Molecular identification of non-Cryptococcus yeasts associated with pigeon droppings in Shiraz, Southern Iran.

BACKGROUND: Birds are considered as a reservoir for pathogenic and non-pathogenic fungi. Pigeon droppings have the potential for spreading these fungi to the environment. Cryptococcus species are important fungi associated with pigeon droppings. In this regard, there are many types of yeast associated with guano that is important for human and animal health. AIMS: The main objective of this study is the identification of non-Cryptococcus yeasts isolated from pigeon dropping in Shiraz, Southern Iran. METHODS: A total of 100 unknown yeasts, which were previously screened and identified as non-Cryptococcus from pigeon guano through the conventional methods, were used in this study. Identification of the isolates was performed based on conventional methods and DNA sequence analysis of internal transcribed spacer (ITS) rDNA gene region. The sequence results were deposited in NCBI database using the Basic Local Alignment Search Tool (BLAST). RESULTS: A total of 16 species belonging to 7 genera were identified as Candida spp. 51% (8 species), Rhodotorula sp. 24%, Trichosporon spp. 21% (3 species), Rhodosporidium 2%, Saccharomyces 1%, Rhizoctonia 1%, and Meyerozyma 1%. The predominant isolates were Rhodotorula rubra (24%), Candida famata (20%), and Trichosporon asahii (13%). The other species were Rhodosporidium kratochvilovae 2 (2%), Saccharomyces cerevisiae 1 (1%), Rhizoctonia solani 1 (1%), and Meyerozyma caribbica 1 (1%). CONCLUSION: Pigeon excreta examined in this study were associated with several kinds of opportunistic yeasts which could cause diseases in prone human and animals.

Regular consumption of both vitamin D- and calcium- and vitamin D-fortified yogurt drink is equally accompanied by lowered blood lipoprotein (a) and elevated apoprotein A1 in subjects with type 2 diabetes: a randomized clinical trial.

OBJECTIVE: Cardiovascular disease (CVD) is the major morbidity and cause of death in diabetic subjects. Observational studies have shown the association of low vitamin D status with poor glycemic control, atherogenic lipid profile, and CVD. However, the possible link between circulating 25-hydroxycholecalciferol and apoproteins (Apo A1 and B) and the atherogenic lipoprotein (a) [Lp(a)] has not been documented to date. METHODS: Ninety subjects with type 2 diabetes (T2D) aged 30-60 years from both sexes were randomly allocated to one of the 3 groups to receive 2 bottles a day of either (1) plain doogh (PD; containing 150 mg calcium and no detectable vitamin D/250 mL); (2) vitamin D-fortified doogh (DD; containing 150 mg calcium and 500 IU vitamin D/250 mL); or (3) calcium- and vitamin D-fortified doogh (CDD; containing 250 mg calcium and 500 IU vitamin D/250 mL) for 12 weeks. Anthropometric, dietary, and laboratory assessments, including Apo A1, Apo B, and Lp(a), were done. RESULTS: Improvement of vitamin D status in DD and CDD groups, compared to PD, resulted in a significant increase in Apo A1 (mean changes 0.22 ± 0.38, 0.20 ± 0.27 and 0.01 ± 0.35 g/L, respectively, p = 0.047) and a significant decrease in serum Lp(a) (mean changes -0.08 ± 0.30, -0.08 ± 0.31, and 0.14 ± 0.25 µmol/L, respectively, p = 0.011). There was no significant difference between DD and CDD groups. Serum Apo B did not change significantly in any of the groups. CONCLUSIONS: Significant amelioration of serum Apo A1 and Lp(a) following improvement of vitamin D status in T2D subjects may have preventive implications against long-term diabetic complications, notably CVD. This trial was registered at ClinicalTrials.gov as NTC01229891.

Vitamin D Receptor Fok-I polymorphism modulates diabetic host response to vitamin D intake: need for a nutrigenetic approach.

