Maternal and fetal leptin,adiponectin levels and associations with fetal insulin sensitivity

Objective:

It remains uncertain whether leptin and adiponectin levels are correlated in maternal vs. fetal circulations. Little is known about whether leptin and adiponectin affect insulin sensitivity during fetal life.

Design and Methods:

In a prospective singleton pregnancy cohort (n = 248), we investigated leptin and adiponectin concentrations in maternal (at 24‐28 and 32‐35 weeks of gestation) and fetal circulations, and their associations with fetal insulin sensitivity (glucose/insulin ratio, proinsulin level).

Results:

Comparing concentrations in cord vs. maternal blood, leptin levels were 50% lower, but adiponectin levels more than doubled. Adjusting for gestational age at blood sampling, consistent and similar positive correlations (correlation coefficients: 0.31‐0.34, all P < 0.0001) were observed in leptin or adiponectin levels in maternal (at 24‐28 or 32‐25 weeks of gestation) vs. fetal circulations. For each SD increase in maternal plasma concentration at 24‐28 weeks, cord plasma concentration increased by 12.7 (95% confidence interval 6.8‐18.5) ng/ml for leptin, and 2.9 (1.8‐4.0) µg/ml for adiponectin, respectively (adjusted P < 0.0001). Fetal insulin sensitivity was negatively associated with cord blood leptin (each SD increase was associated with a 5.4 (2.1‐8.7) mg/dl/µU/ml reduction in cord plasma glucose/insulin ratio, and a 5.6 (3.9, 7.4) pmol/l increase in proinsulin level, all adjusted P < 0.01) but not adiponectin (P > 0.4) levels). Similar associations were observed in nondiabetic full‐term pregnancies (n = 211).

Conclusions:

The results consistently suggest a maternal impact on fetal leptin and adiponectin levels, which may be an early life pathway in maternal‐fetal transmission of the propensity to obesity and insulin resistance.     

Are Elevated Levels of IGF-1 Caused by Coronary Arteriesoclerosis?: Molecular and Clinical Analysis

The importance of insulin-like growth factor-1 (IGF-1) in coronary artery disease (CAD) due to wide range of its biological effects and its therapeutic potential, has already been described. Our aim was to evaluate possible influence of IGF-1 serum level changes on coronary atherosclerosis. In case of existence of such association our further aim was to verify and explain this phenomenon by examination of promoter P1 of IGF-1gene and receptor gene for IGF-1. The study was performed in 101 consecutive patients undergo for routine coronary angiography. Quantitative and qualitative assessment of coronary atherosclerosis was performed respectively by estimation of the number of culprit lesions in coronary arteries and by Gensini score calculation. IGF-1, IGFBP3 and plasma lipoproteins were measured in all patients. In addition, we evaluated DNA from 101 patients, isolated from blood cells, which was amplified by using PCR with sophisticated primers for P1 promoter of IGF-1 gene and IGF-1 receptor gene, then analyzed utilizing SSCP technique and automatically sequenced. We observed significant increase of serum IGF-1 levels in patients with “3 vessel disease” and with high score in Gensini scale when compared to those without any narrowing lesions in coronary arteries and 0 Gensini score (in group with 3 vessel disease 215.0 ± 71.3 versuss 176.7 ± 34.2 ng/ml p = 0.04 and with high Gensini score 231.4 ± 59.3 versus 181.0 ± 37.8 ng/ml p = 0.01).We found different genotypes for five P1 promoter polymorphisms of IGF-1 gene (RS35767, RS5742612, RS228837, RS11829693, RS17879774). There were no significant associations between the observed single nucleotide polymorphism (SNP) and coronary atherosclerosis nor with levels of circulating IGF-1. We found no structural polymorphism in receptor gene for IGF-1 nor in its extracellular domain(exon 2–4) nor in internal domain (exon 16–21). The effect of increased IGF-1 serum level in our study was probably independent from structural polymorphism in promoter P1 for IGF-1 or in receptor gene for IGF-1.     

