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1.
Phytother Res ; 37(10): 4722-4739, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37443453

RESUMO

Epithelial ovarian cancer (EOC) is the most common and fatal subtype of ovarian malignancies, with no effective therapeutics available. Our previous studies have demonstrated extraordinary suppressive efficacy of enterolactone (ENL) on EOC. A chemotherapeutic agent, trabectedin (Trabe), is shown to be effective on ovarian cancer, especially when combined with other therapeutics, such as pegylated liposomal doxorubicin or oxaliplatin. Thrombospondin 1 (THBS1), a kind of matrix glycoprotein, plays important roles against cancer development through inhibiting angiogenesis but whether it is involved in the suppression of EOC by ENL or Trabe remains unknown. To test combined suppressive effects of ENL and Trabe on EOC and possible involvement of THBS1 in the anticancer activities of ENL and Trabe. The EOC cell line ES-2 was transfected with overexpressed THBS1 by lentivirus vector. We employed tube formation assay to evaluate the anti-angiogenesis activity of ENL and of its combined use with Trabe after THBS1 overexpression and established drug intervention and xenograft nude mouse cancer models to assess the in vivo effects of the hypothesized synergistic suppression between the agents and the involvement of THBS1. Mouse fecal samples were collected for 16S rDNA sequencing and microbiota analysis. We detected strong inhibitory activities of ENL and Trabe against the proliferation and migration of cancer cells and observed synergistic effects between ENL and Trabe in suppressing EOC. ENL and Trabe, given either separately or in combination, could suppress the tube formation capability of human microvascular endothelial cells, and this inhibitory effect became even stronger with THBS1 overexpression. In the ENL plus Trabe combination group, the expression of tissue inhibitor of metalloproteinases 3 and cluster of differentiation 36 was both upregulated, whereas matrix metalloproteinase 9, vascular endothelial growth factor, and cluster of differentiation 47 were all decreased. With the overexpression of THBS1, the results became even more pronounced. In animal experiments, combined use of ENL and Trabe showed superior inhibitory effects to either single agent and significantly suppressed tumor growth, and the overexpression of THBS1 further enhanced the anti-cancer activities of the drug combination group. ENL and Trabe synergistically suppress EOC and THBS1 could remarkably facilitate the synergistic anticancer effects of ENL and Trabe.


Assuntos
Neoplasias Ovarianas , Trombospondina 1 , Animais , Camundongos , Humanos , Feminino , Carcinoma Epitelial do Ovário , Trabectedina/uso terapêutico , Trombospondina 1/uso terapêutico , Fator A de Crescimento do Endotélio Vascular , Células Endoteliais/metabolismo , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética
2.
Invest New Drugs ; 38(5): 1373-1382, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32734371

RESUMO

Cancer is one of the leading causes of death worldwide, but effective therapies remain the topic of many research activities. Many recent studies have thus focused on particular gut microbiota due to their important roles in treating cancers, but very few microbes of therapeutic value have been reported. In this study, we isolated four bacterial strains, BY38, BY40, BY43 and BY45, from the fecal specimens of healthy individuals and cancer patients. The treatment of cancer cells with the products of these cultured bacteria induced significant inhibitory effects on the proliferation of ovarian cancer cells and colorectal cancer cells in a dose-dependent manner. A phylogenetic analysis showed that the four anticancer strains belong to the genus Bacillus, and flow cytometry assays indicated that the inhibitory effects might be achieved through the induction of cell apoptosis. These results suggest that these bacteria could be novel and promising anticancer agents against cancers.


Assuntos
Antineoplásicos/farmacologia , Bacillus/metabolismo , Produtos Biológicos/farmacologia , Microbioma Gastrointestinal , Neoplasias/tratamento farmacológico , Adulto , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Fezes/microbiologia , Genoma Bacteriano , Humanos , Pessoa de Meia-Idade , Filogenia , Sequenciamento Completo do Genoma
3.
BMC Genomics ; 20(1): 930, 2019 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-31801462

