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1.
BMC Oral Health ; 24(1): 24, 2024 01 05.
Artigo em Inglês | MEDLINE | ID: mdl-38183059

RESUMO

BACKGROUND: The profound influence of orthodontic treatments on facial aesthetics has been a topic of increasing interest. This study delves into the intricate interplay between orthodontic treatments, facial feature alterations, and aesthetic perceptions. METHODS: A total of 73 patients who had undergone orthodontic treatment were included in this study. Facial photographs were taken before and after treatment. Ten orthodontists provided facial aesthetic ratings (FAR) for each patient's frontal, profile, and overall views. 48 facial landmarks were manually placed by the orthodontists and normalized using Generalized Procrustes analysis (GPA). Two types of phenotypes were derived from facial landmarks. Global facial phenotypes were then extracted using principal component analysis (PCA). Additionally, 37 clinical features related to aesthetics and orthodontics were extracted. The association between facial features and changes in FAR after orthodontic treatment was determined using these two types of phenotypes. RESULTS: The FAR exhibited a high correlation among orthodontic experts, particularly in the profile view. The FAR increased after orthodontic treatment, especially in profile views. Extraction of premolars and orthognathic surgery were found to result in higher FAR change. For global facial phenotypes, the most noticeable changes in the frontal and profile views associated with FAR occurred in the lip area, characterized by inward retraction of the lips and slight chin protrusion in the profile view, as well as a decrease in lip height in the frontal view. The changes observed in the profile view were statistically more significant than those in the frontal view. These facial changes were consistent with the changes from orthodontic treatment. For clinical features, two profile features, namely pg.sm.hori and pg.n.ls, were found to be associated with FAR following orthodontic treatment. The highest FAR scores were achieved when pg.sm.hori was at 80° and pg.n.ls was at 8°. On the other hand, frontal clinical features had a subtle effect on FAR during orthodontic treatment. CONCLUSIONS: This study demonstrated that orthodontic treatment improves facial aesthetics, particularly at lip aera in the profile view. Profile clinical features, such as pg.sm.hori and pg.n.ls, are essential in orthodontic treatment which could increase facial aesthetics.


Assuntos
Estética Dentária , Face , Humanos , Estudos Retrospectivos , Lábio , Queixo
2.
J Econ Entomol ; 114(2): 937-946, 2021 04 13.
Artigo em Inglês | MEDLINE | ID: mdl-33459777

RESUMO

Microbiome associated with insects play vital roles in host ecology and physiology. The small brown planthopper (SBPH), Laodelphax striatellus, is a polyphagous insect pest that caused enormous damage to a wide range of cereal crops. Previous studies have assessed the effects of environmental factors, such as antibiotics, insecticide, and geographical habitat on the bacterial composition of SBPH. However, the influence of host plants on the microbial community in SBPH still unclear. Here, we characterized and compared the microbial community in three SBPH populations feeding on rice, barley, and wheat, respectively, using high-throughput amplicon sequencing. Our observations revealed that the microbiome harbored by SBPH was abundant and diverse. Ten phyla comprising 141 genera of bacteria were annotated, and four fungal phyla consisting of 47 genera were assigned. The bacteria belonging to the phylum Proteobacteria were the most prevalent and the fungi with the highest abundance were from the order Hypocreales. Comparative analysis showed that host plants could significantly induce structural changes of SBPH microbiome. Significant differences in abundance were observed in two main bacterial orders (Rickettsiales and Rhodospirillales) and three fungal classes (Sordariomycetes, an unclassified class in Ascomycota and Eurotiomycetes) among three host-adapted SBPH populations. Our results could broaden our understanding of interactions among SBPH, its microbial associates and host plants, and also represented the basis of future SBPH biological management.


