Die Expression und Regulation der CYP3A in menschlicher Haut

In Leber und Dünndarm bauen CYP3A-Enzyme eine Vielzahl von Fremdstoffen ab, die in den Körper gelangt sind. Zudem aber sind diese Enzyme auch in anderen Organen, wie der Haut exprimiert. Doch weder die genaue Zusammensetzung der CYP3A-Isozyme noch deren physiologische Rolle in der Haut sind bisher bekannt. Basierend auf begrenzten in vitro-Daten ist eine Rolle der CYP3A in der kutanen Vitamin D-Synthese denkbar. Auf der anderen Seite könnten die kutanen CYP3A auch lokal oder systemisch verabreichte Medikamente in der Haut verstoffwechseln und so zur Entstehung immunologischer und nicht-immunologischer unerwünschter Arzneimittelwirkungen beitragen, von denen sich bis zu 45 % in der Haut manifestieren.rnDie Arbeitshypothese dieses Projekts war, dass die CYP3A die kutane Synthese von Vitamin D regulieren. In dieser Funktion wurden sie zur Vermeidung von Vitamin D-Mangel-Erkrankungen wie Rachitis oder Osteomalazie in Europäern negativ selektiert. rnDie Expression und Regulation der CYP3A wurde in Hautbiopsien, einer Zelllinie epidermalen Ursprungs und primären Hautzellen wie auch in transgenen Mäusen untersucht. Die metabolische Aktivität der CYP3A gegenüber den kutanen Vitamin D-Vorstufen wurde mit Hilfe rekombinant exprimierter Enzyme untersucht. CYP3A5-mRNA war die häufigste der CYP3A in humanen Hautproben und überstieg die von CYP3A4 um das Dreifache, die von CYP3A7 um das 130-Fache. Damit entsprach diese 1,3 %, 0,01 % bzw. 0,01 % der jeweiligen hepatischen Genexpression. Die Expression von CYP3A43 war zu vernachlässigen. CYP3A5 zeigte eine bimodale Expression sowohl auf mRNA- als auch auf Proteinebene. So zeigten Träger der Wildtyp-Allels *1 eine 3,3-fach höhere mRNA- und 1,8-fach höhere Proteinmenge als homozygote Träger des Nullallels *3. CYP3A4/7- und CYP3A5-Protein wurde v. a. in den Keratinozyten der Epidermis und den Talgdrüsen, also den Bereichen der kutanen Vitamin D-Synthese lokalisiert. Die CYP3A5-Expression wurde ferner in der Haut transgener Mäusen gezeigt, die das Reportergen Luziferase unter Kontrolle des humanen CYP3A5-Promoters exprimieren. Verglichen mit der Leber war die kutane Expression des Vitamin D-Rezeptors (VDR) 100-fach höher, die der Xenosensoren CAR und PXR vergleichbar bzw. zu vernachlässigen. Dementsprechend erhöhte die Behandlung mit 1,25-Dihydroxyvitamin D, dem aktiven Vitamin D-Hormon, und dessen Vorstufen außer 7-Dehydrocholesterol, jedoch nicht der PXR-Ligand Rifampicin, die Expression der CYP3A. Wie in Zwei-Hybrid-Experimenten gezeigt, wurden die Effekte des 1,25-Dihydroxyvitamin D und dessen Vorstufen alleinig durch VDR vermittelt. Die Effektstärke hingegen war abhängig von Zellspender, Zellpassage und Zelltypus. Alle drei CYP3A-Isozyme metabolisieren Vitamin D zu einem oder mehreren unbekannten Metaboliten, jedoch nicht zu 25-Hydroxyvitamin D, dem direkten Vorläufer des aktiven Vitamin D. rnZusammengefasst legen die Daten nahe, dass die kutanen CYP3A, allen voran CYP3A5, die Vitamin D-Homöostase durch VDR-vermittelte Induktion des Abbaus von Vitamin D-Vorstufen regulieren. Dies zusammen mit Sequenzdaten liefert starke Indizien für Vitamin D als treibende Kraft der Selektion des CYP3A-Lokus in Europäern. Der Einfluss der CYP3A-Expression auf selektiv wirksame, klinisch relevante Knochenveränderungen wie Rachitis oder Osteomalazie müssen folgen.rn

Reducing fall risk in the elderly: risk factors and fall prevention, a systematic review

Falls in the elderly are a major source of injury resulting in disability and hospitalization. They have a significant impact on individual basis (loss of quality of live, nursing home admissions) and social basis (healthcare costs). Even though falls in the elderly are common there are some well studied risk factors. Special emphasis should be put on sarcopenia/frailty, polypharmacy, multimorbidity, vitamin D status and home hazards. There are several well evaluated fall prevention approaches that either target a single fall risk factor or focus on multiple risk factors. It has to be kept in mind that not all fall prevention strategies are useful for all patients as for example dietary substitution of vitamin D is only recommended in people with increased risk for a vitamin D deficiency. Home hazard reduction strategies are more effective when combined with other fall prevention approaches such as for example exercise programs. In conclusion elderly patients should routinely be screened for relevant risk factors and if need an indiviudally targeted fall prevention program compiled.

