Comparative effects of β-carotene and fucoxanthin on retinol deficiency induced oxidative stress in rats

This study aimed at comparing antioxidant potential of fucoxanthin (FUCO) with β-carotene in relieving lipid peroxidation (Lpx) caused by retinol deficiency (RD) in rats. RD rats (n = 45) were fed a dose of either β-carotene (0.81 μmol) or FUCO (0.83 μmol). Plasma and liver lipid peroxide levels and activity of antioxidant enzymes catalase (CAT) and glutathione transferase (GST) were measured for 8 h. Results revealed that RD increased (P < 0.05) Lpx in plasma and liver by 34.3% and 19.4%, while the CAT activity in plasma (89%) and liver microsomes (91%) and GST in liver homogenate (31%) and liver microsomes (30%) were decreased (P < 0.05) compared to control (rats fed basal diet). FUCO suppressed (P < 0.05) the Lpx level by 7–85% (plasma) and 24–72% (liver) as compared to β-carotene (51–76%, 33–65%) over a period of 8 h. The activity of CAT in plasma and liver microsomes was higher (P < 0.05) in FUCO (90–95%, 85–93%) and β-carotene (87–96%, 79–91%) groups as compared to RD group. Similarly, the activity of GST in liver and its microsomes was also elevated (P < 0.05) in FUCO (44–51%, 22–51%) and β-carotene (19–54%, 30–43%) groups as compared to RD group. Results demonstrate that FUCO has greater potential than β-carotene in modulating Lpx, CAT, GST in plasma and liver of RD rats.     

Ghrelin is a Regulator of Cellular Apoptosis and Proliferation in the Rat Ovary

Apoptosis and proliferation are the common and essential events of reproductive function and development in the ovary, especially during follicular growth and atresia or luteal regression. Therefore, this study was set to investigate the influence of ghrelin treatment on apoptosis and proliferation specific indices in the rat ovary. Twenty-eight adult female Wistar rats were randomly allocated into control and treatment groups. Treatment group (n = 14) received 3 nmol of ghrelin as subcutaneous injection for 14 consecutive days or vehicle (normal saline) to the control rats. The animals from each group were equally sacrificed on days 9 and 14 after onset of ghrelin treatment and their ovaries were taken for immunohistochemical evaluation and caspase-3 assay. Accumulation of apoptosis-associated peptide Bax was significantly reduced following ghrelin treatment particularly in granulosa and luteal cells on day 14 (P < 0.01). In contrast, immunoreactivity against anti-apoptotic protein Bcl-2 was significantly elevated in ghrelin-exposed animals in granulosa, theca and luteal cells (P < 0.05). However, ghrelin administration was not able to change caspase-3 activity prominently, so that the means of enzyme activity were not statistically significant between groups (P > 0.05). Moreover, significant up-regulation of proliferation-associated peptide PCNA was also seen in the granulosa, theca and luteal cells of ghrelin-treated rats by day 14 (P < 0.05), but not on day 9. These findings indicate the first evidence of ghrelin involvement in the control of key gonadal functions, apoptosis and proliferation in the rat ovary, which is mainly mediated through decrease in Bax/Bcl-2 ratio consistent with upstream of PCNA level, however not depends on the reduction of caspase-3 activation. This may have potential implications that ghrelin can be considered as an apoptotic modulator of some ovarian disorders.     

A repertoire of biomarkers helps in detection and assessment of therapeutic response in epithelial ovarian cancer

