Hyperammonemia and hepatic status during valproate therapy

The present study was conducted to assess correlation of ammonia levels with valproate levels in epileptic patients presenting with valproate toxicity and also whether liver enzymes and ammonia levels could serve as biochemical marker of valproate toxicity. 100 patients with epilepsy who had received valproate therapy for more than 12 months and had presented with valproate toxicity and 100 controls were included in the study. The serum valproate, ammonia and liver enzymes were measured in these subjects. In patients with valproate toxicity, the mean level of serum valproate was 110.91 ± 28.68 mg/dL (therapeutic range 50–100 mg/dL). Serum ammonia was higher (86.37 ± 39.90 μg/dL) in patients with valproate toxicity compared to controls (68.73 ± 30.07 μg/dL). Out of 100 patients, only 37 patients had serum valproate level > 120 mg/dL and 22 patients had raised levels of valproate as well as ammonia. Age < 30 years and serum ammonia > 69 μg/dL is risk factors for valproate toxicity. Serum ammonia, liver enzymes should be regularly investigated in patients on valproate therapy for early diagnosis of valproate toxicity.     

Adenosine deaminase and C-reactive protein in cerebrospinal fluid for differential diagnosis of tubercular meningitis in children

Adenosine deaminase and C-reactive protein in CSF were assessed in 72 cases of Tubercular meningitis, 24 cases of partially treated pyogenic meningitis, 20 cases of Aseptic meningitis and 8 cases of febrile seizures. Mean Adenosine deaminase value was 12.12±3.13 IU/L for Tubercular meningitis group. It was significantly higher (p<0.001) as compared to partially treated pyogenic meningitis (5.39±2.70 IU/L) and aseptic meningitis (1.92±0.56 IU/L) groups. A combination of clinical criteria along with biochemical test of Adenosine deaminase and C-reactive protein in CSF increased the sensitivity of diagnosing Tubercular meningitis and differentiating it from other forms of meningitis at an early stage.     

Usefulness of ferroxidase activity of ceruloplasmin in the diagnosis of Wilson’s disease

Serum ceruloplasmin is one of the most commonly used screening tests for Wilson’s disease. However immunological assays for ceruloplasmin are not recommended for diagnosis and management of Wilson’s disease through calculation of free copper index. Enzymatic methods using non-physiological substrates have toxicity and stability problems, making them difficult to automate. Ferroxidase assays may be a satisfactory alternative for measuring serum ceruloplasmin. The o-dianisidine hydrochloride manual method for estimation of serum ceruloplasmin enzyme activity was compared with an automated method using the ferroxidase activity of ceruloplasmin in measurement in a double blind study in 91 consecutive patients screened for Wilson’s disease. The o-dianisidine and ferroxidase methods both successfully identified 7 patients with Wilson’s disease. Values for these 7 patients in the o-dianisidine and ferroxidase methods were median 5.0 (range 0–16.0 U/L) and median 45.0 (range 4–166 U/L) respectively. There were 7 other positive values (<62 U/L) with the o-diansidine method and 2 (<200 U/L) with the ferroxidase method, where WD was not confirmed. ROC curves for both methods showed area under the curve of 0.998 for o-dianisidine and 0.997 for ferroxidase. Using literature cut off values of 62 U/L and 200 U/L respectively both methods had 100% sensitivity and specificity was 91.7% (o-dianisidine) and 97.6% (ferroxidase). For the o-dianisidine assay, specificity was improved to 98.8% using a cut off of 22.5 U/L. In the 84 persons (46 adults and 38 children) in whom the diagnosis of Wilson’s disease was not established, the mean value for ceruloplasmin activity by the o-dianisidine and ferroxidase methods was 124.7 ± 48.7 U/L and 571.4 ± 168.1 U/L respectively. There were no significant differences between sex or age of patients (p > 0.29). In a subsequent evaluation with 372 specimens, the Pearson correlation coefficient between the assays was 0.908, p < 0.01, slope 4.06, intercept 265.8, with the manual assay as the x-axis. The ferroxidase assay is a suitable replacement for the o-dianisidine assay in detecting patients with Wilson’s disease.     

