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Made available in DSpace on 2018-11-29T07:11:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-03-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Bone grafts are used in the medical-surgical field for anatomical and functional reconstruction of lost bone areas, aiding the bone repair process by osteogenesis, osteinduction and osteoconduction. New materials such as F1 (fraction 1) protein extracted from the rubber tree Hevea brasiliensis have been investigated and currently present important properties for tissue repair, and are associated with neoangiogenesis, promoting cell adhesion and extracellular matrix formation. The main objective of this study was to investigate the association of F1 protein to different bone grafts in the repair of critical bone defects in the calvaria of Wistar rats. A total of 112 Wistar rats were divided as follows: autograft (AuG), allograft (AlG), xenograft (XeG), autograft/F1 (AuG-F1), allograft/F1 (AlG-F1), xenograft/F1 (XeG-F1), F1(F1), control (CTL), with a waiting period of 4 and 6 weeks (w). The stereological AuG, AlG, AuG-F1 and AlG-F1 results had greater bone neoformation (p < 0.05). For immunohistochemistry, the angiogenic and osteogenic factors were higher for AuG-F1 and AlG-F1. TRAP positive cells were higher in XeG-F1 and AlG (37 +/- 9.53, 13.3 +/- 4.16) (4 w) and XeG, AlG-F1 and XeG-F1 (20.33 +/- 7.37; 15.25 +/- 6.02, 19.33 +/- 3.21) (6 w). For zymography, F1 showed increased gelatinolytic activity of MMP-2 and -9. It was concluded that the bone graft associated or not with F1 increases the angiogenic and osteogenic, biochemical and stereological factors. Univ Sao Paulo, Sch Med Ribeirao Preto, Ribeirao Preto, SP, Brazil Sao Paulo State Univ Julio Mesquita Filho, Aracatuba, SP, Brazil Univ Sao Paulo, Sch Dent Ribeirao Preto, Ribeirao Preto, SP, Brazil Sao Paulo State Univ Julio Mesquita Filho, Aracatuba, SP, Brazil FAPESP: 2012/03310-8 FAPESP: 2012/225321

Introduction: In this study we evaluated the characteristics of the tibialis anterior muscle after sciatic nerve crush and treatment with low-level laser therapy (LLLT) or the protein from natural latex (P1). Methods: We studied the following 6 groups of male Wistar rats: control (CG); exposed nerve (EG); injured nerve (IG); injured nerve with LLLT (LG); injured nerve with P1 (PG); and injured nerve with P1 and LLLT (LPG). Results: After 4 weeks, muscle morphology showed improvement in the treated groups; after 8 weeks, the treated groups resembled controls, especially the PG. Morphometry revealed muscle fiber atrophy after nerve injury, with time-dependent recovery. Histochemical analysis revealed increased intermediate fiber area. The PG was more similar to controls with NADH staining, whereas the LPG more closely resembled controls with SDH staining. Conclusion: Treatment using only P1 proved most efficient, revealing a negative interaction between P1 and LLLT. Muscle Nerve 52: 869–875, 2015

Excitatory amino acid transporters (EAATs) regulate glutamatergic signal transmission by clearing extracellular glutamate. Dysfunction of these transporters has been implicated in the pathogenesis of various neurological disorders. Previous studies have shown that venom from the spider Parawixia bistriata and a purified compound (Parawixin1) stimulate EAAT2 activity and protect retinal tissue from ischemic damage. In the present study, the EAAT2 subtype specificity of this compound was explored, employing chimeric proteins between EAAT2 and EAAT3 transporter subtypes and mutants to characterize the structural region targeted by the compound. This identified a critical residue (Histidine-71 in EAAT2 and Serine-45 in EAAT3) in transmembrane domain 2 (TM2) to be important for the selectivity between EAAT2 and EAAT3 and for the activity of the venom. Using the identified residue in TM2 as a structural anchor, several neighboring amino acids within TM5 and TM8 were identified to also be important for the activity of the venom. This structural domain of the transporter lies at the interface of the rigid trimerization domain and the central substrate-binding transport domain. Our studies suggest that the mechanism of glutamate transport enhancement involves an interaction with the transporter that facilitates the movement of the transport domain. We identified a domain (purple star) in the glutamate transporter EAAT2 that is important for transport stimulation through a spider venom, and suggest a mechanism for enhanced transporter function through facilitated substrate translocation (arrow). Because the dysfunction of glutamate transporters is implicated in the pathogenesis of neurological disorders, understanding the mechanisms of enhanced transport could have therapeutic implications.

RESUMO Objetivo: Estudar o uso da biomembrana de latex e o transplante conjuntival autologo na cicatrizacao conjuntival em coelhos. Metodos: Em nove coelhos albinos, neo-zelandeses, machos foram removidas areas retangulares identicas, do quadrante supero nasal, adjacente ao limbo, de ambos os olhos. As areas desnudas da camada esclerotica nos olhos direitos foram recobertas com biomembrana de latex e a dos olhos esquerdos com enxerto conjuntival autologo. Os animais foram sacrificados em grupos de tres, aos 7, 14 e 21 dias apos a cirurgia. Os tecidos do local cirurgico, incluindo a cornea, foram fixados em formaldeido, antes de serem processados em parafina e corados com hematoxilina e eosina. A natureza e a intensidade da resposta inflamatoria e o padrao de epitelizacao da superficie conjuntival foram avaliados sob microscopia optica, em secoes histologicas longitudinais, passando pelo centro dos especimes anatomicos. Resultados: Ate o decimo quarto dia pos-operatorio, o grupo que recebeu a biomembrana apresentou reacao inflamatoria mais intensa do que o grupo com auto enxerto conjuntival. Aos 14 dias, os olhos com biomembrana apresentavam-se menos inflamados e com estroma mais espesso do que aos 7 dias. Aos 21 dias, a reparacao conjuntival de ambos os grupos apresentavam caracteristicas semelhantes. Conclusao: Apesar de apresentar uma cicatrizacao mais lenta, a biomembrana de latex se mostrou tao eficaz quanto o auto enxerto conjuntival na reconstrucao da superficie ocular apos tres semanas de cicatrizacao pos-operatoria. Devido as suas baixas toxicidade e alergenicidade, este material parece ser uma opcao terapeutica promissora na reconstrucao da conjuntiva.ABSTRACT Purpose: To study a latex biomembrane and conjunctival autograft with regard to the promotion of conjunctival healing in rabbits. Methods The study included nine male albino rabbits. In these rabbits, a rectangular area of the conjunctiva was surgically removed from the superonasal quadrant adjacent to the limbus in both eyes. The bare area of the sclerotic coat of the right eye was reconstructed with a latex biomembrane, and the corresponding site of the left eye was reconstructed with a conjunctival autograft. The animals were killed in groups of three at 7, 14, and 21 days after surgery. The tissues from the surgical site, including the cornea, were fixed in formaldehyde, and were then processed in paraffin and stained with hematoxylin and eosin. The nature and intensity of the inflammatory response and the epithelial pattern at the conjunctival surface were evaluated under optical microscopy with longitudinal histological sections through the center of the anatomical specimens. Results Until the 14th postoperative day, the inflammatory reaction was greater in the biomembrane group than in the conjunctival autograft group. In the latex biomembrane group, inflammation was less intense and the stroma was thicker on the 14th postoperative day than on the 7th postoperative day. After three weeks, conjunctival healing in both groups showed similar characteristics. Conclusion Although healing was slower with a latex biomembrane, tissue reconstitution was almost the same as that with a conjunctival autograft by three weeks. A latex biomembrane is as effective as a conjunctival autograft for the reconstruction of the ocular surface. Owing to the lack of toxicity and allergenicity, a latex biomembrane appears to be a promising therapeutic option for conjunctival reconstruction.

Arthropod venoms are sources of molecules that may be useful tools to investigate molecular mechanisms of putative new medicines and laboratory drugs. Here we show the effects of the compound agelaiatoxin-8 (AVTx8), isolated from Agelaia vicina venom, on γ-aminobutyric acid (GABA) neurotransmission in rat brain synaptosomes. Analysis reveals that AvTx8 is composed by 14 amino acid residues with a molecular weight (MW) of 1567 Da. AvTx8 increased GABA release and inhibited GABA uptake in synaptosomes from rat cerebral cortex. AvTx8 inhibited GABA uptake and increased GABA release in the presence of Ca+, Na+, and K+ channel blockers, suggesting that it acts directly on GABA transporters. In addition, AvTx8 significantly decreases GABA binding in synaptic membranes from rat brain cortex, suggesting that it also modulates the activity of GABA receptors. Moreover, AvTx8 decreased GAT-1– and GAT-3–mediated GABA uptake in transfected COS-7 cells. Accordingly, we suggest that AvTx8 modulates GABA neurotransmission and might provide a novel entry point for identifying a new class of GABA-modulating neuroprotective drugs.

