Comparative study on lipid peroxidation and antioxidant vitamins E and C inFalciparum andVivax malaria

Reactive oxygen species are thought to be involved in the pathogenesis of Malaria. To assess the extent of oxidative stress, a study was conducted in patients withPlasmodium falciparum malaria andPlasmodium Vivax malaria. Plasma Thiobarbituric acid reactive substances (TBARS) were measured to assess the degree of lipid peroxidation. Antioxidant status was measured by estimating the levels of Vitamins E and C. Results were compared with age and sex matched control subjects. This study suggests that plasma TBARS levels were significantly increased in malaria patients. The patients withP. falciparum infection showed significantly increased levels of lipid peroxides when compared toP. vivax malaria. The antioxidant Vitamins E and C were decreased significantly in malaria patients in both the groups. Maximum decline in Vitamin C was observed inP. vivax malaria. Therefore it is been hypothesized that antioxidant Vitamins E and C could provide protection against the oxidative stress induced by malaria.     

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.     

Use of antigen prepared from in vitro culture ofPlasmodium falciparum for the detection of antibodies in sera from patients exposed toPlasmodium vivax

The blood stage parasites fromin vitro cultures ofPlasmodium falciparum were employed for the detection of malaria antibodies in patient sera havingP. vivax infections employing indirect fluorescent antibody (IFA) test. The antibody titres obtained for theP. vivax infected sera ranged from 1∶128 to 1∶1024. Control slides smeared with non-immune sera did not show any fluorescence. On the basis of the results of the present study it maybe concluded that antigen prepared fromin vitro cultures ofP. falciparum cross-reacts withP. vivax antibodies.     

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.     

Oxidants and antioxidants in hypoxic ischaemic encephalopathy

In the present study, the role of serum lipid peroxide and serum nitric oxide as oxidants and erythrocytic superoxide dismutase & serum vitamin E as antioxidants were determined in the 50 neonates with hypoxic ischaemic encephalopathy. (HIE) as against 25 healthy neonates as controls 50 patients of HIE were further divided into two groups i. e. mild and moderate HIE patients. All subjects were in the age group of 37–41 weeks of gestation. The levels of serum lipid peroxide, serum nitric oxide and erythrocytic superoxide dismutase were significantly elevated in both groups of neonates with HIE than those of controls (P<0.001), whereas serum vitamin E levels were significantly decreased in both groups of HIE patients than those of controls (P<0.001). A positive correlation was obtained between serum lipid peroxide and erythrocytic superoxide dismutase (r=+0.86). Alterations in the status of oxidants and antioxidants indicate role of free radicals in the development of HIE.     

Effect of vitamin E supplementation on oxidative stress in hemodialysis patients

Hemodialysis represents a chronic stress status for its recipients. Many hypotheses state that this may be associated with oxidative stress. Thus, there may be deficiency of antioxidants like erythrocytic superoxide dismutase, catalase, vitamin E or increased generation of free radicals like superoxide anions. A study was carried out to investigate oxidant and antioxidant status in chronic renal failure patients undergoing hemodialysis and effect of vitamin E supplementation on these two status. Blood samples were collected from patients before and after hemodialysis and from controls. The samples were analyzed for quantitation of MDA as index of lipid peroxide, nitric oxide, vitamin E, vitamin C and enzymatic antioxidants namely erythrocyte SOD and catalase. As compared to controls, the levels of serum MDA were significantly increased and activities of erythrocyte SOD and catalase, levels of serum nitric oxide, serum vitamin E and plasma vitamin C were significantly decreased both before and after hemodialysis. The efficiency of vitamin E therapy in hemodialysis patients was assessed by re-evaluating oxidant and antioxidant status of same patients after supplementation of vitamin E. Vitamin E supplementation caused decrease in serum MDA and increase in levels of serum nitric oxide, vitamin E, vitamin C and activities of erythrocytic SOD and catalase. Our results suggest the presence of oxidative stress and the possible preventive role of vitamin E therapy in hemodialysis patients.     

