bluuebunny/crossref_metadata_2025 huggingface dataset:
Evidence for a Role of Transforming Growth Factor (TGF)-β1 in the Induction of Postglomerular Albuminuria in Diabetic Nephropathy
Authors: Russo Leileata M. , del Re Elisabetta , Brown Dennis , Lin Herbert Y.
Published: January 2007

Transforming growth factor-β (TGF-β) has previously been implicated in the progression of diabetic nephropathy, including the onset of fibrosis and albuminuria. Here we report for the first time the use of a high-affinity TGF-β1 binding molecule, the soluble human TGF-β type II receptor (sTβRII.Fc), in the treatment of diabetic nephropathy in 12-week streptozotocin-induced diabetic Sprague-Dawley rats. In vitro studies using immortalized rat proximal tubule cells revealed that 50 pmol/l TGF-β1 disrupted albumin uptake (P < 0.001 vs. control), an inhibition significantly reversed by the use of the sTβRII.Fc (1,200 pmol/l). In vivo studies demonstrated that treatment with sTβRII.Fc reduced urinary albumin excretion by 36% at 4 weeks, 59% at 8 weeks (P < 0.001), and 45% at 12 weeks (P < 0.01 for diabetic vs. treated). This was correlated with an increase in megalin expression (P < 0.05 for diabetic vs. treated) and a reduction in collagen IV expression following sTβRII.Fc treatment (P < 0.001 for diabetic vs. treated). These changes occurred independently of changes in blood glucose levels. This study demonstrates that the sTβRII.Fc is a potential new agent for the treatment of fibrosis and albuminuria in diabetic nephropathy and may reduce albuminuria by reducing TGF-β1–induced disruptions of renal proximal tubule cell uptake of albumin.
Genetic Reconstitution of Autoimmune Type 1 Diabetes With Two Major Susceptibility Genes in the Rat
Authors: Yokoi Norihide , Hayashi Chihiro , Fujiwara Yuuka , Wang He-Yao , Seino Susumu
Published: January 2007

The Komeda diabetes-prone (KDP) rat is an animal model of human autoimmune type 1 diabetes. We have previously shown that two major susceptibility genes, the major histocompatibility complex (MHC) RT1u haplotype and Cblb (Casitas B-lineage lymphoma b) mutation, are responsible for the development of diabetes in KDP rats, suggesting a two-gene model for development of the disease. To confirm the two-gene model, we produced a congenic strain carrying mutated Cblb alleles of the KDP rat on a non-KDP genetic background harboring the RT1u haplotype on its MHC. Despite the low incidence and delayed onset of diabetes, the congenic strain did develop the disease, indicating that type 1 diabetes can be reconstituted on a non-KDP genetic background with the RT1u haplotype and Cblb mutation. Similar to observations in KDP rats, the congenic strain showed insulitis and thyroiditis, symptoms of autoimmunity. The low incidence and delayed onset of the disease strongly suggest involvement of genetic modifiers; the congenic strain established in this study should be useful for the mapping and identification of such modifiers.
Women With Mitochondrial Haplogroup N9a Are Protected Against Metabolic Syndrome
Authors: Tanaka Masashi , Fuku Noriyuki , Nishigaki Yutaka , Matsuo Hitoshi , Segawa Tomonori , Watanabe Sachiro , Kato Kimihiko , Yoko Kiyoshi , Ito Masafumi , Nozawa Yoshinori , Yamada Yoshiji
Published: January 2007

To identify mitochondrial haplogroups that confer resistance against or susceptibility to metabolic syndrome, we performed a large-scale association study on 1,337 unrelated Japanese individuals, including 871 subjects with metabolic syndrome and 466 control subjects. Metabolic syndrome was diagnosed according to modified National Cholesterol Education Program Adult Treatment Panel III guidelines, using the cutoff point for obesity as a BMI of ≥25 kg/m2 instead of waist circumference. The genotypes for 25 polymorphisms in the coding region of the mitochondrial genome were determined, and the haplotypes were classified into 10 major haplogroups, i.e., F, B, A, N9a, M7a, M7b, G1, G2, D5, and D4. Multivariate logistic regression analysis revealed that the haplogroup N9a was significantly associated with resistance against metabolic syndrome in women with an odds ratio (OR) of 0.21 (95% CI 0.07–0.58, P = 0.0042). Women with haplogroups G1 and D5 tended to be resistant against metabolic syndrome with an OR of 0.22 (0.06–0.68, P = 0.0129) for G1 and with an OR of 0.32 (0.10–0.96, P = 0.0469) for D5, respectively. These results indicate that mitochondrial haplogroup N9a may be a protective factor against metabolic syndrome in Japanese women.
Treatment of Streptozotocin-Induced Diabetic Rats With AVE7688, a Vasopeptidase Inhibitor
Authors: Davidson Eric P. , Kleinschmidt Travis L. , Oltman Christine L. , Lund Donald D. , Yorek Mark A.
Published: January 2007

