In Vitro Expression of the Extracellular Matrix Components Aggrecan,Collagen Types I and II by Articular Cartilage‐Derived Chondrocytes

The extracellular matrix (ECM) of hyaline cartilage is perfectly suited to transmit articular pressure load to the subchondral bone. Pressure is transferred by a high amount of aggrecan‐based proteoglycans and collagen type II fibres in particular. After any injury, the hyaline cartilage is replaced by fibrocartilage, which is low in proteoglycans and contains collagen type I predominantly. Until now, long‐term results of therapeutic procedures including cell‐based therapies like autologous chondrocyte transplantation (ACT) lead to a replacement tissue meeting the composition of fibrocartilage. Therefore, it is of particular interest to discover how and to what extent isolation and in vitro cultivation of chondrocytes affect the cells and their expression of ECM components. Hyaline cartilage‐derived chondrocytes were cultivated in vitro and observed microscopically over a time period of 35 days. The expression of collagen type I, collagen type II and aggrecan was analysed using RT‐qPCR and Western blot at several days of cultivation. Chondrocytes presented a longitudinal shape for the entire cultivation period. While expression of collagen type I prevailed within the first days, only prolonged cultivation led to an increase in collagen type II and aggrecan expression. The results indicate that chondrocyte isolation and in vitro cultivation lead to a dedifferentiation at least to the stage of chondroprogenitor cells.     

Infection of the enteric nervous system by Borna disease virus (BDV) upregulates expression of Calbindin D-28k

Borna disease virus (BDV) is a neurotropic agent infecting distinct neuronal subpopulations in the central nervous system of various mammalian species possibly including humans. Horses, a major natural host for BDV, show gastrointestinal dysfunctions besides characteristic neurological symptoms. Therefore, we hypothesized that enteric neurons may be targets of BDV replication. The presence of BDV-specific antigen in subpopulations of the ENS was investigated. Four-week-old Lewis rats were infected intracerebrally and sacrificed 4-14 weeks post infection (p.i.). BDV-immunoreactive neurons were found in submucous and myenteric neurons of the proximal colon. Fourteen weeks p.i., the proportion of BDV-positive neurons was 44+/-17 and 24+/-7% in the submucous and myenteric plexus, respectively. The majority of BDV-positive myenteric neurons showed immunoreactivity for choline acetyltransferase. Expression of Calbindin D-28k (CALB) was found in 96% of submucous and 67% of myenteric BDV-immunoreactive neurons. Additionally, the number of CALB-immunoreactive neurons was significantly higher in the myenteric plexus of infected rats compared to controls. These data indicate that BDV infects specific subpopulations of enteric neurons. Therefore, the ENS might serve as a site for BDV replication and as an immunoprivileged reservoir for BDV. In addition, upregulation of CALB in neurons of the myenteric plexus is probably induced during BDV-infection.     

Modelling Discharge and Sediment Yield at Catchment Scale Using Connectivity Components

Knowledge about connectivity and what affects it, through space and time, is needed for taking appropriate action at the right place and/or time by stakeholders. Various conceptual frameworks for hydrological and sediment connectivity have been developed in recent years. For most of these frameworks, the objective was to conceptualise connectivity, not necessarily to infer it from measurements. Studies focussing on measurements of connectivity have so far not been done often. Because of lack of data on connectivity, few real‐world data have been used in recent connectivity modelling studies. The aim of this study was to demonstrate that existing data can be used to assess governing factors of connectivity, and how these change over time. Data from three catchments in Navarre, Northern Spain, were used to assess factors that influence hydrologic and sediment connectivity. These factors were used as components in a linear model for discharge and suspended‐sediment yield. Three components of connectivity were distinguished: topographical, biological and soil. Changes in the topographical component for the studied periods were considered relatively small, and, therefore, kept constant. Changes in the biological component were determined using the Normalised Difference Vegetation Index. Changes in the soil component were assessed using an Antecedent Precipitation Index. Nash–Sutcliffe model efficiency coefficients were between 0·49 through 0·62 for the discharge models and between 0·23 through 0·3 for the sediment‐yield models. We recommend applying the model at smaller spatial scales than catchment scale to minimise the lumping of spatial variability in the components. Copyright © 2016 John Wiley & Sons, Ltd.     

