Effect of graded doses of naloxone on renal function in canine hemorrhagic shock. All the parameters of renal function (inulin clearance, para amino hippuric acid clearance and urine flow) which were depressed during experimentally induced hemorrhagic shock in dogs improved significantly in addition to improvement in mean arterial pressure (MAP) after bolus administration (iv) of 1 or 2 mg/kg naloxone. A smaller dose (0.5 mg/kg) of naloxone, however, did not improve the renal function. Even renal arterial injection of the same dose of naloxone showed no improvement in the renal function. In both these cases the improvement in the MAP was significantly less as compared to other groups of animals which received 1 or 2 mg/kg naloxone. It may be concluded that (a) naloxone at doses of 1 or 2 mg/kg improved the renal function by improving MAP and (b) naloxone has no direct action on renal vasculature.


Neurophysiological alterations following fluvalinate administration in mice. Median lethal dose (LD50) of fluvalinate (Marvik 25EC) was 105 (94.6-116.5 mg/kg, ip) in albino mice. Gross observable signs were dose dependent and indicative of central and peripheral nervous system stimulation. Fluvalinate, at 10.5 and 21.0 mg/kg, ip doses in mice, facilitated maximal electroshock seizures, reduced reaction time in analgesic test and enhanced duration of ether anaesthesia. Acute and subacute (7 days) treatment at lower and higher doses enhanced pentobarbitone sleeping time. Acute and subacute treatment (7 days) with phenobarbitone (50 mg/kg, ip) prior to fluvalinate enhanced toxicity of fluvalinate.


Effect of malathion on antioxidant defence system in human fetus--an in vitro study. Malathion under in vitro condition even at lower concentration (250 ppm) altered the level of enzymes associated with glutathione cycle and antioxidant defence system in human fetal brain and liver. Such changes involved alterations in glutathione status and extent of lipid peroxidation. The inhibitory effect of malathion was dose dependent in case of human fetal brain and was more vulnerable than fetal liver. This alteration (inhibition or activation) was maximum in case of tissues from fetuses of early period of development, suggesting greater susceptibility of human fetus towards this organophosphorus insecticide.


Effect of antioxidants on adenosine triphosphatase activity of brain and heart in male mice during ageing. The present study deals with the effect of butylated hydroxy anisole, propyl gallate and alpha-tocopherol on ATPase activity of brain and heart in ageing male mice. The activity of both the tissues significantly increased during pre-reproductive and reproductive phases. However, during the post-reproductive phase, ATPase activity of heart increased while that of the brain remained unaltered. The enhanced activity could be attributed to interference by the antioxidants in cellular metabolism and/or their free radical scavenging effect.


Characterization of a continuous lymphocyte cell line derived from BALB/c mice inoculated with a recombinant Moloney murine leukemia virus-TB. Neonatal BALB/c mice were inoculated (ip) with a recombinant Moloney murine leukemia virus-TB. Majority of the inoculated mice developed lymphoma within 5-7 months post infection. The cells from splenic lymphomas were cultured and 3 continuous cell lines (GP1, GP2 and GP3) developed. GP1 was single cell cloned and characterized. Based on Thy 1.2 (98.4%) phenotypic marker, the cell line was categorized as T cell line. The percent positivity for different cell surface markers on analysis with FACS was 98.4, 4.8, 5.5, 2.2, 1.8, 1.2 and 9.5 for Thy 1.2, mu, L3T4, Lyt2, Ia, IL2R and PNA receptor, respectively. A total of 16.5% GP1 cells was also positive for Moloney murine leukemia virus envelope protein (gp 70). Incomplete retrovirus like particles were demonstrated in the cytoplasm of GP1 cells by electron microscopy. The cell line on inoculation(ip) in neonatal BALB/c mice produced lymphomic lesions in almost all the vital organs of the mice.


Antigenic analysis of purified surface antigens of filarial nematode Setaria digitata. The surface antigens of S. digitata were isolated by treatment with Triton X-100. In non SDS-PAGE the surface antigen preparation resolved into more than 6 protein bands. Electroelution of gel slices corresponding to the protein bands with relative mobilities 0.09, 0.32, 0.41, 0.53, 0.61 and 0.76 gave 6 purified surface antigen fractions (SAF). Analysis of SAFs by SDS-PAGE showed that the proteins with molecular weights 17, 29 and 36 KD were the three major polypeptides and different combination of these gave rise to the 6 native surface proteins. The 29 KD protein existed as a monomer and as cross-linked with the 17 and 36 KD proteins. All surface antigen fractions showed antigenicity, where as 29 KD protein remained as a high avidity surface antigen.


Diosgenin--a growth stimulator of mammary gland of ovariectomized mouse. Estrogenic action of diosgenin on the mammary epithelium of ovariectomized (OVX) mouse has been reported. Diosgenin when administered (sc) at the dose levels of 20 and 40 mg/kg body weight for a period of 15 days stimulated the growth of mammary epithelium. This was indicated by the increase in DNA content, increase in number of ducts and appearance of terminal endbuds. There was a significant increase in the mammary development scores in the presence of diosgenin. Concomitant treatment of estrogen and diosgenin showed augmentation of estrogenic effect of diosgenin especially at the higher dose level (40 mg/kg body wt). Diosgenin showed a lack of progesterogenic action as was apparent from the absence of alveolar development even in the presence of exogenous estrogen.


Lytic effect of Bacillus subtilis elastase on gram-positive and negative bacteria. Elastase of B. subtilis 6a caused lysis of freshly grown cells of Gram-negative (Proteus vulgaris, Klebsiella pneumoniae, Salmonella typhi and Pseudomonas aeruginosa and Gram-positive (B. subtilis) bacteria. Heat killed and lyophilised Gram-positive and negative bacteria showed higher sensitivity to elastase. Both Gram-negative and Gram-positive bacteria were lysed maximally by elastase at pH 8.0. At this pH, activity of elastase was maximum in Tris-HCl and glycine-NaOH buffers followed by Tris-maleate and cacodylate buffers.


Modulatory effects of metanil yellow on immunity in rodents. Pathomorphological and immunological studies were carried out on rodents following oral administration of 0, 0.1, 0.25 and 0.5% (w/w) metanil yellow, mixed in diet, for 30 days. No significant change in hematologic parameters and histologic architecture of liver, kidney, mesenteric lymph node, thymus and urinary bladder was observed except for mild desquamation of intestinal villi and moderate changes in Peyer's patches of small intestine with higher doses. Among immunological parameters, significant enhancement in the primary humoral immune response (anti-SRBC IgM plaque forming cells of spleen) was observed with the lowest dose of metanil yellow while higher doses produced opposing effects. An elevated cutaneous delayed type hypersensitivity (DTH) reaction to SRBC was seen in 0.1% metanil yellow treated animals but higher doses did not influence the reaction. The treatment also caused changes in functional capabilities of macrophages. Although these immune alterations could hardly influence the local immunity of gut, as measured by the capacity of animals to cause rejection of Nippostrongylus brasiliensis parasite, the potential to modulate the immunity in general by metanil yellow however assumes considerable biological significance.


Toxic effects of different concentrations of dimethoate on lysosomal enzymes of female albino rats. The effect of three different concentrations of dimethoate on the activity of certain lysosomal enzymes, viz. beta-glucuronidase, beta-N-acetylglucosaminidase, cathepsin B and cathepsin D in serum, skin, liver, kidney and spleen and the stability of liver and kidney lysosomes was studied in female albino rats. The activity of beta-glucuronidase, beta-N-acetylglucosaminidase, cathepsin D was found to increase in serum and tissues in higher concentration (2.25 mg/100 g body weight) of dimethoate treated rats. A significant increase in the rate of release of beta-glucuronidase was found in the liver and kidney of higher concentration of dimethoate treated rats compared to controls. The results demonstrate that the activity of lysosomal enzymes increased in higher concentration of dimethoate treated rats than the lower concentration (0.56 mg/100 g body weight) of dimethoate treated rats.


Effect of styrene on testicular enzymes of growing rat. Effect of styrene (100 or 200 mg/kg body wt/day) for 60 days was observed on testicular enzymes of postnatally maturing rats. A significant decrease in epididymal spermatozoa count was observed only at 200 mg/kg body weight dose. Activities of testicular sorbitol dehydrogenase and acid phosphatase decreased while activities of lactate dehydrogenase, beta-glucuronidase, glucose-6-phosphate dehydrogenase, and gamma-glutamyl transpeptidase significantly increased only in animals exposed to styrene at a dose of 200 mg/kg body weight. The results suggest that exposure to high dose of styrene during developmental period alters the activities of enzymes associated with specific cell type of testis.


Trichloroethylene toxicity in mice: a biochemical, hematological and pathological assessment. Oral administration of trichloroethylene (TCE; 0, 500, 1000 and 2000 mg/kg/day) to male mice once daily, 5 days a week for a period of 28 days, caused a significant increase in liver weight, degeneration/necrosis of hepatocytes and characteristics proliferation of endothelial cells of hepatic sinusoids. Increase in kidney weight, glomerular nephrosis, degeneration/desquamation of tubular epithelium and characteristic amyloid deposition in glomeruli were observed only in the group of mice treated with 2000 mg/kg TCE. These changes occurred concurrently with a significant increase in total protein and free sulphydryl contents, elevated activities of acid phosphatase and catalase and decreased activity of delta-aminolevulinic acid dehydratase (delta-ALAD) indicating the sensitivity of liver and kidney as target tissues in TCE-toxicity. Hematological studies showed a significant increase in RBC counts and a reduction in WBC counts without any statistically significant change in the hemoglobin, urea nitrogen, creatinine and uric acid levels in the blood of TCE-exposed mice. A dose-related increase in cell density and acid phosphatase activity with a parallel significant decrease in the activity of delta-ALAD were observed in the bone marrow, which appear to be responsible for hematological alterations in TCE-exposed mice. The results suggest that early metabolic, pathological and hematological perturbations following a short-term exposure of TCE in mice, can provide the basis for its documented potential for chronic effects like blood dyscrasia and cancer.


Hepatic mixed function oxidase system and effect of phenobarbital, 3-methylcholanthrene and 1, 1, 1-trichloro-2, 2-bis(p-chlorophenyl)ethane in developing chickens. Contents of hepatic microsomal protein, aminopyrine N-demethylase, acetanilide hydroxylase, aniline hydroxylase, hydrogen peroxide formation, cytochrome-c-reductase, cytochrome b5 and cytochrome P-450 were examined in control, phenobarbital (PB), 3-methylcholanthrene (3-MC) and 1, 1, 1-trichloro-2, 2-bis(p-chlorophenyl)ethane (DDT) treated group of 1-28 days old chickens. Increase in aminopyrine N-demethylase, acetanilide hydroxylase, aniline hydroxylase, cytochrome-c-reductase, cytochrome b5 and cytochrome P-450 was noticed at all stages of development during administration of PB and 3-MC. But these enzyme activities were not always paralleled by increase in age. Aminopyrine N-demethylase was increased in early stages only during DDT administration, which indicates that the form of cytochrome P-450, responsible for aminopyrine N-demethylation is present in early stages only. However, acetanilide hydroxylase was decreased in all stages of development, in postnatal development the basal activities of the enzymes for various substrates do not exhibit identical pattern, the degree of inducibility by inducers varied in relation to age of animal. Hydrogen peroxide formation increased in all stages of developing chickens due to the administration of PB and DDT. It however decreased due to 3-MC administration which may be due to induction of high spin cytochrome P-450.


