ROLE OF VITAMIN-D3 AND RETINOIC ACID IN A HUMAN THP-1 MACROPHAGE MODEL OF MYCOBACTERIUM TUBERCULOSIS INFECTION

Mycobacterium tuberculosis (Mtb) replicates within the human macrophages and we investigated the activating effects of retinoic acid (RA) and vitamin D3 (VD) on macrophages in relation to the viability of Mtb. A combination of these vitamins (RAVD) enhanced the receptors on THP-1 macrophage (Mannose receptor and DC-SIGN) that increased mycobacterial uptake but inhibited thesubsequent intracellular growth of Mtb by inducing reactive oxygen species (ROS) and autophagy. RAVD also enhanced antigen presenting and homing receptors in THPs that suggested an activated phenotype for THPs following RAVD treatment. RAVD mediated activation was also associated with a marked phenotypic change in Mtb infected THPs that fused with adjacent cells to formmultinucleate giant cells (MNGCs). Typically MNGCs occurred over 30 days of in vitro culture and contained non-replicating persisting Mtb for as long as 60 days in culture. We propose that the RAVD mediated inhibition of replicating Mtb leading to persistence of non-replicating Mtb within THPs may provide a novel human macrophage model simulating formation of MNGCs in humanlungs.

Food Politics: How the Food Industry Influences Nutrition and Health, Revised and Expanded Edition

A review of Food Politics: How the Food Industry Influences Nutrition, and Health, Revised and Expanded Edition.

Neural tube defects in Mexican-Americans living on the US-Mexico border: The effects of folic acid and dietary folate

Neural tube defects (NTDs) are malformations of the developing brain and spinal cord; the most common are anencephaly and spina bifida. Evidence from many populations suggests that 50% of NTDs can be prevented through daily consumption of folic acid. A recent study has reported that folic acid may not protect populations of Mexican descent. This finding has serious implications for women living along the US-Mexico border. Not only is risk high in these Mexican American women compared with other US women; they also differ markedly in supplemental folic acid and dietary folate consumption, and in NTD-related risks (e.g., obesity, diabetes). This case-control study investigated whether folic acid supplements and dietary folate reduces NTDs in Mexican Americans. Cases included liveborn, stillborn, electively and spontaneously aborted NTD-affected fetuses and infants occurring in the 14-county Texas-Mexico border. Controls were randomly selected from unaffected live births, frequency matched to cases by hospital and year. An in-person interview of 110 case and 113 control mothers solicited data on folic acid supplements, dietary folate, and other covariates. Consumption of folic acid-containing vitamins before conception was only 5% for both case and control women. Taking vitamins the trimester before conception had no apparent effect, after adjusting for covariates [odds ratio (OR) = 1.0, 95% confidence interval (CI) = 0.3–3.4]. Combining folate from vitamins and diet showed a 20% risk reduction for women consuming at least 400 μg of folate daily [OR = 0.8, 95% CI = 0.5–1.5]; however, this estimate is statistically indistinguishable from the null. Although consistent with an inherent ineffectiveness of supplemental folic acid, that so few women consumed multivitamins during the critical time severely limited the assessment of folic acid in this population. A reduced folate response in Mexican descent women may be due to a genetic heterogeneity for metabolizing folate. Alternatively, folate intakes may be insufficient to overcome other underlying risk factors. In conclusion, determining whether folic acid reduces NTD risk in Mexican American women requires further study in populations with higher folic acid exposures. Meanwhile, we should pursue all recommended prevention strategies to reduce risk, including motivating Mexican American women of childbearing age to take folic acid routinely. ^

Antioxidant supplementation attenuates oxidative stress in patients undergoing coronary artery bypass graft surgery.

