Dynamic adaptation of the extremophilic red microalga Cyanidioschyzon merolae to high nickel stress.

The order of Cyanidiales comprises seven acido-thermophilic red microalgal species thriving in hot springs of volcanic origin characterized by extremely low pH, moderately high temperatures and the presence of high concentrations of sulphites and heavy metals that are prohibitive for most other organisms. Little is known about the physiological processes underlying the long-term adaptation of these extremophiles to such hostile environments. Here, we investigated the long-term adaptive responses of a red microalga Cyanidioschyzon merolae, a representative of Cyanidiales, to extremely high nickel concentrations. By the comprehensive physiological, microscopic and elemental analyses we dissected the key physiological processes underlying the long-term adaptation of this model extremophile to high Ni exposure. These include: (i) prevention of significant Ni accumulation inside the cells; (ii) activation of the photoprotective response of non-photochemical quenching; (iii) significant changes of the chloroplast ultrastructure associated with the formation of prolamellar bodies and plastoglobuli together with loosening of the thylakoid membranes; (iv) activation of ROS amelioration machinery; and (v) maintaining the efficient respiratory chain functionality. The dynamically regulated processes identified in this study are discussed in the context of the mechanisms driving the remarkable adaptability of C. merolae to extremely high Ni levels exceeding by several orders of magnitude those found in the natural environment of the microalga. The processes identified in this study provide a solid basis for the future investigation of the specific molecular components and pathways involved in the adaptation of Cyanidiales to the extremely high Ni concentrations.

Breastmilk mineral composition among well-educated mothers from Central Poland - Associations with maternal dietary intake, dietary patterns and infant psychomotor development.

BACKGROUND: Maternal dietary habits could affect breastmilk mineral composition, which may influence infant development. Mineral dietary intake or supplementation slightly affects its breastmilk concentration. However, the intake of selected food groups or dietary patterns that reflect diet complexity could have a greater impact. Hence, the aim of the study was to assess breastmilk mineral composition at one, three, and six months of lactation among mothers living in urban area of Central Poland, as well as the evaluate maternal dietary determinants and associations with infant anthropometric and psychomotor development. METHODS: The study was conducted among 43 healthy and exclusively breastfeeding mothers. In the first, third, and sixth months of lactation, we collected breastmilk samples and assessed the concentration of Ca, P, Zn, Fe, Se, Ni, As, Pb, and Cd using the ICP-MS method. Maternal dietary habits were evaluated by a food frequency questionnaire in the first month of lactation, whereas in the third and sixth by the three-day food record. Based on the collected data adherence to the Polish-adapted Mediterranean (Pl-aMED; 1 month) and the DASH diet (Mellen's Index; 3 and 6 months) was assessed. In the third and sixth months of lactation infant anthropometric parameters and the sixth month of lactation psychomotor development were evaluated. RESULTS: Breastmilk Se, Ni, As, Pb, and Cd levels were under the LOQ in all the breastmilk samples at all study visits. Median breastmilk mineral concentrations of Ca, P, Zn, and Fe in the first, third, and sixth months of lactation varied from 381.9 to 332.7 mg/L, 161.6 to 139.1 mg/L, 2.2 to 0.8 mg/L, and 0.26 to 0.17 mg/L, respectively. Maternal dietary intake and supplementation did not affect breastmilk Ca, P, Zn, and Fe. Pl-aMED scores were associated with breastmilk Ca (ß = 0.489, 95% CI 0.180 - 0.799, p = 0.003) and Zn (ß = 0.499, 95% CI 0.199 - 0.798, p = 0.002) in the first month of lactation, whereas no association with the DASH diet were observed in the third and sixth month of lactation. Breastmilk Fe in the third month was associated with infant motor development (ß = 0.420, 95% CI 0.113 - 0.727, p = 0.009) in the sixth month of life, but no other associations with anthropometric or psychomotor development were observed. Moreover, we estimated that few infants meet their adequate intake (AI) requirements for P, Zn, and Fe. CONCLUSION: Our study showed that maternal adherence to Pl-aMED is a significant predictor of breastmilk Ca and Zn in the first month of lactation, which may be especially important considering that more than 75% of infants had inadequate Zn intake. Moreover, we found that breastmilk Fe positively influenced infant motor development, despite the majority of infants having inadequate intake. On the other hand, no infant had deficiency symptoms, which emphasizes the necessity to evaluate of AI norms for infants.

