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1.
Gerontology ; 67(5): 517-524, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33596585

RESUMO

INTRODUCTION: Reduced tongue pressure is one of the causes of dysphagia. The purpose of this study was to investigate the relationship between dental findings and tongue pressure, and whether prosthetic treatment prevents reduced tongue pressure. METHODS: This was a cross-sectional study. Participants were 745 community-dwelling adults and elderly persons in the Goto Islands in Nagasaki, who underwent a health checkup for residents in 2015 and 2016. Data were collected on gender; age; grip strength; hemoglobin; Creatinine (Cr); glycated hemoglobin (HbA1c); history of stroke; smoking, drinking, exercise, and walking habits; number of teeth; wearing of removable dentures; functional units of natural teeth (n-FTUs), fixed prostheses (nif-FTUs), and removable dentures (t-FTUs); and tongue pressure. The associations between each variable and tongue pressure were examined using multiple regression analysis. Next, those with 3 or fewer n-FTUs were selected, and differences in tongue pressure were compared between those with 3 or fewer nif-FTUs and those with 4 or more nif-FTUs, using a propensity score matching method. RESULTS: Male gender, weak grip strength, low HbA1c, no drinking, and a low number of teeth were independent factors significantly associated with lower tongue pressure. Among participants with 3 or fewer n-FTUs, the 43 with 4 or more nif-FTUs showed significantly higher tongue pressure than the 43 with 3 or fewer nif-FTUs after propensity score matching, although the number of t-FTUs was not associated with tongue pressure. DISCUSSION/CONCLUSION: Tooth loss was significantly associated with lower tongue pressure. It was suggested that fixed prosthesis treatment might prevent the reduction of tongue pressure, but removable dentures did not have such an effect.


Assuntos
Vida Independente , Língua , Idoso , Estudos Transversais , Humanos , Japão/epidemiologia , Masculino , Pressão
2.
Asian J Endosc Surg ; 14(2): 258-261, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32790052

RESUMO

Traumatic diaphragmatic hernia is a serious complication of blunt trauma to the abdomen or thorax. Although traumatic diaphragmatic hernia is treated with surgical repair, a laparoscopic approach is infrequently employed. Here we present the case of a 66-year-old man with a bruise on the left side of his back. CT revealed a left pneumothorax and left rib fractures. He was urgently hospitalized and relieved with conservative treatment. However, on day 4 of hospitalization, an incarcerated diaphragmatic hernia containing the transverse colon was observed on CT. The herniated viscera of the abdominal cavity were reduced laparoscopically, and the hernial orifice was repaired with direct closure. One-lung ventilation was used to limit the movement of the affected diaphragm, enabling effective laparoscopic suturing. The patient had an uneventful recovery period and was discharged 8 days postoperatively. The absence of diaphragmatic herniation recurrence was confirmed 6 months after surgery.


Assuntos
Hérnia Diafragmática Traumática , Laparoscopia , Ferimentos não Penetrantes , Idoso , Colo , Diafragma , Hérnia Diafragmática Traumática/diagnóstico por imagem , Hérnia Diafragmática Traumática/etiologia , Hérnia Diafragmática Traumática/cirurgia , Humanos , Masculino , Ferimentos não Penetrantes/complicações , Ferimentos não Penetrantes/cirurgia
3.
JA Clin Rep ; 6(1): 55, 2020 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-32681473

RESUMO

BACKGROUND: Chronic thromboembolic pulmonary hypertension (CTEPH) is a disease of obstructive pulmonary artery remodeling as a consequence of major vessel thromboembolism. Balloon pulmonary angioplasty (BPA) is an alternative treatment for patients with inoperable CTEPH. We report a case of CTEPH which improved following preoperative BPA intervention, allowing total hysterectomy to be performed. CASE PRESENTATION: A 48-year-old woman was transferred to our hospital to undergo total hysterectomy for endometrial cancer. She developed pulmonary embolism 7 months ago at another hospital, and a diagnosis of CTEPH was made based on multiple pulmonary emboli and pulmonary hypertension at our institute. Two BPA sessions for seven branches of the bilateral pulmonary arteries were conducted, resulting in a decrease of mean pulmonary artery pressure from 54 to 33 mmHg. Total hysterectomy was successfully performed under general anesthesia without any complications. CONCLUSIONS: BPA could be effective for reducing PH in patients with CTEPH undergoing noncardiac surgery.

