Serum 25-hydroxyvitamin D is associated with major cardiovascular risk factors and cardiac structure and function in patients with coronary artery disease.

BACKGROUND AND AIMS: Vitamin D deficiency has been associated with increased risk for cardiovascular (CV) disease, but the possible effects of Vitamin D on cardiac structure and function are not well characterized. METHODS AND RESULTS: The correlation between 25-hydroxyvitamin D levels and metabolic and cardiac echocardiographic parameters was studied in ARTEMIS study population including 831diabetic and 659 non-diabetic patients with stable coronary artery disease (CAD). Low levels of Vitamin D were associated with high BMI (p < 0.001), high total and LDL cholesterol and triglyceride levels (p < 0.001 for all) in both diabetics and non-diabetics. Among non-diabetic patients, low Vitamin D was also associated independently with elevated systolic and diastolic blood pressure (p < 0.005). Low Vitamin D levels were independently associated with reduced left ventricular (LV) ejection fraction (p < 0.005) and increased left atrial diameter (p < 0.03) measured by cardiac ultrasound by 2-dimensional echo. In the non-diabetic group, low Vitamin D levels were associated with impaired LV filling (high E/E') (p < 0.03) and low E/A mitral flow pattern measured by Doppler echocardiography (p < 0.05). Among diabetics, low Vitamin D levels were also related to increased LV end-systolic diameter (p < 0.05) and right ventricular diameter (p < 0.005). The association between LV diastolic filling (E/E') and Vitamin D levels was significant (p < 0.01) after adjustment for the commonly recognized risk factors of diastolic dysfunction in linear regression analysis. CONCLUSIONS: Low Vitamin D is associated with several major cardiovascular risk factors and cardiac structural changes including impaired systolic and diastolic function, which together may explain the association of low Vitamin D to worse cardiovascular outcome.

Beyond MHC: signals of elevated selection pressure on Atlantic salmon (Salmo salar) immune-relevant loci.

Using Atlantic salmon (Salmo salar) as a model system, we investigated whether 18 microsatellites tightly linked to immune-relevant genes have experienced different selection pressures than 76 loci with no obvious association with immune function. Immune-relevant loci were identified as outliers by two outlier tests significantly more often than nonimmune linked loci (22% vs. 1.6%). In addition, the allele frequencies of immune relevant markers were more often correlated with latitude and temperature. Combined, these results support the hypothesis that immune-relevant loci more frequently exhibit footprints of selection than other loci. They also indicate that the correlation between immune-relevant loci and latitude may be due to temperature-induced differences in pathogen-driven selection or some other environmental factor correlated with latitude.

High Gyrodactylus salaris infection rate in triploid Atlantic salmon Salmo salar.

We describe an unusually high infection rate of Gyrodactylus salaris Malmberg in juvenile Atlantic salmon Salmo salar L. of Baltic Sea origin, which are generally believed to be more resistant to G. salaris than East Atlantic salmon populations. Based on analyses of mitochondrial (complete cytochrome oxidase 1 [CO1] gene, 1548 bp) and nuclear (ADNAM1, 435 bp; internal transcribed spacer [ITS] rDNA region, 1232 bp) DNA fragments, the closest relatives of the characterized Estonian G. salaris strain were parasites found off the Swedish west coast and in Raasakka hatchery, Iijoki (Baltic Sea, Finland). Analyses of 14 microsatellite loci of the host S. salarrevealed that approximately 40% of studied fish were triploids. We subsequently identified triploid Atlantic salmon of Baltic origin as more susceptible to G. salaris infection than their diploid counterparts, possibly due to compromised complement-dependent immune pathways in triploid salmon. This is in accordance with earlier studies that have shown elevated susceptibility of triploids to various viral or bacterial pathogens, and represents one of the first reports of increased susceptibility of triploid salmonid fish to an ectoparasite. However, further experimental work is needed to determine whether triploid Atlantic salmon is generally more susceptible to G. salaris compared to their diploid counterparts, irrespective of the particular triploidization method and population of origin.

