Analysis of ribosomal RNA intergenic spacer (IGS) sequences in species and populations of Gyrodactylus (Platyhelminthes: Monogenea) from salmonid fish in Northern Europe

The intergenic spacer (IGS) region of ribosomal RNA genes was amplified and sequenced from a variety of Gyrodactylus specimens collected from wild and farmed Atlantic salmon Salmo salar, rainbow trout Oncorhynchus mykiss, and grayling Thymallus thymallus, from various locations in Northern Europe. Phylogenetic analysis of the sequences confirmed the distinction between G. salaris Malmberg, 1957 and G. thymalli Zitnan, 1960, supporting their validity as separate species. G. salaris adapted to rainbow trout are also distinct from the parasites found on Atlantic salmon, supporting the existence of a rainbow-trout form that was initially identified on the basis of morphological differences. Analysis of the IGS did not provide good resolution of different populations of G. salaris sensu stricto, but was consistent with epidemiological evidence which indicates that introduction of the parasite to Norway was recent and limited. The IGS may be helpful in distinguishing forms of G. salaris that are pathogenic to Atlantic salmon from those that are not.     

Effect of gene amplification on mercuric ion reduction activity of Escherichia coli.

The mercury resistance (mer) operon of plasmid R100 was cloned onto various plasmid vectors to study the effect of mer gene amplification on the rate of Hg2+ reduction by Escherichia coli cells. The plasmids were maintained at copy numbers ranging from 3 to 140 copies per cell. The overall Hg2+ reduction rate of intact cells increased only 2.4-fold for the 47-fold gene amplification. In contrast, the rate of the cytoplasmic reduction reaction, measured in permeabilized cells, increased linearly with increasing gene copy number, resulting in a 6.8-fold overall amplification. RNA hybridizations indicated that mRNA of the cytoplasmic mercuric reductase (merA gene product) increased 11-fold with the 47-fold gene amplification, while mRNA of the transport protein (merT gene product) increased only 5.4-fold. Radiolabeled proteins produced in maxicells were used to correlate the expression levels of the mer polypeptides with the measured reduction rates. The results indicated that, with increasing gene copy number, there was an approximately 5-fold increase in the merA gene product compared with a 2.5-fold increase in the merT gene product. These data demonstrate a parallel increase of Hg2+ reduction activity and transport protein expression in intact cells with plasmids with different copy numbers. In contrast, the expression level of the mercuric reductase gene underwent higher amplification than that of the transport genes at both the RNA and protein levels as plasmid copy number increased.     

Molecular and morphological comparisons between Gyrodactylus ostendicus n. sp. (Monogenea: Gyrodactylidae) on Pomatoschistus microps (Krøyer) and G. harengi Malmberg, 1957 on Clupea harengus membras L.

Gyrodactylus ostendicus n. sp. was exclusively found on fins of the common goby Pomatoschistus microps (Krøyer). The haptoral hard parts are among the smallest described for species of Gyrodactylus. A presumed similarity between the new species and G. harengi Malmberg, 1957 (subgenus Metanephrotus Malmberg, 1964) encouraged a comparative approach. A morphological analysis showed the marginal hook sickles of G. ostendicus to be of quite a different type and similar to those of G. arcuatus Bychowsky sensu Bychowsky &; Poljansky (1953) (subgenus Mesonephrotus Malmberg, 1964). The new species has a pharynx with short pharyngeal processes. Its protonephridial system has small bladders, indicating an association with the subgenera Mesonephrotus or Metanephrotus. Molecular phylogenetic analyses, including all of the species of Mesonephrotus and Metanephrotus currently available on the GenBank database, suggested that the new species belongs to Mesonephrotus. Combined morphological and molecular studies of the new species show that G. ostendicus is more closely related to G. arcuatus than to G. harengi.     

