Application of the Three Rs to challenge assays used in vaccine testing: Tenth report of the BVAAWF/FRAME/RSPCA/UFAW Joint Working Group on Refinement

This report aims to facilitate the implementation of the Three Rs (reduction, refinement and replacement) in the testing of vaccines for regulatory and other purposes. The focus is predominantly on identification of reduction and refinement opportunities in batch potency testing but the principles described are widely applicable to other situations that involve experimental infections of animals. The report should also help to interpret the requirements of the European Pharmacopoeia with regard to the use of alternative tests, humane endpoints and other refinements. Two specific worked examples, for batch potency testing of Clostridium chauvoei and canine leptospira, with recommendations for harmonisation of international test requirements for these and other vaccines, are provided as appendices online.     

Motility of isolated mammalian gastric fundus

Gastric fundus strips (GFS) of rats, guinea-pigs, rabbits, cats and humans, but not of dogs, show spontaneous phasic contractions in vitro. Rabbit, cat, dog and human GFS exhibit dose-dependent responses to acetylcholine (ACh), histamine (His), serotonin (5-HT) and KCl. Guinea-pig drug-induced responses are not dose-dependent. Rat GFS does not respond to His. ED50S for KCl are always higher than for other agonists. ED50S for 5-HT are lower than for other drugs in rats and cats, but not different in other species. Maximal GFS contractions induced by ACh are usually higher than those induced by other drugs.     

c-myc gene sequences and the phylogeny of bats and other eutherian mammals

The complete protein-coding sequences of the c-myc proto-oncogene were determined for five species of four new orders of eutherian (placental) mammals. These newly obtained sequences were aligned to each other and to other available orthologs for the phylogenetic estimation of eutherian interordinal relationships. Several measures of sequence difference and base composition were first calculated to assess the major evolutionary properties of the three codon positions and two protein-coding exons of the gene. On the basis of these calculations, different parsimony, distance, and maximum likelihood approaches were adopted, with the most sophisticated involving the separate, then combined, likelihood analyses of the third codon positions of exon 2 versus all other sites. These phylogenetic approaches provided clear support for the grouping of Chiroptera (bats) with Artiodactyla (ruminants, camels, and pigs) and Carnivora (cats, dogs, and their allies), an interordinal arrangement that receives strong corroboration from other lines of evidence including complete mitochondrial DNA sequences. In contrast, these analyses failed to provide strong to reasonable support for any other interordinal group. This study concludes with specific recommendations about sampling and other strategies for maximizing the phylogenetic contributions of the c-myc gene to the continued resolution of the eutherian ordinal tree.     

Assessing rbf networks using DELVE

In this paper, different methods for training radial basis function (RBF) networks for regression problems are described and illustrated. Then, using data from the DELVE archive, they are empirically compared with each other and with some other well known methods for machine learning. Each of the RBF methods performs well on at least one DELVE task, but none are as consistent as the best of the other non-RBF methods.     

Identification of Rhodococcus equi using the polymerase chain reaction

K.S. BELL, J.C. PHILP, N. CHRISTOFI AND D.W.J. AW. 1996. Two regions in the gene coding for 16S rRNA in Rhodococcus equi were selected as species-specific primer sequences for the polymerase chain reaction (PCR). PCR using these primers was tested against 10 strains of R. equi (including the type strain) and gave positive results for all but was negative for all other tested species of Rhodococcus ; representatives of the most closely related genera and a number of other bacterial species. This method could therefore be used to identify this species which can infect the lungs or other organs of horses, pigs, humans and other animals.     

Serial-Section Analysis of Clustering within Anthers of Maize Microsporocytes with Specific Crossovers

The distribution within anthers of maize plants of microsporocytes of differing crossover class was studied by application of the serial-sectioning technique to plants heterozygous for a paracentric inversion. In such material, cells with four-strand double crossovers within the inversion are distinguishable from cells with other classes of crossovers within the inverted region and from cells with no crossovers within the inversion. Evidence was found for coarse clustering of cells with four-strand double crossovers within the inversion and for coarse clustering of cells of other crossover classes (with at least one crossover within the inversion). The local frequencies of these two major categories of crossover cells appeared to vary independently of each other, with regional increases in each occurring at the expense of noncrossover cells. There is also some suggestion that cells with four-strand doubles within the inversion may be clustered independently of other double-crossover classes within the inversion, but these other classes are not directly scorable.     

