Rémi Thomasson

Nicolas Vignier

Cécile Peccate

Nathalie Mougenot

Philippe Noirez

Antoine Muchir
email: a.muchir@institut-myologie.org

Cecile Peccate

Alteration of performance in a

Keywords: Emery-Dreifuss muscular dystrophy, lamin, nuclear envelope, LMNA, performance

teaching and research institutions in France or abroad, or from public or private research centers.

INTRODUCTION

LMNA mutations cause an autosomal dominant inherited form of Emery-Dreifuss muscular dystrophy (EDMD) [START_REF] Bonne | Mutations in the gene encoding lamin A/C cause autosomal dominant Emery-Dreifuss muscular dystrophy[END_REF]. EDMD is characterized by the clinical triad of joint contractures that begin in early childhood, slowly progressive muscle weakness and wasting initially in a humero-peroneal distribution that later extends to the scapular and pelvic girdle muscles, and cardiac involvement that manifests as poor exercise tolerance, and congestive heart failure [START_REF] Emery | Unusual type of benign X-linked muscular dystrophy[END_REF][START_REF] Sanna | Cardiac features of Emery-Dreifuss muscular dystrophy caused by lamin A/C gene mutations[END_REF]. Clinical variability ranges from early onset with severe presentation in childhood to late onset with slow progression in adulthood. Cardiac involvement usually occurs after the second decade [START_REF] Captur | Lamin and the heart[END_REF]. LMNA encodes the A-type nuclear lamins [START_REF] Lin | Structural organization of the human gene encoding nuclear lamin A and nuclear lamin C[END_REF], which arise from alternative RNA splicing and are the constituents of nuclear lamina. Given the ability of mouse models to phenocopy various aspects of cardiovascular diseases, the use of mouse models has become critical to the study of cardiac biology, physiology, and novel therapeutics prior to translating findings to human. We largely contributed in understanding the metabolic and physiological mechanisms regulating cardiac function and energy balance in EDMD, both in animal models and in patient's tissues. These include abnormal regulation of the mitogenactivated protein kinase (MAPK) signaling cascade [START_REF] Muchir | Activation of MAPK pathways links LMNA mutations to cardiomyopathy in Emery-Dreifuss muscular dystrophy[END_REF][START_REF] Muchir | Abnormal p38alpha mitogen-activated protein kinase signaling in dilated cardiomyopathy caused by lamin A/C gene mutation[END_REF], altered actin dynamics [START_REF] Chatzifrangkeskou | Cofilin-1 phosphorylation catalyzed by ERK1/2 alters cardiac actin dynamics in dilated cardiomyopathy caused by lamin A/C gene mutation[END_REF], altered Wnt/βcatenin signaling [START_REF] Le Dour | Decreased Wnt/b-catenin signaling contributes to the pathogenesis of dilated cardiomyopathy caused by mutations in the lamin A/C gene[END_REF], fibrosis [START_REF] Chatzifrangkeskou | ERK1/2 directly acts on CTGF/CCN2 expresssion to mediate fibrosis in cardiomyopathy caused by mutations in the lamin A/C gene[END_REF], NAD signaling [START_REF] Vignier | Rescue of biosynthesis of nicotinamide adenine dinucleotide (NAD+) protects the heart in cardiomyopathy caused by lamin A/C gene mutation[END_REF] and AKT/mTOR signaling [START_REF] Choi | Temsirolimus activates autophagy and ameliorates cardiomyopathy caused by lamin A/C gene mutation[END_REF]. Despite the growing understanding of the pathogenesis of EDMD as well as the discovery of novel therapeutic for the disease [START_REF] Muchir | Abnormal p38alpha mitogen-activated protein kinase signaling in dilated cardiomyopathy caused by lamin A/C gene mutation[END_REF][START_REF] Le Dour | Decreased Wnt/b-catenin signaling contributes to the pathogenesis of dilated cardiomyopathy caused by mutations in the lamin A/C gene[END_REF][START_REF] Chatzifrangkeskou | ERK1/2 directly acts on CTGF/CCN2 expresssion to mediate fibrosis in cardiomyopathy caused by mutations in the lamin A/C gene[END_REF][START_REF] Vignier | Rescue of biosynthesis of nicotinamide adenine dinucleotide (NAD+) protects the heart in cardiomyopathy caused by lamin A/C gene mutation[END_REF][START_REF] Choi | Temsirolimus activates autophagy and ameliorates cardiomyopathy caused by lamin A/C gene mutation[END_REF][START_REF] Wu | Mitogen-activated protein kinase inhibitors improve heart function and prevent fibrosis in cardiomyopathy caused by mutation in lamin A/C gene[END_REF], little is known on the overall benefit of these treatments on the performance of the animals. There is a need for more comprehensive ways of assessing disease progression and treatment efficacy. Traditional approaches to monitoring disease progression in preclinical models of muscular dystrophies are limited mostly to histological analyses and functional measurements of single fibers or muscle groups. However, none of these tests can assess systemic muscle function in a non-invasive approach consistent with clinical assessment in EDMD patients.

Respiration is a product of systemic skeletal and cardiac muscle function [START_REF] Bassett | Limiting factors for maximum oxygen uptake and determinants of endurance performance[END_REF]. We here hypothesized that quantifying respiration during exercise would be a mean to studying systemic muscle function in a mouse model of EDMD, as it is commonly used to assess performance in humans [START_REF] Hoydal | Running speed and maximal oxygen uptake in rats and mice: practical implications for exercise training[END_REF]. During testing, cardiac output primarily drives the associated increase in oxygen consumption (VO 2 ) until peak oxygen consumption (VO 2 peak) and exhaustion is achieved. Combined measurements of exercise, respiratory performance and histological marker expression therefore provide a comprehensive, reliable, and powerful mean for assessing drug efficacy in a mouse model for EDMD, which in turn could help advance drug discovery.

RESULTS

Depressed left ventricular function in Lmna H222P/H222P mouse model

To explore the role of A-type lamins in the development of EDMD, we studied a mouse model that display a Lmna mutation changing the histidine in position 222 into a proline [START_REF] Hoydal | Running speed and maximal oxygen uptake in rats and mice: practical implications for exercise training[END_REF]. We used transthoracic echocardiography to determine the cardiac function of Lmna H222P/H222P male mice. While the cardiac structure and function were not statistically different at 3 months of age, at 6 months of age, Lmna H222P/H222P male mice have increased left ventricular diameters, and decreased fractional shortening (FS) compared with wild type animals (Figure 1A) [START_REF] Chatzifrangkeskou | Cofilin-1 phosphorylation catalyzed by ERK1/2 alters cardiac actin dynamics in dilated cardiomyopathy caused by lamin A/C gene mutation[END_REF][START_REF] Le Dour | Decreased Wnt/b-catenin signaling contributes to the pathogenesis of dilated cardiomyopathy caused by mutations in the lamin A/C gene[END_REF][START_REF] Arimura | Mouse model carrying H222P-Lmna mutation develops muscular dystrophy and dilated cardiomyopathy similar to human striated muscle laminopathies[END_REF]. Left ventricular dilatation in Lmna H222P/H222P male mice at 6 months of age was demonstrated by histopathological analysis (Figure 1B). M-mode transthoracic echocardiography showed increased left ventricular end-systolic and end-diastolic diameters in Lmna H222P/H222P mice compared with wild type mice (Figure 1B). We showed a marked increased of myocardial fibrosis in heart from Lmna H222P/H222P male mice at 6 months of age, as observed by Sirius Red staining (Figure 1C).

Alteration of body composition in Lmna H222P/H222P mouse model

As the EDMD progresses further, skeletal muscle continues to be lost, which can result in weight loss. We assessed the body composition using DEXA lean/fat in Lmna H222P/H222P mice. We observed that Lmna H222P/H222P mice have a significant difference in body weight compared with wild type mice (Figure 2A). In 3-month-old Lmna H222P/H222P mice, the mean of body weight was 24.12g, while from wild type mice it was 26.66g. At 6 months of age, the mean of body weight was 24.94g for Lmna H222P/H222P mice, while it was 28.14g for wild type mice. Further complicating these issues are the changes in body composition that are linked to EDMD. We observed that Lmna H222P/H222P mice have markedly increased body fat mass and relative body fat mass (Figure 2B, 2C) and reduced body lean mass and relative body lean mass (Figure 2B, 2C) compared with wild type mice. The loss of lean mass was consistent with the dystrophic process (Figure 3A, 3B) and the increase in adiposity could result from the decreased activity associated with the loss of activity.

Alteration of physical performance in Lmna H222P/H222P mouse model

We therefore asked whether we could assess the activity of Lmna H222P/H222P mice. Forced exercise on a treadmill was used to measure VO 2 peak. The VO 2 peak corresponds to both the cardiovascular system's functional limitation and the organism's aerobic capacity. Peak oxygen consumption has been previously validated to assess prognosis in patients with heart failure [START_REF] Chase | Effects of respiratory exchange ratio on the prognostic value of peak oxygen consumption and ventilator efficiency in patients with systolic heart failure[END_REF]. We showed that Lmna H222P/H222P mice have a significant reduction in VO 2 peak, as well as relative VO 2 peak, compared with wild type mice at 3 and 6 months of age (Figure 4A). The ratio VCO 2 /VO 2 is the respiratory exchange ratio (RER), which can be used to determine the proportion of carbohydrates and fatty acid utilized during an activity. Peak RER has been used as an objective parameter of effort [START_REF] Mezzani | Contribution of peak respiratory exchange ratio to peak VO2 prognostic reliability in patients with chronic heart failure and severely rduced exercise capacity[END_REF].