OBJECTIVE: Interpopulation as well as interindividual variations in response to vitamin D intake commonly observed in subjects with type 2 diabetes may be related to genetic makeup. One of the candidate genes potentially responsible for this diversity is vitamin D receptor (VDR). This study aimed to investigate the interactive effect of VDR Fok-I polymorphism and vitamin D intake on diverse aspects of diabetic host response. RESEARCH DESIGN AND METHODS: Glycemic status, lipid profiles, inflammatory biomarkers, and VDR Fok-I genotypes were determined in diabetic subjects (n = 140) who participated in a randomized controlled trial. Participants consumed two 250-mL bottles per day of yogurt drink (doogh) fortified with 500 IU vitamin D/250 mL for 12 weeks. RESULTS: Mean serum 25(OH)D increased by ~30 nmol/L (P < 0.001). The time × intervention effect was significant for 25(OH)D (P = 0.030), HDL (P = 0.011), high-sensitivity C-reactive protein (hsCRP) (P < 0.001), interleukin (IL)-4 (P = 0.008), and IL-6 (P = 0.017) among the genotypic groups. The alleles were defined as ''F'' or ''f'' depending on the absence or presence of the restriction site, respectively. The least increment in 25(OH)D was in ff (23.0 ± 3.8 nmol/L) compared with Ff (31.2 ± 3.4 nmol/L) and FF (35.6 ± 2.7 nmol/L) (P for trend = 0.009), but only the difference between ff and FF was significant (P = 0.023). FF group had the largest decrement of both hsCRP and IL-6 compared with Ff (P < 0.001 and P = 0.038) and ff (P = 0.010 and P = 0.048), respectively. CONCLUSIONS: We concluded that those of VDR ff genotype may be regarded as "low responders" to vitamin D intake in terms of response of circulating 25(OH)D and certain inflammatory biomarkers. A nutrigenetic approach may, therefore, be needed to protect diabetic patients from vitamin D deficiency.

Vitamin D status and the predictors of circulating T helper 1-type immunoglobulin levels in Iranian subjects with type 1 diabetes and their siblings: a case-control study.

BACKGROUND: Vitamin D status has been linked to both T helper (Th)1/Th2 balance and susceptibility to type 1 diabetes (T1D). The present study aimed to evaluate vitamin D status and its relation to Th1/Th2 balance in subjects with T1D, their siblings and unrelated healthy controls during autumn and winter 2008-2009. METHODS: A case-control study was conducted on subjects with T1D (n(1) = 60) and two control groups comprising nondiabetic siblings (n(2) = 60) and unrelated healthy controls (n(3) = 60). Assessments of dietary intake, anthropometry, intact parathyroid hormone (iPTH) and 25(OH)D were performed. Serum levels of immunoglobulin (Ig)G(2) and IgE, as well as the IgG2/IgE ratio, were used to evaluate Th1/Th2 balance. Vitamin D status was defined based on circulating 25(OH)D as deficiency: <27.5 nm; insufficiency 27.5 ≤ 25(OH)D <50 nm; and sufficiency ≥50 nm. RESULTS: Vitamin D status did not differ significantly among the groups. Similarly, no significant difference in 25(OH)D, iPTH, IgG(2), IgE and IgG(2)/IgE was found. In multiple regression analysis of pooled data, PTH and body mass index were the predictors of IgG(2)/IgE. In the diabetic group, both PTH and age and, in siblings, PTH only, were the predictors of IgG(2)/IgE ratio. CONCLUSIONS: These data suggest PTH as the major predictor of immune deviance towards the Th1 response in both type 1 diabetic subjects and their siblings. Considering that the active form of vitamin D suppresses PTH production, it is hypothesised that vitamin D replenishment of just those who are genetically prone to the disease (i.e. siblings) may be regarded as a preventive measure against T1D.

Improvement of vitamin D status resulted in amelioration of biomarkers of systemic inflammation in the subjects with type 2 diabetes.

