Comparative efficacy of two standard methods for determination of iron and zinc in fruits, pulses and cereals.

Micronutrients are essential elements needed in small amounts for adequate human nutrition and include the elements iron and zinc. Both of these minerals are essential to human well-being and an adequate supply of iron and zinc help to prevent iron deficiency anaemia and zinc deficiency, two prevalent health concerns of the developing world. The levels of zinc and, iron were measured in the Banana, Papaya, Rice, Finger millet, Soybean and Urdbean. Standard Atomic absorption spectroscopy (AAS) method was also applied to all the samples for zinc and iron analysis and compared with inductively coupled plasma mass spectroscopy (ICP-MS). It was observed that there was no matrix interference affecting the determination of both elements interested in all the samples analyzed. Average concentration relative standard deviation and standard deviation were used for the statistical evaluation of the results for both elements. Correlation coefficient was used as statistical model to compare both the techniques.

Identification of potential accessions of Asparagus racemosus for root yield and shatavarin IV content.

Successful restoration of over exploited species (Asparagus racemosus) depends upon variability, conservation and cultivation. Twelve elite accessions were characterized for fifteen quantitative and qualitative traits for sustainable cultivation and industrial uses. The evaluated accessions varied in morphology, herbage, root yield and shatavarin IV content. The accession DAR-7 was showing maximum herbage yield (1860 and 1850 g plant-1), fresh root weight (36.33 and 37.33 g plant-1), root girth (18.25 and 18.45 cm) and root yield (14.26 and 12.79 kg plant-1) in both the harvesting years. Shatavarin IV content in roots was maximum in DAR-14 (152.06 and 151.72 µg g-1), followed by DAR-28 (81.16 and 83.16 µg g-1). For economic yield accessions DAR-7, DAR-19, DAR-14, DAR-28 were found superior therefore, they may be further used in crop improvement program as valuable selection. In the cropping system they may act as a viable replacement of traditional crops viz., cumin, gram, cotton and groundnut. Asparagus cultivated under high density plantation ensured high economic return (Rs. 4.87 l ha-1year-1) with 3.66 B: C ratio, therefore, it could be considered a high returns substitute for traditional crops.

Mining of Indian wheat germplasm collection for adult plant resistance to leaf rust.

Leaf rust (Puccinia triticina Eriks.) is a fungal disease of wheat (Triticum spp.), which causes considerable yield loss. Adult plant resistance (APR) is one of the most sustainable approaches to control leaf rust. In this study, field-testing was carried out across ten different locations, followed by molecular screening, to detect the presence of APR genes, Lr34+, Lr46+, Lr67+ and Lr68 in Indian wheat germplasm. In field screening, 190 wheat accessions were selected from 6,319 accessions based on leaf tip necrosis (LTN), disease severity and the average coefficient of infection. Molecular screening revealed that 73% of the accessions possessed known APR genes either as single or as a combination of two or three genes. The occurrence of increased LTN intensity, decreased leaf rust severity and greater expression of APR genes were more in relatively cooler locations. In 52 lines, although the presence of the APR genes was not detected, it still displayed high levels of resistance. Furthermore, 49 accessions possessing either two or three APR genes were evaluated for stability across locations for grain yield. It emerged that eight accessions had wider adaptability. Resistance based on APR genes, in the background of high yielding cultivars, is expected to provide a high level of race non-specific resistance, which is durable.

The beta-adrenergic receptor of newborn mouse skin.

Binding of the potent beta-adrenergic antagonist [125I]iodohydroxybenzylpindolol ([125I]IHYP) to particulate preparations from newborn mouse skin was characterized. A number of criteria were used to establish that binding occurred to specific, high affinity beta-adrenergic receptors in the skin preparations. Thus specific binding (that displaced by 10 micrometer concentrations of the beta-adrenergic antagonist (-)propranolol) reached equilibrium in 15--20 min, was saturable (ligand concentration for half-maximal saturation, 0.25 nM) and freely reversible. Stereoselectivity of binding was demonstrated by the observation that displacement of [125I]IHYP by (-)propranolol occurred at concentrations at least 100 times lower than with (+)isoproterenol. Displacement was also observed with the beta-adrenergic agonists (-)isoproterenol, (-)epinephrine and (-)norepinephrine, but not with the alpha-adrenergic antagonist phentolamine.