Evolutionary importance for the membrane enhancement of the production of vitamin D3 in the skin of poikilothermic animals.

The photoproduction of vitamin D in the skin was essential for the evolutionary development of terrestrial vertebrates. During exposure to sunlight, previtamin D3 formed in the skin is isomerized to vitamin D3 (calciol) by a temperature-dependent process. Since early land vertebrates were poikilothermic, the relatively slow conversion of previtamin D3 to vitamin D3 at ambient temperature put them at serious risk for developing vitamin D deficiency, thus leading to a poorly mineralized skeleton that could have ultimately halted further evolutionary development of vertebrates on land. We evaluated the rate of isomerization of previtamin D3 to vitamin D3 in the skin of iguanas and found the isomerization rate was enhanced by 1100% and 1700% at 25 degrees C and 5 degrees C, respectively. It is likely that the membrane entrapment of previtamin D3 in its s-cis,s-cis conformation is responsible for the markedly enhanced conversion of previtamin D3 to vitamin D3. The membrane-enhanced production of vitamin D3 ensures the critical supply of vitamin D3 to poikilothermic animals such as iguanas.

Concentrações séricas de 25 (OH) e perfil metabólico mediados pela adiposidade

Introdução: Baixas concentrações séricas de hidroxivitamina D (25[OH]D) e o excesso de peso atingiram níveis epidêmicos em todo o mundo. Estudos relatam que concentrações séricas de vitamina D estão associadas às alterações lipídicas, glicolíticas e inflamatórias; e estas alterações são conhecidamente mediadas pela adiposidade. Dessa forma, a vitamina D pode atuar de forma benéfica sobre o perfil metabólico em adolescentes, adultos e idosos. Objetivo: Investigar e descrever as associações entre as concentrações séricas de 25(OH)D e o perfil metabólico, mediadas pela adiposidade em adolescentes, adultos e idosos. Metodologia: Inicialmente, foi utilizada subamostra do Inquérito de Saúde de São Paulo (ISA-Capital), estudo transversal, de base populacional (n=281), para investigar a associação entre as concentrações séricas de vitamina D e marcadores inflamatórios em adultos brasileiros. Posteriormente, foram utilizados dados do estudo Healthy Lifestyle in Europe by Nutrition in Adolescents-(HELENA), estudo multicêntrico transversal da população de adolescentes européia, com o intuito de avaliar as alterações nos marcadores lipídicos e de homeostase da glicose mediados pela deficiência de vitamina D e obesidade. Finalmente, foi analisada a amostra do estudo PHYSMED, um estudo transversal com idosos não institucionalizados para verificar associações entre concentrações séricas de vitamina D, perfil lipídico e composição corporal em idosos espanhóis aparentemente saudáveis. Resultados: Nos adultos, observou-se uma associação negativa entre as concentrações de TNF-alfa e de IL-6 e as concentrações séricas de 25(OH)D em indivíduos com peso normal. Nos adolescentes, as concentrações de 25(OH)D foram associadas de forma independente e positiva com o Quantitative Insulin Sensitivity Check Index-QUICKI (p <0.001) e negativamente associada com o IMC (p <0.05). Também foi observado que o aumento do IMC esteve associado com um aumento de 1.93 vezes maior chance de deficiência de vitamina D (IC de 95 por cento = 1.03 - 3.62; p = 0.040). Em idosos, verificou-se que as concentrações séricas de 25(OH)D foram associadas com o IMC (p = 0.04), a circunferência da cintura (p = 0.004), CT/HDL-c (p = 0.026) e o HDL-c (p = 0.001). Adicionalmente, foi observado que idosos com concentrações de HDL-c <40mg/dl possuíam 1.7 vezes maior chance de apresentarem deficiência de vitamina D em comparação com aqueles que possuíam concentrações de HDL-c >40 mg/dl (95 por cento IC = 1.10 a 2.85; p = 0.017) e o aumento na circunferência da cintura também foi associado com um maior risco de deficiência de vitamina D (95 por cento IC =0.96-1.00; p = 0.04). Conclusão: A composição corporal interage com as concentrações de 25(OH)D modulando a resposta inflamatória, à homeostase da glicose e também o perfil lipídico. Indivíduos sem deficiência de vitamina D apresentam melhor perfil metabólico e também melhor composição, sugerindo que a suficiência de vitamina D pode ter um papel importante nas condições metabólicas mediadas pela adiposidade.

