Mungli Prakash

Relationship between free iron and glycated hemoglobin in uncontrolled type 2 diabetes patients associated with complications.

Free iron in serum has been found in several disease conditions including diabetes. In the present work, we studied the relationship between free iron, fasting blood glucose (FBG) and glycated haemoglobin (HbA1c). Study was carried out on 50 type 2 diabetes cases under poor glycemic control associated with complications, 53 type 2 diabetes cases under good glycemic control and 40 healthy controls. We estimated free iron, both ferrous (Fe+2) and ferric (Fe+3) form, protein thiols, lipid hydroperoxides, FBG, HbA1c and serum ferritin levels in serum. There was a significant increase in free iron in Fe+3 state (p <0.01), HbA1c (p<0.01), serum ferritin (p<0.01), lipid hydroperoxides (p<0.01) and significant decrease in protein thiols (<0.01) in diabetes cases under poor glycemic control compared to diabetes cases under good glycemic control and healthy controls. Free iron correlated positively with HbA1c (p<0.01). Poor glycemic control and increase in glycation of haemoglobin is contributing to the increase in free iron pool which is known to increase oxidant generation.

Paraoxonase: Its antiatherogenic role in chronic renal failure

Paraoxonase (PON) is an aryldialkylphosphatase, which reversibly binds and hydrolyzes organophosphates. The PON family has three members (PON1, PON2 and PON3); they share structural properties and enzymatic activities. PON1 is shown to reside over high density lipoprotein (HDL) and has both antioxidant and antiatherogenic functions. Function of PON2 and PON3 are speculative and still under research. Several methodologies were developed over the years to determine the activity and mass of PON1, of which spectrophotometer-based methods using certain chemicals as substrate predominate. Several studies have shown decreased levels of PON1 in chronic renal failure (CRF) patients, particularly those on hemodialysis. The role of PON1 in development of cardiovascular disease has drawn considerable attention in recent years. Several authors have shown decreased levels of HDL and PON1 activity in CRF patients on hemodialysis and reported this to be a risk factor in the development of CVD. Enhancement or maintenance of the PON1 activity may prevent development of CVDs and its consequences in patients on hemodialysis.

Copper and ceruloplasmin levels in relation to total thiols and GST in type 2 diabetes mellitus patients

Presence of oxidative stress in type 2 diabetes mellitus (DM) is well proved. Current study was undertaken to know the relation between fasting plasma glucose (FPG) and copper along with antioxidants like total thiols and ceruloplasmin, and antioxidant enzyme glutathione S transferase (GST). The study group consisted of a total of 201 subjects which included nondiabetic healthy control subjects (n = 78) and diabetic patients (n = 123). Plasma total thiols, GST, copper and ceruloplasmin levels were measured all the subjects using spectrophotometric methods and FPG levels were determined in clinical chemistry analyzer Hitachi 912. There was significant increase in FPG (P<0.001) and copper (P<0.001) and decrease in ceruloplasmin (P<0.001) and protein thiols (P<0.001) in type 2 DM cases compared to healthy controls. There was no significant change in GST between type 2 DM cases and controls. There was significant negative correlation of FPG with antioxidants like ceruloplasmin (r = −0.420, P<0.001) and total thiols (r = −0.565, P<0.001). Protein thiols correlated positively with ceruloplasmin (r = 0.364, P<0.001). Our study indicates possible increase in copper mediated generation of ROS leading to increased consumption of available antioxidants in the body.

Serum Paraoxonase 1 Activity Status in Patients with Liver Disorders

Background/Aim: Paraoxonase 1 (PON1) is an esterase, exclusively synthesized by liver. The present study has two objectives: to determine the PON1 activity status in various disorders associated with hepatocellular damage and to correlate the changes of PON1 activity with the standard liver function and fasting lipid profile tests in these disorders. Patients and Methods: The study groups consisted of 95 patients with liver diseases including acute viral hepatitis (14), cirrhosis with portal hypertension (33), leptospirosis (14), sepsis and multi organ failure (15), left ventricular failure (9), and falciparum malaria (10); and 53 healthy controls. Serum PON1 activity was measured manually using spectrophotometer. Liver function test parameters and fasting lipid profile were performed in clinical chemistry auto analyzer (Hitachi 912). Results: The serum PON1 activity in patients with acute viral hepatitis and sepsis decreased significantly (P<0.001) and moderately in falciparum malaria (P<0.05). However, in patients with cirrhosis, leptospirosis and left ventricular patients, its activity did not change significantly. On applying Pearson correlation, serum PON1 activity correlated positively with high-density lipoprotein-cholesterol (HDL-C) in patients with sepsis (r=0.633, P<0.05), left ventricular failure patients (r=0.814, P<0.05) and negatively with acute viral hepatitis patients (r=– 0.528, P<0.05). Conclusion: PON1 activity has decreased significantly in acute viral hepatitis, sepsis with multi organ failure and falciparum malaria patients. Determination of PON1 activity may serve as a useful additional test in assessing these conditions.

