Egg yolk (20%; v/v) was added to the each extender and mixed by placing the tube to an orbital shaker for 10 min and centrifuged at 15,000g for 60 min, and the supernatant was filtered through a 0.45 μm membrane filter. Egg yolk phospholipids were then solubilized by adding 0.75% (v/v) Equex-Paste (Minitüb, Tiefenbach, Germany) to the extender. Sperm samples (100 μl) from SD or F344 were transferred into 1.5 ml centrifuge tubes containing 400 μl of each freezing extender and gently mixed by inverting the tube. After dilution, motility analysis was performed

using a phase Akt inhibitor contrast microscopy equipped with 20× objective. The sperm samples were then equilibrated at 4 °C for 45 min. After equilibration in the extenders, 150 μl sperm sample from each extender was placed onto a shallow quartz dish (14 mm inner diameter and 2.56 mm deep) and covered with a round coverslip and then inserted into Linkam cryostage (TMS-94) that was mounted on a Nikon microscope.

The AZD2281 order samples were then cooled by using various cooling rates (10, 40, 70 and 100 °C/min) to final temperature of −150 °C. For thawing, the quartz dish containing the sperm samples was rapidly removed from the Linkam cryostage and placed on a 37 °C slide warmer in order to have direct contact with the warm surface to achieve about 1000 °C/min warming rate. After warming, motility analysis was performed and the samples were transferred into 1.5 mL Eppendorf tubes containing 150 μL TL-HEPES base solution. All samples were underwent mitochondrial, acrosome and membrane integrity assessment. SYBR-14/Propidium iodide (Live/Dead sperm viability kit, catalog no: L-7011, Molecular Probes, Eugene, OR, USA)

and Alexa Fluor-488-PNA (catalog no: L-21,409, Molecular Probes, Eugene, OR, USA) conjugate were used to determine rat sperm plasma membrane and acrosome integrity, respectively. For plasma membrane integrity, 200 μl TL-HEPES Fossariinae solution was gently added to the tube containing 100 μl thawed sperm (1–2 × 106 spermatozoa/ml). Diluted sperm samples were incubated with 5 μL PI (0.5 μM final concentration) and 10 μL (0.4 μM final concentration) SYBR-14 at 37 °C for 10 min. After staining, 10 μl of sperm sample was placed on a microscope slide, covered with a coverslip and observed under the epifluorescence microscope (Nikon Eclipse 600 using a dual fluorescence filter). The images of stained sperm samples were classified into two groups: sperm head displaying green fluorescence was considered to be membrane intact, whereas sperm displaying red fluorescence in the head was considered to be damaged membrane. 100 sperm per sample were counted as described previously [52]. To evaluate sperm acrosomal integrity, thawed sperm samples were washed to remove freezing extender.

For inoculating the fungus expressing DsRed, the maize roots 1 cm in length were immersed in a suspension of spores (105 conidia mL− 1) for 5 min before transfer

to 1 mL of BNM medium on a slide. The growth and colonization of F. verticillioides were subjected to epifluorescent microscopy. Following infection with F. verticillioides, the dyes of neutral red (0.01%, W/V) and Evans blue (0.2%, W/V) (Sigma, St. Louis, MO, USA) were used to stain the cross and longitudinal sections of the roots for 5 min each, rinsed with water, and then observed under a microscope. GSI-IX molecular weight Dead cells were stained blue with Evans blue, whereas living cells were stained red with neutral red. Certain characteristic indicators, e.g., accumulation of peroxide, can be detected when PCD occurs. To investigate whether infection of F. verticillioides induced PCD in maize leaves, peroxide

staining using 3,3-diaminobenzidine (DAB) as the substrate was performed to detect the accumulation of H2O2 following infection of F. verticillioides as described previously [33]. At the two-leaf-stage, the leaves of maize plants inoculated with the F. verticillioides strain expressing DsRed were excised and incubated in a 1 mg mL− 1 solution of DAB (pH 3.8) for 2 h under light at 25 °C and then boiled in ethanol (96%) for 10 min. After cooling, the leaves were extracted with fresh ethanol at room temperature. The degree of dark brown polymerization indicated the amount of H2O2 accumulated in the treated leaves. MK-2206 ic50 Selective Fusarium Idelalisib agar (SFA) [29] and [30] medium was used to analyze the colonization of F. verticillioides on/in maize roots. DsRed-labeled fungus-infected and mock-inoculated roots and basal stems of maize were sampled at different times after the inoculation from two replicated greenhouse trials to determine the numbers of colony forming units (CFU) as previously described [31]. A randomized complete block design with four replicates consisting of two plants each was used to arrange the inoculated maize plants. The roots

