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1.
Electron. j. biotechnol ; 43: 23-31, Jan. 2020. ilus, graf
Artigo em Inglês | LILACS (Américas) | ID: biblio-1087514

RESUMO

Background: Hong Qu glutinous rice wine (HQGRW) is brewed under non-aseptic fermentation conditions, so it usually has a relatively high total acid content. The aim of this study was to investigate the dynamics of the bacterial communities and total acid during the fermentation of HQGRW and elucidate the correlation between total acid and bacterial communities. Results: The results showed that the period of rapid acid increase during fermentation occurred at the early stage of fermentation. There was a negative response between total acid increase and the rate of increase in alcohol during the early fermentation stage. Bacterial community analysis using high-throughput sequencing technology was found that the dominant bacterial communities changed during the traditional fermentation of HQGRW. Both principal component analysis (PCA) and hierarchical clustering analysis revealed that there was a great difference between the bacterial communities of Hong Qu starter and those identified during the fermentation process. Furthermore, the key bacteria likely to be associated with total acid were identified by Spearman's correlation analysis. Lactobacillus, unclassified Lactobacillaceae, and Pediococcus were found, which can make significant contributions to the total acid development (| r| N 0.6 with FDR adjusted P b 0.05), establishing that these bacteria can associate closely with the total acid of rice wine. Conclusions: This was the first study to investigate the correlation between bacterial communities and total acid during the fermentation of HQGRW. These findings may be helpful in the development of a set of fermentation techniques for controlling total acid.


Assuntos
Bactérias/isolamento & purificação , Vinho/microbiologia , Pediococcus/isolamento & purificação , Pediococcus/genética , Pediococcus/metabolismo , Fatores de Tempo , Acetobacter/isolamento & purificação , Acetobacter/genética , Acetobacter/metabolismo , Análise por Conglomerados , Análise de Sequência , Biologia Computacional , Análise de Componente Principal , Fermentação , Microbiota , Concentração de Íons de Hidrogênio , Lactobacillus/isolamento & purificação , Lactobacillus/genética , Lactobacillus/metabolismo
2.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-762473

RESUMO

BACKGROUND: Although the incidence of tuberculosis (TB) is decreasing, cases of multidrug-resistant (MDR) TB and extensively drug-resistant (XDR) TB continue to increase. As conventional phenotype drug susceptibility testing (pDST) takes six to eight weeks, molecular assays are widely used to determine drug resistance. we developed QuantaMatrix Multiplexed Assay Platform (QMAP) MDR/XDR assay (QuantaMatrix Inc., Seoul, Korea) that can simultaneously detect mutations related to both first- and second-line drug resistance (rifampin, isoniazid, ethambutol, fluoroquinolones, second-line injectable drugs, and streptomycin). METHODS: We used 190 clinical Mycobacterium tuberculosis (MTB) strains isolated from Myanmar, compared QMAP and pDST results, and determined concordance rates. Additionally, we performed sequence analyses for discordant results. RESULTS: QMAP results were 87.9% (167/190) concordant with pDST results. In the 23 isolates with discordant results, the QMAP and DNA sequencing results completely matched. CONCLUSIONS: The QMAP MDR/XDR assay can detect all known DNA mutations associated with drug resistance for both MDR- and XDR-MTB strains. It can be used for molecular diagnosis of MDR- and XDR-TB to rapidly initiate appropriate anti-TB drug therapy.


Assuntos
Diagnóstico , DNA , Resistência a Medicamentos , Tratamento Farmacológico , Etambutol , Tuberculose Extensivamente Resistente a Medicamentos , Fluoroquinolonas , Incidência , Isoniazida , Mianmar , Mycobacterium tuberculosis , Fenótipo , Seul , Análise de Sequência , Análise de Sequência de DNA , Tuberculose , Tuberculose Resistente a Múltiplos Medicamentos
3.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-762458

RESUMO

BACKGROUND: Mutations in the quinolone resistance-determining regions (QRDRs) of Acinetobacter baumannii DNA gyrase (gyrA) and topoisomerase IV (parC) are linked to fluoroquinolone (FQ) resistance. We developed a mismatched PCR-restriction fragment length polymorphism (RFLP) assay to detect mutations in the gyrA and parC QRDRs associated with FQ resistance in A. baumannii. METHODS: Based on the conserved sequences of A. baumannii gyrA and parC, two primer sets were designed for mismatched PCR-RFLP to detect mutations in gyrA (codons 83 and 87) and parC (codons 80 and 84) by introducing an artificial restriction enzyme cleavage site into the PCR products. This assay was evaluated using 58 A. baumannii strains and 37 other Acinetobacter strains that have been identified by RNA polymerase β-subunit gene sequence analysis.


