RESUMO
BACKGROUND: Most older drivers continue to drive as they age. To maintain safe and independent transport, mobility is important for all individuals, but especially for older drivers. OBJECTIVE: The objective of this study was to investigate whether automatic transmission, compared with manual transmission, may improve the driving behavior of older drivers. METHOD: In total, 31 older drivers (mean age 75.2 years) and 32 younger drivers - used as a control group (mean age 39.2 years) - were assessed twice on the same fixed route; once in a car with manual transmission and once in a car with automatic transmission. The cars were otherwise identical. The driving behavior was assessed with the Ryd On-Road Assessment driving protocol. Time to completion of left turns (right-hand side driving) and the impact of a distraction task were measured. RESULTS: The older group had more driving errors than the younger group, in both the manual and the automatic transmission car. However, and contrary to the younger drivers, automatic transmission improved the older participants' driving behavior as demonstrated by safer speed adjustment in urban areas, greater maneuvering skills, safer lane position and driving in accordance with the speed regulations. CONCLUSION: Switching to automatic transmission may be recommended for older drivers as a means to maintain safe driving and thereby the quality of their transport mobility.
Assuntos
Envelhecimento/psicologia , Condução de Veículo/psicologia , Automóveis , Adulto , Idoso , Idoso de 80 Anos ou mais , Humanos , Pessoa de Meia-Idade , Destreza Motora , Análise e Desempenho de TarefasRESUMO
The aim of this study was to recombinantly produce and purify Helicobacter pylori adhesin A (HpaA) from Escherichia coli and compare it to purified native H. pylori HpaA, for potential use as a vaccine antigen. The hpaA gene was cloned from H. pylori, transferred to two different expression vectors, and transformed into E. coli. Expression of rHpaA was analysed by immunoblot, inhibition ELISA, and semi-quantitative dot-blot. Using affinity chromatography, rHpaA was purified from E. coli and native HpaA from H. pylori. The binding of both purified proteins to sialic acid was analysed and antibody titres to native and rHpaA were compared after intraperitoneal immunisation of C57/Bl mice. The rHpaA protein was highly expressed in E. coli from both vectors. Purified recombinant and native HpaA bound similarly to fetuin but also to the non-sialylated asialofetuin. Both native HpaA and rHpaA induced comparable amounts of specific antibodies in serum after immunisation and they were identical in double immunodiffusion. In conclusion, rHpaA was successfully produced in E. coli. Purified rHpaA showed biological properties similar to those of native HpaA isolated from H. pylori and may therefore be further used as an antigen in the development of a vaccine against H. pylori infection.
Assuntos
Adesinas Bacterianas/imunologia , Adesinas Bacterianas/metabolismo , Helicobacter pylori/metabolismo , Adesinas Bacterianas/genética , Adesinas Bacterianas/farmacologia , Animais , Anticorpos Antibacterianos/biossíntese , Anticorpos Antibacterianos/sangue , Anticorpos Monoclonais/imunologia , Western Blotting , Cromatografia de Afinidade , Clonagem Molecular , DNA Bacteriano/química , DNA Bacteriano/genética , Ensaio de Imunoadsorção Enzimática , Escherichia coli/química , Escherichia coli/genética , Escherichia coli/metabolismo , Feminino , Helicobacter pylori/genética , Helicobacter pylori/imunologia , Imunização , Imunodifusão , Camundongos , Camundongos Endogâmicos C57BL , Ácido N-Acetilneuramínico/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , alfa-Fetoproteínas/imunologia , alfa-Fetoproteínas/metabolismoRESUMO
The expression of the virulence-associated genes ureA, encoding the urease subunit A, and nap, encoding the neutrophil activating protein, in Helicobacter pylori grown both in the stomach of C57/Bl6 mice and in Brucella broth was quantified by quantitative competitive reverse transcriptase-PCR using a homologous RNA standard (competitor) and an external standard (16S rRNA). The results showed that the ureA and nap transcripts were increased up to 15 and 80 times, respectively, in vivo compared to in vitro. The transcription of ureA and nap also differed in that ureA showed highest expression early in infection in mice whereas nap transcription was variable throughout the 18-week infection period.
