Stress echocardiography in warmblood horses: comparison of dobutamine/atropine with treadmill exercise as cardiac stressors

The purpose of this study was to determine whether the combination of dobutamine and atropine causes cardiac stress equivalent to treadmill exercise. Therefore, electrocardiography and echocardiography were performed on 10 warmblood horses before, during, and after different cardiac stress tests. Stressors consisted of a standardized treadmill exercise and combined administration of dobutamine (7.5 microg/kg/min) and atropine (5 microg/kg). Maxima heart rates were achieved during the treadmill exercise (175 +/- 10 bpm). After exercise, a rapid decrease in heart rate was observed. Subsequently, a stress echocardiography for which a heart rate >100 bpm was required could only be performed within 1 minute after exercise. The mean heart rate during echocardiography was 136 +/- 8 bpm after exercise. The combination of dobutamine and atropine also resulted in a significant increase in heart rate, up to 141 +/- 20 bpm. Maxima heart rate was significantly higher during the treadmill exercise, but the decrease in heart rate was significantly slower after dobutamine and atropine administration. Over a period of 7.9 minutes, the mean heart rate was 123 +/- 8 bpm during dobutamine and atropine administration. Consequently, the combination of both drugs offered sufficient time for detailed examinations. Overall, echocardiographic examination identified a decrease in left ventricular (LV) dimensions, an increase in LV wall thickness, and a decrease in stroke volume after the treadmill exercise and during pharmacologic stress testing compared with baseline. Changes in echocardiographic variables generally were more pronounced during dobutamine and atropine administration. Similar to stress echocardiography in humans, in horses the combination of dobutamine and atropine is useful to produce an increase in heart rate comparable with what is achieved with exercise but without the need of increasing dobutamine dosage.     

Two-phase flavonoid formation in developing strawberry (Fragaria x ananassa) fruit

Flavonoids are important secondary metabolites in strawberry as they fulfill a wide variety of physiological functions. In addition, they are beneficial for human health. Previous studies have shown for selected enzymes from the flavonoid pathway that flavonoid biosynthesis shows two peaks during fruit development. We provide optimized protocols for the determination of the activities of the key flavonoid enzymes: phenylalanine ammonia lyase, chalcone synthase/chalcone isomerase, flavanone 3-hydroxylase, dihydroflavonol 4-reductase, flavonol synthase, flavonoid 3-O-glucosyltransferase, and flavonoid 7-O-glucosyltransferase. Using these protocols we were able to demonstrate two distinct activity peaks during fruit ripening at early and late developmental stages for all enzymes with the exception of flavonol synthase. The first activity peak corresponds to the formation of flavanols, while the second peak is clearly related to anthocyanin and flavonol accumulation. The results indicate that flavonoid 3-O-glucosyltransferase activity is not essential for redirection from flavanol to anthocyanin formation in strawberry.     

Preanalytic removal of human DNA eliminates false signals in general 16S rDNA PCR monitoring of bacterial pathogens in blood

PCR detection of microbial pathogens in blood from patients is a promising issue for rapid diagnosis of sepsis and early targeted therapy. However, for PCR assays detecting all bacterial groups, broad range primers, in particular the 16S rDNA targeting primers have to be used. Upcoming false signals and reduced sensitivity are a common problem as a consequence of unspecific amplification reactions with the human DNA background. Here we show that, using total DNA extracts from blood, unspecific signals occurred in general 16S rDNA PCRs as a result of the amplification of human sequences. To address this problem, we developed a protocol by which the human background DNA is removed and bacterial DNA is enriched during sample preparation, a method we termed background-free enrichment method (BFEM). In general, we aimed to exclude false signals due to the human background DNA yielded from 16S rDNA PCR, Real-Time-PCR and IGS-PCR analyses. We applied the BFEM to the analysis of blood samples from 22 patients and obtained results similar to standard blood culture methods. The BFEM allows specific and sensitive detection of pathogens in downstream PCR assays and is easy to handle due to the quick sample preparation procedure. Thus, the BFEM contributes to the generation of replicable and more reliable data in general 16S rDNA PCR assays.     

Exploring the natural variation for reproductive thermotolerance in wild tomato species

Climate change has become a serious threat for crop productivity worldwide. The increased frequency of heat waves strongly affects reproductive success and thus yield for many crop species, implying that breeding for thermotolerant cultivars is critical for food security. Insight into the genetic architecture of reproductive heat tolerance contributes to our fundamental understanding of the stress sensitivity of this process and at the same time may have applied value. In the case of tomato (Solanum lycopersicum), germplasm screenings for thermotolerance have often used yield as the main measured trait. However, due to the complex nature of yield and the relatively narrow genetic variation present in the cultivated germplasm screened, there has been limited progress in understanding the genetic basis of reproductive heat tolerance. Extending the screening to wild accessions of related species that cover a range of climatic conditions might be an effective approach to find novel, more tolerant genetic resources. The purpose of this study was to provide insight into the sensitivity of individual reproductive key traits (i.e. the number of pollen per flower, pollen viability and style protrusion) to heat-wave like long-term mild heat (LTMH), and determine the extent to which genetic variation exists for these traits among wild tomato species. We found that these traits were highly variable among the screened accessions. Although no overall thermotolerant species were identified, several S. pimpinellifolium individuals outperformed the best performing cultivar in terms of pollen viability under LTMH. Furthermore, we reveal that there has been local adaptation of reproductive heat tolerance, as accessions from lower elevations and higher annual temperature are more likely to show high pollen viability under LTMH.     