Characterization of the photosynthetic apparatus in cortical bark chlorenchyma of Scots pine

Winter-induced inhibition of photosynthesis in Scots pine (Pinus sylvestris L.) needles is accompanied by a 65% reduction of the maximum photochemical efficiency of photosystem II (PSII), measured as F _v/F _m, but relatively stable photosystem I (PSI) activity. In contrast, the photochemical efficiency of PSII in bark chlorenchyma of Scots pine twigs was shown to be well preserved, while PSI capacity was severely decreased. Low-temperature (77 K) chlorophyll fluorescence measurements also revealed lower relative fluorescence intensity emitted from PSI in bark chlorenchyma compared to needles regardless of the growing season. Nondenaturating SDS-PAGE analysis of the chlorophyll–protein complexes also revealed much lower abundance of LHCI and the CPI band related to light harvesting and the core complex of PSI, respectively, in bark chlorenchyma. These changes were associated with a 38% reduction in the total amount of chlorophyll in the bark chlorenchyma relative to winter needles, but the Chl a/b ratio and carotenoid composition were similar in the two tissues. As distinct from winter pine needles exhibiting ATP/ADP ratio of 11.3, the total adenylate content in winter bark chlorenchyma was 2.5-fold higher and the estimated ATP/ADP ratio was 20.7. The photochemical efficiency of PSII in needles attached to the twig recovered significantly faster (28–30 h) then in detached needles. Fluorescence quenching analysis revealed a high reduction state of Q _A and the PQ-pool in the green bark tissue. The role of bark chlorenchyma and its photochemical performance during the recovery of photosynthesis from winter stress in Scots pine is discussed.     

Steroid cyclicsulfites. III. Conformational and configurational aspects of steroid 3,5-cyclicsulfites.

The reaction of 3beta, 5beta-dihydroxy cholestanes with thionyl chloride is shown to yield cyclicsulfite esters containing boat heterocyclic rings with the S=O oxygen axial or equatorial, depending upon the mode of formation. Treatment of a diol in pyridine at low temperature favors an equatorial S=O conformation while higher reaction temperatures in chloroform solution yield a mixture of axial and equatorial epimers. In the case of a 7alpha-bromo-6-oxo 3,5-sulfite, it has been shown that the S=O equatorial isomer may be converted to the axial isomer upon treatment with acid.     

Kinetic analysis of cephalosporin biosynthesis in Streptomyces clavuligerus

A kinetic model describing the cephalosporin biosynthesis in Streptomyces clavuligerus was developed. Using previously reported kinetic data of biosynthetic enzymes, we examined the kinetics of cephalosporin production. The predicted time profile of the specific production rate during a batch culture parallels that of experimental observation. Sensitivity analysis reveals that delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (ACV) synthetase is the rate-limiting enzyme. The effect of amplifying ACV synthetase on the specific production rate was analyzed theoretically. Increasing ACV synthetase enhances the production rate initially until ACV synthetase enhances the production rate initially until deacetocycephalosporin C hydroxylase becomes rate-limiting. Such kinetic analysis can provide a rational basis for modifying the biosynthetic machinery of cephalosporin through gene cloning.     

Localization of lipids in the aortic wall with imaging TOF-SIMS

Time-of-flight secondary-ion-mass-spectrometry (TOF-SIMS) was utilized to address the issue of localization of lipids and inorganic ions in healthy rat aorta and human atherosclerotic plaque. Pieces of rat aorta were high pressure frozen, freeze-fractured and freeze dried. The samples were analyzed by imaging TOF-SIMS equipped with a Bi(1-7)(+)-source. Reference lipid samples were analyzed and compared to data obtained by analysis of the rat aorta samples. Fatty acids, cholesterol, oxysterol and diacylglycerols were detected and localized. A heterogeneous lipid distribution could be shown in the aorta, where the lamellae of the aorta, distinguished by imaging of CN(-), appeared enriched in cholesterol, oxysterol and diacylglycerols, while the smooth muscle tissue, identified by imaging of PO(3), appeared enriched in phosphocholine. Palmitic/palmitoleic acid and stearic/oleic acid appeared to be heterogeneously distributed over the aorta with high concentration areas located especially in the tunica media region of the aorta. Human atherosclerotic plaque showed an irregular cholesterol distribution mainly located in spots in the intima region with elongated diacylglycerol regions located mainly in the media region.