Glutamine synthetase content and immunochemical analysis of different strains of Bordetella pertussis

In this investigation 3 groups of strains isolated from pertussis patients have been studied: typical (group 1), atypical in their cultural properties (group 2), unidentified Gram-negative bacilli agglutinated by pertussis and parapertussis antitoxins (group 3). Besides, B. pertussis cultures, obtained by subculturing 2 museum strains and 2 newly isolated strains on synthetic casein-charcoal agar with subinhibiting doses of antibiotics or specific immune sera added, have been studied. As indicated by the results of this study, strains belonging to groups 1 and 2 contain glutamine synthetase, while in strains of group 3 this enzyme is absent. In immunoelectrophoresis strains of group 3 have been found to contain not a single antigen similar to the antigens of strains belonging to groups 1 and 2. Electrophoresis in polyacrylamide gel has revealed to differences in the protein spectrum of the strains of these 3 groups. The investigation has shown that the determination of glutamine synthetase and immunoelectrophoresis can be used for the differentiation of B. pertussis from similar Gram-negative bacilli. B. pertussis strains, changed as the result of experiments with antibiotics and specific immune sera, have also been shown to retain their antigenic composition and protein spectrum and to have no essential difference in the content of glutamine synthetase.     

Cytophysiological characteristics of Arabidopsis thaliana cultivated cells with disable perception of ethylene signal by the ETR1 receptor

Contradictory data about ethylene influence on cell growth and division prompted us to investigate cytophysiological characteristics of suspension cultures of Arabidopsis thaliana of wild type Col-0 and ert1-1 mutant carrying a point mutation in the site of ethylene binding by the ETR1 receptor. Some cytophysiological characteristics of the etr1-1 cultivated cells differed from those of Col-0: the growth rate of mutant cells was less and cell sizes were smaller, the culture was committed to the formation of tracheary elements (TE), had a pronounced modal class of nuclei (54%) with the amount of DNA 8C and a tendency to expand the ploidy toward 32C. Despite the absence of ethylene perception by the ETR1 receptor, the cell culture of mutant responded to treatment with ethylene by growth acceleration, an increase in cell viability and in the number of cells in the S-phase of the cell cycle. The inhibitor of ethylene binding to receptors, 1-methylcyclopropene, suppressed growth and viability of the cells of both genotypes. In the etr1-1 cell culture, the inhibitor reduced the number of S-phase nuclei and activated TE formation. All data obtained indicate that ethylene perception and transduction of ethylene signal are required for the maintenance of cell viability and active in vitro growth. It is supposed that the functional activity of the ETR1 receptor is necessary for optimal cell expansion, whereas other receptors are responsible for cell proliferation.     

The effect of NADP+ on cardiomyocyte electrophysiological properties of C57BL/6 and mdx mice

We have studied the influence of NADP⁺ on routine electrocardiography (ECG) in 6-month-old C57BL/6 and mdx mice. The animals were anesthetized by ether before ECG recording. ECG registration was carried out at a speed of 100 mm/s. The first ECG recording was made before intraperitoneal NADP⁺ injection in a dose of 13 or 80 mg/kg. The second ECG recording was made 10 min after NADP⁺ injection. Anesthesia was then terminated. The mice were occasionally anesthetized 45–60 min later, and the third ECG was recorded 1 h after injection of NADP⁺. ECG recording was carried out at a speed of 100 mm/s in standard leads I, II, and III and unipolar leads AvR, AvL, and AvF. Values of standard ECG characteristics, such as the P wave and the intervals PQ, QT, RR, and the QRS complex, were measured in milliseconds in standard lead II. We did not observe any differences between ECG magnitudes of 2- to 3-month-old C57BL/6 and mdx mice during trial experiments. Mice of both strains had a sinus rhythm in their heart rate. The QRS complex in mdx mice had a tendency to be larger than in C57BL/6 mice. Heart rates fluctuated between 722 ± 22 and 681 ± 21 beats per minute. The effect of NADP⁺ was studied in 6-month-old male mice. The increase in the RR interval and the decline in heart rate from 697 ± 21 to 461 ± 23 and 491 ± 28 beats per min for C57BL/6 mice (p < 0.01) and from 722 ± 28 beats per minute to 454 ± 31 beats per min for mdx mice were registered 10 min after NADP⁺ injection at a dose of 80 mg/kg. The increase in the RR interval can be explained by an increase in the QT interval. A statistically significant reduction in the QT interval leading to a diminished RR interval was observed in mdx mice 1 h after NADP⁺ injection. NADP⁺ at a dose of 13 mg/kg did not significantly change the ECG properties in mdx mice. ECG of mdx mice was characterized by negative repolarization of the T wave in 37% of all leads. The amount of leads with negative T-wave repolarization decreased up to 3% 1 h after NADP⁺ injection in dose of 80 mg/kg. The results have shown that cytomembranes of ventricular cardiac myocytes and the degree of oxidative stress are the main targets of the action of NADP⁺ in C57BL/6 and mdx mouse hearts.     