In epithelial ovarian cancer (EOC), the cancer antigen 125 (CA-125) has been conventionally used to help in diagnosis and assessment of response to treatment. Currently, YKL-40 (Tyrosine–Lysine–Leucine-40) and circulating cell-free DNA are being evaluated for possession of similar ability. In this study, we aimed to assess the ability of a repertoire of potential biomarkers in detecting and assessing therapeutic response, in advanced EOC. Blood levels of CA-125, YKL-40, total cell-free DNA (CFDNA), cell-free nuclear DNA (CFnDNA), and cell-free mitochondrial DNA (CFmDNA) levels were measured in 100 untreated patients of advanced EOC from November 2009 to June 2011, and again on treatment completion from the 20 patients who appeared for follow-up analysis. Significantly, higher proportion of untreated patients had serum CA-125 >3 times upper limit of normal (ULN) (90.0 %; P < 0.0001) and plasma YKL-40 >ULN (77.0 %; P < 0.0001), both of which significantly decreased, Posttherapy. posttherapy, CFDNA (P < 0.0001), and CFnDNA (P < 0.0001) levels significantly decreased as compared to pretreatment levels. Positive and significant correlations existed between pretherapy CFDNA and CFnDNA [Spearman rho (ρ) = 1.000; P < 0.0001], and also with CFmDNA (ρ = 0.301; P = 0.002), separately between CFnDNA and CFmDNA (ρ = 0.303; P = 0.002), as well as between plasma YKL-40 and patient age (ρ = 0.353; (P < 0.0001). On treatment completion, CFDNA and CFnDNA levels showed positive and significant correlation (ρ = 1.000; P < 0.0001). Therefore serum CA-125 and plasma YKL-40 aid detection and assessment of therapeutic response, in advanced EOC. CFDNA and CFnDNA help in estimating extent of therapeutic response in advanced EOC.     

Structures and molecular forms of the ghrelin-family peptides

Ghrelin is an acylated peptide hormone produced mainly from the stomach. The major active products of the ghrelin gene in the stomach of rats, mice and humans are 28-amino acid peptides acylated at the serine-3 position with an n-octanoyl group (C8:0), called simply ghrelin. However, recent studies have revealed that the ghrelin gene can generate a variety of bioactive molecules besides ghrelin. These include acyl forms of ghrelin other than C8:0-ghrelin (i.e., n-decanoyl ghrelin or n-decenoyl ghrelin), des-acyl ghrelin, obestatin and ghrelin-associated peptides originated from the ghrelin gene. This review surveys the structures of the ghrelin peptides and molecular forms of ghrelin gene-derived products, and summarizes the knowledge about the functions of these peptides, with an emphasis on the acyl forms of the ghrelin peptide.     

Arterial Phase with CAIPIRINHA-Dixon-TWIST (CDT)–Volume-Interpolated Breath-Hold Examination (VIBE) in Detecting Hepatic Metastases

PURPOSE: To evaluate lesion enhancement performance of Multi-Arterial CAIPIRINHA-Dixon-TWIST–Volume-Interpolated Breath-Hold Examination (MA-CDT-VIBE) for the detection of hepatic metastases. MATERIALS AND METHODS: Thirty-one patients with suspicious hepatic metastases were enrolled in this retrospective study. Two independent radiologists scored visualization of each lesion on a scale of 1 (poor visualization) to 11 (excellent visualization) on 11 sets of images. These included 6 hepatic arterial sub-phases acquired in one breath-hold, 1 series of the mean of 6 hepatic arterial sub-phases, 3 subtracted arterial sub-phases, and 1 portal venous phase. The phases with good (score 8–10) and excellent (score 11) lesion visualization were identified, and the number of lesions seen on each of these phases was compared to the number of lesions that was seen best on the equivalent-to-conventional single arterial phase as well as to those that were see best on the mean of 6 hepatic arterial sub-phases. Inter-reader agreement was also calculated. RESULTS: The MA-CDT-VIBE was successfully acquired in 25 patients with hypervascular metastases (96 lesions) and 6 patients with hypovascular metastases (13 lesions). In case of hypervascular metastases, the 6th/6 arterial sub-phase had excellent lesion visualization (sore of 11) in 56 and 44 lesions for the 2 readers, respectively. Good lesion visualization (score of 8-10) was recorded in 5th/6 arterial subphases, in 81 and 67 lesions for the 2 readers, respectively. In case of hypovascular metastases, the portal venous phase had excellent lesion visualization (sore of 11) in all 13 lesions for the 2 readers. Good lesion visualization (score of 8–10) was recorded in 12 and 13 lesions on the 5th/6 and 6th/6 arterial subphases, respectively. More hypervascular lesions scored good (score of 8–10) and excellent (score of 11) on the 5th/6 and 6th/6 phases of MA-CDT-VIBE compared with the equivalent-to-conventional single arterial phase (3rd/6) and the set with mean of 6 hepatic arterial sub-phases. The results were statistically significant (t test, P < .0001). Inter-reader agreement was good for hypervascular lesions (kappa = 0.627, P < .0001) and excellent for hypovascular lesions (kappa = 1.0, P < .0001), respectively. CONCLUSIONS: The MA-CDT-VIBE improves lesion conspicuity by providing a wide observation window for hypervascular lesions. For hypovascular lesions, the advantage of multiple arterial sub-phases over the portal venous phase is not apparent.     

