dataesr/scientific-paragraphs-categorization huggingface dataset:
After this period, the cells were washed twice with PBS and the medium was replaced with fresh medium with 0. 5% FBS containing DHA at increasing concentrations (20-100 M) for a further 72 h. The number of viable cells exposed to DHA was evaluated by a colorimetric MTT assay. Data represent the mean of eight values and results and are expressed as viability in comparison with controls (100%). grow. Evidence in the literature suggests that n-3 PUFAs reduce the risk of BC, however these studies show that n-3 PUFA feeding was unable to mimic the phenotype observed in fat-1 mice  ( 31, 32 ). Expression of the fat-1 gene is a much more effective approach to modify fatty acid composition. By ubiquitously converting n-6 PUFAs to n-3 PUFAs, it signifi cantly increases the absolute level of n-3 PUFAs as well decreases the level of n-6 PUFAs leading to a decreased ratio of n-6/n-3 PUFAs in mouse tissues, which cannot be achieved by conventional dietary intervention.
This work was supported by a French Government grant managed by the French National Research Agency (ANR) under the program "Investissements d'Avenir" with reference ANR-11-LABX-0021-01-LipSTIC Labex. J. B. acknowledges support from the Région Bourgogne. This work was also partially supported by a grant from La Ligue contre le cancer and the Groupe Lipides et Nutrition (GLN). No potential confl icts of interest relevant to this article have been reported.
Using this approach we examined the permissiveness of the tet-P2 and tet-P2D alleles to tTa-mediated activation during different windows of time during development. In control experiments, olfactory epithelia from OMP-IRES-tTa/tet-P2 mice were examined at P30 for tet-P2 expression, as revealed by detection of GFP. We observed tet-P2-expressing neurons in the P2 zone at frequencies consistent with previous analyses (Figures  4D  and 2U , and data not shown). We next administered doxycycline-infused food to OMP-IRES-tTa/tet-P2 mice from embryonic day 0 (E0) to P5, via maternal feeding, and then maintained the animals without doxycycline to P30. In these mice we observed a dramatic decrease in the number of cells expressing tet-P2 and a basal shift in their distribution in the neuroepithelium (Figure  4E ).
To examine the possibility of biallelic OR expression, we generated a mouse line carrying the CaMKII-tTa transgene and compound heterozygous for the tet-P2 modification: one allele marked with GFP and the other with tau-lacZ (Figure  6B ). We analyzed expression of the tet-P2 alleles by immunohistochemical detection of GFP and lacZ in coronal sections of the olfactory epithelium of CaMKII-tTa/tet-P2/tet-P2Z animals. Both tet-P2 alleles were expressed in the epithelium at roughly equal frequencies, yet, remarkably, despite the genetic potential to express both, we observed that the vast majority of olfactory sensory neurons transcribed only one of the two tet-P2 alleles, with biallelic expression observed in only ,3% of the neurons (Figure  6E -6G and 6E9-6G9). We further analyzed these data by measuring the height of double (tet-P2+tet-P2Z+) and single positive (tet-P2+ or tet-P2Z+) neurons in the epithelium and observed a difference between the two populations: double positive neurons were found lower in the epithelium than the single positives, with mean relative positions (normalized to the height of the epithelium) of 0.
Stained sections were visualized, and wholemount visualization of endogenous GFP fluorescence was performed, with Zeiss 510 and 710 laser-scanning confocal microscopes. Relative cell position in the olfactory epithelium was determined by measuring the distance of the receptor positive neuron from the neuroepithelium-lamina propria interface divided by the basal-toapical height of the epithelium. Graph points represents an individual cell, with n = 100 for each genotype. All measurements were performed using Image J software, and graphs were created, and corresponding statistics performed, using GraphPad Prism 6. 0 software.
Rapid Amplification of cDNA Ends (RACE) and full-length cDNA cloning To isolate the rabbit full-length PMEL cDNA, we performed RACE experiments with 5 μg of total RNA extracted from dorsal skin of a Rex rabbit using the SMART RACE cDNA Amplification kit (Clontech-Takara Bio Europe, Saint-Germain-en-Laye, France), according to the manufacturer's instructions. Consensus primers (Table  1 ) were designed based on coding sequence homology between human and bovine PMEL gene (GenBank acc. no. NM_006928 and  Oulmouden et al. 2005). Nested PCR with consensus and adaptor primers was used for 5'-untranslated regions (UTR) and 3'-UTR amplification as previously described  (Fontanesi et al. 2010a , Table  1 ). PCR products were cloned into the pCR2. 1 vector (Invitrogen, Carlsbad, CA, USA) and sequenced as described below.