Assuntos
Acinetobacter baumannii , Acinetobacter , Sequência Conservada , DNA Girase , DNA Topoisomerase IV , RNA Polimerases Dirigidas por DNA , Reação em Cadeia da Polimerase , Análise de Sequência
4.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-811069

RESUMO

PURPOSE: Different characteristics of airway microbiome in asthmatics may lead to differential immune responses, which in turn cause eosinophilic or neutrophilic airway inflammation. However, the relationships among these factors have yet to be fully elucidated.METHODS: Microbes in induced sputum samples were subjected to sequence analysis of 16S rRNA. Airway inflammatory phenotypes were defined as neutrophils (>60%) and eosinophils (>3%), and inflammation endotypes were defined by levels of T helper (Th) 1 (interferon-γ), Th2 (interleukin [IL]-5 and IL-13), Th-17 (IL-17), and innate Th2 (IL-25, IL-33, and thymic stromal lymphopoietin) cytokines, inflammasomes (IL-1β), epithelial activation markers (granulocyte-macrophage colony-stimulating factor and IL-8), and Inflammation (IL-6 and tumor necrosis factor-α) cytokines in sputum supernatants was assessed by enzyme-linked immunosorbent assay.RESULTS: The numbers of operational taxonomic units were significantly higher in the mixed (n = 21) and neutrophilic (n = 23) inflammation groups than in the paucigranulocytic inflammation group (n = 19; p < 0.05). At the species level, Granulicatella adiacens, Streptococcus parasanguinis, Streptococcus pneumoniae, Veillonella rogosae, Haemophilus parainfluenzae, and Neisseria perflava levels were significantly higher in the eosinophilic inflammation group (n = 20), whereas JYGU_s levels were significantly higher in the neutrophilic inflammation group compared to the other subtypes (P < 0.05). Additionally, IL-5 and IL-13 concentrations were correlated with the percentage of eosinophils (P < 0.05) and IL-13 levels were positively correlated with the read counts of Porphyromonas pasteri and V. rogosae (P < 0.05). IL-1β concentrations were correlated with the percentage of neutrophils (P < 0.05). had a tendency to be positively correlated with the read count of JYGU_s (P = 0.095), and was negatively correlated with that of S. pneumoniae (P < 0.05).CONCLUSIONS: Difference of microbial patterns in airways may induce distinctive endotypes of asthma, which is responsible for the neutrophilic or eosinophilic inflammation in asthma.


Assuntos
Asma , Fatores Estimuladores de Colônias , Citocinas , Ensaio de Imunoadsorção Enzimática , Eosinófilos , Haemophilus parainfluenzae , Inflamassomos , Inflamação , Interleucina-13 , Interleucina-33 , Interleucina-5 , Microbiota , Necrose , Neisseria , Neutrófilos , Fenótipo , Pneumonia , Porphyromonas , Análise de Sequência , Escarro , Streptococcus , Streptococcus pneumoniae , Veillonella
5.
Artigo em Coreano | WPRIM (Pacífico Ocidental) | ID: wprim-816605

RESUMO

BACKGROUND: Rapid and accurate detection of Mycobacterium tuberculosis (MTB) is of primary importance for infection control and selection of anti-tuberculosis drugs. However, most clinical laboratories report MTB complex (MTC) without reporting MTB because MTC comprising MTB, Mycobacterium bovis, Mycobacterium africanum, Mycobacterium microti, Mycobacterium caprae and Mycobacterium pinnipedii have 99.9% similarity at the nucleotide level and identical 16S rRNA sequences. This study was conducted to analyze the species frequency of MTC isolates obtained from clinical specimen.METHODS: Of 310 MTC isolates obtained from clinical samples in a tertiary care hospital from February 2017 to August 2018, MolecuTech Real TB-Taq (YD Diagnostics, Korea) real-time PCR was performed, specifically to detect MTB. For DNA showing MTB negative results by MTB-specific real-time PCR or pyrazinamide-resistant strains, PCR-based MTC typing, spoligotyping, and exact tandem repeat D gene sequencing were performed.RESULTS: All the 310 MTC isolates were identified to be MTB. Two MTB strains of East-African-Indian 4-Vietnam genotype, which have not been reported in Korea, were also found.CONCLUSION: There was no zoonotic tuberculosis in this study. Since we investigated only 310 MTC isolates detected in only one medical institution, multi-center study is needed to accurately know the prevalence of zoonotic tuberculosis in Korea.


