Fertility-associated proteins in Nelore bull sperm membranes

The present study was undertaken to evaluate the protein composition of the sperm membranes (SM) of Nelore bulls, assessing protein markers associated with bull fertility, and whether these markers can be used for predicting bull fertility. Samples were obtained of 20 Nelore bulls, with fertility ranked and divided into three groups (greater, normal and least). To rank the bull's fertility weighted classification was used (according to the number of pregnant cows, number of AI cows and number of herds, considering three different breeding seasons), using the PROC GENMOD as a statistical model, with 99% significance. A total of 7897 Nelore cows, randomly distributed among 28 different farms, were considered in the statistical analyses. The bulls were divided into three fertility groups (pregnancy rates): greater (%F > 80), normal (79 <%F > 71) and least (< 68%F) with 3, 13 and 4 bulls, respectively. Two-dimensional gel electrophoresis (2DE) of sperm membranes indicated in 27 spots (SM40, SM53, SM69, SM93, SM102, SM111, SM137, SM138, SM189, SM196, SM201, SM202, SM204, SM225, SM236, SM237, SM239, SM241, SM246, SM247, SM275, SM283, SM342, SM346, SM355, SM372, SM391) was prevalent in the higher fertility group, and just one spot (SM244) was prevalent in the lower fertility group. Spots SM244 and SM239 had their identification defined by PMF/MALDI-MS, as BSP-A3 and aSFP, respectively. Both these proteins showed a great potential for predicting bull's fertility. The amount of aSFP was 8.5 times greater in the sperm membrane protein profile of the higher fertility groups of Nelore bulls. Besides that, the BSP-A3 was 2.5 times greater in the lower fertility group. For the other spots potentially associated with fertility not yet identified, additional tests will be necessary, but it is clear that the 2D electrophoresis of the sperm membrane can be used for a new approach to predict Nelore bull fertility. (c) 2005 Elsevier B.V. All rights reserved.

SPINDLE MEMBRANES DURING SPERMATOGENESIS IN DERMATOBIA-HOMINIS (DIPTERA, CUTEREBRIDAE)

During the mitotic and meiotic division in Dermatobia hominis spermatogenesis, the nuclear envelope is fragmented and membranes appear around the spindle. The membranes surrounding the mitotic spindle are formed by two layers of cisterns. The membranes of the meiotic spindle consist in at least 3 or 4 layers of long smooth cisterns which isolate the spindle from the remaining cytoplasm. The presence of this kind of membranes during meiosis seems to be usual in insect male germ cell.

Collagen membranes at immediate implants: a histomorphometric study in dogs

AimTo evaluate the influence of resorbable membranes on hard tissue alterations and osseointegration at implants placed into extraction sockets in a dog model.Material and methodsIn the mandibular premolar region, implants were installed immediately into the extraction sockets of six Labrador dogs. Collagen-resorbable membranes were placed at the test sites, while the control sites were left uncovered. Implants were intended to heal in a submerged mode. After 4 months of healing, the animals were sacrificed, and ground sections were obtained for histomorphometric evaluation.ResultsAfter 4 months of healing, a control implant was not integrated (n=5). Both at the test and at the control sites, bone resorption occurred. While the most coronal bone-to-implant contact was similar between the test and the control sites, the alveolar bone crest outline was maintained to a higher degree at the buccal aspect of the test sites (loss: 1.7 mm) compared with the control sites (loss: 2.2 mm).ConclusionsThe use of collagen-resorbable membranes at implants immediately placed into extraction sockets contributed to a partial (23%) preservation of the buccal outline of the alveolar process.To cite this article:Caneva M, Botticelli D, Salata LA, Souza SLS, Carvalho Cardoso L, Lang NP. Collagen membranes at immediate implants: a histomorphometric study in dogs.Clin. Oral Impl. Res. 21, 2010; 891-897.doi: 10.1111/j.1600-0501.2010.01946.x.

Changes in norepinephrine and epinephrine concentrations in adrenal gland of the rats submitted to acute immobilization stress

Catecholamines act as neurotransmitters and hormones. Studies conducted to understand the synthesis and metabolism of these monoamines during stress have been the main concern of many authors. This work proposes to investigate the time course of changes in epinephrine and norepinephrine concentration in adrenal gland obtained from rats submitted to acute immobilization stress. The results of the present study indicate that acute immobilization stress during 5 and 15min did not provoke changes in epinephrine and norepinephrine concentrations in adrenal gland in relation to the control group. Such results are justified due to the short time of the stress, showing that the stress did not provoke physiological alteration. The epinephrine and norepinephrine concentrations in adrenal gland increased significantly after the immobilization session in stressed groups during 30 and 50min as compared to control group. This increase probably is due to the emotional component of the immobilization stress. In this way, we suggested that the immobilization stress provoke increase in the biosynthesis of catecholamines in the adrenal gland from rats. However, the results shows that a maximum increase is reached at 30min of immobilization stress and then a decrement of catecholamines levels starts at 50min of the experimental design. This decline in catecholamines level may be consequence of adaptation to stress situations, an increase of the activity of the uptake systems and/or metabolization of catecholamines. In conclusion, these results suggest an effective participation of the adrenal glands to maintain the homeostasis of organism to the stressful conditions. © 2003 Elsevier Ltd. All rights reserved.

