Plants used as antidiabetics in popular medicine in Rio Grande do Sul, southern Brazil.

ETHNOPHARMACOLOGICAL RELEVANCE: Plants are widely as antidiabetics. The study of these plants is essential because many of them may have undesirable effects, such as acute or chronic toxicity; or their use may even delay or discourage the adoption of the proper and effective treatment. MATERIALS AND METHODS: The present study surveyed the plant species that are popularly used to treat diabetes mellitus in the state of Rio Grande do Sul in southern Brazil. Sixteen ethnobotanical surveys performed in the state were consulted, and the species used to treat diabetes were listed. For species cited in at least two of the studies, scientific data related to antidiabetic activity were searched in the ISI Knowledge database. The scientific binomial of each species was used as keywords, and data found in review papers were also included. RESULTS: A total of 81 species in 42 families were mentioned; the most important families were Asteraceae and Myrtaceae. Twenty eight species were cited at least twice as being used to treat diabetes in the state. For 11 of these, no scientific data regarding antidiabetic activity could be located. The species most frequently mentioned for use with diabetes were Syzygium cumini (Myrtaceae) and Bauhinia forficata (Fabaceae), in 12 studies each, followed by Sphagneticola trilobata (Asteraceae), in six studies; and Baccharis trimera (Asteraceae), Bidens pilosa (Asteraceae), Cynara scolymus (Asteraceae), and Leandra australis (Melastomataceae) in four studies each. Bauhinia forficata and Syzygium cumini have been studied in more detail for antidiabetic activity. CONCLUSIONS: A considerable number of plant species are traditionally used for the treatment of diabetes melitus in the Rio Grande do Sul State. The majority of those plants that have been studied for antidiabetic activity showed promising results, mainly for Bauhinia forficata and Syzygium cumini. However, for most of the plants mentioned, the studies are not sufficient to guarantee the efficacy and safety in the use of these plants in the treatment against diabetes.

Plantas usadas como medicinais no município de Ipê, RS, Brasil / Medicinal plants used by people from Ipê, Rio Grande do Sul State, Brazil

Foi realizado um levantamento das plantas medicinais utilizadas pela população do município de Ipê, RS, Brasil, visando uma investigação da medicina tradicional, melhoramento e racionalização das práticas medicinais populares, em um projeto envolvendo voluntários da comunidade. Foram entrevistadas 114 pessoas, que mencionaram 252 plantas, das quais foram identificadas 105 espécies, agrupadas em 48 famílias. As plantas coletadas e identificadas foram analisadas, de acordo com dados químicos, farmacológicos e/ou toxicológicos encontrados na literatura científica. Os resultados obtidos mostram que a população deste município faz uso de plantas que, ainda, não foram alvo de pesquisa química, farmacológica e/ou toxicológica e, também, que a mesma faz uso de onze espécies, que são reconhecidamente, responsáveis por efeitos colaterais indesejados, ou ainda, que são tóxicas.

Medicinal plants used by Brazilian people from Ipê city, in Rio Grande do Sul State, were the subject of a survey realized in order to investigate the traditional medicine in this comunity. The project involved 114 individuals who cited 252 medicinal plants. From these plants, 105 species were identified and classified into 48 botanical families. Assessments of all known plants were based on chemical, pharmacological and toxicological data searched in scientific literature. The results show that many of these plants were not yet chemically and/or pharmacologically and/or toxicologically studied, and eleven of them were related to be responsible for side effects or even toxicity in the consulted scientific literature.

Differential regulation of tyrosine phosphorylation in tumor cells by contortrostatin, a homodimeric disintegrin, and monomeric disintegrins echistatin and flavoridin.

The homodimeric disintegrin contortrostatin was compared directly to the monomeric disintegrins echistatin and flavoridin for the ability to affect protein tyrosine phosphorylation in tumor cells. It was observed that contortrostatin had a dramatic effect on the tyrosine phosphorylation status of several proteins in T24 human bladder cancer cells, including robust induction of phosphorylation of proteins in the range of 120-140 kDa. Echistatin alone had no effect on tyrosine phosphorylation in T24 cells, but dose-dependently inhibits the effects of contortrostatin when both are added simultaneously. Among the proteins that undergo tyrosine phosphorylation in response to contortrostatin treatment is CAS, a 130 kDa adapter protein involved in integrin signaling. Flavoridin alone was found to have no effect on CAS, but can completely block contortrostatin-induced phosphorylation of this protein in MDA-MB-435 cells. These observations strongly suggest that the homodimeric structure of contortrostatin functionally distinguishes it from other monomeric members of the disintegrin family.

