The use of 6Li{7Li}-REDOR NMR spectroscopy to compare the ionic conductivities of solid-state lithium ion electrolytes.

Garnet-like solid-state electrolyte materials for lithium ion batteries are promising replacements for the currently-used liquid electrolytes. This work compares the temperature dependent Li(+) ion hopping rate in Li6BaLa2M2O12 (M = Ta, Nb) using solid-state (6)Li{(7)Li}-REDOR NMR. The slope of the (6)Li{(7)Li}-REDOR curve is highly temperature dependent in these two phases, and a comparison of the changes of the REDOR slopes as a function of temperature has been used to evaluate the Li(+) ion dynamics. Our results indicate that the Nb phase has a higher overall ionic conductivity in the range of 247 K to 350 K, as well as a higher activation energy for lithium ion hopping than the Ta counterpart. For appropriate relative timescales of the dipolar couplings and ion transport processes, this is shown to be a facile method to compare ion dynamics among similar structures.

A quantitative assessment of oral contraceptive use and risk of ovarian cancer.

OBJECTIVE: To provide a quantitative assessment of the association between oral contraceptive (OC) use and ovarian cancer using results from the published literature. DATA SOURCES: We conducted a MEDLINE literature search for all epidemiologic studies of OC and ovarian cancer published in English between 1970-1991. The reference list for each article was reviewed to locate additional published articles. METHODS OF STUDY SELECTION: We included 20 studies in which a relative risk and either a standard error, confidence interval, or P value was reported, or sufficient data were presented to allow us to calculate these measures. DATA EXTRACTION AND SYNTHESIS: We summarized the findings using weighted averages and regression analyses. We found a summary relative risk of 0.64 (95% confidence interval 0.57-0.73) associated with ever-use of OC, indicating a 36% reduction in ovarian cancer risk. The risk of ovarian cancer decreased with increasing duration of OC use; we noted a 10-12% decrease in risk with 1 year of use and approximately a 50% decrease after 5 years of use. The reduced risk was present among both nulliparous and parous women and it appeared to last for at least 10 years after cessation of use. Although most studies assessed the use of cessation of use. Although most studies assessed the use of OC formulations from the 1960s and 1970s, data from the Cancer and Steroid Hormone Study indicate that the decreased ovarian cancer risk may also be present with current lower-dose formulations. CONCLUSION: The protective effect of OC against ovarian cancer risk should be considered in a woman's decision to use OC.

Electrophoretic profiles of Pasteurella multocida isolates from animals with hemorrhagic septicemia.

We determined that the protein profiles of 14 isolates from animals with hemorrhagic septicemia were relatively homogeneous and could be placed in 2 distinct groups on the basis of their country of origin. Such differences correlated with the serotypic properties of the individual isolates; hemorrhagic septicemia isolates of Asian and North American origin (Carter B) had a major protein band with an apparent molecular mass of 32 kDa, whereas those of African origins (Carter E) had a major protein band with an apparent molecular mass of 37 kDa. The possession of a major 32-kDa protein band appeared to be unique to Carter B isolates, suggesting that electrophoresis may be a useful nonserologic technique for the identification of organisms of this serotype. Other major bands with apparent molecular masses of 27, 45, and 47 kDa were shared by all strains, regardless of their serotype. The lipopolysaccharides were of low molecular mass and relatively uniform from 1 isolate to the next.

Haemorrhagic septicaemia: correlation of vaccinal antibody responses in mice with protection against Pasteurella multocida strain M1404.

This study examined the protection induced by oil adjuvant vaccine and broth bacterin in mice. Protective immunity was induced by both oil adjuvant and bacterin vaccination procedures. Oil adjuvant vaccination induced a 10(5)-fold increase for lethal challenge over control mice, while secondary vaccination induced a further 10-fold increase in resistance to lethal challenge. Broth bacterin induced a slightly weaker protective response with 10(4)- and 10(5)-fold increases in resistance to lethal challenge following primary and secondary vaccination, respectively. There was a significant relationship between IgG antibody levels and resistance to challenge (P = 0.026). Protection lasted for at least 20 weeks after a primary oil adjuvant vaccination. There was also a strong and significant relationship between IgG antibody levels and the passive protection afforded by serum transfer in each experiment within this study and the overall correlation was highly significant (P = 0.00001). There appeared to be a relationship between protection and the antibody response to major protein bands with the apparent molecular mass Mr. 94,000; 80,000; 67,000; 35,000 and 32,000 as well as to the bands in the region of the lipopolysaccharide components of P. multocida (approximately Mr, 14-15,000). Whether protection resulted from recognition of specific antigens or was a result of both antibody levels and antibody specificity remains to be defined.

Evidence of phenotypic dichotomy within an individual Pasteurella multocida type strain and among some haemorrhagic septicaemia-causing field isolates.

