Teaching small animal reproduction via virtual patients

Virtual patients have become an interesting alternative in medical education. Due to increasing demands regarding theoretical and clinical teaching and to improve an interdisciplinary approach, a new blended learning concept including virtual patients was developed and implemented in the veterinary curriculum of the Freie Universität Berlin. In the presented project, three virtual patients from the field of canine reproduction were developed. They focus on pregnancy diagnosis with suspected luteal insufficiency, pyometra and benign prostatic hyperplasia, respectively. The results of an evaluation by veterinary students of the 7th semester showed a high acceptance of virtual patients in a blended learning reproduction module in the interdisciplinary lectures. Students especially preferred videos, such as video lectures, hands-on videos and animations as well as a glossary for background information, to successfully and autonomously work on a virtual case. The content covered by the new modules that were developed in the context of this project is part of a spiral curriculum; they will be revised and enhanced during the clinical year.     

Limb‐sparing in dogs using patient‐specific,three‐dimensional‐printed endoprosthesis for distal radial osteosarcoma: A pilot study

Limb‐sparing for distal radial osteosarcoma has a high rate of complications. Using personalized three‐dimensional (3D)‐printed implants might improve outcome. The goals of this study were to optimize use of patient‐specific, 3D‐printed endoprostheses for limb‐sparing in dogs in the clinical environment and to report the outcome. This was a pilot study where five client‐owned dogs were enrolled. Computed tomography (CT) of the thoracic limbs was performed, which was used to create patient‐specific endoprostheses and cutting guides, and repeated on the day of surgery. Intra‐arterial (IA) carboplatin was introduced in the clinical management. Limb‐sparing was performed. Outcome measures were time required to produce the endoprosthesis and cutting guide, fit between cutting guide and endoprosthesis with host bones, gait analysis, size of the tumour, percent tumour necrosis, complications, disease‐free interval (DFI) and survival time (ST). Four dogs received IA carboplatin. Excessive tumour growth between planning CT and surgery did not occur in any dog. The interval between the CT and surgery ranged from 14 to 70 days. Fit between the cutting‐guide and endoprosthesis with the host bones was good to excellent. At least one complication occurred in all dogs. Two dogs were euthanized with STs of 192 and 531 days. The other dogs were alive with a follow up of 534 to 575 days. IA chemotherapy is a promising strategy to minimize the risk of excessive tumour growth while waiting for the endoprosthesis and cutting‐guide to be made. The design of the cutting‐guide was critical for best fit of the endoprosthesis with host bones.     

Diurnal patterns of water use in Eucalyptus victrix indicate pronounced desiccation-rehydration cycles despite unlimited water supply

Knowledge about nocturnal transpiration (E(night)) of trees is increasing and its impact on regional water and carbon balance has been recognized. Most of this knowledge has been generated in temperate or equatorial regions. Yet, little is known about E(night) and tree water use (Q) in semi-arid regions. We investigated the influence of atmospheric conditions on daytime (Q(day)) and nighttime water transport (Q(night)) of Eucalyptus victrix L.A.S. Johnson & K.D. Hill growing over shallow groundwater (not >1.5 m in depth) in semi-arid tropical Australia. We recorded Q(day) and Q(night) at different tree heights in conjunction with measurements of stomatal conductance (g(s)) and partitioned E(night) from refilling processes. Q of average-sized trees (200-400 mm diameter) was 1000-3000 l month(-1), but increased exponentially with diameter such that large trees (>500 mm diameter) used up to 8000 l month(-1). Q was remarkably stable across seasons. Water flux densities (J(s)) varied significantly at different tree heights during day and night. We show that g(s) remained significantly different from zero and E(night) was always greater than zero due to vapor pressure deficits (D) that remained >1.5 kPa at night throughout the year. Q(night) reached a maximum of 50% of Q(day) and was >0.03 mm h(-1) averaged across seasons. Refilling began during afternoon hours and continued well into the night. Q(night) eventually stabilized and closely tracked D(night). Coupling of Q(night) and D(night) was particularly strong during the wet season (R2?=?0.95). We suggest that these trees have developed the capacity to withstand a pronounced desiccation-rehydration cycle in a semi-arid environment. Such a cycle has important implications for local and regional hydrological budgets of semi-arid landscapes, as large nighttime water fluxes must be included in any accounting.     

Rapid monitoring of species abundance for biodiversity conservation: Consistency and reliability of the MacKinnon lists technique

Effective monitoring of biodiversity for conservation requires information on spatial and temporal variation in species’ abundances. As conservation resources are limited, monitoring methods are required that enable rapid and cost effective data collection. There are many traditional methods of estimating absolute abundance, such as territory mapping and distance sampling. However, these typically require more time, expertise and finances than are available across much of the globe. This is especially so in the tropics, where high species richness, low densities of many species and structurally complex environments also make monitoring particularly challenging. The MacKinnon lists technique is a rapid assessment methodology designed for use in species rich environments. This method is typically used to estimate species richness, but it has also been suggested that it can generate consistent abundance indices, even when observer experience and environmental conditions vary. If this suggestion is correct, the MacKinnon lists method could be used to assess spatial or temporal changes in abundance using diverse survey data. Here, we provide the first detailed assessment of intra- and inter-observer consistency of the Mackinnon List method in generating species abundance indices that could be useful for conservation monitoring purposes. As a case study, we use one of the world’s most diverse avifaunas, that of the forested Bolivian Andes. We show that MacKinnon lists can provide species abundance indices that are consistent between observers of markedly different experience of the focal avifauna (zero to six years), and between assessments carried out in different stages of the breeding season, between which detectability of individuals differed significantly. We believe this is the first time that a biodiversity monitoring method has been demonstrated to produce consistent abundance indices for a highly diverse avian tropical assemblage. We also suggest that the MacKinnon lists methodology has the potential to be a very useful conservation monitoring tool for many taxa in species rich environments, such as the tropics.     

