Valorization of Waste Dates in South Algeria: Biofuel Production

In Algeria, the conditioning units of dates, generate significant quantities of waste arising from sorting deviations. This biomass, until then considered as a waste with high impact on the environment can be transformed into high value added product. It is possible to develop common dates of low commercial value, and put on the local and international market a new generation of products with high added values such as bio ethanol. Besides its use in chemical synthesis, bio ethanol can be blended with gasoline to produce a clean fuel while improving the octane.

Effect of PGPB Inoculation, Addition of Biochar, and Mineral N Fertilization on Mycorrhizal Colonization

Strong anthropogenic impact has uncontrolled consequences on the nature of the soil. Hence, up-to-date sustainable methods of soil state improvement are essential. Investigators provide the evidence that biochar can positively effects physical, chemical, and biological soil properties and the abundance of mycorrhizal fungi which are in the focus of this study. The main aim of the present investigation is to demonstrate the effect of two types of plant growth promoting bacteria (PGPB) inoculums along with the beech wood biochar and mineral N additives on mycorrhizal colonization. Experiment has been set up in laboratory conditions with containers filled with arable soil from the protection zone of the main water source “Brezova nad Svitavou”. Lactuca sativa (lettuce) has been selected as a model plant. Based on the obtained data, it can be concluded that mycorrhizal colonization increased as the result of combined influence of biochar and PGPB inoculums amendment. In addition, correlation analyses showed that the numbers of main groups of cultivated bacteria were dependent on the degree of mycorrhizal colonization.

Optimization of NaOH Thermo-Chemical Pretreatment to Enhance Solubilisation of Organic Food Waste by Response Surface Methodology

This study investigates the influence of low temperature thermo-chemical pretreatment of organic food waste on performance of COD solubilisation. Both temperature and alkaline agent were reported to have effect on solubilizing any possible biomass including organic food waste. The three independent variables considered in this pretreatment were temperature (50-90oC), pretreatment time (30-120 minutes) and alkaline concentration, sodium hydroxide, NaOH (0.7-15 g/L). The maximal condition obtained were 90oC, 15 g/L NaOH for 2 hours. Solubilisation has potential in enhancing methane production by providing high amount of soluble components at early stage during anaerobic digestion.

Supramolecular Cocrystal of 2-Amino-4-Chloro-6- Methylpyrimidine with 4-Methylbenzoic Acid: Synthesis, Structural Determinations and Quantum Chemical Investigations

The 1:1 cocrystal of 2-amino-4-chloro-6- methylpyrimidine (2A4C6MP) with 4-methylbenzoic acid (4MBA) (I) has been prepared by slow evaporation method in methanol, which was crystallized in monoclinic C2/c space group, Z = 8, and a = 28.431 (2) Å, b = 7.3098 (5) Å, c = 14.2622 (10) Å and β = 109.618 (3)°. The presence of unionized –COOH functional group in cocrystal I was identified both by spectral methods (1H and 13C NMR, FTIR) and X-ray diffraction structural analysis. The 2A4C6MP molecule interact with the carboxylic group of the respective 4MBA molecule through N—H⋯O and O—H⋯N hydrogen bonds, forming a cyclic hydrogen–bonded motif R2 2(8). The crystal structure was stabilized by Npyrimidine—H⋯O=C and C=O—H⋯Npyrimidine types hydrogen bonding interactions. Theoretical investigations have been computed by HF and density function (B3LYP) method with 6–311+G (d,p)basis set. The vibrational frequencies together with 1H and 13C NMR chemical shifts have been calculated on the fully optimized geometry of cocrystal I. Theoretical calculations are in good agreement with the experimental results. Solvent–free formation of this cocrystal I is confirmed by powder X-ray diffraction analysis.

