NO production and potassium channels activation induced by Crotalus durissus cascavella underlie mesenteric artery relaxation.

Animal toxins are natural resources for pharmacological studies. The venom of Crotalus durissus cascavella (C.d. cascavella) may be a source in the bio-prospecting of new anti-hypertensive agents. The aim of this study was to investigate vascular effects of the venom of C.d. cascavella in normotensive rats. Studies were performed using isolated mesenteric artery segments and aortic endothelial cells. The cumulative administration of the venom of C.d. cascavella (0.001-30 µg/mL) on phenylephrine (Phe; 10 µM) pre-contracted rings induced a concentration-dependent vasorelaxation in the presence of vascular endothelium (Emax = 47.9 ± 5.0% n = 8), and its effect was almost abolished in the absence of endothelium (Emax = 5.8± 2.4% n = 5 (∗∗∗p < 0.001)). Tissue viability was maintained as there was no difference in the contractile capacity of rings before and after the administration of venom. The vasorelaxant effect of the venom was also abolished when arteries were pre-contracted with potassium chloride (KCl; 80 mM) (Emax = 6.4± 0.9% n = 5, ∗∗∗p < 0.001). When assessing the participation of endothelium-derived relaxing factors, it was noted that non-selective COX inhibition with indomethacin (10 µM) caused a significant reduction in the vasorelaxant effect of C.d. cascavella (*p < 0.05). When investigating the participation of NO released by endothelium, there was a significant reduction of the vasorelaxant effect of venom in rings treated with L-NAME (100 µM; Emax = 17.5± 2.2% n = 6; **p < 0.01). Similar results were noted in the presence of ODQ (10 µM), an inhibitor of soluble guanylyl cyclase (Emax = 11.2± 3.5%, n = 6) and PTIO (100 µM), a stable radical scavenger for nitric oxide (Emax = 10.77± 3.6%, n = 6). Moreover, the venom induced the release of NO by isolated aortic endothelial cells through amperometric studies. When assessing the participation of K+ channels on the vasodilatory response of the venom, tyrode solution with 20 mM of KCl caused a significant reduction in the relaxation response (p < 0.001) (Emax = 21.3 ± 8%, n = 7), as did inhibitor of delayed rectifier K+ channels (4-amynopiridine 1 mM; Emax = 9.5 ± 1.3, %, n = 5, ***p < 0.001), and vasorelaxation was almost abolished in the presence of Iberiotoxin (IbTx 100 nM). Therefore, these results suggest that the venom of C.d. cascavella induces vasorelaxation in superior mesenteric artery rings of normotensive rats in an endothelium-dependent manner. Specifically, the venom stimulates the generation of endothelium-derived relaxing factors, especially NO, and activates vascular smooth muscle hyperpolarization through K+ channels. These data illustrate that C.d. cascavella is a source of bioactive molecules and therefore has therapeutic potential in the treatment of cardiovascular diseases such as hypertension.

Simple sol-gel process to obtain silica-coated anatase particles with enhanced TiO2-SiO2 interfacial area.

In this study we prepared silica-titania composites with a low SiO2:TiO2 molar ratio. These materials were prepared using a simple sol-gel route in which a hydrothermal treatment was used to obtain mesoporous anatase particles. Pure titania was also synthetized for comparison purposes. These materials were examined by scanning and transmission electron microscopy (SEM and TEM, respectively), energy dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and nitrogen sorption tests. A thin silica coating was formed on the anatase particles. It was observed that the presence of this coating led to samples with an enhanced thermal stability. Indeed, the composites prepared in this work showed an anatase structure and a high specific surface area (SSA), even after their calcination at 800°C. Thus, we believe that the synthetized material present an outstanding SiO2-TiO2 interfacial area associated with a high amount of anatase particles which could improve its photoactive properties.

[Clinical, radiological and arthroscopic analysis of the ACL tear. A prospective study of 418 cases]. / Analyses clinique, radiologique et arthroscopique de la rupture du LCA. Etude prospective de 418 cas.

This prospective multicentric study concerns 418 anterior cruciate ligament tears. It correlates the arthroscopic data's and the clinical and radiological data's. Four types of anterior cruciate ligament tears were identified. Complete tears, postero lateral bundle preserved, healing on the posterior cruciate ligament and healing in the notch. The statistical correlations had shown a highest laxity in the complete tear group with a highest rate of soft Lachman and gross pivot shift, a highest incidence of medial meniscus tears was also noted and a longer delay between injury and surgery, 24 months for the complete tear group and seven months for the postero lateral bundle group. The mean medial compartment laxity, side to side, in the postero lateral bundle group was 4.93 mm and 7.93 mm in the complete tear group. These data could help the surgeon in his surgical planning especially in case of partial tears.

[Isokinetic assessment with two years follow-up of anterior cruciate ligament reconstruction with patellar tendon or hamstring tendons]. / Evaluation isocinétique à deux ans de ligamentoplasties du ligament croisé antérieur au tendon rotulien et aux ischiojambiers.