Assuntos
Proteínas de Bactérias/genética , Expressão Gênica , Helicobacter pylori/genética , Urease/genética , Animais , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Haemophilus ducreyi and Klebsiella (Calymmatobacterium) granulomatis are sexually transmitted bacteria that cause characteristic, persisting ulceration on external genitals called chancroid and granuloma inguinale, respectively. Those ulcers are endemic in developing countries or exist, as does granuloma inguinale, only in some geographic "hot spots."H. ducreyi is placed in the genus Haemophilus (family Pasteurellacae); however, this phylogenetic position is not obvious. The multiple ways in which the bacterium may be adapted to its econiche through specialized nutrient acquisitions; defenses against the immune system; and virulence factors that increase attachment, fitness, and persistence within genital tissue are discussed below. The analysis of K. granulomatis phylogeny demonstrated a high degree of identity with other Klebsiella species, and the name K. granulomatis comb. nov. was proposed. Because of the difficulty in growing this bacterium on artificial media, its characteristics have not been sufficiently defined. More studies are needed to understand bacterial genetics related to the pathogenesis and evolution of K. granulomatis.
Assuntos
Evolução Molecular , Haemophilus ducreyi/genética , Klebsiella/genética , Doenças Bacterianas Sexualmente Transmissíveis/microbiologia , Animais , Cancroide/genética , Cancroide/microbiologia , Cancroide/transmissão , Variação Genética , Haemophilus ducreyi/patogenicidade , Haemophilus ducreyi/fisiologia , Humanos , Klebsiella/patogenicidade , Klebsiella/fisiologia , Infecções por Klebsiella/genética , Infecções por Klebsiella/microbiologia , Infecções por Klebsiella/transmissão , Filogenia , Fatores de Virulência/genética , Fatores de Virulência/fisiologiaRESUMO
The presence and persistence of enterotoxigenic Escherichia coli (ETEC) is poorly investigated in marine habitats. Here we compared ETEC with the more studied fecal contaminant, Salmonella enterica serotype Typhimurium (S. enterica) and the marine bacteria Vibrio parahaemolyticus. All three species of bacteria were culturable on agar plates during 8 weeks of incubation in seawater. However, the culturability of ETEC was positively affected by low temperature whereas V. parahaemolyticus was negatively affected. High-nutrient conditions favored the growth of ETEC but not the other bacteria. When the bacteria were fed to blue mussels, V. parahaemolyticus inhibited the filtration activity and the ingestion was lower than that of the enterobacteria. On the other hand, the mussels were less efficient in eliminating V. parahaemolyticus and an in vitro study showed that the hemocytes of three different species of bivalves were not able to kill this strain of V. parahaemolyticus. The bactericidal capacity of bivalves was seemingly an efficient elimination pathway for S. enterica and ETEC. This study showed that ETEC in endemic areas should, to the same degree as S. enterica and V. parahaemolyticus, be taken in consideration when assessing the role of marine environments as a source of enteric infection.
Assuntos
Escherichia coli Enterotoxigênica/crescimento & desenvolvimento , Salmonella enterica/crescimento & desenvolvimento , Água do Mar/microbiologia , Vibrio parahaemolyticus/crescimento & desenvolvimento , Animais , Bivalves/imunologia , Bivalves/microbiologia , Hemócitos/imunologia , Temperatura , Raios Ultravioleta , Microbiologia da ÁguaRESUMO
To express high quantities of colonization factor antigen I (CFA/I) derived from enterotoxigenic Escherichia coli (ETEC) for use in ETEC vaccines, the entire CFA/I operon consisting of four genes (cfa-A, -B, -C, -E) was cloned into plasmid expression vectors that could be maintained either with or without antibiotic selection. Expression from the powerful tac promoter was under the control of the lacIq repressor present on the plasmids. Fimbriae were expressed on the surface of both a non-toxigenic E. coli K12 strain and a non-toxigenic strain of Vibrio cholerae following induction with isopropyl-beta-D-thiogalactopyranoside (IPTG). It was found that the recombinant E. coli strains expressed up to 16-fold higher levels of CFA/I fimbriae compared to a reference strain which had previously been shown to be among the highest natural producers of the CFA/I fimbriae among tested wild type ETEC strains. Oral immunization with formalin-killed recombinant E. coli bacteria over-expressing CFA/I induced significantly higher serum IgA and IgG+M antibodies responses compared to the reference strain. Oral immunization with formalin-killed recombinant V. cholerae bacteria also induce strong CFA/I-specific serum IgA and IgG+M responses. We conclude that our constructs may be useful as candidate strains in an oral killed CF-ETEC vaccine.
