Choi Sj

Long-term cultivation of multipotential neural stem cells from adult rat subependyma.

Cultivation of adult rat neural stem cells (RNSCs) from the ventricular subependyma has been reported to be more difficult than growth of mouse neural stem cells. This is unfortunate, because rats provide useful models of brain function and disease, and implantation of RNSCs in these models could provide critical information on allograft behavior. Growing the cells in an appropriate medium (NS-A+B27 supplement), plating at sufficient densities (>5 cells per mm(2)), and minimizing opportunities for detachment from the substratum made it possible to isolate and cultivate these cells for over 6 months for >50 passages with no apparent change in phenotype. Single clones could be expanded indefinitely and differentiated to form astrocytes, oligodendrocytes, and neurons, demonstrating that the cultures did indeed contain neural stem cells. The cells had a much shorter cell cycle time ( approximately 13 h) than doubling time ( approximately 35 h), suggesting that these cells produce post-mitotic cells in approximately two of three divisions, thus making expansion difficult. The optimization of methods to grow adult RNSCs and identification of characteristics that limit their growth should prove useful in increasing the use of RNSCs for studies of their potential role in brain health and disease.

Paradoxical effects of elastase inhibitor guamerin on the tissue repair of two different wound models: sealed cutaneous and exposed tongue wounds

Innate elastase inhibitors are known to be putatively involved in the regulation of tissue inflammation by inhibiting polymorphonuclear leukocyte (PMN) derived proteinases. The aim of this study was to evaluate affects of leukocyte elastase suppression and PMN infiltration on wound healing in mouse by administering the recombinant elastase inhibitor guamerin (rEIG) in two different wound models; 1) impaired pin-punctured dorsal mucosa of anterior tongue wound, 60 mice, treated with saline containing rEIG that were fed ad libitum and 2) stable linear excisional cutaneous wound, 40 mice, covered with fibrin sealant containing rEIG. The progress of healing was analyzed by histological methods. The tongue wounds treated with rEIG became edematous around the pin-punctured tongue wound, and influx of inflammatory cells and PMN into the underlying stromal tissue were seen rapidly after wounding and peaked between 2-4 days. Whereas the control mice showed almost no wheal formation in the pin-punctured wound, a far lesser levels of PMN infiltration, and almost complete wound closure in 4 days. In the other model, the liner excisional cutaneous wound treated with fibrin sealant containing rEIG showed early wound constriction, lesser degree of inflammatory cells influx, and complete reepithelialization in 4-5 days, whereas the wound of control mice with the fibrin sealant alone showed contrary delayed reepithelialization, greater degree of inflammatory cell infiltration, and consequencial formation of greater granulation tissue at wound site. Taken together, these data suggest paradoxical effects of rEIG on the wound healing where in the wound exposed to infiltrating milieu of microorganisms in the oral cavity, the rEIG aggravates the wound healing by interfering with other innate defensive factors and extended greater flux of PMNs to inflamed wound site, while in the wound enclosed by fibrin, the rEIG accelerated wound healing by inhibiting the inflammation-generated proteases and the acute inflammatory reaction.

Posteroventral pallidotomy in medically intractable postapoplectic monochorea: case report

BACKGROUND Posteroventral pallidotomy is a widely accepted surgical procedure for treating medically intractable Parkinsons disease and Levo-dopa induced dyskinesia. In the surgical treatment of hyperkinetic movement disorders, generalized dystonia has recently become a favorable indication of posteroventral pallidotomy. However, a commonly recognized surgical procedure for treating choreiform movement disorders has not yet been established. Here we present an unusual experience of a posteroventral pallidotomy performed to treat a medically intractable monochorea caused by a vascular insult on the basal ganglia. METHODS A 63-year-old female presented with choreiform movement of the left upper limb that she had suffered for 5 months. She was found to have a hemorrhagic infarction in the right putaminal area. No other abnormal lesions were shown by magnetic resonance imaging except for a widening of the right cerebellopontine cistern because of an acoustic neurinoma removed 5 years previously. Despite medication with a dopamine antagonist, choreiform movement of the left limb had not improved, and the patient complained of rigidity and slowness of ambulation owing to the side effects of the medicine. A right posteroventral pallidotomy was performed with macrostimulation for a physiologic confirmation of the globus pallidus internus (GPi), which is the conventional target for Parkinsons disease. After coagulating the GPi target, the choreiform movement of the contralateral upper limb was completely abolished. RESULTS The postoperative course was uneventful and no recurrence of chorea was observed over a follow-up period of 6 months. CONCLUSIONS Stereotactic surgery for hyperkinetic movement disorders is not as common a procedure as that used for treating Parkinsons disease. Furthermore, there have been few reports of pallidal surgery for treating the chorea caused by an ischemic insult. However, on the basis of the current concept that varying types of hyperkinetic disorders may have a common pathophysiological mechanism, a posteroventral pallidotomy may be an alternative surgical procedure for treating medically intractable postapoplectic chorea like in an occasion of dystonia.