In epineurial arterioles, acetylcholine-mediated vascular relaxation is mediated by nitric oxide and endothelium-derived hyperpolarizing factor (EDHF), and both mechanisms are impaired by diabetes. The mediator responsible for the effect of EDHF is unknown. In epineurial arterioles, C-type natriuretic peptide (CNP) has properties consistent with EDHF-like activity. Epineurial arterioles express CNP, and exogenous CNP causes a concentration-dependent vascular relaxation. In streptozotocin-induced diabetic rats, CNP-mediated vascular relaxation in epineurial arterioles is decreased. Since CNP may be a regulator of vascular function, a vasopeptidase inhibitor may be an effective treatment for diabetes-induced vascular and neural disease. Vasopeptidase inhibitors inhibit ACE activity and neutral endopeptidase, which degrades natriuretic peptides. Streptozotocin-induced diabetic rats were treated with AVE7688 (450 mg/kg in the diet), a vasopeptidase inhibitor, for 8–10 weeks after 4 weeks of untreated diabetes. Treatment of diabetic rats corrected the diabetes-induced decrease in endoneurial blood flow, significantly improved motor and sensory nerve conduction velocity, prevented the development of hypoalgesia in the hind paw, and reduced superoxide and nitrotyrosine levels in epineurial arterioles. The diabetes-induced decrease in acetylcholine-mediated vascular relaxation by epineurial arterioles was significantly improved with treatment. These studies suggest that vasopeptidase inhibitors may be an effective approach for the treatment of diabetic vascular and neural dysfunction.
An ATP-Binding Mutation (G334D) in KCNJ11 Is Associated With a Sulfonylurea-Insensitive Form of Developmental Delay, Epilepsy, and Neonatal Diabetes
Authors: Masia Ricard , Koster Joseph C. , Tumini Stefano , Chiarelli Francesco , Colombo Carlo , Nichols Colin G. , Barbetti Fabrizio
Published: January 2007

Mutations in the pancreatic ATP-sensitive K+ channel (KATP channel) cause permanent neonatal diabetes mellitus (PNDM) in humans. All of the KATP channel mutations examined result in decreased ATP inhibition, which in turn is predicted to suppress insulin secretion. Here we describe a patient with severe PNDM, which includes developmental delay and epilepsy, in addition to neonatal diabetes (developmental delay, epilepsy, and neonatal diabetes [DEND]), due to a G334D mutation in the Kir6.2 subunit of KATP channel. The patient was wholly unresponsive to sulfonylurea therapy (up to 1.14 mg · kg−1 · day−1) and remained insulin dependent. Consistent with the putative role of G334 as an ATP-binding residue, reconstituted homomeric and mixed WT+G334D channels exhibit absent or reduced ATP sensitivity but normal gating behavior in the absence of ATP. In disagreement with the sulfonylurea insensitivity of the affected patient, the G334D mutation has no effect on the sulfonylurea inhibition of reconstituted channels in excised patches. However, in macroscopic rubidium-efflux assays in intact cells, reconstituted mutant channels do exhibit a decreased, but still present, sulfonylurea response. The results demonstrate that ATP-binding site mutations can indeed cause DEND and suggest the possibility that sulfonylurea insensitivity of such patients may be a secondary reflection of the presence of DEND rather than a simple reflection of the underlying molecular basis.
Wnt10b Inhibits Obesity in ob/ob and Agouti Mice
Authors: Wright Wendy S. , Longo Kenneth A. , Dolinsky Vernon W. , Gerin Isabelle , Kang Sona , Bennett Christina N. , Chiang Shian-Huey , Prestwich Tyler C. , Gress Catherine , Burant Charles F. , Susulic Vedrana S. , MacDougald Ormond A.
Published: January 2007

The Wnt family of secreted signaling molecules has profound effects on diverse developmental processes, including the fate of mesenchymal progenitors. While activation of Wnt signaling blocks adipogenesis, inhibition of endogenous Wnt/β-catenin signaling by Wnt10b promotes spontaneous preadipocyte differentiation. Transgenic mice with expression of Wnt10b from the FABP4 promoter (FABP4-Wnt10b) have less adipose tissue when maintained on a normal chow diet and are resistant to diet-induced obesity. Here we demonstrate that FABP4-Wnt10b mice largely avert weight gain and metabolic abnormalities associated with genetic obesity. FABP4-Wnt10b mice do not gain significant body weight on the ob/ob background, and at 8 weeks of age, they have an ∼70% reduction in visceral and subcutaneous adipose tissues compared with ob/ob mice. Similarly, on the lethal yellow agouti (Ay) background, FABP4-Wnt10b mice have 50–70% less adipose tissue weight and circulating leptin at 5 months of age. Wnt10b-Ay mice are more glucose tolerant and insulin sensitive than Ay controls, perhaps due to reduced expression and circulation of resistin. Reduced expression of inflammatory cytokines may also contribute to improved glucose homeostasis.
Myocardial Uptake of Circulating Triglycerides in Nondiabetic Patients With Heart Disease
Authors: Nelson Robert H. , Prasad Abhiram , Lerman Amir , Miles John M.
Published: January 2007

Animal studies indicate that oversupply of fatty acids derived from the action of cardiac lipoprotein lipase (LPL) on plasma lipoproteins may contribute to myocardial dysfunction. However, the contribution of circulating triglycerides to myocardial fatty acid supply in humans is not known. Six postabsorptive nondiabetic subjects who were scheduled for diagnostic coronary angiography were studied. 14C oleate and a lipid emulsion labeled with 3H triolein were infused to assess myocardial uptake of free fatty acids (FFAs) and triglycerides, as well as myocardial spillover of LPL-generated fatty acids. Six paired blood samples were taken from the femoral artery and the coronary sinus. Coronary sinus concentrations of unlabeled triglycerides were slightly, but not significantly, lower than arterial (P = 0.12), whereas labeled triglyceride concentrations were significantly lower in the coronary sinus than in the artery (P < 0.05; extraction fraction ≅11%). Triglycerides and FFAs accounted for ∼17% and ∼83%, respectively, of myocardial fatty acid uptake. Systemic and myocardial fractional spillover of LPL-generated fatty acids was 49.0 ± 7% and 34.7 ± 13%, respectively. The myocardium was a minor contributor to systemic triglyceride uptake (∼3%) and a trivial contributor to systemic FFA production (∼0.5%). These results indicate that circulating triglycerides may be a significant source of fatty acids for myocardial respiration.
Increasing Expenditure on Health Care Incurred by Diabetic Subjects in a Developing Country
Authors: Ramachandran Ambady , Ramachandran Shobhana , Snehalatha Chamukuttan , Augustine Christina , Murugesan Narayanasamy , Viswanathan Vijay , Kapur Anil , Williams Rhys
Published: January 2007

OBJECTIVE— This study aimed to assess the direct cost incurred by diabetic subjects who were in different income groups in urban and rural India, as well as to examine the changing trends of costs in the urban setting from 1998 to 2005.

