A deficiência de vitamina D (dVD) aumenta o risco de morte em pacientes hospitalizados. A injúria de isquemia/reperfusão renal (Isq) ativa vias de necrose e/ou apoptose e proliferação celular. A injúria renal aguda (IRA) induz a ativação de inibidores do ciclo celular, incluindo a p21, uma inibidora de kinase dependente de ciclina, a qual possui efeito protetor na IRA. A p21 é um alvo genômico da 25-hidroxivitamina D [25 (OH) D], a qual, atuando através de receptores de vitamina D (VDRs), possui efeitos imunomoduladores potentes e antiproliferativos, sugerindo a participação deste hormônio na fisiopatologia da doença renal. Desta forma, o objetivo deste estudo foi verificar a participação da deficiência de vitamina D no modelo de isquemia/reperfusão renal em ratos. Foram utilizados ratos Wistar que foram divididos em quatro grupos: controle (C), animais que receberam dieta padrão por 30 dias; dVD, animais que receberam dieta livre de vitamina D por 30 dias; Isq, animais que receberam dieta padrão por 30 dias e no 28º dia foram submetidos ao insulto de isquemia/reperfusão em ambos os rins por 45 minutos; e dVD+Isq, animais que receberam dieta livre de vitamina D por 30 dias e no 28º dia foram submetidos ao insulto de isquemia/reperfusão em ambos os rins por 45 minutos. Ao final dos 30 dias e após 48 horas da realização da isquemia/reperfusão, os animais foram submetidos à eutanásia e amostras de sangue, urina e tecido renal foram coletados para o estudo dos mecanismos de lesão renal. A injúria renal aguda associada à deficiência de vitamina D levou a uma queda da filtração glomerular e aumento da proteinúria; aumento da relação peso renal/peso corporal, sugerindo maior proliferação e hipertrofia; induziu uma diminuição na ativação dos receptores de vitamina D e da expressão da proteína p21 e aumento da expressão de caspase-3; observou-se déficit de concentração urinária com diminuição da expressão da AQP2; e um maior dano morfológico caracterizado pela análise da área intersticial e presença de necrose tubular. Nossos dados mostraram que alterando os níveis da p21 na IRA isquêmica, a vitamina D, via VDRs, controla a inflamação renal, a proliferação e a lesão celular / Vitamin D deficiency (VDD) increases the risk of death in hospitalized patients. Ischemia/reperfusion injury activates pathways of necrosis and/or apoptosis and cell proliferation. Acute kidney injury (AKI) induces the activation of cell cycle inhibitors, including p21, an inhibitor of cyclin-dependent kinase, which has a protective effect on the IRA. The p21 is a genomic target of 25-hydroxyvitamin D [ 25 (OH) D], which, acting through vitamin D receptors (VDRs), has potent antiproliferative and immunomodulatory effects, suggesting the involvement of this hormone in the pathophysiology of renal disease. Thus, the aim of this study was to assess the role of vitamin D deficiency in rats submitted to renal ischemia/reperfusion. Wistar rats were divided into four groups: control (C), animals that received a standard diet for 30 days; VDD, animals that received vitamin D-free diet for 30 days; IRI, animals that received standard diet for 30 days and on day 28 were subjected to the ischemia/reperfusion insult (IRI) in both kidneys for 45 minutes; and VDD+IRI, animals that received vitamin D-free diet for 30 days and on day 28 were subjected to the IRI in both kidneys for 45 minutes. At the end of 30 days and 48 hours after the IRI insult, the animals were euthanized and samples of blood, urine and kidney tissue were collected to study the mechanisms of renal injury. Acute kidney injury associated with vitamin D deficiency led to a decrease in glomerular filtration rate and increased proteinuria; increased relative kidney weight/body weight, suggesting greater proliferation and hypertrophy; induced a decrease in the activation of vitamin D receptors and the p21 protein expression and, increased caspase-3 expression; renal concentration impairment with decreased AQP2 expression, as well as greater morphological damage characterized by interstitial area analysis and presence of tubular necrosis. Our data showed that altering the levels of p21 in ischemic-AKI, vitamin D via VDRs, controls kidney inflammation, proliferation and cell injury

No description available.