Female age as a factor determining the patterns of tree canopy utilisation by the European cherry fruit fly,Rhagoletis cerasi

The European cherry fruit fly, Rhagoletis cerasi is a univoltine, oligophagous pest of cultivated sweet cherries. The pest is closely associated with the canopy of its host tree, and its on-farm behaviour and mobility are determined by the crop structure and spatial arrangement of the farm. However, in spite of its relevance to integrated pest management, little is known about the within-canopy behaviour of R. cerasi, or the role of host and non-host canopy traits in local pest translocations. This paper reports the results of our exploratory studies, which reveal the modulating role of female age on various aspects of R. cerasi on-farm behaviour, such as diurnal and lifetime patterns of canopy utilisation and within-canopy activity, the propensity to undertake local explorative errands and the response to volatiles emanating from host and non-host canopies. The potential implications of our findings on the development of site-optimised IPM and/or pest-resilient agro-landscapes were also discussed.     

Allele-specific polymerase chain reaction diagnostic test for the functional MDR1 polymorphism in dogs

The major multidrug transporter P-glycoprotein (Pgp) contributes to the barrier function of several tissues and organs, including the brain. In a subpopulation of Collies and seven further dog breeds, a 4 base pair deletion has been described in the Pgp-encoding MDR1 gene. This deletion results in the absence of a functional form of Pgp and loss of its protective function. Severe intoxication with the Pgp substrate ivermectin has been attributed to the genetically determined lack of Pgp. An allele-specific polymerase chain reaction (PCR)-based screening method has been developed to detect the mutant allele and to determine if a dog is homozygous or heterozygous for the mutation. Based on this validation, the allele-specific PCR proved to be a robust, reproducible and specific tool, allowing rapid determination of the MDR1 genotype of dogs of at risk breeds using blood samples or buccal swabs.     

Soil development on loess substrate in central Germany — results of a long‐term trial on soil formation§

In 1948, a model soil formation trial was started in Halle/Saale by Schmalfuß. Loess nearly free of soil organic matter (SOM) was used as substrate and the soil formation has been studied in dependence on the level of mineral P and N fertilization. The trial consists of 48 glazed clay cylinders (∅︁ 40 cm × 100 cm), which are open at the bottom. The cylinders are embedded in the ground and stand on a layer of gravel. The trial is divided in a section with constant P and graduated N application and a section with constant N and graduated P application respectively. In the first 30 years, yields increased strongest in the highest NP fertilizer treatment compared with the no‐N or no‐P fertilizer treatment. Afterwards yields in the different fertilization treatments remained nearly constant on the reached levels. Actually the mean shoot dry matter per year is 2.5 times higher in the highest fertilizer treatment as compared to the no‐N fertilizer treatment. The C_org content in the topsoil increased over 49 years in the highest fertilized treatment from 0.18% at the beginning of the trial to 1.94%. Compared to the treatments without N or P fertilization this corresponded, however, only to a 1.2 fold increase. With higher levels of fertilizer application, humus accumulation and biological activity increased relatively faster than the C_org content in the topsoil. Soil organic matter improved the nutrient availability (increase of cation exchange capacity, mobile P fractions). It is assumed that decalcification and the decrease in the C/N ratio will continue. The lower C_org and N_org contents and the lower cation exchange capacity in comparison with soils developed in‐situ near to the parent loess material suggest a still ongoing soil formation process in the trial.     

Structure elucidation of omphalotin,a cyclic dodecapetide with potent nematicidal activity isolated from Omphalotus olearius

The structure and absolute configuration of omphalotin, a cyclic dodecapeptide isolated from the basidiomycete fungus Omphalotus olearius and possessing potent and selective activity against the plant pathogenic nematode Meloidogyne incognita, was determined by a combination of spectroscopic and chemical techniques. © 1998 Society of Chemical Industry     

Heat stress affects vegetative and reproductive performance and trait correlations in tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis>)

High ambient temperature has adverse effects on plant vegetative and reproductive development and reduces crop yield. To better understand the importance of male and female fertility for tomato fruit set ability under high temperature conditions and to test whether heat tolerance levels among and between reproductive and vegetative traits of genotypes correlate with each other, 13 tomato cultivars were subjected to long-term moderate heat (LTMH) or short-term heat shock (STHS), depending on the trait that was evaluated. LTMH caused significant decrease in performance of nearly all reproductive traits, i.e. pollen viability, pollen number, female fertility, seeded-fruit set and flower number per inflorescence, but not in inflorescence number. Considerable variation was found among cultivars, both under control and LTMH conditions. The cultivars Nagcarlang, Saladette and Malintka 101 produced a higher percentage of viable pollen under LTMH. For fruit set under LTMH condition, only cultivars that had been previously reported as being heat-tolerant produced fruits with seeds. STHS negatively affected vegetative traits concerning seedling survival and membrane stability. Correlation analysis revealed relationships between various traits within the control and heat treatments, but not between the two. Under heat stress fruit set was positively correlated with pollen viability, as well as with flower number per inflorescence. However, no significant correlations were found between vegetative and reproductive traits. Our data highlight the prominent role of pollen viability for tomato fertility under LTMH growth conditions. The observed variation in thermotolerance among different cultivars offers the possibility to decipher underlying physiological and genetic mechanisms.