Influence of Fucoidan Extracts from Different Fucus Species on Adult Stem Cells and Molecular Mediators in In Vitro Models for Bone Formation and Vascularization

Fucoidans, sulfated polysaccharides extracted from brown algae, are marine products with the potential to modulate bone formation and vascularization processes. The bioactivity and safety of fucoidans are highly associated with their chemical structure, which may vary with algae species and extraction method. Thus, in depth evaluation of fucoidan extracts in terms of endotoxin content, cytotoxicity, and their detailed molecular biological impact on the individual cell types in bone is needed. In this study, we characterized fucoidan extracts from three different Fucus species including Fucus vesiculosus (Fv), Fucus serratus (Fs), and Fucus distichus subsp. evanescens (Fe) for their chemical features, endotoxin content, cytotoxicity, and bioactive effects on human outgrowth endothelial cells (OEC) and human mesenchymal stem cells (MSC) as in vitro models for bone function and vascularization. Extracts contained mainly high molecular weight (HMW) fucoidans and were free of endotoxins that may cause inflammation or influence vascularization. OEC tolerated fucoidan concentrations up to 200 µg/mL, and no indication of cytotoxicity was observed. The inflammatory response, however, investigated by real-time PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) and endothelial barrier assessed by impedance measurement differed for the individual extracts. MSC in comparison with endothelial cells were more sensitive to fucoidans and showed partly reduced metabolic activity and proliferation at higher doses of fucoidans. Further results for MSC indicated impaired osteogenic functions in alkaline phosphatase and calcification assays. All tested extracts consistently lowered important molecular mediators involved in angiogenesis, such a VEGF (vascular endothelial growth factor), ANG-1 (angiopoietin 1), and ANG-2 (angiopoietin 2), as indicated by RT-PCR and ELISA. This was associated with antiangiogenic effects at the functional level using selected extracts in co-culture models to mimic bone vascularization processes during bone regeneration or osteosarcoma.     

Tissue architecture and soil fertility controls on decomposer communities and decomposition of roots

Using complementary techniques we observed great differences in decomposition of mono- and dicot roots (Festuca rubra L., red fescue grass and Trifolium pratense L., red clover) in compost and soil of varying fertility which we attribute both to chemical characteristics and differences in tissue architecture. In our study the lignin contents were equal for the root materials while C-to-N ratios and cellulose-to-lignin ratios were higher in fescue roots. Pictures taken with a scanning electron microscope (SEM) showed that the rippled surface of fescue roots appeared largely intact even after prolonged incubation giving scant support to predominantly cocci bacteria, where an angular grid structure on the surface of intact clover roots rapidly yielded. The clover root cavities became deeper over time and seemed a favourable nest for colonising rod shaped bacteria. In addition the nodules on clover roots were abundantly covered by decomposers early on. SEM pictures as well as bands of microbial DNA from polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) on roots degraded in compost indicate that organisms colonising clover and fescue grass roots are inherently different and to a greater extent connected to the root material than the compost. Respiration data showed that soil fertility compared to root material played a smaller, albeit significant role in determining the decomposition pattern. There was no indication of N limitation in decomposition. On the contrary accumulated respiration was highest from infertile soils and had significantly lower T_½ compared to nutrient rich soils, most likely due to a higher maintenance respiration and lower substrate use efficiency of the decomposers. DGGE showed that infertile soils as well as recalcitrant material both led to a larger diversity of decomposers compared to fertile soils or easily degradable materials. All together this study demonstrated a positive relationship between microbial diversity and stress resulting in higher diversity in the recalcitrant the roots and the infertile soils.     

Bilateral renal agenesis in an alpaca cria

A 3-day-old male alpaca cria was presented for lack of vigor and failure to urinate since birth. Based on the history, laboratory data, ultrasonographs, surgical findings, and postmortem examination, the cria was diagnosed with bilateral renal agenesis and hypoplastic bladder, a congenital condition rarely seen in veterinary medicine.     

Association between average daily gain,faecal dry matter content and concentration of Lawsonia intracellularis in faeces

Background

The objective of this study was to investigate the association between average daily gain and the number of Lawsonia intracellularis bacteria in faeces of growing pigs with different levels of diarrhoea.

