Novel Autologous Therapy for Long-Gap Peripheral Nerve Injury Using Human Sk-SCs

Losses in vital functions of the somatic motor and sensory nervous system are induced by severe long-gap peripheral nerve transection injury. In such cases, autologous nerve grafts are the gold standard treatment, despite the unavoidable sacrifice of other healthy functions, whereas the prognosis is not always favorable. Here, we use human skeletal muscle-derived stem cells (Sk-SCs) to reconstitute the function after long nerve-gap injury. Muscles samples were obtained from the amputated legs from 9 patients following unforeseen accidents. The Sk-SCs were isolated using conditioned collagenase solution, and sorted as CD34+/45- (Sk-34) and CD34-/45-/29+ (Sk-DN/29+) cells. Cells were separately cultured/expanded under optimal conditions for 2 weeks, then injected into the athymic nude mice sciatic nerve long-gap model (7-mm) bridging an acellular conduit. After 8-12 weeks, active cell engraftment was observed only in the Sk-34 cell transplanted group, showing preferential differentiation into Schwann cells and perineurial/endoneurial cells, as well as formation of the myelin sheath and perineurium/endoneurium surrounding regenerated axons, resulted in 87% of numerical recovery. Differentiation into vascular cell lineage (pericyte and endothelial cells) were also observed. A significant tetanic tension recovery (over 90%) of downstream muscles following electrical stimulation of the sciatic nerve (at upper portion of the gap) was also achieved. In contrast, Sk-DN/29+ cells were completely eliminated during the first 4 weeks, but relatively higher numerical (83% vs. 41% in axon) and functional (80% vs. 60% in tetanus) recovery than control were observed. Noteworthy, significant increase in the formation of vascular networks in the conduit during the early stage (first 2 weeks) of recovery was observed in both groups with the expression of key factors (mRNA and protein levels), suggesting the paracrine effects to angiogenesis. These results suggested that the human Sk-SCs may be a practical source for autologous stem cell therapy following severe peripheral nerve injury

Identification of myogenic-endothelial progenitor cells in the interstitial spaces of skeletal muscle

Putative myogenic and endothelial (myo-endothelial) cell progenitors were identified in the interstitial spaces of murine skeletal muscle by immunohistochemistry and immunoelectron microscopy using CD34 antigen. Enzymatically isolated cells were characterized by fluorescence-activated cell sorting on the basis of cell surface antigen expression, and were sorted as a CD34+ and CD45− fraction. Cells in this fraction were ∼94% positive for Sca-1, and mostly negative (<3% positive) for CD14, 31, 49, 144, c-kit, and FLK-1. The CD34+/45− cells formed colonies in clonal cell cultures and colony-forming units displayed the potential to differentiate into adipocytes, endothelial, and myogenic cells. The CD34+/45− cells fully differentiated into vascular endothelial cells and skeletal muscle fibers in vivo after transplantation. Immediately after sorting, CD34+/45− cells expressed only c-met mRNA, and did not express any other myogenic cell-related markers such as MyoD, myf-5, myf-6, myogenin, M-cadherin, Pax-3, and Pax-7. However, after 3 d of culture, these cells expressed mRNA for all myogenic markers. CD34+/45− cells were distinct from satellite cells, as they expressed Bcrp1/ABCG2 gene mRNA (Zhou et al., 2001). These findings suggest that myo-endothelial progenitors reside in the interstitial spaces of mammalian skeletal muscles, and that they can potentially contribute to postnatal skeletal muscle growth

Two cases of pulmonary dirofilariasis

Pulmonary dirofilariasis is almost always asymptomatic. An abnormal nodule was accidentally discovered by a chest x-ray during a medical checkup and detailed examination for other diseases was performed. Case １. A female patient, in her ７０s, was admitted owing to flu-like symptoms and bloody sputum. A chest computed tomography（CT）scan revealed a nodular shadow with １０‐mm ground glass opacity in the right lower lobe. Case２. A female patient, in her ６０s, was admitted due to an abnormal shadow on a chest xray in the left lung during a regular medical checkup. A chest CT scan revealed a nodular shadow with １３‐mm ground glass opacity in the left upper lobe. Lung cancer was suspected in both cases. Thoracoscopy and partial lung resection were performed to confirm the diagnosis. The specimen consisted of granulation tissue and no malignancy was found at operation. Final pathological diagnosis revealed pulmonary dirofilariasis. Pulmonary dirofilariasis can be definitively diagnosed by detecting a worm body. We believe partial lung resection during video-assisted thoracic surgery is a minimally invasive and an effective treatment for this disease

