Association between Nutritional Status and Early Childhood Caries in Indian Children.

AIMS AND OBJECTIVES: To assess the body mass index (BMI) status and to correlate between nutritional status and early childhood caries among 3-6-year-old children in Vikarabad, Telangana, India. MATERIALS AND METHODS: A cross-sectional study was conducted among 350 children attending the outpatient department of Pedodontia in Sri Sai College of Dental Surgery, Vikarabad. Caries experience was recorded using dmft index (WHO 1997); various anthropometric measures, such as weight, BMI, and height, were recorded as per the standard guidelines. Pearson correlation coefficient and chi square test were used to determine significant differences. Data were analyzed using the statistical analysis software (SPSS version 17.0 Chicago IL, USA). RESULTS: Maximum number of participants (69%) were with normal BMI- for-age followed by underweight, overweight, and obese categories. The mean dmft values for each BMI category among the underweight, normal, overweight, and obese children were 4.9 ± 4.0, 4.6 ± 3.3, 2.7 ± 1.9, and 4.4 ± 3.0, respectively, which was statistically not significant. CONCLUSION: No significant association was found between dmft and BMI-for-age, rather a negative correlation existed between dmft and BMI-for-age (r = -0.023), which was not statistically significant (P = 0.66).

Effect of norethisterone acetate on serum lipid and lipoprotein parameters as well as on blood coagulation in female monkeys (M. fascicularis).

The effects of daily oral administration of a high dose of 10 mg norethisterone acetate (NET-Ac.)/kg/day over 14 weeks on serum lipid and lipoprotein parameters as well as on blood coagulation were investigated in female monkeys (M. fascicularis). Measurements of lipids and lipoprotein cholesterol were performed in weeks--5 and -1 before treatment and in weeks 4, 8 and 12 after treatment. In addition, various blood coagulation and fibrinolytic parameters were determined in weeks 11-14 after treatment with NET-Ac. Furthermore, the serum levels of norethisterone (NET) were determined in order to monitor the real systemic compound exposure and revealed that Cmax and AUC (0-3 h) values reached for norethisterone in this experiment in monkeys were about 25 times higher than those obtained after an oral contraceptive dose of NET-Ac. in women. The results of lipid and lipoprotein cholesterol determinations showed decreases in serum total lipids, phospholipids, triglycerides and total cholesterol associated with similar decreases in HDL-, LDL- and VLDL-cholesterol fractions after NET-Ac.-treatment in monkeys. These effects were observed from week 4 onwards and maintained their magnitude up to week 12 after treatment. Since both HDL- and LDL-cholesterol fractions decreased, the HDL/LDL-ratio remained almost unchanged. Thus, the results obtained in this study after high-dose treatment with NET-Ac. in monkeys did not indicate any changes of lipid and lipoprotein parameters which in humans are supposed to be associated with an increased risk of cardiovascular lesions, namely a decrease in HDL- and increase in LDL-cholesterol fractions. The results of blood coagulation and fibrinolytic parameters showed increased antithrombin-III and plasminogen levels besides minor changes in other parameters, thus indicating that NET-Ac.-treatment does not contribute to an increased risk of cardiovascular thrombotic events in the cynomolgus monkey.

Serum lipoprotein pattern in rats, dogs and monkeys, including method comparison and influence of menstrual cycle in monkeys.

The serum lipoprotein pattern (HDL-, LDL- and VLDL-cholesterol) was determined in normal untreated rats, dogs and monkeys using the density gradient centrifugation method, and compared with that in humans. The serum of rats (Wistar) and dogs (Beagle) was found to contain mainly HDL-cholesterol, whereas the serum of monkeys (Macaca fascicularis) consists of similar amounts of HDL-cholesterol and LDL-cholesterol, as in humans. In contrast to rats and dogs, monkeys showed a very low amount of serum VLDL-cholesterol. Comparison of four different methods (density gradient centrifugation, FPLC, agarose gel electrophoresis and precipitation methods) for determination of HDL- and LDL-cholesterol in monkey serum showed that density gradient centrifugation and FPLC-methods have the highest accuracy and recovery rate. The FPLC-method offers the extra advantages that it requires less sample volume and is less laborious. In a separate experiment, the influence of the menstrual cycle on the serum lipoprotein pattern was investigated in female monkeys. No changes in total cholesterol, HDL- or LDL-cholesterol were detected in serum samples obtained in the follicular and luteal phases of the cycle. In conclusion, monkeys (Macaca fascicularis) may be more suitable than rats or dogs for the comparison of the effect of different compounds on serum lipoproteins in pharmacological and toxicological studies, because the lipoprotein pattern in this species is similar to that of humans.