The effects of low-level laser therapy (LLLT) and natural latex protein (F1, Hevea brasiliensis) were evaluated on crush-type injuries (15kg) to the sciatic nerve in the expressions of nerve growth factor (NGF) and vascular endothelium growth factor (VEGF) and ultrastructural morphology to associate with previous morphometric data using the same protocol of injury and treatment. Thirty-six male rats were allocated into six experimental groups (n = 6): 1-Control; 2-Exposed nerve; 3-Injured nerve; 4-LLLT (15J/cm2, 780nm, 30mW, Continuous Wave) treated injured nerve; 5-F1 (0,1mg) treated injured nerve; and 6-LLLT&F1 treated injured nerve. Four or eight weeks after, sciatic nerve samples were processed for analysis. NGF expression were higher (p

Caramboxin: Patients suffering from chronic kidney disease are frequently intoxicated after ingesting star fruit. The main symptoms of this intoxication are named in the picture. Bioguided chemical procedures resulted in the discovery of caramboxin, which is a phenylalanine-like molecule that is responsible for intoxication. Functional experiments in vivo and in vitro point towards the glutamatergic ionotropic molecular actions of caramboxin, which explains its convulsant and neurodegenerative properties.

The study of venom components is an important step toward understanding the mechanism of action of such venoms and is indispensable for the development of new therapies. This work aimed to investigate the venom of Lachesis muta rhombeata and evaluate enzymes related to its toxicity. Phospholipase A2 (PLA(2)), L-amino acid oxidase (LAAO), and proteinase activities were measured, and the molecular weights were estimated. We found the venom to contain one PLA(2) (17 kDa), one LAAO (132 kDa), and three serine proteinases (40, 31, and 20 kDa). Although only serine proteinases were observed in the zymogram, metalloproteinases were found to contribute more to the total proteolytic activity than did serine proteinases. The work confirmed the presence of highly active enzymes; and, moreover, we proposed a novel method for confirming the presence of LAAOs by zymography. We also suggested a simple step to increase the sensitivity of proteinase assays.

BACKGROUND: The vegetal biomembrane has been used to treat cutaneous ulcers. OBJECTIVES: To assess the role of the vegetal biomembrane on the chronic venous ulcers treatment compared to treatment with collagenase cream. METHODS: Fourteen patients were selected to be treated with vegetal biomembrane and 7 with Fibrase®(CONTROL), followed clinically and photographically by the Wound Healing Index by ImageJ during 120 days and biopsied on the 1st and 30th days for histological examination. RESULTS: The vegetal biomembrane was better in promoting healing of the ulcers, especially on the inflammatory phase, confirmed by abundant exudation and wound debridement than the CONTROL group, on the 30th day. There was a greater tendency to angiogenesis followed by re-epithelialization with highest wound healing index on the 90th and 120th days. CONCLUSION: A combined analysis of clinical and histopathological findings suggests that the vegetal biomembrane acted as a factor inducing wound healing, especially on the inflammatory phase, confirmed by abundant exudation of the lesions promoting the transformation of the microenvironment of the chronic venous ulcers, and also stimulating angiogenesis and subsequent re-epithelialization. FUNDAMENTOS: A biomembrana vegetal tem sido usada para tratamento de úlceras cutâneas. OBJETIVOS: Avaliar a ação da biomembrana vegetal no tratamento de úlceras venosas crônicas, comparando-a ao tratamento à base de colagenase. MÉTODOS: Foram selecionados 14 pacientes tratados com biomembrana vegetal e sete com Fibrase® (grupo controle), acompanhados clínico-fotograficamente pelo índice de cicatrização das úlceras (ICU) por 120 dias, por meio do software ImageJ, e biopsiados no primeiro e 30º dias para estudo histopatológico. RESULTADOS: A biomembrana vegetal foi superior em relação ao controle na cicatrização das úlceras no 30º dia, especialmente na fase inflamatória, confirmada pela exsudação abundante e pelo desbridamento. Houve tendência superior à angiogênese seguida de reepitelização com maiores ICUs no 90º e 120º dias. CONCLUSÃO: A análise conjunta dos achados clínicos e histopatológicos sugere que a biomembrana vegetal atuou como um fator indutor da cicatrização, especialmente na fase inflamatória, confirmada pela exsudação abundante das lesões, promovendo a transformação do microambiente das úlceras venosas crônicas e estimulando a angiogênese e a posterior reepitelização.

BACKGROUND: The vegetal biomembrane has been used to treat cutaneous ulcers. OBJECTIVES: To assess the role of the vegetal biomembrane on the chronic venous ulcers treatment compared to treatment with collagenase cream. METHODS: Fourteen patients were selected to be treated with vegetal biomembrane and 7 with Fibrase®(CONTROL), followed clinically and photographically by the Wound Healing Index by ImageJ during 120 days and biopsied on the 1st and 30th days for histological examination. RESULTS: The vegetal biomembrane was better in promoting healing of the ulcers, especially on the inflammatory phase, confirmed by abundant exudation and wound debridement than the CONTROL group, on the 30th day. There was a greater tendency to angiogenesis followed by re-epithelialization with highest wound healing index on the 90th and 120th days. CONCLUSION: A combined analysis of clinical and histopathological findings suggests that the vegetal biomembrane acted as a factor inducing wound healing, especially on the inflammatory phase, confirmed by abundant exudation of the lesions promoting the transformation of the microenvironment of the chronic venous ulcers, and also stimulating angiogenesis and subsequent re-epithelialization.FUNDAMENTOS: A biomembrana vegetal tem sido usada para tratamento de úlceras cutâneas. OBJETIVOS: Avaliar a ação da biomembrana vegetal no tratamento de úlceras venosas crônicas, comparando-a ao tratamento à base de colagenase. MÉTODOS: Foram selecionados 14 pacientes tratados com biomembrana vegetal e sete com Fibrase® (grupo controle), acompanhados clínico-fotograficamente pelo índice de cicatrização das úlceras (ICU) por 120 dias, por meio do software ImageJ, e biopsiados no primeiro e 30º dias para estudo histopatológico. RESULTADOS: A biomembrana vegetal foi superior em relação ao controle na cicatrização das úlceras no 30º dia, especialmente na fase inflamatória, confirmada pela exsudação abundante e pelo desbridamento. Houve tendência superior à angiogênese seguida de reepitelização com maiores ICUs no 90º e 120º dias. CONCLUSÃO: A análise conjunta dos achados clínicos e histopatológicos sugere que a biomembrana vegetal atuou como um fator indutor da cicatrização, especialmente na fase inflamatória, confirmada pela exsudação abundante das lesões, promovendo a transformação do microambiente das úlceras venosas crônicas e estimulando a angiogênese e a posterior reepitelização.

FUNDAMENTOS: A biomembrana vegetal do látex da seringueira Hevea brasiliensis tem sido usada como curativo para úlceras cutâneas. OBJETIVOS: Avaliar a segurança da biomembrana vegetal como curativo em relação à hipersensibilidade ao látex. MÉTODOS: Foram selecionados pacientes com úlceras cutâneas constituindo-se os grupos: controle - baixa exposição profissional ao látex (n=17); alta exposição profissional (n=14); ulcerados em uso da biomembrana vegetal (n=13); ulcerados-controle sem uso da biomembrana vegetal (n=14) e casos novos (n=9), submetidos à avaliação pré e após 3 meses de uso da biomembrana vegetal. Todos foram submetidos à avaliação clínico-epidemiológica quanto à hipersensibilidade ao látex e IgE específica (UniCap®), e os grupos controle e controle exposto ao látex ao "patch test". RESULTADOS: A história de hipersensibilidade foi positiva em 64,7% dos pacientes do grupo-controle, 71,4% do controle exposto ao látex, 61,5% dos ulcerados em uso da biomembrana vegetal, 35,7% dos ulcerados-controle, e apenas 22,2% no grupo casos novos. Ao teste de contato dos grupos controle e controle exposto ao látex, apenas um indivíduo do grupo C (baixo contato) apresentou eritema na primeira leitura, negativando-se na segunda. A média de contato com látex no grupo-controle exposto ao látex foi de 3,42 horas/dia. No ensaio fluoroimunoenzimático, a grande maioria dos soros foi classificada como zero (variação 0 a 6). Nenhum soro recebeu classificação acima de 2, não sendo considerada classificação significante para hipersensibilidade (classificação > 4). CONCLUSÃO: A biomembrana vegetal mostrou-se segura como curativo, pois não induziu reações de hipersensibilidade entre os voluntários submetidos ao "patch test", nem entre os usuários da biomembrana vegetal, como demonstrado clinica e imunologicamente pela dosagem de IgE.BACKGROUND: The natural biomembrane of latex extracted from Hevea brasiliensis has been used as a dressing for skin ulcers. OBJECTIVES: To evaluate how safe the natural biomembrane is in relation to hypersensitivity to latex when used as a dressing. METHODS: We selected patients with skin ulcers, forming the following groups: control - low occupational exposure to latex (n = 17); latex-exposed control - high occupational exposure (n = 14); ulcerated, using the natural biomembrane (n = 13); ulcerated control, not using the natural biomembrane (n = 14); and new cases (n = 9), assessed before and after 3 months of using the natural biomembrane. All patients underwent clinical and epidemiological evaluation for latex hypersensitivity and specific IgE (UniCap®), and the control and latex-exposed control groups underwent the patch test. RESULTS: Hypersensitivity was positive in 64.7% of the patients in the control group, 71.4% of the patients in the latex-exposed control group, 61.5% of the ulcerated using the natural biomembrane, 35.7% of the ulcerated control, and only 22 , 2% of the new cases. In the patch test of the control and latex-exposed control groups, only one individual in the control group (low contact) showed erythema in the first reading, which became negative in the second. The mean contact with latex in the latex-exposed control group was 3.42 hours / day. In the fluoroimmunoenzymatic assay, most of the sera was classified as zero (range 0-6). No serum was rated above 2, which is not considered significant for hypersensitivity (classification > 4). CONCLUSION: The natural biomembrane proved to be safe as a dressing, for it did not induce hypersensitivity reactions among the volunteers who underwent the patch test or among users of the natural biomembrane, as it was clinically and immunologically demonstrated by IgE levels.

