RESUMO
BACKGROUND: Pseudomonas aeruginosa produces the Sec and Tat protein secretion machineries. The latter appears to be involved in the secretion of virulence factors, including phospholipase C (PlcH), and hence is a potential target of chemotherapeutic agents. METHODS: The signal sequence of OprM, the outer membrane subunit of the xenobiotic extrusion pumps, was substituted with that of PlcH. The antibiotic susceptibility of oprM-deficient cells expressing the hybrid protein PlcH-OprM was evaluated using the agar dilution method. RESULTS: The PlcH-OprM-expressing cells showed resistance to various MexAB-OprM substrate antibiotics. To evaluate the translocation route of PlcH-OprM, tatC encoding an indispensable component of the Tat machinery was knocked out in oprM-deficient cells. The tatC-oprM double mutant expressing PlcH-OprM exhibited antibiotic hypersusceptibility like the oprM-deficient cells, indicating that PlcH-OprM was translocated across the inner membrane exclusively through the Tat system. CONCLUSIONS: This system can be used for the screening of Tat system inhibitors and will be an excellent model for the study of secretion and biogenesis of the ß-barrel outer membrane proteins.
Assuntos
Adenosina Trifosfatases/metabolismo , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Antibacterianos/metabolismo , Antibacterianos/farmacologia , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Bactérias/genética , Farmacorresistência Bacteriana/efeitos dos fármacos , Proteínas de Membrana Transportadoras/deficiência , Proteínas de Membrana Transportadoras/genética , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/metabolismo , Canais de Translocação SEC , Proteínas SecA , Especificidade por Substrato , Xenobióticos/metabolismo , Xenobióticos/farmacologiaRESUMO
This study aimed to determine the antifungal effect of Ethiopian multiflora honey against Candida species isolated from the oral cavity of AIDS patients. Oral rinses were obtained from 13 AIDS patients and cultured on CHROMagar plates at 37°C for 48 hours. Candida species were identified by microbiological and molecular techniques. The antifungal effect of the honey sample on Candida was investigated by an agar dilution technique. Susceptibility of the Candida species to fluconazole was tested following a semi-modified microdilution method. Growth of both fluconazole-susceptible and -resistant Candida species was inhibited with a minimum fungicidal concentration (MFC) of 35-40% (v/v) honey. The MFC of different Candida species was not significantly different (P > 0.05). From the total of 25 Candida isolates tested for susceptibility, 11 (44%), eight (32%) and six (24%) of the isolates were sensitive (minimum inhibitory concentrations [MICs] < 8 µg/mL), susceptible (dose-dependent: MICs 16-32 µg/mL) and resistant (MICs > 64 µg/mL) to fluconazole, respectively. Ethiopian multiflora honey has antifungal activity against fluconazole-resistant Candida species isolated from the oral cavity of AIDS patients. This supports the existing folkloric practice of using honey to treat oral lesions. Nevertheless, identification of the bioactive agents in honey, their clinical evaluation and pharmacological standardization are crucial.