This retrospective multicentric study was designed to assess the outcome of quadriceps and hamstrings muscles two years after Anterior Cruciate Ligament (ACL) reconstruction and compare muscles recovery depending on the type of graft and individual variables like age, gender, level of sport, but also in terms of discomfort, pain and functional score. The results focused on the subjective and objective IKDC scores, SF36, the existence or not of subjective disorders and their location. The review included isokinetic muscle tests concentric and eccentric extensors/flexors but also internal rotators/external rotators with analysis of mean work and mean power. One hundred and twenty-seven patients were included with an average age 29 years (+/-10). They all had an ACL reconstruction with patellar tendon or hamstring tendon with single or double bundles. In the serie, the average muscles deficit at two years was 10% for the flexors and extensors but with a significant dispersion. Significant differences were not noted in the mean values of all parameters in term of sex or age (over 30 years or not), neither the type of sport, nor of clinical assessment (Class A and B of objective IKDC score), nor the existence of anterior knee pain. There was a relationship between the level of extensor or flexor recovery and the quality of functional results with minimal muscle deficits close to 5% if the IKDC score was over 90 and deficits falling to 15% in the group with IKDC score less than 90. The type of reconstruction (patellar tendon versus hamstrings) had an influence on the muscle deficit. For extensors, the recovery was the same in the two groups, more than 90% at two years and the distribution of these two populations by level of deficit was quite the same. For flexors, residual deficits were significantly higher in the hamstrings group on the three studied parameters whatever the speed and the type of contraction (concentric or eccentric) with an average deficit of 14 to 18%, while, in the patellar tendon group, there was a dominance over the opposite side of 2 to 3% in concentric contraction. The hamstrings deficit appears to be "harvest dependent". For internal rotators, a significantly higher deficit is observed in eccentric contraction for the hamstrings group. The residual hamstrings deficits were related to the number of tendons harvested: -7% when there was no harvest, 7% with one tendon harvested and 17% with two tendons harvested. The relationship between the level of recovery of the quadriceps muscle and hamstrings at two years and the quality of functional results incite, regarding the significantly higher deficit of flexors in ACL reconstructions with hamstrings, to change the rehabilitation programs and especially on early rehabilitation of hamstrings in eccentric mode in the early weeks postoperative considering the harvest site as an equivalent of muscle tear.

Design of novel hybrid organic-inorganic nanostructured biomaterials for immunoassay applications.

The purpose of this study was to develop novel hybrid organic-inorganic materials based on poly(vinyl alcohol) (PVA) polymer chemically crosslinked network to be tested as solid support on bovine herpesvirus immunoassay. Hybrids were synthesized by reacting PVA with three different alkoxysilanes modifying chemical groups: tetraethoxysilane (TEOS), 3-mercaptopropyltrimethoxysilane (MPTMS) and 3-glycidoxypropyltrimethoxysilane (GPTMS). PVA-derived hybrids were also modified by chemically crosslinking with glutaraldehyde (GA) during the synthesis reaction. In order to investigate the structure in the nanometer-scale, PVA-derived hybrids were characterized by using small-angle x-ray scattering synchrotron radiation (SAXS) and x-ray diffraction (XRD). PVA hybrids' chemical functionalities and their interaction with herpesviruses were also characterized by Fourier transform infrared spectroscopy (FTIR). The bioactivity assays were tested through enzyme linked immunosorbent assay (ELISA). SAXS results have indicated nano-ordered disperse domains for PVA hybrids with different x-ray scattering patterns for PVA polymer and PVA-derived hybrids. FTIR spectra have shown major vibration bands associated with organic-inorganic chemical groups present in the PVA, PVA-derived by silane modifier and PVA chemically crosslinked by GA. The immunoassay results have shown that PVA hybrids with chemically functionalized structures regulated to some extent the specific bioimmobilization of herpesvirus onto solid phase. We think that it is due to the overall balance of forces associated with van der Waals interaction, hydrophilic and hydrophobic forces and steric hindrance acting at the surface. PVA and PVA-derived hybrid materials were successfully produced with GA crosslinking in a nanometer-scale network. Also, such a PVA-based material could be advantageously used in immunoassays with enhanced specificity for diagnosis.

Biomaterial with chemically engineered surface for protein immobilization.