The Lmna H222P/H222P mice have a significant decreased RER at VO 2 peak (Figure 4B). The lower RER at VO 2 peak observed in Lmna H222P/H222P male mice indicates a higher level of lipid metabolism when compared with wild type mice. Importantly, there was a significant difference in the maximal speed and the running distance between the two genotypes (Figure 5); thus, the decreased metabolic performance of Lmna H222P/H222P mice resulted to a change in physical activity.

We next tested whether interventions on mice could alter these parameters of effort. Given that the NAD + precursor -nicotinamide riboside (NR) supplemented diet was recently shown to have benefit on cardiac contractility in Lmna H222P/H222P mice (11), we next assessed in a pilot study the NR effect on physical and metabolic performances in Lmna H222P/H222P male mice. Since we recently reported an aberrant cardiac NAD + content, we first assessed the steady-state skeletal muscle (soleus) NAD + level in both male wild type and Lmna H222P/H222P mice. We showed that the NAD + content was significantly lowered in hearts from Lmna H222P/H222P mice at 6 months of age compared with wild type mice, when the structure of soleus was altered (symptomatic) (Figure 6A). We next assessed the expression level of enzymes from the salvage pathway of NAD + biosynthesis by real-time PCR using RNA extracted from Lmna H222P/H222P mouse soleus at 6 months of age. There was a significant increased Nmrk2 mRNA expression in the soleus of Lmna H222P/H222P mice compared with wild type mice (Figure 6B). We showed that feeding Lmna H222P/H222P mice with NR, the substrate for Nmrk2, for 2 months improved significantly RER at VO 2 peak compared with chow diet-fed mice (Figure 6C).

We also reported a significant difference in the maximal speed and the distance between the NR-fed and the chow diet-fed groups (Figure 6D). Taken together these results showed, that RER at VO 2 peak as well as physical performance are reflecting a partially restoration of the striated muscle structure and function and could therefore be used to assess future benefit of treatment in mouse model of EDMD.

DISCUSSION

There is an important need for a more comprehensive approach to evaluate the efficacy of therapeutics for neuromuscular diseases. Although traditional measures of disease pathology are useful, they have a limited functionality given that they do not assess the whole body. Muscle disease is usually assessed by histological analysis and circulating markers of myofiber injury or functional assessments of isolated muscles. Our findings strongly suggest that non-invasive protocols for assessing exercise performance in Lmna H222P/H222P mice quantifying several parameters of respiration and exercise are useful to assess the dystrophic pattern in EDMD. Exercise performance (e.g. 6 minutes walk-test) was recently used as the primary endpoint to assess the benefit of p38 inhibitor ARRY-797 [START_REF] Muchir | Abnormal p38alpha mitogen-activated protein kinase signaling in dilated cardiomyopathy caused by lamin A/C gene mutation[END_REF] in patients with cardiomyopathy caused by LMNA mutation (ClinicalTrials.gov Identifier: NCT02057341). This clinical trial showed that patients receiving ARRY-797 performed better, with a mean change from baseline, of 69 meters.

Measures of respiratory performance during exercise have been established for other animal model of muscular dystrophy [START_REF] Rocco | A novel protocol for assessing exercise performance and dystropathophysiology in the mdx mouse[END_REF], but are lacking for mouse models of EDMD mice. We showed that respiratory performance was impaired in Lmna H222P/H222P mice. Our findings illustrate the usefulness of respiratory performance in quantifying the dystrophic phenotype in EDMD, as differences in several exercise parameters (e.g., VO 2 peak, RER) were detected even before detection of dystrophic pattern by histological analysis. Thus, quantifying respiratory performance during exercise is in some ways a more sensitive means for assessing muscular dystrophy in a mouse model for EDMD. The unique patterns of VO 2 peak and RER in exercising Lmna H222P/H222P mice identified novel and very useful markers for assessing exercise performance. Our findings would benefit the investigational drugs that could correct loss of skeletal muscle and could ameliorate performance in EDMD. Along these lines, we showed that interventional treatment using nicotinamide riboside (NR) significantly improved RER at VO 2 peak in Lmna H222P/H222P mice.

The RER is defined as the CO 2 produced/O 2 consumed. This ratio increases with the exercise intensity and is commonly used to indirectly determine the relative contribution of carbohydrate and lipids to overall energy expenditure. In Lmna H222P/H222P mice, we showed a decreased RER compared with wild type mice, indicating greater reliance on fatty acid oxidation. It has long been recognized that substrate metabolism is altered in the heart that undergoes pathological conditions [START_REF] Ritterhoff | Metabolism in cardiomyopathy: every substrate matters[END_REF]. We recently reported a marked decrease of pyruvate to lactate exchange kinetics in the heart muscle from Lmna H222P/H222P mice [START_REF] Cavallari | The 13 C hyperpolarized pyruvate generated by ParaHydrogen detects the response of the heart to altered metabolism in real time[END_REF]. Moreover, we recently reported a cardiac muscle nicotinamide adenine dinucleotide (NAD + ) depletion in Lmna H222P/H222P mice ( 9), suggesting a decreased activity of the pyruvate dehydrogenase complex and a reduced production of acetyl coenzyme A from carbohydrates. It would be interesting to extensively perform metabolomics studies in these mice to address the primary metabolic defect, which lead to decline in the function.

Assessing respiration (VO 2 and VCO 2 ) during exercise greatly increases the overall understanding of disease pathology, as it examines the combination of cardiac and skeletal muscle functions. Taken together, our study suggests that the use of respiratory performance during exercise to quantify changes in the dystrophic phenotype is a good complement to histological markers. Indeed, it can provide non-invasive markers for assessing EDMD disease progression, which is not always reflected histologically. The phenotypic changes observed in Lmna H222P/H222P mice can now be used to guide future interventional therapies to assess their beneficial effect. by comparison to housekeeping mRNA.

NAD + concentration quantification

Fresh mouse tissues were homogenized and processed according to previous report [START_REF] Vignier | Rescue of biosynthesis of nicotinamide adenine dinucleotide (NAD+) protects the heart in cardiomyopathy caused by lamin A/C gene mutation[END_REF]. The concentration of soleus NAD + was determined using a spectrophotometric assay as described previously [START_REF] Vignier | Rescue of biosynthesis of nicotinamide adenine dinucleotide (NAD+) protects the heart in cardiomyopathy caused by lamin A/C gene mutation[END_REF]. Absorption measurements were carried out in 96-well plates by spectrophotometry (Tecan).

Weight and body composition

The weight of the mice and their body composition (lean mass and fat mass) were measured throughout the protocol period. For body composition we used a nuclear magnetic resonance system (LF90II, Bruker, Germany).

Evaluation of the physical performance

Each mouse was evaluated on a one-way treadmill equipped with an indirect calorimetry system (Phenomaster, TSE, Germany). The performance evaluation protocol consisted of increasing the speed by 0.01 m.s -1 every 15 seconds. Oxygen consumption (VO 2 ), carbon dioxide production (VCO 2 ) and respiratory exchange ratio (RER = VCO 2 /VO 2 ) were recorded every second. Oxygen consumption peak (VO 2 peak in ml.h -1 ) was set as the highest VO 2 value achieved during the test. We developed an algorithm based on the Savitzky-Golay principle [START_REF] Savitzky | Smoothing and Differentiation of Data by Simplified Least Squares Procedures[END_REF] to automate the analysis of calorimetric data by smoothing the noisy data. The maximum speed reached by each mouse was recorded. The maximum speed corresponds to the speed at which the mouse can no longer run on the treadmill. All protocols were conducted for each mouse until exhaustion. Exhaustion was defined as the time when the animal "preferred" to receive electric stimulation through the grid installed at the beginning of the treadmill rather than continue the race. The intensity of the electric stimulation was 1.2 mA during 1s, with 1s latency. Before each protocol, there was a 3-minute acclimatization on the treadmill at low speeds from 0.01 to 0.05 m.s -1 . All these performance measurements were realized in the "Performance & Metabolism in Mice" facility (PMM, EA7329, Paris Descartes University, Paris, France).

Statistics

Statistical analyses were performed using GraphPad Prism software. Statistical significance between groups of mice was analyzed with a corrected parametric test (Welch's t test), with a value of P<0.05 being considered significant. To validate results, we performed a non-parametric test (Wilcoxon-Mann-Whitney test). Values are represented as means ± standard errors of mean (SEM).

FIGURES Six Figures

Figure 1 .

 1 Figure 1. Alteration of left ventricular function in Lmna H222P/H222P mice (A) Graphs showing mean left ventricular end diastolic diameter (LVEDD), mean left ventricular end systolic diameter (LVESD) and fractional shortening (FS) in 3-month-old and 6-month-old male Lmna H222P/H222P (H222P) and wild type (WT) mice (n=6). Values for each individual mouse as well SEMs of means (bars) are shown. ** P<0.005, *** P<0.0005. (B) Histological analysis of heart sections stained with hematoxylin and eosin from Lmna H222P/H222P (H222P) and wild type (WT) mice. Scale bar: 1mm. Transthoracic M-mode echocardiographic tracings in Lmna H222P/H222P (H222P) and wild type (WT) mice.

Figure 2 .

 2 Figure 2. Alteration of body composition in Lmna H222P/H222P mice (A) Graphs showing mean body weight (BW, g) in 3-month-old and 6-month-old male Lmna H222P/H222P (H222P) (n=16 at 3 months of age and n=20 at 6 months of age) and wild type (WT) mice. Values for each individual mouse as well SEMs of means (bars) are shown. * P<0.05, *** P<0.0005. (B) Graphs showing mean body fat mass (g) and body fat mass relative to BW in 3-month-old and 6-month-old male Lmna H222P/H222P (H222P) (n=16 at 3 months of age and n=20 at 6 months of age) and wild type (WT) mice.

Figure 3 .

 3 Figure 3. Dystrophic skeletal muscles in Lmna H222P/H222P mice Hematoxylin-Eosin staining of cross sections of diaphragm (A) and soleus (B) from 3 and 6-month-old male Lmna H222P/H222P (H222P) and wild type (WT) mice.