Assuntos
Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Antifúngicos/farmacologia , Candida/efeitos dos fármacos , Mel , Boca/microbiologia , Candida/isolamento & purificação , Candidíase/microbiologia , Candidíase/terapia , Candidíase/virologia , Relação Dose-Resposta a Droga , Farmacorresistência Fúngica , Etiópia , Fluconazol/farmacologia , Humanos , Testes de Sensibilidade Microbiana , Nefelometria e TurbidimetriaRESUMO
INTRODUCTION: Ceragenin CSA-13 is a bile-acid-based mimic of endogenous antimicrobial peptides and shares a mechanism of action with many of these antimicrobial agents. Because CSA-13 is not peptide based, it is not a substrate for the proteases that are found in the oral cavity, which are capable of degrading antimicrobial peptides. Furthermore, the simplicity of the ceragenins makes them easier to prepare and purify than antimicrobial peptides. In this study, we examined the antimicrobial activities of CSA-13 against oral pathogens and found that this compound was bactericidal against all of the strains tested. METHODS: The strains used were isolates of Streptococcus mutans and Porphyromonas species. Minimum inhibitory concentrations (MIC) were determined using agar dilution methods. In susceptibility testing, viable counts were determined after incubation with CSA-13. RESULTS: CSA-13 was potent against all 23 strains tested with MICs of 1-8 microg/ml for S. mutans and 1-16 microg/ml for 24 strains of the genus Porphyromonas. The MIC(50) was 2 and the MIC(90) was 8 mug/ml for S. mutans. MIC ranges for protease-positive P. gingivalis and P. cangingivalis were 2-16 microg/ml, and 1-2 microg/ml for protease-negative P. circumdentaria. CSA-13 interacted with lipopolysaccharide-sensitized erythrocytes at a concentration of 5.0-20.0 microg/ml. CONCLUSION: CSA-13 displays broad-spectrum activity against cariogenic and periodontopathic bacteria. CSA-13 was effective against protease-positive Porphyromonas. It was shown to bind to erythrocytes coated with lipopolysaccharide and lipoteichoic acid from diverse bacterial strains. These results suggest that CSA-13 may be useful for the prevention and treatment of oral microbial diseases.
Assuntos
Anti-Infecciosos Locais/farmacologia , Porphyromonas/efeitos dos fármacos , Esteroides/farmacologia , Streptococcus mutans/efeitos dos fármacos , Sequência de Aminoácidos , Peptídeos Catiônicos Antimicrobianos/síntese química , Peptídeos Catiônicos Antimicrobianos/farmacologia , Placa Dentária/microbiologia , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Humanos , Dados de Sequência Molecular , Doenças Periodontais/microbiologia , Esteroides/síntese química , Esteroides/químicaRESUMO
Human neuroblastoma remains enigmatic because it often shows spontaneous regression and aggressive growth. The prognosis of advanced stage of sporadic neuroblastomas is still poor. Here, we investigated whether genomic and molecular signatures could categorize new therapeutic risk groups in primary neuroblastomas. We conducted microarray-based comparative genomic hybridization (array-CGH) with a DNA chip carrying 2464 BAC clones to examine genomic aberrations of 236 neuroblastomas and used in-house cDNA microarrays for gene-expression profiling. Array-CGH demonstrated three major genomic groups of chromosomal aberrations: silent (GGS), partial gains and/or losses (GGP) and whole gains and/or losses (GGW), which well corresponded with the patterns of chromosome 17 abnormalities. They were further classified into subgroups with different outcomes. In 112 sporadic neuroblastomas, MYCN amplification was frequent in GGS (22%) and GGP (53%) and caused serious outcomes in patients. Sporadic tumors with a single copy of MYCN showed the 5-year cumulative survival rates of 89% in GGS, 53% in GGP and 85% in GGW. Molecular signatures also segregated patients into the favorable and unfavorable prognosis groups (P=0.001). Both univariate and multivariate analyses revealed that genomic and molecular signatures were mutually independent, powerful prognostic indicators. Thus, combined genomic and molecular signatures may categorize novel risk groups and confer new clues for allowing tailored or even individualized medicine to patients with neuroblastoma.