Use of differential expression data for identification of novel immune relevant expressed sequence tag-linked microsatellite markers in Atlantic salmon (Salmo salar L.).

Despite the large number of genes contributing to the immune response, wildlife immunogenetic studies have tended to focus mostly on the major histocompatibility complex-related genes. Here, we utilized previously published microarray and competitive RNA hybridization information to identify 3750 immune relevant Atlantic salmon (Salmo salar) expressed sequence tags. We then identified those expressed sequence tags containing microsatellites and subsequently designed 48 primer pairs and tested them for polymorphism in Atlantic salmon. Altogether, 16 polymorphic markers were characterized, with allele numbers ranging from two to 18, and these 16 loci were further tested in five other salmonid species.

Hybrid origin of Baltic salmon-specific parasite Gyrodactylus salaris: a model for speciation by host switch for hemiclonal organisms.

Host switching explains the high species number of ectoparasitic, viviparous, mainly parthenogenetic but potentially hermaphroditic flatworms of the genus Gyrodactylus. The starlike mitochondrial phylogeny of Gyrodactylus salaris suggested parallel divergence of several clades on grayling (also named as Gyrodactylus thymalli) and an embedded sister clade on Baltic salmon. The hypothesis that the parasite switched from grayling to salmon during the glacial diaspora was tested using a 493-bp nuclear DNA marker ADNAM1. The parasites on salmon in lakes Onega and Ladoga were heterozygous for divergent ADNAM1 alleles WS1 and BS1, found as nearly fixed in grayling parasites in the White Sea and Baltic Sea basins, respectively. In the Baltic salmon-specific mtDNA clade, the WS/BS heterozygosity was maintained in 23 out of the 24 local clones. The permanently heterozygous clade was endemic in the Baltic Sea basin, and it had accumulated variation in mtDNA (31 variable sites on 1600 bp) and in the alleles of the nuclear locus (two point mutations and three nucleotide conversions along 493 bp). Mendelian shuffling of the nuclear alleles between the local clones indicated rare sex within the clade, but the WS/BS heterozygosity was lost in only one salmon hatchery clone, which was heterozygous WS1/WS3. The Baltic salmon-specific G. salaris lineage was monophyletic, descending from a single historical hybridization and consequential host switch, frozen by permanent heterozygosity. A possible time for the hybridization of grayling parasite strains from the White Sea and Baltic Sea basins was during the Eemian interglacial 132 000 years bp. Strains having a separate divergent mtDNA observed on farmed rainbow trout, and on salmon in Russian lake Kuito were suggested to be clones derived from secondary and tertiary recombination events.

Deep divergence among subgenera of Gyrodactylus inferred from rDNA ITS region.

This paper adds new insight to a molecular phylogeny of Gyrodactylus, based on a complete sequence of the ITS rDNA region of 4 subgenera and a more detailed molecular analysis. We propose a hierarchical approach in elucidating the phylogeny of this species-rich genus. A total of 37 sequences (915-1239 bp) from 10 representative species from 4 out of 6 subgenera, as defined by Malmberg (1970), are included in the analysis. Genetic differences observed at the 5.8S locus provide objective criteria to separate (sub)genera, while deep genetic differences of the spacers form a sound basis for species-specific identification. We demonstrate that each Gyrodactylus subgenus possesses a unique sequence of the 5.8S gene. Thus, there is concordance between the 5.8S gene and the excretory system used by Malmberg (1970) as a diagnostic character of subgenus status. At the species level, there is a discrepancy between morphological and molecular variation. Whereas the morphological variation, expressed in the shape and size of the attachment apparatus, is very low, the molecular variation, expressed at the I

Matrilinear phylogeography of Atlantic salmon (Salmo salar L.) in Europe and postglacial colonization of the Baltic Sea area.

