Showing papers by "Mark E. Cooper published in 2008"

Intensive blood glucose control and vascular outcomes in patients with type 2 diabetes.

Abstract: BACKGROUND: In patients with type 2 diabetes, the effects of intensive glucose control on vascular outcomes remain uncertain. METHODS: We randomly assigned 11,140 patients with type 2 diabetes to undergo either standard glucose control or intensive glucose control, defined as the use of gliclazide (modified release) plus other drugs as required to achieve a glycated hemoglobin value of 6.5% or less. Primary end points were composites of major macrovascular events (death from cardiovascular causes, nonfatal myocardial infarction, or nonfatal stroke) and major microvascular events (new or worsening nephropathy or retinopathy), assessed both jointly and separately. RESULTS: After a median of 5 years of follow-up, the mean glycated hemoglobin level was lower in the intensive-control group (6.5%) than in the standard-control group (7.3%). Intensive control reduced the incidence of combined major macrovascular and microvascular events (18.1%, vs. 20.0% with standard control; hazard ratio, 0.90; 95% confidence interval [CI], 0.82 to 0.98; P=0.01), as well as that of major microvascular events (9.4% vs. 10.9%; hazard ratio, 0.86; 95% CI, 0.77 to 0.97; P=0.01), primarily because of a reduction in the incidence of nephropathy (4.1% vs. 5.2%; hazard ratio, 0.79; 95% CI, 0.66 to 0.93; P=0.006), with no significant effect on retinopathy (P=0.50). There were no significant effects of the type of glucose control on major macrovascular events (hazard ratio with intensive control, 0.94; 95% CI, 0.84 to 1.06; P=0.32), death from cardiovascular causes (hazard ratio with intensive control, 0.88; 95% CI, 0.74 to 1.04; P=0.12), or death from any cause (hazard ratio with intensive control, 0.93; 95% CI, 0.83 to 1.06; P=0.28). Severe hypoglycemia, although uncommon, was more common in the intensive-control group (2.7%, vs. 1.5% in the standard-control group; hazard ratio, 1.86; 95% CI, 1.42 to 2.40; P<0.001). CONCLUSIONS: A strategy of intensive glucose control, involving gliclazide (modified release) and other drugs as required, that lowered the glycated hemoglobin value to 6.5% yielded a 10% relative reduction in the combined outcome of major macrovascular and microvascular events, primarily as a consequence of a 21% relative reduction in nephropathy. (ClinicalTrials.gov number, NCT00145925.)

Oxidative Stress as a Major Culprit in Kidney Disease in Diabetes

Abstract: It is postulated that localized tissue oxidative stress is a key component in the development of diabetic nephropathy. There remains controversy, however, as to whether this is an early link between hyperglycemia and renal disease or develops as a consequence of other primary pathogenic mechanisms. In the kidney, a number of pathways that generate reactive oxygen species (ROS) such as glycolysis, specific defects in the polyol pathway, uncoupling of nitric oxide synthase, xanthine oxidase, NAD(P)H oxidase, and advanced glycation have been identified as potentially major contributors to the pathogenesis of diabetic kidney disease. In addition, a unifying hypothesis has been proposed whereby mitochondrial production of ROS in response to chronic hyperglycemia may be the key initiator for each of these pathogenic pathways. This postulate emphasizes the importance of mitochondrial dysfunction in the progression and development of diabetes complications including nephropathy. A mystery remains, however, as to why antioxidants per se have demonstrated minimal renoprotection in humans despite positive preclinical research findings. It is likely that the utility of current study approaches, such as vitamin use, may not be the ideal antioxidant strategy in human diabetic nephropathy. There is now an increasing body of data to suggest that strategies involving a more targeted antioxidant approach, using agents that penetrate specific cellular compartments, may be the elusive additive therapy required to further optimize renoprotection in diabetes.