Background: The aim of this work was to study the new bone tissue formation af- ter bone morphogenetic protein type 2 (rhBMP-2) and P-1 application, using 5 and 10 lg of each, combined to a material carrier, in critical bone defects. Methods: It was used 70 Wistar rats (male, 250 g) that were divided in 10 groups with seven animals on each. Groups are the following: criti- cal bone defect only, pure monoolein gel, 5 lg of pure P-1, 5 lg of pure rhBMP-2, 5 lg of P-1/monoo- lein gel, 5 lg of rhBMP-2/monoolein gel, 10 lg of pure P-1, 10 lg of pure rhBMP-2, 10 lg of P-1/ monoolein gel, 10 lg of rhBMP-2/monoolein gel. Animals were sacrificed after 4 weeks of the surgi- cal procedure and the bone samples were submitted to histological, histomorphometrical, and im- munohistochemical evaluations. Results: Animals treated with pure P-1 protein, in both situations with 5 lg and 10 lg, had no significant difference (P > 0.05) for new bone formation; other groups treated with 10 lg were statistically significant (P < 0.05) among themselves and when compared with groups in which it was inserted the monoolein gel or critical bone defect only (P < 0.05). In the group involving the 10 lg rhBMP-2/monoolein gel association, it was observed an extensive bone formation, even when compared with the same treatment without the gel carrier. Conclusion: Using this experimental animal model, more new bone tissue was found when it was inserted the rhBMP-2, especially when this protein was combined to the vehicle, and this process seems to be dose dependent. Microsc. Res. Tech. 00:000-000, 2011. V C 2011 Wiley-Liss, Inc.

PURPOSE: To evaluate the morphological aspects of the behavior of 4 types of latex biomembranes implanted in preperitoneal videolaparoscopic inguinoplasty. METHODS: Sixteen inguinoplasties were performed in 12 dogs: group 1 received an impermeable latex biomembrane in the right inguinal region and a prolene prosthesis, as control, in the contralateral inguinal region; groups 2, 3 and 4 received latex biomembranes respectively containing impermeable polyamide, 1-mm thick porous polyamide and 0.5-mm thick porous polyamide. Macro- and microscopic evaluations of the inguinal region and of the removed implants were made on the 7th, 14th, 21st and 28th days in group 1 and on the 28th postoperative day in the other groups. RESULTS: We observed absence of hematoma, seroma and infection; presence of tortuosities; induction of vascular neoformation, inflammatory reaction and collagen deposition, and full encystment of the latex biomembranes, except that with fine porous polyamide, which was partially incorporated, with the formation of microcysts. No latex biomembrane induced fibrosis as observed in the prolene control group. CONCLUSIONS: The biomembranes maintain induction of the healing process without fibrosis, are fully encysted and, except for the one with fine porous polyamide, are not incorporated into adjacent tissues. The latex biomembrane, with or without polyamide, is not recommended as a separate material for preperitoneal inguinoplasty.OBJETIVO: Avaliar aspectos morfológicos do comportamento de 4 tipos de biomembranas de latex, colocadas pré-peritonealmente em cães, por inguinoplastia videolaparoscópica. MÉTODOS: Dezesseis inguinoplastias em 12 cães: grupo 1, com biomembrana de latex impermeável inguinal direita em quatro cães e prótese de prolene, como controle, contra-lateral; grupos 2, 3 e 4, com biomembrana de latex respectivamente de poliamida impermeável, poliamida porosa com 1mm de espessura e poliamida porosa com 0,5mm de espessura. Avaliou-se a região inguinal e as peças retiradas macro e microscopicamente, no grupo 1 no 7º, 14º, 21º e 28º dias e nos demais grupos no 28º dia pós-operatório. RESULTADOS: ausência de hematoma, seroma e infecção; presença de tortuosidade; indução de neoformação vascular, reação inflamatória, deposição de colágeno e encistamento total das biomembranas de latex, exceto com poliamida porosa fina que se incorporou apenas parcialmente, com formação de microcistos. Nenhuma biomembrana de latex induziu fibrose como no grupo controle prolene. CONCLUSÕES: As biomembranas mantêm indução do processo de cicatrização sem fibrose, sofrem encistamento e, exceto com poliamida porosa fina, não se incorporam aos tecidos vizinhos. A biomembrana de latex, com e sem poliamida, isoladamente não é recomendada para inguinoplastia pré-peritoneal.

OBJETIVO: Avaliar os aspectos morfológicos do comportamento de prótese de dupla face aplicada em inguinoplastia laparotômica em cães, com fixação intraperitoneal com a face de látex voltada às vísceras. MÉTODOS: Vinte cães distribuídos em dois grupos (n=10) foram submetidos à laparotomia infraumbilical com fixação da prótese de dupla face em uma região inguinal e de uma prótese controle de polipropileno contralateral. Foram pesquisados no 14° e 28° dia de pós-operatório achados macroscópicos referentes à obstrução e fístula intestinais, encistamento, incorporação e aderências. A análise microscópica envolveu o processo inflamatório e reparador. RESULTADOS: Não ocorreram processos infecciosos, obstrução ou fístula intestinal. As próteses apresentaram boa acomodação e incorporação. As aderências ocorreram em maior prevalência e intensidade com a prótese de polipropileno (p0,05). CONCLUSÃO: A prótese de dupla face na sua face parietal soma as vantagens do potencial de incorporação aos tecidos observados com o polipropileno às de biocompatibilidade do látex na sua face visceral. A pequena distância entre o disco de polipropileno e a borda da prótese de dupla face (2 cm) aliada à sua fixação com apenas cinco grampos é insuficiente para evitar que o epíploon migre em direção ao processo inflamatório desencadeado pelo polipropileno na face parietal.

Department of Oral and Maxillofacial Surgery and Periodontics Faculty of Dentistry of Ribeirao Preto University of Sao Paulo, Ribeirao Preto, Sao Paulo

Objectives The purpose of the present work was to characterize the pharmacological profile of different L. alba chemotypes and to correlate the obtained data to the presence of chemical constituents detected by phytochemical analysis. Methods Essential oils from each L. alba chemotype (LP1—LP7) were characterized by gas chromatography–mass spectrometry (GC-MS) and extracted non-volatile compounds were analysed by HPLC and GC-MS. The anticonvulsant actions of the extracted compounds were studied in pentylenetetrazole-induced clonic seizures in mice and their effect on motor coordination was studied using the rota-rod test in rats. The synaptosomes and synaptic membranes of the rats were examined for the influence of LP3 chemotype extract on GABA uptake and binding experiments. Key findings Behavioural parameters encompassed by the pentylenetetrazole test indicated that 80% ethanolic extracts of LP1, LP3 and LP6 L. alba chemotypes were more effective as anticonvulsant agents. Neurochemical assays using synaptosomes and synaptic membranes showed that L. alba LP3 chemotype 80% ethanolic extract inhibited GABA uptake and GABA binding in a dose-dependent manner. HPLC analysis showed that LP1, LP3 and LP6 80% ethanolic extracts presented a similar profile of constituents, differing from those seen in LP2, LP4, LP5 and LP7 80% ethanolic extracts, which exhibited no anticonvulsant effect. GC-MS analysis indicated the occurrence of phenylpropanoids in methanolic fractions obtained from LP1, LP3 and LP6 80% ethanolic extracts and also the accumulation of inositol and flavonoids in hydroalcoholic fractions. Conclusions Our results suggest that the anticonvulsant properties shown by L. alba might be correlated to the presence of a complex of non-volatile substances (phenylpropanoids, flavonoids and/or inositols), and also to the volatile terpenoids (β-myrcene, citral, limonene and carvone), which have been previously validated as anticonvulsants.