Replicated, urban-driven exposure to metallic trace elements in two passerines.

While there are increasing examples of phenotypic and genotypic differences between urban and non-urban populations of plants and animals, few studies identified the mechanisms explaining those dissimilarities. The characterization of the urban landscape, which can only be achieved by measuring variability in relevant environmental factors within and between cities, is a keystone prerequisite to understand the effects of urbanization on wildlife. Here, we measured variation in bird exposure to metal pollution within 8 replicated urbanization gradients and within 2 flagship bird species in urban evolutionary ecology: the blue tit (Cyanistes caeruleus) and the great tit (Parus major). We report on a highly significant, positive linear relationship between the magnitude of urbanization-inferred as either tree cover, impervious surface cover, or an urbanization score computed from several environmental variables, and copper, zinc and lead concentrations in bird feathers. The reverse relationship was measured in the case of mercury, while cadmium and arsenic did not vary in response to the urbanization level. This result, replicated across multiple cities and two passerine species, strongly suggests that copper, zinc, lead and mercury pollution is likely to trigger the emergence of parallel responses at the phenotypic and/or genotypic level between urban environments worldwide.

Searching for Low Molecular Weight Seleno-Compounds in Sprouts by Mass Spectrometry.

A fit for purpose analytical protocol was designed towards searching for low molecular weight seleno-compounds in sprouts. Complementary analytical techniques were used to collect information enabling the characterization of selenium speciation. Conceiving the overall characterization of the behavior of selenium, inductively plasma optical mass spectrometry (ICP-MS) was used to determine the total selenium content in entire sprouts as well as in selected extracts or chromatographic fractions. Then, high-performance liquid chromatography combined with ICP-MS (HPLC-ICP-MS) was used to evaluate the presence of inorganic and organic seleno-compounds, with the advantages of being very sensitive towards selenium, but limited by available selenium standard compounds. Finally, ultra-high performance liquid chromatography electrospray ionization triple quadrupole mass spectrometry (UHPLC-ESI-QqQ-MS/MS) and UHPLC-ESI-Orbitrap-MS/MS were used for the confirmation of the identity of selected compounds and identification of several unknown compounds of selenium in vegetable sprouts (sunflower, onion, radish), respectively. Cultivation of plants was designed to supplement sprouts with selenium by using solutions of selenium (IV) at the concentration of 10, 20, 40, and 60 mg/L. The applied methodology allowed to justify that vegetable sprouts metabolize inorganic selenium to a number of organic derivatives, such as seleno-methylselenocysteine (SeMetSeCys), selenomethionine (SeMet), 5'-seleno-adenosine, 2,3-DHP-selenolanthionine, Se-S conjugate of cysteine-selenoglutathione, 2,3-DHP-selenocysteine-cysteine, 2,3-DHP-selenocysteine-cysteinealanine, glutathione-2,3-DHP-selenocysteine, gamma-Glu-MetSeCys or glutamyl-glycinyl-N-2,3-DHP-selenocysteine.

Urinary metabolomic signature of muscle-invasive bladder cancer: A multiplatform approach.