Transient high glucose causes persistent epigenetic changes and altered gene expression during subsequent normoglycemia

Abstract: The current goal of diabetes therapy is to reduce time-averaged mean levels of glycemia, measured as HbA1c, to prevent diabetic complications. However, HbA1c only explains <25% of the variation in risk of developing complications. Because HbA1c does not correlate with glycemic variability when adjusted for mean blood glucose, we hypothesized that transient spikes of hyperglycemia may be an HbA1c–independent risk factor for diabetic complications. We show that transient hyperglycemia induces long-lasting activating epigenetic changes in the promoter of the nuclear factor κB (NF-κB) subunit p65 in aortic endothelial cells both in vitro and in nondiabetic mice, which cause increased p65 gene expression. Both the epigenetic changes and the gene expression changes persist for at least 6 d of subsequent normal glycemia, as do NF-κB–induced increases in monocyte chemoattractant protein 1 and vascular cell adhesion molecule 1 expression. Hyperglycemia-induced epigenetic changes and increased p65 expression are prevented by reducing mitochondrial superoxide production or superoxide-induced α-oxoaldehydes. These results highlight the dramatic and long-lasting effects that short-term hyperglycemic spikes can have on vascular cells and suggest that transient spikes of hyperglycemia may be an HbA1c–independent risk factor for diabetic complications.

The role of advanced glycation end products in progression and complications of diabetes

Abstract: Context: Diabetic complications appear to be multifactorial in origin, but in particular, the biochemical process of advanced glycation, which is accelerated in diabetes as a result of chronic hyperglycemia and increased oxidative stress, has been postulated to play a central role in these disorders. Advanced glycation involves the generation of a heterogenous group of chemical moieties known as advanced glycated end products (AGEs), this reaction occurring as a result of a nonenzymatic reaction with glucose interacting with proteins, lipids, and nucleic acids, and involves key intermediates such as methylglyoxal. Evidence Synthesis: In this review we report on how these AGEs may exert deleterious effects in diabetes, as well as address current strategies to interrupt the formation or action of AGEs. First, AGEs act directly to induce cross-linking of long-lived proteins such as collagen to promote vascular stiffness, and, thus, alter vascular structure and function. Second, AGEs can interact with certain...

Oxidative Stress as a Major Culprit in Kidney Disease in

Abstract: It is postulated that localized tissue oxidative stress is a key component in the development of diabetic nephropathy. There remains controversy, however, as to whether this is an early link between hyperglycemia and renal disease or develops as a consequence of other primary pathogenic mechanisms. In the kidney, a number of pathways that generate reactive oxygen species (ROS) such as glycolysis, specific defects in the polyol pathway, uncoupling of nitric oxide synthase, xanthine oxidase, NAD(P)H oxidase, and advanced glycation have been identified as potentially major contributors to the pathogenesis of diabetic kidney disease. In addition, a unifying hypothesis has been proposed whereby mitochondrial production of ROS in response to chronic hyperglycemia may be the key initiator for each of these pathogenic pathways. This postulate emphasizes the importance of mitochondrial dysfunction in the progression and development of diabetes complications including nephropathy. A mystery remains, however, as to why antioxidants per se have demonstrated minimal renoprotection in humans despite positive preclinical research findings. It is likely that the utility of current study approaches, such as vitamin use, may not be the ideal antioxidant strategy in human diabetic nephropathy. There is now an increasing body of data to suggest that strategies involving a more targeted antioxidant approach, using agents that penetrate specific cellular compartments, may be the elusive additive therapy required to further optimize renoprotection in diabetes. Diabetes 57:1446‐1454, 2008

Receptor for Advanced Glycation End Products (RAGE) Deficiency Attenuates the Development of Atherosclerosis in Diabetes

Abstract: OBJECTIVE— Activation of the receptor for advanced glycation end products (RAGE) in diabetic vasculature is considered to be a key mediator of atherogenesis. This study examines the effects of deletion of RAGE on the development of atherosclerosis in the diabetic apoE−/− model of accelerated atherosclerosis. RESEARCH DESIGN AND METHODS— ApoE−/− and RAGE−/−/apoE−/− double knockout mice were rendered diabetic with streptozotocin and followed for 20 weeks, at which time plaque accumulation was assessed by en face analysis. RESULTS— Although diabetic apoE−/− mice showed increased plaque accumulation (14.9 ± 1.7%), diabetic RAGE−/−/apoE−/− mice had significantly reduced atherosclerotic plaque area (4.9 ± 0.4%) to levels not significantly different from control apoE−/− mice (4.3 ± 0.4%). These beneficial effects on the vasculature were associated with attenuation of leukocyte recruitment; decreased expression of proinflammatory mediators, including the nuclear factor-κB subunit p65 , VCAM-1 , and MCP-1 ; and reduced oxidative stress, as reflected by staining for nitrotyrosine and reduced expression of various NADPH oxidase subunits, gp91phox , p47phox , and rac-1 . Both RAGE and RAGE ligands, including S100A8/A9, high mobility group box 1 (HMGB1), and the advanced glycation end product (AGE) carboxymethyllysine were increased in plaques from diabetic apoE−/− mice. Furthermore, the accumulation of AGEs and other ligands to RAGE was reduced in diabetic RAGE−/−/apoE−/− mice. CONCLUSIONS— This study provides evidence for RAGE playing a central role in the development of accelerated atherosclerosis associated with diabetes. These findings emphasize the potential utility of strategies targeting RAGE activation in the prevention and treatment of diabetic macrovascular complications.

