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1.
Mycologia ; 116(3): 464-474, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38489159

RESUMO

Tremella fuciformis Berk. (TF), or the white jelly mushroom, is well known for its myriad of pharmacological properties, such as immunomodulatory, anti-inflammatory, antidiabetic, antitumor, and antioxidant activities, and hypocholesterolemic and hepatoprotective effects that boost human health. Most of the studies of TF are concentrated on its polysaccharide (glucuronoxylomannan) composition, which is responsible for its pharmacological as well as rheological properties. It is well established that mushrooms are a great source of dietary vitamin D due to the presence of ergosterol in their cell membrane. There is a lack of published data on TF as a source of vitamin D2. Therefore, this study aimed to evaluate the vitamin D2 composition of the fruiting bodies of TF using triple quadrupole liquid chromatography-mass spectrometry (LC-MS/QQQ). The results showed highest vitamin D2 content (292.02 µg/g dry weight) in the sample irradiated with ultraviolet B (UVB; 310 nm) for 180 min as compared with the control group (52.47 µg/g dry weight) (P ≤ 0.001). The results showed higher accumulation potential of vitamin D2 in TF as compared with published data available for other extensively studied culinary mushrooms, such as Agaricus bisporus, Lentinula edodes, Pleurotus ostreatus, Cordiceps militaris, and Calocybe indica. Moreover, the impact of UV treatment on antioxidant capacities and total polyphenol content of TF was also studied. The accumulation potential of vitamin D in TF reveals a novel commercial source for this nutrient.


Assuntos
Antioxidantes , Ergocalciferóis , Polifenóis , Ergocalciferóis/metabolismo , Ergocalciferóis/análise , Polifenóis/metabolismo , Polifenóis/análise , Antioxidantes/metabolismo , Antioxidantes/análise , Cromatografia Líquida , Basidiomycota/metabolismo , Basidiomycota/química , Carpóforos/química , Carpóforos/metabolismo , Agaricales/química , Agaricales/metabolismo , Espectrometria de Massas , Espectrometria de Massas em Tandem , Espectrometria de Massa com Cromatografia Líquida
2.
Int J Mol Sci ; 24(5)2023 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-36902073

RESUMO

Vitamin D is a fat-soluble secosteroid that exists in two forms: vitamin D2 and vitamin D3 [...].


Assuntos
Proteína de Ligação a Vitamina D , Vitamina D , Colecalciferol/metabolismo , Ergocalciferóis/metabolismo , Vitamina D/metabolismo , Proteína de Ligação a Vitamina D/metabolismo
3.
Exp Biol Med (Maywood) ; 248(2): 186-192, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36373746

RESUMO

The vascular endothelium is one of the main targets of oxidative stress which plays an important role in the pathophysiology of vascular damage. Recent studies show that vitamin D can positively regulate endothelial functions in various chronic diseases and in cases of increased oxidative stress. In our study, we investigated the restorative and protective potentials of paricalcitol which is frequently used in patients with chronic renal failure, a vitamin D analogue, in human umbilical vein endothelial cells (HUVEC) before and after H2O2-induced oxidative stress. Paricalcitol treatment after the oxidative stress induced by H2O2 increased cell viability in endothelial cells depending on the dose that was used. While paricalcitol (500 nM) decreased caspase-3 activity and mitochondrial membrane potential loss, it increased nitric oxide (NO) production and reduced glutathione (GSH) levels. Paricalcitol treatment before oxidative stress increased cell viability. It increased NO production and mitochondrial membrane potential while significantly reducing caspase-3 activity. While paricalcitol caused a significant inhibition of protein disulfide isomerase (PDI) reductase activity in healthy endothelial cells, it did not cause a significant change on the PDI reductase activity under oxidative stress conditions. Present study showed that paricalcitol has restorative and protective effects on endothelial cells against oxidative injury, but these effects occur at high concentrations of paricalcitol.


