Physiological and transcriptome profiling of Chlorella sorokiniana: A study on azo dye wastewater decolorization.

Over decades, synthetic dyes have become increasingly dominated by azo dyes posing a significant environmental risk due to their toxicity. Microalgae-based systems may offer an alternative for treatment of azo dye effluents to conventional physical-chemical methods. Here, microalgae were tested to decolorize industrial azo dye wastewater (ADW). Chlorella sorokiniana showed the highest decolorization efficiency in a preliminary screening test. Subsequently, the optimization of the experimental design resulted in 70% decolorization in a photobioreactor. Tolerance of this strain was evidenced using multiple approaches (growth and chlorophyll content assays, scanning electron microscopy (SEM), and antioxidant level measurements). Raman microspectroscopy was employed for the quantification of ADW-specific compounds accumulated by the microalgal biomass. Finally, RNA-seq revealed the transcriptome profile of C. sorokiniana exposed to ADW for 72 h. Activated DNA repair and primary metabolism provided sufficient energy for microalgal growth to overcome the adverse toxic conditions. Furthermore, several transporter genes, oxidoreductases-, and glycosyltransferases-encoding genes were upregulated to effectively sequestrate and detoxify the ADW. This work demonstrates the potential utilization of C. sorokiniana as a tolerant strain for industrial wastewater treatment, emphasizing the regulation of its molecular mechanisms to cope with unfavorable growth conditions.

Detection of pesticides in food products using paper-based devices by UV-induced fluorescence spectroscopy combined with molecularly imprinted polymers.

In this proof-of-concept study, we explore the detection of pesticides in food using a combined power of sensitive UV-induced fingerprint spectroscopy with selective capture by molecularly imprinted polymers (MIPs) and portable cost-effective paper-based analytical devices (PADs). The specific pesticides used herein as model compounds (both pure substances and their application products for spraying), were: strobilurins (i.e. trifloxystrobin), urea pesticides (rimsulfuron), pyrethroids (cypermethrine) and aryloxyphenoxyproponic acid herbicides (Haloxyfop-methyl). Commercially available spraying formulations containing the selected pesticides were positively identified by MIP-PADs swabs of sprayed apple and tomato. The key properties of MIP layer - imprinting factor (IF) and selectivity factor (α) were characterized using trifloxystrobin (IF-3.5, α-4.4) was demonstrated as a potential option for in-field application. The presented method may provide effective help with in-field testing of food and reveal problems such as false product labelling.

Evaluation of mercury bioavailability and phytoaccumulation by means of a DGT technique and of submerged aquatic plants in an aquatic ecosystem situated in the vicinity of a cinnabar mine.

The ability of submerged aquatic plants (Elodea canadensis, Myriophyllum spicatum, Ceratophyllum demersum) and a natant plant (Eichhornia crassipes) to bioaccumulate mercury was evaluated in a laboratory experiment as well as in a real aquatic ecosystem situated in the vicinity of a cinnabar mine. Moreover, the ability of the diffusive gradients in the thin films technique (DGT) to predict mercury bioavailability for selected aquatic plants was tested. The submerged plants had sufficient bioaccumulation capacity for long-term phytoaccumulation of mercury in a real aquatic ecosystem. The determined bioaccumulation factor was greater than 1000. On average, the submerged plant leaves accumulated 13 times more mercury than the leaves of the natant aquatic plants. Chlorides at concentrations up to 200 mg/L had no statistically significant effect on mercury accumulation, nevertheless, the presence of humic acid in the water environment resulted in its significant (p < 0.002) decrease. A strong positive correlation (r > 0.66) was determined between mercury concentration in the input parts (leaves and/or roots) of the aquatic plants and the flow of mercury into DGT units.

Prognostic Significance of Serum Free Amino Acids in Head and Neck Cancers.

Despite distinctive advances in the field of head and neck squamous cell cancer (HNSCC) biomarker discovery, the spectrum of clinically useful prognostic serum biomarkers is limited. As metabolic activities in highly proliferative transformed cells are fundamentally different from those in non-transformed cells, specific shifts in concentration of different metabolites may serve as diagnostic or prognostic markers. Blood amino acids have been identified as promising biomarkers in different cancers before, but little is known about this field in HNSCC. Blood amino acid profiles of 140 HNSCC patients were examined using high-performance liquid chromatography. Cox proportional hazards regression model was used to assess the prognostic value of amino acid concentrations in serum. Colony forming assay was used to identify the effect of amino acids that were significant in Cox proportional hazards regression models on colony forming ability of FaDu and Detroit 562 cell lines. In the multivariable Cox regression model for overall survival (OS), palliative treatment was associated with an unfavourable prognosis while high serum levels of methionine have had a positive prognostic impact. In the relapse-free survival (RFS) multivariable model, methionine was similarly identified as a positive prognostic factor, along with tumor localization in the oropharynx. Oral cavity localization and primary radio(chemo)therapy treatment strategy have been linked to poorer RFS. 1mM serine was shown to support the forming of colonies in both tested HNSCC cell lines. Effect of methionine was exactly the opposite.

Separation, identification and quantification of carotenoids and chlorophylls in dietary supplements containing Chlorella vulgaris and Spirulina platensis using High Performance Thin Layer Chromatography.