Ischemia-reperfusion has been reported to be associated with augmented oxidative stress in the course of surgery, which might be causally involved in the onset of atrial fibrillation (AF), the most common arrhythmia after cardiac surgery. We hypothesized that supplementation of antioxidants and n-3 polyunsaturated fatty acids (n-3 PUFAs) might lower the incidence of AF following coronary artery bypass graft (CABG) surgery. In the present study, by monitoring oxidative stress in the course of CABG surgery, we analyzed the efficacy of vitamins (ascorbic acid and α-tocopherol) and/or n-3 PUFAs (eicosapentaenoic acid and docosahexaenoic acid). Subjects (n = 75) were divided into 4 subgroups: control, vitamins, n-3 PUFAs, and a combination of vitamins and n-3 PUFAs. Fluorescent techniques were used to measure the antioxidative capacity, i.e. ability to inhibit oxidation. Total peroxides, endogenous peroxidase activity, and antibodies against oxidized LDL (oLAb) were used as serum oxidative stress biomarkers. Post-operative increase in oxidative stress was associated with the consumption of antioxidants and a simultaneous onset of AF. This was confirmed through an increased peroxide level and a decreased oLAb titer in control and n-3 PUFAs groups, indicating the binding of antibodies to oxidative modified epitopes. In both subgroups that were supplemented with vitamins, total peroxides decreased, and the maintenance of a constant IgG antibody titer was facilitated. However, treatment with vitamins or n-3 PUFAs was inefficient with respect to AF onset and its duration. We conclude that the administration of vitamins attenuates post-operative oxidative stress in the course of CABG surgery.

Screening, Identification and Preliminary Investigation of Target Transporters in Pregnancy Pathologies

Screening, Identification and Preliminary Investigation of Target Transporters in Pregnancy Pathologies. INTRODUCTION: Pre-eclampsia (PE), intrauterine growth restriction (IUGR) and gestational diabetes mellitus (GDM) are major sources of clinical morbidity and mortality in pregnant women worldwide. The mechanisms underlying these gestational diseases are complex and not yet fully understood, but one factor contributing to their development is impaired maternal-fetal nutrient transport. Therefore, we aimed to identify candidate membrane transporters involved in transplacental nutrient transfer associated with PE/IUGR or GDM. METHODS: Using in silico strategies, we analysed various gene expression data sets generated on different platforms focusing on solute carriers, ABC transporters and TRP channels in order to identify transporters that are differently expressed between patients and gestational age-matched controls. These bioinformatic analyses were combined with literature data to define a catalogue of target transporters that could be involved in the development of PE/IUGR or GDM. Transporters of interest were then analysed for gene expression using qRT-PCR in placental tissues of patients and controls. For validating the results on protein and functional level, we started to establish an in vitro assay using freshly isolated primary cytotrophoblast cells polarized on the Transwell® system. RESULTS: Using bioinformatics approaches, we initially identified 37 target membrane proteins which were mainly associated with the transport of amino acids, vitamins, and trace elements. At the current state of analysis, the amino acid transporters SLC7A7, SLC38A2, SLC38A5, and the thiamine transporter SLC19A3 showed significant differences in placental mRNA expression between controls and patients affected by PE and/or IUGR. Subsequent gene expression analysis in our in-house GDM placental tissue bank is still ongoing. CONCLUSIONS: Based on our in silico analyses, literature data and first follow-up in vitro validations, we were able to define potentially interesting candidate transporters implicated in PE/IUGR or GDM. To date, additional newly defined candidate targets are being analysed on mRNA level in PE/IUGR and GDM. Subsequent analyses on protein and functional level will reveal whether these targets could be of diagnostic or therapeutical interest in these pregnancy pathologies.

Endocannabinoid transport revisited.

Endocannabinoids are arachidonic acid-derived endogenous lipids that activate the endocannabinoid system which plays a major role in health and disease. The primary endocannabinoids are anandamide (AEA, N-arachidonoylethanolamine) and 2-arachidonoyl glycerol. While their biosynthesis and metabolism have been studied in detail, it remains unclear how endocannabinoids are transported across the cell membrane. In this review, we critically discuss the different models of endocannabinoid trafficking, focusing on AEA cellular uptake which is best studied. The evolution of the current knowledge obtained with different AEA transport inhibitors is reviewed and the confusions caused by the lack of their specificity discussed. A comparative summary of the most important AEA uptake inhibitors and the studies involving their use is provided. Based on a comprehensive literature analysis, we propose a model of facilitated AEA membrane transport followed by intracellular shuttling and sequestration. We conclude that novel and more specific probes will be essential to identify the missing targets involved in endocannabinoid membrane transport.

