Genetic improvement of speed across distance categories in thoroughbred racehorses in Great Britain.

Several studies over recent decades have reported a lack of contemporary improvement in thoroughbred racehorse speed, despite apparent additive genetic variance and putatively strong selection. More recently, it has been shown that some phenotypic improvement is ongoing, but rates are low in general and particularly so over longer distances. Here we used pedigree-based analysis of 692,534 records from 76,960 animals to determine whether these phenotypic trends are underpinned by genetic selection responses, and to evaluate the potential for more rapid improvement. We show that thoroughbred speed in Great Britain is only weakly heritable across sprint (h2 = 0.124), middle-distance (h2 = 0.122) and long-distance races (h2 = 0.074), but that mean predicted breeding values are nonetheless increasing across cohorts born between 1995 and 2012 (and racing from 1997 to 2014). For all three race distance categories, estimated rates of genetic improvement are statistically significant and also greater than can be explained by drift. Taken together our results show genetic improvement for thoroughbred speed is ongoing but slow, likely due to a combination of long generation times and low heritabilities. Additionally, estimates of realised selection intensities raises the possibility that the contemporary selection emerging from the collective actions of horse breeders is weaker than previously assumed, particularly over long distances. We suggest that unmodelled common environment effects may have upwardly biased estimates of heritability, and thus expected selection response, previously.

Racehorses are getting faster.

Previous studies have concluded that thoroughbred racehorse speed is improving very slowly, if at all, despite heritable variation for performance and putatively intensive selective breeding. This has led to the suggestion that racehorses have reached a selection limit. However, previous studies have been limited, focusing only on the winning times of a few elite races run over middle and long distances, and failing to account for potentially confounding factors. Using a much larger dataset covering the full range of race distances and accounting for variation in factors such as ground softness, we show that improvement is, in fact, ongoing for the population as a whole, but driven largely by increasing speed in sprint races. In contrast, speed over middle and long distances, at least at the elite level, appears to be reaching an asymptote. Whether this reflects a selection limit to speed over middle and long distances or a shift in breeding practices to target sprint performances remains to be determined.

Evidence of maternal and paternal age effects on speed in thoroughbred racehorses.

Effects of parental age on offspring viability have been reported in a wide range of species. However, to what extent parental age influences offspring traits beyond viability remains unclear. Moreover, previous research has primarily focused on maternal age effects. The purpose of this study was to test for paternal and maternal age effects on offspring speed in thoroughbred racehorses. We analysed over 900 000 race performances by over 100 000 horses on British racecourses between 1996 and 2019. With knowledge of the age of all 41 107 dams and 2 887 sires at offspring conception, we jointly modelled maternal and paternal age effects using a 'within-individual centring' approach. Within-parents, we identified a significant effect of maternal age on offspring speed of -0.017 yards s-1 yr-1 and a corresponding paternal age effect of -0.011 yards s-1 yr-1. Although maternal age effects were stronger (more negative), the existence and magnitude of paternal effects is particularly noteworthy, given thoroughbred sires have no involvement in parental care. Our results also suggest that the selective disappearance of both sires and dams is ongoing. These findings could potentially be used to optimize thoroughbred racehorse breeding decisions, and more generally, add to the increasing body of evidence that both maternal and paternal age affect a range of offspring characteristics.

Magnetically labeled human natural killer cells, accumulated in vitro by an external magnetic force, are effective against HOS osteosarcoma cells.

We evaluated the efficacy of a novel natural killer (NK) cell delivery system in vitro, and also investigated the antitumor effect of the accumulated cells on HOS osteosarcoma cells. Human peripheral blood mononuclear cells were isolated and co-cultured with inactivated K562 erythroleukaemic cells in the presence of IL-2 for 5 days. CD3- CD56+ NK cells were labeled with immunomagnetic beads and separated using a magnetic cell sorting system. Purity and cytotoxicity against K562 cells and HOS cells of the magnetically labeled NK cells were measured. To evaluate whether magnetically labeled NK cells could be accumulated in a specific area by magnetic force, the NK cells were placed in chamber slides in the presence, or not, of an external magnetic force of a neodymium magnet (diameter: 1.5 mm, height: 3 mm, total magnetic flux density: 0.282 T). Moreover, to investigate the antitumor effect on HOS cells, the magnetically labeled NK cells were added to HOS cells in chamber slides in the presence, or not, of an external magnetic force for various times. HOS cells were subsequently stained with Papanicolaou for histological examination. It was found that the magnetically labeled NK cells were highly purified and had cytotoxicity against target cells. The NK cells were accumulated effectively by the magnetic field and, when the NK cells were added to HOS cells in a chamber slide with a magnet placed beneath, a significantly larger number of HOS cells detached from the magnet zone than from other zones. Apoptosis was detected in most detached HOS cells. In conclusion, these findings indicate that magnetically accumulated NK cells efficiently induced apoptosis in HOS cells, suggesting that magnetic targeting therapy using magnetically labeled NK cells holds promise as an immunotherapy for osteosarcoma.