The last 3 decades have been a revolution in the area of sol-gel-derived materials. They can be used to encapsulate biomolecules such as enzymes, antibodies, hormones, and proteins retaining their functional state. Proteins can be immobilized in many ways but it is crucial that they retain their native conformational structure and, therefore, bioactivity. Porous silica gel matrixes with modified surfaces offer unlimited possibilities to control the protein-solid interaction behavior. The bioimmobilization process on sol-gel biomaterials with chemically engineered surface has driven applications on solid-phase materials, affinity chromatography, biosensors and many others. In the present work, we have aimed to produce surface-modified silica glass materials obtained via sol-gel route to be used as solid support on drug delivery systems and as solid-phase in immunodiagnostic. The functionalization process was carried out by reacting alkoxysilanes with 5 different silane surface modifying chemical groups: tetraethoxysilane (TEOS), 3-mercaptopropyltrimethoxysilane (MPTMS) and 3-aminopropyltriethoxysilane (APTES), 3-glycidoxypropyltrimethoxysilane (GPTMS) and 3-isocyanatopropyltriethoxysilane (ICPES). The bioactivity assays were based on two main tests: (a) An in vivo bioresponse of rats with sol-gel disk implants with insulin protein incorporated. In vivo tests with adult male rats were used to verify the immobilized insulin bioactivity after implantation of different biomaterial with functionalized surfaces. All surface modified materials have presented hypoglycemic peak response associated with the insulin bioactivity. (b) The produced solid-phase sol-gel disks with protein substrates were tested through Enzyme Linked Immuno Sorbent Assay (ELISA). The immunoassay results have showed that glasses with chemically functionalized surfaces regulated the extent of bioimmobilization of protein. The amine, thiol and hydroxyl terminated porous gels have showed significant interaction with the antibody-antigen, during the coupling process. We believe that it is due to balance of forces associated with Van der Waals interaction, hydrophilic and hydrophobic forces and steric hindrance acting at the surface. Therefore, such novel biomaterial could be advantageously used in drug delivery systems and in immunoassays of diagnostic kits.

In vitro release kinetics of proteins from bioactive foams.

This study describes an approach to obtaining 3-D scaffolds for tissue engineering that allows the incorporation and release of biologically active proteins to stimulate cell function. Laminin was adsorbed on the textured surfaces of binary 70S30C (70 mol % SiO(2), 30 mol % CaO) and ternary 58S (60 mol % SiO(2), 36 mol % CaO, 4 mol % P(2)O(5)) foams. The covalent bonds between the binding sites of the proteins and the ligands on the scaffolds' surfaces did not denaturate the proteins. In vitro studies show that the foams modified with chemical groups and coated with laminin were bioactive, as demonstrated by the formation of a crystalline hydroxy carbonate apatite (HCA) layer formed on the surfaces of the foams upon exposure to simulated body fluid (SBF). The release of proteins from the foams also was investigated. Sustained and controlled release from the scaffolds over a 30-day period was achieved. Laminin release from the bioactive foams followed the dissolution rate of the material network. These results suggest that bioactive foams have the potential to act as scaffolds for soft-tissue engineering with a controlled release of proteins that can induce tissue formation or regeneration.

Surface-modified 3D scaffolds for tissue engineering.

The aim of this work was to use sol-gel processing to develop bioactive materials to serve as scaffolds for tissue engineering that will allow the incorporation and release of proteins to stimulate cell function and tissue growth. We obtained organofunctionalized silica with large content of amine and mercaptan groups (up to 25%). The developed method can allow the incorporation and delivery of proteins at a controlled rate. We also produced bioactive foams with binary SiO(2)-CaO and ternary SiO(2)-CaO-P(2)O(5) compositions. In order to enhance peptide-material surface properties, the bioactive foams were modified with amine and mercaptan groups. These materials exhibit a highly interconnected macroporous network and high surface area. These textural features together with the incorporation of organic functionally groups may enable them to be used as scaffolds for the engineering of soft tissue.

Surface functionalization of porous glass networks: effects on bovine serum albumin and porcine insulin immobilization.

Biomolecules can be immobolized in many different ways. They can also be entrapped or tightly adsorbed within porous gels, clays, membranes, resins, and several other materials, but it is crucial that they retain their active conformation after the incorporation procedure. Porous gel matrixes with functionalized surfaces offer unlimited possibilities to control the protein-substrate interaction behavior. In the present work, we have studied the adsorption and the relative stability of bovine serum albumin (BSA) and porcine insulin(PI) incorporated in gels of SiO2 glass matrixes. The porous gel matrixes were obtained using tetramethoxysilane (TMOS)/metanol and functionalized with (3-mercaptopropyl) trimethoxysilane and (3-aminopropyl) triethoxysilane. The relative adsorption kinetics and stability of BSA and PI incorporated in glass networks were evaluated by immersion in phosphate buffer saline (PBS) and alkaline elution media for different periods of time. The kinetics of protein release from the gel matrix was monitored by UV-visible spectroscopy. A significantly larger PI release was observed compared to BSA in PBS solutions. We believe this is mainly associated with the difference on protein interactions with the modified surface, according to the characterization results of porosity, surface area, and contact angle of different functionalized gel matrixes. We could not observe any evidence of denaturation with either proteins after their desorption from gel matrixes using the ultraviolet spectroscopy technique. These results have also been confirmed with the strong bioactivity response from "in vivo" test conducted in rats, where porous gels with PI incorporated were implanted, showing that released proteins retained their native conformation.

[Dermatozooiasis in a large tropical country (nomenclature--practical approach) dermatozoiases (dermatozooparasitosis)]. / Dermatozoíases em um imenso país tropical (conduta prática-nomenclatura) Dermatozoíases (dermatozooparasitoses).

The author presents an extensive study of the "Dermatozooparasitoses" derived from his own personal experience and intends in this paper, to give practical directions in its therapeutics. Moreoever, he attempts to establish a nomenclature in a tropical and continental country like Brazil.