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1.
Biochim Biophys Acta Biomembr ; 1864(4): 183866, 2022 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-35007517

RESUMO

In light of an increasing number of antibiotic-resistant bacterial strains, it is essential to understand an action imposed by various antimicrobial agents on bacteria at the molecular level. One of the leading mechanisms of killing bacteria is related to the alteration of their plasmatic membrane. We study bio-inspired peptides originating from natural antimicrobial proteins colicins, which can disrupt membranes of bacterial cells. Namely, we focus on the α-helix H1 of colicin U, produced by bacterium Shigella boydii, and compare it with analogous peptides derived from two different colicins. To address the behavior of the peptides in biological membranes, we employ a combination of molecular simulations and experiments. We use molecular dynamics simulations to show that all three peptides are stable in model zwitterionic and negatively charged phospholipid membranes. At the molecular level, their embedment leads to the formation of membrane defects, membrane permeation for water, and, for negatively charged lipids, membrane poration. These effects are caused by the presence of polar moieties in the considered peptides. Importantly, simulations demonstrate that even monomeric H1 peptides can form toroidal pores. At the macroscopic level, we employ experimental co-sedimentation and fluorescence leakage assays. We show that the H1 peptide of colicin U incorporates into phospholipid vesicles and disrupts their membranes, causing leakage, in agreement with the molecular simulations. These insights obtained for model systems seem important for understanding the mechanisms of antimicrobial action of natural bacteriocins and for future exploration of small bio-inspired peptides able to disrupt bacterial membranes.


Assuntos
Colicinas/metabolismo , Fosfolipídeos/química , Lipossomas Unilamelares/metabolismo , Sequência de Aminoácidos , Colicinas/química , Colicinas/farmacologia , Simulação de Dinâmica Molecular , Permeabilidade/efeitos dos fármacos , Fosfatidilcolinas/química , Fosfatidiletanolaminas/química , Conformação Proteica em alfa-Hélice , Shigella boydii/metabolismo , Lipossomas Unilamelares/química
2.
Carbohydr Polym ; 233: 115842, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32059894

RESUMO

This work discussed the preparation of Ag nanoparticles (AgNPs) and AgNPs impregnated sub-micrometer crystalline jute cellulose (SCJC) particles using a green synthetic bioreduction method. The ultimate nanocomposite particles were named as SCJC/Ag. The crystalline structure of AgNPs was maintained in SCJC/Ag nanocomposte particles. The catalytic efficiency of SCJC/Ag nanocomposite particles were evaluated for the degradation of congo red (CR) and methylene blue (MB) using NaBH4 as reducing agent. A complete degradation of 20 mL of each CR (0.1 mM) and MB (0.05 mM) dye solution was achieved within 14 min when 0.005 mg mL-1 of SCJC/Ag nanocomposite particles was employed. SCJC/Ag nanocomposite particles also exhibited moderate antibacterial activities against Staphylococcus aureus, Escherichia coli, Shigella dysenteriae and Shigella boydii and the results were comparable with those of the reference AgNPs. SCJC/Ag nanocomposite particles were the most effective against Escherichia coli (E. coli) with minimum inhibitory concentration of 0.014 mg mL-1.


Assuntos
Antibacterianos/farmacologia , Celulose/química , Corchorus/química , Nanopartículas Metálicas/química , Prata/farmacologia , Boroidretos/química , Catálise , Vermelho Congo/química , Escherichia coli/efeitos dos fármacos , Azul de Metileno/química , Testes de Sensibilidade Microbiana , Oxirredução , Rubiaceae/química , Shigella boydii/efeitos dos fármacos , Shigella dysenteriae/efeitos dos fármacos , Prata/química , Staphylococcus aureus/efeitos dos fármacos
3.
J Bacteriol ; 201(24)2019 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-31548276

RESUMO

Colicin U is a protein produced by the bacterium Shigella boydii (serovars 1 and 8). It exerts antibacterial activity against strains of the enterobacterial genera Shigella and Escherichia Here, we report that colicin U forms voltage-dependent pores in planar lipid membranes; its single-pore conductance was found to be about 22 pS in 1 M KCl at pH 6 under 80 mV in asolectin bilayers. In agreement with the high degree of homology between their C-terminal domains, colicin U shares some pore characteristics with the related colicins A and B. Colicin U pores are strongly pH dependent, and as we deduced from the activity of colicin U in planar membranes at different protein concentrations, they have a monomeric pore structure. However, in contrast to related colicins, we observed a very low cationic selectivity of colicin U pores (1.5/1 of K+/Cl- at pH 6) along with their atypical voltage gating. Finally, using nonelectrolytes, we determined the inner diameter of the pores to be in the range of 0.7 to 1 nm, which is similar to colicin Ia, but with a considerably different inner profile.IMPORTANCE Currently, a dramatic increase in antibiotic resistance is driving researchers to find new antimicrobial agents. The large group of toxins called bacteriocins appears to be very promising from this point of view, especially because their narrow killing spectrum allows specific targeting against selected bacterial strains. Colicins are a subgroup of bacteriocins that act on Gram-negative bacteria. To date, some colicins are commercially used for the treatment of animals (1) and tested as a component of engineered species-specific antimicrobial peptides, which are studied for the potential treatment of humans (2). Here, we present a thorough single-molecule study of colicin U which leads to a better understanding of its mode of action. It extends the range of characterized colicins available for possible future medical applications.


