Physalin pool from Physalis angulata L. leaves and physalin D inhibit P2X7 receptor function in vitro and acute lung injury in vivo.

P2X7 receptor promotes inflammatory response and neuropathic pain. New drugs capable of impairing inflammation and pain-reducing adverse effects extracted from plant extracts have been studied. Physalis angulate L. possesses traditional uses and exhibits antiparasitic, anti-inflammatory, antimicrobial, antinociceptive, antimalarial, antileishmanial, immunosuppressive, antiasthmatic. diuretic, and antitumor activities. The most representative phytochemical constituents identified with medicinal importance are the physalins and withanolides. However, the mechanism of anti-inflammatory action is scarce. Although some physalins and withanolides subtypes have anti-inflammatory activity, only four physalins subtypes (B, D, F, and G) have further studies. Therefore, we evaluated the crude ethanolic extract enriched with physalins B, D, F, and G from P. angulata leaves, a pool containing the physalins B, D, F, G, and the physalins individually, as P2X7 receptor antagonists. For this purpose, we evaluated ATP-induced dye uptake, macroscopic currents, and interleukin 1-ß (IL-1ß) in vitro. The crude extract and pool dose-dependently inhibited P2X7 receptor function. Thus, physalin B, D, F, and G individually evaluated for 5'-triphosphate (ATP)-induced dye uptake assay, whole-cell patch-clamp, and cytokine release showed distinct antagonist levels. Physalin D displayed higher potency and efficacy than physalin B, F, and G for all these parameters. In vivo mice model as ATP-induced paw edema was potently inhibited for physalin D, in contrast to physalin B, F, and G. ATP and lipopolysaccharide (LPS)-induced pleurisy in mice were reversed for physalin D treatment. Molecular modeling and computational simulation predicted the intermolecular interactions between the P2X7 receptor and physalin derivatives. In silico results indicated physalin D and F as a potent allosteric P2X7 receptor antagonist. These data confirm physalin D as a promisor source for developing a new P2X7 receptor antagonist with anti-inflammatory action.

Avaliação histomorfométrica do efeito do extrato aquoso de urucum (norbixina) no processo de cicatrização de feridas cutâneas em ratos / Histomorphometric evaluation of the effects of the aqueous extract of annatto (norbixin) on wound skin healing in rats

A norbixina é um carotenóide dicarboxílico hidrofílico presente no pericarpo das sementes do urucuzeiro (Bixa orellana L.). O urucum é comumente utilizado na indústria alimentícia e cosmética como corante natural. O objetivo deste trabalho foi avaliar o processo de reparo cicatricial de feridas cutâneas abertas de ratos tratadas com extrato aquoso de urucum contendo 2,5% de norbixina através de análise histomorfométrica. Para tanto, realizou-se feridas cutâneas no dorso de 32 ratos Wistar machos. Estas foram tratadas com extrato de urucum (grupo experimental, n = 16) e solução salina 0,9% (grupo controle, n = 16). Aos 3, 7, 14 e 21 dias após realização do procedimento cirúrgico, os animais foram sacrificados. Os preparados histológicos obtidos foram submetidos à técnica de coloração pela Hematoxilina-Eosina (H.E.) para contagem de células inflamatórias e de fibroblastos; corados pelo azul de toluidina 1% para contagem dos mastócitos e com picrossirius-red para avaliação das fibras colágenas totais. As imagens histológicas destas lâminas foram capturadas por câmera digital acoplada ao microscópio óptico, sob foco fixo e clareza de campo, obtendo-se 10 campos por lâmina com aumento final de 400X. As fotomicrografias foram avaliadas através do software ImageJ. Os resultados obtidos foram submetidos ao teste t de student sendo o valor de p considerado significativo para p< 0,05. O tratamento tópico com extrato de urucum utilizado aumentou o infiltrado inflamatório durante o 3º, 14º e 21º dia pós-operatório. O grupo tratado apresentou maior densidade vascular quando comparado ao controle a partir do 7º e menor quantidade de fibroblastos até o 14º dia pós-cirúrgico. Conclui-se que o extrato de urucum contendo 2,5% de norbixina não é inócuo aos tecidos cutâneos e possui efeitos pró-inflamatórios e pró-angiogênicos durante o processo de reparo tecidual cutâneo em ratos, interferindo no processo fisiológico de cicatrização.