Acidic and basic protein contents in Gastrocnemii and pectoralis muscles of chick under denervation and work-induced stress conditions. Biochemical estimation of acidic and basic proteins of chick gastrocnemii (G. externus, G. medius and G. internus) and pectoralis muscles has been done under normal, denervated and work stress conditions from 1-56 days of postnatal growth. The reciprocal relationship of the two protein groups is clearly established. It is evident that muscle denervation acts as a stimulant for proteosynthetic activities and probably may also be an inhibitory factor for protein degradative reactions. During work overload stress, the rapid growth of muscles has been related to high rate of contractile activity.


Hypolipidemic action of taurine in rats. The effect of taurine on the serum and liver cholesterol and triglyceride levels was studied in rats fed cholesterol plus cholic acid. Four groups of 4 weeks old rats were fed control diet, hypercholesterolemic diet (HCD), HCD + 1% taurine or HCD + 2% taurine for 8 weeks. Addition of taurine in HCD diet showed a significant reduction not only in serum total cholesterol and triglyceride levels but also in liver total cholesterol, lipid and triglyceride contents compared to the animals fed HCD alone. Histological examination of organs of these animals showed severe fatty vacuolation in livers and signet ring type vacuolation in kidneys of rats fed HCD. Taurine showed ameliorating effect on these abnormalities. The animals fed taurine in HCD also showed increased bile and sterol excretion in faeces compared to rats fed HCD alone. Taurine showed significant hypocholesterolemia in rats probably by enhancing the catabolism of cholesterol and reducing the absorption of dietary cholesterol.


Effect of some natural pesticides on entomogenous muscardine fungi. Five commercial preparations of natural pesticides were tested for in vitro compatibility with muscardine fungi, Beauveria brongniartii and Metarhizium anisopliae. Neemark (azadirachtin) was found compatible with both the fungi. Phytoallexin, the natural fungicide, significantly inhibited the growth of both the fungi, while other natural pesticides showed moderate to severe inhibition.


Neurodepressive action of a piscicidal glycoside of plant, Aesculus indica (Colebr.) in fish. Sublethal concentration (2.6 mg/l) of a triterpene based piscicidal glycoside of A. indica damaged the neurons, fibre tracts and central correlation sites for gustatory, tactile and visceral sensory impulses in medulla oblongata of fish after prolonged poisoning. The fear, sinking to bottom, lack of schooling and non discriminatory pattern in treated fish were because of neurodepression. The jerky movement, ventilatory inefficiency, swallowing air bubbles and light pinkness of gills were due to the degradation of respiratory centres in the vagal lobes of toxified fish.


Effect of banana stem juice on biochemical changes in liver of normal and hyperoxaluric rats. Influence of stem extract of banana (family Musaceae), was studied on glycolic acid oxidase (GAO) and lactate dehydrogenase enzymes, calcium, phosphorus, oxalate and glycolic acid in liver tissues of sodium glycolate-induced hyperoxaluric rats. Activity of GAO was significantly lowered in the extract-treated rats compared to that of the glycolate-fed rats. LDH increased significantly in glycolate administered rats when compared with the extract-treated rats. The levels of calcium, phosphorus, oxalate and glycolic acid during hyperoxaluric state showed remarkable alterations in liver tissue.


Effect of dietary salt (NaCl) supplementation on urinary profile of guinea pigs (Cavia porcellus). Sodium chloride supplementation (120 mg/kg of body weight/day) for 12 days increased the urinary excretion of calcium from 91.6 +/- 9.0 to 159.4 +/- 16.0 mumol/day and of sulphate from 266.8 +/- 24.5 to 1176.9 +/- 87.2 mumol/day in guinea pigs. The stone risk due to increased urinary calcium excretion could possibly be counterbalanced by increasing urinary sulphate excretion. High salt intake, thus, could not increase the risk of stone formation.


A comparison of vessel sensitivity and vessel constant in oxygen uptake determination with pure oxygen and air as gaseous media. Air proved to be a superior gaseous medium to pure oxygen used for oxygen uptake study in the Warburg apparatus Recording of volume change (vessel sensitivity method) was the only correct method of recording the oxygen uptake. It is concluded that the volume change method should be followed for recording oxygen uptake by Warburg apparatus and pure oxygen should not be used as a gaseous medium.


Replicative activity of X-chromosome and autosomes of Drosophila melanogaster in autosomal hyperploids and the problem of dosage compensation. Replicative behaviour of two hyperploid autosomal arms (2L and 3L) of D. melanogaster has been analysed by 3H-thymidine autoradiography. Results reveal that hyperploid autosomal arms (2L-trisomy or 3L-trisomy) replicate synchronously with other disomic non-homologous chromosome arms i.e. there is no asynchrony in the initial mid or late phase of replication patterns between the trisomic 2L or trisomic 3L and disomic arms, suggesting that the extra dose of an autosomal arm can not alter the inherent pattern of replication of that arm. Results further reveal that 2L-trisomy or 3L-trisomy does not impart any influence on X-chromosomal replication in either sex. It is suggested from these results that change in the genomic dose of autosome does not play any role in modulating the replicative organization of the autosomes, 2L and 3L. Thus, although a regulatory mechanism of autosomal dosage compensation does exist for Drosophila, the hierarchy of genetic programming of regulation for X-chromosomal and autosomal dosage compensation might be different. Neither hypertranscriptive activity nor faster replication pattern of the male X-chromosome is influenced by 2L- or 3L-trisomy.


Differential response of testis and serum gonadotropins to testosterone in rats treated with a gonadotropin releasing hormone antagonist or estradiol 17 beta. Adult rats treated with a GnRH antagonist (Ac D2Nal1, D4Cl Phe2, DTrp3, DArg6, DAla10 GnRH; code: 103-289-10, National Institutes of Health, USA) for 5 weeks (250 micrograms/kg b.w) showed multiple degrees of impairment and atrophy of the genital organs concomitant with decreased serum levels of testosterone, LH and FSH. Inhibition of spermatogenesis was characterized by germ cell degeneration and overall decline in different cell numbers and in particular, spermatids of any kind were completely absent. Testosterone supplementation (60 micrograms/rat/day, sc) to GnRH antagonist treated rats, for the same period, significantly elevated the weights of the sex organs, and the serum levels of hormones. Spermatogenesis was improved both qualitatively and quantitatively; albeit failed to be restored back to control levels. Treatment with estradiol 17 beta (1 microgram/rat/day) for 5 weeks had insignificant effect on spermatogenesis but the weights of the genital organs (seminal vesicles by 19% and ventral prostate by 40%) and the levels of serum hormones (LH by 24%, FSH 22% and testosterone by 25%) were otherwise reduced. Administration of testosterone either alone or in combination with estradiol 17 beta had only a marginal effect on spermatogenesis or on other reproductive parameters. The results indicate a positive shift in the response of the testis and serum levels of gonadotropins to testosterone supplementation in rats treated with either GnRH antagonist or estradiol 17 beta.


Sperm production and fertility of bonnet monkeys (Macaca radiata) following immunization with ovine follicle stimulating hormone. Adult male bonnet monkeys were rendered oligospermic but not azoospermic following active immunization with ovine follicle stimulating hormone. The percentage of sperms in the semen having good motility was reduced with a concomitant increase in the sperm ATPase activity. Eight out of 10 immunized monkeys failed to impregnate females of proven fertility after mating for consecutive three cycles while the remaining two impregnated the cohabitated females during the third cycle at a time when the antibody titer was reduced. Active immunization with ovine follicle stimulating hormone may not produce complete azoospermia but renders adult male monkeys infertile provided sufficient antibody titer is maintained.


Neuropharmacology of amide derivatives of P-GABA. Three lipophilic amide derivatives of phthaloyl-GABA (P-GABA), namely gamma-phthalimido N-amyl butyramide (PGA), gamma-phthalimido-N-hexylbutyramide (PGH) and gamma-phthalimido N-phenylbutyramide (PGP), were synthesized and evaluated for their hypnotic and anticonvulsant activities in mice. Both PGA and PGH showed moderate hypnotic activity but PGP had no such action. Picrotoxin (0.08 mg/kg) a non-specific GABA antagonist completely abolished the hypnotic action of PGA in subconvulsive doses. Bicuculline (0.04 mg/kg) a GABAA antagonist, 3-mercaptopropionic acid (6 mg/kg) a GAD inhibitor at subconvulsive doses failed to neutralise the hypnotic action of PGA. On the other hand, PGA showed significant protection only against picrotoxin-induced convulsions, but was inactive against other convulsants tested. PGP which has no hypnotic activity, and has a mild anticonvulsant action in all the models except picrotoxin. A definite correlation was observed between the brain GABA and the hypnotic activity of PGA. However the present data indicate that the hypnotic and anticonvulsant activities are mediated probably through different brain GABA-ergic mechanisms.


Evaluation of some newer non-steroidal anti-inflammatory indan-1-acids in various biological systems. In line of the effort towards development of some newer indanyl non-steroidal anti-inflammatory agents and providing comprehensive SAR among this class of compounds some significantly active derivatives with low ulcerogenic potential were identified. Dealing with various long chain and branched chain compounds among this series, 3-(5, 6-dimethoxy indan-1-yl) propionic acid, 2-(5, 6-dimethoxy indan-1-yl) propionic acid and 3-(6-methoxy indan-1-yl) propionic acid were observed to have encouraging biological activity. Screening in various animal models of inflammation suggests their longer duration of action and lower ulcerogenic liability.


Mechanism of anti-stress activity of Ocimum sanctum Linn, eugenol and Tinospora malabarica in experimental animals. Effects of restraint stress (RS) and its modulation by O. sanctum (Os), eugenol and T. malabarica (Tm) were evaluated on some biochemical and biophysical parameters in rats. RS induced elevations in blood glucose and urea levels, were unaffected by either Os, eugenol or Tm pretreatment. However, both Os and eugenol lowered RS-induced cholesterol levels. RS also caused a generalized increase in enzyme activity and Os, eugenol or Tm effectively lowered the RS-induced elevations in lactate dehydrogenase (LDH) and alkaline phosphatase. RS also induced (a) increased membrane protein clusterization, (b) increased membrane fluidity and (c) reduced membrane thickness--in RBC membrane, whereas, the effects on the synaptosomal membrane were less marked. The RS-induced changes in RBC membrane dynamics were attenuated/reversed by Os, eugenol or Tm, in a differential manner. These biochemical and membrane changes during Rs and their modulation by the adaptogens are discussed in light of the possible mechanisms of action of these agents, during such aversive stimuli.


