Evaluation of D-loop hypervariable region I variations, haplogroups and copy number of mitochondrial DNA in Bangladeshi population with type 2 diabetes.

The profound impact of mitochondrion in cellular metabolism has been well documented. Since type 2 diabetes (T2D) is a metabolic disorder, mitochondrial dysfunction is intricately linked with the disease pathogenesis. Mitochondrial DNA (mtDNA) variants are involved with functional dysfunction of mitochondrion and play a pivotal role in the susceptibility to T2D. In this study, we opted to find the association of mtDNA variants within the D-loop hypervariable region I (HVI), haplogroups and mtDNA copy number with T2D in Bangladeshi population. A total of 300 unrelated Bangladeshi individuals (150 healthy and 150 patients with T2D) were recruited in the present study, their HVI regions were amplified and sequenced using Sanger chemistry. Haplogrep2 and Phylotree17 tools were employed to determine the haplogroups. MtDNA copy number was measured using primers of mitochondrial tRNALeu (UUR) gene and nuclear ß2-microglobulin gene. Variants G16048A (OR:0.12, p = 0.04) and G16129A (OR: 0.42, p = 0.007) were found to confer protective role against T2D according to logistic regression analysis. However along with G16129A, two new variants C16294T and T16325C demonstrated protective role against T2D when age and gender were adjusted. Haplogroups A and H showed significant association with the risk of T2D after adjustments out of total 19 major haplogroups identified. The mtDNA copy numbers were stratified into 4 groups according to the quartiles (groups with lower, medium, upper and higher mtDNA copy numbers were respectively designated as LCN, MCN, UCN and HCN). Patients with T2D had significantly lower mtDNA copy number compared to their healthy counterparts in HCN group. Moreover, six mtDNA variants were significantly associated with mtDNA copy number in the participants. Thus, our study confers that certain haplogroups and novel variants of mtDNA are significantly associated with T2D while decreased mtDNA copy number (though not significant) has been observed in patients with T2D. However, largescale studies are warranted to establish association of novel variants and haplogroup with type 2 diabetes.

Fc-gamma IIIa-V158F receptor polymorphism contributes to the severity of Guillain-Barré syndrome.

OBJECTIVE: Guillain-Barré syndrome (GBS) is a rare, life-threatening disorder of the peripheral nervous system. Immunoglobulin G Fc-gamma receptors (FcγRs) mediate and regulate diverse effector functions and are involved in the pathogenesis of GBS. We investigated whether the FcγR polymorphisms FcγRIIa H/R131 (rs1801274), FcγRIIIa V/F158 (rs396991), and FcγRIIIb NA1/NA2, and their haplotype patterns affect the affinity of IgG-FcγR interactivity and influence GBS susceptibility and severity. METHODS: We determined FcγR polymorphisms in 303 patients with GBS and 302 ethnically matched healthy individuals from Bangladesh by allele-specific polymerase chain reaction. Pairwise linkage disequilibrium and haplotype patterns were analyzed based on D ́statistics and the genotype package of R statistics, respectively. Logistic regression analysis and Fisher's exact test with corrected P (Pc) values were employed for statistical comparisons. RESULTS: FcγRIIIa-V158F was associated with the severe form of GBS compared to the mild form (P = 0.005, OR = 2.24, 95% CI = 1.28-3.91; Pc = 0.015); however, FcγR genotypes and haplotype patterns did not show any association with GBS susceptibility compared to healthy controls. FcγRIIIa-V/V158 and FcγRIIIb-NA2/2 were associated with recent Campylobacter jejuni infection (P ≤ 0.001, OR = 0.36, 95% CI = 0.23-0.56; Pc ≤ 0.003 and P = 0.004, OR = 1.70, 95% CI = 1.18-2.44; Pc ≤ 0.012, respectively). Haplotype 1 (FcγRIIa-H131R- FcγRIIIa-V158F- FcγRIIIb-NA1/2) and the FcγRIIIb-NA2/2 genotype were more prevalent among anti-GM1 antibody-positive patients (P = 0.031, OR = 9.61, 95% CI = 1.24-74.77, Pc = 0.279; P = 0.027, OR = 1.62, 95% CI = 1.06-2.5, Pc = 0.081, respectively). INTERPRETATION: FcγR polymorphisms and haplotypes are not associated with susceptibility to GBS, though the FcγRIIIa-V158F genotype is associated with the severity of GBS.

Association of matrix metalloproteinase-9 polymorphism with severity of Guillain-Barré syndrome.