Evaluating role of oxidative stress in determining the pathogenesis of falciparum malaria induced acute renal failure

Serum malondialdehyde was measured in sixty-one falciparum malaria cases, which include thirty uncomplicated, and thirty-one complicated with acute renal failure. Twenty-six healthy individuals were also studied as controls. Serum malondialdehyde level was found to be significantly elevated in falciparum malaria induced acute renal failure cases when compared with uncomplicated falciparum malaria (p<0.001) and healthy controls (p<0.001). A positive correlation with the raised urea, creatinine and bilirubin levels were significant (r=0.62, p<0.025; r=0.65, p<0.05 and r=0.72, p<0.001 respectively) indicating the severity of complication with rise of lipid peroxides in falciparum malaria induced acute renal failure cases.     

Determination of immunoreactivity of<Emphasis Type="Italic">Plasmodium falciparum</Emphasis> antigens,serum dilutions and biomaterials

Immunoreactivity properties of serum dilutions andPlasmodium falciparum malaria antigens were measured and compared by ELISA technique using different ELISA plates to evaluate the role of antigens and serum dilutions for optimum binding. Also effort has been made to see the effect of reaction surface and material i.e. ELISA plates for binding capacity. Serological properties were estimated by ELISA methods for detection of malaria and determination of immunological characteristics. Three Pf antigens (PfAg) i.e. ring infected erythrocyte surface antigen: AR-1 (RESA), histidine-rich protein 2 antigen (HRP-2) and glycophospholipid antigen (grown and developed Pf antigen from PSJ-M strain): GPL1 have been used for serological testing of human blood samples by Enzyme Link Immunosorbant Assay (ELISA). 1∶100, 1∶1000 and 1∶10000 dilutions of Pf positive and negative serum (50 samples in each group) and 1∶1000 dilution of Pf antigens were used to measure immunoreactive properties by ELISA method. Result of PfAg-serum immunoreactivity study showed that GPL1 has the highest degree of immuno binding reactivity compared to other Pf antigens. HRP-2 and RESA antigens showed no significant difference to each other. Study also found that Costar and Fastec ELISA plates have a better Ag−Ab binding capability compared to immulon and Falcon plates at all dilutions of serum. Serum dilution of 1∶100 showed best binding and reactivity with Pf antigens followed by 1∶1000 and 1∶10000 showed lowest reactivity.     

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 (HbA_1c). 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), HbA_1c (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 HbA_1c (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.     

Oxidative stress in metabolic syndrome

As antioxidants play a protective role in the pathophysiology of diabetes and cardiovascular diseases, understanding the physiological status of antioxidant concentration among people at high risk for developing these conditions, such as Metabolic Syndrome, is of interest. In present study out of 187 first degree non-diabetic relatives and 192 non-diabetic spouses, 33.1% and 19.7% were found to have metabolic syndrome respectively. Subjects with metabolic syndrome (≥3 risk factors) had poor antioxidants status as reflected by significantly low levels of vitamin A, C & E and significantly increased (p<0.01) oxidative stress as compared to those without metabolic syndrome. At the same time serum insulin levels and insulin resistance were found to be significantly high (p<0.001) in metabolic syndrome. A strong positive correlation (r=0.946; p<0.001) between oxidative stress and insulin resistance was observed in metabolic syndrome. Low levels of antioxidants and increased oxidative stress with insulin resistance in metabolic syndrome suggests that besides therapeutic life style changes (TLC) as suggested in ATP III guidelines inclusion of antioxidant vitamins, fruits and vegetable could be beneficial to ward off the consequences of metabolic syndrome.     

Antioxidant status in neonatal jaundice before and after phototherapy

Phototherapy has been related to increased oxidative stress and lipid peroxidation. In the present study, thirty full term jaundiced neonates with appropriate weight were analyzed before and after completion of phototherapy for malondialdehyde (MDA), reduced glutathione (GSH), total thiols, vitamin C and superoxide dismutase (SOD) levels in hemolysate and albumin levels in plasma. These parameters were analyzed in cord blood samples of 20 healthy neonates as control. It was observed that levels of MDA were elevated significantly (p<0.001) in patients as compared to controls and that the levels increased significantly after phototherapy (p<0.001). Levels of SOD were also found to be increased significantly as compared to controls and the levels rose after phototherapy (p<0.001). On the other hand, the levels of non-enzymatic antioxidants such as GSH, total thiols and vitamin C were significantly low (p<0.001) as compared to controls and the levels decreased significantly after phototherapy (p<0.001). The plasma albumin levels also were found to be decreased significantly after phototherapy (p<0.01). Therefore, phototherapy increases oxidative stress and should be used with care.     

