Snake venom phospholipase A(2) inhibitors: Medicinal chemistry and therapeutic potential

Phospholipases A(2) (PLA(2)s) are commonly found in snake venoms from Viperidae, Hydrophidae and Elaphidae families and have been extensively studied due to their pharmacological and physiopathological effects in living organisms. This article reports a review on natural and artificial inhibitors of enzymatic, toxic and pharmacological effects induced by snake venom PLA(2)s. These inhibitors act on PLA(2)S through different mechanisms, most of them still not completely understood, including binding to specific domains, denaturation, modification of specific amino acid residues and others. Several substances have been evaluated regarding their effects against snake venoms and isolated toxins, including plant extracts and compounds from marine animals, mammals and snakes serum plasma, in addition to poly or monoclonal antibodies and several synthetic molecules. Research involving these inhibitors may be useful to understand the mechanism of action of PLA(2)s and their role in envenomations caused by snake bite. Furthermore, the biotechnological potential of PLA(2) inhibitors may provide therapeutic molecular models with antiophidian activity to supplement the conventional serum therapy against these multifunctional enzymes.

Inteins in pathogenic fungi: a phylogenetic tool and perspectives for therapeutic applications

Inteins or internal proteins are coding sequences that are transcribed and translated with flanking sequences (exteins). After translation, the inteins are excised by an autocatalytic process and the host protein assumes its normal conformation and develops its expected function. These parasitic genetic elements have been found in important, conserved proteins in all three domains of life. Most of the eukaryotic inteins are present in the fungi kingdom and the PRP8 intein is one of the most widespread inteins, occurring in important pathogens such as Cryptococcus neoformans (varieties grubii and neoformans), Cryptococcus gattii, Histoplasma capsulatum and Paracoccidioides brasiliensis. The knowledge of conserved and non-conserved domains in inteins have opened up new opportunities for the study of population variability in pathogenic fungi, including their phylogenetic relationships and recognition or diagnoses of species. Furthermore, inteins in pathogenic fungi should also be considered a promising therapeutic drug target, since once the autocatalytic splicing is inhibited, the host protein, which is typically vital, will not be able to perform its normal function and the fungal cell will not survive or reproduce.

Kinetics of venom and antivenom serum levels, clinical evaluation and therapeutic effectiveness in dogs inoculated with Crotalus durissus terrificus venom

This work evaluated the clinical and therapeutic aspects as well as serum levels of venom and antivenom IgG by enzyme-linked immunosorbent assay (ELISA) in experimental envenomation of dogs with Crotalus durissus terrificus venom. Twenty-eight mixed breed adult dogs were divided into four groups of seven animals each, Group I: only venom; Group II, venom + 50 ml of anti-bothropic-crotalic serum (50mg) + fluid therapy; Group III, venom + 50 ml of anti-bothropic-crotalic serum + fluid therapy + urine alkalination; Group IV, 50 ml of anti-bothropic-crotalic serum. The lyophilized venom of Crotalus durissus terrificus was reconstituted in saline solution and subcutaneously inoculated at the dose of 1mg/kg body weight. The dogs presented clinical signs of local pain, weakness, mandibular ptosis, mydriasis, emesis and salivation. The venom levels detected by ELISA ranged from 0 to 90ng/ml, according to the severity of the clinical signs. Serum antivenom ranged from 0 to 3ug/ml and was detected for up to 138h after treatment. ELISA results showed the effectiveness of the serum therapy for the venom neutralization.

Chloroquine is therapeutic in murine experimental model of paracoccidioidomycosis

Chloroquine, due to its basic properties, has been shown to prevent the release of iron from holotransferrin, thereby interfering with normal iron metabolism in a variety of cell types. We have studied the effects of chloroquine on the evolution of experimental paracoccidioidomycosis by evaluating the viable fungal recovery from lung, liver and spleen from infected mice and H2O2, NO production, tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, IL-10 levels and transferrin receptor (TfR) expression from uninfected and infected peritoneal macrophages. Chloroquine caused a significant decrease in the viable fungal recovery from all organs tested, during all periods of evaluation. Peritoneal macrophages from chloroquine-treated infected mice showed higher H2O2 production and TfR expression, and decreased levels of NO, endogenous and stimulated-TNF-alpha, IL-6 and IL-10 during the three evaluated periods. However, despite its suppressor effects on the macrophage function, the chloroquine therapeutic effect upon murine paracoccidioidomycosis was probably due to its effect on iron metabolism, blocking iron uptake by cells, and consequently restricting iron to fungus growth and survival.

