Estimation of bone mineral density and breaking strength of laying hens based on scans of computed tomography for body composition analysis

1. The objective of this study was to evaluate the prediction potential of a computer tomography (CT) data collection protocol for determining total body composition used for analysis of tibiotarsal bone quality features.

2. The CT image acquisition was performed on 54 healthy TETRA SL genotype laying hens at 90 weeks of age as well as in the 69th week of the egg production period in vivo and their tibiotarsal bones, ex vivo.

3. Breaking strengths and ash content of the tibiotarsal bones were estimated based on the calculated mineral density of skeletal and tibiotarsal bones by means of CT with an estimation accuracy R² 0.963 and 0.975, respectively.

4. In conclusion, the current investigation demonstrated that the acquisition protocol of CT for total-body composition analysis has a good potential for measuring the mineral status and breaking strength of the reference bone in laying hen.     

In vitro selective suppression of feline myeloid colony formation is attributable to molecularly cloned strain of feline leukemia virus with unique long terminal repeat

Molecularly cloned feline leukemia virus (FeLV)-clone 33 (C-33), derived from a cat with acute myelocytic leukemia (AML), was examined to assess its relation to the pathogenesis of AML and myelodysplastic syndrome (MDS). To evaluate in vitro pathogenicity of FeLV C-33, bone marrow colony-forming assay was performed on marrow cells infected with FeLV C-33 or an FeLV subgroup A strain (61E, a molecularly cloned strain with minimal pathogenicity). The myeloid colony-forming activity of feline bone marrow mononuclear cells infected with FeLV C-33 was significantly lower than that of cells infected with 61E. This suggests that FeLV C-33 has myeloid lineage-specific pathogenicity for cats, and that FeLV C-33 infection is useful as an experimental model for investigating pathogenesis of MDS and AML.     

鸡骨髓内单核细胞,巨噬细胞和多核巨细胞发生的形态特点

对雏鸡骨髓内单核细胞，巨噬细胞和多核巨细胞发生的形态结构进行了光镜和电镜观察，从幼稚阶段的单核细胞开始，细胞表面出现突起，从原始阶段开始胞质内出现颗粒。随着细胞向成熟阶段发展，突起和颗粒的数量增多，巨噬细胞的形态、结构与幼稚单核细胞相近。     

What is your diagnosis? Shoulder mass in a dog with lameness

Abstract: An adult castrated male Golden Retriever of unknown age was presented with a history of weight loss and progressive left thoracic limb lameness. On physical examination, a solid mass was palpated on the left scapula that had areas of lysis on radiographs and an area of cortical bone loss on ultrasound. Hepatomegaly, abdominal distension, and numerous intra‐abdominal soft tissue masses were also found. Fine‐needle aspirates of the scapula and several abdominal masses contained numerous free nuclei mixed with fewer individualized, intact cells that were round in shape and rarely formed small sheets. The cells had high nuclear to cytoplasmic ratios, central nuclei, coarsely stippled chromatin, 1–2 prominent nucleoli, and basophilic cytoplasm with indistinct cell borders. The cytopathologic interpretation was neuroendocrine neoplasia, either metastatic or multicentric. The dog was subsequently euthanized and based on gross and histologic findings at necropsy, a diagnosis of pheochromocytoma with multiple metastases was made. The neoplastic cells stained positive with Grimelius stain and were immunoreactive for synaptophysin and chromogranin A. Pheochromocytomas are rare tumors in dogs and uncommonly undergo distant metastasis, especially to bone.     

Determinants of bone mass,density and growth in growing dogs with normal and osteopenic bones

Survey radiographs of all the growing dogs aged up to 6 months, which were presented to the IVRI polyclinics during the 10 year period were screened to study the determinants of bone mass, density and growth. On the basis of clinical history and radiographic evaluation of long bones, the cases were categorized as normal or osteopenic. The relative cortical density (RCD), cortical index (CI), diameter of bone at the distal metaphysis (DDFM) and the width of the growth plate (WFGP) were determined by taking the femur as a model bone in German shepherd, Doberman and Spitz breeds of dogs at different age groups. The results showed that the RCD was the least in 0–2 month old normal growing dogs in all the breeds. As the age advanced up to 6 months the RCD increased 20–25%, and at 6 months, Spitz and Doberman showed significant increase (P < 0.05) in the RCD. In osteopenic bones, RCD remained less (25–50%) than that of normal animals at all age groups, and at 2–6 months of age, RCD in osteopenic bones was significantly lesser than in normal animals in GSD and Spitz breeds. The CI was also the least at 0–2 months of age in normal dogs. The CI increased about 50% at 4–6 months of age in GSD and Spitz. Whereas in Dob., there was no appreciable change in the CI at different age groups, and at 2–6 months it was significantly (P < 0.05) lesser than that of Spitz. In osteopenic bones, the CI was 25–75% lesser than that of normal animals at different age groups, and at 4–6 months there was significant difference (P < 0.05) between the normal and osteopenic bones in GSD and Spitz. The DDFM was the least in 0–2 month old normal growing dogs, and as the age advanced, it increased 10–20% up to 6 months. However, no significant difference in the DDFM was seen between breeds and also between the normal and osteopenic bones at different age groups. In normal animals, the WFGP was highest in the early age, subsequently it reduced 50–75% and at 4–6 months there was significant decrease (P < 0.05) in all the breeds of dogs. And at 4–6 months, there was significant (P < 0.05) difference in the WFGP between breeds, it was the least in Spitz and maximum in Dob., suggesting faster growth plate closure in Spitz than in GSD and Dob. breeds. In osteopenic bones, WFGP was generally more than in normal animals, and at 4–6 months (about 3–5 times more) there was significant difference (P < 0.05) between the normal and osteopenic bones in all breeds, indicating that physeal closure may be delayed in osteopenic bones. The results indicate that among different breeds Doberman breed has the least bone mass and may be more prone to osteopenia; whereas Spitz has the strongest bone.     