OBJETIVO: avaliar a utilização do molde de látex natural (Hevea brasiliensis) como modificação à neovaginoplastia de McIndoe e Bannister em pacientes portadoras da síndrome de Mayer-Rokitansky-Küster-Hauser (MRKH). MÉTODOS: análise retrospectiva de nove pacientes com o diagnóstico de síndrome de MRKH, submetidas à neovaginoplastia pela técnica de McIndoe e Bannister com molde de látex natural. Foram avaliadas epitelização, amplitude e profundidade das neovaginas, ocorrência de coitos bem como satisfação e complicações cirúrgicas. RESULTADOS: após cinco semanas do procedimento, oito pacientes apresentavam neovaginas epitelizadas, com profundidade de 7 a 12 cm. Houve um caso de estenose neovaginal completa em decorrência do uso incorreto do molde pela paciente no pós-operatório. Após seguimento mínimo de um ano, todas as pacientes mantinham neovaginas com profundidade de 4 a 8 cm e capacidade para o coito, com 66,7% de satisfação. Uma paciente apresentou fístula retovaginal precoce e episódios tardios de fistulização uretrovaginal. Duas pacientes apresentaram estenose distal das neovaginas a longo prazo. Uma delas e a paciente com fístulas foram submetidas a novo procedimento. CONCLUSÕES: o uso do molde de látex natural como modificação à técnica clássica de neovaginoplastia permitiu a criação de neovaginas morfológica e funcionalmente similares ao órgão normal em pacientes com estenose vaginal.PURPOSE: to evaluate the use of natural latex mold (Hevea brasiliensis) as a modification of McIndoe and Bannister neovaginoplasty in patients presenting Mayer-Rokitansky-Küster-Hauser (MKRH) syndrome. METHODS: we retrospectively included nine patients presenting MKRH syndrome, who had been submitted to McIndoe and Bannister neovaginoplasty modified by the use of natural latex mold. Neovaginal epithelization and depth, coitus occurrence and satisfaction, and surgical complications were evaluated. RESULTS: five weeks after the procedure, eight patients presented an epithelized 7 to 12 cm deep neovagina. There was one case of complete neovaginal stenosis, because of incorrect use of the mold. After at least one year, the others maintained 4 to 8 cm deep neovaginas and capacity for intercourse, with 66.7% satisfaction. One woman presented precocious rectovaginal fistula and late episodes of uretrovaginal fistulae. Two patients presented distal neovaginal stenosis in long-term follow-up. One of these and the patient with fistulae were submitted to a new procedure. CONCLUSIONS: the use of natural latex mold as a modification of classic neovaginoplasty technique allows the creation of neovaginas morphologically and functionally similar to the normal vagina in patients with vaginal agenesis.

OBJECTIVE: To study the neurobehavioral, biochemical and histopathological consequences of permanent focal brain ischemia, and the putative neuroprotective action of ketoprofen. METHOD: One-hundred-and-three Wistar rats divided into groups A and B were respectively submitted to 48 hours and 15 days of ischemia. Each group was divided into 4 subgroups: ischemic not treated, ischemic treated, sham not treated, and sham treated. Ischemic animals had the left middle cerebral artery coagulated. Ketoprofen was administered to treated subgroups 15 minutes before arterial coagulation (manipulation in the sham group). RESULTS: Exploratory activity and defecation were reduced in all ischemic animals in the first postoperative days and constant histopathological changes were observed in each group. The total brain glutamate levels were higher in treated animals 48 hours after surgery. CONCLUSION: No clear parallelism among behavioral, biochemical and histopathological findings was observed. Ketoprofen demonstrated no neuroprotective effect on the behavioral or histopathological aspects of focal permanent brain ischemia.OBJETIVO: Estudar as conseqüências comportamentais, bioquímicas e histopatológicas da isquemia cerebral focal permanente e o possível efeito neuroprotetor do cetoprofeno. MÉTODO: Foram utilizados 103 ratos Wistar, divididos em grupos A e B, submetidos, respectivamente, a 48 horas e a 15 dias de isquemia. Cada grupo foi dividido em 4 subgrupos: isquêmico não tratado; isquêmico tratado; sham não tratado; sham tratado. Nos animais isquêmicos foi coagulada a artéria cerebral média esquerda. Os subgrupos tratados receberam cetoprofeno 15 minutos antes da oclusão ou manipulação arterial. RESULTADOS: Os animais isquêmicos reduziram a atividade exploratória e as evacuações nos primeiros dias pós-operatórios e mostraram alterações histopatológicas constantes em cada grupo. As concentrações do glutamato total 48 horas após a cirurgia foram maiores nos animais tratados. CONCLUSÃO: Não houve um paralelismo entre os achados comportamentais, bioquímicos e histopatológicos. O cetoprofeno não apresentou efeito protetor contra isquemia cerebral focal permanente, nos aspectos comportamentais e histopatológicos.

Previous studies have shown that a compound purified from the spider Parawixia bistriata venom stimulates the activity of glial glutamate transporters and can protect retinal tissue from ischemic damage. To understand the mechanism by which this compound enhances transport, we examined its effects on the functional properties of glutamate transporters after solubilization and reconstitution in liposomes and in transfected COS-7 cells. Here, we demonstrate in both systems that Parawixin1 promotes a direct and selective enhancement of glutamate influx by the EAAT2 transporter subtype through a mechanism that does not alter the apparent affinities for the cosubstrates glutamate or sodium. In liposomes, we observed maximal enhancement by Parawixin1 when extracellular sodium and intracellular potassium concentrations are within physiological ranges. Moreover, the compound does not enhance the reverse transport of glutamate under ionic conditions that favor efflux, when extracellular potassium is elevated and the sodium gradient is reduced, nor does it alter the exchange of glutamate in the absence of internal potassium. These observations suggest that Parawixin1 facilitates the reorientation of the potassium-bound transporter, the rate-limiting step in the transport cycle, a conclusion further supported by experiments showing that Parawixin1 does not stimulate uptake by an EAAT2 transport mutant (E405D) defective in the potassium-dependent reorientation step. Thus, Parawixin1 enhances transport through a novel mechanism targeting a step in the transport cycle distinct from substrate influx or efflux and provides a basis for the design of new drugs that act allosterically on transporters to increase glutamate clearance.

Background and purpose: In this work, a neuroactive peptide from the venom of the neotropical wasp Polybia occidentalis was isolated and its anti-nociceptive effects were characterized in well-established pain induction models. Experimental approach: Wasp venom was analysed by reverse-phase HPLC and fractions screened for anti-nociceptive activity. The structure of the most active fraction was identified by electron-spray mass spectrometry (ESI-MS/MS) and it was further assessed in two tests of anti-nociceptive activity in rats: the hot plate and tail flick tests. Key Results: The most active fraction contained a peptide whose structure was Arg-Pro-Pro-Gly-Phe-Thr-Pro-Phe-Arg-OH, which corresponds to that of Thr6-BK, a bradykinin analogue. This peptide was given by i.c.v. injection to rats. In the tail flick test, Thr6-BK induced anti-nociceptive effects, approximately twice as potent as either morphine or bradykinin also given i.c.v. The anti-nociceptive activity of Thr6-BK peaked at 30 min after injection and persisted for 2 h, longer than bradykinin. The primary mode of action of Thr6-BK involved the activation of B2 bradykinin receptors, as anti-nociceptive effects of Thr6-BK were antagonized by a selective B2 receptor antagonist. Conclusions and implications: Our data indicate that Thr6-BK acts through B2 bradykinin receptors in the mammalian CNS, evoking antinociceptive behaviour. This activity is remarkably different from that of bradykinin, despite the structural similarities between both peptides. In addition, due to the increased metabolic stability of Thr6-BK, relative to that of bradykinin, this peptide could provide a novel tool in the investigation of kinin pathways involved with pain. British Journal of Pharmacology (2007) 151, 860–869; doi:10.1038/sj.bjp.0707275

CONTEXTO: O desenvolvimento de prótese vascular tem sido vital para os avanços e realizações da cirurgia vascular reconstrutora durante as últimas 5 décadas. OBJETIVOS: Desenvolver um novo modelo de prótese vascular microperfurada, confeccionada em tecido recoberto com um composto derivado do látex natural da seringueira (Hevea brasiliensis) e avaliar sua perviedade, trombogenicidade, biocompatibilidade e o processo de cicatrização, além de algumas propriedades mecânicas (adaptabilidade, elasticidade, impermeabilidade e possibilidade de sutura), utilizando como controle a prótese de politetrafluoretileno expandido no mesmo animal. MÉTODOS: Quinze cães foram separados em três grupos de cinco animais. Implantou-se a prótese de tecido e látex microperfurada e, no membro pélvico contralateral, a prótese de politetrafluoretileno expandido em todos os cães. O seguimento pós-operatório foi de 4, 8 e 12 semanas. A apreciação dos resultados foi feita segundo as avaliações clínicas dos pulsos, complicações (coleção líquida, deiscência, granuloma e infecção), arteriografias, análise macroscópica e elétron-micrografias de varredura. RESULTADOS: Os testes estatísticos aplicados não evidenciaram diferenças significativas (p > 0,05) em relação às complicações pós-operatórias e perviedade dos enxertos. Ambas as próteses integraram-se adequadamente aos tecidos circunvizinhos, com um tecido de incorporação formado por fibras colágenas. Constatou-se a presença de neoíntima recoberta por endotélio em toda a extensão da superfície luminal da prótese de tecido e látex microperfurada. Ao contrário, na prótese de politetrafluoretileno expandido, o desenvolvimento endotelial sobre a superfície neointimal limitou-se às regiões próximas às anastomoses. CONCLUSÕES: A prótese de tecido e látex microperfurada demonstrou qualidades estruturais (adaptabilidade, elasticidade, impermeabilidade e possibilidade de sutura) satisfatórias como substituto vascular. Estimulou o crescimento endotelial além das regiões de contato com a artéria nas anastomoses e foi biocompatível no sistema arterial do cão, apresentando adequada integração tecidual.BACKGROUND: The development of vascular grafts has been crucial for advances and achievements in reconstructive vascular surgery over the past 5 decades. OBJECTIVES: To develop a new model of microperforated vascular graft using fabric covered with a natural latex-derived polymer taken from Hevea brasiliensis and assess its patency, thrombogenicity, biocompatibility and healing process, besides some mechanical properties (adaptability, elasticity, impermeability and possibility of suture), using expanded polytetrafluoroethylene graft as control. METHODS: Fifteen dogs were divided into three groups of five animals. The microperforated latex graft was implanted in all dogs and the expanded polytetrafluoroethylene graft was implanted in the contralateral pelvic limb. Postoperative follow-up was 4, 8 and 12 weeks. Analysis of results was performed according to clinical evaluation of pulses, complications (fluid collection, dehiscence, granuloma and infection), arteriography, macroscopic analysis and scanning electron micrography. RESULTS: Statistical tests revealed no significant differences (p > 0.05) concerning post-operative complications and graft patency. Both grafts were properly integrated to surrounding tissues, with connective tissue formed by collagen fibers. A neointimal layer covering all extension of the luminal surface was observed in the microperforated latex graft. Conversely, the endothelial development over the neointimal surface was limited to regions adjacent to the anastomoses in the expanded polytetrafluoroethylene graft. CONCLUSIONS: The microperforated latex graft showed satisfactory structural qualities (adaptability, elasticity, impermeability and possibility of suture) as a vascular substitute. It stimulated endothelial growth beyond contact regions with the artery in anastomoses and was biocompatible in the dog's arterial system, presenting adequate tissue integration.