Differential Effect of Weight Loss on Adipocyte Size Subfractions in Patients With Type 2 Diabetes

The size of adipocytes influences their function suggesting a differential responsiveness to intervention. We hypothesized that weight loss in patients with type 2 diabetes mellitus (T2DM) predominantly decreases the size of large and very‐large adipocyte subfractions in parallel with beneficial changes in serum adipokines and improved insulin sensitivity. A total of 44 volunteers from the Look Action for Health in Diabetes trial, who lost weight after 1‐year of intense lifestyle intervention, were included. Insulin sensitivity (hyperinsulinemic–euglycemic clamp), size of subcutaneous abdominal adipocytes (osmium fixation), and selected serum adipokines were measured. A 13% weight loss was accompanied by 46% improvement in insulin sensitivity (increased glucose disposal rate from 5.9 ± 2.2 to 8.6 ± 2.7 mg/min/kg fat‐free mass, P < 0.05) in parallel with a 36% increase in plasma adiponectin concentration (6.1 ± 3.1 to 8.3 ± 3.9 µg/ml, P < 0.05], but no changes in the proinflammatory cytokines interleukin‐6 and tumor necrosis factor‐α. Change in adiponectin correlated with changes in glucose disposal rate (r = 0.34, P < 0.05). Mean adipocyte size decreased (0.84 ± 0.25 to 0.64 ± 0.23 µl, P < 0.05), mainly due to changes in the large adipocyte subfraction (size 0.75–0.44 µl, relative number 19–26%; P < 0.05). Our data suggest that change in the large adipocyte subfraction may contribute to the improvement in insulin sensitivity via an increase in serum adiponectin. Such a relationship, which does not imply cause and effect, could not be obtained by measuring only mean adipocyte size. These data provide support for the measures of adipocyte size distribution in concert with in vitro adipokine secretion and lipolysis in future studies.     

Hydroxychloroquine decreases human MSC‐derived osteoblast differentiation and mineralization in vitro

We recently showed that patients with primary Sjögren Syndrome (pSS) have significantly higher bone mineral density (BMD) compared to healthy controls. The majority of those patients (69%) was using hydroxychloroquine (HCQ), which may have favourable effects on BMD. To study the direct effects of HCQ on human MSC‐derived osteoblast activity. Osteoblasts were cultured from human mesenchymal stromal cells (hMSCs). Cultures were treated with different HCQ doses (control, 1 and 5 µg/ml). Alkaline phosphatase activity and calcium measurements were performed to evaluate osteoblast differentiation and activity, respectively. Detailed microarray analysis was performed in 5 µg/ml HCQ‐treated cells and controls followed by qPCR validation. Additional cultures were performed using the cholesterol synthesis inhibitor simvastatin (SIM) to evaluate a potential mechanism of action. We showed that HCQ inhibits both MSC‐derived osteoblast differentiation and mineralization in vitro. Microarray analysis and additional PCR validation revealed a highly significant up‐regulation of the cholesterol biosynthesis, lysosomal and extracellular matrix pathways in the 5 µg/ml HCQ‐treated cells compared to controls. Besides, we demonstrated that 1 µM SIM also decreases MSC‐derived osteoblast differentiation and mineralization compared to controls. It appears that the positive effect of HCQ on BMD cannot be explained by a stimulating effect on the MSC‐derived osteoblast. The discrepancy between high BMD and decreased MSC‐derived osteoblast function due to HCQ treatment might be caused by systemic factors that stimulate bone formation and/or local factors that reduce bone resorption, which is lacking in cell cultures.     

Preparation and characterization of monoclonal antibodies to human adiponectin for its quantitative measurement in serum