Sixty-four samples from 46 salmon populations totalling 2369 specimens were used for polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mitochondrial ND1 region. The final analyses included 3095 specimens from 60 populations in Northern Europe. A subsample was analysed by RFLP of ND3/4/5/6. Representative RFLP haplotypes from different parts of the distribution area were sequenced and the phylogeny of European haplotypes and their relations to the North American lineage was described. The four common European haplotypes derive from the ancestral ND1-BBBA (rooting the European clade to the North American) by one-step substitutions: AAAA < AABA < BBBA > BBBB. The Swedish west-coast populations differ from the geographically close southern Baltic, indicating absence of inward and limited outward gene flow through the Danish straits during the last 8000 years. Within the Baltic Sea, only three ND1 haplotypes were detected and there was no variation for ND3/4/5/6. In the whole southern Baltic and in lakes Vänern, Ladoga and Onega the haplotype AABA dominated. Proposed postglacial colonization routes to the Baltic Sea are discussed in relation to the haplotype distribution pattern.

Localization of genes affecting species differences in male courtship song between Drosophila virilis and D. littoralis.

The males of six species of the Drosophila virilis group (including D. virilis) keep their wings extended while producing a train of sound pulses, where the pulses follow each other without any pause. The males of the remaining five species of the group produce only one sound pulse during each wing extension/vibration, which results in species-specific songs with long pauses (in D. littoralis about 300 ms) between successive sound pulses. Genetic analyses of the differences between the songs of D. virilis and D. littoralis showed that species-specific song traits are affected by genes on the X chromosome, and for the length of pause, also by genes on chromosomes 3 and 4. The X chromosomal genes having a major impact on pulse and pause length were tightly linked with white, apricot and notched marker genes located at the proximal third of the chromosome. A large inversion in D. littoralis, marked by notched, prevents more precise localization of these genes by classical crossing methods.

The colonization history and present-day population structure of the european great tit (Parus major major)

The colonization history and present-day population structure of the European subspecies of the great tit Parus major major were studied using mitochondrial control region sequences. One major haplotype was found in all but one of the eight sampled populations from Spain to northern Finland. The other haplotypes differed from the common one by just a few substitutions; the overall nucleotide diversity was 0.00187 and haplotype diversity 0.8633. No population structuring was detected. The mismatch distribution followed the expected distribution of an expanding population. The estimated time to the most recent common ancestor coincides with the last glacial period. The results suggest that P. m. major survived the last glacial period in a single isolated refuge probably by the Mediterranean Sea. This was followed by rapid colonization of the European continent and population growth. The most recent range expansion northwards is still occurring. Gene flow between the sampled populations is extensive. It is aided by juvenile dispersal, long-distance movements of juvenile flocks and partial migration in the northern parts of the great tit's range.

Plasma vasoactive peptides after acute myocardial infarction in relation to left ventricular dysfunction.

We measured plasma concentrations of vasoactive peptides in 32 patients with acute myocardial infarction and evaluated their value as markers of left ventricular dysfunction. Plasma levels of atrial natriuretic peptide (ANP), the N-terminal fragment of proANP (NT-proANP), B-type natriuretic peptide (BNP) and endothelin-1 were measured serially by radioimmunoassays. The infarct size was estimated from the creatine kinase MB release curve. Coronary angiography and left ventricular cineangiography were performed in all patients during hospitalization and 6 months later in 15 patients. Myocardial infarction caused an increase in vasoactive peptides, the highest values for ANP (36.5+/-6.79 pmol/l), NT-proANP (1130+/-170 pmol/l) and endothelin-1 (9.72+/-0.68 pmol/l) being found on admission and those for BNP (56.0+/-7.13 pmol/l) on Day 2. Plasma levels of natriuretic peptides were dependent on infarct size, its location and degree of myocardial dysfunction and that of BNP also on infarct artery patency. Plasma endothelin-1 level was higher in patients with TIMI 3 than TIMI 0-2 flow. Plasma vasoactive peptides remained elevated during the 6-month follow-up period and they were dependent on the degree of myocardial dysfunction. BNP measured on any day of hospitalization showed the best correlation with ejection fraction measured during the acute phase of infarction or at 6 months. The results show that BNP is the best indicator of left ventricular dysfunction after myocardial infarction and its reliability is not dependent on the time point of measurement.

