A combination of cis and trans control can solve the hotspot conversion paradox.

There is growing evidence that in a variety of organisms the majority of meiotic recombination events occur at a relatively small fraction of loci, known as recombination hotspots. If hotspot activity results from the DNA sequence at or near the hotspot itself (in cis), these hotspots are expected to be rapidly lost due to biased gene conversion, unless there is strong selection in favor of the hotspot itself. This phenomenon makes it very difficult to maintain existing hotspots and even more difficult for new hotspots to evolve; it has therefore come to be known as the "hotspot conversion paradox." I develop an analytical framework for exploring the evolution of recombination hotspots under the forces of selection, mutation, and conversion. I derive the general conditions under which cis- and trans-controlled hotspots can be maintained, as well as those under which new hotspots controlled by both a cis and a trans locus can invade a population. I show that the conditions for maintenance of and invasion by trans- or cis-plus-trans-controlled hotspots are broader than for those controlled entirely in cis. Finally, I show that a combination of cis and trans control may allow for long-lived polymorphisms in hotspot activity, the patterns of which may explain some recently observed features of recombination hotspots.

High frequency of cryptic deleterious mutations in Caenorhabditis elegans.

Deleterious mutations with very small phenotypic effects could be important for several evolutionary phenomena, but the extent of their contribution has been unknown. Fitness effects of induced mutations in lines of Caenorhabditis elegans were measured using a system for which the number of deleterious point mutations in the DNA can be estimated. In fitness assays, only about 4 percent of the deleterious mutations fixed in each line were detectable. The remaining 96 percent, though cryptic, are significant for mutation load and, potentially, for the evolution of sex.

A test for epistasis among induced mutations in Caenorhabditis elegans.

Synergistic epistasis, in which deleterious mutations tend to magnify each other's effects, is a necessary component of the mutational deterministic hypothesis for the maintenance of sexual production. We tested for epistasis for life-history traits in the soil nematode Caenorhabditis elegans by inducing mutations in two genetic backgrounds: a wild-type strain and a set of genetically loaded lines that contain large numbers of independent mildly detrimental mutations. There was no significant difference between the effect of new mutations on the wild-type background and the genetically loaded background for four out of five fitness correlates. In these four cases, the maximum level of epistasis compatible with the data was very low. The fifth trait, late productivity, is not likely to be an important component of fitness. This suggests either that specific environmental conditions are required to cause epistasis or that synergistic epistasis is not a general phenomenon. We also suggest a new mechanism by which deleterious mutations may provide an advantage to sexual reproduction under low selection coefficients.

Testing for epistasis between deleterious mutations.

Determining the way in which deleterious mutations interact in their effects on fitness is crucial to numerous areas in population genetics and evolutionary biology. For example, if each additional mutation leads to a greater decrease in log fitness than the last (synergistic epistasis), then the evolution of sex and recombination may be favored to facilitate the elimination of deleterious mutations. However, there is a severe shortage of relevant data. Three relatively simple experimental methods to test for epistasis between deleterious mutations in haploid species have recently been proposed. These methods involve crossing individuals and examining the mean and/or skew in log fitness of the offspring and parents. The main aim of this paper is to formalize these methods, and determine the most effective way in which tests for epistasis could be carried out. We show that only one of these methods is likely to give useful results: crossing individuals that have very different numbers of deleterious mutations, and comparing the mean log fitness of the parents with that of their offspring. We also reconsider experimental data collected on Chlamydomonas moewussi using two of the three methods. Finally, we suggest how the test could be applied to diploid species.

Dominance and overdominance of mildly deleterious induced mutations for fitness traits in Caenorhabditis elegans.

We estimated the average dominance coefficient of mildly deleterious mutations (h, the proportion by which mutations in the heterozygous state reduce fitness components relative to those in the homozygous state) in the nematode Caenorhabditis elegans. From 56 worm lines that carry mutations induced by the point mutagen ethyl methanesulfonate (EMS), we selected 19 lines that are relatively high in fitness and estimated the viabilities, productivities, and relative fitnesses of heterozygotes and homozygotes compared to the ancestral wild type. There was very little effect of homozygous or heterozygous mutations on egg-to-adult viability. For productivity and relative fitness, we found that the average dominance coefficient, h, was approximately 0.1, suggesting that mildly deleterious mutations are on average partially recessive. These estimates were not significantly different from zero (complete recessivity) but were significantly different from 0.5 (additivity). In addition, there was a significant amount of variation in h among lines, and analysis of average dominance coefficients of individual lines suggested that several lines showed overdominance for fitness. Further investigation of two of these lines partially confirmed this finding.

Properties of ethylmethane sulfonate-induced mutations affecting life-history traits in Caenorhabditis elegans and inferences about bivariate distributions of mutation effects.

