Comparison of capacities to maintain hematopoietic stem cells among different types of stem cells derived from the placenta and umbilical cord

AbstractIntroductionCord blood is utilized as a useful source of cells for hematopoietic stem cell transplantation, but this can be problematic because there is a high rate of graft failure compared to when other graft sources are used. A previous study successfully avoided graft failure by simultaneously grafting cord blood and bone marrow mesenchymal stem cells (MSCs) that are considered to function in the hematopoietic stem cell niche of the bone marrow.Organs of the fetal life support system such as the placenta and umbilical cord, which are discarded after delivery, contain an abundance of MSCs as well as cells that function in the hematopoietic stem cell niche. By identifying and collecting such cells and subsequently co-transplanting them with cord blood, an improvement in graft survival can be anticipated.MethodsThree types of stem cells, amnion epithelial stem cells (AM-Epi), amnion mesenchymal stem cells (AM-Mes), and Wharton's jelly (WJ)-MSCs, all of which can be isolated and cultured from the placenta amnion or umbilical cord WJ, were investigated for the expression of hematopoietic stem cell niche markers and for their capabilities to maintain hematopoietic stem cells when co-cultured with cord blood hematopoietic stem cells.ResultsAll types of isolated cells showed profiles that met the MSC minimal criteria according to surface marker analysis. In addition, all cell types expressed the hematopoietic stem cell niche marker stromal cell-derived factor-1 (SDF-1) (in order: AM-Epi > WJ-MSCs ≫ AM-Mes), although the expression declined with further passaging.After 5 days of co-culturing with cord blood CD34+ cells, the percentages of CD34+, CD45− cells were: AM-Epi 37.8%, AM-Mes 38.8%, WJ-MSCs 27.3%, and fibroblasts 27.4%; and the number of CFU-GM colonies were: AM-Epi 255.5 ± 21.6, AM-Mes 246.3 ± 28.5, WJ-MSCs 118.3 ± 11.8, fibroblasts 147.8 ± 19.0, and NC 121.3 ± 6.5. Statistical analyses demonstrated that AM-Epi and AM-Mes produced significantly greater numbers of CFU-GM compared to WJ-MSC, fibroblasts, or NC (p < 0.05).ConclusionsThese findings indicated that cells derived from the fetal life support system such as AM-Epi and AM-Mes can be anticipated as potential cell sources for clinical application in cell therapies for the purpose of enhancing graft survival during hematopoietic stem cell transplantation

Titanium single electron transistor fabricated by electron-beam lithography

A new method to fabricate non-superconducting mesoscopic tunnel junctions by oxidation of Ti is presented. The fabrication process uses conventional electron beam lithography and shadow deposition through an organic resist mask. Superconductivity in Ti is suppressed by performing the deposition under a suitable background pressure. We demonstrate the method by making a single electron transistor which operated at $T < 0.4$ K and had a moderate charge noise of $2.5 \times 10^{-3}$ e/$\sqrt{\mathrm{Hz}}$ at 10 Hz. Based on nonlinearities in the current-voltage characteristics at higher voltages, we deduce the oxide barrier height of approximately 110 mV.Comment: 6 pages, 4 figure

Hypervascularized bronchial arteries affect lung cancer surgery

Background: The present study investigated whether highly vascularized bronchial arteries affect the intraoperative blood loss and the operative time of video-assisted thoracic surgery (VATS) lobectomy for patients with non-small cell lung cancer. Methods: We retrospectively collected data on consecutive pathological stage I to IIIA non-small cell lung cancer patients who underwent VATS lobectomy with systematic lymph node dissection between January 2017 and December 2019. Patients were divided into the following two groups according to bronchial artery diameters on preoperative enhanced contrast computed tomography (CT) findings: ≤2 and >2 mm groups. Results: Among the 175 patients enrolled, risk factors for intraoperative blood loss >50 mL were being male (P=0.005), a history of smoking (P=0.01), percent forced expiratory volume in 1 s (FEV1.0%) 2.0 mm (P2.0 mm (P200 min were being male (P2.0 mm (P2.0 mm (P=0.024), and experience of surgeon <10 years (P=0.047) in the multivariable analysis. Conclusions: Bronchial artery diameter was the most important risk factor of intraoperative bleeding and prolonged operative time during VATS lobectomy