Comparison of the Effects of Continuous Flow Microwave Pre-treatment with Different Intensities on the Anaerobic Digestion of Sewage Sludge for Sustainable Energy Recovery from Sewage Treatment Plant

Anaerobic digestion is a well-known technique for sustainable energy recovery from sewage sludge. However, sewage sludge digestion is restricted due to certain factors. Pre-treatment methods have been established in various publications as a promising technique to improve the digestibility of the sewage sludge and to enhance the biogas generated which can be used for energy recovery. In this study, continuous flow microwave (MW) pre-treatment with different intensities were compared by using 5 L semi-continuous digesters at a hydraulic retention time of 27 days. We focused on the effects of MW at different intensities on the sludge solubilization, sludge digestibility, and biogas production of the untreated and MW pre-treated sludge. The MW pre-treatment demonstrated an increase in the ratio of soluble chemical oxygen demand to total chemical oxygen demand (sCOD/tCOD) and volatile fatty acid (VFA) concentration. Besides that, the total volatile solid (TVS) removal efficiency and tCOD removal efficiency also increased during the digestion of the MW pre-treated sewage sludge compared to the untreated sewage sludge. Furthermore, the biogas yield also subsequently increases due to the pre-treatment effect. A higher MW power level and irradiation time generally enhanced the biogas generation which has potential for sustainable energy recovery from sewage treatment plant. However, the net energy balance tabulation shows that the MW pre-treatment leads to negative net energy production.

In vitro Susceptibility of Madurella mycetomatis to the Extracts of Anogeissus leiocarpus Leaves

Anogeissus leiocarpus (Combretaceae) is well known for its medicinal uses in African traditional medicine, for treating many human diseases mainly skin diseases and infections. Mycetoma disease is a fungal and/ or bacterial skininfection, mainly cause by Madurella mycetomatis fungus. This study was carried out in vitro to investigate the antifungal activity of Anogeissus leiocarpus leaf extracts against the isolated pathogenic Madurella mycetomatis, by using the NCCLS modified method compared to Ketoconazole standard drug, and MTT assay. The bioactive fraction was subjected to chemical analysis implementing different chromatographic analytical methods (TLC, HPLC, and LC-MS/MS). The results showed significance antifungal activity of A. leiocarpus leaf extracts against the isolated pathogenic M. mycetomatis, compared to negative and positive controls. The chloroform fraction showed the highest antifungal activity. The chromatographic analysis of the chloroform fraction with the highest activity showed the presence of important bioactive compounds such as ellagic and flavellagic acids derivatives, flavonoids and stilbenoid, which are well known for their antifungal activity.

Chemical Characterization and Prebiotic Effect of Water-Soluble Polysaccharides from Zizyphus lotus Leaves

In order to investigate the prebiotic potential of oligosaccharides prepared by chemical hydrolysis of water-soluble polysaccharides (WSP) from Zizyphus lotus leaves, the effect of oligosaccharides on bacterial growth was studied. The chemical composition of WSP was evaluated by colorimetric assays revealed the average values: 7.05±0.73% proteins and 86.21±0.74% carbohydrates, among them 64.81±0.42% is neutral sugar and the rest 16.25±1.62% is uronic acids. The characterization of monosaccharides was determined by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was found to be composed of galactose (23.95%), glucose (21.30%), rhamnose (20.28%), arabinose (9.55%), and glucuronic acid (22.95%). The effects of oligosaccharides on the growth of lactic acid bacteria were compared with those of fructooligosaccharide (RP95). The oligosaccharides concentration was 1g/L of Man, Rogosa, Sharpe broth. Bacterial growth was assessed during 2, 4.5, 6.5, 9, 12, 16 and 24 h by measuring the optical density of the cultures at 600 nm (OD600) and pH values. During fermentation, pH in broth cultures decreased from 6.7 to 5.87±0.15. The enumeration of lactic acid bacteria indicated that oligosaccharides led to a significant increase in bacteria (P≤0.05) compared to the control. The fermentative metabolism appeared to be faster on RP95 than on oligosaccharides from Zizyphus lotus leaves. Both RP95 and oligosaccharides showed clear prebiotic effects, but had differences in fermentation kinetics because of to the different degree of polymerization. This study shows the prebiotic effectiveness of oligosaccharides, and provides proof for the selection of leaves of Zizyphus lotus for use as functional food ingredients.

