Characterisation of the first wheat (Triticum aestivum L.) nucleotide pyrophosphatase/phosphodiesterase resembling mammalian counterparts

A formerly unknown nucleotide pyrophosphatase/phosphodiesterase (NPP) of wheat (Triticum aestivum L.) (TaNPP) was recombinantly produced in Pichia pastoris (TaNPPr) following assembly of the sequence out of wheat ESTs using sequences of known NPPs. Simultaneously, a phosphodiesterase was purified to homogeneity from wheat germs, characterised and identified as TaNPP. TaNPP contains the highly conserved catalytic substrate and metal binding residues and displays properties [high pH optimum, glycosylation, high thermostability and inhibition by EDTA and adenosine 5′-monophosphate (AMP)] similar to those of mammalian enzymes characterised earlier. The sequence of TaNPP includes that of a putative transmembrane domain, a characteristic of most NPPs. Both recombinant and native TaNPP partially occur as oligomeric proteins on SDS PAGE. Upon addition of DTT, both migrate as monomeric proteins. Part of the native wheat protein even occurs in its truncated form, thus lacking the transmembrane region. Out of a range of tested natural substrates, TaNPP had the highest affinity towards adenine containing nucleotides. While Michaelis-Menten kinetics were valid in the low substrate concentration range, at higher concentrations, they were no longer applicable. TaNPP shows no similarity to the recently characterised rice and barley enzymes and, is hence, one of the first characterised plant NPPs resembling mammalian NPPs.     

An instructional model for training competence in solving clinical problems

We examined the design of a course that aims to ease the transition from pre-clinical learning into clinical work. This course is based on the premise that many of the difficulties with which students are confronted in this transition result from a lack of experience in applying knowledge in real practice situations. It is focused on the development of competence in solving clinical problems; uses an instructional model with alternating clinical practicals, demonstrations, and tutorials; and extends throughout the last pre-clinical year. We used a "proof-of-concept" approach to establish whether the core principles of the course design are feasible with regard to achieving the intended results. With the learning functions and processes as a frame of reference, retrospective analysis of the course's design features shows that this design matches the conditions from theories of the development of competence in solving clinical problems and instructional design. Three areas of uncertainty in the design are identified: the quality of the cases (information, openness), effective teaching (student and teacher roles), and adjustment to the development of competence (progress, coherence).     

Behavior of piglets after castration with or without carbon dioxide anesthesia

Surgical castration of male piglets without anesthesia is a routine management practice conducted on commercial pig farms. For animal welfare reasons, it would be beneficial to develop methods of practical pain relief. The objective of this study was to evaluate the effect of providing CO? anesthesia before castration on the behavior of piglets for up to 8 d after castration in comparison with piglets castrated without anesthesia. In 3 successive replicates, the behavior of 186 male piglets castrated with (n = 95) or without (n = 91) anesthesia was observed for up to 8 d after castration. All piglets in a given replicate were castrated on the same day, before 8 d of age. Behavioral observations were carried out in accordance with a continuous focal sampling procedure that began immediately after castration and continued for a period of 1 wk. Barrows anesthetized with CO? before castration displayed more interactive behaviors during the overall observation period than the other barrows (P = 0.0412), which may indicate better welfare. Assessing all observation periods separately, differences in activity at the udder, lying, walking, and interactive behaviors appeared to support the beneficial effect of providing CO? anesthesia before castration. However, these differences varied over time between treatment groups. The most important conclusion was that piglets castrated with or without CO? anesthesia displayed behaviors indicative of pain and discomfort for up to 6 d after castration. Therefore, additional analgesia may be necessary to eliminate the long-term pain caused by castration even in piglets anesthetized with CO? before castration.     

Differences between the ovine tonsils based on an immunohistochemical quantification of the lymphocyte subpopulations

In sheep, the pharyngeal first defence line against oral and inhaled antigens is organized in six tonsils. Since tonsils are regarded as secondary lymphoid tissue and part of the acquired immune system which is subjected to induction through contact with antigens, an evaluation of the different lymphocyte populations in tonsils is useful to determine a tendency of the specific tonsils to more inductive or more effective immunity. By means of immunohistochemistry, different lymphocyte populations were quantified and localized using a panel of eight antibodies, i.e. anti-CD45, anti-CD21, anti-CD2, anti-CD3, anti-CD4, anti-CD8, anti-WC1 and anti-Ki67. The CD21+ B lymphocytes were localized within the tonsillar lymphoid follicles. The CD2+/CD3+ T lymphocytes were numerous in the interfollicular regions and were aligned underneath and within the epithelium but were also observed at the CD21+ pole of the lymphoid follicles. Near the lingual and tubal tonsils, and the tonsil of the soft palate, the CD45+ cells around the seromucous glands and in the lamina propria were mainly CD3+ T cells. In all tonsils, the WC1+ gamma delta T cells formed a small lymphocyte population which harboured the lamina propria and the interfollicular region. The relative percentages of the different lymphocyte populations of the large palatine and pharyngeal tonsils, which are macroscopically the most developed, were comparable. In contrast, the lingual tonsil was significantly different from the other tonsils not only by its small size and lack of lymphoid follicles, but also by the lymphocyte populations. Based on the lymphocyte populations, the ovine tonsils can be divided in three groups with the tonsil of the soft palate, the tubal and paraepiglottic tonsil forming an intermediate between the palatine and pharyngeal tonsils as true tonsils on the one side, and the lingual tonsil as a scattered lymphocyte aggregation on the other side.     