Decreased accumulation of cyclic adenosine 3',5'-monophosphate in "ischemic" skin after 12-0-tetradecanoyl-phorbol-13-acetate treatment.

The effect of 12-0-tetradecanoyl-phorbol-13-acetate (TPA) on cyclic adenosine 3',5'-monophosphate (cyclic AMP) level in adult mouse skin in response to ischemia was examined. The incubation of skin pieces in a buffered salts medium at 37 degrees C resulted in a rapid accumulation of cyclic AMP. In mouse skin pieces maximum accumulation (about 6 times the basal level) occurred after 2 min incubation and was followed by a rapid decline in the cyclic AMP level. This "ischemic" rise in epidermal cyclic AMP was greatly reduced if skin was used 16 hr after a single application of 17 nmoles of TPA. The effect of TPA on cyclic AMP accumulation in response to ischemia was first observed at 1 hr after TPA treatment and was maximal at 4 hr. The lack of "ischemic" response in TPA-treated skin was not related to an increase in the activity of cyclic AMP phosphodiesterase after TPA application. In addition, the accumulation of cyclic AMP in skin in response to both ischemia and exposure to isoproterenol, adenosine, histamine, or prostaglandin E2 (PGE2) was not observed in skin treated with the tumor promoter TPA.

The effect of ischemia on cyclic adenosine 3',5'monophosphate accumulation in mouse skin.

The incubation of adult mouse skin pieces in a buffered salts medium at 37 degrees C led to a rapid accumulation of cyclic adenosine 3'5-monophosphate (cyclic AMP) in the tissue. In mouse skin maximum accumulation occurred after 2 min incubation; levels reverted to near control levels after a further 7 min incubation. the increase in cyclic AMP contents of the skin pieces was probably not due to the release of materials which activate adenylate cyclase after binding to cellular receptors. Thus, cyclic AMP accumulation was unaffected by the inclusion of alpha- or beta-adrenergic antagonists, or by the pretreatment of adult mouse skin with indomethacin (an inhibitor of prostaglandin synthetase). Furthermore, adenosine, a known activator of epidermal adenylate cyclase, could not be detected in the incubation medium. The functional integrity of epidermal adenylate cyclase was maintained during the cyclic AMP accumulation in response to ischemia. Thus, adenosine, histamine, isoproterenol and prostaglandin E2 (PGE2) augmented the cyclic AMP response. Cyclic AMP accumulation at 37 degrees C was not observed in newborn mouse skin; this lack of cyclic AMP accumulation was probably not due to increased activity of low affinity cyclic AMP phosphodiesterase in newborn mouse skin.

Accumulation of cyclic adenosine 3',5'-monophosphate in adult and newborn mouse skin: responses to ischemia and isoproterenol.

Levels of cyclic adenosine 3',5'-monophosphate in adult mouse skin pieces were rapidly increased on incubation at 37 degrees C, or on exposure to isoproterenol. Accumulation of the cyclic nucleotide under both conditions was greatly decreased in newborn mouse skin, or in adult skin treated with the tumor promotor 12-0-tetradecanoyl-phorbol-13-acetate.

Effects of cholera toxin on ornithine decarboxylase activity in mouse skin.

The subcutaneous injection of cholera toxin into adult mice resulted in a sustained increase in cyclic AMP levels in mouse epidermis after a lag period of about 2 hr. An increase in ornithine decarboxylase activity occurred between 7 and 10 hr, which was maintained for at least 10 hr. The increase in decarboxylase activity was localized to the area of epidermis visually affected by cholera toxin and was unaffected by hypophysectomy, suggesting a direct effect of the toxin on the epidermal cells. The subcutaneous injection of cholera toxin also led to an increase in cyclic AMP levels in newborn mouse skin. In contrast to adult mice, newborn mouse skin contained high basal activities of ornithine decarboxylase in both the epidermal and dermal fractions. The activity in both fractions was markedly decreased following cholera toxin injection. The ability of cholera toxin to induce both epidermal and dermal ornithine decarboxylase activity developed between 10 and 21 days after birth.

