Environmental and genetic control of cold tolerance in the Glanville fritillary butterfly.

Thermal tolerance has a major effect on individual fitness and species distributions and can be determined by genetic variation and phenotypic plasticity. We investigate the effects of developmental and adult thermal conditions on cold tolerance, measured as chill coma recovery (CCR) time, during the early and late adult stage in the Glanville fritillary butterfly. We also investigate the genetic basis of cold tolerance by associating CCR variation with polymorphisms in candidate genes that have a known role in insect physiology. Our results demonstrate that a cooler developmental temperature leads to reduced cold tolerance in the early adult stage, whereas cooler conditions during the adult stage lead to increased cold tolerance. This suggests that adult acclimation, but not developmental plasticity, of adult cold tolerance is adaptive. This could be explained by the ecological conditions the Glanville fritillary experiences in the field, where temperature during early summer, but not spring, is predictive of thermal conditions during the butterfly's flight season. In addition, an amino acid polymorphism (Ala-Glu) in the gene flightin, which has a known function in insect flight and locomotion, was associated with CCR. These amino acids have distinct biochemical properties and may thus affect protein function and/or structure. To our knowledge, our study is the first to link genetic variation in flightin to cold tolerance, or thermal adaptation in general.

Fight or flight? - Flight increases immune gene expression but does not help to fight an infection.

Flight represents a key trait in most insects, being energetically extremely demanding, yet often necessary for foraging and reproduction. Additionally, dispersal via flight is especially important for species living in fragmented landscapes. Even though, based on life-history theory, a negative relationship may be expected between flight and immunity, a number of previous studies have indicated flight to induce an increased immune response. In this study, we assessed whether induced immunity (i.e. immune gene expression) in response to 15-min forced flight treatment impacts individual survival of bacterial infection in the Glanville fritillary butterfly (Melitaea cinxia). We were able to confirm previous findings of flight-induced immune gene expression, but still observed substantially stronger effects on both gene expression levels and life span due to bacterial infection compared to flight treatment. Even though gene expression levels of some immunity-related genes were elevated due to flight, these individuals did not show increased survival of bacterial infection, indicating that flight-induced immune activation does not completely protect them from the negative effects of bacterial infection. Finally, an interaction between flight and immune treatment indicated a potential trade-off: flight treatment increased immune gene expression in naïve individuals only, whereas in infected individuals no increase in immune gene expression was induced by flight. Our results suggest that the up-regulation of immune genes upon flight is based on a general stress response rather than reflecting an adaptive response to cope with potential infections during flight or in new habitats.

Temperature- and sex-related effects of serine protease alleles on larval development in the Glanville fritillary butterfly.

The body reserves of adult Lepidoptera are accumulated during larval development. In the Glanville fritillary butterfly, larger body size increases female fecundity, but in males fast larval development and early eclosion, rather than large body size, increase mating success and hence fitness. Larval growth rate is highly heritable, but genetic variation associated with larval development is largely unknown. By comparing the Glanville fritillary population living in the Åland Islands in northern Europe with a population in Nantaizi in China, within the source of the post-glacial range expansion, we identified candidate genes with reduced variation in Åland, potentially affected by selection under cooler climatic conditions than in Nantaizi. We conducted an association study of larval growth traits by genotyping the extremes of phenotypic trait distributions for 23 SNPs in 10 genes. Three genes in clip-domain serine protease family were associated with larval growth rate, development time and pupal weight. Additive effects of two SNPs in the prophenoloxidase-activating proteinase-3 (ProPO3) gene, related to melanization, showed elevated growth rate in high temperature but reduced growth rate in moderate temperature. The allelic effects of the vitellin-degrading protease precursor gene on development time were opposite in the two sexes, one genotype being associated with long development time and heavy larvae in females but short development time in males. Sexually antagonistic selection is here evident in spite of sexual size dimorphism.

Direct and trans-generational responses to food deprivation during development in the Glanville fritillary butterfly.

Life history characteristics and resulting fitness consequences manifest not only in an individual experiencing environmental conditions but also in its offspring via trans-generational effects. We conducted a set of experiments to assess the direct and trans-generational effects of food deprivation in the Glanville fritillary butterfly Melitaea cinxia. Food availability was manipulated during the final stages of larval development and performance was assessed during two generations. Direct responses to food deprivation were relatively minor. Food-deprived individuals compensated, via increased development time, to reach a similar mass as adults from the control group. Delayed costs of compensatory growth were observed, as food-deprived individuals had either reduced fecundity or lifespan depending on the type of feeding treatment they had experienced (intermittent vs. continuous). Female food deprivation did not directly affect her offspring's developmental trajectory, but the way the offspring coped with food deprivation. Offspring of mothers from control or intermittent starvation treatments reached the size of those in the control group via increased development time when being starved. In contrast, offspring of mothers that had experienced 2 days of continuous food deprivation grew even larger than control animals, when deprived of food themselves. Offspring of food-deprived Glanville fritillary initially showed poor immune response to parasitism, but not later on in development.