Mitochondrial control region polymorphism reveal high amount of gene flow in Fennoscandian willow tits (Parus montanus borealis).

We studied the genetic variability and differentiation of two Fennoscandian willow tit (Parus montanus borealis) populations located ca. 1000 km apart in Finland and Sweden by using the control region sequences of the mitochondrial DNA. Individual variation in the control region was extensive since all the 13 Finnish and the 12 Swedish individuals had unique haplotypes and the mean pairwise genetic distance resulted in 0.0052 (range 0.0008-0.0109). In the minimum spanning network connecting the genotypes, the two populations were completely intermingled. The mismatch distribution of the combined data set was very close to expected distribution of an expanding population. This result was supported by a significantly negative Tajima's D value. The sequence data indicate that (1) the long-term effective population size of the breeding willow tits has been large (122,000 and 110,000 females for the Finnish and the Swedish populations, respectively); and that (2) the gene exchange between distant localities is/has been extensive. Dispersal area for the Finnish females was estimated to be about 19,000-30,000 km2 and for the Swedish, 22,000-28,000 km2. Thus, the whole Fennoscandian population can be regarded as one panmictic unit, without any subdivisions to local demes. The amount of gene flow is remarkable because the willow tit has been considered a highly sedentary species. Provided that adult birds are site-tenacious, the gene flow must occur through juvenile summer dispersal, or irruptive autumn invasions, or both.

The Y chromosomes of Drosophila lummei and D. novamexicana differ in fertility factors.

Many first-generation interspecific hybrids between the five species of the Drosophila virilis subgroup are fertile. Fertile hybrids can be utilized in the analysis of sterile hybrids, by creating new combinations of genetic elements to identify cross-incompatible chromosomes or chromosome segments. Here, crosses starting from D. lummei, D. virilis, D. novamexicana and D. americana were produced to analyse the 96.4% male sterility in the progeny of D. virilis females mated with D. novamexicana males (sterility based on sperm motility). In autosomally identical heterozygous combination, the substitution of a large segment of the D. virilis X chromosome with a double or a triple inversion from the sister taxa decreased male sterility from 96.4% (Xvirilis) to 11% (Xvirilis/novamexicana), 19% (Xvirilis/americana) and 34% (Xvirilis/lummei). This result shows that the Xvirilis interaction with D. novamexicana elements (Y and/or autosomes) is a major component in the F1 sterility observed. When males with a substituted Ylummei in otherwise pure D. novamexicana were crossed with D. virilis females, all the F1 males were fertile. We constructed combinations of intact Xvirilis, recombinant Xvirilis/novamexicana, Ynovamexicana and Ylummei chromosomes in homozygous D. virilis and heterozygous D. virilis/D. novamexicana autosomal backgrounds. In such combinations, the Y chromosome interacted strongly not only with the nonconspecific X chromosome, but with several autosomes. Autosomal hybrid sterility contributions tend to be recessive and not present in the F1. In this species group, the role of the Y chromosome is central in contributing to the phenomenon called Haldane's rule of sterility, but the Y chromosomal incompatibility factor of D. novamexicana is the most divergent one.

Collagen scar formation after acute myocardial infarction: relationships to infarct size, left ventricular function, and coronary artery patency.

