Modulatory effect of Byrsonima basiloba extracts on the mutagenicity of certain direct and indirect-acting mutagens in Salmonella typhimurium assays

Byrsonima basiloba A. Juss. species is a native arboreal type from the Brazilian cerrado (tropical American savanna), and the local population uses it to treat diseases, such as diarrhea and gastric ulcer. It belongs to the Malpighiaceae family, and it is commonly known as murici. Considering the popular use of B. basiloba derivatives and the lack of pharmacological potential studies regarding this vegetal species, the mutagenic and antimutagenic effect of methanol (MeOH) and chloroform extracts were evaluated by the Ames test, using strains TA97a, TA98, TA100, and TA102 of Salmonella typhimurium. No mutagenic activity was observed in any of the extracts. To evaluate the antimutagenic potential, direct and indirect mutagenic agents were used: 4 nitro-o-phenylenediamine, sodium azide, mitomycin C, aflatoxin B1, benzo[a]pyrene, and hydrogen peroxide. Both the extracts evaluated showed antimutagenic activity, but the highest value of inhibition level (89%) was obtained with the MeOH extract and strain TA100 in the presence of aflatoxin B1. Phytochemical analysis of the extracts revealed the presence of n-alkanes, lupeol, ursolic and oleanolic acid, (+)-catechin, quercetin-3-O-α-L-arabinopyranoside, gallic acid, methyl gallate, amentoflavone, quercetin, quercetin-3-O-(2″-O-galloyl)-β-D- galactopyranoside, and quercetin-3-O-(2″-O-galloyl)-α-L- arabinopyranoside. © 2008 Mary Ann Liebert, Inc.

Direct and indirect connections between cochlear root neurons and facial motor neurons: Pathways underlying the acoustic pinna reflex in the albino rat

Cochlear root neurons (CRNs) are involved in the acoustic startle reflex, which is widely used in behavioral models of sensorimotor integration. A short-latency component of this reflex, the auricular reflex, promotes pinna movements in response to unexpected loud sounds. However, the pathway involved in the auricular component of the startle reflex is not well understood. We hypothesized that the auricular reflex is mediated by direct and indirect inputs from CRNs to the motoneurons responsible for pinna movement, which are located in the medial subnucleus of the facial motor nucleus (Mot7). To assess whether there is a direct connection between CRNs and auricular motoneurons in the rat, two neuronal tracers were used in conjunction: biotinylated dextran amine, which was injected into the cochlear nerve root, and Fluoro-Gold, which was injected into the levator auris longus muscle. Under light microscopy, close appositions were observed between axon terminals of CRNs and auricular motoneurons. The presence of direct synaptic contact was confirmed at the ultrastructural level. To confirm the indirect connection, biotinylated dextran amine was injected into the auditory-responsive portion of the caudal pontine reticular nucleus, which receives direct input from CRNs. The results confirm that the caudal pontine reticular nucleus also targets the Mot7 and that its terminals are concentrated in the medial subnucleus. Therefore, it is likely that CRNs innervate auricular motoneurons both directly and indirectly, suggesting that these connections participate in the rapid auricular reflex that accompanies the acoustic startle reflex. © 2008 Wiley-Liss, Inc.

In vitro study of shear bond strength in direct bonding of orthodontic molar tubes

Objective: Although direct bonding takes up less clinical time and ensures increased preservation of gingival health, the banding of molar teeth is still widespread nowadays. It would therefore be convenient to devise methods capable of increasing the efficiency of this procedure, notably for teeth subjected to substantial masticatory impact, such as molars. This study was conducted with the purpose of evaluating whether direct bonding would benefit from the application of an additional layer of resin to the occlusal surfaces of the tube/tooth interface. Methods: A sample of 40 mandibular third molars was selected and randomly divided into two groups: Group 1 - Conventional direct bonding, followed by the application of a layer of resin to the occlusal surfaces of the tube/tooth interface, and Group 2 - Conventional direct bonding. Shear bond strength was tested 24 hours after bonding with the aid of a universal testing machine operating at a speed of 0.5mm/min. The results were analyzed using the independent t-test. Results: The shear bond strength tests yielded the following mean values: 17.08 MPa for Group 1 and 12.60 MPa for Group 2. Group 1 showed higher statistically significant shear bond strength than Group 2. Conclusions: The application of an additional layer of resin to the occlusal surfaces of the tube/tooth interface was found to enhance bond strength quality of orthodontic buccal tubes bonded directly to molar teeth.

