Effects of bovine Herpesvirus Type 5 on development of in vitro-produced bovine embryos

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

Evaluation of developmental changes in bovine in vitro produced embryos following exposure to bovine Herpesvirus type 5

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

ANALISE FAMILIAL DO COMPORTAMENTO IN VITRO DOS MACROFAGOS HUMANOS FRENTE AO MYCOBACTERIUM LEPRAE

The lysogenic capacity of human macrophages facing M. leprae in vitro may be dependent on an important genetic component. Although the familial aggregation of the trait is demonstrated, this is a necessary but not sufficient condition to prove genetic influence. The data do not fit some simple genetic models (autosomal dominant or incompletely dominant gene; dominant or recessive sex-linked gene). The results obtained are consistent with the hypothesis that the macrophages' lysogenic capacity is mainly due to a major gene with variable expressivity. This hypothesis may be too simple to account for the whole variability detected and therefore must be considered a working hypothesis.

Sister-chromatid exchanges in β-thalassaemic patients under conditions of in vivo and in vitro depletion of folic acid

In order to investigate the effect of folate depletion, lymphocyte sister-chromatid exchange (SCE) rates were compared among homozygous β-thalassaemic patients with low folic acid levels, heterozygous β-thalassaemic patients with normal folate levels and healthy persons with normal haemoglobin, in cultures with both normal and depleted folate conditions. Significantly higher SCE rates were found in homozygous patients in all assays, but the in vitro folate depletion did not induce an increase in SCE frequency in any group.

Evaluation of the in vitro susceptibility and emergence of mutants resistant to ciprofloxacin among multidrug resistant clinical isolates.

Two hundred and seventy-seven multidrug resistant clinical isolates [K. pneumoniae, (N = 87); E coli, (N = 30); Salmonella typhimurium (N = 100); P. aeruginosa, (N = 30); S. aureus, (N = 30)] from hospitalized patients specimens, were tested in vitro for sensitivity to Ciprofloxacin. Application of the disk diffusion test and determination of the minimal inhibitory concentration by the microdilution method indicated that, almost all isolates were sensitive to the drug. Overall, S. aureus and P. aeruginosa were the less sensitive organisms. Ciprofloxacin-resistant mutants occurred at frequencies of > or = 10(-5)/CFU.

Inhibition of demineralization in vitro around amalgam restorations

Objective: The purpose of this in vitro study was to evaluate some forms of preventing or avoiding demineralization within enamel cavity walls adjacent to amalgam restorations. Method and materials: Third molar teeth were sectioned to obtain 72 specimens, divided into one control and five experimental groups: amalgam only; varnish plus amalgam; acidulated phosphate fluoride plus amalgam; adhesive amalgam; glass-ionomer cement plus amalgam; control (amalgam only, not subjected to a demineralization challenge). The experimental groups were subjected to pH and thermal cycling and then submitted to enamel hardness determinations. Results: Significant differences between the treatment groups revealed that the bonded amalgam technique offered the best resistance to demineralization. The use of cavity varnish resulted in greater mineral loss than amalgam placed alone. Conclusion: The use of an adhesive system, glass-ionomer cement, or acidulated phosphate fluoride under amalgam restorations may interfere with development of secondary caries.

IL-12 and neutralization of endogenous IL-10 revert the in vitro antigen-specific cellular immunosuppression of paracoccidioidomycosis patients

Treatment of patients with paracoccidioidomycosis is still a challenge. Patients present defective lymphoproliferation and IFN-γ responses to the main Paracoccidioides brasiliensis antigen (gp43), which correlates with disease severity. Here, we demonstrated that the patients show also a defective synthesis of interleukin (IL)-12. Therefore, we attempted to revert this immune disfunction by adding IL-12 and neutralizing anti-IL-10 antibody to gp-43-stimulated peripheral blood mononuclear cell cultures. Both treatments increased IFN-γ secretion to levels observed with healthy sensitized individuals, but affected proliferation only modestly. When combined, the treatments further increased IFN-γ synthesis and cell proliferation. The addition of suboptimal concentrations of IL-2 also further increased the IL-12-mediated secretion of IFN-γ. Interestingly, the immune modulation was mostly antigen-specific, since the responses to Candida albicans' antigen were not affected. These results suggest that appropriate immune intervention with cytokines and/or anti-cytokines may help in the treatment of PCM. © 2002 Elsevier Science Ltd. All rights reserved.

