Molecular model of shikimate kinase from Mycobacterium tuberculosis

Tuberculosis (TB) resurged in the late 1980s and now kills approximately 3 million people a year. The reemergence of tuberculosis as a public health threat has created a need to develop new anti-mycobacterial agents. The shikimate pathway is an attractive target for herbicides and anti-microbial agents development because it is essential in algae, higher plants, bacteria, and fungi, but absent from mammals. Homologs to enzymes in the shikimate pathway have been identified in the genome sequence of Mycobacterium tuberculosis. Among them, the shikimate kinase I encoding gene (aroK) was proposed to be present by sequence homology. Accordingly, to pave the way for structural and functional efforts towards anti-mycobacterial agents development, here we describe the molecular modeling of M. tuberculosis shikimate kinase that should provide a structural framework on which the design of specific inhibitors may be based. © 2002 Elsevier Science (USA). All rights reserved.

One-step purification of 5-enolpyruvylshikimate-3-phosphate synthase enzyme from Mycobacterium tuberculosis

Currently, there are 8 million new cases and 2 million deaths annually from tuberculosis, and it is expected that a total of 225 million new cases and 79 million deaths will occur between 1998 and 2030. The reemergence of tuberculosis as a public health threat, the high susceptibility of HIV-infected persons, and the proliferation of multi-drug-resistant strains have created a need to develop new antimycobacterial agents. The existence of homologues to the shikimate pathway enzymes has been predicted by the determination of the genome sequence of Mycobacterium tuberculosis. We have previously reported the cloning and overexpression of M. tuberculosis aro A-encoded EPSP synthase in both soluble and active forms, without IPTG induction. Here, we describe the purification of M. tuberculosis EPSP synthase (mtEPSPS) expressed in Escherichia coli BL21(DE3) host cells. Purification of mtEPSPS was achieved by a one-step purification protocol using an anion exchange column. The activity of the homogeneous enzyme was measured by a coupled assay using purified shikimate kinase and purine nucleoside phosphorylase proteins. A total of 53 mg of homogeneous enzyme could be obtained from 1 L of LB cell culture, with a specific activity value of approximately 18 U mg-1. The results presented here provide protein in quantities necessary for structural and kinetic studies, which are currently underway in our laboratory. © 2002 Elsevier Science (USA). All rights reserved.

Effect of an alcoholic diet on dental caries and on Streptococcus of the mutans group: Study in rats

The objective of this study was to evaluate the effects of an alcohol diet on Streptococcus of the mutans group and on dental caries in the oral cavity of rats. Forty animals were divided into 3 groups according to the following liquid diets: 20% ethanol solution (Alcohol Group, AG), 27% sucrose solution (Isocaloric Group, IG), and water (Control Group, CG). After 56 days, samples were collected and plated on Mitis Salivarius Bacitracin agar to assess the number of colony forming units (CFU/mL) of Streptococcus of the mutans group. The animals were sacrificed and the jaws were removed in order to assess the occurrence of dental caries on the smooth and occlusal surfaces using stereomicroscopy. The data were submitted to ANOVA and Tukey test. The average numbers of CFU/mL (10 3) were: 8.17 (AG), 9.78 (IG), and 5.63 (CG). There was no significant difference among the groups for the occurrence of occlusal caries. Regarding smooth surface caries, in the upper jaw, the caries number in the IG (1.58) was similar to that in the AG (2.06) and in the CG (1.14), and the number of caries in the AG was higher than in the CG; in the lower jaw there was significant difference among the 3 groups: AG (1.14), IG (2.00) and CG (0.43). The diets with the alcohol and sucrose solutions presented a tendency of increasing the colonization by Streptococcus of the mutans group and of increasing the occurrence of smooth surface dental caries in rat molars when compared to the control diet.

Respiratory immunohistochemical study in rats exposed to cigarette smoke and alcohol

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of estrogen deficiency combined with chronic alcohol consumption on rat mandibular condyle

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of chronic alcoholism and oestrogen deficiency on the variation of stoichiometry of hydroxyapatite within alveolar bone crest of rats

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo comparativo experimental da injeção perineural do álcool benzílico 0,75% e do álcool etílico absoluto nos nervos palmares de eqüinos

The clinical and histopathological effects of two alcoholic neurolytics were studied in horses. Normal horses were shod with a designed shoe adaptted with 5 screws to produce solar pain. After gait and lameness score analysis, the palmar nerve of 5 horses was injected with 5 ml of 0,75% benzyl alcohol (Group A) and 5 horses were injected with 5 ml of absolute ethyl alcohol (Group B). The animals were submitted to regular lameness evaluation and solar sensibility tests during next six months. The solar sensitivity returned 5 months latter in the group injected with benzyl alcohol 0,75%, while in the group injected with absolute ethyl alcohol, the sole was still desensitized 6 months latter. The histopathological findings showed that the nerve injected with benzyl alcohol 0,75%, resulted in axonotmesis, characterized by axonal nerve degeneration, with possibilities for the nerve conduction recovery. The perineural injection of ethyl alcohol absolute, resulted in neurotmesis with difficult nerve regeneration. It was concluded that chemical neurolysis with alcohol is an option for temporary or permanent nerve blocks in horses.

