similar to each other. Here, we reassess the draft genome sequence data for strain t-1, which was originally reported to be C. versatilis, and conclude that strain t-1 is most probably not C. versatilis but a gamete of hybrid Zygosaccharomyces rouxii Phylogenetic analysis of the D1/D2 region of the 26S ribosomal DNA (rDNA) sequence indicated that strain t-1 is more similar to the genus Zygosaccharomyces than to C. versatilis Moreover, we found that the genome of strain t-1 is composed of haploid genome content and divided into two regions that show approximately 100% identity with the T or P subgenome derived from the natural hybrid Zygosaccharomyces rouxii, such as NBRC110957 and NBRC1876. We also found a chromosome crossing-over signature in the scaffolds of strain t-1. These results suggest that strain t-1 is a gamete of the hybrid Z. rouxii, generated by either meiosis or chromosome loss following reciprocal translocation between the T and P subgenomes. Although it is unclear why strain t-1 was misidentified as C. versatilis, the genome of strain t-1 has broad implications for considering the evolutionary fate of an allodiploid.IMPORTANCE In yeast, crossing between different species sometimes leads to interspecies hybrids. The hybrid generally cannot produce viable spores because dissimilarity of parental genomes prevents normal chromosome segregation during meiotic division, leading to a dead end. Thus, only a few natural cases of homoploid hybrid speciation, which requires mating between 1n gametes of hybrids, have been described. However, a recent study provided strong evidence that homoploid hybrid speciation is initiated in natural populations of the budding yeast, suggesting the potential presence of viable 1n gametes of hybrids. The significance of our study is finding that the strain t-1, which had been misidentified as Candida versatilis, is a viable 1n gamete derived from hybrid Zygosaccharomyces rouxii.


Assuntos
Candida/fisiologia , DNA Fúngico/análise , RNA Ribossômico/análise , Zygosaccharomyces/classificação , Zygosaccharomyces/fisiologia , Evolução Biológica , Candida/classificação , Hibridização Genética , Filogenia
8.
Appl Environ Microbiol ; 83(21)2017 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-28842546

RESUMO

The mechanism of whole-genome duplication (WGD) in yeast has been intensively studied because it has a large impact on yeast evolution. WGD has shaped the genomic architecture of modern Saccharomyces cerevisiae; however, the mechanism for restoring fertility after interspecies hybridization, which would be involved in the process of WGD, has not been thoroughly elucidated. In this study, we obtained a draft genome sequence of the salt-tolerant yeast Zygosaccharomyces rouxii NBRC110957 and revealed that it is a hybrid lineage of Z. rouxii (allodiploid) with two subgenomes equivalent to NBRC1876. Because this allodiploid yeast can mate with other allodiploid strains and form spores, it can be a good model of restoring fertility after interspecies hybridization. We observed that NBRC110957 and NBRC1876 contain six mating-type-like (MTL) loci. There are no large deletions or deleterious mutations in MTL loci, except for several-base-pair deletions in the X region in certain MTL loci. We also assigned only one mating-type (MAT) locus that exclusively determines mating types from six MTL loci. These results suggest that it is possible to recover mating competence regardless of whether cells lose one MAT locus through random gene loss by mitotically dividing after interspecies hybridization. Moreover, we propose that perturbation of gene expression and substantial breakdown of MAT heterozygosity caused by chromosomal rearrangement at MTL loci play roles in restoring the mating competence of allodiploids. This scenario can provide a mechanism for restoring fertility after interspecies hybridization that is compatible with random gene loss models and suggests genomic plasticity during WGD in yeast.IMPORTANCE A whole-genome duplication occurred in an ancestor of the baker's yeast Saccharomyces cerevisiae The origins of this complex and multifaceted process, which requires intra- or interspecies hybridization followed by dysfunction of one mating-type (MAT) locus to regain mating competence, has not been thoroughly elucidated. In this study, we provide a mechanism for regaining fertility in an interspecies hybrid, Zygosaccharomyces rouxii The draft genome sequence analysis and mating test showed that the Z. rouxii strain used in this study is an intact interspecies hybrid, suggesting that it is possible to recover fertility regardless of whether cells lose one MAT locus.