Assuntos
Membrana Celular/metabolismo , Colicinas/metabolismo , Bicamadas Lipídicas/metabolismo , Shigella boydii/metabolismo , Concentração de Íons de Hidrogênio , Ativação do Canal Iônico , Permeabilidade , Cloreto de Potássio/farmacologia
4.
J Biochem Mol Toxicol ; 33(4): e22281, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30597695

RESUMO

A series of 1,3-bis-chalcone derivatives (3a-i, 6a-i and 8) were synthesized and evaluated antimicrobial, antibiofilm and carbonic anhydrase inhibition activities. In this evaluation, 6f was found to be the most active compound showing the same effect as the positive control against Bacillus subtilis and Streptococcus pyogenes in terms of antimicrobial activity. Biofilm structures formed by microorganisms were damaged by compounds at the minimum inhibitory concentration value between 0.5% and 97%.1,3-bis-chalcones ( 3a-i, 6a-i and 8) showed good inhibitory action against human (h) carbonic anhydrase (CA) isoforms I and II. hCA I and II were effectively inhibited by these compounds, with K i values in the range of 94.33 ± 13.26 to 787.38 ± 82.64 nM for hCA I, and of 100.37 ± 11.41 to 801.76 ± 91.11 nM for hCA II, respectively. In contrast, acetazolamide clinically used as CA inhibitor showed K i value of 1054.38 ± 207.33 nM against hCA I, and 983.78 ± 251.08 nM against hCA II, respectively.


Assuntos
Inibidores da Anidrase Carbônica/farmacologia , Chalconas/farmacologia , Isoenzimas/antagonistas & inibidores , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Bacillus subtilis/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Inibidores da Anidrase Carbônica/química , Chalconas/química , Avaliação Pré-Clínica de Medicamentos , Humanos , Testes de Sensibilidade Microbiana , Espectroscopia de Prótons por Ressonância Magnética , Pseudomonas aeruginosa/efeitos dos fármacos , Shigella boydii/efeitos dos fármacos , Streptococcus pyogenes/efeitos dos fármacos
5.
Int J Biol Macromol ; 124: 819-827, 2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-30496858

RESUMO

We studied localization and physiological activities of a lectin showing specific binding to N-acetylhexosamines, termed HOL-18, purified from Japanese black sponge (Halichondria okadai). Antiserum against the lectin was generated in rabbit and applied for immunohistochemical analyses. HOL-18 was expressed specifically around water pores and on spicules of sponge tissues. It showed strong binding to a variety of N-acetylhexosamines: N-acetyl D-glucosamine, N-acetyl D-galactosamine, N-acetyl mannosamine, N-acetyl muramic acid, and N-acetyl neuraminic acid. Hemagglutination induced by the lectin was inhibited by lipopolysaccharides and a peptidoglycan. HOL-18 inhibited growth of a gram-positive bacterium (Listeria monocytogenes), gram-negative bacteria (Escherichia coli, Shigella boydii, Pseudomonas aeruginosa), and a fungus (Aspergillus niger). It displayed anti-biofilm activity against P. aeruginosa. HOL-18 was internalized into conidiophores of A. niger, and displayed notable antifungal activity. Fluorescence microscopy revealed binding and incorporation of the lectin into human cancer cell lines HeLa, MCF-7, and T47D, but not Caco-2. HOL-18 displayed dose-dependent cytotoxic effects against HeLa, MCF-7, and T47D, with respective IC50 values 40, 52, and 63 µg/mL. In HeLa cells, it activated phosphorylation of MAPK pathway molecule (ERK1/2) and activated caspase-3 to trigger apoptosis. HOL-18 thus has the potential to upregulate metabolic pathways in higher animal cells through binding to N-acetylhexosamines.