Figure 4 .

 4 Figure 4. Alteration of maximal performance in Lmna H222P/H222P mice (A) Graphs showing oxygen consumption (VO 2 ) peak (ml/h) and VO 2 relative to lean mass (ml/h/g) in 3month-old and 6-month-old male Lmna H222P/H222P (H222P) (n=16 at 3 months of age and n=20 at 6 months of age) and wild type (WT) mice. Values for each individual mouse as well as means (histograms) and SEMs of means (bars) are shown. **** P<0.00005. (B) Graphs showing respiratory exchange ratio (RER) atVO 2 peak in 3-month-old and 6-month-old male Lmna H222P/H222P (H222P) (n=16 at 3 months of age and

Figure 5 .

 5 Figure 5. Alteration of physical performance in Lmna H222P/H222P mice Graphs showing mean maximal speed (m/s) and distance (m) in 3-month-old and 6-month-old male Lmna H222P/H222P (H222P) (n=16 at 3 months of age and n=20 at 6 months of age) and wild type (WT) mice. Values for each individual mouse as well SEMs of means (bars) are shown. * P<0.05, *** P<0.0005, **** P<0.00005.

Figure 6 .

 6 Figure 6. Nicotinamide riboside supplementation leads to improvement of performance in Lmna H222P/H222P mice (A) Quantification of NAD + content in soleus from wild type (WT) (n = 6) and Lmna H222P/H222P (n=6) mice at 3 and 6 months of age. *P<0.05. (B) Expression of Nmrk2 mRNA obtained by qPCR from wild type (WT) (n=5) and Lmna H222P/H222P (n=5) mice at 6 months of age. ***P-value < 0.0005. (C) Graphs showing respiratory exchange ratio (RER) at VO 2 peak in 6-month-old male chow diet-fed (n=9) or NR-fed Lmna H222P/H222P (H222P) (n=10). Values for each individual mouse as well as means (histograms) and SEMs of means (bars) are shown. **P<0.005, *** P<0.0005. (D) Graphs showing

ACKNOWLEDGEMENTS

This study was supported by funds from the Association Française contre les Myopathies, from the Institut National de la Santé et de la Recherche Médicale, Sorbonne Université and Agence Nationale de la Recherche (ANR-17-CE17-0015-03). The authors gratefully acknowledge ChromaDex for providing NR. P.M. was supported by Région Ile de France and by labex GR-Ex. The labex GR-Ex, reference ANR-11-LABX-0051 is funded by the program "Investissements d'avenir" of the French National Research Agency, (ANR-11-IDEX-0005-02).

COMPETING INTERESTS

All other authors have declared no conflicts of interest.

MATERIAL AND METHODS

Mice

Lmna H222P/H222P mice [START_REF] Arimura | Mouse model carrying H222P-Lmna mutation develops muscular dystrophy and dilated cardiomyopathy similar to human striated muscle laminopathies[END_REF] were fed chow and housed in a disease-free barrier facility at 12h/12h light/dark cycles. All animal experiments were approved by the French Ministry of Health at the Center for Research in Myology for the Care and Use of Experimental Animals. The animal experiments were performed according to the guidelines from Directive 2010/63/EU of the European Parliament on the protection of animals used for scientific purposes. Chow supplemented with 0.25% NR (400 mg/kg) (Chromadex), as previously reported [START_REF] Vignier | Rescue of biosynthesis of nicotinamide adenine dinucleotide (NAD+) protects the heart in cardiomyopathy caused by lamin A/C gene mutation[END_REF] was formulated by Safe (Scientific Animal Food and Engineering).

Echocardiography

Mice were anesthetized with 0.75% isoflurane in O 2 and placed in a heating pad. Transthoracic echocardiography was performed using an ACUSON 128XP/10 ultrasound with an 11 MHz transducer applied to the chest wall. All parameters were measured by a "blinded echocardiographer" unaware of the genotype or treatment of the mice, performed in 2D mode and M-mode in triplicate.

Histology

Frozen pieces of soleus and diaphragm were mounted in Tissue-Tek (Fisher Scientific) and 10 µm sections cut on a cryostat. Sections were stained with hematoxylin and eosin for histological analysis.

Representa-tive sections were photographed using a Microphot SA (Nikon) light microscope attached to a Spot RT Slide camera (Diagnostic Instruments). Images were processed using Adobe Photoshop CS (Adobe Systems).

qPCR analysis

Total RNA was extracted using the RNeasy Mini kit (Qiagen). cDNA was synthesized using the SuperScript III synthesis system according to the manufacturer's instructions (Invitrogen). Real-time qPCR reactions were performed with SYBR Green I Master mix (Roche) using with the LightCycler


Faiza Hallouz
email: hallouzfaiza@gmail.com

Hafsa Karahacane

Mohamed Meddi

Yu Fisjak

Fadhila Sadi

Hanane Benchikh

Faiza Kemmerchou

Gil Mahe

Á H Karahacane

Á M Meddi

Á F Kemmerchou

Impact of Climate Variability on Hydrology of the Western Mitidja Watershed, Algeria

teaching and research institutions in France or abroad, or from public or private research centers.

Impact of Climate Variability on Hydrology of the Western Mitidja Watershed, Algeria 1 Introduction

Algeria is a country located in the North of Africa. Its climate is mainly semi-arid but also undergoes an important Mediterranean influence. Also, a change in rainfall is likely to lead to a change in the availability of water resources. The impacts on living things in these regions should therefore be significant. The western Mitidja basin risks, like many areas in Africa, being negatively affected by climate change. This motivated us to study the vulnerability of this basin, especially its surface waters, a fundamental element of development.

Materials and Methods

Study Area

The plain of Mitidja is located in the south of Algiers. It extends over the territories of Algiers, Boumerdes, Tipaza and Blida. With a WSW-ENE orientation and an average altitude of 100 m, it covers an area of 1300 km 2 . This region is split into two geographical areas, the East Mitidja and the West Mitidja. The Mitidja West, which constitutes our study area, receives a total rainfall of about 616.1 mm/year, of which 84.7% evaporates, 2.63% infiltrates and 12.67% flows.

Regionalization Using the Regional Vector Method

Then, we chose to carry out the regionalization using the regional vector method via the Hydraccess software, which objective is to reconstitute the missing rainfall data at the monthly and annual time step [START_REF] Singla | Impact du changement climatique global sur les régimes hydroclimatiques au Maroc tendances, ruptures et effets anthropiques sur les écoulements, mémoire de mastère2[END_REF].

Rupture Detection Tests on Time Series

A statistical approach was used to help us better analyze monthly and annual rainfall. Khronostat is a statistical analysis software of time series of which several tests of detection of ruptures are available. We have chosen those that allow us to determine ruptures dates: Pettit's nonparametric method, Bayesian method of Lee and Heghinian, and Hubert's segmentation [START_REF] Hallouz | Analyse des ruptures dans les séries pluviométriques dans le bassin de l'oued Mina (Nord Ouest d'Algérie)[END_REF]).

Modeling Rainfall Discharge

The model used in this study is the GR2M model (2-parameter Rural Model) which is a two-parameter rainfall-discharge model [START_REF] Makhlouf | Compléments sur le modèle pluie-débit GR4 J et essai d'estimation de ses paramètres[END_REF][START_REF] Perrin | Improvement of parsimonious model for stream flow simulation[END_REF].

Rainfall Data

The rainfall data come mainly from the database of the National Agency for Hydraulic Resources (ANRH). They are recorded from 23 rainfall stations spread over the catchment and whose size of observations varies from one station to another. Indeed, we divided the western Mitidja basin into two parts.

Runoff Data

The data bank for monthly flows comes from the National Agency for Hydraulic Resources (ANRH) of Blida.

The time series generally concern the period 1987-2012, but are of variable durations.

Results and Discussion

Regional Vector Results

It should be noted that in the dataset used for this study, the number of stations is high only during the period from 1973 to 2005. It can also be seen in both regions that the fluctuation of the annual rainfall index decreases with the number of stations observed per year. Indeed, 1982 and 1998, the indices of annual rainfall fluctuate with great amplitude and are then representative only of a small number of stations. And since 1979 until 1986, the observed stations are more numerous and the index of annual rainfall calculated by the regional vector is then more representative and has its variability reduced according to the years. This allows us to conclude that the amount of data constitutes a limit of the quality of annual rainfall indices of the regional vectors.

Rupture Results Interpretation and Analysis

The monthly and annual ruptures detected at rainfall stations show the rain evolution during the study period. A decrease in rainfall from the 70s was recorded at all study area stations. The deficit varies between 18.6 and 29.9%.

Indeed, a sharp decline at the stations studied since the 1970s and generally at the level of the western Mitidja basin. A decrease in flows was recorded on all the stations of the study area from the 70s.

Results of Calibration and Validation of the GR2M Model

The calibration of the model generally requires the use of some of the available data [START_REF] Refrsgaard | The role of distributed hydrological modeling in water resource management[END_REF].

Indeed, in order to determine the internal fit parameters of the model, the adopted calibration method, called "solver", shows that the hydrographs simulated by GR2M agree better with the hydrographs observed, which translates acceptable and superior Nash values to 60%. It can be said that the model simulation quality is good. Validation is an crucial part of the model development process. Like the calibration phase, we can establish the Nash criterion for each basin. The numerical result gave us Nash satisfactory 99, 9% for Boumedfaa and 100% for the Fer à cheval and Afroun stations.

Conclusion

The Mitidja West region is subject to a sub humid regional climate; its watershed contains many climatic and hydrological stations, although their distribution remains heterogeneous. The monthly and annual ruptures detected at these stations clearly show the evolution of the rain during the study period, a decrease in rainfall was recorded at all stations in the study area.

