Evaluation of oxidative stress and antioxidant status of pregnant women suffering from malaria in Cameroon

Oxidative stress is thought to be involved in the pathophysiology of malaria, especially in pregnancy where natural resistance is markedly reduced. In the present study we investigated oxidative stress in 315 pregnant women out of which 159 had Plasmodium falciparum malaria and 154 controls. We evaluated the level of lipid peroxidation products (MDA level) in the plasma, the activity of erythrocyte antioxidant defense enzymes, superoxide dismutase (SOD, EC: 1.15.1.1) and catalase (Cat, EC: 1.11.1.6) as well as the ability to resist oxidative stress by the FRAP (Ferric Reducing Ability of Plasma) assay. Total erythrocyte protein levels were also examined. For the two groups of patients, several differences between the biochemical parameters tested were found. Median parasitaemia in women with malaria was 25,392 parasites/μl of blood (Range1200–82000), while in controls we had no parasites found in thin and thick smears. Levels of lipid peroxidation products (MDA) were significantly higher in patients with parasitemia than in healthy asymptomatic volunteers (mean: 0.844 ± 0.290 and 0.384 ± 0.129 respectively, p<0.001). This MDA level was higher in primigravidea and also correlates well with parasite density (p<0.001). Catalase activity in erythrocytes of women with malaria did not differ statistically from that of controls. In contrast, SOD activity of patients with malaria was found to be significantly higher than that of controls (mean: 0.7899 ± 0.2777 and 0.4263 ± 0.2629 respectively, p<0.05). FRAP values declined, from parasitemic patients (1.4619 ± 0.6565) compare to controls (2.4396 ± 0.8883, p<0.05), particularly in the first and third trimester of gestation (p<0.05 and p<0.01 respectively). Finally, total erythrocyte protein concentrations of women with malaria did not differ from that of the controls. Our results suggest an imbalance between oxidants and antioxidants in pregnant women suffering from malaria, a situation which could lead to severe damage for either the mother or the fetus. Therefore, further research should be done to assess the potential benefits of antioxidant supplementation for the pregnant women suffering from malaria.     

Comparison between erythrocyte hemoglobin and spectrin glycosylation and role of oxidative stress in type-2 diabetes mellitus

New findings on organization of blood cell cytoskeleton represent an exciting aspect of modem cell biology and hematology, which is an interesting investigation to study diabetes. The present study was undertaken in 150 subjects. Out of these, 30 subjects were controls (Group I) and 30 were type-2 diabetics without any complication (Group II), while remaining 90 subjects were type-2 diabetics with complication (Group III). We determined erythrocyte spectrin and hemoglobin glycosylation and also estimated plasma lipid peroxide, nitric oxide and erythrocyte glutathione peroxidase activity to assess the status of oxidative stress. There was a significant increase in spectrin (P<0.001) and hemoglobin (P<0.001) glycosylation in Group II and III as compared to Group I and spectrin glycosylation was nearly three times more as compared to hemoglobin, whereas plasma levels of lipid peroxide (P<0.001) as well as nitric oxide (P<0.001) were found to be significantly increased and GPx activity (P<0.001) was significantly decreased in Group II and III as compared to Group I. However, it was also observed that spectrin (P>0.05) and hemoglobin (P>0.05) glycosylation was not significantly different in Group II and III. In contrast, there was significant rise in lipid peroxide (P<0.001), nitric oxide (P<0.001) and fall in GPx activity (P<0.001) in Group III when compared to Group II. Increased erythrocyte protein glycosylation and oxidative stress is clearly evident from our study. However, to understand the exact interplay between these two mechanisms, further studies are required.     