Efeito dose-resposta de uma formulação de dentifrício com concentração reduzida de fluoreto - estudo in vitro.

The utilization of dentifrices with low fluorine concentration, for children under 6 years of age, has been suggested to reduce the risks of dental fluorosis. However, in order to have anticariogenic potential, the dentifrice should form loosely-bound fluorine (CaF2) on dental enamel. Considering that the formation of CaF2 is a function inversely related to pH, dentifrices with pH 5.5, with 275, 550 and 1,100 ppm F (NaF/silica) were developed in order to assess dose-response effects. A comparison between those dentifrices, a placebo product and the Crest toothpaste (positive control - standard) was carried out. Furthermore, the bioavailability of dentifrices, in terms of formation of total fluorine (TF), CaF2, and fluorapatite (FA) on human dental enamel, was evaluated. An ion-specific electrode was utilized for the determination of the dosage of fluorine. The results revealed that the dentifrice with 550 ppm F was more effective than both the placebo and the dentifrice with 275 ppm, presenting no difference in relation to the positive control (p > 0.05). A dose-effect correlation was observed as to the CaF2 formed. In conclusion, the modified formulation with 550 ppm F can be considered as effective as the standard dentifrice with 1,100 ppm, and its utilization would be safer with regard to dental fluorosis.

In vitro antifungal susceptibility of Candida spp. isolates from patients with chronic periodontitis and from control patients

Superinfection by Candida can be refractory to conventional periodontal treatments in specific situations, such as in immunocompromised patients. In these cases, the systemic therapy with antifungal drugs could be indicated. The aim of this study was to analyse antifungal susceptibility of Candida spp. strains isolated from chronic periodontitis patients and from control individuals. A total of 39 C. albicans isolates, 9 C. tropicalis, 2 C. glabrata and 5 Candida spp. from control individuals and 30 C. albicans, 3 C. tropicalis and 2 C. glabrata from periodontitis patients were tested. In the control group, 1 isolate of C. glabrata was resistant to ketoconazole and 1 Candida spp. was resistant to amphotericin B, ketoconazole and miconazole. Among the isolates of periodontitis group, 1 (3.33%) C. albicans isolate was resistant to flucytosine and ketoconazole. According to the obtained results, it could be concluded that fluconazole was the most effective drug against the several Candida species studied. There were not expressive differences in the susceptibility of isolates from periodontitis patients or from control individuals.

Fluoride does not induce DNA breakage in Chinese hamster ovary cells in vitro.

Fluoride has been widely used in dentistry because it is a specific and effective caries prophylactic agent. However, excess fluoride may represent a hazard to human health, especially by causing injury to genetic material. Genotoxicity tests represent an important part of cancer research to assess the risk of potential carcinogens. In the current study, the potential DNA damage associated with exposure to fluoride was assessed by the single cell gel (comet) assay in vitro. Chinese hamster ovary cells were exposed to sodium fluoride (NaF) at final concentration ranging from 7 to 100 micro/ml for 3 h, at 37 dgrees C. The results pointed out that NaF in all concentrations tested did not contribute to DNA damage as depicted by the mean tail moment and tail intensity. These findings are clinically important since they represent an important contribution to a correct evaluation of the potential health risk associated with the exposure to dental agents.

