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Two major mtDNA lineages of brown trout come into contact in the upper Danube basin of Central Europe. The region is additionally confronted with stock management programs that often use domesticated foreign strains creating confusion as to what constitutes native and non-native populations as well as how to define major management units. In a unique system in north eastern Austria we provide additional support that the so-called Atlantic lineage of brown trout is native to the upper Danube and provide a high resolution microsatellite screening protocol that clearly distinguishes between natural and anthropogenic introgression, with the latter being minimal. Sequencing of 6,000 bp of mtDNA also shows that domesticated stocks primarily stem from a distinct albeit genetically diverse sub-clade of the Atlantic lineage, approximately 50,000-100,000 years divergent from the native Atlantic stocks, thought to have expanded into the region during the Holocene. The futility in defining large-scale management units based on mtDNA lineages for such broadly distributed and highly fragmented species is discussed. [ABSTRACT FROM AUTHOR]

Fruit quality traits are major breeding targets in the Rosaceae. Several of the major Rosaceae species are current or ancient polyploids. To dissect the inheritance of fruit quality traits in polyploid fleshy fruit species, we used a cultivated strawberry segregating population comprising a 213 full-sibling F1 progeny from a cross between the variety 'Capitola' and the genotype 'CF1116'. We previously developed the most comprehensive strawberry linkage map, which displays seven homoeology groups (HG), including each four homoeology linkage groups (Genetics 179:2045-2060, ). The map was used to identify quantitative trait loci (QTL) for 19 fruit traits related to fruit development, texture, colour, anthocyanin, sugar and organic acid contents. Analyses were carried out over two or three successive years on field-grown plants. QTL were detected for all the analysed traits. Because strawberry is an octopolyploid species, QTL controlling a given trait and located at orthologous positions on different homoeologous linkage groups within one HG are considered as homoeo-QTL. We found that, for various traits, about one-fourth of QTL were putative homoeo-QTL and were localised on two linkage groups. Several homoeo-QTL could be detected the same year, suggesting that several copies of the gene underlying the QTL are functional. The detection of some other homoeo-QTL was year-dependent. Therefore, changes in allelic expression could take place in response to environmental changes. We believe that, in strawberry as in other polyploid fruit species, the mechanisms unravelled in the present study may play a crucial role in the variations of fruit quality. [ABSTRACT FROM AUTHOR]

Purpose: Investigating the diagnostic value of color Doppler ultrasound for defining the varicocele grade according to WHO criteria. Methods: A total of 217 men (129 with clinical varicocele and 88 without clinical varicocele) were investigated by physical examination and color Doppler ultrasound and categorized according to WHO varicocele criteria (0, subclinical, I, II, and III). Diameter and reflux of the largest vein in the pampiniform plexus were measured bilaterally with the patient in the supine position in rest and during the Valsalva maneuver. To assess the possibility of differentiating varicocele grade by venous diameter, optimal cut-point values were determined by receiver-operator characteristic (ROC) analysis. Results: With increased varicocele grade, a larger vein diameter was more significant in rest and during Valsalva (in all cases P < 0.05), except between grade I and grade II. Retrograde peak flow velocities were similar in every group (in all cases P > 0.1). Only grade III varicoceles demonstrated significantly increased peak flow values compared with all other grades ( P < 0.001). There were no side-related differences when comparing identical varicocele grades (in all cases P > 0.1). Venous diameters above 2.45 mm in rest (sensitivity 84%, specificity 81%) or 2.95 mm during Valsalva (sensitivity 84%, specificity 84%) predicted the presence of a clinical varicocele. Conclusions: Our findings support the hypothesis that clinical varicoceles can be predicted with high accuracy based only on the diameter of testicular veins using cut-point values of >2.45 mm in rest or >2.95 mm during Valsalva maneuver in the supine position. [ABSTRACT FROM AUTHOR]