Assuntos
Perfilação da Expressão Gênica , Neuroblastoma/diagnóstico , Neuroblastoma/genética , Análise de Sequência com Séries de Oligonucleotídeos , Criança , Pré-Escolar , Aberrações Cromossômicas , Cromossomos Humanos Par 17 , Análise por Conglomerados , Amplificação de Genes , Humanos , Lactente , Recém-Nascido , Proteína Proto-Oncogênica N-Myc , Neuroblastoma/classificação , Neuroblastoma/mortalidade , Proteínas Nucleares/genética , Proteínas Oncogênicas/genética , Prognóstico , Risco , Análise de SobrevidaRESUMO
Retinoic acid (RA) has been shown to induce neuronal differentiation and/or apoptosis, and is widely used as a chemotherapeutic agent for treating the patients with neuroblastoma. However, the therapeutic effect of RA is still limited. To unveil the molecular mechanism(s) inducing differentiation and apoptosis in neuroblastoma cells, we compared CHP134 and NB-39-nu cell lines, in which all-trans-RA (ATRA) induces apoptosis, with LA-N-5 and RTBM1 cell lines, in which it induces neuronal differentiation. Here, we found that Bcl-2 was strongly downregulated in CHP134 and NB-39-nu cells, whereas it was abundantly expressed in LA-N-5 and RTBM1 cells. ATRA-mediated apoptosis in CHP134 and NB-39-nu cells was associated with a significant activation of caspase-9 and caspase-3 as well as cytoplasmic release of cytochrome c from mitochondria in a p53-independent manner. Enforced expression of Bcl-2 significantly inhibited ATRA-mediated apoptosis in CHP134 cells. In addition, treatment of RTBM1 cells with a Bcl-2 inhibitor, HA14-1, enhanced apoptotic response induced by ATRA. Of note, two out of 10 sporadic neuroblastomas expressed bcl-2 at undetectable levels and underwent cell death in response to ATRA in primary cultures. Thus, our present results suggest that overexpression of Bcl-2 is one of the key mechanisms to give neuroblastoma cells the resistance against ATRA-mediated apoptosis. This may provide a new therapeutic strategy against the ATRA-resistant and aggressive neuroblastomas by combining treatment with ATRA and a Bcl-2 inhibitor.
Assuntos
Apoptose/efeitos dos fármacos , Neuroblastoma/fisiopatologia , Proteínas Proto-Oncogênicas c-bcl-2/fisiologia , Tretinoína/farmacologia , Apoptose/fisiologia , Sequência de Bases , Western Blotting , Diferenciação Celular , Linhagem Celular Tumoral , Imunofluorescência , Humanos , Dados de Sequência Molecular , Neuroblastoma/patologia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Differential screening of the genes obtained from cDNA libraries of primary neuroblastomas (NBLs) between the favorable and unfavorable subsets has identified a novel gene BCH motif-containing molecule at the carboxyl terminal region 1 (BMCC1). Its 350 kDa protein product possessed a Bcl2-/adenovirus E1B nineteen kDa-interacting protein 2 (BNIP2) and Cdc42GAP homology domain in the COOH-terminus in addition to P-loop and a coiled-coil region near the NH2-terminus. High levels of BMCC1 expression were detected in the human nervous system as well as spinal cord, brain and dorsal root ganglion in mouse embryo. The immunohistochemical study revealed that BMCC1 was positively stained in the cytoplasm of favorable NBL cells but not in unfavorable ones with MYCN amplification. The quantitative real-time reverse transcription-PCR using 98 primary NBLs showed that high expression of BMCC1 was a significant indicator of favorable NBL. In primary culture of newborn mice superior cervical ganglion (SCG) neurons, mBMCC1 expression was downregulated after nerve growth factor (NGF)-induced differentiation, and upregulated during the NGF-depletion-induced apoptosis. Furthermore, the proapoptotic function of BMCC1 was also suggested by increased expression in CHP134 NBL cells undergoing apoptosis after treatment with retinoic acid, and by an enhanced apoptosis after depletion of NGF in the SCG neurons obtained from newborn mice transgenic with BMCC1 in primary culture. Thus, BMCC1 is a new member of prognostic factors for NBL and may play an important role in regulating differentiation, survival and aggressiveness of the tumor cells.