Mouse Monoclonal antibodies against human adiponectin were produced by the routine method and the specificity of antibodies was verified. These monoclonal antibodies (MoAbs) interacted with the monomeric and trimeric forms of recombinant adiponectin according to the results of a Western blot analysis. Human blood serum was fractionated by gel filtration, and the protein of these fractions was stained using labeled MoAbs. It was established that a single high-molecular-weight form (HMW) of endogenous adiponectin was detected by this method. The use of competitive enzymelinked immunoassay on the basis of the obtained MoAbs allowed us to show that the sera of healthy male donors contains lower adiponectin concentrations than that of female donors (8.42 ± 1.59 μg/ml vs. 11.01 ± 2.58 μg/ml, p = 0.01). We also detected statistically significant lower adiponectin levels in the serum of patients with coronary artery disease for both men (6.01 ± 2.73 μg/ml vs. 8.42 ± 1.59 μg/ml, p = 0.015) and women (5.79 ± 2.98 μg/ml vs. 11.01 ± 2.58 μg/ml, p = 0.0003). Therefore, the developed methods for the analysis of the HMW form of adiponectin can be helpful in the diagnostics of the possible implications and assessment of unfavorable prognoses in patients with cardiovascular disorders.     

Human Melatonin and Cortisol Circadian Rhythms in Patients with Coronary Heart Disease

Cortisol and melatonin have well known circadian rhythms, coupled to the solar day. Melatonin has been shown to serve as an endogenuous “Zeitgeber” (time giver) and is secreted by the human pineal gland throughout the night but not during the day. Patients with coronary heart disease (CHD) have significant depressed nocturnal melatonin secretion compared to healthy individuals (Brugger et al., 1995). In addition to our previous study we measured serum concentrations of cortisol to evaluate whether the circadian rhythm of cortisol secretion is also different in patients with CHD. Blood was collected by venous puncture at 0200 and at 1400, serum separated and kept frozen at -20°C until analysis. Cortisol and melatonin were measured with a commercially available radioimmunoassay according to the instructions of the manufacturer. Nineteen patients with angiographically documented CHD (mean age 53 years) participated in this study. As control group served 12 adults without any signs of CHD. Melatonin serum concentrations (median; mean ± SD) at night were significantly depressed in patients with coronary heart disease (7.8; 8.6 ± 3.3 pg/ml) in comparison to the control group (38.0; 45.4 ± 24.1 pg/ml) p &lt; 0.01. Melatonin in the afternoon was not detectable in either of the groups. Cortisol values at night were significantly raised in patients with coronary heart disease (6.0; 7.2 ± 3.7 µg/dl) in comparison to the control group (2.7; 3.8 ± 2.9 µg/dl) p &lt; 0.05. Cortisol levels in the afternoon were also elevated in patients with CHD (8.9; 9.5 ± 3.8 µg/dl) but there was no significant difference compared to controls (6.8; 6.9 ± 4.5 µg/dl). The results of the present study indicate that patients with coronary heart disease have atypical secretory patterns of nocturnal cortisol and melatonin secretion.     

Adiponectin Improves Cardiomyocyte Contractile Function in db/db Diabetic Obese Mice

Low levels of adiponectin, a fat‐derived hormone, are found to be correlated with coronary heart disease, type 2 diabetes, obesity, and insulin resistance. Conversely, high adiponectin levels are predictive of reduced coronary risk in long‐term epidemiologic studies. However, the precise role of adiponectin in cardiomyocyte function is still not clear. This study was designed to examine the role of adiponectin in cardiac contractile function in the db/db model of diabetic obesity. Mechanical properties and intracellular Ca²⁺ transients were evaluated in cardiomyocytes from lean control and db/db mice with or without adiponectin (10 µg/ml) treatment. Expression and phosphorylation of IRS‐1, Akt, c‐Jun, and c‐Jun N terminal kinase (JNK) as well as markers of endoplasmic reticulum (ER) stress were evaluated using western blotting. Cardiomyocytes from db/db mice exhibited greater cross‐sectional area, depressed peak shortening (PS), and maximal velocity of shortening/re‐lengthening as well as prolonged duration of re‐lengthening. Consistently, myocytes from db/db mice displayed a reduced electrically stimulated rise in intracellular Ca²⁺ and prolonged intracellular Ca²⁺ decay, which were abrogated by adiponectin treatment. Ratios between phosphorylated c‐Jun and c‐Jun as well as phosphorylated IRS‐1 and IRS‐1 were increased in db/db mice, the effect of which was attenuated by adiponectin. Levels of the phosphorylated ER stress makers PERK (Thr980), IRE‐1, and eIF2α were significantly elevated in db/db mice compared with lean controls, although the effect was unaffected by adiponectin. Collectively, our data suggest that adiponectin improves cardiomyocyte dysfunction in db/db diabetic obese mice through a mechanism possibly related to c‐Jun and IRS‐1.     