Heritability of dispersal rate and other life history traits in the Glanville fritillary butterfly.

Knowing the variances and heritabilities (h(2)) of life history traits in populations living under natural conditions is necessary for a mechanistic understanding of respective evolutionary processes. I estimated heritabilities of several life history traits, including dispersal rate, body mass, age at first reproduction, egg mass, clutch size and lifetime reproductive success, in the Glanville fritillary butterfly (Melitaea cinxia) using parent-offspring regression. Experiments were conducted under field conditions in a large population cage (32 x 26 m). Heritability estimates ranged from zero to almost one and several were significantly different from zero. Body size for both sexes, female age at first reproduction and egg weight were all moderately to highly heritable, whereas heritabilities were low or non-existent in clutch size and lifetime egg production. Heritability estimates for dispersal rate varied between the sexes, so that dispersal was heritable from mother to her female offspring only. This finding is consistent with previous results showing that the F1 female but not male offspring of females that naturally established new populations in the field are significantly more dispersive than butterflies in old populations.

Effect of linseed cake supplementation on digestibility and faecal and haematological parameters in dogs.

The experiment was conducted with a 6 x 4 cyclic changeover design. Six adult Alaskan huskies were assigned to diets containing 0%, 2%, 4%, 6% and 8% linseed cake. A linear decrease in the apparent digestibilities of organic matter (OM) (p < 0.020), ether extract (EE) (p < 0.003), neutral detergent fibre (p < 0.015), nitrogen-free extract (NFE) (p < 0.001) and crude carbohydrates (p < 0.001) was noted. Digestibilities of dry matter (DM) and crude protein (CP) were unaffected by the treatment. Wet weight of faeces increased (p < 0.001) and DM of faeces decreased (p < 0.001) linearly as the percentage of dietary linseed cake increased. Serum cholesterol concentration tended to decrease (p = 0.094) as the proportion of linseed cake increased. Apparent OM, EE, CP and NFE digestibility coefficients for linseed cake were 52%, 77%, 54% and 37%, respectively, and the digestible energy (DE) value was 13.22 MJ/kg DM. Our results showed that working and racing dogs fed the extruded and kibbled diet can utilize up to 4.2% linseed cake of diet DM as a fibre source without severe reductions in nutrient digestibility or feed consumption. Even higher levels of linseed cake, up to 8.5% of diet DM, can be used for normal or obese dogs, but more faeces will be excreted.

Very acute cardiac toxicity during BEAC chemotherapy in non-Hodgkin's lymphoma patients undergoing autologous stem cell transplantation.

Cardiotoxicity is potentially the most threatening nonhaematological side effect of high-dose CY. We prospectively evaluated the very acute cardiac effects of high-dose CY in 17 adult non-Hodgkin's lymphoma (NHL) patients receiving CY 1500 mg/m2/day as a part of BEAC high-dose therapy (HDT). Magnetic resonance imaging (MRI) and plasma natriuretic peptide (NT-proBNP, NT-proANP) measurements were performed prior to HDT (d-7) and just after completing HDT (d-2). After the high-dose CY left atrial end-systolic area increased from 15.2+/-1.2 to 18.5+/-1.4 cm2 (P=0.001), left ventricular end-diastolic volume from 136.1+/-12.3 to 156.6+/-11.1 cm3 (P=0.04) and left ventricular end-systolic volume from 67.4+/-7.8 to 75.3+/-7.1 cm3 (P=0.018). However, no significant change in left ventricular ejection fraction (LVEF) was observed. At the same time, plasma levels of NT-proBNP increased from 134.9+/-53.3 to 547.1+/-168.4 pmol/l (P=0.003) and NT-proANP from 481.1+/-105.5 to 1056.6+/-193.1 pmol/l (P=0.001), respectively. To conclude, high-dose CY results in very acute cardiac toxicity characterised by enlargement of the heart chambers in NHL patients previously treated with anthracyclines. This toxicity can be detected with increased concentrations of circulating natriuretic peptides but not with LVEF measurement.

Extrastriatal dopamine D 2/3 receptor density and distribution in drug-naive schizophrenic patients.

