Microfluorometric filter determination of DNA in sucrose density gradients

A simple filter method for the fluorometric estimation of DNA in alkaline and neutral sucrose gradient fractions with DABA·2HCl is discussed. Alpha-450 membrane filters of regenerated cellulose (Gelman Instrument Co., Ann Arbor, Michigan) were used. In the proposed method washing of the DNA precipitate as well as the reaction with DABA·2HCl were performed directly on the filters, thus avoiding repeated washing and centrifugations of DNA precipitates applied hitherto in analogous fluorometric techniques. A good coincidence of the results concerning localization of DNA sedimentation profiles determined by radioisotopic and fluorometric methods was obtained. The method is very convenient for DNA estimation in alkaline and neutral sucrose density gradient fractions obtained by ultracentrifugation of DNA of nonproliferating cells where DNA labeling is very difficult, and in the case of human lymphocytes even impossible without stimulation for blastic transformation. Its other advantages are a considerably simplified procedure and a higher precision with respect to other fluorometric methods for determination of DNA.     

Uptake of immune RNA by normal mouse spleen cells

Summary Normal mouse spleen cells take up in vitro radioactively labeled immune RNA. RNA taken up is present in nuclei, polysomes, membranes and cytoplasm. About 20–40% of immune RNA is nonspecifically associated with cell surface. 45% of RNA taken up is degraded and reutilized inside the cells within 2 hours.This work was supported by the Polish Academy of Sciences within the project 09.7.4.1.1.     

Transformation of Curvularia lunata IM 2901 with pAN7-1 influences selected physiological properties of the fungus

Genetic analysis of Curvularia lunata IM 2901 transformants, previously obtained by electroporation with plasmid pAN7-1, was carried out. Isolates displayed several differences in hygromycin B resistance and their physiology. It was shown that plasmid pAN7-1 was integrated in different copy numbers and at different positions in the genome of the strains studied. Both the wild type and pAN7-1 isolates, when growing in liquid media, produced an extracellular emulsifying agent. The transformants differed in their growth kinetics, intensity of surfactant production and in the efficiency of cortexolone 11beta-hydroxylation, in comparison with the wild type. The micro-organisms varied in susceptibility to the lytic enzyme complex (Novozyme 234), which indicated the presence of differences in their cell wall composition and/or in architecture caused by an integrated plasmid pAN7-1.     

Biotransformation of 4‐fluoro‐N‐(1‐{2‐[(propan‐2‐yl)phenoxy]ethyl}‐8‐azabicyclo[3.2.1]octan‐3‐yl)‐benzenesulfonamide,a novel potent 5‐HT7 receptor antagonist with antidepressant‐like and anxiolytic properties: In vitro and in silico approach

The aim of the study was to investigate the metabolism of 4‐fluoro‐N‐(1‐{2‐[(propan‐2‐yl)phenoxy]ethyl}‐8‐azabicyclo[3.2.1]octan‐3‐yl)‐benzenesulfonamide (PZ‐1150), a novel 5‐HT₇ receptor antagonist with antidepressant‐like and anxiolytic properties, by the following three ways: in vitro with microsomes; in vitro employing Cunninghamella echinulata, and in silico using MetaSite. Biotransformation of PZ‐1150 with microsomes resulted in five metabolites, while transformation with C. echinulata afforded two metabolites. In both models, the predominant metabolite occurred due to hydroxylation of benzene ring. In silico data coincide with in vitro experiments, as three MetaSite metabolites matched compounds identified in microsomal samples. In human liver microsomes PZ‐1150 exhibited in vitro half‐life of 64 min, with microsomal intrinsic clearance of 54.1 μL/min/mg and intrinsic clearance of 48.7 mL/min/kg. Therefore, PZ‐1150 is predicted to be a high‐clearance agent. The study demonstrated the applicability of using microsomal model coupled with microbial model to elucidate the metabolic pathways of compounds and comparison with in silico metabolite predictions.     

Macrophyte and macroinvertebrate patterns in unimpacted mountain rivers of two European ecoregions

Hydrobiologia - The aim of the study was to compare the patterns of development of macrophytes and macroinvertebrates in different types of reference mountain rivers. The study is based on...     

The Content of Elements in Rainwater and Its Relation to the Frequency of Hospitalization for Chronic Lymphocytic Leukemia and Chronic Myeloid Leukemia in Opole Voivodship, Poland, During 2000–2002

Environmental factors play an essential role in the etiology of diseases of the hematopoietic system. Such factors include soil and water pollution and the presence of metals and toxic compounds in the air. Measuring the content of metallic elements in rainwater has become an accepted procedure for environmental pollution monitoring. In accordance with the above, it was decided to study relations between the content of selected elements in rainwater and hospitalization frequency due to chronic lymphocytic leukemia (CLL, C91 on ICD-10) and chronic myeloid leukemia (CML, C92 on ICD-10). It can be assumed that hospitalization frequency is a reliable indicator of exacerbations of these diseases. The annual average of hospitalizations due to a given disease during the years 2000–2002 was correlated with the annual average content of a given element in rainwater using the Spearman’s correlation indicator to describe the relationship between the element content and the disease that is possibly a consequence of the element’s presence in rainwater. In cases of CLL for all the subjected population and for men, no statistically significant correlations were found. For women, statistically significant correlations were found for chromium (r?=?0.66), lead (r?=?0.58), copper (r?=?0.58), and cadmium (r?=?0.51). For CML in all the studied population significant, negative correlations were found for magnesium (r?=??0.6) and zinc (r?=?-0.52). In men, significant negative correlations were seen for magnesium (r?=??0.69 and zinc (r?=??0.55). No significant correlations were found in women. These results indicate the need of taking into account the environmental and gender factors in research connected with these diseases, which can be probably of help in improvements of therapy efficiency.     

The mycobiota of landfill leachates in the pretreatment process in a sequencing batch reactor

The paper discusses the dynamics of the accumulation of microscopic fungi, depending on the sludge load (Bx), in activated sludge used for landfill leachate pretreatment. The propagule washout from the sludge into pretreated leachates is determined, including genera and species that may threaten environmental health. An increased accumulation of microscopic fungi in sludge flocs occurred at Bx=0.23−0.45 mg chemical oxygen demand (COD) mg⁻¹ d⁻¹. Microscopic fungi were eluted at the maximal Bx value tested of 1.64 mg COD mg⁻¹ d⁻¹. Both the activated sludge and the leachate runoff from the sequencing batch reactor (SBR) pose health risks to the environment due to the occurrence of fungi such as Aspergillus fumigatus, Purpureocillium lilacinum, Cyberlindnera jadinii (C. utilis), Geotrichum candidum and G. fragrans. Their count is sufficient to cause multi-organ infections in homeothermal animals and in humans.