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Use of high-precision gas isotope ratio mass spectrometry to determine natural abundance 13C in lutein isolated from C3 and C4 plant sources.

A method was developed for high-precision stable carbon isotope ratio analysis of lutein isolated from a C3 (marigold flower) and a C4 (corn gluten meal) plant source using gas chromatography-combustion interfaced isotope ratio mass spectrometry. The natural abundance of 13C (expressed as delta 13C versus the international standard, Pee Dee Belemnite, in per mil units, denoted /1000) in lutein isolated from marigold flower and corn gluten meal was determined to be -29.90 +/- 0.20/1000 and -19.77 +/- 0.27/1000 (mean +/- S.D.), respectively. The high precision of gas isotope ratio mass spectrometry is potentially applicable to detect differences of isotopic composition of lutein in the blood, tissues, or excreta of animal models or humans that result from differences in the natural abundance of 13C in C3 and C4 plant foods.

Carbon Isotopes↗

Effects of external radiation therapy for cancer of the prostate on the serum concentrations of testosterone, follicle-stimulating hormone, luteinizing hormone and prolactin.

Testosterone, luteinizing hormone, follicle-stimulating hormone and prolactin were analyzed in serum from 31 patients with carcinoma of the prostate treated primarily with megavoltage radiation therapy. The total tumor dose varied between 58 and 71 gray (mean 63.5 gray). Absorbed doses to the testes were measured at approximately 1 to more than 10 gray. We investigated retrospectively 17 patients 3 to 60 months (mean 20 months) after therapy and found significantly lower serum testosterone concentrations and significantly higher luteinizing and follicle-stimulating hormone concentrations than in age-matched controls. Of the patients 14 were followed before and after radiation treatment. Testosterone concentrations were reduced significantly 1 week as well as 3 months after treatment but pre-treatment values were found on analysis 6 and 12 months after treatment. The values for luteinizing and follicle-stimulating hormones were significantly higher 3, 6 and 12 months after radiation treatment compared to pre-treatment values. The follicle-stimulating hormone value already increased after 1 week. The greatest observed testosterone alteration occurred 1 week after treatment in patients who received more than 10 gray over the gonads. The use of lead shields protecting the testes reduced the dose absorbed to the gonads by approximately 50 per cent.

Aged↗

Profiling of urinary testosterone and luteinizing hormone in exercise-stressed male athletes, using gas chromatography-mass spectrometry and enzyme immunoassay techniques.

Knowledge of the effects of episodic or short-term exercise-stress on endogenous testosterone and luteinizing hormone levels still remains fragmentary and inconclusive. In this study, an approach based on the absolute concentrations of urinary total testosterone (T), luteinizing hormone (LH) and the T/LH concentration ratios, was used to profile short-term exercise-stress responses in healthy drug-free male athletes. Testosterone and luteinizing hormone concentrations were measured using gas chromatography-mass spectrometry (GC-MS) and microparticle enzyme immunoassay (MEIA) techniques, respectively. Stress profiles derived from exercise-stress at VO2max, 68.1% VO2max and 51.6% VO2max were plotted using the concentrations of T, LH and the ratios of T/LH found under non-stressed and stressed conditions. Significant changes in LH concentrations (p < 0.005) and T/LH ratios (p < 0.005) levels were observed between the pre-stress and post-exercise conditions during acute exercise-stress at VO2max but the T concentration did not show any marked change relative to the non-stressed condition. Whilst exercise-stress appeared to reduce the change in T concentrations between the pre- and post-exercise states compared to that in the non-stressed control condition, the change in LH concentrations showed a moderate increase at submaximal oxygen uptake values. The stress profiles derived from this study facilitated an assessment of the relationship between the endogenous T, LH and T/LH ratio stress-responses over a short period of applied exercise-stress.

Gas Chromatography-Mass Spectrometry↗

Extended coasting duration exerts a negative impact on IVF cycle outcome due to premature luteinization.

