Salivary proteolytic activities in periodontitis, gingivitis and diabetes mellitus

Elastase activity was found to be significantly increased in periodontitis (0.872±0.4270 absorbance units/mg protein, mean±S. D., 1.05±0.61 units/ml saliva), gingivitis (0.772±0.416 units/mg protein, 1.515±0.952 units/ml) and diabetes (0.549±0.286 units/mg protein, 1.20±0.769 units/ml) compared to normals (0.255±0.089) units/mg protein, 0.264±0.079 units/ml). Chymotryptic activity was not found to be increased in these disease conditions over the normal level (0.284±0.096 units/mg protein). Aminopeptidase activity was elevated only in periodontitis (0.670+0.140 units/mg protein) compared to normals (0.349±0.100 units/mg protein). Trypsin-like activity was also found to be significantly raised in periodontitis compared to normals when Pro-Phe-Arg-p-nitroanilide (0.666±0.204 units/mg protein), Ile-Pro-Arg-p-nitroanilide (1.59±0.260 units/mg protein) and Pyroglu-Pro-Arg-p-nitroanilide (1.82±0.380 units/mg protein) were used as substrates. The normal values with these three substrates were respectively, 0.454±0.110, 1.04±0.231 and 1.15±0.312 units/mg protein. Total protein level in saliva was found to be significantly elevated in gingivitis (209±66.8 mg/dl) and diabetes (204±68.0) compared to normal values (107±20.7). Increase in periodontitis was marginal (127±28.3 mg/dl). Expression of proteolytic activities normalized to protein level was useful in differential diagnosis. Increase in elastase level in saliva is not a specific marker for periodontal diseases.     

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.     

Loss of proteinase binding capacity of plasma alpha2 macroglobulin in some disease conditions

Alpha₂-macroglobulin (MG) levels in terms of proteinase binding capacity in fresh plasma of diabetics (males 183.9±60.3 mg MG/dl, females 173.8±49.7) and viral hepatitis patients (195.2±41.4) were comparable to normal sujbects (males 180.0±28.6, females 213.0±59.6) whereas in renal failure the values were decreased (144.0±30.0) and was found to be increased in nephrotic syndrome (331.5±47.9). MG activity was stable in normals and nephrotic patients in plasma on standing for 12 hr. at 37°C. In other conditions, the proteinase binding capacity was considerably reduced (mean decrease: 51.2% in diabetes, 64.0% in renal failure and 52.6% in viral hepatitis). Concentration of MG determined by immunoelectrophoresis did not however change in diabetic plasma on 12 hr. incubation. It is proposed that proteinases are continously generated in plasma in some disease conditions which bind to MG causing reduction in measured proteinase binding capacity.     

Changes in levels of serum glycoproteins in major depressive disorders

The present study deals with estimation of levels of fractions of serum glycoproteins, protein bound hexose (PBH), protein bound hexosamine (PBHex), protein bound fucose (PBF), protein bound sialic acid (PBS) and protein bound carbohydrate (PBC) in thirty patients of Major Depressive Disorders (MDD) in comparison with thirty normal subjects. In patients of MDD, the level of PBH, PBHex, PBF, PBS and PBC were significantly higher as compared to the normal subjects (p<0.05). In patients, of MDD, after one-month treatment with fluoxetine, the levels of PBH, PBHex, PBF, PBS and PBC were significantly decreased as compared to the levels of these fractions in same patients of MDD before beginning of the treatment (p<0.05). Based on findings of the present study, it can be concluded that changes in the level of serum glycoproteins level before and after treatment with fluoxetine can be correlated with clinical status of MDD.     

Protective role of<Emphasis Type="Italic">Phyllanthus niruri</Emphasis> against nimesulide induced hepatic damage

