Analysis of genetic diversity in the endangered tropical tree species <Emphasis Type="Italic">Hagenia abyssinica</Emphasis> using ISSR markers

Genetic diversity within and among 12 populations of the dioecious tropical tree species Hagenia abyssinica (Bruce) J.F. Gmel. in Ethiopia was examined with eight inter simple sequence repeat (ISSR) primers. A total of 104 clearly scorable bands were generated, among which 84 (81%) were polymorphic. Jaccard similarity coefficient was calculated for pairwise comparisons among all 120 individuals and ranged from 0.30 to 0.88 while average within-population similarity ranged from 0.53 to 0.66. Within-population variability was estimated as percentage polymorphic loci (ranging from 52% to 87%), Shannon’s information index (0.30–0.50) and Nei’s genetic diversity (0.21–0.35). The highest variability values were obtained for one recently planted population and for one wild population growing in an undisturbed primary forest area. Significant overall differentiation among populations was detected by both Shannon’s information index (0.26) and G _ST (0.25). Relatedness among samples was estimated with a principal coordinate analysis, and relatedness among populations was estimated with a cluster analysis (UPGMA). A Mantel test indicated a significant association between genetic and geographic distances, and an autocorrelation analysis showed significant evidence of gene flow over distances up to 30 km. This study is the first of its kind for H. abyssinica, which has decreased recently in Ethiopia and now must be regarded as an endangered species. Both within-population and between-population diversity estimates are typical of outcrossing, longlived and late successional species, suggesting that recent anthropogenic disturbances have not yet had much impact on population genetic parameters. DNA marker data can, however, be used to identify the most suitable sites for in situ conservation and for collection of material for establishment of genebanks and plant improvement programs.     

Integration of the rolA gene into the genome of the vigorous apple rootstock A2 reduced plant height and shortened internodes

Summary

The aim of this work was to dwarf the vigorous apple rootstock A2 by insertion of the rolA gene. To optimize conditions for a successful transformation, regeneration tests were carried out. The use of sucrose in the regeneration medium gave higher regeneration frequency than sorbitol in some cases and the shoot number per regenerated leaf was higher at 10 |j.M TDZ compared with 2.5 |j.M TDZ on the sucrose medium. Two transgenic clones, verified by PCR and Southern analysis, have been obtained on the sucrose medium together with 2.5 |j.M TDZ and 1.0 or 2.5 |j.M NAA and wounding by forceps. The two clones, named LAI and LA2, contained both the rolA and nptll genes. The results of in vitro rooting showed that LAI had a lower rooting percentage and a reduced root number per rooted shoot than the untransformed control shoots and the clone LA2 on the rooting medium containing 5 |JLM IBA. Growth analysis revealed that both transgenic clones had a reduced plant height and a shortened internode length compared with the control plants. However, the node number and the stem diameter were significantly larger for clone LAI than clone LA2 and the control plants.     

Pulsed delivery of inhaled nitric oxide counteracts hypoxaemia during 2.5 hours of inhalation anaesthesia in dorsally recumbent horses

Objective The study aimed to investigate the effect of varying pulse lengths of inhaled nitric oxide (iNO), and 2.5 hours of continuous pulse‐delivered iNO on pulmonary gas exchange in anaesthetized horses. Study Design Experimental study. Animals Six Standardbred horses. Methods Horses received acepromazine, detomidine, guaifenesin, thiopentone and isoflurane in oxygen, were positioned in dorsal recumbency and were breathing spontaneously. iNO was on average pulsed during the first 20, 30, 43 or 73% of the inspiration in 15 minute steps. The pulse length that corresponded to the highest (peak) partial pressure of arterial oxygen (PaO₂) in the individual horses was determined and delivered for a further 1.5 hours. Data measured or calculated included arterial and mixed venous partial pressures of O₂ and CO₂, heart rate, respiratory rate, expired minute ventilation, pulmonary and systemic arterial mean pressures, cardiac output and venous admixture. Data (mean ± SD) was analysed using anova with p < 0.05 considered significant. Results Although the pulse length of iNO that corresponded to peak PaO₂ varied between horses, administration of all pulse lengths of iNO increased PaO₂ compared to baseline. The shortest pulse lengths that resulted in the peak PaO₂ were 30 and 43% of the inspiration. Administration of iNO increased PaO₂ (12.6 ± 4.1 kPa [95 ± 31 mmHg] at baseline to a range of 23.0 ± 8.4 to 25.3 ± 9.0 kPa [173 to 190 mmHg]) and PaCO₂ (8.5 ± 1.2 kPa [64 ± 9 mmHg] to 9.8 ± 1.5 kPa [73 ± 11 mmHg]) and decreased venous admixture from 32 ± 6% to 25 ± 6%. The increase in PaO₂ and decrease in venous admixture was sustained for the entire 2.5 hours of iNO delivery. Conclusions The improvement in arterial oxygenation during pulsed delivery of iNO was significant and sustained throughout 2.5 hours of anaesthesia. Clinical relevance Pulsed iNO potentially could be used clinically to counteract hypoxemia in anaesthetized horses.     