Season of birth is associated with anthropometric and neurocognitive outcomes during infancy and childhood in a general population birth cohort

The 'season of birth' effect is one of the most consistently replicated associations in schizophrenia epidemiology. In contrast, the association between season of birth and development in the general Population is relatively poorly understood. The aim of this study was to explore the impact of season of birth on various anthropometric and neurocognitive variables from birth to age seven in a large, community-based birth cohort. A sample of white singleton infants born after 37 weeks gestation (n =22,123) was drawn from the US Collaborative Perinatal Project. Anthropometric variables (weight, head circumference, length/height) and various measures of neurocognitive development, were assessed at birth, 8 months, 4 and 7 years of age. Compared to surnmer/autumn born infants, winter/spring born infants were significantly longer at birth, and at age seven were significantly heavier, taller and had larger head circumference. Winter/spring born infants were achieving significantly higher scores on the Bayley Motor Score at 8 months, the Graham-Ernhart Block Test at age 4, the Wechsler Intelligence Performance and Full Scale scores at age 7, but had significantly lower scores on the Bender-Gestalt Test at age 7 years. Winter/spring birth, while associated with an increased risk of schizophrenia, is generally associated with superior outcomes with respect to physical and cognitive development. (c) 2005 Elsevier B.V. All rights reserved.

Fibromyalgia and bipolar disorder: emerging epidemiological associations and shared pathophysiology

Fibromyalgia (FM) is a prevalent disorder defined by the presence of chronic widespread pain in association with fatigue, sleep disturbances and cognitive dysfunction. Recent studies indicate that bipolar spectrum disorders frequently co-occur in individuals with FM. Furthermore, shared pathophysiological mechanisms anticipate remarkable phenomenological similarities between FM and BD. A comprehensive search of the English literature was carried out in the Pubmed/MEDLINE database through May 10th, 2015 to identify unique references pertaining to the epidemiology and shared pathophysiology between FM and bipolar disorder (BD). Overlapping neural circuits may underpin parallel clinical manifestations of both disorders. Fibromyalgia and BD are both characterized by functional abnormalities in the hypothalamic-pituitary-adrenal axis, higher levels of inflammatory mediators, oxidative and nitrosative stress as well as mitochondrial dysfunction. An over-activation of the kynurenine pathway in both illnesses drives tryptophan away from the production of serotonin and melatonin, leading to affective symptoms, circadian rhythm disturbances and abnormalities in pain processing. In addition, both disorders are associated with impaired neuroplasticity (e.g., altered brain-derived neurotrophic factor signaling). The recognition of the symptomatic and pathophysiological overlapping between FM and bipolar spectrum disorders has relevant etiological, clinical and therapeutic implications that deserve future research consideration.

Actitudes hacia la Enfermedad del Alzheimer según el género y la edad de adultos colombianos

La enfermedad de Alzheimer (EA) es la demencia más frecuente y su prevalencia continúa en aumento tanto en Colombia como en el mundo. Esta investigación tuvo como objetivo explorar si las actitudes hacia la EA varían según la edad y género de 450 personas adultas colombianas. Se realizó un estudio exploratorio de corte transversal en el que se aplicó un cuestionario autodiligenciado. Se encontró que efectivamente hay algunas diferencias según la edad y el género en el componente cognoscitivo (creencias y conocimiento) y conductual (intención conductual y conducta) de las actitudes; y diferencias según el género en el componente afectivo. Se concluye que los conocimientos sobre la EA son escasos, que la tristeza es la emoción predominante hacia la EA y que es un tema de interés en el que predomina la idea de que afecta especialmente la memoria. Se discutieron los resultados reconociendo que esta es una aproximación inicial a las actitudes hacia la EA.

