Active site heterogeneity in dimethyl sulfoxide reductase from Rhodobacter capsulatus revealed by raman spectroscopy

Raman spectroscopy has been used to investigate the structure of the molybdenum cofactor in DMSO reductase from Rhodobacter capsulatus. Three oxidized forms of the enzyme, designated 'redox cycled', 'as prepared', and DMSORmodD, have been studied using 752 nm laser excitation. In addition, two reduced forms of DMSO reductase, prepared either anaerobically using DMS or using dithionite, have been characterized. The 'redox cycled' form has a single band in the Mo=O stretching region at 865 cm(-1) consistent with other studies. This oxo ligand is found to be exchangeable directly with (DMSO)-O-18 or by redox cycling. Furthermore, deuteration experiments demonstrate that the oxo ligand in the oxidized enzyme has some hydroxo character, which is ascribed to a hydrogen bonding interaction with Trp 116. There is also evidence from the labeling studies for a modified dithiolene sulfur atom, which could be present as a sulfoxide. In addition to the 865 cm(-1) band, an extra band at 818 cm(-1) is observed in the Mo=O stretching region of the 'as prepared' enzyme which is not present in the 'redox cycled' enzyme. Based on the spectra of unlabeled and labeled DMS reduced enzyme, the band at 818 cm(-1) is assigned to the S=O stretch of a coordinated DMSO molecule. The DMSORmodD form, identified by its characteristic Raman spectrum, is also present in the 'as prepared' enzyme preparation but not after redox cycling. The complex mixture of forms identified in the 'as prepared' enzyme reveals a substantial degree of active site heterogeneity in DMSO reductase.

Expression of the hepatitis C virus structural proteins in mammalian cells induces morphology similar to that in natural infection

Like many positive-strand RNA viruses, replication of the hepatitis C virus (HCV) is associated with cytoplasmic membrane rearrangements. However, it is unclear which HCV Proteins induce these ultrastructural features. This work examined the morphological changes induced by expression of the HCV structural proteins, core, E1 and E2, expressed from a Semliki Forest Virus (SFV) recombinant RNA replicon. Electron microscopy of cells expressing these proteins showed cytoplasmic vacuoles containing membranous and electron-dense material that were distinct from the type I cytoplasmic vacuoles induced during SFV replicon replication. Immunogold labelling showed that the core and E2 proteins localized to the external and internal membranes of these vacuoles. At times were also associated with some of the internal amorphous material. Dual immunogold labelling with antibodies raised against the core protein and against an endoplasmic reticulum (ER)-resident protein (protein disulphide isomerase) showed that the HCV-induced vacuoles were associated with ER-labelled membranes. This report has identified an association between the HCV core and E2 proteins with induced cytoplasmic vacuoles which are morphologically similar to those observed in HCV-infected liver tissue, suggesting that the HCV structural proteins may be responsible for the induction of these vacuoles during HCV replication in vivo.

Development and evaluation of a species diagnostic polymerase chain reaction-restriction fragment-length polymorphism procedure for cryptic members of the Culex sitiens (Diptera : Culicidae) subgroup in Australia and the southwest Pacific

Members of the Culex sitiens subgroup are important vectors of arboviruses, including Japanese encephalitis virus, Murray Valley encephalitis virus and Ross River virus. Of the eight described species, Cx. annulirostris Skuse, Cx. sitiens Wiedemann, and Cx. palpalis Taylor appear to be the most abundant and widespread throughout northern Australia and Papua New Guinea (PNG). Recent investigations using allozymes have shown this subgroup to contain cryptic species that possess overlapping adult morphology. We report the development of a polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) procedure that reliably separates these three species. This procedure utilizes the sequence variation in the ribosomal DNA ITS1 and demonstrates species-specific PCR-RFLP profiles from both colony and field collected material. Assessment of the consistency of this procedure was undertaken on mosquitoes sampled from a wide geographic area including Australia, PNG, and the Solomon Islands. Overlapping adult morphology was observed for Cx. annulirostris and Cx. palpalis in both northern Queensland and PNG and for all three species at one site in northwest Queensland.