Assuntos
Anticorpos Antibacterianos/sangue , Escherichia coli Enterotoxigênica/imunologia , Infecções por Escherichia coli/imunologia , Escherichia coli/metabolismo , Proteínas de Fímbrias/genética , Fímbrias Bacterianas/genética , Vibrio cholerae/metabolismo , Administração Oral , Animais , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/imunologia , Escherichia coli/imunologia , Infecções por Escherichia coli/sangue , Feminino , Proteínas de Fímbrias/biossíntese , Formaldeído , Imunização , Esquemas de Imunização , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Camundongos , Camundongos Endogâmicos BALB C , Óperon , Plasmídeos , Engenharia de Proteínas , Proteínas Recombinantes/biossíntese , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/imunologia , Vibrio cholerae/imunologiaRESUMO
OBJECTIVE: Peroxynitrite formation, as reflected by nitrotyrosine expression, is low in Helicobacter pylori-infected Mongolian gerbils despite pronounced expression of radical-forming enzymes. The aim of the present study was to investigate in vivo whether H. pylori inhibits either one or both of the nitro- and oxyradical formation pathways. MATERIAL AND METHODS: Male Mongolian gerbils were infected with two different H. pylori strains, TN2GF4 and SS1. Six months after inoculation, direct measurement of NO and H2O2 was performed in vivo using electrochemical microsensors positioned in close proximity to the gastric mucosa. RESULTS: In the TN2GF4-infected animals the level of NO was significantly lower than that in controls. No significant difference in NO levels was detected between the SS1-infected group and the controls. H2O2 was significantly increased in the SS1 animals compared with that in controls after 6 months. The H2O2 level in the TN2GF4 group did not differ from that in controls. CONCLUSIONS: The results indicate that H. pylori infection is associated with strain-dependent functional inhibition of both the NO and oxyradical formation pathways in the gastric mucosa.
Assuntos
Gerbillinae/metabolismo , Infecções por Helicobacter/metabolismo , Helicobacter pylori , Peróxido de Hidrogênio/metabolismo , Óxido Nítrico/metabolismo , Animais , Modelos Animais de Doenças , Eletroquímica/métodos , Helicobacter pylori/crescimento & desenvolvimento , Inflamação/fisiopatologia , Masculino , Modelos Biológicos , Antro Pilórico/microbiologia , Gastropatias/microbiologiaRESUMO
OBJECTIVE: For obscure reasons Helicobacter pylori infection of the gastric mucosa is maintained despite a pronounced host defence response. The present study elucidates possible H. pylori-related interference in the oxy- and nitro-radical formation pathways. MATERIAL AND METHODS: Male Mongolian gerbils were infected with two different H. pylori strains, TN2GF4 and SS1. At 3, 6, 12 or 18 months after inoculation, gastric expressions of myeloperoxidase (MPO), inducible nitric oxide synthase (iNOS) and nitrotyrosine were assessed by Western blotting. RESULTS: Expression of both iNOS and MPO was markedly up-regulated in the H. pylori-infected animals compared with non-infected controls. The TN2GF4-infected animals initially (at 3 and 6 months) demonstrated pronounced expression of both iNOS and MPO. The SSI-infected animals exhibited a slower onset with significantly increased iNOS after 12 and 18 months. Nitrotyrosine expression was slightly elevated in the infected groups at 3 and 6 months compared with that in the controls. Nitrotyrosine levels then decreased and were no longer significantly different from those of controls (TN2GF4-infected animals) or were lower (SS1-infected animals) than in the controls. CONCLUSIONS: The results indicate that peroxynitrite formation as reflected by nitrotyrosine expression is low or even inhibited in chronic H. pylori infection despite pronounced expression of enzymes representing both the oxy- and nitro-radical formation pathways. The results support the theory that H. pylori survival is related to functional inhibition of mucosal enzymatic NO and/or oxy-radical formation.