For the rainfall flow simulations, the efficiency of the model to simulate the monthly flows of the basin is given by the Nash criterion analysis, which has given satisfactory values greater than (60%). Therefore we can say that the GR2M model has a certain efficiency in simulating the flows at the monthly time step and to guarantee a better understanding of the hydrological behavior of the basin.

Finally, in order to evaluate whether the climate or man-made hydraulic developments were the cause of the decrease in flows, we compared the dates of the ruptures detected on the hydrological series at the dam construction dates of western Mitidja and the results obtained showed that it was more the decrease of rainfall that impacted the construction of the Bouroumi dam in 1985.


Gil Mahe
email: gil.mahe@ird.fr

Oula Amrouni
email: oulabz@yahoo.fr

Ben Thouraya
email: benmoussa.thouraya87@gmail.com

Laurent Moussa

Rajae El Dezileau
email: dezileau@gm.univ-montp2.fr

Hamadi Aoula

Abderraouf Habaieb
email: habaieb.hamadi@yahoo.fr

Ilias Hzami
email: iliaskacimi@yahoo.fr

Abderrahmane Kacimi

Fatma Khedimallah

Mohamed Kotti

Nadia Meddi
email: mmeddi@yahoo.fr

Saadi Mhammdi
email: nmhammdif@yahoo.com

Abdeljaouad

H Habaieb

A Khedimallah
email: a.khedimallah@ensh.dz

Á M Meddi

F Kotti
email: f.kotti@yahoo.fr

S Abdeljaouad
email: saadi_abdeljaouad@yahoo.fr

T B Moussa

L Dezileau

R El Aoula

Á N Mhammdi

Assessment of the Impact of Dams on River Regimes, Sediment Transports to the Sea, and Coastal Changes

Keywords: Rivers, North Africa, Dams, Sediment transport, Coastal morphological changes Sands

A great number of rivers of North Africa are equipped with many dams for multiple purposes, mainly potable water, irrigation and energy production. In most of the countries-Morocco, Algeria and Tunisia, the storage capacity exceeds the runoff capacity, which means that water is stored several times in a row during its course to the sea. This leads to the storage of massive amounts of sediments into the dams' lakes, but nothing is known about how much sediment is released from these dams. This topic is mainly unstudied by the regional scientists due to lack of measurements of sediment transport, or poor access to observed data when available. Thus there are no time series of sediment loads to the sea from the North African rivers. One of the conse-quences is that it is impossible to relate recently observed coastal morphological changes to changes in the sediment recharge from the continent. This study brings a synthesis of recent works on this topic over the largest rivers of North Africa in the three countries. In Morocco and Tunisia there no continuous time series of sediment transport observations. There we sampled sediment cores in river meanders, to draw the history of real sediment exportations to the sea, on the basis of granulometry, geochemical and isotopic studies. Only in Algeria are there observations of sediment transport at main hydrological stations. The times series show many lack of data which are reconstructed according to runoff/concentrations correlations, taking seasonal factor N.

into account. Methodologies and first results are shown in this study. They reflect the huge decrease of the sediment transport to the sea over time since several decades, and in the case of the Medjerda river in Tunisia, the absence of sands in the river sediments since 34 years and the construction of the major dam on the main stream. ). He is member of the pedagogical team of the Master GOES of the University Mohammed V of Rabat, and associate member of the Research Center on "Water, Natural Resources, Environment and Sustainable Development" of the University of Rabat. He is responsible for two research programs in Tunisia PHC/RYSCMED and in Morocco PHC/CASBA. He also participates in two research programs in Algeria and in Gabon/Congo. He has been elected member of the Specialists Commission no 36 at the University of Montpellier, and spent 13 years at IRD Specialists or partnership Commissions. He has been for 5 years team leader of the Water team of the IGCP/International Geological Program of UNESCO, and he is member of the Scientific Commission of the program "Abandonned Mines" of IGCP/Swedish funds. He has been member of the UNESCO expert group on the impact of land cover changes on hydrological regimes. He has been a 6-year Associated Editor of Hydrological Sciences Journal, and he is currently Associate Editor of Ecohydrology and Hydrobiology/Elsevier and of «Climatologie» AIC journal. He is responsible for the conference cycle on the "hydrology of African Large river basins" UNESCO/IAHS, started in 2015, and scientific editor of numerous special issues of journals and books. He is teaching since several years in Maghreb Universities and High Schools an initiation to bibliometry, to master and doctorate students and researchers.

Author Biography

  University on the topic of Relationships between man, environment and water resources in Africa and climate change. He received the "Georges Hachette Award" from the Société de Géographie de Paris for his Ph.D. work published by ORSTOM. He is responsible for the FRIEND program UNESCO. International Hydrological Program, and coordinator of the MEDFRIEND program for the Mediterranean. He is the President of the International Commission of Surface Water of the International Association of Hydrological Sciences, and member of the Scientific Commission of the International Sediment Initiative and of the Large River Initiative (UNESCO

	Dr. Gil Mahe Research Director at the
	Institut de Recherche pour le
	Développement (IRD). He is an hydro-
	climatologist specialized on West and
	Central Africa and Maghreb, Ph.D. of the
	Faculty of Sciences of Paris XI, on the
	topic of African discharges to the ocean
	and links with climate and ocean vari-
	ability, and HDR of the Montpellier


Issifou Yaya

Akouda Akessiwe Patassi

Essoya Dadja

Essodjelouna Manani Landoh

Kanfitine Bignandi

Abdel-Daim Kolani

Namoro Daou

P ' Niwe

Massoubayo Patchali

Didier Lihanimpo Djalogue

Koumavi Ekouevi

Professor Bayaki Saka

Akouda Akessiwè Patassi

Dadja Essoya Landoh

Essodjèlouna Manani Bignandi

Kanfitine Kolani

Abdel-Daim Daou

P ' Niwè

Didier Koumavi Ekouevi

Modern contraceptive use among HIV-infected women attending HIV care centres in Togo: a cross-sectional study

niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

bACkgrounD Although there has been a reduction in new infections of HIV of about 41% between 2000 and 2015, HIV infection continues to be a public health problem in Sub-Saharan Africa, where 25.6 million people lived with HIV. Women of reproductive age account for about two-thirds of them. [START_REF]HIV/AIDS[END_REF] The decline in the incidence of HIV infection is the effect of multiple interventions implemented to prevent HIV transmission among heterosexual and especially from mother-to-child HIV transmission. About 66 000 children under 14 years were reported to be newly infected by HIV in Western and Central Africa at the end of 2015. [START_REF]AIDS by the numbers[END_REF] Most of these infections could have been prevented by antiretroviral therapy (ART) during pregnancy and more by contraceptive use among HIV-infected women, which is the fourth component of preventing mother-to-child transmission (PMTCT). In 2005, Reynolds et al [START_REF] Reynolds | Contraception to prevent HIV-positive births: current contribution and potential cost savings in PEPFAR countries[END_REF] estimated that the level of contraceptive use could prevent over 173 000 unintended HIV-infected births each year in Sub-Saharan Africa.

Indeed, family planning showed numerous benefits for HIV-infected women, such as the reduction of morbidity and mortality due to pregnancy and also the improvement of health of HIV-infected women by reducing unintended pregnancies. Therefore family planning contributes to PMTCT of HIV. [START_REF] Crankshaw | Placing contraception at the centre of the HIV prevention agenda[END_REF] The strengths and limitations of this study ► The use of contraceptives such as condoms could provide dual protection against acquisition or transmission of sexually transmitted infections, including HIV. ► This study is one of the first studies in Togo focusing its interest on the contraceptive use among HIVpositive women. ► Education level, marital status, AIDS clinical stage, type of health centre and having children had influenced the use of contraceptive methods among HIVinfected women. ► It is suggested that caregivers should integrate family planning services in HIV care during follow-up visits. ► Finally, we relied on self-reported reproductive health behaviour through a face-to-face interview, which may introduce information bias.

Open Access use of contraceptives, mainly male and female condoms, could provide dual protection. It could protect against acquisition or transmission of sexually transmitted infections, including HIV. 5 6 However, in Sub-Saharan Africa, given the progress in PMTCT with the expansion of ART, most of the HIV-infected women still have desire to have children, 7 8 and the prevalence of contraceptive use varies according to the studies. In Western Africa, the use of contraceptive methods is not effective in HIV-infected women. [START_REF] Ogbe | Pattern of contraception among HIV positive women in Jos University Teaching Hospital[END_REF][START_REF] Ezugwu | Contraceptive use among HIVpositive women in Enugu, southeast Nigeria[END_REF][START_REF] Laryea | Contraceptive use and unmet need for family planning among HIV positive women on antiretroviral therapy in Kumasi, Ghana[END_REF] In Ghana, a study showed that only 42.6% of HIV-infected women used modern contraceptive methods at Komfo Anokye Teaching Hospital in 2012, [START_REF] Laryea | Contraceptive use and unmet need for family planning among HIV positive women on antiretroviral therapy in Kumasi, Ghana[END_REF] while in eastern Nigeria it was reported that 73.1% of HIV-infected women were using modern contraceptives in the same period. [START_REF] Ezugwu | Contraceptive use among HIVpositive women in Enugu, southeast Nigeria[END_REF] In many studies conducted in Sub-Saharan Africa, the reported factors influencing the use of contraceptive among HIV-positive women were sociodemographic characteristics (age, education, marital status), reproductive characteristics (desire to have children, number of children), being on ART and knowledge of the partner's HIV status. [START_REF] Ogbe | Pattern of contraception among HIV positive women in Jos University Teaching Hospital[END_REF][START_REF] Ezugwu | Contraceptive use among HIVpositive women in Enugu, southeast Nigeria[END_REF][START_REF] Laryea | Contraceptive use and unmet need for family planning among HIV positive women on antiretroviral therapy in Kumasi, Ghana[END_REF][START_REF] Kimani | Family planning use and fertility desires among women living with HIV in Kenya[END_REF][START_REF] Lawani | Dual method use for protection of pregnancy and disease prevention among HIV-infected women in South East Nigeria[END_REF][START_REF] Nieves | The influence of partnership on contraceptive use among HIV-infected women accessing antiretroviral therapy in rural Uganda[END_REF] In 2015, Togo had about 110 000 people living with HIV and AIDS (PLWHA), including 59 000 women aged more than 15 years old and 9000 children aged 0-14 years. [START_REF] Onusida Togo | Estimations VIH et SIDA[END_REF] In addition, the prevalence of HIV infection is almost twice higher among women than men (3.1% vs 1.7%). [START_REF]et de l'Aménagement du Territoire[END_REF] HIV prevalence among women attending antenatal consultation in Togo was 2% in 2015. [START_REF]Programme National de Lutte contre le Sida et le IST au Togo PNLS-IST[END_REF] In women of reproductive age, the prevalence of contraceptive use was estimated at 22% for all methods (16% for modern methods) in 2013-2014. [START_REF]et de l'Aménagement du Territoire[END_REF] However, contraceptive use among HIV-infected women in Togo is poorly documented.