Effect of cadmium on tissue glutathione and glutathione peroxidase in rats: influence of selenium supplementation. Administration of cadmium (2.5 mg/kg, sc on alternate days for 3 weeks) to male albino rats led to significant accumulation of cadmium and metallothionein in the liver and kidneys. The activity of glutathione peroxidase was significantly decreased whereas, the concentration of glutathione was increased in these organs. Glycine-l-14C incorporation studies showed enhanced synthesis of glutathione in kidney but not in the liver. Selenium supplementation (1 mg/kg/day orally) failed to prevent these cadmium-induced changes, although it resulted in very high accumulation of selenium in these organs indicating the formation of cadmium-selenium complex.


Experimental pathogenicity of Mollicutes of bovine udder origin in hamster tracheal ring organ culture. Hamster tracheal-ring organ culture was employed to examine pathogenic effects of 8 isolates of Mollicutes of bovine udder origin. The tested Mollicutes could be categorized into two groups: (i) Mycoplasma F-38, M. mycoides var. capri, M. bovigenitalium mixed with M. bovirhinis, and M. bovigenitalium mixed with M. bovirhinis and Mycoplasma F-38 produced significant ciliostatic effect and infiltration of neutrophils and lymphocytes in lamina propria/subepithelium, hyperplasia and desquamation of epithelial lining cells and loss of cilia; and (ii) A. laidlawii, A. axanthum, an unidentified Acholeplasma and a mixed isolate of M. bovis, M. bovigenitalium, Mycoplasma F-38 and A. laidlawii showed insignificant ciliostatic effects and produced mild histopathological lesions. This correlates with the disease causing potentials of the strains.


Cardioprotective effect of cromakalim (potassium channel opener) in isoproterenol induced myocardial infarction in rats. Animals pretreated with cromakalim (1 mg/kg,po) along with isoproterenol (85 mg/kg,sc) showed less myocardial degenerative changes on histopathological examinations when compared with those treated with isoproterenol alone. Cromakalim's beneficial effects on myocardium were in dose-dependent manner. Administration of cromakalim (po) lowered significantly the serum LDH and SGOT and depleted intracytoplasmic glycogen as demonstrated by periodic schiff staining procedure. Increase in blood clotting time was highly significant (P less than 0.001). The results suggest cardioprotective effect of cromakalim in isoproterenol induced myocardial infarction.


Effect of doxorubicin on heart mitochondrial enzymes in rats: a protective role for alpha-tocopherol. Effect of doxorubicin on heart mitochondrial enzymes was studied in rats with or without the administration of alpha-tocopherol. Rats were treated with doxorubicin 2.5 mg/kg, ip body wt once a week for 8 weeks. Alpha-tocopherol was co-administered orally for 2 months (400 mg/kg body wt daily). TCA cycle enzyme, NADH-dehydrogenase, cytochrome-C-oxidase and Na+,K(+)-ATPase activities were found to be decreased in doxorubicin treatment. A significant decrease in protease activity was observed with a concomitant increase in mitochondrial protein level. Mitochondrial lipid peroxide level was found to be increased with a decrease in thiol content. Alpha-tocopherol co-administration was found to maintain the mitochondrial enzyme activities as well as the thiol content. The results are discussed with reference to the antioxidant nature of alpha-tocopherol.


Effect of five-membered heterocycles against Leishmania donovani infection. Impact of change of heteroatom in pentavalent heterocycles, viz., pyrroles, isoxazoles, imidazoles and crotonates on the profile of antileishmanial activity against amastigotes of L. donovani using in vivo test system and macrophage-amastigote culture system has been studied. Sixty-three compounds were tested. Nine imidazoles showed marginal activity in vivo, whereas 3 out of 10 compounds of isoxazolone series and 2 out of 4 substituted aminocrotonates exhibited antileishmanial activity. Of the 30 substituted pyrroles, except 8 all showed antileishmanial activity in vivo on day 7 post treatment.


Autoimmunity in amoebiasis.

Autoimmunity in amoebiasis. Amoebic liver abscess (ALA) and symptomatic intestinal amoebiasis cases were assessed by indirect haemagglutination assay for auto-reactive IgG and IgA class of antibodies in response to healthy human serum IgG and IgA. The present results indicated the presence of autoreactive IgG and IgA class of antibodies in ALA and intestinal amoebiasis respectively.


Bromocriptine-induced implantation failure in mice: failure of the stud male to protect pregnancy. The presence of the stud male failed to prevent implantation failure in newly inseminated females induced by administration of bromocriptine. This is in contrast with the ability of the stud male to prevent implantation failure in females induced by alien male exposure, and nutritional stress. Since bromocriptine is a potent inhibitor of hypophysial prolactin release by virtue of its stimulatory effect on hypothalamic dopaminergic activity, the results suggest that the stud male-originating luteotrophic stimulus is incapable of overriding the dopaminergic activity in bromocriptine-treated females.


Lung injury by amiodarone, an antiarrhythmic drug, in male rats. Administration of single dose (175 mg/kg body wt) of amiodarone dissolved in water through gavage for 3 weeks damaged the lung and changed the content of lung lavage. Activities of bronchoalveolar lavage (BAL) angiotensin converting enzyme (ACE) and lactate dehydrogenase (LDH) increased significantly. Also, the protein and lactate content of the lavage fluid increased significantly over the control. The drug also produced marked changes in morphology of the lung of experimental animals.


Cervix tumour cell line established from tumour after long term passages as heterotransplant in nude mice. One human cancer of the uterine cervix xenograft was established in tissue culture only after repeated passages in nude mice suggesting that with the repeated passages in nude mice, tumour cells acquire some properties which allow them to grow in vitro. Attempts to establish cell line tumours from earlier passages were not successful. The established cell line is tumourigenic. On inoculation of cultured cells in nude mice tumour take was found to be 100%. Karyotypic analysis revealed human origin.


Tissue changes induced by enterotoxic preparations of Salmonella weltevreden in ligated rabbit gut segments. Culture filtrates and cell lysates of two strains of S. weltevreden which caused dilation of ligated rabbit gut segments (characteristic associated with the enterotoxic activity) induced mild to severe architectural changes in the test segments of intestine. The dilated segments contained thick, bloody and mucoid exudates. The results suggested that besides invasiveness and enterotoxigenicity, S. weltevreden possibly possessed factor (s) that damaged intestinal tissue and played part in the pathogenesis of Salmonella gastroenteritis.


Cloning and nucleotide sequencing of sheep growth hormone cDNA. cDNA was prepared from the mRNA isolated from sheep anterior pituitary glands. On cloning cDNA in E. coli, a clone coding full sequence of sheep pre-growth hormone was determined. The sequence for the sheep growth hormone (GH) is in agreement with the amino acid sequence of the protein determined previously except for the asparagine residue at position 99 rather than aspartic acid and the arginine residue at position 146 in place of threonine. The cDNA sequence presented is also in accordance with the genomic sequence for the sheep GH gene that has been reported.


Effect of storage duration on cell cycle kinetics and yield of chromosomal aberrations in PHA-responsive human peripheral blood lymphocytes. Phytohaemagglutinin (PHA)-responsive lymphocytes from human peripheral blood samples, either irradiated or un-irradiated, showed increased frequency of first division metaphase cells (detected by fluorescence plus Giemsa (FPG) staining) as a function of duration of storage. Irradiated and subsequently stored samples showed small but significant increase for the yield of dicentrics. The yield of aberrant metaphases and deletions (excess acentrics) remained unchanged. Increasing Bromodeoxyuridine (BrdU) concentrations slowed down the cell cycle progression but did not influence the yield of aberrations including that of dicentrics.


Effects of hCG and PMSG on responsiveness of ovary during period of delayed ovulation in an Indian vespertilionid bat, Scotophilus heathi. Both hCG and PMSG treatments given either individually or sequentially between September and early February failed to induce ovulation in S. heathi, though they produced high degree of ovarian stimulation. The treatments induced ovarian enlargement, intense hyperaemia, marked changes in interstitial cells (ICs) and development of several antral follicles and varying degrees of abnormality in oocyte of most of the antral follicles. The percentage of abnormal oocytes in the ovary following hCG or PMSG treatment was dose-dependent.


Influence of different photoperiods on development of gonad, cloacal gland and circulating thyroid hormones in male Japanese quail Coturnix coturnix japonica. One day old chicks of Japanese quail were exposed to different photoperiods (LD, 8:16, 13.5:10.5, 16:8 and LL) and observations (testes weight, cloacal gland size, body weight and circulating thyroxine and triiodothyronine) were taken at the age of 3, 5, 7, 9 and 16 weeks. Results indicate that immediate reproductive development occurred in birds exposed to long photoperiods (greater than 12 hr). Growth under LD 8:16, was not apparent till 7th week and by 16 weeks, degree of gonadal development was similar in all the birds, irrespective of photoperiodic treatment. Whereas body weight of the intermediate and long day (LD 13.5:10.5, 16:8 and LL) treated birds increased upto 5th week and remained constant thereafter. But the chicks maintained under short day length (LD 8:16), showed spontaneous increase till the end of the study and birds were much heavier compared to all other groups. Plasma T4 concentration increased with increasing age till 9th week and remained unaltered thereafter. On the other hand T3 level did not change till 7th week followed by a decline. It is suggested that the initiation and degree of gonadal growth in quail depends on the availability of daily photoperiod, until the achievement of full breeding condition. Peak level of T4 observed in 9 week old birds may be involved in the development of photorefractoriness at that age.


Effect of divalent metal ions on soluble and membrane-bound alkaline phosphatase activity in suckling rat intestine. EDTA treatment of intestinal brush border membranes (BBM) and epithelial cell supernatant completely inhibited alkaline phosphatase (AP) activity in suckling rat intestine. AP activity was fully regained upon dialysis of the preparations against Zn2+ and to a lesser extent against Co2+, Ca2+ and Mn2+ ions. Other metal ions (Cd2+ and Mg2+) tested were essentially ineffective in restoring the enzyme activity. Considerable differences were observed in kinetic characteristics of the membrane-bound and soluble AP activities in response to various metal ions. There were apparent differences in Km, Vmax, energy of activation (Ea) and thermal stability of the soluble and membrane-bound AP activities, after metal ion substitutions. Nearly 35% AP activity was solubilized on sodium dodecyl sulphate treatment of brush borders (membrane protein: detergent ratio 1:3; w/w). Dialysis of detergent solubilized BBM against different metal ions reconstituted AP activity in the particulate fraction: the order of effectiveness was Zn greater than Ca greater than Mn greater than Co. The kinetic properties of the reconstituted AP were essentially similar to the non-integrated enzyme activity in response to various divalent metal ions examined. But there were apparent differences in Km, Vmax, Ea and thermal stability of the reconstituted AP activity compared to native brush border enzyme. The results suggest the unique requirement of Zn ions for stability and catalytic activity of the soluble and membrane-bound AP activity in suckling rat intestine.