A deficiência de vitamina D (dVD) aumenta o risco de morte em pacientes hospitalizados. A injúria de isquemia/reperfusão renal (Isq) ativa vias de necrose e/ou apoptose e proliferação celular. A injúria renal aguda (IRA) induz a ativação de inibidores do ciclo celular, incluindo a p21, uma inibidora de kinase dependente de ciclina, a qual possui efeito protetor na IRA. A p21 é um alvo genômico da 25-hidroxivitamina D [25 (OH) D], a qual, atuando através de receptores de vitamina D (VDRs), possui efeitos imunomoduladores potentes e antiproliferativos, sugerindo a participação deste hormônio na fisiopatologia da doença renal. Desta forma, o objetivo deste estudo foi verificar a participação da deficiência de vitamina D no modelo de isquemia/reperfusão renal em ratos. Foram utilizados ratos Wistar que foram divididos em quatro grupos: controle (C), animais que receberam dieta padrão por 30 dias; dVD, animais que receberam dieta livre de vitamina D por 30 dias; Isq, animais que receberam dieta padrão por 30 dias e no 28º dia foram submetidos ao insulto de isquemia/reperfusão em ambos os rins por 45 minutos; e dVD+Isq, animais que receberam dieta livre de vitamina D por 30 dias e no 28º dia foram submetidos ao insulto de isquemia/reperfusão em ambos os rins por 45 minutos. Ao final dos 30 dias e após 48 horas da realização da isquemia/reperfusão, os animais foram submetidos à eutanásia e amostras de sangue, urina e tecido renal foram coletados para o estudo dos mecanismos de lesão renal. A injúria renal aguda associada à deficiência de vitamina D levou a uma queda da filtração glomerular e aumento da proteinúria; aumento da relação peso renal/peso corporal, sugerindo maior proliferação e hipertrofia; induziu uma diminuição na ativação dos receptores de vitamina D e da expressão da proteína p21 e aumento da expressão de caspase-3; observou-se déficit de concentração urinária com diminuição da expressão da AQP2; e um maior dano morfológico caracterizado pela análise da área intersticial e presença de necrose tubular. Nossos dados mostraram que alterando os níveis da p21 na IRA isquêmica, a vitamina D, via VDRs, controla a inflamação renal, a proliferação e a lesão celular / Vitamin D deficiency (VDD) increases the risk of death in hospitalized patients. Ischemia/reperfusion injury activates pathways of necrosis and/or apoptosis and cell proliferation. Acute kidney injury (AKI) induces the activation of cell cycle inhibitors, including p21, an inhibitor of cyclin-dependent kinase, which has a protective effect on the IRA. The p21 is a genomic target of 25-hydroxyvitamin D [ 25 (OH) D], which, acting through vitamin D receptors (VDRs), has potent antiproliferative and immunomodulatory effects, suggesting the involvement of this hormone in the pathophysiology of renal disease. Thus, the aim of this study was to assess the role of vitamin D deficiency in rats submitted to renal ischemia/reperfusion. Wistar rats were divided into four groups: control (C), animals that received a standard diet for 30 days; VDD, animals that received vitamin D-free diet for 30 days; IRI, animals that received standard diet for 30 days and on day 28 were subjected to the ischemia/reperfusion insult (IRI) in both kidneys for 45 minutes; and VDD+IRI, animals that received vitamin D-free diet for 30 days and on day 28 were subjected to the IRI in both kidneys for 45 minutes. At the end of 30 days and 48 hours after the IRI insult, the animals were euthanized and samples of blood, urine and kidney tissue were collected to study the mechanisms of renal injury. Acute kidney injury associated with vitamin D deficiency led to a decrease in glomerular filtration rate and increased proteinuria; increased relative kidney weight/body weight, suggesting greater proliferation and hypertrophy; induced a decrease in the activation of vitamin D receptors and the p21 protein expression and, increased caspase-3 expression; renal concentration impairment with decreased AQP2 expression, as well as greater morphological damage characterized by interstitial area analysis and presence of tubular necrosis. Our data showed that altering the levels of p21 in ischemic-AKI, vitamin D via VDRs, controls kidney inflammation, proliferation and cell injury

