• Volume 21, Issue 4

      June 1996,   pages  445-575

    • Suppression of tumourogenicity and overexpression of cell adhesion molecules in AK-5 cells transfected with wild-type p53 gene

      Ashok Khar B V V Pardhasaradhi

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      We have previously shown overexpression of p53 and rearrangement of the p53 gene in AK-5 tumour. In order to study the role of p53 in AK-5 tumourogenicity, we introduced wild-type p53 in AK-5 cells. We have shown suppression of tumourogenicity in AK-5 cells after transfixion with wt p53. In one of the transfected clones 3B4, there was complete loss of tumour forming ability. Clone 3B4 also showed cellular aggregation which correlated well with the higher expression of cell adhesion molecules like fibronectin and hyaluronectin. These observations demonstrate tumour suppression in AK-5 after introduction of wt p53.

    • Identification and partial purification of a human natural killer cell proliferation-inducing factor

      Queen B Saxena Rajiv K Saxena Ronald H Goldfarb Ronald B Herberman Theresa L Whiteside

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      Culture supernatants of Concanavalin A activated human peripheral blood mononuclear cells were found to contain a factor which induced proliferative response in normal peripheral blood mononuclear cells. This proliferation-inducing factor specifically induced and sustained proliferation of purified human NK cells but not of T or B cells. Although interleukin 2 (IL12) also has proliferation-inducing effects on NK cells, the partially purified proliferationinducing factor preparations contained no measurable IL2 contamination. Moreover, neutralizing anti-IL2 antibodies did not block the growth effect of proliferation-inducing factor on purified human NK cells. Other cytokines which were tested, including IL4, IL6, IL7, IL12, TNF and IFN, were all found to be inactive in the proliferation-inducing factor assay. While proliferation-inducing factor by itself had no effect on T-cell proliferation, IL2-induced proliferation of T cells was significantly enhanced in the presence of proliferation-inducing factor, as was IL2-induced NK-cell proliferation. NK cells could be maintained in culture for at least a month in the presence of proliferation-inducing factor alone, but the cells lost their cytolytic activity after 3–4 weeks in culture. Addition of IL2, to NK cells which had been cultured in the presence of proliferation-inducing factor, restored their cytotoxicity. Proliferation-inducing factor activity was partially purified on an anion exchange HPLC column. The molecular weight of proliferation-inducing factor appeared to be about 10 kDa, based on its elution profile on a sizing HPLC column. Our results indicate that proliferation-inducing factor is a novel NK-cell proliferationinducing factor.

    • Qualitative and quantitative detection of aflatoxin B1 in poultry sera by enzyme-linked immunosorbent assay

      C P S Sekhon Jyotika Kapur S S Sodhi S K Jand

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      An indirect competitive inhibition type enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of aflatoxin B1, in poultry sera. Preincubation of aflatoxin B1, samples with the antibody prior to competition yielded better results in terms of higher sensitivity. After competition, amount of antibody bound to solid phase was measured by incubation with anti-rabbit immunoglobulins coupled with horse raddish peroxidase. Intensity of colour decreased as the amount of free aflatoxin B1, increased. Final detection of aflatoxin B1, was made by (i) visual comparison with standard aflatoxin B1 using dot-ELISA (qualitative) and (ii) by plate-ELISA, where optical density was measured at 492 nm (quantitative). Plate-ELISA was more sensitive than dot-ELISA, with sensitivity limits being 100 fg and 1 pg per 10 μl, respectively. However, due to ease and speed of performance, dot-ELISA has greater potential as a test for the diagnosis of mycotoxicosis at the field level.

    • Insights into mechanism of tumour promotion by mezerein

      Nandini Rudra Neeta Singh Malaya Gupta

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      The objective of this study was to gather insights and compare the mode of action of the non phorbol, diterpene mezerein with the phorbol ester, phorbol-12-myristate-13 acetate, in normal and transformed cells. Both phorbol-12-myristate-13 acetate and mezerein are shown to activate the signal transduction pathways involving post translational modification of proteins by poly ADP-ribosylation and by protein kinase C, but to varying extents and showed different time kinetics and cell type differences. Multiple nuclear proteins, especially histones H3d, A24 and HI served as acceptors of poly ADP-ribose in response to PMA in both NIH 3T3 and HDCS cells whereas H1 and H2B were the major acceptors in case of mezerein treatment, similarly in both NIH 3T3 and HDCS cells. The results suggest an epigenetic mechanism (s) in tumour promotion by mezerein.

