Use of sawdust for production of ligninolytic enzymes by white-rot fungi and pharmaceutical removal.

Use of white-rot fungi for enzyme-based bioremediation of wastewater is of high interest. These fungi produce considerable amounts of extracellular ligninolytic enzymes during solid-state fermentation on lignocellulosic materials such as straw and sawdust. We used pure sawdust colonized by Pleurotus ostreatus, Trametes versicolor, and Ganoderma lucidum for extraction of ligninolytic enzymes in aqueous suspension. Crude enzyme suspensions of the three fungi, with laccase activity range 12-43 U/L and manganese peroxidase activity range 5-55 U/L, were evaluated for degradation of 11 selected pharmaceuticals spiked at environmentally relevant concentrations. Sulfamethoxazole was removed significantly in all treatments. The crude enzyme suspension from P. ostreatus achieved degradation of wider range of pharmaceuticals when the enzyme activity was increased. Brief homogenization of the colonized sawdust was also observed to be favorable, resulting in significant reductions after a short exposure of 5 min. The highest reduction was observed for sulfamethoxazole which was reduced by 84% compared to an autoclaved control without enzyme activity and for trimethoprim which was reduced by 60%. The compounds metoprolol, lidocaine, and venlafaxine were reduced by approximately 30% compared to the control. Overall, this study confirmed the potential of low-cost lignocellulosic material as a substrate for production of enzymes from white-rot fungi. However, monitoring over time in bioreactors revealed a rapid decrease in enzymatic ligninolytic activity.

Production of oyster mushroom (Pleurotus ostreatus) on sawdust supplemented with anaerobic digestate.

Anaerobic digestion of organic waste results in production of biogas and a nutrient-rich digestate that has an established use as fertilizer in plant production. This study evaluated use of anaerobic digestate based on a high concentration of organic household waste as a fertilizer in sawdust-based production of oyster mushrooms (Pleurotus ostreatus). Inclusion of 0.5 L of anaerobic digestate (AD) per kg sawdust gave similar productivity in terms of biological efficiency (79.5 ± 5.4 %), and protein concentration (24.7 ± 2.4 % of dry weight (dw)) as standard mushroom substrate (78.1 ± 5.3 %, and 21.9 ± 3.0 % of dw, respectively). However, mushroom growth was impaired at the highest concentration of anaerobic digestate tested, 1 L digestate per kg dw sawdust. Comparison of the AD-fertilized substrate with a mushroom substrate with standard components (sawdust, wheat bran, calcium sulfate) and with similar C/N-ratio revealed some differences in elemental composition of the fruiting bodies, with an major increase in sodium concentration for the AD-fertilized substrate compared with the standard substrate (413.3 ± 28.9 and 226.7 ± 30.6 mg kg-1 dw, respectively). This difference can be explained by high sodium concentration in the anaerobic digestate, most likely due to inclusion of food scraps from households and restaurants in the biodigester feedstock. Screening of both substrates for a total of 133 micropollutants revealed that total sum of micropollutants was significantly higher in the AD-fertilized substrate (258 ± 12 ng/g dw substrate) than in the standard substrate (191 ± 35 ng/g dw substrate). Nitrogen losses during preparation of the AD-fertilized substrate were negligible.

Use of lignocellulosic substrate colonized by oyster mushroom (Pleurotus ostreatus) for removal of organic micropollutants from water.

Efficient removal techniques are urgently needed to remove organic micropollutants (OMPs) from wastewater, in order to protect water resources. In this study, laccase activity of mushroom substrate colonized by Pleurotus ostreatus was evaluated as a novel wastewater treatment method for removal of OMPs, including diclofenac, bicalutamide, lamotrigine, and metformin at environmentally relevant concentrations. Laccase activity of the colonized mushroom substrate was found to be highest, 0.8 enzyme activity (U)/g mushroom substrate wet weight, immediately before initiation of fruiting body formation. The selected OMPs were treated for 5 min with suspensions of mushroom substrate with laccase activity of approximately 50 U/L. Removal of all OMPs was significant, with the highest removal for diclofenac of 90% compared with a control with uncolonized mushroom substrate. To our knowledge, direct use of colonized mushroom substrate in removing diclofenac from water has not been reported previously. Removal efficiency of bicalutamide, lamotrigine, and metformin was 43%, 73%, and 59%, respectively. This demonstrates potential for using mushroom substrate colonized by P. ostreatus for removal of OMPs from wastewater.

nPAsym: an open-source plugin for ImageJ to quantify nuclear shape asymmetry.

