Structural characterization of lipid A component of Erwinia carotovora lipopolysaccharide

The chemical structure of the lipid A component of lipopolysaccharide excreted into the liquid medium by the plant pathogenic enterobacterium Erwinia carotovora FERM P-7576 was characterized. It consists of a -1, 6-linked glucosamine disaccharide which carries ester-and amide-bound fatty acids and phosphate similar to the lipid A from other gram-negative bacteria. The lipid A preparation was not uniform in the number and composition of the fatty acids linked to the disaccharide. Four prominent lipids A were involved, they were composed of five to seven residues of fatty acid. Among them the major component was hexa-acyl lipid A, in which the hydroxyl group at position 3 and the amino group of the non-reducing glucosamine unit carry 3-dodecanoyl-oxytetradecanoyl residues. Positions 2 and 3 of the reducing glucosamine unit were substituted by 3-hydroxytetradecanoic acid. In the hepta-acyl lipid A, an additional hexadecanoic acid was linked to the hydroxyl group of the 3-hydroxytetradecanoyl residue at position 2 of the hexa-acyl lipid A. Two penta-acyl lipids A were the homologs of the hexa-acyl lipid A with decreasing acylation. Dodecanoic acid was missing from one, and 3-hydroxytetradecanoic acid from another. 3-Dodecanoyloxytetradecanoyl residue at position 3 differentiates E. carotovora lipid A from that of other gram-negative bacteria.Abbreviations LPS lipopolysaccharide - GlcN glucosamine - KDO 3-deoxy-d-manno-octulosonic acid - FAB-MS fast atom bombardment mass spectrometry - u atomic mass unit     

Supernumerary chromosomes in the human bed bug,Cimex lectularius Linn. (Cimicidae:Hemiptera)

The chromosome complement in the human bed bug, Cimex lectularius Linn., is 26+X₁X₂Y in the male and 26+X₁X₁X₂X₂ in the female. However, a population from Cairo, Egypt has 4 supernumerary X chromosomes. In the hybrid between the Berkeley population (with no supernumeraries) and the Cairo population (with 4 supernumeraries), the behavior of supernumeraries was observed during embryogenesis and oogenesis as well as spermatogenesis.In embryogenesis the transmission of supernumeraries was quite regular. However, one chromosome may sometimes be eliminated early in the germ line. This abnormality could induce the variations in chromosome number encountered in later stages. In the first meiotic division, some of the supernumeraries were nondisjunctional. Moreover, in the second division, some supernumeraries were eliminated. These results show that there is a tendency towards a decrease in the number of supernumeraries in the hybrids.Although the supernumeraries behave like X chromosomes, they seem not to be important for sex determination and appear to be largely or entirely inert genetically. Supernumeraries in the bed bug originate from small fragments caused by structural rearrangement. They are increased by an accumulation mechanism. Supernumeraries in the bed bug appear to be of relatively recent origin. The phylogenetic sequence in their development was probably from none to a stabilized number of four in the Old World. Then the supernumeraries were lost in two specialized lines, Cimex columbarius Jenyns on domestic birds in Europe and Cimex lectularius Linn. on man in the Western Hemisphere.This study was carried out under U.S. Public Health Service Grant (GM-13197).     

scRNA-seq Analysis of Hemocytes of Penaeid Shrimp Under Virus Infection

Marine Biotechnology - The classification of cells in non-model organisms has lagged behind the classification of cells in model organisms that have established cluster of differentiation marker...     

Evolution of Pulmonate Gastropod Mitochondrial Genomes: Comparisons of Gene Organizations of Euhadra, Cepaea and Albinaria and Implications of Unusual Trna Secondary Structures

Complete gene organizations of the mitochondrial genomes of three pulmonate gastropods, Euhadra herklotsi, Cepaea nemoralis and Albinaria coerulea, permit comparisons of their gene organizations. Euhadra and Cepaea are classified in the same superfamily, Helicoidea, yet they show several differences in the order of tRNA and protein coding genes. Albinaria is distantly related to the other two genera but shares the same gene order in one part of its mitochondrial genome with Euhadra and in another part with Cepaea. Despite their small size (14.1-14.5 kbp), these snail mtDNAs encode 13 protein genes, two rRNA genes and at least 22 tRNA genes. These genomes exhibit several unusual or unique features compared to other published metazoan mitochondrial genomes, including those of other molluscs. Several tRNAs predicted from the DNA sequences possess bizarre structures lacking either the T stem or the D stem, similar to the situation seen in nematode mt-tRNAs. The acceptor stems of many tRNAs show a considerable number of mismatched basepairs, indicating that the RNA editing process recently demonstrated in Euhadra is widespread in the pulmonate gastropods. Strong selection acting on mitochondrial genomes of these animals would have resulted in frequent occurrence of the mismatched basepairs in regions of overlapping genes.     