were removed from the vermiculite, washed thoroughly with tap water, and surface sterilized for 3 min in 0.5% (V/V) NaOCl solution. After rinsing with sterile deionized water several times, roots were wiped with sterile filter paper. Roots and basal stems from the two plants in each replicate were weighted, ground, and mixed into 10 ml of sterile deionized water with a Fast-Prep-24 Instrument (MP Biomedicals, Solon, OH, USA) at high speed for 1 min. Homogenized suspensions of root and basal stem samples were filtered through four layers of cheesecloth to remove plant debris and diluted 20-fold with sterile deionized water. The diluted samples were separately spread with a sterile glass rod onto the SFA plates. Each inoculated sample consisted of five replicated plates with 50 μL of diluted tissue suspension.

, 2010).

Previous studies have observed Z-VAD-FMK datasheet robust vATL activations for semantic tasks using this technique (Binney et al., 2010 and Visser and Lambon Ralph, 2011). Images were acquired on a 3T Philips Achieva scanner using an 8 element SENSE head coil with a sense factor of 2.5. The spin-echo EPI sequence included 31 slices covering the whole brain with echo time (TE) = 70 msec, time to repetition (TR) = 3200 msec, flip angle = 90°, 96 × 96 matrix, reconstructed in-plane resolution 2.5 × 2.5 mm, slice thickness 4.0 mm 896 images were acquired in total, collected in two runs of 24 min each. Following the standard method for distortion-corrected spin-echo fMRI (Embleton et al., 2010), the images were acquired with a single direction k space traversal

and a left-right phase encoding direction. In between the two functional runs, a brief “pre-scan” was acquired, consisting of 10 volumes of dual direction k space traversal SE EPI scans. This gave 10 pairs of images matching ATR activation the functional time series but with distortions in both phase encoding directions (10 left-right and 10 right-left). These scans were used in the distortion correction procedure. In addition, a high resolution T1-weighted 3D turbo field echo inversion recovery image was acquired (TR = 8400 msec, TE = 3.9 msec, flip angle 8°, 256 × 205 matrix reconstructed to 256 × 256, reconstructed resolution .938 × .938 mm, and slice thickness of 0.9 mm, SENSE factor = 2.5) with 160 slices covering the whole brain. This image was used for spatial normalisation. The spatial remapping Inositol oxygenase correction was computed using the method reported by Embleton

et al. (2010). In the first step, each image from the main functional time-series was registered to the mean of the pre-scan images using a 6-parameter rigid-body transformation in SPM8. Subsequently, a spatial transformation matrix was calculated from the pre-scan images, consisting of the spatial re-mapping necessary to correct the distortion. This transformation was then applied to each of the 896 co-registered functional images. Analysis was carried out using SPM8. The motion and distortion-corrected images for each participant were first co-registered to their T1 structural scan. Spatial normalisation of the T1 scans into MNI space was computed using DARTEL (Ashburner, 2007) and the resulting transformation applied to the functional images, which were resampled to 2 × 2 × 2 mm voxel size and smoothed with an 8 mm FWHM Gaussian kernel. At this point, temporal signal-to-noise (TSNR) maps were generated for each participant by dividing the mean signal in each voxel by its standard deviation (Murphy, Bodurka, & Bandettini, 2007). The mean TSNR map across all participants is shown in Fig. 1. TSNR exceeded 80 in ventral temporal regions.