Present study aimed to evaluate the protective role of the aqueous extract of Phyllanthus niruri (P. niruri) against nimesulide-induced hepatic disoder in mice by determining levels of glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and alkaline phosphatase (ALP) in serum and also by measuring the hepatic content of the antioxidant enzymes, superoxide dismitase (SOD) and catalase (CAT); the free radical scavenger, reduced glutathione (GSH) and thiobarbituric acid reacting substances (TBARS). Aqueous extract of P. niruri was administered either orally or intraperitoneally in different doses and times as needed for the experiments. Intraperitoneal of the extract (100 mg/kg body weight for seven days) reduced nimesulide (750 mg/kg body weight for 3 days) induced increased levels of GOT (37.0±1.8 units/ml in control group vs. 91.8±2.0 units/ml in nimesulide treated group vs. 35.0±1.0 units/ml in extract treated group), GPT (30.0±2.1 units/ml in control group vs. 88.4±2.9 units/ml in nimesulide treated group vs. 34.1±1.8 units/ml in extract treated group), and ALP (7.86±0.47 KA units/ml in control group vs. 23.80±0.60 KA units/ml in nimesulide treated group vs. 7.30±0.40 KA units/ml, in extract treated group) to almost nomal. In addition, P. niruri restored the nimesulide induced alterations of hepatic SOD (550±20 units/mg total protein in control group vs. 310±13 units/mg total protein in nimesulide treated group vs. 515±10 units/mg total protein in extract treated group), CAT (99.5±2 units/mg total protein in control group vs. 25.0±1.5 units/mg total protein in nimesulide treated group vs. 81.0±0.8 units/mg total protein in extract treated group), GSH (90±3 nmoles/mg total protein in control group vs. 17±4.2 nmoles/mg total protein in nimesulide treated group vs. 81±1 nmoles/mg total protein in extract treated group) and TBARS (measured as MDA, 36.6±3.0 nmoles/g liver tissue in control group vs. 96.3±5.2 nmoles/g liver tissue in nimesulide treated group vs. 41.2±1.7 nmoles/g liver tissue in extract treated group) contents. Dose-dependent studies showed that the herb could protect liver even if the nimesulide-induced injury is severe. Intraperitoneal administration of the extract showed better protective effect than oral administration. Combining all, the data suggest that P. niruri possesses hepatoprotective activity against nimesulide-induced liver toxicity and probably acts via an antioxidant defense mechanism. To the best of our knowledge, this is the first report of the hepatoprotective action of P. niruri against nimesulide induced liver damage.     

Blood zinc protoporphyrin, serum total protein, and total cholesterol levels in automobile workshop workers in relation to lead toxicity: Our experience

Blood zinc protoporphyrin (ZPP), serum total protein (TP), and total cholesterol (TC) levels in automobile workshop workers in relation to lead toxicity were analysed. In the present study, automobile workshop workers (healthy male workers at an age between 28 and 35 from four major automobile workshops in Kottayam, Kerala State, India) and the control (male healthy adults at an age between 28 and 35 residing at Aymanam, a distant village at Kottayam District, Kerala having reduced or no chance of lead exposure) displayed significant difference in blood lead (BPb) and blood ZZP (BZPP) level. The mean value of BPb in automobile workshop workers was 15.76±0.33 μg/dl, while in the control it was 8.20±0.15 μg/dl. In automobile workshop workers, the mean value of BZPP was 34.2±0.62 μg/dl. The control group exhibited a mean of 11.5±0.22 μg/dl. Automobile workshop workers exhibited significant increase in BZPP was corresponding to the increase in BPb level. The total protein levels estimated in automobile workshop workers showed significant decrease compared to control individuals, but was within the reference range of healthy individuals. The mean value of TP level in automobile workshop workers and control was 6.9±0.13 g/dl and 7.71±0.18 g/dl, respectively. There was no significant difference in blood haemoglobin (BHb) level among the automobile workshop workers and control. The serum TC level in automobile workshop workers showed significant decrease compared to the control individuals, but was with in the reference range of healthy individuals. The mean level of serum TC in automobile workshop workers was 162.00±3.44 mg/dl and the same in control was 172.86±4.32 mg/dl. The present study affirms occupational lead toxicity in automobile workshop workers and its effect on serum protein and cholesterol levels.     