BACKGROUND: Left ventricular function after acute myocardial infarction (AMI) is determined by the expansion of the infarct zone and remodeling of the noninfarcted myocardium. An occluded infarct-related artery (IRA) is an independent risk factor for remodeling. METHODS AND RESULTS: Changes in myocardial collagen metabolism were evaluated in 36 patients with suspected AMI. The plasma creatine kinase MB fraction and myoglobin release curves were analyzed for assessment of early reperfusion and infarct size. Collagen scar formation was evaluated by measurement of serum concentrations of the aminoterminal propeptide of type III procollagen (PIIINP), the aminoterminal propeptide of type I procollagen (intact PINP), and the carboxyterminal propeptide of type I procollagen (PICP). Plasma renin activity and urine excretion of cortisol and aldosterone were also measured. Coronary angiography and left ventricular cineangiography were performed during early hospitalization. The serum concentration of PIIINP increased from 3.50+/-0.20 to a maximum of 5.08+/-0.36 microg/L (n=32) in the patients with AMI, whereas the concentrations of intact PINP and PICP tended to decrease. The area under the curve (AUC) of PIIINP during the first 10 postinfarction days was larger in patients with severe heart failure or ejection fractions < or = 40% than in those with no heart failure or with an ejection fraction > 40% (P<.05 and P<.01, respectively), and it was also larger in the patients with TIMI grade 0 to 2 flows than in those with TIMI 3 flows (P<.05), despite similar enzymatically determined infarct sizes. No significant correlations between PIIINP and neurohumoral parameters were observed. The AUC of PIIINP and the change in PIIINP during the first 4 days were significantly correlated with indices of cardiac function. CONCLUSIONS: Collagen scar formation after AMI can be quantified by measurement of serum PIIINP concentrations. Scar formation is more prominent in large infarctions causing left ventricular dysfunction and in patients with occluded IRAs.

Relation between low dose of hydrochlorothiazide, antihypertensive effect and adverse effects.

Thiazide diuretics are widely used in the drug treatment of hypertension but their dose-response curves for the antihypertensive and adverse metabolic effects differ. To characterize the lower end of the dose-response curve a double-blind, parallel group trial was performed as multicentre study in Scandinavia. One hundred and eleven patients with newly diagnosed or previously treated mild to moderate hypertension (untreated diastolic blood pressure of 95-115 mmHg after 4 weeks placebo) were randomly allocated to various doses of hydrochlorothiazide (3, 6, 12.5 or 25 mg) or placebo for 6 weeks. Blood pressure and biochemical variables (plasma renin activity, serum potassium, magnesium, urate, fasting glucose, total cholesterol, HDL-cholesterol, triglycerides and apolipoproteins A1 and B were measured. 12.5 mg hydrochlorothiazide had a borderline effect on blood pressure whilst 25 mg had a definite antihypertensive effect. Biochemical changes were seen in plasma renin activity, serum potassium and urate after the 12.5 and 25 mg dose. Three and 6 mg had no effect on blood pressure or metabolic parameters.

Left ventricular mass, serum electrolyte levels and cardiac arrhythmias in patients with mild hypertension treated with cilazapril or hydrochlorothiazide.

Thirty-five patients with mild hypertension (WHO Class I) participated in a double-blind cross-over study involving two 8-week periods of treatment with cilazapril 2.5-5 mg once daily or hydrochlorothiazide 25-50 mg once daily, in each case preceded by a 4-week placebo period. Thirty-two patients completed the study, the aim of which was to compare the effects of the drugs on serum electrolyte levels, left ventricular mass and cardiac arrhythmias, as assessed by echocardiography and 48-h Holter monitoring. Both drugs significantly reduced systolic (P < 0.01) and diastolic (P < 0.001) blood pressures (comparisons with placebo periods). Cilazapril and hydrochlorothiazide had opposite effects on ventricular ectopic activity. The beneficial effect of cilazapril on ventricular extrasystole counts correlated significantly (P < 0.001) with the reduction of left ventricular mass index. Hydrochlorothiazide had no effects on left ventricular mass or diastolic function. Serum potassium values were significantly (P < 0.001) reduced by hydrochlorothiazide but there was no correlation between changes in potassium levels and changes in ventricular ectopic activity. The results of the study suggest that hydrochlorothiazide and cilazapril were equally effective in reducing blood pressure, but only cilazapril reduced left ventricular hypertrophy and suppressed ventricular ectopic activity.