Assuntos
Apoptose , Peróxido de Hidrogênio , Humanos , Peróxido de Hidrogênio/toxicidade , Caspase 3/metabolismo , Caspase 3/farmacologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Estresse Oxidativo , Ergocalciferóis/farmacologia , Ergocalciferóis/metabolismo , Óxido Nítrico/metabolismo , Oxirredutases/metabolismo , Oxirredutases/farmacologia , Espécies Reativas de Oxigênio/metabolismo
4.
Biotechnol Lett ; 44(10): 1217-1230, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36057882

RESUMO

Ergosterol as a primary metabolite and precursor of vitamin D2, is the most plentiful mycosterols in fungal cell membrane. Process optimization to increase the yield and productivity of biological products is a topic of interest. Ultrasonic waves have many applications in biotechnology, like cell disruption, and enhancement of primary and secondary metabolites production. This study disclosed an optimal condition for ultrasound-assisted production (UAP) of ergosterol from Penicillium brevicompactum MUCL 19,011 using L9 Taguchi statistical method. The intensity (IS), time of sonication (TS), treatment frequency (TF), and number of days of treatment (DT) were allocated to study the effects of ultrasound on ergosterol production. The results were analyzed using Minitab version 19. The maximum ergosterol, 11 mg/g cell dry weight (CDW), was produced on the tenth day while all factors were at a low level. The days of treatment with a contribution of 45.48% was the most significant factor for ergosterol production. For the first time, this study revealed the positive effect of ultrasound on the production of ergosterol. Ergosterol production increased 73% (4.63 mg/g CDW) after process optimization. Finally, a mathematical model of ultrasound factors with a regression coefficient of R2 = 0.978 was obtained for the ergosterol production during ultrasound treatment.


Assuntos
Produtos Biológicos , Penicillium , Produtos Biológicos/metabolismo , Ergocalciferóis/metabolismo , Ergosterol/metabolismo , Penicillium/genética , Penicillium/metabolismo
5.
Chembiochem ; 22(13): 2266-2274, 2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-33647186

RESUMO

The active vitamin D metabolites 25-OH-D and 1α,25-(OH)2 -D play an essential role in controlling several cellular processes in the human body and are potentially effective in the treatment of several diseases, such as autoimmune diseases, cardiovascular diseases and cancer. The microbial synthesis of vitamin D2 (VD2 ) and vitamin D3 (VD3 ) metabolites has emerged as a suitable alternative to established complex chemical syntheses. In this study, a novel strain, Kutzneria albida, with the ability to form 25-OH-D2 and 25-OH-D3 was identified. To further improve the conversion of the poorly soluble substrates, several solubilizers were tested. 100-fold higher product concentrations of 25-OH-D3 and tenfold higher concentrations of 25-OH-D2 after addition of 5 % (w/v) 2-hydroxypropyl ß-cyclodextrin (2-HPßCD) were reached. Besides the single-hydroxylation products, the human double-hydroxylation products 1,25-(OH)2 -D2 and 1,25-(OH)2 -D3 and various other potential single- and double-hydroxylation products were detected. Thus, K. albida represents a promising strain for the biotechnological production of VD2 and VD3 metabolites.


Assuntos
Actinobacteria/metabolismo , Colecalciferol/metabolismo , Ergocalciferóis/metabolismo , Colecalciferol/química , Ergocalciferóis/química , Hidroxilação , Estrutura Molecular
6.
Int J Mol Sci ; 23(1)2021 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-35008598

RESUMO

BACKGROUND: Ovarian cancer (OC) is one of the most lethal cancers in women. The active form of vitamin D3, 1,25-dihydroxyvitamin D3 (1,25D3, calcitriol) has anticancer activity in several cancers, including ovarian cancer, but the required pharmacological doses may cause hypercalcemia. We hypothesized that newly developed, low calcemic, vitamin D analogs (an1,25Ds) may be used as anticancer agents instead of calcitriol in ovarian cancer cells. METHODS: We used two patient-derived high-grade serous ovarian cancer (HGSOC) cell lines with low (13781) and high (14433) mRNA expression levels of the gene encoding 1,25-dihydroxyvitamin D3 24-hydroxylase CYP24A1, one of the main target genes of calcitriol. We tested the effect of calcitriol and four structurally related series of an1,25Ds (PRI-1906, PRI-1907, PRI-5201, PRI-5202) on cell number, viability, the expression of CYP24A1, and the vitamin D receptor (VDR). RESULTS: CYP24A1 mRNA expression increased in a concentration-dependent manner after treatment with all compounds. In both cell lines, after 4 h, PRI-5202 was the most potent analog (in 13781 cells: EC50 = 2.98 ± 1.10 nmol/L, in 14433 cells: EC50 = 0.92 ± 0.20 nmol/L), while PRI-1907 was the least active one (in 13781 cells: EC50 = n/d, in 14433 cells: EC50 = n/d). This difference among the analogs disappeared after 5 days of treatment. The 13781 cells were more sensitive to the an1,25Ds compared with 14433 cells. The an1,25Ds increased nuclear VDR levels and reduced cell viability, but only in the 13781 cell line. CONCLUSIONS: The an1,25Ds had different potencies in the HGSOC cell lines and their efficacy in increasing CYP24A1 expression was cell line- and chemical structure-dependent. Therefore, choosing sensitive cancer cell lines and further optimization of the analogs' structure might lead to new treatment options against ovarian cancer.