〔研究ノート〕ヒト血清アルブミンの糖化反応による カルボキシメチルリジンの生成箇所の同定

　　Human serum albumin（HSA）is an abundant plasma protein and is modified by glucose in plasma. The incubation of HSA with glucose at 37℃ for four weeks resulted in the formation of carboxymethyllysine in 11 sites of lysine residues of HSA（65, 190 . . .Ⅰ, 199, 233, 313, 378 . . .Ⅱ, 402, 432, 519, 525, 573 . . . Ⅲ）. The same experiments without glucose showed no carboxymethyllysine formation.　　Although early studies established that four lysine residues（Lys199, Lys281, Lys439 and Lys525）had been modified by nonenzymatic glycosylation, carboxymethylations of Lys281 and Lys439 were not detected in this experiment. Most carboxymethylation of lysine residues in HSA was located in domain Ⅲ in our study

Impact of surgical adhesion barrier on significant adhesion during a repeat cesarean section

　Background: Post-cesarean adhesions are associated with delayed infant delivery and infertility. In this retrospective study, we analyzed the effects of hyaluronic acid- carboxymethylcellulose (HA/CMC) membranes on postoperative adhesion during cesarean section.　Methods: Sixty-seven patients were divided into the surgical adhesion barrier used (n = 28) and not-used group (n = 39). We compared the severity of adhesion at the repeat cesarean section, as well the following variables: operation and incision delivery time, blood loss, and postoperative infection between both groups. The severity of adhesion was analyzed using the Zühike’s adhesion score between the abdominal wall and uterine corpus, and the Steinleitner’s uterine adhesion score.　Results: We found that the Zühike’s adhesion score between the abdominal wall and corpus of the uterus in the surgical adhesion barrier used group was significantly lower than that of the not-used group (0.46 ± 0.2 and 1.0 ± 0.2, respectively) (p = 0.04). The Steinleitner’s uterine adhesion score of the surgical adhesion barrier used group was lower than that of the not-used group (0.5 ± 0.3 and 1.3 ± 0.3, respectively), but not significantly (p = 0.07).　Discussion: We concluded that the surgical adhesion barrier was effective in preventing postoperative adhesion formation during cesarean sections. However, further investigations are necessary to reveal the usefulness of the surgical adhesion barrier during cesarean section, including its cost-effectiveness

マウスiPS細胞由来の気管支肺胞幹細胞は末梢気道上皮再生を促進する

Background: Bronchioalveolar stem cells (BASCs) located at the bronchioalveolar-duct junction (BADJ) are stem cells residing in alveoli and terminal bronchioles that can self-renew and differentiate into alveolar type (AT)-1 cells, AT-2 cells, club cells, and ciliated cells. Following terminal-bronchiole injury, BASCs increase in number and promote repair. However, whether BASCs can be differentiated from mouse-induced pluripotent stem cells (iPSCs) remains unreported, and the therapeutic potential of such cells is unclear. We therefore sought to differentiate BASCs from iPSCs and examine their potential for use in the treatment of epithelial injury in terminal bronchioles. Methods: BASCs were induced using a modified protocol for differentiating mouse iPSCs into AT-2 cells. Differentiated iPSCs were intratracheally transplanted into naphthalene-treated mice. The engraftment of BASCs into the BADJ and their subsequent ability to promote repair of injury to the airway epithelium were evaluated. Results: Flow cytometric analysis revealed that BASCs represented ~ 7% of the cells obtained. Additionally, ultrastructural analysis of these iPSC-derived BASCs via transmission electron microscopy showed that the cells containing secretory granules harboured microvilli, as well as small and immature lamellar body-like structures. When the differentiated iPSCs were intratracheally transplanted in naphthalene-induced airway epithelium injury, transplanted BASCs were found to be engrafted in the BADJ epithelium and alveolar spaces for 14 days after transplantation and to maintain the BASC phenotype. Notably, repair of the terminal-bronchiole epithelium was markedly promoted after transplantation of the differentiated iPSCs. Conclusions: Mouse iPSCs could be differentiated in vitro into cells that display a similar phenotype to BASCs. Given that the differentiated iPSCs promoted epithelial repair in the mouse model of naphthalene-induced airway epithelium injury, this method may serve as a basis for the development of treatments for terminal-bronchiole/alveolar-region disorders