A DNA-Based Nanobiosensor for the Rapid Detection of the Dengue Virus in Mosquito

This paper describes the development of a DNA-based nanobiosensor to detect the dengue virus in mosquito using electrically active magnetic (EAM) nanoparticles as concentrator and electrochemical transducer. The biosensor detection encompasses two sets of oligonucleotide probes that are specific to the dengue virus: the detector probe labeled with the EAM nanoparticles and the biotinylated capture probe. The DNA targets are double hybridized to the detector and the capture probes and concentrated from nonspecific DNA fragments by applying a magnetic field. Subsequently, the DNA sandwiched targets (EAM-detector probe– DNA target–capture probe-biotin) are captured on streptavidin modified screen printed carbon electrodes through the biotinylated capture probes. Detection is achieved electrochemically by measuring the oxidation–reduction signal of the EAM nanoparticles. Results indicate that the biosensor is able to detect the redox signal of the EAM nanoparticles at dengue DNA concentrations as low as 10 ng/μl.

Poly[3-(3,4-dihydroxyphenyl)Glyceric Acid] with Potential Therapeutic Effect

According to IR, 13C and 1H NMR, APT, 1D NOE, 2D heteronuclear 1H/13C HSQC and 2D DOSY experiments the main chemical constituent of high-molecular preparations from Symphytum asperum, S. caucasicum, S. officinale and Anchusa italica (Boraginaceae) was found to be caffeic acid-derived polyether, namely poly[3-(3,4-dihydroxyphenyl)glyceric acid] (PDPGA) or poly[oxy-1-carboxy-2-(3,4-dihydroxyphenyl)ethylene]. Most carboxylic groups of this polymer of A. italica are methylated.

Noninvasive Disease Diagnosis through Breath Analysis Using DNA-Functionalized SWNT Sensor Array

Noninvasive diagnostics of diseases via breath analysis has attracted considerable scientific and clinical interest for many years and become more and more promising with the rapid advancements in nanotechnology and biotechnology. The volatile organic compounds (VOCs) in exhaled breath, which are mainly blood borne, particularly provide highly valuable information about individuals’ physiological and pathophysiological conditions. Additionally, breath analysis is noninvasive, real-time, painless, and agreeable to patients. We have developed a wireless sensor array based on single-stranded DNA (ssDNA)-functionalized single-walled carbon nanotubes (SWNT) for the detection of a number of physiological indicators in breath. Seven DNA sequences were used to functionalize SWNT sensors to detect trace amount of methanol, benzene, dimethyl sulfide, hydrogen sulfide, acetone, and ethanol, which are indicators of heavy smoking, excessive drinking, and diseases such as lung cancer, breast cancer, and diabetes. Our test results indicated that DNA functionalized SWNT sensors exhibit great selectivity, sensitivity, and repeatability; and different molecules can be distinguished through pattern recognition enabled by this sensor array. Furthermore, the experimental sensing results are consistent with the Molecular Dynamics simulated ssDNAmolecular target interaction rankings. Thus, the DNA-SWNT sensor array has great potential to be applied in chemical or biomolecular detection for the noninvasive diagnostics of diseases and personal health monitoring.

Thermal Method for Testing Small Chemisorbents Samples on the Base of Potassium Superoxide

The increase of technogenic and natural accidents, accompanied by air pollution, for example, by combustion products, leads to the necessity of respiratory protection. This work is devoted to the development of a calorimetric method and a device which allows investigating quickly the kinetics of carbon dioxide sorption by chemisorbents on the base of potassium superoxide in order to assess the protective properties of respiratory protective closed circuit apparatus. The features of the traditional approach for determining the sorption properties in a thin layer of chemisorbent are described, as well as methods and devices, which can be used for the sorption kinetics study. The authors developed an approach (as opposed to the traditional approach) based on the power measurement of internal heat sources in the chemisorbent layer. The emergence of the heat sources is a result of exothermic reaction of carbon dioxide sorption. This approach eliminates the necessity of chemical analysis of samples and can significantly reduce the time and material expenses during chemisorbents testing. Error of determining the volume fraction of adsorbed carbon dioxide by the developed method does not exceed 12%. Taking into account the efficiency of the method, we consider that it is a good alternative to traditional methods of chemical analysis under the assessment of the protection sorbents quality.