Assuntos
Proteínas de Transporte/genética , Proteínas de Neoplasias/genética , Neuroblastoma/genética , Neuroblastoma/patologia , Sequência de Aminoácidos , Animais , Proteínas de Transporte/fisiologia , Diferenciação Celular , Sobrevivência Celular , Feminino , Perfilação da Expressão Gênica , Biblioteca Gênica , Humanos , Imuno-Histoquímica , Camundongos , Dados de Sequência Molecular , Proteínas de Neoplasias/fisiologia , Prognóstico , Gânglio Cervical Superior/citologiaRESUMO
Differential screening-selected gene aberrative in neuroblastoma (Dan) protein is produced in small neurons of dorsal root ganglia. Thermal and mechanical allodynia and Fos expression in the spinal dorsal horn evoked by inflammation and neuropathic pain were investigated using Dan-deficient mice. Mice showed pain reactions induced by the introduction of complete Freund's adjuvant (CFA) into their hind paw (inflammatory pain model) and after sciatic nerve ligation (neuropathic pain model). In the inflammatory pain model, thermal and mechanical pain thresholds in Dan-deficient mice were significantly higher than those of wild-type mice. The number of Fos-immunoreactive cells in the dorsal horn during the inflammatory period was significantly less in Dan-deficient mice. However, in the neuropathic pain model, no differences in thermal hypersensitivity, mechanical allodynia, or the number of Fos-immunoreactive cells in the dorsal horn were observed between the mice. These data suggest that Dan may be a neuromodulator in inflammatory pain.
Assuntos
Inflamação/metabolismo , Medição da Dor/métodos , Dor/metabolismo , Proteínas/metabolismo , Animais , Proteínas de Ciclo Celular , Citocinas , Temperatura Alta/efeitos adversos , Hiperalgesia/genética , Hiperalgesia/metabolismo , Inflamação/genética , Masculino , Camundongos , Camundongos Knockout , Dor/genética , Proteínas/genéticaRESUMO
Behçet's disease (BD) is a multisystemic inflammatory disorder. Although the cause and pathogenesis of BD are still unclear, there is evidence for genetic, immunologic and infectious factors at the onset or in the course of BD. This review focuses on the functional genomics and immunology of BD. HLA-B51 is the major disease susceptibility gene locus in BD. An increased number of gammadelta T cells in the peripheral blood and in the involved tissues have been reported. However, the T cells at the sites of inflammation appear to be a phenotypically distinct subset. There is also a significant gammadelta T cell proliferative response to mycobacterial 65-kDa heat shock protein peptides. Homologous peptides derived from the human 60-kDa heat shock protein were observed in BD patients. There is evidence that natural killer T cells may also play a role in BD.
Assuntos
Síndrome de Behçet/genética , Síndrome de Behçet/imunologia , Animais , Antígenos de Bactérias/imunologia , Síndrome de Behçet/etnologia , Citocinas/imunologia , Citocinas/metabolismo , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Genoma Humano , Genótipo , Grécia , Antígenos HLA-B/imunologia , Antígenos HLA-B/metabolismo , Antígeno HLA-B51 , Proteínas de Choque Térmico/metabolismo , Humanos , Itália , Japão , Jordânia , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Linfócitos T/imunologia , Linfócitos T/metabolismoRESUMO
This paper reports the effect of the synthesized 27-amino acid sequence in the C-terminal domain of human CAP18 (hCAP18), a human cationic antibacterial protein or cathelicidin, on certain strains belonging to the genera Porophyromonas and Prevotella. The domain binds lipopolysaccharides (LPS) from Porophyromonas gingivalis and Porophyromonas circumdentaria as well as enterobacterial LPS. Two analogues of hCAP18, designated LL/CAP18 and FF/CAP18, were also tested to determine whether additional activity was obtained. The analogue peptides replaced with hydrophobic and cationic amino acid residues showed more potent bactericidal and LPS-binding activities than the original one.
Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Lipopolissacarídeos/antagonistas & inibidores , Família Multigênica/efeitos dos fármacos , Porphyromonas/efeitos dos fármacos , Prevotella/efeitos dos fármacos , Catelicidinas , Humanos , Porphyromonas/classificação , Porphyromonas gingivalis/efeitos dos fármacos , Prevotella intermedia/efeitos dos fármacos , Prevotella melaninogenica/efeitos dos fármacosAssuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Síndrome de Behçet/microbiologia , Lipopolissacarídeos/farmacologia , Streptococcus sanguis/efeitos dos fármacos , Anti-Infecciosos/farmacologia , Catelicidinas , Humanos , Testes de Sensibilidade Microbiana , Streptococcus/efeitos dos fármacosRESUMO
AIMS: To examine the efficacy of liposome oral administration to induce systemic and mucosal immune responses against verotoxin-producing Escherichia coli (VTEC) and the effect of the induced antibodies on the binding of the bacteria to Caco-2 cells. METHODS AND RESULTS: Mice were immunized orally with VTEC antigen and monophosphoryl lipid A (MPL)-containing liposomes composed of dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylserine and cholesterol (1 : 1 : 2, molar ratio) (PS-liposome). After immunization, significant IgA and IgG responses to VTEC were induced in both serum and the intestinal lavage fluid in all mice tested. Furthermore, anti-VTEC IgA and IgG antibodies in the lavage fluid effectively inhibited the adhesion of VTEC to Caco-2 cells. CONCLUSIONS: Oral immunization with liposome-associated E. coli O157:H7 antigen can induce significant systemic and mucosal antibody responses against the bacterial antigen and antibodies produced in the intestinal tract, thus functioning as inhibitors for preventing VTEC infection. SIGNIFICANCE AND IMPACT OF THE STUDY: Oral PS-liposome vaccines containing MPL have the potential usefulness for the induction of a protective mucosal immune response against intestinal diseases.
Assuntos
Anticorpos Antibacterianos/biossíntese , Aderência Bacteriana , Vacinas contra Escherichia coli/imunologia , Escherichia coli/imunologia , Mucosa Intestinal/imunologia , Administração Oral , Animais , Toxinas Bacterianas/biossíntese , Células CACO-2 , Células Cultivadas , Escherichia coli/patogenicidade , Humanos , Imunoglobulina A/biossíntese , Imunoglobulina G/biossíntese , Imunoterapia Ativa , Lipossomos , Camundongos , Camundongos Endogâmicos BALB C , Toxinas Shiga/biossínteseRESUMO
The Dan gene was first identified as the putative rat tumor suppressor gene and encodes a protein structurally related to Cerberus and Gremlin in vertebrates. Xenopus DAN, as with Cerberus and Gremlin, was demonstrated to block bone morphogenetic protein (BMP) signaling by binding BMPs, and to be capable of inducing additional anterior structures by ectopic overexpression in Xenopus embryos. DAN, thus, is suggested to play pivotal roles in early patterning and subsequent organ development, as in the case of other BMP antagonists. In this report, we isolated the chicken counterpart of Dan. Chicken Dan is mainly expressed in the cephalic and somitic mesoderm and several placodes during organ development.
Assuntos
Biossíntese de Proteínas , Proteínas , Proteínas de Xenopus , Sequência de Aminoácidos , Animais , Proteínas de Ciclo Celular , Embrião de Galinha , DNA Complementar/metabolismo , Humanos , Hibridização In Situ , Dados de Sequência Molecular , Homologia de Sequência de Aminoácidos , Fatores de Tempo , Distribuição Tecidual , XenopusRESUMO
The ability of an anti-TNF-alpha antibody to confer protection against enterohaemorrhagic Escherichia coli (EHEC) O157 was investigated in germfree IQI mice. The use of an antibiotic levofloxacin (LVFX) alone or with the antibody was also studied. Protection included an increase in survival rate. Treatment with the anti-TNF-alpha antibody inhibited the histological signs associated with EHEC infection but did not prevent the colonization of EHEC or production of Shiga toxin (Stx). No clinical signs were observed and EHEC was completely eliminated in the mouse model receiving both anti-TNF-alpha antibody and LVFX. Anti-TNF-alpha antibody suppressed inflammatory cytokine response in the mouse kidney and brain by EHEC infection.
