Pragna Rao

Reevaluation of the phenol-sulfuric acid reaction for the estimation of hexoses and pentoses

Evidence is provided to show that in the conventional phenol-sulfuric acid reaction procedure, phenol underwent sulfonation in situ and the phenolsulfonic acid formed decreased the color intensity for hydroxymethyl furfural (HMF), furfural, and many hexoses and pentoses tested. A modified method is described to overcome this problem in which phenol was added after the dehydration of carbohydrates by sulfuric acid and after cooling the system. The color intensity around 475-485 nm for different compounds was fairly proportional to the amount of furfural derivatives (absorption at 310-320 nm) formed from the sugars in the modified method unlike in the conventional procedure. The studies also show that for condensation of HMF derivatives with phenol, heat is not necessary. The color intensity in the modified method also increased compared to that in the conventional method. The increase in the modified method compared to that in the conventional method was 6.0-fold for furfural, 9.1-fold for hydroxymethyl furfural, 3.7-fold for fructose, 2.3-fold for xylose, and 2.0-fold for glucose and arabinose. The possible reasons for this differential increase are discussed.

Erythrocyte indicators of oxidative stress in gestational diabetes

Foetuses born to mothers with gestational diabetes are at increased risk of developing respiratory distress, foetal macrosomia, foetal anomalies and platelet hyperaggregability. High blood glucose level induces oxidative stress and decreases antioxidant defences. The present study discusses the possibility of lipid peroxidation and protein oxidation in both maternal and foetal erythrocytes as an indicator of oxygen radical activity. The level of lipid peroxidation and protein oxidation in erythrocytes was estimated in 20 mothers with gestational diabetes and their newborns. The maternal age varied between 19 and 42 y and foetal age ranged between 34 and 39 weeks. The proteolytic activities in the erythrocyte lysates obtained from mothers with gestational diabetes and their newborns were significantly greater [(mean ± SD) 24.41 ± 9.05 and 16.70 ± 3.36μM of amino groups/g haemoglobin, n= 20, respectively] than those from control group (10.18 ± 4.84 and 14.64 ± 6.21 μM amino groups/g haemoglobin, n= 15, respectively; p < 0:05 in both cases). Similarly erythrocyte malondialdehyde levels were significantly elevated in babies born to mothers with gestational diabetes (10.11 ±2.21 nM/g haemoglobin) when compared to controls (6.8 ± 3.75 nM/g haemoglobin) (p < 0:05). In the erythrocytes of mothers with gestational diabetes, malondialdehyde levels correlated significantly with glycated haemoglobin levels (p < 0:01). The results of this study indicate that the oxidative stress induced by gestational diabetes manifests as increased lipid peroxidation and protein oxidative damage in the erythrocytes of both mothers with gestational diabetes and their newborn infants.

Further studies on the mechanism of phenol-sulfuric acid reaction with furaldehyde derivatives

Even though the chromogens formed from mannose and galactose showed comparable absorbances at 480 nm in the conventional (developer present during heat of dilution) and modified (developer reacted at room temperature after cooling; epsilon mannose = 13,700, galactose = 14,000) phenol-sulfuric acid reactions, shoulders in the region 420-430 nm were prominent in the former method. Fucose was 10 times less reactive in the modified method (epsilon = 800) than in the conventional method. 2-Formyl-5-furan sulfonic acid reacted equally efficiently in the two methods (epsilon = 40,800). 5-Methyl-2-furaldehyde, unlike the sulfonate derivative or 5-hydroxymethyl-2-furaldehyde, required heat for condensation with phenol. 2-Furaldehyde dimethylhydrazone reacted 25 times better to form a chromogen (epsilon = 40,500) in the modified phenol-sulfuric acid method. The possible roles of intermediates between hexoses and furaldehydes in forming chromogens and the effect of substitution at the 2- and 5-positions of furaldehyde on the rates of condensation with phenol for the observed differences between the conventional and the modified methods are discussed.