Circulating Obestatin Levels Correlate with Fasting Insulin and HOMA-IR but Not with Hypertension in Elderly Men

Obestatin, encoded by the same gene as ghrelin, was first described as a physiological opponent of ghrelin. The association between circulating obestatin levels and blood pressure remains unclear. Furthermore, adequate information is non-existent regarding the older male population with hypertension. For this purpose, we enrolled 185 unrelated hypertensive male patients aged ≥80 years (range 80–102 years). One hundred seventy nine age-matched healthy subjects served as controls. Plasma levels of obestatin and insulin were measured using commercial ELISA and RIA. HOMA-IR was calculated using standard method. We found that plasma obestatin levels correlated significantly with insulin levels (P = 0.034) and homeostasis model assessment index for insulin resistance (HOMA-IR: P = 0.028). However, plasma obestatin differed non-significantly between hypertensive (5.06 ± 0.68 ng/mL) and non-hypertensive (4.72 ± 0.82 ng/mL) individuals. Plasma obestatin levels were not associated with systolic (P = 0.818) or diastolic (P = 0.564) blood pressure, waist-to-hip ratio (WHR: P = 0.725), uric acid (P = 0.603), total cholesterol (TC: P = 0.589), low-density lipoprotein cholesterol (LDL-C: P = 0.057); high-density lipoprotein cholesterol (HDL-C: P = 0.432), triglyceride (TG: P = 0.418), and fasting blood glucose (FBG: P = 0.101). We, therefore, concluded that fasting circulating obestatin levels did not directly correlate with blood pressure in men aged ≥80 years.     

Is high prevalence of vitamin D deficiency a correlate for attention deficit hyperactivity disorder?

The aim of the study was to determine the association between vitamin D and attention deficit hyperactivity disorder (ADHD), and difference in the level of vitamin D in ADHD children and control. This a case–control study carried out in school health and primary health care clinics. A total of 1,331 children and adolescents who were diagnosed with ADHD based on clinical criteria and standardized questionnaires were enrolled in this study and were matched with 1,331 controls, aged 5–18 years old. Data on body mass index (BMI), clinical biochemistry variables including serum 25-hydroxyvitamin D were collected. The study found significant association between ADHD and vitamin D deficiency after adjusting for BMI and sex (adj. OR 1.54; 95 % CI 1.32–1.81; P < 0.001). Majority of the ADHD children were in the age group 5–10 years (40.7 %), followed by 11–13 years (38.4 %). The proportion of BMI <85th percentile was significantly over represented in ADHD group as compared to healthy control (87.8 vs. 83 %; P < 0.001, respectively), while on the other hand, BMI >95th percentile was over represented in the control than ADHD group (7.6 vs. 4.6 %; P < 0.001, respectively). Mean values of vitamin D (ng/mL) were significantly lower in ADHD children (16.6 ± 7.8) than in healthy children (23.5 ± 9.0) (P < 0.001). There was significant correlation between vitamin D deficiency and age (r = ?0.191, P = 0.001); calcium (r = 0.272, P = 0.001); phosphorous (r = 0.284, P = 0.001); magnesium (r = 0.292, P = 0.001); and BMI (r = 0.498, P = 0.001) in ADHD children. The vitamin D deficiency was higher in ADHD children compared to healthy children.     

Preliminary findings on the influence of FTO rs9939609 and MC4R rs17782313 polymorphisms on resting energy expenditure,leptin and thyrotropin levels in obese non-morbid premenopausal women