Determination of oxidative stress markers and their importance in early diagnosis of uremia-related complications

The existence of oxidative stress and the higher incidence of cardiovascular diseases in association with uremia is well proved. The uremic status of serum copper, ceruloplasmin (CP), protein thiols, malonyldialdehyde (MDA), and glutathione S-transferase (GST) levels was studied. The study was carried out on 51 chronic renal failure (CRF) patients who were not on hemodialysis therapy and on 42 healthy controls. Serum urea, creatinine, and MDA levels were found to be significantly increased (P < 0.001), and total protein, albumin, protein thiols, and copper levels were found to be significantly decreased in CRF patients compared to normal controls (P < 0.001). Ceruloplasmin levels were decreased significantly (P < 0.05), and there was no significant change in serum GST levels in CRF patients compared to normal controls. In conclusion, the significant increase in levels of MDA, and the decrease in levels of protein thiols, CP, and copper in uremia patients when compared to controls, reconfirms the presence of stress in this patient population. In view of the changes in other markers of oxidative stress, this absence of any significant change in the activity of GST in uremia patients compared to controls, warrants further study.

Urinary Protein Thiols In Different Grades of Proteinuria

Total thiol status of plasma, especially thiol groups over protein contributes maximum to the plasma antioxidant status of the body. Serum protein thiols were found to be decreased in various disease conditions including chronic renal failure patients. Only few studies determined the levels of urinary protein thiols in disease conditions. The current study was designed to know the levels of urinary protein thiols in patients with different grades of proteinuria. The study was conducted on urine of 40 healthy controls and 61 cases with proteinuria. Based on proteinuria cases were further divided into two groups; group I - microproteinuria (150–300 mg protein/d), 32 cases, group II - frank proteinuria (>300 mg protein/d), 29 cases. Urinary thiol levels were determined by spectrophotometric method using dithionitrobenzoic acid. A significant decrease (p<0.01) in urinary thiol in group I and group II cases was observed in present study and this decrease was associated with proteinuria.

Determination of urinary peptides in patients with proteinuria

Although considered useful in the diagnosis and prognosis of renal diseases, proteinuria can only be detected after significant renal paranchymal changes. There is considerable interest in the estimation of urinary peptides as an early marker of renal disease. In the current study, we have estimated urinary peptides in patients with different grades of proteinuria. Twenty-four hour urine samples were collected from 138 subjects and classified into three groups based on the urine protein excreted: group I (normoproteinuria, 0–150 mg/day, n = 37), group II (microproteinuria, 150–300 mg/day, n = 31), and group III (macroproteinuria, > 300 mg/day, n = 70). Urine proteins were determined using Bradfords method and urinary peptide levels were determined by subtracting Bradfords value from the Lowry value of the same sample. There was a significant decrease in the levels of urinary peptides in group III compared to group I (P < 0.01), however, there was no difference in peptides between groups I and II. The percentage of urinary peptides was decreased in both groups II and III compared to group I (P < 0.01), and there was a significant difference in % urinary peptide content in group II compared to group III (P < 0.01). On correlation, % urinary peptides correlated negatively with urinary proteins/g creatinine (r = - 0.782, P < 0.01) and positively with urinary peptides/g creatinine (r = 0.238, P < 0.01). Our data suggest that there is a marked decrease in urinary peptide levels with an increase in proteinuria. This may suggest impaired tubular protein reabsorption and degradation capacity of renal tubules.

Transitional Metal Ions Induced Damage to Biomolecules: Role of Ferryl and Perferryl Radicals

Objective: To investigate the effect of oxidized transitional metal (ferric and cupric) ions on the amino acids. Methods: 25 mmol/L hydroxyproline and 25 mmol/L histidine were incubated with 500μL Fe＞ and Cu(superscript 2+) ions at pH 7.4 and 37℃ for 30 mins in separate test tubes. Then 500 μL of 1% thiobarbituricacid (TBA) was added to the incubated amino acids followed by addition of 500μL of glacial acetic acid. The resultant mixture was vortexed and heated at 100℃ for 30 min. Absorbance readings were noted after cooling to room temperature. The experiment was repeated in the presence of various reagents, like hydroxyl radical scavengers, antioxidant enzymes, and reducing agents and metal ion chelators. Results: The pink chromogen formed with the absorbance maxima at 524 ran, AND shifted to 560 nm in alkaline pH. The absorbance was expressed as TBA-adduct in MDA units. The TBA-adduct decreased in the presence of reducing agents and metal ion chelators. Antioxidant enzymes and hydroxyl radical scavengers did not show any effect. Conclusion: Transitional metal ions in their oxidized state showed significant damage to amino acids, hydroxyproline and histidine. The results indicate the possible rule played by high-valent oxo-iron species, ferryl and perferry radicals in damaging biomolecules.