Concentrations in serum of IgG, IgM and IgA and their age-dependence in beagle dogs as determined by a newly developed enzyme-linked-immuno-sorbent-assay (ELISA).

The concentrations of immunoglobulins IgG, IgM and IgA and their age dependence were determined in the serum of normal, untreated beagle dogs by a newly developed sandwich enzyme-linked-immuno-sorbent-assay (ELISA). A clear age-dependent increase between approximately 0.8 and 1.6 years of age was observed for the immunoglobulin IgA, whereas IgG and IgM showed only a slight tendency to an age-dependent increase. For immunotoxicological characterization of various compounds especially in long term studies, this IgA age-dependence has to be considered in the planning and interpretation of studies with beagle dogs.

Effect of three non-ionic contrast media on rats and rabbits with regard to renal changes. Interspecies comparison.

The effect of three non-ionic contrast media (iosimide, iopamidol and iopromide) was investigated in rabbits and rats after single i.v. application up to high doses with regard to renal changes. Determinations of serum urea nitrogen and creatinine and urinary enzymes (lactate dehydrogenase, gamma-glutamyl-transpeptidase and N-acetyl-beta-D-glucosaminidase) as well as histological examinations of the kidneys were performed. The results obtained demonstrate that rabbit as an experimental animal was more sensitive than rat in exhibiting renal changes to contrast media. Furthermore, determinations of urinary enzymes demonstrated that gamma-GT in rabbits and LDH in rats were the most sensitive indicators for detection of early kidney damage. With regard to the high dose levels required, these kidney changes do not indicate any risk to humans at diagnostic dose levels.

Effect of iloprost on serum creatine kinase and lactate dehydrogenase isoenzymes after isoprenaline-induced cardiac damage in rats.

The effect of iloprost was investigated in an experimental model of cardiac damage in rats after isoprenaline (isoproterenol) treatment. Iloprost was administered by continuous subcutaneous infusion at a dose of 0.44 micrograms/kg body weight (b.w.)/min over a total period of 9 days. On day 8, 5 mg isoprenaline/kg b.w. was applied subcutaneously to induce the cardiac damage. The determinations of creatine kinase (CK, EC 2.7.3.2) and CK isoenzymes, lactate dehydrogenase (LDH, EC 1.1.1.27) and LDH isoenzymes as well as a-hydroxybutyrate dehydrogenase (a-HBDH, no EC) were performed 2, 6 and 24 h after isoprenaline application. Immediately after the last blood sampling, the animals were sacrificed and the hearts were examined histologically. Iloprost-pretreated animals showed a reduction in the rise in heart-specific isoenzymes CK-MB and LDH1-3 in the serum after isoprenaline application. A decrease in isoenzymes CK-BB and LDH4-5 in the serum was also observed in iloprost-pretreated rats. However, no difference could be detected histologically in the extent of myocardial necrosis between animals treated with isoprenaline alone or in combination with iloprost. These results suggest a biochemical cardioprotective effect of iloprost in this rat model after isoprenaline application and the lack of correlation with histological findings is discussed.

The use of urinary N-acetyl-beta-D-glucosaminidase (NAG) for the detection of contrast-media-induced 'osmotic nephrosis' in rats.

Excretion of urinary N-acetyl-beta-glucosaminidase (NAG), lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and gamma-glutamyltransferase (gamma-GT) was studied following single intravenous administrations of a non-ionic monomeric contrast medium (iohexol) at doses of 5.0, 7.5, 10.0 and 12.5 g iodine/kg body wt in rats. Measurements of urinary enzymes, serum urea nitrogen and serum creatinine were carried out on the 2nd, 3rd, 4th and 8th days after treatment. Histological examinations of kidneys were performed on days 3 and 8. From 5.0 g iodine/kg onwards urinary NAG showed a dose-dependent and significant (multiple comparison, alpha = 0.05) increase in the first 18-h urine samples after application. A significant increase in urinary LDH could be observed only at the highest dose of 12.5 g iodine/kg. All other biochemical parameters showed no differences when compared to the control group. The dose-dependent increase in lysosomal NAG correlated with the histological findings, i.e. there was dose-dependent vacuolization of proximal tubular cells, so-called 'osmotic nephrosis'.