Norbixin is a dicarboxylic water-soluble carotenoid present in the pericarp of the achiote (Bixa orellana L.) seed. Annatto is commonly used by the food and cosmetic industries as a natural pigment. The aim of this study was to evaluate the healing process of open dermal wounds of rats treated with an aqueous solution of annatto containing 2.5% of norbixin carried out by histomorphometric analysis. For this end, cutaneous wounds were made on the back of 32 male Wistar rats. The wounds were treated with annatto solution (experimental group, n = 16) and saline solution 0.9% (control group, n = 16). At 3, 7, 14 and 21 days after the surgical procedure, the animals were sacrificed. After histological preparation, the histological material was submitted to the staining technique with hematoxylin-eosin for the counting of inflammatory cells and fibroblasts, with toluidine blue 1% for mast cell counting and with picrosirius red for the total collagen fibers. The images of those histological slides were captured by a digital camera connected to an optical microscope, with fixed focal length and clear field; 10 fields were captured from each slide with final zoom of 400X. The photomicrographs were analyzed by the ImageJ software. The results were submitted to the Student's t-test and the value of p < 0.05 was considered as relevant. The topical treatment with the annatto solution increased the inflammatory infiltrate during the 3rd, 14th and 21st days after surgery. The experimental group presented increased vascular density compared to the control group after the 7th day and a smaller amount of fibroblasts up to the 14th day after surgery. We concluded that the annatto solution containing 2.5% of norbixin is not innocuous to skin tissues and has proinflammatory and proangiogenic effects during the process of skin wound healing in rats, interfering in the physiological healing process.

Design and field-scale implementation of an "on site" bioremediation treatment in PAH-polluted soil.

An "on site" bioremediation program was designed and implemented in soil polluted with polycyclic aromatic hydrocarbons (PAHs), especially naphthalene. We began by characterizing the soil's physical and chemical properties. A microbiological screening corroborated the presence of microorganisms capable of metabolizing PAHs. We then analyzed the viability of bioremediation by developing laboratory microcosms and pilot scale studies, to optimize the costs and time associated with remediation. The treatment assays were based on different types of biostimulants, such as a slow or fast-release fertilizer, combined with commercial surfactants. Once the feasibility of the biostimulation was confirmed, a real-scale bioremediation program was undertaken in 900 m(3) of contaminated soil. The three-step design reduced PAH contamination by 94.4% at the end of treatment (161 days). The decrease in pollutants was concomitant with the selection of autochthonous bacteria capable of degrading PAHs, with Bacillus and Pseudomonas the most abundant genera.

Short communication: In vitro assessment of erosive potential of energy drinks.

AIM: This in vitro study was to evaluate the endogenous pH, titratable acidity, total soluble solids content (TSSC) and nonreducing sugars of energy drinks. METHODS: Nine energy drinks (Bad Boy Power Drink, Red Bull, Red Bull Sugar Free, Flying Horse, Burn, Night Power, Flash Power, Flying Horse Light and 220V) were evaluated by a randomised experiment with 3 repetitions on each sample. pH analysis performed by potentiometry and buffering capacity was assessed by dilution of each drink. Increments of 0.1 N KOH were titrated until neutrality reached. TSSC readings were performed by Brix refractometry using an Abbé refractometer. RESULTS: pH values ranged from 1.52 (Flash Power) to 3.20 (Red Bull) and all drinks showed pH 5.5. Titratable acidity values ranged from 0.56 (220V) to 1.04 (Bad Boy Power Drink). Flying Horse Light presented the lowest TSSC content (1.66%) and Flying Horse presented the highest (12.58%). Non-reducing sugars values ranged from 0.00% (Red Bull Sugar Free and Flying Horse Light) to 54.33% (Flying Horse). CONCLUSION: The energy drinks evaluated have a high erosive potential, as they present low pH and a high non-reducing sugar content.