This study aims at assessing the prevalence of contraceptive use, as well as individual and structural factors that influenced the use of contraceptives among HIV-infected women enrolled in HIV care centres in two health regions in Togo.

MethoDs study design

This was a cross-sectional study conducted in two health regions in Togo (Centrale and Kara regions) over a period of 4 months from May to August 2016. The study targeted HIV-positive women of reproductive age (15-49 years) and who were sexually active.

setting and study population Centrale and Kara regions are two of the six health regions in Togo located, respectively, about 350 km and 420 km from the capital Lomé. These two regions are characterised by important seasonal arrival of populations (from the other health regions, and sometimes from outside of the country (Togo)) during cultural and traditional events, increasing the risks of HIV transmission. In 2013, the prevalence of HIV infection was estimated at 2.2% in the Centrale region and 1.8% in the Kara region. [START_REF]et de l'Aménagement du Territoire[END_REF] There were 30 accredited medical centres in the two regions to provide healthcare to about 10 361 PLWHA, including 616 children, enrolled and followed up in these centres. [START_REF]Programme National de Lutte contre le Sida et le IST au Togo PNLS-IST[END_REF] Women were eligible to participate in the study if they (1) were aged between 15 and 49 years old, (2) had a sexual partner within the last 6 months and (3) were enrolled in the selected centres for active ART. sampling First, a random probability sampling proportional to the number of patients in the active file of the 30 HIV care centres in the two regions was carried out to ensure representative centres with a high number of PLWHA. In a minimalist scenario, it is assumed that a sample of 15% of HIV care centres should be representative of all the centres. This led to the random selection of five HIV care centres for the implementation of this study. Then, a non-probabilistic, convenience sampling was carried out. Indeed, in these five selected HIV care centres, it was proposed to enrol any PLWHA who consulted for a follow-up from May to August 2016, who met the inclusion criteria and who consented to participate in the study. The prevalence of the use of at least one contraceptive method was assumed to be 50% with the precision of 5% and 20% refusal or incomplete data. Based on this assumption, the sample size was estimated at 461 HIV-infected women.

Data collection

Data collection was done using a standardised questionnaire in French, explained in the local language for participants if needed. The questionnaires were filled by the health workers. The questionnaire included sociodemographic information, clinical features, information on ART, sexual activity status, and knowledge on contraceptives and their use. Data on HIV status disclosure to the sexual partner were also collected among PLWHA. We defined modern contraception as the use of one of the following methods: sterilisation, condoms, contraceptive pill/oral contraceptives, intrauterine device, diaphragm, injection, emergency contraception or implant. statistical analysis of data Data entry was performed using EpiData V.3.1 software. Data were then exported for statistical analyses using SPSS V.17.0.

For continuous variables, mean and SD were calculated, while for categorical variables we calculated the proportions. Our main outcome variable was HIV-infected women who were currently using any modern contraceptive methods. Pearson's χ 2 test or Fisher's exact test was used when appropriate in bivariate analysis. Multivariate backwards stepwise logistic regression analysis was performed to identify independent risk factors for the dichotomous outcome contraceptive use or not. All variables significant during bivariate analysis at a p value <0.20 were introduced in a logistic regression model to estimate the adjusted OR (aOR). Interactions between the independent variables were tested. All these analyses Open Access were performed with 95% CI. To verify the robustness of the results, a sensitivity analysis was conducted to confirm the results found. We have performed the analysis by including women who have never heard about contraceptive methods, and then we excluded them to appreciate the variability of the statistical associations. We did not observe significant variability and logically we excluded those women (15/461) from the model. Our final model included only HIV-infected women who had heard about contraceptives methods.

ethical issues

We obtained consent from patients who participated in the study. For each of the person surveyed, the objectives, benefits to participate in the survey and progress of the investigation were clearly stated, as well as their right to interrupt the interview without justification. An informed consent form signed after the verbal explanation was made by the investigating officer in the language understood by the participant.

results sociodemographic and clinical characteristics

Table 1 shows the sociodemographic and clinical characteristics of the participants. In total 461 HIV-infected women of reproductive age and who were sexually active were enrolled into this study, of whom 267 (57.9%) were in the Centrale region and 194 (42.1%) in the Kara region. The mean (±SD) age of the participants was 34.3±7.1 years, ranging from 16 to 49 years. Among them, 40.6% had primary education or higher education level (45.3%), 54.7% were living in couple and 62.3% were living in an urban area. Out of the 461 women interviewed, 415 (90.0%) were on ART and for more than 2 years for 71% of them. The mean (±SD) duration on ART was 4.1±2.8 years. A quarter (25.4%) of the participants had a CD4 cell count of <0.35 × 10 9 /L at the last visit. Patients were classified as stage I (55.0%) or stage II (27.3%) based on the WHO clinical classification at the time of the survey. The partner's HIV status was unknown for 56.4% (260/461) of the participants. Most of the patients were followed up in a public hospital centre (61.8%), in a centre with available doctor (86.3%) or with a psychologist (55.7%).

reproductive health outcome

The majority of women (85.3%) reported having at least one child, while six out of ten participants expressed desire of having kids. Four hundred and forty six (96.7%) women reported that they heard about contraceptives methods. Among them, 70.2% (313/446) were informed by healthcare providers. At the time of the survey, the proportion of women who were using a contraceptive method was 74.7% (333/446) (table 1), and among them 74.5% (248/333) reported using condom, 8.4% (28/333) were using hormonal contraceptive and 16.9% (56/333) reported using dual methods combining condom and hormonal contraceptive. For 15 (3.3%) women who have never heard about contraceptives methods, we considered they had missing data for the variable contraceptive use.

Furthermore the proportion of women using contraceptive was significantly higher among participants with higher level of education, ranging from 57.1% in women with no education to 82.8% in those having reached the secondary education level or higher (p<0.001). However, this proportion was significantly higher (p<0.001) among women at stage I (82.1%) than those at stage II (70.0%) or those at stages III and IV (58.4%). The prevalence of contraceptive use was significantly lower in women living in couple (p=0.002), higher among those who had children (p=0.032) and higher in women on ART for 2 years or more (p=0.025). Women followed up in a private centre (p<0.001) or a centre with a psychologist (p=0.011) were more susceptible to use contraceptive methods (table 1).

Factors associated with contraceptive use

In multivariate analysis, only five factors remained associated with contraceptive use: education level, marital status, clinical stage, type of health centre and having children. Women with primary education level or those with secondary education level or higher were, respectively, two times more likely (aOR=1.99, 95% CI (1.05 to 3.76)) and four times more likely (aOR=3.95, 95% CI (2.03 to 7.67)) to use a contraceptive method than those who did not have any education, whereas participants living in couple were 55% less likely (aOR=0.45, 95% CI (0.28 to 0.74)) to use a contraceptive method. Women who reported having children and those followed up in a private structure were almost three times more likely (respectively, aOR=2.51, 95% CI (1.41 to 4.5) and aOR=2.54, 95% CI (1.22 to 5.29)) to report current use of contraceptive. Patients at AIDS clinical stage III or IV were 53% less likely (aOR=0.47, 95% CI (0.23 to 0.87)) to use a contraceptive method than those at clinical stage I. In addition this likelihood of using contraceptive is even lower for HIV-infected women at AIDS clinical stage III or IV followed up in private centres (aOR=0.13, 95% CI (0.03 to 0.64)) (table 2).

DIsCussIon

In this study, conducted in two regions in north of Togo, 96.7% of HIV-infected women had heard about contraceptive methods and mainly from health providers. Education level, marital status, AIDS clinical stage, type of health centre and having children had influenced the use of contraceptive methods among HIV-infected women.