RESUMO

BACKGROUND: Salmonella bongori infect mainly cold-blooded hosts, but infections by S. bongori in warm-blooded hosts have been reported. We hypothesized that S. bongori might have diverged into distinct phylogenetic lineages, with some being able to infect warm-blooded hosts. RESULTS: To inspect the divergence status of S. bongori, we first completely sequenced the parakeet isolate RKS3044 and compared it with other sequenced S. bongori strains. We found that RKS3044 contained a novel T6SS encoded in a pathogenicity island-like structure, in addition to a T6SS encoded in SPI-22, which is common to all S. bongori strains so far reported. This novel T6SS resembled the SPI-19 T6SS of the warm-blooded host infecting Salmonella Subgroup I lineages. Genomic sequence comparisons revealed different genomic sequence amelioration events among the S. bongori strains, including a unique CTAG tetranucleotide degeneration pattern in RKS3044, suggesting non-overlapping gene pools between RKS3044 and other S. bongori lineages/strains leading to their independent accumulation of genomic variations. We further proved the existence of a clear-cut genetic boundary between RKS3044 and the other S. bongori lineages/strains analyzed in this study. CONCLUSIONS: The warm-blooded host-infecting S. bongori strain RKS3044 has diverged with distinct genomic features from other S. bongori strains, including a novel T6SS encoded in a previously not reported pathogenicity island-like structure and a unique genomic sequence degeneration pattern. These findings alert cautions about the emergence of new pathogens originating from non-pathogenic ancestors by acquiring specific pathogenic traits.


Assuntos
Ilhas Genômicas , Periquitos/microbiologia , Salmonella/classificação , Sequenciamento Completo do Genoma/métodos , Animais , Evolução Molecular , Especiação Genética , Tamanho do Genoma , Genoma Bacteriano , Humanos , Filogenia , Salmonella/genética , Salmonella/patogenicidade , Fatores de Virulência/genética
4.
Mol Genet Genomics ; 294(3): 597-605, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30710177

RESUMO

Antimicrobial resistance makes pathogenic bacteria hard to control, but little is known about the general processes of resistance gain or loss. Here, we compared distinct S. typhimurium DT104 strains resistant to zero, two, five, or more of the tested antimicrobials. We found that common resistance phenotypes could be encoded by distinct genes, on SGI-1 or plasmid. We also demonstrated close clonality among all the tested non-resistant and differently resistant DT104 strains, demonstrating dynamic acquisition or loss (by total deletion or gradual decaying of multi-drug resistance gene clusters) of the genetic traits. These findings reflect convergent processes to make the bacteria resistant to multiple antimicrobials by acquiring the needed traits from stochastically available origins. When the antimicrobial stress is absent, the resistance genes may be dropped off quickly, so the bacteria can save the cost for maintaining unneeded genes. Therefore, this work reiterates the importance of strictly controlled use of antimicrobials.


Assuntos
Adaptação Fisiológica/genética , Farmacorresistência Bacteriana Múltipla/genética , Evolução Molecular , Salmonella typhimurium/genética , Estresse Fisiológico , Adaptação Fisiológica/efeitos dos fármacos , Antibacterianos/farmacologia , Sequência de Bases , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Genes Bacterianos/genética , Genoma Bacteriano/genética , Filogenia , Plasmídeos/classificação , Plasmídeos/genética , Salmonella typhimurium/classificação , Salmonella typhimurium/efeitos dos fármacos , Homologia de Sequência do Ácido Nucleico
5.
BMC Microbiol ; 14: 157, 2014 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-24928416

RESUMO

BACKGROUND: Acquisition of exogenous genetic material is a key event in bacterial speciation. It seems reasonable to assume that recombination of the incoming DNA into genome would be more efficient with higher levels of relatedness between the DNA donor and recipient. If so, bacterial speciation would be a smooth process, leading to a continuous spectrum of genomic divergence of bacteria, which, however, is not the case as shown by recent findings. The goal of this study was todetermine if DNA transfer efficiency is correlated with the levels of sequence identity. RESULTS: To compare the relative efficiency of exogenous DNA acquisition among closely related bacteria, we carried out phage-mediated transduction and plasmid-mediated transformation in representative Salmonella strains with different levels of relatedness. We found that the efficiency was remarkably variable even among genetically almost identical bacteria. Although there was a general tendency that more closely related DNA donor-recipient pairs had higher transduction efficiency, transformation efficiency exhibited over a thousand times difference among the closely related Salmonella strains. CONCLUSION: DNA acquisition efficiency is greatly variable among bacteria that have as high as over 99% identical genetic background, suggesting that bacterial speciation involves highly complex processes affected not only by whether beneficial exogenous DNA may exist in the environment but also the "readiness" of the bacteria to accept it.


