Bacillus anthracis but not always anthrax

Gram-positive bacilli isolated during epidemiological investigations which, on the basis of conventional tests, resemble Bacillus anthracis but which fail to produce the capsule or to induce anthrax in test animals have long been dismissed in clinical and veterinary laboratories as B. cereus or simply as unidentified Bacillus spp. and thereupon discarded as inconsequential. In this study, the application of newly available DNA probe, polymerase chain reaction and specific toxin antigen detection technology has revealed that a proportion of such strains are B. anthracis which lack the plasmid carrying the capsule gene (pXO2). While these techniques cannot, of course, be used to confirm the identities of strains resembling B. anthracis but which also lack the plasmid carrying the toxin genes (pXO1), the likelihood that these also are bonajide B. anthracis becomes more acceptable. (As yet no naturally occurring pXOl^-/2⁺ strains have been found.) At this point, the significance of the presence of such avirulent forms of B. anthracis in specimens can only be a subject for speculation, but the possibility that they may be indicators of virulent parents somewhere in the system being examined must be considered.     

Analysis of multi-type recurrent events in longitudinal studies; application to a skin cancer prevention trial

We consider the statistical modeling and analysis of replicated multi-type point process data with covariates. Such data arise when heterogeneous subjects experience repeated events or failures which may be of several distinct types. The underlying processes are modeled as nonhomogeneous mixed Poisson processes with random (subject) and fixed (covariate) effects. The method of maximum likelihood is used to obtain estimates and standard errors of the failure rate parameters and regression coefficients. Score tests and likelihood ratio statistics are used for covariate selection. A graphical test of goodness of fit of the selected model is based on generalized residuals. Measures for determining the influence of an individual observation on the estimated regression coefficients and on the score test statistic are developed. An application is described to a large ongoing randomized controlled clinical trial for the efficacy of nutritional supplements of selenium for the prevention of two types of skin cancer.     

Expression and secretion in Aspergillus nidulans and Aspergillus niger of a cell surface glycoprotein from the cattle tick, Boophilus microplus, by using the fungal amdS promoter system.

A cell surface glycoprotein (Bm86) from cells of the digestive tract of the cattle tick Boophilus microplus, which has been shown to elicit a protective immunological response in vaccinated cattle, was expressed and secreted in the filamentous fungi Aspergillus nidulans and Aspergillus niger by using the fungal amdS promoter system. The cloned gene coded for the Bm86 secretory signal and all of the Bm86 mature polypeptide except for the hydrophobic carboxy-terminal segment. High levels of Bm86 mRNA were detected in the transformed cells. Bm86 polypeptide was secreted from the cells in a soluble form and it was glycosylated, probably to a similar extent to the native glycoprotein. The recombinant product had an apparent molecular mass of 83 to 87 kilodaltons, whereas that predicted from the amino acid sequence was 69 kilodaltons. The Bm86 was expressed at levels of up to 1.8 mg/liter, or approximately 6% of secreted protein under the growth conditions used. No intracellular Bm86 was detected. A general relationship was observed between transformants containing a high number of copies of the expression plasmid and high expression levels.     

Tetracyano-2,2-bipyridineiron(iii), an improved electron acceptor for the spectrophotometric assay of beta-oxidation and of succinate dehydrogenase in intact mitochondria.

A recently described direct reading assay for beta-oxidation and for succinate oxidation in intact mitochondria using [Fe(CN)6]3- as final electron acceptor [Osmundsen & Bremer (1977) Biochem. J. 164. 621--633] has been improved by using instead tetracyano-2,2-bipyridineiron(III) [Fe(CN)4(bpy)]-, which gives a 2.6 times greater absorbance change on reduction. Some physical and kinetic properties of [Fe(CN)4(bpy)]- are described. The use of exogenous cytochrome c(III) as electron acceptor was also tested; this gives the largest absorbance change, although the absolute rate of reaction is only approx. one-third of that using [Fe(CN)6]3- or [Fe(CN)4(bpy)]-.     