Assuntos
Anti-Infecciosos/química , Antineoplásicos/química , Hexosaminas/química , Lectinas/química , Poríferos/química , Animais , Anti-Infecciosos/isolamento & purificação , Anti-Infecciosos/metabolismo , Anti-Infecciosos/farmacologia , Antineoplásicos/isolamento & purificação , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Aspergillus niger/efeitos dos fármacos , Aspergillus niger/crescimento & desenvolvimento , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Caspase 3/genética , Caspase 3/metabolismo , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Células HeLa , Testes de Hemaglutinação , Hexosaminas/metabolismo , Humanos , Lectinas/isolamento & purificação , Lectinas/metabolismo , Lectinas/farmacologia , Lipopolissacarídeos/farmacologia , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/crescimento & desenvolvimento , Células MCF-7 , Testes de Sensibilidade Microbiana , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Peptidoglicano/farmacologia , Fosforilação/efeitos dos fármacos , Ligação Proteica , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/crescimento & desenvolvimento , Coelhos , Shigella boydii/efeitos dos fármacos , Shigella boydii/crescimento & desenvolvimento
6.
J Med Microbiol ; 67(11): 1596-1600, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30251950

RESUMO

In January 2015, Public Health England and the United Kingdom (UK) Ministry of Defence investigated cases of diarrhoea and fever in military personnel recently returned to the UK after supporting the response to the Ebola epidemic in Sierra Leone. Tests for Ebola virus infection were negative. PCR tests detected the ipaH gene in 10/12 faecal specimens, and Shigella boydii serotype 20 was isolated from 7 patients. A case control study was undertaken and analysed using multivariable logistic regression. Consumption of a coronation chicken lunch at the transit camp in Sierra Leone (SL) 24-48 h prior to departure for the UK was significantly associated with disease [adjusted odds ratio (OR) 28.15, 95 % CI: 1.87-422.65]. In the context of heightened concern during the Ebola epidemic, this outbreak highlights the importance of rapid and effective microbiological and epidemiological investigations to identify the aetiological agent in patients presenting with fever and diarrhoea.


Assuntos
Doenças Transmissíveis Importadas/microbiologia , Surtos de Doenças , Disenteria Bacilar/epidemiologia , Pessoal de Saúde , Doença pelo Vírus Ebola/epidemiologia , Shigella boydii/isolamento & purificação , Adulto , Animais , Proteínas de Bactérias/genética , Estudos de Casos e Controles , Galinhas/microbiologia , Doenças Transmissíveis Importadas/epidemiologia , Disenteria Bacilar/etiologia , Disenteria Bacilar/microbiologia , Fezes/microbiologia , Feminino , Febre/epidemiologia , Febre/microbiologia , Contaminação de Alimentos , Doença pelo Vírus Ebola/virologia , Humanos , Cooperação Internacional , Masculino , Militares , Sorogrupo , Shigella boydii/classificação , Shigella boydii/genética , Shigella boydii/imunologia , Serra Leoa/epidemiologia , Reino Unido/epidemiologia , Sequenciamento Completo do Genoma , Adulto Jovem
7.
Epidemiol Infect ; 146(11): 1445-1451, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29880081

RESUMO

Shigellosis causes significant morbidity and mortality in developing and developed countries, mostly among infants and young children. The World Health Organization estimates that more than one million people die from Shigellosis every year. In order to evaluate trends in Shigellosis in Israel in the years 2002-2015, we analysed national notifiable disease reporting data. Shigella sonnei was the most commonly identified Shigella species in Israel. Hospitalisation rates due to Shigella flexenri were higher in comparison with other Shigella species. Shigella morbidity was higher among infants and young children (age 0-5 years old). Incidence of Shigella species differed among various ethnic groups, with significantly high rates of S. flexenri among Muslims, in comparison with Jews, Druze and Christians. In order to improve the current Shigellosis clinical diagnosis, we developed machine learning algorithms to predict the Shigella species and whether a patient will be hospitalised or not, based on available demographic and clinical data. The algorithms' performances yielded an accuracy of 93.2% (Shigella species) and 94.9% (hospitalisation) and may consequently improve the diagnosis and treatment of the disease.