Coasting, or withholding gonadotrophin administration while maintaining gonadotrophin-releasing hormone analogue until oestradiol drops to a safe concentration, is an alternative approach to prevent ovarian hyperstimulation syndrome (OHSS) in high responder patients. However, the length of this procedure has not been precisely studied. This paper is a retrospective study of 132 patients who showed a high response (oestradiol > 4500 pg/ml and/or more than 20 follicles > 17 mm) to ovarian stimulation and were coasted due to their high risk of developing OHSS, and evaluated the impact of the duration of coasting on IVF cycle outcome. Additionally, serum LH and progesterone concentrations were studied to investigate whether premature luteinization was present in these cycles and whether it might be related to coasting duration. A significant decrease in implantation rate was found when coasting was required for more than 4 days, together with a trend towards a higher cancellation rate. Premature luteinization was significantly elevated in women undergoing coasting compared with control women (34 versus 15.6%, P < 0.05). In the majority of patients who showed premature luteinization, coasting lasted >/=3 days. To conclude, prolonged coasting may affect the endometrium, anticipating the implantation window. These data may explain why some women undergoing extended coasting show a lower implantation rate compared with controls.

Adult↗

Solubility of purified lutein diesters obtained from Tagetes erecta in supercritical CO(2) and the effect of solvent modifiers.

Purified lutein diesters deposited on commercial nonporous glass beads were solubilized in supercritical CO(2) in a computerized batch extractor, and their solubilities were compared to their solubilities in hexane. Densities of 0.7, 0.8, and 0.9 g/mL were evaluated without modifiers. Both pressure and temperature increased solubility, although temperatures >50 degrees C promoted carotenoid loss as determined by mass balance. Solubility was enhanced by the use of modifiers and was related to their log P. Chloroform (log P = 2) increased 2.8 times the amount of solubilized lutein diesters compared to pure CO(2) at the same extraction conditions (0.9 g/mL and 40 degrees C) to yield 65% of the amount extracted with hexane. Supercritical CO(2) extraction of lutein diesters could represent a cleaner technology as compared to the current industrial use of hexane with important ecological and health-related implications.

Carbon Dioxide↗

Determination of beta-carotene and lutein available from green leafy vegetables by an in vitro digestion and colonic fermentation method.

Green leafy vegetables (Spinacea oleracea, Cnidoscolus aconitifolius, and Solanum americanum) contain a high amount of beta-carotene (27-52 mg/100 g of dry sample) and lutein (140-193 mg/100 g of dry sample). The amount of beta-carotene and lutein released from the food matrix by the action of digestive enzymes ranged from 22 to 67% and from 27 to 77%, respectively. There was a significant correlation between the enzymatic release of carotenoids (lutein + beta-carotene) and the content of Klason lignin, nonstarch polysaccharides, and resistant protein. The carotenoids released by the in vitro colonic fermentation ranged from 2 to 11%, and part of them (0.251-4.03 mg/100 g of original dry sample) remained intact in the fermentation media and could be potentially absorbed in the colon. A significant part of carotenoids seems to be unavailable in the intestinal tract (16% in S. oleracea to 58% in C. aconitifolius).

Biological Availability↗

Bioaccessibility of beta-carotene, lutein, and lycopene from fruits and vegetables.

Epidemiological studies have consistently demonstrated that there is an association between carotenoid-rich food intakes with a low incidence in chronic diseases. Nevertheless, there is not an association between the intake of total dietary carotenoids and chronic health incidence in the European population, probably because of different carotenoid food sources and bioavailability. The objective of this study was to evaluate the small and large intestine bioaccessibilities of major dietary carotenoids from fruits and vegetables in a common diet. A bioaccessibility model that includes enzymatic digestion and in vitro colonic fermentation was employed. Lutein presented greater small intestine bioaccessibility (79%) than beta-carotene (27%) or lycopene (40%). With regard to large intestine bioaccessibility, similar amounts of lycopene and beta-carotene were released from the food matrix (57%), whereas small amounts of lutein (17%) were released. These results suggest that 91% of the beta-carotene, lutein, and lycopene contained in fruits and vegetables is available in the gut during the entire digestion process. Colonic fermentation is shown to be important for carotenoid availability in the gut.

Animals↗

Non-nutritive bioactive constituents of plants: lycopene, lutein and zeaxanthin.

Lycopene, lutein, and zeaxanthin are major carotenoids in human blood and tissues but unlike beta-carotene do not contribute to vitamin A supply. These carotenoids are efficient antioxidants quenching singlet molecular oxygen which is formed in photooxidative processes and thus may contribute to the prevention of light-exposed tissue, skin and eyes, from light-induced damage. Increasing lycopene intake by daily consumption of tomato paste over a period of ten weeks provides protection against erythema formation following UV-irradiation. Lycopene and other carotenoids may be used as oral sun protectants and contribute to the maintenance of skin health. The yellow color of the macula lutea is due to the presence of the carotenoid pigments lutein and zeaxanthin. These macula carotenoids are suggested to play a role in protection against light-dependent damage. Filtering of blue light and scavenging of reactive intermediates generated in photooxidation are considered to be the underlying protective mechanisms. Epidemiological studies provide evidence that an increased consumption of lutein is associated with a lowered risk for age-related macular degeneration, a disease with increasing incidence in the elderly.