Assuntos
DNA , Genótipo , Cabras , Controle de Infecções , Coreia (Geográfico) , Mycobacterium bovis , Mycobacterium tuberculosis , Mycobacterium , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência , Sequências de Repetição em Tandem , Atenção Terciária à Saúde , Tuberculose
6.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-782291

RESUMO

PURPOSE: When influenza viruses are cultured in eggs, amino acid mutations of the hemagglutinin may occur through egg adaptation. On the other hand, when influenza viruses are cultured in animal cells, no antigenic mutation occurs unlike in eggs. Therefore, we examined whether the antigenic mutations actually occurred after passage of H3N2 (A/Texas/50/2012) virus up to 15 times in eggs and MDCK-Sky3851 cells.MATERIALS AND METHODS: Prototype A/Texas/50/2012 (H3N2) influenza virus which was isolated from clinical patient, not passaged in egg, was obtained and propagated using the specific pathogen free egg and the MDCK-Sky3851 cell line up to 15 passage, and the changes in the antigen sequence of the influenza viruses were confirmed by gene sequencing and protein structure analysis.RESULTS: In term of the hemagglutination titer of influenza virus, the reactivity to chicken and guinea pig red blood cell showed different results between egg propagated and cell propagated viruses. In the sequence analysis results for hemagglutinin and neuraminidase, no antigenic mutation was observed throughout all passages when cultured in MDCK-Sky3851 cells. On the other hand, mutations occurred in three amino acid sequences (H156R, G186S, S219F) in hemagglutinin up to 15 passages when cultured in eggs.CONCLUSION: H3N2 influenza virus cultured in eggs could lead mutations in amino acid sequence of hemagglutinin, distinct from the corresponding virus cultured in cells for which no antigenic mutation was observed. These findings suggest that cell culture is a more stable and effective way of production with lower risk of antigenic mutations for the manufacture of influenza vaccines.


Assuntos
Sequência de Aminoácidos , Animais , Técnicas de Cultura de Células , Linhagem Celular , Galinhas , Ovos , Eritrócitos , Cobaias , Mãos , Hemaglutinação , Hemaglutininas , Humanos , Vacinas contra Influenza , Influenza Humana , Neuraminidase , Orthomyxoviridae , Óvulo , Análise de Sequência , Organismos Livres de Patógenos Específicos
7.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-759593

RESUMO

BACKGROUND: The recent expansion of knowledge about various ABO alleles has led to the need for a comprehensive measure to cover the numerous polymorphisms dispersed in the ABO gene. A few studies have examined the diversity of the O allele compared to A or B subgroup alleles, resulting in antigenic changes. This study investigated the relationship between the serologic and molecular genetic characteristics of the O alleles in the Korean population. METHODS: One hundred and five samples from healthy blood group O subjects were selected randomly. The isoagglutinin titer was measured using a tube agglutination and gel microcolumn assay. The ABO alleles were analyzed by sequencing exons 6 and 7 of the ABO gene. When the origin of a heterozygous nucleotide sequence was ambiguous, it was separated into a single allele using mono-allele amplification or cloning. RESULTS: The median IgM isoagglutinin titer was eight. In contrast, the median IgG anti-A and anti-B isoagglutinin titers were 64 and 32, respectively. The IgG isoagglutinin titer showed a significant increase with age (P<0.0001). Six O alleles were observed in 105 blood group O populations by sequencing. The O01 and O02 alleles were common (0.57, 0.36). Three rare O alleles (O04, O05, and O06) and one novel non-deletional O allele were found. CONCLUSION: The distribution of isoagglutinin titers of blood group O and the genetic frequency of O alleles in this study would form the basis of the development and interpretation of ABO genotyping and serologic workup in the Korean population.