The amplified fragments were sequenced after treatment with 1 μL of ExoSAP-IT® (USB Corporation, Cleveland, Ohio, USA) for 15 min at 37 °C. Cycle sequencing of the treated PCR products was produced using the same PCR primers and the Big Dye v3. 1 kit (Applied Biosystems, Foster City, CA, USA). Sequencing reactions, after a few purification steps using EDTA, Ethanol 100 % and Ethanol 70 %, were loaded on an ABI3100 Avant capillary sequencer (Applied Biosystems, Foster City, CA, USA). Sequences were edited and aligned with the help of the CodonCode Aligner software (CodonCode Corporation, Dedham, MA, USA). PCR-RFLP (Table  1 ) was applied to genotype the g. 39510357G>A SNP (nomenclature of intronic SNPs is based on system coordinates of rabbit chromosome 4 in the oryCun2. 0 genome version: http://www.
Results and discussion Rabbit PMEL cDNA sequences The rabbit PMEL cDNA sequence obtained from the Rex rabbit included 29 bp of the 5'-UTR, 1 974 bp of the coding sequence (CDS) and 131 of the 3'-UTR (with the polyA tail), for a total of 2 135 bp (GenBank/EMBL acc. no. : FR849710). The coding sequence encoded for a predicted 657 amino acid sequence, that was 79 %, 82 %, 80 % and 78 % identical to the human (661 amino acids, P40967), horse (662 amino acids, NP_001157361), pig (636 amino acids, BAE03309) and cattle (649 amino acids, Q06154) PMEL amino acid sequences, respectively. As expected, the predicted rabbit PMEL amino acid sequence has the characteristic signature for mammalian PMEL protein  (Theos et al. 2005). A short N-terminal signal sequence of 24 residues is followed by a N-terminal region (NTR) of 200 amino acids that contains three putative N-terminal glycosylation sites and three cysteine residues.
Aligning the Rex rabbit cDNA sequence with that obtained from Angora and Chinchilla rabbits (GenBank/EMBL acc. no. : FR849711 and FR849709) and the rabbit cDNA sequence XM_002711060 (retrieved from GenBank and derived by automated computational analysis with gene prediction methods of sequencing data of the rabbit genome version oryCun2. 0, that, in the meantime, has been completed at the level of ~7X by the Broad Institute) we identified, on the whole, 10 SNPs, 8 of which are missense polymorphisms (c. 557T>G, p. V186G; c. 584A>G, p. Q195R; c. 603T>C; c. 854C>T, p. P285L; c. 878C>A, p. P293Q; c. 1016A>T, p. Q339L; c. 1256C>T, p. T419M; c. 1346G>T, p. S449I; c. 1501G>A, p. G501R; c. 1770T>C), and 2 are synonymous SNPs (c. 603T>C; c. 1770T>C).
The concentration of RNA was quantified by measuring absorbance at 260 nm (Multiskan Spectrum, Thermo Fischer Scientific, Waltham, MA, USA), and RNA integrity was checked using an Agilent 2100 bioanalyser (Agilent Technologies). The primers were designed using Primer Express Software (Applied Biosystems, France) based on sus scrofa published nucleotides sequences (Icare) and are described in Table  2. Reverse transcription was performed with 2 mg of total DNAse-treated RNA (DNase treatment performed by an Invitrogen kit, France). Real-time PCR was carried out on an ABI PRISM 7000 SDS thermal cycler (Applied Biosystems) by mixing 25 ml of PCR buffer (SYBRGreenTM PCR Master Mix; Applied Biosystems) with 500 nM of each primer, 5 ml of optimized concentration of the RT reaction product and 2 U of Uracyl DNA Glycosylase (Invitrogen).