Assuntos
Hibridização Genética , Zygosaccharomyces/fisiologia , Diploide , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Genes Fúngicos Tipo Acasalamento , Zygosaccharomyces/genética
9.
J Biosci Bioeng ; 123(3): 333-341, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27865643

RESUMO

4-Hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone (HEMF) is considered a key flavor compound in soy sauce. The compound has a caramel-like aroma and several important physiological activities, such as strong antioxidant activity. Here, we report the identification and characterization of an enzyme involved in the biosynthesis of HEMF in yeast. We fractionated yeast cell-free extract from Saccharomyces cerevisiae using column chromatography and partially purified a fraction with HEMF-forming activity. Peptide mass fingerprinting analysis showed that the partially purified fraction contains aldehyde reductase encoded by YNL134C. This reductase shares low sequence identity with enone oxidoreductase, which is responsible for the formation of 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) and HEMF in plants. YNL134C was expressed heterologously in Escherichia coli, and the purified protein catalyzed the formation of HEMF from the mixture of Maillard reaction products, acetaldehydes, and NADPH. Multicopy expression in S. cerevisiae resulted in increased HEMF productivity, and gene knockout of YNL134C in S. cerevisiae resulted in decreased HEMF productivity. These data suggest that the translation product of YNL134C is the HEMF-producing enzyme in yeast. Detailed analyses of an intermediate in the enzymatic reaction mixture revealed that HEMF is synthesized from (2E)-2-ethylidene-4-hydroxy-5-methyl-3(2H)-furanone, which formed via Knoevenagel condensation between the acetaldehyde and 4-hydroxy-5-methyl-3(2H)-furanone derived from the Maillard reaction based on ribose and glycine, by YNL134Cp in an NADPH dependent manner. Overall, this study shed light on the molecular basis for the improvement of soy sauce flavor and the biotechnological production of HEMF.


Assuntos
Furanos/metabolismo , Oxirredutases/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/metabolismo , Acetaldeído/metabolismo , Aldeído Redutase/deficiência , Aldeído Redutase/genética , Aldeído Redutase/isolamento & purificação , Aldeído Redutase/metabolismo , Extratos Celulares , Escherichia coli/genética , Aromatizantes/química , Glicina/metabolismo , NADP/metabolismo , Oxirredutases/deficiência , Oxirredutases/genética , Oxirredutases/isolamento & purificação , Ribose/metabolismo , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/isolamento & purificação , Alimentos de Soja
10.
Appl Microbiol Biotechnol ; 99(18): 7685-97, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25982000

RESUMO

Soy sauce yeast Zygosaccharomyces rouxii plays a central role in the production of flavor compounds in soy sauce, while the flor-forming strain spoils its quality by producing 2-methylpropanoic acid, 2-methylbutanoic acid, and 3-methylbutanoic acid, which have an unpleasant odor. To investigate the relationship between flor formation and unpleasant odor, we measured the volatile compounds that accumulated under various growth conditions. As a result, marked amounts of 2-methylpropanoic acid, 2-methylbutanoic acid, or 3-methylbutanoic acid accumulated in synthetic medium containing valine, isoleucine, or leucine, respectively, under aerobic growth conditions. These results implied that the unpleasant compounds were produced from their corresponding branched chain amino acid (BCAA) when the cell was placed under aerobic condition through flor formation. The first step in BCAA catabolism and the last step in BCAA anabolism are both catalyzed by a BCAA transaminase. A mutant lacking the BCAA transaminase gene, BAT1, resulted in valine and isoleucine auxotrophy, while a mutant lacking both BAT1 and the α-aminoadipate aminotransferase gene, ARO8, resulted in valine, isoleucine, and leucine auxotrophy. Although the bat1∆ aro8∆ double mutant formed flor similarly to the wild-type strain, the mutant exhibited less unpleasant odor generation. These results suggest that the interconversion between 4-methyl-2-oxopentanoate and leucine is catalyzed by both Bat1p and Aro8p in Z. rouxii. Taken together, these results indicate that flor formation is not seemed to be directly linked to unpleasant odor generation. These findings encourage us to breed flor-forming yeasts without an unpleasant odor.


