Composition of the oil-slime microbial community as determined by analysis of the 16S rRNA gene

Analysis of the 16S rRNA genes of the cultured microorganisms of industrial oil-slime revealed predominance (~85-90%) of the Gammaproteobacteria in the community of aerobic heterotrophs and specific oil-slime degraders. Relation of the isolated strains with members of the genera Pseudomonas, Stenotrophomonas, and Enterobacter was established. Analysis of the same gene in the total DNA from the oil-slime revealed greater microbial diversity (~20 operative taxonomic units determined by T-RFLP) than in the cultured part of the community, which included ∼12 different colony types. Three major restriction fragments were found, with their total area ∼50%. These results demonstrated the low morphological and phylogenetic diversity of the oil-slime bacterial community. © 2013 Pleiades Publishing, Ltd

Full-scale bioreactor pretreatment of highly toxic wastewater from styrene and propylene oxide production

The wastewater originating from simultaneous production of styrene and propylene oxide (SPO) is classified as highly polluted with chemical oxygen demand level in the range 5965 to 9137mgL-1-as well as highly toxic. The dilution factor providing for a 10 percent toxic effect of wastewater samples in a test with Paramecium caudatum was 8.0-9.5. Biological approach for pretreatment and detoxification of the wastewater under full-scale bioreactor conditions was investigated. The number of suspended microorganisms and the clean up efficiency were increased up to 5.5-6.58×108CFUmL-1 and 88 percent, respectively during the bioreactor's operation. Isolates in the Citrobacter, Burkholderia, Pseudomonas, and Paracoccus genera were dominant in the mature suspended, as well as the immobilized microbial community of the bioreactor. The most dominant representatives were tested for their ability to biodegrade the major components of the SPO wastewater and evidence of their role in the treatment process was demonstrated. The investigated pretreatment process allowed the wastewater to be detoxified for conventional treatment with activated sludge and was closely related to the maturation of the bioreactor's microbial community. © 2014 Elsevier Inc

Preparations of Bacillus pumilus secreted RNase: One enzyme or two?

© 2015, Pleiades Publishing, Ltd. Immunochemical analysis of the following purified preparations of Bacillus pumilus RNase (binase) was carried out: industrially manufactured enzyme (Institute of Organic Synthesis, Riga, Latvia) and the enzymes isolated from the culture liquid of the native B. pumilus producer and from the Escherichia coli BL21 recombinant strain bearing the pGEMGX1/ent/Bi plasmid. Electrophoresis of all three samples of purified binase revealed two protein fractions with ribonuclease activity possessing molecular masses of ∼12 and 25 kDa. The possible presence of binase II, a second secreted RNase, was ruled out. Both high- and low-molecular mass proteins interacted with binase-specific antibodies in the immunoblotting reaction, which indicated their antigenic identity. The difference in molecular mass between these proteins indicated the possible presence of two forms of binase in solution, a monomer and a dimer

Cytokine Levels in the Serum of Patients with Chronic Kidney Insufficiency Before and After Hemodialysis

© 2016, Springer Science+Business Media New York.Chronic kidney insufficiency (CKI) is often the end point of a broad range of chronic kidney diseases and characterized with decreasing number of functionally active nephrons. Pathophysiological CKI is characterized by decreased glomerular filtration, which leads to accumulation of life-threatening toxic metabolites. Hemodialysis is the main therapeutic measure aimed to prolong patient’s life until kidney transplant is available. The goal of this study is to analyze serum level of 21 cytokines in CKI. We have found that the serum level of several (IL-2Rα, IL-3, IL-12 (p40), IL-16, IL-18, HGF, MIF, CSF-1, MCP-3, CXCL12, SCF, IFN-α2, LIF, β-NGF, and CXCL1) cytokines and chemokines was upregulated in CKI without hemodialysis as compared to controls (p = 0.005). Interestingly, serum cytokines were also upregulated in serum of CKI patients who received hemodialysis. Upregulated cytokines are associated with inflammation and activation of Th1 lymphocytes. We suggest that hemodialysis has limited effect on serum cytokine levels. It could be concluded that therapeutic effect of hemodialysis is not associated with removal of inflammatory cytokines from circulation. Further studies will help better define the underlying cause of an increased inflammation in CKI and identify the laboratory criteria for anti-inflammatory therapy

Competitive ability of <i>Escherichia coli</i> strains in the intestinal microbiota of patients with Crohn's disease and healthy volunteers: physiological, biochemical and genetic characteristics

