RESUMO
The purpose of this study was to investigate the changes in tongue-palatal contact patterns using electropalatography (EPG) before and after sagittal split ramus osteotomy (SSRO) in patients with mandibular prognathism. Nine clients who underwent SSRO for mandibular setback and seven control subjects were participated in this study. Tongue-palatal contact patterns for /t/, /s/ and /k/ production were investigated using EPG before surgery and 3 months after surgery. The mean value of whole total of palate contact (WT) in the maximum contact frame was examined before and after SSRO. The correlation quantity between the change of center of gravity (COG) value and the amount of mandibular setback was also evaluated. The mean value of WT for /t/ and /s/ significantly increased after SSRO, and the EPG pattern became normal. However, a remarkable change in WT for /k/ was not observed, and the mean value was significantly larger in the SSRO group before and after surgery than in the control group. A negative correlation between COG variation and the amount of mandibular setback for /t/ and positive correlation for /s/ was observed. This study demonstrated that tongue-palatal contact patterns for /t/ and /s/ articulation improved clearly after SSRO. There was a significant correlation between COG variation and the amount of mandibular setback. However, no significant change was detected through perceptual assessment before and after SSRO. Further investigation is needed to determine whether these results will change over time.
Assuntos
Eletrodiagnóstico , Mandíbula/cirurgia , Osteotomia Sagital do Ramo Mandibular , Prognatismo/cirurgia , Língua/fisiopatologia , Adulto , Força de Mordida , Feminino , Humanos , Masculino , Mandíbula/anatomia & histologia , Mandíbula/fisiopatologia , Prognatismo/diagnóstico por imagem , Prognatismo/fisiopatologia , Propriocepção , Fatores de Tempo , Língua/anatomia & histologia , Resultado do Tratamento , Adulto JovemRESUMO
STUDY DESIGN: A case-control investigation. OBJECTIVES: The objective of this study was to quantitatively study impaired ability to appropriately adjust pinch strength while holding a small object in patients with cervical spondylotic myelopathy (CSM). SETTING: Kochi Medical School Hospital, Japan. METHODS: The subjects consisted of 19 CSM patients who had frequent episodes of failing to grasp and hold small objects in their daily life (Group A), 13 CSM patients who did not experience such episodes (Group B) and 16 healthy subjects (Control Group). We continuously measured the dynamic internal pressure of a pneumatic rubber object called a blower pinched by the subject, following two different sets of instructions: (1) pinching with eyes open and with the minimal strength required to prevent dropping; and (2) maintaining a constant pinch strength at given levels with eyes closed. RESULTS: Compared with the other two groups, Group A subjects used a significantly (P<0.01) greater pinch strength to avoid dropping the blower held with eyes open and showed a significantly (P<0.01) greater deviation in pinch strength from the baseline values with eyes closed. These tendencies in Group A showed a significant correlation with the tactile perception threshold of the digits (P<0.01) but not with impairment of rapid repetitive movements of the digits that reflects spasticity. CONCLUSION: Our technique applied to CSM patients helps assess functional integrity primarily, if not exclusively, of the fasciculus cuneatus mediating the feedback signals from proprioceptive and cutaneous receptors in the digits, which are otherwise difficult to evaluate quantitatively.