Assuntos
DNA/genética , Recombinação Genética , Salmonella/genética , Transdução Genética , DNA/metabolismo , DNA Bacteriano/química , DNA Bacteriano/genética , Especiação Genética , Dados de Sequência Molecular , Salmonella/metabolismo , Análise de Sequência de DNA , Transformação Bacteriana
6.
Phys Chem Chem Phys ; 16(37): 19972-83, 2014 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-25159551

RESUMO

We report the hydrostatic compression studies of the ß-polymorph of a cyclotetramethylene tetranitramine (HMX) energetic molecular crystal using DFT-D2, a first-principles calculation based on density functional theory (DFT) with van der Waals (vdW) corrections. The molecular structure, mechanical properties, electronic properties, and equations of state of ß-HMX are investigated. For the first time, we predict the elastic constants of ß-HMX using DFT-D2 studies. The equations of state under hydrostatic compression are studied for pressures up to 100 GPa. We found that the N-N bonds along the minor axis are responsible for the sensitivity of ß-HMX. The analysis of the charge distribution shows that the electronic charge is transferred from hydrogen atoms to nitro groups with the amount of 0.131 and 0.064e for the nitro groups along the minor axis and major axis, respectively, when pressure changes from 0 GPa to 100 GPa. The electronic energy band gap changes from direct at a pressure of 0 GPa to indirect at a pressure of 50 GPa and higher. The band gap decreases with respect to an increase in pressure, implying that the impact sensitivity increases with compression. Our study suggests that the van der Waals interactions are critically important in modeling the mechanical properties of this molecular crystal.

7.
Genomics ; 102(4): 331-7, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23933189

RESUMO

The deadly human typhoid agent was initially classified as a species called Salmonella typhi but later reclassified as a serovar of Salmonella enterica together with other pathogenically diverse serovars. The dynamic changes of Salmonella taxonomy reflect the need to clarify the phylogenetic status of the Salmonella serovars: are they discrete lineages or variants of a genetic lineage? To answer this question, we compared S. typhi and other Salmonella serotypes. We found that the S. typhi and Salmonella typhimurium strains had over 90% and ca. 80%, respectively, of their genes identical; however, between S. typhi and S. typhimurium, this percentage dropped to 6%, suggesting the existence of genetic boundaries between them. We conclude that S. typhi and the other compared Salmonella serovars have developed into distinct lineages circumscribed by the genetic boundary. This concept and methods may be used to delineate other Salmonella serotypes, many of which are polyphyletic, needing differentiation.


Assuntos
Genes Bacterianos , Salmonella enterica/classificação , Salmonella enterica/genética , Febre Tifoide/microbiologia , Evolução Molecular , Especiação Genética , Humanos , Dados de Sequência Molecular , Filogenia , Salmonella typhi/classificação , Salmonella typhi/genética , Salmonella typhimurium/classificação , Salmonella typhimurium/genética , Homologia de Sequência , Sorotipagem , Febre Tifoide/patologia
8.
Int Med Case Rep J ; 17: 167-175, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38504721

RESUMO

Introduction: Pancreatic cancer (PC) is among the deadliest malignancies. Kidney cancer (KC) is a common malignancy globally. Chemo- or radio-therapies are not very effective to control PC or KC, and overdoses often cause severe site reactions to the patients. As a result, novel treatment strategies with high efficacy but without toxic side effects are urgently desired. Secoisolariciresinol diglucoside (SDG) belongs to plant lignans with potential anticancer activities, but clinical evidence is not available in PC or KC treatment. Patient Concerns: We report a rare case of an 83-year-old female patient with pancreatic and kidney occupying lesions that lacked the conditions to receive surgery or chemo- or radiotherapy. Diagnosis: Pancreatic and kidney cancers. Interventions: We gave dietary SDG to the patient as the only therapeutics. Outcomes: SDG effectively halted progression of both PC and KC. All clinical manifestations, including bad insomnia, loss of appetite, stomach symptoms, and skin itching over the whole body, all disappeared. The initial massive macroscopic hematuria became microscopic and infrequent, and other laboratory results also gradually returned to normal. Most of the cancer biomarkers, initially high such as CEA, CA199, CA724, CA125, came down rapidly, among which CA199 changed most radically. This patient has had progression-free survival of one year so far. Conclusion: These results demonstrate the potent inhibitory effects of SDG on PC and KC of this patient and provide promising novel therapeutics for refractory malignant tumors.