Assuntos
Hemípteros , Microbiota , Oryza , Animais , Bactérias/genética , Fungos
3.
Kaohsiung J Med Sci ; 37(2): 121-127, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33022894

RESUMO

Cullin 4B (CUL4B) was reported to be closely related to the progression of some tumors, but its function in clear cell renal cell carcinoma (ccRCC) has not been reported. Our present study found CUL4B was upregulated in ccRCC, and CUL4B knockdown markedly inhibited ccRCC cell growth and induced apoptosis. In addition, CUL4B knockdown markedly inhibited antiapoptotic proteins' expression in ccRCC cells, including Mcl-1 and Bcl-2, and silenced CUL4B also induced the cleavages of PARP, an important index of apoptosis. We also confirmed microRNA-217 (miR-217) was downregulated in ccRCC tumor tissues, and negatively correlated with CUL4B expression. Further investigations revealed miR-217 targeted CUL4B and markedly inhibited its expression in ccRCC cells. In addition, overexpression of miR-217 by mimics significantly suppressed ccRCC cell growth. In contrast, enforced expression of CUL4B significantly abolished miR-217-induced cell survival inhibition in ccRCC cells. In conclusion, our present results suggested targeting miR-217-CUL4B axis would be a promising strategy for ccRCC treatment.


Assuntos
Apoptose , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/patologia , Proteínas Culina/metabolismo , Neoplasias Renais/genética , Neoplasias Renais/patologia , MicroRNAs/metabolismo , Apoptose/genética , Sequência de Bases , Linhagem Celular Tumoral , Proliferação de Células/genética , Sobrevivência Celular/genética , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , MicroRNAs/genética
4.
Rapid Commun Mass Spectrom ; 33(9): 875-882, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30821059

RESUMO

RATIONALE: Heteroatomic compounds are relatively abundant and believed to be bio-resistant in heavy crude oils. However, few studies have focused on the biodegradation of these heteroatomic compounds. METHODS: Heteroatoms, especially N1 species, in a blank crude oil and in three treated oils co-incubated with anaerobic sulfate-reducing bacteria, nitrate-reducing bacteria and fermentative consortia cultures were detected using negative-ion electrospray ionization coupled with high-field Fourier transform ion cyclotron resonance mass spectrometry. RESULTS: The relative abundance of N1 species in the three treated oils decreased, while the relative abundance of O2 species increased. Remarkably, the relative abundances of N1 species with low carbon number increased and those with higher carbon number decreased. CONCLUSIONS: These results revealed that the anaerobic biodegradations of heavy crude oil occurred. With direct evidences, the degradations of alkyl side chains of N1 species by the anaerobic microbes could be deduced.

5.
Molecules ; 21(10)2016 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-27681713

RESUMO

Phytochemical investigation of the twigs of Podocarpus nagi (Podocarpaceae) led to the isolation of two new abietane-type diterpenoids, named 1ß,16-dihydroxylambertic acid (1) and 3ß,16-dihydroxylambertic acid (2), along with two new ent-pimarane-type diterpenoids, named ent-2ß,15,16,18-tetrahydroxypimar-8(14)-ene (3) and ent-15-oxo-2ß,16,18-trihydroxypimar-8(14)-ene (4). Their respective structures were elucidated on the basis of spectroscopic analyses, including 1D- and 2D-NMR, IR, CD, and HR-ESI-MS. This is the first time ent-pimarane-type diterpenoids from the genus Podocarpus has been reported. All four new compounds were tested for cytotoxic activity. The MTT assay results showed that compounds 3 and 4 significantly inhibited the proliferation of human cervical cancer Hela cells, human lung cancer A549 cells, and human breast cancer MCF-7 cells at a concentration of 10 µM. Furthermore, using the lipopolysaccharide (LPS)-stimulated RAW264.7 cells, compounds 2 and 4 were found to significantly inhibit nitrogen oxide (NO) production with IC50 values of 26.5 ± 6.1 and 17.1 ± 1.5 µM, respectively.

6.
Gene ; 590(2): 298-306, 2016 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-27259661

RESUMO

The complete mitogenomes of two orb-weaving spiders Neoscona doenitzi and Neoscona nautica were determined and a comparative mitogenomic analysis was performed to depict evolutionary trends of spider mitogenomes. The circular mitogenomes are 14,161bp with A+T content of 74.6% in N. doenitzi and 14,049bp with A+T content of 78.8% in N. nautica, respectively. Both mitogenomes contain a standard set of 37 genes typically presented in metazoans. Gene content and orientation are identical to all previously sequenced spider mitogenomes, while gene order is rearranged by tRNAs translocation when compared with the putative ancestral gene arrangement pattern presented by Limulus polyphemus. A comparative mitogenomic analysis reveals that the nucleotide composition bias is obviously divergent between spiders in suborder Opisthothelae and Mesothelae. The loss of D-arm in the trnS(UCN) among all of Opisthothelae spiders highly suggested that this common feature is a synapomorphy for entire suborder Opisthothelae. Moreover, the trnS(AGN) in araneoids preferred to use TCT as an anticodon rather than the typical anticodon GCT. Phylogenetic analysis based on the 13 protein-coding gene sequences consistently yields trees that nest the two Neoscona spiders within Araneidae and recover superfamily Araneoidea as a monophyletic group. The molecular information acquired from the results of this study should be very useful for future research on mitogenomic evolution and genetic diversities in spiders.