Given that leptin, ghrelin and thyrotropin play a major role in the regulation of resting energy expenditure (REE) and that the FTO rs9939609 and the MC4R rs17782313 polymorphisms have been proposed to affect energy homeostasis, we hypothesized that both polymorphisms are associated with REE and that these relationships can be mediated by leptin, ghrelin and thyrotropin in obesity. Therefore, the present study aimed to examine the relationships between FTO rs9939609 and the MC4R rs17782313 with REE, leptin, ghrelin and thyrotropin levels in obese women. The study comprised 77 obese (body mass index 34.0?±?2.8 kg/m²) women (age 36.7?±?7 years). We measured body composition by dual-energy X-ray absorptiometry and REE by indirect calorimetry. We analysed fasting leptin, ghrelin and thyrotropin levels and the ratio of leptin to fat mass was calculated. Genotype distributions of the polymorphisms did not deviate from Hardy–Weinberg expectations (P values >0.2). Women carrying the A allele of the FTO rs9939609 had lower REE (1,580?±?22 vs. 1,739?±?35 kcal/day, P?<?0.001) and higher leptin to fat mass ratio (1.33?±?0.05 vs. 1.13?±?0.08 ng/ml kg, P?<?0.05) and thyrotropin levels (1.93?±?0.10 vs. 1.53?±?0.16 μU/ml, P?<?0.05) regardless of age and body mass index. We found no significant influence of the MC4R rs17782313 on energy metabolism or biochemical variables. Our findings confirm that the A allele of the FTO rs9939609 is associated with lower REE and increased plasma leptin levels. We also found an association between the FTO rs9939609 and thyrotropin, suggesting the possible influence of FTO in the hypothalamic–pituitary–thyroid axis as a potential mechanism of the increased adiposity.     

ELISA examining urinary angiotensinogen as a potential indicator of intrarenal renin–angiotensin system (RAS) activity: a clinical study of 128 chronic kidney disease patients

In the current study, we measured urinary angiotensinogen (AGT) through enzyme-linked immunoadsordent assay (ELISA) and analyzed its correlation with intrarenal renin–angiotensin system (RAS) activity in 128 chronic kidney disease (CKD) patients. Urinary and plasma renin activity, AGT, angiotensin II (Ang II) and aldosterone levels were also measured by radioimmunoassay (RIA) or ELISA in these participants. Further, the expression level of intrarenal renin, AGT, Ang II and Ang II receptors were examined by immunohistochemistry staining (IHCS) in 72 CKD patients. Their correlations with urinary AGT were also analyzed. We found that the urinary AGT level was positively correlated with hypertension (ρ = 0.28, P < 0.01), urinary protein (r = 0.38, P < 0.01), urinary Ang II (r = 0.29, P < 0.05), urinary type IV collagen (Col IV) (r = 0.56, P < 0.01), and was negatively correlated with estimated glomerular filtration rate (eGFR) (r = ?0.28, P < 0.01), urinary sodium (r = ?0.22, P < 0.05) and serum AGT (r = ?0.27, P < 0.01). Multiple regression analysis indicated low serum AGT (P < 0.01), high urinary protein (P < 0.01), high urinary Ang II (P < 0.05) and high urinary Col IV (P < 0.01) were correlated significantly with high urinary AGT. Urinary AGT level was positively correlated with intrarenal expression level of AGT (ρ = 0.46, P < 0.01), Ang II (ρ = 0.56, P < 0.01) and Ang II type 1 receptor (ρ = 0.32, P < 0.01), as detected by IHCS. Together, these data suggest that urinary AGT might be a potential biomarker of intrarenal RAS and Ang II activities in CKD patients.     

Oxidative Stress-Related Genes in Type 2 Diabetes: Association Analysis and Their Clinical Impact

Worldwide prevalence of diabetes mellitus motivates a number of association studies to be conducted throughout the world. Eleven polymorphisms from nine candidate genes in oxidative stress pathway have been analyzed in eastern Indian type 2 diabetic patients (n = 145) and healthy controls (n = 100). Different biochemical parameters were also analyzed for their association with the disease. Significant associations were observed for rs2070424 A>G SOD1 (OR 3.91, 95% CI 2.265–8.142, P < 0.001), rs854573 A>G PON1 (OR 3.415, 95% CI 2.116–5.512, P < 0.001), rs6954345 G>C PON2 (OR 3.208, 95% CI 2.071–4.969, P < 0.001), RAGE rs1800624 ?374 T>A (OR 3.58, 95% CI 2.218–5.766, P < 0.001), and NOS3 ?786 T>C (OR 3.75, 95% CI 2.225–6.666, P < 0.001). Haplotype containing two risk alleles of PON1 and PON2 genes was significantly associated with disease (OR 8.34, 95% CI 1.554–44.804, P < 0.002). Our results suggest that carriers of major and efficient alleles of oxidative stress genes are more likely to survive the comorbid complications and single copy of risk allele is sufficient for developing the disease.     