Maternal and fetal indicators of oxidative stress in various obstetric complications.

The present study demonstrates the incidence of increased lipid peroxidation and protein oxidation in both maternal and fetal erythrocytes as markers of oxygen radical activity in different complications of pregnancy. In fetuses born after premature rupture of membranes, lipid peroxidation was significantly elevated as indicated by increased malondialdehyde levels (p<0.05) as compared to controls. Proteolytic activity in the erythrocytes of mothers in this group was also significantly high (p<0.01). In patients delivered by lower segment cesarian section, lipid peroxidation and proteolytic activity in maternal erythrocytes were significantly high (p<0.05 and p<0.001 respectively). In patients with prolonged second stage of labour, lipid peroxidation and proteolytic activity in maternal erythrocytes was significantly higher than in controls (p<0.001 and p<0.05 respectively). In this group, endogenous protein damage due to oxidative stress was significantly high both in the mother and the fetus (p<0.001 and p<0.05 respectively).

Angiotensin-converting enzyme gene insertion/deletion polymorphism studies in Asian Indian pregnant women biochemically identifies gestational diabetes mellitus:

Introduction: Gestational diabetes mellitus (GDM) is defined as glucose intolerance first recognized during pregnancy. Insertion/deletion (I/D) polymorphism of a 287 bp Alu repetitive sequence in intron 16 of the angiotensin-converting enzyme (ACE) gene has been widely investigated in Asian Indian populations with different ethnic origins. The present study examined possible association between I/D polymorphism of the ACE gene and GDM in Asian Indian pregnant women. Methods: A total of 200 pregnant women (100 GDM and 100 non-GDM) were recruited in this study and I/D polymorphism of a 287 bp Alu1 element inside intron 16 of the ACE gene was examined by polymerase chain reaction (PCR)-based gel electrophoresis. Result: The distribution of the variants like II, ID, and DD genotypes of ACE gene showed differences between normal GDM versus non-GDM subjects, and the frequency of the ID+ DD Vs II genotype was significant (p=0.0002) in the GDM group. Conclusion: ACE gene polymorphism was associated with GDM in Asian Indian pregnant women.

SERUM LIPIDS AND MALONDIALDEHYDE LEVELS IN PRIMIPAROUS PATIENTS WITH PREGNANCY INDUCED HYPERTENSION

BackgroundPregnancy induced hypertension (PIH) contributes to 15.6% maternal mortality in India. In Behrampur, Orissa, maternal deaths due to PIH was 32%, which is twice the national incidence. Hence in this population, some factors associated with severity of PIH were studied. Serum lipid concentrations and malondialdehyde (MDA) levels were correlated with severity of PIH and birth weight of the neonate.Patients & Methods70 primiparous PIH patients and 20 healthy controls were studied. Serum lipids and MDA were estimated. Maternal blood pressures and birth weights of the neonate were recorded.Results and ConclusionSerum cholesterol, triglyceride, LDL, VLDL and MDA were significantly elevated in primiparous PIH patients when compared to control subjects. Average birth weight of babies born to mothers with PIH was less than those born to control subjects. The most significant factor was that in this geographical area, 26% of the primiparous patients with PIH were below 20 years of age while only 15% of the controls were less than 20 years, indicating that younger maternal age was a contributing factor to PIH.

Investigation of Calpain 10 (rs2975760) gene polymorphism in Asian Indians with Gestational Diabetes Mellitus