Management effects on forms of phosphorus in soil and leaching losses

We should know the effects of soil use and management on the contents and forms of soil phosphorus (P) and the resulting potential for leaching losses of P to prevent eutrophication of surface water. We determined P test values, amounts of sequentially extracted forms of P, P sorption capacities and degrees of P saturation in 20 differently treated soils and compared these data with leaching losses in lysimeters. One-way analyses of variance indicated that most fractions of P were significantly influenced by soil texture, land use (grassland, arable or fallow or reafforestation), mineral fertilization and intensity of soil management. Generally, sandy soils under grass and given large amounts of P fertilizer contained the most labile P and showed the largest P test values. Fallow and reafforestation led to smallest labile P fractions and relative increases of P extractable by H₂SO₄ and residual P. Arable soils with organic and mineral P fertilization given to crop rotations had the largest amounts of total P, labile P fractions and P test values. The mean annual concentrations of P in the lysimeter leachates varied from 0 to 0.81 mg l^–1 (mean 0.16 mg l^–1) and the corresponding leaching losses of P from < 0.01 to 3.2 kg ha^–1 year^–1 (mean 0.3 kg P ha^–1 year^–1). These two sets of data were correlated and a significant exponential function (R² = 0.676) described this relation. Different soil textures, land uses and management practices resulted in similar values for P leaching losses as those for the amounts of labile P fractions. Surprisingly, larger rates of mineral P fertilizer did not necessarily result in greater leaching losses. The contents of P extracted by NaHCO₃ and acid oxalate and the degrees of P saturation were positively correlated with the concentrations of P in leachates and leaching losses. As the P sorption capacity and degree of P saturation predicted leaching losses of P better than did routinely determined soil P tests, they possibly can be developed as novel P tests that meet the requirements of plant nutrition and of water protection.     

Estimating the effects of set-aside on water quality: scaling-up of lysimeter studies

Since 1990, agriculture in the five new federal states of Germany has experienced a fundamental structural change. As much as 10 per cent of the 6·2 million ha of previously intensively farmed agricultural land were set-aside abruptly. In the spring of 1991, a lysimeter trial (filled with soils common in the catchment area of the Elbe River), was set up to investigate the impact of set-aside on the water and solute balance. The studies proved that restricting agricultural use in areas previously farmed intensively by converting them into permanent or rotation fallow will result in measurable changes in deep percolation (ground-water recharge) and water quality in less than one year. The results of the lysimeter studies were extrapolated to calculate the effects of set-aside in a catchment area (about 2 500 ha) with similar meteorological and soil conditions. The calculations showed that increasing the area under rotation fallow from 8 to 15 per cent increases the nitrogen load of the stream draining the catchment by about 5 per cent. Copyright © 1999 John Wiley & Sons, Ltd.     

New aspects in the pathogenesis of abomasal displacement

Impaired abomasal motility and an increased accumulation of gas are prerequisites for displacement of the abomasum in the cow. Predisposing factors are the breed (e.g. Holstein-Friesian, Simmental-Red-Holstein cross breeds and Guernsey), genetic background, twin pregnancy, first weeks of lactation, metabolic disorders (ketosis, increased lipomobilisation, insulin resistance), high-concentrate and low-fibre diets, as well as other concomitant diseases, such as endometritis, mastitis and claw disorders. There does not appear to be a strong correlation between increased milk yield or endotoxaemia and abomasal displacement. Recent studies have focused on possible functional disorders of the enteric nervous system within the abomasal wall, since cattle with abomasal displacement have an increased activity of neuronal nitric oxide synthase, as well as decreased acetylcholine sensitivity. In addition, there appear to be significant differences between breeds in the levels of the neurotransmitters substance P (SP) and vasoactive intestinal peptide (VIP) in the abomasal wall. For example, SP (stimulatory) was significantly less in German Holsteins in comparison to the German Fleckvieh, whereas VIP (inhibitory) was markedly increased. These risk factors may explain why Holstein cows are more susceptible to abomasal displacement than other breeds.