Assuntos
Sobrevivência Celular , Neoplasias Ovarianas/tratamento farmacológico , Receptores de Calcitriol/genética , Vitamina D3 24-Hidroxilase/genética , Vitamina D/farmacologia , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Células Cultivadas , Ergocalciferóis/metabolismo , Ergocalciferóis/farmacologia , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Ovarianas/enzimologia , Neoplasias Ovarianas/metabolismo , Vitamina D/análogos & derivados
7.
Int J Mol Sci ; 21(21)2020 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-33105665

RESUMO

Vitamin D is an important component of the endocrine system that controls calcium homeostasis and bone mineralization. Because of the very short half-life of free serum vitamin D it is stabilized and transported to target tissues by being bound to the vitamin D binding protein (VDBP). The most common polymorphisms: rs4588 and rs7041 in the vitamin D binding protein gene may correlate with differences in vitamin D status in the serum. This review presents data that relate to the presence of genetic variants in the VDBP gene in correlation with certain diseases, mostly concerning cancers (breast, prostate, pancreatic, lung, colorectal, basal cell carcinoma cancer and cutaneous melanoma) or other related diseases (thyroid autoimmunity disorders, obesity, diabetes mellitus, bone metabolism, rheumatoid arthritis, ankylosing spondylitis, asthma, chronic obstructive pulmonary disease, tuberculosis and coronary artery diseases).


Assuntos
Neoplasias/genética , Polimorfismo de Nucleotídeo Único , Proteína de Ligação a Vitamina D/genética , Artrite Reumatoide/genética , Colecalciferol/metabolismo , Doença da Artéria Coronariana/genética , Diabetes Mellitus/genética , Ergocalciferóis/metabolismo , Feminino , Predisposição Genética para Doença , Humanos , Masculino , Obesidade/genética , Doença Pulmonar Obstrutiva Crônica/genética , Tuberculose/genética , Proteína de Ligação a Vitamina D/fisiologia
8.
Biochem Biophys Res Commun ; 524(2): 295-300, 2020 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-31987498

RESUMO

Vitamin D2 is a form of vitamin D derived from mushrooms and plants which is structurally modified in the body due to the action of several enzymes. The resulting metabolites represent important compounds with potential bioactive properties. However, they are poorly studied and their availability is mostly limited. In order to identify new enzymes capable of producing vitamin D2 metabolites, we investigated a bacterial P450 monooxygenase, CYP109E1, which was previously shown to be a vitamin D3 hydroxylase. It was found that CYP109E1 catalyzes a vitamin D2 two-step hydroxylation at positions C24 and C25 resulting in the generation of 24(R),25-diOH VD2. Interestingly, the enzyme showed high selectivity towards vitamin D2, whereas it showed an unselective product pattern for the structurally similar vitamin D3. Our docking results for vitamin D2 and D3 revealed favorable hydroxylation positions for both substrates and suggested an explanation for the high selectivity of CYP109E1 towards vitamin D2. In addition, we established a whole-cell biocatalyst expressing CYP109E1 in Bacillus megaterium to produce 24(R),25-diOH VD2 and a production yield of 12.3 ± 1.2 mg/L was obtained after 48 h. To the best of our knowledge, this is the first report on the generation of 24(R),25-diOH VD2 by a microbial biocatalyst allowing a low-cost and eco-friendly production of this pharmaceutically interesting and expensive metabolite from the relatively cheap substrate, VD2.