Assuntos
Aglutinação , Alelos , Sequência de Bases , Células Clonais , Clonagem de Organismos , Éxons , Imunoglobulina G , Imunoglobulina M , Biologia Molecular , Análise de Sequência
8.
Artigo em Coreano | WPRIM (Pacífico Ocidental) | ID: wprim-766742

RESUMO

Autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS), which is caused by mutations in SACS gene, is a very rare neurodegenerative disorder characterized by the clinical triad of early onset cerebellar ataxia, pyramidal tract features, and sensorimotor polyneuropathy. Herein, we report a 35-year-old Korean male who presented with gait disturbance and lower extremity weakness. Neuroimaging and ophthalmologic evaluation revealed features consistent with ARSACS. Mutation in SACS gene was demonstrated in clinical exome sequence analysis and the patient was finally diagnosed as ARSACS.


Assuntos
Adulto , Ataxia , Ataxia Cerebelar , Exoma , Marcha , Humanos , Extremidade Inferior , Masculino , Espasticidade Muscular , Doenças Neurodegenerativas , Neuroimagem , Polineuropatias , Tratos Piramidais , Análise de Sequência , Degenerações Espinocerebelares
9.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-760648

RESUMO

OBJECTIVE: Human papillomaviruses (HPVs) are among the agents responsible for infection and cancer of the skin and mucous membranes in the human body. The aim of this study was to investigate the frequency and type distribution of HPVs in married female patients with gynecological complaints, who had visited the Maternity Hospital in Erzurum, Turkey. METHODS: In this study, 263 cervical swab samples were taken from married women using the Pap smear method and were investigated for positive reactivity against HPV. The L1 gene region of HPV was investigated using molecular methods. For this purpose, polymerase chain reaction (PCR) assays and sequence analysis of positive samples were performed. Phylogenetic analyses were performed using a bioinformatics approach after sequencing. RESULTS: HPV-DNA was detected in 17 (6.5%) samples. Highest positive reactivity to HPV-DNA was found in the 35–44 age group at 9.2%. Fourteen out of seventeen positive samples were included in the phylogenetic analysis. All isolates clustered in the Alphapapillomavirus genus. Six samples were found to be HPV 70 positive, four were HPV 16 positive, and the rest were HPV 54, 72, 81, and 114 positive. When genotyping data were evaluated according to the risk group, we found that 28.6% of the 14 samples were found to be high risk-HPV, and 71.4% were low risk-HPV. CONCLUSIONS: As per our knowledge, this is the first report on the phylogenetic analysis of HPV genotypes isolated from women in Turkey. The prevalence of low- and-high risk HPV was determined in married women in Erzurum, and these results contribute to the epidemiological data on the distribution of HPV types for this region.


Assuntos
Alphapapillomavirus , Biologia Computacional , Feminino , Genótipo , Maternidades , Corpo Humano , Papillomavirus Humano 16 , Humanos , Métodos , Membrana Mucosa , Teste de Papanicolaou , Infecções por Papillomavirus , Reação em Cadeia da Polimerase , Prevalência , Análise de Sequência , Neoplasias Cutâneas , Turquia , Esfregaço Vaginal
10.
Mycobiology ; : 154-164, 2019.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-760544

RESUMO

Four strains of Penicillium and Talaromyces species are described and illustrated in an inventory of fungal species belonging to Eurotiales. The strains, CNUFC-DDS17-1, CNUFC-DDS27-1, CNUFC-PTM72-1, and CNUFC-YJW3-31, were isolated from soil and freshwater samples from South Korea. Based on their morphological characteristics and sequence analyses by the combined β-tubulin and calmodulin gene, the CNUFC-DDS17-1, CNUFC-DDS27-1, CNUFC-PTM72-1, and CNUFC-YJW3-31 isolates were identified as Penicillium pasqualense, Penicillium sanguifluum, Talaromyces apiculatus, and Talaromyces liani, respectively. The designated strains were found to represent a previously undescribed species of Korean fungal biota. In this study, detailed morphological descriptions and phylogenetic relationships of these species are provided.