Assuntos
Genoma Mitocondrial , Filogenia , Aranhas/genética , Animais , Composição de Bases/genética , Sequência de Bases , Códon/genética , Genes Mitocondriais , Conformação de Ácido Nucleico , Nucleotídeos/genética , Fases de Leitura Aberta/genética , RNA Ribossômico/genética , RNA de Transferência/genética
7.
J Zhejiang Univ Sci B ; 16(10): 865-74, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26465134

RESUMO

In this study, we isolated an environmental clone of Ochrobactrum intermedium, strain 2745-2, from the formation water of Changqing oilfield in Shanxi, China, which can degrade crude oil. Strain 2745-2 is aerobic and rod-shaped with optimum growth at 42 °C and pH 5.5. We sequenced the genome and found a single chromosome of 4 800 175 bp, with a G+C content of 57.63%. Sixty RNAs and 4737 protein-coding genes were identified: many of the genes are responsible for the degradation, emulsification, and metabolizing of crude oil. A comparative genomic analysis with related clinical strains (M86, 229E, and LMG3301(T)) showed that genes involved in virulence, disease, defense, phages, prophages, transposable elements, plasmids, and antibiotic resistance are also present in strain 2745-2.


Assuntos
Proteínas de Bactérias/genética , Ochrobactrum/genética , Ochrobactrum/isolamento & purificação , Petróleo/microbiologia , Microbiologia da Água , Ochrobactrum/classificação , Especificidade da Espécie
8.
Stand Genomic Sci ; 10: 9, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-27408680

RESUMO

Enterobacter mori strain 5-4 is a Gram-negative, motile, rod shaped, and facultatively anaerobic bacterium, which was isolated from a mixture of formation water (also known as oil-reservior water) and crude-oil in Karamay oilfield, China. To date, there is only one E. mori genome has been sequenced and very little knowledge about the mechanism of E. mori adapted to the petroleum reservoir. Here, we report the second E. mori genome sequence and annotation, together with the description of features for this organism. The 4,621,281 bp assembly genome exhibits a G + C content of 56.24% and contains 4,317 protein-coding and 65 RNA genes, including 5 rRNA genes.

9.
Mar Genomics ; 18 Pt B: 135-7, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25301038

RESUMO

Previous studies suggest that antibiotic resistance genes have an ancient origin, which is not always linked to the use of antibiotics but can be enhanced by human activities. Bacillus flexus strain T6186-2 was isolated from the formation water sample of a deep-subsurface oil reservoir. Interestingly, antimicrobial susceptibility testing showed that this strain is susceptible to kanamycin, however, resistant to ampicillin, erythromycin, gentamicin, vancomycin, fosfomycin, fosmidomycin, tetracycline and teicoplanin. To explore our knowledge about the origins of antibiotic resistance genes (ARGs) in the relatively pristine environment, we sequenced the genome of B. flexus strain T6186-2 as a permanent draft. It represents the evidence for the existence of a reservoir of ARGs in nature among microbial populations from deep-subsurface oil reservoirs.


Assuntos
Bacillus/genética , Farmacorresistência Bacteriana Múltipla/genética , Genoma Bacteriano/genética , Água Subterrânea/microbiologia , Campos de Petróleo e Gás/microbiologia , Composição de Bases , Sequência de Bases , Primers do DNA/genética , Dados de Sequência Molecular , Análise de Sequência de DNA
10.
Microbiologyopen ; 3(4): 446-56, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24890829