Efeito da ingestão crônica de álcool na progressão de periodontite induzida em ratos da linhagem Fischer-344

Objective: Understand the effect of chronic alcohol on the progression of periodontitis induced in Fischer-344 rats. Methods: For the study, 22 Fischer-344 rats, two months old were used, divided into groups: alcohol (n=8), ligature (n=7) and control (n=7). On the first day, the animals in the alcohol group were exposed to ingestion of a water solution containing 20% alcohol (size/size), up to day 90. After thirty days from the beginning of the experiment, the animals in the alcohol group and the ligature group were submitted to the placement of a silk thread around the right maxillary second molar. Nothing was performed on the left side, serving as control. All the groups were submitted to euthanasia 60 days after ligature placement. To assess the destruction of periodontitis, a radiographic exam was used to measure the destruction of bone height. Results: The results of the study showed that on the side in which periodontitis was induced, the group that ingested alcohol suffered an increase in destruction, with statistical differences when compared with the ligature and control groups and increased bone destruction in the ligature group when compared to control. Conclusion: Within the limitations of the study, it was concluded that chronic alcohol consumption by Fischer-344 rats led to greater progression of induced periodontitis.

Surface agents' influence on the flexural strength of bilaminated ceramics

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Dessensibilização dos nervos palmares em eqüinos: estudo comparativo experimental do álcool benzílico 0,75% e álcool etílico absoluto (95,5% PA)

Pós-graduação em Medicina Veterinária - FMVZ

Efeito do álcool em osteoblastos de recém-nascidos de ratas submetidas ao consumo crônico de etanol

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência ao consumo crônico de álcool no fêmur de ratos machos e fêmeas

Considering that alcohol has been identified, in the literature, as an evident risk factor for osteoporosis development, inducing loss of bone mass, the aim of this study was to evaluate the effect of the alcohol on the femur of male and female rats. For this purpose, 35 male and 35 female rats, divided into seven groups of five animals per genera were included. The groups were: control group that received food and water as much as they wished; alcohol group in the concentrations of 10, 20 and 30% and the isocalorie: groups corresponding to the concentration of alcohol. After 8 weeks, the animals were sacrificed and evaluated regarding to the body weight and nutritional conditions. Unilateral femurs were removed and analyzed in relation to the length, width, and thickness of the bone cortical, trabeculae percentage and optical density. Data obtained were analyzed by ANOVA (Tukey; p<0.05). Regarding to the weight, loss of weight was observed in the animals of the group alcohol 20% among females and 30% among the males. In relation to the nutritional conditions, the females with the dosages of 10, 20 and 30% of alcohol swallowed a mean of 25, 39 and 53% of Kcall and the males 22, 36 and 45 Kcal Jday coming from the alcohol. Alcohol reduced the length of the femur only in the group MG6. Mesio-distal width was higher in the group FG6 although the antero-posterior width was not modified. The cortical thickness among males and females was not altered however the percentage of trabeculae bone was reduced among females alcohol 30% and among males with the three alcoholic concentrations. Optical density was reduced in the concentration of 30% among the males and females. Under the experimental conditions, it could be concluded that the effect of alcohol was more evident among the males and in the trabeculae bone, and that the alcoholic concentration of 30% was the most harmful for the bone tissue, ....

O efeito do consumo abusivo de alvool no torque de remoção de implantes de titânio

Studies have reported that alcohol may lead to imbalance in bone formation and resorption, however, its effects on osseointegration of titanium implants continues to be an inconclusive subject. In this context, the aim of this study was to make a biomechanical evaluation of the effect of abusive alcohol consumption on the removal torque of osseointegrated titanium implants. Male Wistar rats (n=30) were divided into two experimental groups (15 each) receiving only water (Control) or 36% alcohol solution oral administration. Thirty days later, all animals were submitted to titanium implant (2.2 mm x 4 mm) placement in the right and left tibiae. The surgical alveoli were prepared with a 2 mm drill mounted in a counter-angle hand-piece (20:1 ratio, 35 Ncm torque at 1200 rpm) under abundant cooling. Five animals from each group were euthanized at 15, 30, and 60 days. Tibiae were submitted to reverse torque analysis. Data obtained were submitted to statistical analysis by the non-parametric Kruskal-Wallis and Dunn Tests (p < 0.05). Animals in the alcohol group presented lower removal torque values when compared with control group animals for all periods tested (p < 0.05). It can be concluded that abusive alcohol consumption can reduce the removal torque of titanium implants placed in rat tibiae, suggesting that alcohol may interfere in the osseointegration process of titanium implants.

O efeito do consumo abusivo de álcool no torque de remoção de implante de titânio

Studies have reported that alcohol may lead to imbalance in bone formation and resorption, however, its effects on osseointegration of titanium implants continues to be an inconclusive subject. In this context, the aim of this study was to make a biomechanical evaluation of the effect of abusive alcohol consumption on the removal torque of osseointegrated titanium implants. Male Wistar rats (n=30) were divided into two experimental groups (15 each) receiving only water (Control) or 36% alcohol solution oral administration. Thirty days later, all animals were submitted to titanium implant (2.2 mm x 4 mm) placement in the right and left tibiae. The surgical alveoli were prepared with a 2 mm drill mounted in a counter-angle hand-piece (20:1 ratio, 35 Ncm torque at 1200 rpm) under abundant cooling. Five animals from each group were euthanized at 15, 30, and 60 days. Tibiae were submitted to reverse torque analysis. Data obtained were submitted to statistical analysis by the non-parametric Kruskal-Wallis and Dunn Tests (p < 0.05). Animals in the alcohol group presented lower removal torque values when compared with control group animals for all periods tested (p < 0.05). It can be concluded that abusive alcohol consumption can reduce the removal torque of titanium implants placed in rat tibiae, suggesting that alcohol may interfere in the osseointegration process of titanium implants.

Alcohol screening instruments in elderly male: a population-based survey in metropolitan São Paulo, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)