Application of Micro-Tunneling Technique to Rectify Tilted Structures Constructed on Cohesive Soil

Foundation differential settlement and supported structure tilting are an occasionally occurred engineering problem. This may be caused by overloading, changes in ground soil properties or unsupported nearby excavations. Engineering thinking points directly toward the logic solution for such problem by uplifting the settled side. This can be achieved with deep foundation elements such as micro-piles and macro-piles™, jacked piers, and helical piers, jet grouted mortar columns, compaction grout columns, cement grouting or with chemical grouting, or traditional pit underpinning with concrete and mortar. Although, some of these techniques offer economic, fast and low noise solutions, many of them are quite the contrary. For tilted structures, with the limited inclination, it may be much easier to cause a balancing settlement on the less-settlement side which shall be done carefully in a proper rate. This principal has been applied in Leaning Tower of Pisa stabilization with soil extraction from the ground surface. In this research, the authors attempt to introduce a new solution with a different point of view. So, the micro-tunneling technique is presented in here as an intended ground deformation cause. In general, micro-tunneling is expected to induce limited ground deformations. Thus, the researchers propose to apply the technique to form small size ground unsupported holes to produce the target deformations. This shall be done in four phases: 1. Application of one or more micro-tunnels, regarding the existing differential settlement value, under the raised side of the tilted structure. 2. For each individual tunnel, the lining shall be pulled out from both sides (from jacking and receiving shafts) in the slow rate. 3. If required, according to calculations and site records, an additional surface load can be applied on the raised foundation side. 4. Finally, a strengthening soil grouting shall be applied for stabilization after adjustment. A finite element based numerical model is presented to simulate the proposed construction phases for different tunneling positions and tunnels group. For each case, the surface settlements are calculated and induced plasticity points are checked. These results show the impact of the suggested procedure on the tilted structure and its feasibility. Comparing results also show the importance of the position selection and tunnels group gradual effect. Thus, a new engineering solution is presented to one of the structural and geotechnical engineering challenges.

Comparative Correlation Investigation of Polynuclear Aromatic Hydrocarbons (PAHs) in Soils of Different Land Use: Sources Evaluation Perspective

Polycyclic Aromatic Hydrocarbons (PAHs) are formed mainly because of incomplete combustion of organic materials during industrial, domestic activities or natural occurrence. Their toxicity and contamination of terrestrial and aquatic ecosystem have been established. However, with limited validity index, previous research has focused on PAHs isomer pair ratios of variable physicochemical properties in source identification. The objective of this investigation was to determine the empirical validity of Pearson Correlation Coefficient (PCC) and Cluster Analysis (CA) in PAHs source identification along soil samples of different land uses. Therefore, 16 PAHs grouped, as Endocrine Disruption Substances (EDSs) were determined in 10 sample stations in top and sub soils seasonally. PAHs was determined the use of Varian 300 gas chromatograph interfaced with flame ionization detector. Instruments and reagents used are of standard and chromatographic grades respectively. PCC and CA results showed that the classification of PAHs along pyrolitic and petrogenic organics used in source signature is about the predominance PAHs in environmental matrix. Therefore, the distribution of PAHs in the studied stations revealed the presence of trace quantities of the vast majority of the sixteen PAHs, which may ultimately inhabit the actual source signature authentication. Therefore, factors to be considered when evaluating possible sources of PAHs could be; type and extent of bacterial metabolism, transformation products/substrates, and environmental factors such as salinity, pH, oxygen concentration, nutrients, light intensity, temperature, co-substrates, and environmental medium are hereby recommended as factors to be considered when evaluating possible sources of PAHs.

Phytochemical Screening, Antioxidant Activity and Lipid Profile Effects of Citrus reticulata Fruit Peel, Zingiber officinale Rhizome and Sesamum indicum Seed Extracts