A high proportion of knowledge on contraceptive methods was also reported among women aged 15-49 years in the general population in Togo (96.3%) in 2013-2014 [START_REF]et de l'Aménagement du Territoire[END_REF] and among HIV-infected women on ART in Ethiopia (97.0%) in 2013. [START_REF] Abeje | Demand for family planning among HIV positive women on ART: the case of South Gondar and North Wollo Zones Amhara region[END_REF] This showed that seropositive women were as exposed to reproductive health Open Access Open Access information as the rest of the general population, which is a major step towards the use of contraceptive methods. We also found that almost three-quarters (74.7%) of the participants were using a contraceptive method. This proportion was higher than that of 19.9% reported in the general population in 2013 in the Demographic and Health Surveys (DHS) in Togo. [START_REF]et de l'Aménagement du Territoire[END_REF] This was evidenced in prior studies conducted in Sub-Saharan Africa, 12 19 which reported that HIV-infected women seemed to be more likely to use contraceptives than HIV-negative women in public health facilities. This could be explained by the fact that HIV-infected women are frequently in contact with health workers and received more information on reproductive health during their follow-up. HIV-positive women are strongly encouraged to use contraceptive methods to prevent mother-to-child transmission of HIV. Similar results were found in South-East Nigeria (73.1% 10 ) or in Ethiopia (71.0% [START_REF] Asfaw | Contraceptive use and method preference among HIV positive women in Addis Ababa, Ethiopia: a cross sectional survey[END_REF] ), where a high proportion of HIV-infected women used contraceptives. However, lower rates of contraceptive use were reported among HIV-positive mothers in some previous studies in Ethiopia (45.7% [START_REF] Abeje | Demand for family planning among HIV positive women on ART: the case of South Gondar and North Wollo Zones Amhara region[END_REF] ), Uganda (45% [START_REF] Nieves | The influence of partnership on contraceptive use among HIV-infected women accessing antiretroviral therapy in rural Uganda[END_REF] ) or Ghana (42.6% [START_REF] Laryea | Contraceptive use and unmet need for family planning among HIV positive women on antiretroviral therapy in Kumasi, Ghana[END_REF] ). These differences might be due to temporal and regional variabilities, including policy guidelines on reproductive health. In fact cultural characteristics and beliefs, including the perception of the value of a child in the community, may vary through African regions, and this could strongly influence the decision of using a contraceptive method or not. In addition, in Togo, healthcare providers in HIV clinics have recently benefited from a programme of strengthening their skills on family planning. Non-governmental organisations have also been established to promote reproductive health through the distribution of contraceptive methods. This would have encouraged HIV-infected women to use these methods.

As reported in previous studies in Western Africa, 9-11 condom was the most common contraceptive method used by HIV-infected women in this study. Fearing possible pharmacokinetic interactions between hormonal contraceptive and antiretroviral in people living with HIV, 21 22 healthcare providers more often advised HIV-infected women to avoid hormonal contraceptive method in favour of condoms. Condoms, in addition to being accessible and very effective in preventing unwanted pregnancies, contribute to the secondary prevention of HIV, especially in this context where more than four out of ten HIV-infected women did not know the HIV serostatus of their sexual partner. It is therefore important to know the influence of the HIV-infected woman in the decision-making process of condom use or not in a couple.

In this study, the decision to use contraceptive methods in HIV-infected women was influenced by several factors, including education level, marital status, AIDS clinical stage, type of health centre and having children.

Our findings revealed that women who attended school regardless of level (primary, secondary or higher education) were more likely to use contraceptives than those who could not read and write. Women education, which

Open Access

is key to societal development, remains a strong determinant of contraceptive use in Sub-Saharan African. This result was consistent with different studies conducted in Ethiopia 23 24 and in Malawi. [START_REF] Habte | Family planning use among women living with HIV: knowing HIV positive status helps -results from a national survey[END_REF] Women who attended school were able to better analyse reproductive health information, and to understand the mechanisms of action, and the advantages and disadvantages of different contraceptive methods. This could reassure them about the use of these methods and allow them to make a judicious choice.

Our study showed that HIV-infected women who were living with a partner at the time of the survey were less likely to use contraceptive methods. In fact women in couple were more likely to express their desire to have children. This desire is increased by social pressure, which underlines a great importance of having children in the well-being of the couple. In addition, unmarried women would use contraceptive methods more than married women to avoid unwanted pregnancies, which could deteriorate their social image. Unlike our finding, Worke et al [START_REF] Worke | Utilization of contraception among sexually active HIV positive women attending art clinic in University of Gondar Hospital: a hospital based cross-sectional study[END_REF] in Ethiopia reported that married women were more likely to use contraceptives than those who were divorced or separated.

In our study, HIV-infected women followed up in private centres were more likely to use a contraceptive method. Although very few studies compared the quality of healthcare between the public and private sectors, particularly in family planning services delivery, it is generally evidenced that healthcare is better in private health centres. 26 27 In private centres, even though they are non-profit-making facilities, it is reported that care providers are motivated to maximise patients' satisfaction in order to retain them, and this through the provision of good-quality healthcare. However, prior studies 28 29 reported that public health facilities were more likely to offer family planning services. Therefore, it is important to improve the quality of family planning counselling and services delivery in public health facilities in order to encourage patients, including HIV-infected women, to use these services.

Among HIV-infected women, we found that the odds of contraceptive use were lowest at high AIDS clinical stage. Indeed patients at stages III and IV were less likely to use contraceptive methods. At these stages of the disease, patients were likely to suffer from opportunistic infections with certainly a reduction of sexual activities. This could impact on their demand for reproductive health services, including contraceptive use. Moreover, the odds of using contraceptives are even lower when women are followed up in a private centre. This could be explained by the fact that in our setting, HIV-infected patients in the advanced stage of the disease are most often referred for treatment in public hospitals.

HIV-infected women who had children, regardless of number, were more likely to use a contraceptive method. This result is consistent with prior studies conducted in Uganda, [START_REF] Muyindike | Contraceptive use and associated factors among women enrolling into HIV care in southwestern Uganda[END_REF] Malawi 25 and Ethiopia, 23 31 which reported that contraceptive use increased with the number of children. In fact, among women who gave birth at least once, it is very common for them to use contraceptive methods to space or limit births, especially for those who have already reached the number of children they want.

limitations First, the sample may not be representative of the whole country because of cultural disparities across the country. Second, this study took place in health facilities with little chance of including patients who are less regular in the health centres and may have different characteristics. Sexually active women were defined as those engaging in sexual activity in the past 6 months; this definition used in the study could differ from other similar studies. In addition, we lacked information/analysis by contraceptive types, which can impact mother-to-child transmission. Finally, we relied on self-reported reproductive health behaviour through a face-to-face interview, which may introduce information bias. However, these biases would have had very negligible effects on the estimates in this study.

ConClusIon

This study demonstrated a high prevalence of contraceptive use among HIV-infected women followed up in health facilities in Centrale and Kara regions in Togo. Private health facilities seemed to favour the use of contraceptive methods, but not in women at clinical stages III and IV. Despite these results, there is a need for an intensified effort to increase the reproductive health service utilisation particularly in public health facilities in these regions. HIV caregivers must also integrate family planning services in HIV care during follow-up visits.

Table 1

 1 Characteristics of HIV-infected women in the study population (n=461)

	Age <25 years 25-34 years 35-49 years Education level No education Primary Secondary and higher Profession of participants Public sector Private sector Informal sector No profession Marital status In couple Single Residence of patients Urban Rural Region Centrale Kara Religion None	n (%) 40 (8.7) 193 (41.9) 228 (49.5) 65 (14.1) 187 (40.6) 209 (45.3) 34 (7.4) 20 (4.3) 234 (50.8) 173 (37.5) 252 (54.7) 209 (45.3) 287 (62.3) 174 (37.7) 267 (57.9) 194 (42.1) 41 (8.9)	Contraceptive use No, n (%) 8 (20.5) 46 (24.7) 59 (26.7) 27 (42.9) 51 (28.5) 35 (17.2) 7 (20.6) 4 (20.0) 68 (29.7) 34 (20.9) 75 (30.9) 38 (18.7) 71 (25.3) 42 (25.5) 73 (28.6) 40 (20.9) 11 (26.8)	Yes, n (%) 31 (79.5) 140 (75.3) 162 (73.3) 36 (57.1) 128 (71.5) 169 (82.8) 27 (79.4) 16 (80.0) 160 (70.3) 129 (79.1) 168 (69.1) 165 (81.3) 210 (74.7) 123 (74.5) 182 (71.4) 151 (79.1) 30 (73.2)	P values 0.694 <0.001 .192 0.003 0.965 0.065 0.061	BMJ Open: first published as 10.1136/bmjopen-2017-019006 on 24 April 2018. Downloaded from
	Islam Christianism WHO clinical stage Stage I Stage II Stages III and IV Symptom intensity	155 (33.6) 265 (57.5) 252 (55.0) 125 (27.3) 81 (17.6)	48 (31.8) 54 (21.3) 44 (17.9) 36 (30.0) 32 (41.6)	103 (68.2) 200 (78.7) 202 (82.1) 84 (70.0) 45 (58.4)	<0.001 0.018	http://bmjopen.bmj.com/
	None Moderate Severe CD4 cell counts <0.35 × 10 9 /L ≥ 0.35 × 10 9 /L ART Yes No ART scheme (n=415) First line Second line ART duration (=414) <2 years	197 (42.7) 242 (52.5) 22 (4.8) 117 (25.4) 344 (74.6) 415 (90.0) 46 (10.0) 382 (92.0) 33 (8.0) 120 (29.0)	53 (27.6) 51 (21.6) 9 (50.0) 29 (25.9) 84 (25.1) 101 (25.1) 12 (27.3) 96 (26.0) 5 (15.2) 38 (32.8)	139 (72.4) 185 (78.4) 9 (50.0) 83 (74.1) 250 (74.9) 301 (74.9) 32 (72.7) 273 (74.0) 28 (84.8) 78 (67.2)	0.876 0.756 0.168 0.025 Continued	on December 11, 2019 by guest. Protected by copyright.