Fluorimetric determination of proteins using 4‐chloro‐(2′‐hydroxylophenylazo)rhodanine–Ti(IV) complex as a spectral probe

A novel method for the determination of proteins was developed, based on the enhancement of fluorescence with 4‐chloro‐(2′‐hydroxylophenylazo)rhodanine–Ti(IV) [ClHARP–Ti(IV)] complex as a fluorescence probe. The excitation and emission wavelengths of the system were 335 nm and 376 nm, respectively. The presence of bis(2‐ethylhexyl)sulphosuccinate sodium salt (AOT) microemulsion greatly increased the sensitivity of the system. Under optimal conditions, four kinds of proteins, including bovine serum albumin (BSA), human serum albumin (HSA), egg albumin (Ova), and γ‐globin (γ‐G) were studied. The detection limits were 0.182 µg/mL for BSA, 0.0788 µg/mL for HSA, 0.216 µg/mL for Ova and 0.484 µg/mL for γ‐G. The linear ranges of the calibration were 0–12.0, 0–10.0, 0–18.0 and 0–18.0 µg/mL, respectively. The method possessed high sensitivity, good selectivity and was applied to the analysis of protein in milk powder and cornmeal with satisfactory results. Copyright © 2008 John Wiley & Sons, Ltd.     

The interplay of LncRNA ANRIL and miR‐181b on the inflammation‐relevant coronary artery disease through mediating NF‐κB signalling pathway

This study was designed to investigate whether ANRIL affected the aetiology of coronary artery disease (CAD) by acting on downstream miR‐181b and NF‐κB signalling. Altogether 327 CAD patients diagnosed by angiography were included, and mice models of CAD were established. Human coronary endothelial cells (HCAECs) and human umbilical vein endothelial cells (HUVECs) were also purchased. In addition, shRNA‐ANRIL, shRNA‐NC, pcDNA3.1‐ANRIL, miR‐181b mimic, miR‐181b inhibitor and miR‐NC were transfected into the cells. The lipopolysaccharides (LPS) and pyrrolidine dithiocarbamate (PDTC) were also added to activate or deactivate NF‐κB signalling. Both highly expressed ANRIL and lowly expressed miR‐181b were associated with CAD population aged over 60 years old, with smoking history, with hypertension and hyperlipidemia, with CHOL H 4.34 mmol/L, TG ≥ 1.93 mmol/L and Hcy ≥ 16.8 μmol/L (all P < 0.05). Besides, IL‐6, IL‐8, NF‐κB, TNF‐α, iNOS, ICAM‐1, VCAM‐1 and COX‐2 expressions observed within AD mice models were all beyond those within NC and sham‐operated groups (P < 0.05). Also VEGF and HSP 70 were highly expressed within AD mice models than within NC and sham‐operated mice (P < 0.05). Transfection of either pcDNA‐ANRIL or miR‐181b inhibitor could significantly fortify HCAECs’ viability and put on their survival rate. At the meantime, the inflammatory factors and vascular‐protective parameters were released to a greater level (P < 0.05). Finally, highly expressed ANRIL also notably bring down miR‐181b expression and raise p50/p65 expressions within HCAECs (P < 0.05). The joint role of ANRIL, miR‐181b and NF‐κB signalling could aid in further treating and diagnosing CAD.     