Sterility of male and female hybrids of Drosophila virilis and Drosophila lummei.

The sterility of interspecific hybrids between the sibling species Drosophila virilis and D. lummei was tested in reciprocal F1s and different second generation combinations of cytoplasm, sex chromosomes and autosomes. Males with motile sperm and females with at least one mature egg were scored as fertile. When D. virilis was the mother, about 5 per cent of the F1 male progeny was sterile, the reason being that the X of D. virilis was weakly incompatible with the heterozygous second, fourth and fifth chromosomes. The F1 males carrying the X chromosome of D. lummei were all fertile. The X chromosome of D. lummei, and in particular its species-specific double inversion In(1)a + b, nevertheless caused sterility in 70 per cent of males in the presence of homozygous autosomes of D. virilis. Sterility was rare among the females, although some weakly disadvantageous X versus autosome and autosome versus autosome interactions were detected. The male and female hybrid sterilities were based on different genetic systems. The results emphasize the central role of the X chromosome in hybrid male sterility.

Viability of first and second generation hybrids of Drosophila virilis and Drosophila lummei.

F1 hybrids of D. virilis and D. lummei survived significantly worse than the parents. When D. virilis was the mother, larger proportion of hybrids failed to emerge from pupal case, but overall survival rate of reciprocals was about the same. Viability of different heterospecific combinations of sex chromosomes and autosomes was studied in backcrosses. All the sex chromosomes survived well on the background of all-heterozygous autosomes. Hemizygous X was reciprocally subvital with homozygous alien autosomes. The reduced viability was male sex limited. The X-linked factor causing this effect was localized in the area of the large, phylogenetically ancient double inversion In(1)a+b. All four large autosomes were heterotic in backcross hybrids, more strongly in females. Five out of six possible first degree interactions between the autosomes were negative, indicating incompatibility between heterospecific autosomes. This effect was stronger in females. The genetic basis for hybrid subvitality is sex-specific.

Inheritance of male courtship sound characteristics in Drosophila littoralis.

Males of Drosophila littoralis vibrate their wings during courtship to deliver a "love song." This consists of 25- to 50-ms-long pulses with a basic frequency of about 250-400 Hz, separated by 250- to 500-ms pauses. When recording the sounds of flies from several localities in Europe, we found that males of one strain from northern Finland displayed courtship sounds with an unusually low wing beat frequency (below 250 Hz). In a genetic analysis utilizing marker stocks, the anomalous frequency was found to be caused by genes on all major autosomes, the strongest factors being on the second chromosome. Interaction between genes on chromosome 2 and on the fused chromosome 3-4 was non-additive. In low-frequency sounds, the number of cycles in the pulse (CN) was decreased, so that the length of the sound pulse (PL) remained more or less unchanged. We suggest that the genetically and physiologically most thoroughly controlled trait in the sound of Drosophila littoralis is the length of the pulse.

The effect of potassium and potassium plus magnesium supplementation on ventricular extrasystoles in mild hypertensives treated with hydrochlorothiazide.

Eleven mild hypertensives with base-line ventricular extrasystoles underwent a 24-week period of treatment with hydrochlorothiazide 50 mg daily. After 8 weeks of treatment either potassium as hydrochloride 1 g twice daily, or a combination of potassium hydrochloride 1 g and magnesium hydroxide 500 mg twice daily was added to the diuretic therapy in a randomised, double-blind, cross-over design. Each treatment period lasted 8 weeks. Supplementation with potassium or potassium plus magnesium resulted in a clear trend for suppression of ventricular ectopic activity. There was no difference in ventricular extrasystoles between the two supplementation groups. Supplementation with potassium plus magnesium but not with potassium only, resulted in a significant rise in serum potassium values (P less than 0.01). Our study suggests that potassium should be supplemented in thiazide-treated hypertensives who are prone to develop ventricular extrasystoles. Addition of magnesium to potassium supplementation does not result in further improvement in ventricular ectopic suppression.