Legionella oakridgensis: unusual new species isolated from cooling tower water.

We describe a new species of Legionella represented by 10 strains isolated from industrial cooling towers. Legionella oakridgensis differed genetically from the other seven species of Legionella in DNA hybridization studies and differed serologically in direct fluorescent-antibody tests. The new species, unlike all other species except L. jordanis, did not require added L-cysteine for growth in serial transfer on charcoal-yeast extract agar. L. oakridgensis, as well as three other species tested, required L-cysteine for primary isolation from animal tissues. L. oakridgensis was the only species of Legionella that failed to produce alkaline phosphatase at pH 8.5. In all other respects, it resembled other species of Legionella, including having a high content of branched-chain cellular fatty acids and being pathogenic for guinea pigs. These bacteria have not yet been associated with human disease, but they are potential causes of legionellosis.     

Structure-activity studies on halluncinogenic amphetamines using model interaction calculations

In summary, we can briefly describe the model as developed at this time. We view the amphetamine receptor as possessing two principal features: one site provides anchoring for the onium group, whereas the other feature interacts with the phenyl ring and substitutents. In addition, there is implied the presence of a feature associated with the 3,4 region of the phenyl ring. The best model for the phenyl ring receptor is the methylindole molecule with the two rings, one directly above the other at a separation distance of 4·5 Å. The correlation from these results [equation (2)] is better than that reported for any other SAR study to date, accounting for 74 % of the variation in the data. Further inclusion of a term relating to distribution or absorption improves the correlation so that 82Y. of the variation is accounted for.This provisional model can now be tested against data for other amphetamines, mescalines, tryptamines and other hallucinogenic agents.     

Partnership

Individuals are called partners when it is in their best interest to help each other, if by doing so they increase the probability of being together in the future when, for similar reasons, they will continue to help each other. Kinsmen or individuals who often face (hedonic) situations in which helping is the dominating strategy are committed to help each other. Partnership may develop among them since the loss of the other means the loss of a guaranteed helper. Thus, they may be willing to take additional risks to help each other. Partnership may occur among unrelated individuals and with no hedonic situations. Partnership creates bonds between partners which may be much stronger than those between kinsmen; an individual may take more risks for his partner than he will ever take for a kin. Partnership may evolve without the sophistication and memory required for reciprocation altruism. Although kin selection, partnership and reciprocation are likely to appear fused as the causes for altruism, we argue that it may be possible to distinguish between them in some situations. We show that as the partners get older partnership may become less important to them. We also show that like cooperation, and for analogous reasons, malice may evolve among partners so that each will be willing to take additional risks in order to eliminate the other.     

Prey-capture techniques and prey preferences of Habrocestum pulex, an ant-eating jumping spider (Araneae, Salticidae) from North America

The prey-catching techniques and prey preferences of Habrocestum pulex (Hentz), ant-eating jumping spider (Araneae: Salticidae) from North America, were studied in the laboratory. H. pulex uses prey-specific, prey-catching behaviour against ants. Ants, but not other insects, were consistently attacked head-on. After attacking an ant, but not after attacking other insects, H. pulex kept its forelegs extended laterally and forwards without touching the ground. H. pulex feeds on ants in preference to other insects. Preference for ants and prey-specific predatory behaviour do not depend on prior experience with ants. As in earlier studies of other ant-eating salticids, three different types of tests for prey preference were carried out, using active, living prey: Type 1 (one type of prey presented to salticid at a time on alternate days); Type 2 (two types of prey presented to salticid simultaneously); and Type 3 (salticid feeding on one type of prey presented with alternative prey of another type). However, newly-designed apparatus made testing more efficient. Preference for ants over other insects is shown not to depend on level of activity or any other cues from prey movement pattern: Type 1 and Type 2 tests were carried out using motionless (dead) lures, and again ants were taken in preference to other insects. Findings from this study are discussed in relation to recent findings on other ant-eating salticids.     

