Binding of the volatile general anesthetics halothane and isofluorane to a mammalian beta barrel protein

A molecular understanding of volatile anesthetic mechanisms of action will require structural descriptions of anesthetic-protein complexes. Porcine odorant binding protein is a 157 residue member of the lipocalin family that features a large beta-barrel internal cavity (515 +/- 30 angstroms(3)) lined predominantly by aromatic and aliphatic residues. Halothane binding to the beta-barrel cavity was determined using fluorescence quenching of Trp16, and a competitive binding assay with 1-aminoanthracene. In addition, the binding of halothane and isoflurane were characterized thermodynamically using isothermal titration calorimetry. Hydrogen exchange was used to evaluate the effects of bound halothane and isoflurane on global protein dynamics. Halothane bound to the cavity in the beta-barrel of porcine odorant binding protein with dissociation constants of 0.46 +/- 0.10 mM and 0.43 +/- 0.12 mM determined using fluorescence quenching and competitive binding with 1-aminoanthracene, respectively. Isothermal titration calorimetry revealed that halothane and isoflurane bound with K(d) values of 80 +/- 10 microM and 100 +/- 10 microM, respectively. Halothane and isoflurane binding resulted in an overall stabilization of the folded conformation of the protein by -0.9 +/- 0.1 kcal.mol(-1). In addition to indicating specific binding to the native protein conformation, such stabilization may represent a fundamental mechanism whereby anesthetics reversibly alter protein function. Because porcine odorant binding protein has been successfully analyzed by X-ray diffraction to 2.25 angstroms resolution [1], this represents an attractive system for atomic-level structural studies in the presence of bound anesthetic. Such studies will provide much needed insight into how volatile anesthetics interact with biological macromolecules

Ecological analogies between estuarine bottom trawl fish assemblages from Patos Lagoon, Rio Grande do Sul, Brazil and York River, Virginia, USA

The structure of estuarine fish assemblages at temperate latitudes in Patos Lagoon (32°05’S, 52°04’W), Rio Grande do Sul, Brazil and York River (37°17’N, 76°33’W), Virginia, USA was compared using mid and late 1970’s data from bottom trawl collection to investigate whether geographically isolated fish assemblages have similar ecological structure given similar latitudinal positions on the warm-temperate southwestern and northwestern Atlantic regions, respectively. Since estuarine species often exhibit an ontogenetic shift in habitat requirements or preferences we examined Capture per Unity of Effort by size class (CPUE-SC) and split species into “size ecological taxa” (SET) for analysis. The use of CPUE-SC also allowed the abundance of a SET to be computed by summing the mean CPUE of each size class within that SET and use this information to follows SET’s temporal and or spatial abundance. A total of 65 and 63 species was collected during a year of bottom trawling in the Patos Lagoon and York River estuaries, respectively. In both localities the strongest modal size class was < 80 mm TL, and several abundant species were smaller than 100 mm TL. The size between 80 and 100 TL effectively separated several species into discrete SET’s in both systems. Those SET’s could have different ecological preferences, temporal and spatial distributions and so identified as different "ecological taxa". In warm months, when predation by large fish is most likely, the abundance of fish between 80 and 100 mm TL in "bottom trawl" demersal fish assemblages was low in both systems. Only the sea catfishes, in Patos Lagoon, protected by strong dorsal and pectoral spines, and the Hogchoker, in the York River, protected by burrowing in the bottom substrate, peak in abundance at this size class. The seasonal pattern of estuarine use was similar between localities and did not differ from other warm-temperate estuarine fish assemblages.A estrutura das assembléias de peixes de dois estuários temperados – Lagoa dos Patos, Brasil e York River, Estados Unidos – foi comparada usando dados de rede de arrasto de fundo, da década de 70, em ambas as regiões. O objetivo do trabalho foi o de investigar se a fauna de peixes de dois estuários, localizados em duas regiões temperadas-quente do Atlântico, embora isolados geograficamente (Hemisfério Sul e Norte), apresentavam a mesma estrutura ecológica. Considerando a ontogenia das espécies utilizou-se o método da Captura por Unidade de Esforço por Classe de Tamanho (CPUE-CT) para separar as espécies dominantes em Unidades Ecológicas de Tamanho (UET). O uso da técnica do CPUE-CT permite que a abundância das UET seja computada através da soma da abundância de cada uma das classes de tamanho que incorporam estas UET e desta forma usar o CPUE das UET para estudar sua variação temporal ou espacial de abundância. Após um ano de coleta mensal foram observadas 65 espécies nas coletas de arrasto de fundo na Laguna dos Patos e 63 espécies no York River. Em ambas as localidades as maiores modas de tamanho foram menores que 80 mm de comprimento total (CT), e diversas espécies ocorrem em tamanho menor do que 100 mm CT. Entre 80 e 100 mm CT foi possível separar efetivamente diversas espécies em UET. Diferentes UET de uma mesma espécie ocorrem em diferentes habitats e em épocas distintas, podendo assim ser classificadas como “Taxas Ecológicos” distintos de uma mesma espécie. Nos meses quentes, quando ocorre a maior pressão de predação dos grandes peixes piscívoros, a abundância dos peixes com tamanhos entre 80 e 100 mm CT nas coletas de arrasto de fundo é baixa em ambos os sistemas. Somente dois grupos de peixes são abundantes neste tamanho, os bagres marinhos da Laguna dos Patos que estão protegidos da predação pelos espinhos dorsais e peitorais, e o linguado-Zebra no York River, que se enterra no substrato evitando a predação

The mechanisms of coronary restenosis: insights from experimental models

Since its introduction into clinical practice, more than 20 years ago, percutaneous transluminal coronary angioplasty (PTCA) has proven to be an effective, minimally invasive alternative to coronary artery bypass grafting (CABG). During this time there have been great improvements in the design of balloon catheters, operative procedures and adjuvant drug therapy, and this has resulted in low rates of primary failure and short-term complications. However, the potential benefits of angioplasty are diminished by the high rate of recurrent disease. Up to 40% of patients undergoing angioplasty develop clinically significant restenosis within a year of the procedure. Although the deployment of endovascular stents at the time of angioplasty improves the short-term outcome, ‘in-stent’ stenosis remains an enduring problem. In order to gain an insight into the mechanisms of restenosis, several experimental models of angioplasty have been developed. These have been used together with the tools provided by recent advances in molecular biology and catheter design to investigate restenosis in detail. It is now possible to deliver highly specific molecular antagonists, such as antisense gene sequences, to the site of injury. The knowledge provided by these studies may ultimately lead to novel forms of intervention. The present review is a synopsis of our current understanding of the pathological mechanisms of restenosis