Assuntos
Algoritmos , Disenteria Bacilar/epidemiologia , Shigella boydii , Shigella dysenteriae , Shigella flexneri , Shigella sonnei , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Cristianismo , Disenteria Bacilar/etnologia , Disenteria Bacilar/microbiologia , Disenteria Bacilar/mortalidade , Feminino , Hospitalização , Humanos , Incidência , Lactente , Islamismo , Israel/epidemiologia , Judeus , Modelos Logísticos , Aprendizado de Máquina , Masculino , Pessoa de Meia-Idade , Modelos Estatísticos , Redes Neurais de Computação , Reconhecimento Automatizado de Padrão , Adulto Jovem
8.
Eur J Med Chem ; 128: 274-286, 2017 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-28192710

RESUMO

In some organisms, environmental stress triggers trehalose biosynthesis that is catalyzed collectively by trehalose 6-phosphate synthase, and trehalose 6-phosphate phosphatase (T6PP). T6PP catalyzes the hydrolysis of trehalose 6-phosphate (T6P) to trehalose and inorganic phosphate and is a promising target for the development of antibacterial, antifungal and antihelminthic therapeutics. Herein, we report the design, synthesis and evaluation of a library of aryl d-glucopyranoside 6-sulfates to serve as prototypes for small molecule T6PP inhibitors. Steady-state kinetic techniques were used to measure inhibition constants (Ki) of a panel of structurally diverse T6PP orthologs derived from the pathogens Brugia malayi, Ascaris suum, Mycobacterium tuberculosis, Shigella boydii and Salmonella typhimurium. The binding affinities of the most active inhibitor of these T6PP orthologs, 4-n-octylphenyl α-d-glucopyranoside 6-sulfate (9a), were found to be in the low micromolar range. The Ki of 9a with the B. malayi T6PP ortholog is 5.3 ± 0.6 µM, 70-fold smaller than the substrate Michaelis constant. The binding specificity of 9a was demonstrated using several representative sugar phosphate phosphatases from the HAD enzyme superfamily, the T6PP protein fold family of origin. Lastly, correlations drawn between T6PP active site structure, inhibitor structure and inhibitor binding affinity suggest that the aryl d-glucopyranoside 6-sulfate prototypes will find future applications as a platform for development of tailored second-generation T6PP inhibitors.


Assuntos
Desenho de Fármacos , Inibidores Enzimáticos/farmacologia , Glucosiltransferases/antagonistas & inibidores , Monossacarídeos/farmacologia , Fosfatos Açúcares/metabolismo , Trealose/análogos & derivados , Animais , Brugia Malayi/enzimologia , Inibidores Enzimáticos/química , Ensaios de Triagem em Larga Escala , Humanos , Mycobacterium tuberculosis/enzimologia , Salmonella typhimurium/enzimologia , Shigella boydii/enzimologia , Trealose/metabolismo
9.
Artigo em Inglês | MEDLINE | ID: mdl-27148494

RESUMO

Shigella is a pathovar of Escherichia coli comprising four groups, Shigella flexneri, Shigella sonnei, Shigella dysenteriae, and Shigella boydii, each of them, with the exception of S.sonnei, comprising several serotypes. Shigella accounts for the majority of dysentery causing infections occurring world-wide each year. Recent advancements in the Shigella field have led to a better understanding of the molecular mechanisms underlying host epithelial cell invasion and immune cell function manipulation, mainly using S. flexneri as a model. Host-cell invasion is the final step of the infection process, as Shigella's virulence strategy relies also on its ability to survive hostile conditions during its journey through the gastro-intestinal tract, to compete with the host microbiota and to cross the intestinal mucus layer. Hence, the diversity of the virulence strategies among the different Shigella species has not yet been deeply investigated, which might be an important step to understand the epidemiological spreading of Shigella species worldwide and a key aspect for the validation of novel vaccine candidates. The recent development of high-throughput screening and sequencing methods will facilitate these complex comparison studies. In this review we discuss several of the major avenues that the Shigella research field has taken over the past few years and hopefully gain some insights into the questions that remain surrounding this important human pathogen.


Assuntos
Disenteria Bacilar/epidemiologia , Shigella boydii/patogenicidade , Shigella dysenteriae/patogenicidade , Shigella flexneri/patogenicidade , Shigella sonnei/patogenicidade , Disenteria Bacilar/imunologia , Disenteria Bacilar/microbiologia , Células Epiteliais/microbiologia , Geografia , Interações Hospedeiro-Patógeno , Humanos
10.
Pathog Dis ; 74(4): ftw027, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27056949

RESUMO

ITALIC! Shigella boydiiis one of the four ITALIC! Shigellaspecies that causes disease worldwide; however, there are few published studies that examine the genomic variation of this species. This study compares genomes of 72 total isolates; 28 ITALIC! S. boydiifrom Bangladesh and The Gambia that were recently isolated as part of the Global Enteric Multicenter Study (GEMS), 14 historical ITALIC! S. boydiigenomes in the public domain and 30 ITALIC! Escherichia coliand ITALIC! Shigellareference genomes that represent the genomic diversity of these pathogens. This comparative analysis of these 72 genomes identified that the ITALIC! S. boydiiisolates separate into three phylogenomic clades, each with specific gene content. Each of the clades contains ITALIC! S. boydiiisolates from geographic and temporally distant sources, indicating that the ITALIC! S. boydiiisolates from the GEMS are representative of ITALIC! S. boydii.This study describes the genome sequences of a collection of novel ITALIC! S. boydiiisolates and provides insight into the diversity of this species in comparison to the ITALIC! E. coliand other ITALIC! Shigellaspecies.