Association of plasma exosomes with severity of organ failure and mortality in patients with sepsis

Current sepsis biomarkers may be helpful in determining organ failure and evaluating patient clinical course; however, direct molecular biomarkers to predict subsequent organ failure have not yet been discovered. Exosomes, a small population of extracellular vesicles, play an important role in the inflammatory response, coagulation process and cardiac dysfunction in sepsis. Nonetheless, the association of plasma exosome with severity and mortality of sepsis is not well known. Therefore, the overall levels of plasma exosome in sepsis patients were assessed and whether exosome levels were associated with organ failure and mortality was evaluated in the present study. Plasma level of exosomes was measured by ELISA. Among 220 patients with sepsis, 145 (66%) patients were diagnosed with septic shock. A trend of increased exosome levels in control, sepsis and septic shock groups was observed (204 µg/mL vs 525 µg/mL vs 802 µg/mL, P < 0.001). A positive linear relationship was observed between overall exosome levels and Sequential Organ Failure Assessment (SOFA) score in the study cohorts (r value = 0.47). When patients were divided into two groups according to best cut‐off level, a statistical difference in 28‐ and 90‐day mortality between patients with high and low plasma exosomes was observed. Elevated levels of plasma exosomes were associated with severity of organ failure and predictive of mortality in critically ill patients with sepsis.     

Chalcone analogues: Synthesis,activity against Meloidogyne incognita,and in silico interaction with cytochrome P450

To contribute the development of new products to control plant‐parasitic nematodes, 12 chalcone analogues were synthesized and screened for activity against Meloidogyne incognita. Three caused mortality greater than negative controls in second‐stage juvenile M. incognita, with values varying from 19.9% to 100%. The most active chalcone analogue was (1E,4E)‐1,5‐di(4‐nitrophenyl)‐2‐butylpenta‐1,4‐dien‐3‐one (compound 6 ), which had an LC₅₀ value of 41 µg/ml. Under the same conditions, the commercial nematicide Carbofuran^® (2,2‐dimethyl‐2,3‐dihydro‐1‐benzofuran‐7‐yl methylcarbamate) presented an LC₅₀ equal to 101 µg/ml. When this chalcone analogue was applied to tomato plants infested with M. incognita, reductions in the numbers of galls and eggs of 51% and 68% were observed, respectively. According to in silico studies, the enzyme target of compound 6 in M. incongita is cytochrome P450, which is important for the oxidation of several substances in the nematode. Therefore, compound 6 is potentially useful for the development of new products to control M. incongita.     

Elevated Insulin Sensitivity in Low‐protein Offspring Rats Is Prevented by a High‐fat Diet and Is Associated With Visceral Fat

This study tests the hypothesis that a high‐fat postnatal diet increases fat mass and reduces improved insulin sensitivity (IS) found in the low‐protein model of maternal undernutrition. Offspring from Wistar dams fed either a 20% (control (CON)) or 8% (low protein (LP)) protein diet during gestation and lactation were randomly assigned to a control (con) or cafeteria (caf) diet at weaning (21 days) until 3 months of age at which point IS was measured (hyperinsulinemic–euglycemic clamp). Fat mass, growth, energy intake (EI) and expenditure (EE), fuel utilization, insulin secretion, and leptin and adiponectin levels were measured to identify a possible role in any changes in IS. IS was increased in LP‐con in comparison to CON‐con animals. Cafeteria feeding prevented this increase in LP animals but had no effect in CON animals (insulin‐stimulated glucose infusion rates (GIRs; mg/min/kg); CON‐con: 13.9 ± 1.0, CON caf: 12.1 ± 2.1, LP‐con: 25.4 ± 2.0, LP‐caf: 13.7 ± 3.7, P < 0.05). CON‐caf animals had similar percent epididymal white adipose tissue (%EWAT; CON‐con: 1.71 ± 0.09 vs. CON‐caf: 1.66 ± 0.08) and adiponectin (µg/ml: CON‐con: 4.61 ± 0.34 vs. CON‐caf: 3.67 ± 0.18) except hyperinsulinemia and relative hyperleptinemia in comparison to CON‐con. Differently, LP‐caf animals had increased %EWAT (LP‐con: 1.11 ± 0.06 vs. LP‐caf: 1.44 ± 0.08, P < 0.05) and adiponectin (µg/ml: LP‐con: 5.38 ± 0.39 vs. LP‐caf: 3.75 ± 0.35, P < 0.05) but did not show cafeteria‐induced hyperinsulinemia or relative hyperleptinemia. An increased propensity to store visceral fat in LP animals may prevent the elevated IS in LP offspring.     