As neocavidades pós-timpanomastoidectomias de cavidade aberta (TMCA) são normalmente preenchidas por cadarço ou fita cardíaca untados com pomada antibiótica. A remoção deste tampão usualmente causa sangramento e desconforto para o paciente. Propomos para tanto a utilização de uma biomembrana de látex natural para forrar a neocavidade, servindo como uma interface entre o osso cruento e o material de tamponamento. FORMA DE ESTUDO: Clínico prospectivo. OBJETIVOS: Estudar o desempenho da biomembrana como uma interface entre o osso cruento e o material de tamponamento e analisar seu papel na epitelização da neocavidade. MATERIAL E MÉTODO: Foram analisadas 64 orelhas de pacientes submetidos à TMCA e meatoplastia. A biomembrana foi utilizada em 54 das orelhas operadas, sendo que nas outras 10 orelhas o tamponamento da cavidade foi realizado somente com fita cardíaca. RESULTADOS: Na maioria das 54 orelhas onde a biomembrana foi utilizada observou-se maior facilidade na remoção do curativo tampão (sem sangramento ou desconforto para o paciente), menor demanda de tempo para sua retirada, além da epitelização mais precoce da neocavidade. CONCLUSÃO: A utilização da biomembrana de látex revelou-se método eficaz no revestimento da neocavidade, facilitando a remoção do tampão e a epitelização da neocavidade.

As neocavidades pós-timpanomastoidectomias de cavidade aberta (TMCA) são normalmente preenchidas por cadarço ou fita cardíaca untados com pomada antibiótica. A remoção deste tampão usualmente causa sangramento e desconforto para o paciente. Propomos para tanto a utilização de uma biomembrana de látex natural para forrar a neocavidade, servindo como uma interface entre o osso cruento e o material de tamponamento. FORMA DE ESTUDO: Clínico prospectivo. OBJETIVOS: Estudar o desempenho da biomembrana como uma interface entre o osso cruento e o material de tamponamento e analisar seu papel na epitelização da neocavidade. MATERIAL E MÉTODO: Foram analisadas 64 orelhas de pacientes submetidos à TMCA e meatoplastia. A biomembrana foi utilizada em 54 das orelhas operadas, sendo que nas outras 10 orelhas o tamponamento da cavidade foi realizado somente com fita cardíaca. RESULTADOS: Na maioria das 54 orelhas onde a biomembrana foi utilizada observou-se maior facilidade na remoção do curativo tampão (sem sangramento ou desconforto para o paciente), menor demanda de tempo para sua retirada, além da epitelização mais precoce da neocavidade. CONCLUSÃO: A utilização da biomembrana de látex revelou-se método eficaz no revestimento da neocavidade, facilitando a remoção do tampão e a epitelização da neocavidade.The new cavity created after an open cavity tympanomastoidectomy (OCTM) is filled with an antibiotic impregnated cotton pack (cotton tape, umbilical tape, gauze). The removal of this pack usually causes some bleeding and discomfort for the patient. We propose the use of a latex biomembrane to cover the cavity, which will act as an interface between the raw bone surface and the packing. STUDY DESIGN: clinical prospective. AIM: To study the performance of the latex biomembrane as an interface between the raw bone surface and the pack, and to analyze its role in cavity epithelization. MATERIAL AND METHODS: 64 ears of patients submitted to OCTM were studied. The biomembrane was used in the packing of 54 ears and in the 10 remaining ears the regular cotton tape packing was used. RESULTS: In the majority of the cases where the biomembrane was used the packing was removed much easier with no bleeding or pain for the patient and also showed an earlier cavity epithelization. CONCLUSION: The use of the latex biomembrane has proven to be an effective method to cover the mastoid cavity facilitating epithelization and removal of mastoid cavity packing.

Summary The new cavity created after an open cavity tympanomastoidectomy (OCTM) is filled with an antibiotic impregnated cotton pack (cotton tape, umbilical tape, gauze). The removal of this pack usually causes some bleeding and discomfort for the patient. We propose the use of a latex biomembrane to cover the cavity, which will act as an interface between the raw bone surface and the packing. Study design: clinical prospective. Aim: To study the performance of the latex biomembrane as an interface between the raw bone surface and the pack, and to analyze its role in cavity epithelization. Material and Methods: 64 ears of patients submitted to OCTM were studied. The biomembrane was used in the packing of 54 ears and in the 10 remaining ears the regular cotton tape packing was used. Results: In the majority of the cases where the biomembrane was used the packing was removed much easier with no bleeding or pain for the patient and also showed an earlier cavity epithelization. Conclusion: The use of the latex biomembrane has proven to be an effective method to cover the mastoid cavity facilitating epithelization and removal of mastoid cavity packing.

This study was aimed at determining the effects of FrPbAII (174 Da), a novel isolated component from Parawixia bistriata spider venom, in the CNS of Wistar rats. Considering that FrPbAII inhibits the high affinity GABAergic uptake in a dose-dependent manner, its anxiolytic and anticonvulsant effects were analyzed in well-established animal models. Injection of FrPbAII in the rat hippocampus induced a marked anxiolytic effect, increasing the occupancy in the open arms of the elevated plus maze (EC(50)=0.09 microg/microl) and increasing the time spent in the lit area of the light-dark apparatus (EC(50)=0.03 microg/microl). Anxiolytic effects were also observed considering the number of entries in the open arms of the EPM and in the lit compartment of the light-dark box. Interestingly, when microinjected bilaterally in the SNPr of freely moving rats, FrPbAII (0.6 microg/microl) effectively prevented seizures induced by the unilateral GABAergic blockade of Area tempestas (bicuculline, 0.75 microg/microl). This anticonvulsant effect was similar to that evoked by muscimol (0.1 microg/microl) and baclofen (0.6 microg/microl), but differed from that of the specific GAT1 inhibitor, nipecotic acid (0.7 microg/microl). This difference could be accounted either for the parallel action of FrPbAII over glycinergic transporters or to an inspecific activity on GABAergic transporters. Data from the present investigation might be pointing to a novel compound with interesting and yet unexplored pharmacological potential.

The major contribution of this work is the isolation of a neuroprotective compound referred to as 2-amino-5-ureidopentanamide (FrPbAII) (M(r) = 174) from Parawixia bistriata spider venom and an investigation of its mode of action. FrPbAII inhibits synaptosomal GABA uptake in a dose-dependent manner and probably does not act on Na(+), K(+), and Ca(2+) channels, GABA(B) receptors, or gamma-aminobutyrate:alpha-ketoglutarate aminotransferase enzyme; therefore, it is not directly dependent on these structures for its action. Direct increase of GABA release and reverse transport are also ruled out as mechanisms of FrPbAII activities as well as unspecific actions on pore membrane formation. Moreover, FrPbAII is selective for GABA and glycine transporters, having slight or no effect on monoamines or glutamate transporters. According to our experimental glaucoma data in rat retina, FrPbAII is able to cross the blood-retina barrier and promote effective protection of retinal layers submitted to ischemic conditions. These studies are of relevance by providing a better understanding of neurochemical mechanisms involved in brain function and for possible development of new neuropharmacological and therapeutic tools.