RESEARCH DESIGN AND METHODS— A total of 556 diabetic subjects from various urban and rural regions of seven Indian states were enrolled. A brief uniform coded questionnaire (24 items) on direct cost was used.

RESULTS— Annual family income was higher in urban subjects (rupees [Rs] 100,000 or $2,273) than in the rural subjects (Rs 36,000 or $818) (P < 0.001). Total median expenditure on health care was Rs 10,000 ($227) in urban and Rs 6,260 ($142) in rural (P < 0.001) subjects. Treatment costs increased with duration of diabetes, presence of complications, hospitalization, surgery, insulin therapy, and urban setting. Lower-income groups spent a higher proportion of their income on diabetes care (urban poor 34% and rural poor 27%). After accounting for inflation, a secular increase of 113% was observed in the total expenses between 1998 and 2005 in the urban population. The highest increase in percentage of household income devoted to diabetes care was in the lowest economic group (34% of income in 1998 vs. 24.5% in 2005) (P < 0.01). There was a significant improvement in urban subjects in medical reimbursement from 2% (1998) to 21.3% (2005).

CONCLUSIONS— Urban and rural diabetic subjects spend a large percentage of income on diabetes management. The economic burden on urban families in developing countries is rising, and the total direct cost has doubled from 1998 to 2005.
Insulin Resistance as Estimated by Homeostasis Model Assessment Predicts Incident Symptomatic Cardiovascular Disease in Caucasian Subjects From the General Population
Authors: Bonora Enzo , Kiechl Stefan , Willeit Johann , Oberhollenzer Friedrich , Egger Georg , Meigs James B. , Bonadonna Riccardo C. , Muggeo Michele
Published: January 2007

OBJECTIVE—The purpose of this study was to evaluate whether insulin resistance is associated to cardiovascular disease (CVD) and to understand whether this association can be explained by traditional and novel CVD risk factors associated with this metabolic disorder.

RESEARCH DESIGN AND METHODS—We examined a sample representative of the population of Bruneck, Italy (n = 919; aged 40–79 years). Insulin-resistant subjects were those with a score in the top quartile of the homeostasis model assessment (HOMA) for insulin resistance (HOMA-IR). Risk factors correlated with insulin resistance included BMI, A1C, HDL cholesterol, triglycerides, blood pressure, high-sensitivity C-reactive protein (hsCRP), fibrinogen, oxidized LDL, vascular cell adhesion molecule-1 (VCAM-1), and adiponectin. Subjects without CVD at baseline were followed up for 15 years for incident CVD, a composite end point including fatal and nonfatal myocardial infarction and stroke, transient ischemic attack, and any revascularization procedure.

RESULTS—During follow-up, 118 subjects experienced a first symptomatic CVD event. Levels of HOMA-IR were higher at baseline among subjects who developed CVD (2.8) compared with those remaining free of CVD (2.5) (P < 0.05). Levels of HOMA-IR also were significantly correlated (P < 0.05) with most CVD risk factors we evaluated. In Cox proportional hazard models, insulin-resistant subjects had an age-, sex-, and smoking-adjusted 2.1-fold increased risk (95% CI 1.3–3.1) of incident symptomatic CVD relative to non–insulin-resistant subjects. After sequential adjustment for physical activity and classic risk factors (A1C, LDL cholesterol, and hypertension) as well as BMI, HDL cholesterol, triglycerides, and novel risk factors, including fibrinogen, oxidized LDL, hsCRP, VCAM-1, and adiponectin, the association between HOMA-IR and incident CVD remained significant and virtually unchanged (hazard ratio 2.2 [95% CI 1.4–3.6], P < 0.001).

CONCLUSIONS—HOMA-estimated insulin resistance is associated with subsequent symptomatic CVD in the general population independently of all classic and several nontraditional risk factors. These data suggest that insulin resistance may be an important target to reduce CVD risk.
Effect of Adjunctive Pramlintide Treatment on Treatment Satisfaction in Patients With Type 1 Diabetes
Authors: Marrero David G. , Crean John , Zhang Bei , Kellmeyer Terrie , Gloster Maurice , Herrmann Kathrin , Rubin Richard , Fineberg Naomi , Kolterman Orville
Published: January 2007

OBJECTIVE—To assess the effect of adjunctive pramlintide treatment on treatment satisfaction in patients with type 1 diabetes treated with intensive insulin regimens.

RESEARCH DESIGN AND METHODS—Intensively treated (multiple daily injection [MDI] or continuous subcutaneous insulin infusion [CSII] pump therapy) patients with type 1 diabetes completed a study-specific treatment satisfaction questionnaire following 29 weeks of either placebo (n = 136) or pramlintide (n = 130) treatment in a double-blind, noninferiority pramlintide dose titration trial. End points included patient reported outcomes, their relationship to insulin treatment regimen, A1C, weight, and insulin use.

RESULTS—Pramlintide-treated patients reported greater treatment satisfaction in most questionnaire responses. Treatment satisfaction was similar for pramlintide-treated patients regardless of intensive insulin regimens (MDI versus CSII). Mean A1C was reduced to a similar degree in both pramlintide- (−0.39 ± 0.07%) and placebo-treated (−0.45 ± 0.07%) patients. However, pramlintide treatment was associated with reductions in mean body weight (−1.50 ± 0.33 kg; P < 0.0001) and mealtime insulin use (−19.05 ± 5.17%; P < 0.005) over 29 weeks, while placebo treatment resulted in weight gain (1.28 ± 0.25 kg) and a smaller reduction in mealtime insulin use (−2.20 ± 3.33%).

