Complete remission of diabetes with a transient HDAC inhibitor and insulin in streptozotocin mice

Despite the growing epidemic worldwide, diabetes is an incurable disease. We have been focusing on why diabetes manifests refractoriness to any therapy. We recently found that abnormal bone marrow-derived cells (BMDCs), namely, Vcam-1+ST-HSCs, was a key mechanism for diabetic complications. We then hypothesize that those aberrant BMDCs sustainedly impair pancreatic β cells. Here we show that eliminating abnormal BMDCs using bone marrow transplantation results in controlling serum glucose in diabetic mice, in which normoglycemia is sustained even after cessation of insulin therapy. Alternatively, abnormal BMDCs exhibiting epigenetic alterations are treated with an HDAC inhibitor, givinostat, in diabetic mice. As a result, those mice are normoglycemic along with restored insulin secretion even following the cessation of both insulin and givinostat. Diabetic cell fusion between abnormal BMDCs and resident cells is significantly blocked by the combination therapy in the pancreatic islets and thymus while surgical ablation of the thymus completely eliminates therapeutic protection in diabetic mice. In conclusion, diabetes is an epigenetic stem cell disorder with thymic disturbances. The combination may be applied to patients aiming at complete remission from diabetes in clinical medicine.journal articl

Spatiotemporal expression of TRPM4 in the mouse cochlea

The present study was conducted to elucidate the presence of the transient receptor potential cation channel subfamily M member 4, TRPM4, in the mouse inner ear. TRPM4 immunoreactivity (IR) was found in the cell body of inner hair cells (IHCs) in the organ of Corti in the apical side of marginal cells of the stria vascularis, in the apical portion of the dark cells of the vestibule, and in a subset of the type II neurons in the spiral ganglion. Subsequently, changes in the distribution and expression of TRPM4 in the inner ear during embryonic and postnatal developments were also evaluated. Immunohistochemical localization demonstrated that the emergence of the TRPM4-IR in IHCs occurs shortly before the onset of hearing, whereas that in the marginal cells happens earlier, at the time of birth, coinciding with the onset of endolymph formation. Furthermore, semiquantitative real-time PCR assay showed that expressions of TRPM4 in the organ of Corti and in the stria vascularis increased dramatically at the onset of hearing. Because TRPM4 is a Ca2+-activated monovalent-selective cation channel, these findings imply that TRPM4 contributes to potassium ion transport, essential for the signal transduction in IHCs and the formation of endolymph by marginal cells. © 2014 Wiley Periodicals, Inc

Strategies of community health activities to improve the quality of inhabitants' life

ヘルスプロモーションの理念にそって，住民組織，行政，研究グループが協働して，住民の生活の質の向上をめざした地域保健活動の展開を計画した。まず，休眠状態にあった岡山県M町の健康づくり推進協議会（以下M町協議会と略す）を活性化するため，協議会の委員18人を対象に聴き取り調査を行い，次に，M町協議会でのディスカッションを深めた。1．M町協議会構成団体のうち住民組織は9組織で，このうち6組織は居住地域内の住民の推薦や輪番で代表を選出しており，他の3組織は任意であった。8組織の事務局は行政機関内にあり，活動経費の公費補助率は70％以上であった。2．委員の描く理想の町のイメージとして，福祉の充実した町で老後も安心できるという内容を含んだ回答が多かった。それを実現する為に，住民一人一人が自己管理意識を持つと同時に，協議会が支援的な活動を身近なところで展開することの必要性が挙げられた。3．M町協議会活動への委員の期待は大きく，協議会内での委員間の意思疎通も次第に深められた。事前に聴き取り調査を行ったこと等が効果的に影響したと考えられた。Along with the idea of health promotion, we planned the community health activities to improve the quality of inhabitants' life in cooperation with inhabitants, community organizations, local government and health professionals. In order to revitalize an inactive committee for health promotion of M town hearing surveys were conducted on the members of the committee. 1. Leaders of community organizations occupied 9 of 19 members of the committee. Leaders of 6 organizations out of 9 ones were decided by recommendation or rotation. Eight secretariats of those organizations were located in the governmental office. Seventy percent or more of activity costs of these organizations depends on assistance of local government. 2. Members of the committee imaged the town, which was healthy and filled with mind of welfare as an ideal town. They pointed out that it was required for the committee to perform support activities to realize an ideal town as well as for every inhabitant to have the consciousness of self-management. 3. Activities of the committee were much expected by members of the committee. Communications between members deepened gradually by repeating the meetings. Hearing survey on members also seemed to be much effective for those progress