Deficiency in SLC25A1, encoding the mitochondrial citrate carrier, causes combined D-2- and L-2-hydroxyglutaric aciduria

The Krebs cycle is of fundamental importance for the generation of the energetic and molecular needs of both prokaryotic and eukaryotic cells. Both enantiomers of metabolite 2-hydroxyglutarate are directly linked to this pivotal biochemical pathway and are found elevated not only in several cancers, but also in different variants of the neurometabolic disease 2-hydroxyglutaric aciduria. Recently we showed that cancer-associated IDH2 germline mutations cause one variant of 2-hydroxyglutaric aciduria. Complementary to these findings, we now report recessive mutations in SLC25A1, the mitochondrial citrate carrier, in 12 out of 12 individuals with combined D-2- and L-2-hydroxyglutaric aciduria. Impaired mitochondrial citrate efflux, demonstrated by stable isotope labeling experiments and the absence of SLC25A1 in fibroblasts harboring certain mutations, suggest that SLC25A1 deficiency is pathogenic. Our results identify defects in SLC25A1 as a cause of combined D-2- and L-2-hydroxyglutaric aciduria.

Deficiency in parvalbumin, but not in calbindin D-28k upregulates mitochondrial volume and decreases smooth endoplasmic reticulum surface selectively in a peripheral, subplasmalemmal region in the soma of Purkinje cells

The Ca(2+)-binding proteins parvalbumin (PV) and calbindin D-28k (CB) are key players in the intracellular Ca(2+)-buffering in specific cells including neurons and have profound effects on spatiotemporal aspects of Ca(2+) transients. The previously observed increase in mitochondrial volume density in fast-twitch muscle of PV-/- mice is viewed as a specific compensation mechanism to maintain Ca(2+) homeostasis. Since cerebellar Purkinje cells (PC) are characterized by high expression levels of the Ca(2+) buffers PV and CB, the question was raised, whether homeostatic mechanisms are induced in PC lacking these buffers. Mitochondrial volume density, i.e. relative mitochondrial mass was increased by 40% in the soma of PV-/- PC. Upregulation of mitochondrial volume density was not homogenous throughout the soma, but was selectively restricted to a peripheral region of 1.5 microm width underneath the plasma membrane. Accompanied was a decreased surface of subplasmalemmal smooth endoplasmic reticulum (sPL-sER) in a shell of 0.5 microm thickness underneath the plasma membrane. These alterations were specific for the absence of the "slow-onset" buffer PV, since in CB-/- mice neither changes in peripheral mitochondria nor in sPL-sER were observed. This implicates that the morphological alterations are aimed to specifically substitute the function of the slow buffer PV. We propose a novel concept that homeostatic mechanisms of components involved in Ca(2+) homeostasis do not always occur at the level of similar or closely related molecules. Rather the cell attempts to restore spatiotemporal aspects of Ca(2+) signals prevailing in the undisturbed (wildtype) situation by subtly fine tuning existing components involved in the regulation of Ca(2+) fluxes.

Developmental Vitamin D-3 deficiency alters the adult rat brain

There is growing evidence that Vitamin D-3 (1,25-dihydroxyvitamin D-3) is involved in brain development. We have recently shown that the brains of newborn rats from Vitamin D-3 deficient dams were larger than controls, had increased cell proliferation, larger lateral ventricles, and reduced cortical thickness. Brains from these animals also had reduced expression of nerve growth factor (NGF) and glial cell line-derived neurotrophic factor. The aim of the current study was to examine if there were any permanent outcomes into adulthood when the offspring of Vitamin D-3 deficient dams were restored to a normal diet. The brains of adult rats were examined at 10 weeks of age after Vitamin D-3 deficiency until birth or weaning. Compared to controls animals that were exposed to transient early Vitamin D-3 deficiency had larger lateral ventricles, reduced NGF protein content, and reduced expression of a number genes involved in neuronal structure, i.e. neurofilament or MAP-2 or neurotransmission, i.e. GABA-(alpha 4). We conclude that transient early life hypovitaminosis D-3 not only disrupts brain development but leads to persistent changes in the adult brain. In light of the high incidence of hypovitammosis D-3 in women of child-bearing age, the public health implications of these findings warrant attention. (c) 2005 Elsevier Inc. All rights reserved.

High-density lipoprotein deficiency and dyslipoproteinemia associated with venous thrombosis in men