The homozygous effects of ethylmethane sulfonate (EMS)-induced mutations in Caenorhabditis elegans are compared across life-history traits. Mutagenesis has a greater effect on early than late reproductive output, since EMS-induced mutations tend to cause delayed reproduction. Mutagenesis changes the mean and variance of longevity much less than reproductive output traits. Mutations that increase total or early productivity are not detected, but the net effect of mutations is to increase and decrease late productivity to approximately equal extents. Although most mutations decrease longevity, a mutant line with increased longevity was found. A flattening of mortality curves with age is noted, particularly in EMS lines. We infer that less than one-tenth of mutations that have fitness effects in natural conditions are detected in the laboratory, and such mutations have moderately large effects ( approximately 20% of the mean). Mutational correlations for life-history traits are strong and positive. Correlations between early or late productivity and longevity are of similar magnitude. We develop a maximum-likelihood procedure to infer bivariate distributions of mutation effects. We show that strong mutation-induced genetic correlations do not necessarily imply strong directional correlations between mutational effects, since correlation is also generated by lines carrying different numbers of mutations.

Edinburgh drug users: are they injecting and sharing less?

OBJECTIVE: To describe epidemiological trends in injecting and equipment sharing among a sample drawn from a drug-using population with a high rate of HIV infection. DESIGN: A structured interview was administered prior to treatment to cross-sectional samples of drug users over the period 1988-1991. Reports of injecting, sharing and HIV infection were compared annually. SETTING: Lothian Health Board's Community Drug Problems Service is a secondary level service offering harm reduction and treatment of dependency. PARTICIPANTS: A total of 734 consecutively referred drug users resident in Edinburgh. Re-referrals in the same calendar year were excluded. MEASURES: History of injecting and sharing, recent injecting and sharing, HIV testing history, drugs used in previous month and substitute prescription status. RESULTS: Large reductions in the frequency of injecting were found over the 4 years even among those who were not receiving oral substitutes. More participants in latter years were receiving prescriptions combining opioids and benzodiazepines. Fewer of those interviewed latterly had ever shared injecting equipment. Among recent injectors just as many share equipment as previously. HIV prevalence did not vary significantly over the period. An HIV prevalence of 19% was reported among recent injectors. CONCLUSIONS: Edinburgh's drug users are engaging in far safer drug-taking behaviour than previously. Levels of HIV in this population suggest that the epidemic is being contained. A small number persist in high risk drug-related activities. Further investigation of the characteristics of these individuals and the need to develop novel methods of influencing their behaviour are recommended.

The Red Queen and Fluctuating Epistasis: A Population Genetic Analysis of Antagonistic Coevolution.

Host-parasite coevolution has been shown to provide an advantage to recombination, but the selective mechanism underlying this advantage is unclear. One possibility is that recombination increases the frequency of advantageous genotypes that are disproportionately rare because of fluctuating epistasis. However, for this mechanism to work, epistasis for fitness must fluctuate over a very narrow timescale: two to five generations. Alternatively, recombination may speed up the response to directional selection by breaking up linkage disequilibria that decrease additive genetic variance. Here we analyze the results of a numerical simulation of host-parasite coevolution to assess the importance of these two mechanisms. We find that linkage disequilibria may tend to increase, rather than decrease, additive genetic variance. In addition, the sign of epistasis changes every two to five generations under several of the parameter values investigated, and epistasis and linkage disequilibrium are frequently of opposite signs. These results are consistent with the idea that selection for recombination is mediated by fluctuating epistasis. Finally, we explore the conditions under which an allele causing free recombination can spread in a nonrecombining host population and find general agreement between the predictions of a population genetic model of fluctuating epistasis and our simulation model.

Methadone treatment in the Scottish context: outcomes of a community-based service for drug users in Lothian.

Few studies investigating the effectiveness of methadone treatment for opiate dependence have emanated from the UK. The core feature of treatment offered by Lothian Health's Community Drug Problems Service involves the prescribing of methadone by the client's general practitioner. Of a cohort of 494 daily users of opiates attending the service, 39% remained in treatment for at least 12 months. Up to 2 years in-treatment follow-up revealed significant improvement in injecting and criminal behaviour. There were no HIV seroconversions reported during the treatment period. There was no improvement in injection equipment sharing, condom use, illicit drug use or employment status. 'Satisfactory' discharge was achieved for 40% of those in treatment for at least 6 months. These results are largely consistent with the outcomes of methadone programmes elsewhere.

Effect of starvation on parasite-induced mortality in a freshwater snail (Potamopyrgus antipodarum).

The level of host exploitation is expected, under theory, to be selected to maximise (subject to constraints) the lifetime reproductive success of the parasite. Here we studied the effect of two castrating trematode species on their intermediate snail host, Potamopyrgus antipodarum. One of the trematode species, Microphallus sp., encysts in the snail host and the encysted larvae "hatch" following ingestion of infected snails by birds. The other species, Notocotylus gippyensis, by contrast, releases swimming larvae; ingestion of the snail host is not required for, and does not aid, transmission to the final host. We isolated field-collected snails for 3 months in the laboratory, and followed the survival of infected and uninfected snails under two conditions: not fed and fed ad libitum. Mortality of the infected hosts was higher than mortality of the uninfected ones, but the response to starvation treatment was parasite species specific. N. gippyensis induced significantly higher mortality in starved snails than did Microphallus. Based on these results, we suggest that host exploitation by different species of trematodes may depend on the type of transmission. Encysting in the snail host may select for a reduced rate of host exploitation so as to increase the probability of transmission to the final host.