Induction of tumour necrosis factor alpha in experimental animals treated with hepatotoxicants. Administrations of hepatotoxicants namely carbon tetrachloride (CCl4:0.4 ml in 1.2 ml of liquid paraffin) and ANIT (1 ml of 1.5% solution in liquid paraffin) in Charles foster rats (force fed) and D-galactosamine (8 mg in water per swiss albino mouse, ip) induce the release of TNF-alpha in case of CCl4 and D-galactosamine. High TNF-alpha level was observed up to 48 hr in CCl4 and up to 24 hr in D-galactosamine treated animals. Elevated levels of biochemical like ALP and SGPT are also recorded. TNF-alpha level can be measured of tissue damage and prognosis in case of hepatitis.


Insecticide induced disruptions in functioning of developing brain of Rana cyanophlictis. Synthetic pyrethroid insecticide permethrin significantly decreased the levels of regulatory proteins (S-100 and calmodulin) in the developing CNS of tadpoles of R. cyanophlictis. Remarkable inhibition of enzymes acetylcholinesterase and choline acetylase and significant accumulation of neurotransmitter acetylcholine were observed in permethrin treated animals. Permethrin exposure significantly decreased the activity of phosphodiesterase. The results support molecular disruptions occurring due to permethrin induced toxicity. This in turn may bring about neuronal inefficiency in the treated tadpoles.


Effect of citrate feeding on free radical induced changes in experimental urolithiasis. Feeding calculi producing diet (CPD) to rats for 4 weeks produced calcium oxaltate stones. Supplementation of sodium citrate to CPD (c-CPD) prevented stone formation. Except oxalate, the excretion of calcium, phosphorus and magnesium was restored to normal in c-CPD fed rats. The CPD fed rats exhibited increase in glycolic acid oxidase (GAO) and lactate dehydrogenase (LDH) activities and only GAO activity was partially restored in c-CPD fed rats. Kidney sub-cellular fractions of calculi producing diet (CPD) fed rats showed increased susceptibility for lipid peroxidation in presence of promotors. Antioxidant enzyme activities of superoxide dismutase (SOD), catalase and glutathione peroxidase and antioxidant concentrations of reduced glutathione, total thiols, ascorbic acid and vitamin E were significantly decreased while the xanthine oxidase activity, and concentrations of hydroxyl radical, diene conjugates and hydroperoxides were significantly increased in CPD fed rats. The susceptibility to lipid peroxidation, activities of antioxidant enzymes, and the concentration of antioxidants were not normalized by feeding citrate.


Effect of picroliv on glutathione metabolism in liver and brain of Mastomys natalensis infected with Plasmodium berghei. Administration of picroliv, the active principle from Picrorhiza kurrooa, at a dose of 6 mg/kg, po for two weeks showed significant protection against changes in liver and brain glutathione metabolism of Plasmodium berghei infected Mastomys natalensis. The depletion of reduced glutathione level and inhibition of glutathione-S-transferase, glutathione reductase and glutathione peroxidase activities due to P. berghei infection were markedly recovered by picroliv. The increased levels of lipid peroxidation products in damaged tissues were also reduced along with the recovery of glutathione metabolism.


Effect of gossypol on tissue levels of glutathione and thiobarbituric acid reactive products in rats. Gossypol was administered in pubertal and adult rats and lipid peroxide formation and GSH levels were estimated in different tissues like liver, testis, heart and kidney. Gossypol caused low generation of lipid peroxides, measured as thiobarbituric acid reactive products (TBAR), without causing significant changes in tissue glutathione (GSH) levels. This effect was more pronounced in liver and testis as compared to other tissues. In vitro effect of gossypol to inhibit lipid peroxidation as observed in vivo suggested that binding of gossypol to plasma membranes may result in inhibition of lipid peroxide generation.


Elevation of C-reactive protein in serum of Channa punctatus as an indicator of water pollution. Effect of some pollutants like heavy metals, non-metals and pesticides on the circulating level of C-reactive protein (CRP) which is an acute phase plasma protein was studied in a freshwater murrel C. punctatus. Fish was exposed to nonlethal doses of these xenobiotics which were apparently safe. But the level of CRP detected by sensitive single radial immunodiffusion (SRID) technique showed that within 12 hr of exposure the nonlethal doses of xenobiotics could initiate the acute phase response in terms of elevated CRP titre. Heavy metals caused the acute phase within 24 hr, nonmetals and Metacid-50 within 48 hr exposure. The carbamate compound, carbaryl demonstrated a biphasic response to CRP level which may be correlated with the reversible type of anticholinesterase property of this compound while Metacid-50 is an irreversible type of anticholinesterase agent. The assessment of the CRP level in the serum of fish may be utilized as a primary bioindicator of a contaminated environment toxic enough to mount an acute phase response.


Hepatoprotective activity of Azadirachta indica leaves on paracetamol induced hepatic damage in rats. Effect of A. indica leaf extract on serum enzyme levels (glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, acid phosphatase and alkaline phosphatase) elevated by paracetamol in rats was studied with a view to observe any possible hepatoprotective effect of this plant. It was interesting to observe that serum enzyme levels were much elevated in paracetamol induced animals than in those receiving a combination of paracetamol and lead extract. It is stipulated that the extract treated group was protected from hepatic cell damage caused by paracetamol induction. The findings were further confirmed by histopathological study of liver.


Role of pure and biologically active amoebal RNA in assessment of lymphokines: a report of 55 ALA cases. Amoebic liver abscess cases (55) were assessed for release of lymphokines (LMIF) using pure and biologically active amoebal RNA of axenic Entamoeba histolytica (NIH: 200) obtained with cesium chloride centrifugation. Lymphokines released by T lymphocytes in response to both amoebal RNA and whole amoebic lysate (WAL) were tested by leukocyte migration inhibition test (LMIT) on blood samples from amoebic liver abscess cases. A significant increase was observed in the release of lymphokine and 100% positivity was observed with amoebal RNA compared to whole amoebic extract with a positivity of only 78%. The difference between means leukocyte migration inhibition of the above two with regards to release of lymphokine was highly significant (P less than 0.001). This shows that patients had high degree of leukocyte sensitization to amoebal RNA of E. histolytica compared to whole amoebic lysate. These findings suggest that the amoebal RNA plays an important role as a potent antigen in the elicitation of cell mediated immune responses in amoebic liver abscess cases.


Disposition of norepinephrine and epinephrine in jejunum of WLH chicken. Disposition of (-)-norepinephrine and (-)-epinephrine in jejunum of WLH chicken was studied using oil-immersion technique. The relative rate of different routes of disposition of catecholamines was in the following order: for (-)-NE COMT greater than or equal to MAO greater than or equal to U2 greater than U1, for (-)-Epi U2 greater than or equal to COMT greater than MAO greater than U1. The role of enzymatic degradation is almost equal to that of uptake processes for (-)-Epi, but it was greater for (-)-NE.


Relative ranking of culture media based on proliferation kinetics and spontaneous chromosomal aberrations in PHA-responsive human peripheral blood lymphocytes. Proliferation kinetics and spontaneous yield of chromosomal aberrations phytohemagglutinin (PHA)-responsive peripheral blood lymphocytes were studied from blood samples collected from 45 individuals in 4 different synthetic media. Except for a significant difference for Eagle's MEM and RPMI 1640, the other media did not show difference for the yield of chromatid or chromosome type of aberrations. Differences were however noticed in the proliferation kinetics (mitotic and proliferative rate indices) of cells among the media used. The study indicated that (i) the intrinsic properties of media which influence proliferation rate and yield of chromosomal aberrations are independent of each other as higher proportion of first division cells do not correspond with higher frequency of chromosomal aberrations, (ii) the amount of free-radical scavengers present in the medium, apart from the genetic make-up of the individuals, may contribute to the spontaneous yield of chromosomal aberrations and (iii) RPMI 1640 medium, which showed higher transformation and faster cycling rate for the lymphocytes, may be considered as medium of choice for analysing two main cytogenetic end-points, chromosomal aberrations and sister chromatid exchanges (SCEs).


Presence of RNA from yeast inhibits the photoreactivation of UV-irradiated DNA by Phr A photolyase from Escherichia coli. Photoreactivation of UV-irradiated DSNA with phr A photolyase from Escherichia coli was studied in the presence of yeast RNA. Mixing of RNA with UV-irradiated DNA before its treatment with photolyase inhibited the photoreactivation of DNA. Denatured (by sonication) RNA was found to be more effective in blocking photolyase action. Agarose gel electrophoresis experiments suggest that this inhibition of photoreactivation is due to interference in the binding of photolyase with UV-irradiated DNA by yeast RNA.


Cold abscess due to malignant pleural mesothelioma. A middle aged male presented to us with a cold abscess on the left side of chest which later proved to be a malignant pleural mesothelioma. The clinical, radiological and histopathological findings have been discussed.


Cockroach in right main bronchus--an unusual foreign body. Foreign body aspiration in children is not a uncommon clinical problem. Usually the foreign body is of vegetable origin. It is distinctly uncommon to find a living organism like cockroach, especially a large one as a foreign body in respiratory tract. Here we report one such rare experience.


Fracture of tracheostomy tubes--report of 3 cases. Fracture of the tubes appears to be caused by their prolonged use and defective fusion at the junction of cannula and phalange. We report here three cases of fractured polyvinyl chloride tracheostomy tube, an unusual complication of tracheostomy, presenting as foreign body in the trachea.


Evaluation of unguided transbronchial biopsy in the diagnosis of pulmonary disease--its safety and efficacy as an out-patient procedure. Sixty-five patients underwent unguided fiberoptic transbronchial biopsy on an out-patient basis. Satisfactory material for histopathological examination was obtained in 55 patients. A reasonably definite pathological diagnosis could be established in 51 patients. The commonest diagnosis arrived at was that of interstitial fibrosis. Complications were observed in 4 (6.2%) patients. Fiberoptic transbronchial biopsy is, therefore, a safe and efficacious out-patient technique for establishing a pathological diagnosis in patients with infiltrative, diffuse or localized pulmonary disease.