Inhibition of NADPH oxidase prevents advanced glycation end product-mediated damage in diabetic nephropathy through a protein kinase C-alpha-dependent pathway.

Abstract: OBJECTIVE —Excessive production of reactive oxygen species (ROS) via NADPH oxidase has been implicated in the pathogenesis of diabetic nephropathy. Since NADPH oxidase activation is closely linked to other putative pathways, its interaction with changes in protein kinase C (PKC) and increased advanced glycation was examined. RESEARCH DESIGN AND METHODS —Streptozotocin-induced diabetic or nondiabetic Sprague Dawley rats were followed for 32 weeks, with groups randomized to no treatment or the NADPH oxidase assembly inhibitor apocynin (15 mg · kg −1 · day −1 ; weeks 16–32). Complementary in vitro studies were performed in which primary rat mesangial cells, in the presence and absence of advanced glycation end products (AGEs)-BSA, were treated with either apocynin or the PKC-α inhibitor Ro-32-0432. RESULTS —Apocynin attenuated diabetes-associated increases in albuminuria and glomerulosclerosis. Circulating, renal cytosolic, and skin collagen–associated AGE levels in diabetic rats were not reduced by apocynin. Diabetes-induced translocation of PKC, specifically PKC-α to renal membranes, was associated with increased NADPH-dependent superoxide production and elevated renal, serum, and urinary vascular endothelial growth factor (VEGF) concentrations. In both diabetic rodents and in AGE-treated mesangial cells, blockade of NADPH oxidase or PKC-α attenuated cytosolic superoxide and PKC activation and increased VEGF. Finally, renal extracellular matrix accumulation of fibronectin and collagen IV was decreased by apocynin. CONCLUSIONS —In the context of these and previous findings by our group, we conclude that activation of NADPH oxidase via phosphorylation of PKC-α is downstream of the AGE–receptor for AGE interaction in diabetic renal disease and may provide a novel therapeutic target for diabetic nephropathy.

Nanomechanical detection of antibiotic-mucopeptide binding in a model for superbug drug resistance.

Abstract: The alarming growth of the antibiotic-resistant superbugs methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE) is driving the development of new technologies to investigate antibiotics and their modes of action. We report the label-free detection of vancomycin binding to bacterial cell wall precursor analogues (mucopeptides) on cantilever arrays, with 10 nM sensitivity and at clinically relevant concentrations in blood serum. Differential measurements have quantified binding constants for vancomycin-sensitive and vancomycin-resistant mucopeptide analogues. Moreover, by systematically modifying the mucopeptide density we gain new insights into the origin of surface stress. We propose that stress is a product of a local chemical binding factor and a geometrical factor describing the mechanical connectivity of regions activated by local binding in terms of a percolation process. Our findings place BioMEMS devices in a new class of percolative systems. The percolation concept will underpin the design of devices and coatings to significantly lower the drug detection limit and may also have an impact on our understanding of antibiotic drug action in bacteria.

UKPDS and the Legacy Effect

Abstract: The United Kingdom Prospective Diabetes Study (UKPDS) continues to produce important evidence concerning the evolution of type 2 diabetes and its management. Two studies published in this issue of the Journal provide some answers to two questions of fundamental importance to patients with diabetes and to physicians alike. In one article, Holman et al. (UKPDS 80)1 provide data that confirm a so-called legacy effect associated with intensive glucose control in patients with type 2 diabetes, long after the cessation of randomized intervention. This finding provides a fitting parallel to the observations of the Diabetes Control and Complications Trial/Epidemiology of Diabetes . . .