Assuntos
Retroalimentação Sensorial/fisiologia , Força da Mão/fisiologia , Doenças da Medula Espinal/complicações , Espondilose/complicações , Idoso , Idoso de 80 Anos ou mais , Vértebras Cervicais , Avaliação da Deficiência , Feminino , Mãos/fisiopatologia , Humanos , Japão , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-IdadeRESUMO
The influence of locally different species interactions on trait evolution is a focus of recent evolutionary studies. However, few studies have demonstrated that geographically different pollinator-mediated selection influences geographic variation in floral traits, especially across a narrow geographic range. Here, we hypothesized that floral size variation in the Japanese herb Prunella vulgaris L. (Lamiaceae) is affected by geographically different pollinator sizes reflecting different pollinator assemblages. To evaluate this hypothesis, we posed two questions. (1) Is there a positive correlation between floral length and the proboscis length of pollinators (bumblebees) across altitude in a mountain range? (2) Does the flower-pollinator size match influence female and male plant fitness? We found geographic variation in the assemblage of pollinators of P. vulgaris along an altitudinal gradient, and, as a consequence, the mean pollinator proboscis length also changed altitudinally. The floral corolla length of P. vulgaris also varied along an altitudinal gradient, and this variation strongly correlated with the local pollinator size but did not correlate with altitude itself. Furthermore, we found that the size match between the floral corolla length and bee proboscis length affected female and male plant fitness and the optimal size match (associated with peak fitness) was similar for the female and male fitness. Collectively, these results suggest that pollinator-mediated selection influences spatial variation in the size of P. vulgaris flowers at a fine spatial scale.
Assuntos
Altitude , Abelhas , Flores , Lamiaceae/fisiologia , Polinização , Animais , HumanosRESUMO
In the Citadel area of Hue City, drainage systems that include canals and ponds are considerable sources of fecal contaminants to inundated water during the rainy season because canals and ponds receive untreated wastewater. It is important to investigate the characteristics of hydraulics and water pollution in canals and ponds. At the canals and ponds, water sampling was conducted during dry and wet weather periods in order to evaluate fecal contamination and to investigate changes in water pollution caused by runoff inflow. Inundated water was also collected from streets during heavy rainfall. At the canals and ponds, concentrations of Escherichia coli and total coliform exceeded the Vietnamese regulation values for surface water in 23 and 24 out of 27 samples (85 and 89%), respectively. The water samples were categorized based on the characteristics of water pollution using cluster analysis. In the rainy season, continuous monitoring was conducted at the canals and ponds using water depth and electrical conductivity (EC) sensors to investigate the dynamic relationship between water level and water pollution. It is suggested that in the canals, high EC meant water stagnation and low EC signified river water inflow. Therefore, EC might be a good indicator of water flow change in canals.
Assuntos
Fezes , Poluentes da Água/isolamento & purificação , Poluição da Água/análise , Cidades , Drenagem Sanitária , Condutividade Elétrica , Enterobacteriaceae/isolamento & purificação , Monitoramento Ambiental , Escherichia coli/isolamento & purificação , Fezes/microbiologia , Água Doce/química , Água Doce/microbiologia , Chuva , Estações do Ano , Vietnã , Movimentos da ÁguaRESUMO
Like other fields of medicine, robotics and mechanization might be introduced into endovascular coil embolization of intracranial aneurysms for effective treatment. We have already reported that coil insertion force could be smaller and more stable when the coil delivery wire is driven mechanically at a constant speed. Another background is the difficulty in synchronizing operators' minds and hands when two operators control the microcatheter and the coil respectively. We have therefore developed a mechanical coil insertion system enabling a single operator to insert coils at a fixed speed while controlling the microcatheter. Using our new system, the operator manipulated the microcatheter with both hands and drove the coil using foot switches simultaneously. A delivery wire force sensor previously reported was used concurrently, allowing the operator to detect excessive stress on the wire. In vitro coil embolization was performed using three methods: simple mechanical advance of the coil; simple mechanical advance of the coil with microcatheter control; and driving (forward and backward) of the coil using foot switches in addition to microcatheter control. The system worked without any problems, and did not interfere with any procedures. In experimental coil embolization, delivery wire control using the foot switches as well as microcatheter manipulation helped to achieve successful insertion of coils. This system could offer the possibility of developing safer and more efficient coil embolization. Although we aim at total mechanization and automation of procedures in the future, microcatheter manipulation and synchronized delivery wire control are still indispensable using this system.