Farmer's lung disease in north-western India--a preliminary report. A study of farmer's lung (FL) disease was carried out in 197 subjects engaged in farming and having respiratory complaints of varying duration. It revealed that 13.2% of the subjects had precipitating antibodies against thermophilic actinomycetes, with Faenia rectivirgula (Micropolyspora faeni) alone accounting for 85% of the positive reactions. Precipitating antibodies against Thermoactinomyces vulgaris and T. thalpophilus were observed only in 1.5% and 0.5% of the subjects, respectively. Two subjects concomitantly demonstrated F. rectivirgula and T. vulgaris-specific serum precipitins. Sixty (30%) of the subjects related their respiratory symptoms to exposure to wheat straw/thresher's dust or other vegetable substrata in the working environment. Based upon a suggestive clinical history, roentgenography, pulmonary function studies and demonstration of serum precipitins against F. rectivirgula, FL was diagnosed in 4 subjects whose salient features are presented and discussed. To the best of our knowledge, this is the first authentic report on FL from India. A comprehensive epidemiological survey is indicated to determine the prevalence of FL in different geo-climatic regions of the country.


Inhibition of acetylcholinesterase of mice erythrocytes & synaptosomes by dimethylsulfoxide. In vitro inhibitory effect of dimethylsulfoxide (DMSO) on acetylcholinesterase of erythrocyte membranes and synaptosomes of mice was observed using acetylthiocholine as substrate. DMSO inhibited the enzyme from both sources and the effect was concentration dependent. It produced an inhibition of 26 to 28 per cent at a concentration of 0.13 mM and 92 to 95 per cent at a higher concentration of 1.91 mM. The Km of bound and solubilized enzyme of the synaptosomes was 0.2 and 0.15 mM while that of erythrocyte membranes 0.5 and 0.33 mM respectively. The Vmax was 0.4 and 0.4 (for synaptosomes) and 0.67 and 0.59 (for erythrocyte membranes) Mmoles/mg protein/min, respectively for bound and solubilized forms. The Ki of solubilized enzyme of synaptosomes was 0.11 mM. Kinetic studies showed that the inhibition was competitive in nature.


Intensive respiratory care service--our eight-year experience. Intensive respiratory care service has come a long way from the beginning in Scandinavia in early 1952. Intensive care service (IRCU) was started at our institute in 1983 as tetanus and respiratory care ward which has developed into Department of respiratory diseases over years. We are reporting our experience of 886 cases of acute respiratory failure (ARF) treated from Jan, 1983 upto April 1990.


Perfusion lung scan in tropical pulmonary eosinophilia. Perfusion lung scan using macroaggregate of human serum albumin tagged with I131 was done in 7 patients with tropical pulmonary eosinophilia (TPE). In the 3 untreated patients perfusion defect of varying degree could be noticed, whereas in 4 other patients with history of chronic illness and previous treatment with multiple courses of diethylcarbamazine the lung scan was normal. Possible pathophysiology in light of above findings in TPE is discussed.


Serum lipids & lipoprotein profiles of cigarette smokers & passive smokers. Serum lipids and lipoproteins of 50 active and passive smokers were compared with levels in 25 control subjects. Active smoking resulted in an increase in total cholesterol (Tc) and triglycerides (Tg) as compared to control group. The passive smokers also showed relatively higher levels but the effect was not significant. Active smoking raised the low density lipoprotein cholesterol (LDLc) and very low density lipoprotein cholesterol (VLDLc) levels whereas high density lipoprotein cholesterol (HDLc) content was lowered, thus resulting in decreased ratios of HDLc/Tc and HDLc/LDLc. The passive smokers also showed slightly higher levels of LDLc and VLDLc but lower levels of HDLc, and a lower HDLc/LDLc ratio. Our findings suggest that smoking alters the serum lipids and lipoproteins and these changes are related to the duration and amount of smoking.


Effect on curcumin on cholesterol gall-stone induction in mice. A study was carried out on the efficacy of curcumin in reducing the incidence of cholesterol gall-stones (CGS), induced by feeding a lithogenic diet in young male mice. Feeding a lithogenic diet supplemented with 0.5 per cent curcumin for 10 wk reduced the incidence of gall-stone formation to 26 per cent, as compared to 100 per cent incidence in the group fed with lithogenic diet alone. Biliary cholesterol concentration was also significantly reduced by curcumin feeding. The lithogenic index which was 1.09 in the cholesterol fed group was reduced to 0.43 in the 0.5 per cent curcumin supplemented group. Further, the cholesterol: phospholipid (C/PL) ratio of bile was also reduced significantly when 0.5 per cent curcumin supplemented diet was fed. A dose-response study with 0.2, 0.5 and 1.0 per cent curcumin supplemented lithogenic diets showed that 0.5 per cent curcumin was more effective than a diet with 0.2 or 1 per cent curcumin.


Effect of three antibacterial drugs in lowering blood & stool ammonia production in hepatic encephalopathy. Neomycin (700 mg/8 h), ampicillin (500/6 h) and metronidazole (400 mg/8 h), were compared for their effect, on oral administration for 4 days, in reducing blood ammonia in 27 patients with stable chronic liver disease. It was found that there was 38.2, 38.5 and 8.7 m mol/litre mean reduction in blood ammonia in the neomycin, ampicillin and metronidazole treated groups respectively. The difference in blood ammonia was statistically significant for both neomycin (P = 0.01) and ampicillin (P = 0.03) but there was no significant change after metronidazole treatment (P = 0.6). The total stool enzyme activity at optimum pH was maximally reduced by ampicillin and minimally with metronidazole. The reduction was noted to be 3.51 m mol/1 (P = 0.01), 3.87 m mol/1 (P = 0.08) and 2.8 m mol/1 (P = 0.02) of NH3/g dry weight of stool for neomycin, ampicillin and metronidazole respectively. The main bacterial gut enzymes responsible for ammonia production, urease and protease, were found to be very sensitive to stool pH. At pH 6 their activity was around 20 per cent of what was found in optimum pH of 7.4 and at pH 5 it is only about 8 per cent of optimum activity. None of the three antibacterial agents changed the stool pH significantly. It can be concluded that oral neomycin and ampicillin are superior to oral metronidazole in lowering blood ammonia.


Acute accidental exposure to chlorine fumes--a study of 82 cases. Eighty-two patients were hospitalized following an accidental exposure to chlorine. All patients presented with dyspnoea and cough. The other symptoms included irritation of throat (53.6%), irritation of eyes (42.3%), headache (29.2%), abdominal pain (26.8%), vomiting (24.3%) and giddiness (9.7%). All of them had bronchospasm and 5 (6%) had cyanosis at the onset. An x-ray of the chest revealed patchy infiltrates in 3 (3.85%) and hilar congestion in 2 (2.44%). Pulmonary function tests showed an obstructive pattern in 27.4%, restrictive in 3.25% and mixed in 53.2%. Pulmonary functions were normal in 16.1% of the patients. Bronchoscopy revealed tracheobronchial mucosal congestion in all cases, hemorrhagic spots in 35.7%, erosions and ulcers in 12.5%. All patients were treated with oxygen, aminophylline, hydrocortisone and antibiotics. Haematemesis (n = 1) and pulmonary oedema (n = 2) developed 12 hours after the admission. Two other patients developed pneumonia 48 hours later. All patients recovered satisfactorily. On follow-up 16 patients had no sequelae after one year. Pulmonary functions were normal in 5 patients after 3 years of follow-up.


Effect of growth on lung transfer factor and its components. Measurement of lung transfer factor for CO (TLCO) and its constituent components, viz. diffusion capacity across alveolar capillary membrane (Dm) and instant pulmonary capillary blood volume (Vc) were undertaken in 120 healthy non-smoker males by single breath technique. All the three parameters (TLCO, Dm, Vc) showed direct negative correlation with age. While TLCO and Dm showed a significant direct positive correlation with height, there was no correlation between VC and height. The degree of correlation increased when both age and height were used together than either of them alone for prediction of TLCO120. The prediction formulae (TLCO120 = 3.8 + 21 H (m)-0.308 A) using both age and height has regression value (R) of 0.6479 (p < 0.001).


Detection of gene deletion in patients of Duchenne muscular dystrophy/Becker muscular dystrophy using polymerase chain reaction. Polymerase chain reaction (PCR) was used to study the presence of gene deletion (the most prominent type of mutations) in some families afflicted by Duchenne muscular dystrophy/Becker muscular dystrophy (DMD/BMD). The results clearly demonstrate deletion in the central part of the DMD gene in two of the three families studied. This information can be useful for genetic counselling with particular reference to prenatal diagnosis and carrier analysis.


Lung cancer in India--a perspective.

Lung cancer in India--a perspective. The epidemiology, aetiopathogenesis, diagnostic methods and management of lung cancer in India have been reviewed based on the published data from this country. Over the years, the incidence has been increasing and the disease is being recognized more often. Smoking is the most important aetiological factor. Beedi smoking which is a common practice in this country is equally, if not more, harmful than cigarette smoking in causing lung cancer. Tremendous improvement has been made in various diagnostic techniques. Cytological methods, fiberoptic bronchoscopy and percutaneous lung aspiration are being used more frequently. At the treatment front, although radiotherapy and chemotherapy are being used, the results are not very encouraging. Except in few cases, surgery is usually not feasible because of advanced disease. Prevention of lung cancer by anti-smoking measures should be the main goal of solving the problem in this country.


Induction of sister chromatid exchanges in phenytoin treated & untreated patients with epilepsy. The mutagenic potential of phenytoin (PHT) was studied using the sister chromatid exchange (SCE) assay. Twenty nine PHT treated epileptics, 32 untreated and 32 normal healthy controls were analysed. Similar SCE frequencies were observed in untreated patients and patients on PHT monotherapy. Both groups had significantly increased SCE frequency as compared to controls. No positive correlation of SCE frequency with sex and duration of therapy was observed. The results of the present study suggest the role of the disease condition in inducing genetic damage as assessed by increased SCE frequencies.


Perinatal mortality in Ludhiana, Punjab--a seven year hospital study. Perinatal deaths occurring over a seven year period were studied in a teaching hospital in Punjab. Causes of death were analysed as per Wigglesworth's classification. We have further modified this by correlating different gestations and weight groups with causes of death. The perinatal mortality rate (PNMR) in the present study was 74/1000 and showed a downward trend secondary to a statistically significant fall of early neonatal mortality. There was a decline in PNMR among babies of 1500-2000 g. birth weight and 33-36 weeks gestation. Asphyxia and macerated still births were found to be the two main causes of death. Macerated still births were seen more commonly among babies of lower weight at all gestations. The PNMR of babies born to booked mothers was 22/1000 as compared to 152/1000 among unbooked mothers. It was concluded that to bring down the PNMR, economic development alone is not enough. Provision of adequate antenatal care to all mothers, health education and timely referral of high risk mothers is very essential.