Relationship between atomoxetine plasma concentration, treatment response and tolerability in attention-deficit/hyperactivity disorder and comorbid oppositional defiant disorder

The purpose of this study was to examine whether atomoxetine plasma concentration predicts attention-deficit/hyperactivity disorder (ADHD) or oppositional defiant disorder (ODD) response. This post-hoc analysis assessed the relationship between atomoxetine plasma concentration and ADHD and ODD symptoms in patients (with ADHD and comorbid ODD) aged 6–12 years. Patients were randomly assigned to atomoxetine 1.2 mg/kg/day (n = 156) or placebo (n = 70) for 8 weeks (Study Period II). At the end of 8 weeks, ODD non-remitters (score >9 on the SNAP-IV ODD subscale and CGI-I > 2) with atomoxetine plasma concentration <800 ng/ml at 2 weeks were re-randomized to either atomoxetine 1.2 mg/kg/day or 2.4 mg/kg/day for an additional 4 weeks (Study Period III). ODD remitters and non-remitters with plasma atomoxetine ≥800 ng/ml remained on 1.2 mg/kg/day atomoxetine for 4 weeks. Patients who received atomoxetine, completed Study Period II, and entered Study Period III were included in these analyses. All the groups demonstrated improvement on the SNAP-IV ODD and ADHD-combined subscales (P < .001). At the end of Study Periods II and III, ODD and ADHD improvement was significantly greater in the remitter group compared with the non-remitter groups. Symptom improvement was numerically greater in the non-remitter (2.4 mg/kg/day compared with the non-remitter 1.2 mg/kg/day) group. Atomoxetine plasma concentration was not indicative of ODD and ADHD improvement after 12 weeks of treatment. ADHD and ODD symptoms improved in all the groups with longer duration on atomoxetine. Results suggest atomoxetine plasma concentration does not predict ODD and ADHD symptom improvement. However, a higher atomoxetine dose may benefit some patients.     

Oxidized Low Density Lipoprotein and Inflammation in Gout Patients

To analyze the levels of oxidized low density lipoprotein (ox-LDL) and inflammatory cytokines in the plasma of gout patients. The levels of ox-LDL, hypersensitive C-reactive protein (hs-CRP), interleukin-1β, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured in the plasma of 41 gout patients [28 in acute phase episode, 13 in intermittent phase (IP)], and in 40 healthy controls. The relationship between ox-LDL and inflammation was also explored by measuring the levels of several pro-inflammatory cytokines in the plasma. The plasma levels of ox-LDL, hs-CRP, IL-6 and TNF-α were significantly increased in patients with gout in the acute phase compared to those in the IP group and healthy controls (P < 0.05), but the levels of TGF-β were significantly lower in the acute phase group than in the IP group and healthy controls (P < 0.01). The levels of ox-LDL in the gout patients in the IP were significantly higher than those in healthy controls (P < 0.05). Correlation analysis indicated that the levels of ox-LDL were positively correlated with hs-CRP, IL-6 and TNF-α (r = 0.343, r = 0.386, r = 0.659, P < 0.01, respectively), but negatively correlated with TGF-β levels in patients in the acute phase (r = ?0.240, P < 0.05). The levels of ox-LDL in gout patients were significantly higher than those in healthy controls. The changes in ox-LDL levels may be associated with enhanced inflammation in gout patients.     

Age-related and seasonal variations in plasma uncarboxylated osteocalcin in male Murrah buffaloes

The present study aims to investigate the age-related and seasonal variations in plasma uncarboxylated osteocalcin (UcOCN) in male Murrah buffaloes. In experiment 1, body weight, body dimensions and plasma UcOCN levels were estimated in 120 male Murrah buffaloes aged between 18 days and 8 years. We observed a significant (p < 0.0001) decrease in plasma UcOCN with age and their levels ranged between 0.25 and 9.25 ng/ml. In experiment 2, the plasma UcOCN, cortisol, triiodothyronine (T₃), thyroxine (T₄) and prolactin levels were determined during spring and summer in 6–34 months (n = 24) old male Murrah buffaloes. Significant (p < 0.0001) higher plasma UcOCN levels were observed in spring than in summer. The plasma T₃ and T₄ levels were positively correlated with plasma UcOCN, whereas the plasma cortisol and prolactin were negatively correlated. Further, to observe the effect of season on temporal variations of UcOCN, Murrah buffaloes aged between 4–6 (n = 5), 14–16 (n = 5) and 28–30 (n = 5) months were bled at an interval of 30 min, between 0700 and 01030 h, during both the seasons. We observed significant effects of season (p < 0.0001), age (p < 0.0001) and season-by-age interaction (p < 0.01) on plasma UcOCN. The reasons for variations in plasma UcOCN with age and season will be discussed.     

