Forage legume silage and cold-pressed rapeseed cake for dairy bull calves

Rearing of organic dairy bull calves can be contradictory, using calves with a high need for quality protein at the same time as a high intake of forage (>60% up to 6 months of age) is required. Forage legumes contain more crude protein than grass, but with high rumen degradability of the protein. When legume forage is fed together with energy-rich cold-pressed rapeseed cake (CRC) the protein in the feeds can be utilized to a higher extent and a satisfactory calf weight gain can be achieved as shown. Using locally produced protein feeds instead of the often used soya bean meal (SBM) is of great interest not only in organic but also in conventional feeding because of environmentally advantages

Influence of slaughter age and carcass suspension on meat quality in Angus heifers

This study investigated the effects of pelvic suspension and slaughter age on longissimus thoracis et lumborum (LTL) from 40 heifers with at least 75% Angus breeding. A total of 20 heifers were slaughtered directly from pasture at 18 months of age, and carcass sides were hung either by the Achilles tendon or the pelvic bone. The other 20 heifers were assigned to an additional winter housing period and slaughtered at 22 months of age; carcass sides were hung only by Achilles suspension. All carcasses were electrically stimulated and assessed according to the EUROP carcass classification system. In addition, the LTL muscles were aged for 7 or 14 days before meat quality was evaluated for intramuscular fat (IMF), drip loss, colour, shear force, compression and sensory analysis. The 22-month-old heifers were heavier, fatter and had more IMF than 18-month-old heifers. Conformation scores (muscling) did not differ between the two slaughter groups. Pelvic suspension reduced both between- and within-animal variation for peak force, total energy and compression peak force. For the 18-month-old heifers, pelvic suspension also decreased peak force, total energy and compression variables for the LTL muscles from both ageing periods, whereas Achilles-suspended samples had lower shear force values only at day 14. Sensory analysis showed that pelvic-suspended sides had greater tenderness, lower bite resistance, less threadiness, higher juiciness and meat flavour and less visible marbling than meat from Achilles-suspended sides. Pelvic-suspended sides at 18 months of age were similar in peak force and total energy values to the 22-month-old heifers. The importance of ageing the Achilles-suspended sides was more obvious for samples from 18-month-old heifers than from the 22-month-old animals. The correlations between the different instrumental measurements and sensory tenderness were considerably higher for carcasses suspended by the Achilles tendon (r = -520.55 to 0.20) than for those hung by the pelvic bone (r = -520.25 to 0.19). More correlations between sensory-evaluated tenderness and shear variables were significant after 7 days (n = 6) of ageing than after 14 days (n = 4) of ageing. This study clearly shows the benefits of pelvic suspension, which reduces the need for additional feeding after pasture

The skeletal phenotype of chondroadherin deficient mice

Chondroadherin, a leucine rich repeat extracellular matrix protein with functions in cell to matrix interactions, binds cells via their a2b1 integrin as well as via cell surface proteoglycans, providing for different sets of signals to the cell. Additionally, the protein acts as an anchor to the matrix by binding tightly to collagens type I and II as well as type VI. We generated mice with inactivated chondroadherin gene to provide integrated studies of the role of the protein. The null mice presented distinct phenotypes with affected cartilage as well as bone. At 3–6 weeks of age the epiphyseal growth plate was widened most pronounced in the proliferative zone. The proteome of the femoral head articular cartilage at 4 months of age showed some distinct differences, with increased deposition of cartilage intermediate layer protein 1 and fibronectin in the chondroadherin deficient mice, more pronounced in the female. Other proteins show decreased levels in the deficient mice, particularly pronounced for matrilin-1, thrombospondin-1 and notably the members of the a1-antitrypsin family of proteinase inhibitors as well as for a member of the bone morphogenetic protein growth factor family. Thus, cartilage homeostasis is distinctly altered. The bone phenotype was expressed in several ways. The number of bone sialoprotein mRNA expressing cells in the proximal tibial metaphysic was decreased and the osteoid surface was increased possibly indicating a change in mineral metabolism. Micro-CT revealed lower cortical thickness and increased structure model index, i.e. the amount of plates and rods composing the bone trabeculas. The structural changes were paralleled by loss of function, where the null mice showed lower femoral neck failure load and tibial strength during mechanical testing at 4 months of age. The skeletal phenotype points at a role for chondroadherin in both bone and cartilage homeostasis, however, without leading to altered longitudinal growth

Extracellular ATP released by osteoblasts is a key local inhibitor of bone mineralisation