Microstructural evolution of the Serido Belt, NE-Brazil: the effect of two tectonic events on development of c-axis preferred orientation in quartz

The polyphase evolution of the Serido Belt (NE-Brazil) includes D, crust formation at 2.3-2.1 Ga, D-2 thrust tectonics at 1.9 Ga and crustal reworking by D-3 strike-slip shear zones at 600 Ma. Microstructural investigations within mylonites associated with D-2 and D-3 events were used to constrain the tectono-thermal evolution of the belt. D-2 shear zones commenced at deeper crustal levels and high amphibolite facies conditions (600-650 degreesC) through grain boundary migration, subgrain rotation and operation of quartz Q-prism slip. Continued shearing and exhumation of the terrain forced the re-equilibration of high-T fabrics and the switching of slip systems from (c)-prism to positive and negative (a)-rhombs. During D-3, enhancement of ductility by dissipation of heat that came from syn-D-3 granites developed wide belts of amphibolite facies mylonites. Continued shearing, uplift and cooling of the region induced D-3 shear zones to act in ductile-brittle regimes, marked by fracturing and development of thinner belts of greenschist facies mylonites. During this event, switching from (a)-prism to a basal slip indicates a thermal path from 600 to 350 degreesC. Therefore, microstructures and quartz c-axis fabrics in polydeformed rocks from the Serido Belt preserve the record of two major events, which includes contrasting deformation mechanisms and thermal paths. (C) 2003 Elsevier Ltd. All rights reserved.

The egalitarian leader: A comparison of leadership in Australia and New Zealand

This article compares leadership in Australia and New Zealand based on data collected as a part of the GLOBE (Global Leadership and Organizational Behavior effectiveness) 62-nation culture and leadership project. Exploratory and confirmatory factor analyses were used to demonstrate that etic (universal) dimensions of 'Charismatic' and 'Self-Protective' leadership are evident in both cultures, but that the dimensions have emic (local) culturally determined manifestations. These emic manifestations were stronger in New Zealand than in Australia. Leadership effectiveness incorporated the negative emic dimension of 'Bureaucratic' leadership (both countries), and the positive emic dimension of 'Egalitarian leadership' in Australia and 'Team leadership' in New Zealand. Both models of leadership nonetheless represent styles of leadership based on egalitarian principles.

Invasions changing marine biodiversity. Comment on J.M. Pandolfi & C.E. Lovelock Perspective in Ecology "Novelty Trumps Loss in Global Biodiversity" (18 April, 344, 266).

Although global biodiversity is declining, local ecosystems are not systematically losing diversity, but rather experiencing rapid turnover in species.

Solubilities of C‑Tetraalkylcalix[4]resorcinarenes in SCCO2: experimental measurements, characterization, and correlation

The solubilities of two C-tetraalkylcalix[4]resorcinarenes, namely C-tetramethylcalix[4]resorcinarene and C-tetrapentylcalix[4]resorcinarene, in supercritical carbon dioxide (SCCO2) were measured in a flow-type apparatus at a temperature range from (313.2 to 333.2) K and at pressures from (12.0 to 35.0) MPa. The C-tetraalkylcalix[4]resorcinarenes were synthesized applying our optimized procedure and fully characterized by means of gel permeation chromatography, infrared and nuclear magnetic resonance spectroscopy. The solubilities of the C-tetraalkylcalix[4]resorcinarenes in SCCO2 were determined by analysis of the extracts obtained by HPLC with ultraviolet (UV) detection methodology adapted by our team. Four semiempirical density-based models, and the SoaveRedlichKwong cubic equation of state (SRK CEoS) with classical mixing rules, were applied to correlate the solubility of the calix[4]resorcinarenes in the SC CO2. The physical properties required for the modeling were estimated and reported.