CONCLUSIONS—Despite similar reductions in A1C, pramlintide treatment resulted in greater treatment satisfaction compared with placebo treatment. This was independent of insulin delivery method.
Ethnicity, Insulin Resistance, and Inflammatory Adipokines in Women at High and Low Risk for Vascular Disease
Authors: Silha Josef V. , Nyomba B.L. Grégoire , Leslie William D. , Murphy Liam J.
Published: January 2007

OBJECTIVE—We sought to compare the relationship between body composition, insulin resistance, and inflammatory adipokines in Aboriginal Canadian women, who are at high risk of vascular disease, with white women.

RESEARCH DESIGN AND METHODS—A subgroup of the First Nations Bone Health Study population, consisting of 131 Aboriginal women and 132 matched white women, was utilized. Body composition was determined by whole-body dual X-ray absorptiometry, and blood analytes were measured after an overnight fast.

RESULTS—After excluding individuals with diabetes, A1C, BMI, percent trunk fat, and homeostasis model assessment of insulin resistance (HOMA-IR) were greater in First Nation women compared with white women, whereas adiponectin, retinol binding protein (RBP)4, and insulin-like growth factor binding protein-1 (IGFBP-1) were lower. First Nation women had more trunk fat for any given level of total fat than white women. There were no differences in resistin, leptin, tumor necrosis factor (TNF)-α, or C-reactive protein (CRP) levels between First Nation and white women. Insulin resistance correlated with leptin and inversely with adiponectin levels in both First Nation and white women. There were weak correlations between insulin resistance and TNF-α, interleukin-6, and CRP, but these were not significant after correction for body fat. No correlation was found between RBP4 and insulin resistance. ANCOVA revealed a higher HOMA-IR adjusted for total body fat in First Nation women than in white women (P = 0.015) but not HOMA-IR adjusted for trunk fat (P > 0.2).

CONCLUSIONS—First Nation women are more insulin resistant than white women, and this is explained by trunk fat but not total fat. Despite the increased insulin resistance, inflammatory adipokines are not significantly increased in First Nation women compared with white women.
High Risk of Cardiovascular Mortality in Individuals With Impaired Fasting Glucose Is Explained by Conversion to Diabetes
Authors: Rijkelijkhuizen Josina M. , Nijpels Giel , Heine Robert J. , Bouter Lex M. , Stehouwer Coen D.A. , Dekker Jacqueline M.
Published: January 2007

OBJECTIVE—To optimize identification of future diabetic patients, the American Diabetes Association (ADA) introduced criteria for impaired fasting glucose (IFG) in 1997 (IFG 6.1 mmol/l [IFG6.1]) and lowered the threshold from 6.1 to 5.6 mmol/l (IFG5.6) in 2003. Our aim was to assess the consequences of lowering the IFG cutoff on the risk of cardiovascular disease (CVD) mortality and to evaluate whether this risk is explained by a conversion to type 2 diabetes within 6.4 years.

RESEARCH DESIGN AND METHODS—In a population-based cohort, the Hoorn Study, plasma glucose was determined in 1989 and 1996 (n = 1,428). Subjects were classified in 1989 according to 1997 and 2003 ADA criteria. Subjects with IFG in 1989 were further classified according to diabetes status in 1996. Hazard ratios for CVD mortality (n = 81) in the period 1996–2005 were adjusted for age and sex.

RESULTS—Subjects with IFG6.1, but not IFG5.6, had a significantly higher CVD mortality risk than normal fasting glucose (NFG) subjects. Subjects who converted from IFG to diabetes (IFG6.1: 42%; IFG5.6: 21%) had a more than twofold risk of CVD mortality (IFG6.1: 2.47 [1.17–5.19]; IFG5.6: 2.14 [1.12–4.10]) than subjects with NFG. IFG subjects who did not develop diabetes did not have significantly higher CVD mortality risks (IFG6.1: 1.50 [0.72–3.15]; IFG5.6: 1.15 [0.69–1.93]).

CONCLUSIONS—The lower cutoff for IFG (ADA 2003 criteria) results in a category of IFG that no longer represents a high-risk state of CVD. Furthermore, only subjects who convert from IFG to diabetes have a high risk of CVD mortality.
The BIGTT Test
Authors: Hansen Torben , Drivsholm Thomas , Urhammer Søren A. , Palacios Rene T. , Vølund Aage , Borch-Johnsen Knut , Pedersen Oluf
Published: January 2007

OBJECTIVE—Insulin resistance and impaired β-cell function are key elements in the pathogenesis of type 2 diabetes. We aimed to develop valid algorithms for estimation of the insulin sensitivity index (SI) and acute insulin response (AIR) derived from simple and cheap physiological measurements that could be used in large-scale metabolic, genetic, and epidemiological studies.

RESEARCH DESIGN AND METHODS—For our purpose, data from an oral glucose tolerance test (OGTT) (18 samples during 240 min) and a tolbutamide-modified intravenous glucose tolerance test (IVGTT) (33 samples during 180 min) from 258 individuals with fasting plasma glucose <7 mmol/l and 2-h plasma glucose <7.8 mmol/l were used for model development and internal validation. Data from an additional 28 individuals were used for external validation. Bergman’s minimal model was used to calculate SI, and the trapezoidal method was used to calculate AIR0–8 min. Multiple linear regression was applied to derive predictive equations of log(SI) and log(AIR0–8 min) using data on sex, BMI, plasma glucose, and serum insulin levels obtained during the OGTT.

RESULTS—We demonstrate that it is possible to obtain estimates of SI (BIGTT-SI) and AIR (BIGTT-AIR) that are highly correlated to IVGTT-derived values of SI (R2 = 0.77) and AIR (R2 = 0.54). In the two validation datasets we obtained similar results.