4.
Sci Total Environ ; 708: 134659, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-31791760

RESUMO

Huge tsunami waves devastated coastal areas on the Pacific Ocean side of northern Japan on March 11, 2011, and seriously damaged these coastal environments. Since the tsunami, we have not yet obtained data on the present state of and changes in trace metal concentrations in seawater in these areas. Platinum (Pt), one of the rarest elements in the Earth's crust, is now widely used in a range of products, such as catalytic converters in automobiles and anticancer drugs. Increasing use and dispersal of Pt has the potential to affect aquatic environments, although Pt concentrations in open ocean seawater have been found to be very low (approximately 0.2 pmol/L). In this study, we reveal the Pt concentrations in seawater and sediment in Otsuchi Bay after the tsunami, and evaluate the behavior of the Pt. The concentrations of dissolved Pt in seawater are 0.40-1.99 pmol/L and those in river water are below the detection limit of 0.015 pmol/L. Comparing the Pt concentrations in May, higher concentrations were obtained in 2013 than in 2012, especially in the deepest seawater. The total Pt concentrations in sediment samples were 0.46-14.4 ng/g in Otsuchi Bay. Using a sequential leaching technique on the sediments, Pt concentrations in the acetic-acid fractions were 0.19-1.13 ng/g, and those in the acetic acid + hydrochloric acid hydroxylamine fractions were less than 0.03-0.71 ng/g. Seasonal variations in dissolved Pt concentrations reflected changes in the water mass structure. During the stratification season, vertical profiles indicated that Pt concentrations tended to increase with depth due to supply from the sediments, whereas in winter, the water mass was vertically well mixed. The Pt was supplied to the bottom of the water from the sediments, probably due to loosely adsorbed Pt on sediment particles being remobilized during post-depositional processes. The increased internal input of Pt within Otsuchi Bay can be explained by the release of 1.3-5.6% of the leachable fraction from sediments, probably transported from the land by the tsunami, during the water residence time in the bay.

5.
Environ Health Prev Med ; 24(1): 81, 2019 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-31883515

RESUMO

BACKGROUND: Human T cell leukemia virus type-1 (HTLV-1) stimulates inflammation activity. Our previous study revealed a positive association between asymptomatic HTLV-1 infection and advanced periodontitis among elderly Japanese individuals with low levels of hematopoietic activity (reflected by reticulocyte levels). Since low hematopoietic activity has been correlated with low-grade inflammation and low-grade inflammation is associated with atherosclerosis, the status of atherosclerosis could, in turn, determine the nature of this association. METHODS: To this end, a cross-sectional study of 907 elderly Japanese individuals (aged 60-99 years), who had participated in dental health check-up during the period 2016-2018, was conducted. Advanced periodontitis was defined as periodontal pocket ≥ 6.0 mm. RESULTS: Among the study population, 295 (32.5%) were found to have atherosclerosis defined as a carotid intima-media thickness (CIMT) of ≥ 1.1 mm. HTLV-1 infection was positively associated with advanced periodontitis in participants with atherosclerosis, but no significant associations were observed among the participants without atherosclerosis. The known risk factors' (including reticulocyte and CIMT) adjusted odds ratio (OR) and 95% confidence interval (CI) of advanced periodontitis were OR 2.01 and 95% CI 1.06-3.81 for participants with atherosclerosis and OR 0.61 and 95% CI 0.34-1.12 for participants without atherosclerosis. CONCLUSION: This study found a significant association between HTLV-1 infection and advanced periodontitis among elderly Japanese with atherosclerosis. However, this association is absent in individuals without atherosclerosis, suggesting that atherosclerosis might act as a determinant in the association between HTLV-1 infection and advanced periodontitis among elderly Japanese.