Assuntos
Anti-Infecciosos/uso terapêutico , Anticorpos Monoclonais/uso terapêutico , Infecções por Escherichia coli/tratamento farmacológico , Escherichia coli O157/efeitos dos fármacos , Levofloxacino , Ofloxacino/uso terapêutico , Fator de Necrose Tumoral alfa/imunologia , Animais , Anti-Infecciosos/farmacologia , Anticorpos Antibacterianos , Anticorpos Monoclonais/farmacologia , Modelos Animais de Doenças , Infecções por Escherichia coli/microbiologia , Escherichia coli O157/metabolismo , Feminino , Vida Livre de Germes , Imunoglobulina G/farmacologia , Imunoglobulina G/uso terapêutico , Rim/patologia , Masculino , Camundongos , Ofloxacino/farmacologia , Toxina Shiga/biossíntese , Análise de Sobrevida , Fatores de TempoRESUMO
Differential screening-selected gene aberrative in neuroblastoma(DAN) belongs to a novel gene family(DAN family) that includes the head-inducing factor, Cerberus, and dorsaling factor, Gremlin. It has been suggested that DAN family members control diverse processes in growth, development and the cell cycle. Here, we demonstrate that DAN is produced in the small neurons of the dorsal root ganglion(DRG) and transported to the nerve terminals in the spinal dorsal horn in adult rats. Furthermore, intrathecal injection of an antibody to DAN suppressed pain sensations induced by the application of complete Freund's adjuvant and carageenan into the rat hindpaw, and the amount of DAN mRNA in the DRG neurons and of DAN in the spinal dorsal horn were increased in the inflammatory models. These data suggest that DAN in a novel neurotransmitter and/or modulator in the primary sensory nerve fibers for pain sensation.
Assuntos
Dor/fisiopatologia , Proteínas/fisiologia , Animais , Anticorpos/uso terapêutico , Gânglios Espinais/metabolismo , Inflamação/etiologia , Inflamação/fisiopatologia , Proteínas do Tecido Nervoso , Neurônios Aferentes/fisiologia , Neurotransmissores/imunologia , Neurotransmissores/metabolismo , Neurotransmissores/fisiologia , Dor/etiologia , Manejo da Dor , Proteínas/imunologia , Proteínas/metabolismo , RatosRESUMO
BACKGROUND: Neuroblastoma (NBL) is one of the most common solid malignancies in childhood and is derived from the sympathetic precursor cells. Although p53, a tumor suppressor, has been reported to be rarely mutated in NBLs, it is sequestered abnormally in the cytoplasm of the NBL cell. The mechanism and functional role of the abnormal intracellular localization of p53 remain unclear. PROCEDURE: Here, we established an in vitro system of apoptosis model using a NBL cell line CHP134 which also showed a cytoplasmic sequestration of p53. The treatment of the cells with 1 or 5 microM all-trans retinoic acid (RA) induced moderate neurite outgrowth followed by massive death of CHP134 cells by days 5 to 6. RESULTS: TUNEL staining showed that the cell death was due to apoptosis. Immunofluorescent stain demonstrated that p53 was strongly positive in the nucleus on day 5, which was accompanied with induction of p21WAF1. In addition, expression of caspase-3 was also increased during the cell death. Intriguingly, the RA treatment induced expression of Ret tyrosine kinase receptor in CHP134 cells. CONCLUSIONS: The addition of ligands, glial cell line-derived neurotrophic factor (GDNF) and neurturin (NTN), inhibited apoptosis as well as nuclear accumulation of p53 in the cell. The present results suggest that the RA-induced apoptosis of NBL cells is associated with activation of both the caspase cascade and the p53-mediated pathway with its nuclear translocation. The neurotrophic signal through the GDNF-Ret system may prevent the neuronal cell death.