Assuntos
Bacillus megaterium/metabolismo , Proteínas de Bactérias/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Ergocalciferóis/metabolismo , Bacillus megaterium/enzimologia , Hidroxilação , Simulação de Acoplamento Molecular , Estereoisomerismo , Especificidade por Substrato
9.
Food Chem ; 309: 125738, 2020 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-31706679

RESUMO

In the present study, the impact of ultraviolet (UV)-C treatment and ultrafine grinding on the conversion of ergosterol to vitamin D2, physiochemical properties, and antioxidant properties of shiitake and Jew's ear was assessed. After exposure to UV-C, vitamin D2 contents of both the mushroom samples has increased significantly (p < 0.05). Whereas, ultrafine grinding along with UV-C treatment has a synergistic effect on bioconversion of ergosterol to vitamin D2 and this effect is more prominent in low dose UV-C irradiation groups (2 kJ/m2). Ultrafine grinding significantly (p < 0.05) improved the water holding capacity (WHC), water solubility index (WSI) and polysaccharide dissolution rate (PDR). However, UV-C treatment led to insignificant changes in the physiochemical properties of mushroom samples. A significant improvement was also observed in the antioxidant profiles especially tannin contents of mushrooms followed by the ultrafine grinding and UV-C treatment.


Assuntos
Agaricales/metabolismo , Agaricales/efeitos da radiação , Antioxidantes/metabolismo , Ergocalciferóis/metabolismo , Ergosterol/metabolismo , Cogumelos Shiitake/metabolismo , Cogumelos Shiitake/efeitos da radiação , Agaricales/química , Antioxidantes/química , Biotransformação , Ergocalciferóis/química , Ergosterol/química , Cogumelos Shiitake/química , Raios Ultravioleta
10.
Nutr Hosp ; 36(Spec No3): 3-7, 2019 Aug 27.
Artigo em Espanhol | MEDLINE | ID: mdl-31368328

RESUMO

INTRODUCTION: Vitamin D is an essential nutrient whose deficiency has been associated with the risk of various chronic diseases such as osteoporosis, hypertension, cardiovascular disease, diabetes, some types of cancer and even overweight and obesity. Although vitamin D can be synthesized at the skin from exposure to sunlight, this source is not always sufficient to meet the needs. For example, the use of sunscreen or the low exposition to the sunlight limits the syntheses. In fact, studies have found that at least half of the Spanish population has vitamin D deficits. Therefore, the dietary contribution is fundamental. Although there are different foods fortified in this vitamin, few products are natural source of it, as fatty fish and eggs. However, according to different studies carried out in the Spanish population, there is a low consumption of this food group. In this way, it would be advisable to promote egg consumption among the population, since this food, in addition to having many nutrients, contains a high amount of vitamin D, which contributes to avoid the appearance of deficiencies and the consequences health consequences that this implies.


INTRODUCCIÓN: La vitamina D es un nutriente esencial cuya deficiencia se ha asociado con el riesgo de aparición de diversas enfermedades crónicas, como la osteoporosis, la hipertensión arterial, la enfermedad cardiovascular, la diabetes, algunos tipos de cáncer e incluso el padecimiento de sobrepeso y obesidad. A pesar de que la vitamina D puede sintetizarse a nivel cutáneo a partir de la exposición a la luz solar, esta fuente no es siempre suficiente para cubrir las necesidades debido al uso de cremas de protección solar y a la baja exposición que se produce durante el invierno, o, como en el caso de las personas enfermas, que salen poco a la calle o se exponen poco a la luz del sol. De hecho, estudios han constatado que al menos la mitad de la población española presenta déficit de vitamina D. Por ello, el aporte dietético es fundamental. Aunque existen diferentes alimentos fortificados con esta vitamina, son pocos los productos que son una fuente natural, entre los que se encuentran los pescados grasos y los huevos. Sin embargo, de acuerdo con diferentes estudios realizados en la población española, existe un bajo consumo de este último grupo de alimentos. De esta manera, sería recomendable fomentar el consumo de huevo entre la población, ya que este alimento, además de tener numerosos nutrientes, contiene una cantidad elevada de vitamina D, lo que contribuye a evitar la aparición de deficiencias y las consecuencias negativas para la salud que ello implica.


Assuntos
Colecalciferol/administração & dosagem , Ovos , Ergocalciferóis/administração & dosagem , Deficiência de Vitamina D/dietoterapia , Vitaminas/administração & dosagem , Colecalciferol/biossíntese , Colecalciferol/sangue , Ergocalciferóis/sangue , Ergocalciferóis/metabolismo , Humanos , Política Nutricional , Espanha/epidemiologia , Luz Solar , Vitamina D/biossíntese , Vitamina D/sangue , Deficiência de Vitamina D/complicações , Deficiência de Vitamina D/epidemiologia
11.
Meat Sci ; 143: 60-68, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29715661