The mechanical properties of rubber tubes produced by dip-coating technique and the influence of the fabrication process on their structural properties were evaluated. Cyclic tension versus deformation tests were performed to investigate the elastic properties of the samples and to understand the changes in rubber tubes behavior under repetitive stress. The effect of the wall thickness on the elastic response of the tubes was also studied. The mechanical properties of opened tubes were also investigated for transversal and longitudinal directions to evaluate the influence of the fabrication process on the alignment of the polymer chains. This investigation indicated that the fabricated tubes are resistant and extremely elastic. They can be elongated up to 800% of its initial length before rupture, and thicker tubes are a bit more resistant to elongation than the thinner ones. In addition, the fabricated tubes have an anisotropic structure due to the fabrication process. Finally, natural rubber tubes may have a great potential to be used as vascular prosthesis, or in other applications that require a large range of resistance and elasticity. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 702–707, 2006

We obtained a neurotoxic fraction (AcTx) from star fruit (Averrhoa carambola) and studied its effects on GABAergic and glutamatergic transmission systems. AcTx had no effect on GABA/glutamate uptake or release, or on glutamate binding. However, it specifically inhibited GABA binding in a concentration-dependent manner (IC50 = 0.89 mM). Video-electroencephalogram recordings demonstrated that following cortical administration of AcTx, animals showed behavioral changes, including tonic-clonic seizures, evolving into status epilepticus, accompanied by cortical epileptiform activity. Chemical characterization of AcTx showed that this compound is a nonproteic molecule with a molecular weight less than 500, differing from oxalic acid. This neurotoxic fraction of star fruit may be considered a new tool for neurochemical and neuroethological research. # 2005 Elsevier Ltd. All rights reserved.

Venoms of spiders and wasps are well recognized to present high affinity to the central nervous tissue of many mammalian species. Here we describe the effects of direct exposure of rat (Rattus norvegicus) brains to the crude and denatured venom of the Brazilian social wasp Polybia ignobilis. Lower doses of crude venom injected via intracerebroventricular (i.c.v.) inhibited the exploratory activity of animals, while higher doses provoked severe generalized tonic-clonic seizures, with hind limb extension. The status epilepticus lasted for few minutes leading the animals to respiratory depression and death. In contrast, the denatured venom was anticonvulsant against acute seizures induced by the i.c.v. injection of bicuculline, picrotoxin and kainic acid, but it was ineffective against seizures caused by systemic pentylenetetrazole. Moreover, the [3H]-glutamate binding in membranes from rat brain cortex was inhibited by the denatured venom in lower concentrations than the [3H]-GABA binding. The denatured venom contains free GABA and glutamate (34 and 802 pg/microg of venom, respectively), but they are not the major binding inhibitors. These interactions of venom components with GABA and glutamate receptors could be responsible for the anticonvulsant effects introducing the venom from P. ignobilis as a potential pharmacological source of anticonvulsant drugs.

INTRODUÇÃO: A biomembrana de látex natural foi utilizada com sucesso nas reconstruções do esôfago, da parede abdominal e do pericárdio de animais, em que puderam ser comprovadas a biocompatibilidade e a capacidade de favorecimento do reparo tecidual desse material. No homem, ela já está sendo testada como material indutor de neoformação tecidual, tendo sido aplicada em pacientes com úlceras crônicas de membros inferiores e meringoplastias. OBJETIVO: Verificar o efeito da biomembrana de látex no processo de reparo da conjuntiva ocular. MÉTODOS: Promoveu-se a retirada de retângulos da conjuntiva nasal superior, de ambos os olhos, de 15 coelhos neo-zelandeses adultos. Nos olhos direitos, foram implantadas biomembranas de látex com suturas contínuas presas às bordas das lesões cirúrgicas. Nos esquerdos, foram deixadas as escleras nuas. Para as análises histológicas, sacrificaram-se os animais com cinco, sete, catorze, vinte e um e vinte e oito dias. Os olhos de um coelho, não submetido a qualquer procedimento, foram usados como controle histológico. RESULTADOS: Considerando o período total de estudo, o grupo com biomembrana de látex apresentou cicatrização satisfatória em maior número de olhos do que o grupo com esclera nua (p=0,06). O número de vasos perilímbicos também foi significativamente maior nos casos com implante de biomembrana do que nos olhos sem biomembrana (p=0,0284). A freqüência de infecções foi idêntica nos dois grupos. CONCLUSÃO: Tal como o descrito na literatura para outros tecidos, a biomembrana de látex natural também parece favorecer a cicatrização conjuntival e a neoangiogênese. Se esses resultados se repetirem nos humanos, a biomembrana poderá se converter num promissor recurso terapêutico de reconstrução da conjuntiva ocular, particularmente nos casos em que a revascularização tecidual seja importante.INTRODUCTION: The biomembrane of natural latex is believed to promote neoformation of biological tissues and to be biocompatible. In animal studies, enhancement of wound healing in the esophagus, peritoniostomy, and pericardiostomy has been observed. It was used in chronic phlebopatic cutaneous ulcer and otologic surgeries in human subjects. PURPOSE: To study the effect of biomembrane of natural latex in ocular surface. METHODS: In fifteen New Zealand white rabbits, the superior nasal conjunctiva was removed from both eyes. In the right eyes, the biomembrane of natural latex was sutured to the conjunctiva with continuous 10.0 monofilament thread. In the left eye, the sclera was left bare. The rabbits were sacrified and the eyes were submitted to histological analysis on days five, seven, fourteen, twenty-one and twenty-eight. The eyes of a rabbit with no surgery were enucleated and sent for histological analysis of the normal conjunctiva. RESULTS: Considering the whole period of the experiment, the number of eyes with satisfactory healing was greater in the biomembrane group than in the bare sclera group (p=0.06). The number of perilimbic vessels was significantly higher in the biomembrane group than in the bare sclera group (p=0.0284). The infection rate was similar in both groups. CONCLUSION: Similar to prior studies, the biomembrane of natural latex seemed to benefit conjunctival reconstruction and neoangiogenesis. Depending on the results using human eyes, this material might be a new type of therapeutic resource for conjunctival replacement, especially in cases of desirable tissue revascularization.

It has previously been shown that the denatured crude extract of Agelaia vicina wasp venom inhibits glutamate and GABA uptake in rat cerebral cortex synaptosomes. To identify the components responsible for these effects, the neurotoxin AvTx 7 (molecular weight of 1210 Da) was isolated from A. vicina venom and its effects on glutamate neurotransmission investigated. AvTx 7 inhibits glutamate uptake in a dose-dependent and uncompetitive manner. AvTx 7 was found to stimulate the glutamate release in the presence of calcium and sodium channel blockers, suggesting that its action is not mediated through these channels. AvTx 7 potentiates glutamate release in the presence of K+ channel blockers tetraethylammonium and 4-aminopyridine, indicating that the toxin may act through these drugs-sensible K+ channels. We suggest that AvTx 7 can be a valuable tool to enhance our understanding of K+ channels' involvement in the release of glutamate. © 2004 Wiley Periodicals, Inc. J Biochem Mol Toxicol 18:61–68, 2004; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20014

INTRODUÇÃO: A biomembrana de látex natural foi utilizada com sucesso nas reconstruções do esôfago, da parede abdominal e do pericárdio de animais, em que puderam ser comprovadas a biocompatibilidade e a capacidade de favorecimento do reparo tecidual desse material. No homem, ela já está sendo testada como material indutor de neoformação tecidual, tendo sido aplicada em pacientes com úlceras crônicas de membros inferiores e meringoplastias. OBJETIVO: Verificar o efeito da biomembrana de látex no processo de reparo da conjuntiva ocular. MÉTODOS: Promoveu-se a retirada de retângulos da conjuntiva nasal superior, de ambos os olhos, de 15 coelhos neo-zelandeses adultos. Nos olhos direitos, foram implantadas biomembranas de látex com suturas contínuas presas às bordas das lesões cirúrgicas. Nos esquerdos, foram deixadas as escleras nuas. Para as análises histológicas, sacrificaram-se os animais com cinco, sete, catorze, vinte e um e vinte e oito dias. Os olhos de um coelho, não submetido a qualquer procedimento, foram usados como controle histológico. RESULTADOS: Considerando o período total de estudo, o grupo com biomembrana de látex apresentou cicatrização satisfatória em maior número de olhos do que o grupo com esclera nua (p=0,06). O número de vasos perilímbicos também foi significativamente maior nos casos com implante de biomembrana do que nos olhos sem biomembrana (p=0,0284). A freqüência de infecções foi idêntica nos dois grupos. CONCLUSÃO: Tal como o descrito na literatura para outros tecidos, a biomembrana de látex natural também parece favorecer a cicatrização conjuntival e a neoangiogênese. Se esses resultados se repetirem nos humanos, a biomembrana poderá se converter num promissor recurso terapêutico de reconstrução da conjuntiva ocular, particularmente nos casos em que a revascularização tecidual seja importante. INTRODUCTION: The biomembrane of natural latex is believed to promote neoformation of biological tissues and to be biocompatible. In animal studies, enhancement of wound healing in the esophagus, peritoniostomy, and pericardiostomy has been observed. It was used in chronic phlebopatic cutaneous ulcer and otologic surgeries in human subjects. PURPOSE: To study the effect of biomembrane of natural latex in ocular surface. METHODS: In fifteen New Zealand white rabbits, the superior nasal conjunctiva was removed from both eyes. In the right eyes, the biomembrane of natural latex was sutured to the conjunctiva with continuous 10.0 monofilament thread. In the left eye, the sclera was left bare. The rabbits were sacrified and the eyes were submitted to histological analysis on days five, seven, fourteen, twenty-one and twenty-eight. The eyes of a rabbit with no surgery were enucleated and sent for histological analysis of the normal conjunctiva. RESULTS: Considering the whole period of the experiment, the number of eyes with satisfactory healing was greater in the biomembrane group than in the bare sclera group (p=0.06). The number of perilimbic vessels was significantly higher in the biomembrane group than in the bare sclera group (p=0.0284). The infection rate was similar in both groups. CONCLUSION: Similar to prior studies, the biomembrane of natural latex seemed to benefit conjunctival reconstruction and neoangiogenesis. Depending on the results using human eyes, this material might be a new type of therapeutic resource for conjunctival replacement, especially in cases of desirable tissue revascularization.