Socio-biological factors in underfive deaths in a rural area. In 1985-86, 286 underfive deaths occurred among a population of 30,000 in a rural area of Haryana. Two hundred and eighty one were analysed for socio-biological factors related to under five mortality. Females had a higher mortality. About 2/3 of the deaths were in infants, and 90% in first 3 years. Most of the deaths (94%) occurred in the village itself, 58.4% did not seek any medical care during the terminal illness, 80-90% did not receive even a single dose of BCG, DPT or O.P.V., and 36.7% died in the first attack of illness. Though 68% had at least one episode earlier, 31.0% had been admitted in hospitals for an earlier episode of illness. In 42.8% of deaths, the birth order was IV or above. Deaths in socially and economically disadvantaged cases constituted 77.6%. The triad of diarrhea, ARI and malnutrition claimed 56% deaths. In 93% of the deaths, the mothers were illiterate and 96.4% were house wives. There was a sibling death earlier in the family in 78.3%, and 60.1% deaths were of those living in poor housing conditions. About 50% had radio for communication, 85.8% had bicycle for conveyance, and in 66.9% the family had piped water supply. All these findings have been discussed in the study.


Severity of rheumatic mitral regurgitation. Thirty patients below the age of 15 years (range 8-1/2 to 15 years, mean 11.8 years) have been studied for the severity of rheumatic mitral regurgitation. Moderately severe to severe pulmonary venous hypertension was found in 76.6% and pulmonary arterial hypertension in 60%. Left ventricular volumes could be calculated in 13 patients. The end-diastolic volume was elevated in 11 and the endsystolic volume in 12 cases. The regurgitant fraction, calculated in nine patients was 0.6 or more in seven cases. The clinical and hemodynamic severity of mitral regurgitation in children was identical to that seen in adults in the absence of active rheumatic carditis. Children with dominant rheumatic mitral regurgitation can develop congestive failure on the basis of valvar damage per se.


Infant feeding practices in rural Bangladesh. A longitudinal study was done on the infant feeding practices in a rural area. One hundred and ten infants were followed up from birth to 1 year of age by alternate day home visits, to inquire about the type of food, and frequency of consuming it. It was found that 100% mothers breast-fed their infants from birth to 1 year, almost every day. But, bottles containing various kinds of milk and starchy food were added to 60% of infants diets by 3 months, and 80% by 5 months of age. This additional food was given mostly in diluted form, which was more so in case of tinned milk. Family food such as rice and vegetables were given in 30% and 40% child days respectively from 6 months to 1 year. Rural people withhold protein food and fruits during infancy. It is concluded, that infant feeding practices in our population is improper and mothers should, therefore, be trained and motivated on weaning practices for timely and adequate supplementation to ameliorate the presently observed dietary deficiency and early malnutrition in rural Bangladesh.


Prevalence of iodine deficiency disorders in adolescents. In the present study the iodine status of 300 adolescent boys and girls was assessed by clinical examination and biochemical tests. The clinical examination revealed the total goitre rate (TGR) to be 65.2% among boys, and 69.6% among girls. The visible goitre rate (VGR) was 17.7% among boys and 21.1% among girls. Nutritional status of all adolescents was found to be poor as compared to their well-nourished counterparts. Using discriminant analysis it was found that age, height and weight of the adolescents were significantly related to goitre grade (p < 0.001), and they are important in prediction of goitre. Mild and moderate iodine deficiency were found to be prevalent among the adolescents. On the basis of urinary iodine/creatinine ratio, 38% of the adolescents were found to be suffering from mild iodine deficiency, i.e. average urinary iodine excretion between 50-100 mcg iodine/g creatinine. Moderate iodine deficiency (< 50 mcg iodine/g creatinine) was found to be prevalent among 12.4% of the adolescents. The results of this study indicate a high prevalence of mild and moderate IDD among the adolescents studied.


Electrical response audiometry (ERA) in infants. The electric response audiometry is a useful diagnostic technique for identifying hearing loss in infants. Among the various electrical responses, the BSER enables us to arrive at an accurate early clinical diagnosis of hearing loss in infants so that effective auditory input, which is the prime requisite for the conceptual foundation for the growth of communication, can be provided through appropriate intervention programmes. The electric response audiometry provides information regarding the physiologic state of the peripheral organs and auditory pathways but however, cannot ascertain whether the infant can hear, in terms of perceiving auditory information. Inspite of this major limitation, the growing acceptance of brainstem electrical potentials in clinics and laboratories throughout the world attests to their increasing importance especially in the assessment of hearing sensitivity of infants.


Evaluation of a deaf child.

Evaluation of a deaf child. One should be aware that the measurement of hearing is really an evaluation of a child's behavioural responses in a controlled setting to the presentation of various acoustic stimuli. The determination of hearing loss in some children is an ongoing process and may require more than one test session. Difficult to test children should be evaluated with as many techniques as possible for accurate cross checking of test results. Flexibility, patience and understanding and persistence are the watch words of successful hearing testing with children. For the bulk of patients who require audiological assessment however, the test battery (BOA, IMPEDANCE & BSER) approach seems to be the best available present day approach coupled with long term follow ups and careful management.


Effects of interleukin-1 beta and tumor necrosis factor-alpha on osteoblastic expression of osteocalcin and mineralized extracellular matrix in vitro. Osteoblasts play a pivotal role during the bioresponse of bone to agents that stimulate bone resorption and/or inhibit bone formation including hormones, polypeptide growth factors, and cytokines. We examined the cytokines interleukin-1-beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) for their effects on osteoblastic proliferation and development and expression of alkaline phosphate and the osteoblast-specific protein osteocalcin in a mineralizing environment. Primary rat osteoblast-like cells (ROB) and osteoblastic cell lines derived from rat (ROS 17/2.8) and human (MG-63) osteosarcomas were studied. IL-1 beta and TNF-alpha were chosen because of their critical importance during the host response to local inflammatory stimuli. Qualitatively similar two- to threefold inhibition of osteocalcin synthesis by IL-1 beta and TNF-alpha were observed in all three postconfluent bone-forming model systems. Because of the readily measurable concentrations of osteocalcin produced in our culture protocol, it was not necessary to enhance osteoblastic synthesis of osteocalcin by supplementation with 1,25(OH)2-vitamin D3, a treatment which exerts pleiotropic effects on osteoblasts. Under the constraints of our protocol, where alkaline phosphatase and mineralization were already elevated at the 14-day onset of treatment, neither of these phenotypic properties was sensitive to a three-day cytokine exposure. Differences were noted in proliferation, where only TNF-alpha stimulated DNA synthesis in ROB cells, while both cytokines stimulated MG-63 cells. IL-1 beta and TNF-alpha failed to alter ROS 17/2.8 DNA synthesis except at the highest doses (25 pM IL-1 beta and 1 nM TNF-alpha) where inhibition was observed. These results further support the view that cytokine-mediated osteoblastic regulation can be relatively selective.


Adhesive effect of certain cytokines and other perturbants on human neutrophils. Pretreatment of normal human neutrophils with certain cytokines and other mediators caused some of the cells to become adhesive and stick to the plastic (polypropylene) incubation tubes during pretreatment and during the assay for phagocytosis of C3b.IgG-coated microspheres. Often as much as 40% of the cells were adherent to the tubes after the reaction. This sticking of the neutrophils to the plastic tubes was confirmed by increase in cytometer sipping time and by lactic dehydrogenase assay of the suspended cells and of the cells stuck on the sides of the empty incubation tubes. Only those perturbants that caused an up-regulation of C3b receptors (CR1, CD35) and in most cases caused an enhancement of phagocytosis mediated the adhesiveness of the cells. Unless these stuck cells were detached by vigorous flushing with cold buffer containing EDTA, many of the cells were not admitted into the cytometer for determination of the effect of the perturbants on binding and phagocytic capacity of the neutrophils. This observation could have implications regarding the possibility of subpopulations of neutrophils and differences in function of adherent cells versus cells in suspension. In the cases studied there was no appreciable difference between the total binding and phagocytic capacities of the adherent and suspended cells.


Changes in serum glycoprotein glycosylation during experimental inflammation in mice are general, unrelated to protein type, and opposite changes in man and rat: studies on mouse serum alpha 1-acid glycoprotein, alpha 1-esterase, and alpha 1-protease inhibitor. Using the method of crossed affinity immunoelectrophoresis with concanavalin A in combination with digital image processing, unrelated serum glycoproteins from normal mice and from mice undergoing an experimentally induced inflammation were analyzed for their carbohydrate-derived microheterogeneity profile. This profile changed in a generalized way in mouse serum samples taken at various time intervals after the initial induction of inflammation. The changes are not related to the acute-phase behavior of the protein itself (be it positive, negative, or nonreacting), and they are opposite to the changes in microheterogeneity profiles reported previously for glycoproteins during the acute phase in human and rat sera. These findings are discussed in the context of the biosynthetic control of glycoprotein glycosylation during the acute-phase response.


Loose stools in the early neonatal period. During a two-year period, the various factors associated with loose stools in the early neonatal period were studied among hospital born babies. Low birth weight babies had a lower incidence of non-infective loose stools when compared to neonates with a birth weight of more than 2500 g (p < 0.001). Newborns delivered by Cesarean section (p < 0.001) and those born to women with more than two children (p < 0.02) had a greater frequency of loose stools. Initiation of supplementary feeding and administration of antibiotics were important factors in causing loose stools. Bacterial etiology could be found only in 9.3% of newborns having loose stools. A later onset of loose stools was noted in those, whose stool culture grew bacterial organisms. Only nine newborns with loose stools required antibiotic therapy. Although loose stools were less common among low birth weight babies, they often required treatment with antimicrobials. None of them developed any complications. Since majority of them are non-bacterial and non-infective, great caution must be exercised before administering antibiotics to newborns with loose stools.


Congenital adrenal hyperplasia: experience at Calcutta. Eight patients (7 females, 1 male) with congenital adrenal hyperplasia (CAH), were seen over a 24-month period beginning from March 1988. Seven patients had 21 hydroxylase (21-OH) deficiency while one had 11 beta hydroxylase deficiency. Of the 7 patients with 21-OH deficiency, 3 were of the salt losing (SL-CAH), and 4 were of the non-salt losing (NSL-CAH) type. The patients with NSL-CAH were diagnosed by their elevated 17-hydroxyprogesterone (17-OHP) levels. The 3 cases with SL-CAH were diagnosed on the basis of ambiguous external genitalia, typical electrolyte picture, normal female internal genitalia, sex chromatin and response to steroids. In one patient post-ACTH 17 OHP was alter measured. All 3 patients with SL-CAH were assigned the male sex. Sex reassignment was advised for two children; one accepted the advice and the child is doing well; one family did not accept sex reassignment and the child died. One patient died due to non-availability of fludrocortisone. Six patients are under follow-up. All are doing well except one patient with NSL-CAH who started treatment late. We conclude that a high index of suspicion, early diagnosis and meticulous patient education are the key features of successful management of CAH in India.