Depleted nitrite and enhanced oxidative stress in urolithiasis

Crystal aggregation and retention are critical events for the formation of kidney stones. There is a close association between crystal development and free radical activity in vivo. In the present study 30 subjects presenting with urolithiasis were included. Serum levels of total lipid peroxides, nitric oxide (as nitrite), α-tocopherol, plasma ascorbic acid (vitamin C) and erythrocyte superoxide dismutase activity were measured. These findings were compared with 30 age matched control subjects irrespective of sex. Student's ‘t’ test was applied for statistical analysis. There was a significant increase in lipid peroxides (p<0.001), where as significant decrease in nitrite (p<0.01) and α-tocopherol (p<0.001) levels were observed. Plasma ascorbate (p>0.05) and erythrocyte superoxide dismutase activity (p>0.05) was also found to be decreased but the difference was not statistically significant which suggests that oxidative stress is evident in urolithiasis with depletion in antioxidant status where as decrease in nitric oxide may be less abetting in disease condition.     

Nychthemeral rhythmicity of urinary copper levels in healthy Indians and renal stone formers

Nychthemeral rhythmicity of urinary copper (Cu) was studied in 40 clinically healthy volunteers and 35 renal stone formers. A marked diurnal rhythm was noticed in urinary Cu levels in healthy subjects with maximum excretion at 0000 to 0600 hr followed by a progressive fall during the rest of the period reaching minimum between 1200 to 1800 hr in males and 0600 to 1200 hr in female volunteers. Similarly, all stone patients exhibited a definite rhythm in urinary Cu with significant amplitude and acrophases at 2145 hr for males and 2141 hr for female stone formers. However, there was no difference in mean 24-hr Cu excretion in male and female stone formers, whereas a significant change was observed in case of healthy subjects. Urinary Cu was found to be significantly increased in renal stone formers in comparison to healthy controls of either sex attributing a significant role of this overlooked trace mineral in the crystallization and/or activation of either of the process of stone formation in the renal tubules.     

Acid Phosphatase as a Marker in Malaria

The serum acid phosphatase (ACP) activity and Hemoglobin (Hb) levels were measured in malaria patients and nonmalarial fever patients. The results were compared with normal healthy control subjects. ACP was significantly increased (P < 0.001) in all the malaria patients. ACP was significantly higher in Plasmodium falcifarum malaria and mixed malaria when compared to Plasmodium vivax malaria. Hb levels were significantly decreased in all the malaria patients which indicates that malaria parasite uses host erythrocyte Hb as a major nutrient source. There is negative correlation(r = −0.478) between ACP and Hb in malaria patients, which is highly significant. These results suggest that the measurement of ACP could be used as a marker for malaria.     

Increased erythrocyte resistance to osmotic lysis in the acute hepatitis caused by true hepatotropic viruses non-A, non-B (nanb)

An unexpected increase in erythrocyte osmotic resistance during viral hepatitis in two patients prompted study of effects of hepatitis (in Karad) on osmotic resistance. The test was performed by placing erythrocytes in saline solutions of decreasing osmolarity and osmotic fragility defined in terms of the saline concentration at which hemolysis begins. Study included 44 infected subjects (acute viral hepatitis non-A non-B). All forty four infected subjects showed abnormal results when compared to normal subjects. increased erythrocyte osmotic resistance i.e. hemolysis begins between 0.45% to 0.40% of NaCl and is completed between 0.25% to 0.20% NaCl, whereas in normal subjects. hemolysis begins between 0.50% to 0.45% NaCl and is completed between 0.35% to 0.30% NaCl. Osmotic resistance was observed with increase in total bilirubin (mean±SD) (4.6 ±3.6), direct bilirubin (3.0±2.5), SGOT (58.8±55.5) SGPT (114.2±150.3) activity. Physiologic shifts in erythrocyte osmotic resistance may be due to changes in membrane lipid ratio.     