A vitamina D (VD) pode estar envolvida no controle da proliferação, diferenciação e apoptose em linhagens mamárias. Existem evidências de que mulheres com câncer de mama apresentam menor concentração sérica de 25(OH)D3 ou de 1,25(OH)2D3 em relação às mulheres sem câncer. Por outro lado, pouco se sabe se a concentração sérica de VD pode influenciar o desenvolvimento de câncer de mama em cadelas e se o hormônio pode ter efeito quimiopreventivo, inibindo o aparecimento de tumores ou mesmo efeito terapêutico, reduzindo a proliferação de células malignas. Logo, nossos objetivos foram comparar a concentração sérica de 25(OH)D3 em animais com e sem tumor mamário e analisar as ações de 1,25(OH)2D3 em glândulas mamárias normais e tumorais de cadelas, utilizando como modelo a cultura de tecidos. Inicialmente foram incluídas 39 cadelas portadoras de tumor de mama e 64 cães sem tumor (controle), sendo que 50 eram fêmeas e 14 eram machos. Observamos que os animais do grupo tumoral possuíam idade mais avançada (mediana de 108 meses) em relação ao grupo controle (mediana de 36 meses para os machos e 24 meses para as fêmeas). No grupo controle, a concentração sérica foi maior nos machos (32,5 ± 19,3 ng/mL) do que nas fêmeas (22,8 ± 9,6 ng/mL), mas não houve diferença em relação ao grupo tumor (26,62 ± 14,25 ng/mL). Em relação à dieta, a concentração sérica de 25(OH)D3 foi maior nas fêmeas do grupo controle que se alimentavam de ração em comparação às que se alimentavam de comida caseira e ração. Entretanto, não houve diferença em relação à exposição ao sol e características da pelagem em todos os animais. No grupo tumoral, houve correlação inversa da concentração sérica de 25(OH)D3 em relação à idade, mas não houve diferença quanto ao tipo histológico ou estadiamento da doença. Foram coletadas 70 amostras de tumor de mama e de tecido mamários normal de cadelas, as quais foram cultivadas em fatias. Dos tecidos tumorais, 50% eram positivos para receptor de estrógeno (acima de 10% de células marcadas) e 44% eram positivos para HER-2 (método HercepTest). Detectou-se expressão gênica e proteica do receptor da vitamina D (VDR) em tecido mamário normal e tumoral, sendo identificado três padrões na imunoistoquímica: I - células epiteliais e mioepiteliais (mais frequentemente encontrada em tecido normal), II - marcação predominante em células mioepiteliais (mais comum em tecido tumoral e III - marcação predominante em células epiteliais. As amostras foram tratadas com 1,25(OH)2D3 nas concentrações 0,228 nM, 2 nM e 100 nM (concentração fisiológica, farmacológica que não induz hipercalcemia e farmacológica que induz hipercalcemia, respectivamente). O conteúdo de VD tecidual avaliado por cromatografia líquida foi crescente de acordo com as concentrações de VD utilizadas, indicando penetração do hormônio nas fatias. Observou-se indução da expressão de CYP24A1, que variou de 27 a 158 vezes dependendo da concentração utilizada, indicando ativação genômica da via da VD e que o tecido permanece metabolicamente ativo em cultura. Entretanto, não houve diferença da expressão gênica de outros genes envolvidos com o metabolismo da VD (CYP27B1), genes envolvidos no controle da proliferação (CDKN1A e CDKN1B) e genes envolvidos com a imunidade (CD14). O tratamento com calcitriol nas diferentes concentrações não induziu a apoptose (expressão proteica de caspase-3 clivada) e não alterou a proliferação nos tecidos normais (expressão proteica de Ki-67), mas diminuiu a proliferação nos tecidos tumorais. Não foi observada correlação entre a indução da apoptose e redução da proliferação com os padrões de expressão proteica de VDR. Concluindo, não observamos diferença na concentração sérica de 25(OH)D3 entre cadelas com tumor de mama e animais controle. Detectamos que o calcitriol em concentração fisiológica ativa a via genômica de VD em mama normal e tumoral e reduz o índice de proliferação (expressão de Ki-67) nos tumores de mama / Vitamin D (VD) may be involved in the control of proliferation, differentiation and apoptosis of mammary cell lines exposed to high concentrations of the hormone. There is some evidence that women with breast cancer present lower serum level of 25(OH)D3 or 1,25(OH)2D3 compared to women without cancer. Moreover, little is known if serum concentration of VD can influence the development of mammary tumors in dogs and if the hormone may have chemopreventive effect by inhibiting the appearance of tumors or therapeutic effect, reducing the proliferation of malignant cells. Therefore, our goals were to compare the serum 25(OH)D3 level in animals with and without mammary tumor and to analyze 1,25(OH)2D3 effects in normal and tumoral canine mammary glands, using as a model the tissue culture. At first, 39 bitches with mammary tumor and 64 dogs without tumor (control), of which 50 were females and 14 were males were included. Animals in tumor group were older (median 108 months) compared to control group (median 36 months for males and 24 months for females). In control group, serum concentration was higher in males (32.5 ± 19.3 ng/mL) than in females (22.8 ± 9.6 ng/mL), but there was no difference when compared to tumor group (26.62 ± 14.25 ng/mL). Regarding diet, serum 25(OH)D3 level was higher in control bitches fed commercial pet food compared to those fed homemade and