Eicosapentaenoic Acid and Rosiglitazone Increase Adiponectin in an Additive and PPARγ‐Dependent Manner in Human Adipocytes

Adiponectin, an anti‐inflammatory and insulin‐sensitizing protein secreted from adipose tissue, may be modulated by dietary fatty acids, although the mechanism is not fully known. Our objective was to investigate the effect of long‐chain n‐3 polyunsaturated fatty acids (PUFAs) on adiponectin in cultured human adipocytes, and to elucidate the role of peroxisome proliferator‐activated receptor‐γ (PPARγ) in this regulation. Isolated human adipocytes were cultured for 48 h with 100 µmol/l eicosapentaenoic acid (C20:5n‐3, EPA), docosahexaenoic acid (C22:6n‐3, DHA), palmitic acid (C16:0), 100 µmol/l EPA plus 100 µmol/l DHA, or bovine serum albumin (control). Additionally, adipocytes were treated for 48 h with a PPARγ antagonist (BADGE) or agonist (rosiglitazone) in isolation or in conjunction with either EPA or DHA. At 48 h, EPA and DHA increased (P < 0.05) adiponectin secretion by 88 and 47%, respectively, while EPA, but not DHA, also increased (136%, P < 0.001) cellular adiponectin protein. Interestingly, PPARγ antagonism completely abolished the DHA‐mediated increase in secreted adiponectin, but only partially attenuated the EPA‐mediated response. Thus, EPA's effects on adiponectin do not appear to be entirely PPARγ mediated. Rosiglitazone increased (P < 0.001) the secreted and cellular adiponectin protein (90 and 582%, respectively). Finally, the effects of EPA and rosiglitazone on adiponectin secretion were additive (+230% at 48 h combined, compared to 121 and 124% by EPA or rosiglitazone alone, respectively). Overall, our findings emphasize the therapeutic importance of long‐chain n‐3 PUFA alone, or in combination with a PPARγ agonist, as a stimulator of adiponectin, a key adipokine involved in obesity and related diseases.     

Effect of Fenofibrate on Adiponectin and Inflammatory Biomarkers in Metabolic Syndrome Patients

Adiponectin is an adipose‐secreted hormone with anti‐inflammatory properties mediated by inhibition of nuclear factor‐κB (NF‐κB) signaling. This study investigates whether fenofibrate alters adiponectin levels in patients with hypertriglyceridemia and the metabolic syndrome, and examines the association of adiponectin with circulating inflammatory markers and whole blood cytokine production. The effects of fenofibrate (160 mg/day) on adiponectin and other inflammatory markers were investigated in a 12‐week randomized, placebo‐controlled trial in 55 patients with hypertriglyceridemia (plasma triglycerides ≥1.7 mmol/l and <6.8 mmol/l), central obesity and other characteristics of the metabolic syndrome who were not receiving lipid‐altering therapies. In the fenofibrate group, adiponectin levels increased from 4.10 to 4.50 µg/ml (+7.7%); in the placebo group, adiponectin levels increased by 1.8%; (P = 0.0005). In multivariate models including age, gender, and waist circumference, there were inverse correlations between changes in adiponectin and vascular cell adhesion molecule‐1 (VCAM‐1) (r = −0.54, P < 0.0001) and intercellular adhesion molecule‐1 (ICAM‐1) (r = −0.57, P < 0.0001), and C‐reactive protein (CRP) (r = −0.40, P = 0.0041); lipopolysaccharide (LPS)‐stimulated production of tumor necrosis factor‐α (TNF‐α) (r = −0.30, P = 0.035), interleukin (IL)‐1β (r = −0.44, P = 0.0016), monocyte chemotactic protein‐1 (MCP‐1) (r = −0.46, P = 0.001), and macrophage inflammatory protein‐1α (MIP‐1α) (r = −0.45, P = 0.0012). Fenofibrate (160 mg/day) raised adiponectin levels in patients with hypertriglyceridemia and the metabolic syndrome. Changes in adiponectin were significantly and inversely associated with changes in multiple inflammatory markers. These data suggest that adiponectin may contribute to the anti‐inflammatory effects of fenofibrate.     

