Shikimate kinase: A potential target for development of novel antitubercular agents

Tuberculosis (TB) remains the leading cause of mortality due to a bacterial pathogen, Mycobacterium tuberculosis. However, no new classes of drugs for TB have been developed in the past 30 years. Therefore there is an urgent need to develop faster acting and effective new antitubercular agents, preferably belonging to new structural classes, to better combat TB, including MDR-TB, to shorten the duration of current treatment to improve patient compliance, and to provide effective treatment of latent tuberculosis infection. The enzymes in the shikimate pathway are potential targets for development of a new generation of antitubercular drugs. The shikimate pathway has been shown by disruption of aroK gene to be essential for the Mycobacterium tuberculosis. The shikimate kinase (SK) catalyses the phosphorylation of the 3-hydroxyl group of shikimic acid (shikimate) using ATP as a co-substrate. SK belongs to family of nucleoside monophosphate (NMP) kinases. The enzyme is an alpha/beta protein consisting of a central sheet of five parallel beta-strands flanked by alpha-helices. The shikimate kinases are composed of three domains: Core domain, Lid domain and Shikimate-binding domain. The Lid and Shikimate-binding domains are responsible for large conformational changes during catalysis. More recently, the precise interactions between SK and substrate have been elucidated, showing the binding of shikimate with three charged residues conserved among the SK sequences. The elucidation of interactions between MtSK and their substrates is crucial for the development of a new generation of drugs against tuberculosis through rational drug design.

Vertical marginal discrepancy of ceramic copings with different ceramic materials finish lines and luting agents: An in vitro evaluation

Statement of problem. Prosthetic restorations that fit poorly may affect periodontal health and occlusion. Studies that have evaluated the accuracy of fit of ceramic restorations before and after cementation assessed primarily intracoronal restorations.Purpose. This in vitro study evaluated the effect of different finish lines, ceramic manufacturing techniques, and luting agents on the vertical discrepancy of ceramic copings.Material and methods. Two stainless steel molars were prepared for complete crowns with 2 different finish lines (heavy chamfer and rounded shoulder); each molar was duplicated to fabricate 90 copings. A total of 180 copings generated 18 groups (n=10 for each finish line-coping material-luting agent combination). Luting agents tested included zinc phosphate, resin-modified glass ionomer (Fuji Plus), and resin composite cements (Panavia F). A metal frame was developed on which to screw the stainless steel model and a ceramic coping; the distance (mum) between 2 predetermined points was measured before and after cementation by a profile projector under a torquing force. A 4-way ANOVA with repeated measurements was performed to assess the influence of each factor in the vertical marginal discrepancy: 3 between-coping factors (finish line-coping material-luting agent) and 1 within-coping factor (before and after cementation) (alpha=.05).Results. Procera copings presented the lowest mean values (P<.05) of vertical marginal discrepancy before and after cementation (25/44 mum) when compared to Empress 2 (68/110 mum) and InCeram Alumina copings (57/117 mum), regardless of any combinations among all finish lines and luting agents tested.Conclusion. Considering each factor separately, the ceramic manufacturing technique appeared to be the most important factor tested for the definitive vertical discrepancy of all-ceramic copings, with lower mean values for Procera copings.

Inbreeding and its effects on some productive and reproductive traits in a Murrah buffalo herd

Data concerning genealogy, milk yield, lactation length, caving interval and lac at first calving, obtained from Murrah buffaloes on a farm In São Paulo state, Brazil, were studied in order to determine the presence of inbreeding in the herd and to evaluate its effects as well as the effects of some environmental factors on productive and reproductive traits. The mean inbreeding coefficient estimated for 2414 animals, born from 1965 to 1995 was 2.94%. of these animals, 47% mere considered inbred with a mean inbreeding coefficient equal 6.25%. Milk production, lactation length, calving interval and age at first calving averaged 1493.3 +/- 60.7 kg, 271.0 +/- 37.3 days, 385.0 +/- 53.4 days and 33.05 +/- 6.05 months, respectively. Among all four traits, only lactation length was affected by inbreeding.