Spatiotemporal deep imaging of syncytium induced by the soybean cyst nematode <Emphasis Type="Italic">Heterodera glycines</Emphasis>

Parasite infections cause dramatic anatomical and ultrastructural changes in host plants. Cyst nematodes are parasites that invade host roots and induce a specific feeding structure called a syncytium. A syncytium is a large multinucleate cell formed by cell wall dissolution-mediated cell fusion. The soybean cyst nematode (SCN), Heterodera glycines, is a major soybean pathogen. To investigate SCN infection and the syncytium structure, we established an in planta deep imaging system using a clearing solution ClearSee and two-photon excitation microscopy (2PEM). Using this system, we found that several cells were incorporated into the syncytium; the nuclei increased in size and the cell wall openings began to be visible at 2 days after inoculation (DAI). Moreover, at 14 DAI, in the syncytium developed in the cortex, there were thickened concave cell wall pillars that resembled “Parthenon pillars.” In contrast, there were many thick board-like cell walls and rarely Parthenon pillars in the syncytium developed in the stele. We revealed that the syncytia were classified into two types based on the pattern of the cell wall structures, which appeared to be determined by the position of the syncytium inside roots. Our results provide new insights into the developmental process of syncytium induced by cyst nematode and a better understanding of the three-dimensional structure of the syncytium in host roots.     

Parallel double helix and tertiary structure of nucleic acids

The thermal denaturation of four oligonucleotides, viz. 3'-d(AT)5pO(CH2)6Opd(AT)5-3' (parAT), 3'-d(AT)5pO(CH2)5Opd(AT)5-5' (antiAT), 3'-d(A)10pO(CH2)6Op(T)10-3' (parA-T) and 3'd(A)10pOX X (CH2)6Opd(T)10-5' (antiA-T) in 0.01 M phosphate buffer at pH 7 in presence 0.1, 0.25, 0.5 and 1.0 M NaCl have been studied. It was shown that at lower temperature (0-20 degrees C) all oligomeres exist as complexes of two (canonic duplex) or four (eight) molecules of oligonucleotides, but at higher temperature (30-70 degrees C)- as hairpins with parallel (parAT and parA-T) of antiparallel (antiAT and antiA-T) orientation of chains. Thermodinamic parameters of separated strands-hairpins and hairpins--"low temperature complexes" transition were computated from the melting curves [A260 (T)] by nonlinear regression. AntiA-T was shown by ethidium bromide binding to exist at low strength (0.01 M phosphate buffer without NaCl) as four-stranded complex from two antiparallel double stranded helices parallely oriented and bonded by satisfy hydrogen-bond of groups not involved in WC-pairing. At higher ionic strength the two of such tetramers was conjugated by hydrophobic interaction into octamers. We speculate that four-stranded complexes serves to bring together, and zipper up two antiparallel double stranded helices at replication of DNA, cross-over of gomologues chromosomes and other biochemically important processes.     

Density and distribution of bacterioplankton and planktonic ciliates in the Bering Sea and North Pacific

The total number of planktonic bacteria in the upper mixed layerof the Bering Sea during the late spring-early summer periodranged between 1 and {small tilde}4 x 10⁶ ml^–1 (biomass10–40mg C m^–3). In the northern Pacific, along 47–52⁶N,the corresponding characteristics of the bacterioplankton densityin the upper mixed water layer were: total number 1–2x 10⁶ cells ml^–1 and biomass 15–46mg C m^–3Below the thermocline at 50–100 m, the density of bacterioplanktonrapidly decreased. At 300 m depth, it stabilized at 0.1–0.2x 106 cells ml^–1. The integrated biomass of bacterioplanktonin the open Bering Sea ranged between 1.2 and 3.6 g C m^–2(wet biomass 6–18 g m^–2) Its production per dayvaried from 2 to 23 mg C m^–3 days^–1 in the upper0–100 m. The numerical abundance of planktonic ciliatesin this layer was estimated to be from 3 to l0 x 10³ cells l^–1,and in the northern Pacific from 0.4 to 4.5 x 10³ l^–2.Their populations were dominated by naked forms of Strombidium,Strombilidium and Tontonia. In some shelf areas, up to 40% ofthe total ciliate population was represented by the symbioticciliate Mesodinium rubrum. The data on the integrated biomassof basic groups of planktonic microheterotrophs are also presented,and their importance in the trophic relationships in pelagiccommunities of subarctic seas is discussed.     

Early plant growth and biochemical responses induced by <Emphasis Type="Italic">Azospirillum brasilense</Emphasis> Sp245 lipopolysaccharides in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) seedlings are attenuated by procyanidin B2

This study analyzes the effects of procyanidin B2 on early wheat plant growth and plant biochemical responses promoted by lipopolysaccharides (LPS) derived from the rhizobacteria Azospirillum brasilense Sp245. Measurements of leaf, root length, fresh weight, and dry weight showed in vitro plant growth stimulation 4 days after treatment with A. brasilense as well as LPS. Superoxide anion (O₂ ^·?) and hydrogen peroxide (H₂O₂) levels increased in seedling roots treated with LPS (100 μg mL^?1). The chlorophyll content in leaf decreased while the starch content increased 24 h after treatment in seedling roots. The LPS treatment induced a high increase in total peroxidase (POX) (EC 1.11.1.7) activity and ionically bound cell wall POX content in roots, when compared to respective controls. Early plant growth and biochemical responses observed in wheat seedlings treated with LPS were inhibited by the addition of procyanidin B2 (5 μg mL^?1), a B type proanthocyanidin (PAC), plant-derived polyphenolic compound with binding properties of LPS. All results suggest first that the ionically bound cell wall POX enzymes could be a molecular target of A. brasilense LPS, and second that the recognition or association of LPS by plant cells is required to activate plant responses. This last event could play a critical role during plant growth regulation by A. brasilense LPS.