Background-Although dyslipoproteinemia is associated with arterial atherothrombosis, little is known about plasma lipoproteins in venous thrombosis patients. Methods and Results-We determined plasma lipoprotein subclass concentrations using nuclear magnetic resonance spectroscopy and antigenic levels of apolipoproteins AI and B in blood samples from 49 male venous thrombosis patients and matched controls aged <55 years. Venous thrombosis patients had significantly lower levels of HDL particles, large HDL particles, HDL cholesterol, and apolipoprotein AI and significantly higher levels of LDL particles and small LDL particles. The quartile-based odds ratios for decreased HDL particle and apolipoprotein AI levels in patients compared with controls were 6.5 and 6.0 (95% CI, 2.3 to 19 and 2.1 to 17), respectively. Odds ratios for apolipoprotein B/apolipoprotein AI ratio and LDL cholesterol/HDL cholesterol ratio were 6.3 and 2.7 (95% CI, 1.9 to 21 and 1.1 to 6.5), respectively. When polymorphisms in genes for hepatic lipase, endothelial lipase, and cholesteryl ester transfer protein were analyzed, patients differed significantly from controls in the allelic frequency for the TaqI B1/B2 polymorphism in cholesteryl ester transfer protein, consistent with the observed pattern of lower HDL and higher LDL. Conclusions-Venous thrombosis in men aged <55 years old is associated with dyslipoproteinemia involving lower levels of HDL particles, elevated levels of small LDL particles, and an elevated ratio of apolipoprotein B/apolipoprotein AI. This dyslipoproteinemia seems associated with a related cholesteryl ester transfer protein genotype difference. © 2005 American Heart Association, Inc.

Osteopenia in a teenage boy presenting with previously undiagnosed guanidinoacetate methyltransferase (GAMT) deficiency and response to creatine supplementation

A 16 y.o. fully ambulant boy born to consanguineous Indian parents, presented for assessment of a fragility femoral neck fracture sustained against a background of autism and moderately severe intellectual disability. He had a past history of infantile eczema, and epilepsy treated with anticonvulsants from 2 to 10 years of age, with no further seizures following cessation of anticonvulsants. He had a thin body habitus (see Table 1) with long fingers and a high arched palate. He had no speech and negligible social interaction, but physical examination was otherwise unremarkable. Positive investigations revealed an undetectable serum creatinine and a urinary metabolic screen which showed an elevated GUA:Phe of 160 (< 36) and a decreased creatinine of 0.3 mmol/l (1.2–29.5) consistent with the diagnosis of guanidinoacetate methyltransferase(GAMT) deficiency. He was commenced on oral creatine 5 g three times daily. Despite improvement in physical activity, height and bone density, there was no discernable improvement in his intellectual functioning. Proton and phosphorous brain and leg magnetic resonance spectroscopy(MRS) was performed at baseline and showed an increased inorganic phosphorus peak and decreased phosphocreatine synthesis in brain and decreased creatine concentration in muscle. Following creatine treatment total brain creatine(1H-MRS) and phosphocreatine/ATP ratio (31P-MRS) content increased to 30% and 60% of control values, respectively. Brain GUA returned to normal levels.

BRCA1 deficiency exacerbates estrogen-induced DNA damage and genomic instability

Germline mutations in BRCA1 predispose carriers to a high incidence of breast and ovarian cancers. BRCA1 functions to maintain genomic stability through critical roles in DNA repair, cell cycle arrest and transcriptional control. A major question has been why BRCA1 loss or mutation leads to tumors mainly in estrogen-regulated tissues, given that BRCA1 has essential functions in all cell types. Here we report that estrogen and estrogen metabolites can cause DNA double strand breaks (DSB) in estrogen receptor-α negative breast cells and that BRCA1 is required to repair these DSBs to prevent metabolite-induced genomic instability. We found that BRCA1 also regulates estrogen metabolism and metabolite-mediated DNA damage by repressing the transcription of estrogen-metabolising enzymes, such as CYP1A1, in breast cells. Lastly, we used a knock-in human cell model with a heterozygous BRCA1 pathogenic mutation to show how BRCA1 haploinsufficiency affects these processes. Our findings provide pivotal new insights into why BRCA1 mutation drives the formation of tumours in estrogen-regulated tissues, despite the general role of BRCA1 in DNA repair in all cell types.

Detection of nitrogen deficiency in wheat from spectral reflectance indices and basic crop eco-physiological concepts

We tested the capacity of several published multispectral indices to estimate the nitrogen nutrition of wheat canopies grown under different levels of water supply and plant density and derived a simple canopy reflectance index that is greatly independent of those factors. Planar domain geometry was used to account for mixed signals from the canopy and soil when the ground cover was low. A nitrogen stress index was developed, which adjusts shoot %N for plant biomass and area, thereby accounting for environmental conditions that affect growth, such as crop water status. The canopy chlorophyll content index (CCCi) and the modified spectral ratio planar index (mSRPi) could explain 68 and 69% of the observed variability in the nitrogen nutrition of the crop as early as Zadoks 33, irrespective of water status or ground cover. The CCCi was derived from the combination of 3 wavebands 670, 720 and 790 nm, and the mSRPi from 445, 705 and 750 nm, together with broader bands in the NIR and RED. The potential for their spatial application over large fields/paddocks is discussed.