Adhesive performance of dentin bonding agents applied in vivo and in vitro. Effect of intrapulpal pressure and dentin depth

The purpose of this study was to evaluate the influence of intrapulpal pressure and dentin depth on bond strengths of an etch-and-rinse and a self-etching bonding agent to dentin in vitro and in vivo. Twenty-four pairs of premolars were randomly divided into four groups (n = 6) according to the dentin bonding agent, Single Bond and Clearfil SE Bond, and intrapulpal pressure, null or positive. Each tooth of the pair was further designated to be treated in vivo or in vitro. The intrapulpal pressure was controlled in vivo by the delivery of local anesthetics containing or not a vasoconstrictor, while in vitro, it was achieved by keeping the teeth under hydrostatic pressure. Class I cavities were prepared and the dentin bonding agents were applied followed by incremental resin restoration. For the teeth treated in vitro, the same restorative procedures were performed after a 6 month-storage period. Beams with I mm 2 cross-sectional area were prepared and, microtensile tested. Clearfil SE Bond was not influenced by any of the variables of the study, while bond strengths produced in vitro were significatly higher for Single Bond. Overall, lower bond strengths were produced in deep dentin, which reached statistical significance when Single Bond was applied under physiological or simulated intrapulpal pressure. In conclusion, in vitro bonding may overestimate the immediate adhesive performance of more technique-sensitive dentin bonding systems. The impact of intrapulpal pressure on bond strength seems to be more adhesive dependent than dentin morphological characteristics related to depth. (C) 2007 Wiley Periodicals, Inc.

PHEIDOLE ANTS AS POTENTIAL BIOLOGICAL-CONTROL AGENTS OF THE BOLL-WEEVIL, ANTHONOMUS-GRANDIS (COL, CURCULIONIDAE), IN SOUTHEAST BRAZIL

This study evaluates the potential of ants as natural biological control agents of the boil weevil (Anthonomus grandis), during the between-season period, in South-east Brazil. Active adults of Anthonomus were experimentally distributed on the ground of the cotton field. Results show that 20% of the adult Anthonomus are attacked and removed by foraging ants. The native ant Pheidole oliveirai was by far the most efficient predator, accounting for 94% of the predation on Anthonomus. Recruited workers of P. oliveirai were usually very fast at transporting the weevils to their nests. The potential benefit of suppressing overwintering adult Anthonomus during the between-season period is mainly that of reducing the risk of high level infestations during the next cropping cycle.

Indole glucoalkaloids from Chimarrhis turbinata and their evaluation as antioxidant agents and acetylcholinesterase inhibitors

As part of our study on bioactive agents from Brazilian rainforest plants, two new glucoalkaloids, 3,4-dehydro-strictosidine (1) and 3,4-dehydro-strictosidinic acid (2), were isolated from Chimarrhis turbinata, along with seven known glucoalkaloids, cordifoline (3), strictosidinic acid (4), strictosidine (5), 5alpha-carboxystrictosidine (6), turbinatine (7), desoxycordifoline (8), and harman-3-carboxylic acid (9). The structures of the new alkaloids were established on the basis of comprehensive spectral analysis, mainly 1D and 2D NMR experiments, as well as high-resolution HRESIMS. Alkaloid 3 showed strong free-radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) as well as pronounced antioxidant activity evidenced by redox properties measured by ElCD-HPLC. Additionally, alkaloids 1-9 were submitted to TLC screening for acetylcholinesterase inhibitors. Both 7 and 8 were shown to be moderate acetylcholinesterase inhibitors at a concentration of 0.1 and 1.0 muM, respectively. In an in vitro rat brain assay, 7 showed moderate activity (IC50 1.86 muM), compared to the standard compound, galanthamine (IC50 0.92 muM).

Genetic damage in human peripheral lymphocytes exposed to antimicrobial endodontic agents

Objective. Formocresol, paramonochlorophenol, or calcium hydroxide have been widely used in dental practice to eradicate bacteria and consequently to produce root canal disinfection. Taking into consideration strong evidence for a relationship between DNA damage and carcinogenesis, the purpose of the present study was to evaluate the genotoxic effects of antimicrobial endodontic compounds in human peripheral lymphocytes by single-cell gel ( comet) assay. This technique detects DNA strand breaks in individual cells.Study design. A total of 10 mu L of the tested substance solution (formocreso1, paramonochlorofeno1, and calcium hydroxide at 100-mu g/mL concentration) was added to human peripheral lymphocytes from 10 volunteers for 1 hour at 37 degrees C. The negative control group was treated with vehicle control (PBS) for 1 hour at 37 degrees C, as well. For the positive control group, lymphocytes were exposed to hydrogen peroxide at 100 mu M during 5 minutes on ice.Results. No DNA breakage was detected after a treatment of peripheral lymphocytes by formocresol, paramonochlorophenol, or calcium hydroxide at 100 mu g/mL.Conclusions. In summary, our results indicate that exposure to formocresol, paramonochlorophenol, or calcium hydroxide may not be a factor that increases the level of DNA lesions in human peripheral lymphocytes as detected by single-cell gel (comet) assay.