RESUMO

This study investigated the effects of synthetic and natural sources of vitamin D biofortification in pig diets on pork vitamin D activity and pork quality. One hundred and twenty pigs (60 male, 60 female) were assigned to one of four dietary treatments for a 55 d feeding period. The dietary treatments were (1)50 µg vitamin D3/kg of feed; (2)50 µg of 25-hydroxvitamin D3/kg of feed (25-OH-D3); (3)50 µg vitamin D2/kg of feed; (4)50 µg vitamin D2-enriched mushrooms/kg of feed (Mushroom D2). The pigs offered the 25-OH-D3 diet exhibited the highest (P < 0.001) serum total 25-hydroxyvitamin D concentration and subsequently exhibited the highest (P < 0.05) Longissimus thoracis (LT) total vitamin D activity. Mushroom D2 and 25-OH-D3 supplementation increased pork antioxidant status. The vitamin D2-enriched mushrooms improved (P < 0.05) pig performance, carcass weight and LT colour. In conclusion, 25-OH-D3 is the most successful source for increasing pork vitamin D activity, while Mushroom D2 may be a new avenue to improve animal performance and pork quality.


Assuntos
Agaricales/química , Fenômenos Fisiológicos da Nutrição Animal , Antioxidantes/administração & dosagem , Calcifediol/administração & dosagem , Qualidade dos Alimentos , Carne/análise , Músculo Esquelético/metabolismo , 25-Hidroxivitamina D 2/sangue , Agaricales/crescimento & desenvolvimento , Agaricales/metabolismo , Animais , Antioxidantes/análise , Antioxidantes/metabolismo , Calcifediol/análise , Calcifediol/sangue , Calcifediol/metabolismo , Colecalciferol/administração & dosagem , Colecalciferol/análise , Colecalciferol/metabolismo , Cruzamentos Genéticos , Ergocalciferóis/administração & dosagem , Ergocalciferóis/análise , Ergocalciferóis/metabolismo , Feminino , Alimentos Fortificados/análise , Humanos , Irlanda , Masculino , Músculo Esquelético/crescimento & desenvolvimento , Valor Nutritivo , Pigmentos Biológicos/análise , Pigmentos Biológicos/biossíntese , Distribuição Aleatória , Sus scrofa , Aumento de Peso
12.
Food Chem ; 256: 18-24, 2018 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-29606435

RESUMO

This study investigates dietary fortification of heifer feeds with cholecalciferol and ergocalciferol sources and effects on beef total vitamin D activity, vitamer, respective 25-hydroxymetabolite contents, and meat quality. Thirty heifers were allocated to one of three dietary treatments [(1) basal diet + 4000 IU of vitamin D3 (Vit D3); (2) basal diet + 4000 IU of vitamin D2 (Vit D2); and (3) basal diet + 4000 IU of vitamin D2-enriched mushrooms (Mushroom D2)] for a 30 day pre-slaughter period. Supplementation of heifer diets with Vit D3 yielded higher (p < 0.001) Longissimus thoracis (LT) total vitamin D activity (by 38-56%; p < 0.05) and serum 25-OH-D concentration (by 20-36%; p < 0.05), compared to that from Vit D2 and Mushroom D2 supplemented animals. Irrespective of vitamin D source, carcass characteristics, sensory and meat quality parameter were unaffected (p > 0.05) by the dietary treatments. In conclusion, vitamin D3 biofortification of cattle diets is the most efficacious way to enhance total beef vitamin D activity.


Assuntos
Agaricales/efeitos da radiação , Colecalciferol/administração & dosagem , Ergocalciferóis/administração & dosagem , Alimentos Fortificados/análise , Carne/análise , Raios Ultravioleta , Agaricales/metabolismo , Animais , Músculos do Dorso/química , Músculos do Dorso/metabolismo , Calcifediol/análise , Calcifediol/sangue , Cálcio/sangue , Bovinos , Colecalciferol/síntese química , Cromatografia Líquida de Alta Pressão , Dieta/veterinária , Ergocalciferóis/metabolismo , Espectrometria de Massas em Tandem
13.
Int J Med Mushrooms ; 19(6): 547-560, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29199564

RESUMO

The caterpillar medicinal mushroom Cordyceps militaris contains many bioactive components, such as adenosine, cordycepin, and polysaccharides. In this study, C. militaris was exposed to 0, 3, 6, or 9 pulses of light irradiation to estimate changes in vitamin D2, bioactive compounds, nonvolatile taste components, and antioxidant properties. In addition, we compared the components and properties of C. militaris mycelia and solid waste medium that had been treated with pulsed light (PL) irradiation. Overall, PL irradiation of C. militaris increased the vitamin D2 content and increased the total amino acid levels 9-48%; the antioxidant properties of the mycelia treated with 0 pulses and of the solid waste medium treated with 3 pulses all exhibited lower half-maximal effective concentrations. Therefore, PL irradiation affected the amounts of bioactive compounds, but the irradiated samples still contained intense umami taste and a sufficient amount of antioxidant components.