BACKGROUND AND OBJECTIVES: The nucleus is a complex and dynamic organelle enclosing the major part of the cell's genome. A growing body of evidence suggests that changes in the shape of this organelle can influence cell activities. The other way around, altered nuclear shape may be indicative of impaired cell function. Symmetry is an important aspect of nuclear shape not receiving the attention it merits. We address this problem by presenting a software tool allowing to quantify nuclear shape asymmetry in light microscopy images. METHODS: The software named nPAsym is written in Scala and implemented as a plugin to ImageJ making possible to use it in combination with other ImageJ tools. The plugin works with 8-bit images segmented into black nuclear masks and white background. It performs a number of operations allowing to analyze multiple objects within a single image, removing some segmentation artefacts, filtering out objects incomplete and below a specified size. The feature of interest is quantified using the notion of point asymmetry. The performance of nPAsym was tested in a small-scale study comparing nuclear shapes for cells of nodular goiter, follicular thyroid adenoma and papillary thyroid carcinoma. RESULTS: We present nPAsym, the ImageJ plugin, that measures nuclear shape asymmetry. It works with digital microscopic images segmented using either a raster graphics editor or built-in ImageJ functions. nPAsym is packaged in a single .jar file and does not require installation as well as configuration. It has proved effective in distinguishing between some of the nuclear shape phenotypes. CONCLUSIONS: nPAsym is the user-friendly, platform-independent and open-source software tool allowing to quantify nuclear shape asymmetry in digital images captured from cytologic and histologic preparations. It has a potential to become useful for both experimental research and diagnostics.

A passive sampling method for detecting analgesics, psycholeptics, antidepressants and illicit drugs in aquatic environments in the Czech Republic.

The goal of this study was to assess the bioavailable concentrations of analgesics, psycholeptics, antidepressants and illicit drugs in the surface waters of the Czech Republic. All of the sampling sites are located within the most important water quality monitoring profiles at the Czech Hydrometeorological Institute. The total concentrations of the compounds ranged from 463 to 6,447 ng POCIS(-1) (Polar Organic Chemical Integrative Sampler). Carbamazepine (196-2,690 ng POCIS(-1)) and tramadol (160-2,250 ng POCIS(-1)) were the most abundant compounds at every site. The most polluted sites were those that received communal wastewater effluent and had a low dilution factor (ratio of wastewater effluent and river flow). The aqueous concentrations of the target compounds were estimated using sampling rate values obtained during a field calibration experiment. Patterns in the aqueous concentrations of the compounds (after back calculation from POCIS extracts) and the POCIS concentrations are different, possibly leading to discrepancies between the toxicity assessments conducted using POCIS extracts and those conducted using grab samples of water from the same location.

Calcidiol and prostate cancer.

Epidemiological studies suggest that serum calcidiol (25(OH)-Vitamin D3) seems to be associated with several cancers including prostate cancer. We have made several experimental studies in order to clarify the mechanism(s) involved in the association. Calcidiol has been regarded as an inactive prohormone for calcitriol, which possesses the highest biological activity of the Vitamin D metabolites, when it is evaluated on the basis of bioactivity/nmol. However, we found recently that at the physiological concentration calcidiol (100-200 nM) is an active hormone, whereas calcitriol (1alpha,25(OH)2-Vitamin D3) (100 pM) is inactive in human primary prostate stromal cells. Calcidiol is able to inhibit cell growth and to induce or inhibit several genes including 1alpha-hydroxylase and 24-hydroxylase genes. This suggests that calcidiol might be an independent endocrine system involved in the control of cell differentiation and proliferation, whereas calcitriol might be mainly involved in the regulation of calcium and phosphorous balance. Several mechanisms may mediate the action of Vitamin D in the prostate. This is a review of some recent studies on the role of (1) Vitamin D metabolism, (2) growth factors and (3) fatty acid metabolism.

C-fos and IL-2 gene expression in rat brain cells and splenic lymphocytes after nonantigenic and antigenic stimuli.