Cardiomyocyte Formation by Skeletal Muscle-Derived Multi-Myogenic Stem Cells after Transplantation into Infarcted Myocardium

BACKGROUND: Cellular cardiomyoplasty for myocardial infarction has been developed using various cell types. However, complete differentiation and/or trans-differentiation into cardiomyocytes have never occurred in these transplant studies, whereas functional contributions were reported. METHODS AND RESULTS: Skeletal muscle interstitium-derived CD34(+)/CD45(-) (Sk-34) cells were purified from green fluorescent protein transgenic mice by flowcytometory. Cardiac differentiation of Sk-34 cells was examined by in vitro clonal culture and co-culture with embryonic cardiomyocytes, and in vivo transplantation into a nude rat myocardial infarction (MI) model (left ventricle). Lower relative expression of cardiomyogenic transcription factors, such as GATA-4, Nkx2-5, Isl-1, Mef2 and Hand2, was seen in clonal cell culture. However, vigorous expression of these factors was seen on co-culture with embryonic cardiomyocytes, together with formation of gap-junctions and synchronous contraction following sphere-like colony formation. At 4 weeks after transplantation of freshly isolated Sk-34 cells, donor cells exhibited typical cardiomyocyte structure with formation of gap-junctions, as well as intercalated discs and desmosomes, between donor and recipient and/or donor and donor cells. Fluorescence in situ hybridization (FISH) analysis detecting the rat and mouse genomic DNA and immunoelectron microscopy using anti-GFP revealed donor-derived cells. Transplanted Sk-34 cells were incorporated into infarcted portions of recipient muscles and contributed to cardiac reconstitution. Significant improvement in left ventricular function, as evaluated by transthoracic echocardiography and micro-tip conductance catheter, was also observed. CONCLUSIONS AND SIGNIFICANCE: Skeletal muscle-derived multipotent Sk-34 cells that can give rise to skeletal and smooth muscle cells as reported previously, also give rise to cardiac muscle cells as multi-myogenic stem cells, and thus are a potential source for practical cellular cardiomyoplasty

Plasma levels of matrix metalloproteinase‐9 (MMP‐9) are associated with cognitive performance in patients with schizophrenia

Aim: Matrix metalloproteinase‐9 (MMP‐9) has been shown to modulate synaptic plasticity and may contribute to the pathophysiology of schizophrenia. This study investigated the peripheral levels of MMP‐9 and its association with cognitive functions in patients with schizophrenia to see the possible involvement of MMP‐9 in pathophysiology of schizophrenia, especially in cognitive decline. Methods: We measured the plasma levels of MMP‐9 in 257 healthy controls and 249 patients with schizophrenia, including antipsychotic drug–free patients. We also explored the possible association between plasma MMP‐9 levels and cognitive performance in healthy controls and patients with schizophrenia using the Wechsler Adult Intelligence Scale, Third Edition (WAIS‐ III), the Wechsler Memory Scale‐Revised (WMS‐R), and the Rey Auditory Verbal Learning Test (AVLT). Results: We found that the plasma levels of MMP‐9 were significantly higher in patients with schizophrenia, including antipsychotic drug–free patients, than in healthy controls. We found a significant negative association between plasma MMP‐9 levels and cognitive performance in controls and patients with schizophrenia. Conclusion: Together, these convergent data suggest a possible biological mechanism for schizophrenia, whereby increased MMP‐9 levels are associated with cognitive impairment

Plasma Levels of Soluble Tumor Necrosis Factor Receptor 2 (sTNFR2) Are Associated with Hippocampal Volume and Cognitive Performance in Patients with Schizophrenia