Interlaboratory comparison study of calibration standards for foraminiferal Mg/Ca thermometry

Abstract: An interlaboratory study of Mg/Ca and Sr/Ca ratios in three commercially available carbonate reference materials (BAM RS3, CMSI 1767, and ECRM 752-1) was performed with the participation of 25 laboratories that determine foraminiferal Mg/Ca ratios worldwide. These reference materials containing Mg/Ca in the range of foraminiferal calcite (0.8 mmol/mol to 6 mmol/mol) were circulated with a dissolution protocol for analysis. Participants were asked to make replicate dissolutions of the powdered samples and to analyze them using the instruments and calibration standards routinely used in their laboratories. Statistical analysis was performed in accordance with the International Standardization Organization standard 5725, which is based on the analysis of variance (ANOVA) technique. Repeatability (RSDr%), an indicator of intralaboratory precision, for Mg/Ca determinations in solutions after centrifuging increased with decreasing Mg/Ca, ranging from 0.78% at Mg/Ca = 5.56 mmol/mol to 1.15% at Mg/Ca = 0.79 mmol/mol. Reproducibility (RSDR%), an indicator of the interlaboratory method precision, for Mg/Ca determinations in centrifuged solutions was noticeably worse than repeatability, ranging from 4.5% at Mg/Ca = 5.56 mmol/mol to 8.7% at Mg/Ca = 0.79 mmol/mol. Results of this study show that interlaboratory variability is dominated by inconsistencies among instrument calibrations and highlight the need to improve interlaboratory compatibility. Additionally, the study confirmed the suitability of these solid standards as reference materials for foraminiferal Mg/Ca (and Sr/Ca) determinations, provided that appropriate procedures are adopted to minimize and to monitor possible contamination from silicate mineral phases.

ACE2 Deficiency Modifies Renoprotection Afforded by ACE Inhibition in Experimental Diabetes

Abstract: Objective: The degradation of angiotensin (Ang) II by angiotensin converting enzyme 2 (ACE2), leading to the formation of Ang 1–7, is an important step in the renin-angiotensin system (RAS), and one which is significantly altered in the diabetic kidney. This study examines the role of ACE2 in early renal changes associated with diabetes, and the influence of ACE2 deficiency on ACE inhibitor-mediated renoprotection. Methods: Streptozotocin diabetes was induced in male c57bl6 mice and ACE2 KO mice. After five weeks of study, animals were randomised to receive the ACE inhibitor, perindopril (2 mg/kg/day). Wild-type mice were further randomised to receive the selective ACE2 inhibitor, MLN-4760 (10 mg/kg/day) and followed for an additional five weeks. Markers of renal function and injury were then assessed. Results: The induction of diabetes in wild-type mice was associated with a reduction in renal ACE2 expression and decreased Ang1-7. In diabetic mice receiving MLN-4760 and in ACE2 KO mice, diabetes-associated albuminuria was enhanced, associated with an increase in blood pressure. However, renal hypertrophy and fibrogenesis were reduced in diabetic mice with ACE2 deficiency and hyperfiltration was attenuated. Diabetic wild type mice treated with an ACE inhibitor experienced a reduction in albuminuria and blood pressure. These responses were attenuated in both diabetic ACE2 KO mice and diabetic mice receiving MLN-4760. However, other renoprotective and antifibrotic actions of ACE inhibition in diabetes were preserved in ACE2 deficient mice. Conclusions: The expression of ACE2 is significantly modified by diabetes, which impacts both on the pathogenesis of kidney disease, as well as the responsiveness to RAS blockade. These data indicate that ACE2 is a complex, and site-specific modulator of diabetic kidney disease.

The role of AGEs in cardiovascular disease.