Serum isoenzyme pattern of creatine kinase and lactate dehydrogenase in various animal species.

The creatine kinase and lactate dehydrogenase isoenzyme pattern were determined in the serum of normal and untreated rats, rabbits, dogs, monkeys and pigs. The relative distribution of all isoenzymes in the serum and an electrophoretic pattern for each animal species are presented. The isoenzyme serum pattern showed a great variation between the species. The diagnostic value of serum creatine kinase isoenzyme MB and lactate dehydrogenase isoenzymes 1 and 2 in predicting cardiac lesions in different animal species is briefly discussed.

Diagnostic value of serum creatine kinase and lactate dehydrogenase isoenzyme determinations for monitoring early cardiac damage in rats.

Serum activities of creatine kinase (CK, EC 2.7.3.2) and CK isoenzymes, lactate dehydrogenase (LDH, EC 1.1.1.27) and LDH isoenzymes, alpha-hydroxybutyrate dehydrogenase (alpha-HBDH, no EC) and the LDH/alpha-HBDH ratio were studied following a single s.c. application of 5-250 mg isoproterenol/kg body weight (b.w.) in rats. Measurements of the serum enzymes and histological and enzyme-histochemical examinations of hearts were performed 2, 4, 6, 8 and 24 h after treatment. A drastic increase in serum levels of the isoenzymes CK-MB, LDH1, LDH2 and alpha-HBDH and decrease in the ratio LDH/alpha-HBDH were observed from 2 h onwards after isoproterenol application in all dose groups, the maximum effect being after 4-8 h. Focal cellular injury in the myocardium could also be observed from 2 h onwards after isoproterenol application by an enzyme-histochemical method using nitro blue tetrazolium (NBT) whereas the earliest histological alterations using haematoxylin and eosin (HE) stain could be detected 6 h after treatment. A dose-dependent effect as to enzyme values as well as to myocardial necrosis was observed 24 h after isoproterenol application. No kidney damage could be detected on the basis of serum urea nitrogen and histological examinations. Thus, measurement of serum activities of CK-MB isoenzyme alone or LDH1-2 isoenzymes in combination with other tests to exclude kidney damage are valuable indicators of cardiac lesions in rats.

Value of enzyme determinations in urine for the diagnosis of nephrotoxicity in rats.

Excretion of urinary lactate dehydrogenase (LDH, EC 1.1.1.27), gamma-glutamyltransferase (gamma-GT, EC 2.3.2.2), alkaline phosphatase (ALP, EC 3.1.3.1), alanine aminopeptidase (AAP, EC 3.4.11.-), alanine aminotransferase (GPT, EC 2.6.1.2) and N-acetyl-beta-D-glucosaminidase (NAG, EC 3.2.1.30) was studied following a single i.v. application of 1 mg mercuric chloride/kg body weight or a radio contrast medium (SH H 340 AB) at a dose of 7.5 g iodine/kg body weight in rats. Measurements of urinary enzymes and serum urea nitrogen and creatinine were carried out on the second, third, fourth and ninth days after treatment. Histological examinations of kidneys were performed on day 9. A drastic increase in urinary LDH and moderate increase in gamma-GT, ALP and AAP and a very slight increase in GPT was observed in the first 18-h urine samples after mercuric chloride. This increase in enzymuria was associated with a drastic increase in serum urea nitrogen and creatinine, with a maximum on day 4. The radio contrast medium-treated animals showed a similar but less pronounced pattern of urinary enzymes excretion and only a slight increase of serum urea nitrogen on day 2. A good correlation was found between histological findings and enzymuria as well as serum urea nitrogen and creatinine. Thus, determination of only some urinary enzymes (LDH and gamma-GT) is valuable in predicting early nephrotoxicity and sufficient for the diagnosis of proximal tubule damage in rats.

Effect of cyproterone acetate and its metabolites on the adrenal function in man, rhesus monkey and rat.

