H-ras, K-ras, and inner plasma membrane raft proteins operate in nanoclusters with differential dependence on the actin cytoskeleton

Plasma membrane compartmentalization imposes lateral segregation on membrane proteins that is important for regulating signal transduction. We use computational modeling of immunogold spatial point patterns on intact plasma membrane sheets to test different models of inner plasma membrane organization. We find compartmentalization at the nanoscale level but show that a classical raft model of preexisting stable domains into which lipid raft proteins partition is incompatible with the spatial point patterns generated by the immunogold labeling of a palmitoylated raft marker protein. Rather, approximate to 30% of the raft protein exists in cholesterol-dependent nanoclusters, with approximate to 70% distributed as monomers. The cluster/monomer ratio (number of proteins in clusters/number of proteins outside clusters) is independent of expression level. H-rasG12V and K-rasG12V proteins also operate in nanoclusters with fixed cluster/monomer ratios that are independent of expression level. Detailed calibration of the immunogold imaging protocol suggests that radii of raft and RasG12V protein nanoclusters may be as small as 11 and 6 nm, respectively, and shows that the nanoclusters contain small numbers (6.0-7.7) of proteins. Raft nanoclusters do not form if the actin cytoskeleton is disassembled. The formation of K-rasG12V but not H-rasG12V nanoclusters also is actin-dependent. K-rasG12V but not H-rasG12V signaling is abrogated by actin cytoskeleton disassembly, which shows that nanoclustering is critical for Ras function. These findings argue against stable preexisting domains on the inner plasma membrane in favor of dynamic actively regulated nanoclusters similar to those proposed for the outer plasma membrane. RasG12V nanoclusters may facilitate the assembly of essential signal transduction complexes.

The aldosterone-renin ratio in screening for primary aldosteronism

Recognition that primary aldosteronism (PAL) is a common specifically treatable form of hypertension and that most patients are normokalemic has led to a marked increase in demand for aldosterone/renin ratio (ARR) testing as a means of screening for this disorder. The value of this screening test depends on an appreciation of many factors (such as diet, posture, time of day, presence of hypokalemia, medications, age, and renal function), which can affect the results, on the care with which these factors are either controlled or their effects taken into account, and on access to reliable and reproducible assays for renin and aldosterone. Even then, physiological day-to-day variability reduces the value of a single estimation, and repeated testing is necessary before a decision that PAL is highly likely (warranting further testing) or highly unlikely can be made. Provided that testing of aldosterone suppressibility is always carried out to confirm or exclude the diagnosis, and the subtype is determined by hybrid gene testing and adrenal venous sampling, wide application of the ARR can have a major beneficial clinical impact with improved therapeutic outcomes, including possible cure in those with unilateral disease.

Impact of premorbid undernutrition on outcome in stroke patients

Background and Purpose - To assess the prevalence of premorbid undernutrition and its impact on outcomes 1 month after stroke. Methods - The study recruited from consecutive stroke admissions during a 10-month period. Premorbid nutritional status ( using the subjective global assessment [SGA]), premorbid functioning ( modified Rankin scale [MRS]), and stroke severity ( National Institutes of Health Stroke Scale [NIHSS] score) were assessed at admission. The associations between premorbid nutritional status, poor outcome ( defined as MRS greater than or equal to 3), and mortality were examined before and after adjustment for confounding variables, including age, gender, stroke risk factors, stroke severity, and admission serum albumin. Results - Thirty of 185 patients were assessed as having undernutrition at admission. Significant unadjusted associations were observed between undernutrition and poor outcome (odds ratio [OR], 3.4; 95% CI, 1.3 to 8.7; P = 0.01), and mortality (OR, 3.1, 95% CI, 1.3 to 7.7; P = 0.02) at 1 month. NIHSS, age, and premorbid MRS were also significantly associated with poor outcomes. After adjustment for these factors, the effect size of associations remained important but not significant ( poor outcome: OR, 2.4; 95% CI, 0.7 to 9.0, P = 0.18; mortality: OR, 3.2; 95% CI, 1.0 to 10.4, P = 0.05). Conclusions - Premorbid undernutrition, as assessed using the SGA, appears to be an independent predictor of poor stroke outcome. Stroke prevention strategies should target undernutrition in the population at risk for stroke to improve outcomes.

Three putative types of El Nino revealed by spatial variability in impact on Australian wheat yield

The El Nino-Southern Oscillation (ENSO) phenomenon significantly impacts rainfall and ensuing crop yields in many parts of the world. In Australia, El Nino events are often associated with severe drought conditions. However, El Nino events differ spatially and temporally in their manifestations and impacts, reducing the relevance of ENSO-based seasonal forecasts. In this analysis, three putative types of El Nino are identified among the 24 occurrences since the beginning of the twentieth century. The three types are based on coherent spatial patterns (footprints) found in the El Nino impact on Australian wheat yield. This bioindicator reveals aligned spatial patterns in rainfall anomalies, indicating linkage to atmospheric drivers. Analysis of the associated ocean-atmosphere dynamics identifies three types of El Nino differing in the timing of onset and location of major ocean temperature and atmospheric pressure anomalies. Potential causal mechanisms associated with these differences in anomaly patterns need to be investigated further using the increasing capabilities of general circulation models. Any improved predictability would be extremely valuable in forecasting effects of individual El Nino events on agricultural systems.