Methods

A longitudinal field study (n = 150 pigs) was performed in a Danish herd from day 29 to 47 post weaning. Every third day all pigs were weighed, subjected to a clinical examination and faecal samples were obtained. Faecal samples were subjected to dry matter determination and absolute quantification by PCR for L. intracellularis and porcine circovirus type 2 (PCV2). Association between average daily gain, faecal dry matter content, numbers of L. intracellularis bacteria and PCV2 genome copies in faeces was investigated in a multilevel mixed-effects linear model.

Results

Increasing numbers of L. intracellularis log₁₀ bacteria/g faeces were significantly associated with decreasing average daily gain (P < 0.001). The association was decreasing with increasing faecal dry matter content (P < 0.01). The number of PCV2 log₁₀ copies/g faeces was not significantly associated with average daily gain of the pigs (P > 0.5).

Conclusion

The results suggest a potential application of a PCR quantifying L. intracellularis in growing pigs. Faecal dry matter content must be taken into consideration in interpretation of such test results.     

Assessing for genetic and environmental effects on ruminant feed quality in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis>)

Grain samples from a combined intermediate and advanced stage barley breeding trial series, grown at two sites in two consecutive years were assessed for detailed grain quality and ruminant feed quality. The results indicated that there were significant genetic and environmental effects for “feed” traits as measured using grain hardness, acid detergent fibre (ADF), starch and in-sacco dry matter digestibility (ISDMD) assays. In addition, there was strong genotypic discrimination for the regressed feed performance traits, namely Net Energy (NE) and Average Daily Gain (ADG). There was considerable variation in genetic correlations for all traits based on variance from the cultivars used, sites or laboratory processing effects. There was a high level of heritability ranging from 89% to 88% for retention, 60% to 80% for protein and 56% to 68% for ADF. However, there were only low to moderate levels of heritability for the feed traits, with starch 30–39%, ISDMD 55–63%, ADF 56–68%, particle size 47–73%, 31–48% NE and ADG 44–51%. These results suggest that there were real differences in the feed performance of barleys and that selection for cattle feed quality is potentially a viable option for breeding programs.     

C-kit,flt-3, PDGFR-β, and VEGFR2 expression in canine adrenal tumors and correlation with outcome following adrenalectomy

The objective of this retrospective study was to evaluate the expression of receptor tyrosine kinases (RTKs) in canine adrenal tumors and correlate this expression with features of tumor aggressiveness and survival in dogs undergoing adrenalectomy.Forty-three canine adrenal tumors were evaluated for expression of c-kit, fms-like tyrosine kinase 3 (flt-3), platelet-derived growth factor receptor-β (PDGFR-β), and vascular endothelial growth factor receptor 2 (VEGFR2) using immunohistochemistry. Tumor RTK staining characteristics were compared to normal adrenals. Medical records were reviewed for data regarding patient outcome and tumor characteristics. Expression of c-kit, flt-3, PDGFR-β, and VEGFR2 was detected in 26.9%, 92.3%, 96.2%, and 61.5% of cortical tumors and 0%, 63.2%, 47.4%, and 15.8% of pheochromocytomas, respectively. Expression of RTKs was not significantly increased when compared to normal adrenals and did not correlate with survival after adrenalectomy. Receptor tyrosine kinases are not overexpressed in canine adrenal tumors compared to normal adrenal tissue. Therapeutic inhibition of these receptors may still represent an effective approach in cases where receptor activation is present.     

Current distribution of older and deciduous forests as legacies from historical use patterns in a Swedish boreal landscape (1725–2007)

