Polyvinylidene fluoride - Hydroxyapatite 0-3 biocomposite filaments processed by twin-screw extrusion.

Polyvinylidene fluoride - hydroxyapatite composite filaments were processed by twin-screw extrusion at different processing angular velocities and characterized by scanning electron and atomic force microscopies, differential scanning calorimetry and tensile tests. Polymer-ceramic composites with a 0-3 connectivity were successfully obtained. Regardless of the used processing parameters, all composite filaments present very similar melting (∼152°C) and solidification (∼139°C) points and elastic moduli (∼1.0 GPa) for hydroxyapatite as dispersed phase in the composite with concentrations up to 25 wt%, indicating that they are adequate for twin-screw extrusion and 3D printing. However, the yield strength (∼29 MPa), ultimate tensile strength (∼36 MPa) and tensile point (∼29 MPa) parameters are similar only for hydroxyapatite concentrations up to 15 wt%, once higher concentrations of hydroxyapatite as dispersed phase result in fragile samples (∼50% lower for each studied property).

Insights into the antiviral activity of phospholipases A2 (PLA2s) from snake venoms.

Viruses are associated with several human diseases that infect a large number of individuals, hence directly affecting global health and economy. Owing to the lack of efficient vaccines, antiviral therapy and emerging resistance strains, many viruses are considered as a potential threat to public health. Therefore, researches have been developed to identify new drug candidates for future treatments. Among them, antiviral research based on natural molecules is a promising approach. Phospholipases A2 (PLA2s) isolated from snake venom have shown significant antiviral activity against some viruses such as Dengue virus, Human Immunodeficiency virus, Hepatitis C virus and Yellow fever virus, and have emerged as an attractive alternative strategy for the development of novel antiviral therapy. Thus, this review provides an overview of remarkable findings involving PLA2s from snake venom that possess antiviral activity, and discusses the mechanisms of action mediated by PLA2s against different stages of virus replication cycle. Additionally, molecular docking simulations were performed by interacting between phospholipids from Dengue virus envelope and PLA2s from Bothrops asper snake venom. Studies on snake venom PLA2s highlight the potential use of these proteins for the development of broad-spectrum antiviral drugs.

Leishmania (Viannia) braziliensis in dogs in Brazil: epidemiology, co-infection, and clinical aspects.

Leishmaniasis is an endemic disease present in 98 countries. In Brazil, the northeast region accounts for approximately half of the cases in humans, and has experienced an increased number of positive cases in dogs. In this study, we investigated the epidemiology of canine leishmaniasis in the city of Ilhéus, Bahia, using serological and molecular techniques and evaluated the possible environmental risk factors and associated clinical signs. Blood samples were collected from 560 dogs in urban and peri-urban areas in Ilhéus, northeastern Brazil. Genomic DNA was extracted from the selected animals and subjected to molecular analysis using Leishmania species-specific primers and diagnosis of Trypanosoma cruzi. A total of 54.72% of dogs were positive for Leishmania braziliensis, and animals positive for both Leishmania infantum and T. cruzi were not identified. Hematologic variables were not statistically associated with cases of L. braziliensis. However, the positive animal group showed lower red blood cell and platelet counts and higher levels of urea and serum creatinine. Few dogs presented clinical signs compatible with the presence of Leishmania. Age of more than 2 years and specific hair colors were associated with positive results for L. braziliensis. The geoclimatic characteristics of the region may improve parasite survival, reproduction, and vectors. This may explain the higher rate of dogs identified as positive in this study.

Charge carriers and small-polaron migration as the origin of intrinsic dielectric anomalies in multiferroic TbMnO3 polycrystals.

Temperature-dependent and frequency-dependent dielectric investigations have been performed in TbMnO3 polycrystals sintered in either oxidative or reductive atmospheres. The results revealed the occurrence of two dielectric anomalies above 100 K, which are caused by the thermal activation of charge carriers and their motion in grain cores and grain boundaries. The temperature dependence of the bulk dc conductivity was also analysed and indicates that charge carriers move between inequivalent sites according to a variable-range-hopping mechanism. Also, a strong correlation between dielectric properties and crystalline structure was observed. Furthermore, a low-temperature dielectric relaxation, commonly reported in rare-earth manganite crystals, was observed in both samples. This relaxation follows the empirical Cole-Cole model and was attributed to small-polaron tunnelling. Polaron motion was observed to be affected by the magnetic transitions, structural properties and intrinsic anisotropies in TbMnO3. It is also worth mentioning that the dielectric anomaly due to motion of charge carriers in grain boundaries is the only one of extrinsic origin, while the anomalies related to carrier motion in grain cores and small-polaron tunnelling are intrinsic to TbMnO3.