Assuntos
Adenosina/metabolismo , Aminoácidos/efeitos da radiação , Antioxidantes/efeitos da radiação , Cordyceps/efeitos da radiação , Ergocalciferóis/efeitos da radiação , Polissacarídeos/efeitos da radiação , Adenosina/efeitos da radiação , Aminoácidos/metabolismo , Antioxidantes/metabolismo , Cordyceps/química , Ergocalciferóis/metabolismo , Carpóforos/química , Carpóforos/efeitos da radiação , Micélio/química , Micélio/efeitos da radiação , Polissacarídeos/metabolismo
14.
Biochem Biophys Res Commun ; 486(2): 336-341, 2017 04 29.
Artigo em Inglês | MEDLINE | ID: mdl-28302483

RESUMO

Our previous studies revealed that CYP105A1 can convert vitamin D3 (VD3) to its active form, 1α,25-dihydroxyvitamin D3 (1,25D3). Site-directed mutagenesis of CYP105A1 based on its crystal structure dramatically enhanced its activity; the activity of double variants R73A/R84A and R73A/R84V was more than 100-fold higher than that of the wild type of CYP105A1. In contrast, these variants had a low ability to convert vitamin D2 (VD2) to 1α,25-dihydroxyvitamin D2 (1,25D2), whereas they catalyzed the sequential hydroxylation at positions C25 and C26 to produce 25,26D2. A comparison of the docking models of 25D2 and 25D3 into the substrate-binding pocket of R73A/R84A suggests that the side chain of the Met239 inhibits the binding of 25D2 for 1α-hydroxylation. Therefore, the Met239 residue of R73A/R84A was substituted for Ala. As expected, the triple variant R73A/R84A/M239A showed a 22-fold higher 1α-hydroxylation activity towards 25D2. To the best of our knowledge, this is the first report on the generation of microbial cytochrome P450 that converts VD2 to 1,25D2 via 25D2.


Assuntos
Proteínas de Bactérias/química , Sistema Enzimático do Citocromo P-450/química , Ergocalciferóis/química , Engenharia de Proteínas , Substituição de Aminoácidos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Clonagem Molecular , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Ergocalciferóis/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Hidroxilação , Simulação de Acoplamento Molecular , Mutagênese Sítio-Dirigida , Domínios Proteicos , Estrutura Secundária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Streptomyces/química , Streptomyces/enzimologia , Especificidade por Substrato
15.
Domest Anim Endocrinol ; 59: 44-52, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-27940098

RESUMO

In cattle, there are 2 significant forms of vitamin D: ergocalciferol (ERG) from fungi on roughage and cholecalciferol (CHO) from vitamin supplements or endogenous synthesis in the skin. The hypothesis of the present study is that vitamin D from the 3 sources is transported in different plasma fractions in the body. This is hypothesized to explain the lower efficiency of ERG compared to CHO in securing a sufficient plasma status of 25-hydroxyvitamin D and explain the inefficient excretion of dietary CHO into milk compared to endogenous CHO. Twenty vitamin D-depleted cows were assigned to 5 treatments: D2, housed indoor and fed 625-µg/d (25.000 IU) ERG; D3, housed indoor and fed 625-µg/d CHO; D2+D3, housed indoor and fed 625-µg/d ERG and 625-µg/d CHO; SUN, let out for daily pasture to facilitate CHO synthesis from sunlight; and D2+SUN, fed 625-µg/d ERG and let out for daily pasture. Blood samples were taken twice weekly and plasma fractionated by ultracentrifugation into 3 fractions: light lipoprotein (LLP), heavy lipoprotein (HLP), and protein and analyzed for content of ERG and CHO and their liver derived metabolites 25-hydroxyergocalciferol (25ERG) and 25-hydroxycholecalciferol (25CHO), respectively. Liver biopsies were taken on the last day of the study to asses gene expression related to vitamin D metabolism. During 4 wk of study, the vitamin D status in plasma increased to 19.3 to 22.8 ng/mL 25ERG in ERG-treated cows with the highest concentration in D2 (P ≤ 0.05) and to 25.0 to 33.4 ng/mL 25CHO in pasture or CHO-treated cows with the highest concentration in SUN (P ≤ 0.01). In plasma fractions, CHO was mainly found in the HLP fraction, whereas 25CHO was almost exclusively found in the protein fraction, probably due to its reported high binding affinity to vitamin D-binding protein. About 70% to 90% of 25ERG was found in the protein fraction and the remaining 25ERG was found in HLP, whereas ERG was found in both HLP and LLP fractions. In liver tissue, the expression of vitamin D-25-hydroxylase was lower in D2+D3 (P ≤ 0.05) and SUN (P ≤ 0.05) than that in the remaining groups, and the vitamin D receptor was expressed in the liver to a larger extent in D2+SUN than that in D2+D3 (P ≤ 0.05) and SUN (P ≤ 0.05). In conclusion, different plasma transport mechanisms may explain the lower physiological efficiency of ERG compared to CHO in securing the vitamin D status in plasma but do not explain the lower efficiency of synthetic CHO compared to endogenous CHO from sunlight or UV light in securing a high CHO content in milk.