Biochemical Effects of Lead Exposure on Systolic &; Diastolic Blood Pressure,Heme Biosynthesis and Hematological Parameters in Automobile Workers of North Karnataka (India)

The purpose of this study was to find out the effect of lead exposure on systolic and diastolic blood pressure, heme biosynthesis related and hematological parameters of automobile workers. For this study 30 automobile workers were selected and compared with 30 age matched healthy control subjects. Significantly increased blood lead (364%, P < 0.001) and urinary lead (176%, P < 0.001) levels were observed in automobile workers (study group) as compared to controls. Systolic blood pressure (5.32%, P < 0.05) and diastolic blood pressure (5.87%, P < 0.05) were significantly increased in the automobile workers as compared to controls. The significantly decreased non-activated erythrocyte δ-aminolevulinic acid dehydratase (δ-ALAD) (−18.51%, P < 0.01) and activated δ-ALAD (−13.29%, P < 0.05) levels were observed in automobile workers as compared to normal healthy control subjects. But the ratio of activated/non-activated δ-ALAD was significantly increased (43.83%, P < 0.001) in automobile workers as compared to controls. Excretions of δ-aminolevulinic acid (83.78%, P < 0.001) and porphobilinogen (37%, P < 0.001) in urine were significantly increased in the study group as compared to the controls. In automobile workers heamoglobin (−11.51%, P < 0.001), hematocrit (−4.06%, P < 0.05), mean corpuscle volume (−3.34%, P < 0.05), mean corpuscle hemoglobin (−5.66%, P < 0.01), mean corpuscle hemoglobin concentration (−7.67%, P < 0.001), red blood cell count (−14.6%, P < 0.001) were significantly decreased and total white blood cell count (11.44%, P < 0.05) increased as compared to the controls. The results of this study clearly indicate that the absorption of lead is more in automobile workers and it affects on blood pressure, heme biosynthesis and hematological parameters observed in this study group.     

Lipid peroxide levels and antioxidant status in alcoholic liver disease

The present study was conducted to evaluate some of the components of antioxidant defense system and oxidative damage in 20 male patients of alcoholic liver disease (ALD). The results were compared with 20 healthy male smokers and 20 healthy male non-smokers volunteers. Patients were subjected to detailed clinical examination and laboratory investigations. Blood samples were collected for estimating reduced glutathione (GSH), total thiols (T-SH) malondialdehyde (MDA), transaminases (AST, ALT), glutathione-S-transferease (GST) and gammaglutamyl transferase (GGT). Serum aspartate amino transferase (AST)/alanine amino transferase (ALT) ratio was significantly (p<0.01) reduced in ALD patients as compared to the controls. However, the core of utility of MDA and GST was found to be significantly (p<0.01) increased in ALD patients compared to controls. There was a significant negative correlation of MDA with both GSH and TSH. Plasma GGT levels were significantly (p<0.01) increased in alcoholics and the enzyme showed a significant positive correlation with MDA. These results give enough evidence of increased oxidative stress and compromised antioxidant defense system in patients with ALD.     

Somatic DNA Damages in Cardiovascular Autonomic Neuropathy

Cardiovascular autonomic neuropathy (CAN) is one of the most clinically significant complications of diabetes mellitus. Even though many ethological factors have been attributed for the pathogenesis of this disease no attempts were made to correlate DNA damage as a causative factor. Hence the present study was undertaken to asses the extent of somatic DNA damages by cytokinesis-block micronuclei assay (CBMN). An attempt is also being made to correlate the habits and/or risk factors and socioeconomic status with CAN. The CBMN frequency of 46 patients suffering from autonomic neuropathy was compared with that of 25 healthy age and sex matched controls. All the subjects were suffering from type 2 diabetes for at least 8 years and have varying degrees of coronary artery diseases. The mean CBMN frequency of the patients was statistically higher than that of the healthy control subjects (P < 0.05). The CBMN frequency was found to be significantly altered in CAN patients who where physical inactivity and smoking. A significant correlation could also be observed between CAN and smoking, diabetes mellitus, hypertension, dyslipidemia, abdominal obesity, and physical activity.     