Assuntos
Disenteria Bacilar/microbiologia , Variação Genética , Genoma Bacteriano , Shigella boydii/classificação , Shigella boydii/genética , Biologia Computacional/métodos , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Filogenia
12.
Vaccine ; 34(15): 1839-46, 2016 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-26878295

RESUMO

Our previous studies on outer membrane vesicles based vaccine development against shigellosis, revealed the inability of Shigella to release significant amount of vesicles naturally, during growth. Disruption of tolA, one of the genes of the Tol-Pal system of Gram negative bacterial membrane, has increased the vesicle release rate of a Shigella boydii type 4 strain to approximately 60% higher. We also noticed the vesicles, released from tolA-disrupted strain captured more OmpA protein and lipopolysaccharide, compared to the vesicles released from its wild type prototype. Six to seven weeks old BALB/c mice, immunized with 25 µg of three oral doses of the vesicles, released by tolA mutant, conferred 100% protection against lethal homologous challenge through nasal route, compared to only 60% protection after the same dose of wild type immunogen. Mice, immunized with the vesicles from tolA-mutant, manifested significant secretion of mucosal IgG and IgA. A sharp and significant response of pro-inflammatory cytokines (TNF-α, IL-6, IFN-γ) were also observed in the lung lavage of these groups of mice, within 6h post challenge; but at 24h, these inflammatory cytokines showed the sign of subsidence and the system was taken over by the release of anti-inflammatory cytokines (IL-4 and IL-10). Studies with naïve peritoneal macrophages, proved further, the potency of these vesicles to stimulate nitric oxide and TNF-α, IL-12p70, IL-6 and IL-10 productions in-vitro. The ability of these vesicles to trigger polarization of CD4(+) T cells toward Th1 adaptive immune response, had also been observed along with the presence of anti-inflammatory cytokines in the system. Our study demonstrated, the vesicles from tolA-disrupted Shigella were able to suppress Shigella-mediated inflammation in the host and could balance between inflammation and anti-inflammation, promoting better survival and health of the infected mice. Outer membrane vesicles from tolA-mutant, could be a potential cost-effective vaccine candidate against shigellosis.


Assuntos
Vesículas Extracelulares/imunologia , Vacinas contra Shigella/imunologia , Shigella boydii , Administração Oral , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/química , Proteínas da Membrana Bacteriana Externa/imunologia , Líquido da Lavagem Broncoalveolar/química , Citocinas/química , Disenteria Bacilar/prevenção & controle , Técnicas de Inativação de Genes , Pulmão/imunologia , Macrófagos Peritoneais/imunologia , Camundongos Endogâmicos BALB C , Shigella boydii/genética
13.
Carbohydr Res ; 414: 46-50, 2015 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-26232763

RESUMO

The O-polysaccharide (O-antigen) of Escherichia coli O169 was studied by sugar analysis along with 1D and 2D (1)H and (13)C NMR spectroscopy. The following structure of the branched hexasaccharide repeating unit was established: [Formula: see text] The O-polysaccharide of E. coli O169 differs from that of Shigella boydii type 6 only in the presence of a side-chain glucose residue. A comparison of the O-antigen biosynthesis gene clusters between the galF to gnd genes in the genomes of the two bacteria revealed their close relationship. The glycosyltransferase gene responsible for the formation of the ß-D-Glcp-(1 → 6)-α-D-Galp linkage in the O-antigen was identified in the gene cluster.


Assuntos
Escherichia coli/genética , Antígenos O/química , Antígenos O/genética , Shigella boydii/genética , Sequência de Carboidratos , Escherichia coli/metabolismo , Genoma Bacteriano , Espectroscopia de Ressonância Magnética , Família Multigênica , Shigella boydii/metabolismo
14.
Analyst ; 140(19): 6595-601, 2015 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-26300105

RESUMO

Being high throughput, rapid, automated, economical, convenient to operate and highly sensitive, protein arrays have been widely used in the analysis of tumor markers and veterinary drug residues. Pathogenic microbes also can be detected qualitatively by DNA array or protein array; however, their high throughput detection and quantification remains a big obstacle. To evaluate the potentiality of protein arrays for multiple quantitative detection of microorganisms with naked eye examination without the help of any equipment, here we developed a visual-antibody-macroarray (VAMA) aiming at rapid and simultaneous quantification of Escherichia coli O157:H7 and Shigella boydii. The results show that this VAMA is highly specific and is able to distinguish mixed Escherichia coli O157:H7 and Shigella boydii synchronously. The detection limits are equivalent to 3.4 × 10(5) CFU mL(-1) and 3.2 × 10(5) CFU mL(-1), respectively, which conform to the results of plate counting and ELISA. Importantly, the examination can be solely performed with the naked eye. Therefore, we provide an easy, reliable and rapid method for quantitative analysis of microorganisms.