3 Values were calculated according to  Sauvant et al. (2004). 4  Values were calculated according to  Noblet et al. (1994). Effect of extruded linseed in the sow diet on piglet fatty acid composition different tissues, but not on the other classes of fatty acids. Data were analysed by analysis of variance using the General Linear Model procedure of SAS  (SAS Institute Inc. , 1989). For zootechnical data, the analysis of variance tested the maternal diet effect. For the other parameters (milk, piglet tissues from days 0 to 28), we used the following model: g ðiÞ ¼ m þ a DðiÞ þ b AðiÞ þ g DÂAðiÞ þ U sowðiÞ þ ðiÞ where g (i ) is the dependent variable, m the overall mean, a D(i ) the systemic effect of the maternal diet, b A(i ) is the systemic effect of the day of lactation (for milk) or the piglet age (for piglet tissues) and g D 3 A(i ) the systemic effect of interaction between maternal diet and the day of lactation or piglet age.
Subjects Between 09/11/2006 and 09/03/2008, oxidative stress was measured on blood samples in our pediatric CF unit. We retrospectively analysed blood samples of 44 CF children collected during this period. Children were assessed once (n=19), twice (n=22) or three times (n=3), resulting in a total of 72 samples. During this out-clinic visit, anthropometric parameters (weight, height, body mass index (BMI) z-score, height to weight (H/W) ratio) and respiratory status (congestion, exacerbation, forced expiratory volume in 1 second (FEV1) (%) and maximum midexpiratory flow (MMEF)  [16] ) were noted in a computerized database. Their treatments, like fat-soluble vitamins, PERT, selenium supplementation, and ursodeoxycholic acid, were registered. A single physician (FB) also retrospectively evaluated CF severity scores (Shwachman and Brasfield scores).
The authors only found an improvement of plasma levels of some antioxidants (-carotene, CoQ-10, and -tocopherol). No correlation was found with urine 8-isoprostane levels. In recent reviews, details on antioxidant trials in CF were summarized  [1, 6]. Our data suggest that the KRL test is an interesting tool to assess antioxidant potential in CF. Moreover, its correlation with pulmonary status could dispense of the measure of BAL oxidative biomarkers that are more invasive. This test could allow to follow the antioxidant potential of CF children and to monitor the effectiveness of antioxidant intervention. In conclusion, the present study demonstrated that the whole antioxidant capability, measured by the KRL test, should be a useful biomarker to assess oxidative stress in CF patients. It was associated with pulmonary status.
Acknowledgements: Emmanuel Mas  was the recipient of a grant from  Lactalis  under the aegis of the  Groupe Francophone d'Hépatologie, Gastroentérologie et Nutrition Pédiatrique. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. AA: arachidonic acid, ALT: alanine amino-transferase, AST: aspartate aminotransferase, DHA: docosahexaenoic acid, EPA: eicosapentaenoic acid, GGT: gamma-glutamyl transpeptidase, ULN: upper limit of normal.
Beyond issues of hydration, previous studies have shown that plain water consumption was associated with higher quality diets, better health behaviors, and lower risk for chronic disease in youth and adults  [7, [14] [15] [16]. With some exceptions  [15, 17, 18] , few studies have explored the consumption of plain water among nationally representative samples of US adults. To our knowledge, no studies have examined water intake using the most recently available dietary data. In addition, previous work has not broken down water consumption by beverage category. The present study was conducted using a large and nationally representative database: National Health and Nutrition Examination Survey (NHANES) 2005-2010 for adults ≥20y. Estimates of total dietary water from all sources (including plain water) from other beverages and from moisture in foods were compared to the IOM AI values.
Dietary intake databases The present analyses used data from three cycles of the nationally representative National Health and Nutrition Examination Survey (NHANES), corresponding to years  2005-2006, 2007-2008 and 2009-2010. The National Center for Health Statistics (NCHS) has obtained IRB approval for all cycles of NHANES studies  [19]  and the data has been made available for public use via the NCHS website  [20]. The three NHANES cycles provided us with a nationally representative sample of 15,702 adults age ≥ 20y. These NHANES cycles were selected for two reasons. First, the collection of data on tap and bottled water consumed as a beverage only began in 2005 as part of the 24-h recall. In previous NHANES cycles, information about water as a beverage was not collected during the recall, but was assessed via questionnaire after the 24-h recall was complete.