Abstract: Advanced glycation end-products (AGEs) are generated in the diabetic milieu, as a result of chronic hyperglycemia and enhanced oxidative stress. These AGEs, via direct and receptor dependent pathways promote the development and progression of cardiovascular disease. AGEs accumulate at many sites of the body including the heart and large blood vessels in diabetes. These modified proteins interact with receptors such as RAGE to induce oxidative stress, increase inflammation by promoting NFkappaB activation and enhance extracellular matrix accumulation. These biological effects translate to accelerated plaque formation in diabetes as well as increased cardiac fibrosis with consequent effects on cardiac function. Strategies to reduce the ligation of AGEs to their receptors such as agents which reduce AGE accumulation, soluble RAGE which acts as a competitive antagonist to the binding of AGEs to RAGE and genetic deletions of RAGE appear to attenuate diabetes associated atherosclerosis. Benefits on cardiac dysfunction with these inhibitors of the AGE/RAGE axis are not as well characterised. In conclusion, therapeutic strategies targeting AGEs appear to have significant clinical potential, often in combination with currently used agents such as inhibitors of the renin-angiotensin system, to reduce the major burden of diabetes, its associated cardiovascular complications.

Cardiac inflammation associated with a Western diet is mediated via activation of RAGE by AGEs

Abstract: A diet high in fat induces cardiac hypertrophy, inflammation, and oxidative stress. Although such actions have largely been ascribed to fat deposition, the accumulation of advanced glycation end pr...

ACE gene polymorphism and losartan treatment in type 2 diabetic patients with nephropathy.

Abstract: Losartan treatment reduced renal outcomes in proteinuric patients with type 2 diabetes in the Reduction of Endpoints in NIDDM with the Angiotensin II Antagonist Losartan (RENAAL) study. It is unknown whether an insertion (I)/deletion (D) polymorphism in the angiotensin I-converting enzyme (ACE) gene predicts renal outcomes and death and influences the effect of losartan in these patients. Pharmacogenetic analyses were performed comparing losartan with placebo administered with conventional blood pressure-lowering therapy in 1435 (95%) of the 1513 RENAAL study patients. The primary endpoint was the composite of doubling of baseline serum creatinine concentration, end-stage renal disease (ESRD) or death. Cox regression models were stratified on baseline proteinuria and included treatment, geographic region, ACE/ID genotype, and treatment x genotype interaction. Within the placebo group, subjects with the ID or DD genotype were more likely than those with the II genotype to reach the composite endpoint (by 17.5% and 38.1%, respectively, P = 0.029) or its individual components. Within the losartan group, genotype did not correlate with reaching the composite endpoint. Compared with placebo, however, losartan reduced the risk of reaching the composite endpoint by 5.8% (95% confidence interval, -23.3, 28.0), 17.6% (3.8, 29.4), and 27.9% (7.0, 44.1) among those with the II, ID, and DD genotypes, respectively. Similar trends were demonstrated for the individual endpoints. In conclusion, proteinuric type 2 diabetic patients with the D allele of the ACE gene have an unfavorable renal prognosis, which can be mitigated and even improved by losartan.

Heat shock protein expression in diabetic nephropathy.

Abstract: Heat shock protein (HSP) HSP27, HSP60, HSP70, and HSP90 are induced by cellular stresses and play a key role in cytoprotection. Both hyperglycemia and glomerular hypertension are crucial determinants in the pathogenesis of diabetic nephropathy and impose cellular stresses on renal target cells. We studied both the expression and the phosphorylation state of HSP27, HSP60, HSP70, and HSP90 in vivo in rats made diabetic with streptozotocin and in vitro in mesangial cells and podocytes exposed to either high glucose or mechanical stretch. Diabetic and control animals were studied 4, 12, and 24 wk after the onset of diabetes. Immunohistochemical analysis revealed an overexpression of HSP25, HSP60, and HSP72 in the diabetic outer medulla, whereas no differences were seen in the glomeruli. Similarly, exposure neither to high glucose nor to stretch altered HSP expression in mesangial cells and podocytes. By contrast, the phosphorylated form of HSP27 was enhanced in the glomerular podocytes of diabetic animals, and in vitro exposure of podocytes to stretch induced HSP27 phosphorylation via a P38-dependent mechanism. In conclusion, diabetes and diabetes-related insults differentially modulate HSP27, HSP60, and HSP70 expression/phosphorylation in the glomeruli and in the medulla, and this may affect the ability of renal cells to mount an effective cytoprotective response.

A high sensitivity assay for the inflammatory marker C-Reactive protein employing acoustic biosensing.