Assuntos
Anticorpos Monoclonais/imunologia , Escherichia coli O157/isolamento & purificação , Análise Serial de Proteínas/métodos , Shigella boydii/isolamento & purificação , Colódio/química , Escherichia coli O157/imunologia , Limite de Detecção , Membranas Artificiais , Shigella boydii/imunologia , Fatores de Tempo
15.
Enferm. infecc. microbiol. clín. (Ed. impr.) ; 33(6): 379-384, jun.-jul. 2015. ilus
Artigo em Espanhol | IBECS | ID: ibc-142119

RESUMO

INTRODUCCIÓN: El objetivo de este estudio fue describir la evolución y las características epidemiológicas de los pacientes con shigelosis durante 25 años en una gran ciudad. MÉTODOS: La shigelosis es una enfermedad de declaración obligatoria en España desde 1988. Se analizan los casos de residentes en Barcelona incluidos en el registro entre 1988-2012. Se presenta un análisis descriptivo según sexo, edad, vía de transmisión y especies de Shigella. Se realizó un análisis de tendencias y de series temporales. RESULTADOS: De los 559 casos analizados, el 60,15% correspondían a hombres. Se observó un incremento sostenido de la tendencia en hombres desde 2008 (p < 0,05), sobre todo a expensas de los de hombres que no tenían antecedentes de toXIInfección alimentaria ni de viajes a zonas endémicas. El incremento de la tendencia fue mayor en hombres de 21 a 60 años, tanto para S. flexneri (desde 2009) como para S. sonnei (desde 2003). En 2012 se observó que, en los hombres con S. flexneri, el 63% tenían sexo con hombres. CONCLUSIONES: Se detectó un incremento de la tendencia en los casos en hombres que no tenían antecedentes de toXIInfección alimentaria ni de viajes a zonas endémicas. Este incremento apunta a un cambio en el patrón de la shigelosis, pasando a ser predominantemente masculina, y cuyo mecanismo principal serían las relaciones sexuales


INTRODUCTION: The aim of the study was to analyze the incidence, management and cost associated to hematological and dermatological adverse effects (AE) in chronic hepatitis C patients on triple therapy (TT) with telaprevir (TVR) or boceprevir (BOC). METHODS: An analysis was made on the data recorded on patients who started treatment with TVR or BOC associated with peginterferon alfa and ribavirin in a 12-week follow-up period. RESULTS: Fifty-three patients were included (TVR n = 36; BOC n = 17). Thrombocytopenia (83% TVR vs. 88% BOC) followed by neutropenia (89% TVR vs. 82% BOC) were the most common AE. Dermatological AE were observed in 32% of patients. Eleven patients required treatment discontinuation (all of them received TVR), and toxicity was the main reason for discontinuation (64%). The percentage of patients who required supportive treatment for management of AE was 66%. The most used supportive treatment was erythropoietin. Eight patients required emergency health care, and 2 were hospitalized due to AE. Total cost of additional supportive resources was 32,522 Euros (625 [SD = 876] Euros/patient) (TVR 759 [SD = 1,022] Euros/patient vs. BOC 349 [SD = 327] Euros/patient; P > .05). Patients with grade iii-iv toxicity required greater supportive care with higher costs, compared to patients with grade i-ii toxicity (849 [SD = 1,143] Euros/patient vs. 387 [SD = 397] Euros/patient; P = .053). CONCLUSION: The addition of new protease inhibitors to conventional treatment leads to a higher incidence of hematological AE in our study, compared to data described in clinical trials. The elevated incidence of AE involves the use of supportive care, increasing total costs of therapy


Assuntos
Adulto , Feminino , Humanos , Masculino , Disenteria Bacilar/epidemiologia , Disenteria Bacilar/mortalidade , Disenteria Bacilar/transmissão , Doenças Transmitidas por Alimentos/diagnóstico , Shigella boydii/patogenicidade , Shigella dysenteriae/patogenicidade , Shigella flexneri/patogenicidade , Monitoramento Epidemiológico/tendências , Notificação de Doenças , Homossexualidade Masculina , Comportamento Sexual , Infecções Sexualmente Transmissíveis , Surtos de Doenças , Saúde do Viajante , Espanha/epidemiologia
16.
Pak J Pharm Sci ; 28(3): 833-40, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-26004715