Characterization of labeled neural progenitor cells for magnetic targeting.

For many diseases and injuries of the central nervous system, transplantation of neural progenitor cells is being evaluated as a possible treatment option. Although local, intravenous and subarachnoid injections have been reported as administration methods of neural progenitor cells, each of these methods has limitations. More effective and minimally invasive cell delivery systems are necessary for transplanting neural progenitor cells. In this study, we have developed a technique to form magnetically labeled neural progenitor cells for a magnetic targeting system. We demonstrated that neural progenitor cells can couple with magnetic beads, and that the labeled neural progenitor cells preserve the characteristics of non-labeled neural progenitor cells, and that they can be localized by magnetic force in vitro. Labeled neural progenitor cells have the potential to be used in magnetic targeting systems in-vivo models.

Hyperbaric oxygen as an adjuvant for athletes.

There has recently been a resurgence in interest in hyperbaric oxygen (HBO) treatment in sports therapy, especially in Japan. Oxygen naturally plays a crucial role in recovery from injury and physiological fatigue. By performing HBO treatment, more oxygen is dissolved in the plasma of the pulmonary vein via the alveolar, increasing the oxygen reaching the peripheral tissues. HBO treatment is therefore expected to improve recovery from injury and fatigue. HBO treatment has been reported to reduce post-injury swelling in animals, and in humans; swelling was also mitigated, but to a lesser extent. Positive results have also been reported regarding tissue remodelling after injury, with injuries involving bones, muscles and ligaments showing improved recovery. Furthermore, HBO treatment has effectively increased recovery from fatigue. This was clearly seen at the Nagano Winter Olympics, where sports players experiencing fatigue were successfully treated, enabling the players to continue performing in the games. Despite its potential, HBO treatment does have its risks. Increasing oxygen levels in tissues poses a risk to DNA through oxidative damage, which can lead to pathological changes in the CNS and the lungs. Regarding the operating of HBO systems, safer administration should be advised. Further research into HBO treatment is required if this therapy is to become more widespread. It should become possible to tailor treatment to an individual's condition in order to use HBO treatment efficiently.

Meniscal regeneration using tissue engineering with a scaffold derived from a rat meniscus and mesenchymal stromal cells derived from rat bone marrow.

The purpose of this study was to regenerate a meniscus using a scaffold from a normal meniscus and mesenchymal stromal cells derived from bone marrow (BM-MSCs). Thirty Sprague-Dawley rat menisci were excised and freeze-thawed three times with liquid nitrogen to kill the original meniscal cells. Bone marrow was aspirated from enhanced green fluorescent protein transgenic Sprague-Dawley rats. BM-MSCs were isolated, cultured for 2 weeks, and 2 x 10(5) cells were then seeded onto the meniscal scaffolds. Using a fluorescent microscope and immunohistochemical staining, repopulation of enhanced green fluorescent protein positive cells was observed in the superficial zone of the scaffold after 1 week of culture, and then in the deep zone after 2 weeks. At 4 weeks, expression of extracellular matrices was detected histologically and expression of mRNA for aggrecan and type X collagen was detected. Stiffness of the cultured tissue, assessed by the indentation stiffness test, had increased significantly after 2 weeks in culture, and approximated the stiffness of a normal meniscus. From this study, we conclude that a scaffold derived from a normal meniscus seeded with BM-MSCs can form a meniscus approximating a normal meniscus.

Flow cytometric discrimination of mesenchymal progenitor cells from bone marrow-adherent cell populations using CD34/44/45(-) and Sca-1(+) markers.