Effects of rumen-protected methionine on plasma amino acid concentrations during a period of weight loss for late gestating beef heifers

This study determined changes in plasma amino acid concentration in late-gestating (beginning 58 ± 1.02 days prior to calving), primiparous, winter-grazing range heifers receiving wheat middling-based supplement without (CON) or with rumen-protected methionine (MET) to provide 15 g dl-MET each day. Plasma was collected on days ?2 and 0 (start of MET supplementation just prior to individually receiving supplement at 0700 hours). Plasma was sampled again on days 40, 42 and 44 prior to supplementation at 0700 and 1100 hours (4 h after receiving daily supplement). Data were analyzed with cow as the experimental unit. Continuous variables were analyzed by the main effects of treatment, date, or time and their interaction when appropriate. Comparable BW (P = 0.32) and BCS (P = 0.83) over the 44-day metabolism trial were found between both CON- and MET-fed heifers. MET-supplemented heifers had greater (P < 0.01) plasma concentrations of methionine indicating that the rumen-protection technology successfully delivered methionine to the small intestine. Supplementation with rumen-protected dl-MET caused a significant supplement × date interaction for glutamine (P = 0.03), glycine (P = 0.02), methionine (P < 0.01), and serine (P = 0.05). In addition, trends for supplement × date interactions were detected for leucine (P = 0.07), threonine (P = 0.09), valine (P = 0.08), total amino acids (TAA; P = 0.08), non essential amino acids (NEAA; P = 0.08), branched chain amino acids (BCAA; P = 0.08), and glucogenic amino acids (GLUCO; P = 0.08). These results suggest that the BCAA (leucine and valine) were utilized more efficiently with MET supplemented heifers compared to CON supplemented heifers. Plasma AA concentrations for glutamic acid (P < 0.01), histidine (P = 0.01), tyrosine (P < 0.01), and EAA (P < 0.01), all decreased throughout the study. These results further confirm methionine is a limiting amino acid in forage fed late-gestating heifers and further suggests the limitation when grazing dormant range forages as shown by improved utilization of other plasma amino acids when supplemental methionine was provided.     

Gas chromatography–mass spectrometry analysis of fatty acid profiles of Antarctic and non-Antarctic yeasts

The fatty acid profiles of Antarctic (n = 7) and non-Antarctic yeasts (n = 7) grown at different temperatures were analysed by gas chromatography–mass spectrometry. The Antarctic yeasts were enriched in oleic 18:1 (20–60 %), linoleic 18:2 (20–50 %) and linolenic 18:3 (5–40 %) acids with lesser amounts of palmitic 16:0 (<15 %) and palmitoleic 16:1 (<10 %) acids. The non-Antarctic yeasts (n = 4) were enriched in 18:1 (20–55 %, with R. mucilaginosa at 75–80 %) and 18:2 (10–40 %) with lesser amounts of 16:0 (<20 %), 16:1 (<20 %) and stearic 18:0 (<10 %) acids. By contrast, Saccharomyces cerevisiae strains (n = 3) were enriched in 16:1 (30–50 %) and 18:1 (20–40 %) with lesser amounts of 16:0 (10–25 %) and 18:0 (5–10 %) acids. Principal component analysis grouped the yeasts into three clusters, one belonging to the S. cerevisiae strains (enriched in 16:0, 16:1 and 18:1), one to the other non-Antarctic yeasts (enriched in 18:1 and 18:2) and the third to the Antarctic yeasts (enriched in 18:2 and 18:3).     