Maternal and fetal indicators of oxidative stress during intrauterine growth retardation (IUGR)

The present study demonstrates the possibility of increased lipid peroxidation and protein oxidation in both maternal and fetal erythrocytes as markers of oxygen radical activity during intrauterine growth retardation. The erythrocyte MDA levels were significantly elevated in mothers of IUGR babies when compared to controls (p<0.01). The endogenous protein damage due to oxidative stress was significantly higher in IUGR mothers when compared to controls (p<0.05). Similarly the proteolytic activity in erythrocyte lysates against oxidatively damaged hemoglobin was significantly increased in mothers of IUGR babies compared to controls (p<0.001). In fetuses born with IUGR, both lipid peroxidation and proteolytic activity were significantly increased when compared to normal newborns (p<0.01). The result of this study indicates that oxidative stress was induced both in IUGR babies and their mothers which is manifested as increased lipid peroxidation and protein oxidant damage.     

Comparing age-wise reference intervals for serum creatinine concentration in a “Reality check” of the recommended cut-off

An increase in the communication within the healthcare services, both nationally and internationally, has strengthened the need for harmonization of measurements and reference intervals in laboratory medicine. In the present report, the calculated reference interval for serum creatinine (sCr) levels of healthy normal individuals (n=1121) in different sex and age groups are compared with the established interval. The calculated reference interval for sCr level was 0.4–1.3 mg/dL and 0.6 to 1.3 mg/dL in the age groups of 21–40 and 41–60 years respectively. The difference between the mean sCr values in total males and total females (age range 21–60 years) was statistically significant (p<0.0001); When male and female subjects were analyzed age-group wise, the data showed a significant difference in mean sCr values (p<0.0001) in three age groups (21–30, 31–40 and 41–50 years) however, in older age group (51–60 years), the difference was non-significant (p=0.07). The reference ranges were 0.7–1.3 and 0.4–1.0 mg/dL for males and females respectively where the lower limit was 0.1–0.2 units less than that of standard limits. An increase in the mean value of sCr was observed particularly in females with an increase in age. Hence it is of interest to validate an age specific reference ranges for sCr in our population.     

Serum gamma glutamyl transferase (γ-GT) activity in alcoholics with liver abscess and chronic alcoholics

Gamma Glutamyl Transferase (γ-GT) is a microsomal enzyme. Its activity in serum is assayed in alcoholics with liver abscess and alcoholics without liver abscess after screening by ultrasonography. The enzyme activity in serum is increased by 334% in the patients with liver abscess compared to normal control subjects (p<0.0001), and 172% when compared to patients with chronic alcoholism without liver abscess (p<0.0001). The enzyme activity was increased by 59.3% in the serum of patients without liver abscess compared to the normal control subjects (p<0.001). Measurement of γ-GT in serum appears to be a sensitive index in the diagnosis of alcoholics with liver abscess.     

Current Scenario of Prevalence of Vitamin D Deficiency in Ostensibly Healthy Indian Population: A Hospital Based Retrospective Study

25-hydroxy vitamin D [25(OH) vit D] deficiency is a serious public health problem, particularly in the Indian sub-continent. The objective of the present study was to study the prevalence of 25(OH) vit D in different age groups. The data of 25(OH) vit D assay of 26,346 ostensibly healthy individuals, enrolled under executive health checkup at Medanta The Medicity, Gurgaon, over a period of 3 years, were extracted from the hospital information system and reviewed extensively. 25(OH) vit D deficiency (VDD) was defined as 25(OH) vit D < 20 ng/ml, insufficiency (VDI) as 25(OH) vit D between 20 and 40 ng/ml and 25(OH) vit D sufficiency (VDS) as 25(OH) D > 40 ng/mL. 25(OH) vit D deficiency (VDD + VDI) was observed in 93 % of the subject population. Maximum number of the subjects belonged to the age group of 41–60 years. 59 % had frank 25(OH) vit D deficiency when cut off level was <20 ng/mL. Mean value of 25(OH) vit D in our subjects was 21.4 ± 14.4 ng/mL. Significant difference in 25(OH) vit D level was observed in between male and female subjects. Simultaneously 25(OH) vit D levels were significantly lower in the patient visited hospital in winter-spring season than the summer-autumn season (p > 0.001). Our study demonstrates a high prevalence of 25(OH) vit D deficiency in an ostensibly healthy Indian population. There is a need for redefining our reference ranges according to our population and extensively improving the status of vitamin D.     