CLE for visceral pleural invasion

Background: Visceral pleural invasion (VPI) in lung cancer is a significant prognostic factor; however, it is difficult to diagnose preoperatively or intraoperatively. In this study, we examined the possibility of intraoperative diagnosis of VPI using confocal laser endomicroscopy (CLE). Methods: Among patients with primary lung cancer who underwent surgery between April 2018 and August 2019, those in whom the tumor was in contact with the pleura on chest computed tomography and whose pleural changes were intraoperatively confirmed were enrolled in this study. In the 35 patients who underwent lung resection (6 cases with visceral pleural infiltration), the area where pleural change was noted was observed and a short video was recorded using CLE. Based on the video images, three evaluators determined the defect ratio (0%, 25%, 50%, 75%, and 100%) of the autofluorescence-positive structure. The area under the receiver operating characteristic curve was used to evaluate the diagnostic performance for VPI. In 15 cases (3 cases with VPI), a validation study was performed for intraoperative VPI according to the cutoff value of the defect ratio of the autofluorescence-positive structure. Results: The areas under the receiver operating characteristic curve for the defect ratio of the autofluorescence-positive structure were 0.86–0.91 for the three readers. Using defect ratio of autofluorescence-positive structure cutoff of ≥50% as predictor of VPI, the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 83.3–100.0%, 57.7–73.1%, 35.3–41.7%, 95.0–100.0%, and 75.0–78.1%, respectively, for the three readers. In the validation study, the sensitivity was 100%, the specificity was 83.3%, and the diagnostic accuracy rate was 86.7%. Conclusions: The diagnosis of VPI through CLE is simple, non-invasive, and has high diagnostic accuracy rates. This method may be applicable for determining surgical procedures

Quantitative Study of Neurons with Intracytoplasmic Pigments in Dorsal Root Ganglia Atomic Bomb and Aging

Three females in their fifties and five males in their seventies, all of whom had been exposed to the Nagasaki atomic bomb explosion in 1945, were investigated concerning the aging of neurons of the dorsal root ganglia. Through this study dealing with the frequency of neurons with intracytoplasmic pigments, lipofuscin, neuromelanin, eosinophilic granules and neurons free of pigment, no significant difference in four kinds of neurons between the exposed persons and the nonexposed persons was detected in 50-59 year-old females. Contrariwise in the 70-79 year-old males, the frequency, as average but not as individual persons, was significantly higher in lipofuscin, lower in neuromelanin, and unchanged in eosinophilic granules and no pigment

ユウロウセイ ノウキョウ ニタイシテ EWS オ モチイタ キカンシ ジュウテンジュツ ト ロウコウナイ フィブリンコ チュウニュウ ノ ヘイヨウ ガユウコウ デアッタ 1レイ

Background : Empyema with a bronchial fistula is difficult to treat. Recently, bronchial occlusion using endobronchial Watanabe Spigot（EWS）ris reported to be useful for treatment of intractable pneumothorax and thoracic empyema. Case : A ６０ year old man presented fever and left chest pain. He was diagnosed with empyema. Video－ assisted thoracoscopic debridement and decortication for the empyema cavity and drainage for the abscess cavity were performed. Air leak appeared at postoperative day１３. We performed EWS embolization and intrapleural administration of fibrin glue, and the persistent air leak disappeared. Conclusion : We experienced empyema with a bronchial fistula successfully treated with EWS embolization and intrapleural administration of fibrin glue