Bladder cancer (BCa) is ninth amongst the most common types of cancer in the human population worldwide. The statistics of incidence and mortality of BCa are alarming and the currently applied diagnostic methods are still not sensitive enough. This leads to a large number of undiagnosed BCa cases, usually among patients in the early stages of the disease. Despite the fact that many risk factors of BCa have been recognized, the pathomechanism of development of bladder cancer has not been fully explained yet. Therefore, in the present study, multiplatform urinary metabolomics has been implemented in order to scrutinize potential diagnostic indicators of BCa that might help to explain its pathomechanism and be potentially useful in diagnosis and determination of stage of the disease. Urine samples collected from muscle-invasive high grade BCa patients (n = 24) and healthy volunteers (n = 24) were matched in terms of most common BCa risk factors i.e. gender, age, BMI and smoking status. They were analyzed by high performance liquid chromatography coupled with time of flight mass spectrometry detection (HPLC-TOF/MS) using RP and HILIC chromatography, gas chromatography hyphenated with triple quadruple mass spectrometry detection (GC-QqQ/MS) in scan mode, and proton nuclear magnetic resonance (1H NMR). The six datasets obtained were submitted to univariate and multivariate statistical analyses. 17 metabolites significantly discriminated urinary profiles of BCa patients from urinary profiles of healthy volunteers. These metabolites are mainly involved in amino acid metabolism, pyrimidine and purine metabolism, as well as energy metabolism and might play a crucial role in the pathogenesis of BCa.

Effect of carbonate substitution on physicochemical and biological properties of silver containing hydroxyapatites.

Ag+-substituted hydroxyapatites (Ag-HAs) and Ag+/CO32--co-substituted hydroxyapatites (Ag-CHAs) with two different concentrations of silver ions were synthesized by the standard precipitation method. For comparison, pure hydroxyapatite (HA) and carbonated hydroxyapatite (CHA) were synthesized using the same method. The obtained powders were examined by various physicochemical methods, such PXRD, TEM, FTIR and ssNMR. Elemental analysis was provided by WD-XRF and ICP-MS methods. The strains of Staphylococcus aureus and Escherichia coli were used to evaluate the antibacterial activity of the materials. The study demonstrates that the substituted samples are homogenous and poorly crystalline. Introducing carbonates into the crystal structure significantly affects the physicochemical properties of the silver containing hydroxyapatite, i.e., crystallinity, lattice parameters, crystal size and morphology or content of structural hydroxyl groups. Carbonate substitution leads to a significant increase in the contribution of the hydrated surface layer. Ag-CHA nanocrystals with an enhanced hydrated surface layer and higher solubility demonstrate an admirable antibacterial effect.

Binding and Conversion of Selenium in Candida utilis ATCC 9950 Yeasts in Bioreactor Culture.

Selenium is considered an essential component of all living organisms. The use of yeasts as a selenium supplement in human nutrition has gained much interest over the last decade. The accumulation and biochemical transformation of selenium in yeast cells is particularly interesting to many researchers. In this article, we present the results of the determination of selenium and selenomethionine content in the biomass of feed yeast Candida utilis ATCC 9950 obtained from the culture grown in a bioreactor. The results indicated that C. utilis cells performed the biotransformation of inorganic selenium(IV) to organic derivatives (e.g., selenomethionine). Selenium introduced (20-30 mg Se4+∙L-1) to the experimental media in the form of sodium(IV) selenite (Na2SeO3) salt caused a significant increase in selenium content in the biomass of C. utilis,irrespective of the concentration. The highest amount of selenium (1841 µg∙gd.w.-1) was obtained after a 48-h culture in media containing 30 mg Se4+∙L-1. The highest content of selenomethionine (238.8 µg∙gd.w.-1) was found after 48-h culture from the experimental medium that was supplemented with selenium at a concentration of 20 mg Se4+∙L-1. Biomass cell in the cultures supplemented with selenium ranged from 1.5 to 14.1 g∙L-1. The results of this study indicate that yeast cell biomass of C. utilis enriched mainly with the organic forms of selenium can be a valuable source of protein. It creates the possibility of obtaining selenium biocomplexes that can be used in the production of protein-selenium dietary supplements for animals and humans.

Selol, an organic selenium donor, prevents lipopolysaccharide-induced oxidative stress and inflammatory reaction in the rat brain.