Bulked segregant analysis combined with AFLPs was used to identify molecular markers linked to the Rca2 gene conferring resistance to Colletotrichum acutatum pathogenicity group 2 which causes anthracnose in the octoploid strawberry Fragaria × ananassa. DNA bulks originating from a cross between the resistant cultivar ‘Capitola’ and the susceptible cultivar ‘Pajaro’ were screened with 110 EcoRI/ MseI AFLP combinations. Four AFLP markers were found linked in coupling phase to Rca2 with recombination percentages between 0% and 17.7%. Among the four markers linked to the resistance gene, two were converted into SCAR markers (STS-Rca2_417 and STS-Rca2_240) and screened in a large segregating population including 179 genotypes. The Rca2 resistance gene was estimated to be 0.6 cM from STS-Rca2_417 and 2.8 cM from STS-Rca2_240. The presence/absence of the two SCAR markers was further studied in 43 cultivars of F. × ananassa, including 14 susceptible, 28 resistant, and one intermediate genotype. Results showed that 81.4% and 62.8% of the resistant/susceptible genotypes were correctly predicted by using STS-Rca2_417 and STS-Rca2_240, respectively. The 14 susceptible genotypes showed no amplification for either SCARs. These developed SCARs constitute new tools for indirect selection criteria of anthracnose resistance genotypes in strawberry breeding programs. [ABSTRACT FROM AUTHOR]

A two-way pseudo-testcross strategy, combined with Single Dose Restriction Fragment (SDRF) marker analysis, was used for genetic mapping in the octoploid cultivated strawberry Fragaria × ananassa (2n = 8x = 56). Based on a 113 full-sib progeny from a cross between the variety Capitola and the clone CF1116, we generated two parental maps using Amplified Fragment Length Polymorphism (AFLP) markers. Ninety two percent of the markers (727 out of 789) showed ratios corresponding to simplex markers (the majority being SDRF markers), and 8% (62 out of 789) fitted a multiplex ratio. Linkage maps were first established using SDRF markers in coupling phase. The female map comprised 235 markers distributed among 43 co-segregation groups, giving a map size of 1,604 cM. On the male map, 280 markers were assigned to 43 co-segregation groups, yielding a map size of 1,496 cM. Once the co-segregation groups were established, their association was tested using repulsion-phase markers. In total, taking into account associations representing the same linkage groups, 30 linkage groups were detected on the female side and 28 on the male side. On the female map, 68.3% of the pairwise marker linkages were in coupling versus 31.7% in repulsion phase, and the corresponding figures on the male map were 72.2% and 27.8%, respectively. In addition, both groups linked only in the coupling phase and groups linked in the repulsion phase were characterized. The observations suggest that the meiotic behavior of the F. × ananassa genome is neither fully disomic nor fully polysomic, but rather mixed. The genome may not be as completely diploidized as previously assumed. [ABSTRACT FROM AUTHOR]

We have constructed nearly complete linkage maps of Pinus svlvestris (L.) using AFLP markers based on a two-way pseudo-testcross strategy in a full-sib family founded in an advanced breeding program. With 39 primer combinations, a total of 737 markers (320 from the mother and 417 from the father) segregated in a 1:1 ratio, corresponding to DNA polymorphism: heterozygous in one parent and null in the other. In the maternal parent, 188 framework markers were mapped in 12 linkage groups, equivalent to the Pinus haploid chromosome number, with a total coverage of 1,695.5 cM. In the paternal parent, 245 framework markers established a map with 15 linkage groups, spanning a genome length of 1,718.5 cM. The estimated total map length was LF = 1,681 cM for the female and LM = 1,645 cM for the male using a modified method-of-moment estimator. Combining these values with those estimated from the observed map lengths in both parents, we estimated the genome length in Scots pine to be between 1 ,600 and 2,100 cM. Our genome coverage was estimated to be more than 98% with a framework marker interval of 20 cM for both parents. Most of the female and male linkage groups were associated through the analysis of the intercross markers. [ABSTRACT FROM AUTHOR]

The aim of the present study was to evaluate the diagnostic accuracy of serology by using new assays for the detection of genus and species-specific lgG, IgM, IgA and secretory IgA antibody in female sex workers. Cervical swabs and first void urine (FVU) from 314 female sex workers were submitted to nucleic add amplification by ligase chain reaction (LCx, Abbott). Concomittantly, blood samples were tested for the presence of lgG, IgM and IgA antibodies using a genus-specific assay (rELISA, Medac) and species-specific test (SeroCT, Orgenics). Chlamydia trachomatis infection was detected in a total of 30 (9.6%) female sex workers by LCR. With rELISA, seroprevalences for lgG, IgM, and IgA antibody to Chlamydia were 88.9%, 19.1% and 62.7%, respectively. lgG and IgA antibody prevalences against C. trachomatis (SeroCT) were 65.0% and 23.9%, respectively. In comparison to the positive LCR results obtained from cervical swab and/or FVU, the sensitivity of rELISA for Chlamydia lgG, IgA and IgM detection was 93.3%, 83.3% and 16.7%, respectively. With SeroCT, the sensitivity for C. trachomatis-specific lgG and IgA detection was 86.7% and 33.3%, respectively. The specificities of both serologic tests in comparison to LCR were very low. In conclusion, the correlation of serology with active C. trachomatis infection of the lower genital tract is very low. According to our results, serologic testing for Chlamydia can exclude active infection of the lower genital tract with a high reliability (≥ 95%). However, detection of C. trachomatis can only be reliably achieved by nucleic acid amplification assays. [ABSTRACT FROM AUTHOR]