9.
BMC Genomics ; 14: 489, 2013 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-23865772

RESUMO

BACKGROUND: Bacteria are currently classified into arbitrary species, but whether they actually exist as discrete natural species was unclear. To reveal genomic features that may unambiguously group bacteria into discrete genetic clusters, we carried out systematic genomic comparisons among representative bacteria. RESULTS: We found that bacteria of Salmonella formed tight phylogenetic clusters separated by various genetic distances: whereas over 90% of the approximately four thousand shared genes had completely identical sequences among strains of the same lineage, the percentages dropped sharply to below 50% across the lineages, demonstrating the existence of clear-cut genetic boundaries by a steep turning point in nucleotide sequence divergence. Recombination assays supported the genetic boundary hypothesis, suggesting that genetic barriers had been formed between bacteria of even very closely related lineages. We found similar situations in bacteria of Yersinia and Staphylococcus. CONCLUSIONS: Bacteria are genetically isolated into discrete clusters equivalent to natural species.


Assuntos
Bactérias/genética , Variação Genética/genética , Genômica/métodos , Nucleotídeos/genética , DNA Bacteriano/genética , Evolução Molecular , Filogenia , Recombinação Genética/genética
10.
Mol Genet Genomics ; 288(11): 549-57, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23933962

RESUMO

DT104 emerged as a new branch of Salmonella typhimurium with resistance to multiple antimicrobials. To reveal some general genomic features of DT104 for clues of evolutionary events possibly associated with the emergence of this relatively new type of this pathogen, we mapped 11 independent DT104 strains and compared them with non-DT104 S. typhimurium strains. We found that all 11 DT104 strains contained three insertions absent in non-DT104 strains, i.e., the previously reported ST104, ST104B and ST64B. However, SGI-1, a genomic island known to be responsible for DT104 multidrug resistance, was not present in all DT104 strains examined in this study: one DT104 strain did not contain SGI-1 but carried a 144 kb plasmid, suggesting possible evolutionary relationships between the two DNA elements in the development of antimicrobial resistance.


Assuntos
Genoma Bacteriano/genética , Genômica , Infecções por Salmonella/microbiologia , Salmonella typhimurium/genética , Evolução Biológica , Mapeamento Cromossômico , DNA Bacteriano/química , DNA Bacteriano/genética , Desoxirribonucleases de Sítio Específico do Tipo II , Farmacorresistência Bacteriana Múltipla/genética , Endodesoxirribonucleases , Rearranjo Gênico , Ilhas Genômicas/fisiologia , Plasmídeos/genética , Especificidade da Espécie
11.
Antonie Van Leeuwenhoek ; 103(4): 797-807, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23224438

RESUMO

Pseudomonas aeruginosa, a wide-spread opportunistic pathogen, often complicates clinical treatments due to its resistance to a large variety of antimicrobials, especially in immune compromised patients, occasionally leading to death. However, the resistance to antimicrobials varies greatly among the P. aeruginosa isolates, which raises a question on whether some sub-lineages of P. aeruginosa might have greater potential to develop antimicrobial resistance than others. To explore this question, we divided 160 P. aeruginosa isolates collected from cities of USA and China into distinct genotypes using I-CeuI, a special endonuclease that had previously been proven to reveal phylogenetic relationships among bacteria reliably due to the highly conserved 26-bp recognition sequence. We resolved 10 genotypes by I-CeuI analysis and further divided them into 82 sub-genotypes by endonuclease cleavage with SpeI. Eight of the 10 genotypes contained both multi-drug resistant (MDR) and less resistant isolates based on comparisons of their antimicrobial resistance profiles (ARPs). When the less resistant or susceptible isolates from different genotypes were exposed to eight individual antimicrobials, they showed similar potential to become resistant with minor exceptions. This is to our knowledge the first report to examine correlations between phylogenetic sub-lineages of P. aeruginosa and their potential to become resistant to antimicrobials. This study further alerts the importance and urgency of antimicrobial abuse control.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Pseudomonas aeruginosa/classificação , Pseudomonas aeruginosa/efeitos dos fármacos , China , Cidades , Eletroforese em Gel de Campo Pulsado , Genótipo , Testes de Sensibilidade Microbiana , Tipagem Molecular , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , Estados Unidos
12.
J Basic Microbiol ; 53(7): 600-7, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22961625