Assuntos
Biota , Calmodulina , Eurotiales , Água Doce , Coreia (Geográfico) , Penicillium , Análise de Sequência , Solo , Talaromyces
11.
Mycobiology ; : 165-172, 2019.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-760543

RESUMO

Species that belong to Penicillium section Sclerotiora are commonly found in various terrestrial environments, but only a few have been reported in marine environments. Because the number of Penicillium species reported in marine environments is increasing, we investigated the diversity of Penicillium section Sclerotiora in marine environments in Korea. Based on sequence analyses of β-tubulin and calmodulin loci, 21 strains of section Sclerotiora were identified as P. bilaiae, P. daejeonium, P. exsudans, P. herquei, P. cf. guanacastense, P. mallochii, P. maximae, and P. viticola. Three of them were confirmed as new to Korea: P. exsudans, P. mallochii, and P. maximae. Here, we have provided detailed morphological descriptions of these unrecorded species.


Assuntos
Calmodulina , Coreia (Geográfico) , Penicillium , Filogenia , Análise de Sequência
12.
Laboratory Medicine Online ; : 177-180, 2019.
Artigo em Coreano | WPRIM (Pacífico Ocidental) | ID: wprim-760497

RESUMO

Catabacter hongkongensis is an anaerobic gram-positive coccobacillus that was first isolated in Hong Kong. It is infectious and causes high mortality in patients with rare but underlying diseases. Alistipes indistinctus is an anaerobic gram-negative coccobacillus. This bacterium is a common member of the human intestinal microbiota. We report a case of C. hongkongensis and A. indistinctus isolated from blood cultures of a patient with acute appendicitis. A 35-year-old female patient with no specific medical history was admitted to the hospital due to abdominal pain, vomiting, nausea, and diarrhea experienced on the day before admission. On admission, laboratory tests revealed leukocytosis, neutropenia, and elevated C–reactive protein and procalcitonin levels. Following an abdominal computed tomography showing acute appendicitis with suspected perforation, emergency surgery was performed. Growth was observed in two anaerobic blood culture bottles after four days. After further culturing of the bacteria on Brucella Blood Agar, two types of bacteria were obtained. The two bacterial isolates, one gram-positive and one gram-negative, were unable to be identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Thus, 16S rRNA gene sequence analysis was performed, resulting in identification of the bacteria as C. hongkongensis and A. indistinctus. The patient was administered antibiotics and discharged two days after surgery. Although MALDI-TOF MS enables fast and accurate identification of bacteria, C. hongkongensis and A. indistinctus were not listed in the spectral library, and 16S rRNA gene sequence analysis was useful for identifying the two bacteria.


Assuntos
Dor Abdominal , Adulto , Ágar , Antibacterianos , Apendicite , Bactérias , Brucella , Diarreia , Emergências , Feminino , Microbioma Gastrointestinal , Genes de RNAr , Hong Kong , Humanos , Leucocitose , Espectrometria de Massas , Mortalidade , Náusea , Neutropenia , Análise de Sequência , Vômito
13.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-760353

RESUMO

Avian malaria is one of the most important general blood parasites of poultry in Southeast Asia. Plasmodium (P.) juxtanucleare causes avian malaria in wild and domestic fowl. This study aimed to identify and characterize the Plasmodium species infecting in Thai native fowl. Blood samples were collected for microscopic examination, followed by detection of the Plasmodium cox I gene by using PCR. Five of the 10 sampled fowl had the desired 588 base pair amplicons. Sequence analysis of the five amplicons indicated that the nucleotide and amino acid sequences were homologous to each other and were closely related (100% identity) to a P. juxtanucleare strain isolated in Japan (AB250415). Furthermore, the phylogenetic tree of the cox I gene showed that the P. juxtanucleare in this study were grouped together and clustered with the Japan strain. The presence of P. juxtanucleare described in this study is the first report of P. juxtanucleare in the Thai native fowl of Thailand.