Assuntos
Aterosclerose/epidemiologia , Infecções por HTLV-I/epidemiologia , Vírus Linfotrópico T Tipo 1 Humano/fisiologia , Periodontite/epidemiologia , Idoso , Idoso de 80 Anos ou mais , Aterosclerose/etiologia , Espessura Intima-Media Carotídea , Estudos Transversais , Feminino , Infecções por HTLV-I/virologia , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Razão de Chances , Periodontite/etiologia , Fatores de Risco
6.
Environ Health Prev Med ; 24(1): 42, 2019 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-31182010

RESUMO

BACKGROUND: We reported that human T cell leukemia virus 1 (HTLV-1) infection is positively associated with atherosclerosis. Recent evidence has revealed a close association of periodontitis with atherosclerosis, endothelial dysfunction, and disruption of the microcirculation. However, the association between HTLV-1 and advanced periodontitis has not been investigated to date. Since hematopoietic activity is closely linked to endothelial maintenance activity and is known to decline with age, we hypothesized that the state of hematopoietic activity influenced the association between HTLV-1 and advanced periodontitis in elderly participants. METHODS: A cross-sectional study was performed including 822 elderly participants aged 60-99 years who participated in a dental health check-up. Advanced periodontitis was defined as a periodontal pocket ≥ 6.0 mm. Participants were classified as having low or high hematopoietic activity according to the median values of reticulocytes. RESULTS: HTLV-1 infection was positively related to advanced periodontitis among participants with lower hematopoietic activity (lower reticulocyte count), but not among participants with higher hematopoietic activity (higher reticulocyte count). The adjusted odds ratio (95% confidence interval) considering potential confounding factors was 1.92 (1.05-3.49) for participants with a lower reticulocyte count and 0.69 (0.35-1.36) for participants with a higher reticulocyte count. CONCLUSIONS: Among elderly participants, the association between HTLV-1 infection and advanced periodontitis is influenced by hematopoietic activity. Since hematopoietic activity is associated with endothelial maintenance, these findings provide an efficient tool for clarifying the underlying mechanism of the progression of periodontitis among elderly participants.


Assuntos
Infecções por HTLV-I/fisiopatologia , Hematopoese/fisiologia , Periodontite/epidemiologia , Idoso , Idoso de 80 Anos ou mais , Estudos Transversais , Feminino , Vírus Linfotrópico T Tipo 1 Humano/fisiologia , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Razão de Chances , Periodontite/virologia , Prevalência , Fatores de Risco
7.
Masui ; 61(10): 1088-90, 2012 Oct.
Artigo em Japonês | MEDLINE | ID: mdl-23157092

RESUMO

We gave anesthesia for tracheal separation in a patient with Creutzfeldt-Jakob disease. The patient, a 33-year-old woman, was bedridden and unable to communicate, and was going to undergo a tracheal separation procedure for repeated bouts of aspiration pneumonia. After a tracheostomy with local anesthesia and sedation with propofol, general anesthesia was induced and maintained with propofol (1.5-3.0 microg x ml(-1), target controlled infusion) and remifentanil (0.05-0.15 microg x kg(-1) x min(-1)). We did not use an anesthetic apparatus from the standpoint of infection control, and provided manual ventilation with a disposable Jackson-Rees circuit. During the operation, an entropy monitor indicated alternating extremely low (0-10) and high (90-100) values without circulatory change, probably due to a previously existing electroencephalographic abnormality. The surgery was uneventful, and spontaneous breathing and eyelid opening occurred about 10 minutes after discontinuation of remifentanil and propofol. In such infected patients, abnormal prion proteins can exist outside of the central nervous system throughout the period of anesthetic management. Therefore, careful infection control must be undertaken, even if the surgical site is not directly related to the central nervous system.