CLINICAL AND EPIDEMIOLOGICAL PROFILE OF ELDERLY PATIENTS WITH CHAGAS DISEASE FOLLOWED BETWEEN 2005-2013 BY PHARMACEUTICAL CARE SERVICE IN CEARÁ STATE, NORTHEASTERN BRAZIL

By controlling the transmission of Chagas disease, the challenge of providing assistance to millions of infected patients that reach old age arises. In this study, the socioeconomic, demographic and comorbidity records of all elderly chagasic patients followed at the Pharmaceutical Care Service of the Chagas Disease Research Laboratory were assessed. The information related to the clinical form of the disease was obtained from medical records provided by the Walter Cantídio University Hospital. The profile of the studied population was: women (50.5%); mean age of 67 years; retired (54.6%); married (51.6 %); high illiteracy rate (40.2%); and family income equal to the minimum wage (51.5%). The predominant clinical forms of Chagas disease were cardiac (65.3%) and indeterminate (14.7%). The main electrocardiographic changes were the right bundle branch block (41.0%), associated or not with the anterosuperior left bundle branch block (27.4%). The average number of comorbidities per patient was 2.23 ± 1.54, with systemic arterial hypertension being the main one found (67.0%). It was found that the elderly comprise a vulnerable group of patients that associate aging with cardiac and/or digestive disorders resulting from the evolution of Chagas disease and other comorbidities, which requires special attention from health services to ensure more appropriate medical and social care.

Surgical site infections in women and their association with clinical conditions

Introduction Surgical site infections (SSIs) can affect body tissues, cavities, or organs manipulated in surgery and constitute 14% to 16% of all infections. This study aimed to determine the incidence of SSIs in women following their discharge from a gynecology outpatient clinic, to survey different types of SSIs among women, and to verify the association of SSIs with comorbidities and clinical conditions. Methods Data were collected via analytical observation with a cross-sectional design, and the study was conducted in 1,026 women who underwent gynecological surgery in a teaching hospital in the municipality of Teresina, in the northeast Brazilian State of Piauí, from June 2011 to March 2013. Results The incidence of SSIs after discharge was 5.8% among the women in the outpatient clinic. The most prevalent surgery among the patients was hysterectomy, while the most prevalent type of SSI was superficial incisional. Comorbidities in women with SSIs included cancer, diabetes mellitus, and hypertension. Conclusions Surveillance of SSIs during the post-discharge period is critical for infection prevention and control. It is worth reflecting on the planning of surgical procedures for patients who have risk factors for the development of SSIs.

Occult HBV infection status among chronic hepatitis C and hemodialysis patients in Northeastern Egypt: regional and national overview

INTRODUCTION: Occult hepatitis B infection (OBI) is considered to be one of the major risks for patients suffering from end-stage renal disease (ESRD) on regular hemodialysis (HD) and patients with chronic hepatitis C virus (HCV) infection. This study compared the prevalence of OBI among these two high-risk groups in the Suez Canal region, Northeastern Egypt, to obtain a better national overview of the magnitude of OBI in this region. METHODS: Serum samples were collected from 165 HD patients and 210 chronic HCV-infected patients. Anti-HCV antibody, hepatitis B surface antigen (HBsAg), total hepatitis B core (anti-HBc) antibody, and hepatitis B surface antibody (anti-HBs) were detected by enzyme-linked immunosorbent assay (ELISA). HCV RNA was detected using a quantitative real-time RT-PCR assay, and HBV was detected using a nested PCR. RESULTS: All patients were negative for HBsAg. A total of 49.1% and 25.2% of the patients in the HD and HCV groups, respectively, were anti-HBc-positive. In addition, more anti-HBs-positive patients were detected in the HD group compared to the HCV group (52.1% and 11.4%, respectively). Three cases were positive for HBV DNA in the HD group, while eighteen positive cases were detected in the HCV group. Both study groups showed significant differences in serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) level as well as anti-HBc, anti-HBs and HBV-DNA positivity. CONCLUSIONS: OBI was more prevalent among chronic HCV patients than HD patients in the Suez Canal region, Egypt, with rates of 8.5% and 1.8%, respectively. However, more precise assessment of this infection requires regular patient follow-up using HBV DNA detection methods.