Kinetics of growth and lactic acid production in continuous and batch culture

Summary In a lactic acid fermentation by Streptococcus faecalis, the specific consumption rate of glucose (v) and the specific production rate of lactic acid () were represented by the following simple equations as functions of the specific growth rate (): 1/=(1/) + 1/ = (1/) + By use of data from a batch culture, these two equations were derived from enzyme kinetics of the product inhibition. These equations were successfully applied to the results of batch culture and chemostat culture. In addition, calculation of ATP yield by these equations agreed with the experimental results better than the conventional Leudeking-Piret type equation, which includes two terms associated with growth and not with growth. Correspondence to: H. Ohara     

Aldehyde reductase gene expression by lipid peroxidation end products,MDA and HNE

Membrane lipid peroxidation results in the production of a variety of aldehydic compounds that play a significant role in aging, drug toxicity and the pathogenesis of a number of human diseases, such as atherosclerosis and cancer. Increased lipid peroxidation and reduced antioxidant status may also contribute to the development of diabetic complications. This study reports that lipid peroxidation end products such as malondialdehyde (MDA) and 4-hydroxynonenal (HNE) induce aldehyde reductase (ALR) gene expression. MDA and HNE induce an increase in intracellular peroxide levels; N-Acetyl-L-cysteine (NAC) suppressed MDA- and HNE-induced ALR gene expression. These results indicate that increased levels of intracellular peroxides by MDA and HNE might be involved in the upregulation of ALR.     

Genetic exchange between two freshwater apple snails, Pomacea canaliculata and Pomacea maculata invading East and Southeast Asia

Two species of apple snails, Pomacea canaliculata and Pomacea maculata (formerly Pomacea insularum), have invaded many countries of East and Southeast Asia from their native range in South America. This study investigated the genetic structure of the two species invading these areas. Phylogenetic analysis based on sequences of the nuclear gene elongation factor 1-alpha (EF1α) detected two well-supported clades (Clade C and Clade M). Both P. canaliculata and P. maculata were represented in each clade. Some snails had both Clade C and Clade M EF1α sequences. These results suggest genetic exchange between snails of the two clades. A mating experiment between P. canaliculata with Clade C EF1α sequences and P. maculata with Clade M EF1α sequences resulted in viable F1 progeny under laboratory conditions. The genetic exchange was also inferred in some populations collected from Argentina, suggesting an existence of hybrid in the native range. Simple identification of EF1α types using a restriction enzyme, ApaLI, detected significant geographical structure of the EF1α variants in the invaded area. The divergent geographical structure could have resulted from either the founder effect or the bridgehead effect, although further genetic analysis is needed to clarify this. Average individual egg weight, which is an indicator of egg size, was higher in P. canaliculata than P. maculata in both field and laboratory reared samples, suggesting that some (probably most) P. canaliculata and P. maculata invading East and Southeast Asia still maintain species-specific populations.     

Development of Iron-Phosphate Biofilms on Pyritic Mine Waste Rock Surfaces Previously Treated with Natural Phosphate Rocks

Various treatments have been proposed to attenuate and eventually inhibit the generation of acid mine drainage (AMD) or acid-rock drainage (ARD). The addition of Natural Phosphate Rocks (NPR) to mining wastes has been shown to reduce acid generation. The biogeochemical reactions underlying the phosphate precipitation reactions are however poorly understood, even though the chemical reactions are well defined. The present study was designed to study the role of solution chemistry and bacterial activity on phosphate precipitation on waste rock surfaces. Waste rock samples (rich in sulphides) previously weathered for 989 days in the presence of NPR were submersed in 2 different phosphate-rich growth media in order to enhance the growth of acidophilic and neutrophilic bacteria. DAPI and FISH analyses revealed that most cells belonged to the bacteria domain, and that alpha- and beta-proteobacteria were the dominant neutrophiles. ESEM, SEM and TEM observations of the samples revealed the presence of a biofilm on the surface of the rocks at both pH conditions. Bacteria and fine-grained precipitates were trapped in an exopolymer matrix. At low pH, the formation of fine precipitates rich in Fe and P within the biofilm corresponded to a decline of phosphate concentrations in the growth medium. This was in agreement with the solubility calculations which indicated that the medium became over-saturated with respect to some Fe-phosphate minerals. In the pH neutral system, solubility calculations indicated that Ca- and Mg-phosphate minerals were stable, but they were not detected in the biofilm. Solubility calculations also indicated that vivianite became unstable over time, which could explain the release of soluble phosphate over time in the pH neutral system. Our results showed that precipitation reactions played an important role in the solubility of phosphate in both systems, but a series of complex nucleation reactions involving bacterial exopolymers and the presence of microenvironments within the biofilms were likely important factors as well. Our findings also imply that the reduction of acid generation in NPR-treated waste rocks could be due in part to the formation of biofilms on the rock surfaces because the biofilms would act as a physical and chemcial barrier to limit the extent of pyrite oxidation.     

Effect of Additional Protein or Methionine and Threonine on Tyrosine Catabolism in Rats Fed Diets High in Tyrosine

The supplementation of additional protein or methionine and threonine to a high tyrosinc-low protein diet has previously been shown to prevent the tyrosine toxicity. To elucidate the mechanism, studies were performed on the effect of these supplements on the capacity to oxidize excessive tyrosine. Male weanling rats were ad libitum fed a 10% casein diet containing 5% tyrosine with and without extra casein or methionine plus threonine for 7 days, then animals were injected intraperitoneally with l-tyrosine-U-¹⁴C and the oxidation rate to ¹⁴CO₂ determined in vivo at an interval of several hours throughout a 24-hour period. The addition of extra casein or methionine and threonine to the high tyrosine diet enhanced the ability of tyrosine oxidation, and decreased the radioactivities of the TCA-soluble fractions in plasma, liver and muscle. A high level of free tyrosine in blood and tissues was also lowered by the addition of these supplements. The diurnal chenges in free tyrosine concentration of various tissues were observed. The data suggest that the beneficial effect of extra casein or methionine and threonine supplementation on tyrosine toxicity is due to an increased rate of tyrosine catabolism which results in lower tyrosine concentrations in body fluids which overcomes tyrosine toxicity.