Carvedilol inhibition of lipid peroxidation. A new antioxidative mechanism

To define the molecular mechanism(s) of carvedilol inhibition of lipid peroxidation we have utilized model systems that allow us to study the different reactions involved in this complex process.

Carvedilol inhibits the peroxidation of sonicated phosphatidylcholine liposomes triggered by FeCl₂ addition whereas atenolol, pindolol and labetalol are ineffective. The inhibition proved not to be ascribable (a) to an effect on Fe²⁺ autoxidation and thus on the generation of oxygen derived radical initiators; (b) to the scavenging of the inorganic initiators O^·-₂ and ^·OH; (c) to an effect on the reductive cleavage of organic hydroperoxides by FeCl₂; (d) to the scavenging of organic initiators. The observations that (a) carvedilol effectiveness is inversely proportional to the concentration of FeCl₂ and lipid hydroperoxides in the assay; (b) the drug prevents the onset of lipid peroxidation stimulated by FeCl₃ addition and; (c) it can form a complex with Fe³⁺, suggest a molecular mechanism for carvedilol action. It may inhibit lipid peroxidation by binding the Fe³⁺ generated during the oxidation of Fe²⁺ by lipid hydroperoxides in the substrate. The lag time that carvedilol introduces in the peroxidative process would correspond to the time taken for carvedilol to be titrated by Fe³⁺; when the drug is consumed the Fe³⁺ accumulates to reach the critical parameter that stimulates peroxidation. According to this molecular mechanism the antioxidant potency of carvedilol can be ascribed to its ability to bind a species, Fe³⁺, that is a catalyst of the process and to its lipophilic nature that concentrates it in the membranes where Fe³⁺ is generated by a site specific mechanism.     

Colon tumour secretopeptidome: Insights into endogenous proteolytic cleavage events in the colon tumour microenvironment

The secretopeptidome comprises endogenous peptides derived from proteins secreted into the tumour microenvironment through classical and non-classical secretion. This study characterised the low-M_r (< 3 kDa) component of the human colon tumour (LIM1215, LIM1863) secretopeptidome, as a first step towards gaining insights into extracellular proteolytic cleavage events in the tumour microenvironment. Based on two biological replicates, this secretopeptidome isolation strategy utilised differential centrifugal ultrafiltration in combination with analytical RP-HPLC and nanoLC-MS/MS. Secreted peptides were identified using a combination of Mascot and post-processing analyses including MSPro re-scoring, extended feature sets and Percolator, resulting in 474 protein identifications from 1228 peptides (≤ 1% q-value, ≤ 5% PEP) — a 36% increase in peptide identifications when compared with conventional Mascot (homology ionscore thresholding). In both colon tumour models, 122 identified peptides were derived from 41 cell surface protein ectodomains, 23 peptides (12 proteins) from regulated intramembrane proteolysis (RIP), and 12 peptides (9 proteins) generated from intracellular domain proteolysis. Further analyses using the protease/substrate database MEROPS, (http://merops.sanger.ac.uk/), revealed 335 (71%) proteins classified as originating from classical/non-classical secretion, or the cell membrane. Of these, peptides were identified from 42 substrates in MEROPS with defined protease cleavage sites, while peptides generated from a further 205 substrates were fragmented by hitherto unknown proteases. A salient finding was the identification of peptides from 88 classical/non-classical secreted substrates in MEROPS, implicated in tumour progression and angiogenesis (FGFBP1, PLXDC2), cell–cell recognition and signalling (DDR1, GPA33), and tumour invasiveness and metastasis (MACC1, SMAGP); the nature of the proteases responsible for these proteolytic events is unknown. To confirm reproducibility of peptide fragment abundance in this study, we report the identification of a specific cleaved peptide fragment in the secretopeptidome from the colon-specific GPA33 antigen in 4/14 human CRC models. This improved secretopeptidome isolation and characterisation strategy has extended our understanding of endogenous peptides generated through proteolysis of classical/non-classical secreted proteins, extracellular proteolytic processing of cell surface membrane proteins, and peptides generated through RIP. The novel peptide cleavage site information in this study provides a useful first step in detailing proteolytic cleavage associated with tumourigenesis and the extracellular environment. This article is part of a Special Issue entitled: An Updated Secretome.     

A benign approach to the preparation of freshwater bryozoan statoblasts for scanning electron microscopy (SEM) imaging

Abstract

Several different species of freshwater Bryozoa, belonging to the genera Plumatella, Rumarcanella and Fredericella, were detected within the Northern Mallee Pipeline (NMP) system in Victoria, Australia, that required definitive identification. These organisms produce asexual buds called statoblasts, with valves composed of sclerotised chitin that bear minute micro-ornamentations of considerable taxonomical significance. Imaging and analysis of these distinctive micro-ornamentations using scanning electron microscopy (SEM) is often employed for species identification. Meticulous preparation of statoblast samples is therefore required that necessitates the removal of adhering debris, dehydration and drying—whilst mitigating specimen damage and distortion. This technical note describes an approach whereby each of these three steps have been individually designed to be as benign as possible, using mild detergent/sonication to remove debris, a gradual and gentle dehydration procedure using ethanol, and critical point drying. For the overall process, these methods are chosen to optimise control and to minimise the use of harsh and hazardous chemicals.     