Many herbal medicinal products are considered potential anti-hypercholesterolemic agents with encouraging safety profiles, however only a limited amount of clinical research exists to support their efficacy. The present study was designed to compare the antihypercholesterolemic and antioxidant activities of the crude ethanolic extracts of Citrus reticulata fruit peel, Zingiber officinale rhizome and Sesamum indicum seeds. Forty-five rats were used throughout the experiment which are extended for four weeks. These were divided into nine groups, five rats per each group as follows; group 1 was the normal control group (rats only fed standard normal rat diet), group 2 was the hypercholesterolemic control group (rats fed only hypercholesterolemic diet which contained 1% cholesterol plus 10% saturated animal fat added to the normal rat diet), groups 3 and 4 were fed hypercholesterolemic diet in addition to Citrus reticulata ethanolic extract at doses of (250mg/kg (group 3) and 500mg/kg (group 4)) administered daily via oral route, groups 5 and 6 were given hypercholesterolemic diet in addition to Zingiber officinale ethanolic extract at doses of (250mg/kg (group 5) and 500mg/kg (group 6)) daily through oral route, groups 7 and 8 fed on hypercholesterolemic diet in addition to Sesamum indicum ethanolic extract at doses of (250mg/kg (group 7) and 500mg/kg (group 8)) daily orally; and group 9 rats were given hypercholesterolemic diet in addition to atorvastatin (0.18mg/kg) daily via oral route as a standard reference antihypercholesterolemic drug. Blood samples from all groups were drawn from the retro-orbital venous plexus four weeks following treatment after overnight fasting and the lipid profile (total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C) and triglyceride levels) were measured and the risk ratio (TC/HDL-C) was assessed. The antioxidant activity of the three plants extracts was determined using DPPH free-radical antioxidant assay. Results of in vivo and in vitro antihypercholesterolemic and antioxidant assay respectively, revealed that the three extracts possess comparable antioxidant and antihypercholesterolemic activities.

A Structural and Magnetic Investigation of the Inversion Degree in Spinel NiFe2O4, ZnFe2O4 and Ni0.5Zn0.5Fe2O4 Ferrites Prepared by Soft Mechanochemical Synthesis

NiFe2O4 (nickel ferrite), ZnFe2O4 (zinc ferrite) and Ni0.5Zn0.5Fe2O4 (nickel-zinc ferrite) were prepared by mechanochemical route in a planetary ball mill starting from mixture of the appropriate quantities of the Ni(OH)2/Fe(OH)3, Zn(OH)2/Fe(OH)3 and Ni(OH)2/Zn(OH)2/Fe(OH)3 hydroxide powders. In order to monitor the progress of chemical reaction and confirm phase formation, powder samples obtained after 25 h, 18 h and 10 h of milling were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), IR, Raman and Mössbauer spectroscopy. It is shown that the soft mechanochemical method, i.e. mechanochemical activation of hydroxides, produces high quality single phase ferrite samples in much more efficient way. From the IR spectroscopy of single phase samples it is obvious that energy of modes depends on the ratio of cations. It is obvious that all samples have more than 5 Raman active modes predicted by group theory in the normal spinel structure. Deconvolution of measured spectra allows one to conclude that all complex bands in the spectra are made of individual peaks with the intensities that vary from spectrum to spectrum. The deconvolution of Raman spectra allows to separate contributions of different cations to a particular type of vibration and to estimate the degree of inversion.

Evaluation of Water Quality for the Kurtbogazi Dam Outlet and the Streams Feeding the Dam in Ankara, Turkey

Kurtbogazi Dam has gained special meaning for Ankara, Turkey for the last decade due to the rapid depletion of nearby resources of drinking water. In this study, the results of the analyses of Kurtbogazi Dam outlet water and the rivers flowing into the Kurtbogazi Dam were discussed for the period of last five years between 2008 and 2012. Some physical and chemical properties (pH, temperature, biochemical oxygen demand (BOD5), nitrate, phosphate and chlorine) of these water resources were evaluated. They were classified according to the Council Directive (75/440/EEC). Moreover, the properties of these surface waters were assessed to determine the quality of water for drinking and irrigation purposes using Piper, US Salinity Laboratory and Wilcox diagrams. The results showed that all the water resources are acceptable level as surface water except for Pazar Stream in terms of ortho-phosphate and BOD5 concentration for 2008.

Estimated Human Absorbed Dose of 111In-BPAMD as a New Bone-Seeking SPECT-Imaging Agent