Background Type 2 Diabetes Mellitus (T2DM) and Gestational Diabetes Mellitus (GDM) are part of a heterogeneous and complex metabolic group of disorders that share common pathophysiological circumstances, including β-cell dysfunction and insulin resistance. The protein Calpain 10 (CAPN10) plays a role in glucose metabolism, pancreatic β-cell insulin secretion, and thermogenesis. Objective Polymerase Chain Reaction–Restriction Fragment Length Polymorphism (PCR–RFLP) based genotyping of CAPN10 (rs2975760) polymorphism was carried out in T2DM and GDM with suitable controls for each of the pathologies from the same population. Genomic DNA was isolated from 787 participants, including 250 cases of T2DM, 287 pregnant women, of which 137 were identified as having GDM and the remaining 150 were confirmed as non-GDM, and 250 healthy control volunteers, and association analysis was carried out for genotypes and alleles. Results In the present study, T2DM was compared with healthy controls and was not found to be associated with the CAPN10 C allele (odds ratio, OR: 1.09; 95% CI = 0.8011–1.484; p = 0.5821). GDM also did not show any association when compared with non-GDM (OR: 1.124; 95% CI = 0.7585–1.667; p = 0.5606) respectively. Conclusion Our study suggests that the CAPN10 (rs2975760) polymorphism scrutinized in this study is not associated with T2DM and GDM.

MALNUTRITION- INFLAMMATION- ATHEROSCLEROSIS SYNDROME IN CHRONIC KIDNEY DISEASE

Chronic kidney disease is becoming a major health problem globally and in India an alarming number of about 8 million people are suffering from this disease. Patients undergoing hemodialysis have a high prevalence of protein-energy malnutrition and inflammation. As these two conditions often occur concomitantly in hemodialysis patients, they have been referred together as ‘malnutrition-inflammation-atherosclerosis syndrome’ to emphasize the important association with atherosclerotic cardiovascular disease. The three factors related to the pathophysiology in these patients are dialysis related nutrient loss, increased protein catabolism and hypoalbuminemia. Inflammation in Chronic Kidney disease is the most important factor in the genesis of several complications in renal disease. Pro-inflammatory cytokines like IL-1 and TNF-alpha play a major role in the onset of metabolic alterations in Chronic Kidney disease patients. Atherosclerosis is a very frequent complication in uremia due to the coexistence of hypertension, hyperhomocysteinemia, inflammation, malnutrition and increased oxidative stress, generation of advanced glycation end products, advanced oxidation protein products, hyperlipidemia and altered structural and functional ability of HDL. LDL-cholesterol, apolipoprotein (A), apolipoprotein (B), and Lp(a) are also associated with atherosclerosis. Studies have now provided enormous data to enable the evaluation of the severity of malnutrition-inflammation-atherosclerosis syndrome as well as effective monitoring of these patients.

Mitochondrial insertion-deletion polymorphism: role in disease pathology.

AIM Mitochondrial DNA (mtDNA) sequence variations are associated with a number of human diseases. The 9-bp repeat sequence, CCCCCTCTA, in the intergenic region of MTCO2 and MTTK genes of the mtDNA has been extensively used in phylogenic studies. The sequence has been reported to be polymorphic in south-east Asians in isolated cases of mt diseases. This is the first systemic study identifying the role of insertion-deletion polymorphism in human disease. RESULTS A total of 241 patients including those with cardiomyopathy, ataxias, and idiopathic neurological disorders along with 100 controls were screened; 2.9% of patients showed a single repeat (deletion) and 4.14% had three repeats (insertion), whereas all the controls had two repeats (normal). CONCLUSION This indicates that the 9-bp insertion-deletion repeat polymorphism plays a role in disease pathology, affecting the expression of the downstream genes of mtDNA and altering ATP generation.

Role of Glutamine Deamidation in Neurodegenerative Diseases Associated With Triplet Repeat Expansions

The pathological expansion of unstable trinucleotide repeats is known to cause neurodegenerative diseases. Trinucleotide repeat expansions might prove to be pathological through a variety of mechanisms, including alteration of DNA structure, transcription, RNA-protein interaction, and altered protein conformations/inter-actions. Deamidation of human proteins have been shown to regulate some time-dependent biological processes such as development and aging. In this paper we hypothesize the possible role of glutamine deamidation as a signaling event in the pathogenesis of neurodegenerative diseases associated with triplet repeat expansion.