1 In this study, we examined the effects of crude venom from the spider Parawixia bistriata on glutamate and GABA uptake into synaptosomes prepared from rat cerebral cortex. Addition of venom to cortical synaptosomes stimulated glutamate uptake and inhibited GABA uptake in a concentration-dependent manner. 2 The venom was fractionated using reverse-phase high-performance liquid chromatograph on a preparative column. The fraction that retained glutamate uptake-stimulating activity was further purified on a reverse-phase analytical column followed by ion-exchange chromatography. 3 The active fraction, referred to as PbT � 1.2.3, stimulated glutamate uptake in synaptosomes without changing the KM value, and did not affect GABA uptake. Additional experiments showed that the enhancement of glutamate uptake by PbT � 1.2.3 occurs when ionotropic glutamate receptors or voltage-gated sodium and calcium channels are completely inhibited or when GABA receptors and potassium channels are activated, indicating that the compound may have a direct action on the transporters.

In this article we describe an in vivo anticonvulsant effect from denatured crude venom and partially isolated fractions from two spiders: Parawixia bistriata and Scaptocosa raptoria. Intracerebroventricular injections of these venoms and fractions abolished rat convulsive tonic-clonic seizures induced by picrotoxin, bicuculline and pentylenetetrazole, and also, inhibited GABA uptake in synaptosomes of rat cerebral cortex. The venoms described in this work seems to be promising tools for the study of the GABAergic system, and may be a potential source for new anticonvulsant drugs.

Hyaluronic acid hydrogels (HAHs) have been used as a carrier of substances and factors in the repair of nervous tissue. Natural latex protein (Hevea brasiliensis, F1) has shown positive effects in treating various types of tissues, including peripheral nerves. This study evaluated the F1 associated with a HAH in a controlled crush injury (axonotmesis) of the sciatic nerve in Wistar rats. The samples were photomicrographed for morphometric and quantitative analyzes using ImageJ 1.47k software (NIH, Bethesda, MD). Morphological, quantitative (myelin area/nerve area ratio and capillary density) and morphometric (minimum nerve fiber diameter, G-Ratio) data revealed an improvement in the recovery of the sciatic nerve with the application of HAH and the combination of HAH and F1 after 4 and 8 weeks of nerve injury. The most efficacious results were observed with the combination of both substances, F1 and HAH, revealing the regenerative capacity of this new biomaterial, which was hardly tested on nerve t...

This study evaluated the effect of low-level laser therapy (LLLT; 15 J/cm2) and a latex protein (F1) on a crush injury of the sciatic (ischiadicus) nerve. Seventy-two rats (male, 250 g) were divided into 6 groups: CG, control; EG, exposed nerve; IG, injured nerve without treatment; LG, injured nerve with LLLT; HG, injured nerve with F1; and LHG, injured nerve with LLLT and F1. After 4 or 8 weeks, the animals were euthanized and samples of the sciatic nerve were collected for morphometric and high-resolution scanning electron microscopy (HRSEM) analysis. After 4 weeks, the morphometry revealed improvements in the treated animals, and the HG appeared to be the most similar to the CG; after 8 weeks, the injured groups showed improvements compared to the previous period, and the results of the treatment groups were more similar to one another. At HRSEM after 4 weeks, the treated groups were similar and showed improvement compared to the IG; after 8 weeks, the LHG and HG had the best results. In conclusion, the treatments resulted in improvement after the nerve injury, and this recovery was time-dependent. In addition, the use of the F1 resulted in the best morphometric and ultrastructural findings.

In this study we evaluated the characteristics of the tibialis anterior muscle after sciatic nerve crush and treatment with low-level laser therapy (LLLT) or the protein from natural latex (P1).We studied the following 6 groups of male Wistar rats: control (CG); exposed nerve (EG); injured nerve (IG); injured nerve with LLLT (LG); injured nerve with P1 (PG); and injured nerve with P1 and LLLT (LPG).After 4 weeks, muscle morphology showed improvement in the treated groups; after 8 weeks, the treated groups resembled controls, especially the PG. Morphometry revealed muscle fiber atrophy after nerve injury, with time-dependent recovery. Histochemical analysis revealed increased intermediate fiber area. The PG was more similar to controls with NADH staining, whereas the LPG more closely resembled controls with SDH staining.Treatment using only P1 proved most efficient, revealing a negative interaction between P1 and LLLT.

A isquemia cerebral é fenômeno eventualmente observado durante procedimentos neurocirúrgicos e em patologias clínicas resultando em déficits neurológicos incapacitantes ou mesmo na morte. Por tratar-se de problema grave e de difícil solução, vários estudos têm sido efetuados com o objetivo de elucidar os mecanismos do fenômeno isquêmico no sistema nervoso central (SNC) e abolir ou diminuir seus efeitos através das drogas que protegem os neurônios (neuroprotetoras). Vários neurotransmissores estão envolvidos na isquemia e entre eles o glutamato destaca-se pela sua maior concentração no SNC. O objetivo deste estudo foi avaliar a isquemia cerebral focal em ratos através da dosagem do glutamato e dos achados morfológicos em uma evolução temporal e demonstrar uma possível ação neuroprotetora do cetoprofeno. Foram utilizados 36 ratos Wistar, subdivididos em 4 grupos: um grupo controle e outro sham; e outros dois em que os animais foram submetidos a isquemia pela oclusão seletiva da artéria cerebral média por um fio obstrutor durante 15, 30 e 45 minutos. Os animais de um destes grupos foram tratados com cetoprofeno 15 minutos antes da isquemia. A isquemia foi avaliada através de estudo histopatológico e da dosagem do glutamato extracelular in vitro. A análise morfológica mostrou não haver diferenças entre os animais normais e do grupo sham. Nos animais submetidos a isquemia, as alterações apareceram aos 30 minutos e acentuaram-se aos 45. Os principais achados foram edema intersticial, desorganização cromatínica, vacuolização e desintegração nuclear. Os animais tratados com cetoprofeno apresentaram alterações semelhantes, porém menos intensas. Reduções nas dosagens in vitro do glutamato extracelular no córtex parietal dos animais submetidos a isquemia iniciaram-se a partir dos 30 minutos e acentuaram-se aos 45 e foram semelhantes nos animais com ou sem tratamento com cetoprofeno, indicando que esta droga parece não interferir com o metabolismo do glutamato na sinapse. Os achados histopatológicos no córtex parietal dos animais submetidos a isquemia , tratados ou não previamente com cetoprofeno, sugerem que esta droga tem um efeito neuroprotetor.Cerebral ischaemia is eventualy observed during neurosurgical procedures and in several clinical entities that may cause severe neurological deficits and even death. Because it is a severe and complex problem, several studies have been done aiming to elucidate the mechanisms of the ischemic phenomenon and aiming to abolish or to diminish its effects, using drugs that protect the neurons from ischaemia-induced damage. Several neurotransmitters play a role in cerebral ischaemia with emphasis to glutamate by its high concentration in the central nervous system. The purpose of this study was to evaluate the effect of focal cerebral ischaemia in the rat through the dosage of the glutamate and morphological findings, and to evaluate a possible protective effect of the ketoprofen to ischemic neurons. Thirty-six rats Wistar were divided into four groups. The first was a control group, the second a sham group and the animals of the third and fourth groups were submitted to induced cerebral ischaemia through selective obstruction of the midlle cerebral artery during 15, 30 and 45 minutes. Animals of the fourth group were previously treated with ketoprofen 15 minutes before the ischaemia. The ischaemia was evaluated through the histopathological examination and through dosage of the extracellular glutamate in vitro. The histopathological examination showed that there was no difference between the animals of the control and of the sham groups. In the animals submitted to ischemia histopathological alterations appeared at 30 minutes and become more intense at 45 minutes of ischaemia. The main findings were interstitial edema, chromatinic disorganization, vacuolization and nuclear desintegration. The animals treated with ketoprofen showed similar alterations, but they were less intense. Decrease in the dosage of glutamate in the parietal cortex of the animals submitted to ischaemia started at 30 minutes and became more intense at 45 minutes of ischaemia and was similar for animals previously treated or not with ketoprofen, indicating that this drug seems not to interfere with the metabolism of the glutamate at the synapses. The morphological findings in the parietal cortex of the animals submitted to ischaemia, previously treated or not with ketoprofen, suggest that this drug has a neuroprotective effect.