Determination of Chemical Composition and Nutritive Value with Fatty Acid Compositions of African Mangosteen (<Emphasis Type="Italic">Garcinia Livingstonei</Emphasis>)

African mangosteen (Garcinia livingstonei T. Anderson) is native to Africa and is exotic to other parts of the world. It is a fruit bearing tree with multiple uses. This study was purposed to determine, by analysis, the chemical compositions and nutritive value of the fruits of African mangosteen. Proximate compositions varied with the portions such as epicarp, mesocarp, endocarp and seed tested. All fruit portions contained carbohydrate (37.67–95.02%) and crude protein (0.65–31.76%) as their major components. Moisture (0.45–3.42%), crude fat (1.23–19.55%), crude fiber (2.93–21.13%) and ash (1.76–5.44%) were also found at different levels, depending upon the portions. All portions of fruit were rich in macro and micro elements. The fruits also possessed phenolics (174.02–10.725?mg GAE per g), flavonoid (19.25 to 99.98?µg QE per g) and alkaloid (1.56 to 9.49?mg/kg) contents. The constitution of tannins and oxalate which contribute towards the anti-nutritive value was also low, thus making the fruits beneficial for consumption. Quality attributes like relative amount of oleic acid, total unsaturated fatty acid, acid value, free fatty acid, peroxide value and iodine value showed that the seed oil of African mangosteen can become one among the oleic acid based vegetable oils for consumption.     

A multi-metric index for assessing two decades of community responses to broad-scale shoreline enhancement and restoration along the Toronto waterfront

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Effect of wide spacing on tree growth, branch and sapwood properties of young Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] in south-western Germany

The influence of stand density on Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] is conceptually understood, but for wide spacings not well quantified, particularly in Europe. This study used 41 trees from 7 different locations in south-western Germany to compare important tree- and branch-level attributes across three different densities, namely 100, 200, and 1,200 stems ha⁻¹. In general, there were only a few tree and branch attributes that were significantly different between the 100 and 200 ha⁻¹ densities. Crown projection area and diameter of the thickest branches were the most important differences between the 100 and 200 ha⁻¹ densities. The most obvious and significant differences in this study were between 100 and 1,200 ha⁻¹ densities, where nearly every examined tree and branch attributes were statistically significant. However, relative sapwood area, the number of branches, branch angle, and the occurrence of spike knots were insensitive to stand density. Although the two lowest stand densities in this study represent rather extremely wide spacings, these results still have important implications for the development of effective thinning regimes for Douglas-fir in south-western Germany. Important management recommendations from this study include thinned stands should be maintained to at least 200 stems ha⁻¹ to maintain high log quality and stand stability. Furthermore, even at stand densities exceeding more than 1,200 trees ha⁻¹ planted trees, artificial pruning may even be necessary to produce high quality logs.     

Discrimination of alleles and copy numbers at the <Emphasis Type="Italic">Q</Emphasis> locus in hexaploid wheat using quantitative pyrosequencing

Speltoid spikes are characterized by pyramidal spike morphology featuring an elongated rachis and tenacious glumes. Speltoids are considered undesirable spike aberrants in wheat breeding leading to increased heterogeneity within a cultivar candidate. As a consequence, the presence of speltoids may result in rejection of a cultivar candidate during official field trials or denial of cultivar certification during seed multiplication. A reliable method is, thus, required to assess the occurrence of speltoids, early on in a wheat breeding program. The domestication gene Q located on the long arm of wheat chromosome 5A is known to suppress the speltoid phenotype in wheat. Here, a quantitative pyrosequencing assay was developed to distinguish between normal wheat plants, which possess two copies of the Q allele, and aberrants, which are either aneuploids lacking the correct number of chromosome 5A copies or plants which carry the primitive q allele. An accurate and reproducible determination of the Q gene copy number was achieved for different wheat genotypes based on homoeologous sequence quantification with two primer combinations at the Q locus. Single plants with one to four copies of the Q allele could be detected by quantitative pyrosequencing which corresponded to the occurrence of speltoid (1 Q allele), normal (2 Q alleles), and compact (more than 2 Q alleles) spikes. Q and q specific alleles could be differentiated at SNP position 2299 of the Q gene. This SNP is assumed to be related to the emergence of free-threshing wheat forms. To our knowledge this is the first report for detection of aneuploids and differentiation of Q alleles in bread wheat using pyrosequencing technology. In future, quantitative pyrosequencing assay can be applied in wheat breeding programs to carry out marker-assisted selection against the presence of speltoid spike aberrants.     

Production of complex human glycoproteins in yeast

We report the humanization of the glycosylation pathway in the yeast Pichia pastoris to secrete a human glycoprotein with uniform complex N-glycosylation. The process involved eliminating endogenous yeast glycosylation pathways, while properly localizing five active eukaryotic proteins, including mannosidases I and II, N-acetylglucosaminyl transferases I and II, and uridine 5'-diphosphate (UDP)-N-acetylglucosamine transporter. Targeted localization of the enzymes enabled the generation of a synthetic in vivo glycosylation pathway, which produced the complex human N-glycan N-acetylglucosamine2-mannose3-N-acetylglucosamine2 (GlcNAc2Man3GlcNAc2). The ability to generate human glycoproteins with homogeneous N-glycan structures in a fungal host is a step toward producing therapeutic glycoproteins and could become a tool for elucidating the structure-function relation of glycoproteins.