Comparing skeletal development of wild and hatchery-reared Senegalese sole (Solea senegalensis, Kaup 1858): evaluation in larval and postlarval stages

The Senegalese sole is a marine pleuronectiform that naturally occurs in Southern Europe and Mediterranean region where it is being produced in aquaculture, in particular in Portugal and Spain. The aim of this study was to assess the quality of hatchery-reared larvae in comparison with those reared in the wild, and determine to which extension wild growing larvae are also affected by skeletal deformities. The main structures affected included those forming the axial skeleton, the caudal fin complex and both anal and dorsal fins, with the most prevalent anomalies affecting caudal vertebrae and arches. Hatchery-reared fish presented a higher incidence of deformities (79%) compared with the 19% observed in wild specimens. In wild postlarvae collected in Autumn no deformities were observed. This work clearly shows that wild Senegalese sole present less skeletal deformities than those hatchery-reared during larval stages, indicating a selective mortality of wild deformed fish and/or an effect of aquaculture-related rearing conditions in the development of skeletal deformities in sole.     

Decellularization of Extracellular Matrix from Equine Skeletal Muscle

Equine represents an attractive animal model for musculoskeletal tissue diseases, exhibiting much similarity to the injuries that occur in humans. Cell therapy and tissue bioengineering have been widely used as a therapeutic alternative by regenerative medicine in musculoskeletal diseases. Thus, the aim of this study was to produce an acellular biomaterial of equine skeletal muscle and to evaluate its effectiveness in supporting the in vitro culture of equine induced pluripotency stem cells (iPSCs). Biceps femoris samples were frozen at −20°C for 4 days and incubated in 1% sodium dodecyl sulfate (SDS), 5 mM EDTA + 50 mM Tris and 1% Triton X-100; the effectiveness of the decellularization was evaluated by the absence of remnant nuclei (histological and 4′,6-diamidino-2-phenylindole [DAPI] analysis), preservation of extracellular matrix (ECM) proteins (immunofluorescence and immunohistochemistry) and organization of ECM ultrastructure (scanning electron microscopy). Decellularized samples were recellularized with iPSCs at the concentration of 50,000 cells/cm² and cultured in vitro for 9 days, and the presence of the cells in the biomaterial was evaluated by histological analysis and presence of nuclei. Decellularized biomaterial showed absence of remnant nuclei and muscle fibers, as well as the preservation of ECM architecture, vascular network and proteins, laminin, fibronectin, elastin, collagen III and IV. After cellularization, iPSC nuclei were present at 9 days after incubation, indicating the decellularized biomaterial-supported iPSC survival. It is concluded that the ECM biomaterial produced from the decellularized equine skeletal muscle has potential for iPSC adhesion, representing a promising biomaterial for regenerative medicine in the therapy of musculoskeletal diseases.     

Adaptation characteristics of mitochondria in plateau pikas at different altitudes

AIM: To investigate the adaptive characteristics of mitochondria in plateau pikas at different altitudes. METHODS: According to the altitudes of the capture area, plateau pikas were divided into 4 300 m group (Mado Star Sea, n=6) and 2 900 m group (Laoji Mountain South, n=6). The red blood cells and hemoglobin of the animals in 2 groups were measured. The levels of mitochondrial oxidative phosphorylation (OXPHOS) and fatty acid oxidation (FAO) in the liver and skeletal muscle, and mitochondrial H₂O₂ production level in the skeletal muscle were determined by high-resolution mitochondrial respirometer. RESULTS: The hemoglobin of the plateau pika in 4 300 m group was significantly lower than those in 2 900 m group. The liver state 4 respiration rate of mitochondrial respiratory chain complex I (CI-LEAK), mitochondrial respiratory chain complex I state 3 respiration rate (CI-OXPHOS) and respiratory chain complex I+Ⅱ state 3 respiration rate(CI+Ⅱ-OXPHOS), electron transfer capacity of respiratory chain complex I+Ⅱ (CI+Ⅱ-ETS), the electron transport capacity of the respiratory chain complex Ⅱ (CⅡ-ETS) in 2 900 m group were increased, and the coupling efficiency was decreased compared with 4 300 m group (P<0.05). The skeletal muscle CI-LEAK,CI+Ⅱ-OXPHOS and CI+Ⅱ-ETS were higher than those in 4 300 m group. Mitochondrial state 4 respiration rate with palmitic acid carnitine and malate as substrates (FAO-LEAK) of liver and skeletal muscle in 2 900 m group were increased compared with 4 300 m group. The skeletal muscle with palmitic acid carnitine as a substrate for fatty acid metabolism state 3 respiratory rate (FAO-OXPHOS) was increased in 2 900 m group compared with 4 300 m group. The H₂O₂/O₂ flow ratio (ΔH₂O₂) in 2 900 m group was lower in state 4 respiration rate of mitochondrial respiratory chain complex I (LEAK), state 3 of mitochondrial respiratory chain complex I (OXPHOS), state 3 of mitochondrial respiratory chain complex I+Ⅱ (OXPHOS*) and electron transfer capacity of respiratory chain complex I+Ⅱ (ETS) than 4 300 m group. CONCLUSION: Plateau pikas with different altitudes have different mitochondrial adaptation characteristics.     