Immunostimulatory or immunosuppressive stress models were used: (1) rotation stress (RS) and (2) immobilization (restraint) stress (IS). Intravenous injection of tetanus toxoid (anatoxin) (TT) was chosen as the antigenic stimulus (500 micrograms/kg weight), and intravenous injection of saline solution was used as the control. Splenic lymphocytes (CBA mice) or different brain structures (Wistar and Sprague-Dawley rats) were analyzed. The c-fos and interleukin-2 (IL-2) mRNA expression was measured using a digoxigenin (Dig)-labeled cDNA probe by spot or in situ hybridization. Rotation stress stimulated IL-2 mRNA synthesis in lymphocytes in the presence of ConA and rIL-2 by 40%. IL-2 mRNA synthesis in lymphoid cells obtained from animals after IS and after IS in combination with the administration in vitro of the cytotoxic drug CsA to the splenic lymphocytes was inhibited (30% and 99%), accordingly, as compared with control rats. Induction of c-fos mRNA synthesis in rat brain cells was noted 30 minutes after RS in the hypothalamus (lateralis hypothalamic area, LHA), thalamus, corpus collosum, and sensorimotor zone of the brain cortex. IL-2 mRNA synthesis was shown two hours after RS in the same structures. The increased number of c-fos mRNA-positive cells two hours after TT injection was shown in the posterior hypothalamus area (PHA), LHA, dorsomedial nucleus (DMH), ventromedial nucleus (VMH), and anterior hypothalamus area (AHA) as compared to the effect of i.v. saline injection. Moreover, IL-2 mRNA-positive cell induction was noted in the PHA, DMH, and VMH. Six hours after TT injection, c-fos mRNA expression was decreased in the PHA, LHA, and AHA. Activation of c-fos and IL-2 mRNA was detected in the paraventricularis nucleus 6 hours after TT i.v. injection. Thus, inhibition or stimulation of IL-2 gene expression in lymphoid cells depends on the nature of the stressors. RS or antigenic stimuli induce c-fos and IL-2 gene expression in definite structures of the brain. The dynamics of this process are time dependent. The partial correlation between c-fos and IL-2 mRNA expression in localization in brain structures and time dependence was shown.

[Molecular-biologic aspects of interaction between nervous and immune systems]. / Molekuliarno-biologicheskie aspekty vzaimodeistviia nervnoi i immunnoi sistem.

The problem of the neuro-immuno interactions on the level of the protein trans-factors, stimulating interleukin-2 (IL-2) gene expression was discussed. The physico-chemical and functional parameters of the low molecular nuclear proteins (SP and BP- 14, 18, 19 kDs) isolated from splenic and brain cells of immunized rats were studied. The binding of these proteins to the regulatory region of IL-2 gene in vitro and stimulation of the IL-2mRNA synthesis in splenic T-lymphocytes culture in normal conditions were shown. The protective effect of SP and BP on the IL-2mRNA synthesis in stressful conditions and by the T-cells treatment with the CsA was demonstrated.

[Study of reproductive behavior in Russian and Byelarus populations exposed to ionizing radiation as a consequence of the Chernobyl AES accident]. / Izuchenie reproduktivnogo povedeniia v populiatsiiakh Rossii i Belorussii, podvergshikhsia vozdeistviiu ioniziruiushchego izlucheniia v rezul'tate avarii na Chernobyl'skoi AES.

As a result of the Chernobyl accident the population in Gomel and Mogilev districts of Belorussia as well as in Bryansk district in Russia were exposed to ionizing radiation. The changes in pregnancy outcomes (spontaneous abortions, stillbirths, etc) are the integral indices reflecting the state of the population under post-accident conditions. The data obtained don't allow us to isolate reliably the radiation-induced component in post-accident behavior of the studied populations.

Transactivation of interleukin-2 gene via the nuclear proteins from spleen and brain cells.

Immunomodulatory action of nuclear proteins derived from spleen and brain of immunized rats was found in ConA-stimulated mouse spleen lymphocyte cultures. In vitro experiments on 32P- or Dig-labeled promoter-enhancer region (-548 bp) of the human interleukin-2 (IL-2) gene showed specific DNA-nuclear protein complex formation for spleen (6-14 kD) and brain (21 kD) proteins. The functional influence of these trans-factors on the IL-2 promoter region was studied in the model system of Jurkat cells with the firefly luciferase (Luc) reporter gene. The spleen and brain proteins were shown to influence the membrane surface properties of thymic and spleen cells as revealed in a two-phase aqueous system. We conclude that both spleen and brain from immunized rats contain certain protein trans-factors which can be active in T cells and display a common mode of regulatory action the level of IL-2 gene.

[Immunomodulating peptides, regulation of interleukin-2 expression and its possible mechanism]. / Immunomoduliruiushchie peptidy, reguliatsiia obrazovaniia interleikina-2 i vozmozhnyi ee mekhanizm.

The influence of some immunomodulation peptides SKD, R and L on the interleukin-2 (IL-2) synthesis in T-lymphocytes and possibility of their specific interaction with nucleotide sequences of IL-2 gene were studied. It was firstly shown that peptides R and L specifically interacted with regulatory nucleotide sequences of IL-2 gene and, possibly, play a key role in the immunomodulation actions on the IL-2 production.