Role of oxidative stress and antioxidants in aetiopathogenesis and management of oral submucous fibrosis

Lipid peroxidation product, malonaldehyde (MDA) and antioxidants were estimated in plasma and erythrocytes of 34 cases of oral submucous fibrosis (OSMF) of different grades with equal number of healthy controls to evaluate the association of reactive oxygen species (ROS) and OSMF. While plasma MDA was found to be significantly higher in patients (3.3±0.4 nmole/ml, P<0.001) as compared to controls (2.4±0.5 nmole/ml), plasma beta carotene and vitamin E levels were found to be decreased significantly in patients (81.7±14.3 μg/100 ml, P<0.001; 9.3±0.9 mg/L, P<0.01 respectively) with respect to healthy controls (110±20.8 μg/100 ml and 10.1±1.2 mg/L). The decrease in beta-carotene and vitamin E was found to be more significant in OSMF grade II and III than in grade I. After 6 weeks of oral administration of beta-carotene and vitamin E, patients showed increase in plasma level of these two antioxidants along with decrease in MDA level associated with clinical improvement.     

Serum adenosine deaminase in carcinoma breast

Serum Adenosine Deaminase (ADA) activity was studied in 20 control cases and 32 patients with carcinoma breast. In control group the ADA activity ranged from 13–94 IU/L with a mean of 43.75±21.55 IU/L while in patients of carcinoma breast it was 16–95 IU/L with a mean of 47.46±22.54 IU/L, but these values were not statistically different. The ADA activity was also compared with other parameters in patients of carcinoma breast but the difference was not statistically significant in the various parameters like duration of disease, menopausal status, tumor size, hemoglobin level, total leucocyte count, absolute lymphocyte count, total serum proteins, serum albumin, histological grade of tumor and lymphocytic infiltration. The only significant difference in ADA activity was when N1 status of lymph node was compared with N2 status of lymph node.     

Preliminary studies on serum lipids, apolipoprotein-B and oxidative stress in xanthelasma

Serum lipid profile, apolipoprotein-B (apo-B), malondialdehyde levels(MDA) and superoxide dismutase (SOD) activity were assessed in 12 cases of xanthelasma with and without coronary artery disease (CAD)/hypertension (HTN) and results are compared with healthy controls. Dyslipidemia was found in 65% cases of xanthelasma as compared to 20% healthy controls. Xanthelasma patients had significantly high malondialdehyde (MDA) levels (p<0.01) and significantly decreased (p<0.05) SOD activity as compared to controls. Among xanthelasma patients, xanthelasma with CAD/HTN showed higher total cholesterol (236±32.7 vs 188±24.7 mg/dl), low density lipoprotein cholesterol (157±35.5 vs 113±16 mg/dl) and Apo-B (120.5±9.4 vs 114±19.2 mg/dl) levels as compared to xanthelasma without CAD/HTN. Results of our study indicate that xanthelasma patients with increased apo-B, MDA and decreased SOD need cardiovascular monitoring.     

Role of reactive oxygen species in pathogenesis of nephrotic syndrome

Nephrotic syndrome is the common chronic disorder characterized by alteration of permeability of the glomerular capillary wall, resulting in its inability to restrict the urinary loss of proteins. Nephrotic syndrome is characterized by heavy proteinuria, hypoalbuminemia, hyperlipidemia associated with peripheral edema. The molecular basis of glomerular permselectivity remains largely unknown. In recent years it has been proposed that Nephrotic syndrome is a consequence of an imbalance between oxidant and antioxidant activity. The present study was aimed to test that the reactive oxygen species are the mediators of excessive protein permeability and other complications of Nephrotic syndrome. For this 30 adults with Nephrotic syndrome were studied. The control group comprised 30 healthy adults matched for age. Serum levels of lipid peroxides, nitric oxide (NO⊙), α- tocopherol, ascorbic acid, erythrocyte superoxide dismutase activity, serum albumin, uric acid, cholesterol and plasma total antioxidant capacity were measured. Student’s ‘t’ test was applied for statistical analysis. There was a significant increase in lipid peroxide (1.58 ± 0.42 in controls, 3.64 ±1.3 in patients) (P<0.001) levels in study group as compared with controls. α-tocopherol (12.95 ± 1.04 in controls, 9.93 ± 1.43 in patients) (P<0.001), erythrocyte SOD activity(1.88 ± 0.9 in controls 1.07 ± 0.5 in patients) (P=0.01), serum albumin(4.06 ± 0.50 in controls, 3.04 ± 0.11 in patients) (P<0.001), and plasma total antioxidant capacity (847.33 ± 126.83 in controls, 684.00±102.94 in patients) (P<0.001) were significantly decreased. There was non-significant increase in uric acid (P>0.05), a non-significant decrease in NO⊙ (38.48 ± 15.47 in controls 37.47 ± 14.27 in patients) (P>0.05) and ascorbic acid levels ascorbic acid,( 0.95 ± 0.31in controls 0.79 ± 0.30 in patients) (P>0.05) in study group as compared with controls. Imbalance between oxidants and antioxidants may contribute to pathogenesis of proteinuria and related complications in nephrotic syndrome.     