Specific changes in the production of prostanoids by the ovine cervix at parturition

A method of tissue superfusion has been used to measure prostanoid production by the ovine cervix during late pregnancy and at parturition. In late pregnancy (105–135 days of gestation) cervical tissue produced relatively large amounts of prostaglandin E (PGE); in comparison, the production rates of prostaglandin F (PGF), 13, 14-dihydro-15-oxo-prostaglandin F (PGFM) and 6-oxo-prostaglandin F_1α were generally low. Thromboxane B₂ (TXB₂) production was minimal and often unmeasurable. There were significant increases in the production rates of PGE and 6-oxo-PGF_1α by cervical tissue taken immediately after delivery, when compared to late pregnancy. Mean production rates of PGE increased from 19.8 ± 4.1 to 43.8 ± 7.4 ng/g. dry wt./min; 6-oxo-PGF_1α production rates increased more than three-fold from 10.0 ± 2.7 to 34.6 ± 9.8 ng/g. dry wt./min (means ± S.E.M.). There were no significant differences in the rates of production of PGF, PGFM and TXB₂ by the two groups.     

The relation between patterns of ovarian follicle growth and ovulation rate in sheep

The number and growth rate of follicles within classes based on granulosa volume were determined for ovaries taken from groups of 4-5-year-old, fine-wool Merino ewes drawn at different times of the year from a single strain flock maintained at Armidale, N.S.W. The breeding season of the flock normally extends from February to October and the mean ovulation rate rises from about 0.5 in February to about 1.8-1.9 during April-May. Ewes sampled when they were anoestrous or had one (single-ovulatory) or two (twin-ovulatory) recent corpora lutea did not differ in respect to the mean total number of ovarian follicles, the mean number of follicles in individual classes, the time for follicles to complete their rapid growth stage, or the incidence of follicle atresia. However, the ovaries of twin-ovulatory ewes contained significantly more follicles in the two terminal classes within the rapid growth stage than did the ovaries of single-ovulatory or anoestrous ewes (2.2 v. 0.9 and 1.0). This difference was attributed to the differing numbers of follicles per day entering into the rapid growth stage (5.2, 4.5 and 3.7 respectively in twin-ovulatory, single-ovulatory and anoestrous ewes).     

Contribution of preterm delivery to perinatal mortality.

A detailed retrospective analysis was made of the records of 486 preterm infants, who accounted for 5-1% of all births during 1973 and 1974. Whereas preterm delivery did not contribute to perinatal mortality in terms of stillbirth, it outweighed all other causes in terms of early neonatal deaths. Preterm birth was responsible for 85% of the early neonatal deaths not due to lethal congenital deformities. Early neonatal mortality rates were closely linked both to gestational age and birth weight and to the reason for preterm birth. Early neonatal mortality was high (97 per 1000) when preterm labour was spontaneous, whether or not associated with material or fetal disease or with multiple pregnancy, but low (27 per 1000) when preterm delivery was elective. Preventing spontaneous preterm labour would considerably reduce neonatal mortality in our community.     

Sulfur mustards induce neurite extension and acetylcholinesterase synthesis in cultured neuroblastoma cells

When neuroblastoma cells (N18) in vitro were exposed to the bifunctional alkylating agent di-2-chloroethyl sulfide (HS), the specific activity of acetylcholinesterase began to rise rapidly after an initial lag period of 1 to 2 days. The five-fold increase in enzyme activity at 4 days after exposure to 0.5 μg/ml of HS was accompanied by a 25-fold rise in the rate of reappearance of acetylcholinesterase activity following essentially irreversible inhibition. Based on previous experience with acetylcholinesterase synthesis in serum deprived neuroblastoma cells, this behavior indicates induction of the enzyme. Vinblastine blocked the concomitant large increase in neurite extension which was stimulated by HS, but left acetylcholinesterase induction unaffected. Since enzyme activity was inversely related to the ability of the monolayer cells to form microcolonies, we conclude that acetylcholinesterase induction is dependent upon inhibition of cell division and independent of neurite extension. The monofunctional analogue of HS, 2-chloroethyl ethyl sulfide (CEES), produced similar effects, but much higher concentrations were required.     

Enzymes of glycollate formation and oxidation in two members of the rhodospirillaceae (Purple non-sulphur bacteria)

1. Phototrophic cultures of Rhodomicrobium vannielii do not excrete glycollate when gassed anaerobically with nitrogen plus carbon dioxide, although the addition of α-hydroxy-2-pyridine methanesulphonate (HPMS) results in the excretion of a trace amount of glycollate. The inclusion of low amounts of oxygen in this gas mixture results in marked glycollate excretion, higher rates occurring in the presence of HPMS.
2. Cell extracts of Rhodomicrobium vannielii, and also of Rhodospirillum rubrum, which excretes glycollate only under aerobic conditions in the light, catalyze the formation of glycollate from phosphoglycollate and also the oxidation of glycollate to glyoxylate.