BACKGROUND: Both vitamin D deficiency and inflammation have been linked to cardiovascular disease, the major cause of death in diabetes. In this study, the effects of daily intake of vitamin D-fortified yoghourt drink (doogh) on systemic inflammatory biomarkers in subjects with type 2 diabetes (T2D) were investigated. SUBJECTS AND METHODS: In this 12-week randomized controlled trial, T2D subjects received either plain doogh (PD; containing 170 mg calcium and no detectable vitamin D/250 mL, n(1) = 50) or vitamin D3-fortified doogh (FD; containing 170 mg calcium and 500 IU/250 mL, n(2) = 50) twice a day. Glycemic status, body fat mass and systemic inflammatory biomarkers including serum highly sensitive C-reactive protein (hsCRP), serum amyloid A (SAA), interleukin(IL)-2, IL-6, IL-10 and tumour necrosis factor (TNF)-α were evaluated at the beginning and after the intervention. Data were expressed as either mean ± SD or median (interquartile range) whenever they had either normal or non-normal distribution, respectively. RESULTS: In the patients receiving the vitamin D fortified drink, compared with those receiving the unfortified drink, a significant increase in serum 25(OH)D was accompanied by significant changes in TNF-α (-57.9 (-264.6) versus +106.3 (683.2), p = 0.044), IL-6 (-6.3 (-69.2), p = 0.002), hsCRP (-0.39 (-1.50) versus +0.8 (1.52), p < 0.001), SAA (-14.2 ± 44.5 versus +5.6 ± 37.5 mg/L, p = 0.022) and IL-10 (+38.7 ± 157.0 versus -51.9 ± 165.2 ng/L, p = 0.013). The between-group differences of hsCRP, SAA and IL-6 changes remained significant even after controlling for changes quantitative insulin check index (p < 0.001, p < 0.001 and p = 0.009, respectively). CONCLUSIONS: Improvement of vitamin D status of T2D subjects resulted in amelioration of the systemic inflammatory markers. This may have preventive implications against cardiovascular disease and other diabetic complications.

High prevalence of vitamin D deficiency in school-age children in Tehran, 2008: a red alert.

OBJECTIVE: To assess the vitamin D status of 9-12-year-old primary-school children in Tehran during autumn and winter 2007-2008. DESIGN: A descriptive cross-sectional study. SETTING: Primary schools of Tehran city, Iran. SUBJECTS: A total of 1111 children aged 9-12 years (573 boys and 538 girls) from sixty primary schools were enrolled in the study. Weight, height, BMI and serum levels of Ca, P, Mg, 25-hydroxyvitamin D (25(OH)D), intact parathyroid hormone (iPTH), osteocalcin and bone-specific alkaline phosphatase of all the participants were assessed. Dietary Ca intake was also evaluated using a quantitative FFQ for a subsample of the study population (n 503). Vitamin D sufficiency was defined on the basis of serum levels of 25(OH)D as either ≥37 nmol/l (criterion 1) or ≥50 nmol/l (criterion 2). RESULTS: Daily intake of Ca did not differ significantly between boys and girls (929·6 (sd 436·7) mg and 909·5 (sd 465·5) mg, respectively). However, on the basis of the first criterion, approximately 86 % of the children had vitamin D deficiency, with 38·3 % being severely deficient (25(OH)D < 12·5 nmol/l). According to the second criterion, prevalence of vitamin D deficiency rose to 91·7 %. Prevalence of vitamin D deficiency was higher in girls than in boys by either criterion. Serum levels of 25(OH)D inversely correlated with iPTH (r = -0·154, P < 0·001) and BMI (r = -0·092, P = 0·002) but directly correlated with duration of sun exposure (r = 0·115, P < 0·001). CONCLUSIONS: The high prevalence of vitamin D deficiency among schoolchildren (especially among girls) warrants immediate interventions for proper nutritional support.

Regular consumption of vitamin D-fortified yogurt drink (Doogh) improved endothelial biomarkers in subjects with type 2 diabetes: a randomized double-blind clinical trial.