Assuntos
Sequência de Aminoácidos , Animais , Ásia Sudeste , Grupo com Ancestrais do Continente Asiático , Pareamento de Bases , Citocromos c , Citocromos , Complexo IV da Cadeia de Transporte de Elétrons , Humanos , Japão , Malária Aviária , Parasitos , Plasmodium , Reação em Cadeia da Polimerase , Aves Domésticas , Análise de Sequência , Tailândia , Árvores
14.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-764311

RESUMO

BACKGROUND: Gut microbiota is closely associated with development and exacerbation of inflammatory bowel diseases (IBD). The aim of this study was to investigate differences in gut microbiota depending on sex and changes of gut microbiota during IBD developments. METHODS: 16s rRNA metagenomic sequencing was performed for fecal materials from 8-week-old wild type (WT) and interleukin 10 (IL-10) knockout (KO) C57BL/6 mice of both sexes. Diversity indices, relative abundance of microbiota, and linear discriminant analysis effect size were examined to compare microbial communities between groups. Clustering of groups was performed by principal coordinates analysis (PCoA) and unweighted pair group method with arithmetic mean (UPGMA). Functional capabilities of microbiota were estimated using phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) based on Kyoto Encyclopedia of Genes and Genomes database. RESULTS: PCoA and UPGMA tree analysis of beta-diversity demonstrated significant differences in gut microbiota between male and female groups of WT mice, but not of IL-10 KO mice. Firmicutes to Bacteroides ratio was higher in male group than that in female group in both WT mice and IL-10 KO mice. Phylum Proteobacteria significantly increased in female IL-10 KO mice than that in female WT mice. At species level, Lactobacillus murinus, Bacteroides acidifaciens, and Helicobacter hepaticus significantly increased in IL-10 KO mice than in WT mice. The relative abundance of beta-glucuronidase (K01195) was higher in female IL-10 KO mice than that in female WT mice by PICRUSt. CONCLUSIONS: Our results suggest that microbiota-host interactions might differ between sexes during development of IBD.


Assuntos
Animais , Bacteroides , Feminino , Firmicutes , Microbioma Gastrointestinal , Genoma , Glucuronidase , Helicobacter hepaticus , Humanos , Doenças Inflamatórias Intestinais , Interleucina-10 , Lactobacillus , Masculino , Metagenômica , Métodos , Camundongos , Microbiota , Proteobactérias , Análise de Sequência , Caracteres Sexuais , Árvores
15.
Artigo em Coreano | WPRIM (Pacífico Ocidental) | ID: wprim-816602

RESUMO

BACKGROUND: Candida auris was first isolated from the ears of Japanese and Korean patients. However, the prevalence of yeast isolates from ear cultures and their antifungal susceptibility profiles in these nations remain unclear.METHODS: We assessed yeast isolates recovered from ear cultures from a university hospital in Korea over a 4-year period from January 2014 to December 2017. Species identification was performed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and/or sequence analysis. Antifungal minimal inhibitory concentrations (MICs) were determined using the broth microdilution method of the Clinical and Laboratory Standards Institute.RESULTS: Among 81 non-duplicate isolates from ear cultures, Cadida parapsilosis was the most frequently detected yeast species (34.6%), followed by C. auris (28.4%), Candida metapsilosis (9.9%), Candida orthopsilosis (8.6%), Candida albicans (7.4%), and others (11.1%). The MICs of the isolates were 0.125 to > 64 µg/mL, ≤0.03 to 4 µg/mL, 0.25 to 1 µg/mL, 0.125 to 1 µg/mL, and ≤0.03 to 2 µg/mL for fluconazole, voriconazole, amphotericin B, caspofungin, and micafungin, respectively. Of the 81 isolates, 44.4% (36/81) showed decreased susceptibility to fluconazole (MIC ≥4 µg/mL). Of the 23 C. auris isolates, 19 (82.6%) had a fluconazole MIC of ≥32 µg/mL. None of the isolates showed resistance to amphotericin B or echinocandins. Most of these patients suffered from chronic otitis media (84%).CONCLUSION: Candida parapsilosis complex and C. auris were the yeast species identified most frequently from ear cultures and they exhibited a high rate of fluconazole non-susceptibility, particularly C. auris.