Abstract: C-Reactive Protein (CRP) is an acute phase reactant routinely used as a biomarker to assess either infection or inflammatory processes such as autoimmune diseases. CRP also has demonstrated utility as a predictive marker of future risk of cardiovascular disease. A new method of immunoassay for the detection of C-Reactive Protein has been developed using Resonant Acoustic Profiling™ (RAP™) with comparable sensitivity to a high sensitivity CRP ELISA (hsCRP) but with considerable time efficiency (12 minutes turnaround time to result). In one method, standard solutions of CRP (0 to 231 ng/mL) or diluted spiked horse serum sample are injected through two sensor channels of a RAP™ biosensor. One contains a surface with sheep antibody to CRP, the other a control surface containing purified Sheep IgG. At the end of a 5-minute injection the initial rate of change in resonant frequency was proportional to CRP concentration. The initial rates of a second sandwich step of anti-CRP binding were also proportional to the sample CRP concentration and provided a more sensitive method for quantification of CRP. The lower limit of detection for the direct assay and the homogenous sandwich assay were both 20 ng/mL whereas for the direct sandwich assay the lower limit was 3 ng/mL. In a step towards a rapid clinical assay, diluted horse blood spiked with human CRP was passed over one sensor channel whilst a reference standard solution at the borderline cardiovascular risk level was passed over the other. A semi-quantities ratio was thus obtained indicative of sample CRP status. Overall, the present study revealed that CRP concentrations in serum that might be expected in both normal and pathological conditions can be detected in a time-efficient, label-free immunoassay with RAP™ detection technology with determined CRP concentrations in close agreement with those determined using a commercially available high sensitivity ELISA.

Angiotensin converting enzyme 2 in the kidney

Abstract: 1. Angiotensin converting enzyme 2 (ACE2) is an important homeostatic component of the renin angiotensin system (RAS). ACE2 both degrades the vasoconstrictor, angiotensin II and generates the potent vasodilator peptide, angiotensin 1-7. These actions counterbalance those of ACE. 2. ACE2 is highly expressed in the healthy kidney, particularly in the proximal tubules, where it colocalizes with ACE and angiotensin receptors. 3. Kidney disease and subtotal nephrectomy is associated with a reduction in renal ACE2 expression, possibly facilitating the damaging effects of angiotensin II in the failing kidney. Acquired or genetic ACE2 deficiency also appears to exacerbate renal damage and albuminuria in experimental models, supporting this hypothesis. 4. ACE2 also has an important role in blood pressure control. Many models of hypertension are associated with reduced ACE2 expression. Although ACE2 KO animals are normotensive, in states associated with activation of the RAS, ACE2 overexpression improves blood pressure control and reduces angiotensin responsiveness.

Genotypic variation for drought stress response traits in soybean. I. Variation in soybean and wild Glycine spp. for epidermal conductance, osmotic potential, and relative water content

Abstract: Studies were undertaken to assess genotypic variation in soybean and related wild species for traits with putative effects on leaf turgor maintenance in droughted plants. Traits of interest were (i) epidermal conductance (ge) which influences the rate of water loss from stressed leaves after stomatal closure; (ii) osmotic adjustment (OA) as indicated by tissue osmotic potential (π), which potentially affects the capacity to withdraw water at low soil water potential; and (iii) relative water content (RWC) at incipient leaf death (critical relative water content, RWCC), which is a measure of the dehydration tolerance of leaf tissue. The germplasm comprised a diverse set of 58 soybean genotypes, 2 genotypes of the annual wild species G. soja and 9 genotypes representing 6 perennial wild Glycine spp. indigenous/endemic to Australia. Seedling plants were grown in soil-filled beds in the glasshouse and exposed to terminal water deficit stress from the second trifoliolate leaflet stage (21 days after sowing). Measurements were made on well watered plants, moderately stressed plants, and at incipient plant death, in 2 separate studies. In both studies, there were significant genotypic differences in all 3 traits in the stressed plants. However, across the 3 sample times, ge decreased and the absolute magnitude of π increased, indicating that the expression of these traits changed as the plants acclimated to the stress. RWC was therefore used as a covariate to adjust the genotypic values of π and ge in order to facilitate comparison at a consistent plant water status of 70% RWC. There was statistically significant genotypic variation for the adjusted values, ge70 and π70, in both studies, and genotypic correlations between the 2 studies were significant (P < 0.05) and positive for all 3 traits: ge70 (r = 0.48), π70 (r = 0.50), and RWCC (r = 0.53). Among the soybean genotypes, there was at least a 2-fold range in ge70, a 0.7 MPa range in π70, and a 12 percentage point range in RWCC. Some of the perennial wild genotypes exhibited lower values of ge and RWCC and greater OA than soybean and G. soja, consistent with adaptation to drier environments. While the repeatability of measurement between experiments was variable among genotypes, the studies confirmed the existence of genotypic differences for ge, OA, and RWCC in cultivated soybean, with a wider range among the wild germplasm.