RESUMO

Oil reservoirs are specific habitats for the survival and growth of microorganisms in general. Pseudomonas stutzeri which is believed to be an exogenous organism inoculated into oil reservoirs during the process of oil production was detected frequently in samples from oil reservoirs. Very little is known, however, about the distribution and genetic structure of P. stutzeri in the special environment of oil reservoirs. In this study, we collected 59 P. stutzeri 16S rRNA gene sequences that were identified in 42 samples from 25 different oil reservoirs and we isolated 11 cultured strains from two representative oil reservoirs aiming to analyze the diversity and genomovar assignment of the species in oil reservoirs. High diversity of P. stutzeri was observed, which was exemplified in the detection of sequences assigned to four known genomovars 1, 2, 3, 20 and eight unknown genomic groups of P. stutzeri. The frequent detection and predominance of strains belonging to genomovar 1 in most of the oil reservoirs under study indicated an association of genomovars of P. stutzeri with the oil field environments.


Assuntos
Variação Genética , Campos de Petróleo e Gás/microbiologia , Pseudomonas stutzeri/classificação , Pseudomonas stutzeri/genética , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Dados de Sequência Molecular , Filogenia , Pseudomonas stutzeri/isolamento & purificação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
11.
Sci Rep ; 2: 760, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23094135

RESUMO

Water-flooded oil reservoirs have specific ecological environments due to continual water injection and oil production and water recycling. Using 16S rRNA gene clone library analysis, the microbial communities present in injected waters and produced waters from four typical water-flooded oil reservoirs with different in situ temperatures of 25 °C, 40 °C, 55 °C and 70 °C were examined. The results obtained showed that the higher the in situ temperatures of the oil reservoirs is, the less the effects of microorganisms in the injected waters on microbial community compositions in the produced waters is. In addition, microbes inhabiting in the produced waters of the four water-flooded oil reservoirs were varied but all dominated by Proteobacteria. Moreover, most of the detected microbes were not identified as indigenous. The objective of this study was to expand the pictures of the microbial ecosystem of water-flooded oil reservoirs.


Assuntos
Óleos Combustíveis/microbiologia , Microbiologia da Água , Biodiversidade , China , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Ecossistema , Óleos Combustíveis/toxicidade , Filogenia , Proteobactérias/classificação , Proteobactérias/genética , Proteobactérias/isolamento & purificação , Temperatura , Poluentes Químicos da Água/toxicidade
12.
Sci Rep ; 2: 483, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22761991

RESUMO

The ascomycete fungus Beauveria bassiana is a pathogen of hundreds of insect species and is commercially produced as an environmentally friendly mycoinsecticide. We sequenced the genome of B. bassiana and a phylogenomic analysis confirmed that ascomycete entomopathogenicity is polyphyletic, but also revealed convergent evolution to insect pathogenicity. We also found many species-specific virulence genes and gene family expansions and contractions that correlate with host ranges and pathogenic strategies. These include B. bassiana having many more bacterial-like toxins (suggesting an unsuspected potential for oral toxicity) and effector-type proteins. The genome also revealed that B. bassiana resembles the closely related Cordyceps militaris in being heterothallic, although its sexual stage is rarely observed. A high throughput RNA-seq transcriptomic analysis revealed that B. bassiana could sense and adapt to different environmental niches by activating well-defined gene sets. The information from this study will facilitate further development of B. bassiana as a cost-effective mycoinsecticide.


Assuntos
Beauveria/genética , Evolução Biológica , Genoma Fúngico , Adaptação Biológica/genética , Animais , Beauveria/crescimento & desenvolvimento , Beauveria/metabolismo , Análise por Conglomerados , Cordyceps/genética , Cordyceps/crescimento & desenvolvimento , Cordyceps/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Ordem dos Genes , Genes Fúngicos , Genômica , Insetos/microbiologia , Filogenia , Transdução de Sinais , Transcriptoma
13.
Environ Microbiol ; 14(8): 2139-50, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22118579