Effect of low-dose omega-3 fatty acids substitution on blood pressure, hyperinsulinemia and dyslipidemia in Indians with essential hypertension: A pilot study

The present study comprised 100 patients of essential hypertension who were screened for fasting hyperinsulinemia, which was detected in 77% cases. Twenty such hyperinsulinemic cases were subjected to 4 weeks of dietary control phase followed by 6 weeks of omega-3 fatty acids substitution [either 0.6 g/d (group 1) or 1.2 g/d (group 2)]. The mean basal fasting plasma insulin levels were significantly higher (p<0.001) in patients of hypertension when compared to normal controls (126.51±80.36 and 19.35±12.61 μU/ml respectively). At the end of 4 weeks of diet control only, no significant change was observed in any parameter. After substitution of omega-3 fatty acid, a significant reduction of fasting plasma insulin levels in both group 1 (29%) and group 2 (22.8%) was observed (p<0.001). Significant reduction of systolic and diastolic blood pressure (16.4% and 25% respectively), serum cholesterol, triglycerides and low density lipoprotein was also noted in both groups (p<0.001), while high density lipoprotein increased by ∼8 mg% in both groups. Thus omega-3 fatty acid substituion in low dose along-with curtailment of dietary omega-6 fatty acid may be used as an adjunctive measure in the management of essential hypertension.     

Serum Urea:Albumin Ratio as a Prognostic Marker in Critical Patients With Non-Chronic Kidney Disease

Routine laboratory investigations play an important role in estimating the risk of mortality in intensive care unit (ICU) patients. The significance of urea:albumin ratio (UAR) in predicting the stay and mortality of ICU patients is not known. It is a retrospective study of patients admitted to ICU (n = 412) with non-chronic kidney disease (non-CKD). Receiver-operating characteristics (ROC) analysis for predicting mortality was carried out to find area under curve (AUC) and threshold levels. Analysis of survival probability was carried out by Kaplan–Meier method and Log-rank test. The AUC to predict mortality were 0.695, 0.767 and 0.791 for serum albumin, urea and UAR, respectively. The threshold levels for albumin, urea and UAR were 2.8 g/dL, 53 mg/dL, and 23.44 mg/g, respectively. The highest odds ratio (OR) of 9.75 to predict mortality at threshold level was observed for UAR, while OR were 7.0 and 3.62 for serum urea and albumin, respectively. The serum urea above and albumin below threshold level were associated with increase in ICU stay of >3 days but the highest OR of 4.73 to predict stay of >3 days was observed for UAR. Kaplan–Meier survival analysis shows significant (p < 0.001) difference at the threshold value of UAR. Serum urea and albumin are found to be an independent predictor for the mortality and stay; however an increased UAR value is the best parameter in predicting mortality and stay in ICU patients with non-CKD illness.     

Seminal plasma levels of 15-F2α-isoprostane, malondialdehyde and total homocysteine in normozoospermic and asthenozoospermic males

It has been proposed that oxidative stress plays an important role in male infertility. The aims of this study were to compare seminal plasma levels of 15-F2t-isoprostane (8-iso-PGF2α), malondialdehyde (MDA), and total (sum of free and bound) homocysteine (tHcy) from normozoospermic vs. asthenozoospermic men, and to examine the relationships between tHcy and lipid peroxidation products. The study was a case-control study with a simple random sampling. The case group was consisted of 15 asthenozoospermic males. This group was compared with 15 normozoospermic men. Seminal plasma levels of 15-F2α-isoprostane and tHcy were measured using commercially available enzyme immunoassay (EIA) kits. MDA levels were determined by the thiobarbituric acid (TBA) assay. The Mann-Whitney U test was used to compare two groups. Coefficients of correlation were calculated using Spearman’s correlation analysis. All hypothesis tests were two-tailed with statistical significance assessed at the p value <0.05 level. MDA levels were higher in asthenozoospermic subjects than in control subjects (0.72±0.06 μM vs. 0.40±0.06 μM; p<0.05). No differences were seen in 15-F2α-isoprostane levels in asthenozoospermic subjects and controls (65.00±3.20 pg/ml vs. 58.17±4.12 pg/ml; p>0.05). Interestingly, tHcy levels were to be slightly higher in asthenozoospermic subjects than in controls (6.18±1.17 μM vs. 4.8±0.52 μM). Sperm motility was inversely correlated with seminal plasma 15-F2α-isoprostane and MDA levels, respectively (p<0.05). In summary, seminal plasma levels of 15-F2α-isoprostane and tHcy showed no significant difference between normozoospermic and asthenozoospermic men. Sperm motility was not correlated with seminal plasma levels of tHcy. No relationship was found between tHcy and lipid peroxidation.     