Additionally, we found that HIF-1α overexpression diminished VEGF production, whereas only AdHIF-2α transduction resulted in elevation of VEGF expression. Therefore, it seems that two isoforms of HIF may play a distinct role in regulation of VEGF production in porcine proximal tubular epithelial cells, which are the major target of OTA action. Moreover, only HIF-2 exerts protective effect, especially against short-term acute kidney injuries. These results are in accordance with studies showing that HIF

may be protective in acute renal injuries whether in case of chronic ones they exert opposite effect (Manotham et al., 2004). Still, the role of each HIF Romidepsin purchase isoform in different kidney cell types may be various. Additionally, also the other factors, such as AP-1 and SP-1, should be investigated in this context. In conclusion, we have shown complicated pattern of VEGF regulation by different toxins affecting kidney biology. To our knowledge, the influence of AAI and OTA on some transcription factors have not been investigated before and further investigations are necessary to analyze this intriguing effects. The author declares that there are no conflicts of interest. This work was supported by grants from Polish Ministry for Science and Higher Education (Nos.: N N401 297835 and N N301 033440). The Faculty of Biochemistry, Biophysics and Biotechnology of the Jagiellonian

University is a IGF-1R inhibitor beneficiary of the structural funds from the Clomifene European Union and the Polish Ministry of Science and Higher Education (Grants Nos.: POIG.02.01.00-12 064/08, POIG 01.01.02-00-109/09, POIG.02.02.00-014/08 and 01.01.02-00-069/09). A.J. is a recipient of the Wellcome Trust International Senior Research Fellowship in Biomedical Science. A.L. is a recipient of Fellowship for Young Scientists funded by Ministry of Science and Higher Education. “
“Fluoxetine (FLX) is a selective serotonin reuptake inhibitors (SSRIs) with controversial

effects on carcinogenesis, that was reported to be ineffective against aggressive T-cell lymphoma in nude athymic mice, despite the significant decrease of such tumors in BALB/c mice, in which it possibly acted on immune system to inhibit tumor growth (Frick et al., 2008). However, it has been shown to enhance apoptosis and control cell cycle in Burkitt lymphoma, in spite of not affecting the viability of non-tumor peripheral blood mononuclear cells (Serafeim et al., 2003). Meanwhile, FLX has been reported to promote metastasis formation in young transplanted melanoma mice (Kubera et al., 2009). Once FLX is orally administered, it has a direct contact with the epithelia in the gastrointestinal tract (Arimochi and Morita, 2006), inducing an increase of serotonin (5-HT) levels by the blockade of serotonin reuptake transporter (SERT) (Bertrand et al., 2008).

The labeled cRNAs were

purified (QIAquick spin columns, QIAGEN, Venlo, The Netherlands) and 1 μg of each sample was hybridized to 4 × 44 K whole genome mouse oligo microarrays (G4122F, Agilent) according to manufacturer’s instructions (two-color microarray-based gene expression analysis, Agilent). After a 17-h incubation CHIR99021 period, slides were washed using various dilutions of SSPE (sodium chloride, sodium phosphate, EDTA) buffer according to the protocol provided by Agilent. Arrays were scanned using an Agilent microarray scanner (G2565B). The fluorescent readings from the scanner were converted to quantitative files using Feature Extraction 9.1 software (Agilent Technologies). Quality check of the arrays was done using software package LimmaGUI

in R version 2.3.1. Four samples were removed from the analysis due to technical failure. Data were imported in GeneMaths XT 1.5 (Applied Maths, St. MartensLatem, Belgium), and spots with signal intensities below two times PARP inhibitor background were excluded from subsequent analysis. Corrected data were normalized and adjusted for random and systematic error (Pellis et al., 2003). Significance analysis of microarrays (SAM) analysis was applied to detect significantly affected genes for each treatment using the two-class unpaired comparison (Tusher et al., 2001). The False Discovery Rate was set to < 0.5%. No additional filtering on a threshold for up- or downregulation was applied. Evaluation of the outcome of the SAM results showed that the minimal ratio for up- or downregulation was 1.5. Hierarchical clustering was done oxyclozanide with the programs Cluster (uncentered correlation; average linkage

clustering) and Treeview (Eisen et al., 1998). Metacore (GeneGo, St. Joseph, MI) is an online software program that provides, among other options, pathway analysis of microarray data. Groups of co-clustering genes were analyzed for overrepresentation of genes from signaling and metabolic pathways based on hypergeometric distribution (Ekins et al., 2006). Pathways with a p value < 10−5 were considered significant. Gene set enrichment analysis (GSEA) was performed to discover the differential expression of biologically relevant sets of genes that share common biological function or regulation (Subramanian et al., 2005). GSEA has the advantage that no initial filtering is applied to the data set to select for significantly differentially expressed genes. GSEA first ranks all probe sets based on fold changes (algorithm signal to noise) in expression between a treatment and the control. Subsequently, by using pre-defined sets of associated genes based on prior biological knowledge, GSEA calculates whether sets as a whole are enriched at the top or bottom of the fold change-based ranking list, or randomly distributed (Subramanian et al., 2005).