Contraceptive practices and fertility among Southeast Asian, black, and white mothers attending a maternal infant care program

The purpose of this analysis was to determine if there were differences in selected fertility characteristics including parity, pregnancy spacing, age at 1st pregnancy, age of menarche, breastfeeding postpartum, and contraceptive practices among white, black, Hmong, and other Southeast Asian mothers attending a maternal infant care program in Minneapolis, Minnesota, during 1980-1982. White and black mothers were younger than the Hmong and other Asian mothers. The lowest mean age of 1st pregnancy was among blacks. Ages of 1st pregnancy were similar for whites, Hmong, and other Asians, although the mean age of menarche was approximately 2 years later for Hmong and other Southeast Asian mothers compared to the white and black mothers. Based on self reports at the 1st postpartum visit 1 month after delivery, 39% of the whites and 25% of the blacks were breastfeeding. In comparison 8.7% of the Hmong and 17% of the other southeast Asian mothers were breastfeeding. Maternal aged age at 1st pregnancy were significant predictors of parity for whites, blacks, Hmong, and other Southeast Asians. Ever-use of contraception was significant predictor of parity only for Hmong. The highest proportion of ever-users of contraception was among the white mothers (80%) followed by the blacks (69.3%) and other Asian mothers (34.85). Hmong mothers had the lowest proportion having used contraception (17.1%).     

Use of wood energy in the United States — An opportunity

The use of wood for energy — including the burning of solid wood and black liquor from pulping — has been growing at a rate significantly greater than that for all other uses such as lumber, pulp, or particleboard. In the United States, the end of most wood is not lumber or pulp and paper but feed for energy. In 1983, 155·5 M Mg of wood were used for energy. This could threaten to increase the price of wood for those other uses, or it can stimulate us to seek more creative ways of using untapped wood resources for fuel.On the basis of estimates of heavy wood energy use relative to other uses for wood, and estimates of continuing high costs for fossil fuels, we suggest here the feasibility of meeting the demand for fuelwood through small-scale cooperatives. Such an approach can improve forestry practices and can avoid unduly increasing the cost of wood for other end uses.     

Patterns of clometacillin circulation in the body of laboratory animals]

Distribution of clometacillin in mice, rats and rabbits was studied in comparison with some other penicillins. It was found that clometacillin was superior to all other penicillins used for comparison with respect to the circulation time in the blood after intravenous, intramusclar or intrastomach administration. As for the capacity for penetrating into the tissues from the blood, clometacillin was not inferior to ampicillin and benzylpenicillin and was superior to propicillin, though it was bound by the blood serum proteins to a greater extent than the other penicillins used for comparison.     

Analysis of the seasonality of acute intestinal diseases with regard to the detection of dysentery by antigen indication

Coincidence in the seasonal changes of the registered morbidity in dysentery and in other acute intestinal diseases is observed. The occurrence of Shigella antigens, detected with the use of erythrocyte diagnostic reagents, in the excreta of patients with the clinical diagnosis of dysentery and patients with other acute intestinal disease has a seasonal character, its peak coinciding with the period of increased morbidity in bacteriologically confirmed dysentery. The correction of monthly morbidity levels in dysentery and in other acute intestinal diseases with due regard for the proportion of Shigella antigens in the findings, made in the groups of patients with the clinical diagnosis of dysentery and with the diagnosis of other acute intestinal diseases for the corresponding month, has revealed that the actual morbidity in other acute intestinal diseases has a less pronounced seasonal character in comparison with the registered morbidity. This fact substantiates the statement that in the group of patients with other acute intestinal diseases a large proportion of such diseases is, actually, of a noninfectious nature.     

The rabbit differs from other mammalian in the tissue distribution of alkaline phosphatase isoenzymes

There are only two gene loci code for alkaline phosphatase of mammalian other than human and great apes: one for the intestinal form and other for the liver/kidney/bone form. The former form is present only in the intestine and the latter form occurs in other tissues such as liver, kidney and bone. In the present study, the rabbit was found to be different from other mammalian in the tissue distribution of alkaline phosphatase isoenzymes: only in the rabbit, most of the enzyme in the kidney and liver was the third form which differs from the liver/kidney/bone form, and this form was enzymatically and immunologically similar to the intestinal form of ALPase.     