Assuntos
Anfotericina B , Grupo com Ancestrais do Continente Asiático , Candida , Candida albicans , Orelha , Equinocandinas , Fluconazol , Humanos , Coreia (Geográfico) , Espectrometria de Massas , Métodos , Otite Média , Prevalência , Análise de Sequência , Voriconazol , Leveduras
16.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-816599

RESUMO

We report a case of cellulitis caused by a novel Cupriavidus species identified using whole-genome sequence analysis. Subcutaneous tissue biopsies from the left lower leg of a 67-year-old man who suffered from cellulitis were cultured. Round, convex, gray and non-hemolytic colonies were recovered after 72-h incubation. 16S rRNA sequence analysis showed 98.6% similarity with Cupriavidus basilensis DSM 11853(T) in the NCBI database and 99.9% similarity with C. basilensis KF708 in the EzBioCloud database. Genomic analysis using the MiSeq platform (Illumina, USA) and the TrueBac ID database (ChunLab, Korea) revealed that the average nucleotide identity (ANI) of this strain with C. basilensis DSM 11853(T) was 87.6%. The patient was treated with oral cefditoren pivoxil for 9 weeks. This study is the first to report cellulitis caused by Cupriavidus species strain J1218.


Assuntos
Idoso , Biópsia , Celulite (Flegmão) , Cupriavidus , Genoma , Humanos , Perna (Membro) , Análise de Sequência , Tela Subcutânea
17.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wprim-773406

RESUMO

OBJECTIVE@#People in Western Africa suffer greatly from febrile jaundice, which is caused by a variety of pathogens. However, yellow fever virus (YFV) is the only pathogen under surveillance in Sierra Leone owing to the undeveloped medical and public health system there. Most of the results of YFV identification are negative. Elucidation of the pathogen spectrum is required to reduce the prevalence of febrile jaundice.@*METHODS@#In the present study, we used Ion Torrent semiconductor sequencing to profile the pathogen spectrum in archived YFV-negative sera from 96 patients in Sierra Leone who presented with unexplained febrile jaundice.@*RESULTS@#The most frequently identified sequencing reads belonged to the following pathogens: cytomegalovirus (89.58%), Epstein-Barr virus (55.21%), hepatitis C virus (34.38%), rhinovirus (28.13%), hepatitis A virus (20.83%), coxsackievirus (10.42%), Ebola virus (8.33%), hepatitis E virus (8.33%), lyssavirus (4.17%), leptospirosis (4.17%), chikungunya virus (2.08%), Crimean-Congo hemorrhagic fever virus (1.04%), and hepatitis B virus (1.04%).@*CONCLUSION@#The distribution of sequencing reads suggests a broader spectrum of pathogens for consideration in clinical diagnostics and epidemiological surveillance in Sierra Leone.


Assuntos
Adolescente , Adulto , Estudos de Casos e Controles , Feminino , Febre , Epidemiologia , Virologia , Humanos , Icterícia , Epidemiologia , Virologia , Masculino , Análise de Sequência , Serra Leoa , Epidemiologia , Adulto Jovem
18.
Electron. j. biotechnol ; 32: 6-12, Mar. 2018. tab, graf, ilus
Artigo em Inglês | LILACS (Américas) | ID: biblio-1022493

RESUMO

Background: Hydrophobins are small proteins secreted by filamentous fungi, which show a highly surface activity. Because of the signally self-assembling abilities and surface activities, hydrophobins were considered as candidates in many aspects, for example, stabilizing foams and emulsions in food products. Lentinus tuber-regium, known as tiger milk mushroom, is both an edible and medicinal sclerotium-producing mushroom. Up to now, the hydrophobins of L. tuber-regium have not been identified. Results: In this paper, a Class I hydrophobin gene, Ltr.hyd, was cloned from L. tuber-regium and expressed in the yeast-like cells of Tremella fuciformis mediated by Agrobacterium tumefaciens. The expression vector pGEH-GH was under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter. The integration of Ltr.hyd into the genome of T. fuciformis was confirmed by PCR, Southern blot, fluorescence observation and quantitative real-time PCR (qRT-PCR). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that recombinant hydrophobin rLtr.HYD with an expected molecular mass of 13 kDa was extracted. The yield of rLtr.HYD was 0.66 mg/g dry weight. The emulsifying activity of rLtr.HYD was better than the typical food emulsifiers sodium caseinate and Tween 20. Conclusions: We evaluated the emulsifying property of hydrophobin Ltr.HYD, which can be potentially used as a food emulsifier.