Background: An imbalance in the inflammatory tumor necrosis factor system, including soluble tumor necrosis factor receptor 2 (sTNFR2), may contribute to the pathophysiology of schizophrenia. Methods: We measured the plasma levels of sTNFR2 in 256 healthy controls and 250 patients with schizophrenia including antipsychotic drug-free patients and treatment-resistant patients. We also explored the possible association between plasma sTNFR2 levels and cognitive performance in healthy controls and patients with schizophrenia using the Wechsler Adult Intelligence Scale, Third Edition, the Wechsler Memory Scale-Revised, and the Rey Auditory Verbal Learning Test. An association between plasma sTNFR2 levels and hippocampal volume in controls and patients with schizophrenia was also investigated via MRI. Results: We found that the plasma levels of sTNFR2 were significantly higher in patients with schizophrenia, including both antipsychotic drug-free patients and treatment-resistant patients. We found a significant negative association between plasma sTNFR2 levels and cognitive performance in controls and patients with schizophrenia. Hippocampal volume was also negatively associated with plasma sTNFR2 levels in patients with schizophrenia. Conclusion: Together, these convergent data suggest a possible biological mechanism for schizophrenia, whereby increased sTNFR2 levels are associated with a smaller hippocampal volume and cognitive impairment

Mild Heat Stress Affects on the Cell Wall Structure in Candida albicans Biofilm

福岡歯科大学2018年

Implementasi manajemen sarana dan prasarana dalam meningkatkan mutu pendidikan pada madrasah tsanawiyah negeri (MTSN) Rantauprapat Kabupaten Labuhanbatu

Penelitian ini bertujuan untuk mengetahui perencanaan, pengorganisasian, pelaksanaan, pengawasan manajemen sarana dan prasarana dalam meningkatkan mutu pendidikan pada Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu. Penelitian ini dilakukan dengan menggunakan pendekatan kualitatif, Teknik pengumpulan data dalam penelitian ini menggunakan observasi, wawancara dan dokumen. Data yang didapat kemudian dianalisis dengan menggunakan analisis data kualitatif yang terdiri dari: (a).reduksi data, (b).penyajian data, dan (c) penarikan kesimpulan. Temuan penelitian: (1) perencanaan manajemen sarana dan prasarana di Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu terlebih dahulu dilakukan analisis kebutuhan riil baik yang menyangkut kebutuhan administrasi maupun pendukung kegiatan proses pembelajaran, seperti ruang kelas, moubilair, dan lain sebagainya. Yang melibatkan: Kepala Madrasah, KTU, bendahara, PKM, dan bahkan utusan dari komite sekolah. (2).Pengorganisasian manajemen sarana dan prasarana pada Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu dilakukan berdasarkan rumpun (kelompok) dari setiap jenis sarana itu sendiri, misalnya: bangunan fisik, moubilair, ATK, lingkungan, dan lain sebagainya yang kesemuanya itu di arsiparis berdasarkan ketentuan yang berlaku. (3) Pelaksanaan manajemen sarana dan prasarana di Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu berjalan baik dan lancar. Pelaksanaannya masing-masing pihak bekerja sesuai job/pekerjaan masing-masing dan sesuai kepentingannya, sehingga sistem kerja tidak ada tumpang tindih antara satu sama lain. Dan pertanggung jawabannya langsung kepada Kepala madrasah MTsN Rantauprapat walaupun tetap di bawah koordinasi PKM sarana dan prasarana. (4).Pengawasan manajemen sarana dan prasarana pada Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu dilakukan dengan cara: a) Pengawasan rutin setiap harinya yang dilakukan oleh PKM sarana jika menyangkut persoalan sarana pendukung pembelajaran, sedangkan yang menyangkut administrasi dilakukan oleh KTU. b) Secara berkala yakni setiap 6 (enam) bulan sekali diadakan rapat evaluasi tentang keadaan sarana dan prasarana. (5) Terkait dengan evaluasi diketahui bahwa sarana dan prasarana di Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu sudah terpenuhi dan sesuai dengan standar pendidikan nasional.