Kruppel-like factor 4 positively regulates autoimmune arthritis in mouse models and rheumatoid arthritis in patients via modulating cell survival and inflammation factors of fibroblast-like synoviocyte

Rheumatoid arthritis (RA) is a chronic autoimmune disease that causes mild to severe joint inflammation. During RA pathogenesis, fibroblast-like synoviocytes (FLS) acquire a tumor-like phenotype and mediate cartilage destruction both directly and indirectly by producing proinflammatory cytokines and matrix metalloproteinases (MMPs). Kruppel-like factor (KLF) 4, a member of the KLF family, plays significant roles in cell survival, proliferation, and differentiation. A recent study reported increased expression of KLF4 in synovial tissue from RA patients. However, its precise role in RA in different models, including mouse autoimmune disease models, remains unclear. In this study, we examined the role of KLF4 during development of autoimmune arthritis in mouse models. To do this, we used KLF4 knockout mice rendered by ribonucleic acid (RNA)-guided endonuclease (RGEN) and performed collagen antibody-induced arthritis (CAIA). We found that deletion of KLF4 reduces inflammation induced by CAIA. In addition, we assessed collagen-induced arthritis (CIA) in control mice and KLF4-overexpressing mice generated by a minicircle vector treatment. Severity of CIA in mice overexpressing KLF4 was greater than that in mice injected with control vector. Finally, we verified the inflammatory roles of KLF4 in CIA by treating Kenpaullone which is used as KLF4 inhibitor. Next, we focused on human/mouse FLS to discover the cellular process involved in RA pathogenesis including proliferation, apoptosis, and inflammation including MMPs. In FLS, KLF4 upregulated expression of mRNA encoding proinflammatory cytokines interleukin (IL)-1β and IL-6. KLF4 also regulated expression of matrix metallopeptidase 13 in the synovium. We found that blockade of KLF4 in FLS increased apoptosis and suppressed proliferation followed by downregulation of antiapoptotic factor BCL2. Our results indicate that KLF4 plays a crucial role in pathogenesis of inflammatory arthritis in vivo, by regulating apoptosis, MMP expression, and cytokine expression by FLS. Thus, KLF4 might be a novel transcription factor for generating RA by modulating cellular process of FLS.

Interrupting oral infection of Porphyromonas gingivalis with anti-FimA antibody attenuates bacterial dissemination to the arthritic joint and improves experimental arthritis

Rheumatoid arthritis (RA) is a chronic autoimmune disease that typically results in strong inflammation and bone destruction in the joints. It is generally known that the pathogenesis of RA is linked to cardiovascular and periodontal diseases. Though rheumatoid arthritis and periodontitis share many pathologic features such as a perpetual inflammation and bone destruction, the precise mechanism underlying a link between these two diseases has not been fully elucidated. Collagen-induced arthritis (CIA) mice were orally infected with Porphyromonas gingivalis (Pg) or Pg preincubated with an anti-FimA antibody (FimA Ab) specific for fimbriae that are flexible appendages on the cell surface. Pg-infected CIA mice showed oral microbiota disruption and increased alveolar bone loss and had synovitis and joint bone destruction. However, preincubation with FimA Ab led to a significant reduction in the severity of both oral disease and arthritis. Moreover, FimA Ab attenuated bacterial attachment and aggregation on human gingival and rheumatoid arthritis synovial fibroblasts. In addition, we discovered bacteria may utilize dendritic cells, macrophages and neutrophils to migrate into the joints of CIA mice. These results suggest that disrupting Pg fimbriae function by FimA Ab ameliorates RA.