Additional http://www.selleckchem.com/products/PLX-4032.html information on patient characteristics is summarized in Supplementary Tables S1, S2, and S3. Frozen tumor samples were homogenized using a bead mill (TissueLyser, Qiagen) and tissue protein extraction reagent (T-PER, Thermo Scientific) supplemented with 1 mM EDTA, 5 mM NaF, 2 µM staurosporine, PhosSTOP Phosphatase Inhibitor Cocktail (Roche Applied Science), and Complete Mini Protease Inhibitor Cocktail (Roche Applied Science). Total protein concentration was determined by bicinchoninic acid assay (Thermo Scientific). Prior to spotting, tumor lysates were mixed with 4× SDS sample buffer (10% glycerol,

4% SDS, 10 mM DTT, 125 mM Tris–HCl, pH 6.8) and boiled for 5 min at 95 °C. Tumor lysates (total protein concentration 2 µg/µl) and dilution series of tumor sample pools serving as controls were spotted as technical triplicates and four identical subarrays on nitrocellulose-coated glass slides (Oncyte Avid, Grace-Biolabs) using a contact spotter (Aushon BioSystems). Slides were blocked with blocking buffer for fluorescent

applications (Rockland Immunochemicals) in TBS (50%, v/v) containing 5 mM NaF and 1 mM Na3VO4 Akt inhibitor for 2 h at RT, prior to incubation with target-specific primary antibodies at 4 °C over night (Supplementary Table S4). Primary antibodies (n = 128) were selected to recognize proteins involved in major cancer signaling pathways with a special focus on breast cancer biology. Only highly target-specific antibodies

were used and their validation was carried out as previously described [ 20]. Detection of primary antibodies was done with Alexa Fluor 680 F(ab′)2 fragments of goat anti-mouse IgG or anti-rabbit IgG in 1:8000 dilution (Life Technologies). In addition, representative slides were stained for total protein quantification using the protein dye Fast Green FCF as described Rucaparib in vivo before [ 21]. Images of all slides were obtained at an excitation wavelength of 685 nm and a resolution of 21 µm using the Odyssey Scanner (LI-COR). Signal intensities of each individual spot were quantified using GenePixPro 5.0 (Molecular Devices). Data preprocessing and quality control were performed with the R-package RPPanalyzer [ 22]. RPPA data of the discovery and the test cohort have been deposited in NCBI’s Gene Expression Omnibus [ 23] and are accessible through GEO series accession number GSE47066 and GSE50861, respectively. We set up a biomarker (feature) selection workflow including three different algorithms for classification (SCAD-SVM: support vector machines using smoothly clipped absolute deviation penalty; RF-Boruta: random forests using the Boruta algorithm for feature selection; PAM: prediction analysis for microarrays utilizing the nearest shrunken centroid classifier [[24], [25] and [26]]). We implemented the software in the R programming language and made it available through the bootfs R-package (https://r-forge.r-project.org/projects/bootfs/).

Fogarty et al. already demonstrated the

effect of short segments of empathy to decrease psychological arousal in clinical communication [6]. Our study further elaborates on this finding by showing that a few empathic remarks also have the power to affect physiological activity of APs’ SNS. These insights might be valuable to clinicians. Firstly, activation of the SNS is known to influence patients’ well-being [1]. Secondly, the effect of a core aspect of clinical communication, conveying medical information [52], can be severely hampered LY2109761 purchase due to the effect of SNS activation on patients’ memory [18]. As expected from prior research (e.g. [28]), affective communication did not only affect AP’s physiological arousal, but also improved APs’ recall of provided information, potentially partly by reducing physiological arousal. Notably, recall was only improved for information that was provided during the part of the consultation where the clinician HDAC inhibitor used affective communication and physiological arousal was lowered; 21% of the variance in recall could be explained by variance in physiological arousal. This might be an indication that patients’ psychophysiological responses to clinicians’ communication play a mediating role in the effectiveness of affective communication, more specifically in improving recall. Although we have not tested the connection between physiological arousal and recall

directly, our results illustrate the often emphasised importance of addressing patients’ emotions in clinical communication [52] and suggest that clinicians need to deal with patients’ emotions before providing additional

