Two fractions of N-Acetylmuramyl-l-alanine Amidase with Different Functions Isolated from Bacillus subtilis 168 Cells

N-acetylmuramyl-l-alanine amidase was obtained from exponentially growing Bacillus subtilis cells. The preparation contained two fractions whose molecular weights were 110,000 and 220,000. The two fractions had the same amidase activity and equally accelerated cell wall turnover of autolytic enzyme-deficient mutant cells. But the fraction with the lower molecular weight was more effective in dechaining the cell-chains of mutant cells.     

Metabolism of Isoxathion,O, O-Diethyl O-(5-Phenyl-3-isoxazolyl) phosphorothioate in the Rats

Excretion, distribution and metabolism of the insecticide, Isoxathion, administered orally in male Wistar-strain rats, were investigated with a carbon-14 labeled chemical. During 96 hr, approximately 85% and 14% of the total radioactivity were excreted in the urine and feces. Distribution of isoxathion after oral administration in the rats was investigated by means of whole-body autoradiographic technique and measurement of radioactivity in the tissues. At least eleven radioactive metabolites were detected, four of which were structurally determined. They were 3-hydroxy-5-phenylisoxazole, 3-(β-d-glucopyranuronosyloxy)-5-phenylisoxazole, 5-phenyl-3-isoxazolyl sulfate and hippuric acid.     

Fatty Acids as Inhibitors of Microbial Cell Wall Synthesis

The Maillard reaction of DNA with ketoses was investigated. Several days of incubation of d-fructose 6-phosphate with deoxyribonucleotides or with polymer DNA in an aqueous buffer resulted in the formation of chromophores and fluorophores. Aminoguanidine and sodium cvanoborohydride inhibited the formation of fluorophores. Transition metal ions such as Cu²⁺, Fe³⁺, Fe²⁺, or Mn^{2 +} promoted the formation of chromophores and fluorophores. Metal-chelating agents such as DETAPAC, citrate, and Desferal inhibited the formation of fluorophores. Superoxide dismutase and catalase also inhibited the formation of fluorophores. The transition metal ion-catalyzed autoxidation of d-fructose 6-phosphate or of the Heyns rearrangement products were to be partially involved in the glycation of DNA and subsequent formation of chromophores and of fluorophores.     

Syntheses of α and γ-BHC Alkylthio Analogs

Meso-(1245/36)-1,2,4,5,6-pentachloro-3-methylthiocyclohexane, and (124/356)-1,2,4,5,6-pentachloro-3-methylthio and ethylthiocyclohexanes were prepared from (1234/56)-1,4,5,6-tetrachloro-2,3-epoxycyclohexane (α-BTC cis-epoxide).     

Metabolism of Hymexazol, 3-Hydroxy-5-methylisoxazole,in the Rats

Excretion, distribution and metabolism of the fungicide, hymexazol, (3-hydroxy-5-methylisoxazole), labeled with carbon-14 were examined after administration of a single oral dose to Wistar-strain rats. Hymexazol was rapidly absorbed and distributed in the tissues. During 96 hr, 97% of the total radioactivity was excreted in the urine and 0.89% in the feces, and 0.86% was found in the expired gasses for 24 hr. Two metabolites were detected in the urine, whose chemical structures were determined as 3-(β-d-glucopyranuronosyloxy)-5- methylisoxazole and 5-methyl-3-isoxazolyl sulfate.     

Demographic differences in diet breadth of Canada lynx during a fluctuation in prey availability

Population dynamics of specialist carnivores are closely linked to prey availability, but the extent of variability in diet breadth of individual carnivores relative to natural variability in the abundance of their primary prey is not well understood. Canada lynx (Lynx canadensis) specialize on snowshoe hares (Lepus americanus) and exhibit cyclic fluctuations in abundance that lag 1–2 years behind those of snowshoe hares. Declining hare densities spur demographic changes in lynx, but it is unclear whether a corresponding increase in diet breadth occurs: (1) broadly across a lynx population; (2) only among individuals who are able to effectively switch to alternative prey; or (3) only among individuals who cannot capture sufficient primary prey. We measured stable isotope ratios of lynx muscle tissue spanning a cyclic increase and decline in hare density (1998–2001) in Fort Providence, NT, Canada. We found that lynx cohorts responded differently to hare population change, with yearling animals having broader diets at low hare densities, while adults and dependent juveniles maintained a constant diet through the initial decline in hare density. This result was consistent irrespective of lynx sex and indicates that yearling lynx likely are forced to adopt a broader diet when primary prey densities decline. Our results imply that select cohorts of specialist carnivores can exhibit high dietary plasticity in response to changes in primary prey abundance, prompting the need to determine whether increased diet breadth in young lynx is a successful strategy for surviving through periods of snowshoe hare scarcity. In this way, cohort‐specific niche expansion could strongly affect the dynamics of organisms exhibiting population cycles.     