Assuntos
2-Aminoadipato Transaminase/metabolismo , Odorantes , Transaminases/metabolismo , Compostos Orgânicos Voláteis/metabolismo , Zygosaccharomyces/enzimologia , Zygosaccharomyces/metabolismo , 2-Aminoadipato Transaminase/genética , Aerobiose , Meios de Cultura/química , Deleção de Genes , Transaminases/genética , Zygosaccharomyces/genética , Zygosaccharomyces/crescimento & desenvolvimento
11.
J Biosci Bioeng ; 120(4): 432-7, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25735923

RESUMO

In this study, we established stable cell lines producing 1.5 mg/mL recombinant human prothrombin in 400-L fed-batch culture, using CHO DG44 cells as a host cell line. And we also established a recombinant human α-thrombin purification process that produces a purified product suitable for use as a biopharmaceutical, by using recombinant ecarin from CHO DG44 cells, achieving a total yield of approximately 27% of prothrombin in culture medium. The establishment of stable cell lines with high expression levels, long-term passage stability and satisfactory scale-up are essential to ensure the stable supply of biopharmaceuticals. Furthermore, biopharmaceuticals must be of high quality to assure safety and effectiveness in target applications. We had previously reported that recombinant human prethrombin-2 expression level in a stable cell line established using the mouse myeloma cells, Sp2/0-Ag14, reached 200 µg/mL using animal-free materials in 50-L fed-batch culture. However, the productivity was insufficient to completely replace α-thrombin in human plasma preparations. By employing CHO DG44 cells as a host cell line, we had established a stable cell line and achieved significant improvements in quality, productivity of recombinant human α-thrombin manufacture suitable for use as a biopharmaceutical.


Assuntos
Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/isolamento & purificação , Trombina/biossíntese , Trombina/isolamento & purificação , Animais , Técnicas de Cultura Celular por Lotes , Células CHO , Cricetinae , Cricetulus , Meios de Cultura/química , Endopeptidases/genética , Endopeptidases/metabolismo , Humanos , Protrombina/análise , Protrombina/biossíntese , Proteínas Recombinantes/uso terapêutico , Trombina/uso terapêutico
12.
Mol Microbiol ; 96(1): 14-27, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25588312

RESUMO

Hydrophobins are amphipathic proteins secreted by filamentous fungi. When the industrial fungus Aspergillus oryzae is grown in a liquid medium containing the polyester polybutylene succinate co-adipate (PBSA), it produces RolA, a hydrophobin, and CutL1, a PBSA-degrading cutinase. Secreted RolA attaches to the surface of the PBSA particles and recruits CutL1, which then condenses on the particles and stimulates the hydrolysis of PBSA. Here, we identified amino acid residues that are required for the RolA-CutL1 interaction by using site-directed mutagenesis. We quantitatively analyzed kinetic profiles of the interactions between RolA variants and CutL1 variants by using a quartz crystal microbalance (QCM). The QCM analyses revealed that Asp142, Asp171 and Glu31, located on the hydrophilic molecular surface of CutL1, and His32 and Lys34, located in the N-terminus of RolA, play crucial roles in the RolA-CutL1 interaction via ionic interactions. RolA immobilized on a QCM electrode strongly interacted with CutL1 (K(D) = 6.5 nM); however, RolA with CutL1 variants, or RolA variants with CutL1, showed markedly larger KD values, particularly in the interaction between the double variant RolA-H32S/K34S and the triple variant CutL1-E31S/D142S/D171S (K(D) = 78.0 nM). We discuss a molecular prototype model of hydrophobin-based enzyme recruitment at the solid-water interface.


Assuntos
Aminoácidos Acídicos/metabolismo , Aspergillus oryzae/genética , Aspergillus oryzae/metabolismo , Hidrolases de Éster Carboxílico/metabolismo , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Sequência de Aminoácidos , Hidrolases de Éster Carboxílico/química , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Interações Hidrofóbicas e Hidrofílicas , Íons , Modelos Moleculares , Mutagênese Sítio-Dirigida , Poliésteres/metabolismo , Polímeros/metabolismo , Técnicas de Microbalança de Cristal de Quartzo
13.
Appl Environ Microbiol ; 81(1): 453-60, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25362059