Genetic association of IDE,POU2F1, PON1, IL1α and IL1β with type 2 diabetes in Pakistani population

A number of genes are known to be involved in glucose homeostasis. Mutations and polymorphisms in candidate genes may effect insulin production, action or resistance. This study was designed to report the association of genetic polymorphism with the type 2 diabetes (T2D) in Pakistani population. A total of 458 subjects (case n = 288, control n = 170) participated in the study. Nine single nucleotide polymorphisms were investigated in genes IDE (rs6583813 C>T, rs7910977 C>T), POU2F1 (rs3767434 A>T, rs10918682 A>T, rs2146727 A>G), WFS1 (rs734312 A>G), PON1 (rs854560 T>A), IL1α (rs1800587 C>T) and IL1β (rs1143634 C>T). Genotyping was performed by DNA sequencing after nested polymerase chain reaction of targeted regions. Results indicated that rs7910977 in IDE showed significant association with the development of T2D [P = 0.012, OR 1.677 (95 % CI 1.112–2.438)]. The rs10918682 in POU2F1 was associated with T2D [P < 0.001, OR 3.606 (95 % CI 2.165–6.005)]. The rs854560 in PON1was associated with incidences of T2D and increased the risk of cardiovascular complications [P = 0.031, OR 0.663 (95 % CI 0.455–0.965)] in diabetics. The rs734312 from WFS1 gene was associated with diabetes at genotype level (P < 0.01). Haplotype analysis of rs1800587–rs1143634 depicted CC haplotype increased the susceptibility to diabetes (P < 0.05). Haplotype GAA from rs2146727–10918682–rs3767434 was protective against diabetes (P < 0.01) and GGA exhibited the association with T2D (P < 0.01). Haplotype CT from rs6583813–rs7910977 was protective against diabetes (P = 0.02). Our study provided evidence to IDE, PON1, WFS1, POU2F1, IL1α and IL1β associated with T2D in Pakistanis.     

Expression of “brown-in-white” adipocyte biomarkers shows gender differences and the influence of early dietary exposure

Fish oil supplementation provides an inconsistent degree of protection from cardiovascular disease (CVD), which may be attributed to genetic variation. Single nucleotide polymorphisms (SNPs) in the elongation-of-very-long-chain-fatty-acids-2 (ELOVL2) gene have been strongly associated with plasma proportions of n-3 long-chain polyunsaturated fatty acids (LC-PUFA). We investigated the effect of genotype interaction with fish oil dosage on plasma n-3 LC-PUFA proportions in a parallel double-blind controlled trial, involving 367 subjects randomised to treatment with 0.45, 0.9 and 1.8 g/day eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (1.51:1) or olive oil placebo for 6 months. We genotyped 310 subjects for ELOVL2 gene SNPs rs3734398, rs2236212 and rs953413. At baseline, carriers of all minor alleles had lower proportions of plasma DHA than non-carriers (P = 0.021–0.030). Interaction between genotype and treatment was a significant determinant of plasma EPA (P < 0.0001) and DHA (P = 0.004–0.032). After the 1.8 g/day dose, carriers of ELOVL2 SNP minor alleles had approximately 30 % higher proportions of EPA (P = 0.002–0.004) and 9 % higher DHA (P = 0.013–0.017) than non-carriers. Minor allele carriers could therefore particularly benefit from a high intake of EPA and DHA in maintaining high levels of plasma n-3 PUFA conducive to protection from CVD.     

Adaptive developmental plasticity in methylene tetrahydrofolate reductase (MTHFR) C677T polymorphism limits its frequency in South Indians

Methylene tetrahydrofolate reductase (MTHFR) C677T polymorphism shows considerable heterogeneity in its distribution in humans worldwide. The current study was conducted to investigate whether this polymorphism exhibited adaptive developmental plasticity in the control of the TT-genotype frequency. We screened 1,818 South Indian subjects (895 males and 923 females) for MTHFR C677T polymorphism using PCR–restriction fragment length polymorphism approach. MTHFR 677T-allele frequency in males and females was 9.1 and 11.0 %, respectively. Compared to females, males had lower frequency of TT-genotype [odds ratio 0.31, 95 % confidence interval (CI) 0.08–1.01]. The frequency of MTHFR 677T-allele was highest in the age group of 20–40 years and it gradually decreased from 40–60 to 60–80 years (P _trend <0.0001). MTHFR 677TT-genotype was associated with 7.02-folds (95 % CI: 2.12–25.63, P < 0.0001) cumulative risk for recurrent pregnancy loss (RPL), neural tube defects (NTDs) and deep vein thrombosis (DVT). Linear regression model suggested that male gender exhibited increased homocysteine levels by 9.35 μmol/L while each MTHFR 677T-allele contributed to 4.63 μmol/L increase in homocysteine. Plasma homocysteine showed inverse correlation with dietary folate (r = ?0.17, P < 0.0001), B₂ (r = ?0.14, P < 0.0001) and B₆ (r = ?0.07, P = 0.03). Examination of the spontaneously aborted fetuses (n = 35) showed no significant association of fetal genotype on its in utero viability. From the current study, it was concluded that C677T seemed to have acquired adaptive developmental plasticity among South Indians due to environmental influences thus contributing to hyperhomocysteinemia and its associated complications such as RPL, NTDs, DVT, etc.     