Assuntos
Cateterismo Periférico/instrumentação , Catéteres , Embolização Terapêutica/instrumentação , Aneurisma Intracraniano/cirurgia , Trombólise Mecânica/instrumentação , Robótica/instrumentação , Cateterismo Periférico/métodos , Embolização Terapêutica/métodos , Desenho de Equipamento , Análise de Falha de Equipamento , Humanos , Trombólise Mecânica/métodosRESUMO
An application of sequential automated SPE separation equipment coupled to the quadrupole-based ICPMS instrumentation with a dynamic reaction cell such as a screening test system of (90)Sr and Pu isotopes in environmental samples was developed in this work. So far, during the course of a large number of reports as to various specific radioactivities in environmental samples surveyed at radioactive contaminated area around the Fukushima Daiichi Nuclear Power Plants (FDNPP), there is a much smaller number of reports on (90)Sr and Pu isotopes than that of (134)Cs and (137)Cs since the FDNPP accident, and then it would be expected to develop the simple analysis method of these isotopes instead of radiation measurements currently in use. In particular, a screening for (90)Sr in environmental samples has been accomplished using an isotopic ratio measurement mode in comparison with the characterization on the Solid Phase Elution (SPE) separation between strontium and zirconium isotopes around the mass-90 fraction. As a result, for a trial analysis of environmental samples of a muddy snow water and a soil which were collected at Fukushima, it was found that the present developed system makes it applicable for achieving up to the specific activity levels of several hundreds Bq/kg ((90)Sr) and about 1-2Bq/kg (Pu isotopes) as the screening test system.
Assuntos
Acidente Nuclear de Fukushima , Plutônio/análise , Monitoramento de Radiação/métodos , Poluentes Radioativos/análise , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Radioisótopos de Estrôncio/análise , Japão , Doses de Radiação , Monitoramento de Radiação/instrumentação , Espectrometria de Massas por Ionização por Electrospray/instrumentação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/instrumentaçãoRESUMO
Salt/NaCl has been reported to induce necrosis in gastric mucosal cells, however, the mechanisms for gastric injury by salt are not clarified. In this study, we elucidated whether salt is an oxidative stress inducer via mitochondrial injury on rat gastric epithelial cells (RGM-1) in 300, 450, 650 and 1000 mM of NaCl-contained medium. To clarify whether salt-induced reactive oxygen species (ROS) is derived from mitochondria, we also investigated a salt-induced ROS production in manganese superoxide dismutase overexpressing cells (RGM-MnSOD). MnSOD is a specific scavenger for superoxide anion produced from mitochondria. The results showed that cellular injuries in RGM-MnSOD were significantly less severe than that in normal RGM-1. The electron paramagnetic resonance (EPR) studies also provided an evidence that the salt-derived superoxide production in RGM-MnSOD was less than that in normal RGM-1. These results indicated that salt is not merely a necrotizing factor for gastric epithelial cells, but also an oxidative stress inducer.
Assuntos
Células Epiteliais/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Cloreto de Sódio/farmacologia , Estômago/efeitos dos fármacos , Animais , Morte Celular/efeitos dos fármacos , Linhagem Celular , Células Epiteliais/metabolismo , Células Epiteliais/fisiologia , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/metabolismo , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Oxirredução , Estresse Oxidativo/fisiologia , Ratos , Espécies Reativas de Oxigênio/metabolismo , Estômago/fisiologia , Superóxido Dismutase/metabolismo , Superóxidos/metabolismoRESUMO
BACKGROUND: A low ankle-brachial pressure index (an ABPI value <0.90) is considered predictive of cardiovascular disease, and is widely thought to increase morbidity and mortality in the elderly. However, good nutrition is beneficial both for the health and the ability to resist and recover from the disease. OBJECTIVE: The aim of the present study was to evaluate the relationship between the ankle-brachial pressure index and the nutritional status of the elderly in a city of Kochi prefecture, Japan. METHODS: This was a cross-sectional study in which a total of 100 elderly people, both males and females, were screened for ankle-brachial pressure index (ABPI), nutritional status (through the use of the short form of the mini nutritional assessment), activities of daily living, lifestyle, gait speed (10MWT), postural stability (OLST), and functional mobility (TUG). RESULTS: About 67% of the participants were found to have a normal nutritional status, 27% were at risk of malnutrition, and six percent were classified as malnourished. The mean ABPI of the participants was 1.08±0.10, and three participants had an ABPI lower than 0.90. The ABPI was statistically higher in well nourished participants compared with those at risk of malnutrition or the malnourished. The mean ABPI was significantly higher in non-smokers compared with former smokers. The ABPI was found to correlate negatively with gait speed and with TUG score. CONCLUSION: Well-nourished elderly had a higher normal ankle-brachial pressure index as compared with the malnourished elderly. This study provides supportive evidence for the necessity of adequate nutrition for elderly people.