RESUMO

Type VI secretion system (T6SS) has increasingly been believed to participate in the infection process for many bacterial pathogens, but its role in the virulence of Salmonella typhimurium remains unclear. To look into this, we deleted the T6SS cluster from the genome of S. typhimurium 14028s and analyzed the phenotype of the resulting T6SS knockout mutant (T6SSKO mutant) in vitro and in vivo. We found that the T6SSKO mutant exhibited reduced capability in colonizing the spleen and liver in an in vivo colonization competition model in BALB/c mice infected by the oral route. Additionally, infection via intraperitoneal administration also showed that the T6SSKO mutant was less capable of colonizing the mouse spleen and liver than the wild-type strain. We did not detect significant differences between the T6SSKO and wild-type strains in epithelial cell invasion tests. However, in the macrophage RAW264.7 cell line, the T6SSKO mutant survived and proliferated significantly more poorly than the wild-type strain. These findings indicate that T6SS gene cluster is required for full virulence of S. typhimurium 14028s in BALB/c mice, possibly due to its roles in bacterial survival and proliferation in macrophages.


Assuntos
Família Multigênica , Salmonelose Animal/microbiologia , Salmonella typhimurium/genética , Salmonella typhimurium/patogenicidade , Fatores de Virulência/genética , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Linhagem Celular , Proliferação de Células , Células HeLa , Humanos , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Salmonella typhimurium/metabolismo , Virulência/genética , Fatores de Virulência/metabolismo
13.
Front Microbiol ; 14: 1126808, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37143538

RESUMO

Many lines of evidence demonstrate the associations of colorectal cancer (CRC) with intestinal microbial dysbiosis. Recent reports have suggested that maintaining the homeostasis of microbiota and host might be beneficial to CRC patients, but the underlying mechanisms remain unclear. In this study, we established a CRC mouse model of microbial dysbiosis and evaluated the effects of fecal microbiota transplantation (FMT) on CRC progression. Azomethane and dextran sodium sulfate were used to induce CRC and microbial dysbiosis in mice. Intestinal microbes from healthy mice were transferred to CRC mice by enema. The vastly disordered gut microbiota of CRC mice was largely reversed by FMT. Intestinal microbiota from normal mice effectively suppressed cancer progression as assessed by measuring the diameter and number of cancerous foci and significantly prolonged survival of the CRC mice. In the intestine of mice that had received FMT, there were massive infiltration of immune cells, including CD8+ T and CD49b+ NK, which is able to directly kill cancer cells. Moreover, the accumulation of immunosuppressive cells, Foxp3+ Treg cells, seen in the CRC mice was much reduced after FMT. Additionally, FMT regulated the expressions of inflammatory cytokines in CRC mice, including down-regulation of IL1a, IL6, IL12a, IL12b, IL17a, and elevation of IL10. These cytokines were positively correlated with Azospirillum_sp._47_25, Clostridium_sensu_stricto_1, the E. coli complex, Akkermansia, Turicibacter, and negatively correlated with Muribaculum, Anaeroplasma, Candidatus_Arthromitus, and Candidatus Saccharimonas. Furthermore, the repressed expressions of TGFb, STAT3 and elevated expressions of TNFa, IFNg, CXCR4 together promoted the anti-cancer efficacy. Their expressions were positively correlated with Odoribacter, Lachnospiraceae-UCG-006, Desulfovibrio, and negatively correlated with Alloprevotella, Ruminococcaceae UCG-014, Ruminiclostridium, Prevotellaceae UCG-001 and Oscillibacter. Our studies indicate that FMT inhibits the development of CRC by reversing gut microbial disorder, ameliorating excessive intestinal inflammation and cooperating with anti-cancer immune responses.

