Summary1. How thermal tolerance estimated in the laboratory can be extrapolated to natural settings remains a contentious subject. Here, we argue that the general premise that a single temperature can accurately describe upper or lower tolerance limits is incorrect. 2. Survival probability is determined by both the intensity and the duration of a thermal stress, and the association between these variables can be adequately conveyed by a thermal tolerance landscape. Employing this framework, we demonstrate that the temperature range that an organism can tolerate is expected to narrow down with the duration of the thermal challenge. 3. Analyses suggest that a trade-off exists between tolerances to acute and chronic exposition to thermal stress, and that changes in temperature means or extremes may result in drastically different selective pressures and subsequent evolutionary responses. 4. After controlling for the duration of the thermal challenge, we also uncover latitudinal effects on upper lethal temperatures in insects that remained unnoticed in previous broad-scale comparative analyses. 5. Ultimately, critical thermal limits have been adopted in the ecological literature for logistic reasons and are inadequate descriptors of thermal tolerance on conceptual grounds. We consider that tolerance landscapes provide a more suitable framework to study temperature tolerance and its potential impact in ecological settings.

BackgroundOur current understanding of evolution is so tightly linked to template-dependent replication of DNA and RNA molecules that the old idea from Oparin of a self-reproducing 'garbage bag' ('coacervate') of chemicals that predated fully-fledged cell-like entities seems to be farfetched to most scientists today. However, this is exactly the kind of scheme we propose for how Darwinian evolution could have occurred prior to template replication.ResultsWe cannot confirm previous claims that autocatalytic sets of organic polymer molecules could undergo evolution in any interesting sense by themselves. While we and others have previously imagined inhibition would result in selectability, we found that it produced multiple attractors in an autocatalytic set that cannot be selected for. Instead, we discovered that if general conditions are satisfied, the accumulation of adaptations in chemical reaction networks can occur. These conditions are the existence of rare reactions producing viable cores (analogous to a genotype), that sustains a molecular periphery (analogous to a phenotype).ConclusionsWe conclude that only when a chemical reaction network consists of many such viable cores, can it be evolvable. When many cores are enclosed in a compartment there is competition between cores within the same compartment, and when there are many compartments, there is between-compartment competition due to the phenotypic effects of cores and their periphery at the compartment level. Acquisition of cores by rare chemical events, and loss of cores at division, allows macromutation, limited heredity and selectability, thus explaining how a poor man's natural selection could have operated prior to genetic templates. This is the only demonstration to date of a mechanism by which pre-template accumulation of adaptation could occur.ReviewersThis article was reviewed by William Martin and Eugene Koonin.

Summary1. Current studies indicate that estimates of thermal tolerance limits in ectotherms depend on the experimental protocol used, with slower and presumably more ecologically relevant rates of warming negatively affecting the upper thermal limits (CT max ). Recent empirical evidence also gives credence to earlier speculations suggesting that estimates of heritability could drop with slower heating rates. 2. Using published data from the fruit fly Drosophila melanogaster, we show that empirical patterns can be explained if flies' physical condition decreases with experimental time in thermal tolerance assays. This problem could even overshadow potential benefits of thermal acclimation, also suggesting that a drop in CT max with slower heating rates does not necessarily rule out beneficial acclimatory responses. 3. Numerical results from a simple illustrative model show that no clear conclusions can be obtained on how the phenotypic variance in CT max will be affected with different rates of thermal change. Conversely, the genetic variance and estimated heritabilities are expected to drop with slower heating rates, hence ramping rates in experiments aiming to study the evolutionary potential of thermal tolerance to respond to global warming should be as fast as possible (within the range in which measurement accuracy and physical condition are not affected). 4. Measurements under ecologically realistic warming rates should also consider the impact of other physiological and behavioural strategies that might partly compensate the negative effects of slow heating rates. However, there are situations in which slow heating rates closely mimic natural conditions, as those encountered by some aquatic ectotherms. These heating rates may be an issue of major concern in these species, given its negative impact on CT max and its adaptive potential.Key-words: ectotherms, energy expenditure, evolvability, heritability, phenotypic plasticity, thermal ramping, thermal toleranceThe reason why plants and animals (including men) are so sensitive to temperature is that they are chemical machines …. Many of them do burn even at ordinary temperatures, but so slowly that we don't notice any great change even within a lifetime. (Haldane 1940, pp. 69-70)