commercial pet food combined. However, there was no difference of serum 25(OH)D3 concentration, sun exposure and coat features. In tumor group, there was an inverse correlation between serum 25(OH)D3 and age, but there was no difference in 25(OH)D3 concentration among bitches with different histological type or clinical stage of the disease. Seventhy bitches diagnosed with mammary tumors had tumor and mammary samples collected, sliced and cultured. In tumor tissues, 50% were positive for estrogen receptor (over 10% of cells stained), and 44% were positive for HER-2 (HercepTest method). Vitamin D receptor (VDR) protein and genic expression was detected in normal and tumoral samples. Three patterns of VDR were detected by immunohistochemistry: I - localizated in epithelial and myoepithelial cells (more often in normal tissues), II - predominantly in myoepithelial cells (most common in tumor tissues) and III - predominantly in epithelial cells. Normal anmammary slices were treated with 1,25(OH)2D3 0.228 and 100nM concentrations (concentration physiological and pharmacological, respectively) and mammay tumor slices were treated with 1,25(OH)2D3 2 nM concentrations (drug concentration which does not induce hypercalcemia) and 100 nM, for 24 hours. VD tissue content measured by liquid chromatography was higher in samples exposed to high VD concentration, indicating penetration of the hormone in slices. VD treatment induced CYP24A1 expression, 27 to 158 fold depending on the concentration used, and indicating activation of the VD genomic pathway. This result also suggest that the tissue remains metabolically active in culture. However, no difference in gene expression of other target genes such as CYP27B1, genes involved in proliferation as CDKN1A and CDKN1B and genes involved in immunity, such as CD14. Calcitriol treatment at different concentrations did not induce apoptosis (protein expression of cleaved caspase-3) and did not alter proliferation in normal tissues (expression of protein Ki -67), but decreased proliferation in tumor tissues. No correlation was observed between the induction of apoptosis and reduction of proliferation with the protein expression patterns of VDR. In conclusion, no difference in serum 25(OH)D3 between bitches with mammary tumor and control animals was observed. In normal and tumoral mammary samples calcitriol physiologic concentration activated VD genomic pathway and in tumor samples calcitriol reduced the proliferation index (Ki-67)

No description available.

Vitamin-D-Mangel und -Unterversorgung haben sich zu einem großen Problem für die öffentliche Gesundheit entwickelt, das vor allem auf eine unzureichende Sonnenlichtexposition und eine begrenzte Zufuhr dieses essenziellen Nährstoffs über die Nahrung zurückzuführen ist. Diese "Pandemie" hat zu einem bemerkenswerten Anstieg der Nachfrage nach der Bestimmung des zirkulierenden 25-Hydroxyvitamin-D-Spiegels (25(OH)D) geführt. Die meisten Studien fokussieren sich in erster Linie auf 25(OH)D und betrachten es als Biomarker für den Vitamin-D-Status. Es besteht jedoch ein wachsendes Interesse an der gleichzeitigen Messung mehrerer klinisch bedeutsamer Vitamin-D-Metaboliten wie dem nativen Vitamin D, 25(OH)D-Epimeren, 1,25-Dihydroxyvitamin D und 24,25-Dihydroxyvitamin D. Die Flüssigchromatographie/Tandem-Massenspektrometrie (LC-MS/MS) gilt als "Goldstandard" für die Bestimmung des Vitamin-D-Spiegels und ermöglicht die gleichzeitige Analyse mehrerer Analyten, die für das Verständnis der physiologischen Rolle von Vitamin D und seiner klinischen Auswirkungen unerlässlich sind. In der vorliegenden Doktorarbeit soll verschiedene Aspekte der Vitamin-D-Landschaft erforschen, umfassende Einblicke geben und Herausforderungen innerhalb des Forschungsthemas besprechen. Zunächst lag der Schwerpunkt auf dem Vergleich mehrerer Derivatisierungsreagenzien für die Vitamin-D-Analyse und der Frage, wie sie sich auf die Nachweisempfindlichkeit der Methode und die chromatographische Trennung der getesteten Vitamin-D-Metaboliten auswirken. Ein zweiter Schwerpunkt lag auf der Untersuchung der Stabilität der Derivatisierungsprodukte in Serumextrakten. Der letzte Versuch war die Entwicklung einer LC-MS/MS-Methode, die die Ergebnisse der vorangegangenen Untersuchungen nutzen sollte. Als Ergebnis wurde eine neue Methode vorgestellt, die meines Wissens zum ersten Mal FMP-TS für die chemische Derivatisierung von Vitamin-D-Metaboliten verwendet. / Vitamin D deficiency has emerged as a significant public health concern, attributed largely to insufficient exposure to sunlight and limited dietary sources rich in this essential nutrient. This “pandemic” has led to a notable increase in the demand for assessing circulating levels of 25-hydroxyvitamin D (25(OH)D). Most studies have primarily