The amount of digesta flowing to the duodenum is a relevant measurement for the evaluation of nutrient supply to ruminants, which is usually estimated in animals fitted with a duodenal T-type cannula ...using internal or external markers. This study evaluated acid detergent fiber (ADF) compared with external (C32n-alkane) and internal sulfuric acid lignin (ADL) and n-alkanes C31 and C33 markers for estimating duodenal flow and(or) ruminal digestibility of dry matter (DM) in cattle and sheep. In the first assay, 4 duodenally cannulated Holstein steers housed in metabolism cages, dosed with C32n-alkane, and fed Avena strigosa plus concentrate and increasing levels of tannin extract to reduce ruminal digestibility, were used in a Latin square design. The mobile-bag technique was used to measure the intestinal disappearance of ADL and ADF from forage (Avena strigosa, Pennisetum purpureum, Cynodon dactylon, and Medicago sativa) and concentrate (corn grain, soybean meal, and sunflower meal) samples that were previously incubated in the rumen of additional fistulated steer for 12, 24, 36, or 48h. The ADF concentration in residues recovered in the feces was strongly related to the ADF concentration in residues at the duodenum (R2=0.93, standard deviation=30.0, n=901). This relationship showed a lower precision for ADL fraction (R2=0.88, standard deviation=12.6, n=590). In a second assay, duodenal flow and ruminal DM digestibility were calculated from the duodenal and fecal concentration of either marker. We observed a significant effect of marker type on ruminal DM digestibility values, and the effect of tannin treatments was observed only when ADF or ADL was used as the marker. The lowest residual error was obtained for ADF. Ruminal DM digestibility was, on average, higher for C31 and C33n-alkanes, and the use of dosed C32n-alkane resulted in a negative value. In the third assay, a data set of 235 individual observations was compiled from digestibility trials to compare ADF and ADL as markers for estimating duodenal digesta flow in wethers (n=204) and cattle (n=31). We observed a strong relationship between markers (R2=0.84 in sheep and R2=0.88 in cattle), but variance analysis within trials indicated that ADF was more precise than ADL. In conclusion, in digestibility trials in which fecal output was measured and spot samples of the duodenal digesta were obtained, duodenal flow and ruminal digestibility of the DM may be estimated from the relationship between the ADF concentration in feces and that in the duodenal digesta of ruminants.
Silva EJNL, Herrera DR, Almeida JFA, Ferraz CCR, GomesBPFA, Zaia AA. Evaluation of cytotoxicity and up‐regulation of gelatinases in fibroblast cells by three root repair materials. International ...Endodontic Journal, 45, 815–820, 2012.
Aim To investigate the effects of root repair materials on the cytotoxicity and gelatinolytic activity of matrix metalloproteinases (MMPs) in 3T3 fibroblasts.
Methodology Fibroblasts (3T3, 3 × 105 cells per well) were incubated with elutes of calcium hydroxide (Biodinâmica, Ibiporã, PR, Brazil), EndoBinder (Binderware, São Carlos, SP, Brazil) and mineral trioxide aggregate (MTA) (Angelus, Londrina, PR, Brazil) for 24 h. The cytotoxicity of all root repair materials was determined using the MTT assay. Supernatants of cell cultures incubated with materials were collected after 24 h to determine the levels of MMP‐2 and MMP‐9 gelatinolytic activity by gelatin zymography. Data were analysed using anova and Tukey’s test.
Results Cells secreted MMP‐2 after 24 h with calcium hydroxide inducing significantly greater MMP‐2 expression in relation to the control and the other root repair materials (P < 0.05). The cytotoxicity results revealed that there was no significant difference in the cell viability of MTA, EndoBinder and the control group. However, there was a significantly reduced cell viability of 3T3 fibroblasts in association with calcium hydroxide (P < 0.05).
Conclusions Calcium hydroxide was associated with significantly less cell viability when compared with EndoBinder and MTA. All materials had gelatinolytic activity for MMP‐2 with calcium hydroxide being associated with the greatest activity.
β-Glucosidases are important enzymes with significant prospects in the industrial biotechnology, including their use in biomass hydrolysis for bioethanol production. In this study, the use of canola ...meal as carbon source for β-glucosidase production by a Trichoderma viride strain in submerged fermentation was evaluated by applying central composite design and response surface methodology to optimize the production process. This statistical approach was also used to improve the passion fruit peel hydrolysis by T. viride crude extract. The model developed 3.6-fold increased β-glucosidase activity. The culture conditions that resulted in the highest β-glucosidase levels were a substrate concentration of 2.9 %, pH of medium 4.2 and cultivation time of 206 h. The β-glucosidases produced under optimal conditions showed attractive properties for industrial applications, such as activity at high temperatures and stability at 55 °C and over a wide pH range. In addition, the enzymatic hydrolysis of passion fruit peel by T. viride crude extract was very promising, resulting in glucose yields of 66.4 %. This study, therefore, presents canola meal as an inexpensive and attractive substrate for the production of microbial β-glucosidases.