The nicotinamide antagonist 6-aminonicotinamide (6-AN) administered to rats prior to the second postnatal week drastically affected their motoric activity by producing consecutive reversible hypo- and hypermotoric stages. To obtain information on the validity of the putative link of correlated changes in brain RNA synthesis and motoric activity, neuronal and nonastrocytic glial cerebral and cerebellar in vitro RNA synthesis was determined during the hypo- and hypermotoric stages induced by in vivo administration of 6-AN given on the 7th day of a rat's life. Significant changes were apparent in the rate of neuronal cerebral and cerebellar RNA synthesis coinciding with the different phases of motor activity. However, only the changes in neuronal cerebral RNA synthesis were directly proportional to the changing pattern of motor activity. Measurement of the total number of RNA initiation sites present in neuronal cerebral chromatin, which was isolated during the phase of reduced and elevated muscular activity, revealed that the drug-induced changes in neuronal cerebral RNA synthesis could be directly attributed to a genetically repressed or activated state of the corresponding neuronal chromatin fraction. Nevertheless, although there was evidence suggesting a connection between changes in neuronal cerebral RNA synthesis and corresponding changes in muscular activity it was difficult to deduce any direct relationship between parameters owing to the absence of a convincing biological assay for assessing the role of RNA in motor function. [ABSTRACT FROM AUTHOR]

To determine the local gastrointestinal absorption of a new synthetic somatostatin analogue (SMS 201-995 = Sandostatin), an intestinal tube was passed in eight healthy volunteers and on different days an aqueous solution was administered at four different locations: stomach, proximal duodenum, ligament of Treitz and jejunum. In a follow-up study, an oro-ileal tube was passed in six of the original volunteers and the drug solution was administered in to the terminal ileum. The aqueous solution of SMS was rapidly absorbed from the gastrointestinal tract after local application, and it was well tolerated. Absorption of the drug from the different sites was comparable, although there was a tendency to decreased peptide absorption after ileal administration. Absorption of the drug was quite variable between the subjects and the different locations. The dose-corrected systemic availability relative to subcutaneous administration in another study was 0.28%. However, significant plasma SMS concentrations were achieved, suggesting that oral delivery of the polypeptide may eventually be possible for long-term treatment of a variety of disorders. [ABSTRACT FROM AUTHOR]

Screening for apolipoprotein (apo) C-II variants in the plasma of 400 students, 600 patients of a cardiological rehabilitation center, and 1200 patients of an outpatient lipid clinic by isoelectric focusing and subsequent anti-apo C-II immunoblotting led to the identification of four individuals whose plasma samples contained an apo C-II isoform with an abnormal isoelectric point. In all cases direct sequencing of PCR-amplified DNA assessed a heterozygous A to C transversion in codon 19 of the apo C-II gene which leads to the replacement of lysine with threonine. Two of the four index patients presented with moderate hypertriglyceridemia; one suffered from severe hyperlipidemia, with triglyceride levels ranging between 180 and 1900 mg/dl, depending on dietary changes. Sequencing of this proband's lipoprotein lipase gene showed no alteration compared to the wildtype sequence. A study in his family revealed that heterozygosity for apo C-II(K19T) is not associated with differences in mean lipid and lipoprotein concentrations. In conclusion, apo C-II(K19T) occurs in Germany at a frequency of approximately 1 in 550. Although this variant is not sufficient to cause hypertriglyceridemia, it may be possible that apo C-II(K19T) causes hypertriglyceridemia in the presence of additional as yet unidentified environmental and/or genetic factors. [ABSTRACT FROM AUTHOR]