RESUMO

4-Hydroxy-2 (or 5)-ethyl-5 (or 2)-methyl-3(2H)-furanone (HEMF) is an important flavor compound that contributes to the sensory properties of many natural products, particularly soy sauce and soybean paste. The compound exhibits a caramel-like aroma and several important physiological activities, such as strong antioxidant activity. HEMF is produced by yeast species in soy sauce manufacturing; however, the enzymes involved in HEMF production remain unknown, hindering efforts to breed yeasts with high-level HEMF production. In this study, we identified high-level HEMF-producing mutants among a Saccharomyces cerevisiae gene deletion mutant collection. Fourteen deletion mutants were screened as high-level HEMF-producing mutants, and the ADH1 gene deletion mutant (adh1Δ) exhibited the maximum HEMF production capacity. Further investigations of the adh1Δ mutant implied that acetaldehyde accumulation contributes to HEMF production, agreeing with previous findings. Therefore, acetaldehyde might be a precursor for HEMF. The ADH1 gene deletion mutant of Zygosaccharomyces rouxii, which is the dominant strain of yeast found during soy sauce fermentation, also produces HEMF effectively, suggesting that acetaldehyde accumulation might be a benchmark for breeding industrial yeasts with excellent HEMF production abilities.


Assuntos
Furanos/metabolismo , Deleção de Genes , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Acetaldeído/metabolismo , Álcool Desidrogenase/deficiência , Antioxidantes/metabolismo , Aromatizantes/metabolismo , Programas de Rastreamento , Proteínas de Saccharomyces cerevisiae
14.
Biosci Biotechnol Biochem ; 78(10): 1693-9, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25273133

RESUMO

Hydrophobins are amphipathic secretory proteins with eight conserved cysteine residues and are ubiquitous among filamentous fungi. The Cys3-Cys4 and Cys7-Cys8 loops of hydrophobins are thought to form hydrophobic segments involved in adsorption of hydrophobins on hydrophobic surfaces. When the fungus Aspergillus oryzae is grown in a liquid medium containing the polyester polybutylene succinate-co-adipate (PBSA), A. oryzae produces hydrophobin RolA, which attaches to PBSA. Here, we analyzed the kinetics of RolA adsorption on PBSA by using a PBSA pull-down assay and a quartz crystal microbalance (QCM) with PBSA-coated electrodes. We constructed RolA mutants in which hydrophobic amino acids in the two loops were replaced with serine, and we examined the kinetics of mutant adsorption on PBSA. QCM analysis revealed that mutants with replacements in the Cys7-Cys8 loop had lower affinity than wild-type RolA for PBSA, suggesting that this loop is involved in RolA adsorption on PBSA.


Assuntos
Aminoácidos , Aspergillus oryzae , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Polímeros/química , Adsorção , Sequência de Aminoácidos , Proteínas Fúngicas/genética , Microesferas , Dados de Sequência Molecular , Mutação , Ligação Proteica
15.
Masui ; 63(3): 309-14, 2014 Mar.
Artigo em Japonês | MEDLINE | ID: mdl-24724441

RESUMO

BACKGROUND: Chronic hemodialysis patients undergoing surgery require intensive care. The purpose of this study was to determine the incidence and risk factors of emergency treatment before the planned elective postoperative dialysis. METHODS: One hundred and ten dialysis patients undergoing elective noncardiac surgery were retrospectively analyzed. RESULTS: Emergency treatment was performed in 31 (28.1%) of the 110 patients, including one patient with emergency dialysis, because of hyperkalemia (n = 27) or metabolic acidosis (n = 4). Receiving operating characteristic curve analysis showed a cutoff value for serum potassium concentration < 4.0 mEq x l(-1) estimated by maximizing the Youden index. We could not find other perioperative data as significant risk factors for emergency treatment. CONCLUSIONS: About 30 percent of dialysis patients aftet elective noncardiac surgery required emergency treatment. The highest risk was high serum potassium concentrations after induction of anesthesia Preoperative serum potassium concentrations should be kept below 4.0 mEq x l(-1) to avoid postoperative emergency treatment.


Assuntos
Diálise , Procedimentos Cirúrgicos Eletivos , Serviços Médicos de Emergência/estatística & dados numéricos , Hiperpotassemia/epidemiologia , Hiperpotassemia/prevenção & controle , Cuidados Pós-Operatórios/estatística & dados numéricos , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/prevenção & controle , Procedimentos Cirúrgicos Operatórios , Idoso , Cloreto de Cálcio/administração & dosagem , Feminino , Humanos , Hiperpotassemia/terapia , Incidência , Unidades de Terapia Intensiva/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Assistência Perioperatória , Complicações Pós-Operatórias/terapia , Estudos Retrospectivos , Fatores de Risco , Bicarbonato de Sódio/administração & dosagem , Fatores de Tempo
16.
Clin Neuropathol ; 33(1): 68-75, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24216149