RESUMO

The present research work investigates the in vitro antimicrobial activity of different solvent extracted samples from the aerial parts (stem, leaf, fruit and flower) of C. roseus against different microbial species using disc diffusion assay at two different concentrations of 1 and 2 mg disc-1. Hexane extracted samples inhibited the growth of all tested microbial strains except S. typhi. Similarly, ethyl acetate extracted samples was effective to control the activity of all the tested microbial strains. E. coli and S. typhi showed resistance to chloroform extracted samples and the remaining eight microbial strains were susceptible to the same extract. Butanol extracted samples did not inhibit the growth of K. pneumonia and S. typhi at low concentration, however, at higher concentration the same extract reduced the growth of different microbes. Methanol extracted samples effectively controlled the growth of all tested microbes at both concentrations except for S. typhi. Water extracted samples did not inhibit the growth at low concentration except E. coli, K. pneumonia and S. aureus and were ineffective against P. aeroginosa at both concentration. C. albicans, showed resistance against chloroform and water extracted samples at low concentration and susceptible to other solvent extracted samples at both concentration. All fractions were effective against plant pathogens i.e. E. carotovora and A. tumefaciens.


Assuntos
Anti-Infecciosos/farmacologia , Catharanthus , Componentes Aéreos da Planta , Extratos Vegetais/farmacologia , Agrobacterium tumefaciens/efeitos dos fármacos , Bacillus subtilis/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Erwinia/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Klebsiella pneumoniae/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa , Salmonella typhi/efeitos dos fármacos , Shigella boydii/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos
17.
Res Microbiol ; 165(8): 671-8, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25283727

RESUMO

Shigellosis is one of major foodborne pathogens in both developed and developing countries. Although antibiotic therapy is considered an effective treatment for shigellosis, the imprudent use of antibiotics has led to the increase of multiple-antibiotic-resistant Shigella species globally. In this study, we isolated a virulent Podoviridae bacteriophage (phage), pSb-1, that infects Shigella boydii. One-step growth analysis revealed that this phage has a short latent period (15 min) and a large burst size (152.63 PFU/cell), indicating that pSb-1 has good host infectivity and effective lytic activity. The double-stranded DNA genome of pSb-1 is composed of 71,629 bp with a G + C content of 42.74%. The genome encodes 103 putative ORFs, 9 putative promoters, 21 transcriptional terminators, and one tRNA region. Genome sequence analysis of pSb-1 and comparative analysis with the homologous phage EC1-UPM, N4-like phage revealed that there is a high degree of similarity (94%, nucleotide sequence identity) between pSb-1 and EC1-UPM in 73 of the 103 ORFs of pSb-1. The results of this investigation indicate that pSb-1 is a novel virulent N4-like phage infecting S. boydii and that this phage might have potential uses against shigellosis.


Assuntos
Bacteriófagos/genética , Bacteriófagos/isolamento & purificação , DNA Viral/química , DNA Viral/genética , Genoma Viral , Shigella boydii/virologia , Bacteriófagos/crescimento & desenvolvimento , Composição de Bases , Vírus de DNA/genética , Vírus de DNA/isolamento & purificação , Genes Virais , Anotação de Sequência Molecular , Dados de Sequência Molecular , Podoviridae/genética , Podoviridae/isolamento & purificação , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Sintenia
18.
J Med Microbiol ; 63(Pt 12): 1696-1703, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25261061