CONCLUSIONS—Data from OGTTs can provide accurate measures of insulin sensitivity and β-cell function, which can be used in large scale metabolic, genetic, and epidemiological studies.
Functional Vascular Endothelial Growth Factor −634G>C SNP Is Associated With Proliferative Diabetic Retinopathy
Authors: Errera Flavia I.V. , Canani Luís Henrique , Silva Maria Elisabeth R. , Yeh Erika , Takahashi Walter , Santos Katia G. , Souto Katia E.P. , Tschiedel Balduíno , Roisenberg Israel , Gross Jorge Luis , Passos-Bueno Maria Rita
Published: January 2007

OBJECTIVE—The purpose of this study was to evaluate the effect of the single nucleotide polymorphism (SNP) −634G>C at the 5′ regulatory region of the vascular endothelial growth factor (VEGF) in the risk of proliferative diabetic retinopathy (PDR) in the Brazilian population of European ancestry with type 2 diabetes.

RESEARCH DESIGN AND METHODS—A case-control study was conducted in 501 type 2 diabetic patients of European ancestry. Patients underwent a standardized clinical, ophthalmological, and laboratory evaluation. Of these, 167 patients had PDR (case patients), and 334 were considered as control subjects (patients without PDR) for PDR. A reference population (110 individuals of European ancestry) was also evaluated.

RESULTS—No evidence of association between −634G>C/VEGF and the presence of diabetic retinopathy or type 2 diabetes was observed (P > 0.05). However, CC homozygous for the SNP −634G>C was significantly more frequent in patients with PDR (37 of 167; 22.2%) than in the corresponding control group (40 of 334; 12%) in accordance with a recessive model (P = 0.003). This effect was further observed when creatinine, BMI, sex, duration of type 2 diabetes, HDL cholesterol, and systolic blood pressure were taken into account (odds ratio 1.9 [95% CI 1.01–3.79], P = 0.04).

CONCLUSIONS—The presence of the allele −634C/VEGF in homozygosity is an independent risk factor for the development of PDR in type 2 diabetic patients of European ancestry.
Cardiometabolic Risk in Impaired Fasting Glucose and Impaired Glucose Tolerance
Authors: Pankow James S. , Kwan David K. , Duncan Bruce B. , Schmidt Maria I. , Couper David J. , Golden Sherita , Ballantyne Christie M.
Published: January 2007

OBJECTIVE—We compared and contrasted cardiovascular disease (CVD) risk factors, subclinical manifestations of CVD, incident coronary heart disease (CHD), and all-cause mortality by categories of impaired glucose regulation in nondiabetic individuals.

RESEARCH DESIGN AND METHODS—The study included 6,888 participants aged 52–75 years who had no history of diabetes or CVD. All-cause mortality and incident CHD were ascertained over a median of 6.3 years of follow-up.

RESULTS—Agreement between fasting and postchallenge glucose impairment was poor: 3,048 subjects (44%) had neither impaired fasting glucose (IFG) nor impaired glucose tolerance (IGT), 1,690 (25%) had isolated IFG, 1,000 (14%) had isolated IGT, and 1,149 (17%) had both IFG and IGT. After adjustment for age, sex, race, and center, subjects with isolated IFG were more likely to smoke, consume alcohol, and had higher mean BMI, waist circumference, LDL cholesterol, and fasting insulin and lower HDL cholesterol than those with isolated IGT, while subjects with isolated IGT had higher mean triglycerides, systolic blood pressure, and white cell counts. Measures of subclinical CVD and rates of all-cause mortality and incident CHD were similar in isolated IFG and isolated IGT.

CONCLUSIONS—Neither isolated IFG nor isolated IGT was associated with a more adverse CVD risk profile.
Prevalence of Candida glabrata and Its Response to Boric Acid Vaginal Suppositories in Comparison With Oral Fluconazole in Patients With Diabetes and Vulvovaginal Candidiasis
Authors: Ray Debarti , Goswami Ravinder , Banerjee Uma , Dadhwal Vatsla , Goswami Deepti , Mandal Piyali , Sreenivas Vishnubhatla , Kochupillai Narayana
Published: January 2007

OBJECTIVE—A large proportion of vulvovaginal candidiasis (VVC) in diabetes is due to non–albicans Candida species such as C. glabrata and C. tropicalis. Observational studies indicate that diabetic patients with C. glabrata VVC respond poorly to azole drugs. We evaluated the response to oral fluconazole and boric acid vaginal suppositories in diabetic patients with VVC.

RESEARCH DESIGN AND METHODS—A total of 112 consecutive diabetic patients with VVC were block randomized to receive either single-dose oral 150-mg fluconazole or boric acid vaginal suppositories (600 mg/day for 14 days). The primary efficacy outcome was the mycological cure in patients with C. glabrata VVC in the two treatment arms. The secondary outcomes were the mycological cure in C. albicans VVC, overall mycological cure irrespective of the type of Candida species, frequencies of yeast on direct microscopy, and clinical symptoms and signs of VVC on the 15th day of treatment. Intention-to-treat (ITT; n = 111) and per-protocol (PP; n = 99) analyses were performed.

RESULTS—C. glabrata was isolated in 68 (61.3%) and C. albicans in 32 (28.8%) of 111 subjects. Patients with C. glabrata VVC showed higher mycological cure with boric acid compared with fluconazole in the ITT (21 of 33, 63.6% vs. 10 of 35, 28.6%; P = 0.01) and PP analyses (21 of 29, 72.4% vs. 10 of 30, 33.3%; P = 0.01). The secondary efficacy outcomes were not significantly different in the two treatment arms in the ITT and PP analyses.

CONCLUSIONS—Diabetic women with C. glabrata VVC show higher mycological cure with boric acid vaginal suppositories given for 14 days in comparison with single-dose oral 150-mg fluconazole.
Androgens and Diabetes in Men
Authors: Selvin Elizabeth , Feinleib Manning , Zhang Lei , Rohrmann Sabine , Rifai Nader , Nelson William G. , Dobs Adrian , Basaria Shehzad , Golden Sherita Hill , Platz Elizabeth A.
Published: January 2007

OBJECTIVE—Low levels of androgens in men may play a role in the development of diabetes; however, few studies have examined the association between androgen concentration and diabetes in men in the general population. The objective of this study is to test the hypothesis that low normal levels of total, free, and bioavailable testosterone are associated with prevalent diabetes in men.