Compost Amendment to Sandy Soil Affects Soil Properties and Greenhouse Tomato Productivity

Sandy soils, with low productivity, could be improved by compost application to sustain crop production. This study aimed to examine the effect of three compost types (vegetable, fruit and yard waste compost, garden waste compost, and spent mushroom compost) on basic properties of a loamy sand and greenhouse tomato productivity. Disturbed and intact soil samples were taken from a decade-long compost field experiment on loamy sand with three compost types at application rate of 30 m³ ha^?1 yr^?1 (7.5 ton ha^?1 yr^?1). The soils were characterized for chemical and physical properties. Tomato was planted in a greenhouse using soil samples from the field and vegetative and yield parameters (plant height, stem diameter, leaf number, and fruit yield), water productivity, and harvest index were evaluated. All compost types significantly increased soil total carbon, total nitrogen, pH, electrical conductivity and significantly decreased bulk density, with no effect on plant available water compared to the control. Fresh and dry fruit weights were significantly increased after compost addition. Plant height, leaf number, stem diameter, and total biomass did not significantly improve after compost addition. Spent mushroom compost had greater effect in improving tomato productivity. A decade-long application of composts on loamy sand improved basic chemical and physical properties which were reflected in increased fruit yield in tomato. Since no negative effect of compost was observed, we suggest that sandy soils may serve as a safe end use option for these composts and potentially support crop growth.     

Analysis of return flows in a tank cascade system in Sri Lanka

In Sri Lanka, irrigation reservoirs (tanks) are usually connected sequentially and form cascades along the landscape. A study was carried out in the Anuradhapura District in the dry zone of Sri Lanka to understand the role of return flows in such tank cascade systems. The water balance of a tank cascade system was estimated using hydrological data collected over a one-year period. The system was extended about 25 km along a river composed of three small reservoirs having the command area of 31, 55, and 55 ha, respectively. In this system, about 46% of seepage water from tanks entered the paddy fields of the command area. The crop consumed part of the water and the rest returned to the downstream tank through the drainage canals. Percolation loss in the command areas was low (3.6 mm/day) since a considerable portion of the percolation returned to the downstream tank. These results showed that return flows, which are generally disregarded in the water budget, contributed considerably to the water supply of the tank cascade system.     

Novel Alkaloids from Marine Actinobacteria: Discovery and Characterization

The marine environment is an excellent resource for natural products with therapeutic potential. Its microbial inhabitants, often associated with other marine organisms, are specialized in the synthesis of bioactive secondary metabolites. Similar to their terrestrial counterparts, marine Actinobacteria are a prevalent source of these natural products. Here, we discuss 77 newly discovered alkaloids produced by such marine Actinobacteria between 2017 and mid-2021, as well as the strategies employed in their elucidation. While 12 different classes of alkaloids were unraveled, indoles, diketopiperazines, glutarimides, indolizidines, and pyrroles were most dominant. Discoveries were mainly based on experimental approaches where microbial extracts were analyzed in relation to novel compounds. Although such experimental procedures have proven useful in the past, the methodologies need adaptations to limit the chance of compound rediscovery. On the other hand, genome mining provides a different angle for natural product discovery. While the technology is still relatively young compared to experimental screening, significant improvement has been made in recent years. Together with synthetic biology tools, both genome mining and extract screening provide excellent opportunities for continued drug discovery from marine Actinobacteria.     

Functional evaluation of the various calculation methods for precompression stress

Precompression stress (σ_p) is commonly used as an indicator for the load carrying capacity of a soil and its stress history. Several calculation methods have been proposed to determine σ_p, but they have proven to be not interchangeable. The aim of this study was to perform a functional evaluation of seven existing methods to test which method would be most recommended to assess soil strength in cultivated fields. This was achieved by investigating the relation of the obtained σ_p of undisturbed soil samples to other soil properties and stress history. Additionally, the robustness of the methods was evaluated by checking their sensitivity to perturbation of the measured strains. The two most commonly used methods, the Casagrande method and Gompertz model fit method, were found to be the least suitable to calculate σ_p. These methods lack robustness and do not yield a significant relation with the relevant soil properties or stress history. Better results were obtained with Intersecting linear regressions or at the inflection point of a fourth-degree polynomial.     

Capnocytophaga canimorsus

Capnocytophaga canimorsus is a commensal bacterium in the oral flora of dogs and cats. The bacterium is a zoonotic agent and has been isolated from humans, infected by dog or cat bites, scratches, licks or simply exposure to dogs or cats. Here the infectious agent, its pathogenicity and potential virulence factors, infection in animals and humans, diagnostic methods, prevalence, therapy and prevention are described. Suggestions for future research are given.     

Effects of soil mechanical treatments combined with bramble and bracken control on the restoration of degraded understory in an ancient beech forest

This paper describes the ground floor vegetation that developed four years after tillage implements in an ancient beech forest in central Belgium. The purpose of this study was twofold: (1) to explore whether tillage has a lasting effect on soil compaction and soil moisture as well as on vegetation characteristics; and (2) to analyse whether two distinct tillage treatments (rotary plough vs. disc plough), combined with vegetation control when necessary, have the same effect on soil compaction, soil moisture and plant establishment. Of the 29 species recorded, 15 showed a significant recovery after soil loosening in the studied forest area. Interestingly, different tillage treatments did not have the same influence on plant establishment. Treatment effects on soil structure and/or moisture status can be considered as causing the observed growth response.