Hesperetin on Cell Surface Glycoconjugates Abnormalities and Immunohistochemical Staining with Cytokeratin in 7,12 Dimethylbenz(a)anthracene Induced Hamster Buccal Pouch Carcinogenesis

Hesperetin, a naturally occurring citrus flavanone of the bioactive substance, possesses different pharmacological and biochemical activities including anti-cancer and anti-oxidants effect. The aim of the study to investigate that hesperetin on abnormalities of glycoconjugates (protein bound hexose, hexosamine, total sialic acid and fucose), histology (PAS staining) and immunoexpression of cytokeratin during 7,12-dimethylbenz(a)anthracene (DMBA) induced hamster buccal pouch (HBP) carcinogenesis. Oral tumors were developed in the buccal pouches of male golden Syrian hamsters by topical application of 0.5% DMBA thrice a week for 10 weeks and developed morphological alterations depicted as hyperplasia, dysplasia and well-differentiated squamous cell carcinoma formation with noticeable abnormalities of glycoconjugates and cytokeratin. The protective effect of hesperetin against DMBA was evaluated by assessing immunohistochemical expression, histological sections of buccal tissues and the levels of glycoconjugates in the buccal mucosa and plasma were analyzed. Hesperetin administrated orally at a dose of 20 mg/kg b.w. to hamsters treated with DMBA, significantly reduced the status of glycoconjugates and cytokeratin to the near normal range. Overall findings accomplished that hesperetin protects cell surface glycoconjugates abnormalities in DMBA induced HBP carcinogenesis.     

Serum Levels of Glycoproteins are Elevated in Patients with Ovarian Cancer

Identification of reliable biomarkers for detection and staging of cancer and monitoring the outcome of anticancer therapy has been considered to be of high importance. We aimed to estimate the levels of serum glycoproteins, protein bound-hexose, protein bound hexosamine, protein bound fucose, protein bound sialic acid and protein bound carbohydrate in 32 ovarian cancer patients and compared them with the levels that found in 25 normal subjects. As compared to the normal subjects, all the four fractions of glycoproteins level were significantly elevated in ovarian cancer patients (p < 0.05). Chemotherapy in these patients significantly decreased the levels of serum glycoproteins (p < 0.05). Thus, high levels of serum glycoproteins in ovarian cancer patients could be due to abnormal protein glycosylation indicating malignant transformation of the cells.     

Plasma proteins and proteinuria in gestational malaria

The plasma concentrations of total protein, albumin, immunoglobulins IgG, IgA and IgM and urinary protein were assayed in 250 pregnant Nigerian women with malaria and compared with 250 healthy pregnant women which served as controls. The mean values of plasma total proteins, albumin, IgG and IgA were significantly lowered (P<0.05) while a slight increase in IgM was observed in the malaria patients. Urinary proteins value of 23.10±0.50 mg/dl was obtained for the pregnant women with malaria, this was significantly higher (P<0.05) than that of the controls with the corresponding value of 15.32±0.09 mg/dl. This study has therefore demonstrated elevations of the urinary and decrease in plasma proteins in gestational malaria. These findings suggest that the protein profile should be considered in the diagnosis and treatment of malaria.     

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.     

Value of ascitic fluid cholesterol and serum-ascites albumin gradient in differentiating cirrhotic and malignancy related ascites

In a perspective study, the ascitic fluid and serum concentration of total cholesterol, total proteins and albumin in a group of 45 patients was studied. Patients with nonmalignant or cirrhotic ascites were compared with patients having malignancy related ascites and it was proved that the ascitic fluid cholesterol and the serum ascites albumin gradient helped to differentiate cirrhotic from malignant ascites. These two parameters showed a remarkable relationship to the presence/absence of malignancy. Non malignant ascites patients had ascitic fluid cholesterol values of 19.41±8.33 mg/dl, as against the malignancy related ascites patients, who showed levels of 95.87±1.24 mg/dl. Similarly, the serum-ascites albumin gradient levels were 2.89±0.65 in non malignant ascites patients, while the malignancy related ascites cases had 0.86±0.50. The discrimination values for cholesterol were taken as 45 mg/dl while that for serum ascites gradient was taken as 1.1. Levels of serum cholesterol, total protein and albumin were not significantly altered.     

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.     

Serum and urinary zinc in zinc miners and zinc smelters

Serum (200.6±64.8 mg/dl) and urinary zinc (1186.0±265.5 mg/l.) of zinc miners at Zawar were significantly higher as compared to zinc smelter workers (Serum 89.7±9.9. mg/dl; Urine 590.0±32.1 mg/l.). Debari and Udaipur residents (Serum 104.0±18.6 mg/dl; Urine 735.0±180.2 mg/l.). This was most likely attributable to higher dietary zinc intake by zinc miners due to higher zinc content of foodstuffs grown in Zawar areas as compared to Debari and Udaipur.     

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.     