Purification and partial characterization of a novel lectin from Dioclea lasiocarpa Mart seeds with vasodilator effects

A lectin from seeds of Dioclea lasiocarpa (DLL) was purified in a single step by affinity chromatography in a Sephadex G‐50 column. DLL haemagglutinated rabbit erythrocytes showing stability even after 1 h of exposure to a different pH values (optimal between pH 6.0 and 8.0) but was inhibited after incubation with d ‐mannose and d ‐glucose. The pure protein possessed a molecular weight of 25 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis and 25,410Da by mass spectrometry. The results analyzed by the software SELCON 3 indicate that β‐sheet secondary structures are predominant in DLL (approximately 40.2% antiparallel β‐sheet, 4.6% parallel β‐sheet, 7.2% α‐helices, 17.3% turns, and 28.7% unordered structures). Mechanical activity of isolated aorta from rat measured by cumulative concentration curves of DLL, performed at the contraction plateau induced by phenylephrine in either endothelium‐intact or denuded aorta. DLL (IC₅₀ = 34.12 ± 3.46 µg/ml) relaxed precontracted endothelized aortic rings by 34.61 ± 9.06%, 55.19 ± 11.9%, and 81.33 ± 14.35%, respectively, at 10 µg/ml (initial concentration), 30 µg/ml, and 100 µg/ml (maximum effect). All effects occurred via interaction with lectin domains and participation of nitric oxide. Copyright © 2012 John Wiley & Sons, Ltd.     

Decrease in Serum Adiponectin Level Due to Obesity and Visceral Fat Accumulation in Children

Objective: To determine whether serum adiponectin is decreased in obesity and is restored toward normal level after treatment in children. Research Methods and Procedures: Subjects were 53 Japanese obese children, 33 boys and 20 girls (6 to 14 years old), and 30 age‐matched nonobese controls for measuring adiponectin (16 boys and 14 girls). Blood was drawn after an overnight fast, and the obese children were subjected to anthropometric measurements including waist and hip circumferences and skinfold thicknesses. Paired samples were obtained from 21 obese children who underwent psychoeducational therapy. Visceral adipose tissue area was measured by computed tomography. Adiponectin was assayed by an enzyme‐linked immunosorbent assay. Results: The serum levels of alanine aminotransferase, uric acid, triglyceride, total cholesterol, low‐density lipoprotein‐cholesterol, total cholesterol/high‐density lipoprotein‐cholesterol, apo B, apo B/apo A₁, and insulin in obese children were higher than the reference values. Serum adiponectin level was lower in the obese children than in the controls (6.4 ± 0.6 vs. 10.2 ± 0.8 mg/L, means ± SEM, p < 0.001). In 21 obese children whose percent overweight declined during therapy, the adiponectin level increased (p = 0.002). The adiponectin level was correlated inversely with visceral adipose tissue area in obese children (r = ?0.531, p < 0.001). The inverse correlations of adiponectin with alanine aminotransferase, uric acid, and insulin were significant after being adjusted for percentage overweight, percentage body fat, or sex. Discussion: Serum adiponectin level is decreased in obese children depending on the accumulation of visceral fat and is restored toward normal level by slimming.     

Detection and molecular characterization of carbendazim-resistant Colletotrichum truncatum Isolates causing anthracnose of soybean in Thailand

A loss of fungicide efficacy, particularly for carbendazim, was noted in soybean fields in Thailand and was considered to be due to the development of Colletotrichum truncatum resistance. The carbendazim sensitivity of C. truncatum populations isolated from various soybean fields in Thailand was thus evaluated with in vitro sensitivity assays and molecular characterization of mutations in the sequences of the ß₂-tubulin (TUB2) gene that confer carbendazim resistance in the pathogen. Among 52 isolates, 46 isolates were classified as highly resistant (HR) to carbendazim (EC₅₀ > 1,000 µg/ml). All HR isolates grew on PDA amended with carbendazim at 1,000 µg/ml. Six isolates were classified as carbendazim sensitive (S) (EC₅₀ < 1 µg/ml). Mycelial growth on PDA amended with 1 µg/ml carbendazim was inhibited by over 50% compared with growth on PDA alone. When a partial TUB2 gene from the isolates was amplified and analysed using predicted amino acid sequences, an alteration from glutamic acid to alanine at codon 198 (E198A) was found in 45 HR isolates for which the EC₅₀ was higher than 2000 µg/ml. This mutation resulted from a nucleotide substitution from adenine to cytosine (GA G → GC G). The other HR isolate, CtPhS_1, with EC₅₀ of 1,127 µg/ml, had an alteration at codon 200 (F200Y) (TT C → TA C).     