An early diagnosis of bone metastasis is very important for making a right decision on a subsequent therapy. One of the most important steps to be taken initially, for developing a new radiopharmaceutical is the measurement of organ radiation exposure dose. In this study, the dosimetric studies of a novel agent for SPECT-imaging of the bone metastasis, 111In-(4- {[(bis(phosphonomethyl))carbamoyl]methyl}7,10bis(carboxymethyl) -1,4,7,10-tetraazacyclododec-1-yl) acetic acid (111In-BPAMD) complex, have been carried out to estimate the dose in human organs based on the data derived from mice. The radiolabeled complex was prepared with high radiochemical purity in the optimal conditions. Biodistribution studies of the complex was investigated in the male Syrian mice at the selected times after injection (2, 4, 24 and 48 h). The human absorbed dose estimation of the complex was made based on data derived from the mice by the radiation absorbed dose assessment resource (RADAR) method. 111In-BPAMD complex was prepared with high radiochemical purity >95% (ITLC) and specific activities of 2.85 TBq/mmol. Total body effective absorbed dose for 111In-BPAMD was 0.205 mSv/MBq. This value is comparable to the other 111In clinically used complexes. The results show that the dose with respect to the critical organs is satisfactory within the acceptable range for diagnostic nuclear medicine procedures. Generally, 111In-BPAMD has interesting characteristics and it can be considered as a viable agent for SPECT-imaging of the bone metastasis in the near future.

Up Scaling of Highly Transparent Quasi-Solid State, Dye-Sensitized Solar Devices Composed of Nanocomposite Materials

At the present work, highly transparent strip type quasi-solid state dye-sensitized solar cells (DSSCs) were fabricated through inkjet printing using nanocomposite TiO2 inks as raw materials and tested under outdoor illumination conditions. The cells, which can be considered as the structural units of large area modules, were fully characterized electrically and electrochemically and after the evaluation of the received results a large area DSSC module was manufactured. The module design was a sandwich Z-interconnection where the working electrode is deposited on one conductive glass and the counter electrode on a second glass. Silver current collective fingers were printed on the conductive glasses to make the internal electrical connections and the adjacent cells were connected in series and finally insulated using a UV curing resin to protect them from the corrosive (I-/I3-) redox couple of the electrolyte. Finally, outdoor tests were carried out to the fabricated dye-sensitized solar module and its performance data were collected and assessed.

Preliminary Dosimetric Evaluation of a New Therapeutic 177Lu Complex for Human Based on Biodistribution Data in Rats

Abstract—[Tris (1,10-phenanthroline) lanthanum(III)] trithiocyanate is a new compound that has shown high ability for stopping the synthesis of DNA and also acting as a photosensitizer. Nowadays, the radiation dose assessment resource (RADAR) method is known as the most common method for absorbed dose calculation. 177Lu was produced by (n, gamma) reaction in a research reactor. 177Lu-PL3 was prepared in the optimized condition. The radiochemical yield was checked by ITLC method. The biodistribution of the complex was investigated by intravenously injection to wild-type rats via their tail veins. In this study, the absorbed dose of 177Lu-PL3 to human organs was estimated by RADAR method. 177Lu was prepared with a specific activity of 2.6-3 GBq.mg-1 and radionuclide purity of 99.98 %. Final preparation of the radiolabelled complex showed high radiochemical purity of > 99%. The results show that liver and spleen have received the highest absorbed dose of 1.051 and 0.441 mSv/MBq, respectively. The absorbed dose values for these two dose-limiting tissues suggest more biological studies special in tumor-bearing animals.

Human Absorbed Dose Estimation of a New IN-111 Imaging Agent Based on Rat Data

The measurement of organ radiation exposure dose is one of the most important steps to be taken initially, for developing a new radiopharmaceutical. In this study, the dosimetric studies of a novel agent for SPECT-imaging of the bone metastasis, 111In- 1,4,7,10-tetraazacyclododecane-1,4,7,10 tetraethylene phosphonic acid (111In-DOTMP) complex, have been carried out to estimate the dose in human organs based on the data derived from rats. The radiolabeled complex was prepared with high radiochemical purity in the optimal conditions. Biodistribution studies of the complex was investigated in the male Syrian rats at selected times after injection (2, 4, 24 and 48 h). The human absorbed dose estimation of the complex was made based on data derived from the rats by the radiation absorbed dose assessment resource (RADAR) method. 111In-DOTMP complex was prepared with high radiochemical purity of >99% (ITLC). Total body effective absorbed dose for 111In- DOTMP was 0.061 mSv/MBq. This value is comparable to the other 111In clinically used complexes. The results show that the dose with respect to the critical organs is satisfactory within the acceptable range for diagnostic nuclear medicine procedures. Generally, 111In- DOTMP has interesting characteristics and can be considered as a viable agent for SPECT-imaging of the bone metastasis in the near future.