Anomalous Activity Measurements of Creatine (Phospho) Kinase,CK-MB Isoenzyme in Indian Patients in the Diagnosis of Acute Coronary Syndrome

In the present study, the cause of suspected false-positive (anomalous) values for CK-MB activity, in Indian patients investigated for ACS. Total serum CK and CK-MB activity, serum Troponin I were measured and CK-MB as a percentage of the total CK activity (%CK-MB) calculated. CK-MB was also estimated using densitometry and CK-MB mass assay. Anomalous specimens were tested for the presence of CK isoenzymes. In 22 healthy subjects, 11 male and female, the %CK-MB ranged from 3.6 to 30.2. In 11 male patients, with proven ACS, the %CK-MB was from 4.0 to 17.5. The cut off for anomalous CK-MB activity values was set as >33.0%. In 35 patients with anomalies, total CK values ranged from 39 to 231 U/L, CK-MB from 30 to 161 U/L. Investigation of CK isoenzymes, showed 10 patients had a CK-BB band, 14 an intermediate band between CK-MM and CK-MB (macro-CK type 1), 7 had a cathodal band (macro-CK type 2), and 3 had a band intermediate between CK-MB and CK-BB. This later band does not seem to have been previously reported. Against the CK-MB mass assay, the activity assay showed no correlation, in 43 patients (19 M, 24 F), Pearson coefficient (R²) was 0.006. The CK-MB immunoinhibition assay is better described as measuring “non-CK-MM activity.” A %CK-MB activity >6% as a marker of ACS is not valid in our patient population. Laboratories should not use only CK-MB activity as a biochemical marker of ACS.     

hs-CRP and oxidative stress in young CAD patients: A pilot study

Premature coronary artery disease (CAD) is common in India. We, therefore, studied oxidative stress, dyslipidemia, and high sensitivity-C reactive protein (hs-CRP) levels in young CAD patients. Present study consisted of male CAD patients below 40 years and age and sex matched healthy controls (n=30 each). Fasting blood samples were analyzed for serum lipid profile, malondialdehyde, antioxidant enzymes and hs-CRP levels. Dyslipidemia was observed in 90% of the young CAD patients, of which 72.2% showed increased serum triglycerides and decreased HDL-cholesterol. LDL-cholesterol levels were high in 77.8%. Serum malondialdehyde and hs-CRP levels were increased significantly (p<0.0001) as compared to controls. hs-CRP levels were in high risk range in all the young patients. However, glutathione peroxidase activity was reduced significantly (p<0.05). Our data suggests that elevated hs-CRP levels along with dyslipidemia and oxidative stress adds to the predictive value of premature CAD in young Indians.     

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.     

Effect of Interaction Between Duration of Alcohol Consumption and Alcohol Dependence on Thyroid Function Test: Cross Sectional Observational Study

Dependence on alcohol, nicotine and duration of alcohol consumption are known to alter thyroid function tests. This study was conducted to assess the effect of interaction between the duration of alcohol consumption and alcohol dependence on TFT. The subjects consisted of 38 male patients with alcohol dependent syndrome co morbid with nicotine dependent syndrome, 33 male patients with alcohol dependent syndrome and 30 male normal healthy volunteers. Liver function tests, haematological parameters and thyroid function tests were assayed. Two way multivariate ANOVA was used to assess the interaction effect by SPSS 21 package. Multivariate analysis of combined TFT levels revealed no significant (P = .078) difference amongst groups based on alcohol dependence, significant difference (P = .001) amongst groups based on duration of alcohol consumption and no significant (P = .604) interaction effect between duration of alcohol consumption and alcohol dependence. Tests of between subject effects for individual TFT revealed significant (P = .014) difference in T3 between groups based on alcohol dependence, significant difference in the levels of fT4 (P = .001), T3 (P = .07) and T4 (P < .001) between groups based on duration of alcohol consumption was observed. Interaction between the effect of duration of alcohol consumption and alcohol dependence for individual TFT did not reveal any significance. fT4, TSH and T4 levels were significantly low in persons consuming alcohol for more than 20 years. TSH levels were significantly low in ADS compared to controls. Significant decrease in the levels of thyroid hormones was observed as the duration of alcohol consumption increased.     