RESUMO

During 2000-2004, 13 Shigella strains that were untypable by commercially available antisera were isolated from children <5 years of age with acute diarrhoea in Kolkata. These strains were subsequently identified as Shigella dysenteriae provisional serovar 204/96 (n = 3), Shigella dysenteriae provisional serovar E23507 (n = 1), Shigella dysenteriae provisional serovar I9809-73 (n = 1), Shigella dysenteriae provisional serovar 93-119 (n = 1), Shigella flexneri provisional serovar 88-893 (n = 6) and Shigella boydii provisional serovar E16553 (n = 1). In this study, characterization of those provisional serovars of Shigella was performed with respect to their antimicrobial resistance, plasmids, virulence genes and PFGE profiles. The drug resistant strains (n = 10) of Shigella identified in this study possessed various antibiotic resistance genetic markers like catA (for chloramphenicol resistance); tetA and tetB (for tetracycline resistance); dfrA1 and sul2 (for co-trimoxazole resistance); aadA1, strA and strB (for streptomycin resistance) and blaOXA-1 (for ampicillin resistance). Class 1 and/or class 2 integrons were present in eight resistant strains. Three study strains were pan-susceptible. A single mutation in the gyrA gene (serine to leucine at codon 83) was present in four quinolone resistant strains. The virulence gene ipaH (invasion plasmid antigen H) was uniformly present in all strains in this study, but the stx (Shiga toxin) and set1 (Shigella enterotoxin 1) genes were absent. Other virulence genes like ial (invasion associated locus) and sen (Shigella enterotoxin 2) were occasionally present. A large plasmid of 212 kb and of incompatibility type IncFIIA was present in the majority of the strains (n = 10) and diversity was noticed in the smaller plasmid profiles of these strains even within the same provisional serovars. PFGE profile analysis showed the presence of multiple unrelated clones among the isolates of provisional Shigella serovars. To the best of our knowledge, this is the first report on the phenotypic and molecular characterization of provisional serovars of Shigella isolates from Kolkata, India.


Assuntos
Disenteria Bacilar/microbiologia , Tipagem Molecular , Shigella boydii/isolamento & purificação , Shigella dysenteriae/isolamento & purificação , Shigella flexneri/isolamento & purificação , DNA Bacteriano/genética , Farmacorresistência Bacteriana , Eletroforese em Gel de Campo Pulsado , Genes Bacterianos , Humanos , Índia , Plasmídeos/análise , Sorogrupo , Shigella boydii/classificação , Shigella boydii/genética , Shigella boydii/imunologia , Shigella dysenteriae/classificação , Shigella dysenteriae/genética , Shigella dysenteriae/imunologia , Shigella flexneri/classificação , Shigella flexneri/genética , Shigella flexneri/imunologia , Fatores de Virulência/genética
19.
J Bacteriol ; 196(11): 2030-42, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24659774

RESUMO

We report the development of SearchDOGS Bacteria, software to automatically detect missing genes in annotated bacterial genomes by combining BLAST searches with comparative genomics. Having successfully applied the approach to yeast genomes, we redeveloped SearchDOGS to function as a standalone, downloadable package, requiring only a set of GenBank annotation files as input. The software automatically generates a homology structure using reciprocal BLAST and a synteny-based method; this is followed by a scan of the entire genome of each species for unannotated genes. Results are provided in a HTML interface, providing coordinates, BLAST results, syntenic location, omega values (Ka/Ks, where Ks is the number of synonymous substitutions per synonymous site and Ka is the number of nonsynonymous substitutions per nonsynonymous site) for protein conservation estimates, and other information for each candidate gene. Using SearchDOGS Bacteria, we identified 155 gene candidates in the Shigella boydii sb227 genome, including 56 candidates of length < 60 codons. SearchDOGS Bacteria has two major advantages over currently available annotation software. First, it outperforms current methods in terms of sensitivity and is highly effective at identifying small or highly diverged genes. Second, as a freely downloadable package, it can be used with unpublished or confidential data.


Assuntos
Bases de Dados Genéticas , Genoma Bacteriano , Software , Sequência de Bases , Genômica , Homologia de Sequência do Ácido Nucleico , Shigella boydii/genética , Sintenia
20.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-110396

RESUMO

Shigella bacteremia is rare, occurring mainly in children. Shigella species often cause diarrhea or gastrointestinal inflammation in humans and are rarely associated with bacteremia. This report describes an unusual case of Shigella boydii bacteremia in an 84-year-old patient visiting our hospital after experiencing nausea, vomiting, and febrile sensation for 2 days. Peripheral blood cultures revealed S. boydii and 16S rDNA sequence analysis produced the same result. However, the organism was not isolated from the patient's stool. She was started on ciprofloxacin, to which this organism is sensitive, and was subsequently discharged with instructions to complete a 14-day course of ciprofloxacin. Shigellosis is usually a self-limiting enteric disease. However, in contrast to its isolation from both blood and stool, isolation of the organism from blood only is associated with a high mortality rate. As is frequently pointed out, blood cultures should be obtained from elderly or immunocompromised patients with acute febrile gastroenteritis to detect infection caused by enteric pathogens, including Shigella.


Assuntos
Idoso , Idoso de 80 Anos ou mais , Criança , Humanos , Bacteriemia , Ciprofloxacina , Diarreia , DNA Ribossômico , Disenteria Bacilar , Gastroenterite , Hospedeiro Imunocomprometido , Inflamação , Mortalidade , Náusea , Sensação , Análise de Sequência , Shigella boydii , Shigella , Vômito
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