Assuntos
Índice Tornozelo-Braço , Doenças Cardiovasculares/epidemiologia , Desnutrição/epidemiologia , Avaliação Nutricional , Atividades Cotidianas , Idoso , Idoso de 80 Anos ou mais , Pressão Sanguínea , Doenças Cardiovasculares/fisiopatologia , Estudos Transversais , Feminino , Humanos , Japão , Estilo de Vida , Masculino , Desnutrição/fisiopatologia , Estado Nutricional , Prevalência , Análise de Regressão , Fatores de Risco , Inquéritos e QuestionáriosRESUMO
Easy-to-measure surrogate parameters for water quality indicators are needed for real time monitoring as well as for generating data for model calibration and validation. In this study, a novel linear regression model for estimating total nitrogen (TN) based on two surrogate parameters is proposed based on evaluation of pollutant loads flowing into a eutrophic lake. Based on their runoff characteristics during wet weather, electric conductivity (EC) and turbidity were selected as surrogates for particulate nitrogen (PN) and dissolved nitrogen (DN), respectively. Strong linear relationships were established between PN and turbidity and DN and EC, and both models subsequently combined for estimation of TN. This model was evaluated by comparison of estimated and observed TN runoff loads during rainfall events. This analysis showed that turbidity and EC are viable surrogates for PN and DN, respectively, and that the linear regression model for TN concentration was successful in estimating TN runoff loads during rainfall events and also under dry weather conditions.
Assuntos
Condutividade Elétrica , Monitoramento Ambiental/métodos , Lagos/química , Nitrogênio/química , Poluentes Químicos da Água/química , Japão , Modelos Lineares , Modelos Teóricos , Movimentos da ÁguaRESUMO
Nonsteroidal anti-inflammatory drugs (NSAIDs) often cause gastrointestinal complications such as gastric ulcers and erosions. Recent studies on the pathogenesis have revealed that NSAIDs induce lipid peroxidation in gastric epithelial cells by generating superoxide anion in mitochondria, independently with cyclooxygenase-inhibition and the subsequent prostaglandin deficiency. Although not clearly elucidated, the impairment of mitochondrial oxidative phosphorylation, or uncoupling, by NSAIDs is associated with the generation of superoxide anion. Physiologically, superoxide is immediately transformed into hydrogen peroxide and diatomic oxygen with manganese superoxide dismutase (MnSOD). Rebamipide is an antiulcer agent that showed protective effects against NSAID-induced lipid peroxidation in gastrointestinal tracts. We hypothesized that rebamipide may attenuate lipid peroxidation by increasing the expression of MnSOD protein in mitochondria and decreasing the leakage of superoxide anion in NSAID-treated gastric and small intestinal epithelial cells. Firstly, to examine rebamipide increases the expression of MnSOD proteins in mitochondria of gastrointestinal epithelial cells, we underwent Western blotting analysis against anti-MnSOD antibody in gastric RGM1 cells and small intestinal IEC6 cells. Secondly, to examine whether the pretreatment of rebamipide decreases NSAID-induced mitochondrial impairment and lipid peroxidation, we treated these cells with NSAIDs with or without rebamipide pretreatment, and examined with specific fluorescent indicators. Finally, to examine whether pretreatment of rebamipide attenuates NSAID-induced superoxide anion leakage from mitochondria, we examined the mitochondria from indomethacin-treated RGM1 cells with electron spin resonance (ESR) spectroscopy using a specific spin-trapping reagent, CYPMPO. Rebamipide increased the expression of MnSOD protein, and attenuated NSAID-induced mitochondrial impairment and lipid peroxidation in RGM1 and IEC6 cells. The pretreatment of rebamipide significantly decreased the signal intensity of superoxide anion from the mitochondria. We conclude that rebamipide attenuates lipid peroxidation by increasing the expression of MnSOD protein and decreasing superoxide anion leakage from mitochondria in both gastric and small intestinal epithelial cells.