A Case of Super-giant Basal Cell Carcinoma Initially Diagnosed as Multiple Traumas

Basal cell carcinoma (BCC), which is relatively easy to diagnose in a clinical setting, is the most common malignant tumor in the skin. Conversely, a giant BCC, a tumor beyond 5 cm in diameter, is a rare disease. In particular, a giant BCC beyond 20 cm in diameter is called a super-giant BCC, which frequently invades into deeper tissues, including the dermis, bones, or muscles. Here, we present a case of a 71-year-old patient who was initially diagnosed with multiple traumas with a large periosteal defect of the head. The ulcer was surrounded by malodorous necrotic tissue and slough, and several bacteria that caused necrotizing fasciitis were detected. Mapping biopsies after extensive debridement yielded BCC, and therefore, he was finally diagnosed with a super-giant BCC. A careful consultation revealed a history of ulcer on the head after a head injury approximately 10 years ago. He underwent radical dissection including the external table of the skull, followed by a free latissimus dorsi muscle flap with a meshed split-thickness skin graft. Because of the slow and chronic development of a super-giant BCC, accurate diagnosis is often difficult. Careful attention should be paid in patients with long-sustained ulcers

Gene Therapy for Neuropathic Pain through siRNA-IRF5 Gene Delivery with Homing Peptides to Microglia.

Astrocyte- and microglia-targeting peptides were identified and isolated using phage display technology. A series of procedures, including three cycles of both in vivo and in vitro biopanning, was performed separately in astrocytes and in M1 or M2 microglia,yielding 50-58 phage plaques in each cell type. Analyses of the sequences of this collection identified one candidate homing peptide targeting astrocytes (AS1[C-LNSSQPS-C]) and two candidate homing peptides targeting microglia (MG1[C-HHSSSAR-C] and MG2[C-NTGSPYE-C]). To determine peptide specificity for the target cell in vitro, each peptide was synthesized and introduced into the primary cultures of astrocytes or microglia. Those peptides could bind to the target cells and be selectively taken up by the corresponding cell, namely, astrocytes, M1 microglia, or M2 microglia. To confirm cell-specific gene delivery to M1 microglia, the complexes between peptide MG1 and siRNA-interferon regulatory factor 5 were prepared and intrathecally injected into a mouse model of neuropathic pain. The complexes successfully suppressed hyperalgesia with high efficiency in this neuropathic pain model. Here, we describe a novel gene therapy for the treatment neuropathic pain, which has a high potential to be of clinical relevance. This strategy will ensure the targeted delivery of therapeutic genes while minimizing side effects to non-target tissues or cells

GLT1 gene delivery based on bone marrow-derived cells ameliorates motor function and survival in a mouse model of ALS.

Amyotrophic lateral sclerosis (ALS) is an intractable neurodegenerative disease. CD68-positive bone marrow (BM)-derived cells (BMDCs) accumulate in the pathological lesion in the SOD1(G93A) ALS mouse model after BM transplantation (BMT). Therefore, we investigated whether BMDCs can be applied as gene carriers for cell-based gene therapy by employing the accumulation of BMDCs. In ALS mice, YFP reporter signals were observed in 12-14% of white blood cells (WBCs) and in the spinal cord via transplantation of BM after lentiviral vector (LV) infection. After confirmation of gene transduction by LV with the CD68 promoter in 4-7% of WBCs and in the spinal cord of ALS mice, BM cells were infected with LVs expressing glutamate transporter (GLT) 1 that protects neurons from glutamate toxicity, driven by the CD68 promoter, which were transplanted into ALS mice. The treated mice showed improvement of motor behaviors and prolonged survival. Additionally, interleukin (IL)-1β was significantly suppressed, and IL-4, arginase 1, and FIZZ were significantly increased in the mice. These results suggested that GLT1 expression by BMDCs improved the spinal cord environment. Therefore, our gene therapy strategy may be applied to treat neurodegenerative diseases such as ALS in which BMDCs accumulate in the pathological lesion by BMT

Hyperglycemia Induces Skin Barrier Dysfunctions with Impairment of Epidermal Integrity in Non-Wounded Skin of Type 1 Diabetic Mice.