Schinus terebinthifolius Raddi extract and linoleic acid from Passiflora edulis synergistically decrease melanin synthesis in B16 cells and reconstituted epidermis.

Several treatments for skin whitening are available today, but few of them are completely adequate, especially owing to the carcinogenic potential attributed to classical drugs like hydroquinone, arbutin and kojic acid. To provide an alternative and safer technology for whitening, we developed two botanical compounds originated from Brazilian biodiversity, an extract of Schinus terebinthifolius Raddi and a linoleic acid fraction isolated from Passiflora edulis oil. The whitening effect of these compounds was assessed using biochemical assays and in vitro models including cellular assays and equivalent skin. The results showed that S. terebinthifolius Raddi extract is able to reduce the tyrosinase activity in vitro, and the combination of this extract with linoleic acid is able to decrease the level of melanin produced by B16 cells cultured with melanocyte-stimulating hormone. Furthermore, melanin was also reduced in human reconstituted epidermis (containing melanocytes) treated with the compounds. The combination of the compounds may provide a synergistic positive whitening effect rather than their isolated use. Finally, we demonstrated that the performance of these mixed compounds is comparable to classical molecules used for skin whitening, as kojic acid. This new natural mixture could be considered an alternative therapeutic agent for treating hyperpigmentation and an effective component in whitening cosmetics.

Biochemical and biological evaluation of gyroxin isolated from Crotalus durissus terrificus venom

Gyroxin, a thrombin-like enzyme isolated from Crotalus durissus terrificus venom and capable of converting fibrinogen into fibrin, presents coagulant and neurotoxic activities. The aim of the present study was to evaluate such coagulant and toxic properties. Gyroxin was isolated using only two chromatographic steps - namely gel filtration (Sephadex G-75) and affinity (Benzamidine Sepharose 6B) - resulting in a sample of high purity, as evaluated by RP-HPLC C2/C18 and electrophoretic analysis that showed a molecular mass of 30 kDa. Gyroxin hydrolyzed specific chromogenic substrates, which caused it to be classified as a serine proteinase and thrombin-like enzyme. It was stable from pH 5.5 to 8.5 and inhibited by Mn²+, Cu²+, PMSF and benzamidine. Human plasma coagulation was more efficient at pH 6.0. An in vivo toxicity test showed that only behavioral alterations occurred, with no barrel rotation. Gyroxin was not able to block neuromuscular contraction in vitro, which suggests that its action, at the studied concentrations, has no effect on the peripheral nervous system.

A new fibrin sealant from Crotalus durissus terrificus venom: applications in medicine.

Fibrin sealant, a widely available tissue adhesive, has been used since 1940 in a variety of clinical applications. Commercially available fibrin sealant products are synthesized from bovine thrombin and human fibrinogen, which may transmit infectious diseases, and recipients may also develop antibodies against bovine thrombin. Bearing these disadvantages in mind, a new fibrin sealant was developed in 1989 by a group of researchers from the Center for the Study of Venoms and Venomous Animals, in Sao Paulo State, Brazil. The main purpose was to produce an adhesive fibrin without using human blood, to avoid transmitting infectious diseases. The components of this novel sealant were extracted from large animals and a serine proteinase extracted from Crotalus durissus terrificus snake venom. The applicability of this sealant was tested in animals and humans with beneficial results. The new fibrin sealant can be a useful tool clinically due to its flexibility and diversity of applications. This sealant is a biological and biodegradable product that (1) does not produce adverse reactions, (1) contains no human blood, (3) has a good adhesive capacity, (4) gives no transmission of infectious diseases, and (5) may be used as an adjuvant in conventional suture procedures. The effectiveness of this new fibrin sealant is reviewed and its development and employment are described.