BACKGROUND: Endothelial dysfunction has been proposed as the underlying cause of diabetic angiopathy that eventually leads to cardiovascular disease, the major cause of death in diabetes. We recently demonstrated the ameliorating effect of regular vitamin D intake on the glycemic status of patients with type 2 diabetes (T2D). In this study, the effects of improvement of vitamin D status on glycemic status, lipid profile and endothelial biomarkers in T2D subjects were investigated. METHODS: Subjects with T2D were randomly allocated to one of the two groups to receive either plain yogurt drink (PYD; containing 170 mg calcium and no vitamin D/250 mL, n1 = 50) or vitamin D3-fortified yogurt drink (FYD; containing 170 mg calcium and 500 IU/250 mL, n2 = 50) twice a day for 12 weeks. Anthropometric measures, glycemic status, lipid profile, body fat mass (FM) and endothelial biomarkers including serum endothelin-1, E-selectin and matrix metalloproteinase (MMP)-9 were evaluated at the beginning and after the 12-week intervention period. RESULTS: The intervention resulted in a significant improvement in fasting glucose, the Quantitative Insulin Check Index (QUICKI), glycated hemoglobin (HbA1c), triacylglycerols, high-density lipoprotein cholesterol (HDL-C), endothelin-1, E-selectin and MMP-9 in FYD compared to PYD (P < 0.05, for all). Interestingly, difference in changes of endothelin-1, E-selectin and MMP-9 concentrations in FYD compared to PYD (-0.35 ± 0.63 versus -0.03 ± 0.55, P = 0.028; -3.8 ± 7.3 versus 0.95 ± 8.3, P = 0.003 and -2.3 ± 3.7 versus 0.44 ± 7.1 ng/mL, respectively, P < 0.05 for all), even after controlling for changes of QUICKI, FM and waist circumference, remained significant for endothelin-1 and MMP-9 (P = 0.009 and P = 0.005, respectively) but disappeared for E-selectin (P = 0.092). On the contrary, after controlling for serum 25(OH)D, the differences disappeared for endothelin-1(P = 0.066) and MMP-9 (P = 0.277) but still remained significant for E-selectin (P = 0.011). CONCLUSIONS: Ameliorated vitamin D status was accompanied by improved glycemic status, lipid profile and endothelial biomarkers in T2D subjects. Our findings suggest both direct and indirect ameliorating effects of vitamin D on the endothelial biomarkers. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01236846.

Is vitamin D status a determining factor for metabolic syndrome? A case-control study.

This study was undertaken to assess vitamin D status in nonmenopausal women with metabolic syndrome (MeS) and to evaluate its possible role in inflammation and other components of MeS. A case-control study was conducted during late fall and winter 2009-10. A total of 375 women with waist circumference (WC) ≥88 cm were examined to find 100 who met MeS criteria according to the National Cholesterol Education Program (NCEP)/Adult Treatment Panel (ATP) III criteria (NCEP/ATP III). Of those without MeS, 100 age- and residence area-matched women were selected as a control group. Anthropometric and laboratory evaluations were performed. Waist-to-hip ratio (WHR), body mass index (BMI), homeostatic model of insulin resistance (HOMA-IR) and body fat mass (FM) were also evaluated. Women with MeS had significantly higher BMI, waist circumference (WC) and FM but lower serum osteocalcin than controls. There was no significant difference in serum 25 hydroxyvitamin D (25[OH]D), intact parathyroid hormone (iPTH) or vitamin D status between the two groups. Serum highly sensitive C-reactive protein (hsCRP) concentration was significantly higher in the MeS group, compared to the controls (3.4 ± 3.3 vs 2.0 ± 1.9 mg/L, P < 0.001). The difference remained significant even after controlling for BMI (P = 0.011), WC (P = 0.014) and FM (P = 0.005). When comparison was made only in those subjects with insulin resistance (HOMA-IR > 2.4), hsCRP was still higher in the MeS group (n = 79) than in the control group (n = 61) (P < 0.001). When data were categorized according to vitamin D status, in the MeS group significantly higher plasma glucose concentrations were observed in subjects with vitamin D deficiency compared to those with insufficiency or sufficiency (104.0 ± 11.7, 83.0 ± 11.3 and 83.2 ± 9.9 mg/dL, respectively, P < 0.001). Interestingly, their WC or WHR did not show any significant difference. In stepwise regression analysis, 25(OH)D was the main predictor of both hsCRP and plasma glucose. Vitamin D status may, at least in part, be a determining factor of systemic inflammation and the related metabolic derangements of MeS.

Daily consumption of vitamin D- or vitamin D + calcium-fortified yogurt drink improved glycemic control in patients with type 2 diabetes: a randomized clinical trial.