Assuntos
Mucosa Gástrica/metabolismo , Infecções por Helicobacter/metabolismo , Helicobacter pylori/patogenicidade , Óxido Nítrico Sintase Tipo II/biossíntese , Peroxidase/biossíntese , Tirosina/análogos & derivados , Animais , Western Blotting , Modelos Animais de Doenças , Mucosa Gástrica/microbiologia , Gerbillinae , Infecções por Helicobacter/microbiologia , Masculino , Antro Pilórico/metabolismo , Piloro/metabolismo , Ratos , Índice de Gravidade de Doença , Tirosina/biossínteseRESUMO
Enterotoxigenic Escherichia coli (ETEC) is an important cause of diarrhea among children in developing countries and in travelers to areas of ETEC endemicity. ETEC strains isolated from humans may produce a heat-labile enterotoxin (LT) and two types of the heat-stable enterotoxin STa, called STh and STp, encoded by the estA gene. Two commonly used assay methods for the detection of STa, the infant mouse assay or different enzyme-linked immunosorbent assays, are unable to distinguish between the two subtypes of ST. Different genotypic methods, such as DNA probes or PCR assays, may, however, allow such discrimination. Using gene probes, it has recently been reported that ETEC strains producing STp as the only enterotoxin are not associated with diarrhea. In this study, we have used highly specific PCR methods, including newly designed primers for STh together with previously described STp primers, to compare the relative distribution of STh and STp in ETEC isolated from children with diarrhea in three different geographically distinct areas, i.e., Bangladesh, Egypt, and Guatemala, and from travelers to Mexico and Guatemala. It was found that ETEC strains producing STp were as commonly isolated from cases of diarrhea as strains producing STh both in Egypt and Guatemala, whereas STp strains were considerably less common in Bangladesh. No difference was found in the relative distribution of STh and STp in ETEC strains isolated from travelers with diarrhea and from asymptomatic carriers. Irrespective of ST genotype, the disease symptoms were also similar in both children and travelers.
Assuntos
Toxinas Bacterianas/genética , Diarreia/microbiologia , Enterotoxinas/genética , Escherichia coli/classificação , Viagem , Adulto , Criança , Escherichia coli/patogenicidade , Proteínas de Escherichia coli , Genótipo , HumanosRESUMO
Infection with the human gastric pathogen Helicobacter pylori can give rise to chronic gastritis, peptic ulcer, and gastric cancer. All H. pylori strains express the surface-localized protein HpaA, a promising candidate for a vaccine against H. pylori infection. To study the physiological importance of HpaA, a mutation of the hpaA gene was introduced into a mouse-adapted H. pylori strain. To justify that the interruption of the hpaA gene did not cause any polar effects of downstream genes or was associated with a second site mutation, the protein expression patterns of the mutant and wild-type strains were characterized by two different proteomic approaches. Two-dimensional differential in-gel electrophoresis analysis of whole-cell extracts and subcellular fractionation combined with nano-liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry for outer membrane protein profiling revealed only minor differences in the protein profile between the mutant and the wild-type strains. Therefore, the mutant strain was tested for its colonizing ability in a well-established mouse model. While inoculation with the wild-type strain resulted in heavily H. pylori-infected mice, the HpaA mutant strain was not able to establish colonization. Thus, by combining proteomic analysis and in vivo studies, we conclude that HpaA is essential for the colonization of H. pylori in mice.
Assuntos
Adesinas Bacterianas/metabolismo , Mucosa Gástrica/microbiologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/crescimento & desenvolvimento , Helicobacter pylori/patogenicidade , Adesinas Bacterianas/genética , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Feminino , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Infecções por Helicobacter/imunologia , Helicobacter pylori/genética , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Mutação , ProteomaRESUMO
OBJECTIVE: The Mongolian gerbil is considered as the model of choice when studying adenocarcinoma related to Helicobacter pylori infection. The purpose of this study was to compare two different H. pylori strains and elucidate whether adenocarcinomas developed in gerbils. MATERIAL AND METHODS: Male gerbils were separated into three groups: one control and two groups infected with two different strains of H. pylori, TN2GF4 and SS1. At 3, 6, 12 or 18 months after inoculation 5 animals from each group were sacrificed. The stomach was used for culture, and for histology. RESULTS: Inflammation was seen after 3 months in all the infected animals. In the controls no pathology was found at any time. Intestinal metaplasia was found in both the infected groups. Glands buried in the submucusal layer, changes that might be misinterpreted as adenocarcinoma, were found in 10% of the SS1 and in 65% of the TN2GF4 animals. Adenocarcinoma was not found in any of the gerbils. CONCLUSIONS: All studies claiming to have found H. pylori-induced adenocarcinomas in gerbils describe atypical glands penetrating into the muscularis propria and interpret these as invasive growths due to cancer. An alternative interpretation is that the deranged glandular structures grow in and below the submucosa. It is suggested that atypical glands in the muscularis layer are not enough as a diagnostic criterion for gastric adenocarcinoma. It is concluded that adenocarcinoma has not yet been shown convincingly to develop in Mongolian gerbils infected with H. pylori. Nevertheless, it is a model well suited for studying gastritis, gastric ulcer and premalignant changes such as metaplasia.