Natural iron isotopic composition of blood is an indicator of dietary iron absorption efficiency in humans

Persistent impairments in the regulation of intestinal iron absorption result in iron deficiency or iron accumulation in the long term. Diagnosis remains difficult unless pathological symptoms develop as iron absorption varies strongly between meals and days. Variations in the natural iron isotopic composition of whole blood have recently been suggested as a novel parameter to assess long-term differences in intestinal absorption efficiency between individuals. In this study, baseline blood samples collected in two previous conventional iron absorption studies in Swiss and Thai women using stable isotope tracers were reanalyzed by multicollector inductively coupled plasma mass spectrometry. The natural iron isotopic compositions obtained were compared with fractional absorption from the test meals observed in these earlier trials. Correlations of natural blood iron isotopic composition and fractional absorption from the test meals were found to be highly significant in both cohorts (for Swiss women, r = 0.40, P = 0.01, n = 38; for Thai women, r = 0.57, P < 0.01, n = 24), with the blood of both ethnicities clearly differing in iron isotopic composition (P < 0.001). Combining the findings of this study and those of recent animal and human studies confirms that blood iron isotopic patterns may serve as a novel compound biomarker of iron metabolism to assess impairments in regulation of intestinal iron absorption in individuals or population groups.     

Short-term supplementation with flavanol-rich cocoa improves lipid profile,antioxidant status and positively influences the AA/EPA ratio in healthy subjects

We evaluated the short-term effects of a flavanol-rich cocoa (FRC) on lipid profile and selected oxidative stress biomarkers such as oxidized low-density lipoprotein (oxLDL), glutathione (GSH), and F₂-isoprostane. We also assessed whether FRC modulates plasma levels of polyunsaturated fatty acids (PUFA) in healthy individuals. The subjects (n=48) were randomly assigned to a low-cocoa group (1 g/d; ~55 mg flavanols) (n=16), middle-cocoa group (2 g/d; ~110 mg flavanols) (n=16), or a high-cocoa group (4 g/d; ~220 mg flavanols) (n=16). The samples were collected at baseline, at 1, 2, and 4 h post initial consumption of FRC, and after 4 weeks of FRC supplementation. The peak plasma concentration of (−)-epicatechin metabolites reached a maximum level (578±61 nM; P<.05) at 2 h after ingestion of FRC. After 4 weeks, total cholesterol (−12.37±6.63; P<.0001), triglycerides (−3.81±2.45; P<.0001), plasma LDL (−14.98±6.77; P<.0001), and oxLDL (−95.61±41.69; P<.0001) decreased in the high-cocoa group, compared with baseline. We also found that plasma high-density lipoprotein (HDL) (+3.37±2.06; P<.0001) concentrations increased significantly in the same group. Total GSH significantly increased in all FRC-treated groups (+209.73±146.8; P<.0001), while urinary F₂-isoprostane levels decreased in the middle- (−0.73±0.16; P<.0001) and high-cocoa (−1.62±0.61; P<.0001) groups. At the end of the four-week study, a significant reduction of arachidonic acid (AA)/eicosapentaenoic acid (EPA) ratio was observed in the low-(−2.62±2.93; P=.003), middle- (−5.24±2.75; P<.0001) and high-cocoa (−7.76±4.96; P<.0001) groups, compared with baseline. Despite the small sample size used in this study, these data extend previous clinical and experimental studies, providing new insights into the health benefits of cocoa flavanols.