BACKGROUND: Cultured bone marrow adherent cells (BMACs) have been commonly used as stem cells in bone and cartilage regeneration therapy. However, BMACs are actually a heterogeneous cell population, and clinicians might have previously transplanted more fibroblasts or other cells than actual stem cells. The purposes of this study were to (1) isolate immature mesenchymal stem cells with CD34/44/45 and Sca-1 surface-antigen patterns from BMACs using flow-activated cell sorting and (2) investigate their differentiation potential. METHODS: Bone marrow cells were extracted from the mouse femur and cultured. Adherent cells could be identified approximately 3 days after seeding, and nonadherent cells were removed with the medium when it was changed. BMAC samples were cultured for 3, 7, 10, 14, 21, 28, 35, and 42 days after the first seeding. We directly isolated CD34/44/45(-)Sca-1(+) mesenchymal progenitor cells (MPC1) and CD34/45(-)/44(+) Sca-1(+) mesenchymal progenitor cells (MPC2) from BMACs based on their cell surface marker patterns using a fluorescence-activated cell sorter. These subgroups - MPC1, MPC2, and the residual cells in BMACs (non-MPC population: RCs) - were then induced to differentiate into bone, cartilage, and fat using a plate culture. The cultures were examined after histochemical staining on day 14. RESULTS: In a plate culture, the MPC1 population had higher potential to differentiate into osteoblasts, chondrocytes, and lipocytes; whereas MPC2 and RCs differentiated into only two lineages: osteoblasts and lipocytes. The incidence of these multipotential cells was less than 5% among the cultured BMACs. MPC1 proliferated up to 17-fold within 3-4 weeks after separation from floating cells and did not increase thereafter. CONCLUSIONS: BMACs are conventionally thought to differentiate into cartilage only in pellet culture, but we showed that MPC1 produced cartilage-like extracellular matrix in plate culture. MPC1, which are more immature cells than MPC2 and RCs, were multipotential progenitors that showed unique cartilage-differentiation potential. MPC1 had less ability to proliferate in BMAC culture, but they might have higher potential for chondrogenic differentiation.

Telomerase activity in giant cell tumors of bone.

BACKGROUND: A giant cell tumor of bone (GCT) is a histologically benign neoplasma that has an unpredictable pattern of biological aggressiveness. In the present study, we investigated whether there was a correlation between telomere length or the levels of telomerase activity and other clinical features of GCTs, for the possible use of these factors as parameters of aggressiveness or prognosis. METHODS: In 16 surgically resected GCTs specimens, telomere length was assessed by terminal restriction fragments by Southern blot analysis. Telomerase activity was measured by a semiquantitative polymerase chain reaction-based telomeric repeat amplification protocol assay. RESULTS: Telomere length reduction was observed in 69% of the GCT samples. The telomere lengths of tumors were significantly shorter than those of normal tissue (P = .008). The mean telomere length of grade 3 tumors was significantly shorter than those of grade 1 and 2 tumors (P = .038). Telomerase activity was detected in 81% of tumor samples. The level of telomerase activity in tumors with local recurrence was significantly higher than in tumors without local recurrence (P = .011). CONCLUSIONS: These results suggest that telomere length correlates with roentgenographic grade as a result of the frequency of cell division, and high telomerase activity indicates the aggressiveness of GCTs.

Increased apoptosis in human osteoarthritic cartilage corresponds to reduced cell density and expression of caspase-3.

OBJECTIVE: Chondrocyte apoptosis has been described in both human and experimentally induced osteoarthritis (OA), but its importance in the etiopathogenesis of OA is uncertain. The aims of this study were to determine the rate of chondrocyte apoptosis using different methods, and to investigate the relationship between this process and cartilage cellularity, expression of proapoptotic molecules, and expression of antiapoptotic molecules in articular cartilage obtained from patients with OA and from nonarthritic controls. METHODS: We examined the extent of apoptosis in OA and nonarthritic control cartilage using expression of caspase-3, an enzyme that mediates the final stage of cell death by apoptosis, as well as the TUNEL method. We used immunohistochemistry to analyze the expression of a panel of proapoptotic and antiapoptotic molecules that regulate apoptosis in articular cartilage, in order to determine whether the rate of apoptosis is associated with the expression of these molecules. RESULTS: The median (range) percentage of TUNEL-positive chondrocytes in knee OA cartilage (n = 10 specimens), hip OA cartilage (n = 9), and control cartilage (n = 7) was 3.11 (1.67-3.67), 1.86 (1.22-2.89), and 0.39 (0.00-1.78), respectively. When all cartilage samples were pooled, apoptosis showed a strong inverse correlation with cellularity (r = -0.74, P < 0.0001). The percentage (range) of cells expressing caspase-3 in the 3 groups was 15.70 (7.40-20.50), 15.77 (7.42-20.5), and 7.40 (5.90-8.00), respectively. One-way analysis of variance showed that the differences between groups for both TUNEL-positive cells and expression of caspase-3 were statistically significant (P < 0.0001). There was a significant positive correlation between TUNEL-positive cells and expression of caspase-3 (r = 0.654, P< 0.01). CONCLUSION: The data suggest that apoptosis is increased, on average, 2-4-fold in OA cartilage. Considering that OA develops over many years, such an increase in the rate of apoptosis in the articular cartilage could play an important role in the disease process.