Protective effect of locally applied carvacrol gel on ligature-induced periodontitis in rats: a tapping mode AFM study.

The aims of this study were to test a locally applied carvacrol gel and determine its efficacy preventing alveolar bone loss in experimental periodontitis in rats by regular methodology to validate applicability the atomic force microscopy (AFM) as a novel morphology method on this model. Wistar rats were subjected to ligature around second, upper-left molars. Animals were treated carvacrol gel topically (CAG), immediately after Experimental Periodontitis Disease induction for 1' three-times/day for 11 days. A vehicle gel was utilized as control. The periodontium and the surrounding gingivae were examined at regular histopathology and by AFM method; the neutrophil influx into the gingivae was also assayed using myeloperoxidase activity. The bacterial flora was assessed through culture of the gingival tissue. Alveolar bone loss was significantly inhibited by CAG group compared to the Vehicle (V) group, the carvacrol gel treatment reduced tissue lesion at histopathology, with preservation of the periodontium, coupled to decreased myeloperoxidase activity in gingival tissue and also prevented the proliferation of periodontal microorganisms and the weight loss. The GAC treatment preserved alveolar bone resorption and showed anti-inflammatory and antibacterial activities in experimental periodontitis. Topographical changes in histological sections were seen bringing into high relief the periodontal structures, being a simple and cost-effective method for periodontal evaluation with ultrastructural resolution.

Thalamic vascular lesions. Risk factors and clinical course for infarcts and hemorrhages.

BACKGROUND AND PURPOSE: The data of patients with an MRI-confirmed diagnosis of thalamic stroke were analyzed to identify risk factors and to describe the clinical syndromes according to the thalamic arterial territory involved. METHODS: We examined the records of all patients with a diagnosis of thalamic stroke confirmed by MRI who attended the National Institute of Neurology and Neurosurgery "Manuel Velasco Suárez" from 1987 through 1995. A database containing demographic features, risk factors, clinical course, type and mechanism of stroke, and vascular territory was analyzed. RESULTS: Twenty-eight patients (19 men and 9 women; mean age, 48.2 years) had MRI-confirmed thalamic stroke. Infarct was present in 22 patients and hemorrhage in 6. Eleven (39.3%) patients were younger than 45 years. In these young patients, cigarette smoking was the main risk factor associated with the development of stroke. In young patients, isolated and multiple thalamic lesions were present in almost equal proportions. The remaining 17 patients were older adults (45 to 84 years of age). In such patients, most lesions were multiple and were associated with atherosclerosis-predisposing factors. The main distinguishing clinical feature for hemorrhages and paramedian infarcts in comparison with infarcts in the other thalamic territories was the presence of an altered level of consciousness. No patient died as a direct result of the stroke. CONCLUSIONS: Thalamic stroke commonly occurs in young adults. and its mechanism is often undetermined. Increased awareness of this type of stroke in young adults should lead to better delineation of its clinical features, course, and management.

Effect of Niclosamide (Bayluscide WP 70), Anacardium occidentale hexane extract and Euphorbia splendens latex on behavior of Biomphalaria glabrata (Say, 1818), under laboratory conditions.

The repellent effect of the molluscicides Niclosamide (Bayluscide WP 70), Anacardium occidentale and the latex of Euphorbia splendens on Biomphalaria glabrata was observed through the investigation of the occurrence of escape behavior among molluscs that were exposed to dosages lower than the LD 50. The total number of individuals out of water among the surviving snails in the control group provided a "Natural Escape Index". The comparison between this total and the total number of surviving snails in each group exposed to the different dosages of the molluscicides after 24 hr provided the "Molluscicide Escape Index" and the detection of a "Repellency Range" to these snails. The escape indexes for Niclosamide, A. occidentale and E. splendens were 10%, 6.22% and 6.44% respectively. Repellency occurred at the following concentration ranges: 0.01, 0.02 and 0.03 ppm Bayluscide, 0.1, 0.2 and 0.3 ppm A. occidentale and 0.05, 0.10, 0.15 and 0.20 ppm E. splendens. The Natural Escape Index obtained in the control group was zero.