Aging↗

Evolution of the genes for the beta subunits of human chorionic gonadotropin and luteinizing hormone.

Nucleotide sequence comparisons of the single gene for the human luteinizing hormone gene beta subunit with two of the seven genes for the human chorionic gonadotropin beta subunit suggest that the beta human chorionic gonadotropin genes have evolved from an ancestral beta luteinizing hormone gene by a series of selected changes with very little neutral drift. Moreover, the 24 amino acid carboxy-terminal extension of the human chorionic gonadotropin beta subunit appears to have arisen by a single base deletion that incorporated the 3'-untranslated region of the ancestral beta luteinizing hormone gene into the coding region.

Base Sequence↗

Influence of thyroxine and luteinizing hormone on some enzymes concerned with lopogenesis in adipose tissue, testis and adrenal gland.

The activities of hexokinase, citrate-cleavage enzyme, ;malic enzyme' and NADP-linked isocitrate dehydrogenase have been measured in the adipose tissue, testes and adrenals of normal rats, hypophysectomized rats and hypophysectomized rats treated with either thyroxine or thyroxine plus luteinizing hormone. Hypophysectomy reduced the activity of all four enzymes in all three tissues. Thyroxine alone restored the activity of all four enzymes in adipose tissue towards normal but failed to do so in either testes or adrenals. Thyroxine and luteinizing hormone restored the citrate-cleavage enzyme activity of testes and increased the activity of hexokinase from the low value after hypophysectomy. Neither ;malic enzyme' nor isocitrate dehydrogenase was increased by thyroxine or thyroxine and luteinizing hormone in testes. The differential stimulation of enzyme activity by thyroxine in the different tissues suggests thyroxine as having a special significance in adipose-tissue lipogenesis.

Adipose Tissue↗

The purification of luteinizing-hormone-releasing factor with some observations on its properties.

Bullock median-eminence tissue was used as a source of luteinizing-hormone-releasing factor for small-scale experiments to explore methods for its isolation. The presence of luteinizing-hormone-releasing factor was detected by the ovulation response in rabbits after intrapituitary infusion of the extract. Gel filtration was found to be suitable for the purification of these extracts. The releasing factor appeared to be a basic peptide of molecular weight in the range 1200-2500. On a larger scale, an extract of hypothalamic tissue from sheep was used to establish a multi-stage isolation procedure that resulted in a 200000-fold purification of luteinizing-hormone-releasing factor. After the initial extraction the isolation process consisted of: (1) two cycles of gel filtration; (2) anion-exchange chromatography; (3) gel filtration in a partially organic medium; (4) thin-layer chromatography on cellulose. Stage (3) separated two zones of activity each containing peptides. One of these was purified further by stage (4) to give a preparation that was active at a dose of 6mug. of peptide/animal, although activity diminished seriously during storage. This preparation contained only five or six components, but the small amounts of peptides obtained at this stage of purity were insufficient for full characterization.

Animals↗

The hormone-binding unit of luteinizing hormone receptor.

Rat ovarian luteinizing hormone/human choriogonadotropin binding sites were labelled with 125I-choriogonadotropin in vivo, and the resulting 125I-choriogonadotropin-receptor complexes were solubilized by Triton X-100 and purified by use of antibodies to choriogonadotropin immobilized to agarose. The purified 125I-choriogonadotropin-receptor complex was treated with glutaraldehyde to crosslink radiolabelled hormone to the receptor. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of the crosslinked product revealed a labelled Mr 130 000 major band in addition to the hormone and its alpha-subunit, indicating that a single receptor component was linked to the hormone. Unoccupied binding sites for luteinizing hormone were also solubilized by Triton X-100 from pseudopregnant rat ovaries, and attached to choriogonadotropin-agarose. The agarose gel was washed, and eluted with 0.1 M-sodium acetate, pH 4. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of the pH 4 eluate revealed an Mr 90 000 major band which was abolished when ovaries presaturated with choriogonadotropin were used as starting material. These observations suggest that the hormone-binding component of the luteinizing hormone receptor is a polypeptide of Mr 90 000. This polypeptide was isolated and labelled with Na 125I. The labelled polypeptide showed a single band on sucrose density gradient centrifugation and on gel filtration on agarose.