Immobilization of lipases and assay in continuous fixed bed reactor

Lipases are versatile enzymes regarding the range of reactions they catalyse and substrates on which they act. They are as well important as catalyst in organic synthesis. Their immobilization on appropriate supports confer them greater stability besides the possibility of operating in continuous reactors. In order to explore these abilities, the reactions involving hydrolysis of p-nitrophenyl acetate (PNPA) and transesterification of PNPA with n-butanol were chosen. Lipases from two different sources were assayed, namely: microbial (Candida rugosa, CRL, Sigma Type VII) and pancreatic (PPL, Sigma, Type 11). Two immobilization methods were also used, namely: 1) adsorption, using as support the following silica derivatives (150-300μm e 450μ): phenyl, epoxy, amino and without derivation, and 2) covalent binding, using glutaraldehyde as binding agent and silica amino as support. This later method led to better results. Hydrolytic activity was 6.1 U/gsupport for CRL and 0.97U/gsupport for PPL, and of transesterification, 2,8U/gsupport for CRL and 1,9U/gsupport for PPL. Stability of the immobilized enzyme as a function of temperature was evaluated for CRL at 40°C and 50°C and for PPL at 32°C and 40°C. The assays were initially carried out batchwise, both for soluble and immobilized enzymes, aiming to the obtention of parameters for the continues reactor. Lipases immobilized by covalent binding were used in the assays of operacional stability in continuos reactors. For PPL in aqueous medium, at 32°C, and CRL in organic medium at 40°C, both operating continuously, no significant loss of activity was detected along the analysis period of 17 days. In the case of CRL in aqueous medium at 40°C there was a loss of activity around 40% after 18 days. For PPL in organic medium at 40°C the loss was 33% after 20 days. Compairing both sources with each other, very different results were obtained. Higher activitiy was found for CRL, both for hydrolysis and for transesterification reactions, with higher stability in organic medium. PPL showed lower activity as well as higher stability in aqueous medium. The immobilization method by covalent binding showed to be the most appropriate. Immobilized lipases are therefore relatively stable both in aqueous and organic medium.

Early healing pattern of autogenous bone grafts with and without e-PTFE membranes: A histomorphometric study in rats

Objective. The aim of this study was to perform quantitative and qualitative analyses of the initial repair pattern of an autogenous bone block graft when covered or not with e-PTFE membranes. Study design. Sixty male Wistar rats received a bone graft plus an e-PTFE membrane (MB) or just the graft (B). A block graft was harvested from the animal's calvarium and was laid and stabilized on the external cortical area near the angle of the mandible. Descriptive histology and histomorphometric analyses were carried out and the data were analyzed statistically by ANOVA and the Tukey test, with the level of significance set at 5%. Results. The results for group B showed that there was bone loss during the healing period (B0 = 1.38, B45 = 1.05, F = 7.91 > F C = 3.02), that is, the initial volume of the graft decreased in time. Bone tissue loss was about 24%. In contrast, the MB group showed bone tissue gain along the observation period (MB0 = 1.54, MB45 = 2.40, F = 7.91 > FC = 3.02), meaning that the total volume of newly formed bone was greater than the original graft area. Bone tissue gain was approximately 55%. MB showed significantly greater bone gain when compared to B (B45 = 1.05, MB45 = 2.40, F = 39.86 > FC = 1.90). These significant differences between B and MB could already be observed after 21 days. Conclusions. The bone block graft underwent resorption at an early healing stage, while additional new bone formation was observed when the bone graft was covered with an e-PTFE membrane. © 2005 Mosby, Inc. All rights reserved.