For total water, water from all beverages and water from food sources, the model appropriate for ubiquitously consumed dietary components was employed. All other values were estimated using the episodic model. For example, tap water was consumed by only 67% of respondents while less than half the respondents consumed bottled water on their first recall. For no beverage type and in no subpopulation of interest did the frequency of consumption approach 90-95%, which would justify using the ubiquitous model. Additional covariates were included in the model to account for whether the recall data was from a weekday or weekend and whether it was the first or second recall  [25, 26]. In order to account for the complex survey design of NHANES data and estimate standard errors, balanced repeated replication (BRR) weights were constructed using WesVar software (Westat, Rockville, MD, 2012).
Family income-to-poverty ratios were defined as <1. 0, 1. 0-1. 99, 2. 0-3. 49, and ≥3. T-tests with unequal variances were used to test for differences in the mean intake level in each sub-group related to a reference group of interest. The reference groups used were age 20-50y, men, non-Hispanic whites, and those with a family income-to-poverty ratio ≥ 3. 5. All analysis used SAS software (Version 9. 4 of SAS System for Windows, SAS Institute, Cary, NC, 2013) and estimates of the usual intake distribution used macros, code and methods adapted from NCI and the Centers for Disease Control (CDC)  [28, 29].
Discussion These analyses of total water intakes from all sources, including tap and bottled water, were conducted among a representative sample of US adults from the NHANES 2005-2010 database. The amounts of dietary water provided by plain water and by other beverages and foods were then compared to AI values by gender and by age group. The intent was to examine how close the population came to meeting the AI values, as defined by the IOM DRIs. According to the IOM, AI values may be used as goals for individual intakes though there is much inter-individual variation for water needs. Health status, physical activity or strenuous work, and environmental factors, such as temperature and humidity, are additional aspects to be considered when evaluating adequate intakes at the individual level  [2, 13, [30] [31] [32].
Biological markers, including serum or plasma osmolality, and to some extent urine osmolality, are additional markers of hydration status  [10, 33, 34]. Until recently, hydration biomarkers were not included in NHANES data. In the 2009-2010 cycle, urine osmolality data were collected. Upon release of additional cycles that collect this data (to increase the sample size and availability of samples taken early in the morning), future work could examine patterns of this variable by population sub-group. However, no established cutoffs for hydration adequacy *Relative standard error is greater than 30%. 1 Milk consumed with food is included as a beverage. The present analyses of the observed water intakes relative to the indices of hydration suggest that water consumption ought to be monitored more closely  [35].
Another recent report, based on the 2007 National Cancer Institute's Food Attitudes and Behaviors Survey, examined behaviors and attitudes associated with low consumption of plain water among US adults  [48]. Here, the adjusted odds of drinking <4 cups of water per day was associated with older age (>55y), sedentary lifestyles and low consumption of fruits and vegetables, but not with education or incomes. However, in this study regular water consumption was reported via questionnaire, not by measurement in a dietary recall or interview. The present study therefore fills a gap in the existing knowledge regarding water consumption patterns among US adults. Although older adults are known to represent a group at risk, current data on water consumption patterns have not been available previously.
In the present analyses of NHANES 2005-2010 data, non-beverage food sources accounted for 17-25% of total dietary water, as compared to 19% reported in the 2010 Dietary Guidelines for Americans Advisory Committee report  [36] , though methods for assessing water intake have changed from previous NHANES cycles. This observation highlights that increasing consumption of low energy density foods with high water content foods (e. g. , fruits/ vegetables) is another approach to increase water intakes, while subsequently improving overall diet quality. The present analyses had some limitations. First, the NHANES data are based on self-report and are subject to random and systematic reporting errors. Each of the two dietary recall days used different methods to collect the data, which may introduce mode effects into the estimate of water consumption.
If water intakes were under-reported in the NHANES database, then the estimates presented here will over-estimate the percent of adults who fail to meet the recommended intakes. It is probable that many respondents under-reported water intakes due to drinking water lacking salience. This may be particularly problematic for events where little water was consumed or it was consumed casually (e. g. , repeatedly being refilled at a restaurant). It is important to note that these data cannot be directly compared to those from pervious cycles of NHANES (prior to 2005), as the mode for collecting data on water intake changed. In previous cycles of NHANES water intake was measured at the end of the recall via questionnaire, whereas in more recent cycles, water is measured as part of the 24hour recall.