Effects of Water Extract from Different Parts of Betelnut on Physiological Indexes in Mice

The experiment was aimed to explore the influence of water extract from fresh betelnut's different parts on physiological indexes in mice.Four-week-old and seven-week-old SPF KM mice at the weight of (18±2)g and (35±2)g were used in the experiment,respectively.48 four-week-old mice with half male and half female were randomly divided into four groups.These mice were intragastricly administered with distilled water (control group,CK),water extract from betelnut,water extract from fresh betelnut's peels and seeds once a day and were observed for 14 days to determine the survival rate,feed intake,water intake and body temperature.36 seven-week-old male mice were randomly divided into 4 groups to determine deformity rate of sperm in mice,and the intragastric administration was the same as above.The results showed that the survival rate of betelnut's seeds were 0 at the sixth day,and other treatments had no death;Compared with CK,the experimental treatments' feed intake and water intake were reduced,and they were the lowest in the betelnut's seeds treatment and had significant difference with other treatments (P<0.05),the betelnut treatment took the second place.All the treatments' body temperature were lower than CK,and the peels treatment's body temperature was the most stable one and was similar to CK(P>0.05),the body temperature in the betelnut's seeds treatment was the lowest and had significant difference with other treatments (P<0.05).Compared to CK,the other 3 treatments all increased sperm deformity rate of mice,the highest sperm deformity rate went to betelnut's seeds treatment (P<0.01),and betelnut treatment took the second place (P<0.01),then was the peels treatment (P<0.05),moreover,there were extremely significant difference among these 3 treatments (P<0.01).The results showed that the water extract from fresh betelnut's seeds had the greatest influence on mice,then was the water extract from betelnut,while the impact of water extract from fresh betelnut's peels was less.     

Effect of acupuncture on mitophagy of skeletal muscle in rats with exercise-induced skeletal muscle damage

AIM: The effect of acupuncture on mitophagy-related protein expression in skeletal muscle of rats after heavy-load exercise was investigated to explore the role of acupuncture in the repairment of exercise-induced skeletal muscle damage. METHODS: Male adult Sprague-Dawley rats (n=128) were randomly divided into 4 groups:control (C, n=8) group, exercise (E, n=40) group, acupuncture (A, n=40) group, and exercise and acupuncture (EA, n=40) group. The rats in E group and EA group performed an eccentric exercise, and the rats in A group and EA group immediately after exercise received acupuncture treatment. The rats in the latter 3 groups were further divided into 0 h, 12 h, 24 h, 48 h and 72 h sub-groups (n=8), and soleus muscle was collected at each time point. The transmission electron microscopy was used to observe the ultrastructural changes of the mitochondria in skeletal muscle. The content of citrate synthase (CS) was measured by ELISA. The protein expression of skeletal muscle PTEN-induced putative kinase 1 (PINK1), parkin and microtubule-associated protein 1 light chain 3 (LC3) was determined by Western blot. RESULTS: After the heavy-load exercise, the mitochondria swelled and accumulated under cell membrane. The number of mitophagosomes was increased, and the content of CS was significantly decreased (P<0.05). The expression of PINK1, parkin and LC3 was significantly elevated (P<0.05). However, the acupuncture intervention after exercise promoted the recovery of mitochondrial ultrastructure, attenuated mitophagolysosome formation, maintained CS content and down-regulated the expression of PINK1, parkin and LC3 (P<0.05). CONCLUSION: Heavy-load exercise causes the damages of mitochondrial structure and function in the skeletal muscle and activates PINK1/parkin pathway to induce excessive occurrence of mitophagy. Acupuncture intervention after exercise is able to alleviate the damage of mitochondria in the skeletal muscle through decreasing the expression of mitochondrial outer membrane protein PINK1, reducing the recruitment of downstream cytoplasmic protein parkin, thereby affecting the combination of LC3 and mitochondria to inhibit the overactivation of mitophagy.     

Farriery for the foal: A review part 1: Basic trimming

The extensive nature of this topic warrants this review paper to be divided into two parts: ‘Routine basic trimming in foals’ and ‘Therapeutic farriery in foals’. Hoof care in the first few months of life is serious business and should never be taken lightly. Good farriery is vital for the development of the hoof capsule and the conformation of the limb. Management of the feet and limbs during the juvenile period will often dictate the success of the foal as a sales yearling or mature sound athlete. Overall hoof care of a foal is often a joint venture between the veterinarian and the farrier. Part 1 of this paper will outline the concept of a footcare programme, examination of the foal's feet and limbs along with what constitutes good basic farriery to properly apply the trim. It will also emphasise the importance of maintaining a good veterinarian-farrier relationship - the farrier being responsible for basic trimming with veterinary oversight when necessary, and if orthopaedic disorders develop, the farrier will have significant input with therapeutic farriery.     

饲料磷水平对草鱼形体的影响及其骨骼畸形的判定

本试验旨在研究饲料中不同磷水平对草鱼形体的影响,并对其骨骼畸形进行判定.选取初始均重为22.29 g的草鱼270尾,随机分为6个组,每组设3个重复,每个重复放鱼15尾,分别投喂合0、13、29、45、61、77 g/kg磷酸二氢钙的纯化饲料.试验期为75 d.结果表明:饲料中磷酸二氢钙添加量为29 g/kg的组草鱼体长...     

应用酵母双杂交技术筛选猪Calsarcin-2蛋白结合蛋白

Calsarcins是钙调磷酸酶肌小节特异性结合蛋白家族,本实验旨在研究Calsarcin-2(CS2)在猪骨骼肌纤维形成和信号通路调控中的功能。应用酵母双杂交方法筛选猪骨骼肌组织中CS2蛋白结合蛋白,从猪骨骼肌文库中筛选出180个阳性克隆,经分析得到12个相互作用的蛋白,研究发现骨骼肌α-肌动蛋白1(ACTA1),肌联蛋白(TTN)及钙调素1(CALM1)为CS2互作蛋白。ACTA1参与肌肉收缩过程,TTN调节肌小节动力传导及Z线装配,CALM1参与钙离子信号传导。结果表明,CS2在维持Z线结构稳定和参与钙离子信号传导方面起重要作用。