BACKGROUND: Low serum concentrations of 25-hydroxyvitamin D [25(OH)D] have been associated with impaired glucose tolerance and diabetes. OBJECTIVE: This study aimed to compare the effects of daily intake of vitamin D- or vitamin D(3) + calcium-fortified yogurt drink on glycemic status in subjects with type 2 diabetes (T2D). DESIGN: Ninety diabetic subjects were randomly allocated to 3 groups to consume plain yogurt drink (PY; containing no vitamin D and 150 mg Ca/250 mL), vitamin D-fortified yogurt drink (DY; containing 500 IU vitamin D(3) and 150 mg Ca/250 mL), or vitamin D + calcium-fortified yogurt drink (DCY; containing 500 IU vitamin D(3) and 250 mg Ca/250 mL) twice per day for 12 wk. Fasting serum glucose (FSG), glycated hemoglobin (Hb A(1c)), homeostasis model assessment of insulin resistance (HOMA-IR), serum lipid profile, and percentage fat mass (FM) were assessed before (baseline) and after the intervention. RESULTS: In both the DY and DCY groups, mean serum 25(OH)D(3) improved (+32.8 ± 28.4 and +28.8 ± 16.1 nmol/L, respectively; P < 0.001 for both), but FSG [-12.9 ± 33.7 mg/dL (P = 0.015) and -9.6 ± 46.9 mg/dL (P = 0.035)], Hb A(1c) [-0.4 ± 1.2% (P < 0.001) and -0.4 ± 1.9% (P < 0.001)], HOMA-IR [-0.6 ± 1.4 (P = 0.001) and -0.6 ± 3.2 (P < 0.001)], waist circumference (-3.6 ± 2.7 and -2.9 ± 3.3; P < 0.001 for both), and body mass index [in kg/m(2); -0.9 ± 0.6 (P < 0.001) and -0.4 ± 0.7 (P = 0.005)] decreased significantly more than in the PY group. An inverse correlation was observed between changes in serum 25(OH)D(3) and FSG (r = -0.208, P = 0.049), FM (r = -0.219, P = 0.038), and HOMA-IR (r = -0.219, P = 0.005). CONCLUSION: Daily intake of a vitamin D-fortified yogurt drink, either with or without added calcium, improved glycemic status in T2D patients. This trial was registered at clinicaltrials.gov as NCT01229891.

Regular daily intake of black tea improves oxidative stress biomarkers and decreases serum C-reactive protein levels in type 2 diabetic patients.

BACKGROUND: This study was undertaken to evaluate the possible effects of different daily doses of black tea intake on certain oxidative stress, inflammatory and metabolic biomarkers in patients with type 2 diabetes mellitus (T2DM). METHODS: Forty-six patients with known T2DM were randomly assigned either to the test (n = 23, 57.0 +/- 7.9 years) or the control (n = 23, 55.4 +/- 8.3 years) group. Following a one-week 'run-in' period, the test group received 150, 300, 450 and 600 ml of black tea extract (BTE) during the weeks 1, 2, 3 and 4, respectively. The control group received 150 ml BTE a day throughout the intervention period. Dietary, anthropometric and biochemical assessments were performed at the end of each week. FINDINGS: Serum total antioxidant capacity was enhanced similarly in both test and control groups. However, daily intake of 2 cups of BTE by the test group showed a suppressing effect on serum malondialdehyde. Serum C-reactive protein significantly decreased and glutathione levels increased following the intake of 4 cups (600 ml) of BTE a day. CONCLUSION: Regular intake of BTE had anti-oxidative and anti-inflammatory effects in patients with T2DM. These findings may, to some extent, explain the mechanisms underlying the protective effects of drinking tea against cardiovascular disease.

Familial IgA nephropathy in southeastern Kentucky.

BACKGROUND: Two decades ago, pedigrees of patients with IgA nephropathy (IgAN) from Pike County, KY, USA, provided evidence for a role of genetic factors in the pathogenesis of this disorder. Subsequently additional pedigrees were described for several communities from northern Italy. Recently, we found another cluster of patients in the Clay County, KY area, about 100 miles southwest of Pike County. AIM: The purpose of this study was to evaluate and expand the pedigrees of patients with IgAN from Clay County, KY to provide additional insight into the mechanisms of inheritance of IgAN and assess the possible influence of a founder effect on the prevalence of IgAN in the region. METHOD: Since 1980, most patients with IgAN and their relatives in eastern KY have provided personal genealogic data. These data were used to construct pedigrees that included the patients born in Clay County. Nine of 11 patients with IgAN born in Clay County, KY, USA were members of 1 or more of 5 pedigrees, each with 3 - 11 patients with IgAN. CONCLUSION: Our findings suggest the possibility of a low-penetrance ancestral mutation in the IgAN kindreds from Clay County.

