Purification of antibodies for the cytokinin N6-(delta 2-isopentenyl) adenosine by affinity chromatography.

Isopentenyl adenosine antibodies useful in the investigations of the "cytokinin" functions of isopentenyl adenosine were purified by affinity chromatography. Using different affinity columns, the antibodies were purified to near complete purity. Analyses of the purified proteins revealed the presence of isopentenyl adenosine binding proteins in normal rabbit serum, which presence supports a suggested role for isopentenyl adenosine and its related compounds in animal cell division in vivo.

Effect of antiserum to luteinizing hormone (LHAS) on the physiology of the epididymis and accessory glands in the albino rat

Rabbit antiserum specific to ovine luteinizing hormone free of contaminating antibodies to nonspecific proteins and FSH was administered to adult, intact rats at a dose of 0.1 and 0.2 ml/day for five days. LHAS had no effect on the weights of the epididymis but decreased their secretory activity to castrate level. Administration of 0.2 ml of LHAS or castration resulted in a marked and comparable reduction in the weights and secretory activity of the accessory glands. LHAS, even at a lower dose (0.1 ml/day), caused a significant reduction in the content of sialic acid in the vas deferons and Cowper's glands. These results are discussed in relation to the factors that regulate the functions of the epididymis and accessory glands.

Toxicity in Cuscuta reflexa Sucrose Content Decreases In Shoot Tips Upon Trehalose Feeding Trehalose

Trehalose, an {alpha},{alpha}-diglucoside, induced a rapid blackening and death of shoot tips of Cuscuta reflexa (dodder) cultured in vitro. The onset of toxic symptom was delayed if any of the several sugars which support the in vitro growth of Cuscuta was supplied with trehalose. The rate of trehalose uptake or its accumulation in the tissue was not affected by sugar cofeeding. The levels of total and reducing sugars declined appreciably in the trehalose-fed shoot tip explants compared to control tissue cultured in absence of a carbon source. This was not due to an increased rate of respiration of the trehalose-treated tissue. In shoot tips cultured in presence of both trehalose and sucrose, the decline in total and reducing sugars was curtailed. There was a marked fall in the level of sucrose; and invertase activity was higher in trehalose-fed shoot tips. The incorporation of label from [14C]glucose into sucrose in the shoot tip explant was reduced as early as 12 h of trehalose feeding. The results suggest that increased utilization of sucrose as well as an inhibition of its synthesis contribute to the drastic fall in the sucrose content upon trehalose feeding

Role of glutamine synthetase in citric acid fermentation byAspergillus niger

The activity of glutamine synthetase fromAspergillus niger was significantly lowered under conditions of citric acid fermentation. The intracellular pH of the organism as determined by bromophenol blue dye distribution and fluorescein diacetate uptake methods was relatively constant between 6•0–6•5, when the pH of the external medium was varied between 2•3–7•0.Aspergillus niger glutamine synthetase was rapidly inactivated under acidic pH conditions and Mn2+ ions partially protected the enzyme against this inactivation. Mn2+-dependent glutamine synthetase activity was higher at acidic pH (6•0) compared to Mg2+-supported activity. While the concentration of Mg2+ required to optimally activate glutamine synthetase at pH 6•0 was very high (≥ 50 mM), Mn2+ was effective at 4 mM. Higher concentrations of Mn2+ were inhibitory. The inhibition of both Mn2+ and Mg2+-dependent reactions by citrate, 2-oxoglutarate and ATP were probably due to their ability to chelate divalent ions rather than as regulatory molecules. This suggestion was supported by the observation that a metal ion chelator, EDTA also produced similar effects. Of the end-products of the pathway, only histidine, carbamyl phosphate, AMP and ADP inhibitedAspergillus niger glutamine synthetase. The inhibitions were more pronounced when Mn2+ was the metal ion activator and greater inhibition was observed at lower pH values. These results permit us to postulate that glutamine synthesis may be markedly inhibited when the fungus is grown under conditions suitable for citric acid production and this block may result in delinking carbon and nitrogen metabolism leading to acidogenesis.