14.
J Bacteriol ; 194(3): 744, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22247537

RESUMO

Salmonella enterica serovar Pullorum is a chicken-adapted pathogen, causing pullorum disease. Its strict host adaptation has been suspected to result in gene decay. To validate this hypothesis and identify the decayed genes, we sequenced the complete genome of S. Pullorum RKS5078. We found 263 pseudogenes in this strain and conducted functional analyses of the decayed genes.


Assuntos
Genoma Bacteriano , Doenças das Aves Domésticas/microbiologia , Salmonelose Animal/microbiologia , Salmonella enterica/genética , Animais , Proteínas de Bactérias/genética , Sequência de Bases , Galinhas , Dados de Sequência Molecular , Salmonella enterica/isolamento & purificação
15.
Blood Cells Mol Dis ; 48(2): 86-90, 2012 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-22197394

RESUMO

Thalassemia is the commonest inherited autosomal recessive disorders of hemoglobin in southern China. We developed and evaluated a reverse dot blot (RDB) assay combined with flow-through hybridization technology platform for the rapid and simultaneous identification of 5 types of α-thalassemia and 16 types of ß-thalassemia common in Chinese. Reliable genotyping of wild-type and thalassemic genomic DNA samples was achieved by means of a gene chip on which allele-specific oligonucleotide probes were immobilized on a nylon membrane. This method involved two multiplex PCR amplification systems of α-thalassemia and ß-thalassemia and one time of hybridization. The whole procedure starting from blood sampling to the identification of thalassemia genotype required less than 4h. The diagnostic reliability of this reverse dot blot assay was evaluated on 427 samples (387 cases of thalassemia and 40 healthy persons) by using direct DNA sequence analysis and gap-PCR in a blind study. These samples included 377 cases of blood, 7 cases of amniotic fluid, 18 cases of chorionic villus, and 25 cases of cord blood. The RDB gene chip was in complete concordance with the reference method. The reverse dot blot assay was a simple, rapid, accurate, and cost-effective method to identify common thalassemia genotypes in the Chinese population.


Assuntos
Análise de Sequência com Séries de Oligonucleotídeos/métodos , Talassemia alfa/diagnóstico , Talassemia beta/diagnóstico , Povo Asiático/genética , Sequência de Bases , China , Humanos , Reação em Cadeia da Polimerase Multiplex/métodos , Mutação , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , alfa-Globinas/genética , Talassemia alfa/genética , Globinas beta/genética , Talassemia beta/genética
16.
Zhonghua Yu Fang Yi Xue Za Zhi ; 46(4): 329-33, 2012 Apr.
Artigo em Zh | MEDLINE | ID: mdl-22800631

RESUMO

OBJECTIVE: Selecting variable-number tandem-repeat (VNTR) loci for different serogroups of Shigella spp to explore and establish multilocus variable-number tandem-repeat analysis (MLVA) method, in order to study the molecular characteristic of the isolated strains. METHODS: Of the Shigella strains found by dysentery surveillance in Beijing from 2001 to 2009, 180 strains were selected for this study, according to the number and serotypes of the surveillant strains, at the ratio of 15%; including 50 strains of Shigella sonnei and 130 strains of Shigella flexneri. After screening the polymorphism of the 18 VNTR loci, 10 VNTR loci (sh1-sh10) were retained and constructed three groups of multi-PCR methods to detect all he 180 strains and analyze MLVA molecular subtypes using capillary segments. RESULTS: A range of 2 to 11 alleles were found on the 10 VNTR loci among the 180 Shigella strains, with a diversity index value between 0.158 and 0.766. The 10 loci showed diversity in different serogroups, such as only one allele found in sh6 of Shigella flexneri, sh2 and sh3 of Shigella sonnei individually. The isolated 180 strains were divided into 84 MLVA subtypes, with a resolution ratio D value at 0.967 (95%CI: 0.956 - 0.978). The 130 strains of Shigella flexneri were divided into 63 subtypes, named as TF001-TF063; among which TF001, TF002 and TF 005 were the dominant subtypes, accounting to 17, 16 and 15 strains respectively. The 50 strains of Shigella sonnei were divided into 21 subtypes, named as TS001-TS021; among which TS002 (14 strains) and TS001 (7 strains) were the dominant subtypes. CONCLUSION: MLVA subtyping method including 10 VNTR loci was preliminarily developed. The MLVA cluster analysis revealed that the subtypes of Shigella strains isolated in Beijing were diverse, and suggested the possibility of multiple-clone source.