Therapeutic and pathogenetic animal models for Dolichos pruriens

The therapeutic and pathogenetic effects of Dolichos pruriens were evaluated using experimental models in rats. In the therapeutic experiment Wistar rats were housed in a heated environment (25 +/- 3 degrees C) to induce itch, and treated with ascending potencies D. pruriens (6 cH, 9 cH, 12 cH and 30 cH), each for 10 days. The positive control group received vehicle (ethanol 30% in water). The negative control group received no treatment and were kept at a standard temperature.In the pathogenetic experiment, all animals were kept at a temperature of 20+/-3 degrees C and treated for 30 consecutive days with D. pruriens 6 or 30 cH, or ethanol vehicle, or no treatment. The experiments were performed blind.The statistical analysis used Bartlett's test, followed by ANOVA/Tuckey-Krammer or Kruskal-Wallis/Dunn. The results point to the existence of therapeutic effects, with inhibition of the itching, skin lesions and fur thinning produced by heat, more evident in later observations, with the 9 12, and 30 cH potencies (Kruskal-Wallis/Dunn; P = 0.001). No changes were observed in the other parameters, such as open field activity and laterality of the itching. In the pathogenetic experiment, no changes were observed in any parameters examined. We conclude that the proposed experimental model demonstrates the therapeutic effect of D. pruriens, but not its pathogenetic effects.

Effect of continuous and pulsed therapeutic ultrasound in the appearance of local recurrence of mammary cancer in rats

Purpose: Ultrasound (US) therapy is an elect rot hermotherapeutic modality that uses US energy to provoke physical and chemical alterations. US therapy has been widely used in physical therapy. However in clinical practice, it is contra-indicated in cancer patients due to the possibility of exacerbating tumor growth.Methods: Sixty-eight female Sprague-Dawley rats bred in UNIFAE vivarium were studied. At 50 days of age, 7, 12-dimetylbenz(a)anthracene (7, 12-DMBA) was administered to 35 rats by gastric gavage to induce mammary cancer After 90 days the mammary glands of the rats belonging to the group with mammary cancer induction and stimulated by US were removed. Animals received either continuous or pulsed US. US waves were generated at a frequency of 1 MHz during 10 days, with an intensity dose of 0.5 W in the continuous group, and 0.9 W (duty cycle: 20%) in the pulsed group.Results: Among the rats treated with continuous US, 44.4% developed local recurrence, while among the rats treated with pulsed US, 22.2% had local tumor recurrence (p < 0.05). No evidence of distant metastases was shown in any of the rats studied.Conclusion: The use ofcontinuous and pulsed therapeutic US promoted the development of local recurrence of mammary cancer in female Sprague-Dawley rats in the postoperative period.

Design of novel iron compounds as potential therapeutic agents against tuberculosis

In the search for new therapeutic tools against tuberculosis two novel iron complexes, [Fe(L-H)3], with 3-aminoquinoxaline-2-carbonitrile N(1),N(4)-dioxide derivatives (L) as ligands, were synthesized, characterized by a combination of techniques, and in vitro evaluated. Results were compared with those previously reported for two analogous iron complexes of other ligands of the same family of quinoxaline derivatives. In addition, the complexes were studied by cyclic voltammetry and EPR spectroscopy. Cyclic voltammograms of the iron compounds showed several cathodic processes which were attributed to the reduction of the metal center (Fe(III)/Fe(II)) and the coordinated ligand. EPR signals were characteristic of magnetically isolated high-spin Fe(III) in a rhombic environment and arise from transitions between m(s) = +/- 1/2 (geff-9) or m(s) = +/- 3/2 (g(eff)similar to 4.3) states. Mossbauer experiments showed hyperfine parameters that are typical of high-spin Fe(III) ions in a not too distorted environment. The novel complexes showed in vitro growth inhibitory activity on Mycobacterium tuberculosis H(37)Rv (ATCC 27294), together with very low unspecific cytotoxicity on eukaryotic cells (cultured murine cell line J774). Both complexes showed higher inhibitory effects on M. tuberculosis than the "second-line" therapeutic drugs. (C) 2010 Elsevier B.V. All rights reserved.