    • Biodegradation of commercial linear alkyl benzenes byNocardia amarae

      Manju Bhatia H Devendra Singh

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      Laboratory degradation studies of two indigeneously produced linear alkyl benzenes byNocardia amarae MB-11 isolated from soil showed an overall degradation of linear alkyl benzenes isomers to the extent of 57–70%. Degradation of 2-phenyl isomers of linear alkyl benzenes was complete and faster than that of other phenyl position (C3–C7) isomers which were degraded to the extent of 40–72% only. Length of alkyl side chains (C10–C14) had little or no impact on the degradation pattern. Major metabolities detected were 2-, 3-and 4-phenyl butyric acids, phenyl acetic acid and cis, cis-muconic acid. Minor metabolites weretrans-cinnamic acid, 4-phenyl 3-butenoic acid and 3-phenyl pentanoic acid along with two unidentified hydroxy acids. On the basis of the formation pattern of these metabolities, three catabolic pathways of linear alkyl benzenes isomers inNocardia amarae MB-11 were postulated. All the phenyl position (C2–C7) isomers of C10, C12, and C14 linear alkyl benzenes along with 3-phenyl and 5-phenyl isomers of C11 and C13 linear alkyl benzenes were degraded viacis,cis-muconic acid pathway. Other phenyl position isomers of C11 and C13 linear alkyl benzenes with phenyl substitution at even number carbon atoms were principally degraded via phenyl acetic acid pathway whiletrans-cinnamic acid formation provided a minor pathway

    • Assessment of luteal rescue and desensitization of macaque corpus luteum brought about by human chorionic gonadotrophin and deglycosylated human chorionic gonadotrophin treatment

      N Selvaraj R Medhamurthy S G Ramachandra M R Sairam N R Moudgal

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      The objective of the current study was to investigate the mechanism by which the corpus luteum (CL) of the monkey undergoes desensitization to luteinizing hormone following exposure to increasing concentration of human chorionic gonadotrophin (hCG) as it occurs in pregnancy. Female bonnet monkeys were injected (im) increasing doses of hCG or dghCG beginning from day 6 or 12 of the luteal phase for either 10 or 4 or 2 days. The day of oestrogen surge was considered as day ‘0’ of luteal phase. Luteal cells obtained from CL of these animals were incubated with hCG (2 and 200 pg/ml) or dbcAMP (2.5,25 and 100 M) for 3h at 37°C and progesterone secreted was estimated. Corpora lutea of normal cycling monkeys on day 10/16/22 of the luteal phase were used as controls. In addition thein vivo response to CG and deglycosylated hCG (dghCG) was assessed by determining serum steroid profiles following their administration. hCG (from 15–90 IU) but not dghCG (15-90 IU) treatment in vivo significantly (P < 0.05) elevated serum progesterone and oestradiol levels. Serum progesterone, however, could not be maintained at a elevated level by continuous treatment with hCG (from day 6–15), the progesterone level declining beyond day 13 of luteal phase. Administering low doses of hCG (15-90 IU/day) from day 6–9 or high doses (600 IU/day) on days 8 and 9 of the luteal phase resulted in significant increase (about 10-fold over corresponding control P < 0.005) in the ability of luteal cells to synthesize progesterone (incubated controls) in vitro. The luteal cells of the treated animals responded to dbcAMP (P < 0.05) but not to hCC added in vitro. The in vitro response of luteal cells to added hCG was inhibited by 0,50 and 100% if the animals were injected with low (15-90 IU) or medium (100 IU) between day 6–9 of luteal phase and high (600 IU on day 8 and 9 of luteal phase) doses of dghCG respectively; such treatment had no effect on responsivity of the cells to dbcAMP. The luteal cell responsiveness to dbcAMP in vitro was also blocked if hCG was administered for 10 days beginning day 6 of the luteal phase. Though short term hCG treatment during late luteal phase (from days 12—15) had no effect on luteal function, 10 day treatment beginning day 12 of luteal phase resulted in regain ofin vitro responsiveness to both hCG (P < 0.05) and dbcAMP (P < 0.05) suggesting that luteal rescue can occur even at this late stage. In conclusion, desensitization of the CL to hCG appears to be governed by the dose/period for which it is exposed to hCG/dghCG. That desensitization is due to receptor occupancy is brought out by the fact that (i) this can be achieved by giving a larger dose of hCG over a 2 day period instead of a lower dose of the hormone for a longer (4 to 10 days) period and (ii) the effect can largely be reproduced by using dghCG instead of hCG to block the receptor sites. It appears that to achieve desensitization to dbcAMP also it is necessary to expose the luteal cell to relatively high dose of hCG for more than 4 days

    • Prolactin augments luteinizing hormone binding to rat Leydig cells in serum-free culture

      Minoru Takase Kazuyoshi Tsutsui Seiichiro Kawashima

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      Effect of prolactin on the testicular luteinizing hormone binding was studied in a serum-free culture system. By the collagenase digestion of decapsulated testes taken out from 25-day-old rats, Leydig cells were isolated and cultured for 7 days in DME/F12 (1:1) medium supplemented with insulin, transferrin, epidermal growth factor, and gentamicin. The cultured cells exhibited the 3β-hydroxysteroid dehydrogenase activity. Hill plots constructed from the data of competition experiment showed that the dissociation constant (Kd) was 0.33 × 10–10M. The Kd value was approximately the same as the known value for the rat testicular homogenates. When the Leydig cells were cultured with ovine prolactin for the last 3 days of 7-day culture period, the binding of luteinizing hormone increased to 1.7-fold ofthat in the control group. From these results it is concluded that prolactin acts to up-regulate the binding of luteinizing hormone to rat testicular Leydig cells in serum-free culture