Assuntos
Anestesia Geral/métodos , Síndrome de Creutzfeldt-Jakob/cirurgia , Controle de Infecções/métodos , Traqueia/cirurgia , Adulto , Feminino , Humanos , Pneumonia Aspirativa/prevenção & controle , Propofol , Prevenção Secundária
8.
PLoS One ; 7(5): e37136, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22623985

RESUMO

G-protein-coupled receptors (GPCRs) regulate a wide variety of physiological processes and are important pharmaceutical targets for drug discovery. Here, we describe a unique concept based on yeast cell-surface display technology to selectively track eligible peptides with agonistic activity for human GPCRs (Cell Wall Trapping of Autocrine Peptides (CWTrAP) strategy). In our strategy, individual recombinant yeast cells are able to report autocrine-positive activity for human GPCRs by expressing a candidate peptide fused to an anchoring motif. Following expression and activation, yeast cells trap autocrine peptides onto their cell walls. Because captured peptides are incapable of diffusion, they have no impact on surrounding yeast cells that express the target human GPCR and non-signaling peptides. Therefore, individual yeast cells can assemble the autonomous signaling complex and allow single-cell screening of a yeast population. Our strategy may be applied to identify eligible peptides with agonistic activity for target human GPCRs.


Assuntos
Comunicação Autócrina/fisiologia , Bioensaio/métodos , Parede Celular/metabolismo , Peptídeos/isolamento & purificação , Peptídeos/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Biotecnologia/métodos , Western Blotting , Citometria de Fluxo , Imunofluorescência , Humanos , Feromônios Humano/metabolismo , Plasmídeos/genética , Receptores Acoplados a Proteínas G/agonistas , Saccharomyces cerevisiae
9.
Biotechnol J ; 7(5): 686-9, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22294378

RESUMO

Glutathione is a valuable tripeptide that is widely used in the pharmaceutical, food, and cosmetic industries. Glutathione is industrially produced by fermentation using Saccharomyces cerevisiae. Before the glutathione fermentation process with S. cerevisiae, a glucose extraction process from starchy materials is required. This glucose extraction is usually carried out by converting starchy materials to starch using high-temperature cooking and subsequent hydrolysis by amylases to convert starch to glucose. In this study, to develop an energy-saving glutathione production process by reducing energy consumption during the cooking step, we efficiently produced glutathione from low-temperature cooked rice using amylase-expressing S. cerevisiae. The combination of the amylase-expressing yeast with low-temperature cooking is potentially applicable to a variety of energy-saving bio-production methods of chemicals from starchy bio-resources.


Assuntos
Glucana 1,4-alfa-Glucosidase/metabolismo , Glutationa/metabolismo , Oryza/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Biomassa , Reatores Biológicos/microbiologia , Culinária , Glucose/metabolismo , Oryza/química , Tamanho da Partícula , Saccharomyces cerevisiae/enzimologia , Amido/metabolismo , Temperatura
10.
Biotechnol J ; 7(5): 642-8, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-21922670

RESUMO

Modification of proteins with small molecules is a widely used and powerful tool in biological research. Enzymatic approaches are particularly promising because substrate specificity allows for site-specific modification. Sortase A, a transpeptidase from Staphylococcus aureus, cleaves between the T and G residues in the sequence LPXTG, and subsequently links the carboxyl group of the T residue to an amino group of N-terminal glycine oligomers by a native peptide bond. Although Gram-positive bacteria have several kinds of sortases, there are few reports concerning their expression and substrate specificity. Here, we demonstrate site-specific protein modification with primary amine-containing molecules catalyzed by Lactobacillus plantarum sortase. Enhanced green fluorescent protein (EGFP) was employed as a model protein, and an amine-containing biotin molecule was site-specifically conjugated with LPQTSEQ-tagged EGFP. We developed a novel Lactobacillus plantarum sortase that has different substrate specificity compared to Staphylococcus aureus sortase. Amine-directed protein modification was achieved using the Lactobacillus plantarum sortase ''LPQTSEQ'' sequence original recognition tag. Our results demonstrate a promising method for expanding the capabilities of site-specific protein-small molecule modification.