Serum folate, cobalamin, homocysteine and methylmalonic acid concentrations in pigs with acute, chronic or subclinical Lawsonia intracellularis infection.

Lawsonia intracellularis is the causative agent of porcine proliferative enteropathy. The clinical presentation can be acute (i.e. proliferative hemorrhagic enteropathy, PHE), chronic (i.e. porcine intestinal adenomatosis, PIA) or subclinical. In humans with chronic enteropathies, low serum folate (vitamin B(9)) and cobalamin (vitamin B(12)) concentrations have been associated with increased serum concentrations of homocysteine and methylmalonic acid (MMA), which reflect the availability of both vitamins at the cellular level. The aim of this study was to evaluate serum folate, cobalamin, homocysteine and MMA concentrations in serum samples from pigs with PHE, PIA or subclinical L. intracellularis infection, and in negative controls. Serum folate, cobalamin, homocysteine and MMA concentrations differed significantly among pigs in the PHE, PIA, subclinical and negative control groups. Serum folate concentrations in the PHE and PIA groups were lower than in the subclinical and negative control groups, while serum cobalamin concentrations were lower in the PIA group than in other groups. Serum concentrations of homocysteine were higher in the PHE, PIA and subclinical groups than in the negative control group. Serum concentrations of MMA were higher in the subclinical and PIA groups than in the control group. These data suggest that pigs infected with L. intracellularis have altered serum cobalamin, folate, homocysteine and MMA concentrations.

Antioxidants improve vascular function in children conceived by assisted reproductive technologies: A randomized double-blind placebo-controlled trial.

AIMS Children conceived by assisted reproductive technology (ART) display vascular dysfunction. Its underlying mechanism, potential reversibility and long-term consequences for cardiovascular risk are unknown. In mice, ART induces arterial hypertension and shortens the life span. These problems are related to decreased vascular endothelial nitric oxide synthase (eNOS) expression and nitric oxide (NO) synthesis. The aim of this study was to determine whether ART-induced vascular dysfunction in humans is related to a similar mechanism and potentially reversible. To this end we tested whether antioxidants improve endothelial function by scavenging free radicals and increasing NO bioavailability. METHODS AND RESULTS In this prospective double-blind placebo controlled study in 21 ART and 21 control children we assessed the effects of a four-week oral supplementation with antioxidant vitamins C (1 g) and E (400 IU) or placebo (allocation ratio 2:1) on flow-mediated vasodilation (FMD) of the brachial artery and pulmonary artery pressure (echocardiography) during high-altitude exposure (3454 m), a manoeuver known to facilitate the detection of pulmonary vascular dysfunction and to decrease NO bioavailability by stimulating oxidative stress. Antioxidant supplementation significantly increased plasma NO measured by ozone-based chemiluminescence (from 21.7 ± 7.9 to 26.9 ± 7.6 µM, p = 0.04) and FMD (from 7.0 ± 2.1 to 8.7 ± 2.0%, p = 0.004) and attenuated altitude-induced pulmonary hypertension (from 33 ± 8 to 28 ± 6 mm Hg, p = 0.028) in ART children, whereas it had no detectable effect in control children. CONCLUSIONS Antioxidant administration to ART children improved NO bioavailability and vascular responsiveness in the systemic and pulmonary circulation. Collectively, these findings indicate that in young individuals ART-induced vascular dysfunction is subject to redox regulation and reversible.

Micronutrient Supplementation after Biliopancreatic Diversion with Duodenal Switch in the Long Term