Genotypic variation for drought stress response traits in soybean. II. Inter-relations between epidermal conductance, osmotic potential, relative water content, and plant survival

Abstract: As part of a project exploring the potential for using leaf physiological traits to improve drought tolerance in soybean, studies were conducted to explore whether epidermal conductance (ge), osmotic potential (π), and relative water content (RWC) influenced turgor maintenance and ultimately the survival of droughted plants. In a glasshouse study, plants of 8 soybean genotypes that showed different expression of the traits were grown in well watered soil-filled beds for 21 days and then exposed to terminal water deficit stress. The trends in each trait were then monitored periodically until plant death. Genotypic differences were observed in the rate of decline in RWC as the soil dried, in the temporal patterns of change in ge and π, in the duration of survival after watering ceased, and in the critical relative water content (RWCC) at which plants died. In general, ge became smaller and π became more negative as RWC declined and plants acclimated to the increasing stress. Genotypic differences in ge remained broadly consistent as RWC declined. In contrast, the genotypic rankings for π in stressed plants were poorly correlated with those for well watered plants, indicating differential genotypic capacity for osmotic adjustment (OA) in response to stress. Survival times among genotypes after stress commenced ranged from 27 to 41 days, while RWCC ranged from 49% down to 41%. The differences in survival time among the genotypes were able to be explained by genotypic differences in the rate of decline in RWC and in the RWCC, using a multiple linear regression relationship (R 2 = 0.94**). In turn, genotypic differences in the rate of decline in RWC were positively correlated (r = 0.75*) with ge at 70% RWC, and with OA over the drying period (r = 0.98**). In a second study in a controlled environment facility, leaf area retention at 90% soil water extraction was greatest in the one genotype that combined low ge, high OA, and low RWCC. Overall, the responses from the two studies were consistent with the hypothesis that turgor maintenance and ultimately leaf and plant survival of different genotypes during advanced stages of drought stress are enhanced by low ge, high OA capacity, and low RWCC.

Direct acoustic profiling of DNA hybridisation using HSV type 1 viral sequences

Abstract: We describe the detection of specific, conserved DNA sequences of herpes simplex virus (HSV) type 1 by means of a novel, high sensitivity acoustic biosensor. Repeated assays on planar and polymeric carboxylic acid- and biotin-presenting surface chemistries enabled statistical comparison of assay specificity and sensitivity and evaluation of assayZ-factor scores. Using a three minute hybridisation with NeutrAvidin capture for signal enhancement, it was possible to detect HSV viral nucleic acids at 5.2 × 10−11 M concentration.

Agents in development for the treatment of diabetic nephropathy.

Abstract: Background: Nephropathy is a major cause of morbidity and mortality in diabetic patients. Current treatments include optimization of glycemic and blood pressure control, but more innovative strategies are needed for the prevention and treatment of diabetic nephropathy. Objectives: To review emerging therapies for diabetic nephropathy. Methods: This paper discusses the molecular mechanisms of diabetic nephropathy and the potential therapeutic interventions. Results/conclusion: New therapies, including those targeting the accumulation of advanced glycation end products (AGEs) and reactive oxygen species (ROS) generation, are likely to feature in future treatment regimens. Other approaches that at this stage do not appear to be progressing include the glycosaminoglycan sulodexide and the protein kinase C-beta (PKC-β) inhibitor, ruboxistaurin.

Therapeutic interruption of advanced glycation in diabetic nephropathy: do all roads lead to Rome?