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.     

Assessment of Insulin Resistance and Metabolic Syndrome in Drug Naive Patients of Bipolar Disorder

The levels of fasting glucose, fasting insulin, insulin resistance (IR) and the prevalence of metabolic syndrome (MS) in a sample population of bipolar disorder (BPD) patients who were newly diagnosed and psychotropically naïve were assessed and compared with an age, sex and racially matched control population. 55 BPD-I patients (15–65 years) who were non-diabetic, nonpregnant, and drug naïve for a period of at least 6 months were included in the study. Diagnosis was made using the structured clinical interview for DSM-IV axis I disorders (SCID IV). IR was assessed using homeostasis model of insulin resistance (HOMA-IR); MS was defined according to National Cholesterol Education Program-Adult Treatment Panel III (NCEP-ATP III). Data were compared with 25 healthy controls. BPD patients had significantly higher mean levels of fasting plasma insulin (13.2 ± 9.2 vs. 4.68 ± 3.1 μIU/ml, p < 0.05), postprandial plasma insulin (27.2 ± 14.5 vs. 18.1 ± 9.3 μIU/ml, p < 0.05) and a higher value of HOMA-IR (3.16 ± 2.2 vs. 1.19 ± 0.8, p < 0.05) when compared to the controls. A significantly higher proportion of patients of BPD compared to controls were manifesting levels of fasting plasma glucose, serum triglyceride and blood pressure higher than the cut off while waist circumference and serum HDL cholesterol failed to show any significant difference in the proportion. There was a significantly higher proportion of prevalence of IR between BPD cases and controls (26/55 vs. 2/25, z value 9.97, p < 0.05) while there was no significant difference in proportion of prevalence of MS between these two groups. Within BPD patients, logistic regression analysis showed that age, sex or current mood status (depressed/manic) were not significantly predictive of presence or absence of MS or increased IR.     

Homocysteine status and acute myocardial infarction among Tamilians

Myocardial infarction is a major consequence of coronary artery disease. Apart from the traditional risk factors of myocardial infarction, recently many reports have suggested that hyperhomocysteinemia plays important role in myocardial infarction. Plasma homocysteine level was determined in 60 myocardial infarction patients and in 35 age matched healthy individuals. Statistically significant differences (p<0.01) were observed in the mean of plasma homocysteine concentrations between the acute myocardial infarction patients (24.59±6.14 mM/L) and in normal healthy individuals (13.73 ±3.54 mM/L). The level of homocysteine in myocardial infarction patients is significantly high (p <0.01) among myocardial infarction patients when compared to that of the controls. The the present study indicates a strong association between plasma homocysteine and acute myocardial infarction among Tamilians, thus implying plasma homocysteine as a possible risk factor for myocardial infarction.     

Changes in glycated proteins during impaired glucose tolerance

In persons with impaired glucose tolerance, serum glycated proteins (9.44±1.46μg hexose/mg protein) increased 1.69-fold (P<0.001) over control subjects compared to an increase of 1.95-fold in overt diabetics. However, correlations between serum glycated protein values and 2 h blood sugar values during the tolerance test (r=0.39) and between the former and graph area (r=0.30) were not strong. Glycated albumin (2.25 mol hexose/mol protein) and glycated haemoglobin (0.398±0.07 mol hexose/mol protein) in the study group also showed 1.67 and 1.24-fold increase respectively compared to controls. Fairly good correlations between glycated serum protein and glycated albumin (r=0.50) and glycated haemoglobin (r=0.45) were observed during this study. The data suggest that measurement of glycated proteins will be a useful index to detect impaired glucose tolerance.     