RESUMO

OBJECTIVE: Pathogenesis of pseudohypertrophy of the inferior olivary nucleus (PH-IO) was analyzed based on immunohistochemical study. METHODS: Immunostained medullas with PH-IO were observed with confocal laser microscopy. RESULTS: αB-crystallin (αBC) was frequently expressed in the neurons and co-localized with microtubule-associated protein 2 (MAP2). The neurons were occasionally positive for SMI-31. αBC and SMI-31 were co-localized in some neurons. Synaptophysin (SYP)-immunoreactive dots were present around MAP2-positive hypertrophic neurons and hypertrophic thick neurites. Periphery-stained Lys-Asp-Glu-Leu (KDEL)-positive neurons were shown. Central chromatolytic neurons were found with Klüver-Barrera staining, which indicated that the rough endoplasmic reticulum (ER) was distributed to the periphery of the cytoplasm. CONCLUSIONS: αBC prevents microtubule disassembly and phosphorylation of the neurofilaments under stressful conditions. Our results indicated that αBC protected microtubules and neurofilaments in PH-IO. The retrograde transport of KDEL receptors from the Golgi complex to the ER is increased under stressful conditions. We considered that KDEL receptors were retro-transported to ER, and then the ER containing KDEL receptors was distributed to the periphery of the cytoplasm. PH-IO showed various immunohistochemical changes due to trans-synaptic degeneration.


Assuntos
Núcleo Olivar/metabolismo , Núcleo Olivar/patologia , Retículo Endoplasmático/fisiologia , Complexo de Golgi/fisiologia , Humanos , Hipertrofia/etiologia , Hipertrofia/metabolismo , Hipertrofia/patologia , Microscopia Confocal , Receptores de Peptídeos/metabolismo , Degeneração Retrógrada/complicações , Degeneração Retrógrada/metabolismo , Degeneração Retrógrada/patologia , Cadeia B de alfa-Cristalina/metabolismo
17.
Genetics ; 195(2): 393-405, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23893487

RESUMO

Copy number variations (CNVs) contribute to the adaptation process in two possible ways. First, they may have a direct role, in which a certain number of copies often provide a selective advantage. Second, CNVs can also indirectly contribute to adaptation because a higher copy number increases the so-called "mutational target size." In this study, we show that the copy number amplification of FLO11D in the osmotolerant yeast Zygosaccharomyces rouxii promotes its further adaptation to a flor-formative environment, such as osmostress static culture conditions. We demonstrate that a gene, which was identified as FLO11D, is responsible for flor formation and that its expression is induced by osmostress under glucose-free conditions, which confer unique characteristics to Z. rouxii, such as osmostress-dependent flor formation. This organism possesses zero to three copies of FLO11D, and it appears likely that the FLO11D copy number increased in a branch of the Z. rouxii tree. The cellular hydrophobicity correlates with the FLO11D copy number, and the strain with a higher copy number of FLO11D exhibits a fitness advantage compared to a reference strain under osmostress static culture conditions. Our data indicate that the FLO gene-related system in Z. rouxii has evolved remarkably to adapt to osmostress environments.


Assuntos
Variações do Número de Cópias de DNA/genética , Glicoproteínas de Membrana/genética , Pressão Osmótica , Proteínas de Saccharomyces cerevisiae/genética , Zygosaccharomyces/genética , Sequência de Aminoácidos , Biofilmes/crescimento & desenvolvimento , Evolução Molecular , Regulação Fúngica da Expressão Gênica , Mutação , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/fisiologia , Zygosaccharomyces/fisiologia
18.
Masui ; 62(4): 477-80, 2013 Apr.
Artigo em Japonês | MEDLINE | ID: mdl-23697207