Effect of tunicamycin on receptors for tumor promoters.

A progressive decline in the specific binding of [20-3H]phorbol 12,13-dibutyrate ([3H]PDBu) and glycoprotein synthesis was observed following treatment of primary mouse epidermal cells with tunicamycin, a specific inhibitor of dolichol-mediated glycosylation. Following 18 h of treatment, the specific binding of [3H]PDBu was reduced to 33-56% of the control value. The total protein synthesis determined by leucine incorporation into acid-insoluble material was not altered by this antibiotic drug. These results suggest that the receptor for phorbol diesters is, or is functionally linked to, a glycoprotein on the cell surface.

Oocyte retrieval and histological studies of follicular population in buffalo ovaries.

Ovaries (N = 250) from slaughtered buffaloes were collected to study follicular population and compare methods of oocyte retrieval. The number and size of surface follicles were recorded and grouped into different categories. Different sized follicles in relation to oocyte diameter were studied histologically. Yield of oocytes per ovary were less (P < 0.05) from ovaries bearing a corpus luteum (CL). Techniques used for oocyte recovery included slicing, follicle puncture and aspiration. The oocyte recovery rate was greatest (P < 0.05) using slicing. The average number of visible surface follicles was 5.20 +/- 0.97 with mean numbers of 2.5, 1.2, 0.82 and 0.62 per ovary for follicles sized 4, 8, 12 and 12mm respectively. Histological studies revealed large numbers of primordial follicles in prepubertal and atretic follicles in senile buffaloes. They also established a biphasic relationship of growth between oocyte diameter and follicular size.

Clinical profile of pulmonary aspergilloma complicating residual tubercular cavitations in Northen Indian patients.

BACKGROUND: Little is known regarding the clinical profile of Aspergilloma in Indian patients. Such a study was undertaken at Hospital for Chest and TB, Jaipur. MATERIALS AND METHODS: Old, treated patients of pulmonary tuberculosis showing ball like lesion/s inside cavity/ies or a recent thickening of cavity wall were enrolled. Morning sputa samples were collected in the patients who were able to raise sputum and were examined by KOH mount and fungal culture. Serum anti-aspergillus antibodies were estimated in all the patients. Twenty normal healthy subjects were included to serve as control. All patients showing a positive or borderline positive serology were diagnosed as pulmonary aspergilloma (PA group). The remaining patients formed the non-aspergilloma group (Non PA group). RESULTS: A total of 98 study patients could be classified as PA group (54 patients by serology alone, 44 patients by serology as well as sputum culture). The remaining 152 patients were classified as non PA group. Hemoptysis alone or along with other chest symptoms was significantly more common in PA group as compared to non PA group patients (P<0.001), more so in those with ball like lesions. But chest symptoms other than hemoptysis were more common in non PA group. Within the PA group, 21 (13 with ball like lesions and 8 with thickening of cavity wall) had clinical symptoms suggestive of CNPA and two patients (one each with ball like lesions and thickening of cavity wall) had clinical symptoms suggestive of ABPA. CONCLUSION: The clinical profile of pulmonary Aspergilloma in Indian patients is very protean ranging from saprophytic disease to CNPA and less commonly to ABPA.

Elevated circulating levels of matrix metalloproteinase-9 and -2 in patients with symptomatic coronary artery disease.