Piper crocatum Ruiz & Pav. ameliorates wound healing through p53, E-cadherin and SOD1 pathways on wounded hyperglycemia fibroblasts

IntroductionPiper crocatum Ruiz & Pav (P. crocatum) has been reported to accelerate the diabetic wound healing process empirically. Some studies showed the benefits of P. crocatum in treating various diseases but its mechanisms in diabetic wound healing have never been reported. In the present study we investigated the diabetic wound healing activity of the active fraction of P. crocatum on wounded hyperglycemia fibroblasts (wHFs).MethodsBioassay-guided fractionation was performed to get the most active fraction. The selected active fraction was applied to wHFs within 72 h incubation. Mimicking a diabetic condition was done using basal glucose media containing an additional 17 mMol/L D-glucose. A wound was simulated via the scratch assay. The collagen deposition was measured using Picro-Sirius Red and wound closure was measured using scratch wound assay. Underlying mechanisms through p53, αSMA, SOD1 and E-cadherin were measured using western blotting.ResultsWe reported that F_IV is the most active fraction of P. crocatum. We confirmed that F_IV\(7.81 µg/ml, 15.62 µg/ml, 31.25 µg/ml, 62.5 µg/ml, and 125 µg/ml) induced the collagen deposition and wound closure of wHFs. Furthermore, F_IV treatment (7.81 µg/ml, 15.62 µg/ml, 31.25 µg/ml) down-regulated the protein expression level of p53 and up-regulated the protein expression levels of αSMA, E-cadherin, and SOD1.Discussion/conclusionsOur findings suggest that ameliorating collagen deposition and wound closure through protein regulation of p53, αSMA, E-cadherin, and SOD1 are some of the mechanisms by which F_IV of P. crocatum is involved in diabetic wound healing therapy.     

Antifungal Susceptibility Profile of Candida Albicans Isolated from Vulvovaginal Candidiasis in Xinjiang Province of China

We investigated the antifungal susceptibility profiles of 207 independent Candida albicans strains isolated from patients with vulvovaginal candidiasis (VVC) in Xinjiang Province of China. Using CLSI M27-A3 and M27-S4 guidelines, anidulafungin and micafungin were the most active drugs against C. albicans showing an MIC₅₀/MIC₉₀ corresponding to 0.016/0.0313 µg/mL, followed by caspofungin (0.25/0.25 µg/mL), posaconazole (0.125/0.5 µg/mL), ravuconazole (0.063/1 µg/mL), itraconazole (0.125/1 µg/mL), amphotericine B (0.5/1 µg/mL), isavuconazole (0.063/2 µg/mL), 5-flucytosine (1/2 µg/mL), voriconazole (0.125/4 µg/mL), and fluconazole (0.5/4 µg/mL). 96.1% (199)–100.0% (207) isolates were sensitive to the three echinocandins tested, amphotericine B and 5-flucytosine. The in vitro activity of triazoles against all isolates tested was variable; itraconazole and voriconazole had reduced the activity to almost half of the isolates (55.1% (114) and 51.2% (106) susceptible, respectively). Fluconazole was active against 76.3% (158) isolates tested. The new triazoles ravuconazole, isavuconazole and posaconazole showed good in vitro potency against 89.9% (186)–95.2% (197) of isolates with the geometric mean MIC (µg/mL) of 0.10, 0.12 and 0.14 µg/mL, respectively. In conclusion, our study indicates that for effective management of systemic candidiasis in Xinjiang Province of China, it is important to determine the susceptibility profiles of isolated C. albicans from patients with VVC.