Assuntos
Bovinos/sangue , Colecalciferol/sangue , Colecalciferol/metabolismo , Ergocalciferóis/sangue , Ergocalciferóis/metabolismo , Luz Solar , Animais , Colecalciferol/biossíntese , Feminino , Abrigo para Animais , Leite/química
16.
J Steroid Biochem Mol Biol ; 173: 168-172, 2017 10.
Artigo em Inglês | MEDLINE | ID: mdl-27825991

RESUMO

The putative biological activity of 24,25-(OH)2D remains unclear. Previous studies showed an increase in the circulating levels of this metabolite following fracture in chicks. Our laboratory has generated a mouse model deficient for the Cyp24a1 gene for studying the role of 24,25-(OH)2D. We set out to study the role of CYP24A1 and 24,25-(OH)2D in intramembranous bone formation during distraction osteogenesis in wild-type and Cyp24a1-deficient mice. Distraction osteogenesis was applied to mouse tibiae using a miniature external fixator apparatus. Histomorphometric parameters and gene expression differences between the mutant mice and the wild-type controls were measured using micro computed tomography and reverse-transcription quantitative PCR.There were no statistically significant differences between genotypes when bone volume/tissue volume ratios were calculated at mid distraction, end of distraction, mid consolidation, or end of consolidation. We measured reduced expression of the Col10a1 gene in the mutant vs. wild-type mice at mid distraction (0.4±0.1 vs. 1.0±0.2 respectively, p=0.01). Similarly, we measured a significantly lower expression of the osteogenic marker Atf4 in mutant vs. wild-type mice at end of distraction (0.7±0.1 vs. 1.0±0.1 respectively, p=0.01) and at mid consolidation (0.6±0.1 vs. 1.0±0.1 respectively, p=0.0003). These results suggest moderate and restricted differences in chondrogenesis and osteogenesis that did not affect the volume of bone produced following distraction. We conclude that CYP24A1 activity is not essential for intramembranous bone formation.


Assuntos
Regeneração Óssea , Deleção de Genes , Osteogênese , Vitamina D3 24-Hidroxilase/genética , Vitamina D3 24-Hidroxilase/metabolismo , Animais , Condrogênese , Ergocalciferóis/genética , Ergocalciferóis/metabolismo , Feminino , Genótipo , Masculino , Camundongos , Osteogênese por Distração
17.
Biochem Biophys Res Commun ; 473(4): 853-858, 2016 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-27037023

RESUMO

Our previous studies revealed that the double variants of CYP105A1- R73A/R84A and R73V/R84A-show high levels of activity with respect to conversion of vitamin D3 to its biologically active form, 1α,25-dihydroxyvitamin D3 (1α,25(OH)2D3). In this study, we found that both the double variants were also capable of converting vitamin D2 to its active form, that is, 1α,25-dihydroxyvitamin D2 (1α,25(OH)2D2), via 25(OH)D2, whereas its 1α-hydroxylation activity toward 25(OH)D2 was much lower than that toward 25(OH)D3. Comparison of the wild type and the double variants revealed that the amino acid substitutions remarkably enhanced both 25- and 26-hydroxylation activity toward vitamin D2. After 25-hydroxylation of vitamin D2, further hydroxylation at C26 may occur frequently without the release of 25(OH)D2 from the substrate-binding pocket. Thus, the double variants of CYP105A1 are quite useful to produce 25,26(OH)2D2 that is one of the metabolites of vitamin D2 detected in human serum.