Prostate specific antigen in patients of benign prostate hypertrophy and carcinoma prostate

Prostate Specific Antigen (PSA) has emerged as the most applicable and important tumor marker for carcinoma prostate. In the present study PSA was determined in serum of healthy subjects, patients of benign prostate hypertrophy (BPH) and Carcinoma Prostate (Ca−P) to evaluate its diagnostic efficiency in day to day management of prostate cancer patients and in differentiating patients of early prostate cancer from those with BPH. Receiver operating characteristic curve (ROC) revealed 2 ng/ml and 10 ng/ml cut off serum PSA level for BPH and untreated carcinoma prostate patients (Ca−P). An extremely significant increase (P<0.0001) was observed in mean PSA concentration in BPH patients and adenocarcinoma prostate patients when compared to healthy males. Clinical relevance of PSA was highlighted by a case study of cancer patient prior to any therapy till death.     

Studies on biochemical changes with special reference to oxidant and antioxidants in malaria patients

Oxidative stress plays an important role in the development of malarial anemia. The present study was undertaken to study the role of oxidant and antioxidants in the patients ofPlasmodium falciparum malaria (n=25),Plasmodium vivax malaria (n=25) as against the normal control subjects (n=25). The parameters included are the hematological [hemoglobin, erythrocyte adenosine deaminase (ADA) activity, ADP-induced platelet aggregation] and serum total lipid peroxide as an index of oxidative stress and antioxidants [erythrocytic superoxide dismutase (SOD) activity, serum vitamin E] & serum iron. Significant alterations in all above parameters were noted in both groups of malaria patients as compared to control subjects. Maximum significant alterations in hematological parameters were noticed inP. falciparum infection as compared toP. vivax malaria (p<0.001). Substantial rise in serum total lipid peroxides and a significant reduction in antioxidants such as serum vitamin E and serum iron were noted inP. falciparum malaria as compared toP. vivax malaria (p<0.001), whereas maximum decline in erythrocytic SOD activity was observed inP. vivax infection as compared toP. falciparum malaria (p<0.05). Follow-up examination revealed the restoration of the levels of all biochemical parameters to the normal level after 20 days of antimalarial therapy. The study specified severity ofP. falciparum malaria and also functional duality of oxidant.     

Biochemical markers for alcohol consumption

A variety of laboratory tests are available to assist in the diagnosis of alcohol consumption and related disorders. The levels of intake at which laboratory results become abnormal vary from person to person. Laboratory tests are particularly useful in settings where cooperativeness is suspected or when a history is not available. Several biochemical and hematological tests, such as γ-glutamyltransferase (GGT) activity, aspartate aminotransferase (AST) activity, high-density lipoprotein cholesterol (HDL-C) content of serum, and erythrocyte mean corpuscular volume (MCV) are established markers of alcohol intake. Their validity as markers is based largely on correlations with recent intake at a single time point and on decreases in elevated values when heavy drinkers abstain from alcohol. These readily available laboratory tests provide important prognostic information and should be integral part of the assessment of persons with hazardous alcohol consumption. There are several other markers with considerable potential for more accurate reflection of recent alcohol intake. These include carbohydrate deficient transferrin, β-hexosaminidase, acetaldehyde adducts and the urinary ratio of serotonin metabolites, 5-hydroxytryptophol and 5-hydroxyindoleacetic acid. These markers provide hope for more sensitive and specific aids to diagnosis and improved monitoring for intake.