Effect of polymerization mode of a dual-cured resin cement on time-dependent shear bond strength to porcelain.

PURPOSE: To evaluate the shear bond strengths to porcelain and setting times of a dual-cured resin cement with light/chemical curing (dual) or chemical only curing versus time. MATERIALS AND METHODS: Variolink resin cement was bonded to specimens of etched, silanated porcelain. Groups of specimens were cured by dual cured or by chemical curing only. Shear bond strengths were recorded at 2, 3, 5, 60 minutes and 24 hours for dual cured and at 10, 20, 40, 50, 60 minutes and 24 hours for chemical-cured only after mixing. Kinetic-temperature profiles of dual and chemical curing modes of cement were calculated. RESULTS: Maximum bond strengths and time to attainment were 17.5 +/- 2.7 MPa at 60 minutes for chemical-cured and 26.1 +/- 2.3 MPa at 5 minutes for dual-cured. The peak in the kinetic-temperature setting profiles were 14.1 +/- 0.9 minutes for chemical curing and 52.2 +/- 5.2 seconds for dual curing. Dual curing provided significantly higher shear bond strengths versus chemical curing at both the 60-minute and 24-hour time periods.

Short- and long-term benefits and detriments to recombination under antagonistic coevolution.

We explored the evolution of recombination under antagonistic coevolution, concentrating on the equilibrium frequencies of modifier alleles causing recombination in initially nonrecombining populations. We found that the equilibrium level of recombination in the host depended not only on parasite virulence, but also on the strength of the modifier allele, and on whether or not the modifier was physically linked to the parasite interaction loci. Nonetheless, the maximum level of recombination for linked loci at equilibrium was about 0.3 (60% of free recombination) for interactions with highly virulent parasites; the level decreased for unlinked modifiers, and for lower levels of parasite virulence. We conclude that recombination spreads because it provides a combination of an immediate (next-generation) fitness benefit and a delayed (two or more generations) increase in the rate of response to directional selection. The relative impact of these two mechanisms depends on the virulence of parasites early in the spread of the modifier, but a trade-off between the two dictates the equilibrium modifier frequency for all nonzero virulences that we examined. In addition, population mean fitness was higher in populations at intermediate equilibria than populations fixed for free recombination or no recombination. The difference, however, was not enough on its own to overcome the two-fold cost of producing males.

A pilot survey of client opinion of the Lothian Community Drug Problem Service: likes, dislikes, efficacy and improvements.

An open ended unstructured questionnaire was used to find out the opinion of clients of the Community Drug Problem Service of Lothian Health Board, concerning how the service had helped them and for suggestions for improvement. A total of 63 questionnaires were returned over a six month period. Analysis showed that 77% of respondents valued counselling services but 26% complained of too low oral substitute drug dosage. Improvement in personal development, stabilised drug use, less criminality and stopping injecting were reported as being the main benefits. A more detailed self reporting questionnaire will be developed from this pilot study to answer some of the questions raised and for use as a routine part of ongoing service monitoring and evaluation.

Gender differences in HIV risk behaviour of injecting drug users in Edinburgh.

A multi-site sample of currently-injecting drug users (IDUs) comprising 344 men and 136 women was recruited in Edinburgh. Sixty-seven per cent of the sample said they had at some time used injecting equipment already used by another person and 25% admitted doing so in the 6 months before interview. Whereas women who injected with used equipment obtained it predominantly from a sexual partner, for men the source was more often a close friend or someone whose HIV status they were unlikely to know. In the 6 months before interview, 40% of men, compared with 20% of women, had more than one heterosexual partner. This difference was associated with a higher proportion of men with steady partners also having casual partners. Women IDUs were more likely to have regular partners who injected (57% vs 26%). Though sharing of injecting equipment has already diminished in Edinburgh, further measures are needed to eliminate it. For injectors here, the risk of infection from unprotected heterosexual intercourse may now be greater than that from sharing injecting equipment, particularly for women. Other methods of encouraging changes in sexual behaviour need to be investigated and successful ones promoted.

ENVIRONMENTAL STRESS AND THE MAINTENANCE OF SEX IN A FRESHWATER SNAIL.

Synergism among mutations can lead to an advantage to sexual reproduction, provided mutation rates are high enough (the mutational deterministic hypothesis). Here we tested the idea that competition for food can increase the advantage to sexual reproduction, perhaps by increasing the synergism among mutations in asexual individuals. We compared the survivorship of sexual and asexual snails (Potamopyrgus antipodarum) under two treatments: starved and fed. We predicted higher mortality for asexual snails when starved, but found that sexual and asexual individuals survived at the same rate, independent of treatment. These results suggest that the distribution of sex in this snail may not be explained by variation in competition among populations.