FUNDAMENTO: São numerosas as vantagens de se fechar a cavidade pericárdica após as operações cardíacas e pneumonectomias intrapericárdicas. OBJETIVO: Estudar o comportamento da membrana de látex natural como substituto parcial do pericárdio. MATERIAL E MÉTODOS: Em 16 cães, divididos em 3 grupos, ressecou-se um retalho elíptico da porção ântero-lateral esquerda do pericárdio (7 cm x 5 cm): Grupo A (n=4) - o retalho removido foi reimplantado imediatamente; Grupo B (n=8) - o retalho foi substituído por outro, de látex natural, com área equivalente e espessura de 0,3 mm; Grupo C (n=4) - retalho de látex de 0,7 mm de espessura. Em todos os animais, fixou-se o retalho com sutura contínua de fio de polipropileno 5-0 ou 6-0. No último grupo, foram dados 4 pontos adicionais, em U, ancorados em barras de dacron. Realizaram-se eletrocardiogramas (ECG) e leucogramas no pré e no pós-operatório, bem como estudo macro e microscópio post-mortem. RESULTADOS: Grupo A - auto-enxerto íntegro, macro e microscopicamente, fortemente aderido ao pulmão e frouxamente aderido ao epicárdio; Grupo B - deiscência parcial da sutura em 1 e total em 2 animais. A membrana de látex não aderiu nem ao pulmão nem ao epicárdio. Em 3 (37,5%) animais houve regeneração total e distinta do pericárdio subjacente ao látex, microscopicamente idêntico ao pericárdio nativo. Grupo C - suturas íntegras. Em 3 (75%) animais observou-se regeneração pericárdica total e distinta. Nos demais animais dos Grupos B e C, e epicárdio subjacente ao látex apresentava-se ligeiramente espessado, permitindo visibilizar os vasos coronários com facilidade. Microscopicamente, identificaram-se algumas áreas com denso infiltrado linfo-plasmocitário, proliferação fibroblástica e vascular; em outras, os fibroblastos circunscreviam fendas com revestimento mesotelial, indicativas de regeneração irregular do pericárdio. Não se observou infecção local nem alteração do leucograma e ao ECG, apenas inversão da onda T, em todos os grupos. CONCLUSÃO: A membrana de látex natural mostrou-se adequada para a substituição parcial do pericárdio de cães, em observação de até 345 dias, propiciando a regeneração do pericárdio nativo.BACKGROUND: There are numerous advantages in closing the pericardial cavity after heart surgery and intrapericardial pneumonectomies. OBJECTIVE: To study the behavior of a natural latex membrane as a partial pericardium substitute. MATERIAL AND METHODS: Sixteen dogs divided into 3 groups were submitted to resection of an elliptical flap of the left anterolateral portion of the pericardium (7 x 5 cm): Group A (n=4) - the removed flap was immediately reimplanted; Group B (n=8) - the flap was replaced with a 0.3 mm thick natural latex membrane of equivalent area; Group C (n=4) - the latex flap was 0.7 mm thick. In all animals the flap was fixed with continuous 5-0 or 6-0 polypropylene sutures. In group C, 4 additional U-shaped stitches anchored with dacron pledgets were applied. All animals were submitted to ECG and to white cell counts during the preoperative period as well as to macro- and microscopic post mortem study. RESULTS: Group A - the self-graft was macro and microscopically intact, strongly adhering to the lung and loosely adhering to the epicardium; Group B - total suture dehiscence occurred in 1 animal and partial dehiscence in 2. The latex membrane did not adhere to the lung or to the epicardium. In 3 animals (37.5%) there was full regeneration of the pericardium underlying the natural latex, microscopically identical to the native pericardium. Group C - the sutures were intact and total pericardium regeneration was observed in 3 animals (75%). In the remaining animals of Groups B and C, the epicardium underlying the latex was slightly thickened, permitting easy visualization of the coronary vessels. Some areas with dense lymphocytes and plasma cells infiltrates, fibroblast and vascular proliferation were microscopically identified in its thickness; in some other areas the fibroblasts surrounded slits with mesothelial lining indicating irregular pericardium regeneration. No local infection or changes in white cell counts were observed and the ECG only showed T wave inversion in all groups. CONCLUSION: The natural latex membrane proved to be adequate for partial pericardium replacement in dogs kept under observation for up to 345 days, favoring regeneration of the native pericardium.

Glutamate (L-glu) is the most important excitatory neurotransmitter in the mammalian central nervous system. Its action is terminated by transporters located in the plasma membrane of neurons and glial cells, which have a critical role in preventing glutamate excitotoxicity under normal conditions. The neurotransmitter gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the mammalian central nervous system. Venoms of solitary wasps and orb-spiders are composed of large proteins, medium-size peptides, polyamine amides (PAs), and other neuroactive components that are highly selective to nervous tissues. The abnormal operation of uptake systems is involved in several failures. Several studies indicate alterations in extracellular GABA and glutamate concentrations in epilepsy conditions that may relate to transporter functions. The effects of the crude and boiled venom of the social wasp Agelaia vicina, "cassununga," on GABA and L-glu uptake in rat cerebral cortex synaptosomes are related. The venom uncompetitively inhibited high- and low-affinity GABA uptake by 91.2% and by 76%, respectively. This kind of inhibition was also found to affect high- (99.6%) and low-affinity (90%) uptake of L-glu. These results suggest that the effects observed in these experiments indicate the venom of A. vicina to be a useful tool to further characterize GABA- and L-glu-uptake systems.

FUNDAMENTO: São numerosas as vantagens de se fechar a cavidade pericárdica após as operações cardíacas e pneumonectomias intrapericárdicas. OBJETIVO: Estudar o comportamento da membrana de látex natural como substituto parcial do pericárdio. MATERIAL E MÉTODOS: Em 16 cães, divididos em 3 grupos, ressecou-se um retalho elíptico da porção ântero-lateral esquerda do pericárdio (7 cm x 5 cm): Grupo A (n=4) - o retalho removido foi reimplantado imediatamente; Grupo B (n=8) - o retalho foi substituído por outro, de látex natural, com área equivalente e espessura de 0,3 mm; Grupo C (n=4) - retalho de látex de 0,7 mm de espessura. Em todos os animais, fixou-se o retalho com sutura contínua de fio de polipropileno 5-0 ou 6-0. No último grupo, foram dados 4 pontos adicionais, em U, ancorados em barras de dacron. Realizaram-se eletrocardiogramas (ECG) e leucogramas no pré e no pós-operatório, bem como estudo macro e microscópio post-mortem. RESULTADOS: Grupo A - auto-enxerto íntegro, macro e microscopicamente, fortemente aderido ao pulmão e frouxamente aderido ao epicárdio; Grupo B - deiscência parcial da sutura em 1 e total em 2 animais. A membrana de látex não aderiu nem ao pulmão nem ao epicárdio. Em 3 (37,5%) animais houve regeneração total e distinta do pericárdio subjacente ao látex, microscopicamente idêntico ao pericárdio nativo. Grupo C - suturas íntegras. Em 3 (75%) animais observou-se regeneração pericárdica total e distinta. Nos demais animais dos Grupos B e C, e epicárdio subjacente ao látex apresentava-se ligeiramente espessado, permitindo visibilizar os vasos coronários com facilidade. Microscopicamente, identificaram-se algumas áreas com denso infiltrado linfo-plasmocitário, proliferação fibroblástica e vascular; em outras, os fibroblastos circunscreviam fendas com revestimento mesotelial, indicativas de regeneração irregular do pericárdio. Não se observou infecção local nem alteração do leucograma e ao ECG, apenas inversão da onda T, em todos os grupos. CONCLUSÃO: A membrana de látex natural mostrou-se adequada para a substituição parcial do pericárdio de cães, em observação de até 345 dias, propiciando a regeneração do pericárdio nativo. BACKGROUND: There are numerous advantages in closing the pericardial cavity after heart surgery and intrapericardial pneumonectomies. OBJECTIVE: To study the behavior of a natural latex membrane as a partial pericardium substitute. MATERIAL AND METHODS: Sixteen dogs divided into 3 groups were submitted to resection of an elliptical flap of the left anterolateral portion of the pericardium (7 x 5 cm): Group A (n=4) - the removed flap was immediately reimplanted; Group B (n=8) - the flap was replaced with a 0.3 mm thick natural latex membrane of equivalent area; Group C (n=4) - the latex flap was 0.7 mm thick. In all animals the flap was fixed with continuous 5-0 or 6-0 polypropylene sutures. In group C, 4 additional U-shaped stitches anchored with dacron pledgets were applied. All animals were submitted to ECG and to white cell counts during the preoperative period as well as to macro- and microscopic post mortem study. RESULTS: Group A - the self-graft was macro and microscopically intact, strongly adhering to the lung and loosely adhering to the epicardium; Group B - total suture dehiscence occurred in 1 animal and partial dehiscence in 2. The latex membrane did not adhere to the lung or to the epicardium. In 3 animals (37.5%) there was full regeneration of the pericardium underlying the natural latex, microscopically identical to the native pericardium. Group C - the sutures were intact and total pericardium regeneration was observed in 3 animals (75%). In the remaining animals of Groups B and C, the epicardium underlying the latex was slightly thickened, permitting easy visualization of the coronary vessels. Some areas with dense lymphocytes and plasma cells infiltrates, fibroblast and vascular proliferation were microscopically identified in its thickness; in some other areas the fibroblasts surrounded slits with mesothelial lining indicating irregular pericardium regeneration. No local infection or changes in white cell counts were observed and the ECG only showed T wave inversion in all groups. CONCLUSION: The natural latex membrane proved to be adequate for partial pericardium replacement in dogs kept under observation for up to 345 days, favoring regeneration of the native pericardium.