Background. Exercise is increasingly becoming recognized as an important adjunct to medications in the clinical management of Parkinson’s disease (PD). Boxing and sensory exercise have shown ...immediate benefits, but whether they continue beyond program completion is unknown. This study aimed to investigate the effects of boxing and sensory training on motor symptoms of PD, and whether these benefits remain upon completion of the intervention. Methods. In this 20-week double-blinded randomized controlled trial, 40 participants with idiopathic PD were randomized into 2 treatment groups, (n = 20) boxing or (n = 20) sensory exercise. Participants completed 10 weeks of intervention. Motor symptoms were assessed at (week 0, 10, and 20) using the Unified Parkinson’s Disease Rating Scale (UPDRS-III). Data were analyzed using SPSS, and repeated-measures ANOVA was conducted. Results. A significant interaction effect between groups and time were observed F(1, 39) = 4.566, P = .036, where the sensory group improved in comparison to the boxing group. Post hoc analysis revealed that in comparison to boxing, the effects of exercise did not wear off at washout (week 20) P < .006. Conclusion. Future rehabilitation research should incorporate similar measures to explore whether effects of exercise wear off post intervention.
One of the most crucial events during infection with the dimorphic fungus Paracoccidioides brasiliensis is adhesion to pulmonary epithelial cells, a pivotal step in the establishment of disease. In ...this study, we have evaluated the relevance of a 32-kDa protein, a putative adhesion member of the haloacid dehalogenase (HAD) superfamily of hydrolases, in the virulence of this fungus. Protein sequence analyses have supported the inclusion of PbHad32p as a hydrolase and have revealed a conserved protein only among fungal dimorphic and filamentous pathogens that are closely phylogenetically related. To evaluate its role during the host-pathogen interaction, we have generated mitotically stable P. brasiliensis HAD32 (PbHAD32) antisense RNA (aRNA) strains with consistently reduced gene expression. Knockdown of PbHAD32 did not alter cell vitality or viability but induced morphological alterations in yeast cells. Moreover, yeast cells with reduced PbHAD32 expression were significantly affected in their capacity to adhere to human epithelial cells and presented decreased virulence in a mouse model of infection. These data support the hypothesis that PbHad32p binds to extracellular matrix (ECM) proteins and modulates the initial immune response for evasion of host defenses. Our findings point to PbHAD32 as a novel virulence factor active during the initial interaction with host cells in P. brasiliensis.
Aim
To analyse the influence of exposure of mineral trioxide aggregate (MTA) with and without calcium chloride (CaCl2) to phosphate‐buffered saline (PBS) on the push‐out bond strength, over different ...experimental periods.
Methodology
One hundred and twenty dentine discs with standardized cavities were filled with MTA with and without 10% CaCl2. The specimens were randomly divided as follows (n = 30): (G1) MTA in contact with a moistened cotton pellet, (G2) MTA immersed in PBS, (G3) MTA + CaCl2 in contact with a moistened cotton pellet and (G4) MTA + CaCl2 immersed in PBS. The samples were stored for 3, 28 and 60 days. The bond strengths were measured with the Instron Testing machine. Data were analysed using the three‐way anova and Tukey test (P < 0.05).
Results
In general, the samples of MTA with and without CaCl2, exposed to PBS, had higher bond strength values in all study periods (P < 0.05). Analysis of the influence of addition of CaCl2 to MTA (G1 × G3) evidenced significant differences in bond strength in the different periods (P < 0.05).
Conclusions
The exposure of MTA to PBS positively influenced the push‐out bond strength, whereas the addition of CaCl2 had a negative influence.
In the mammalian testis, Leydig cells are primarily responsible for steroidogenesis. In adult stallions, the major endocrine products of Leydig cells include testosterone and estrogens. ...3β-hydroxysteroid dehydrogenase/Δ⁵-Δ⁴-isomerase (3βHSD) and 17α-hydroxylase/17,20-lyase (P450c17) are two key steroidogenic enzymes that regulate testosterone synthesis. Androgens produced by P450c17 serve as substrate for estrogen synthesis. The aim of this study was to investigate localization of the steroidogenic enzymes P450c17, 3βHSD, and P450arom and to determine changes in expression during development in the prepubertal, postpubertal, and adult equine testis based upon immunohistochemistry (IHC) and real-time quantitative PCR. Based on IHC, 3βHSD immunolabeling was observed within seminiferous tubules of prepubertal testes and decreased after puberty. On the other hand, immunolabeling of 3βHSD was very weak or absent in immature Leydig cells of prepubertal testes and increased after puberty. HSD3B1 (3βHSD gene) mRNA expression was higher in adult testes compared with prepubertal (P=0.0001) and postpubertal testes (P=0.0041). P450c17 immunolabeling was observed in small clusters of immature Leydig cells in prepubertal testes and increased after puberty. CYP17 (P450c17 gene) mRNA expression was higher in adult testes compared with prepubertal (P=0.030) and postpubertal testes (P=0.0318). A weak P450arom immunolabel was observed in immature Leydig cells of prepubertal testes and increased after puberty. Similarly, CYP19 (P450arom gene) mRNA expression was higher in adult testes compared with prepubertal (P=0.0001) and postpubertal (P=0.0001) testes. In conclusion, Leydig cells are the primary cell type responsible for androgen and estrogen production in the equine testis.
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