Assuntos
Aminoaciltransferases/metabolismo , Proteínas de Bactérias/metabolismo , Biotecnologia/métodos , Cisteína Endopeptidases/metabolismo , Lactobacillus plantarum/enzimologia , Engenharia de Proteínas/métodos , Sequência de Aminoácidos , Aminoaciltransferases/química , Proteínas de Bactérias/química , Biotina/química , Biotina/metabolismo , Cisteína Endopeptidases/química , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/metabolismo , Indicadores e Reagentes/química , Indicadores e Reagentes/metabolismo , Dados de Sequência Molecular , Oligopeptídeos/química , Oligopeptídeos/metabolismo , Ligação Proteica
11.
Appl Microbiol Biotechnol ; 93(4): 1495-502, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22075633

RESUMO

Glutathione is a valuable tri-peptide that is widely used in the pharmaceutical, food, and cosmetic industries. Glutathione is produced industrially by fermentation using Saccharomyces cerevisiae, and supplementation of fermentation with several amino acids can increase intracellular GSH content. More recently, however, focus has been given to protein as a resource for biofuel and fine chemical production. We demonstrate that expression of a protease on the cell surface of S. cerevisiae enables the direct use of keratin and soy protein as a source of amino acids and that these substrates enhanced intracellular GSH content. Furthermore, fermentation using soy protein also enhanced cell concentration. GSH fermentation from keratin and to a greater extent from soy protein using protease-displaying yeast yielded greater GSH productivity compared to GSH fermentation with amino acid supplementation. This protease-displaying yeast is potentially applicable to a variety of processes for the bio-production of value-added chemicals from proteinaceous biomass resources.


Assuntos
Biomassa , Glutationa/isolamento & purificação , Glutationa/metabolismo , Peptídeo Hidrolases/metabolismo , Proteínas/metabolismo , Proteólise , Saccharomyces cerevisiae/enzimologia , Fermentação , Expressão Gênica , Queratinas/metabolismo , Engenharia Metabólica , Peptídeo Hidrolases/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Proteínas de Soja/metabolismo
12.
Enzyme Microb Technol ; 48(4-5): 393-6, 2011 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-22112955

RESUMO

Efficient ethanol producing yeast Saccharomyces cerevisiae cannot produce ethanol from raw starch directly. Thus the conventional ethanol production required expensive and complex process. In this study, we developed a direct and efficient ethanol production process from high-yielding rice harvested in Japan by using amylase expressing yeast without any pretreatment or addition of enzymes or nutrients. Ethanol productivity from high-yielding brown rice (1.1g/L/h) was about 5-fold higher than that obtained from purified raw corn starch (0.2g/L/h) when nutrients were added. Using an inoculum volume equivalent to 10% of the fermentation volume without any nutrient supplementation resulted in ethanol productivity and yield reaching 1.2g/L/h and 101%, respectively, in a 24-h period. High-yielding rice was demonstrated to be a suitable feedstock for bioethanol production. In addition, our polyploid amylase-expressing yeast was sufficiently robust to produce ethanol efficiently from real biomass. This is first report of direct ethanol production on real biomass using an amylase-expressing yeast strain without any pretreatment or commercial enzyme addition.


Assuntos
Amilases/metabolismo , Biotecnologia/métodos , Etanol/metabolismo , Oryza/metabolismo , Saccharomyces cerevisiae/enzimologia , Biomassa , Meios de Cultura , Fermentação , Japão , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/metabolismo , alfa-Amilases/metabolismo
13.
Appl Microbiol Biotechnol ; 91(4): 1001-6, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21573687

RESUMO

We developed a novel enzymatic glutathione (GSH) production system using Saccharomyces cerevisiae as a whole-cell biocatalyst, and improved its GSH productivity by metabolic engineering. We demonstrated that the metabolic engineering of GSH pathway and ATP regeneration can significantly improve GSH productivity by up to 1.7-fold higher compared with the parental strain, respectively. Furthermore, the combination of both improvements in GSH pathway and ATP regeneration is more effective (2.6-fold) than either improvement individually for GSH enzymatic production using yeast. The improved whole-cell biocatalyst indicates its great potential for applications to other kinds of ATP-dependent bioproduction.