Two monoclonal Beta-cell surface antibodies M10H6 und K14D10 were obtained by fusion of spleen cells of Balb/c mice with the myeloma cell line P0. The monoclonal antibody M10H6 was induced by immunization with rat insulinoma cells finally boostered with disintegrated rat islets, whereas the K14D10 was generated after immunization with porcine proinsulin. Both monoclonals belong to the IgG isotype and were screened with insulin-producing rat insulinoma cells by an indirect immunofluorescence test as well as by a cellular enzyme linked immunosorbent assay. In addition to the cell surface binding on living Beta cells the monoclonals react with islets on cryostat sections of rat pancreas. The anti-islet cytotoxic potential of these monoclonals was measured by Chromium-release in the presence of complement or Fc-receptor bearing leucocytes using Chromium-labelled rat islet cells as target. Both antibody secreting hybridomas were propagated in syngeneic mice resulting in high levels of islet cell surface antibodies in ascites and sera from the recipient. High anti-islet cytotoxicity was mediated by ascites fluid, but no mouse developed hyperglycaemia. Furthermore, the repeated injections of the monoclonals into rats did not exert a diabetogenic action and failed to reduce the pancreatic insulin content although the attraction of the K14D10 to the pancreatic islets in vivo could be demonstrated. We conclude that islet cell surface antibody-mediated Beta-cell lysis in vitro may not be relevant to Beta-cell destruction in vivo. [ABSTRACT FROM AUTHOR]

Isolated pancreatic islets of normoglycemic sand rats do not respond to 2.5 mM glucose with an enhanced glucagon secretion, which could be observed in normal Wistar rats. Arginine stimulates glucagon release in the presence of 2.5 mM glucose in Wistar rats as well as in sand rats. The secretion pattern is not caused by insulin deficiency since sand rat islets are characterized by an increased insulin secretion rate in vitro. This paradoxical glucagon secretion is not caused by a changed glucagon content but might be related to this species which is able to develop a diabetic syndrome spontaneously. [ABSTRACT FROM AUTHOR]

Muonic Coulomb capture ratios of NaO, CaO, RbO, SrO, CsO and BaO have been measured and compared with theoretical predictions. Muonic x-ray intensities have been used to determine the angular momentum distribution of the muons for the principal quantum number n=20. A systematic variation of this distribution with Z was observed. [ABSTRACT FROM AUTHOR]

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On day 13+0 h of gestation rats received 10 mg/kg cyclophosphamide i.p. 3, 9, and 24 h after administration of the drug the activities of the DNA-dependent RNA polymerase reactions were measured in the whole embryo and in parts of it (head, upper and lower extremities and trunk). The activities of these enzymes were correlated with the increase in the RNA content of these tissues. A relationship was established light and electron microscopically between the biochemical results and the specific alterations which occur in different structures of the embryonic tissue. The results of these investigations show that: In this paper a disturbance of the transcription mechanisms is discussed as being the primary mode of action of cyclophosphamide in cell metabolism. [ABSTRACT FROM AUTHOR]

In order to elucidate the phase specificity of the embryo-lethal and teratogenic action of actinomycin D (day 5-10 of gestation) rats received H-actinomycin D at different times of gestation. The distribution of the radioactivity in the fetus and the placenta was measured. Moreover, the effects of actinomycin D on the RNA polymerase reactions in isolated nuclei of the decidual and fetal tissue were studied. The biochemical results were correlated electron microscopically with specific alterations occurring in different structures of the total implantation sites. These studies show that the time-dependent embryo-lethal and teratogenic effects of actinomycin D result from a lesion of the embryo and the decidua. The effects of the decidua occur at an earlier time and are more pronounced as compared to those of the embryo. Since the decidua is significant for the nutrition of the embryo till closure of the yolk-sac and beginning of the yolk-sac circulation (day 10 of gestation), the direct effects of actinomycin D on the embryo are intensified by the lesion of the decidua. After day 10 of gestation till the beginning of the placental function H-actinomycin D passes over to the embryo. But the concentration-even after a dose of 1 mg/kg-does not suffice to inhibit in vivo RNA polymerase reactions in isolated nuclei. [ABSTRACT FROM AUTHOR]

The increase in DNA, RNA, and nitrogen as well as the activity of some enzymes involved in nucleic acid metabolism were measured during liver development. During the perinatal period (day 18 of gestation until birth) high activities of thymidine kinase and DNA polymerase were found which decline in the early suckling period, whereas the activities of the DNA-dependent RNA polymerases show a slow continuous increase. During the juvenile period (starting about 4 weeks after birth) the values of all enzymes studied approximate to those measured in liver tissue of adult animals. The correlation of these enzymes with the growth kinetics of liver tissue is discussed. [ABSTRACT FROM AUTHOR]