Oxidative stress and antioxidant status in cervical cancer patients

Cervical cancer (CaCx) is a global public health problem as it is the second most common cancer leading to the death of women worldwide. Many references revealed that the low levels of antioxidants induce the generation of free radicals leading to DNA damage and further mutations. In the present study attempt have been made to evaluate the levels of serum Lipid peroxide, Nitric Oxide (NO^.) Erythrocytic—Superoxide Dismutase (RBC-SOD), Vitamin-C, serum Copper (Cu) and serum Zinc (Zn). 120 patients were divided in 4 groups according to the increasing CaCx stages i.e. stage I, II, III & IV respectively. All the patients were around the age group of 25–65 years. 30 healthy women between the same age group were treated as controls. Highly significant increased values of MDA, NO^. and Cu were observed (p<0.001) whereas the activity of RBC-SOD, levels of Vitamin-C and Zn were significantly decreased in CaCx patients as compared with healthy controls (p<0.001). Cu/Zn ratio was found to be altered in CaCx patients. From our findings it can be concluded that the oxidative stress is induced among CaCx patients, which inturn increases the risk of CaCx.     

Effect of radiotherapy on serum sialic acid level in carcinoma cervix

Serum sialic acid levels were determined in 66 patients of carcinoma cervix and 25 control women matched for age. Mean sialic acid level was significantly elevated (p<0.001) in cancer cervix patients as compared to controls. Patients were treated with 50–65 Gy of radiation in 5–6.5 weeks. Post-therapy levels were again determined. Radiotherapy caused decline in serum sialic acid level in all the patients except one. Mean post-therapy level was significantly lower as compared to pretreatment level (p<0.001). It was also observed that fall in sialic acid level was significantly higher (p<0.05) in early stage patients as compared to late stage. No relationship was observed in the dose of radiation and fall in serum sialic acid level. A significantly higher fall in serum sialic acid level was observed in patients having complete clinical response as compared to partial or nonresponders to radiation treatment (p<0.001).     

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.     

Effect of supplementation of vitamin C and E on oxidative stress in osteoporosis

Osteoporosis encompasses a wide spectrum of conditions associated with imbalance of osteoclastic and osteoblastic activities. The increased activity of osteoclasts leads to increased free radical formation and hence lipid peroxidation. Present study probes into the role of antioxidants as a palliative treatment for osteoporosis. It involved 50 healthy controls and 75 clinically diagnosed osteoporosis patients. Both the groups underwent baseline assessment of biochemical markers viz. osteoblastic markers: serum Alkaline phosphatase. Free or ionic calcium and Inorganic phosphorus, osteoclastic markers: serum Tartarate resistant acid phosphatase and Malondialdehyde and the antioxidant status: serum Superoxide dismutase and Erythrocyte reduced glutathione. The osteoporotic group was then divided into groups A (Vitamin E-Evinal 400 mg), B (Vitamin C-Celin 500 mg), C (Vitamin E+C-Evinal+Celin) for antioxidant supplementation for a period of 90 days. The results reveal that there is significant fall in concentration of serum MDA (p<0.001), TrACP (p<0.01). Improvement in antioxidant status is reflected by significant rise in concentration of serum SOD (p<0.001) and erythrocyte GSH (p<0.001) after 90 days of antioxidant supplementation in osteoporosis. The findings indicate that on the whole bone status improved with prolonged antioxidant vitamin supplementation, which can be used as a palliative treatment for osteoporosis. The efficacy is not affected whether the vitamins are administered singly or conjointly.     

Plasma MDA and antioxidant vitamins in diabetic retinopathy

Hyperglycemia and dyslipidemia in diabetes mellitus induce increased lipid peroxidation and peroxyl radical formation, an important mechanism in genesis of microangiopathy. We took up a study on oxidative stress, measured by plasma MDA and antioxidant vitamin status in type − 2 DM patients with and without retinopathy and compared them with a control non-diabetic group. Lipid peroxidation marker MDA was significantly elevated (p < 0.001) in both the diabetic groups whereas, serum vitamin E and vitamin C registered a significant fall (p<0.001) as compared to controls. Our correlation study revealed a significant positive association between plasma MDA with both fasting and 2hr post prandial plasma glucose (r=0.81, p < 0.001, r=0.92, p <0.001) suggesting the role of hyperglycemia in free radical production. Plasma MDA also depicted significant positive relation (p< 0.001) with all lipid parameters except serum HDLc pointing the role of dyslipidemia towards lipid peroxidation. Plasma MDA level was also found to be negatively correlated with both the vitamins (p<0.001, p<0.001) in the study group explaining their protective consumption in the oxidative process prevailing in diabetic retinopathy.     