RESEARCH DESIGN AND METHODS—The study sample included 1,413 adult men aged ≥20 years who participated in the morning session of the first phase of the Third National Health and Nutrition Examination Survey, a cross-sectional survey of the civilian, noninstitutionalized population of the U.S. Bioavailable and free testosterone levels were calculated from serum total testosterone, sex hormone–binding globulin, and albumin concentrations.

RESULTS—In multivariable models adjusted for age, race/ethnicity, and adiposity, men in the first tertile (lowest) of free testosterone level were four times more likely to have prevalent diabetes compared with men in the third tertile (odds ratio 4.12 [95% CI 1.25–13.55]). Similarly, men in the first tertile of bioavailable testosterone also were approximately four times as likely to have prevalent diabetes compared wth men in the third tertile (3.93 [1.39–11.13]). These associations persisted even after excluding men with clinically abnormal testosterone concentrations defined as total testosterone <3.25 ng/ml or free testosterone <0.07 ng/ml. No clear association was observed for total testosterone after multivariable adjustment (P for trend across tertiles = 0.27).

CONCLUSIONS—Low free and bioavailable testosterone concentrations in the normal range were associated with diabetes, independent of adiposity. These data suggest that low androgen levels may be a risk factor for diabetes in men.
How Reliable Is Estimation of Glomerular Filtration Rate at Diagnosis of Type 2 Diabetes?
Authors: Chudleigh Richard A. , Dunseath Gareth , Evans William , Harvey John N. , Evans Philip , Ollerton Richard , Owens David R.
Published: January 2007

OBJECTIVE—The Cockcroft-Gault (CG) and Modification of Diet in Renal Disease (MDRD) equations previously have been recommended to estimate glomerular filtration rate (GFR). We compared both estimates with true GFR, measured by the isotopic 51Cr-EDTA method, in newly diagnosed, treatment-naïve subjects with type 2 diabetes.

RESEARCH DESIGN AND METHODS—A total of 292 mainly normoalbuminuric (241 of 292) subjects were recruited. Subjects were classified as having mild renal impairment (group 1, GFR <90 ml/min per 1.73 m2) or normal renal function (group 2, GFR ≥90 ml/min per 1.73 m2). Estimated GFR (eGFR) was calculated by the CG and MDRD equations. Blood samples drawn at 44, 120, 180, and 240 min after administration of 1 MBq of 51Cr-EDTA were used to measure isotopic GFR (iGFR).

RESULTS—For subjects in group 1, mean (±SD) iGFR was 83.8 ± 4.3 ml/min per 1.73 m2. eGFR was 78.0 ± 16.5 or 73.7 ± 12.0 ml/min per 1.73 m2 using CG and MDRD equations, respectively. Ninety-five percent CIs for method bias were –11.1 to −0.6 using CG and –14.4 to –7.0 using MDRD. Ninety-five percent limits of agreement (mean bias ± 2 SD) were −37.2 to 25.6 and −33.1 to 11.7, respectively. In group 2, iGFR was 119.4 ± 20.3 ml/min per 1.73 m2. eGFR was 104.4 ± 26.3 or 92.3 ± 18.7 ml/min per 1.73 m2 using CG and MDRD equations, respectively. Ninety-five percent CIs for method bias were –17.4 to –12.5 using CG and –29.1 to –25.1 using MDRD. Ninety-five percent limits of agreement were −54.4 to 24.4 and −59.5 to 5.3, respectively.

CONCLUSIONS—In newly diagnosed type 2 diabetic patients, particularly those with a GFR ≥90 ml/min per 1.73 m2, both CG and MDRD equations significantly underestimate iGFR. This highlights a limitation in the use of eGFR in the majority of diabetic subjects outside the setting of chronic kidney disease.
Sex Differences in Endothelial Function Markers Before Conversion to Pre-Diabetes: Does the Clock Start Ticking Earlier Among Women?
Authors: Donahue Richard P. , Rejman Karol , Rafalson Lisa B. , Dmochowski Jacek , Stranges Saverio , Trevisan Maurizio
Published: January 2007

OBJECTIVE—We examined whether biomarkers of endothelial function, fibrinolysis/thrombosis and adiponectin, predict the progression from normal to pre-diabetes more strongly among women than men over 6 years of follow-up from the Western New York Health Study.

RESEARCH DESIGN AND METHODS—In 2002–2004, 1,455 participants from the Western New York Health Study, who were free of type 2 diabetes and cardiovascular disease at baseline (1996–2001), were selected for reexamination. An incident case of pre-diabetes was defined as fasting glucose <100 mg/dl at the baseline examination and ≥100 and <126 mg/dl at the follow-up examination. Biomarkers of endothelial function (E-selectin and soluble intracellular adhesion molecule-1 [sICAM-1]), fibrinolysis/thrombosis (plasminogen activator inhibitor-1 [PAI-1]), and fasting insulin, adiponectin, and inflammation (high-sensitivity C-reactive protein) were measured in frozen (−190°C) baseline samples.

RESULTS—Multivariate analyses revealed higher adjusted mean values of biomarkers of endothelial dysfunction (E-selectin and sICAM-1) and fibrinolysis (PAI-1) and lower mean values of adiponectin only among women who developed pre-diabetes compared with control subjects. Formal tests for interaction between sex and case/control status were statistically significant for E-selectin (P = 0.042), PAI-1 (P = 0.001), sICAM-1 (P = 0.011), and frequency of hypertension (P < 0.001).