The ABC transporter gene in the sirodesmin biosynthetic gene cluster of Leptosphaeria maculans is not essential for sirodesmin production but facilitates self-protection

Epipolythiodioxopiperazine toxins are secreted by a range of fungi, including Leptosphaeria maculans, which produces sirodesmin, and Aspergillus fumigatus, which produces gliotoxin. The L. maculans biosynthetic gene cluster for sirodesmin includes an ABC transporter gene, sirA. Disruption of this gene led to increased secretion of sirodesmin into the medium and an altered ratio of sirodesmin to its immediate precursor. The transcription pattern of a peptide synthetase that catalyses an early step in sirodesmin biosynthesis was elevated in the sirA mutant by 47% over a 7-day period. This was consistent with the finding that the transporter mutant had elevated sirodesmin levels. Despite increased production of sirodesmin, the sit-A mutant was more sensitive to both sirodesmin and gliotoxin. The putative gliotoxin transporter gene, gliA, (a major facilitator superfamily transporter) from A.fumigatus complemented the tolerance of the L. maculans sirA mutant to gliotoxin, but not to sirodesmin. The results indicate that SirA contributes to self-protection against sirodesmin in L. maculans and suggest a transporter other than SirA is primarily responsible for efflux of endogenously produced sirodesmin. (C) 2004 Elsevier Inc. All rights reserved.

Clay mineralogical, geochemical and isotopic tracing of the evolution of the Woodleigh impact structure, Southern Carnarvon Basin, Western Australia

Chaotically structured diamictite from the inner ring syncline surrounding the central uplift of the Woodleigh impact structure contains shocked metamorphic and impact melt-rock fragments, largely derived from Ordovician and Devonian target sandstones. Coarse illite fractions (< 2 mu m) from the sandstones containing no K-feldspar yield K-Ar ages of around 400 Ma, whereas the K-Ar ages of authigenic clays of > 0.2 mu m fractions from the diamictite without smectite and K-feldspar cluster around 360 Ma, consistent with Rb-Sr data. Crystallisation of newly formed illite in the impact melt rock clasts and recrystallisation of earlier formed illite in the sandstone clasts preserved in the diamictite, are attributed to impact-induced hydrothermal processes in the Late Devonian. The illitic clays from the diamictite and from the sandstones have very similar trace element compositions, with significantly enriched incompatible lithophile elements, which increase in concentrations correlatively with those of the compatible ferromagnesian elements. The unusual trace element associations in the clays may be due to the involvement of hot gravity-driven basinal fluids that interacted with rocks of the Precambrian craton to the east of the study area, or with such material transported and reworked in the studied sedimentary succession.

The prevalence and impact of overweight and obesity in an Australian obstetric population

Objective: To assess the prevalence and impact of overweight and obesity in an Australian obstetric population. Design, setting and participants: The Mater Mother's Hospital (MMH), South Brisbane, is an urban tertiary referral maternity hospital. We reviewed data for the 18401 women who were booked for antenatal care at the MMH, delivered between January 1998 and December 2002, and had a singleton pregnancy. Of those women, 14 230 had an estimated pre-pregnancy body mass index (BMI) noted in their record; 2978 women with BMI 40 kg/m(2)). Main outcome measures: Prevalence of overweight and obesity in an obstetric population; maternal, peripartum and neonatal outcomes associated with raised BMI. Results: Of the 14230 women, 6443 (45%) were of normal weight, and 4809 (34%) were overweight, obese or morbidly obese. Overweight, obese and morbidly obese women were at increased risk of adverse outcomes (figures represent adjusted odds ratio [AOR] [95% Cl]): hypertensive disorders of pregnancy (overweight 1.74 [1.45-2.15], obese 3.00 [2.40-3.74], morbidly obese 4.87 [3.27-7.24]); gestational diabetes (overweight 1.78 [1.25-2.52], obese 2.95 [2.05-4.25], morbidly obese 7.44 [4.42-12.54]); hospital admission longer than 5 days (overweight 1.36 [1.13-1.63], obese 1.49 [1.21-1.86], morbidly obese 3.18 [2.19-4.61]); and caesarean section (overweight 1.50 [1.36-1.66], obese 2.02 [1.79-2.29], morbidly obese 2.54 [1.94-3.321). Neonates born to obese and morbidly obese women had an increased risk of birth defects (obese 1.58 (1.02-2.46], morbidly obese 3.41 [1.67-6.94]); and hypoglycaemia (obese 2.57 [1.39-4.78], morbidly obese 7.14 [3.04-16.74]). Neonates born to morbidly obese women were at increased risk of admission to intensive care (2.77 [1.81-4.25]); premature delivery (< 34 weeks' gestation) (2.13 [1.13-4.01]); and jaundice (1.44 [1.09-1.89]). Conclusions: Overweight and obesity are common in pregnant women. Increasing BMI is associated with maternal and neonatal outcomes that may increase the costs of obstetric care. To assist in planning health service delivery, we believe that BMI should be routinely recorded on perinatal data collection sheets