Diabetes causes skin complications, including xerosis and foot ulcers. Ulcers complicated by infections exacerbate skin conditions, and in severe cases, limb/toe amputations are required to prevent the development of sepsis. Here, we hypothesize that hyperglycemia induces skin barrier dysfunction with alterations of epidermal integrity. The effects of hyperglycemia on the epidermis were examined in streptozotocin-induced diabetic mice with/without insulin therapy. The results showed that dye leakages were prominent, and transepidermal water loss after tape stripping was exacerbated in diabetic mice. These data indicate that hyperglycemia impaired skin barrier functions. Additionally, the distribution of the protein associated with the tight junction structure, tight junction protein-1 (ZO-1), was characterized by diffuse and significantly wider expression in the diabetic mice compared to that in the control mice. In turn, epidermal cell number was significantly reduced and basal cells were irregularly aligned with ultrastructural alterations in diabetic mice. In contrast, the number of corneocytes, namely, denucleated and terminally differentiated keratinocytes significantly increased, while their sensitivity to mechanical stress was enhanced in the diabetic mice. We found that cell proliferation was significantly decreased, while apoptotic cells were comparable in the skin of diabetic mice, compared to those in the control mice. In the epidermis, Keratin 5 and keratin 14 expressions were reduced, while keratin 10 and loricrin were ectopically induced in diabetic mice. These data suggest that hyperglycemia altered keratinocyte proliferation/differentiation. Finally, these phenotypes observed in diabetic mice were mitigated by insulin treatment. Reduction in basal cell number and perturbation of the proliferation/differentiation process could be the underlying mechanisms for impaired skin barrier functions in diabetic mice

Aberrant bone marrow-derived microglia in the hypothalamus may dysregulate appetite in diabetes

Bone marrow derived cells (BMDCs) migrate into the hypothalamus, where those cells give rise to microglia to regulate food intake. Given the fact that diabetes functionally impairs BMDCs, we hypothesized that diabetic microglia would fail to exhibit physiological function, accounting for hyperphagia in diabetes. To examine the role of BMDCs, total bone marrow cells from GFP transgenic mice were transplanted into wild type mice in which diabetes was induced by streptozotocin. We first confirmed that bone marrow transplantation could be utilized to examine BMDCs in the brain parenchyma as GFP positive cells could engraft the brain parenchyma and give rise to microglia even when the BBB was intact in the recipient mice. While diabetic mice manifested hyperphagia, BMDCs were in smaller number in the hypothalamus with less response to fasting in the brain parenchyma compared to nondiabetic mice. This finding was also confirmed by examining nondiabetic chimera mice in which BMDCs were diabetic. Those mice also exhibited less response of BMDCs in response to fasting. In conclusion, diabetic BMDCs had less response of microglia to fasting, perhaps accounting for diabetic hyperphagia.journal articl

Superchargeを付加した腹直筋皮弁を用いた乳房再建

2013年以降、乳がん全摘後に保険適応となった人工ゲル充填乳房を用いた乳房再建術のまれな合併症として、T細胞型非ホジキンリンパ腫が報告され、同タイプの人工ゲル充填乳房は使用できなくなった。これを受けて、当院では自家組織を用いた乳房再建の症例が増加している。ボリュームのある乳房の場合、腹部を用いた有茎腹直筋皮弁、遊離皮弁が選択されることが多い。有茎皮弁では上腹壁動静脈を血管茎とするが、腹直筋の血行は深下腹壁動静脈優位のため血行が不十分と考えられる硬結が術後に生じることがある。これを改善するため、当科では可能な限り深下腹壁動静脈を胸部のレシピエント血管に吻合してsuperchargeを行い、血行の増強に努めている。遊離皮弁と異なり、皮弁の自由度が制限されるため血管吻合がやや困難であるが、術後は柔らかい乳房が再現されることが期待できる。With the onset of insurance coverage in 2013, implant-based techniques had been popular for breast reconstruction in Japan until fairly recently. However, possible complications of non-Hodgkin’s lymphoma caused by implant materials led a recall of textured-typed implants for breast reconstruction. Such situation led patients head for breast reconstruction using autologous tissue. In case of well-developed breasts, either rectus abdominal musculocutaneous flap or deep inferior epigastric perforator flap is frequently selected for breast reconstruction. The former flap is well-vascularized but rectus abdominal muscle is used to transpose the flap, which could cause the herniation of the abdominal wall as a complication at late-stage. In the latter flap, since abdominal fat tissue is used, reconstructed breasts are similar to original breasts, while operation periods are relatively long in order to anastomose vessels under microscopy. Besides, thrombosis in anastomosed vessels could cause the total necrosis of reconstructed breasts.To overcome the disadvantages of the both methods, rectus abdominal musculocutaneous flaps anastomosed with vessels in the recipient region (i.e., Supercharged flap) have been developed. Here, we will describe seven cases with supercharged flaps to reconstruct breasts performed in our hospital