focused on 25(OH)D, considered it as the vitamin D status biomarker. However, there is a growing interest in simultaneously measuring multiple clinically significant vitamin D metabolites such as the native vitamin D, 25(OH)D epimer, 1,25-dihydroxyvitamin D, 24,25-dihydroxyvitamin D and others. Thus, there is an urgent need to develop analytical methods which will be able to separate and quantify the metabolites of interest accurately and precisely. Liquid chromatography/tandem mass spectrometry (LC-MS/MS) stands out as the “gold standard” technique for vitamin D analysis. Its versatility enables simultaneous analysis of multiple analytes, thereby facilitating the generation of comprehensive vitamin D profiles essential for understanding its physiological role and clinical implications. The present doctoral thesis is designed to explore diverse aspects of vitamin D landscape, providing comprehensive insights and addressing challenges within the research topic. Initially there was a focus on comparing multiple derivatization reagents for vitamin D analysis and discovering how they impact the method’s detection sensitivity and the chromatographic separation of the tested vitamin D metabolites. Moreover, there was a second focal point on examining the stability of the derivatization products in serum extracts. The final attempt was the development of an LC-MS/MS method that would utilize the outcomes of the investigations conducted before. As a result, a new method was introduced utilizing FMP-TS for the chemical derivatization of vitamin D metabolites, a reagent which has not been reported before for vitamin D analysis.

Die Erkrankungen des atopischen Formenkreises sind durch eine hypererge Immunreaktion auf harmlose Umwelt-Antigene gekennzeichnet und umfassen u.a. die klinischen Bilder der allergischen Rhinokonjunktivitis, des allergischen Asthmas und der atopischen Dermatitis. Die Pathogenese atopischer Krankheiten ist zwar noch nicht hinreichend aufgeklärt, aber man schreibt der Regulation des IgE bei der Entstehung und Erhaltung dieser Krankheiten eine zentrale Rolle zu. Auf der Suche nach potentiellen anti-allergischen Therapeutika wurde in dieser Arbeit erstmalig der Einfluß von 1alpha,25-Dihydroxyvitamin-D3 (VD) und dem niedrigkalzämischen Derivat EB1089 in vitro auf die IgE-Produktion humaner B-Zellen gesunder Spender untersucht, die zuvor mit anti-CD40+IL-4 stimuliert wurden, wodurch der Isotypenklassenwechsel nativer B-Zellen nach IgE resultierte. Das Steroidhormon VD ist für die Kalziumhämostase essentiell, jedoch sind die immunregulatorischen Funktionen des VD noch relativ unerforscht. Die Experimente dieser Arbeit zeigen eine starke, dosisabhängige Hemmung der IgE-Produktion in diesem Modell, die spezifisch von VD und dem Derivat EB1089 vermittelt wird. Auch bei PBMC atopischer Spender konnte die basale IgE-Produktion mit VD dosisabhängig signifikant gehemmt werden. Eine mögliche Proliferationshemmung, die diese Wirkung erklären könnte, konnte ausgeschlossen werden. Die Messung der Produktion der anderen Immunglobulinklassen (Ig) IgA, IgG und IgM bestätigte, daß die Zellviabilität und die Ig-Produktion außer der von IgE durch VD und EB1089 in diesem Modell nur wenig beeinflußt wird. Diese Beobachtungen weisen auf eine selektive, spezifische Inhibition der IgE-Synthese durch das Molekül aus der Familie der Steroidhormone hin. Auf der Suche nach möglichen indirekten Mechanismen der IgE-Regulation, die die VD-vermittelte Inhibition der IgE-Synthese erklären könnten, wurde der Einfluß von VD auf die Expression der kostimulatorischen Faktoren der IgE-Produktion CD23 (Fc(-RII), CD54 (ICAM-1) und CD86 (B7.2) sowie auf die Sekretion der Zytokine, IL-6, sCD23 und IFN-gamma untersucht. Die Ergebnisse dieser Arbeit deuteten jedoch nicht auf eine Beteiligung der genannten Faktoren an der Regulation der VD-vermittelten Hemmung der IgE-Produktion hin. Diese in vitro Arbeit stellt einen wichtigen Hinweis auf eine potentielle Nutzung von VD in der anti-allergischen Therapie dar, jedoch deutet die beobachtete Hemmung der IgE-Synthese von PBMC atopischer Spender darauf hin, daß auch in vivo die IgE-Synthese gehemmt werden kann. Es sind daher weitere Studien des Signalweges des VD auf molekularer Ebene, sowie in vivo Versuche notwendig, um zu klären, ob VD selbst oder ein möglichst nicht-kalzitropes VD-Derivat mit ausgeprägten immunologischen Eigenschaften für die anti-allergische Therapie geeignet ist. / Atopic diseases are characterized by a hyperergic immunoreaction towards harmless environmental antigens, expressed by the clinical phenotype of allergic rhinitis, allergic asthma and atopic dermatitis. Even though the pathogenesis of atopic diseases are not completely clear, the regulation of IgE plays a crucial role in the