Previous studies have shown that exogenous ATP (>1µM) prevents bone formation in vitro by blocking mineralisation of the collagenous matrix. This effect is thought to be mediated via both P2 receptor-dependent pathways and a receptor-independent mechanism (hydrolysis of ATP to produce the mineralisation inhibitor pyrophosphate, PPi). Osteoblasts are also known to release ATP constitutively. To determine whether this endogenous ATP might exert significant biological effects, bone-forming primary rat osteoblasts were cultured with 0.5-2.5U/ml apyrase (which sequentially hydrolyses ATP to ADP to AMP + 2Pi). Addition of 0.5U/ml apyrase to osteoblast culture medium degraded extracellular ATP to <1% of control levels within 2 minutes; continuous exposure to apyrase maintained this inhibition for up to 14 days. Apyrase treatment for the first 72 hours of culture caused small decreases (≤25%) in osteoblast number, suggesting a role for endogenous ATP in stimulating cell proliferation. Continuous apyrase treatment for 14 days (≥0.5U/ml) increased mineralisation of bone nodules by up to 3-fold. Increases in bone mineralisation were also seen when osteoblasts were cultured with the ATP release inhibitors, NEM and brefeldin A, as well as with P2X1 and P2X7 receptor antagonists. Apyrase decreased alkaline phosphatase (TNAP) activity by up to 60%, whilst increasing the activity of the PPi-generating ecto-nucleotide pyrophosphatase/phosphodiesterases (NPPs) up to 2.7-fold. Both collagen production and adipocyte formation were unaffected. These data suggest that nucleotides released by osteoblasts in bone could act locally, via multiple mechanisms, to limit mineralisation

A cell-autonomous requirement for neutral sphingomyelinase 2 in bone mineralization

nSMase2, which cleaves sphingomyelin to generate bioactive lipids, is required for chondrocyte apoptosis and, cell autonomously, for bone mineralization

The Appearance and Modulation of Osteocyte Marker Expression during Calcification of Vascular Smooth Muscle Cells

Vascular calcification is an indicator of elevated cardiovascular risk. Vascular smooth muscle cells (VSMCs), the predominant cell type involved in medial vascular calcification, can undergo phenotypic transition to both osteoblastic and chondrocytic cells within a calcifying environment.In the present study, using in vitro VSMC calcification studies in conjunction with ex vivo analyses of a mouse model of medial calcification, we show that vascular calcification is also associated with the expression of osteocyte phenotype markers. As controls, the terminal differentiation of murine calvarial osteoblasts into osteocytes was induced in vitro in the presence of calcifying medium (containing ß-glycerophosphate and ascorbic acid), as determined by increased expression of the osteocyte markers DMP-1, E11 and sclerostin. Culture of murine aortic VSMCs under identical conditions confirmed that the calcification of these cells can also be induced in similar calcifying medium. Calcified VSMCs had increased alkaline phosphatase activity and PiT-1 expression, which are recognized markers of vascular calcification. Expression of DMP-1, E11 and sclerostin was up-regulated during VSMC calcification in vitro. Increased protein expression of E11, an early osteocyte marker, and sclerostin, expressed by more mature osteocytes was also observed in the calcified media of Enpp1(-/-) mouse aortic tissue.This study has demonstrated the up-regulation of key osteocytic molecules during the vascular calcification process. A fuller understanding of the functional role of osteocyte formation and specifically sclerostin and E11 expression in the vascular calcification process may identify novel potential therapeutic strategies for clinical intervention

Alkaline Phosphatases: Structure, substrate specificity and functional relatedness to other members of a large superfamily of enzymes

Our knowledge of the structure and function of alkaline phosphatases has increased greatly in recent years. The crystal structure of the human placental isozyme has enabled us to probe salient features of the mammalian enzymes that differ from those of the bacterial enzymes. The availability of knockout mice deficient in each of the murine alkaline phosphatase isozymes has also given deep insights into their in vivo role. This has been particularly true for probing the biological role of bone alkaline phosphatase during skeletal mineralization. Due to space constraints this mini-review focuses exclusively on structural and functional features of mammalian alkaline phosphatases as identified by crystallography and probed by site-directed mutagenesis and kinetic analysis. An emphasis is also placed on the substrate specificity of alkaline phosphatases, their catalytic properties as phosphohydrolases as well as phosphodiesterases and their structural and functional relatedness to a large superfamily of enzymes that includes nucleotide pyrophosphatase/phosphodiesterase