BACKGROUND: Matrix metalloproteinases (MMP-9 and MMP-2) have been implicated in development of atherosclerosis and plaque rupture in acute coronary syndromes (ACS). AIM: To determine the relationship between circulating MMPs and symptomatic coronary artery disease. METHODS: Plasma levels of MMP-9 and MMP-2 were measured in patients with ACS, stable angina (SA) and in controls, using a quantitative gelatin zymography. These measurements were correlated with markers of systemic inflammation (hs-CRP) in all subjects and myocardial injury (troponin T) in patients with ACS. RESULTS: Plasma MMP-9 in ACS was greater than in SA, and was greater in SA than in controls (P < 0.01 ACS vs SA and controls, P < 0.01 SA vs control). Plasma MMP-2 was significantly greater in ACS than SA or controls (P < 0.01 vs SA and controls). There was strong overall relationship between hs-CRP and MMP-9 (r = 0.65, P < 0.0001) driven by a significant relationship in ACS patients (r = 0.58, P = 0.02), as there was no significant association in SA or controls. A weaker overall correlation was found between hs-CRP and MMP-2 (r = 0.39, P = 0.02), but no significant relationship was present for either of the two patient subgroups or controls. There was no correlation between levels of troponin T and MMP-9, MMP-2 or hs-CRP in ACS patients. CONCLUSION: Quantitative gelatin zymography identifies increased circulating levels of MMP-9 and MMP-2 in patients with symptomatic coronary disease. MMP-9 and MMP-2 are higher in ACS than SA patients and might have use as markers of plaque rupture or instability. The strong relationship between MMP-9 and hs-CRP in ACS patients suggests MMP-9 might be an additional marker and/or consequence of the inflammatory component in ACS.

Specific binding of phorbol ester tumor promoters to intact primary epidermal cells from Sencar mice.

The binding of [20-3H]phorbol 12,13-dibutyrate ([3H]PDB) to intact living epidermal cells in monolayer culture was characterized. At 37 degrees C, the maximum specific [3H]PDB binding (binding displaceable by 30 microM unlabeled PDB) was attained in 15--20 min and was followed by a rapid decrease (down regulation) of radioactivity bound to the cells. The activity lost by the cells during this decrease was found in the incubation medium. Prior exposure of cells to phorbol 12-myristate 13-acetate (PMA; 12-O-tetradecanoylphorbol 13-acetate) but not to phorbol for 2 hr at 37 degrees C caused approximately 55% reduction in the number of measurable binding sites for [3H]PDB. The down regulation was temperature sensitive; there was no loss of radioactivity after 1 hr at 4 degrees C. The specific binding of [3H]PDB at 4 degrees C reached equilibrium in 15--20 min and was saturable and freely reversible. At equilibrium, epidermal cells contained 1.2 x 10(5) binding sites per cell, and binding sites had a KD of 10 nM. Specificity of binding was shown by the observation that the biologically active phorbol esters PMA and 12-deoxyphorbol 13-decanoate inhibited the binding, whereas the inactive parent compound phorbol and the nonphorbol tumor promoter anthralin did not have any effect. The abilities of these compounds to inhibit [3H]PDB binding directly correlates with their tumor promoting activities. Epidermal cells exposed to retinoic acid or fluocinolone acetonide for 24 hr had similar [3H]PDB binding characteristics as untreated cells suggesting that inhibition of tumor promotion induced by these compounds is not mediated through alterations in the phorbol ester binding sites.

Down regulation of specific binding of [20-3H]phorbol 12,13-dibutyrate and phorbol ester-induced differentiation of human promyelocytic leukemia cells.

Binding of [20-3H]phorbol 12,13-dibutyrate ([3H]PDB) to intact human promyelocytic leukemia cells susceptible (HL-60) or resistant (R-35) to phorbol ester-induced differentiation was characterized. Specific binding of [3H]PDB to both HL-60 and R-35 cells at 37 degrees C reached a maximum within 15-20 min. Maximal specific [3H]PDB binding to HL-60 cells was followed by a decline (down regulation) of radioactivity. This down regulation was temperature dependent, because no loss of radiolabel occurred by 1 hr at 4 degrees C. The down regulation of bound [3H]PDB seen in HL-60 cells at 37 degrees C was not observed with R-35 cells. Prior exposure of the HL-60 cells but not of R-35 cells to 1 microM phorbol 12-myristate 13-acetate for 90 min at 37 degrees C caused a marked reduction in the specific binding of [3H]PDB. When [3H]PDB binding was carried out at 4 degrees C, [3H]PDB bound to both cell types in a rapid, specific, and reversible manner. At equilibrium, HL-60 and R-35 cells were found to contain almost the same number of binding sites, which had dissociation constants of about 50 nM, indicating that the failure of R-35 cells to undergo PDB-induced differentiation was not associated with any change in the affinity or in the number of [3H]PDB binding sites. These results indicate that the down regulation of specific [3H]PDB binding may be a crucial early event in the control of phorbol ester-induced terminal differentiation in HL-60 cells. Furthermore, we suggest that such down regulation may be involved in other cellular and biochemical effects of phorbol diester tumor promoters.