development and maintenance of these disorders. In search of new potentially anti-allergic drugs, the effect of 1alpha,25-Dihydroxyvitamin-D3 (VD) and the low-calcemic synthetic analogue EB1089 was determined in anti-CD40+Il-4-mediated IgE-production by B-cells of healthy donors in vitro. The essential role of VD in calcium hemostasis is well understood, but the immunoregulatory functions of the hormone are still quite unclear. The present data show for the first time, that VD and the analogue EB1089 mediate a strong, dose-dependent inhibition of IgE-synthesis. In PBMC of atopic donors, the baseline IgE-production was also significantly inhibited by VD. Anti-CD40+IL-4-mediated proliferation of B-cells in the presence of VD was only modestly modulated, as well as the production of the other immunoglobulin-isotypes IgA, IgG and IgM. Taken together these data suggest a specific and selective inhibition of IgE-synthesis by VD and EB1089. In order to explore possible mechanisms of VD induced IgE-inibition, the expression of CD23 (Fc(-RII), CD54 (ICAM-1) and CD86 (B7.2) on anti-CD+IL-4 stimulated B-cells was determined as well as IL-6, sCD23 und IFN-gamma. However, none of these molecules were significantly modulated in the presence of VD. Taken together, these in vitro experiments suggest a potential role of VD as a new anti-allergic drug, which is underlined by the observed inhibition of IgE-production by PBMC in allergic donors.

Background A well-balanced nutritious diet is important for the pregnant woman and the growing fetus, as well as for their future health. Poor nutrition results from both over-consumption of energy-rich foods which can lead to a higher weight gain than is healthy and under-nutrition of essential nutrients. Food intake is regulated in complex biological systems by many factors, where steroid hormone is one factor involved. The overall aim of this thesis is to describe dietary intake, vitamin D levels, dietary information and dietary changes, and to study the relation between allopregnanolone and weight gain during pregnancy and postpartum. Methods Study I was a qualitative study with focus group interviews with 23 pregnant women. The text was analysed with content analysis. Study II was a quantitative cross-sectional study conducted in early pregnancy (n=209) with a reference group (n=206). Self-reported dietary data from a questionnaire was analysed using descriptive comparative statistics and a cluster analysis model (Partial Least Squares modelling). Study III had a quantitative longitudinal design. Vitamin D concentrations were analysed in 184 women, collected on five occasions during pregnancy and postpartum. Descriptive comparative statistics and a linear mixed model were used. Study IV was a quantitative longitudinal study with 60 women. Concentrations of allopregnanolone were analysed in gestational week 12 and 35. Descriptive and comparative statistics as well as Spearman’s correlation (rho) were used to describe the relationship between weight gain and allopregnanolone concentrations.   Results The focus group interviews showed that women wanted to know more about different foods to reduce any risk for their child but the information about foods was partly up to themselves to find out. They expressedfeelingsof insecurityand guiltif they accidentallyate something“forbidden”. The recommendationswere followedas best as possiblealong withcommon sense todeal with dietchanges. The main themes were “Finding out by oneself”, “Getting professional advice when health problems occur”, “Being uncertain” and “Being responsible with a pinch of salt”. Some differences in the dietary patterns were found among the pregnant women compared to references, with less, vegetables (47 g/day), potatoes/rice/pasta (31 g/day), meat/fish (24 g/day) and intake of alcohol and tobacco/snuff but a higher intake of supplements. Bothpregnant women and referenceshad intakes offolatethrough diet45% (pregnant) and 22% (references) lower than current recommendations(500vs400g/day). Vitamin Dintake was34% lower than the recommendationsof 10mg/day. At least a third of the participants had insufficient plasma levels below 50 nmol/L of vitamin D. Season was a strong factor influencing the longitudinal pattern. Gestational week, season, total energy intake, dietary intake of vitamin D, and multivitamin supplementation over the previous 14 days were factors related to vitamin D levels. A correlation betweenallopregnanoloneconcentrations ingestationalweek 35and weight gainin weeks12–35was seen (p = 0.016). Therewas alsoa correlation betweenthe increase inallopregnanolone(weeks12–35) andweight gain(see above) (p = 0.028).   