Serum lipids in dilated cardiomyopathy

Serum total cholesterol and triglyceride levels were estimated in 45 dilated cardiomyopathy (DCM) patients and were compared with 31 healthy controls. The total serum cholesterol (215±50 mg/dl) and serum triglyceride (162±76 mg/dl) levels in DCM patients did not show any significant changes as compared to normal controls. The cholesterol and triglyceride and the individual lipoprotein fractions, viz HDL, VLDL, LDL and chylomicrons were analysed in 10 patients. The values did not differ significantly when compared to those of controls. These findings suggest that serum lipids may not play a role in the antietiopathogenesis of DCM.     

Evaluation of reference intervals of serum lipid profile from healthy population in Western Maharashtra

Fasting samples of 914 subjects from healthy population were analyzed for total cholesterol, triglyceride and three major fractions of lipoproteins i.e. high-density lipoprotein cholesterol, low lipoprotein cholesterol and very low-density lipoprotein cholesterol. The values obtained were (in mg/dl) 165.7±30.2,88.36±31.2, 44.86±10.68, 101.66±29.8 and 18.11±7.35 respectively. When these subjects were grouped according to the age and sex, no appropriate differences were observed between most of the groups. Triglycerides were found to be low and HDL cholesterol was high in female when compared with male of similar age. Beyond age 40 years cholesterol level and low density lipoprotein cholesterol was found to be gradually increased in case of women. Minor difference was observed with dietary pattern. Present study suggests that clinical evaluation of patient should be made on the basis of these reference values for Western Maharashtra population.     

Metabolic alterations during Ramadan fast

Some biochemical parameters in 12 young Muslim students who underwent fasting during the entire one month period of Ramadan were investigated. There was a significant reduction in their body weight (72.5±4.0 to 69.4±3.0 kg) and blood glucose levels (99.6±3.5 to 82.9±7.3 mg/dl). Though the morning plasma cortisol fell (215.2±22.1 to 105±11.2 μg/l) and the evening plasma cortisol rose (58.4±5.1 to 64.4±4.8 μg/l) significantly there was no reversal of the circadian rhythm of cortisol secretion. A decline in the diastolic blood pressure (80 to 75±6 of Hg) of these individuals though not significant was also noted. However, cholesterol (160.9±12.8 to 178.2±40.6 mg/dl) and HDL-Cholesterol (42.6±2.2 to 48.1±2.4 mg/dl) showed a tendency for increase. All the changed values were within the normal range.     

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.     

Serum glycoconjugates and ceruloplasmin in cancer of uterine cervix

The present study was done to assess the levels of glycoconjugates and ceruloplasmin in sera of patients with cervical cancer. Serum hexoses, hexosamines, sialic acid and fucose are elevated in a variety of inflammatory and neoplastic conditions. All the glycoconjugates, except fucose were increased in serum of patients compared to controls. Also, hexoses and sialic acid levels were high in patients with later stages of cancer compared to patients with early stage disease (P=<0.0001, P=0.03). Serum ceruloplasmin was increased in patients with early stage cancer (51.5mg/dl) and with late stage cancer (61mg/dl) compared to controls (38mg/dl). The elevated glycoconjugates may be the result of inflammatory reaction associated with neoplasia, as serum ceruloplasmin (an acute phase reactant) is also increased in these patients.     

Pleural fluid MDA and serum-effusion albumin gradient in pleural effusion

Pleural fluid malondialdehyde (PMDA) and serum effusion albumin gradient(SEAG) were estimated in 60 patients of pleural effusion of diverse etiologies. The results were compared with Light’s criteria to distinguish between transudates and exudates. The mean PMDA level was 0.68±0.24nmol/ml and 1.17±0.25nmol/ml in transudates and exudates respectively showing a statistically significant (p<0.05) rise in exudates in comparison to transudates. SEAG registered a significant fall in exudates (P<0.001) when compared with transudates. PMDA revealed a positive correlation with pleural protein(r=+0.30) and a significant negative association with SEAG (r= −0.33).Sensitivity and specificity of PMDA were better than the parameters of Light’s criteria. Whereas SEAG documented approximately equal sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) with Light’s criteria. Therefore PMDA and SEAG can be taken together in addition to Light’s criteria to strengthen the discrimination between transudates and exudates in borderline cases of pleural effusion.