Table 1

 1 Continuedon December 11, 2019 by guest. Protected by copyright.

	http://bmjopen.bmj.com/

Table 2

 2 Factors associated with contraceptive use among 461 HIV-infected women in the Centrale and Kara regions in Togo

		Logistic regression models
		Univariate	Multivariate
		OR (95% CI)	OR (95% CI)
	Education level	<0.001	<0.001
	No education	1	1
	Primary	1.88 (1.04 to 3.41)	1.99 (1.05 to 3.76)
	Secondary and higher	3.62 (1.95 to 6.72)	3.95 (2.03 to 7.67)
	Profession	0.192
	No profession	1
	Public sector	1.02 (0.41 to 2.53)
	Private sector	1.05 (0.33 to 3.36)
	Informal sector	0.62 (0.39 to 1.00)
	Marital status	0.004	0.001
	Single	1	1
	In couple	0.52 (0.33 to 0.81)	0.45 (0.28 to 0.74)
	Region	0.066
	Centrale	1
	Kara	1.51 (0.97 to 2.36)
	Religion of participants	0.063
	None	1
	Islam	0.79 (0.36 to 1.7)
	Christianism	1.36 (0.64 to 2.89)
	WHO clinical stage	<0.001	0.101
	Stage I	1	1
	Stage II	0.51 (0.31 to 0.85)	0.7 (0.37 to 1.33)
	Stages III and IV	0.31 (0.18 to 0.54)	0.47 (0.24 to 0.94)
	ART scheme	0.175
	First line	0.51 (0.19 to 1.35)
	Second line	1
	Type of health centre	<0.001	0.013
	Public	1	1
	Private	2.46 (1.52 to 3.98)	2.54 (1.22 to 5.29)
	Number of nurses in the	0.001
	centre
	1-10	1
	11-20	0.37 (0.19 to 0.75)
	21 and more	0.42 (0.25 to 0.69)
	Presence of psychologist in	0.011
	the centre
	No	1
	Yes	1.74 (1.13 to 2.69)
	Having children	0.033	0.004
	No	1	1
	Yes	1.72 (1.04 to 2.83)	2.51 (1.41 to 4.5)
	WHO clinical stage*type of		0.041
	health centre †
	Stage I*public centre		1
	Stage II*private centre		0.54 (0.16 to 1.81)
	Stages III and IV*private		0.13 (0.03 to 0.64)
	centre
	ART, antiretroviral therapy.
	*WHO clinical stage.
	†Type of health center.

on December 11, 2019 by guest. Protected by copyright. http://bmjopen.bmj.com/ BMJ Open: first published as 10.1136/bmjopen-2017-019006 on 24 April 2018. Downloaded from

Yaya I, et al. BMJ Open 2018;8:e019006. doi:10.1136/bmjopen-2017-019006

Acknowledgements We would like to thank the health workers involved in data collection for their contribution. We acknowledge all patients who agreed to participate in this study.

ethics approval This study was approved by the National AIDS and STI Programme of Togo (Ref N° 098/2016/MS/DSSP/PNLS-IST).

Provenance and peer review Not commissioned; externally peer reviewed.

Data sharing statement All

Funding This research received no specific grant from any funding agency in the public, commercial or not-for-profit sectors.

data from this study can be accessed and obtained through the corresponding author on request.

Open Access the overall scientific management of the study, for analysis and interpretation, and the preparation of the final manuscript. All the authors read and approved the final manuscript to be submitted for publication.


Thouraya Benmoussa
email: benmoussa_thouraya@yahoo.fr

Oula Amrouni

Laurent Dezileau

Yu Fisjak

Domenico Chiarella

Saadi Abdeljaouad

Gil Mahé

Saâdi Abdeljaouad

Recent Geochemical and Grain Size Distribution of Terrestrial Sediment in Coastal Area from the Watershed of Medjerda River, Gulf of Tunis

Keywords: Coastal, Medjerda river, Extreme floods Turbidities layers, Mining

published or not. The documents may come   L'archive ouverte pluridisciplinaire

Introduction

Rivers are sources of water for social and economic development with the alluvial plain of offering a relatively flat area for urban and agricultural activities. However, human societies have to deal with the fact that the flow of water in river basins is never constant. An often high flow of water during the extreme floods events is one of the more types of natural disasters. Moreover, the sediment leaching from the watershed causes coastal contamination through urban, industrial and agricultural discharges. Rivers are considered as the principal sediment delivery toward the marine environments and are closely depended on the climatic and the human pressure [START_REF] Flemming | Estuarine and coastal geology and geomorphology-A synthesis[END_REF][START_REF] Syvitski | Sediment flux and the anthropocene[END_REF]. The Medjerda River basin in NE-Tunisia is one of the largest basins in the northern zone of Africa. It is located in the northern bay of the Gulf of Tunis between 37°10'N-10°16'E and 37°55'N-10°18'E. The aim of the present study is to characterize the recent geochemical and grain size distribution of the terrestrial sediment in coastal area. out by using the Backman Coulter LS 13 320. A semi-quantitative geochemical analysis was performed by using the X-ray fluorescence (XRF) mobile. To flow the trace terrestrial sediment geochemically, the Zn/Fe; Pb/Fe and Zr/Fe inter-elements ratios have been used. This method provides the advantage to be insensitive to dilution and matrix effects [START_REF] Croudace | ITRAX: description and evaluation of a new multi-function X-ray core scanner[END_REF].

Results

Spatial and Temporal Grain Size

Distribution and Turbidities Layers Identification

Litho Stratigraphic and Sedimentological Description of Turbidity Layers

The results of the grain size distribution of down core sediment are summarized in the Fig. 1. The CEM-1 core sediment shows litho stratigraphic and grain size variability. The down core was roughly divided, from bottom to top, into three major units of granulometric facies: fine sand, silt-clay and medium sand/shell. In the silt-clay facies, 6 coarser turbidities layers are identified from TL-1, TL-2, TL-3, TL-4, TL-5, TL-6, TL-7 and TL-8. These TL layers consist of fine-grained sediments showing multimodal grain size distribution.

Spatial Grain Size Distribution

The Sand/Shell facies of CEM-1(0 and 1 cm depth) is characterized by multimodal grain size distribution. In particular, it is composed of medium sand (200-315 µm) and shells (1200 µm). The riverbanks deposits (MED-1) display medium sand with no occurrence of shell fractions. The sandy beach sediment (MED-2, MED-3 and MED-Nord) is characterized by unimodal grain size distribution with medium sand (250-315 µm). The submarine samples (O10 and O20) collected in the near shore zone display a unimodal grain size distribution with fine sand (100-125 µm).

Geochemical Tracer's Evolution of Sediment Sources

The geochemical evolution of sediment in the different environments is summarized in the Fig.

Discussion

The grain size analysis of surface samples reveals that the coarser deposits present in river consist of heterogeneous medium sand. The sediment of sand beach and the near shore deposits are characterized by unimodal grain size distribution with medium sand. The deltaic deposit is mostly represented of heterogeneous multimodal sediment. However, the turbidity layers are characterized by multimodal grain size distribution. They are formed by three main grain size facies (clay, silt and very fine sand). The geochemical analysis of the surface and core sediments shows that the marine sediments source is represented by Ca and Zr and particularly the basal sand of cores. The Rb, Ti, Zn and Pb constitute the terrestrial sediment source. Silt and clay are generally rich in terrigenous elements. The level peaks of ETM (Pb, Zn) are associated with fine terrestrial sediment (clay and silt). The strongly concentration of Zr is associated with turbidity layers and the surf zone deposits (O20). The Zr is generally associated with aeolian transport deposits [START_REF] Affouri | Extreme flood events reconstruction during the last century in the El Bibane lagoon (Southeast of Tunisia): a multi-proxy approach[END_REF]. Zr is a heavy metal that requires a strong dynamic energy to be mobilized in the coastal area.

The multi proxy results (Fig. 3) reveal that the turbidity layers are characterized by terrestrial sediment source and strongly mining contamination of Zn and Pb. Accordingly, we associate the turbidity layers with high floods events. Based on previous results of [START_REF] Benmoussa | Impact of the Medjerda Sedimentary Fluxes on the Morphodynamic Equilibrium of the Northern Coast of the Gulf of Tunis (Medjerda-Raoued coast)[END_REF] completed by the historical annual rainfall, we associate the TL-1; TL-2; TL-3 and TL-4 to four exceptional floods of Medjerda (1953, 1957, 1969 and 1973) (Fig. 3). These events caused the leaching of old mining discharges into the Medjerda watershed responsible for the coastal sediment contamination of Pb and Zn since 1950.

Conclusion

The terrestrial sediment which feeds the northern coast of the Gulf of Tunis is characterized by the very fine-grained sediment (clay and silt). The geochemical signature of the terrestrial sediment is characterized by highly concentration of Rb, Ti, Zn and Pb. The Medjerda River is the mainly source of silts and clay sediment for the northern coast of the Gulf of Tunis. Even during high frequency events, the sediments are devoid of any coarse fraction. Besides, the mining pollution

Fig. 1

 1 Fig. 1 The litho stratigraphic log description and grain size distribution on down core sediment. Unit-1: Major Sand layer facies; Unit-2: Major silt-clay facies; Unit-3: Major Sand/shell facies. TL-1, TL-2 TL-3 TL-4 TL-5 and TL-6: Turbidity layers formed by heterogeneous facies (Silt, Clay and sand)

Fig. 2

 2 Fig.2The geochemical elements evolution of the Rubidium (Rb), the Titanium (Ti) the calcium (Ca), the Zinc (Zn), the Lead (Pb) and the Zerconium evolution in the sediments samples in the River, sandy beach, deltaic deposits and the nearshore of the Medjerda river coast

Fig. 3

 3 Fig.3The extreme floods identification based on litho stratigraphic log, median grain size (Me); geochemical elements concentration peaks and historical annual rainfall

(i.e. Zn and Pb) is relatively strong in the coastal area especially during the great floods episodes of watershed leaching of Medjerda River.