On project finance and the first road concession in Portugal

In broad sense, Project Financing1 as a mean of financing large scale infrastructural projects worldwide has had a steady growth in popularity for the last 20 years. This growth has been relatively unscathed from most economic cycles. However in the wake of the 2007 systemic Financial Crisis, Project Financing was also in trouble. The liquidity freeze and credit crunch that ensued affected all parties involved. Traditional Lenders, of this type of financial instrument, locked-in long-term contractual obligations, were severely hit with scarcity of funding compounded by rapidly increasing cost of funding. All the while, Banks were “rescued” by the concerted actions of Central Banks and other Multi-Lateral Agencies around the world but at the same time “stressed” by upcoming regulatory effort (Basel Committee). This impact resulted in specific changes to this type of long-term financing. Changes such as Commercial Banks’ increased risk aversion; pricing increase and maturities decrease of credit facilities; enforcement of Market Disruption Event clauses; partial responsibility for project risk by Multilateral Agencies; and adoption of utility-like availability payments in other industrial sectors such as transportation and even social infrastructure. To the extent possible, this report is then divided in three parts. First, it begins with a more instructional part, touching academic literature (theory) and giving the Banks perspective (practice), but mostly as an overview of Project Finance for awareness’ sake. The renowned Harvard Business School professor – Benjamin Esty, states2 that Project Finance is a “relatively unexplored territory for both empirical and theoretical research” which means that academic research efforts are lagging the practice of Project Finance. Second, the report presents a practical case regarding the first Road Concession in Portugal in 1998 ending with the lessons learned 10 years after Financial Close. Lastly, the report concludes with the analysis of the current trends and changes to the industry post Financial Crisis of the late 2000’s. To achieve this I’ll reference relevant papers, books on the subject, online articles and my own experience in the Project Finance Department at a major Portuguese Investment Bank. Regarding the latter, with the signing of a confidentiality agreement, I’m duly omitting sensitive and proprietary bank information.

Technical requirements for the use of C&DW recycled aggregates in municipal and rural roads and trenches

The use of construction and demolition waste (C&DW) in the construction industry is an important contribution to attain sustainability in the sector. The roads are among the civil engineering works which can use larger quantities of C&DW recycled aggregates. In Portugal, the limit values for the properties of C&DW recycled aggregates that can be used in the roads of Portuguese Road Network are defined by two Laboratório Nacional de Engenharia Civil (LNEC) technical specifications (TS), in accordance to Portuguese Decree-law no. 46/2008 of May 12th. Municipal and rural roads and trenches have specific characteristics that can enable the use of C&DW of lower quality than those required by existing LNEC TS, and even then ensuring an adequate performance. However, given the absence of specific regulation for those applications, the Portuguese Environment Agency requires compliance with the existing LNEC TS, which represents an obstacle to recycling a significant part of the C&DW, in particular at a local government level. This paper presents guidelines for the recycling of C&DW in municipal and rural roads and in trenches, which could be considered in a new forthcoming LNEC TS. In the preparation of the guidelines, the bibliography collected and analysed, the information gathered from the application of C&DW in a municipal and rural roads of a Portuguese municipality and in the roadways of a Portuguese resort, and the results of laboratory tests carried out on samples collected in the Portuguese municipality were taken into consideration.

apoptosis-regulator c-FLIP : functional role in urothelial carcinoma and autoimmunity and identification of novel CD95 DISC-interacting proteins

Magdeburg, Univ., Fak. für Naturwiss., Diss., 2013

C*-algebras nearly contained in type I algebras

"Vegeu el resum a l'inici del document del fitxer adjunt"

C-banding and FISH in chromosomes of the blow flies Chrysomya megacephala and Chrysomya putoria (Diptera, Calliphoridae)

The blow flies Chrysomya putoria and C. megacephala have 2n=12 chromosomes, five metacentric pairs of autosomes and an XX/XY sex chromosome pair. There are no substantial differences in the karyotype morphology of these two species, except for the X chromosome which is subtelocentric in C. megacephala and metacentric in C. putoria and is about 1.4 times longer in C. putoria. All autosomes were characterized by the presence of a C band in the pericentromeric region; C. putoria also has an interstitial band in pair III. The sex chromosomes of both species were heterochromatic, except for a small region at the end of the long arm of the X chromosome. Ribosomal genes were detected in meiotic chromosomes by FISH and in both species the NOR was located on the sex chromosomes. These results confirm that C. putoria was the species introduced into Brazil in 1970s, and not C. chloropyga as formerly described.