Assuntos
Enzimas/metabolismo , Engenharia Genética , Glutationa/metabolismo , Redes e Vias Metabólicas/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Trifosfato de Adenosina/metabolismo
14.
Masui ; 60(4): 470-2, 2011 Apr.
Artigo em Japonês | MEDLINE | ID: mdl-21520598

RESUMO

Porphyria is a hereditary disorder due to reduction in enzyme activity of heme synthesis system, resulting in accumulation of heme precursors. Erythropoietic protoporphyria (EPP) is a type of porphyria. There are varieties of clinical expressions of EPP such as hepatic damage and photosensitivity, caused by the accumulation of protoporphyrin in the liver and the skin. Therefore it is important to prevent development of these clinical expressions. A 36-year-old woman with EPP was scheduled for a laparoscopic appendectomy. Anesthesia was induced with remifentanil, sevoflurane and rocuronium, and maintained with remifentanil, fentanyl and sevoflurane. She had performed normal daily activities without taking countermeasures against photosensitivity, so we did not change the light in the operating room, and allowed using endoscopy. The surgery was performed without any complications. No skin symptom was observed perioperatively. The level of aminotransferase was elevated temporally after the operation, while the level of protoporphyrin was unchanged. Therefore, we considered there was no manifestation of EPP perioperatively. In general, patients with EPP do not develop acute attacks induced by drugs like barbiturates. Since there was a case report of severe liver dysfunction of EPP, we did not use any contraindicated drugs in patients with acute intermittent porphyria in this patient. Considering a possibility of motor nerve damage in severe EPP, we carried out anesthetic management without epidural anesthesia.


Assuntos
Anestesia Geral/métodos , Apendicectomia , Laparoscopia , Protoporfiria Eritropoética/complicações , Adulto , Feminino , Humanos
15.
J Biotechnol ; 152(1-2): 37-42, 2011 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-21262280

RESUMO

Streptavidin is tetrameric protein which has tight and specific biotin binding affinity, and streptavidin modification of proteins or small molecules is widely used for biotechnology tool. Here, we demonstrate site-specific streptavidin-protein conjugation using enzymes. We focused on sortase A, a transpeptidase from Staphylococcus aureus. A streptavidin-tagged LPETG motif (Stav-LPETG) was expressed in Escherichia coli. We achieved soluble streptavidin expression in E. coli without refolding using a cold shock expression system. Then we successfully conjugated Stav-LPETG with pentaglycine-appended green fluorescence protein (Gly5-GFP) or triglycine-appended glucose oxidase (Gly3-GOD) using sortase A. SDS-PAGE analysis showed site-specific tetrameric streptavidin-protein conjugation with the tagged proteins. In addition, the functions of a Stav-GOD conjugate, i.e., biotin-binding and glucose oxidase activity, were significantly higher compared to those of streptavidin-GOD conjugates prepared by chemical modification.


Assuntos
Aminoaciltransferases/metabolismo , Proteínas de Bactérias/metabolismo , Cisteína Endopeptidases/metabolismo , Estreptavidina/metabolismo , Aminoaciltransferases/genética , Proteínas de Bactérias/genética , Cisteína Endopeptidases/genética , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Escherichia coli/metabolismo , Glucose Oxidase/genética , Glucose Oxidase/metabolismo , Estreptavidina/genética
16.
Appl Microbiol Biotechnol ; 89(5): 1417-22, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21104244

RESUMO

Glutathione is a valuable tri-peptide that is widely used in the pharmaceutical, food, and cosmetic industries. Glutathione is produced industrially by fermentation using Saccharomyces cerevisiae. We demonstrated that expression of amylase genes in glutathione-producing S. cerevisiae enables direct use of starch as a carbon source, thus eliminating the Crabtree effect that is caused by excess glucose. Consequently, cell growth and glutathione productivity were significantly improved. This approach is potentially applicable to a variety of fermentative processes for production of value-added chemicals under aerobic conditions.


