Suppression of MyoD induces spontaneous adipogenesis in skeletal muscle progenitor cell culture

The degree of intramuscular adipose tissue accumulation is one of the factors affecting meat quality. Accumulation of adipocytes is also observed under the pathological condition of skeletal muscle such as muscular dystrophy and sarcopenia. The origin of adipocytes seen in skeletal muscle is mesenchymal progenitor cells that can give rise to both adipocytes and fibroblasts. In the present study, we demonstrated that siRNA-mediated suppression of MyoD expression in rat skeletal muscle progenitor cell culture, which comprises both myogenic satellite cells and mesenchymal progenitor cells, resulted in diminished myotube formation and an unexpected spontaneous appearance of white adipocytes. Suppressing myomaker expression also resulted in complete absence of myotube formation without reducing MyoD expression, but no adipogenesis was seen in this scenario, indicating that decline in MyoD expression rather than decreased myotube formation is necessary to induce adipogenesis. In addition, spontaneous adipogenesis induced by suppressing MyoD expression in culture was inhibited by the conditioned medium from control culture, indicating that anti-adipogenic factor(s) are secreted from MyoD-positive myogenic cells. These results indicate the presence of regulatory mechanism on adipogenesis by myogenic cells.     

Genetic analysis for sow stayability at different parities in purebred Landrace and Large White pigs

Genetic parameters for sow stayability were estimated from farrowing records of 10,295 Landrace sows and 8192 Large White sows. The record for sow stayability from parity k to parity k + 1 (k = 1, …, 6) was 0 when a sow had a farrowing record at parity k but not at parity k + 1, and 1 when a sow had both records. Heritability was estimated by using single-trait linear and threshold animal models. Genetic correlations among parities were estimated by using two-trait linear–linear and single-trait random regression linear animal models. Genetic correlations with litter traits at birth were estimated by using a two-trait linear–linear animal model. Heritability estimates by linear model analysis were low (0.065–0.119 in Landrace & 0.061–0.157 in Large White); those by threshold model analysis were higher (0.136–0.200 & 0.110–0.283). Genetic correlations among parities differed between breeds and models. Genetic correlation between sow stayability and number born alive was positive in many cases, implying that selection for number born alive does not reduce sow stayability. The results seem to be affected by decisions on culling made by farmers.     

Identification of bovine serum albumin as an IgE-reactive beef component in a dog with food hypersensitivity against beef

IgE-reactive beef components were examined by an immunoblot analysis using a serum from a dog with food hypersensitivity against beef. The immunoblot analysis revealed a distinct band at approximately 66 kDa and a faint band at approximately 50 kDa. The immunoblot analysis for serum IgE reactivity to bovine serum albumin (BSA) also revealed a positive band at 66 kDa. Serum IgE reactivity to the 66-kDa protein of beef was diminished by pre-incubating the serum sample with BSA. Furthermore, a positive reaction to BSA was detected in intradermal testing in the dog. These results clearly indicated that BSA was an IgE-reactive beef component in the dog with food hypersensitivity against beef.     

Phylogenetic relationships of rodent pinworms (genus Syphacia) in Japan inferred from mitochondrial CO1 gene sequences

Species of the genus Syphacia are considered to have generally co-evolved with their rodent hosts. This study determined partial sequences of the CO1 gene from several species in the genus Syphacia and discuss the relationships between pinworms and their hosts. Syphacia montana, which parasitizes Microtinae, was closely related to S. frederici and S. obvelata, which parasitize Murinae. Although both S. obvelata and S. ohtaorum parasitize rodents in the genus Mus, these two species were not found to be closely related to each other. Syphacia frederici, S. emileromani and S. agraria are all pinworms of the Apodemus species, but genetic affiliation between these three species was not indicated. These facts suggest that the co-evolutionary relationship between species of the genus Syphacia and their host rodents may not so strict and host switching has probably occurred during the course of evolution.     