Animals↗

Isolation and partial characterization of tammar wallaby luteinizing hormone and development of a radioimmunoassay.

Tammar wallaby (Macropus eugenii) luteinizing hormone (LH) was purified from pituitaries collected from wild and captive populations by salt sequential precipitation, ion exchange chromatography and gel filtration. Pituitary tissue (5 g) yielded 1.8 mg of purified wallaby luteinizing hormone (ME-14B), as verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). A heterologous radioimmunoassay has been developed for measurement of LH in plasma of marsupials using a monoclonal antibody raised against bovine LH (518B7). This assay system was able to measure basal LH concentrations in male and female tammars and detected a significant rise in plasma LH in response to oestradiol benzoate in female tammars and luteinizing hormone-releasing hormone (LHRH) in males. Parallel dose-response curves were also obtained from pituitary extracts from four other species of marsupial (brushtail possum, Trichosurus vulpecula; brown antechinus, Antechinus stuartii; kowari, Dasyuroides byrnei; and Eastern pygmy possum, Cercartetus nanus) in this assay, which suggests its usefulness in the measurement of LH in other marsupial species.

Animals↗

Inhibition of prostate tumor growth in two rat models by chronic administration of D-Trp6 analogue of luteinizing hormone-releasing hormone.

We have investigated the effect of the D-Trp6 analogue of luteinizing hormone-releasing hormone (LH-RH), a superactive analogue of LH-RH, on the growth of two different models of prostate tumors in rats. Chronic administration of D-Trp6-LH-RH in a dose of 25 micrograms/day for 14-21 days significantly inhibited the growth of the chemically induced squamous cell carcinoma 11095 in Fisher 344 male rats. The weights of the ventral prostate and testes were also significantly reduced by treatment with this analogue. After 21 days of treatment, the animals no longer showed increases in serum luteinizing hormone and follicle-stimulating hormone levels in response to D-Trp6-LH-RH. Treatment of male Copenhagen F-1 rats bearing the Dunning 3327 prostate adenocarcinoma with 25 micrograms of D-Trp6-LH-RH per day for 42 days decreased the weights of both the ventral prostate and testes but had no effect on the weight of the anterior pituitary gland. The percentage increase in tumor volume was decreased to one-third and the actual tumor weight was decreased by 58% compared to untreated controls. The tumor doubling time was more than 4 times longer in rats receiving D-Trp6-LH-RH than in controls. Serum levels of luteinizing hormone and follicle-stimulating hormone were significantly decreased in rats receiving this analogue. In both Fisher 344 and Copenhagen F-1 rats, serum prolactin and testosterone levels were significantly decreased after treatment with D-Trp6-LH-RH, whereas progesterone levels were increased.

Animals↗

Prostate carcinoma tumor size in rats decreases after administration of antagonists of luteinizing hormone-releasing hormone.

The effects of two potent antagonistic analogues of luteinizing hormone-releasing hormone (LH-RH) on the growth of two different models of rat prostate tumors have been investigated. Chronic administration of [NAc-p-F-DPhe1,p-Cl-DPhe2,DTrp3.6,D-Ala10]LH-RH (antagonist I) at 50 micrograms/day for 21 days significantly inhibited the growth of the chemically induced squamous cell carcinoma 11095 in Fisher 344 male rats. The weights of the pituitary, ventral prostate, and testes were not significantly altered. After 21 days of treatment with this analogue serum luteinizing hormone (lutropin), follicle-stimulating hormone, and testosterone levels were markedly decreased. When male Copenhagen F-1 rats bearing the Dunning 3327H prostate adenocarcinoma were injected with antagonist I at 50 micrograms/day for 6 weeks or with [NAc-p-Cl-DPhe1,2,DTrp3,DPhe6,DAla10]LH-RH (antagonist II) at 50 micrograms/day for 17 days, the percentage increase in tumor volume was decreased to half or less and the actual tumor volume was diminished 34-96% compared to controls. Tumor weight was decreased 30% and 89% after antagonist I and, respectively, compared to untreated controls. The tumor doubling time was 3- to 4-fold longer in rats receiving the inhibitory analogues than in the controls. Treatment with antagonist II decreased the weight of the whole prostate, but neither antagonist changed the weight of testes, anterior pituitary gland, or adrenals. Serum luteinizing hormone, follicle-stimulating hormone, and testosterone levels in Copenhagen F-1 rats bearing Dunning tumors were significantly decreased after treatment with the inhibitory analogues, but progesterone levels were increased. The inhibitory effects of these antagonistic analogues on rat prostate tumors suggest that these compound might be considered in the development of new types of therapy for prostate carcinoma and other endocrine-dependent neoplasias.