CONCLUSIONS—These results support the concept that women who progressed from normoglycemia to pre-diabetes have greater endothelial dysfunction than men as well as more hypertension and a greater degree of fibrinolysis/thrombosis. Whether this relates to the higher risk of heart disease among diabetic women awaits further study.
A Systematic Review and Meta-Analysis of Hypoglycemia and Cardiovascular Events
Authors: Gangji Azim S. , Cukierman Tali , Gerstein Hertzel C. , Goldsmith Charles H. , Clase Catherine M.
Published: January 2007

OBJECTIVE—Glyburide is the most widely used sulfonylurea but has unique pharmacodynamic properties that may increase harm. We hypothesized that glyburide causes more hypoglycemia and cardiovascular events than other secretagogues or insulin.

RESEARCH DESIGN AND METHODS—Data sources were Medline, Embase, Cochrane, and three other web-based clinical trial registers (1966–2005). Parallel, randomized, controlled trials in people with type 2 diabetes comparing glyburide monotherapy with monotherapy using secretagogues or insulin were selected. Outcomes were hypoglycemia, glycemic control, cardiovascular events, body weight, and death. Titles and abstracts of 1,806 publications were reviewed in duplicate and 21 relevant articles identified. Data on patient characteristics, interventions, outcomes, and validity were extracted in duplicate using predefined criteria.

RESULTS—Glyburide was associated with a 52% greater risk of experiencing at least one episode of hypoglycemia compared with other secretagogues (relative risk 1.52 [95% CI 1.21–1.92]) and with 83% greater risk compared with other sulfonylureas (1.83 [1.35–2.49]). Glyburide was not associated with an increased risk of cardiovascular events (0.84 [0.56–1.26]), death (0.87 [0.70–1.07]), or end-of-trial weight (weighted mean difference 1.69 kg [95% CI −0.41 to 3.80]) compared with other secretagogues. Limitations included suboptimal reporting of original trials. Loss to follow-up exceeded 20% in some studies, and major hypoglycemia was infrequently reported.

CONCLUSIONS—Glyburide caused more hypoglycemia than other secretagogues and other sulfonylureas. Glyburide was not associated with an increased risk of cardiovascular events, death, or weight gain.
Physical Activity and Metabolic Risk in Individuals With a Family History of Type 2 Diabetes
Authors: Ekelund Ulf , Griffin Simon J. , Wareham Nicholas J. ,   on behalf of the ProActive Research Group
Published: January 2007

OBJECTIVE—We sought to examine the independent associations between different dimensions of physical activity with intermediary and clustered metabolic risk factors in overweight individuals with an increased risk of type 2 diabetes to inform future preventive action.

RESEARCH DESIGN AND METHODS—We measured total body movement and five other subcomponents of physical activity by accelerometry in 258 adults (aged 30–50 years) with a family history of type 2 diabetes. We estimated aerobic fitness from an incremental treadmill exercise test. We measured body composition by bioimpedance and waist circumference, blood pressure, fasting triglycerides, HDL cholesterol, glucose, and insulin with standard methods. We constructed a standardized continuously distributed variable for clustered risk.

RESULTS—Total body movement (counts · day−1) was significantly and independently associated with three of six risk factors (fasting triglycerides, insulin, and HDL) and with clustered metabolic risk (P = 0.004) after adjustment for age, sex, and obesity. Time spent at moderate- and vigorous-intensity physical activity (MPVA) was independently associated with clustered metabolic risk (P = 0.03). Five- and 10-min bouts of MVPA, time spent sedentary, time spent at light-intensity activity, and aerobic fitness were not significantly related with clustered risk after adjustment for confounding factors.

CONCLUSIONS—Total body movement is associated with intermediary phenotypic risk factors for cardiovascular disease and metabolic disease and with clustered metabolic risk independent of aerobic fitness and obesity. Increasing the total amount of physical activity in sedentary and overweight individuals may have beneficial effects on metabolic risk factors.
Effect of a Nurse-Directed Diabetes Disease Management Program on Urgent Care/Emergency Room Visits and Hospitalizations in a Minority Population
Authors: Davidson Mayer B. , Ansari Adeela , Karlan Vicki J.
Published: January 2007

OBJECTIVE—To evaluate whether nurse-directed diabetes care reduced preventable diabetes-related urgent care/emergency room visits and hospitalizations in a minority population.

RESEARCH DESIGN AND METHODS—Diabetic patients who receive care in a county public health clinic were randomly selected for a Diabetes Managed Care Program (DMCP) in which a specially trained nurse followed detailed treatment algorithms to provide diabetes care for 1 year. Preventable diabetes-related urgent care/emergency room visits and hospitalizations for these patients incurred during the intervention year and the year before enrollment were compared. Preventable diabetes-related causes were defined as metabolic (diabetic ketoacidosis, hyperglycemia, or hypoglycemia) or infection (cellulitis, foot ulcer, osteomyelitis, fungal infection, or urinary tract infection).

RESULTS—Use of the urgent care/emergency room and hospitalizations during the intervention year and the year prior were available for 331 patients who completed the DMCP intervention. There were 94 total urgent care/emergency room visits and hospitalizations in the year before entering the DMCP and 46 during the DMCP year, a 51% reduction. Preventable diabetes-related episodes were far fewer. During the prior year, 14 patients made 15 urgent care/emergency room visits and 5 patients incurred 6 hospitalizations. During the DMCP year, four different patients made five emergency room/urgent care visits and one other patient was hospitalized. Preventable diabetes-related use was significantly (P < 0.001) lower during the intervention year compared with the prior year. Total charges for urgent care/emergency room visits and hospitalizations only (not other charges related to diabetes care) during the year before entering the DMCP were $129,176 compared with $24,630 during the DMCP year.