The influence of the flexibility of the chair seat on pressure peak and distribution of the contact area in individuals with cerebral palsy during the execution of a task

This study aimed to determine the influence of flexibility of the chair seat surface on the pressure peak and on the contact area during the execution of a task of handling an object on the seated position by individuals with spastic cerebral palsy. Ten individuals of both genders with diagnosis of spastic cerebral palsy, who had some control to voluntarily move the body and the upper limbs, participated in this study. Quantification of data was carried out in two experimental situations: (1) execution of a task of fitting with upper limbs, and with the individual placed on an adapted canvas seat; (2) execution of a task of fitting with the participant positioned on an adapted wooden seat. Data obtained were submitted to a non-parametric and descriptive statistical analysis using the Wilcoxon test. Results indicated that the use of canvas seat increased the contact area and decreased the pressure peak and the medio-lateral displacement of centre pressure on the seated posture. © 2011 Informa UK, Ltd.

Chemoprotective activity of the isoflavones, genistein and daidzein on mutagenicity induced by direct and indirect mutagens in cultured HTC cells

Isoflavones are phenolic compounds widely distributed in plants and found in a high percentage in soybeans. They have important biological properties and are regarded as potential chemopreventive agents. The aim of this study was to verify the preventive effect of two soy isoflavones (genistein and daidzein) by a micronucleus assay, analysis of GST activity, and real-time RT-PCR analysis of GSTa2 gene expression. Mutagens of direct (doxorubicin) and indirect (2-aminoanthracene) DNA damage were used. Hepatoma cells (HTC) were treated with genistein or daidzein for 26 h at noncytotoxic concentrations; 10 μM when alone, and 0.1, 1.0 and 10 μM when combined with genotoxic agents. The micronucleus test demonstrated that both isoflavones alone had no genotoxic effect. Genistein showed antimutagenic effects at 10 μM with both direct and indirect DNA damage agents. On phase II enzyme regulation, the current study indicated an increase in total cytoplasmic GST activity in response to genistein and daidzein at 10 μM supplementation. However, the mRNA levels of GSTa2 isozymes were not differentially modulated by genistein or daidzein. The results point to an in vitro antimutagenic activity of genistein against direct and indirect DNA damage-induced mutagenicity. © 2012 Springer Science+Business Media B.V.

Ethopharmacological evaluation of the rat exposure test: A prey-predator interaction test

The rat exposure test (RET) is a prey (mouse)-predator (rat) situation that activates brain defensive areas and elicits hormonal and defensive behavior in the mouse. Here, we investigated possible correlations between the spatiotemporal [time spent in protected (home chamber and tunnel) and unprotected (surface) compartments and frequency of entries into the three compartments] and ethological [e.g., duration of protected and unprotected stretched-attend postures (SAP), duration of contact with the rat's compartment] measures (Experiment 1). Secondly, we investigated the effects of systemic treatment with pro- or anti-aversive drugs on the behavior that emerged from the factor analysis (Experiment 2). The effects of chronic (21 days) imipramine and fluoxetine on defensive behavior were also investigated (Experiment 3). Exp. 1 revealed that the time in the protected compartment, protected SAP and rat contacts loaded on factor 1 (defensive behavior), while the total entries and unprotected SAP loaded on factor 2 (locomotor activity). Exp. 2 showed that alprazolam (but not diazepam) selectively changed the defensive factor. Caffeine produced a mild proaversive-like effect, whereas yohimbine only decreased locomotor activity (total entries). Fluoxetine (but not imipramine) produced a weak proaversive-like effect. 5-HT1A/5-HT2 receptor ligands did not change any behavioral measure. In Exp. 3, chronic fluoxetine (but not imipramine) attenuated the defensive behavior factor without changing locomotion. Given that the defensive factor was sensitive to drugs known to attenuate (alprazolam and chronic fluoxetine) and induce (caffeine) panic attack, we suggest the RET as a useful test to assess the effects of panicolytic and panicogenic drugs. © 2012 Elsevier B.V.