Background Malabsorptive bariatric surgery requires lifelongmicronutrientsupplementation.Basedontherecommendations, we assessed the number of adjustments of micronutrientsupplementationandtheprevalenceofvitaminandmineral deficiencies at a minimum follow-up of 5 years after biliopancreatic diversion with duodenal switch (BPD-DS). Methods Between October 2010 and December 2013, a total of 51 patients at a minimum follow-up of 5 years after BPDDS were invited for a clinical check-up with a nutritional blood screening test for vitamins and minerals. Results Forty-three of fifty-one patients (84.3 %) completed the blood sampling with a median follow-up of 71.2 (range 60–102) monthsafter BPD-DS. At that time,all patientswere supplemented with at least one multivitamin. However, 35 patients (81.4 %) showed either a vitamin or a mineral deficiencyoracombinationofit.Nineteenpatients(44.1%)were anemic,and17patients(39.5%)hadanirondeficiency.High deficiency rates for fat-soluble vitamins were also present in 23.2 % for vitamin A, in 76.7 % for vitamin D, in 7.0 % for vitamin E, and in 11.6 % for vitamin K. Conclusions Theresultsofourstudyshowthattheprevalence ofvitaminandmineraldeficienciesafterBPD-DSis81.4%at a minimum follow-up of 5 years. The initial prescription of micronutrientsupplementationandfurtheradjustmentsduring thefirstfollow-upwereinsufficient toavoidlong-term micronutrient deficiencies. Life-long monitoring of micronutrients at a specialized bariatric center and possibly a better micronutrient supplementation, is crucial to avoid a deficient micronutrient status at every stage after malabsorptive bariatric surgery

Importance of cholesterol and cholesterol transporters in the placental trophoblast during pregnancy

Membrane transporters are essential during pregnancy, being a core component of the exchange of nutrients, gases, and metabolic products between the mother and the developing fetus. Important compounds to be transported include vitamins and minerals, amino acids, glucose, as well as cholesterol. Cholesterol transport across the plasma membrane is mediated mainly by members of the ATP-binding cassette (ABC) transporter family. Cholesterol is present in every cell of the body, where it helps maintain the integrity of cell membranes and also plays an important role in cell signaling events. Cholesterol also acts as a precursor for the biosynthesis of steroids that include sex hormones, glucocorticoids, mineralcorticoids, as well as bile acids and oxysterols. Cholesterol transport is therefore crucial for a host of different physiological processes. The following chapter addresses the involvement and importance of ABC transporters in these different processes. The critical role that ABC transporters Play for a successful pregnancy outcome is highlighted by pathological processes that result malfunction of cholesterol transport during pregnancy. Avenues of future research are also described, which may help to further delineate the function and mechanism of action of ABC transporters.

Modification of mutagen sensitivity by chemopreventive agents {\it in vitro\/}

The clinical application of chemopreventive agents is expected to prevent the appearance of cancer by arresting carcinogenesis or reversing it in the precancerous stages. The hypothesis of the present investigations was that chemopreventive agents (retinoids and antioxidant vitamins) may counteract the clastogenic effects of bleomycin in vitro in both lymphoblastoid cell lines and primary lymphocyte cultures and that a similar phenomenon can be detected in lymphocytes from individuals treated with 13-cis-retinoic acid. The efficacy of 13-cis-retinoic acid, n-(4-hydroxyphenyl)-retinamide, ascorbic acid, n-acetyl-l-cysteine, alpha-tocopherol, and alpha-tocopherol-acid succinate was tested against bleomycin-induced chromosomal breakage.^ The results provided direct evidence of the concentration-related protective effects of these agents against bleomycin-induced clastogenicity in cultures of human lymphoblastoid cell lines in vitro. Similar anticlastogenic protection was demonstrated with 13-cis-retinoic acid, ascorbic acid, n-acetyl-l-cysteine, and alpha-tocopherol-acid succinate in primary lymphocyte cultures in vitro. The in vitro anticlastogenic effect of 13-cis-retinoic acid was also demonstrated in lymphocyte cultures from peripheral blood samples from patients treated with this retinoid.^ An important consideration is that the concentrations used in the present investigations are comparable to those achieved in clinical situations.^ The in vitro anticlastogenic effect of these retinoids and antioxidants may constitute an important element of their chemopreventive properties. The results corroborate the hypothesis that these compounds may be effective in clinical chemoprevention trials. The bleomycin-assay may also be used as a short-term test to evaluate the antimutagenic effects of various agents. ^