Assuntos
Repetições Minissatélites , Shigella/classificação , Shigella/genética , Alelos , Técnicas de Tipagem Bacteriana/métodos , China , DNA Bacteriano/genética , Genótipo , Polimorfismo Genético , Shigella/isolamento & purificação
17.
Curr Res Food Sci ; 5: 1720-1731, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36238813

RESUMO

In this study, we investigated the dynamics of microbial community and flavor metabolites during the traditional fermentation of Hongqu aromatic vinegar (HAV) and subsequently explored the potential relationship between microbiota and flavor metabolites. The microbiome analysis based on high-throughput sequencing (HTS) of amplicons demonstrated that Lactobacillus, Acetobacter and Clostridium were the dominant bacterial genera, while Alternaria, Candida, Aspergillus and Issatchenkia were the dominant fungal genera during the acetic acid fermentation (AAF) of HAV. A total of 101 volatile flavor compounds were identified through gas chromatography-mass spectrometry (GC-MS) during HAV fermentation, including esters (35), alcohols (17), aldehydes (11), acids (11), ketones (7), phenols (10), and others (10). Redundancy analysis (RDA) was used to reveal the correlation between microbiota and volatile flavor compounds. Lactobacillus and Acetobacter were the two bacterial genera that have the great influence on the production of volatile flavor components in HAV. Among them, Lactobacillus was positively correlated with a variety of ethyl esters, while Acetobacter positively contributed to the formation of several organic acids. Furthermore, the non-volatile metabolites were detected by ultra-high-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS). A total of 41 dipeptides were identified during HAV fermentation, and most of them may have sensory characteristics and biological activities. RDA showed that Aspergillus, Epicoccum, Issatchenkia, Candida and Malassezia were the most influential fungal genera on non-volatile metabolites. In particular, Epicoccum was first reported in Hongqu vinegar and showed a positive correlation with the production of various organic acids. In conclusion, this study provides a scientific basis for understanding the flavor generation mechanism of HAV, and may be valuable for developing effective techniques to select suitable strains to improve the flavor quality of HAV.

18.
Electrophoresis ; 32(5): 506-17, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21341285

RESUMO

We present a theoretical model for describing the electric field-driven migration and dispersion of short anisotropic molecules in nanofluidic filter arrays. The model uses macrotransport theory to derive exact integral-form expressions for the effective mobility and diffusivity of Brownian particles moving in an effective one-dimensional energy landscape. The latter is obtained by modeling the anisotropic molecules as point-sized Brownian particles with their orientational degrees of freedom accounted for by an entropy penalty term, and using a systematic projection procedure for reducing the system dimensionality to the device axial dimension. Our analytical results provide guidance for the design and optimization of nanofluidic separation systems without the need for complex numerical simulations. Comparison with numerical solution of the macrotransport equations in the actual, effectively two-dimensional, geometry shows that the one-dimensional model faithfully describes the field- and size-dependences of mobility and diffusivity, with maximum difference on the order of 10% under the experimentally relevant electric fields.


Assuntos
Eletroforese/métodos , Técnicas Analíticas Microfluídicas/métodos , Modelos Químicos , Anisotropia , DNA/isolamento & purificação , Filtração
19.
Zhonghua Yu Fang Yi Xue Za Zhi ; 45(9): 820-4, 2011 Sep.
Artigo em Zh | MEDLINE | ID: mdl-22177306