Assuntos
Apoptose/efeitos dos fármacos , Proteínas de Drosophila , Proteínas de Neoplasias/fisiologia , Fatores de Crescimento Neural/farmacologia , Proteínas do Tecido Nervoso/farmacologia , Neuroblastoma/patologia , Proteínas Proto-Oncogênicas/efeitos dos fármacos , Receptores Proteína Tirosina Quinases/efeitos dos fármacos , Proteína Supressora de Tumor p53/fisiologia , Transporte Ativo do Núcleo Celular , Caspase 3 , Caspases/biossíntese , Caspases/genética , Núcleo Celular/metabolismo , Inibidor de Quinase Dependente de Ciclina p21 , Ciclinas/biossíntese , Ciclinas/genética , Indução Enzimática/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Fator Neurotrófico Derivado de Linhagem de Célula Glial , Receptores de Fator Neurotrófico Derivado de Linhagem de Célula Glial , Humanos , Proteínas de Neoplasias/biossíntese , Proteínas de Neoplasias/efeitos dos fármacos , Proteínas de Neoplasias/genética , Proteínas do Tecido Nervoso/fisiologia , Neuritos/efeitos dos fármacos , Neurturina , Proteínas Proto-Oncogênicas/fisiologia , Proteínas Proto-Oncogênicas c-ret , Receptores Proteína Tirosina Quinases/fisiologia , Transdução de Sinais/efeitos dos fármacos , Tretinoína/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/patologiaRESUMO
Structural studies were carried out on an O-antigenic polysaccharide moiety derived from Porphyromonas circumdentaria NCTC 12469, a reference strain of Porphyromonas species. The polysaccharide chain was composed of D-glucose, D-galactose, N-acetyl-D-glucosamine, and N-acetyl-D-galactosamine in a molar ratio of 1:2:1:1. On the basis of results from 1H- and 13C-NMR spectroscopic analyses including COSY, TOCSY, and HMQC experiments together with results of Smith degradation, methylation analysis, and partial acid hydrolysis, it is concluded that the polysaccharide chain has a pentasaccharide repeating unit of -->6)-beta-D-Glcp-(1-->6)-beta-D-Galp-(1-->3)-beta-D-GlcpNAc-(1-->3)-beta-D-GalpNAc-(1-->. The immunoreaction between P. circumdentaria LPS and the corresponding antiserum was strongly inhibited by the pentasaccharide fragment (Glc-Gal-Gal-GlcNAc-GalNAc) isolated from partial acid hydrolysis of the above polysaccharide, suggestive of O-antigen specific antibodies in the used antiserum.
Assuntos
Antígenos O/química , Porphyromonas/química , Porphyromonas/imunologia , Animais , Sequência de Carboidratos , Humanos , Hidrólise , Espectroscopia de Ressonância Magnética , Dados de Sequência Molecular , Porphyromonas/patogenicidadeRESUMO
The purpose of the present study was to (1) determine the standard value of maximum bite force and to (2) compare the maximum bite force of the elderly between healthy and frail subjects. Subjects included 349 healthy elderly individuals (149 males, 200 females) and 24 frail elderly individuals (seven males, 17 females) ranging from 65 to 74 years of age. Maximum bite force was evaluated using a Dental Prescale system. The maximum bite force of the healthy subjects was significantly higher than that of the frail subjects in both males (P=0.020) and females (P=0.015). However, no significant difference was observed in the number of present teeth between the healthy and frail subjects. Median of maximum bite force in healthy males was 408.0 N, and that of the healthy females was 243.5 N. These results suggest that the frail elderly have latent bite force problems.