Genetic parameters for test-day milk yield, 305-day milk yield, and lactation length in Guzerat cows

Milk production in tropical environments requires the use of crossbreeding systems including breeds well adapted to harsh conditions, but with lower productivities when compared to specialized breeds. Besides the genetic improvement for milk production, lactation lengths also need to be studied for most of these breeds. Accordingly, genetic parameters were estimated for 305-day cumulative milk yield (MY305), test-day milk yield (TDMY), and lactation length (LL) using information from the first lactations of 2816 Guzerat cows selected for milk production in 28 herds in Brazil. Contemporary groups were defined as herd, year and season of the test for TDMY, and as herd, year and season of calving for MY305 and LL. Variance components were estimated with the restricted maximum likelihood method under a multi-trait animal model. Heritabilities estimated for TDMY ranged from 0.16 to 0.24, and were 0.24 and 0.12 for MY305 and LL, respectively. Genetic correlations were high and positive, ranging from 0.51 to 0.99 among TDMY records, from 0.81 to 0.98 between each TDMY and MY305, and from 0.71 to 0.94 between each TDMY and LL. Genetic parameters obtained in this study indicated the possibility of using test-day records for the prediction of breeding values for milk yield in this population of the Guzerat breed. The use of TDMY as selection criteria would result in indirect gains in MY305 and LL. However, the highest response to selection for MY305 would be obtained by direct selection for this trait. © 2012 Elsevier B.V.

Deproteinized bovine bone mineral particles and osseointegration of implants without primary bone contact: An experimental study in dogs

Objectives: To evaluate the influence on osseointegration of Deproteinized bovine bone mineral (DBBM) particles used to fill defects of at least 1 mm around implants having no primary contact with bone. Material and methods: Premolars and first molars were extracted bilaterally from the mandible of six Labrador dogs. After 3 months of healing, mucoperiosteal full-thickness flaps were elevated, and one recipient site was prepared in the molar region of each hemi-mandible to place implants. These were installed with a deliberate circumferential and periapical space to the bone walls of 1.2 mm. All implants were stabilized with passive fixation plates to maintain the implants in situ and without any contact with the implant bed. The control sites were left to be filled with coagulum, while at the test sites, the residual gap was filled with DBBM. After 3 months of submerged healing, the animals were sacrificed. Ground sections were prepared and analyzed histomorphometrically. Results: Mineralized bone-to-implant contact was 4.0% and 3.9% for control and test sites, respectively. The width of the residual defects was 0.48 mm and 0.88 mm at the control and test sites, respectively. The percentage of implant surface covered by a layer of dense connective tissue of 0.12 mm of width on average was 84.9% and 88.5% at the control and test sites, respectively. Conclusion: A minor and not predictable degree of contact or distance osteogenesis was obtained on the implant surface when primary contact of the implant surface with the implant bed had deliberately been avoided. DBBM grafting of the artificial gap did not favor osseointegration. Neither did it enhance the ability to bridge the gap with newly formed bone in an artificial defect wider than 1 mm. © 2013 John Wiley & Sons A/S.

Effect of light curing modes on mechanical properties of direct and indirect composites

Objective. To evaluate the degree of conversion (DC), flexural strength (FS) and Knoop microhardness (KHN) of direct and indirect composite resins polymerized with different curing systems. Materials and methods. Specimens of direct (Z250, 3M/Espe) and indirect (Sinfony, 3M/Espe) restorative materials were made and polymerized using two light curing units: XL2500 (3M/Espe) and Visio system (3M/Espe). Absorption spectra of both composites were obtained on a FTIR spectrometer in order to calculate the DC. FS was evaluated in a universal testing machine and surface microhardness was performed in a microhardness tester (50gf/15s). DC, FS and KHN data were submitted to two-way ANOVA and Tukey's test (α = 0.05). Results. Z250 showed higher DC, FS and KHN compared with Sinfony when the polymerization was carried out with XL2500 (p < 0.05). However, there is no statistical difference in DC between the materials when Visio was used (p > 0.05). Visio showed higher DC and KHN for Z250 and Sinfony than the values obtained using XL2500 light curing (p < 0.05). For FS, no significant difference between curing units was found (p > 0.05). Conclusion. Even though the Visio system could increase DC and KHN for some direct and indirect composites, compared with the conventional halogen curing unit, a high number of monomers did not undergo conversion during the polymerization. © 2013 Informa Healthcare.