    • Ammonium sulphate induced stress related alterations in the respiratory epithelium of the airbreathing organ of the catfishHeteropneustes fossilis (Bloch)

      V I Paul T K Banerjee

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      In this paper, histopathological changes in the inner lining of the accessory respiratory organ ofHeteropneustes fossilis following exposure to sublethal concentration (0.2 g I–1) of ammonium sulphate (3 mg I–1 total ammonia-N) has been described. The goblet cells show periodic increased followed by decreased secretory activities. Necrosis and shedding of the epithelial cells over the secondary lamellae cause periodic haemorrhages which lead to degeneration and decreased number of secondary lamellae. Subsequently regeneration takes place each time as evidenced by the appearance of inflammatory tissue. Fusion of more than one secondary lamellae is also common. Regeneration also leads to uncontrolled hyperplasia of haphazardly arranged epithelial cells. This hyperplasia causes increased distance of respiratory blood-air barrier in the secondary lamellae, leading to impaired normal aerial respiration

    • Alteration in the acceptor side of photosystem II of chloroplast by high light

      U Dwivedi R Bhardwaj M Sharma

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      The effect of high light on the acceptor side of photosystem II of chloroplasts and core particles of spinach was studied. BothVmax and apparentKm for DCIP were altered in photoinhibited photosystem II core particles. The double reciprocal plot analysis as a function of actinic light showed increased slope in chloroplasts photoinhibited in the presence of DCMU. Exposure of chloroplasts to high light in the presence of DCMU did not protect the chloroplast against high light induced decrease in Fm, level. Further the high light stress induced decrease inFm level was not restored by the addition of DCMU. These results suggest that the high light stress induced damage to chloroplast involves alteration in the binding site forQB on the DI protein on the acceptor side of photosystem II

    • Biomass estimation using satellite remote sensing data—An investigation on possible approaches for natural forest

      P S Roy Shirish A Ravan

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      Vegetation type and its biomass are considered important components affecting biosphere-atmosphere interactions. The measurements of biomass per unit area and productivity have been set as one of the goals for International Geosphere-Biosphere Programme (IGBP). Ground assessment of biomass, however, has been found insufficient to present spatial extent of the biomass. The present study suggests approaches for using satellite remote sensing data for regional biomass mapping in Madhav National Park (MP). The stratified random sampling in the homogeneous vegetation strata mapped using satellite remote sensing has been effectively utilized to extrapolate the sample point biomass observations in the first approach.

      In the second approach attempt has been to develop empirical models with satellite measured spectral response and biomass. The results indicate that there is significant relationships with spectral responses. These relationships have seasonal dependency in varying phonological conditions. The relationships are strongest in visible bands and middle infrared bands. However, spectral biomass models developed using middle infrared bands would be more reliable as compared to the visible bands as the later spectral regions are less sensitive to atmospheric changes

      It was observed that brightness and wetness parameters show very strong relationship with the biomass values. Multiple regression equations using brightness and wetness isolates have been used to predict biomass values. The model used has correlation coefficient of 0.77. Per cent error between observed and predicted biomass was 10.5%. The biomass estimated for the entire national park using stratified and spectral response modelling approaches were compared and showed similarity with the difference of only 4.69%. The results indicate that satellite remote sensing data provide capability of biomass estimation

    • Evaluating second year cropping on jhum fallows in Mizoram, north-eastern India—Phytomass dynamics and primary productivity

      Tawnenga Uma Shankar R S Tripathi

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      Cropping on jhum fallows in north-eartern India is predominantly done for one year in a jhum cycle. If second year cropping is done, expanse of the forest land required for slashing and burning could be reduced significantly. We tested this hypothesis in a young (6 yr) and an old (20 yr) jhum fallow. We also evaluated if the productivity during second year cropping could be alleviated by auxiliary measures such as tilling the soil or application of fertilizers (chemical or farm-yard manure or both in combination). The results demonstrate that the ecosystem productivity (total dry matter production) and economic yield (rice grain production) decline with shortening of jhum cycle. Second year cropping causes a further decline in ecosystem productivity in old jhum field, but not in young jhum field. Economic yield from second year cropping in its traditional form (without any fertilizer treatment) is not much lower than that in the first year, and can be improved further by manuring the soil. Tilling of soil improves neither ecosystem productivity nor economic yield. Different fertilization treatments respond differently; while inorganic manuring enhances ecosystem productivity, a combination of inorganic and organic manuring improves economic yield

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