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BACKGROUND: Vitamin is a well-known co-factor for many metabolic processes and its roles in fertility and follicular growth have been studied. Vitamin supplementation is frequently achieved by daily ingestion in the form of a complex capsule. However, the role of single and complex vitamins in in vitro maturation of murine follicles is not fully elucidated. METHODS: In this study, we evaluated the effects of two forms of vitamins. Pure L-ascorbic acid, and multi-vitamin (vitamin C+vitamin B complex) was treated at two different concentrations (50 and 100 µg/ml), to pre-puberty murine follicles during in vitro maturation. To determine the specific stage of growth that is affected by treatment with vitamins, the vitamins were treated from day 0, 4, 9, and 13. Growth of each follicle was assessed by measuring diameters of whole expanded area and of the granulosa cells. Expression of follicular and oocyte growth-related genes and the effect of vitamin on the viability of follicles was assessed using senescence associated β-galactosidase staining. RESULTS: Treatment with vitamins promoted the in vitro growth of murine follicles and the upregulated the expression of granulosa cell- and oocyte-specific genes such as BMP15, Fsh receptor, and GDF9. The proliferation of the granulosa cells was enhanced by the treatment of vitamin. Fifty µg/ml concentration vitamin showed greater effects compared to higher concentration. The viability of in vitro grown follicles was also significantly improved in vitamin-treated follicles. The effects of single L-ascorbic acid and complex vitamin were not significantly different to those of day 4 and day 9 follicles. Vitamins promoted murine follicle development in vitro with different effects on specific growth stage. CONCLUSION: Supplementation of vitamins during in vitro maturation of murine follicles is an efficient strategy for in vitro expansion of follicular cells. These results could be customized to the sophisticated culture of follicles retrieved from aged or cancer-survived female that contain smaller number of follicles with reduced potential to develop into mature follicles.
Subject(s)
Female , Humans , Aging , Ascorbic Acid , Eating , Fertility , Granulosa Cells , In Vitro Techniques , Metabolism , Oocytes , Ovarian Follicle , Receptors, FSH , VitaminsABSTRACT
Aim: In the present work, we have studied the effect of L ascorbic acid (LAA) on the regeneration of plants from different families cultured in vitro. Study Design: Plants belonging to three different families are cultured in Murashige and Skoog (MS) medium with and without 1μg/ml L ascorbic acid (LAA), in the absence of any other growth regulators. Thus the study brings out the effect of LAA on plant regeneration. In addition regeneration capacity of LAA involving other growth regulators was also studied. Place and Duration of Study: Department of Biotechnology, Mount Carmel College, Bangalore, India, between August 2012-August 2013. Methodology: The work was conducted on Centella asiatica, Santalum album, and Trigonella foenumgraecum. C. asiatica and T. foenumgraecum are herbaceous whereas S. album is a tropical woody plant. Stem explants of C. asiatica and S. album and the seed explants of T. foenumgraecum were used for the in vitro culture and chlorophyll content in thus obtained leaflets was measured. Further, growth related parameters such as shoot/root length, leaf areas were measured. Results: LAA aided the shoot regeneration in all the three plants cultured in vitro. In C. asiatica and T. foenumgraecum it resulted in the regeneration of plantlets with shoots and roots, however in the case of S. album only shoot regeneration occurred. Chlorophyll content was found to be higher in the in vitro plants grown in the presence of LAA. Shoot/root lengths and area of leaves were more in LAA grown plants as compared to control plants. Conclusion: In vitro culture of stem explants of C. asiatica and seed explants of T. foenumgraecum revealed that supplementing LAA aided in the whole plant regeneration, whereas in the case of S. album supplementing LAA only resulted in the shoot regeneration, but no root formation. Shoot/root lengths, area of leaves and chlorophyll content was found to be higher in the in vitro grown plants with LAA as compared to those grown without LAA, suggesting that LAA is mitigating the function of both auxin and cytokinin. Enhanced chlorophyll production in in-vitro grown plants with LAA is suggestive of involvement of LAA in chlorophyll biosynthesis/protection from degradation and hence the regeneration. Through our results, we show that using LAA in the culture medium can result in regeneration of whole plants. The effect was observed in plants belonging to different families indicating LAA could be used as a general growth enhancer and adding LAA would be beneficial in the regeneration of whole plants.