Fluorescent probe and NMR studies of the aggregation of bile salts in aqueous solution

The binding of the fluorescent probes 1-anilino-8-naphthalene sulfonate and dansyl cadaverine to the sodium salts of cholic, deoxycholic and dehydrocholic acids has been investigated. Enhanced probe solubilisation accompanies aggregation. Monitoring of fluorescence intensities as a function of bile salt concentration permits the detection of primary micelle formation, as well as secondary association. The transition concentrations obtained by fluorescence are in good agreement with values determined for the critical micelle concentrations, by other methods. Differences in the behaviour of cholate and deoxycholate have been noted. Fluorescence polarisation studies of 1,6-diphenyl-1,3,5-hexatriene solubilised in bile salt micelles suggest a higher microviscosity for the interior of the deoxycholate micelle as compared to cholate. 1H NMR studies of deoxycholate over the range 1–100 mg/ml suggest that micelle formation leads to a greater immobilisation of the C18 and C19 methyl groups as compared to the C21 methyl group. Well resolved 13C resonances are observed for all three steroids even at high concentration. Both fluorescence and NMR studies confirm that dehydrocholate does not aggregate.

Homoagmatine from Lathyrus sativus seedlings

A new guanidino amine has been isolated from Lathyrus sativus seedlings and chararterized as homoagmatine on the basis of various physico-chemical criteria including IR spectrum and comparison with that chemically synthesized. Homoagmatine is accumulated in the embryos axis while its precursor, homoarginine, is lost from the cotyledons. However, there was a progressive increase in homoarginine content of the embryo axis during development. Since the amine content of the whole seedlings corresponded to nearly 20–25 % of net decrease in homoarginine levels, it is concluded that the catabolism of homoarginine through homoagmatine represents a major pathway of metabolism of the arnino acid.

Increasing anthocyanin content in Queen Garnet plum and correlations with in-field measures

While plums are traditionally bred for fresh fruit traits such as size, sweetness, yield and disease resistance the Queensland Government breeding program for Japanese plum ( Prunus salicina Lindl.) also selected for anthocyanin content to develop a new plum selection named 'Queen Garnet'. When ripe or overripe, it has a near black skin and deep red flesh colour, which when combined, result in exceptionally high anthocyanin content, reaching up to 277 mg/100 g fruit. The skin fraction contributes 36-66% of the total anthocyanin content of fruit. The plum is now being commercially grown to be processed into a range of functional products from food colourants to premium health products. These are sold on the basis of anthocyanin and antioxidant content. Protocols for increasing anthocyanin content have therefore been researched to maximise the total anthocyanin yield rather than fresh fruit weight and taste. The principal approach is through selective harvest of overripe plums high in colour, although post-harvest storage at 21°C results in further anthocyanin synthesis. Modified processing is also required to ensure recovery of anthocyanins from the skin fraction. The plum products have entered testing for assessing health properties beginning with an initial proof of in vivo bioavailability of the anthocyanins.

Increase in β-Ionone, a Carotenoid-Derived Volatile in Zeaxanthin-Biofortified Sweet Corn

Carotenoids are responsible for the yellow color of sweet corn (Zea mays var. saccharata), but are also potentially the source of flavor compounds from the cleavage of carotenoid molecules. The carotenoid-derived volatile, -ionone, was identified in both standard yellow sweet corn (Hybrix5) and a zeaxanthin-enhanced experimental variety (HZ) designed for sufferers of macular degeneration. As -ionone is highly perceivable at extremely low concentration by humans, it was important to confirm if alterations in carotenoid profile may also affect flavor volatiles. The concentration of -ionone was most strongly correlated (R2 > 0.94) with the -arm carotenoids, -carotene, -cryptoxanthin, and zeaxanthin, and to a lesser degree (R2 = 0.90) with the α-arm carotenoid, zeinoxanthin. No correlation existed with either lutein (R2 = 0.06) or antheraxanthin (R2 = 0.10). Delaying harvest of cobs resulted in a significant increase of both carotenoid and -ionone concentrations, producing a 6-fold increase of ?-ionone in HZ and a 2-fold increase in Hybrix5, reaching a maximum of 62g/kg FW and 24g/kg FW, respectively.