Summary1. An increasing body of knowledge suggests that the estimation of critical upper thermal limits (CT max ) is highly dependent on the experimental methodology employed. Here, we employ a theoretical approach to analyse how estimates of CT max (knock-down temperatures and times) are affected by measurement protocol. 2. Our model is able to reproduce the results of empirical studies on Drosophila melanogaster, suggesting that it adequately mimics organismal responses during assays. With simulated data sets, we also show that many experimental protocols result in unreliable and often highly biased estimates of CT max in Drosophila and possibly in other ectotherms. 3. The confounding effects of stochasticity, resource depletion (or fatigue) and short-term acclimatory responses are expected to be higher in longer assays, and therefore, short assays should be generally preferred. The experimental protocol of choice must also take into consideration the range in which measurement accuracy is not affected and the potential problems of thermal inertia in larger organisms. 4. Our findings justify previous concerns that the methodology may have a greater impact on estimates of CT max than the biological process under study, and explain why many studies on the subject have often reported inconsistent and even contradictory results.

Polymorphism at the ADH2 and ADH3 loci of alcohol dehydrogenase (ADH) has been shown to have an effect on the predisposition to alcoholism in Asian individuals. However, the results are not conclusive for white individuals. We have analyzed the ADH genotype of 876 white individuals from Spain (n ‫؍‬ 251), France (n ‫؍‬ 160), Germany (n ‫؍‬ 184), Sweden (n ‫؍‬ 88), and Poland (n ‫؍‬ 193). Peripheral blood samples from healthy controls and groups of patients with viral cirrhosis and alcohol-induced cirrhosis, as well as alcoholics with no liver disease, were collected on filter paper. Genotyping of the ADH2 and ADH3 loci was performed using polymerase chain reactionrestriction fragment length polymorphism methods on white cell DNA. In healthy controls, ADH2*2 frequencies ranged from 0% (France) to 5.4% (Spain), whereas ADH3*1 frequencies ranged from 47.6% (Germany) to 62.5% (Sweden). Statistically significant differences were not found, however, between controls from different countries, nor between patients with alcoholism and/or liver disease. When all individuals were grouped in nonalcoholics (n ‫؍‬ 451) and alcoholics (n ‫؍‬ 425), ADH2*2 frequency was higher in nonalcoholics (3.8%) than in alcoholics (1.3%) (P ‫؍‬ .0016), whereas the ADH3 alleles did not show differences. Linkage disequilibrium was found between ADH2 and ADH3, resulting in an association of the alleles ADH2*2 and ADH3*1, both coding for the most active enzymatic forms. In conclusion, the ADH2*2 allele decreases the risk for alcoholism, whereas the ADH2*2 and ADH3*1 alleles are found to be associated in the European population. (HEPATOLOGY 2000;31:984-989.)Ingested alcohol is mostly metabolized in the liver by the successive action of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). Both enzymes exhibit genetic polymorphisms that influence the rate of conversion of ethanol to acetaldehyde, and of acetaldehyde to acetate. It has been consistently reported that ALDH2 is the most important alcohol-metabolizing gene affecting predisposition to alcoholism in Asian populations. The prevalence of the ALDH2*2 allele, which codes for a physiologically inactive mitochondrial ALDH form, is lower in alcoholics than in nonalcoholics. [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] However, this allele has not been found in white individuals. 21 Regarding ADH, polymorphism is detected at the ADH2 and ADH3 loci. Alleles of ADH2 found in whites and Asians are ADH2*1 and ADH2*2, which encode for the low activity (␤1) and high activity (␤2) subunits, respectively. The kcat values for the resulting dimeric isozymes are very different: 9.2 min Ϫ1 for ␤1␤1 and 400 min Ϫ1 for ␤2␤2. 22 The ADH2*2 frequency is much higher in Asians (60%-80%) than in whites (0%-10%). 21 ADH3 alleles are ADH3*1 and ADH3*2, which produce the ␥1 and ␥2 subunits. The ␥1␥1 isozyme (kcat ϭ 87 min Ϫ1 ) is moderately more active than the ␥2␥2 isozyme (kcat ϭ 35 min Ϫ1 ). 22 ADH3*1 frequency is about 50% to 60% in whites and higher than 90% in Asians. 3,23 ...