Several lines of studies have suggested the importance of cortical dopamine (DA) transmission in the pathophysiology of schizophrenia. The putative alteration of striatal D(2) receptor density in schizophrenia has been studied intensely, although extrastriatal DA activity may be more relevant for behavioral symptoms. The aim of this study was to explore extrastriatal D(2/3) density in drug-naive schizophrenic patients. We studied the extrastriatal D(2/3) receptor binding with a novel high-affinity single-photon emission tomography ligand epidepride in seven drug-naive schizophrenic patients and seven matched controls. The symptoms were rated with Positive and Negative Syndrome Scale for Schizophrenia. The findings indicated an extremely low D(2/3) receptor binding among patients in temporal cortex in both hemispheres when compared with controls (effect size 2.0-2.3), and the D(2/3) levels had negative correlations with general psychopathological (r from -0.86 to -0.90) and negative (r from -0.37 to -0.55) schizophrenic symptoms. These results support the previous hypothesis on dysfunction of mesocortical DA function behind the cognitive and negative symptoms in schizophrenia.

PET imaging using a triple-head gamma camera.

Interest in clinical fluorodeoxyglucose (FDG) imaging with multiple-head gamma cameras is growing. To improve sensitivity, triple-head coincidence imaging has been proposed. We report our initial experiences with a triple-head coincidence gamma camera with 19 mm sodium iodide crystal thickness. Several positron emission tomography-image quality parameters were evaluated using a Carlson and line source phantom. The system sensitivity with two-dimensional axial shields was 830 cps kBq-1 ml-1 and maximum noise equivalent count rate 1900 cps for an 18F-activity of 50 MBq. The imaging resolution was in central axial 7.0 mm and in central transaxial 7.6 mm, respectively. The average scatter fraction in scattered media was 29%. Clinical brain, heart and whole body images studies with [18F]FDG were acquired and they show good correlation with the phantom image quality. As a conclusion, triple-head coincidence gamma camera provides relatively high-count rate imaging with good contrast and resolution.

SPECT and neuropsychological performance in severe depression treated with ECT.

BACKGROUND: In severe depression, studies of regional cerebral blood flow (rCBF) by SPECT have not produced uniform results. The association between changes in SPECT and electroconvulsive therapy (ECT) has shown somewhat conflicting data. No data are available on benzodiazepine receptor function SPECT studies in ECT. METHODS: Twenty drug-resistant adult inpatients fulfilling the DSM-IIIR criteria for major depression were studied by SPECT (rCBF by relative ECD uptake in all, and benzodiazepine receptor function by iomazenil uptake in five subjects) before and 1 week after clinically successful bitemporal ECT. Clinical and neuropsychological test scores were used as references for the possible changes in SPECT. RESULTS: An increased perfusion after ECT was observed in right temporal and bilateral parietal cortices, whereas no reductions in relative ECD uptake were seen after ECT. Iomazenil-SPECT revealed a highly significant increase in the benzodiazepine receptor uptake in all studied cortical regions except temporal cortices. CONCLUSIONS: Clinically successful ECT was associated with changes in vascular perfusion and GABAergic neurotransmission, providing new evidence for the mechanism of action of ECT and for the neurobiology of severe drug-resistant depression.

Extension of anther culture to several genotypes of cultivated oats.

To improve plant regeneration from oat anther culture, the basic medium, hormonal supplements and genotype effect were studied. Six of the 14 genotypes tested regenerated plants. Cultivars Kolbu, Katri, Stout and naked oat Lisbeth produced green plants, cultivars Virma and line OT 257 only albinos. The total number of green plantlets regenerated was 22, of which 13 (11 haploid, 2 doubled haploid) survived into the greenhouse, and 37 albinos. Regenerable-type embryos were induced from heat-pretreated anthers on media containing 2, 3 or 5 mg l-1 2,4-dichlorophenoxyacetic acid and 0.2 or 0.5 mg l-1 kinetin as hormonal supplements. 6-Benzylaminopurine promoted albino plant regeneration especially in W14 medium. Colchicine treatment was applied successfully to haploid regenerants.

Striatal dopamine transporter density in major depression.