The effect of oral administration of 17 alpha-acetoxy-6-chloro-1,2 alpha-methylene-4,6]pregnadiene-3,20-dione (cyproterone acetate, Androcur) on the adrenal function in male hypersexual subjects and in female rhesus monkey was investigated on the basis of cortisol and ACTH levels in serum or plasma and excretion of cortisol and 17-ketosteroids. In addition, cyproterone acetate and its main metabolite alcohol of the main metabolite were characterized for treatment of male hypersexual subjects and female rhesus monkeys did not reveal any signs of adrenal suppression. Cyproterone acetate and its metabolites gave on indication of any appreciable anti-inflammatory effect in the adjuvant edema test in rats. However, there was a general increase in the level of blood glucose and liver glycogen as well as a reduction in body weight and organ weight (spleen, thymus and adrenal) in rats, in which 15 beta-hydroxy cyproterone was slightly more active with the exception of adrenal weight tests. It can be concluded that adult man and rhesus monkey are much less sensitive, is so at all, to some corticosteroid-like activities of cyproterone acetate and its main metabolites than the rat.

Characterization of a new potent heparin. 6th communication: The comparison of the anticoagulant effect of the sodium and calcium salts of a new potent heparin after s.c. injection in beagle dogs.

The anticoagulant effect of the sodium and the calcium salts of a new potent heparin was compared in beagle dogs following a single s.c. injection. The determinations of whole-blood clotting time, activated partial thromboplastin time and plasma heparin levels using the amidolytic assay 1, 2, 4, 6, 8, 10 and 12 h post injection revealed that the calcium salt was just slightly more active than the sodium salt of heparin. The influence of the molecular composition in particular the inorganic ion content associated with heparin on the anticoagulation activity achieved after subcutaneous injection, is discussed.

Characterization of a new potent heparin. 7th communication: A comparative study of a new potent heparin in human after i.v. application.

The anticoagulant effect of a new potent heparin was compared with commercial heparin after a single i.v. application in human subjects. Determinations of thrombin time, whole-blood clotting time, activated partial thromboplastin time and plasma heparin levels using factor Xa inactivation assay were performed 5, 10, 15, 30 and 60 min after application. The new potent heparin was 1.5 to 2.0 times more effective than commercial heparin per mg dry weight depending upon the coagulation tests used and the length of observation period. In addition, determinations of biochemical and haematological parameters, 24 h after heparin application indicated no signs of any adverse effect.

Corticosteroid-like effects after high doses of compound (5-methoxy-2-thienyl)-thioacetic acid sodium salt in rats.

During a systemic tolerance study in rats with compound (5-methoxy-2-thienyl)-thioacetic acid sodium salt (MTTA), which is known to decrease the lipolytic process and platelet aggregation, effects after high doses were observed which are similar to the well-known findings after corticosteroid administration. The oral application of MTTA in a dose range from 10 to 1000 mg/kg over a period of 4 weeks resulted in a decrease in total lipids and free fatty acids from 100 mg/kg onwards and an increase in total glycerin and triglycerides after 1000 mg/kg, supplemented with an increased excretion of keto-bodies in urine at all dose levels. The hematological examinations revealed a reduction of blood eosinophils and lymphocytes in males at doses from 10 mg/kg, whereas the eosinophilic granulocytopoiesis was decreased only at the 1000 mg/kg level in both sexes. Determinations of organ weights showed a decrease in thymus weight from 300 mg/kg onwards with an involution at the highest dose levels, being confirmed by histological examination, whereas the adrenal glands' weights were decreased only at the dose level of 1000 mg/kg. Since the endogenous corticosterone level remained unaltered at all doses, it is suggested that these corticosteroid-like effects are directly attributable to MTTA.

Functional morphology of spontaneous hyperplastic and neoplastic lesions in the canine pituitary gland.

The pituitary glands of 10 male and 29 female dogs of different breeds had diffuse hyperplasia and hypertrophy of growth hormone or prolactin cells. Immunoreactive adrenocorticotrophin/melanotrophin was in the frequent focal hyperplastic lesions and micro- and macroadenomas of both pars distalis and pars intermedia of old female dogs. Pituitary tumours of other functional cell types were not found. Focal hyperplastic and neoplastic lesions usually were accompanied by spontaneous nodular hyperplasia or adenomas of the adrenal cortex as well as by changes in serum adrenocorticotrophic hormone and cortisol levels.

Effect of corticosteroids on intraocular pressure, ascorbic acid and sialic acid concentrations in aqueous humour and on mucopolysaccharides biosynthesis in the cornea of rabbits.