FIELD EXPERIMENTS SHOW THAT ACOUSTIC PINGERS REDUCE MARINE MAMMAL BYCATCH IN THE CALIFORNIA DRIFT GILL NET FISHERY

A controlled experiment was carried out in 1996–1997 to determine whether acoustic deterrent devices (pingers) reduce marine mammal bycatch in the California drift gill net fishery for swordfish and sharks. Using Fisher's exact test, bycatch rates with pingers were significantly less for all cetacean species combined ( P < 0.001) and for all pinniped species combined ( P = 0.003). For species tested separately with this test, bycatch reduction was statistically significant for short-beaked common dolphins ( P = 0.001) and California sea lions ( P = 0.02). Bycatch reduction is not statistically significant for the other species tested separately, but sample sizes and statistical power were low, and bycatch rates were lower in pingered nets for six of the eight other cetacean and pinniped species. A log-linear model relating the mean rate of entanglement to the number of pingers deployed was fit to the data for three groups: short-beaked common dolphins, other cetaceans, and pinnipeds. For a net with 40 pingers, the models predict approximately a 12-fold decrease in entanglement for short-beaked common dolphins, a 4-fold decrease for other cetaceans, and a 3-fold decrease for pinnipeds. No other variables were found that could explain this effect. The pinger experiment ended when regulations were enacted to make pingers mandatory in this fishery.     

2009年全国其他感染性腹泻报告病例分析

目的了解我国2009年其他感染性腹泻的流行特点和病原学信息,提出改善建议,为更好的控制其他感染性腹泻病的流行提供数据支持。方法收集中国疾病控制信息系统"疾病监测信息报告管理系统"中2009年其他感染性腹泻的病例报告,对其流行病学及病原学信息进行描述性统计分析。结果 2009年全国共报告655 965例其他感染性腹泻病例,其中≤5岁儿童分别占报告总病例的50.38%和死亡病例的70.0%。实验室确诊病例占报告病例数的5.04%,其中病毒性感染占92.79%,细菌性感染占6.92%。结论≤5岁的散居儿童应为其他感染性腹泻的重点监测人群。其他感染性腹泻报告病例中约有95%为临床诊断病例。我国其他感染性腹泻的病原学诊断率及病原学诊断结果的报告率亟待提高。     

Caffeine and other xanthines as cytochemical blockers and removers of heterocyclic DNA intercalators from chromatin

Caffeine (CAF) and other xanthines non-covalently bind with the cationic fluorescent dye acridine orange (AO) and with other heterocyclic mutagens and carcinogens that are known to intercalate into double-stranded DNA (dsDNA). Fluorescence microscopy and spectrofluorometry studies were employed to test the ability of caffeine and certain other methyl substituted xanthines, with different binding affinities for AO, to inhibit and to reverse the intercalation of AO and other heterocyclic agents from intercalation with the DNA of nuclear chromatin of air-dried cells. Results indicated that xanthines with binding affinity for AO greater than 150 m(-1) block the AO molecule in a concentration dependent manner and comply with mass action kinetics. Thus CAF and other xanthines can be used to either inhibit intercalation of AO into nuclear DNA or to remove AO once intercalated into nuclear DNA. The interactions between other planar heterocyclics, xanthines, and nuclear chromatin dsDNA were also found to be non-covalent. Studies are needed to determine the ability of CAF and other xanthines to block and/or remove polyaromatic hydrocarbon (PAH) intercalators from the DNA of living cells.     

Enhanced transport of natural amino acids after activation of pig lymphocytes.

Na+-dependent uptake of the amino acids L-proline and L-methionine was greatly accelerated when pig lymphocytes were activated with phytohaemagglutinin or other mitogens. The increased influx was apparent after incubation with phytohaemagglutinin for 1 h, and reached a maximum after 24 h. The lymphocytes appear to possess at least three different transport systems for neutral amino acids with properties similar to, but not identical with, those described for other cells. The activity of a system resembling the A system of other cells was increased most dramatically after activation, its activity in unstimulated lymphocytes being extremely low or absent. A second Na+-dependent system, which has properties similar to those of the ASC system in other cells, but with a broader specificity for amino acids, was more active in unstimulated lymphocytes, and uptake by this system was also accelerated after incubation with phytohaemagglutinin. The activity of a third system, very similar to the L system in other cells, was increased to a much smaller extent after lymphocyte activation.