Conclusions Dietary recommendations were described as contradictory and confusing and the dietary advice felt inadequate. The women faced their diet changes and sought information on their own but would have wished for more extensive advice from the midwife. The intake of vitamins essential for pregnancy was lower than recommended, which is also confirmed by low plasma levels of vitamin D in at least one third of the pregnant women. Vitamin D levels peaked in late pregnancy. Aside from gestational week and season which were related to plasma levels, intake from foods and supplements also affected the levels. Reasons for weight gain are complex and depend on many factors. Allopregnanolone is a factor that was seen to relate to the weight gain of the studied pregnant women. / Bakgrund En välbalanserad näringsrik kost är viktig för den gravida kvinnan och det växande fostret, så även för deras framtida hälsa. En bristfällig kost kan utgöras av både överförbrukning av energirika livsmedel vilket kan leda till högre viktuppgång än vad som är hälsosamt och bristande intag av viktiga näringsämnen. Kostintag regleras av komplexa biologiska system där flera faktorer är inblandade däribland steroidhormonet allopregnanolon. Det övergripande syftet med denna avhandling är att under och efter graviditet beskriva kostintag, vitamin D-nivåer, kostinformation och kostförändringar och att studera allopregnanolons relation till viktökning. Metod Studie I var en kvalitativ studie med fokusgruppsintervjuer med 23 gravida kvinnor. Texten analyserades med innehållsanalys. Studie II var en kvantitativ tvärsnittsstudie som genomfördes i tidig graviditet (n = 209) och med en grupp icke-gravida kvinnor (kontrollgrupp) (n=206). Självrapporterade kostdata från ett frågeformulär analyserades med beskrivande, jämförande statistik och en klusteranalysmodell (Partial Least Squares modellering). Studie III hade en kvantitativ longitudinell design. Vitamin D-koncentrationer analyserades hos 184 kvinnor, vid fem tillfällen under graviditeten och efter förlossningen. Beskrivande, jämförande statistik och en linjär mixad regressionsmodell användes. Studie IV var en kvantitativ longitudinell studie med 60 kvinnor. Koncentrationerna av allopregnanolon analyserades vid graviditetsvecka 12 och 35. Beskrivande och jämförande statistik samt Spearman’s korrelation användes för att beskriva samband mellan viktökning och koncentrationer av allopregnanolon. Resultat Intervjuerna i studie I visade att kvinnor ville veta mer om olika typer av mat för att minska en eventuell risk för sina barn men kostinformation var delvis upp till dem själva att ta reda på. De VIII uttryckte känslor av osäkerhet och skuld om de råkat äta något ”förbjudet”. Rekommendationerna följdes så väl som möjligt, tillsammans med sunt förnuft för att hantera kostförändringar. Huvudteman var ”Söka information på egen hand”, ”Få professionell rådgivning när problem uppstår”, ”Känna sig osäker” och ”Ta ansvar med en nypa salt”. I studie II kunde man se vissa skillnader i kostmönster bland de gravida kvinnorna jämfört med kontrollgruppen: mindre intag av grönsaker (47 g/dag), potatis/ris/pasta (31 g/dag), kött/fisk (24 g/dag) och alkohol och tobak/snus och ett högre intag av kosttillskott. Både gravida kvinnor och kontrollgruppen hade lägre intag av folsyra via kosten med 45 % (gravida) och 22 % (kontrollgruppen) än de gällande rekommendationer som är (500 resp 400 g/dag). I studie III såg man att inta et av vitamin D var 34 % lägre än rekommendationen på 10 µg/dag. Minst en tredjedel av deltagarna hade otillräckliga plasma nivåer av vitamin D, under 50 nmol/L. Årstid var en stark faktor som påverkar det longitudinella mönstret. Graviditetsvecka, säsong, totala energiintaget, intaget av vitamin D och multivitamintillskott under de senaste 14 dagarna var faktorer som relaterade till Dvitaminnivåer. I studie IV sågs ett samband mellan allopregnanolon-koncentrationer vid graviditetsvecka 35 och viktökning från vecka 12 till 35 (p = 0,016). Det sågs också ett samband mellan ökningen av allopregnanolon (vecka 12–35) och viktökningen (se ovan) (p = 0,028). Slutsatser Kostrekommendationer beskrevs som motsägelsefulla och förvirrande och kostråden de fick uppfattades som otillräckliga. Kvinnorna tog itu med sina kostförändringar och sökte information på egen hand men hade önskat mer omfattande råd från barnmorskan. Intaget av vitaminer viktiga för graviditeten var lägre än rekommendationerna, vilket också bekräftas av låga plasmanivåer av D-vitamin hos cirka en tredjedel av de gravida kvinnorna. D-vitaminnivåerna nådde en topp i slutet av graviditeten. Graviditetsvecka och säsong på året påverkade D vitaminnivåer, så även intag via mat och kosttillskott. Orsakertill viktökning är komplexa och beror på många faktorer. Allopregnanolon är en faktor som sågs relatera till viktökningen hos de undersökta gravida kvinnorna.