Blood lipid peroxidation products and antioxidants in senile cataract

Lipid peroxidation products and antioxidants were estimated in either blood, serum or erythrocytes of 25 cases of early and 25 of advanced senile (50–60 years) cataract and 30 persons of age matched healthy controls. Serum lipid peroxidation products (as malondialdehyde) were significantly higher in patients with early senile (0.25±0.05 μm/dl, P<0.05) and advanced senile cataract (0.29±0.05 μm/dl, P<0.001) as compared to healthy controls (0.22±0.07 μm/dl). Erythrocyte superoxide dismutase levels were lower in patients with early senile (730±60.5 units/gm Hb) and significantly lower in advanced senile (712±50.2 units/gm Hb, P<0.05) cataract than those in healthy controls (767±59.5 units/gm Hb). Serum α-tocopherol was significantly lower (P<0.05) in only advanced senile cataract (0.69±0.08 mg/dl) but not in early senile cataract (0.75±0.08 mg/dl) when compared with healthy controls (0.75±0.09 mg/dl). There was no significant difference in the erythrocyte concentration of reduced glutathione and serum levels of total proteins, β-carotene, vitamin A, ascorbic acid, calcium, magnesium and zinc.     

Diagnostic Value of Lactate Levels in Acute Abdomen Disorders

The supply of oxygen is limited in certain intra abdominal conditions due to direct vascular invasion or inflammatory process, resulting in high lactate levels. Aim of this study was to find the predictive value of lactate levels in the peritoneal fluid (PF) and blood of patients with acute abdomen. The study comprised of fifty patients with acute abdominal conditions, admitted in emergency ward of tertiary care hospital, thirty patients were with surgical abdomen (group I) and twenty patients with non surgical abdomen (group II). Lactate was estimated in PF and blood on Blood Gas Analyzer (NOVA, M-7). The mean lactate levels in PF were significantly higher in group I as compared to group II (14.65 ± 1.195 vs. 5.92 ± 0.97 mmol/L, p < 0.001). There was no significant difference in blood lactate levels in both the groups. When PF and blood lactate levels were compared within groups, we found that PF levels were significantly higher than blood in group I (14.65 ± 1.195 vs. 3.85 ± 0.54 mmol/L, p < 0.001) but not in group II (5.92 ± 0.97 vs. 4.36 ± 0.95 mmol/L). Diagnostic value was obtained using ROC curve. Cut off values obtained for PF lactate, difference and ratio of PF and blood lactate (≥6.4 mmol/L, ≥3.3 and ≥2.1 respectively) are at very high degree of sensitivity and specificity. So it can be useful marker of surgical emergency in patients with acute intra abdominal pathology, especially in clinically ill patients or in whom physical examination is not yielding because of neurologic disorders or unresponsiveness.     

Evaluation of oxidative stress, antioxidants and prolactin in infertile women

The objective of the study was to estimate the serum concentrations of oxidative stress markers-lipid hydroperoxide (MDA-Malondialdehyde), lactate dehydrogenase (LDH), nitrite; and ferric reducing antioxidant power (FRAP) — a measure of total antioxidant capacity in serum. Serum nitrite levels were high in fertile women than in infertile women (p<0.001), whereas MDA was high in infertile women than in fertile group (p<0.018). On classifying the entire group of women with infertility based on the etiology, serum MDA and LDH levels were higher in women with infertility due to unexplained factor than in women with other causes contributing to infertility compared to controls (p<0.001, p<0.02). A positive correlation among serum prolactin and nitrite suggested that hyperprolactinemia could contribute to infertility by inducing oxidative damage. Serum FRAP levels showed no significant difference in the groups studied. Lipid peroxidation was high in women with unexplained infertility which may be due to hyperprolactinemia.     

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.     

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.     

Use of specific IgE to fractions of gynandropsis gynandra pollen inin vitro diagnostic test

The study was aimed at presence of specific IgE antibody levelsinvitro to the identified antigen. Based on positive skin test with Gynandropsis gynandra and elevated levels of total IgE (>325 IU/ml) 104 patients were selected. Healthy, asymptomatic individuals (25) with low total IgE (<325 IU/ml) were included as controls. The mean OD values by ELISA for specific IgE were 0.67±0.21, 0.57±0.18 and 0.56±0.18 with whole pollen antigen, 46-37 kD fraction and 36-32 kD fraction, respectively. The specificity and sensitivity between skin test positivity with whole pollen antigen verses fraction with mol.wt 46-37 kD was 90% and 90% and for fraction with mol.wt 36-32 kD was found to be 81.1% and 89.4%. The clusters with molecular weights 46-37 kD and 36-32 kD may be useful inin vitro diagnostic test. Fractions within these clusters need to be identified for a higher specificity.