Contortrostatin, a homodimeric disintegrin, actively disrupts focal adhesion and cytoskeletal structure and inhibits cell motility through a novel mechanism.

Integrins play a major role in the regulation of cell motility. They physically link the extracellular environment to the cytoskeleton and participate in large protein complexes known as focal adhesions. In this report, it is demonstrated that treatment of tumor cells with the homodimeric disintegrin contortrostatin induces integrin-mediated tyrosine phosphorylation events and causes severe disruptions in the actin cytoskeleton and disassembly of focal adhesion structures without affecting cellular adhesion to a reconstituted basement membrane. Included in this disruption is the tyrosine phosphorylation and altered subcellular localization of FAK. Through use of transfected 293 cells with specific integrin expression profiles and anti-alphavbeta3 mAbs, we demonstrate that these events are mediated exclusively by the alphavbeta3 integrin and are likely the result of contortrostatin-mediated crosslinking of this receptor at the cell surface, since monovalent disintegrins, flavoridin or echistatin do not induce such effects. Further, it is shown that contortrostatin potently inhibits motility in cells expressing the alphavbeta33 integrin. The results of this study describe a novel integrin-mediated mechanism by which cell motility can be inhibited and suggest an alternative approach to therapeutic intervention for cancer invasion and metastasis.

Contortrostatin, a snake venom disintegrin, induces alphavbeta3-mediated tyrosine phosphorylation of CAS and FAK in tumor cells.

Contortrostatin is a homodimeric disintegrin that inhibits platelet aggregation and cell adhesion to extracellular matrix proteins by blocking integrins. The effect of contortrostatin on integrin-mediated signaling in tumor cells was investigated by studying tyrosine phosphorylation events and activation of specific signaling molecules. We found that at concentrations as low as 1 nM, soluble contortrostatin activates integrin signals leading to increased tyrosine phosphorylation of FAK and CAS, and that these signals are abolished by inhibiting Src family kinases. Using transfected 293 cells expressing specific integrins, it was determined that contortrostatin-generated signals are mediated exclusively by the alphavbeta3 integrin. This observation was extended by showing that cells lacking alphavbeta3, but expressing alphavbeta5 and alpha5beta1, do not respond in this way to contortrostatin treatment. In cells expressing alphavbeta3, blocking contortrostatin binding with antibodies against alphavbeta3 completely abrogates contortrostatin signals. Monovalent disintegrins echistatin and flavoridin were incapable of affecting tyrosine phosphorylation alone, but when added simultaneously with contortrostatin, completely inhibited contortrostatin-initiated signals. We propose that the homodimeric nature of contortrostatin imparts the ability to crosslink alphavbeta3 integrins, causing Src activation and hyperphosphorylation of FAK and CAS. This activity may represent a novel mechanism by which tumor cell motility can be inhibited.

Contortrostatin activates ERK2 and tyrosine phosphorylation events via distinct pathways.

We report that cells adhering to contortrostatin show transient increases in activation of Extracellular signal Regulated Kinase 2 (ERK2). The kinetics and degree of activation are similar to cells adhering to fibronectin or vitronectin. We have recently shown that contortrostatin induces tyrosine phosphorylation in tumor cells. Contortrostatin is shown here to stimulate activation of ERK2 in suspended cells, but this activation follows a different dose-response pattern than contortrostatin-induced tyrosine phosphorylation. Since contortrostatin induces tyrosine phosphorylation via alphavbeta3, we explored the effects of an alphavbeta3-blocking antibody, 7E3, on contortrostatin-stimulated ERK2 activation. While 7E3 completely blocks the effect of contortrostatin on tyrosine phosphorylation, this antibody had no effect on activation of ERK2. In cells lacking expression of alphavbeta3, tyrosine phosphorylation was unaffected by contortrostatin treatment, but ERK2 was activated. This is strong evidence that contortrostatin is regulating tyrosine phosphorylation events and ERK2 activation via separate pathways and through different integrin receptors.