Assuntos
Alanina/análogos & derivados , Antiulcerosos/farmacologia , Antioxidantes/farmacologia , Células Epiteliais/efeitos dos fármacos , Quinolonas/farmacologia , Superóxido Dismutase/biossíntese , Alanina/farmacologia , Animais , Anti-Inflamatórios não Esteroides/efeitos adversos , Linhagem Celular , Células Epiteliais/fisiologia , Intestino Delgado/citologia , Peroxidação de Lipídeos/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/fisiologia , Ratos , Estômago/citologiaRESUMO
The purpose of this study was to clarify the efficacy and safety of docetaxel and cisplatin as second-line treatment for patients with S-1 refractory advanced gastric cancer. Between 1999 and 2006, 32 patients received docetaxel (60 mg/m²) and cisplatin (60 mg/m²) (Dp regimen) on day 1 every 3 weeks. This regimen was repeated at least three times at 3-week intervals until disease progression or unacceptable toxicity was detected. The overall response rate was 21.9%. Seven patients showed partial response, 17 showed stable disease and 8 showed disease progression. The median survival time was 12.3 months after the start of the first-line treatment. The median survival time and time to progression following the DP regimen was 7.8 months and 4.0 months, respectively. The major adverse effects were leukopenia and neutropnea. Non-hematological toxicities were generally mild to moderate and controllable. this study showed satisfactory therapeutic outcomes for patients with gastric cancer refractory to S- 1 chemotherapy.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Neoplasias Gástricas/tratamento farmacológico , Adulto , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Cisplatino/administração & dosagem , Cisplatino/efeitos adversos , Progressão da Doença , Docetaxel , Combinação de Medicamentos , Resistencia a Medicamentos Antineoplásicos , Feminino , Humanos , Leucopenia/induzido quimicamente , Masculino , Pessoa de Meia-Idade , Ácido Oxônico/farmacologia , Prognóstico , Estudos Retrospectivos , Neoplasias Gástricas/sangue , Taxa de Sobrevida , Taxoides/administração & dosagem , Taxoides/efeitos adversos , Tegafur/farmacologia , Conduta ExpectanteRESUMO
Landing with the knee in a valgus position is recognized as a risk factor for anterior cruciate ligament (ACL) injury. Using linear and non-linear regression analyses, the purpose of this study was to examine the correlation between two-dimensional (2D) knee valgus and three-dimensional (3D) knee kinematics measured during a jump landing task. Twenty-eight female collegiate athletes participated. All participants were required to perform a continuous jump test. The average maximum angles of abduction and internal tibial rotation during landing were measured using the Point Cluster Technique. Average peak knee valgus angle was measured using a 2D approach. Linear and non-linear regression analyses between 2D valgus and 3D knee abduction, and between 2D valgus and 3D internal tibial rotation, were performed. The R2 value between 2D valgus and 3D knee abduction was significantly different from zero and had a moderate correlation for all models, whereas the R2 value between 2D valgus and 3D internal tibial rotation was not significantly different from zero. The 2D approach could be used to screen a specific group of individuals for risk of ACL injury; however, using frontal plane 2D analysis of valgus motion to evaluate internal tibial rotation is not advised.