Selective effects of tea extract and its phenolic compounds on human peripheral blood mononuclear cell cytokine secretions.

BACKGROUND: The effects of black tea extract (BTE) and some of its pure phenolics on human peripheral blood mononuclear cell (PBMC) cytokine secretion were examined in vitro. MATERIALS AND METHODS: The main steps of the study included chromatographic analysis of BTE and commercial green tea extract, Polyphenon 60, determination of the total antioxidant capacity of both the extracts and their phenolics, and finally evaluation of their effects on PBMCs. FINDINGS: Four major peaks in the chromatogram of BTE belonged to caffeine, gallic acid, epigallocatechin and epigallocatechin gallate, and the latter showed the highest antioxidant capacity. While pure phenolics at the concentration of 20 mM did not significantly affect PBMC cytokine secretion, BTE and Polyphenon 60 suppressed interleukin-1 beta, tumor necrosis factor-alpha and interleukin-6. Interestingly, the secretion of interferon-gamma after 24 h was slightly, but significantly, boosted by the extracts. CONCLUSION: BTE has selective pro-inflammatory cytokine-suppressing effects on human PBMCs.

Stress among medical students of Gorgan (South East of Caspian Sea), Iran.

OBJECTIVE: This study aims to estimate the prevalence of psychological stress and association between the levels of stress and study variables among Gorgan medical students. MATERIALS AND METHODS: All three year medical students (129 basic sciences students) in Gorgan Faculty of Medicine, Golestan University of Medical Sciences, were asked to complete the Kessler 10 questionnaire. RESULTS: The findings showed mild, moderate and severe stress among 26.22%, 20.50% and 14.75% study subjects. 39.35% of medical students had no stress. There was statistically significant association between year of study and stress levels (p= 0.040). CONCLUSION: The results indicate that there is a decrease in the psychological health of first year medical students. Provided that stress management courses are organised by medical schools, when the students arrive, they will cope up with the stress in coming years. These courses may reduce the negative effects of stress on medical students. By providing such courses and reducing stress level, medical students may improve their medical education.

Physiological dose of lycopene suppressed oxidative stress and enhanced serum levels of immunoglobulin M in patients with Type 2 diabetes mellitus: a possible role in the prevention of long-term complications.

OBJECTIVE: This study was undertaken to evaluate the antioxidant effects of lycopene in physiological doses and its possible effects on the immune response in patients with Type 2 diabetes mellitus (T2DM). RESEARCH DESIGN AND METHODS: A total of 35 patients with T2DM of both sexes aged 54+/-9 yr were enrolled in a double-blind placebo-controlled clinical trial conducted for 2 months. After a 2-week lycopene-free diet washout period, patients were allocated to either lycopene supplementation group (10 mg/day) (no.=16) or placebo group (no.=19), which were age- and sex matched. Patients were instructed to keep their diet and physical activity as unchanged as possible. RESULTS: While dietary intake of energy and body weight did not change, the ratio of serum total antioxidant capacity (TAC) to malondialdehyde (MDA) increased significantly in the lycopene group compared to the placebo group (p=0.007). Though a statistically significant increase in serum concentrations of lycopene (p<0.001) was not accompanied by enhanced delayed-type hypersensitivity response, a significant negative correlation was found between serum levels of lycopene and immunoglobulin (Ig)G (r=-0.338, p=0.008). Interestingly, variations of serum levels of lycopene directly correlated with those of IgM (r=0.466, p=0.005). There was an insignificant decrement in serum anti-oxidized LDL IgG levels in the lycopene group. CONCLUSIONS: Lycopene, probably by increasing TAC and inhibiting MDA-LDL formation, may attenuate T cell-dependent adaptive (pro-atherogenic) immune response. Meanwhile, with enhancement of innate immunity and hence prevention of ox-LDL uptake by macrophage and foam cell formation, lycopene may be effective in prevention of long-term diabetic complications, notably cardiovascular disease.