Abstract: A major common feature of the chemically disparate compounds that inhibit advanced glycation end product (AGE) accumulation or signaling is their ability to show end-organ protection in experimental models of diabetes complications. The mechanisms by which these AGE- lowering therapies confer their benefits remain unsolved. Is it the reduction in tissue AGE levels per se or the inhibition of downstream signal transduction (as has been described with the soluble receptor for AGE)? Possible modes of action that need to be investigated include the ability of some of these agents to stimulate antioxidant defenses, to lower cholesterol and other lipid levels, and to inhibit low-grade inflammation. To understand these novel mechanisms, further examination of the advanced glycation pathway and, in particular, the diverse action of these agents in ameliorating the development of diabetic complications is needed.

Diabetic patients and kidney protection: an attainable target.

Abstract: Diabetic nephropathy, the major cause of end-stage renal disease in the world occurs as a result of both metabolic and haemodynamic insults, thus emphasizing the importance of optimizing glycaemic and blood pressure control in patients with or at risk of this disorder. The mainstay of antihypertensive therapy is now inhibition of the renin-angiotensisn system involving the use of angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers. The appropriate blood pressure level for the commencement of these drugs and what should be the achieved blood pressure in individuals with diabetes remain controversial. Promising new therapies are currently under preclinical investigation or in early stage clinical trials, and hopefully these newer agents, probably used as adjunct therapies, will further improve the prognosis of individuals with diabetes with early or overt renal disease.

Effect of LDL Cholesterol and Treatment With Losartan on End-Stage Renal Disease in the RENAAL Study

Abstract: Renal pathology and dyslipidemia commonly coexist. Treatments that lower albuminuria/proteinuria may lower lipids, but it is not known whether lipid lowering independent of lessening albuminuria/proteinuria slows progression of kidney disease. We examined the association between LDL cholesterol levels and treatment with losartan on end-stage renal disease (ESRD). Lipid levels and albuminuria measurements were obtained at baseline and at year 1 in a post hoc analysis from the Reduction of Endpoints in NIDDM with the Angiotensin II Antagonist Losartan (RENAAL) study, which compared the effects of losartan- versus placebo-based antihypertensive therapy in patients with type 2 diabetes and nephropathy. LDL cholesterol lowering was associated with a lower risk of ESRD; however, this seemed to be largely an association with the reduction in albuminuria.

Genotypic variation for drought stress response traits in soybean. III. Broad-sense heritability of epidermal conductance, osmotic potential, and relative water content

Abstract: The broad-sense heritability of 3 traits related to leaf survival in severely stressed plants was studied in several hybrid soybean populations. The 3 traits were epidermal conductance (ge), osmotic potential (π), and relative water content (RWC). The populations were generated by hybridising unrelated parental genotypes previously shown to differ in the 3 traits. ge (mm/s) was measured on well watered plants from 10 populations involving all combinations of 5 parental lines, grown in soil-filled beds in the glasshouse. π (MPa) and RWC (%) were measured on severely stressed plants of 3 populations involving all combinations of 3 different parents, growing into a terminal water deficit under a rainout shelter in the field. Broad-sense heritability for ge was significantly different from zero (P < 0.05) in all 10 populations and ranged from 60% to 93%. Heritability estimates for π70 (the tissue osmotic potential at 70% RWC) ranged from 33% to 71%. Only two estimates were statistically significant (P < 0.05) because of large standard errors and the fact that parental differences were smaller than previously observed. Broad-sense heritability for RWC of severely stressed plants ranged from 40% to 74%, and was statistically significant (P < 0.05) for 2 of the 3 populations. For all 3 traits, F2 progeny distributions were consistent with quantitative inheritance with a high degree of additive gene action. It was concluded that capacity exists to breed varieties with low ge, low π70, and high RWC in stressed plants. However, in the case of osmotic potential, genotypes with lower π70 combined with greater precision of measurement would be needed than proved possible in these studies. Further, specific strategies would be needed to select for the critical RWC, the minimal RWC at which leaf tissues die and which provides a measure of tissue dehydration tolerance. More research is also needed to characterise the dynamic relations between ge, π, and RWC in influencing leaf survival in soybean, before they could be confidently used in a breeding program to improve drought tolerance.