Disaffected black and white youth: The role of unemployment duration and perceived job discrimination*

While unauthorized immigration has existed in the USA since the inception of immigration laws in the early twentieth century, ‘illegality’ did not become a central concern in mainstream debate until the late 1970s. Existing scholarship has developed two lines of argument to explain the salience of illegality: a state-centred approach that sees bureaucrats pushing forth the category, and a ‘bottom-up’ approach that emphasizes the grass-roots activism of restrictionist organizations effectively disguising their nativism by appealing to law and order. The data collected here builds on but complicates the state-centred explanation, and points away from the ‘bottom-up’ approach. I locate a critical juncture in the immigration debate during the early 1970s and argue that the shift towards the focus on illegality as a point of concern was due to an alignment of interests that brought an array of civil society organizations commonly understood as progressive to coincide with sectors of the bureaucracy.     

Reflexivity,tradition and racism in a former mining town

In this article I examine the formation of white working-class racialized discourses in a former coal-mining town in Leicestershire. I draw upon my ethnographic research in the area to explore the conditions in which young white adults confront, challenge and question what they perceive to be others’ conventional racist attitudes and beliefs, for example, the racist attitudes of members of their nuclear and extended families, friends and acquaintances. I argue that it is vital to examine these “moments of questioning” if white hegemony in predominantly working-class areas is to be understood.     

A white side of black Britain: The concept of racial literacy

Opposition to transracial adoption on both sides of the Atlantic, has been based, in part, on the assumption that white parents cannot understand race or racism and thus cannot properly prepare children of multiracial heritage to cope with racism. In this article I draw on a seven-year ethnographic study to offer an intensive case study of white transracial birth parents that counters this racial logic. I draw on a subset of data collected from field research and in-depth interviews with 102 members of black-white interracial families in England. I provide an analysis of three practices that I discovered among white transracial birth parents who were attempting to cultivate ‘black’ identities in their children of multiracial heritage. I offer the concept of ‘racial literacy’ to theorize their parental labour as a type of anti-racist project that remains under the radar of conventional sociological analyses of racism and anti-racist social movements.     

Ultrasonication of insulin-loaded microgel particles produced by internal gelation: Impact on particle's size and insulin bioactivity

Alginate-dextran sulfate (ADS) microgel has been used to protect insulin from gastrointestinal attack and as a carrier to promote insulin permeation through intestinal epithelium. The throughput of ADS submicron particles generation by emulsification/internal gelation is limited by its wide size distribution.     

Rangewide landscape genetics of an endemic Pacific northwestern salamander

A species' genetic structure often varies in response to ecological and landscape processes that differ throughout the species' geographic range, yet landscape genetics studies are rarely spatially replicated. The Cope's giant salamander (Dicamptodon copei) is a neotenic, dispersal‐limited amphibian with a restricted geographic range in the Pacific northwestern USA. We investigated which landscape factors affect D. copei gene flow in three regions spanning the species' range, which vary in climate, landcover and degree of anthropogenic disturbance. Least cost paths and Circuitscape resistance analyses revealed that gene flow patterns vary across the species' range, with unique combinations of landscape variables affecting gene flow in different regions. Populations in the northern coastal portions of the range had relatively high gene flow, largely facilitated by stream and river networks. Near the southeastern edge of the species' range, gene flow was more restricted overall, with relatively less facilitation by streams and more limitation by heat load index and fragmented forest cover. These results suggested that the landscape is more difficult for individuals to disperse through at the southeastern edge of the species' range, with terrestrial habitat desiccation factors becoming more limiting to gene flow. We suggest that caution be used when attempting to extrapolate landscape genetic models and conservation measures from one portion of a species' range to another.     

The altitudinal limit of Leptohyphes Eaton, 1882 and Lachlania Hagen, 1868 (Ephemeroptera: Leptohyphidae,Oligoneuriidae) in Ecuadorian Andes streams: searching for mechanisms

We explored mechanisms determining the upper altitudinal limit of ephemeropterans from two different genera: Leptohyphes Eaton, 1882 Eaton, A.E. (1882), ?An Announcement of New Genera of the Ephemeridae?, The Entomologist's Monthly Magazine, 18, 207–208. [Google Scholar] (Leptohyphidae) and Lachlania Hagen, 1868 Hagen, H.A. (1868), ?On Lachlania Abnormis: A New Genus and Species from Cuba Belonging to the Ephemerina?, Proceedings of the Boston Society of Natural History, 11, 372–375. [Google Scholar] (Oligoneuriidae). For this, we (1) surveyed the two taxa in 165 stream sites along a wide altitudinal gradient; (2) sampled benthic fauna at short altitudinal intervals along a stream, from 2780 to 3150 m above sea level; (3) collected adults at the lowest and highest sites; and (4) transplanted nymphs from the lowest to the highest study site in our stream to determine survival over time. Densities of the two taxa declined gradually with altitude and both disappeared between 2950 and 3080 m a.s.l. The upper altitudinal limit in the stream seemed to be most closely related to mean oxygen saturation, temperature, and current velocity. Adults were collected where the nymphs were found, but not at the upstream site where the nymphs were absent, implying limited upstream dispersal of adults and some of the altitudinal constraint lying at the adult stage. Short-term survival of transplanted nymphs was lower than that of controls, suggesting that the distribution was limited at the juvenile stage, and that at least some of the altitudinal constraint is related to the abiotic stream environment.