The opposite associations of lycopene and body fat mass with humoral immunity in type 2 diabetes mellitus: a possible role in atherogenesis.

This study examined the possible effects of lycopene at physiological dosage and body fat mass on the humoral immune response in patients with type 2 diabetes mellitus (T2DM). A total of 35 patients with Typ2 diabetes mellitus from both sexes aged 54+/-9 yrs from the Iranian Diabetes Society were introduced into a double blind placebo controlled clinical trial conducted for 2 months. After a 2-week lycopene free diet washout period, patients were allocated to either lycopene supplementation group (10mg/d) (n=16) or placebo age- and sex matched group (n=19) for 8 weeks. Patients were instructed to keep their diets and physical activities as unchanged as possible. Lycopene supplements increased serum lycopene levels (p<0.001). While intake of dietary energy and nutrients did not change in either groups, the ratio of total antioxidant capacity to malondialdehyde increased significantly in the lycopene group (p=0.007). There was an inverse correlation between serum levels of lycopene and those of IgG (r= -0.338, p=0.008). On the contrary, changes of serum levels of lycopene directly correlated with those of IgM (r=0.466, p=0.005). Interestingly, changes of the amount of fat mass correlated directly with those of serum IgG (r=0.415, p=0.044) but inversely with of serum IgM (r= -0.469, p=0.021). While truncal fat might promote adaptive humoral immunity, lycopene probably by inhibiting MDA-LDL formation might attenuate T cell dependent adaptive (pro-atherogenic) humoral immune response. These findings may have preventive implications in long term diabetic complications, notably atherogenesis.

Genes for diabetic nephropathy: sweet prospects on the horizon.

Familial aggregation of diabetic nephropathy has suggested an important role for hereditary factors in the development of this trait. Osterholm et al. provide further evidence that a gene on chromosome 3q confers susceptibility to this diabetic complication.

Determination of serum 25-hydroxy cholecalciferol using high-performance liquid chromatography: a reliable tool for assessment of vitamin D status.

This study was undertaken to design and set up a rather simple, reliable, and less expensive high-performance liquid chromatography (HPLC)-based method to assay 25(OH)D as a diagnostic tool for vitamin D assessment. Serum proteins were precipitated using ethanol and, after 10 minutes incubation at room temperature, methanol:isopropanol. The extraction was performed using hexane followed by evaporation under nitrogen flow. The sediment was then reconstituted in methanol and passed through a polypropylene filter. To run the chromatographic analysis, 20 microL of the filtrate was injected to the column. Peaks of 25(OH)D2 and 25(OH)D3 were both detected using a UV detector set at 265 nm. With a flow rate of 1.2 mL/minute, peaks of D3 and D2 vitamers were detected around 9.5 and 10.7 minutes, respectively. The intra- and inter-assay variations were 8.1% and 12.6%, respectively, and the recovery percent was found to be 100 +/- 5%. To compare the procedure with conventional methods, 90 serum samples from subjects (48 females and 42 males) aged 40.5 +/- 13.9 yrs, were analyzed for 25(OH)D using HPLC, competitive protein-binding assay (CPBA), and radioimmunoassay (RIA). Generally, CPBA and RIA assays both showed over-estimation of serum 25(OH)D, compared to HPLC. Though all three methods correlated significantly with each other, with the strongest between HPLC and RIA (r = 0.87, p < 0.001), both RIA and CPBA were found unreliable in detection of some deficient samples.

Selective microbiologic effects of tea extract on certain antibiotics against Escherichia coli in vitro.