In this study, 14 commercial products (dietary supplements) containing alga Chlorella vulgaris and cyanobacteria Spirulina platensis, originated from China and Japan, were analysed. UV-vis spectrophotometric method was applied for rapid determination of chlorophylls, carotenoids and pheophytins; as degradation products of chlorophylls. High Performance Thin-Layer Chromatography (HPTLC) was used for effective separation of these compounds, and also Atomic Absorption Spectrometry for determination of heavy metals as indicator of environmental pollution. Based on the results obtained from UV-vis spectrophotometric determination of photosynthetic pigments (chlorophylls and carotenoids), it was confirmed that Chlorella vulgaris contains more of all these pigments compared to the cyanobacteria Spirulina platensis. The fastest mobility compound identified in Chlorella vulgaris and Spirulina platensis using HPTLC method was ß-carotene. Spectral analysis and standard calibration curve method were used for identification and quantification of separated substances on Thin-Layer Chromatographic plate. Quantification of copper (Cu2+, at 324.7 nm) and zinc (Zn2+, at 213.9nm) was performed using Flame Atomic Absorption Spectrometry with air-acetylene flame atomization. Quantification of cadmium (Cd2+, at 228.8 nm), nickel (Ni2+, at 232.0nm) and lead (Pb2+, at 283.3nm) by Electrothermal Graphite Furnace Atomic Absorption Spectrometry; and quantification of mercury (Hg2+, at 254nm) by Cold Vapour Atomic Absorption Spectrometry.

Toxicity of naturally-contaminated manganese soil to selected crops.

The impact of manganese excess using naturally contaminated soil (Mn-soil, pseudototal Mn 6494 vs 675 µg g(-1) DW in control soil) in the shoots of four crops was studied. Mn content decreased in the order Brassica napus > Hordeum vulgare > Zea mays > Triticum aestivum. Growth was strongly depressed just in Brassica (containing 13696 µg Mn g(-1) DW). Some essential metals (Zn, Fe) increased in Mn-cultured Brassica and Zea, while macronutrients (K, Ca, Mg) decreased in almost all species. Toxic metals (Ni and Cd) were rather elevated in Mn-soil. Microscopy of ROS, NO, lipid peroxidation, and thiols revealed stimulation in all Mn-cultured crops, but changes were less visible in Triticum, a species with low shoot Mn (2363 µg g(-1) DW). Antioxidative enzyme activities were typically enhanced in Mn-cultured plants. Soluble phenols increased in Brassica only while proteins rather decreased in response to Mn excess. Inorganic anions (chloride, sulfate, and phosphate) were less accumulated in almost all Mn-cultured crops, while the nitrate level rather increased. Organic anions (malate, citrate, oxalate, acetate, and formate) decreased or remained unaffected in response to Mn-soil culture in Brassica, Hordeum, and Triticum but not in Zea. However, the role of organic acids in Mn uptake in these species is not assumed. Because control and Mn-soil differed in pH (6.5 and 3.7), we further studied its impact on Mn uptake in solution culture (using Mn concentration ∼5 mM deducted from water-soluble fraction of Mn-soil). Shoot Mn contents in Mn-treated plants were similar to those observed in soil culture (high in Brassica and low in Triticum) and pH had negligible impact. Fluorescence indicator of "general ROS" revealed no extensive or pH-dependent impact either in control or Mn-cultured roots. Observed toxicity of Mn excess to common crops urges for selection of cultivars with higher tolerance.

Capillary zone electrophoretic determination of phenolic compounds in chess (Bromus inermis L.) plant extracts.

A simple CZE method for quantification of phenolic compounds (vanillin, cinnamic, sinapic, chlorogenic, syringic, ferulic, benzoic, p-coumaric, vanillic, p-hydroxybenzoic, rosmarinic, caffeic, gallic and protocatechuic acids) in less than 10 min using 20 mM sodium tetraborate (pH 9.2) with 5% v/v methanol as a BGE and with UV detection at 254 nm is described. The LODs (3 S/N) ranged between 0.02 and 0.12 microg/ mL. Repeatabilities (RSDs) were 0.66-1.8 and 1.56-4.23% for migration times and peak areas (n = 5), respectively. The method was applied to the determination of phenolic compounds in chess (Bromus inermis L.) after Soxhlet extraction and purification of the crude extracts with SPE procedures. The results compared well with those obtained by liquid chromatographic method. B. inermis was found as a suitable model plant containing a broad spectrum of phenolic compounds in easily detectable concentrations and as a potential source of antioxidants.

Separation of phenolic acids by capillary electrophoresis with indirect contactless conductometric detection.

A new method for the electrophoretic separation of nine phenolic acids (derivatives of benzoic and cinnamic acids) with contactless conductometric detection is presented. Based on theoretical calculations, in which the mobility of the electrolyte co- and counterions and mobility of analytes are taken into consideration, the electrolyte composition and detection mode was selected. This approach was found to be especially valuable for optimization of the electrolyte composition for the separation of analytes having medium mobility. Indirect conductometric detection mode was superior to the direct mode as predicted theoretically. The best performance was achieved with 150 mM 2-amino-2-methylpropanol electrolyte at pH 11.6. The separation was carried out in a counter-electroosmotic mode and completed in less than 6 min. The LODs achieved were about 2.3-3.3 microM and could be further improved to 0.12-0.17 microM by using a sample stacking procedure. The method compares well to the UV-Vis detection.