Comparison of methods used in zooplankton sampling and counting in the joint Russian-Finnish evaluation of the trophic state of Lake Ladoga

The results of a joint Russian-Finnish investigation on zooplankton in Lake Ladoga are presented, and a comparison is made between two sampling techniques, tube sampler and plankton net, and between two counting methods. The precision of subsampling and sampling is further discussed on the basis of zooplankton data gathered in Lake Saimaa, Finland. The comparison clearly indicates that a tube sampler is required for reliable sampling of small-sized animals, while a plankton net saves time and is a more economical sampler for large, rare or active animals. The comparison between the results obtained by Finnish and Russian workers, using different counting procedures, shows that the main groups of crustacean zooplankton are similarly counted and identified in the two laboratories.     

COORDINATED CAROTENOID AND LIPID SYNTHESES INDUCED IN PARIETOCHLORIS INCISA (CHLOROPHYTA,TREBOUXIOPHYCEAE) MUTANT DEFICIENT IN Δ5 DESATURASE BY NITROGEN STARVATION AND HIGH LIGHT1

The responses to PAR intensity and nitrogen deficiency have been investigated in the Δ5‐desaturase‐deficient mutant (P127) of the microalga Parietochloris incisa (Reisigl) Shin Watan. (Chlorophyta, Trebouxiophyceae). The mutant accumulates dihomo‐γ‐linolenic acid (DGLA, C20:3 ω6) instead of arachidonic acid (C20:4 ω6) characteristic of the wildtype. The growth, fatty acid and pigment composition, and light absorption by P127 cell suspensions were studied for the first time during cultivation on complete and N‐free BG‐11 medium at 35, 130, and 270 μE · m^?2 · s^?1. On complete medium under high irradiance, an increase in biomass was observed, and total fatty acid (TFA) and DGLA contents were higher than in N‐starving cultures. A distinct irradiance‐dependent rise in carotenoid‐to‐chl ratio was recorded in P127 due to an increase in carotenoids (on complete medium) or by a decline in chl (on N‐free medium). Cultivation under high and medium irradiances caused a decline in light‐harvesting xanthophylls and an increase in β‐carotene, localized predominantly in cytoplasmic oil bodies (OB). The P127 mutant, similar to wildtype, responded to the stresses by coordinated induction of fatty acid and carotenoid syntheses, but displayed the same magnitude of the response as was observed in wildtype under 30% lower irradiance. The changes in optical properties of the P127 cultures tightly correlated with their pigment composition, and hence with fatty acid content, making it possible to develop a nondestructive technique for the assay of TFA and DGLA. The peculiarities of the stress responses in the wildtype and the mutant are discussed.     

Construction of the new <Emphasis Type="Italic">Escherichia coli</Emphasis> K-12 MG 1655 novel strain with improved growth characteristics for application in metabolic engineering

MG1655 of Escherichia coli K-12 is frequently used in metabolic engineering as the wild-type strain. However, its two mutations, ilvG and rph-1 provide a negative effect on culture growth. The “polar effect” of rph-1 decreases the level of pyrE expression, causing partial auxotrophy for pyrimidines. Mutation ilvG leading to the appearance of Val^S phenotype causes retardation of cell growth rate on media containing amino acids. In this work, the substitution of two loci in the genome of MG1655 with the recovery of the wild-type phenotype was accomplished. Gene rph ^wt from the chromosome of E. coli TG1 was marked via Red-dependent integration of DNA fragment carrying λattL-Cm^R-λattR and transduced with phage P1 into MG1655; later, the Cm^R marker was removed with the use of λXis/Int recombinase. Parallel to this procedure, a spontaneous Val^R mutant of E. coli MG1655 yielding colonies of maximal size on M9 medium with glucose in the presence of L-Val (50 μg/ml) was isolated. It was shown that a nucleotide deletion in the isolated Val^R strain had been generated in the region of the identified ilvG mutation, which led to the recovery of the reading frame and active protein synthesis. This mutation named ilvG-15, which is the only reason for the Val^R phenotype in the obtained strain, was transferred to MG1655-rph ^wt using cotransduction, by analogy to the transfer of rph ^wt. Evaluation of rates of aerobically growing cells (μ, hour-1) on M9 medium with glucose produced the following values: 0.56, 0.69, and 0.73 for strains MG1655,MG1655-rph ^wt, and MG1655-(rph ^wt, ilvG-15), respectively.     

Ozone-induced oxidative modification of plasma fibrin-stabilizing factor

The plasma fibrin-stabilizing factor (pFXIII) function is to maintain a hemostasis by the fibrin clot stabilization. The conversion of pFXIII to the active form of the enzyme (FXIIIа) is a multistage process. Ozone-induced oxidation of pFXIII has been investigated at different stages of its enzyme activation. The biochemical results point to a decrease of an enzymatic activity of FXIIIа depending largely on the stage of the pFXIII conversion into FXIIIа at which oxidation was carried out. UV-, FTIR- and Raman spectroscopy demonstrated that chemical transformation of cyclic, NH, SH and S–S groups mainly determines the oxidation of amino acid residues of pFXIII polypeptide chains. Conversion of pFXIII to FXIIIa proved to increase protein sensitivity to oxidation in the order: pFXIII < pFXIII activated by thrombin < pFXIII in the presence of calcium ions < FXIIIa. The dynamic light scattering data indicate that the three-dimensional structure of pFXIII becomes loosened due to oxidative modification. ESR spectroscopy data also point to conformational changes of the fibrin-stabilizing factor under oxidation. Taking into account these new findings it seems reasonable to assume that the inhibitory/carrier FXIII-B subunits can serve as scavengers of ROS. Hypothetically, this mechanism could help to protect the key amino acid residues of the FXIII-A subunits responsible for the enzymatic function of FXIIIa.