Assuntos
Basidiomycota/metabolismo , Proteínas Fúngicas/genética , Lentinula/genética , Lentinula/metabolismo , Transformação Genética , Basidiomycota/enzimologia , Leveduras , Proteínas Fúngicas/metabolismo , Southern Blotting , Clonagem Molecular , Agrobacterium tumefaciens/metabolismo , Análise de Sequência , Emulsificantes , Eletroforese em Gel de Poliacrilamida , Reação em Cadeia da Polimerase em Tempo Real , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Microscopia de Fluorescência
19.
Electron. j. biotechnol ; 32: 47-54, Mar. 2018. tab, ilus, graf
Artigo em Inglês | LILACS (Américas) | ID: biblio-1022746

RESUMO

Background: Cathepsin C (CTSC) (dipeptidyl peptidase I, DPPI), is a member of the papain superfamily of cysteine proteases and involves in a variety of host reactions. However, the information of CTST in Chinese giant salamander (Andrias davidianus), an amphibian species with important evolutionary position and economic values, remained unclear. Results: The full-length salamander CTSC cDNA contained a 96 bp of 5'-UTR, a 1392 bp of ORF encoding 463 amino acids, and a 95 bp of 3'-UTR. The salamander CTSC possessed several sequence features similar to other reported CTSCs such as a signal peptide, a propeptide and a mature peptide. The active site triad of Cys, His and Asn were also found existing in salamander CTSC. Salamander CTSC mRNA was constitutively expressed in all the examined tissues with significantly variant expression level. The highest expression of CTSC was in intestine, followed with stomach, spleen, lung and brain. Following Aeromonas hydrophila infection for 12 h, salamander CTSC was significantly up-regulated in several tissues including lung, spleen, brain, kidney, heart, stomach and skin. Conclusion: CTSC plays roles in the immune response to bacterial infection, which provided valuable information for further studying the functions of CTSC in salamander.


Assuntos
Animais , Urodelos/genética , Urodelos/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Catepsina C/imunologia , Urodelos/microbiologia , Infecções por Bactérias Gram-Negativas/imunologia , Clonagem Molecular , Aeromonas hydrophila/fisiologia , Análise de Sequência , DNA Complementar , Catepsina C/genética , Catepsina C/metabolismo , Transcrição Reversa , Imunidade Inata/genética
20.
Electron. j. biotechnol ; 31: 10-16, Jan. 2018. graf, tab, ilust
Artigo em Inglês | LILACS (Américas) | ID: biblio-1022030

RESUMO

Background: Biodegradation is a reliable approach for efficiently eliminating persistent pollutants such as chlorpyrifos. Despite many bacteria or fungi isolated from contaminated environment and capable of degrading chlorpyrifos, limited enzymes responsible for its degradation have been identified, let alone the catalytic mechanism of the enzymes. Results: In present study, the gene cpd encoding a chlorpyrifos hydrolase was cloned by analysis of genomic sequence of Paracoccus sp. TRP. Phylogenetic analysis and BLAST indicated that CPD was a novel member of organophosphate hydrolases. The purified CPD enzyme, with conserved catalytic triad (Ser155-Asp251-His281) and motif Gly-Asp-Ser-Ala-Gly, was significantly inhibited by PMSF, a serine modifier. Molecular docking between CPD and chlorpyrifos showed that Ser155 was adjacent to chlorpyrifos, which indicated that Ser155 may be the active amino acid involved in chlorpyrifos degradation. This speculation was confirmed by site-directed mutagenesis of Ser155Ala accounting for the decreased activity of CPD towards chlorpyrifos. According to the key role of Ser155 in chlorpyrifos degradation and molecular docking conformation, the nucleophilic catalytic mechanism for chlorpyrifos degradation by CPD was proposed. Conclusion: The novel enzyme CPD was capable of hydrolyze chlorpyrifos and Ser155 played key role during degradation of chlorpyrifos.


Assuntos
Paracoccus/enzimologia , Clorpirifos/metabolismo , Esterases/metabolismo , Organofosfatos/metabolismo , Biodegradação Ambiental , Catálise , Mutagênese , Clonagem Molecular , Análise de Sequência , Esterases/isolamento & purificação , Esterases/genética , Hidrólise , Metais/metabolismo
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