Strategies for developing NADH detector based on meldola blue in different immobilization methods: A comparative study

A comparative study was accomplished between two immobilization methodologies for the meldola's blue (MB), used to decrease the oxidation potential of NADH. The study was performed with glassy carbon electrode by electropolymerization of pyrrole and MB through the technique of cyclic voltammetry being evaluated the effect of the number of cycles to define the thickness and the stability of the film. With 25 cycles it was obtained the best result. They were also investigated the modification of the graphite electrode with film formed by the system sol-gel PPO 300 or PPO 4000 for the dip-coating, through two methods: occlusion and adsorption. The best method was it of adsorption of MB during 10 min, following by deposit of the film PPO 300, favored for the time of drying (24 hours). The adsorption of MB followed by PPO 300 deposition presented broader linear range than PPy methodology. ©2006 Sociedade Brasileira de Química.

Utilization of neuromuscular electrical stimulation in the acute phase of ankle immobilization at 90°: Study in rats

Objective: To evaluate the skeletal muscle glycogen content and plasmatic concentration of interleukin -6 (IL-6), interleukin-4 (IL-4), interleukin-10 (IL-10) and tumor necrosis factor-alpha (TNF-α) in rats submitted to electrical stimulation sessions during the first three days of ankle immobilization at the position of 90°. Methods: Albinomale Wistar rats(3-4 months) were maintained in vivarium. conditions with food and water ad libitum, Submitted to 12 h photoperiodic cycles of light/dark, and distributed into 7 experimental groups (n = 6): control(C), immobilized 1 day(I1) immobilized 1 day and electrically stimulated(IE1) immobilized 2 days(12), immobilized 2 days and electrically stimulated(IE2), immobilized 3 days(13) and immobilized 3 days and electrically stimulated(IE3). Groups I utilized an acrylic resin orthesis model and groups electrically stimulated (IE) utilized the orthesis and a session of electrotherapy by a Dualpex 961 (biphasic quadratic pulse, 10 Hz, 0.4 ms, 5.0 mA, one 20 min session a day). After the experimental period, the rats were anesthetized with pentobarbital sodium(40 mg/kg) and a blood sample was colleted to evaluate the plasmatic concentration of interleukins by means of the radioimmunoassay method. The soleus and the white portion of the gastrocnemius muscle were colleted for glycogen reserves analysis(GLY). Other groups of rats were used to apply the glucose tolerance test(GTT) and insulin tolerance test(ITT). For statistical analysis, the Kolmogorov-Smirnov normality test followed by ANOVA and the Tukey tests were utilized, with a critical level established at 5%. Results: In ITT test, groups IE enhanced the skeletal muscle glucose uptake, but no changes were observed in GTT after the therapy session, which indicates that electrical stimulation is a sensibilizing method to augment skeletal muscle glucose uptake. The GLY reserves were reduced in I groups, which indicate that disuse altered insulin sensitivity and compromised energetic homeostasis. However. the IE groups displayed an augment in GLY content, suggesting that electrical stimulation restores the enzymatic pathways altered by immobilization. The improvement in GLY was accompanied by an elevation of the plasmatic concentration of IL-6 and TNF-α, showing the participation of these interleukins in the control of metabolic profile. Plasmatic concentrations of IL-10 were elevated only after 3 days of IE while IL-4 did not display any modifications. Conclusion: The results suggest that neuromuscular electricaf stimulation is an important toot in the maintenance of energetic, conditions of musculature submitted to immobilization, and presents multifactor mechanisms linked to interleukins action that converge to maintain the energetic equilibrium of the tissue in disuse.

Immunosensor based on immobilization of antigenic peptide NS5A-1 from HCV and silk fibroin in nanostructured films

The peptide NS5A-1 (PPLLESWKDPDYVPPWHG), derived from hepatitis C virus (HCV) NS5A protein, was immobilized into layer-by-layer (LbL) silk fibroin (SF) films. Deposition was monitored by UV-vis absorption measurements at each bilayer deposited. The interaction SF/peptide film induced secondary structure in NS5A-1 as indicated by fluorescence and circular dichroism (CD) measurements. Voltammetric sensor (SF/NS5A-1) properties were observed when the composite film was tested in the presence of anti-HCV. The peptide-silk fibroin interaction studied here showed new architectures for immunosensors based on antigenic peptides and SF as a suitable immobilization matrix. © 2013 American Chemical Society.

Amphiphilic planar membranes in ionic equilibrium: a study of pH position-dependent values

The pH values near a planar dissociating membrane are studied under a mean field approximation using the Poisson-Boltzmann equation and its linear form. The equations are solved in planar symmetry with the consideration that the charge density on the dissociating membrane surface results from an equilibrium process with the neighboring electrolyte. Results for the membrane dissociation degree are presented as a function of the electrolyte ionic strength and membrane surface charge density. Our calculations indicate that pH values have an appreciable variation within 2 nm from the membrane. It is shown that the dissociation process is enhanced due to the presence of bivalent ions and that pH values acquire better stability than in an electrolyte containing univalent ions.