Effects of gelatin prepared from calf bones rich in phosphorus on broiler performance,bone characteristics and digestive enzymes activity

1. Gelatin prepared from calf bones (GCB) is a novel source of high-quality protein and phosphorus. Its inclusion in broiler chicken diets may improve bone strength, plasma and digestive alkaline phosphatase activity (ALP), phosphorus digestibility and performance of broilers. Therefore, di-calcium phosphate in a corn-soy control diet was replaced with 12, 24, and 36 g/kg of GCB in a completely randomised design with four treatments of six replicates and 10 chicks in each pen. The trial lasted from 1 to 42 d of age.

2. Body weight and feed intake were measured weekly. Plasma calcium and phosphorus concentration along with plasma and digestive ALP were assayed throughout the trial. Trypsin, α-amylase, lipase and total protease activity were assayed at 14 and 28 d of age. Tibia ash, calcium and phosphorus content and breaking strength were measured at 14, 28 and 42 d of age. Phosphorus digestibility was measured at 36 d of age.

3. Body weight and feed intake showed no significant differences between controls and diets containing 12 and 36 g/kg GCB. Tibia ash and tibia length were increased by supplementation of GCB (P ≤ 0.001). Tibia calcium and phosphorus content were increased by GCB inclusion at 14 d of age (P ≤ 0.001). Digestive alkaline phosphatase activity was increased and trypsin activity was reduced by inclusion of GCB (P ≤ 0.001; P ≤ 0.004). α-amylase activity decreased by inclusion of 12 and 24 g/kg GCB, whereas an increase in α-amylase activity was observed by inclusion of 36 g/kg GCB (P ≤ 0.001). Supplementation of diets with GCB increased phosphorus digestibility (P ≤ 0.01) and suppressed ileum growth during the experimental period.

4. Results of the current study showed that phosphorus from gelatin can greatly improve broiler bone characteristics and phosphorus digestibility and complete elimination of inorganic phosphate sources from broiler diets is feasible with inclusion of 36 g/kg high phosphorus gelatin.     

miR-193 regulates proliferation of rat marrow mesenchymal stem cells by cell cycle-related proteins

AIM: To investigate the effects of microRNA-193 (miR-193) on the proliferation and apoptosis of rat bone marrow mesenchymal stem cells (MSCs). METHODS: Cultured rat MSCs were transfected with pre-miR-193 or anti-miR-193 to regulate the expression of miR-193. The proliferation of the MSCs after transfection was evaluated by MTS assay, colorimetric BrdU cell proliferation assay and Ki-67 immunostaining. Cell apoptosis was analyzed by flow cytometry with Annexin V/PI staining. The effect of miR-193 on the expression of cell cycle-related proteins was evaluated by qRT-PCR. RESULTS: Transfection of pre-miR-193 or anti-miR-193 regulated the expression of miR-193 in MSCs effectively. Over-expression of miR-193 significantly promoted the proliferation of MSCs (P<0.05), and inhibition of miR-193 reduced the proliferation of MSCs (P<0.05). miR-193 had no significant effect on the apoptosis of MSCs (P>0.05). The result of qRT-PCR indicated miR-193 promoted the expression of cyclin-dependent kinase 2 (CDK2) significantly (P<0.01). CONCLUSION: miR-193 promotes the proliferation of MSCs possibly through the CDK2 pathway.     

