Inactivation of Heparin by Cationically Modified Chitosan

This study was performed to evaluate the ability of N-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride (HTCC), the cationically modified chitosan, to form biologically inactive complexes with unfractionated heparin and thereby blocking its anticoagulant activity. Experiments were carried out in rats in vivo and in vitro using the activated partial thromboplastin time (APTT) and prothrombin time (PT) tests for evaluation of heparin anticoagulant activity. For the first time we have found that HTCC effectively neutralizes anticoagulant action of heparin in rat blood in vitro as well as in rats in vivo. The effect of HTCC on suppression of heparin activity is dose-dependent and its efficacy can be comparable to that of protamine-the only agent used in clinic for heparin neutralization. HTCC administered i.v. alone had no direct effect on any of the coagulation tests used. The potential adverse effects of HTCC were further explored using rat experimental model of acute toxicity. When administered i.p. at high doses (250 and 500 mg/kg body weight), HTCC induced some significant dose-dependent structural abnormalities in the liver. However, when HTCC was administered at low doses, comparable to those used for neutralization of anticoagulant effect of heparin, no histopathological abnormalities in liver were observed.     

Studies on Survival of Horse Oocytes After Rapid-i Method Vitrification

The aim of the present study was to determine the vitality and developmental competence of equine oocytes after in vitro maturation (IVM) and vitrified by Rapid-i method. In experiment 1, oocytes after IVM were vitrified using media: EquiPro VitKit (group 1) or medium containing 18% Ficoll, 40% ethylene glycol, and 0.3 M sucrose (group 2). For evaluation of toxicity effect, oocytes were exposed to media without a plug to liquid nitrogen. To evaluate viability, oocytes were stained with fluorescein diacetate and ethidium bromide. In experiment 2, oocytes after IVM and vitrification were activated by 7.5 μM ionomicin in TCM 199 (5 minutes) combined with 2 mM 6-DMAP in TCM 199 with 10% fetal bovine serum (4.5 hours). Survival rate was: 63% in group 1 (n = 54), 55% in group 2 (n = 69), and 73.2% (n = 56) in the control group. After parthenogenetic activation, 10.2% (n = 49) of 2–4 blastomeres were observed. This percentage was lower than in the nonvitrified group: 38.5% (n = 53).     

The influence of \upbeta-hydroxy- \upbeta-methylbutyrate (HMB) on cell-mediated immunity in tench Tinca tinca (L.): in vitro and in vivo study

$\upbeta$ -Hydroxy- $\upbeta$ -methyl butyrate (HMB) has been shown to counteract many of the negative effects of intensive fish production methods and results in increased growth and protection against diseases. In the present study, the in vitro and in vivo effect of HMB on the immunocompetence cell activity in tench Tinca tinca (L.) was examined. In the in vitro study pronephric phagocytes and lymphocytes were isolated from the fish and grown in culture medium (RPMI-1640) containing 0, 0.1, 1, 5, 10, 25, 50 or $100\,\upmu$ g HMB cm^?3 of medium. The effects of HMB on the respiratory burst activity (RBA), the potential killing activity (PKA) and lymphocyte proliferation stimulated by concanavalin A (ConA) or lipopolysaccharide (LPS) were examined. The in vitro study showed that HMB increased the RBA and PKA of the phagocytes, compared to the medium over that of cells grown without HMB. Lymphocyte proliferation stimulated by ConA and LPS was also increased approximately when HMB was added to the culture medium at concentrations between 10 and $100\,\upmu$ g HMB cm^?3. In the in vivo study fish were fed daily with pellets containing HMB at doses 0, 10, 25 and 50 mg kg^?1 body weight day^?1. The study showed that HMB statistically increased the RBA and PKA and highest activity at 50 mg kg^?1 body weight day^?1 were observed. Also lymphocyte proliferation stimulated by ConA and LPS were maximally stimulated at dose 50 mg kg^?1 body weight day^?1. In conclusion, the current study shows that HMB could potentially improve immunocompetence cell activity in tench through increased cell proliferation and functionality.     

Theory untangles the high-resolution infrared spectrum of the ortho-H2-CO van der Waals complex

Rovibrational spectroscopy of molecules boasts extremely high precision, but its usefulness relies on the assignment of spectral features to corresponding quantum mechanical transitions. In the case of ortho-H(2)-CO, a weakly bound complex abundant in the interstellar medium (although not yet observed there), the rather complex spectrum has been unexplained for more than a decade. We assigned this spectrum by comparison with a purely ab initio calculation. For most lines, agreement to within 0.01 centimeter(-1) between experiment and theory was achieved. Our results show that the applicability of rovibrational spectroscopy can be extended with the assistance of high-accuracy quantum mechanical computations.     