Haemorrhagic septicaemia-causing strains of Pasteurella multocida were identified by a disease-specific ELISA. Some strains, however, were of the same serotype as those which cause haemorrhagic septicaemia (HS) but were negative when tested in the disease specific ELISA. The suspect false negative isolates were passaged in mice and retested in the HS ELISA with the same result. Immunoelectron microscopy was used to examine further these suspect HS-causing strains. Monoclonal antibodies and protein A-gold showed that the suspect negative organisms were a mixture of phenotypes with less than 10 per cent, and usually less than 2 per cent, of the population expressing HS-associated epitopes. The degree of staining on the organisms expressing the HS-epitopes was of the same intensity as the positive control organism. Expression of the HS-associated epitopes is presumably too low to allow detection in the current HS ELISA.

Rapid identification of Pasteurella multocida organisms responsible for haemorrhagic septicaemia using an enzyme-linked immunosorbent assay.

Haemorrhagic septicaemia (HS) is caused by specific serotypes of Pasteurella multocida and is one of the major economic diseases of cattle and buffalo in South East Asia. Definitive diagnosis of the disease-causing organism with the available methods is labour intensive and not totally reliable, consequently, an ELISA system to identify P multocida organisms which cause HS was developed. One hundred and twenty-four P multocida isolates were tested, 58 were type strains and 66 were field isolates. Analysis of these strains indicated the assay had a specificity of 99 per cent and sensitivity of at least 86 per cent. The sensitivity could be an underestimate, as five isolates assumed to be false negative reactions may not all be HS-causing strains. The HS ELISA provides a rapid, simple, accurate and inexpensive diagnostic assay for identification of HS causing organisms but does not represent a new typing system for P multocida. This assay will also enable countries to assess the impact of HS more accurately.

Pasteurella multocida infections in mice with reference to haemorrhagic septicaemia in cattle and buffalo.

Haemorrhagic septicaemia (HS) is an infectious disease of cattle and buffalo caused by particular serotypes of Pasteurella multocida and is one of the most economically important livestock diseases in South-East Asia. While HS has been recognized for many years, very little is understood about the disease, primarily because of the expense of cattle and a lack of suitable animal models. The suitability of using mice to study HS was assessed using parameters such as the critical pathogenic dose, kinetics of infection, pathology of disease and resistance to reinfection. Pasteurella multocida M1404, the type strain for Carter group B, the serotype responsible for Asian HS, was injected intraperitoneally into BALB/c mice. As few as 20 colony forming units produced an overwhelming septicaemia in mice in less than 30 h. The kinetics of infection demonstrated a very rapid in vivo multiplication rate. There was no evidence of inhibition of bacterial cell growth by natural host defence mechanisms, even with the very small inocula used. The gross pathology of the disease in mice was characterized by splenomegaly, lymphadenopathy and petechial haemorrhages similar to that observed in cattle and buffalo with HS. Mice were found to develop a short-lived resistance to reinfection following a primary infection which had been successfully treated with antibiotics. The mouse would seem to provide an ideal tool by which to study HS, but warrant further studies in order to be able to critically assess it as a model for this economically important disease.

Evaluation of bovine antibody responses to haemorrhagic septicaemia vaccine.

ELISA and immunoblotting techniques were used to examine the humoral immune response to Pasteurella multocida, in bovine sera from Indonesia and Malaysia. Elevated levels of antibody to a crude lipopolysaccharide preparation were found in vaccinated animals. In addition to the response to lipopolysaccharide, antibodies from the vaccinated cattle strongly labelled five to six of the 40 protein bands in this organism.

The isolation of nucleic acid from nematodes requires an understanding of the parasite and its cuticular structure.

As parasitology increasingly becomes the domain of molecular biologists, it is important to keep in mind that a fundamental understanding of the whole parasite, its structure and behaviour can help to solve complex problems of molecular biology. Hugh Dawkins and Terence Spencer discuss the preparation of DNA from filaform larvae of Trichostrongylus colubriformis and Ostertagia circumcincta and how a detailed knowledge of the morphology and life cycle of each parasite helps in this process.

Antibody responses of sheep vaccinated with foot rot vaccines.

Enzyme-linked immunosorbent assay (ELISA) and crossed immunoelectrophoresis (IEP) were used to investigate antibody responses of sheep vaccinated with a double adjuvanted or single adjuvanted commercial foot rot vaccine. ELISA detected an antibody response of greater magnitude to the double adjuvant vaccine compared with the single adjuvant vaccine. Sera from sheep vaccinated with double adjuvant vaccine recognised at least six antigens of Bacteroides nodosus in crossed IEP while sera from the single adjuvant vaccinated sheep recognised one antigen. The use of non-denatured antigens of B nodosus in ELISA and crossed IEP enabled quantitative comparisons of antibody responses to the different foot rot vaccines to be made.

A technique for the purification of Mycobacterium paratuberculosis from the ileal mucosa of infected cattle.

Mycobacterium paratuberculosis, the causative agent of paratuberculosis, produces considerable economic loss in the cattle industry in many countries. The slow growth of M. paratuberculosis has hindered investigations of the antigenic composition of the organism and the development of species-specific antigen for serological detection of this disease. This paper describes a simple method for the isolation of large quantities of viable M. paratuberculosis from the intestinal mucosa of infected cattle by a combination of trypsin digestion, deoxyribonuclease/lysozyme treatment and differential centrifugation. Purity was about 99% and yield between 10(5)-10(9) bacteria/g tissue.