Differentiation of embryonic-like stem cells derived from human bone marrow into multinucleated fibers in vitro

AIM: To compare the capacity of in vitro differentiation into multinucleated fibers between embryonic-like stem cells (ELSCs) and mesenchymal stem cells (MSCs) derived from human bone marrow. METHODS: To isolate ELSCs, human bone marrow mononuclear cells were cultured in gelatin-coated flask with serum-free Knockout-DMEM medium designed for the expansion of human embryonic stem cells. MSCs were isolated from the same bone marrow by the traditional method. The morphological characters of both ELSCs and MSCs were observed under inverted phase-contrast microscope, and the expression of their multipotent antigen markers was identified by immunofluorescent staining. ELSCs and MSCs were cultured in myogenic differentiation medium. The protein levels of muscle-specific antigen markers myosin heavy chain (MHC), myogenin and MyoD were detected by the method of immunostaining. The mRNA expression of MHC, myogenin and MyoD was detected by RT-PCR. The capacity of in vitro differentiation into multinucleated fibers was compared between ELSCs and MSCs by calculating the proportion of MHC-positive multinucleated fibers. RESULTS: ELSCs, which weakly expressed the multipotential markers Oct-4, Nanog-3 and Sox-2, were isolated from bone marrow by the method of serum-free medium. ELSCs appeared smaller, slenderer and more homogeneous, and were morphologically different from MSCs derived from the same marrow. No multipotential marker in MSCs was expressed. ELSCs and MSCs were induced into long multinucleated fibers expressing MHC and myogenin at mRNA and protein levels by culturing in the myogenic differentiation medium. However, on the 10th day after induction, the proportion of the MHC-positive fibers in ELSCs was (25.7?4.1)%, and the proportion in MSCs was (15.8?7.6)%.The capacity for differentiation into muscle in ELSCs was significantly higher than that in MSCs (P<0.05). CONCLUSION: Bone marrow ELSCs are induced into multinucleated fibers and have the stronger myogenic differentiation capacity than MSCs derived from the same marrow. ELSCs are a more ideal candidate for muscular disease therapy.     

Adult human mesenchymal stem cell differentiates into adipocytes

AIM:To investigate the differentiation from human mesenchymal stem cells (hMSC) to adipocytes.METHODS: hMSC were separated from rib marrow and expanded in culture medium. To detect the surface antigens, the labeled cells were analysed on a FACScan flow cytometer. hMSC were induced with dexamethasone, insulin, 1-methy1-3-isobutylxanthine and indomethacin which acted as adipocyte differentiation inducer. The cells were stained with Oil Red O. The number of adipocytes were counted on a phase-contrast microscope.RESULTS: hMSC were expanded as undifferentiated cells in culture for more than 5 passages. The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. These expanded, attached MSC were uniformly positive for CD29, CD44, CD90, CD105, CD166 and didn't express CD14, CD34, CD45, CD11a. After induced with induction medium, lipid vacuoles were first detectable within the cells at 48 hours. Two weeks later, more than 85% MSC differentiated into adipocytes which displayed a perinuclear accumlation of lipid vacuoles, as detected by Oil Red O. CONCLUSION:hMSC can be induced to differentiate into adipocytes.     

PDL1Ig gene-modified BMSCs induce immune tolerance in rat liver transplantation

AIM: To investigate the effects of bone marrow mesenchymal stem cells(BMSCs) modified by programed death ligand-1 immunoglobulin(PDL₁Ig) gene on immune rejection of orthotopic liver transplantation in rats. METHODS: Rat BMSCs were cultured and identified. The protein expression of PDL₁Ig in the BMSCs 72 h after infection with pAdEasy-1/PDL₁Ig was detected by Western blot. Mixed lymphocyte reaction was used to detect the inhibitory effect of BMSCs on the viability of T-lymphocytes in peripheral blood. The male Wistar rats were used as donors(n=40), and the male SD rats were used as recipients(n=40). The rat model of orthotopic liver transplantation was established by improved cuff method for observing acute rejection. The rats were randomly divided into control group, BMSCs treatment group, BMSCs/GFP treatment group and BMSCs/PDL₁Ig treatment group with 10 pairs each. Five rats were executed at the 7th day and the remains were used for measuring the survival time. RESULTS: The expression of PDL₁Ig in the BMSCs was detected after pAdEasy-1/PDL₁Ig infection. The effect of BMSCs/PDL₁Ig on the viability of the lymphocytes was stronger than that of BMSCs/GFP. The level of IL-4 in BMSCs/PDL₁Ig group was significantly higher than that in the other 3 groups, while the levels of IFN-γ and IL-2 were significantly decreased. The liver function in BMSCs/PDL₁Ig group was significantly improved and the levels of ALT, AST and TBil were almost recovered to normal at the 7th day after transplantation. Severe rejection reaction was observed in control group, and rejection reactions were decreased with different degrees in BMSCs treatment group and BMSCs/GFP treatment group. Much slighter rejection reaction and significantly longer survival time were showed in BMSCs/PDL₁Ig group than those in the other 3 groups. CONCLUSION: PDL₁Ig-modified BMSCs inhibit the rejection of liver transplantation in rats and induce the immune tolerance, and the effect is better than that of BMSCs alone.