Use of fibrin glue derived from snake venom in the repair of deep corneal ulcers: experimental study in dogs (Canis familiaris, Linnaeus, 1758)

Fibrin glue has been researched as an alternative method for tissue synthesis and is known for its capability to promote hemostasis at the application site, good approximation of wound edges and fast healing. The current study consisted in the application of fibrin glue derived from snake venom as treatment for experimental corneal ulcers. Twenty-one dogs had their corneas experimentally prepared through lamellar keratectomy (of standardized diameter and depth). Animals were divided into seven groups of three animals each. Six experimental groups were periodically evaluated and collection was carried out on the 1st, 3rd, 7th, 15th, 30th and 60th post-operative days, whereas one control group was evaluated throughout the experiment. Analyses consisted in the clinical evolution and in the histopathological study of all operated on eyes. Results indicated that fibrin glue was efficient in repairing keratectomy wounds in dogs and contributed to an earlier healing phenomenon, avoiding edema formation and keeping corneal clearness. The use of fibrin glue derived from snake venom showed to be easy to apply, feasible with animal models and of low cost, avoiding the lesion progress and allowing fast and appropriate corneal healing.

The effect of a remifentanil bolus on the bispectral index of the EEG (BIS) in anaesthetized patients independently from intubation and surgical stimuli.

BACKGROUND AND OBJECTIVE: Remifentanil boluses are used in different clinical situations and the effects on bispectral index monitoring are unclear. We analysed the effect of a remifentanil bolus on the bispectral index of the electroencephalogram (bispectral index) under total intravenous anaesthesia with propofol and remifentanil. METHODS: ASA I-III patients were included in this study. All patients received a 2 microg k g-1 remifentanil bolus in a period free from stimuli. Bispectral index and haemodynamic data were collected from an A-2000XP bispectral index monitor (every second) and an AS/3 Datex monitor (every 5 s). Bispectral index data were analysed using the area under the curve. Mean arterial pressure and heart rate were averaged at each 30-s period and analysed using analysis of variance. RESULTS: A total of 240 bispectral index values were obtained per patient. The area under the curve between 90 and 120 s after the bolus was significantly lower than the basal area under the curve (average of all areas before the bolus, P < 0.05). Mean arterial pressure and heart rate were significantly reduced from 96.4 +/- 19.9 mmHg at the time of the bolus to 74.2 +/- 16.6 mmHg 120 s after, and from 70 +/- 16.4 bpm at the time of the bolus to 61 +/- 13.6 bpm after (P < 0.001), respectively. CONCLUSIONS: There was a significant reduction in the areas under the curve between 90-120 s following the bolus. Heart rate and blood pressure also showed significant reductions. Thus, remifentanil bolus given under total intravenous anaesthesia with propofol and remifentanil decreases bispectral index, an effect independent of intubation and surgical stimuli.

[Hydroxyethyl starch 130/0.4 can be useful to prevent a hemodynamic response to a single dose of remifentanil]. / E1 hidroxietilalmidón (HES) 130/0.4 puede ser úitil en la prevención de la respuesta hemodinámica a un bolus de remifentanilo.

Intravenous propofol and remifentanil are often used in anesthesia. The combined use of these drugs tends to cause hemodynamic depression. We describe the absence of hemodynamic effects in response to infusion of propofol and remifentanil when hydroxyethyl starch (HES) 130/0.4 was also administered. During induction, because blood volume needed to be replaced, two patients aged 62 and 65 years received intravenous HES 130/0.4. They then received a single dose of 2 microg x kg(-1) of remifentanil during total intravenous anesthesia (TIVA) with propofol and remifentanil before placement of a Mayfield head holder. No changes in mean blood pressure or heart rate were observed in either patient after the remifentanil bolus when they have received HES 130/0.4 during TIVA with propofol and remifentanil HES 130/0.4 may play an active role in preventing a hemodynamic response to remifentanil bolus. This hypothesis should be tested in a randomized controlled trial.