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Background Melasma is a hypermelanosis which is difficult to treat. There are several treatment options for melasma and one of them is topical therapy using 10% L-Ascorbic acid and 10% Zinc sulfate. Aim To compare the efficacy and side effects of 8 weeks 10% L-Ascorbic acid solution with 10% Zinc sulfate on melasma. Methods This is an observational study with cross sectional design and single-blind, comparing the left and right side of the faces sequentially (right-left comparison study) with each treatment 10% L-Ascorbic acid and 10% Zinc sulfate applied at night. In the morning and afternoon patients uses sunscreen SPF 30. Only new patients with melasma seen at Dermatology Polyclinic Dr M Djamil Hospital Padang from March 2012 to May 2012 were included in this study. Results 20 melasma patients were studied. Their ages range from 25-54 years. 12 (60%) had combination triggering factors. All patients had epidermal type of melasma with 65% located over the centrofacial and 35% on the malar zones. After 2 months of treatment there was significant improvement of melasma treated with 10% Zinc sulfate and 10% L-Ascorbic acid with a P value of <0.05. Minimal side effects were found with Zinc sulfate. Conclusion Improvement of melasma was noted with both topical 10% L-Ascorbic acid and 10% Zinc sulfate but minimal side effects were noted with the use of 10% Zinc sulfate.
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Background: Deficiency of vitamin C (L-ascorbic acid; AA) may induce renal glomular dysfunction in diabetes. Few data are available for the role of continuous upplementation of AA on glomerular dysfunction and pathologyinduced during diabetes. Objective: To investigate long-term effects of AA supplementation on glomerular changes in streptozotocin (STZ)-induced diabetic rats. Methods: Diabetes was induced in Sprague-Dawley rats (180-220 g) by injection of STZ (55 mg/kg bw, iv). The rats were divided into controls (CON), AA-supplemented controls (CON-AA), diabetic (STZ) and AA-supplemented diabetic rats (STZ-AA). AA (1 g/L) was continuously supplemented to the rats for 4, 8, 16 and 24 weeks. The glomerular filtration rate (GFR), effective renal plasma flow (ERPF), renal vascular resistance (RVR) malondialdehyde (MDA) and transforming growth factor-β1 (TGF-β1) levels were measured in the renal cortex. Glomerular morphology was examined histologically. Renal hypertrophic index was calculated using kidney-to-body weight ratio (KW/BW). Results: Decreases in GFR and ERPF were ameliorated at week 16 and deteriorated at week 24 after AA supplementation in STZ-AA rats. High blood glucose concentration was attenuated only at week 16. MAD and TGF-β1 levels in renal cortex decreased significantly in STZ-AA rats at week 16 but not at week 24. The number of abnormal glomeruli and KW/BW decreased significantly at week 16 in STZ-AA rats. Conclusion: Long-term supplementation of AA may ameliorate the glomerular changes induced by diabetes.
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GDP-mannose-3', 5' -epimerase (GME), which converts GDP-mannose into GDP-L-galactose, is essential for the biosynthesis of L-ascorbic acid in higher plants. The molecular characterization of two GME genes from rice has been reported. Firstly,both cDNAs were isolated from the rice mature leaves using RT-PCR technique. By comparing their sequences with homologues from other plants, it was found that GME genes were highly conserved among plant species, though phylogenetic study showed that all known GMEs could be divided into two distinct groups corresponding to monocots and dicots. Secondly, the genomic organization of rice OsGME genes was investigated, and a similarity of splice patterns was revealed. Finally, the expression patterns of the two cDNAs have been studied in various tissues and under different stress conditions by semiquantitative RT-PCR assay. The results showed that the OsGME1 transcript was up-regulated in response to cold stress, and gibberellin might regulate L-ascorbic acid levels by affecting transcription of both OsGME genes.
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Os autores estabeleceram método polarográfico para determinar ácido Lsascórbico na presença de polissorbato eo ou 80 em produtos de panificação, devido à imprecisão na sua determinação pelos métodos usuais. A oxidação anódica do ácido ascórbíco foi realizada em presença da solução-tampão de ace ta to, p H 4,6. Pelo método proposto foram estudadas interferências de alguns aditivos permitidos nas formulações para panificação e também da substância oxidante, o bromato de potássio (AU).
Subject(s)
Polarography , Polysorbates , Ascorbic Acid , BreadABSTRACT
Pretreatment of male Wistar rats with L-ascorbic acid results in a decrease in the in vivo covalent binding of benzo(a)pyrene to hepatic nuclear DNA. In vitro formation of this adduct is also found to be low in liver slices and in liver nuclei of pretreated rats. No inhibition of the adduct formation is, however, observed when benzo (a) pyrene and exogenous DNA are incubated with liver microsomes isolated from ascorbic acid treated rats.It appears that the presence of ascorbate in the cellular or subcellular environment is essential for its inhibitory action.