OBJECTIVES: This study evaluated the microbiologic effects of black tea, compared to green tea, alone and in conjunction with selected antibiotics against Escherichia coli, the common cause of intestinal and urinary tract infections. DESIGN: This study was an in vitro evaluation of antibacterial effects of tea extracts. METHODS: Black and green tea extracts were analyzed by using high-performance liquid chromatography to compare their major polyphenol profiles. Different concentrations of the extracts or gallic acid (GA), the phenolic compound found with high concentration in the black tea extract, were employed for bacterial sensitivity tests, using pour plate and disc diffusion methods. The latter was used to evaluate the interactions between the extracts and certain anti-E. coli antibiotics. RESULTS: GA in black tea extract and epigallocatechin and epigallocatechin gallate in green tea extract are present in the highest concentrations, respectively. At concentrations of 25 mg/mL, both black and green teas after 5 and 7 hours completely inhibited E. coli growth. GA at concentrations of 5, 10, and 25 microg/mL after 7, 5 and 3 hrs, respectively, inhibited bacterial growth. Both black and green tea extracts had either synergistic or antagonistic effects at different concentrations on selected antibiotics, while GA showed a synergistic effect with all the antibiotics tested in a dose-dependent manner. The effect was more prominent with amikacin and sulfamethoxazole. CONCLUSIONS: The microbiologic effects of both black tea and green tea extracts on certain antibiotics against E. coli may vary, depending on the type of the tea extract (i.e., black vs. green), the amount of the extract, and the antibiotic being used.

Familial aggregation of primary glomerulonephritis in an Italian population isolate: Valtrompia study.

Hereditary factors are suspected to contribute to the pathogenesis of sporadic primary glomerulonephritis, but their contribution is difficult to delineate in the general population. We studied the prevalence of primary glomerulonephritis in an isolated population from the extreme northern Valtrompia valley, Northern Italy. Investigation of medical records, community urinary screening program and molecular characterization of the population's ancestry were performed; genealogies of affected individuals were researched. Forty-three patients with primary glomerulonephritis were identified: 25 had biopsy-proven disease (11 immunoglobulin A (IgA) nephropathy; eight mesangial proliferative glomerulonephritis without IgA deposits; four focal segmental glomerular sclerosis; two membranous nephropathy), and 18 had clinical glomerulonephritis. All 43 patients originated from three mountain villages (Collio, San Colombano, and Bovegno). In contrast, we found only four cases of primary glomerulonephritis in two nearby villages (Pezzaze and Tavernole) that shared similar population histories and lifestyles, demonstrating heterogeneity of risk factors for glomerulonephritis (P=3 x 10(-5)). All 43 affected individuals could be traced back to common ancestors (XVI-XVII centuries), enabling the construction of three large pedigree including three parent-child affected pairs and five affected siblings pairs. Molecular data showed lower genetic diversity and increased inbreeding in the Valtrompia population compared to the control population. Molecular and genealogical evidence of limited set of founders and the absence of shared nephritogenic environmental factors suggest that our patients share a common genetic susceptibility to the development of primary glomerulonephritis. Further molecular study of our families will offer the possibility to shed light on the genetic background underlying these glomerular disorders.

Intrauterine fetal death in mice caused by cytomegalovirus-derived peptide induced aPL antibodies.

Immunization of mice with beta2 glycoprotein I (beta2GPI) and also with GDKV, a synthetic peptide representing the phospholipid (PL)-binding site of beta2GPI, induced pathogenic aPL antibodies that bind and activate endothelial cells, enhanced thrombus formation and caused fetal death in pregnant mice. TIFI is a PL-binding peptide spanning the Thr101-Thr120 of ulb0-hcmva from human cytomegalovirus (CMV), which shares structural similarity with the PL-binding site of beta2GPI. Immunization with this peptide induced pathogenic aPL and anti-beta2GPI antibodies in mice. These antibodies activated endothelial cells and enhanced thrombus formation in vivo, but whether these antibodies cause fetal death in mice is not known. The objective of this study was to examine the effects of these antibodies on pregnancy outcome in mice. Two groups of pregnant BALB/c mice were injected with either hybridoma supernatant containing D3/AC10, a CMV-peptide-induced monoclonal aPL, at days four, eight and 12 of the pregnancy, 100 microg per mouse (study group) or with culture media alone (control group). The litter size was significantly smaller in the study group (4.80 +/- 1.15 versus 7.28 +/- 0.18, t = - 2.526, P < 0.03). In conclusion, aPL induced by CMV peptides may have pathogenic properties similar to human autoimmune aPL. These findings further support the hypothesis that at least in some patients with APS, pathogenic aPL antibodies may be generated by immunization with CMV products during incidental exposure to the virus via a molecular mimicry mechanism.