Ventilation-perfusion relationships in the anaesthetised horse

Ventilation-perfusion relationships were studied by the multiple inert gas elimination technique in seven horses while they were conscious and during inhalation anaesthesia with halothane. A generally good match between ventilation and perfusion was found in the conscious, standing horse. During anaesthesia a huge shunt developed, ie perfusion of completely unventilated lung regions, both in dorsal and left lateral recumbency and whether the horse was breathing spontaneously or mechanically ventilated. The shunt was significantly greater and the arterial oxygen tension (PaO2) significantly lower in dorsal than in left lateral recumbency. Little or no perfusion of low VA/Q regions was observed during anaesthesia, whether ventilation was spontaneous or mechanical. Positive end-expiratory pressure (PEEP) did not significantly improve PaO2 or reduce the shunt. Selective mechanical ventilation of dependent lung regions with PEEP reduced the shunt markedly, an effect that was not achieved by conventional mechanical ventilation with general PEEP. The findings seem compatible with alveolar collapse during anaesthesia, causing shunt, whereas the absence of clearly low VA/Q regions questions the role of airway closure as the major disturbance of gas exchange.     

A REVIEW OF THE SONOGRAPHIC ASSESSMENT OF TUMOR METASTASES IN LIVER AND SUPERFICIAL LYMPH NODES

Diagnostic imaging techniques are an important part of the diagnostic workup and staging of cancer patients. Ultrasound is of particular interest in this respect. In so far as tumor metastases are concerned, ultrasonography of regional lymph nodes and of the liver can provide valuable information. In humans many criteria, some of them objective, have been evaluated as indicators of malignancy. The most diagnostically helpful of these include the short/long axis ratio of the lymph node, the pattern of distribution of the blood vessels within the lymph node, and to some extent the calculated values for resistive and pulsatility indices. Putative objective criteria to improve the specificity of ultrasound for metastases detection in the liver have also been evaluated. These include perfusion indices, primarily using analysis of Doppler frequencies (Doppler perfusion index) and hepatic venography using an ultrasound contrast agent. Contrast-enhanced ultrasonography is a new and promising area to help the initial diagnosis and characterization of malignancy, particularly for focal lesions in the liver. This review discusses the use of ultrasound for detection of metastases and presents material from four veterinary cases.     

Distribution and characterisation of fructan in wheat milling fractions

Structure and health effects of inulin-type fructans have been extensively studied, while less is known about the properties of the graminan-type fructans in wheat. Arabinoxylan (AX) is another important indigestible component in cereal grains, which may have beneficial health effects. In this study, the fructan content in milling fractions of two wheat cultivars was determined and related to ash, dietary fibre and AX contents. The molecular weight distribution of the fructans was analysed with HPAEC-PAD and MALDI-TOF MS using ¹H NMR and enzymatic hydrolysis for identification of fructans. The fructan content (g/100 g) ranged from 1.5 ± 0.2 in flour to 3.6 ± 0.5 in shorts and 3.7 ± 0.3 in bran. A correlation was found between fructan content and dietary fibre content (r = 0.93, P < 0.001), but with a smaller variation in fructan content between inner and outer parts of the grain. About 50% of the dietary fibre consisted of AX in all fractions. The fructans were found to have a DP of up to 19 with a similar molecular weight distribution in the different fractions.     

Evidence for seed transmission and symptomless growth of Ramularia collo‐cygni in barley (Hordeum vulgare)

Ramularia collo‐cygni (Rcc) is becoming an increasing problem for barley growers across Europe. However, the life cycle of the pathogen is only slowly being elucidated. In this study, Rcc DNA was detected in a number of harvested seed samples from 1999 to 2010, with mean levels peaking in winter barley samples in 2009. A number of experiments were carried out to determine whether the pathogen could move from barley seed to seedlings, and also from seed through the developing plant and into the subsequent generation of seed, both in controlled experiments and in field trials. Results from testing of seed indicated that the fungus is widespread at the end of the growing season in harvested grain samples and can be transmitted to developing plants from infected seed stock. Examination of infected seedlings did not reveal the presence of spores but fungal structures were found within the leaf. The location of the fungus within seed was examined, with Rcc DNA found in both embryo and non‐embryo tissue. The implications for barley production of the pathogen being seedborne are discussed.     

The effect of transdermal delivery of fentanyl on activity in growing pigs

Recently, decreased activity levels have been observed in pigs treated postoperatively with transdermal delivery of fentanyl (TD-fentanyl) after isoflurane anaesthesia. Whether the change in behaviour is related to opioid-induced sedation or to insufficient pain relief remains to be investigated. This study was therefore undertaken to evaluate the effect of TD-fentanyl 50 microg h(-1) on the activity level with and without isoflurane anaesthesia. Eight pigs (25.4 +/- 5.2 kg) were submitted to a cross-over study and given two treatments; 1) fentanyl patch applied after 30 minutes of anaesthesia (treatment A/F) and 2) fentanyl patch without anaesthesia (treatment F). The pigs' behaviour was observed from a video recording instantaneously every 10 minutes for 24 h before treatments and up to 72 h after the patch attachment. Venous blood samples were taken 1, 6, 12, 24, 48 and 72 h after the patch application. The behaviour recordings showed that TD-fentanyl did not produce sedation in any pig. No differences were found between the two treatments in activity level, weight gain or serum fentanyl concentration. This concentration measured after 24 h was 0.27 +/- 0.11 ng ml(-1) and 0.47 +/- 0.40 ng ml(-1) in the A/F and F group, respectively. In conclusion, transdermal delivery of 50 microg h(-1) fentanyl did not cause inactivity in growing pigs. However, the large variations in serum fentanyl concentration indicate that drug absorption from transdermal patches is unpredictable and sometimes deficient.