RESUMO

BACKGROUND: Transportable capnometers(EMMA) can be useful in the emergency department or Rapid Response System. Before EMMA can be implemented, it must be compared with currently employed capnography methods. Methods : The concentration of CO2 in a reference gas was measured by two EMMA machines and a side-stream capnometer (CAPNOX ), respectively. Next, Etco2 in twelve patients under general anesthesia was measured by both EMMA machines and the side-stream capnometer, respectively. Results were analyzed using Pearson's correlation coefficient and the Bland-Altman plot. Results : With regard to the reference gas ([CO2] of 38 mmHg), the EMMA machines reported CO, concentrations of 37.2 mmHg and 35 mmHg, and the capnometer reported 38 mmHg. For the 12 anesthetized patients, 47 Etco2 readings were taken. Pearson's correlation coefficient between the first EMMA machine and the capnometer was 0.98 (P<0.0001, bias 3.6 mmHg, 95% limits of agreement 1.3-5.9mmHg) and between the second EMMA machine and the capnometer was 0.99 (P<0.0001, bias 0.85 mmHg, 95% limits of agreement-0.7-2.4 mmHg). CONCLUSIONS: In patients under general anesthesia, EMMA measured Etco2 within 4 mmHg of side-stream capnography, indicating sufficient accuracy for clinical use. At the same time, discrepancies in readings between individual machines must be taken into consideration.


Assuntos
Capnografia/instrumentação , Monitorização Fisiológica/instrumentação , Capnografia/normas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
19.
Pathol Int ; 63(4): 206-13, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23692421

RESUMO

The histologic characteristics of air space enlargement with fibrosis (AEF) are compared with usual interstitial pneumonia (UIP), nonspecific interstitial pneumonia (NSIP) and centrilobular emphysema (CLE) to determine similarities and differences. Lung specimens from 39 patients were studied; 9 with AEF, 13 with UIP and 5 with CLE identified in lobectomy specimens for cancer and 12 NSIP cases identified on surgical lung biopsies. We determined the characteristics of cystic structures (i.e. abnormal airspace), degree of inflammation and severity of pneumocyte injury semi-quantitatively. In AEF, the wall thickness of the cystic lesions (0.8 mm) was thinner than in UIP (2.1 mm) and thicker than in CLE (0.07 mm). The degree of inflammation and granulation tissue were milder in AEF than in UIP and NSIP and CLE showed milder inflammatory cells than AEF. As for pneumocyte injury, AEF had fewer erosions (0.1/case) and fewer ubiquitin-positive pneumocytes than UIP (4.8 cells/slide) and NSIP (9.8 cells/slide). Our data suggested that the histological characteristics of AEF differed significantly from UIP, NSIP and CLE.


Assuntos
Doenças Pulmonares Intersticiais/patologia , Alvéolos Pulmonares/patologia , Enfisema Pulmonar/patologia , Fibrose Pulmonar/patologia , Idoso , Células Epiteliais Alveolares/patologia , Biomarcadores/metabolismo , Biópsia , Feminino , Tecido de Granulação/metabolismo , Tecido de Granulação/patologia , Humanos , Inflamação , Doenças Pulmonares Intersticiais/metabolismo , Masculino , Pessoa de Meia-Idade , Pneumonectomia , Alvéolos Pulmonares/metabolismo , Enfisema Pulmonar/metabolismo , Fibrose Pulmonar/metabolismo , Fumar/efeitos adversos , Tabagismo/metabolismo , Tabagismo/patologia
20.
PLoS One ; 8(4): e62121, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23614024

RESUMO

We investigated sex chromosome diversity in Zygosaccharomyces rouxii (Z. rouxii). In the current study, we show that the organization of the mating-type (MAT) locus is highly variable in the Z. rouxii population, indicating the MAT, HML, and HMR loci are translocation hotspots. Although NBRC1130 and CBS732 were originally two stocks of the type strain of the species, only NBRC1130 retains the original karyotype. A reciprocal translocation between the MAT and HMR loci appears to have occurred during the early passage culture of CBS732, which was used for genome sequencing. In NBRC1733, NBRC0686, NBRC0740 and NBRC1053, the terminal region of the chromosome containing the HMR locus was replaced with the chromosomal region to the left of the MAT or HML loci. The translocation events found in NBRC1733, NBRC0686, NBRC0740, and NBRC1053 were reconstructed under our experimental conditions using the DA2 background, and the reconstruction suggests that the frequency of this type of translocation is approximately 10(-7). These results suggest that the MAT and MAT-like loci were the susceptible regions in the genome, and the diversity of mating-type chromosome structures in Z. rouxii was caused by ectopic exchanges between MAT-like loci.


Assuntos
DNA Fúngico/genética , Zygosaccharomyces/genética , Mapeamento Cromossômico , Estruturas Cromossômicas
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