Assuntos
Força de Mordida , Dentição , Idoso Fragilizado , Idoso , Feminino , Humanos , Indicadores e Reagentes , Arcada Edêntula/fisiopatologia , Arcada Parcialmente Edêntula/fisiopatologia , Masculino , Pressão , Fatores Sexuais , Estatísticas não ParamétricasRESUMO
In order to clarify the presence and verotoxin (VT) inhibitory activity of globotriaosylceramide (Gb3) in bovine milk, we analyzed neutral glycosphingolipids (GSLs) from bovine milk and investigated the inhibitory effect of bovine milk Gb3 on the cytotoxicity of VT2. Five species of neutral GSLs, designated as N-1, N-2, N-3, N-4, and N-5, were separated on thin-layer chromatography (TLC). N-1, N-2, and N-3 showed the same mobility as glucosylceramide, lactosylceramide, and Gb3 on the TLC plate, respectively. N-4 and N-5 GSLs migrated below globoside on the TLC plate. N-3 GSL having the same TLC mobility as Gb3 from bovine milk was immunologically identified as Gb3 by monoclonal antibody against Gb3, anti-CD77 monoclonal antibody. Furthermore, the effect of bovine milk Gb3 on VT2-induced cytotoxicity was investigated. We found that treatment of VT2 with bovine milk Gb3 can reduce the cytotoxic effect of VT2.
Assuntos
Escherichia coli O157/patogenicidade , Leite/química , Toxinas Shiga/antagonistas & inibidores , Triexosilceramidas/farmacologia , Adsorção , Animais , Bovinos , Chlorocebus aethiops , Lipossomos , Toxinas Shiga/toxicidade , Triexosilceramidas/isolamento & purificação , Células VeroRESUMO
We studied the synergistic effects of Japanese green tea extract (JGTE) and levofloxacin (LVFX) against enterohemorrhagic Escherichia coli (EHEC) infection in a gnotobiotic mouse model. Mice fed on JGTE conferred a significant degree of protection against an oral challenge with EHEC. Complete elimination of the bacteria from the mice, was however, difficult. The combination of JGTE and LVFX increased the survival rate and reduced damage to target organs. Thus, dietary supplementation with JGTE improved the therapeutic effects of antibiotic treatment.
Assuntos
Anti-Infecciosos/farmacologia , Catequina/farmacologia , Infecções por Escherichia coli/tratamento farmacológico , Levofloxacino , Ofloxacino/farmacologia , Chá/química , Animais , Contagem de Colônia Microbiana , Sinergismo Farmacológico , Ensaio de Imunoadsorção Enzimática , Escherichia coli O157/efeitos dos fármacos , Escherichia coli O157/metabolismo , Fezes/microbiologia , Feminino , Vida Livre de Germes , Masculino , Camundongos , Camundongos Endogâmicos ICR , Ofloxacino/uso terapêutico , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Toxina Shiga/metabolismoRESUMO
We report a 33-year-old man with bilateral facial paralysis due to neuroborreliosis. About three weeks after rhinorrhea and fever lasting four days, he noticed fatigue in the legs and paresthesia in all four extremities. Another week later, he developed paresthesia in his tongue and bilateral facial muscle weakness, and was admitted to our hospital. On admission, neurological examination revealed moderate bilateral facial muscle weakness, mild paresthesia in the tongue and four extremities, and decreased Achilles tendon reflex bilaterally. Mild pleocytosis and increased protein were found in the cerebrospinal fluid (CSF). IgM antibodies that reacted with the antigens of Borrelia garinii and Borrelia afzelii were found in his serum. Clinically and serologically, he was thus diagnosed as having neuroborreliosis. Brain MRI revealed gadolinium-enhanced lesions of the bilateral facial nerves in the facial nerve canal portion. After three weeks of treatment with 100 mg/day doxycycline and 2 g/day ceftriaxone sodium, his symptoms and CSF abnormalities were rapidly improved. Although facial nerve paralysis is a major symptom of neuroborreliosis, the present report is the first to detect the inflammatory lesions of the facial nerves in the facial nerve canal portion by MRI.