CONCLUSIONS—When compared with usual care, nurse-directed diabetes care resulted in significantly fewer urgent care/emergency room visits and hospitalizations for preventable diabetes-related causes. Policy makers seeking to improve diabetes care and conserve resources should seriously consider adopting this approach.
Whole‐Cell Recording In Vivo
Authors: R. DeWeese Michael
Published: January 2007

In vivo whole‐cell patch‐clamp recording provides a means for measuring membrane currents and potentials from individual cells in the intact animal. Patch‐clamp methods have largely been developed in vitro. This body of work has contributed enormously to the understanding of many important phenomena in excitable cells—including synaptic plasticity in the mammalian central nervous system, and the behavior of individual protein channels. In recent years, an increasing number of groups have applied whole‐cell recording techniques in the intact animal. Such in vivo studies offer the tantalizing possibility of uncovering the underlying principles and mechanisms of neural interactions within the natural context of fully intact biological networks. This unit focuses on strategies for overcoming the specific technical challenges posed by in vivo whole‐cell recording. A straightforward procedure is described for obtaining whole‐cell records from the cortex of the anesthetized rat; this procedure has also been applied successfully to awake animals and other rodent species with minor modifications.
Isolation of Phosphopeptides by Immobilized Metal Ion Affinity Chromatography
Authors: Nühse Thomas , Yu Kebing , Salomon Arthur
Published: January 2007

The identification of protein phosphorylation sites from cell‐derived proteins is crucial to the understanding of signal transduction pathways. While determining the modified sites on individual proteins can present a significant challenge, recent progress in the rapid, large‐scale identification of phosphopeptides from complex protein mixtures by combinations of affinity chromatography and mass spectrometry provides a powerful tool to decipher the phosphoproteome. A set of protocols is described for sample preparation, fractionation, immobilized metal ion affinity chromatography (IMAC), and mass spectrometric analysis of phosphorylation sites. Parts of the protocols can be combined in different ways to adapt to sample amounts, complexity, and available equipment. Up to thousands of unique phosphopeptides can be sequenced from a single sample in a day, revealing a unique snapshot of global cellular phosphorylation sites on proteins and facilitating in‐depth study of the identified phosphoproteins.
Human Artificial Chromosome Assembly by Transposon‐Based Retrofitting of Genomic BACs with Synthetic Alpha‐Satellite Arrays
Authors: Basu Joydeep , Willard Huntington F. , Stromberg Gregory
Published: January 2007

The development of methodologies for the rapid assembly of synthetic alpha‐satellite arrays recapitulating the higher‐order periodic organization of native human centromeres permits the systematic investigation of the significance of primary sequence and sequence organization in centromere function. Synthetic arrays with defined mutations affecting sequence and/or organization may be evaluated in a de novo human artificial chromosome assay. This unit describes strategies for the assembly of custom built alpha‐satellite arrays containing any desired mutation as well as strategies for the construction and manipulation of alpha satellite–based transposons. Transposons permit the rapid and reliable retrofitting of any genomic bacterial artificial chromosome (BAC) with synthetic alpha‐satellite arrays and other functional components, thereby facilitating conversion into BAC‐based human artificial chromosome vectors. These techniques permit identification and optimization of the critical parameters underlying the unique ability of alpha‐satellite DNA to facilitate de novo centromere assembly, and they will establish the foundation for the next generation of human artificial chromosome vectors.
Production of Vesicular Stomatitis Virus G Glycoprotein (VSV‐G) Pseudotyped Retroviral Vectors
Authors: Lo Hsin‐Lung , Yee Jiing‐Kuan
Published: January 2007

Retrovirus pseudotype is defined as the genome of one retrovirus encapsidated by the envelope protein of a second virus. The host range of the pseudotype is that of the virus donating the envelope protein. Two procedures that use 293GP cells, which are derived from human kidney 293 cells, are described here. The first is based on the high transient transfection efficiency of 293 cells. The retroviral construct and an expression plasmid for VSV‐G are co‐transfected into 293GP cells that stably express MLV gag and pol proteins. Transiently generated virus is then harvested during consecutive days following DNA transfection. The second procedure involves stable 293GP cell lines containing the VSV‐G gene under the control of a promoter whose activity is regulated by tetracycline. Cell lines containing the retroviral vector of interest are established under noninduced conditions. Infectious virus can be harvested following the induction of VSV‐G expression in these cell lines.
Flow Cytometry of the Side Population (SP)
Authors: Petriz Jordi
Published: January 2007

The side population (SP) has become an important hallmark for the definition of the stem‐cell compartment, especially for the detection of stem cells and for their physical isolation by fluorescence‐activated cell sorting (FACS). SP cells are CD34− and were discovered using ultraviolet excitation based on the efflux of Hoechst 33342 (Ho342). Although the method works as originally described, the protocol is difficult for most investigators to perform: first, because the ability to discriminate SP cells is based on the differential retention of Ho342 during a functional assay; second, because of the difficulties in setting the right experimental and acquisition conditions; and third, because analysis of the acquired data requires extensive expertise in flow cytometry to accurately detect the SP events. This unit is packed with helpful pointers to aid the user in obtaining the best possible results.
Validation of Tandem Mass Spectrometry Database Search Results Using DTASelect
Authors: Cociorva Daniel , L. Tabb David , Yates John R.
Published: January 2007

DTASelect provides a means by which complex SEQUEST results can be filtered, organized, and viewed. A single sample may produce tens of thousands of tandem mass spectra. Manually perusing and selecting SEQUEST matches among such a mass of data carries a risk of inconsistency. DTASelect allows the user to set complex criteria for acceptance or rejection of individual spectrum results. It also features rules for dealing with multiple, identical peptide matches and for removing proteins that are insufficiently evidenced. It provides its sorted and filtered summary as HTML and text documents for easy review and also offers several auxiliary reports. DTASelect is a powerful tool for automatic analysis of complex mixture tandem mass spectrometry.
Unix Survival Guide
Authors: Stein Lincoln D.
Published: January 2007

For a mixture of historical and practical reasons, much of the bioinformatics software discussed in this series runs on Linux, Mac OSX, Solaris, or one of the many other Unix variants. This appendix provides the novice with easy‐to‐understand information needed to survive in the Unix environment.