AN ESTIMATION OF IN UTERO DRUG EXPOSURE BY DETERMINANTS OF FREQUENCY AND DISTRIBUTION OF MATERNAL DRUG CONSUMPTION

Data derived from 1,194 gravidas presenting at the observation unit of a city/county hospital between October 11, 1979 through December 7, 1979 were evaluated with respect to the proportion ingesting drugs during pregnancy. The mean age of the mother at the time of the interview was 22.0 years; 43.0 percent were Black; 34.0 percent Latin-American, 21.0 percent White and 2.0 percent other; mean gravida was 2.5 pregnancies; mean parity was 1.0; and mean number of previous abortions was 0.34. Completed interview data was available for 1,119 gravida, corresponding urinalyses for 997 subjects. Ninety and one-tenth percent (90.1 percent) of the subjects reported ingestion of one or more drug preparation(s) (prescription, OTC, or substances used for recreational purposes) during pregnancy with a range of 0 to 11 substances and a mean of 2.7. Dietary supplements (vitamins and minerals) were most frequently reported followed by non-narcotic analgesics. Seventy-six and one tenth percent (76.1 percent) of the population reported consumption of prescription medication, 42.5 percent reported consumption of over-the-counter medications, 45.7 percent reported consumption of a substance for recreational purposes and 4.3 percent reported illicit consumption of a substance. For selected substances, no measurable difference was found between obtaining the information from the interview method or from a urinalysis assay. ^

Aplicación de la clasificación ATC al análisis de las prescripciones en DAMSU : I : los grupos de medicamentos que encabezan el ranking

Este es un estudio sobre utilización de medicamentos donde se analiza la evolución de la prescripción, en DAMSU de UNCuyo, de los 14 grupos terapéuticos (GT) de la clasificación ATC, durante 4 años consecutivos. Su objetivo fue determinar la prevalencia de las prescripciones en los 3 primeros niveles de la clasificación. Los datos fueron recolectados en los meses de abril, junio, setiembre y diciembre utilizando la metodología del DURG y procesados con un programa EPI INFO. Las comparaciones estadísticas fueron realizadas mediante la Prueba no paramétrica de los Signos. El ranking de GT fue constante pero el total de prescripciones disminuyó significativamente entre 2004 y 2007. Los GT del 1º nivel: S. Nervioso (N), S. cardiovascular (C), Digestivo y Metabolismo (A) y Músculo-esquelético (M), ocuparon, en orden decreciente, los cuatro primeros puestos del ranking durante los 4 años. De estos GT fueron analizados los subgrupos del 2º y 3º nivel. La prescripción de Psicolépticos + Psicoanalépticos superó a la de Analgésicos en el grupo N. En el grupo C los Agentes Antihipertensivos, y entre ellos los IECAs, encabezaron el ranking. Las vitaminas fueron las primeras en el GT A y el subgrupo de Antiinflamatorios y Antirreumáticos en el GT M. Se discuten estos resultados en función de la racionalidad de las prescripciones.

(Table 1) Retinol (vitamin A) and alpha-tocopherol (vitamin E) concentration in polar bear (Ursus maritimus) kidney

Vitamins A and E content of inner organs, among these the kidneys, are increasingly being used as an indicator of adverse effects caused to the organism by e.g. environmental contaminants. In general, only a renal sub sample is used for analyses, and it is thus essential to know which part of the organ to sample in order to get a representative value for this important biomarker. The aim here was to assess the distribution of vitamins A (retinol) and E (alpha-tocopherol) within the polar bear multireniculate kidney (i.e. polar vs. medial position) and also within the cortex vs. medulla of each separate renculi. The results showed no significant difference between the medial and polar renculi with regards to either retinol (p = 0.44) or alpha-tocopherol (p = 0.75). There were, however, significant differences between cortex and medulla for both vitamins (retinol, p = 0.0003; alpha-tocopherol, p<0.0001). The kidney cortex contained higher values of both vitamins than the medulla; on average 29% more retinol and 57% more alpha-tocopherol. Mean concentrations in the medulla was 2.7 mg/kg for retinol and 116 mg/kg for alpha-tocopherol, and in the cortex 3.5 mg/kg for retinol and 182 mg/kg for alpha-tocopherol. These results clearly indicate that one should take precautions when analyzing retinol and alpha-tocopherol in polar bear kidneys. Prior to analysis, the renculi should be separated into medulla and cortex. The results indicated no significant differences between renculi from different parts of the kidney.