Measurement of IL-10 serum levels in balb/c mice treated with beta-1, 3 polyglucose or sulfadiazine and acutely infected by Toxoplasma gondii

Acute infection by Toxoplasma gondii leads to suppression of cell-mediated immunity, facilitating chronic infection. One of the causes of immunosuppression is Interleukin-10 (IL-10) production. Glucan is used to stimulate phagocytosis. Our objective was to study IL-10 induction in male BALB/c mice with acute T. gondii BTU-2 strain infection, glucan immunostimulation, and sulfadiazine treatment. Animals were distributed into 7 groups: G1: infected with T. gondii; G2: infected with T. gondii and treated with sulfadiazine; G3: infected with T. gondii and immunostimulated with glucan; G4: infected with T. gondii, immunostimulated with glucan, and treated with sulfadiazine; G5: imunostimulated with glucan; G6: treated with saline; and G7: treated with sulfadiazine. IL-10 levels were determined by ELISA; the highest levels were found in G2, G3 and G4, and the lowest in G1 (p<0.001). Groups G1 to G5 and G7 had substantially higher levels than G6 (p<0.001). In this study, the highest IL-10 levels were found in groups treated with glucan

Heparan sulfate and control of cell division: adhesion and proliferation of mutant CHO-745 cells lacking xylosyl transferase

We have examined the role of cell surface glycosaminoglycans in cell division: adhesion and proliferation of Chinese hamster ovary (CHO) cells. We used both wild-type (CHO-K1) cells and a mutant (CHO-745) which is deficient in the synthesis of proteoglycans due to lack of activity of xylosyl transferase. Using different amounts of wild-type and mutant cells, little adhesion was observed in the presence of laminin and type I collagen. However, when fibronectin or vitronectin was used as substrate, there was an enhancement in the adhesion of wild-type and mutant cells. Only CHO-K1 cells showed a time-dependent adhesion on type IV collagen. These results suggest that the two cell lines present different adhesive profiles. Several lines of experimental evidence suggest that heparan sulfate proteoglycans play a role in cell adhesion as positive modulators of cell proliferation and as key participants in the process of cell division. Proliferation and cell cycle assays clearly demonstrate that a decrease in the amount of glycosaminoglycans does not inhibit the proliferation of mutant CHO-745 cells when compared to the wild type CHO-K1, in agreement with the findings that both CHO-K1 and CHO-745 cells take 8 h to enter the S phase

Heparan sulfate and control of cell division: adhesion and proliferation of mutant CHO-745 cells lacking xylosyl transferase.

We have examined the role of cell surface glycosaminoglycans in cell division: adhesion and proliferation of Chinese hamster ovary (CHO) cells. We used both wild-type (CHO-K1) cells and a mutant (CHO-745) which is deficient in the synthesis of proteoglycans due to lack of activity of xylosyl transferase. Using different amounts of wild-type and mutant cells, little adhesion was observed in the presence of laminin and type I collagen. However, when fibronectin or vitronectin was used as substrate, there was an enhancement in the adhesion of wild-type and mutant cells. Only CHO-K1 cells showed a time-dependent adhesion on type IV collagen. These results suggest that the two cell lines present different adhesive profiles. Several lines of experimental evidence suggest that heparan sulfate proteoglycans play a role in cell adhesion as positive modulators of cell proliferation and as key participants in the process of cell division. Proliferation and cell cycle assays clearly demonstrate that a decrease in the amount of glycosaminoglycans does not inhibit the proliferation of mutant CHO-745 cells when compared to the wild type CHO-K1, in agreement with the findings that both CHO-K1 and CHO-745 cells take 8 h to enter the S phase.

Surgical adhesives

The authors have performed a literature review of surgical adhesives, such as cyanoacrylate, collagen gelatin, and fibrin glue. They have included different types of commercial and non-commercial fibrin sealants and have reported on the different components in these adhesives, such as fibrinogen, cryoprecipitate, bovine thrombin, and thrombin-like fraction of snake venom.

An evaluation by rat colon anastomosis of the efficacy of fibrin glue derived from snake venom

The objective of this study was to evaluate the efficacy of fibrin adhesive made up of snake venom and bubaline fibrinogen by rat colon anastomosis. Eighty rats were randomly assigned into 2 experimental groups: GI control (anastomosis with extramucous interrupted suture) and GII (repair suture + fibrin glue). The animals were studied at the following 4 times: T0 - preoperative - T1 - 7th day postoperative, T2 - 14th day postoperative, and T3 - 21th day postoperative. The macroscopic characteristics of the intestinal segment open and closed anastomosis and the bursting strength of the anastomosed segments were observed at each of the above times. The results showed that the anastomosed segments coapted and there was no difference in the bursting strength values between the 2 groups. There was a decrease in the bursting strength values up until de 7th day postoperative in both groups with its progressive increase at the other times. Although important experimental studies using large animals are needed for a better evaluation of tissue repair processes, this adhesive may become a valuable tool for use in anastomosis.