medical information to them. The strength of this study is the use of an experimental design, which allowed us to investigate the causal effect of communication in a bad news consultation. Another strength is the measurement of physiological arousal [50], since it offered the opportunity to get a better understanding of the mechanisms underlying patients’ cognitive and Thiamet G emotional processes during bad news consultations. Last, it allowed us to investigate the effects of specific communication elements more objectively and in different parts of the consultation [31] and [44]. The study also has some limitations. Although the analogue patient paradigm allowed us to use an experimental design, it might lowered the ecological validity of the results, as our results are based on findings from healthy participants, not clinical patients. Although a recent review study demonstrated that using APs do seem to be valid [41], clinical patients might react differently. However, in case of real bad news consultations, physiological responses might even be stronger and information recall further hampered, thus enhancing the potential alleviating role of affective communication. This has to be tested in clinical studies.

Adsorption is also the process employed for the removal of phenylalanine from protein hydrolysates in the preparation of Phe-free dietary formulas for treatment of phenylketonuria patients (Clark, Alves, Franca, & Oliveira, 2012). Many studies have been reported in the scientific literature dealing with the adsorption of phenylalanine on materials such as activated carbons, zeolites, Venetoclax cell line ion exchangers, polymeric resins and others (Díez et al., 1998, Titus et al., 2003, Garnier et al., 2007, Piao et al., 2009, Ghosh et al., 2011 and Fei-Peng et al., 2012). However, high costs are associated with the production and regeneration of such adsorbents

and these costs could be reduced by the use of low-cost adsorbents (Clark et al., 2012). Agricultural wastes are the most common raw materials being studied as potential precursors for the preparation of low-cost adsorbents, since they are renewable, available in large amounts and potentially less expensive than other

materials to manufacture a diversity of adsorbents. Brazil is the third largest corn producer in the world, with an expected production of almost 52 million tons in 2012. Solid residues from corn production such as corn cobs present great potential for use as raw materials in the production of adsorbents (Bagheri & Abedi, 2011). Reports on the use of agricultural residues for Phe removal from aqueous solutions by adsorbents based on agricultural click here Forskolin residues are not available in the literature, with the exception of our previous study employing defective coffee bean press cake (Clark et al., 2012). Changes in the precursor material significantly modify the physico-chemical characteristics of the adsorbing surfaces and thus significantly affect the types of adsorbate–adsorbent interactions, which, in the case of phenylalanine, range from hydrogen bonding to Coulombic to hydrophobic interactions.

Hence, studies of adsorption of phenylalanine onto residue-based adsorbents will contribute to a better understanding of the adsorption of this type of amino acid on such materials and also provide theoretical guides for the implementation of practical processes such as separation or purification of this amino acid. In view of the aforementioned, the objective of this work was to evaluate the performance of corn cobs in the production of adsorbents for Phe removal from aqueous solutions. Corn cobs were provided by EMBRAPA (Sete Lagoas, Brazil). The phenylalanine standard and other reagents were purchased from Sigma–Aldrich (SP, Brazil). The adsorbent was prepared according to the procedure employed in a previous study (Clark et al., 2012) using coffee press cake as a precursor material (3 min impregnation with H3PO4 followed by 1 h activation in a muffle furnace). The activated material was cooled under nitrogen and washed with distilled water until constant pH = 6, dried at 105 °C for 24 h and ground (Arbel grinder, São Paulo, Brazil, 0.15 < D < 1.00 mm).

10.1 statistical package® (The R Foundation for Statistical

Computing, Vienna, Austria) to obtain general prevalence and 95% confidence intervals estimated. After parasitological examination, regardless of infection status, all persons were treated with a standard dose of praziquantel (40 mg/kg) (ShinPoong Pharma., Seoul, Republic of Korea) and a single 400 mg tablet of albendazole (GSK, London, UK), or a half tablet for children aged under two years. On the basis of a positive blood film, or Paracheck© test, non-pregnant women and children were offered Lonart (20 mg/120 mg artemether/lumefrantrine medication; Cipla, Mumbai, India) while pregnant women were offered quinine sulphate tablets (Zest Pharma, Madhya Pradesh, India), as supervised by the project nurse click here and monitored the following day. A total of 15 GPS-data loggers (I-GotU GT-120, Mobile Action, UK) were available for this study. After completing a brief baseline acceptance survey questionnaire, mothers selected at random were requested