Influence of liposoluble vitamin status on health parameter and physical fitness in European adolescents = Influencia del estado en vitaminas liposolubles en parámetros de salud y condición física en adolescentes europeos

La adolescencia es un período de crecimiento y desarrollo crítico e importante para la adquisición de hábitos saludables, en los que tanto la alimentación como la actividad física tienen un papel destacado. Junto con el primer año de vida, los requerimientos de energía y nutrientes son mayores que en cualquier otro periodo. Dentro de la nutrición, las vitaminas se ven involucradas en múltiples procesos celulares y tisulares, y sus deficiencias se vinculan a enfermedades crónicas degenerativas en la edad adulta como las cardiovasculares, cáncer, diabetes y osteoporosis, pero cuyos factores de riesgo se establecen a edades más tempranas. Las concentraciones sanguíneas de vitaminas están influenciadas en gran medida por la ingesta dietética, pero existen otros factores del individuo, entre los que cabe citar la composición corporal, la actividad física y condición física que, junto a la genética, podrían desempeñar un papel crucial. La presente memoria de Tesis Doctoral tiene como objetivo analizar el estado en vitaminas liposolubles y su relación con diversos factores de salud, entre los que destacan la composción corporal, hábitos dietéticos, actividad física y condición física en adolescentes Europeos. El trabajo está basado en los datos del estudio HELENA (“Healthy Lifestyle in Europe by Nutrition in Adolescence”). Se han analizado un total de 1089 adolescentes procedentes de diez ciudades en nueve paises europeos. Los principales resultados de este trabajo indican; a) La existencia de un estado deficiente en vitaminas liposolubles en adolescentes Europeos, especialmente de vitamina D, que alcanza valores del 80%. b) La estación del año, la latitud, el índice de masa corporal, la condición física, la ingesta de calcio dietético, los suplementos vitamínicos y la edad son las variables más relacionadas con el estado de vitamina D. c) A su vez, la capacidad cardiorrespiratoria puede predecir los niveles de vitamina D en los chicos, mientras que la fuerza muscular y masa magra parecen influir en los niveles de vitamina D en las chicas. La grasa corporal y el índice de masa corporal se correlaccionan negativamente con los niveles de vitamina D, especialmente en chicos. d) Un estado de vitamina D óptimo provoca una mejora de la masa ósea sólo cuando se tiene un nivel adecuado de actividad física. e) Se identifica la necesidad de establecer un consenso sobre los rangos aceptables y puntos de corte para las concentraciones sanguíneas de estas vitaminas en este grupo de población, ya que los actuales están extrapolados de la población adulta ABSTRACT Adolescence is a critical period of physiological growth and development as well as for the acquisition of healthy behaviors where both diet and physical activity play a major role. Apart from the first year of life, both energy and nutrient requirements are greatest during adolescence and the way to spend this energy by movement is also crucial. Vitamins are specifically involved in multiple cellular and tissue processes, and there is increasing evidence that deficiencies at these early ages could contribute to risk factors of chronic diseases like cardiovascular and cerebrovascular disease, cancer, diabetes and osteoporosis in adulthood, regardless data are scarce for younger ages. Vitamin concentrations are largely influenced by diet but other individual factors like body composition, physical activity or fitness together with genetics could play also an important role. The current thesis analyzes the liposoluble vitamin status in European adolescents and their relation with several health related factors, like body composción, dietary intake, physical activity and fitness. The work is based on data from the HELENA study ("Healthy Lifestyle in Europe by Nutrition in Adolescence"), for which a total of 1089 adolescents from ten different cities, in nine European countries were recruited. The main outcomes of this thesis are: a) There is a high liposoluble vitamin deficiency prevalence in European adolescents, specifically for vitamin D, which is presenting almost 80% of the adolescents. b) Season, latitude, BMI, fitness, dietary calcium intake, supplements intake and age are highly related to 25(OH)D concentrations found in European adolescents. c) Cardiorespiratory fitness may predict 25(OH)D concentrations in male adolescents, whereas upper limbs muscular strength and FFM may predict 25(OH)D concentrations in young females. Fat mass and BMI are inversely related to 25(OH)D concentrations, especially in males. d) The effect of 25(OH)D concentrations on bone mineral content in adolescents depends on physical activity levels. e) There is a need to establish a consensus on acceptable ranges and cut-offs of blood concentrations of these vitamins during adolescence, as currently they are extrapolated from adults.