RESUMO

Knockout and complement mutants of mannitol-1-phosphate dehydrogenase (MPD) and mannitol dehydrogenase (MTD) were constructed to probe the roles of both enzymes in the mannitol metabolism and multi-stress tolerances of entomopathogenic fungus Beauveria bassiana. Compared with wild-type and complement mutants, ΔBbMPD lost 99.5% MPD activity for reducing fructose-6-phosphate to mannitol-1-phosphate while ΔBbMTD lost 78.9% MTD activity for oxidizing mannitol to fructose. Consequently, mannitol contents in mycelia and conidia decreased 68% and 83% for ΔBbMPD, and 16% and 38% for ΔBbMTD, accompanied by greatly enhanced trehalose accumulations due to 81-87% decrease in their neutral trehalase expression. Mannitol as mere carbon source in a nitrate-based minimal medium suppressed the colony growth of ΔBbMTD instead of ΔBbMPD, and delayed more conidial germination of ΔBbMTD than ΔBbMPD. Based on median lethal responses, conidial tolerances to H(2) O(2) oxidation, UV-B irradiation and heat stress at 45°C decreased 38%, 39% and 22% in ΔBbMPD, and 18%, 16% and 11% in ΔBbMTD respectively. Moreover, ΔBbMPD and ΔBbMTD lost 14% and 7% of their virulence against Spodoptera litura larvae respectively. Our findings highlight the primary roles of MPD and MTD in mannitol metabolism and their significant contributions to multi-stress tolerances and virulence influential on the biocontrol potential of B.bassiana.


Assuntos
Beauveria/enzimologia , Manitol Desidrogenases/metabolismo , Manitol/metabolismo , Estresse Fisiológico/fisiologia , Desidrogenase do Álcool de Açúcar/metabolismo , Animais , Beauveria/efeitos dos fármacos , Beauveria/patogenicidade , Beauveria/efeitos da radiação , Perfilação da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Teste de Complementação Genética , Peróxido de Hidrogênio/farmacologia , Manitol Desidrogenases/genética , Mutação , Micélio/metabolismo , Oxidantes/farmacologia , Spodoptera/microbiologia , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/metabolismo , Esporos Fúngicos/efeitos da radiação , Desidrogenase do Álcool de Açúcar/genética , Raios Ultravioleta , Virulência/genética
14.
Appl Microbiol Biotechnol ; 95(3): 811-21, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22159733

RESUMO

Based on preliminary investigation of microbial populations in a high pour-point oil reservoir, an indigenous microbial enhanced oil recovery (MEOR) field trial was carried out. The purpose of the study is to reveal the impact of the indigenous MEOR process on microbial community structure in the oil reservoir using 16Sr DNA clone library technique. The detailed monitoring results showed significant response of microbial communities during the field trial and large discrepancies of stimulated microorganisms in the laboratory and in the natural oil reservoir. More specifically, after nutrients injection, the original dominant populations of Petrobacter and Alishewanella in the production wells almost disappeared. The expected desirable population of Pseudomonas aeruginosa, determined by enrichment experiments in laboratory, was stimulated successfully in two wells of the five monitored wells. Unexpectedly, another potential population of Pseudomonas pseudoalcaligenes which were not detected in the enrichment culture in laboratory was stimulated in the other three monitored production wells. In this study, monitoring of microbial community displayed a comprehensive alteration of microbial populations during the field trial to remedy the deficiency of culture-dependent monitoring methods. The results would help to develop and apply more MEOR processes.


Assuntos
Biota , Campos de Petróleo e Gás/microbiologia , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
15.
Appl Biochem Biotechnol ; 163(2): 223-34, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20652442

RESUMO

Three biosurfactant-producing indigenous microorganisms (XDS1, XDS2, XDS3) were isolated from a petroleum reservoir in the Daqing Oilfield (China) after polymer flooding. Their metabolic, biochemical, and oil-degradation characteristics, as well as their oil displacement in the core were studied. These indigenous microorganisms were identified as short rod bacillus bacteria with white color, round shape, a protruding structure, and a rough surface. Strains have peritrichous flagella, are able to produce endospores, are sporangia, and are clearly swollen and terminal. Bacterial cultures show that the oil-spreading values of the fermentation fluid containing all three strains are more than 4.5 cm (diameter) with an approximate 25 mN/m surface tension. The hydrocarbon degradation rates of each of the three strains exceeded 50%, with the highest achieving 84%. Several oil recovery agents were produced following degradation. At the same time, the heavy components of crude oil were degraded into light components, and their flow characteristics were also improved. The surface tension and viscosity of the crude oil decreased after being treated by the three strains of microorganisms. The core-flooding tests showed that the incremental oil recoveries were 4.89-6.96%. Thus, XDS123 treatment may represent a viable method for microbial-enhanced oil recovery.