Introduction. Crohn's disease (CD) is a chronic inflammation of various parts of the gastrointestinal tract with an increased proportion of Escherichia coli. However, the role of E. coli in disease remains unclear. This study aims to evaluate the competitive abilities of E. coli strains from CD patients and healthy volunteers, and to identify the biochemical and genetic determinants underlying these features. Materials and methods. The antagonistic activity was assessed by co-cultivation of 11 clinical E. coli strains inhibiting the growth of the K-12, with Enterobacter cloacae, Klebsiella pneumonia and Salmonella enterica. To elucidate the mechanism of antagonistic activity, the evaluation of biochemical properties and a comparative genomic analysis were used. Results and discussion. Genes of bacteriocin production systems were identified in genomes of 11 strains from CD patients and healthy volunteers active against the E. coli K-12 strain. Three strains from healthy individuals demonstrated activity against several Enterobacteriaceae bacteria. The strains biochemical properties were typical of representatives of E. coli. Strains 1_34_12, active against E. cloacae, and 1_45_11, inhibiting all tested enterobacteria, are phylogenetically related to the laboratory strain K-12. Strain 1_39_1, active against K. pneumonia and S. enterica, is phylogenetically close to the Nissle1917, contains the genes for colibactin biosynthesis and a variant of the fimH gene that increases the adhesive ability of bacteria. Conclusion. The identified E. coli strains are able to displace Enterobacteriaceae bacteria and can be used to study the bacteria-bacteria and host-bacteria interactions, to understand their role in gut homeostasis and intestinal inflammation

Self-assembled nanoformulation of methylprednisolone succinatewith carboxylated block copolymer for local glucocorticoid therapy

A new self-assembled formulation of methylprednisolone succinate (MPS) based on a carboxylatedtrifunctional block copolymer of ethylene oxide and propylene oxide (TBC-COOH) was developed. TBC-COOH and MPS associated spontaneously at increased concentrations in aqueous solutions to form almostmonodisperse mixed micelles (TBC-COOH/MPS) with a hydrodynamic diameter of 19.6 nm, zeta potentialof −27.8 mV and optimal weight ratio ∼1:6.3. Conditions for the effective formation of TBC-COOH/MPSwere elucidated by comparing copolymers and glucocorticoids with different structure. The micellarstructure of TBC-COOH/MPS persisted upon dilution, temperature fluctuations and interaction with bloodserum components. TBC-COOH increased antiradical activity of MPS and promoted its intrinsic cytotoxi-city in vitro attributed to enhanced cellular availability of the mixed micelles. Intracellular transportationand hydrolysis of MPS were analyzed using optimized liquid chromatography tandem mass spectrometrywith multiple reaction monitoring which showed increased level of both MPS and methylprednisolonein neuronal cells treated with the formulated glucocorticoid. Our results identify TBC-COOH/MPS as anadvanced in situ prepared nanoformulation and encourage its further investigation for a potential localglucocorticoid therapy

LC-MS Method Development for Simultaneous Determination of Trans-3′-hydroxycotinine and Three Mercapturic Acids in Urine

© 2018, Springer Science+Business Media, LLC, part of Springer Nature. The negative impact of tobacco smoke on the human body is due to a wide range of harmful substances including volatile organic compounds (VOCs). Some VOCs of tobacco smoke metabolize in human organism into mercapturic acids (MAs). The determination of the amount of MAs in readily available biological fluids, for example in urine, allows to assess the level of exposure of these VOCs in a particular person. It is useful to assess the impact of individual VOCs on the body together with the assessment of the intake of nicotine. The intake of nicotine can be determined by the content of its metabolites in the urine, in particular by the content of trans-3′-hydroxycotinine (tH-Cot). A joint assessment of the concentrations of trans-3′-hydroxycotinine and MAs in urine allows obtaining selective information about effects of different VOCs and nicotine on the smoker’s body. We have developed a liquid chromatography–mass spectrometry (LC-MS) method for simultaneous quantifying of tH-Cot and three MAs: N-Acetyl-S-(3-hydroxypropyl)cysteine (HPMA), N-Acetyl-S-(3-hydroxypropyl-1-methyl)-L-cysteine (HMPMA), N-Acetyl-S-(2-cyanoethyl)-L-cysteine (CEMA). We used this method to quantify the levels of MAs and tH-Cot in the urine of a group of 15 smokers just before and 5 days after smoking cessation. For all studied compounds, we have found statistically significant changes in concentration on the fifth day of smoking cessation. The method developed can be used to jointly assess the levels of exposure to nicotine and VOCs in the study of various tobacco products

Characterization of Dysbiotic Changes of Skin Microbiota in Contact Sports Athletes

© 2017, Springer Science+Business Media New York.Contact sports athletes often suffer from various skin disorders (inflammatory diseases of bacterial and fungal origin, atopic dermatitis, psoriasis, etc.) resulting in long breaks in training which ruin athletic performance. Wrestling implies intense skin-to-skin contact that creates perfect conditions for transmission of the infectious agents. Following the standard rules of hygiene (showering and handwashing directly after each competition and training) does not exclude the possibility to get an infection from sparring partner. To characterize the skin microbial composition of wrestlers who do not have current manifestation of any skin disorders, the metagenomic analysis was performed. Absolute predominance of Bacillus genus in metagenomic profiles of wrestlers’ skin was observed in contrast with the existing literature data. Classic microbiological approaches allowed to detect hemolytic forms of microorganisms. Wrestlers’ skin appeared to be colonized with hemolytic bacilli, whereas the non-wrestler athletes did not have such bacteria on their skin. Such dysbiotic shifts in the microbial community may cause the emergence of skin diseases. Revealed properties could help to design highly effective antiseptics for the contact sports hygiene