Effect of tumor promoters on lipid peroxidation in mouse skin.

Products of lipid peroxidation were measured in mouse epidermis. These were shown to increase with advancing age. Conversely, a decline in these products was observed on treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA), a potent tumor promoting agent. A decline in lipid peroxidation occurred within 4 h after application of 2 micrograms TPA and the maximum effect was seen at 22-24 h. A lesser active tumor promoter, phorbol dibenzoate; and ethyl phenylpropiolate, a purely hyperplastic agent, also lowered lipid peroxidation; while phorbol, a non-promoter, did not show any significant effect. Mezerein, a resiniferonal derivative with weak promoting activity but a potent stage II promoter, appeared to be more potent than TPA in lowering the basal levels of peroxidation. The TPA-induced decrease in lipid peroxidation could be prevented by fluocinolone acetonide, a potent antipromoting and antimitotic agent, but not by retinoic acid and tosylamino-2-phenylethylchlorimethyl-ketone which are relatively potent antipromoting agents but lack antimitotic activity, suggesting that the decrease of lipid peroxidation by tumor promoting agents is possibly related to their mitotic activity. Furthermore, skin papilloma and carcinoma contain lower levels of lipid peroxidation compared to epidermis from the same mice.

Diminution of mouse epidermal superoxide dismutase and catalase activities by tumor promoters.

The effects of phorbol ester tumor promoters and related compounds on superoxide dismutase (SOD) and catalase were examined. The treatment of adult mouse skin with 2 micrograms 12-O-tetradecanoylphorbol-13-acetate (TPA) resulted in a sustained decrease in the basal levels of both SOD and catalase activities in the epidermis. A decline in SOD activity occurred within 3 h after application and the maximum effect was seen at 16--17 h. The decrease in SOD activity was always accompanied by a similar decline in the epidermal catalase activity. The alterations in both enzymes occurred against a high background of enhanced protein synthesis which indicates that the effect of TPA is selective for SOD and catalase. Other tumor promoters such as phorbol 12,13-dibutyrate and the non-phorbol tumor promoter anthraline also lowered the activities of both the enzymes. Mezerein, a resiniferonol derivative with weak promoting activity but a potent stage-II promoter, appeared to be more potent than TPA in lowering the basal levels. These results indicate that damage which favors neoplastic progression could occur in TPA-treated mouse skin due to the accumulation of free radicals resulting from low levels of SOD and catalase activity. In addition, the TPA-caused decrease in the levels of SOD and catalase was not prevented by either retinoic acid, fluocinolone acetonide, tosyl amino-2-phenylethyl chloromethyl ketone, or butylated hydroxytoluene, suggesting that inhibition of tumor promotion by these agents is not mediated through alterations in the levels of enzymatic activities which decrease free radical concentrations.

The reduction of tumor initiating activity and cell mediated mutagenicity of dimethylbenz[a]anthracene by a copper coordination compound.

Cu(II) (3,5-diisopropyl salicylate)2 (CuDIPS) which is an anti-inflammatory copper coordination compound (mol. wt. 503) possessing superoxide dismutase (SOD) activity was tested to determine its effect on 7,12-dimethylbenz[a]anthracene (DMBA)-induced initiation of tumors in mouse skin and on mutagenicity to 6-thioguanine resistance in a mouse keratinocyte mediated Chinese hamster V-79 cell system. A single application of CuDIPS (0.4 mg/mouse) administered at a short interval before DMBA application when followed by 20 weeks of promotion by TPA reduced the mouse skin tumor yield by 55%. When DMBA-induced cell-mediated mutagenesis was tested in the presence of CuDIPS a significant reduction in the number of V-79 6-thioguanine resistant mutants was observed.