RATIONALE: There are no previous data available regarding [123I]beta-CIT binding to the dopamine transporter sites in the basal ganglia in depressed patients. OBJECTIVE: The present study tested the hypothesis that the brain DAT density in depressed patients is lower than that in matched healthy controls. METHODS: Fifteen drug-naive outpatients with major depression and 18 healthy controls were investigated using single photon emission computerized tomography (SPECT) with a high-affinity dopamine transporter specific radioligand. 123I-labeled beta-CIT (2beta-carbomethoxy-3beta-(4-iodophenyl-tropane). RESULTS: We found a significantly higher [123I]beta-CIT uptake in both sides of the basal ganglia in patients with major depression than in the controls (Mann-Whitney U-test, P = 0.002 on the right and P = 0.003 on the left). CONCLUSIONS: The radioligand uptake reflecting the DAT density was significantly higher among the patients than in the controls. This finding is unexpected, since it is generally believed that monoaminergic neurotransmission is lower in depression, and therefore it could be assumed that a reduction in dopamine transmission would lead to secondary down-regulation of DAT density. However, it is possible that up-regulation of the DAT may be the primary alteration, which leads to lower intrasynaptic dopamine concentration and to lower dopamine neural transmission.

Effect of a post exercise fat-supplemented diet on muscle glycogen repletion.

Low muscle glycogen at the beginning of exercise may adversely affect performance, increase protein degradation and contribute to the onset of fatigue. As horses are sometimes required to compete on consecutive days both in racing and endurance types of competition, optimal muscle glycogen repletion may improve performance on the day following a race day. The purpose of this experiment was to study the effects of fat supplementation on repletion of muscle glycogen. Twelve Finnhorses performed an exercise test on a treadmill, and 2 and 4 h later they received hay and concentrate (Trial A). Two weeks later these horses performed the same exercise test and were fed the same diet supplemented with either 1000 g of carbohydrate or 400 g of vegetable oil (Trial B). A third trial (Trial C) was 3 weeks later, identical to Trial B, except that the fat group had already been adapted to dietary fat for 3 weeks. Blood samples were analysed for lactate, glucose, glycerol, triglycerides, NEFA, cholesterol, beta-OH-butyrate, insulin and glucagon and muscle samples were analysed for glycogen and triglycerides. The results indicate that in horses not adapted to fat feeding, fat supplementation slows the rate of muscle glycogen repletion, and that after an adaptation period, fat supplementation does not alter the rate of muscle glycogen repletion compared to the rate with a normal diet.

Restoration of water and electrolyte balance in horses after repeated exercise in hot and humid conditions.

Nine adult riding horses performed on a treadmill a competition exercise test (CET) and 24 h later a standardised exercise test (SET) at mean temperature 28 degrees C and relative humidity 58%. Each horse performed the tests 5 times at 2 week intervals. The horses were given isotonic glucose-electrolyte solution via a nasogastric tube 30 min after the CET, except after the last trial when water was given. Blood lactate, plasma concentration of aldosterone, arginine vasopressin, protein, sodium, potassium, chloride, magnesium and calcium were measured. Heart rate at blood lactate concentration 4 mmol/l (PLa4) in the SET, plasma volume (PV) and red cell volume (CV) were determined. Mean weight loss after CET was 3.1% and did not change significantly during the study. Bodyweight loss (BWL2) before SET (-1.8% after the first trial) decreased linearly with time (P < 0.05) and was -0.8% after the fourth trial. After the fifth trial, when only water was given, the weight loss was -2.5%. PLa4 correlated significantly (P < 0.001) with BWL2 when BWL2 was greater than -1.2%. The horses acclimated to exercise in hot and humid conditions as indicated by better recovery of bodyweight, increased PV, lower peak lactate concentrations and heart rate and attenuated decrease in the concentrations of sodium and chloride. It is concluded that changes in bodyweight provide a good indication of recovery of horses after exercise in hot and humid conditions; and administration of an isotonic glucose-electrolyte rehydration solution after exercise helps to overcome dehydration better than water alone.

Health habits in a Finnish village.

This study is part of a collaborative study entitled "Life-style and health" which is being conducted in Finland, India and Tanzania, using the same methods. The aim of the study is to describe health habits (smoking, alcohol consumption, use of medical drugs, exposure to sugars and hygiene habits) in Viekijärvi, a village in eastern Finland. Of the villagers 50% of those aged 20 years and over were randomly selected to take part in the study. Of these, 72% (n = 179) returned the standard questionnaire they had been sent. Smoking was as common among men in Viekijärvi as in Finland in general, but less common among women. Alcohol consumption reflected the national consumption. The use of medical drugs was high: 45% of respondents used such preparations daily. The most common exposure to sugar on a daily basis was sugar-sweetened coffee. Most people in Viekijärvi (89%) brushed their teeth at least once a day. A strong positive correlation was found between frequencies of toothbrushing and handwashing mirroring unidimensional hygienic behaviour. The results of this study will form a basis for a health promotion programme, to be developed by the villagers themselves.