Productive and reproductive performance in cattle infected with bovine leukosis virus

Economic losses caused by enzootic bovine leukosis (EBL) have been of interest since World War II, when the neoplastic form of EEL increased dramatically in Europe. Olson (1974) and House et al. (1975) ed that animals with lymphosarcoma caused by the bovine leukosis virus (BLV) had reduced milk yields. a less efficient reproductive performance and high veterinary costs and mortality rates, while many carcasses were rejected at slaughter. However, the actual impact of BLV infection in cattle without lymphosarcoma is not; clear. The purpose of the study reported here was to compare some productive and reproductive responses of cattle that were antibody-positive (BLV+) or negative (BLV-) for BLV.Holstein dairy cows in commercial dairy farms were used in this study. Blood samples were collected and subjected to BLV serological examination by the agar gel immunodiffusion test of Miller & van der Maaten (1976). Animals were then grouped as BLV+ or BLV- according to their serological response to the BLV antigen. Productive and reproductive histories were obtained from individual animal records and the following factors were considered: milk production, calving interval and birth rate. For milk production, we had the daily milk yields of 547 animals, and for calving interval the time between two successive parturitions for 444 cows. These values were examined by ANOVA and when this was significant a Student's t test was carried out for each age group. Birth rates, the percentage of animals that calved in 1 gear, were available for 557 animals and were examined with the Z-two proportion test. For all analyses, P < 0.05 was considered significant.

Effects of Antibacterial Agents on Dental Pulps of Monkeys Mechanically Exposed and Contaminated.

The purpose of this study was to compare the effectiveness of antibacterial agents and mineral trioxide aggregate in the healing of bacterial contaminated primate pulps. Study Design: The experiment required four adult male primates (Cebus opella) with 48 teeth prepared with buccal penetrartions into the pulpal tissues. The preparations (Cebus opella) with 48 teeth prepared with buccal penetrations into the exposed to cotton pellets soaked in a bacterial mixture consisting of microorganisms normally found in human pulpal abscesses obtained from the Endodontic Clinic of UNESP. Following bacterial inoculation (30 minute exposure), the pulpal tissue was immediately treated with either sterile saline, Cipro HC Otic solution (12), diluted Buckley formecresol solution (12) or Otosporin otic solution (12) for 5 minutes. After removal of the pellet, hemostasis was obtained and a ZOE base applied to the DFC treated pulps and the non-treated controls (12). After hemostasis, the other exposed pulps were covered with mineral trioxide aggregate (ProRoot). The pulpal bases were all covered with a RMGI (Fuji II LC). The tissue samples were collected at one day, two days, one week and over four weeks (34 days). Results: Following perfusion fixation, the samples were demineralized, sectioned, stained and histologically graded. After histologic analysis, presence of neutrophilic infiltrate and areas of hemorrhage with hyperemia were observed . The depth of the neutrophilic infiltrate depended on the agent or material used. The pupal tissue treated with Otic suspensions demonstrated significantly less inflammation (Kruskal Wallis non parametric analysis, H=9.595 with 1 degree of freedom; P=0.0223) than the formocresol and control groups. The hard tissue bridges formed over the exposure sites were more organized in the MTA treatment groups than in the control and ZOE groups (Kruskal Wallis non parametric analysis, H=18.291 with 1 degree of freedom; P=0.0004). Conclusions: Otic suspensions and MTA are effective in treating bacterial infected pulps and stimulate the production of a hard tissue bridge over the site of the exposure.

Effects of biofertilizer rates on the structural, morphogenetic and productive characteristics of Piata grass

The objective of this study was to investigate the influence of different levels of biofertilizers from cattle and swine manure on the structural, morphogenetic and productive characteristics of Brachiaria brizantha cv. Piata. The experiment was arranged in a completely randomized factorial design with split plots. The plots were defined by eight treatments: two biofertilizers (cattle and swine), four levels (0, 100, 200 and 300 kg N.ha(-1)) and subplots by four different cutting periods. The cutting for plant uniformity was performed at 45 days after sowing at 15 cm above the soil surface. The biofertilizeres were applied in a single level, after the cutting of plants, in rates of 0, 0.23 and 0.19, 0.45 and 0.38, 0.68 and 0.57 liters pot(-1) for the biofertilizers from cattle and swine manure, respectively. These rates were also equivalent to levels of 0, 100, 200 and 300 kg N.ha(-1). There was no significant difference between the types of biofertilizers as there was no interaction between them and the different levels, hence both biofertilizers could be applied without any loss of nutrient intake by the plants used in this experiment. There was a significant difference between the production of green and dry matter, the leaf appearance rate, phyllochron, leaf and pseudostem elongation rates, number of green leaves, final leaf length, number and weight of tillers, according to the increase of nitrogen rates, following linear prediction model. Effect of the cutting periods was also observed, once the plants harvested during the summer presented greater performance of structural and morphogenetic characteristics.

Thermoanalytical study of some anti-inflammatory analgesic agents

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