Influence of natural fruit juices in removing the smear layer from root surfaces - An in vitro study

Certain elements of a patient's diet may be associated with dentin hypersensitivity. The intent of this study was to evaluate the degree of removal of the smear layer from dentin surfaces by various fruit juices. A smear layer was created on extracted human teeth by manual scaling. The roots were reduced and distributed into 8 experimental groups. Distilled water was the negative control. The juices were applied by 2 methods: topical application and topical application with friction. Specimens were photomicrographed and graded according to an index of smear layer removal. With topical application, all but 2 of the tested substances resulted in significantly greater removal of the smear layer and opening of dentinal tubules than was the case with the negative control (p = 0.05); the exceptions were Gala apple and Italian grape juices, which were no different from the control. For the active application (with friction), most substances removed more smear layer than the control (p < 0.05); Gala apple, Italian grape and orange juices were similar to the control. For each of the tested substances, removal of the smear layer did not differ with the method of application (topical vs. friction; p > 0.05). It is concluded that natural fruit juices can remove the smear layer from dentin surfaces, and the efficacy of this removal varies with the type of juice. © J Can Dent Assoc 2004.

Magnetic images of pharmaceutical dosage forms in the human gastrointestinal tract

Oral administration with solid dosage forms is a common route in the drug therapy widely used. The drug release by the disintegration process occurs in several gastrointestinal tract (GIT) regions. AC Biosusceptometry (ACB) was originally proposal to characterize the disintegration process of tablets in vitro and in the human stomach, through changes in magnetic signals. The aim of this work was to employ a multisensor ACB system to monitoring magnetic tablets and capsules in the human GIT and to obtain the magnetic images of the disintegration process. The ACB showed accuracy to quantify the gastric residence time, the intestinal transit time and the magnetic images allowed to visualize the disintegration of magnetic formulations in the GIT. The ACB is a non-invasive, radiation free technique, completely safe and harmless to the volunteers and had demonstrated potential to evaluate pharmaceutical dosage forms in the human gastrointestinal tract. © 2005 IEEE.

In vitro evaluation of apical microleakage using different root-end filling materials

The objective of this study was to evaluate the apical leakage of retrograde cavities filled with Portland Cement (Concrebrás S/A-MG-Brazil), ProRoot MTA™ (Dentsply International, Johnson City, TN, USA) and Sealapex (Kerr Corporation, Orange, California, USA) with addition of zinc oxide (Odahcam Herpo Produtos Dentários Ltda, Rio de Janeiro, RJ, Brazil). Forty-two extracted single-rooted human teeth were decoronated and used for this study. The root canals were instrumented at 1.0mm short of the apical foramen using the step-back technique to an apical ISO size 60. The roots were obturated with gutta-percha points and sealer Sealapex (Kerr Corporation-USA) and then 3mm of each root apex was sectioned at a 90° angle. Ultrasonic retrograde preparation was performed with a diamond tip to 3mm depth and the roots were randomly divided into 3 groups according to the filling material: G1-Portland, G2-ProRoot MTA, G3- Sealapex zinc oxide-added cement. The root surfaces were covered with nail varnish up to 2mm from the apical foramen, immersed in simulated tissue fluid for 30 days, and then immersed in 0.2% Rhodamine B solution for 24 hours for evaluation of marginal leakage. The results showed mean leakage of 0.75, 0.35 and 0.35 for groups 1, 2 and 3, respectively; however, Kruskal-Wallis test revealed that there was no statistically significant difference among the results (p>0.05).

In vitro cytotoxic and non-genotoxic effects of gutta-percha solvents on mouse lymphoma cells by single cell gel (comet) assay

Chloroform and eucalyptol are widely used in clinical dentistry as gutta-percha solvents. However, these compounds may represent a hazard to human health, especially by causing injury to genetic apparatus and/or inducing cellular death. In this study, the genotoxic and cytotoxic potentials associated with exposure to chloroform and eucalyptol were assessed on mouse lymphoma cells in vitro by the single cell gel (comet) assay and trypan blue exclusion test, respectively. Both gutta-percha solvents proved to be cytotoxic at the same levels in concentrations of 2.5, 5 and 10 μL/mL (p<0.05). On the other hand, neither of the solvents induced DNA breakage. Taken together, these results suggest that although both tested compounds (chloroform and eucalyptol) are strong cytotoxicants, it seems that they are not likely to increase the level of DNA damage on mammalian cells.