RESUMO

OBJECTIVE: To explore the pathogenic form, epidemic features and serotype distribution of the pathogenic bacteria causing infectious diarrhea in Beijing. METHODS: A total of 2118 samples of rectal swabs and stool specimens of diarrheal patients were collected from 6 surveillant intestinal tract clinics during the period between April and October, 2010. Enteric multiple pathogens including Vibrio cholerae, Vibrio parahaemolyticus, Salmonella, Shigella and diarrheagenic Escherichia coli were detected by the isolation culture, biochemical identification and serotyping methods. The population distribution, temporal distribution and serotype distribution of the above pathogenic bacteria were analyzed by descriptive statistical methods. RESULTS: 478 strains isolated from the total 2118 specimens were positive for pathogen detection, accounting to 22.6%. Among the 478 strains of pathogenic bacteria, Shigella accounting for 40.8% (195/478) was the most frequent pathogen, followed by Vibrio parahaemolyticus accouting for 23.8% (114/478), Salmonella accounting for 19.0% (91/478) and diarrheagenic Escherichia coli accounting for 4.8% (23/478). Enteric pathogenic bacteria spread mainly among adults aging between 20 and 39; and the distribution was different among different age groups, while the highest detected rate was in 30 - 39 age group, accounting for 27.2% (92/338). The detected rate of pathogenic bacteria showed evident seasonal variations, with a peak from July to October, whose detected rates were 23.5% (114/486), 32.8% (176/536), 36.1% (90/249) and 25.9% (29/112) respectively. The detected rates in other months were all under 16.0%. Shigella Sonnei was the dominant serotype, accounting for 83.1% (162/195). O3:K6 was the dominant serotype among Vibrio parahaemolyticus, accounting for 63.2% (72/114). Salmonella Enteritidis and Salmonella Typhimurium were dominant serotypes among Salmonella, accounting for 13.2% (12/91) and 12.1% (11/91) separately. Enterpathogenic Escherichia coli and enterotoxigenic Escherichia coli were the dominant serotypes among Diarrheagenic Escherichia coli, accounting for 69.6% (16/23) and 30.4% (7/23) respectively. CONCLUSION: The three main pathogenic bacteria causing infectious diarrhea in Beijing are Shigella, Vibrio parahaemolyticus, Salmonella; and there are obvious changes in the serotype distribution of Shigella and Samonella compared to previous years.


Assuntos
Infecções Bacterianas/epidemiologia , Diarreia/epidemiologia , Diarreia/microbiologia , Adolescente , Adulto , Idoso , Criança , China/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Salmonella/isolamento & purificação , Sorotipagem , Shigella/isolamento & purificação , Vibrio parahaemolyticus/isolamento & purificação , Adulto Jovem
20.
Zhonghua Yu Fang Yi Xue Za Zhi ; 45(2): 113-7, 2011 Feb.
Artigo em Zh | MEDLINE | ID: mdl-21426788

RESUMO

OBJECTIVE: To study the epidemiological characteristics and molecular phenotypes of Salmonella by pulsed-field gel electrophoresis (PFGE) in Beijing from 2008 to 2009. METHODS: A total of one hundred thirty-seven isolates recovered from the WHO Global Salmonella Surveillance system and entero clinic surveillance system were identified by biochemical tests and serotyping. The related epidemiological informations were also analyzed. The isolates were further typed by PFGE. RESULTS: The prevalence of Salmonella from 2008 to 2009 showed obvious seasonal character. High incidence occurred from June to September, and 64.1% (84/131) isolates were recovered in this period. Patients of 18 - 40 year-old were 46.1% (58/128) and 80 patients were male and 40 patients were female with the ratio of 1.57:1. These 137 Salmonella isolates belonged to 20 serotypes, including Enteritidis (46.7%, 64/137) and Typhimurium (17.5%, 24/137) as the dominant serotype. In total, 71 PFGE profiles were identified. Four PFGE patterns of S. Enteritidis isolates (JEGX01.CN0001, JEGX01.CN0003, JEGX01.CN0002, JEGX01.CN0019) and S. Typhimurium pattern of JPXX01.CN0001 were dominant patterns. CONCLUSION: The prevalence of Salmonella from 2008 to 2009 showed distribution characteristics of sex, age and seasons. The numerous PFGE patterns of Salmonella showed diversity of these isolates and different clones existed in Beijing.


Assuntos
Tipagem Molecular , Infecções por Salmonella/microbiologia , Salmonella/isolamento & purificação , Adolescente , Adulto , Técnicas de Tipagem Bacteriana , China/epidemiologia , DNA Bacteriano/isolamento & purificação , Eletroforese em Gel de Campo Pulsado , Feminino , Microbiologia de Alimentos , Humanos , Masculino , Salmonella/classificação , Salmonella/genética , Infecções por Salmonella/epidemiologia , Sorotipagem , Adulto Jovem
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