Assuntos
Proteínas de Bactérias/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Ergocalciferóis/química , Ergocalciferóis/metabolismo , Engenharia de Proteínas , Proteínas de Bactérias/genética , Sistema Enzimático do Citocromo P-450/genética , Ativação Enzimática , Hidroxilação/fisiologia , Oxigenases de Função Mista/química , Oxigenases de Função Mista/metabolismo
19.
Alcohol Alcohol ; 51(1): 1-10, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26589585

RESUMO

AIMS: Heavy alcohol intake depletes the plasma vitamins due to hepatotoxicity and decreased intestinal absorption. However, moderate alcohol intake is often thought to be healthy. Therefore, effects of chronic moderate alcohol intake on liver and intestine were studied using urinary vitamin levels. Furthermore, effects of Tinospora cordifolia water extract (TCE) (hepatoprotective) on vitamin excretion and intestinal absorption were also studied. METHODS: In the study, asymptomatic moderate alcoholics (n = 12) without chronic liver disease and healthy volunteers (n = 14) of mean age 39 ± 2.2 (mean ± SD) were selected and divided into three groups. TCE treatment was performed for 14 days. The blood and urine samples were collected on Day 0 and 14 after treatment with TCE and analyzed. RESULTS: In alcoholics samples, a significant increase in the levels of gamma-glutamyl transferase, aspartate transaminase, alanine transaminase, Triglyceride, Cholesterol, HDL and LDL (P < 0.05) was observed but their level get downregulated after TCE intervention. Multivariate analysis of metabolites without missing values showed an increased excretion of 7-dehydrocholesterol, orotic acid, pyridoxine, lipoamide and niacin and TCE intervention depleted their levels (P < 0.05). In contrast, excretion of biotin, xanthine, vitamin D2 and 2-O-p-coumaroyltartronic acid (CA, an internal marker of intestinal absorption) were observed to be decreased in alcoholic samples; however, TCE intervention restored the CA and biotin levels. Vitamin metabolism biomarkers, i.e. homocysteine and xanthurenic acid, were also normalized after TCE intervention. CONCLUSION: Overall data depict that moderate alcohol intake is also hepatotoxic and decreases intestinal absorption. However, TCE treatment effectively increased the intestinal absorption and retaining power of liver that regulated alcohol-induced multivitamin deficiency.


Assuntos
Alcoolismo/metabolismo , Trato Gastrointestinal/efeitos dos fármacos , Absorção Intestinal/efeitos dos fármacos , Fígado/efeitos dos fármacos , Extratos Vegetais/farmacologia , Tinospora , Vitaminas/metabolismo , Adulto , Biotina/metabolismo , Estudos de Casos e Controles , Ergocalciferóis/metabolismo , Trato Gastrointestinal/metabolismo , Homocisteína/metabolismo , Humanos , Fígado/metabolismo , Índice de Gravidade de Doença , Tartronatos/metabolismo , Vitaminas/sangue , Vitaminas/urina , Xantina/metabolismo , Xanturenatos/metabolismo
20.
J Dairy Res ; 83(1): 67-71, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26608575

RESUMO

Vitamin D has become one of the most discussed nutrients in human nutrition, which has led to an increased interest in milk as a vitamin D source. Problems related to fortifying milk with synthetic vitamin D can be avoided by securing a high content of natural vitamin D in the milk by supplying dairy cows with sufficient vitamin D. However, choosing the most efficient route and form of supplementation requires insight into how different vitamin D metabolites are transported in the body of cattle. There are two forms of vitamin D: vitamin D2 (D2) and vitamin D3 (D3). Vitamin D2 originates from fungi on roughage. Vitamin D3 originates either from endogenous synthesis in the skin or from feed supplements. Vitamin D2 is chemically different from, and less physiologically active than, D3. Endogenous and dietary D3 is chemically similar but dietary D3 is toxic, whereas endogenous D3 appears well regulated in the body.


Assuntos
Ração Animal/análise , Bovinos/sangue , Colecalciferol/metabolismo , Ergocalciferóis/metabolismo , Luz Solar , Vitamina D/análogos & derivados , Fenômenos Fisiológicos da Nutrição Animal , Animais , Colecalciferol/química , Dieta/veterinária , Suplementos Nutricionais , Ergocalciferóis/química , Masculino , Vitamina D/sangue , Vitaminas/química , Vitaminas/metabolismo
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