Assuntos
Bacillus/metabolismo , Petróleo/microbiologia , Tensoativos/isolamento & purificação , Bacillus/classificação , Bacillus/isolamento & purificação , Biotecnologia , China , Fermentação , Flagelos/fisiologia , Hidrocarbonetos/metabolismo , Polímeros/química , Esporângios/fisiologia , Tensão Superficial , Tensoativos/química , Tensoativos/metabolismo , Viscosidade
16.
Zhonghua Yi Xue Za Zhi ; 90(44): 3131-4, 2010 Nov 30.
Artigo em Chinês | MEDLINE | ID: mdl-21211344

RESUMO

OBJECTIVE: To compare the safety and efficacy of Doppler-guided hemorrhoidal artery ligation and operative procedures for prolapse and hemorrhoids. METHODS: Ninety-two cases with a diagnosis of hemorrhoids from June 1, 2006 to June 1, 2008 at our hospital were collected and divided into the observation group (A, n = 40) and the control group (B, n = 52). They received Doppler-guided hemorrhoidal artery ligation and operative procedures for prolapse and hemorrhoids respectively. The efficacy of two groups was compared with regards to efficacy, operative duration, postoperative pain score, postoperative analgesic use, postoperative complications, hospitalization duration, recovery time, total treatment expenditure, patient satisfaction and 1-year follow-up. RESULTS: After treatment, two groups demonstrated similar profiles of efficacy, operative duration, patient satisfaction and the relapsing rate at 1 year. Group A was obviously better than Group B. with regards to recovery time, hospitalization duration and total treatment expenditure. Furthermore the incidence of such complications as postoperative pain, urinary retention, anal bulge, hemorrhage, perianal infections, anal edema and anal fissure was lower. CONCLUSION: Doppler-guided hemorrhoidal artery ligation has many advantages, such as safety, effectiveness, less trauma, a quick recovery, less treatment expenditure and a lower incidence of complications. Thus it is worthy of a wider popularization. However, its long-term effect remains to be seen.


Assuntos
Artérias/cirurgia , Hemorroidas/cirurgia , Ligadura/métodos , Adulto , Artérias/diagnóstico por imagem , Feminino , Hemorroidas/diagnóstico por imagem , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Reto/irrigação sanguínea , Resultado do Tratamento , Ultrassonografia , Procedimentos Cirúrgicos Vasculares
17.
Carbohydr Res ; 345(1): 50-4, 2010 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-19896118

RESUMO

A long-chain mannitol-1-phosphate dehydrogenase (MPD) was characterized for the first time from fungal entomopathogen Beauveria bassiana by gene cloning, heterogeneous expression and activity analysis. The cloned gene BbMPD consisted of a 1334-bp open reading frame (ORF) with a 158-bp intron and the 935-bp upstream and 780-bp downstream regions. The ORF-encoded 391-aa protein (42kDa) showed less than 75% sequence identity to 17 fungal MPDs documented and shared two conserved domains with the fungal MPD family at the N- and C-terminus, respectively. The new enzyme was expressed well in the Luria-Bertani culture of engineered Escherichia coli BL21 by 16-h induction of 0.5 mM isopropyl 1-thio-beta-d-galactopyranoside at 20 degrees C after 5-h growth at 37 degrees C. The purified BbMPD exhibited a high catalytic efficiency (k(cat)/K(m)) of 1.31 x 10(4) mM(-1)s(-1) in the reduction of the highly specific substrate d-fructose-6-phosphate to d-mannitol-1-phosphate. Its activity was maximal at the reaction regime of 37 degrees C and pH 7.0 and was much more sensitive to Cu(2+) and Zn(2+) than to Li(+) and Mn(2+). The results indicate a crucial role of BbMPD in the mannitol biosynthesis of B. bassiana.


Assuntos
Beauveria/genética , Biocatálise , Desidrogenase do Álcool de Açúcar/genética , Desidrogenase do Álcool de Açúcar/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Escherichia coli/genética , Frutosefosfatos/metabolismo , Indicadores e Reagentes/farmacologia , Manitol Fosfatos/metabolismo , Dados de Sequência Molecular , Engenharia de Proteínas , Desidrogenase do Álcool de Açúcar/química , Desidrogenase do Álcool de Açúcar/isolamento & purificação
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