An enzyme combination assay for serum sphingomyelin: Improved specificity through avoiding the interference with lysophosphatidylcholine

Serum sphingomyelin (SM) has predictive value in the development of atherosclerosis. Furthermore, SM plays important roles in cell membrane structure, signal transduction pathways, and lipid raft formation. A convenient enzymatic method for SM is available for routine laboratory practice, but the enzyme specificity is not sufficient because of nonspecific reactions with lysophosphatidylcholine (LPC). Based on the differential specificity of selected enzymes toward choline-containing phospholipids, a two-step assay for measuring SM was constructed and its performance was evaluated using sera from healthy individuals on a Hitachi 7170 autoanalyzer. Results from this assay were highly correlated with theoretical serum SM concentrations estimated by subtracting phosphatidylcholine (PC) and LPC concentrations from that of total phospholipids determined using previously established methods. There was a good correlation between the results of SM assayed by the proposed method and the existing enzymatic method in sera from healthy individuals. Moreover, the proposed method was superior to the existing method in preventing nonspecific reactions with LPC present in sera. The proposed method does not require any pretreatment, uses 2.5 μl of serum samples, and requires only 10 min on an autoanalyzer. This high-throughput method can measure serum SM with sufficient specificity for clinical purposes and is applicable in routine laboratory practice.     

Prion infection correlates with hypersensitivity of P2X7 nucleotide receptor in a mouse microglial cell line

We recently established mouse microglial cells persistently infected with mouse-adapted scrapie ME7 (ScMG20/ME7) for in vitro study of prion pathogenesis. Here, we found that ScMG20/ME7 cells were hypersensitive to P2X7 receptor agonists, as demonstrated by sustained Ca(2+) influx, membrane pore formation, cell death, and interleukin-1beta release. P2X7 mRNA expression was upregulated in these cells, and also in scrapie-infected mice brains. Treatment with pentosan polysulfate eliminated the infectivity and disease-related forms of prion protein from ScMG20/ME7 cell cultures, however, hypersensitivity of P2X7 receptors remained. These results suggest that prion infections may strongly affect the P2X7 receptor system in mouse microglial cells.     

Abeta42 overproduction associated with structural changes in the catalytic pore of gamma-secretase: common effects of Pen-2 N-terminal elongation and fenofibrate

gamma-Secretase is an atypical aspartyl protease that cleaves amyloid beta-precursor protein to generate Abeta peptides that are causative for Alzheimer disease. gamma-Secretase is a multimeric membrane protein complex composed of presenilin (PS), nicastrin, Aph-1, and Pen-2. Pen-2 directly binds to transmembrane domain 4 of PS and confers proteolytic activity on gamma-secretase, although the mechanism of activation and its role in catalysis remain unknown. Here we show that an addition of amino acid residues to the N terminus of Pen-2 specifically increases the generation of Abeta42, the longer and more aggregable species of Abeta. The effect of the N-terminal elongation of Pen-2 on Abeta42 generation was independent of the amino acid sequences, the expression system and the presenilin species. In vitro gamma-secretase assay revealed that Pen-2 directly affects the Abeta42-generating activity of gamma-secretase. The elongation of Pen-2 N terminus caused a reduction in the water accessibility of the luminal side of the catalytic pore of PS1 in a similar manner to that caused by an Abeta42-raising gamma-secretase modulator, fenofibrate, as determined by substituted cysteine accessibility method. These data suggest a unique mechanism of Abeta42 overproduction associated with structural changes in the catalytic pore of presenilins caused commonly by the N-terminal elongation of Pen-2 and fenofibrate.