Assuntos
Glutationa/biossíntese , Saccharomyces cerevisiae/metabolismo , Amido/metabolismo , Amilases/genética , Amilases/metabolismo , Redes e Vias Metabólicas/genética , Organismos Geneticamente Modificados , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crescimento & desenvolvimento , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo
17.
Bioconjug Chem ; 21(12): 2227-33, 2010 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-21069999

RESUMO

A ZZ domain (ZZ) and alkaline phosphatase (AP), luciferase (Luc), or glucose oxidase (GOD) were conjugated using Sortase A (SrtA) from Staphylococcus aureus. The specific peptidyl linker for SrtA was genetically fused to the C-terminus of ZZ, and the other linker was fused to the N-terminus of AP, Luc, or GOD, respectively. The resultant proteins were obtained separately by bacterial expression. The recombinant peptide-tagged ZZ and AP, Luc, or GOD were site-specifically conjugated by SrtA through the extra peptidyl linkers in vitro. The SrtA reaction had little influence on either the antibody-binding activity of the ZZ moiety or the enzymatic activity of AP, Luc, or GOD moieties of the conjugates. Additionally, antibody-ZZ-proteins were yielded easily by mixing antibody with ZZ-AP, ZZ-Luc, or ZZ-GOD, allowing their use in an enzyme-linked immunosorbent assay. These results suggest that the enzymatic approach with SrtA facilitates the construction of ZZ-proteins. Furthermore, mixing antibody and ZZ-proteins produces a wide variety of antibody-ZZ-proteins.


Assuntos
Aminoaciltransferases , Proteínas de Bactérias/metabolismo , Cisteína Endopeptidases , Proteínas Fúngicas/metabolismo , Imunoconjugados/metabolismo , Engenharia de Proteínas/métodos , Proteínas Recombinantes de Fusão/metabolismo , Fosfatase Alcalina/análise , Fosfatase Alcalina/metabolismo , Aminoaciltransferases/genética , Aminoaciltransferases/metabolismo , Anticorpos/genética , Anticorpos/imunologia , Anticorpos/metabolismo , Antígenos/genética , Antígenos/imunologia , Antígenos/metabolismo , Aspergillus oryzae , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Cisteína Endopeptidases/genética , Cisteína Endopeptidases/metabolismo , Ensaio de Imunoadsorção Enzimática/métodos , Escherichia coli , Proteínas Fúngicas/genética , Proteínas Fúngicas/imunologia , Expressão Gênica , Glucose Oxidase/análise , Glucose Oxidase/metabolismo , Imunoconjugados/genética , Imunoconjugados/imunologia , Luciferases/análise , Luciferases/metabolismo , Ligação Proteica/genética , Ligação Proteica/imunologia , Estrutura Terciária de Proteína/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Staphylococcus aureus
18.
Bioorg Med Chem Lett ; 20(19): 5726-31, 2010 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-20801029

RESUMO

A novel HER2-targeted carrier was developed using bionanocapsules (BNCs). Bionanocapsules (BNCs) are 100-nm hollow nanoparticles composed of the L-protein of hepatitis B virus surface antigen. An affibody of HER2 was genetically displayed on the BNC surface (Z(HER2)-BNC). For the investigation of binding affinity, Z(HER2)-BNC was incubated with the cancer cell lines SK-BR-3 (HER2 positive), and MDA-MB-231 (HER2 negative). For analysis of HER2 targeting specificity, Z(HER2)-BNC or Z(WT)-BNC (without affibody) was incubated with both SK-BR-3 and MDA-MB-231 cells by time lapse and concentration. For the delivery of encapsulated molecules (calcein), fluorescence of Z(HER2)-BNC mixed with liposomes was also compared with that of Z(WT)-BNC and nude liposomes by incubation with SK-BR-3 cells. As a result, Z(HER2)-BNC-liposome complex demonstrated the delivery to HER2-expressing cells (SK-BR-3) with a high degree of specificity. This indicates that genetically engineered BNCs are promising carrier for cancer treatment.