to carry this small unit (dimension 44.5 x 28.5 x 13 mm, weight 20 g) back to their homestead, returning it to the field medical team the same day. The unit was powered by a rechargeable 230 mAh Lithium-ion battery which, if set for GPS-data logging at 3-minute intervals, lasts for up to three days before needing recharging. The units were ‘locked’ electronically to avoid any external tampering. Upon receipt of the unit, data were offloaded onto a personal computer onsite as GPX files which were then used directly in GoogleEarth 5 (Google Inc., CA, USA) and ArcView 9.3 (ESRI, CA, USA) GIS. Using NVP-BEZ235 chemical structure the log it was possible to ascertain, more easily, the position of the homestead. Whilst identity records were kept anonymous, the infection status of each mother and child was used to annotate the maps to reveal any micro-patterning. To investigate the positional accuracy of the I-GotU device, the lead author accompanied 15 mothers back to their household whilst carrying a Garmin Oregon 550t handheld unit (Garmin, KS, USA). These track logs were later downloaded and directly compared against those obtained from

the I-GotU. To identify clustering of infection, Staurosporine price a spatial scan statistic (Satscan v9.1) was performed.23 Based on an expectation of Poisson distribution of cases of infection amongst all possible subject locations surveyed, the spatial scan statistic considered whether the number of cases in an area was excessively high or low. The scan consisted of placing circles of varying radius distances centred at each subject’s household location, and computing ratios of observed to expected cases. Both clusters of high and low prevalence were searched for in the scan. The scan statistic was performed separately for schistosomiasis, hookworm, and malaria prevalence. Additionally, a scan was performed for multiple parasite infection, i.e., persons with more than one type of parasite infection.

The saltern lies about 500 m from the Mediterranean Sea in the north. It consists of a series of shallow ponds with depths of 0.5–1.5 m and surface areas varying from 70 to a few hundred ha (Figure 1). Seawater is pumped from the Suez Canal through an intake to a large pond (P1) where solar energy and wind combine and evaporation begins. The water volume is reduced and salinity levels gradually build up through consecutive evaporation ponds (P2–P3) and the production pond (P4). As the salinity increases, low-soluble salts precipitate

AZD6738 as carbonates and sulphates. The saturated brine then passes through smaller ponds (P5, crystallizer ponds) where evaporation continues (Figure 2). Once the volume has been reduced to about 10% of the original, any furtherk concentration results in the deposition of sodium chloride. Five ponds with different salinities were sampled in summer (June 2010).

Water samples were collected 20 cm below the surface using a 2-L Van Dorn bottle. Water temperature, transparency and pH were measured immediately in situ after sampling using a mercury SB431542 research buy glass thermometer graduated in 0.1 °C, a Secchi disc and a portable pH meter (Model HI 9124) respectively. Salinity was estimated as total dissolved salts (TDS) according to APHA (1995). A well-mixed sample was passed through a glass fibre filter, after which the filtrate was evaporated to dryness in a weighed

dish, then dried to constant weight at 180 °C. The increase in dish weight represents the salt content [g l− 1]. The total weight of major ions generally Adenosine triphosphate constitutes over 99% of the total salinity (Wetzel & Likens 2000). Nitrates and phosphates were determined in filtered seawater using GF/C filters according to the methods described by Parsons et al. (1984). For phytoplankton examination, water samples were preserved immediately using Lugol’s iodine and concentrated by decanting. The algal count was conducted under an inverted microscope using Utermöhl’s method (Utermöhl 1958) and identified to genus or species level by consulting the works of Prescott (1951), Hendey (1964), Dodge (1982) and Komárek & Anagnostidis (2005). Pearson’s correlation coefficient was performed using the SPSS 17 software program to examine the potential relationships among physicochemical variables and phytoplankton diversity and density. Relations highly significant to the 0.05 level were noted. The waters of the Port Fouad saltworks were always clear, with the Secchi depth corresponding to the maximum depth of water due to the shallowness of the ponds (Table 1). The water of the shallower, more saline pond (P5, crystallizer pond) was warmer (29.3 °C) than that of the deeper, less saline pond (P1, 25.6 °C). The water salinity increased progressively throughout the series of interconnected ponds, giving a value of 340.