Nutritional disorders in adolescent girls. Four hundred and fifty four adolescent girls (11-18 years) were screened for nutritional disorders by anthropometry (weight, height and triceps skinfold measurements), clinical examination and hemoglobin estimation. Of these, 56% belonged to high socio-economic groups (Group A) and the rest (44%) to lower middle class (Group B). A large number of girls from Group B were undernourished (35.5% had weight/height2 less than the fifth percentile of reference standard) stressing the need for nutritional screening, nutrition and health education. Obesity was prevalent in 3.1% of Group A adolescents. Goitre grade I or more was observed in a high proportion of Group B girls, stressing the need for continued consumption of iodized salt in Delhi. Anemia appears to be a major health problem in adolescent girls in both groups (47, 56% in Groups A and B, respectively) underlying the ned for iron supplementation along with health education.


Per rectal diazepam therapy in convulsive disorders. One hundred and twenty children with persistent convulsions (lasting > or = 10 min) were treated with per rectal diazepam (dosage: 0.2 to 0.7 mg/kg/dose). Another group of 100 age matched children with convulsions, along with those who did not respond to rectal therapy were given intravenous diazepam in a dosage of 0.2 to 0.3 mg/kg/dose. Rectal treatment was effective in 80.83% cases while intravenous diazepam was effective in 90% cases which is statistically just significant (p < 0.05). No significant difference was observed in the efficacy of two routes of administration in controlling convulsions of different clinical types and various etiological groups (p < 0.05), except for primary generalized type where intravenous route was more effective than the rectal one (p < 0.05). No significant side-effect was observed with rectal therapy. Among the 23 (19.17%) children in whom rectal therapy failed, 12 (10%) responded to intravenous diazepam while the remaining 11 (9.17%) cases were resistant to both routes of administration.


Kerosene oil poisoning--a childhood menace. This study documents 3-year retrospective analysis of accidental kerosene oil poisoning in 70 children with regard to clinical profile, radiological changes and outcome. About 77% of cases were between 1 and 3 years old. Fifty children (71.4%) developed significant symptoms, with onset soon after to within 10 hours of ingestion. These included breathlessness (55.7%), fever (47.1%), cough (31.4%), restlessness (25.7%) and abdominal distension (15.7%). Chest X-rays were obtained in 65 children. Abnormal radiographs were seen in 45 (69.2%) children with right basal infiltrates being the commonest picture (21.4%). Ingestion of more than one ounce of kerosene oil adversely affected the clinical and radiological profile. Severely malnourished children had extensive radiological changes and poorer clinical outcome. One case developed myocarditis, a complication which has not been reported to the best of our knowledge. Mortality rate was 4.3%. All deaths occurred within 48 hours of admission.


Psychosocial study of leukemic children and their parents. Psychosocial assessment was carried out in 35 children with acute lymphatic leukemic, an equal number with non-leukemic chronic illness and their parents. Psychological dysfunction existed more frequently in parents of leukemic children. Depression as an initial reaction on being conveyed the diagnosis, was seen in 85.8% whilst anger was observed in 42.8%. Majority (89.7%) entertained doubts about whether a correct diagnosis has been established. Understanding of the disease, its possible course and the need for prolonged treatment, was appreciated by about 65.7% of parents. The disease imposed serious social, financial and occupational burdens on the family. Measures used to cope with such stresses included meeting close friends and relatives and finding solace in religious activities. When comparing psychopathology in leukemic children with that in chronically ill non-leukemic counterparts, significant differences were observed in certain specific syndrome scores. Conduct disorder, anxiety, depression and psychotic symptoms were more prevalent in leukemic children. This study emphasizes the necessity of active psychosocial intervention in the total care of childhood leukemia.


Cerebral palsy.

Cerebral palsy. The clinical pattern and etiology of 544 cases of cerebral palsy were studied retrospectively. Of these cases, 354 (65.1%) were males. Four hundred and ninety seven (91.4%) cases were of spastic type. Hypotonic, ataxic and athetoid cerebral palsy were observed in 5.5, 1.5 and 1.3% cases, respectively. There was one case each of tremor and mixed type. In the spastic group, quadriplegia comprised the maximum number of cases (34.9%). Hemiplegia (28.7%) and diplegia (21.9) were also common. Mental retardation was found in 47.2%, while speech impairment was observed in 37% cases. Other handicaps included visual (9%), seizures (8.8%), and auditory handicap (2.9%). The etiological factors were prenatal in 7.7% cases, natal in 43.8% cases and postnatal in 26.1% cases. More than one etiological factor was observed in 14.5% cases, while in 7.9% cases, no apparent cause could be found.


Thyroid dysfunction in multi-transfused iron loaded thalassemia patients. Seventy-two transfusion-dependent iron loaded thalassemia patients were investigated for thyroid dysfunction by estimating circulating thyroid hormones (T4 and T3) and basal thyroid stimulating hormone (TSH). They were also evaluated for their liver function (biochemically) and iron overload by estimating serum ferritin. Thyroid failure (hypothyroidism) was documented in 14 patients (19.4%). In all, 3 groups were seen, i.e. Group 1: Normal T4, T3, TSH (58 patients: 80.6%); Group 2: Compensated hypothyroidism characterized by normal T4, T3 and raised TSH (9 patients: 12.5%); Group 3: Decompensated hypothyroidism characterized by decreased T4 and increased TSH (5 patients: 6.9%). Interestingly, impaired thyroid function could not be correlated with age, amount of blood transfused, liver dysfunction or degree of iron overload. It is postulated that an inter-play between chronic hypoxia, liver dysfunction and iron overload may be responsible for the thyroid damage.


Smoking-related DNA adducts in human gastric cancers. DNA was extracted from the tumour tissue of 26 patients (18 smokers, 8 non-smokers) undergoing surgery for gastric cancer, and analyzed for the presence of DNA adducts by the 32P-post-labelling method. Adducts were detected in all samples tested, and adduct levels ranged from 2 adducts/10(8) nucleotides to 60 adducts/10(8) nucleotides. In male subjects, adduct levels were significantly greater in the DNA of smokers than in that of non-smokers. These results support epidemiological data suggesting that smoking is a risk factor for gastric cancer.


Cell kinetics evaluation of colorectal tumors after in vivo administration of bromodeoxyuridine. Although several biomarkers have been tested, Dukes' (or TNM) stage at diagnosis is still considered the only prognostic factor of clinical relevance in colorectal cancer. Among the various biomarkers, the fraction of cells engaged in DNA synthesis has been extensively investigated as an indicator of tumor aggressiveness. Bromodeoxyuridine (BUdR) is a non-radioactive thymidine analogue which is incorporated into DNA during the S-phase of cycling cells. In order to evaluate the relationships between cell kinetics and morphologic variables, 500 mg of BUdR were given i.v. to 46 patients with colorectal cancer prior to surgery. After operation, a large tumor sample was taken and processed for immunohistochemical detection of BUdR-labeled cells in various regions of the neoplasm and in normal colorectal mucosa. Smaller superficial tumor specimens were also incubated with 3H-thymidine (3H-TdR) for the autoradiographic identification of labeled cells. In the 43 evaluable tumors, the overall BUdR labeling index (BLI, percent of labeled cells) was significantly higher in carcinoma (20.30 +/- 0.86%, SEM) than in normal colonic mucosa (6.51 +/- 0.49%). BLIs in central and peripheral regions of carcinoma were closely correlated (r = 0.48, p = 0.003). In 21 neoplasms a high correlation between overall BUdR and 3H-TdR labeling index in the same tumor was observed (r = 0.57, p = 0.007). No evident association between overall BLI and clinical or morphologic parameters of the tumor was seen, including number of capillaries and ras-p21 protein expression. We conclude that BUdR immunostaining after in vivo administration of BUdR is a simple method for studying cell kinetics in various regions of colorectal cancer. BUdR labeling data are comparable to those obtained with in vitro incorporation of 3H-TdR.


EGF receptor expression in primary laryngeal cancer: correlation with clinico-pathological features and prognostic significance. Epidermal-growth-factor-receptor(EGFR) expression was evaluated in 103 primary laryngeal tumors and in 42 normal laryngeal tissue specimens. Significantly higher EGFR levels were found in cancer specimens than in normal mucosa (p = 0.0053). EGFR expression did not correlate with age, tumor localization, T classification, cervical-lymph-node involvement or type of surgery, whereas it was higher in poorly differentiated tumors (G3) than in well/moderately differentiated (G1-G2) tumors (p < 0.05). Follow-up data were available for 74 patients. When EGFR status and the most important clinico-pathological characteristics were submitted to univariate analysis, tumor localization, type of surgery and EGFR status were found to be significantly correlated with disease-free survival. The 24-month disease-free survival rate was 58% for EGFR+ cancer patients and 82% for EGFR- ones. With multivariate analysis, only EGFR status and tumor localization were identified as significant independent prognostic parameters. Data reported here suggest that high EGFR levels may identify a sub-set of laryngeal-cancer patients with a particularly unfavorable prognosis.


The p53 tumor-suppressor gene and ras oncogene mutations in oral squamous-cell carcinoma. The frequencies of mutations in the p53 tumor-suppressor gene and ras proto-oncogenes were investigated systematically in surgically resected oral squamous-cell carcinomas (SCCs) using single-strand conformation polymorphism (SSCP) and/or dot-blot hybridization analysis of DNA fragments which had been amplified by the polymerase chain reaction (PCR). p53 gene mutations, within the region of exons 5 to 8, were detected in 17 out of 27 (63%) tumor specimens. The role of p53 mutations in cell-line establishment was investigated. p53 gene mutations were detected in 5 out of 6 tissue samples from which cell lines were established and in 4 out of 5 specimens from which cell lines could not be established, suggesting that the presence of p53 gene mutations is not by itself sufficient for cell-line establishment. Tumor samples were also analyzed for point mutational activation of the ras proto-oncogenes. One out of 30 (3%) tumors showed an activating point mutation in codon 12 of H-ras, this being consistent with reports from Europe and USA but not with any from India. Compared to frequencies of the other genetic changes so far reported for oral SCC, the p53 mutations have been observed most often to undergo genetic change. p53 gene mutation is thus intimately involved in the genesis of oral SCC and consequently should be useful as a marker for the diagnosis of this neoplasm.


Prognostic relevance of serum hyaluronan levels in patients with breast cancer. The serum hyaluronan (HA) level of 238 women with breast cancer was measured by means of a specific radiometric assay. The results show no significant increase in serum HA when compared to levels in 120 control sera. A number of prognostic factors were evaluated including stage of disease, lymph-node involvement, tumour size, histology and presence of oestrogen and progesterone receptors in the tumour. No correlation was found with serum HA concentration and we conclude that serum HA level is of no prognostic significance in breast cancer.