Assuntos
Articulação do Joelho/fisiologia , Locomoção/fisiologia , Modelos Biológicos , Modelos Estatísticos , Movimento/fisiologia , Esportes com Raquete/fisiologia , Amplitude de Movimento Articular/fisiologia , Voleibol/fisiologia , Adulto , Simulação por Computador , Interpretação Estatística de Dados , Feminino , Humanos , RotaçãoRESUMO
Emdogain has been used clinically for periodontal regeneration, although the underlying molecular mechanisms are not clear at present. In this study, we hypothesized that Emdogain stimulated degradation of type I collagen via osteoblasts. We showed that Emdogain enhanced cell-mediated degradation of type I collagen in an MMP-dependent manner. Although MG-63 cells spontaneously produced a zymogen form of MMP-1, treatment with Emdogain significantly induced the generation of the active form of this enzyme. We demonstrated that MMP-3 was produced from MG63 cells in response to Emdogain in a MEK1/2-dependent manner. Concomitantly, blocking of MEK1/2 activation by U0126 significantly inhibited the generation of the active form of MMP-1 without affecting the total production of this collagenase. These results suggest that Emdogain facilitates tissue regeneration through the activation of the collagenase, MMP-1, that degrades matrix proteins in bone tissue microenvironments.
Assuntos
Colágeno Tipo I/metabolismo , Proteínas do Esmalte Dentário/farmacologia , Metaloproteinase 8 da Matriz/metabolismo , Osteoblastos/metabolismo , Doenças Periodontais/tratamento farmacológico , Regeneração Óssea/fisiologia , Linhagem Celular , Matriz Extracelular/enzimologia , Matriz Extracelular/metabolismo , Humanos , MAP Quinase Quinase 1/metabolismo , MAP Quinase Quinase Quinase 2/metabolismo , Metaloproteinase 1 da Matriz/metabolismo , Doenças Periodontais/metabolismoRESUMO
Faecal prevalence of gastrointestinal bacterial pathogens, including Campylobacter, Escherichia coli O157:H7, Salmonella, Shigella, Yersinia, as well as Arcobacter, were examined in 317 faecal specimens from 44 animal species in Belfast Zoological Gardens, during July-September 2006. Thermophilic campylobacters including Campylobacter jejuni, Campylobacter coli and Campylobacter lari, were the most frequently isolated pathogens, where members of this genus were isolated from 11 animal species (11 of 44; 25%). Yersinia spp. were isolated from seven animal species (seven of 44; 15.9%) and included, Yersinia enterocolitica (five of seven isolates; 71.4%) and one isolate each of Yersinia frederiksenii and Yersinia kristensenii. Only one isolate of Salmonella was obtained throughout the entire study, which was an isolate of Salmonella dublin (O 1,9,12: H g, p), originating from tiger faeces after enrichment. None of the animal species found in public contact areas of the zoo were positive for any gastrointestinal bacterial pathogens. Also, water from the lake in the centre of the grounds, was examined for the same bacterial pathogens and was found to contain C. jejuni. This study is the first report on the isolation of a number of important bacterial pathogens from a variety of novel host species, C. jejuni from the red kangaroo (Macropus rufus), C. lari from a maned wolf (Chrysocyon brachyurus), Y. kristensenii from a vicugna (Vicugna vicugna) and Y. enterocolitica from a maned wolf and red panda (Ailurus fulgens). In conclusion, this study demonstrated that the faeces of animals in public contact areas of the zoo were not positive for the bacterial gastrointestinal pathogens examined. This is reassuring for the public health of visitors, particularly children, who enjoy this educational and recreational resource.