Oxidative stress and serum α<Subscript>1</Subscript> — Antitrypsin in smokers

The present study was undertaken to evaluate the levels of serum lipid peroxide, nitric oxide end poducts, erythrocytic superoxide dismutase activity and serum α ₁-antitrypsin in smokers. Total 90 active cigarette smokers were subdivided into Group I (subjects with smoking habit of less than 10 cigarettes per day) and Group II (with smoking habit of more than 10 cigarettes per day). In both groups lipid peroxide and nitric oxide end products were significantly increased with significantly decrease in erythrocytic superoxide dismutase activity and serum α ₁-antitrypsin as compared to controls. Our findings show enhanced oxidative stress and reduced α ₁-antitrypsin in cigarette smokers. Further increase in number of cigarettes per day exacerbates the oxidative stress with decrease in α ₁-antitrypsin.     

Glycosylated proteins and serum lipid profile in complicated and uncomplicated NIDDM patients

In the present study, 58 diabetic patients (type II) comprising uncomplicated (27) and complicated (31) cases along with 38 healthy matched controls were investigated for glycosylated hemoglobin (HbA₁), glycosylated serum proteins (GSP) and serum lipid profile viz. triacylglycerol, total cholesterol, HDL-cholesterol (HDL-C), VLDL-C, LDL-C and their ratios. HbA₁ and GSP were significantly elevated in diabetic group as a whole when compared with controls (p<0.01), but comparison of mean values of complicated and uncomplicated group revealed (though higher in complicated) that the difference between two groups was not statistically significant (p>0.05). Follow-up studies showed that while GSP levels reflect variations in blood glucose over a shorter period of 15 days, those of HbA₁ do so for about 3 months. HbA₁ correlated with fasting blood glucose (p<0.01), whereas GSP showed correlation with both fasting (p<0.01) and postprandial (p<0.05) blood glucose. Hypercholesterolemia and hypertriacylglycerolemia were observed (p<0.01) in both the groups. However no significant change was seen in HDL-C (p>0.05) even after accounting for sex in both the groups. Interestingly TC:HDL-C was elevated significantly (p<0.01) in the above groups. These findings seem to suggest that at the levels of hyperglycemia, hypercholesterolemia and hypertriacylglycerolemia observed, some patients are prone to develop complications earlier.     

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.     

Scientific evaluation of some Ayurvedic preparations for correction of iron deficiency and anemia

A number of preparations are available in Ayurved for treatment of anemia and iron deficiency. This study was designed to evaluate efficacy of some of them. Six most commonly used Ayurvedic iron containing preparations (Navayasa Curna, Punarnavadi Mandura, Dhatri Lauha, Pradarantaka Lauha, Sarva-Juara-Hara Lauha and Vrihat Yakrdari Lauha) were given in a dose of 250 mg b.d. for 30 days to six groups of iron deficient anemic patients; each group consisting of 20 patients. A control group was given Allopathic preparation—Irex-12, (containing—ferrous fumarate, vitamin C, folic acid and vitamin B₁₂); 1 capsule daily for 30 days for comparison. All hematological and iron parameters were determined before and after completion of treatment. The results showed that there was statistically significant rise (p<0.001) in all of them—Hb, PCV, TRBC, MCV, MCH, MCHC and plasma iron, percent saturation and plasma ferritin. Total iron binding capacity decreased significantly (p<0.001). The response of most of Ayurvedic preparations was better than Allopathic preparation and there was no side effect as observed with iron salts The Hb regeneration rate was 0.10 g/dl/day for Allopathic preparation; while it was above this value for all Ayurvedic preparations exceptPradarantaka Lauha which was least effective.Sarva-Juara-Hara Lauha was the drug of choice as Hb regeneration with it was highest 0.16 g/dl/day. Upon analysis of Ayurvedic drugs, these results were found to be consistent and correlated with iron content of the preparation.