The effect of daily oral administration of corticosteroids on ocular dynamics and biosynthesis of corneal mucopolysaccharides was investigated using rabbits as experimental animals. Fluocortolone treatment at doses of 0.05 to 10 mg/kg resulted in a dose-dependent rise of intraocular pressure (IOP). At higher dose levels of 5 and 10 mg fluocortolonel kg daily over 14 days, an inhibition of 35S-incorporation and a reduction in the nexosamine content of the corneal mucopolysaccharides was observed. When the effects of prednisolone, fluocortolone and dexamethasone on IOP were compared the relative order of the potency for the corticosteroids was as follows: dexamethasone > fluocortolone > prednisolone. The influence of different doses of fluocortolone on ascorbic and sialic acid content in aqueous humour and plasma was also studied. Levels of 5 and 10 mg/kg caused a significant reduction of ascorbic acid and a slight increase in sialic acid content in aqueous humor besides an elevation of IOP, whereas the plasma levels of both components did not reveal any difference from control group. These findings suggest that the blood-aqueous barrier becomes more permeable under the treatment with corticosteroids, especially in doses which are known to be toxic in animals. Further, the rabbit can be used as an experimental animal for comparing the effets of different corticosteroids on IOP.

Characterization of a new potent heparin. 1 communication: Determinations of anticoagulant activity of a high potency heparin preparation in vivo using dog as an experimental animal and in vitro using dog and human plasma.

The anticoagulant effect of a highly potent heparin preparation was compared with a commercially available heparin in vivo after i.v. application in beagle dogs. The anticoagulant activity was determined using thrombin time, activated partial thromboplastin time and whole blood clotting time 5, 10 and 30 min after application. The relative potency of the heparin preparation (Schering) was found to be 1.62 to 2.52 times higher than heparin used for comparison (150 USP units/mg). The anticoagulant properties of both preparations were also studied in vitro using heparin concentrations from 0.44 to 7.0 microgram/ml dog and human plasma. The relative potency in vitro experiments using dog or human plasma were 1.62 and 1.63, respectively, on the basis of activated partial thromboplastin time. It was further demonstrated that the anticoagulant effect as determined by activated partial thromboplastin time in vitro on human plasma was 2 to 2.4 times more pronounced in both heparin preparations when compared to the effect exerted on dog plasma.

Characterization of a new potent heparin. 2nd communication: Chemical analysis of the carbohydrate content and determination of the biological activity of a new potent heparin preparation in vitro, using protamine neutralization and amidolytic methods for factor Xa and thrombin.

A new potent heparin preparation was further characterized for carbohydrate content and for biological activities in vitro using the protamine neutralization test and amidolytic methods for factor Xa and thrombin. The chemical analysis of the heparin preparations showed similar amounts of glucosamine and glucuronic acid as known for other heparin preparations. In protamine neutralization test the new heparin was 1.32 times more active than commercial heparin. The amidolytic method using chromogenic substrates S-2222 for factor Xa and S-2238 for thrombin demonstrated that the new heparin was also at least two times more effective than commercial heparin in increasing the rate of inactivation of these serin proteases through antithrombin III.

Characterization of a new potent heparin. 5th Communication: Further characterization by affinity chromatography on antithrombin-sepharose and its comparison with a commercially available heparin.

A highly potent heparin (Heparin Schering) was characterized in comparison to commercial heparin by affinity chromatography on antithrombin-III-sepharose. Heparin Schering consists of 83% high affinity and 17% low affinity fractions, whereas commercial heparin contained only 40% high affinity heparin and 60% low affinity heparin. The specific biological activities per microgram uronic acid and elution pattern on antithrombin-sepharose of high affinity heparin fraction were similar for both heparin preparations. It is concluded that the 2 times more potent anticoagulant activity of heparin Schering is mainly due to the enrichment of high affinity fraction (83%), which is also present in commercial heparin but at a remarkably lower percentage (40%).

Characterization of a new potent heparin. 3rd Communication: Determinations of anticoagulant activity of a new potent heparin preparation by thrombelastography in vitro using citrated dog and human blood.

The anticoagulant effect of a new potent heparin preparation was compared with a commercially available heparin by thrombelastography in vitro using citrated dog and human blood. The relative potency of the new heparin was found to be 1.45 to 1.77 with dog citrated blood and 1.77 to 2.41 with human citrated blood on the basis of different thrombelastographic parameters.