Critical considerations in the development of an assay for sulfidopeptide leukotrienes in plasma

A sensitive and specific assay has been developed for measurement of total sulfidopeptide leukotrienes (LT) in plasma. LTC4 and LTD4 in plasma are converted to LTE4 which is then extracted by C18 Sep-Pak binding and elution. Total LTE4 is resolved by reverse phase high performance liquid chromatography (RP-HPLC) and quantitated by radioimmunoassay (RIA). A [3H]LTE4 internal standard is added to the starting plasma sample to allow overall recovery to be calculated and to define the fractions from RP-HPLC to be assayed for LTE4-like immunoreactivity. The correlation between the measured increase in LTE4 concentration after addition of incremental amounts of LTC4 and LTE4 to plasma was 0.989 and 0.978, respectively, with slopes of 1.05 and 1.11. Addition of 51 pg/ml LTE4 to 5 ml plasma was detectable; the measured increase was 48 +/- 12 pg/ml (mean +/- SE, n = 7). The intra-assay coefficient of variation for 341 pg/ml of added LTC4 was 3.2% (n = 6). Sulfidopeptide leukotrienes could not be detected in blood samples taken from 12 normal volunteers in whom the theoretical detection limit, calculated from the sensitivity of the RIA, the overall recovery of LTE4, and the volume of plasma extracted, was 83 +/- 4 pg LTE4/ml plasma (0.19 +/- 0.01 pmol sulfidopeptide leukotriene/ml plasma; mean +/- SE).     

Attachment as a factor in the protection of Enterobacter cloacae from chlorination

Enterobacter cloacae attached to drinking water distribution particles was subjected to chlorination. Attachment resulted in the protection of these organisms from disinfection. This effect was found to be dependent upon both the level of chlorine in the system and attachment time. The results obtained in this study indicate that attached organisms may play an important role in coliform outbreaks.     

Purification of the parasporal crystals ofBacillus thuringiensis subsp.israelensis and development of a novel bioassay technique

Summary A spore-free parasporal crystal suspension was prepared fromBacillus thuringiensis subsp.israelensis by an aqueous biphasic separation technique and the waste fractions quantified with regard to spore numbers and insecticidal potencies. The technique proved efficient in selectively removing spores from the spore-crystal mixture. The final crystal suspension was used to develop a novel bioassay system which allowed rapid determination of crystal effectiveness in 1/8 to 1/6 of the time required in the conventional mortality versus concentration bioassay.     

Microzooplankton and macrozooplankton glutamate dehydrogenase as indices of the relative contribution of these fractions to ammonium regeneration in the Gulf of Maine

Ammonium regeneration by micro- (35–153 µm) andmacrozooplankton (> 153 µm) was determined in the Gulfof Maine by measuring the activity of the excretory enzyme glutamatedehydrogenase (GDH) in various size fractions. GDH maxima weregenerally observed to correspond to the depth of the chlorophyllmaximum as previously reported in the Gulf of Mexico and inthe vicinity of the Nantucket Shoals. GDH activity of the microzooplanktonwas considerably lower than the macrazooplankton, suggestingthe microzooplankton made only a minor contribution (1–11%) to the total ammonium regenerated. These results were confirmedby biomass estimates made from counts of individual species.Ammonium excretion by both zooplankton fractions was estimatedto supply 5–31 % of the nitrogen requirements for primaryproduction, with an estimated 59–63% supplied by the verticaltransport of nitrate (new nitrogen) into the euphoric zone.     

Purification of bovine liver histidyl-tRNA synthetase, the Jo-1 antigen of polymyositis: size of the whole enzyme and its characteristic proteolytic fragments

We have recently reported the marked increase in frequency which can be achieved in the detection of the anti-Jo-1 antibody of polymyositis in serum samples by replacing commercial mixtures of cytoplasmic and nuclear antigens with the purified antigen, histidyl-tRNA synthetase. The present paper describes a method for purifying this antigen and an investigation of its size. Molecular masses previously reported for the enzyme have varied from 85-154 kDa and subunit molecular masses varying from 40-77 kDa have been observed. Several of these fragments are of sizes similar to those of a number of other autoantigens commonly observed in connective tissue diseases. Since the clinical identification of these autoantigens often relies exclusively on size determination by Western blotting, we have characterized the commonly occurring fragments of histidyl-tRNA synthetase lest they confuse such identification. It is concluded that histidyl-tRNA synthetase, like many other aminoacyl-tRNA synthetases, is subject to severe proteolysis during extraction procedures. Several characteristic fragments (Mr = 80, 75, 61, 55, 50 and 45 kDa) result, a finding that provides a satisfactory explanation of the various values previously reported. The intact bovine enzyme is a dimer of molecular mass close to 160 kDa.     

The association between spontaneous pyelonephritis and maturity-onset diabetes mellitus in male MM mice

Blood glucose and glucose tolerance tests demonstrated that many male MM mice are diabetic. Serial urine sampling showed that the diabetes occurred only in mature MM males and consisted of a single self-limiting episode. Histological examination of the pancreas, together with measurements of body weight, glycosylated haemoglobin and plasma insulin, revealed that the diabetes was of the maturity-onset insulin-resistant type. Bacteriological examination of the urine samples showed that urinary tract infection, a known feature of male MM mice, occurred in the diabetics but only after the onset of hyperglucosuria. It was concluded that the high urinary glucose levels of diabetic MM males are of prime importance in the aetiology of the renal infection which occurs rarely in non-diabetic MM males or in other strains in the colony. An infectious aetiology for the diabetes per se was excluded by the existence of diabetes in germfree MM males.     

Antitrypanosomal action of cis-diamminedichloroplatinum (II) analogs

The present report deals with the alterations produced by cis-diamminedichloroplatinum (II) (DDP), and 2 of its analogs: cis-Pt(II)(tranylcypromine)2Cl2 and cis-Pt(II)(benzothiazole)2Cl2 in cultured epimastigote forms of Trypanosoma cruzi. Studies have been performed at the ultrastructural level and the inhibitory effect of these complexes on macromolecule synthesis, evaluated by 3H-thymidine, 3H-uridine, and 3H-leucine incorporation, has been investigated. DDP at concentrations of 50 and 100 micrograms/ml does not inhibit significantly the incorporation of radioactive precursors, but a clear decrease was observed with the 2 analogs. Eight hours of treatment at a concentration of 10 micrograms/ml rendered in all 3 cases an increase in autophagic vacuoles and lipids as well as an abnormal condensation of the nucleus chromatin.     

The number of species of rodent coccidia and of other protozoa

About 447 species of coccidia have been named from the 1687 living, known species of rodents; 207 host species, 92 host genera, and 15 host families are represented; this is about 12% of the known species of rodents. About 4600 species of apicomplexan protozoa have been named. Assuming that the same proportion of the total number of apicomplexan species has been named as of the coccidian species, there must actually be about 38,333 species of apicomplexan protozoa. There are 5.4 times as many protozoan genera as of apicomplexan genera. Assuming that the number of species in each genus is the same for all the protozoa as it is for the Apicomplexa, there may actually be 206,998 species of protozoa. This may be too conservative an estimate. Based on other criteria, an estimate of over 20 million species could be made.