A reappraisal of the complement fixation test using soluble Mycobacterium avium antigen for the detection of M. paratuberculosis infection in cattle.

Serums from 263 cattle suspected of having paratuberculosis on the basis of clinical signs, were tested for antibodies to Mycobacterium paratuberculosis with a complement fixation test (CFT) employing a heat extracted, soluble M. avium antigen. Microscopic examination confirmed that 172 (65.4%) clinically affected animals had paratuberculosis, the remainder being disease-free. The specificity and sensitivity of the CFT was 92.3% and 74.4% respectively. Phenol treatment of serums before testing was compared with no treatment and was found to have no significant effect on the CFT titres. Results obtained are discussed in relation to the cause of false negative and false positive reactions.

Experimental infection of goats with Brucella ovis.

Six goats were inoculated with Brucella ovis. Two goats were inoculated with infected semen by the intratesticular route and 2 each by installation of the semen on to the nasal or preputial epithelium. All goats produced antibody responses as measured by an enzyme-linked immunosorbent assay (ELISA) procedure and the serums of 5 goats reacted in complement fixation tests for B. ovis. The 2 goats inoculated by the intratesticular route and one receiving B. ovis instilled intranasally subsequently excreted B. ovis in their semen. The possibility of natural transmission is discussed.

Differentiation of raw meat from phylogenically related species by enzyme-linked immunosorbent assay.

A method, based on enzyme-linked immunosorbent assay (ELISA), has been developed for differentiating raw meat from closely related species of economic importance. By visual assessment 0·1% donkey in horse, 0·1% goat in sheep and 1% buffalo in beef may be detected. The technique is rapid and simple to perform and could be used in abattoirs and coldstores with results being available within 1 h.

Enzyme-linked immunosorbent assay for Brucella ovis specific antibody in ram sera.

An enzyme-linked immunosorbent assay (ELISA) for the detection of Brucella ovis specific antibody in ram serum was compared with the currently employed complement fixation test (CFT). Rabbit anti-sheep IgG coupled to horseradish peroxidase was used as the antibody-enzyme conjugate and 2,2'-azino-di[3-ethylbenzthiazolin sulphonate (6)] as the substrate. The ratio of the optical density at 414 nm for positive and negative control sera (P/N ratio) was used to optimise the parameters of the test. Ram serum samples (16,527) were tested using ELISA and CFT (warm and cold) over a one year period. The ELISA was more sensitive and provided a more reliable measure of B ovis specific antibody than did the CFT. Implications of employing ELISA as the sole test in an eradication scheme are discussed.

Stable enhancement of calcium retention in mitochondria isolated from rat liver after the administration of glucagon to the intact animal.

1. Mitochondria isolated from rat liver by centrifugation of the homogenate in buffered iso-osmotic sucrose at between 4000 and 8000g-min, 1h after the administration in vivo of 30mug of glucagon/100g body wt., retain Ca(2+) for over 45min after its addition at 100nmol/mg of mitochondrial protein in the presence of 2mm-P(i). In similar experiments, but after the administration of saline (0.9% NaCl) in place of glucagon, Ca(2+) is retained for 6-8min. The ability of glucagon to enhance Ca(2+) retention is completely prevented by co-administration of 4.2mg of puromycin/100g body wt. 2. The resting rate of respiration after Ca(2+) accumulation by mitochondria from glucagon-treated rats remains low by contrast with that from saline-treated rats. Respiration in the latter mitochondria increased markedly after the Ca(2+) accumulation, reflecting the uncoupling action of the ion. 3. Concomitant with the enhanced retention of Ca(2+) and low rates of resting respiration by mitochondria from glucagon-treated rats was an increased ability to retain endogenous adenine nucleotides. 4. An investigation of properties of mitochondria known to influence Ca(2+) transport revealed a significantly higher concentration of adenine nucleotides but not of P(i) in those from glucagon-treated rats. The membrane potential remained unchanged, but the transmembrane pH gradient increased by approx. 10mV, indicating increased alkalinity of the matrix space. 5. Depletion of endogenous adenine nucleotides by P(i) treatment in mitochondria from both glucagon-treated and saline-treated rats led to a marked diminution in ability to retain Ca(2+). The activity of the adenine nucleotide translocase was unaffected by glucagon treatment of rats in vivo. 6. Although the data are consistent with the argument that the Ca(2+)-translocation cycle in rat liver mitochondria is a target for glucagon action in vivo, they do not permit conclusions to be drawn about the molecular mechanisms involved in the glucagon-induced alteration to this cycle.

The transport and oxidation of succinate by Ehrlich ascites-tumour cells.

The transport and oxidation of succinate by functionally intact Ehrlich ascites-tumour cells was investigated. On the basis of pH dependence and inhibitor sensitivity it was concluded that succinate may be transported across the cell membrane by the organic anion carrier system. Thus the ability of isolated Ehrlich cells to oxidize succinate is real, and is not necessarily a result of damage to cell integrity.