Neutralizing activity of human antibodies against the structural protein of human T-cell lymphotropic virus type I. We have identified and mapped the regions responsible for neutralization in the human T-cell leukemia virus type I (HTLV-I) structural proteins by using region-specific human antibodies derived from seropositive blood donors. We have obtained 18 kinds of region-specific antibody (2 in the p19 gag, 10 in the gp46 env and 6 in the gp21 env proteins) from seropositive human plasma by means of an affinity column coupled with the synthetic peptides corresponding to the antigenic regions of the HTLV-I structural proteins. These antibodies were highly specific in ELISA using synthetic peptides as an antigen. Subsequently, we examined the neutralizing activity expressed by the inhibition of virion-induced syncytium formation by region specific antibodies. Twelve of 16 antibodies derived from the env protein were able to inhibit syncytium formation induced by co-cultivation of 8C cells with HTLV-I antigen-positive T cells. The antibodies derived from the p19 gag protein and the seronegative plasma used as the control showed no significant activity. The sequences recognized by the 10 neutralizing antibodies were sites corresponding to amino acids 20 to 49, 89 to 115, 136 to 160, 175 to 199, 213 to 236, 235 to 254, 277 to 292, 332 to 352, 350 to 386, 382 to 403, 426 to 448 and 458 to 488 from the amino terminal of the env protein. These observations suggest that the neutralizing epitopes were widely distributed in the env proteins of HTLV-I.


Establishment of a murine model of malignant mesothelioma. Malignant mesothelioma (MM) is an aggressive tumour of the serosal cavities which is associated with previous asbestos exposure and is generally found to be resistant to conventional forms of therapy. Adequate scientific and clinical assessment of this disease has been severely limited by the relatively low incidence of mesothelioma and the lack of representative cell lines and animal models. The purpose of this study was to develop an asbestos-induced murine model of MM both as an in vivo-passaged malignancy and as in vitro-established cell lines. Such a model system would be invaluable for use in the study of various cellular, molecular and genetic aspects of the disease, and for the pre-clinical evaluation of potential therapeutic agents. BALB/c and CBA mice were injected intraperitoneally with crocidolite asbestos. Seven to 25 months after exposure, 35% of the mice developed mesothelioma (5 BALB/c, 9 CBA), as determined by standard cytological and histological parameters. From these primary tumours, 12 continuously growing cell lines (5 BALB/c, 7 CBA) were established in culture. All have been confirmed as mesothelioma by cytological and ultrastructural (electron microscopy) analyses. These lines have been in culture for 7 to 24 months and have achieved passages above 32 (range 32 to 106). As in the human disease, the murine mesothelioma lines vary in their morphology and growth rates (doubling times ranging from 14 to 30 hr). All cell lines produced tumours when injected into syngeneic mice.


Growth of Hodgkin cell lines in severely combined immunodeficient mice. No animal model exists for the in vivo growth of Hodgkin's-lymphoma-derived cells. Neither unmanipulated Hodgkin's-disease(HD)-derived cell lines nor primary biopsy tissue could be grown in nude mice. Since the severe combined immunodeficient (SCID) mouse has been reported to be a better recipient for transplanted human lymphatic tissue than the nude mouse, we tested whether SCID mice provide suitable conditions for the in vivo growth of HD cell lines. Tumorigenicity of HD cells was tested in untreated and pre-treated SCID mice and in another combined immunodeficient mouse strain, beige/nude/X-linked immunodeficient (BNX) mouse. SCID mice supported in vivo growth of the 6 HD cell lines tested (L428, L540, L591, DEV, HD-LM2, KM-H2). Only one of the 6 lines (DEV) was tumorigenic in BNX mice. No HD cell line proliferated in T-cell-deficient nude mice. Thus, in vivo growth of HD cell lines appeared to be related to the degree of host immunodeficiency. Additional growth supportive treatments such as fibrosarcoma co-transplantation, intraperitoneal mineral oil injection or immunosuppressive pre-treatment (anti-asialo-GMI-antibody injection) permitted growth of 3 additional HD cell lines in BNX mice. The immunophenotype and karyotype of explanted graft cells were identical to the original cell lines. Our experiments describe an effective and reproducible xenograft model for growth of Hodgkin's-disease-derived cell lines. This may be of value for elucidating the growth characteristics of Hodgkin's-lymphoma-derived cells as well as for testing new therapeutic regimens.


Antibody against p58 surface antigen of RA-2 rat rhabdomyosarcoma cells inhibits their metastatic activity. Antibodies generated against the whole membrane preparation isolated from rhabdomyosarcoma RA-2 cells were shown by immunoblotting and immunoaffinity chromatography to recognize 58-kDa polypeptide, p58. The latter was confirmed to be a surface molecule in a test of radioiodination of RA-2 membrane by lodogen. The antibodies added to a suspension of RA-2 cells before their inoculation into rats decreased metastatic activity 50-fold without any noticeable influence on RA-2 proliferation level and viability. The data indicate that masking of p58 surface antigen by antibodies could make RA-2 cells unable to form experimental metastases in lung. We suggest that p58 may participate in the specific recognition by RA-2 of lung endothelial cells.


DOK, a cell line established from human dysplastic oral mucosa, shows a partially transformed non-malignant phenotype. There are many reports of cell lines being established from human oral squamous-cell carcinomas but apparently none of cell lines from dysplastic or "pre-malignant" oral mucosa. We describe here the isolation and characterization of a cell line, DOK (dysplastic oral keratinocyte), from a piece of dorsal tongue showing epithelial dysplasia. The tissue was obtained from a 57-year-old man who was a heavy smoker prior to the appearance of a white patch on his tongue. Eleven years later a squamous-cell carcinoma developed at the site and was excised. Subsequently the remaining dysplasia was removed, and it was from a piece of this that the primary cell cultures which eventually gave rise to DOK were initiated. The DOK line has been single-cell cloned and is apparently immortal. It grows in the absence of 3T3 feeder cells, is anchorage-dependent for growth and is non-tumorigenic in nude mice. The keratin profile of the cells shows a striking similarity to that of the original tongue dysplasia. The karyotype of DOK is aneuploid and complex. By PCR and oligonucleotide hybridization on dot blots, codons 12, 13 and 61 of Ha-ras, Ki-ras and N-ras in DNA extracted from DOK cells were shown to be normal. Immunohistochemistry showed no abnormal, i.e., elevated expression of the onco-suppressor protein p53. Because of its origin and partially transformed phenotype, DOK presents an opportunity to study whether specific carcinogens associated with tobacco and areca nut can cause malignant transformation of oral keratinocytes in vitro.


Protection by muscimol against gastric carcinogenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine in spontaneously hypertensive rats. The effects of prolonged administration of the gamma-aminobutyric acid receptor agonist muscimol on enhanced induction of gastric carcinogenesis by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in spontaneously hypertensive rats (SHR), and on the norepinephrine concentration in the gastric wall and the labeling index of gastric mucosa were investigated. SHR and normotensive Wistar Kyoto (WKY) rats as controls were given a solution of MNNG (25 micrograms/ml) for 25 weeks and then i.p. injections of 0.5 mg/kg body weight of muscimol every other day. In control WKY rats, gastric cancers were found in 1 (7%) of 14 rats examined at week 52. In SHR treated with NaCl solution only, the incidence of gastric cancers was significantly increased to 50% compared with that in control WKY rats. However, treatment of SHR with muscimol significantly increased its incidence to 12% compared with the value in SHR treated with NaCl solution only. The norepinephrine concentration in the gastric wall and the labeling index of the gastric mucosa were significantly greater in SHR than in WKY rats. Prolonged administration of muscimol to SHR significantly reduced the norepinephrine concentration in the antral portion of the gastric wall or the labeling index of the antral epithelial cells. These findings indicate that long-term treatment of SHR with muscimol attenuated the enhancement of gastric carcinogenesis in SHR.


Analysis of factors associated with the tumorigenic potential of 12 tumor clones derived from a single rat colon adenocarcinoma. We analyzed several factors which could influence the immunogenicity of colon tumor cells, using a series of clones derived from a single chemically induced rat adenocarcinoma cell line. These clones display variable tumorigenic potential in syngeneic immunocompetent animals, and it has been established that in this model the tumorigenicity of the cells depends on their ability to escape immune surveillance. The results show an absence of relationship between tumorigenicity and expression of MHC-class-I antigens, cell adhesion to rat fibroblasts or fibroblast extracellular matrix. The secretion of latent and active TGF beta I appeared to be quite variable from one clone to the other, but was unrelated to tumorigenicity. Unexpectedly, some regressive clones produced elevated levels of this cytokine, suggesting that in this model, spontaneous secretion of TGF beta I is not sufficient to impair the immune system of the host. In contrast, the more tumorigenic clones were more resistant than less tumorigenic ones to cytotoxicity mediated by NK or LAK cells. They also showed arrest of cell proliferation after reaching confluence, something not observed in the less tumorigenic clones. Finally, the strongest relationship with tumorigenicity was found for expression of blood-group carbohydrate antigens. Increased expression of blood-group-H antigen and, conversely, decreased expression of beta-galactoside precursors of this antigen correlated with increased tumorigenicity.


Functions of estrogens and anti-estrogens in the rat endometrial adenocarcinoma cell lines RUCA-I and RUCA-II. Inbred rats of the DA/Han and BDII/Han strains have been proposed as suitable model systems for studying hormonal carcinogenesis, because they die mainly from hormone-dependent endometrial adenocarcinoma. Here we characterize the RUCA-I cell line derived from an endometrial adenocarcinoma of an inbred DA/Han rat and the RUCA-II cell line derived from an endometrial adenocarcinoma of an inbred BDII/Han rat. The RUCA-I cell line, if transplanted to the neck of female DA/Han rats, gives rise to endometrial adenocarcinomas at the ectopic site. The morphology of these ectopically grown tumors is predominantly of the moderately differentiated sub-class. In contrast, ectopic tumor growth of the RUCA-II cell line can be observed only if cells are transplanted to athymic nude mice. Biochemically, both cell lines are characterized by the stable expression of estrogen receptors. However, no statistically significant mitotic response of RUCA-I and RUCA-II cells to estradiol was measurable, and no induction of expression of the progesterone receptor by estradiol was detectable, although estradiol transformed the estrogen receptor into its stable DNA-binding state. In contrast, the rate of proliferation of RUCA-I but not of RUCA-II cells was reduced in the presence of 10(-6) M tamoxifen. From these results we conclude that (i) both cell lines, RUCA-I and RUCA-II, represent a new and promising endometrial tumor model; (ii) the mechanism of the hormone-dependent growth regulation of RUCA-I and RUCA-II cells is obviously impaired; (iii) the RUCA-I cell line appears to be a suitable model system for the study of molecular aspects of estrogen- and tamoxifen-dependent gene expression.