Noradrenergic neurotransmission within the bed nucleus of the stria terminalis modulates the retention of immobility in the rat forced swimming test

The bed nucleus of the stria terminalis (BNST) is a limbic structure that has a direct influence on the autonomic, neuroendocrine, and behavioral responses to stress. It was recently reported that reversible inactivation of synaptic transmission within this structure causes antidepressant-like effects, indicating that activation of the BNST during stressful situations would facilitate the development of behavioral changes related to the neurobiology of depression. Moreover, noradrenergic neurotransmission is abundant in the BNST and has an important role in the regulation of emotional processes related to the stress response. Thus, this study aimed to test the hypothesis that activation of adrenoceptors within the BNST facilitates the development of behavioral consequences of stress. To investigate this hypothesis, male Wistar rats were stressed (forced swimming, 15 min) and 24 h later received intra-BNST injections of vehicle, WB4101, RX821002, CGP20712, or ICI118,551, which are selective α1, α2, β1, and β2 adrenoceptor antagonists, respectively, 10 min before a 5-min forced swimming test. It was observed that administration of WB4101 (10 and 15 nmol), CGP20712 (5 and 10 nmol), or ICI118,551 (5 nmol) into the BNST reduced the immobility time of rats subjected to forced swimming test, indicating an antidepressant-like effect. These findings suggest that activation of α1, β1, and β2 adrenoceptors in the BNST could be involved in the development of the behavioral consequences of stress. © 2013 Wolters Kluwer Health | Lippincott Williams & Wilkins.

Comparação de técnicas no diagnóstico da infecção leptospírica em ovinos e implicações na saúde pública

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

Influência de Lactobacillus acidophilus sobre biofilme formado por Candida albicans in vitro e infecção em modelo de invertebrado

Pós-graduação em Biopatologia Bucal - ICT

Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR

Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

Soroprevalência de testes rápidos (ML Flow) em casos de hanseníase e contatos intradomiciliares em municípios endêmicos do Pará

A hanseníase, doença milenar, ainda constitui um grande desafio para a saúde pública, inclusive no que se refere a diagnóstico precoce e vigilância em saúde. Com objetivo de estimar a soroprevalência de anticorpos antiPGL-1 através do teste rápido ML-Flow em casos de hanseníase e seus contatos intradomiciliares de municípios endêmicos do Pará, realizou-se um estudo transversal incluindo 73 casos novos de hanseníase e 135 contatos intradomiciliares, selecionados no período de abril de 2011 a janeiro de 2012, nas unidades de referência para tratamento de hanseníase nos municípios de Belém, Marituba, Igarapé-Açú e Santarém. Os resultados demonstraram uma prevalência de 14,8/10.000hab de casos de hanseníase entre os contatos examinados. A soropositividade do ML Flow nos casos índices foi de 53,42% em pacientes multibacilares e 13,33% nos contatos intradomiciliares. Houve associação direta de positividade do ML Flow nos contatos intradomiciliares com o índice baciloscópico do caso índice. Não houve associação direta do tempo de convivência e a consanguineidade do caso índice em relação ao teste ML Flow. Houve associação entre a positividade do teste ML Flow e a realização da vacina BCG entre os contatos. Estes resultados indicam que a introdução do teste ML Flow poderia servir como instrumento auxiliar no monitoramento dos casos e seus contatos, tornando-se de grande relevância na vigilância epidemiológica da hanseníase.

Effect of serum sample inactivation on the performance of latex agglutination test for paracoccidioidomycosis serodiagnosis

Paracoccidioidomycosis is diagnosed from the direct observation of the causative agent, but serology can facilitate and decrease the time required for diagnosis. The objective of this study was to determine the influence of serum sample inactivation on the performance of the latex agglutination test (LAT) for detecting antibodies against Paracoccidioides brasiliensis. The sensitivity of LAT from inactivated or non-inactivated samples was 73% and 83%, respectively and the LAT selectivity was 79% and 90%, respectively. The LAT evaluated here was no more specific than the double-immunodiffusion assay. We suggest the investigation of other methods for improving the LAT, such as the use of deglycosylated antigen.