Background: Vitamin D is crucial for skeletal muscle function. 25-hidroxyvitamin D (25(OH)D) has been correlated with skeletal muscle mass and intramyocellular lipid (IMCL) content. The purpose of this study was to understand how calcitriol, the active vitamin D metabolite, directly affects myocellular size and lipid partitioning. Methods: C2C12 myotubes were treated with calcitriol (100nM) or vehicle control for 24 or 96 h. Myotube diameter and protein synthesis rate were measured to determine effects of calcitriol on myocellular size. Intramyocellular triacylglycerol (IMTG), diacylglycerol (DAG), and ceramide content were measured by LC/MS. Expression of genes involved in lipid packaging and lipolysis were measured by RT-PCR. Insulin-stimulated phosphorylated Akt (Thr 308) was determined by western blot. Results: Calcitriol did not affect myocellular size or protein synthesis rate. Calcitriol increased total DAG and ceramides in a sub-species specific manner. Calcitriol increased IMTG area, but did not affect total IMTG content. Calcitriol reduced mRNA content of diglyceride acyltransferase and increased mRNA content of lipid packaging genes. Calcitriol did not negatively affect insulin-stimulated pAkt. Conclusions: These results suggest calcitriol directly alters lipid content and packaging in skeletal muscle cells. Altering the expression of lipid packaging genes and increasing IMCL subspecies content may be mechanisms by which vitamin D improves skeletal muscle function in vivo.

<p>Human physiology is dependent on an accurate phosphate (Pi) homeostasis. Defective Pi regulation causes hyper- or hypophosphatemia, which are associated with ectopic calcification or impaired bone mineralization, and a shortened life span. Current endocrine models of Pi homeostasis are incomplete. However, studies of acquired and hereditary disorders of Pi homeostasis have revealed new potential Pi regulating hormones, Phosphatonin(s). One of these is fibroblast growth factor-23 (FGF23). FGF23 is produced in bone and is secreted into the circulation. Mutations in FGF23 causes disturbed Pi regulation, without the appropriate counter-regulatory actions of parathyroid hormone or vitamin D. By the generation of FGF23 transgenic mice, which display phenotypic similarities to patients with hypophosphatemic disorders, we show that FGF23 exerts endocrine actions in the kidney and causes osteomalacia. Renal FGF23 actions severely decrease Pi reabsorption and expression of Klotho, a suggested age suppressor gene, known to be crucial in FGF23 receptor binding and activation. In bone, our transgenic model displays impaired osteoclast polarization, which should be detrimental to osteoclastic bone resorption in osteomalacia. However, in our model osteoclasts efficiently participate in bone matrix degradation. Furthermore, we investigated a large population-based cohort in order to elucidate the role of FGF23 in normal physiology. Importantly, we were able to demonstrate an association of FGF23 to parathyroid hormone, renal function and bone mineral density and we found a correlation of FGF23 to weight and body fat mass. The studies on which this thesis is based, demonstrate that FGF23 has phosphatonin-like properties and that the skeleton functions as an endocrine organ. In addition, the results indicate that FGF23 has a role in bone mineral and lipid metabolism, and that FGF23 is a possible diagnostic marker and therapeutic target for the future.</p>