Molecular cloning and functional expression of contortrostatin, a homodimeric disintegrin from southern copperhead snake venom.

Contortrostatin is a unique dimeric disintegrin isolated from southern copperhead snake venom. Through antagonism of integrins alphaIIbbeta3, alpha5beta1, alphavbeta3, and alphavbeta5, contortrostatin inhibits platelet aggregation and disrupts cancer cell adhesion and invasion. We cloned cDNA from a library made from the venom gland cells of Agkistrodon contortrix contortrix using polymerase chain reaction. We found that the contortrostatin gene is part of a precursor composed of proprotein, metalloproteinase, and disintegrin domains. The precursor cDNA is 2027 bp with a 1449-bp open reading frame. The disintegrin domain is 195 bp encoding 65 amino acids. Like other members of the disintegrin family, each subunit of contortrostatin has an RGD site, and the cysteine alignment is conserved. The disintegrin domain of the cDNA has been expressed in a eukaryotic expression system as a homodimeric fusion protein with an immunoglobulin. The recombinant protein is recognized by an antiserum against native contortrostatin in Western blot. Both the native and recombinant proteins bind to integrins alphavbeta3 and alphavbeta5. Like native contortrostatin, the recombinant fusion protein inhibits platelet aggregation, blocks cancer cell adhesion to fibronectin and vitronectin, and prevents invasion of cancer cells through a Matrigel barrier. The success of functional expression not only validates the cDNA cloning of this disintegrin, but also provides adequate material for functional studies of contortrostatin.

Contortrostatin, a homodimeric disintegrin, binds to integrin alphavbeta5.

Contortrostatin is a homodimeric disintegrin from snake venom. We have shown that contortrostatin binds to integrins alphaIIbbeta3, alpha5beta1, and alphavbeta3. We now use several criteria to demonstrate the binding of contortrostatin to alphavbeta5. First, incubation of T24 cells, which express alphavbeta3 and alphavbeta5, with antibody against alphavbeta3 failed to completely inhibit adhesion of cells to vitronectin. However, pretreatment of the cells with contortrostatin or the combination of antibodies against alphavbeta3 and alphavbeta5 completely blocked adhesion to vitronectin. By contrast, either anti-alphavbeta5 alone or contortrostatin blocked adhesion of an alphavbeta3-negative T24 subline. Second, contortrostatin as well as anti-alphavbeta5 inhibits invasion of OVCAR-5, which express only alphavbeta5. Third, contortrostatin binds to purified alphavbeta5 in a saturable manner. Finally, radioligand binding assays yielded a K(d) value of 24 nM for [(125)I]contortrostatin binding to alphavbeta5. This investigation identifies alphavbeta5 as a binding site for contortrostatin. Blockage of alphavbeta5 by contortrostatin inhibits alphavbeta5-mediated adhesion and invasion.

The role of arthroscopy in the treatment of lunotriquetral ligament injuries.

The treatment of lunotriquetral interosseous (LTIO) ligament injuries is controversial. It is part of a spectrum of injuries involving the ulnar side of the wrist. Associated injuries may play a greater role in the outcome of treatment than does the LTIO injury. Arthroscopy is the most accurate diagnostic tool available for the evaluation of LTIO ligament injuries; it allows diagnosis and often treatment of associated injuries. This article describes the current approach to the diagnosis and treatment of LTIO ligament injuries and related use of arthroscopy.

Incorrect death certification. An invitation to obfuscation.

The physician responsible for signing a death certificate has an obligation to complete the medical part of the form, which includes entering the cause of death. In a review of 384 death certificates signed by house staff and attending physicians at a university hospital over a one-year period, 59% contained errors in cause-of-death entries. The most common error was listing mechanisms rather than causes of death. Since death certificates are used in calculating basic mortality statistics, such a large percentage of error can lead to considerable statistical misinformation.