Isolated, electrically stimulated left guinea-pig atria were incubated at 30° C in a Tyrode solution, the hydrogen ion concentration of which was varied by altering the percentage of the aerating CO/O-mixture. The increase of the extracellular hydrogen ion concentration from pH 8.0 to pH 7.0 caused a reduction of the calcium uptake and of the contraction force. The positive inotropic effect and the concomitantly occuring increased calcium uptake induced by isoprenaline and tyramine were markedly diminished at the lower pH, the response to tyramine being more affected by increased hydrogen ion concentration than that of isoprenaline. This is possibly due to a decrease in noradrenaline available at pH 7.0 for release by tyramine. Isoprenaline and tyramine enhanced the relative specific calcium activity (RSA) without altering the total tissue calcium content. On the contrary, the increase of the extracellular calcium concentration caused an augmentation of the relative specific calcium activity and the total tissue calcium content. [ABSTRACT FROM AUTHOR]

In order to get insight into the synthesis rates occuring in mammalian embryos during the phase of differentiation, wet and dry weight as well as DNA-, RNA-, and nitrogen content of the whole implantation site (day 7 to 10 of gestation) and of the embryos (day 11 to 14) have been determined. Growth of the whole implantation site (day 7 to 10) proceeds almost linearly: Within a 24-h period the weight increases about 2-fold. The growth of the embryos, however, is found to be different in each 24-h period between day 8 and 14. All synthesis processes proceed at an extremely rapid rate between day 9 and 12 of gestation. The weight of embryos shows a 6-fold rise between day 11 and 12, the increase being even more pronounced on the preceding days. Growth and increase in cell mass are essentially due to cell division, since DNA content and weight change in a parallel way. Data on the nitrogen content per embryo show that the protein synthesis keeps pace with the rapid cell division. Using pure nuclear fractions isolated from 11- to 14-day-old embryos the average distribution of RNA within the cells can be estimated. The increase in the number of cells within a 24-h period is calculated from the average DNA content per embryo and the average DNA content per nucleus measured in isolated nuclei. The DNA content per nucleus of 12- to 14-day-old rat embryos is found to be almost twice (12±0.5 pg) that of diploid nuclei (6.7-7.2 pg). This inducates that a high percentage of the cells are in the late S-phase or in the G-phase. The growth rate of the embryos between day 8 and 14 of gestation is calculated from the 24-h rate of increase in cell number which is obtained either biochemically or in the earlier stage of embryonic development with morphological studies. Some peculiarities of the duration of the cell cycle of the rapidly growing tissue are discussed. [ABSTRACT FROM AUTHOR]

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The effects of estrone plus progesterone on embryonic survival were studied in hypophysectomised pregnant rats. Replacement therapy commenced 24 h prior to the ablation of the pituitary which was performed on day 6 of pregnancy. The number of viable fetuses was correlated with increasing amounts of progesterone from 24 to 84 mg/kg given in concert with a constant dose of 4 μg/kg estrone. As judged by the DNA content of the 14-day-old embryos, growth retardation was linked to the reduction of the progesterone levels. No visible malformations were recorded although the experimental period covered the phase of organogenesis. On the other hand, when measuring in isolated nuclei of 14-day-old rat embryos the activity of DNA polymerase, an enzyme the activity of which can be correlated to a certain extent to embryonic growth, it becomes obvious that after treatment with estrone in combination with low (24 or 36 mg/kg) or too high doses (84 mg/kg) of progesterone the correlation between DNA content of the embryos and DNA polymerase activity per cell diverges. This result may point to a cellular malfunction not disclosed by the macroscopic appearance of the embryos at this stage. So it can be concluded that embryonic growth and differentiation processes might be directly dependent on maternal gonadal hormones, at least till day 12 of pregnancy. [ABSTRACT FROM AUTHOR]

In closed-chest cats, anaesthetized with pentobarbital, a comparative study was carried out of the cardiovascular actions of four β-receptor blocking agents [propanolol, practolol, d(-) INPEA and LB 46] as well as of their antagonism to the effects of isoproterenol. The β-receptor blocking agents were injected intravenously in increasing doses until the positive inotropic action of isoproterenol was abolished. Changes in maximum rate of rise of ventricular pressure ( dp/dt max) were taken as a measure of the inotropic action. Heart rate, enddiastolic ventricular pressure, ventricular pressure, arterial pressure and ECG were recorded. The following results were obtained: [ABSTRACT FROM AUTHOR]

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