Guillain-Barré syndrome (GBS) is an immune-mediated neurological disorder with a multifaceted nature. Infectious agents and immune-response genetic host factors may contribute to the development of GBS. The matrix metalloproteinase-9 (MMP-9), an enzyme is upregulated by pro-inflammatory cytokines and might play an important role in the pathogenesis of GBS. This study investigated the association of a single nucleotide polymorphism (-1562C/T, rs3918242) in the MMP9 gene with the susceptibility and severity of GBS in Bangladesh. The allele and genotype distributions of the MMP9 polymorphism were not significantly different between 303 patients with GBS and 303 healthy controls. Serum concentrations of MMP-9 were significantly elevated in patients with GBS compared to healthy controls (P ≤.0001). No significant association of MMP-9 (-1562C/T) polymorphism was observed with disease prognosis. The frequencies of the MMP9-1562 CT genotype and T allele (P = .01, OR = 2.28, 95% CI = 1.22-4.22; Pc = 0.03 and P = .012, OR = 2.0, 95% CI = 1.14-3.38; Pc = 0.024, respectively) were significantly increased in patients with severe form of GBS, indicates the MMP9 polymorphism plays a role in the disease severity of GBS.

Human leukocyte antigen-DQB1 polymorphisms and haplotype patterns in Guillain-Barré syndrome.

OBJECTIVE: The etiology of Guillain-Barré syndrome (GBS) remains enigmatic, although genetic and environmental factors are speculated to be associated with this autoimmune condition. We investigated whether polymorphisms and the haplotype structures of the human leukocyte antigen (HLA)-DQB1 gene relate to the autoimmune response to infection and affect the development of GBS. METHODS: HLA-DQB1 polymorphic alleles (*0201, *030x, *0401, *050x, *060x) were determined for 151 Bangladeshi patients with GBS and 151 ethnically matched healthy controls using sequence-specific polymerase chain reaction. Pairwise linkage disequilibrium (LD) and haplotype patterns were analyzed based on D ́statistics and the genotype package in R statistics, respectively. Association studies were conducted using Fisher's exact test and logistic regression analysis. The Bonferroni method was applied to correct for multiple comparisons, whereby the P-value was multiplied with the number of comparisons and denoted as Pc (Pc, P corrected). RESULTS: No associations were observed between HLA-DQB1 alleles and susceptibility to disease in the comparison between GBS patients and healthy subjects. Haplotype 9 (DQB1*0303-*0601) tended to be less frequent among patients with GBS than healthy controls (P = 0.006, OR = 0.49, 95% CI = 0.30-0.82; Pc = 0.06). Haplotype 5 (DQB1*0501-*0602) and the DQB1*0201 alleles were more frequent in the Campylobacter jejuni-triggered axonal variant of GBS (P = 0.024, OR = 4.06, 95% CI = 1.25-13.18; Pc = 0.24) and demyelinating subtype (P = 0.027, OR = 2.68, 95% CI = 1.17-6.17; Pc = 0.35), though these associations were not significant after Bonferroni correction. INTERPRETATION: This study indicates that HLA-DQB1 polymorphisms are not associated with susceptibility to GBS. In addition, these genetic markers did not influence the clinical features or serological subgroup in patients with C. jejuni-triggered axonal variant of GBS.

Hyperphosphataemia is associated with the diabetes-related cardiovascular risk factors.

The serum phosphorus level is recently considered as one of the foretelling markers for the severity of cardiovascular diseases (CVD). We therefore investigated whether the serum phosphorus level in the diabetic patients against healthy individuals could act as a possible marker for identification of vulnerability to cardiovascular disease. One hundred and thirty two human subjects were involved in the study among which one hundred and four subjects are CVD patients and twenty eight were healthy individuals. The levels of the lipid profile and the glycemic status were significantly increased in the patients than those of the control subjects (Fasting glucose, FS=8.3 ± 0.3 vs. 6.1 ± 0.0 mmol/L; blood glucose 2 h after breakfast (STAB)=12.0 ± 0.5 vs. 8.5 ± 0.7, mmol/L; HbA1c (%),6.7 ± 0.2 vs. 5.4 ± 0.3; and Total cholesterol (TC)=189 ± 4.0 vs. 162 ± 7.0; low density lipoprotein cholesterol (LDL-C), 111 ± 3.8 vs. 96 ± 5.0; triacyglycerol (TG), 202 ± 9.0 vs. 118 ± 5.3 mg/dL, respectively. The serum phosphorus level was significantly increased in CVD patients (mg/dL, patient vs. control, 5.1 ± 0.10 vs. 3.7 ± 0.1). Simple regression analyses revealed a highly significant positive correlation between serum phosphorus and TC, TG and FG. Thus the results demonstrate that the serum phosphorus level might be another parameter which is closely associated with diabetes and could also be used as a possible marker for the risk of CVD.