Surveillance of water intake from dietary data should be carefully monitored and the use of biomarkers to evaluate hydration status at the population-level should be a priority. Nonetheless, the present analyses represent one of the few explorations of the consumption of water in the US and can be used to inform approaches to improving the overall diet quality and hydration status of the population. Advantages of the data used here include the use of a large and nationally representative dataset that forms the basis for dietary surveillance in the US.
in 1987  [40]  to a maximum of 24 months for the study by Di Iorio et al. in 2003  [41]. For this reason, the probabilities were standardised according to the duration of the follow-up period in the individual studies (Table  II ). Source: Foque and Laville (Cochrane) 2009 [26]   Table II also shows the data from the seven studies considered with regard to the probabilities and duration of follow-up-adjusted probabilities of observing the end-stage renal disease (ESRD) event or death for patients receiving/not receiving dietary treatment. The probabilities were adjusted by calculating for each one the respective annual rate from which we then obtained the annual probabilities  [42] [43]. Specifically the annual mortality or ESRD rate was calculated using the following formula:     t P r / 1 ln    Where P is the probability of the individual study considered and t is the amplitude of the range, expressed in years, that the probability considered refers to.
Acknowledgments This work was supported by  Wroclaw University of Environmental and Life Sciences  and by the  Institut National de la Recherche Agronomique. We would like to thank  Prof. Waldemar Rymowicz  from the  Department of Biotechnology and Food Microbiology, Wroclaw University of Environmental and Life Sciences  for access to the bioreactors. All authors have agreed to submit the manuscript to the '' Journal of Industrial Microbiology and Biotechnology ''.
Although a number of phosphate binders are in clinical development, none appears to constitute a significant step forward in phosphate control. However, nicotinamide (NAM, also known as niacinamide) may be a useful pharmacological alternative to binder-based approaches. Here, we review the data on NAM as a potentially valuable therapy for hyperphosphatemia. To this end, we searched the PubMed database with the keywords 'nicotinamide', 'niacin', 'hyperphosphatemia', 'chronic kidney disease', and 'phosphate binder', with a focus on the efficacy of NAM in dialysis patients.
Among the different signaling pathways, pERK1/2, pJNK and pAKT levels were decreased in MT2A-overexpressing cells compared with parental cells, whereas pP38 levels remained unchanged (Supplementary data 1A). A gene reporter assay confirmed that MT2A overexpression reduced transcriptional activity through AP1 complex (Supplementary data 1B). Finally, the pro-apoptotic ratio bax/bcl-2 was increased in MT2A-overexpressing cells compared with parental cells (Figure  2g ). Overall, these results suggest that the overexpression of MT2A in osteosarcoma cells leads to a reduction in cell proliferation rate and an induction of apoptosis, resulting in reduced cell viability. MT2A expression level influences osteoblastic differentiation in osteosarcoma cells. As proliferation and differentiation processes are usually interdependent, we evaluated the osteogenic potential of osteosarcoma cells modified for MT2A.
We found that MT2A overexpression induced an increase in mRNA levels of osteoblastic markers such as runx2, osterix, type I collagen and alkaline phosphatase in both SaOS2 and U2OS cell lines (Supplementary data 2A). Alkaline phosphatase enzymatic activity was also upregulated in MT2A-overexpressing cells compared with parental cells, as evaluated by colorimetric assay or cytochemical staining (Supplementary data 2B and C). Matrix mineralization was increased more than twofold in MT2A-overexpressing cells compared with parental cells in both cell lines despite their different basal differentiation stage (Supplementary data 2D and E). These data indicate that MT2A level of expression influences osteosarcoma cell differentiation toward the osteogenic lineage. MT2A overexpression modulates osteosarcoma cell viability through zinc chelation.
Alkaline phosphatase enzymatic activity and matrix mineralization were decreased in MT2Asilenced cells compared with control cells (Supplementary data 3B-E, respectively), confirming that MT2A influences osteoblastic cell differentiation. We then evaluated the cell response to chemotherapy drugs containing platinum or not. MT2A silencing amplifies the inhibitory effect of all tested cytotoxic compounds on U2OS cell viability (Figure  6d ). Cytotoxic activity of doxorubicin correlates with the amount of drug incorporated into DNA. MT2A silencing results in an increased level of doxorubicin combined with DNA, whereas MT2A overexpression markedly reduced the amount of DNA-incorporated doxorubicin (Figure  6e ). An indirect evaluation was also performed by the measure of Hoechst 33342 amount, a nuclear stain used as a post treatment to doxorubicin.