Neuroinflammation and oxidative stress are key intertwined pathological factors in many neurological, particularly neurodegenerative diseases, such as Alzheimer's and Parkinson's disorders as well as autism. The present study was conducted to evaluate the protective effects of Selol, an organic selenium donor, against lipopolysaccharide (LPS)-mediated inflammation in rat brain. The results demonstrated that the peripheral administration of LPS in a dose of 100 µg/kg b.w. evoked typical pathological reaction known as systemic inflammatory response. Moreover, we observed elevated blood levels of thiobarbituric acid-reactive substances (TBARS), a marker of oxidative stress, as well as increased concentration of tumor necrosis factor-α (TNF-α) in LPS-treated animals. Selol significantly prevented these LPS-evoked changes. Subsequently, Selol protected against LPS-induced up-regulation of proinflammatory cytokines (Tnfa, Ifng, Il6) in rat brain cortex. The molecular mechanisms through which Selol prevented the neuroinflammation were associated with the inhibition of oxidized glutathione (GSSG) accumulation and with an increase of glutathione-associated enzymes: glutathione peroxidase (Se-GPx), glutathione reductase (GR) as well as thioredoxin reductase (TrxR) activity and expression. Finally, we observed that Selol administration effectively protected against LPS-induced changes in the expression of brain-derived neurotrophic factor (Bdnf). In conclusion, our studies indicated that Selol effectively protects against LPS-induced neuroinflammation by inhibiting pro-inflammatory cytokine release, by boosting antioxidant systems, and by augmenting BDNF level. Therefore, Selol could be a multi-potent and effective drug useful in the treatment and prevention of brain disorders associated with neuroinflammation.

Bio-transformation of selenium in Se-enriched bacterial strains of Lactobacillus casei.

BACKGROUND: Selenium is an element of very great importance for the proper functioning of the human body, mainly due to its antioxidant properties. Selenium exhibits a preventive effect in the case of cardiovascular disease, the immune system, male infertility and inhibits the toxic action of other agents. Selenium is important for Hashimoto's disease. Intake of selenium in the diet slows the aging process. The biological and toxicological effects of selenium strongly depend on its chemical form. Some organisms for example: plant, yeast, are capable of metabolizing low bioavailable selenium compounds (inorganic selenium) into its high bioavailable forms (organic selenium). OBJECTIVE: The aim of this study was to investigate the bio-transformation of selenium by Lactobacillus bacteria towards the characterisation of selenium metabolites. MATERIAL AND METHODS: The speciation of selenium was evaluated by high performance liquid chromatography with inductively coupled plasma mass spectrometry detector. The extraction of selenium species from lyophilized bacteria was executed with water, the mixture of lipase and protease, as well as lisozyme and sodium dodecyl sulphate. RESULTS: All investigated bacteria strains cultivated in the presence of Na2SeO3 effectively uptake selenium. Surprisingly, none of the applied extraction media exhibited a strong power to release the majority of the uptaken selenium compounds. Thus a maximum of 10% of the selenium was extracted from bacteria exposed to the enzymes. However, it was found that Lactobacillus bacteria are able to metabolize inorganic ions of selenium (IV) into Se-methionine, Se-methyloselenocysteine and other unidentified forms. CONCLUSIONS: The study confirmed the ability of probiotic bacteria to biotransform inorganic selenium into its organic derivatives. Therefore, Se-enriched bacteria can be considered as an addition to the functional food. KEY WORDS: selenium speciation, extraction procedure, Lactobacillus casei bacteria, Lactic acid bacteria (LAB), HPLC ICP-MS, functional food.

Electrochemical examination of ability of dsDNA/PAM composites for storing and releasing of doxorubicin.

Composites consisting of ss- and ds-DNA strands and polyacrylamide (PAM) hydrogel have been synthesized. DNA was entrapped non-covalently. The obtained DNA biomaterial exhibited a strong increase in guanine and adenine anodic currents when temperature reached the physiological level. This increase was related to the unique oligonucleotide structural changes in the composite. The structural alterations in the PAM lattices were employed for the release of the drug accumulated in the composite. Doxorubicin (Dox) was selected as the drug; it was accumulated by intercalation to dsDNA and was slowly released from the dsDNA/PAM system by using a minor temperature increase (up to 40÷45 °C) as it is routinely done in hyperthermia. The applied release temperature was either constant or oscillating. The binding strength, the rate of Dox release and the properties of the composite were examined using voltammetry, SEM and ICP-MS.