In vitro efficacies of phosphorolytic enzymes synthesized in mycelial cells of Aspergillus niger AbZ4 grown by a liquid surface fermentation

Activities of phytase, a pH 6.0 optimum nonspecific phosphomonoesterase and phosphodiesterase assayed toward bis(p-nitrophenyl)phosphate (phosphodiesterase I) and against p-nitrophenylphosphorylcholine (phosphodiesterase II), were partially purified from mycelial extracts of Aspergillus niger AbZ4 cultivated on a molasses medium by a liquid surface fermentation method. After elimination of phosphate from the medium, 7.3- and 3.5-fold enhancements in specific activities of phytase and phosphodiesterase II were observed. Efficacies of mycelial protein fractions in dephosphorylating a wheat-based broiler feed were determined in vitro according to a procedure that simulated digestion in the intestinal tract of poultry. The addition of 0.052 mg of protein from fractions, each of which was high in either pH 6.0 optimum phosphomonoesterase, phosphodiesterase I, phosphodiesterase II, or phytase per gram of a feed sample resulted in the enhancement of phosphorus release by 10, 11, 27, and 88%, respectively. In the presence of an excess of commercial phytase, the addition of the mycelial fraction high in phytase increased the dephosphorylation rate by 56%. The fraction high in phosphodiesterase II enhanced feed dephosphorylation by 8% in the presence of an excess of commercial phytase and commercial acid phosphatase.     

Design and operation of drainage-subirrigation systems in Poland

The different techniques used in the design and operation of drainage-subirrigation systems in low-lying riverine areas in Poland are presented. The required groundwater levels used as designing criteria and the applications of the steady state and unsteady state approach to ditch (drain) spacing design in different soil conditions are discussed. The practical application of groundwater level maintenance using the techniques of controlled drainage, subirrigation with a constant water level, and subirrigation with a regulated water level, are shown for three different field sites.     

The effect of the source and dosage of dietary Cu on redox status in rat tissues

The aim of this experiment was to investigate whether the amount of Cu added to the diet of rats can be reduced without adversely affecting the antioxidant status of tissues and growth, and whether copper nanoparticles can be used for this purpose. For four weeks, four experimental groups of rats were fed diets with two dosages of added Cu (standard—6.5 or 3.25 mg/kg) in two forms (standard—CuCO₃ or copper nanoparticles). Replacing the CuCO₃ supplement with CuNPs resulted in a decreased lung weight and an increased Cu content in brain, kidney and lung, intensification of lipid peroxidation processes, and weakened antioxidant defence in the lungs and kidneys. This treatment also reduced the Cu content in heart, level of lipid oxidation in the liver and testes and improved antioxidant defence in the brain. Reducing the addition of Cu to the diet from 6.5 to 3.25 mg/kg reduced lung weight and increased lipid peroxidation in the liver, heart and lungs, and also weakened antioxidant defence in the lungs and testes. This treatment also weakened the lipid peroxidation process in the spleen, small intestine and brain and strengthened the antioxidant defence of the brain and kidneys. In conclusion, replacing CuCO₃ with CuNPs and reducing the level of Cu in the diet of rats has a particularly unfavourable effect on the respiratory system, causing adverse changes in the lungs. However, these treatments have a clearly positive effect on the redox status of the liver and brain.     

Polymorphism analysis of housekeeping genes for identification and differentiation of <Emphasis Type="Italic">Clavibacter michiganensis</Emphasis> subspecies

The utility of polymorphism analysis was determined for differentiation of the following subspecies of the Gram-positive plant pathogenic bacterium, Clavibacter michiganensis: C. m. subsp. michiganensis, C. m. subsp. sepedonicus, C. m. subsp. insidiosus C. m. subsp. nebraskensis, and C. m. subsp. tessellarius. Specific primers designed for amplification of the housekeeping genes recA, rpoB, and rpoD generated 827-, 1037-, and 862-bp DNA fragments, respectively. PCR products obtained from 40 C. michiganensis strains were analysed using RFLP with four restriction endonucleases, and those PCR products with specific RFLP patterns were sequenced. The genotypes discriminated after PCR–RFLP were specific for each subspecies and also allowed for differentiation of C. m. subsp. michiganensis strains. Sequence analysis of the recA, rpoB, and rpoD gene fragments also distinguished C. michiganensis subspecies and was useful for phylogenetic analysis of all subspecies. For rapid, inexpensive, and effective differentiation of the five subspecies in this research, we recommend the amplification of recA and/or rpoD gene fragments and digestion of the PCR products with the restriction endonuclease FnuDII.     

Electrochemically mediated atom transfer radical polymerization

Atom transfer radical polymerization is a versatile technique for exerting precise control over polymer molecular weights, molecular weight distributions, and complex architectures. Here, we show that an externally applied electrochemical potential can reversibly activate the copper catalyst for this process by a one-electron reduction of an initially added air-stable cupric species (Cu(II)/Ligand). Modulation of polymerization kinetics is thereby tunable in real time by varying the magnitude of applied potential. Application of multistep intermittent potentials successfully triggers initiation of polymerization and subsequently toggles the polymerization between dormant and active states in a living manner. Catalyst concentrations down to 50 parts per million are demonstrated to maintain polymerization control manifested in linear first-order kinetics, a linear increase in polymer molecular weight with monomer conversion, and narrow polymer molecular weight distributions over a range of applied potentials.     

Distribution of polycyclic aromatic hydrocarbons (PAHs) in the bottom sediments of a dam reservoir,their interaction with organic matter and risk to benthic fauna

Journal of Soils and Sediments - The aims of the study were to investigate the interaction between fractions of organic matter and polycyclic aromatic hydrocarbons (PAHs) in bottom sediments and to...