Calibration of therapeutic ultrasound equipment

Temporal and spatial acoustic intensity (SATA) of therapeutic ultrasound (US) equipment should be monitored periodically. In order to evaluate the conditions of US equipment in use in the city of Piracicaba-Sao Paulo, Brazil, 31 machines - representing all Brazilian manufacturers - were analysed under continuous and pulsed conditions at a frequency of 1 MHz. Data about temporal and spatial acoustic intensity were collected and the use of equipment was surveyed. Intensities of 0.1, 0.2, 0.5, 0.8, 1.0, 1.5, 2.0, 2.5 and 3.0 Wcm -2, indicated on the equipment panel were analysed using a previously calibrated digital radiation pressure scale, model UPM-DT-1 (Ohmic Instruments Co). The acoustic intensity (I) results were expressed as superior and inferior quartile ranges for transducers with metal surfaces of 9 cm 2 and an effective radiation area (ERA) Of 4 cm 2. The results under continuous conditions were: I 0.1 = -20.0% and -96%. I 0.2 = -3.1% and -83.7%. I 0.5 = -35.0% and -86.5%. I 0.8 = -37.5% and -71.0%. I 2.5 = -49.0% and -69.5%. I 3.0 = -58.1% and -77.6%. For pulsed conditions, intensities were: I 0.1 = -40.0% and -86.2%. I 1.0 = -50.0% and -86.5%. I 1.5 = -62.5% and -82.5%. I 2.0 = -62.5% and -81.6%. I 2.5 = -64.7% and -88.8%. I 3.0 = -87.1% and -94.8%. In reply to the questionnaire drawn up to check the conditions of use of equipment, all users reported the use of hydrosoluble gel as a coupling medium and none had carried out previous calibrations. Most users used intensities in the range of 0.4. to 1.0 Wcm -2 and used machines for 300 to 400 minutes per week. The majority of machines had been bought during the previous seven years and weekly use ranged from less than 100 minutes to 700 minutes (11 hours 40 minutes). Findings confirm previous observations of discrepancy between the intensity indicated on the equipment panel and that emitted by the transducer and highlight the necessity for periodic evaluations of US equipment.

Therapeutic response of benzodiazepine, orphenadrine citrate and occlusal splint association in TMD pain

Loss of function, muscle inflammation, and pain are some of the signs and symptoms of temporomandibular dysfunction (TMD). Pharmacological strategies to minimize the clinical manifestation of these disorders often focus on blocking or inhibiting the pain-causing symptom. Resources such as muscle-relaxants, anxiety-relief drugs, and splint therapy are often used to reduce muscular hyperactivity related to TMD muscle pain. This study compares the effect of a randomly ordered association of occlusal splint therapy (S), nonsteroid anti-inflammatory with a muscle-relaxant drug (orphenadrine citrate) (O), and an anxiety-relief drug (benzodiazepine) (B), to ease painful TMD muscle symptoms. Clinical and anamnestic analyses were recorded in accordance with the Helkimo TMD index and applied before and after treatments. Twenty-one group two Helkimo TMD adult female patients were treated, all of whom were subjected to the three random therapeutic associations proposed: SBO, BOS, and OSB. The same operator applied the three specific associations over a period of 21 days in the proposed sequence, seven days for each therapy. The results show that all the groups presented the best results in terms of relief from pain after the therapeutic association (28.5% showed a decrease and 47.6% showed an absence of symptoms). No significant difference was observed among association therapeutic protocols. Copyright © 2003 by CHROMA, Inc.

Comparative therapeutic use of Risedronate and Calcarea phosphorica - Allopathy versus homeopathy - In bone repair in castrated rats

Osteoporosis, a disease characterized by progressive bone loss, has been the target of several studies in the past few years. It results in a much higher risk for fractures and might cause slower bone lesion healing. The aim of this work was to study the effects of Risedronate (allopathic medicine) and Calcarea phosphorica 6CH (homeopathic medicine) on the repair of bone lesions in male rats with osteoporosis induced by castration. Eighty-four three-month-old rats were used divided into four groups of twenty-one animals each. Three groups where castrated and one group was submitted to Sham surgery. One month later, cortical lesions were made in all animals' tibiae and, after one day, the different experimental treatments began according to the following groups: CR - castrated/Risedronate (1 mg/kg/day); CCp - castrated/ Calcarea phosphorica 6CH (3 drops/day); CP - castrated/placebo and SP - Sham/placebo. The animals were sacrificed at seven, fourteen and twenty-eight days after the beginning of the treatments and had their tibiae removed. Digital radiographs of the tibiae were taken and analyzed in order to evaluate the optical density of the defect area. Then, they were decalcified and processed for histological and histomorphometrical analysis. The data were submitted to ANOVA, and to the Tukey and Dunnett tests (5%). The allopathic and homeopathic treatments led to different bone formation as regards remodeling and maturation aspects. Further research is necessary to access the resistance and quality of the newly formed bone.