A novel culture method of canine peripheral blood lymphocytes with concanavalin a and recombinant human interleukin-2 for adoptive immunotherapy

This study was designed to develop a novel culture method for the efficient proliferation of canine peripheral blood lymphocytes (cPBL) for adoptive immunotherapy. When cPBL were cultured in the presence of concanavalin A (Con A), proliferation of cPBL was induced and expression of interleukin-2 receptor (IL-2R) which enables to respond to exogenously added IL-2 was upregulated. And then, when cPBL were cultured with recombinant human interleukin-2 (rhIL-2) in addition to Con A, proliferation was accelerated and increased to about 10-fold after 1 week. The phenotypic analysis showed that the main population of the cultured cPBL was consisted of CD8+ positive lymphocytes. Among them, CD4+CD8+ double positive (DP) lymphocytes had significantly increased, and the ratio of CD4+ single positive (SP) lymphocytes to CD8+ SP lymphocytes (CD4+SP/CD8+SP) was decreased as compared to before culturing. To evaluate the cytotoxic activity of cPBL cultured with Con A and rhIL-2, furthermore, cytotoxic assay was carried out against xenogeneic melanoma cell line (MeWo), which resulted in MHC-unrestricted cytokilling. These results suggest that the culture method of cPBL by the use of Con A and rhIL-2 may be useful for generating lymphokine activated killer cells, and also this may be beneficial for adoptive immunotherapy of tumor-bearing dogs.     

Vulnerability of feline T-lymphocytes to charged particles

An analysis of ionizing radiation-induced damage in peripheral lymphocytes has been employed to predict the prognosis of radiotherapy in terms of toxicity in normal tissues. Therefore, understanding the sensitivity of lymphocytes to high linear energy transfer (LET)-charged particles would be indispensable for utilizing charged particle therapy in veterinary medicine. However, the availability of such information is very limited. This study aimed to compare the radiosensitivity of feline T lymphocytes to gamma-rays (0.2 keV/microm) and 4 different types of charged particles with LET values ranging from 2.8 to 114 keV/microm. It was observed that the relative biological effectiveness, inactivation cross-section, and isodose-induced apoptosis increased in an LET-dependent manner. On the other hand, no difference in apoptosis frequency was observed in the cells exposed to an isosurvival dose of all the radiation types tested. This is the first study that demonstrates the LET dependence of cell killing and apoptosis induction in feline T lymphocytes. Our results suggest that lymphocytes can be effectively used to predict the prognosis of charged-particle therapy in cat patients.     

Efficient plant regeneration via organogenesis in “Egusi” melon (Colocynthis citrullus L.)

Plant regeneration protocol of “Egusi” melon (Colocynthis citrullus L.) was established using three local (“Ejagham”, “Sewere” and “Barablackedge”) and one improved (NHC1-130) cultivars. Cotyledonary explants of different lengths (1/2, 1/4 and 1/6) excised from 4- or 8-day-old seedlings germinated in vitro were cultured on MS medium supplemented with different concentrations of 6-benzylaminopurine (BA). The best results were obtained when cotyledons from 4-day-old seedlings were cut into 2 (1/2) halves. Plant regeneration was optimal on medium containing 5 mg/l BA, yielding 86.3%, 77.0% and 76.3% shoot induction frequencies amongst the three local cultivars of “Ejagham”, “Sewere” and “Barablackedge”, respectively. In NHC1-130, the highest shoot induction frequency (85%) was obtained on medium containing 2 mg/l BA. Adventitious shoots were elongated on medium containing 0.1 mg/l BA and successfully rooted on hormone-free MS medium. Flow cytometric analysis revealed 70% of the plants to be diploid.     

Effects of instream structures on Kirikuchi charr, southernmost population of genus Salvelinus, in a small mountain stream of Japan

ABSTRACT:   The changes in physical habitats and responses of fish, e.g. the Kirikuchi charr Salvelinus leucomaenis japonicus , were examined by placing instream structures in a stream on the upper drainage of the Totsu River system of the Kii Peninsula, Japan, over a two-year period. Instream structures created pools below the structures, and these pools have maintained their functions as fish habitats over two years. In two treatment sections, sections A and B (some red-spotted masu salmon were removed in advance in section B), mean water depth increased soon after placing the structures, but decreased over two years. Mean water velocity slowed after placing the structures in both sections. Total fish abundance has been relatively high at the two treatment sections compared with a control section after placing the structures. Age 1 Kirikuchi charr, however, increased only in section B. These results indicate that instream structures can serve as habitat enhancement for fish, including Kirikuchi charr, in the study area if other appropriate habitat factors for respective species are taken into consideration.