We combine historical maps and satellite derived data to reconstruct the development of a Swedish boreal landscape over the past 300 years. The aim is to understand legacies from past use patterns in present-day forest composition and consequences for conservation objectives from a landscape perspective. We analyze landscape development in cross-tabulation matrixes, building change trajectories. These trajectories are tested in linear models to explain the distribution of present-day landscape composition of coniferous, mixed, and deciduous forests >110 years. Of 49 tested change trajectories, 11 showed a significant association. Associations for mixed and coniferous forests were similar and linked to characteristics such as forest continuity, which characterized the studied landscape. Deciduous older forests did not show any association to forest continuity but were more likely to occur on areas that specifically shifted from forests with grazing in the 1720s to open impediment (likely indicating low tree coverage) in the 1850s. There were large shifts and spatial redistribution in ownerships over time. Use patterns and legacies varied between small- and large-scale ownership categories as well as within small-scale categories. The legacies found in the study indicate a complex origin of heterogeneous landscape elements such as older deciduous forests. Additionally, the origin of the legacies indicates a potential need to diversify conservation management based on the influence of past use patterns. Despite large inconsistencies in historical and contemporary data we argue that this type of analysis could be used to further understand the distribution of landscape elements important for conservation objectives.     

Ontogeny and characterization of blood leukocyte subsets and serum proteins in piglets before and after weaning

Existing knowledge about the development of the porcine immune system was extended by phenotypic characterization of leukocyte subsets and with assessment of Mannan-Binding Lectin (MBL) and immunoglobulin concentrations in peripheral blood of healthy piglets. Single-color and/or double-color flow cytometry using monoclonal antibodies against CD1, CD3, CD4, CD8a CD14, CD21, CD172 (SWC3a), CD284 (TLR4), SLA I, and SLA II were performed to identify T-lymphocyte subsets, B-lymphocytes, monocytes, and granulocytes. ELISA was used to measure the concentration of serum proteins. Several of the analyzed parameters seem to be affected at the time of weaning which took place at 4–5 weeks of age. Using principal component analysis, all analyzed variables – except one – were grouped into 8 factors with distinct developmental profiles. Several of these factors revealed an apparent suppression, a steady state or an acceleration of the developmental age profiles around weaning.In conclusion, results indicate that weaning suppresses a broad spectrum of adaptive immune variables and that this was evident immediately after weaning as well as after a lag period of about one week. On the contrary, variables of the innate immune system seem to be stimulated immediately after weaning. At the time considered to have the highest infection pressure T-cells and TLR4+ cells were markedly enhanced, whereas the expression of SLA I did not seem to be affected by weaning.     

如何在田间评估PCV2疫苗的免疫效果

免疫控制猪圆环病毒2型(Porcine Circovirus 2,PCV2)有时被认为是一项不可思议的工作。然而,免疫预防成功的基础是科学。了解该过程的背景后,在求助于免疫接种时将有助于了解哪些结果是可以预测到的,哪些结果是无法预测到的。     

Reducing the impact of blue mussel (Mytilus edulis) dredging on the ecosystem in shallow water soft bottom areas

• Dredging blue mussels (Mytilus edulis) and thus removing structural elements, inducing resuspension of sediment as well as reducing filtration capacity, will inevitably affect the ecosystem. The study demonstrates that the impacts of fishing can be reduced through gear developments.
• A new light dredge was tested on commercial vessels using two different experimental setups. First, a twin haul experiment tested the standard gear (i.e., a Dutch dredge) against the light dredge by fishing the two gears side by side onboard the same vessel. Second, a single dredge experiment tested the absolute performance of the two gears by fishing in areas with a known blue mussel density.
• Results from the twin haul experiment demonstrate that the weight of sediment retained in the gear per square metre fished is 49% less in the light dredge compared with the Dutch dredge which will reduce resuspension of sediment at the surface. Also, the drag resistance of the light dredge was significantly less (177.1 vs. 202.7 kg m^‐1). In the twin haul experiment no significant difference was found in the catch per unit effort (CPUE) of the two gears. The single dredge experiment, on the other hand, demonstrated a significant increase in CPUE exceeding 200% when using the light dredge.
• Seafloor tracks made by the two dredges could not be distinguished by use of side‐scan sonar and the tracks were still detectable 2 months after fishing.
• It was concluded that replacement of the Dutch dredge with the light dredge would reduce the impact of the fishery on the ecosystem by (i) reducing resuspension of sediment, (ii) reducing fuel consumption, and (iii) potentially reducing energy transfer to the sediment through a reduced gear drag resistance. A potential increase in catch efficiency may reduce the area affected.
• Fishing with the light dredge is discussed in relation to management of Natura 2000 sites.

Copyright © 2014 John Wiley & Sons, Ltd.