Investigation of the microbial metabolism of carbon dioxide and hydrogen in the kangaroo foregut by stable isotope probing

Kangaroos ferment forage material in an enlarged forestomach analogous to the rumen, but in contrast to ruminants, they produce little or no methane. The objective of this study was to identify the dominant organisms and pathways involved in hydrogenotrophy in the kangaroo forestomach, with the broader aim of understanding how these processes are able to predominate over methanogenesis. Stable isotope analysis of fermentation end products and RNA stable isotope probing (RNA-SIP) were used to investigate the organisms and biochemical pathways involved in the metabolism of hydrogen and carbon dioxide in the kangaroo forestomach. Our results clearly demonstrate that the activity of bacterial reductive acetogens is a key factor in the reduced methane output of kangaroos. In in vitro fermentations, the microbial community of the kangaroo foregut produced very little methane, but produced a significantly greater proportion of acetate derived from carbon dioxide than the microbial community of the bovine rumen. A bacterial operational taxonomic unit closely related to the known reductive acetogen Blautia coccoides was found to be associated with carbon dioxide and hydrogen metabolism in the kangaroo foregut. Other bacterial taxa including members of the genera Prevotella, Oscillibacter and Streptococcus that have not previously been reported as containing hydrogenotrophic organisms were also significantly associated with metabolism of hydrogen and carbon dioxide in the kangaroo forestomach.The ISME Journal advance online publication, 13 March 2014; doi:10.1038/ismej.2014.25.

Isolation of hen's egg yolk very low density lipoproteins by DEAE-cellulose chromatography

A procedure has been developed for the isolation of very low density lipoproteins from hen's egg yolk plasma using DEAE-cellulose chromatography. This procedure is rapid and does not require ultracentrifugation and should, therefore, serve as a useful procedure for use in laboratories where this facility does not exist.

Presence of 2-methylthioribosyl-trans-zeatin in Azotobacter vinelandii tRNA.

Hydroxylated cytokinin, 2-methylthio-N6-(4-hydroxy-3-methylbut-2-enyl) adenosine, was found in the tRNA of Azotobacter vinelandii. This cytokinin had the trans configuration, unlike the cis configuration reported for that from other bacteria. Culture-condition-dependent changes in the content of this thiocytokinin and a few other thionucleosides in the tRNA of this bacterium have been observed.

N-Carbamoyl-DL-aspartic acid

CsHaN205, PL a = 6.438 (2), b = 7.486 (3), c = 8.048 (4)A, a = 72.2(1), fl = 80.8(1), y = 76.4 (1) °, D m = 1.65 (1) (flotation), D c = 1.64 Mg m -3, Z = 2. Final R = 0.095 for 1205 observed reflections. The molecule assumes the sterically least favourable conformation with the side chain carboxyl group staggered between the a-carboxyl group and the N atom attached to C '~. The ureido group takes part in two specific interactions involving two nearly parallel hydrogen bonds in one and two convergent hydrogen bonds in the other.

Effect of vitamin A deprivation on the cholesterol side-chain cleavage enzyme activity of testes and ovaries of rats (Short Communication)

The cholesterol side-chain cleavage enzyme activity is decreased considerably at the mild stage of vitamin A deficiency in rat testes and ovaries and the decrease in activity becomes more pronounced with progress of deficiency. Supplementation of the deficient rats with retinyl acetate, but not retinoic acid, restores the enzyme activity to normal values. The cholesterol side-chain cleavage enzyme of adrenals is not affected by any of the above treatments.

Effect of LHRH agonists and antagonists in male and female bonnet monkeys (Macaca Radiata)

A few analogues of LHRH have been tested in the adult bonnet monkeys using change in serum testosterone following LHRH injection as a parameter of response to LHRH. Of the four analogues tested in male monkeys, Buserelin was found to be the most potent one in increasing serum testosterone levels. Injection of the LHRH antagonist at 1600 h resulted in the abolition of the characteristic nocturnal surge of testosterone observed in adult bonnet monkeys maintained under regulated light conditions. Following administration of LHRH a/s during early pregnancy, serum chorionic gonadotropin levels decreased though the course of pregnancy was not affected. These results suggest that bonnet monkey can be successfully employed to test LHRH analogues.