Generation of CD2+CD8+ NK Cells from c-kit+ Bone Marrow Cells in Porcine

Natural killer (NK) cells provide one of the initial barriers of cellular host defense against pathogens, in particular intracellular pathogens. Because bone marrow-derived hematopoietic stem cells (HSCs), lymphoid protenitors, can give rise to NK cells, NK ontogeny has been considered to be exclusively lymphoid. Here, we show that porcine c-kit+ bone marrow cells (c-kit+ BM cells) develop into NK cells in vitro in the presence of various cytokines [interleukin (IL)-2, IL-7, IL-15, IL-21, stem cell factor (SCF), and fms-like tyrosine kinase-3 ligand (FLT3L)]. Adding hydrocortisone (HDC) and stromal cells greatly increases the frequency of c-kit+ BM cells that give rise to CD2+CD8+ NK cells. Also, intracellular levels of perforin, granzyme B, and NKG2D were determined by RT-PCR and western blotting analysis. It was found that of perforin, granzyme B, and NKG2D levels significantly were increased in cytokine-stimulated c-kit+ BM cells than those of controls. And, we compared the ability of the cytotoxicity of CD2+CD8+ NK cells differentiated by cytokines from c-kit+ BM cells against K562 target cells for 28 days. Cytokines-induced NK cells as effector cells were incubated with K562 cells as target in a ratio of 100:1 for 4 h once a week. In results, CD2+CD8+ NK cells induced by cytokines and stromal cells showed a significantly increased cytotoxicity 21 days later. Whereas, our results indicated that c-kit+ BM cells not pretreated with cytokines have lower levels of cytotoxicity. Taken together, this study suggests that cytokines-induced NK cells from porcine c-kit+ BM cells may be used as adoptive transfer therapy if the known obstacles to xenografting (e.g. immune and non-immune problems) were overcome in the future.

Pain modality and spinal glia expression by streptozotocin induced diabetic peripheral neuropathy in rats / 한국실험동물학회지

Pain symptoms are a common complication of diabetic peripheral neuropathy or an inflammatory condition. In the most experiments, only one or two evident pain modalities are observed at diabetic peripheral neuropathy according to experimental conditions. Following diabetic peripheral neuropathy or inflammation, spinal glial activation may be considered as an important mediator in the development of pain. For this reason, the present study was aimed to address the induction of pain modalities and spinal glial expression after streptozotocin injection as compared with that of zymosan inflammation in the rat. Evaluation of pain behavior by either thermal or mechanical stimuli was performed at 3 weeks or 5 hours after either intravenous streptozotocin or zymosan. Degrees of pain were divided into 4 groups: severe, moderate, mild, and non-pain induction. On the mechanical allodynia test, zymosan evoked predominantly a severe type of pain, whereas streptozotocin induced a weak degree of pain (severe+moderate: 57.1%). Although zymosan did not evoke cold allodynia, streptozotocin evoked stronger pain behavior, compared with zymosan (severe+moderate: 50.0%). On the other hand, the high incidence of thermal hyperalgesia (severe+moderate: 90.0%) and mechanical hyperalgesia (severe+moderate: 85.7%) by streptozotocin was observed, as similar to that of zymosan. In the spinal cord, the increase of microglia and astrocyte was evident by streptozotocin, only microglia was activated by zymosan. Therefore, it is recommended that the selection of mechanical and thermal hyperalgesia is suitable for the evaluation of streptozotocin induced diabetic peripheral neuropathy. Moreover, spinal glial activation may be considered an important factor.

The Antinociceptive Effect of Sigma-1 Receptor Antagonist, BD1047, in a Capsaicin Induced Headache Model in Rats

Intracranial headaches, including migraines, are mediated by nociceptive activation of the trigeminal nucleus caudalis (TNC), but the precise mechanisms are poorly understood. We previously demonstrated that selective blockage of spinal sigma-1 receptors (Sig-1R) produces a prominent antinociceptive effect in several types of pain models. This study evaluates whether the Sig-1R antagonist (BD1047) has an antinociceptive effect on capsaicin (a potent C-fiber activator) induced headache models in rats. Intracisternal infusion of capsaicin evoked pain behavior (face grooming), which was significantly attenuated by BD1047 pretreatment. BD1047 consistently reduced capsaicin-induced Fos-like immunoreactivity (Fos-LI), a neuronal activator, in the TNC in a dose-dependent manner. Moreover, capsaicin-induced phosphorylation of N-methyl-D-aspartate receptor subunit 1 was reversed by BD1047 pretreatment in the TNC. These results indicate that the Sig-1R antagonist has an inhibitory effect on nociceptive activation of the TNC in the capsaicin-induced headache animal model.

A Survey of Ectoparasite Infestations in Stray Dogs of Gwang-ju City, Republic of Korea

This study was designed to investigate the incidence of ectoparasite infestation among stray dogs in Gwang-ju City, Republic of Korea. A total of 103 stray dogs collected in the Animal Shelter of Gwang-ju City from November 2003 to August 2005 were investigated in this study. Ectoparasites of one or more genera were detected in 45.6% (47 / 103) of the dogs examined for dermatologic lesions and/or skin scrapings (from 3-5 affected areas). Otodectes cynotis was found to be the most frequent parasite (22.3%, 23 / 103), followed by Sarcoptes scabiei var canis (19.4%, 20 / 103), Ctenocephalides canis (6.8%, 7 / 103), Demodex canis (4.9%, 5 / 103), and Trichodectes canis (1.0%, 1 / 103). Monospecific infestation was found in 83.0% (39 / 47) of the affected dogs, whereas concurrent infestations with 2 or more ectoparasites per animal were found in 17.0% (8 / 47) of the affected dogs. Trichodectes canis is reported for the first time in the Republic of Korea. Dogs less than 1 yr old were more heavily infected than other age groups (66.7%), and small-sized dogs of less than 3 kg body weight were more heavily infected than larger dogs (41.7%).

A Survey of Ectoparasite Infestations in Stray Dogs of Gwang-ju City, Republic of Korea

This study was designed to investigate the incidence of ectoparasite infestation among stray dogs in Gwang-ju City, Republic of Korea. A total of 103 stray dogs collected in the Animal Shelter of Gwang-ju City from November 2003 to August 2005 were investigated in this study. Ectoparasites of one or more genera were detected in 45.6% (47 / 103) of the dogs examined for dermatologic lesions and/or skin scrapings (from 3-5 affected areas). Otodectes cynotis was found to be the most frequent parasite (22.3%, 23 / 103), followed by Sarcoptes scabiei var canis (19.4%, 20 / 103), Ctenocephalides canis (6.8%, 7 / 103), Demodex canis (4.9%, 5 / 103), and Trichodectes canis (1.0%, 1 / 103). Monospecific infestation was found in 83.0% (39 / 47) of the affected dogs, whereas concurrent infestations with 2 or more ectoparasites per animal were found in 17.0% (8 / 47) of the affected dogs. Trichodectes canis is reported for the first time in the Republic of Korea. Dogs less than 1 yr old were more heavily infected than other age groups (66.7%), and small-sized dogs of less than 3 kg body weight were more heavily infected than larger dogs (41.7%).