Assuntos
Nanopartículas/administração & dosagem , Receptor ErbB-2/metabolismo , Proteínas Recombinantes de Fusão/administração & dosagem , Neoplasias da Mama/tratamento farmacológico , Linhagem Celular Tumoral , Portadores de Fármacos/química , Feminino , Fluoresceínas/química , Antígenos da Hepatite B/genética , Antígenos da Hepatite B/metabolismo , Humanos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo
19.
Appl Microbiol Biotechnol ; 85(5): 1491-8, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19707752

RESUMO

We developed a novel strategy for constructing yeast to improve levels of amylase gene expression and the practical potential of yeast by combining delta-integration and polyploidization through cell fusion. Streptococcus bovis alpha-amylase and Rhizopus oryzae glucoamylase/alpha-agglutinin fusion protein genes were integrated into haploid yeast strains. Diploid strains were constructed from these haploid strains by mating, and then a tetraploid strain was constructed by cell fusion. The alpha-amylase and glucoamylase activities of the tetraploid strain were increased up to 1.5- and tenfold, respectively, compared with the parental strain. The diploid and tetraploid strains proliferated faster, yielded more cells, and fermented glucose more effectively than the haploid strain. Ethanol productivity from raw starch was improved with increased ploidy; the tetraploid strain consumed 150 g/l of raw starch and produced 70 g/l of ethanol after 72 h of fermentation. Our strategy for constructing yeasts resulted in the simultaneous overexpression of genes integrated into the genome and improvements in the practical potential of yeasts.


Assuntos
Etanol/metabolismo , Engenharia Genética , Glucana 1,4-alfa-Glucosidase/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Amido/metabolismo , alfa-Amilases/genética , Aerobiose , Anaerobiose , Clonagem Molecular , Fermentação , Regulação Fúngica da Expressão Gênica , Glucana 1,4-alfa-Glucosidase/metabolismo , Poliploidia , Proteínas Recombinantes/metabolismo , Rhizopus/enzimologia , Rhizopus/genética , Saccharomyces cerevisiae/crescimento & desenvolvimento , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Streptococcus bovis/enzimologia , Streptococcus bovis/genética , Especificidade por Substrato , Transformação Genética , alfa-Amilases/metabolismo
20.
Masui ; 58(10): 1278-81, 2009 Oct.
Artigo em Japonês | MEDLINE | ID: mdl-19860233

RESUMO

Hunter syndrome, manifested by mucopolysaccharidosis II (MPS II), is a hereditary disorder caused by accumulation of glycosaminoglycans. An important issue in regard to anesthesia in affected individuals is airway management, because of gargoylism. An 8-year-old boy with MPS II was scheduled for adenotonsillectomy. We performed slow induction with sevoflurane, then utilized an Airway Scope (AWS; HOYA Corporation Tokyo, Japan), which enables operators to observe tube passage through the vocal cords with an LCD monitor, for tracheal intubation, because intubation guided by a fiberoptic bronchoscopy (FOB) was considered to be risky for glottic damage. Attempted tracheal intubations with the AWS alone and in combination with an FOB inserted through the tracheal tube failed, even though a clear image of the glottis was obtained. Finally, we inserted a stylet into the tracheal tube attached to the blade of the AWS and successful tracheal intubation was accomplished. The operation was completed uneventfully and the patient entered the ICU with his trachea intubated, because of pharyngeal and laryngeal edema. Although useful for difficult airway management, tracheal intubation with the AWS may be difficult when used in patients with a narrow oral cavity or small tracheal tube.


Assuntos
Anestesia Geral , Intubação Intratraqueal/efeitos adversos , Edema Laríngeo/etiologia , Mucopolissacaridose II/cirurgia , Adenoidectomia , Criança , Humanos , Intubação Intratraqueal/instrumentação , Intubação Intratraqueal/métodos , Masculino , Tonsilectomia
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