Floriane Gaucher

Sylvie Bonnassie

Houem Rabah

Pauline Leverrier

Sandrine Pottier

Julien Jardin

Valérie Briard-Bion

Pierre Marchand

Val Erie Briard-Bion

Romain Jeantet

Philippe Blanc

Gw Ena € El Jan

Data from a proteomic analysis highlight different osmoadaptations in two strain of Propionibacterium freudenreichii

Keywords:

a b s t r a c t

The article presents a proteomic data set generated by a comparative analysis of the proteomes of Propionibacterium freudenreichii, comparing the CIRM-BIA 129 and CIRM-BIA 1025 strains. The two strains were cultivated until the beginning of the stationary phase in a chemical defined medium (MMO), and in this medium in the presence of NaCl, with or without glycine betaine. Whole-cell proteins were extracted, trypsinolyzed and analyzed by nano LC-MS/MS, prior to identification and classification by function using the X!Tandem pipeline software and the proteomic data from NCBI.nlm.nigh.gov. Quantification of proteins was then carried out in order to detect change in their expression depending on the culture medium. This article is related to the research article entitled "Benefits and drawbacks of osmotic adjustment in Propionibacterium freudenreichii". The comparative proteomic analysis

Data

This article presents a dataset generated during a proteomic comparative analysis of the cellular proteome of two phenotypical different Propionibacterium freudenreichii strains (CIRM-BIA 129 and CIRM-BIA 1025). The list of all proteins identified is provided in Supplementary Table 1, which presents the protein relative abundances. Fig. 1 shows a heatmap corresponding to abundances, for the two strains and the three culture media (chemically defined medium, the same with salt, the same with salt and glycine betaine). This report presents data from the research article entitled "Benefits and drawbacks of osmotic adjustment in Propionibacterium freudenreichii". The cellular proteomes reveal diversity in the osmoadaptation of P. freudenreichii, among strains.

The proteomic data acquired during this study are presented in Supplementary Table 1 which summarizes all the mean values and all protein with a ratio above 1.5 or below 0.66. For each biological and each technical replicate, a raw data file is presented (https://doi.org/10.15454/VVNQHR).

Fig. 1 summarizes all the observed inductions and repressions. It is composed of a heatmap corresponding to abundances, for the two strains and the three culture media (chemically defined medium, the same with salt, the same with salt and glycine betaine). In press

Value of the Data

This dataset is an important step in understanding the molecular mechanisms responsible for osmoadaptation in P. freudenreichii.

A chemically defined medium was developed to clearly distinguish the effect of 1) osmotic constraint imposed by salt and 2) osmoadaptation in the presence of the universal osmoprotectants glycine betaine in a probiotic bacterium. Surprisingly, molecular mechanisms of osmoadaptation are shown here to vary among strains within the same species. cultivated in MMO (Medium Minus Osmoprotectants). MMO is a synthetic medium which was derivated from CdM (Chemiclal defined Medium) describe previously [START_REF] Gagnaire | Emmental cheese environment enhances Propionibacterium freudenreichii stress tolerance[END_REF], MMO medium has the same composition as CdM but glutamate, glutamine and proline, which are considered as potent sources of osmoprotectants, were removed. P. freudenreichii was grown at 30 C without agitation under microphilic condition. Composition of MMO is detailed in Supplementary Table 1.

Whole cell protein extraction and trypsin digestion

The label free proteomics has have been conducted as Huang et al., 2018 [START_REF] Huang | Growth in hyper-concentrated sweet whey triggers multi stress tolerance and spray drying survival in Lactobacillus casei BL23: from the molecular basis to new perspectives for sustainable probiotic production[END_REF]. At the beginning of stationary phase, P. freudenreichii cells were harvested by centrifugation and washed twice with PBS buffer (NaCl 8g.L À1 , KCl 2g.L À1 KH 2 PO 4 2g.L À1 , Na 2 HPO 4 12H 2 O 35,8g.L À1 ). Cell pellets were then resuspended in lysis solution (lysis solution: 0.5 mL pH 7.5, 157g Tris-HCl, 0.03 g SDS, 0.3 g DTT and 9.5 mL H 2 O), with 1 mM phenylmethylsulfonyl fluoride (PMSF, Sigma-Aldrich, USA). The solution was frozen for 1 hour, then sonicated (2 min 30 HZ), and cells were broken using zirconium beads in the homogenizer (Homog en eisateur Precellys Evolution -Bertin Instruments, France). The resulting SDS extracts were recovered by centrifugation (21,000 Âg; 20 C; 20 min) and then cleaned and quantified using the two-dimensional (2-D) clean-up kit (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) and the 2-D quant kit (GE Healthcare Bio-Sciences AB, Uppsala, Sweden), respectively. Tryptic digestion was performed on 100 mg of whole-cell proteins from each sample during 15 hours at 37 C using Sequencing Grade Modified Trypsin (Promega, Madison, USA) according to the manufacturer's instructions and as described previously [START_REF] Huang | Growth in hyper-concentrated sweet whey triggers multi stress tolerance and spray drying survival in Lactobacillus casei BL23: from the molecular basis to new perspectives for sustainable probiotic production[END_REF]. Spectrophotometric-grade trifluoroacetic acid (TFA) (Sigma-Aldrich, USA) was added in order to stop the digestion.

Nano LC-MS/MS

Experiments were performed as previously described [START_REF] Huang | Growth in hyper-concentrated sweet whey triggers multi stress tolerance and spray drying survival in Lactobacillus casei BL23: from the molecular basis to new perspectives for sustainable probiotic production[END_REF]. Experiments were performed using a nano RSLC Dionex U3000 system fitted to a Q-Exactive mass spectrometer (Thermo Scientific, San Jose, USA) equipped with a nano-electrospray ion source. A preliminary sample concentration step was performed on a C18 pepMap100 reverse phase column (C18 column, 300-mm inner diameter (i.d.), 5 mm length, 5 mm particle size, 100 Å pore size; Dionex, Amsterdam, The Netherlands). Peptides separation was performed on a reversed-phase column (PepMap 100 C18, 75 mm i.d., 250 mm length, 3 mm particle size, 100 Å pore size; Dionex, Amsterdam, The Netherlands) with a column temperature of 35 C, using solvent A (2% (v/v) acetonitrile (Honeywell, USA), 0.08% (v/v) formic acid and 0.01% (v/v) TFA in deionized water) and solvent B (95% (v/v) acetonitrile, 0.08% (v/v) formic acid and 0.01% (v/v) TFA in deionized water). Peptides were separated using a gradient of 5e35% solvent B over 80 min followed by 35e85% solvent B over 5 min at a flow rate of 0.3 mL/min. Eluted peptides were directly electro sprayed into the mass spectrometer operating in positive ion mode with a voltage of 2kV. Spectra were recorded in full MS mode and selected in a mass range 250e2000 m/z for MS spectra with a resolution of 70,000 at m/z 200. For each scan, the ten most intense ions were selected for fragmentation. MS/MS spectra were recorded with a resolution of 17,500 at m/z 200 and the parent ion was subsequently excluded from MS/MS fragmentation for 20 s. The instrument was externally calibrated according to the supplier's instructions.

Proteins identification

Proteins identification was performed as previously described [START_REF] Huang | Growth in hyper-concentrated sweet whey triggers multi stress tolerance and spray drying survival in Lactobacillus casei BL23: from the molecular basis to new perspectives for sustainable probiotic production[END_REF]. Peptides were identified from the MS/MS spectra using the X!Tandem pipeline software (Langella et al., 2017). The search was performed against a database composed of proteomes of strains P. freudenreichii CIRM-BIA 129 and CIRM-BIA 1025 (downloaded from NCBI.nlm.nih.gov on the August 23, 2018). Database search parameters were specified as follow: trypsin cleavage was used and the peptide mass tolerance was set to 10 ppm for MS and 0.05 Da for MS/MS. Oxidation of methionine and phosphorylation of threonine, serine and tryptophan were selected as a variable modification. For each peptide identified, a minimum score corresponding to an e-value below 0.05 was considered as a prerequisite for peptide validation. Please cite this article as: F. Gaucher et al., Data from a proteomic analysis highlight different osmoadaptations in two strain of Propionibacterium freudenreichii, Data in brief, https://doi.org/10.1016/ j.dib.2019.104932

Protein quantification

Protein quantification was performed as previously described [START_REF] Huang | Growth in hyper-concentrated sweet whey triggers multi stress tolerance and spray drying survival in Lactobacillus casei BL23: from the molecular basis to new perspectives for sustainable probiotic production[END_REF]. Every peptide identified by tandem mass spectrometry was quantified using the free MassChroQ software (Valot et al., 2011) before data treatment and statistical analysis within the R software (R 3.2.2, Project for statistical computing). A specific R package called 'MassChroqR' was used to automatically filter dubious peptides for which standard deviation of retention time was superior to 30 s and to regroup peptide quantification data into proteins. Two different and complementary methods of analysis were used, based on peak counting or XIC (eXtracted Ion Current). For peak counting analysis, variance analysis was performed to detect significant abundance variation between culture conditions. Proteins with an adjusted p-value <0.05 were considered significantly different. For XIC based quantification, normalization was performed to take into account possible global quantitative variations between LC-MS runs. Peptides shared between different proteins were automatically excluded from the data set as well as peptides present in less than 95% of samples. Missing data were then imputed from a linear regression based on other peptide intensities for the same protein [START_REF] Blein-Nicolas | A systems approach to elucidate heterosis of protein abundances in yeast[END_REF]. Protein abundance was computed as the sum of peptide areas requiring a minimum of 2 peptides per protein. For every missing protein abundance, missing value were replaced by the minimum abundance obtained for this protein in the whole experiment. Analysis of variance was used to determine proteins with significantly different abundance between our culture conditions.

Uncited reference [START_REF] Gaucher | Benefits and drawbacks of osmotic adjustment in Propionibacterium freudenreichii[END_REF] Q 5

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F 4 F

 4 Fig. 1.

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Acknowledgments

F. Gaucher is the recipient of a joint doctoral fellowship from Bioprox and from ANRT (Association Nationale de la Recherche et de la Technologie).

Conflict of interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Appendix A. Supplementary data

Supplementary data to this article can be found online at https://doi.org/10.1016/j.dib.2019.104932.