Animals↗

Highly potent antagonists of luteinizing hormone-releasing hormone free of edematogenic effects.

To eliminate the undesirable edematogenic effect of the luteinizing hormone-releasing hormone (LH-RH) antagonists containing basic D amino acids at position 6, exemplified by [Ac-D-Phe(pCl)1,2,D-Trp3,D-Arg6,D-Ala10]LH-RH [Phe(pCl) indicates 4-chlorophenylalanine], analogs with D-ureidoalkyl amino acids such as D-citrulline (D-Cit) or D-homocitrulline (D-Hci) at position 6 were synthesized and tested in several systems in vitro and in vivo. HPLC analysis revealed that the overall hydrophobicity of the D-Cit/D-Hci6 analogs was similar to that of the basic D-Arg6 antagonists. In vitro, most of the analogs completely inhibited LH-RH-mediated luteinizing hormone release in perfused rat pituitary cell systems at an antagonist to LH-RH molar ratio of 5:1. In vivo, the most active peptides, [Ac-D-Nal(2)1,D-Phe(pCl)2,D-Trp3,D-Cit6,D-Ala10]LH-RH [Nal(2) indicates 3-(2-naphthyl)alanine] and its D-Hci6 analog, caused 100% inhibition of ovulation in cycling rats in doses of 3 micrograms and suppressed the luteinizing hormone level in ovariectomized female rats for 47 hr when administered at doses of 25 micrograms. Characteristically, these peptides did not exert any edematogenic effects even at 1.5 mg/kg. These properties of the D-Cit/D-Hci6 antagonists may make them useful clinically.

Anaphylaxis↗

The correlation between the intake of lutein, lycopene and beta-carotene from vegetables and fruits, and blood plasma concentrations in a group of women aged 50-65 years in the UK.

The correlations between the mean of 4 d weighed intakes of lutein, lycopene and beta-carotene and mean plasma concentrations during each of the four seasons were lutein r 0.64, lycopene r 0.47 and beta-carotene r 0.45. Intake was not significantly correlated with plasma concentrations during every seasonal time-point. There was a significantly higher intake of lutein during the spring compared with summer and autumn, lycopene intake was significantly higher during the summer and autumn and there were no significant seasonal differences in beta-carotene intake. There were, however, significant seasonal differences in plasma carotenoid concentrations, the highest levels occurring between May and October. There were large inter- and intra-individual variations in intake and plasma concentrations of carotenoids. BMI was inversely correlated with plasma beta-carotene (r-0.41). The findings suggest that plasma carotenoid concentrations are indicative of dietary intake, but the large intra-individual variation in plasma concentrations indicates that any assessment of longer-term status from data at any one time-point should be treated with caution.

Aged↗

No significant effects of lutein, lycopene or beta-carotene supplementation on biological markers of oxidative stress and LDL oxidizability in healthy adult subjects.

OBJECTIVE: The objective of this study was to determine the effect of individual carotenoid supplementation on biochemical indices of oxidative status in apparently healthy adult males. METHODS: The study was a placebo controlled single blind study. Healthy male volunteers (n= 175) were assigned to four groups. They received daily supplements of beta-carotene (15 mg), lutein (15 mg), lycopene (15 mg) and placebo for three months. The effects of the supplementation on antioxidant status were monitored by plasma carotenoid, vitamin C and A levels, glutathione (GSH and GSSG) concentrations, protein SH groups. erythrocyte antioxidant enzyme activities (Cu-Zn SOD, Se-GSH-Px) and susceptibility of LDL to copper-induced oxidation. RESULTS: beta-carotene, lycopene and lutein supplementation led to significant plasma and LDL increases in each of these carotenoids, without modifications of other carotenoid levels in plasma or in LDL. The supplementation failed to enhance the resistance of LDL to oxidation or to modify the LDL polyunsaturated/ saturated fatty acid ratio. Vitamin C, GSH, protein SH groups and antioxidant metalloenzyme activities were also unchanged. CONCLUSION: We did not observe beneficial or adverse effects of lutein, lycopene or beta-carotene supplementation on biomarkers of oxidative stress. In apparently healthy subjects, carotenoid supplementation does not lead to significantly measurable improvement in antioxidant defenses.

Adult↗