Syzygium cumini seed extract protects the liver against lipid peroxidation with concurrent amelioration of hepatic enzymes and lipid profile of alcoholic rats.

The in vitro oxidative stress induced by ethanol/Fenton's reaction in rat liver homogenates decreased significantly in the presence of Syzygium cumini seed extract, suggesting the protective effect of the seed extract against the oxidative stress in liver. To corroborate the in vitro effects by an in vivo experiment, 24 rats were divided into four groups: control, S. cumini seed-extract-administered (SE), 15% ethanol-fed (Alc) and Alc+SE rats. The oral administration of the extract (400 mg/kg BW.day) for 7 weeks significantly decreased the levels of liver LPO in the Alc+SE rats, suggesting that S. cumini seed not only obstructed the in vitro free radical production and subsequent oxidative stress, but also inhibited their in vivo formation. The oral administration of extract also reduced the enzyme activities of serum gammaglutamyl transferase, glutamate oxaloacetate transaminase and glutamate pyruvate transaminase and the levels of serum creatinine and blood urea nitrogen, serum/liver triglycerides and total cholesterol of the alcoholic rats. The levels of fecal cholesterol were increased by the extract. Fatty degenerations in liver and kidney were absent with S. cumini seed extract treatment. The results suggest that S. cumini seed may be a potential therapy for alcoholics and related dysfunctions by restraining oxidative stress.

Effects of Hilsa ilisa fish oil on the atherogenic lipid profile and glycaemic status of streptozotocin-treated type 1 diabetic rats.

1. The effects of oral administration of Hilsa (Hilsa ilisa) fish oil (1 g oil/kg bodyweight per day) on the lipid profile, platelet aggregation, anti-oxidative status and glycaemic control of streptozotocin (STZ; 90 mg/kg bodyweight)-treated type 1 diabetic rats were compared with those in fish oil-treated or untreated non-diabetic rats. 2. After 3 weeks of fish oil feeding, plasma total cholesterol decreased in both the non-diabetic and diabetic rats by 35 and approximately 10%, respectively, and triglyceride fell by 69 and 20%, respectively, compared with control rats. 3. Fish oil feeding decreased non-esterified fatty acids (NEFA) by 29% in diabetic rats but the NEFA level in non-diabetic rats was unaffected. 4. In non-diabetic and diabetic rats, platelet aggregation decreased by 49 and 37%, respectively, and total anti-oxidant status increased by 18 and 17%, respectively, after fish oil feeding. 5. Insulin levels increased by 27% in the fish oil-fed non-diabetic rats, whereas insulin levels were markedly decreased in diabetic rats. Glucose levels were not altered at all and fructosamine levels decreased by 29% only in fish oil-fed diabetic rats. 6. The results of the present study suggest that Hilsa ilisa fish oil may ameliorate the atherogenic lipid profile, platelet hyperaggregation and the anti-oxidative defence of STZ-diabetic rats and the amelioration is thought to be independent of the effects of Hilsa on glycaemic control.

Dietary mushroom (Pleurotus ostreatus) ameliorates atherogenic lipid in hypercholesterolaemic rats.

1. The effects of edible oyster mushroom Pleurotus ostreatus on plasma and liver lipid profiles and on the plasma total anti-oxidant status were estimated in hyper- and normocholesterolaemic Long Evans rats. 2. The feeding of 5% powder of the fruiting bodies of P. ostreatus mushrooms to hypercholesterolaemic rats reduced their plasma total cholesterol by approximately 28%, low-density lipoprotein-cholesterol by approximately 55%, triglyceride by approximately 34%, non-esterified fatty acid by approximately 30% and total liver cholesterol levels by > 34%, with a concurrent increase in plasma high-density lipoprotein-cholesterol concentration of > 21%. However, these effects were not observed in mushroom-fed normocholesterolaemic rats. 3. Mushroom feeding significantly increased plasma fatty acid unsaturation in both normo- and hypercholesterolaemic rats. 4. Plasma total anti-oxidant status, as estimated by the oxidation of 2,2'-azino-bis-[3-ethylbenz-thiazoline-6-sulphonic-acid], was significantly decreased in mushroom-fed hypercholesterolaemic rats, concomitant with a decrease in plasma total cholesterol. 5. The present study suggests that 5% P. ostreatus supplementation provides health benefits, at least partially, by acting on the atherogenic lipid profile in the hypercholesterolaemic condition.