Assuntos
Animais de Zoológico/microbiologia , Bactérias/isolamento & purificação , Fezes/microbiologia , Saúde Pública , Animais , Bactérias/patogenicidade , Campylobacter/isolamento & purificação , Campylobacter/patogenicidade , Controle de Doenças Transmissíveis , Escherichia coli O157/isolamento & purificação , Escherichia coli O157/patogenicidade , Feminino , Irlanda/epidemiologia , Masculino , Prevalência , Salmonella/isolamento & purificação , Salmonella/patogenicidade , Shigella/isolamento & purificação , Shigella/patogenicidade , Especificidade da Espécie , Microbiologia da Água , Yersinia/isolamento & purificação , Yersinia/patogenicidade , ZoonosesAssuntos
Complexo Burkholderia cepacia/isolamento & purificação , Fibrose Cística/microbiologia , Pulmão/microbiologia , Pseudomonas aeruginosa/isolamento & purificação , Água do Mar/microbiologia , Microbiologia da Água , Complexo Burkholderia cepacia/metabolismo , Contagem de Colônia Microbiana , Fibrose Cística/complicações , Humanos , Pseudomonas aeruginosa/metabolismoAssuntos
Cryptosporidium/isolamento & purificação , Contaminação de Alimentos/análise , Lactuca/microbiologia , Reação em Cadeia da Polimerase/métodos , Análise de Sequência de DNA , Animais , Cryptosporidium/genética , Humanos , Dados de Sequência Molecular , Filogenia , Sensibilidade e Especificidade , Esgotos/microbiologiaRESUMO
A genus-specific polymerase chain reaction (PCR)-based assay is developed for the detection and identification of clinically relevant Fusobacterium species, including F. nucleatum and F. necrophorum. Two 16S ribosomal DNA (rDNA) primers, FUSO1 (forward primer: 5'-GAG AGA GCT TTG CGT CC-3' [17-mer]) and FUSO 2 (reverse primer: 5'-TGG GCG CTG AGG TTC GAC -3' [18-mer]) are designed to target conserved regions of the 16S rDNA gene for Fusobacterium spp. Subsequent proof-of-principle studies employing this assay detected Fusobacterium spp. in the faeces of eight (10%) out of 80 patients with suspected gastrointestinal infection. This assay may be used for the genus-specific detection of Fusobacterium spp. from clinical specimens and for subsequent species identification.
Assuntos
DNA Ribossômico/análise , Fusobacterium/genética , Sequência de Bases , Feminino , Fusobacterium necrophorum/genética , Fusobacterium nucleatum/genética , Humanos , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S/genética , Ribotipagem/métodos , Análise de Sequência de DNARESUMO
The primer set for 16S rDNA amplified an amplicon of about 1500 bp in length for three strains of Taylorella equigenitalis (NCTC11184(T), Kentucky188 and EQ59). Sequence differences of the 16S rDNA among the six sequences, including three reference sequences, occurred at only a few nucleotide positions and thus, an extremely high sequence similarity of the 16S rDNA was first demonstrated among the six sequences. In addition, the primer set for 16S-23S rDNA internal spacer region (ISR) amplified two amplicons about 1300 bp and 1200 bp in length for the three strains. The ISRs were estimated to be about 920 bp in length for large ISR-A and about 830 bp for small ISR-B. Sequence alignment of the ISR-A and ISR-B demonstrated about 10 base differences between NCTC11184(T) and EQ59 and between Kentucky188 and EQ59. However, only minor sequence differences were demonstrated between the ISR-A and ISR-B from NCTC11184(T) and Kentucky188, respectively. A typical order of the intercistronic tRNAs with the 29 nucleotide spacer of 5'-16S rDNA-tRNA(Ile)-tRNA(Ala)-23S rDNA-3' was demonstrated in the all ISRs. The ISRs may be useful for the discrimination amongst isolates of T. equigenitalis if sequencing is employed.