The use of snake venom derived fibrin glue in hysterorrhaphy of ovine caesarean surgery

Fibrin glue has been used on its own or in conjunction with suturing materials to promote hemostasis, reduce adherence, strengthen the wound site, and improve healing. Snake venom derived fibrin glue was evaluated as an alternative to conventional uterine suturing after ovine caesarean surgey. Twenty-eight pregnant ewes of known mating date were used. The animals submitted to conventional caesaream sections showed a better wound healing process. As expected, all the operated animals had retained placenta, compromissing coaptation of wound edges. This had a strong influence in the results observed with the animals in which fibrin glue was used. The animals were divided into four groups GI, GII, GIII, and GIV and sacrificed, respectively, 3, 7, 15, and 30 days after surgery for macro and microscopic examination of the uterus. From each group, six animals underwent surgery using fibrin glue and four animals were submitted to comparative conventional hysterorrhaphy using catgut.

The evaluation of clotting time in bovine thrombin, reptilase©, and thrombin-like fraction of Crotalus durissus terrificus venom using bovine, equine, ovine, bubaline and human cryoprecipitates

The objective of this study was to evaluate the effects of the thrombin-like fraction of Crotalus durissus terrificus venom, Reptilase©, and bovine thrombin of fibrinogen polls on bovine, equine, ovine, bubaline and human cryoprecipitates. The authors also made a comparative study between animal and human cryoprecipitates to see if there was any possibility of future use in medicine. Fibrinogen levels in cryoprecipitate were studied using 48 blood samples obtained as follows: 12 samples from humans, 9 from bovine, 10 from equine, 10 from ovine and 7 from bubaline. The results obtained showed average levels of 375.50 mg per cent for humans, 218.33 mg per cent for bovine, 240.80 mg per cent for equine, 267.70 mg per cent for ovine and 664.00 mg per cent for bubaline. Upon the formation of pools of human and animals fibrinogens, the following results were obtained: 435 mg per cent for humans, 444 mg per cent for bovine, 337 mg per cent por equine, 390 mg per cent for ovine and 530 mg per cent for bubaline. Statistical analysis (using the analysis of variance for entirely randomized experiment for the calculation of F statistics) demonstrated that the bubaline fibrinogen level was higher than that of human, and both were higher than those of ovine, equine, and bovine. Clotting times were determined using different dilutions of bovine thrombin, thrombin-like fraction of Crotalus durissus terrificus venom, and Reptilase©. Comparing these clotting times, results for human and bovine were found to be very similar, whereas using equine, ovine and bubaline the results above a dilution of 1:3 were markedly different. The results obtained permitted the following conclusions to be drawn show that: 1) bovine thrombin presented better interactivity with fibrinogen extracted both from human and bovine cryoprecipitates; 2) there was similar behavior when bovine thrombin was substituted for Reptilase© and for the thrombin-like fraction of Crotalus durissus terrificus venom; 3) cryoprecipitate from bovine can, in special circumstances, substitute human cryoprecipitate in medical practice; 4) human and bovine cryoprecipitates can be used with both Reptilase© and Crotalus durissus terrificus fractions using a dilution up to 1:5; 5) the use of bovine cryoprecipitate can be recomended using either bovine thrombin, Reptilase©, or thrombin-like fraction of Crotalus durissus terrificus venom.

Minimum fragments of the heparin molecule able to produce the accumulation and change of the sulfation pattern of an antithrombotic heparan sulfate from endothelial cells.

Heparin and low molecular weight heparins stimulate two to three fold the accumulation of an antithrombotic heparan sulfate secreted by endothelial cells in culture. This led us to search for the minimum structural requirements of the heparin molecule able to elicit the enhancement of the heparan sulfate. Fragments were prepared from heparin by degradation with bacterial heparinase and heparitinases. A heparin pentasulfated tetrasaccharide was shown to be the minimum structural sequence able to enhance two to three fold the secretion of heparan sulfate by endothelial cells. The stimulation is specific for the endothelial cell, is concentration dependent and the effect is already noticed after one hour of exposure of the cells to heparin and the tetrasaccharide. Degradation of the [35S]-heparan sulfate synthesized in the presence of heparin or the tetrasaccharide has shown a higher degree of sulfation of its iduronic acid residues.