In utero transplantation of myoblasts and adipose-derived mesenchymal stem cells to murine models of Duchenne muscular dystrophy does not lead to engraftment and frequently results in fetal death.

Introduction: Duchenne muscular dystrophy (DMD) is a progressive disease that leads to damage of muscle and myocardium due to genetic abnormalities in the dystrophin gene. In utero cell transplantation that might facilitate allogenic transplantation is worth considering to treat this disease. Methods: We performed allogeneic in utero transplantation of GFP-positive myoblasts and adipose-derived mesenchymal stem cells into murine DMD model animals. The transplantation route in this study was fetal intraperitoneal transplantation and transplacental transplantation. Transplanted animals were examined at 4-weeks old by immunofluorescence staining and RT-qPCR. Results: No GFP-positive cells were found by immunofluorescence staining of skeletal muscle and no GFP mRNA was detected by RT-qPCR in any animal, transplantation method and cell type. Compared with previous reports, myoblast transplantation exhibited an equivalent mortality rate, but adipose-derived stem cell (ASC) transplantation produced a higher mortality rate. Conclusions: In utero transplantation of myoblasts or ASCs to murine models of DMD does not lead to engraftment and, in ASC transplantation primarily, frequently results in fetal death.

Laminin-221-derived recombinant fragment facilitates isolation of cultured skeletal myoblasts.

Introduction: Laminin is a major component of the basement membrane, containing multiple domains that bind integrin, collagen, nidogen, dystroglycan, and heparan sulfate. Laminin-221, expressed in skeletal and cardiac muscles, has strong affinity for the cell-surface receptor, integrin α7X2ß1. The E8 domain of laminin-221, which is essential for cell integrin binding, is commercially available as a purified recombinant protein fragment. In this study, recombinant E8 fragment was used to purify primary rodent myoblasts. We established a facile and inexpensive method for primary myoblast culture exploiting the high affinity binding of integrin α7X2ß1 to laminin-221. Methods: Total cell populations from dissociated muscle tissue were enzymatically digested and seeded onto laminin-221 E8 fragment-coated dishes. The culture medium containing non-adherent floating cells was removed after 2-hour culture at 37 °C. The adherent cells were subjected to immunofluorescence staining of desmin, differentiation experiments, and gene expression analysis. Results: The cells obtained were 70.3 ± 5.49% (n = 5) desmin positive in mouse and 67.7 ± 1.65% (n = 3) in rat. Immunofluorescent staining and gene expression analyses of cultured cells showed phenotypic traits of myoblasts. Conclusion: This study reports a novel facile method for primary culture of myoblasts obtained from mouse and rat skeletal muscle by exploiting the high affinity of integrin α7X2ß1 to laminin-221.

Comparison between hemodynamic effects of propofol and thiopental during general anesthesia induction with remifentanil infusion: a double-blind, age-stratified, randomized study.

PURPOSE: Propofol is commonly used with remifentanil for induction of general anesthesia (GA); however, it often leads to hypotension. Intraoperative hypotension is associated with postoperative adverse events. By contrast, thiopental has less negative inotropic effects on hemodynamics compared to propofol, which could be suitable to prevent hypotension during GA induction. In the present age-stratified, randomized, assessor-blinded study, using the ClearSight® system, we compared the hemodynamic effects of propofol and thiopental during GA induction under remifentanil infusion in non-cardiac surgery. METHODS: Patients were divided into young (20-40 year), middle (41-70 year), and elderly (> 70 year) groups (n = 20, each group). General anesthesia was induced with remifentanil 0.3 µg/kg/min, followed by propofol (2.0, 1.5, and 1.2 mg/kg) or thiopental (5.0, 4.0, and 3.0 mg/kg) in the young, middle, and elderly groups, respectively. The primary outcome was the difference in the decrease in mean arterial blood pressure between patients receiving propofol and thiopental in each age group. The secondary outcomes included other hemodynamic parameters and minimal bispectral index values measured up to 10 min after tracheal intubation. RESULTS: The decrease in mean arterial blood pressure was greater in patients receiving propofol than those receiving thiopental (- 45.4 vs - 26.6 mmHg and - 45.7 vs - 28.9 mmHg, P = 0.003 and 0.007, respectively), whereas no significant difference was observed in the young age group (P = 0.96). CONCLUSIONS: Thiopental is a more suitable agent than propofol for avoiding hypotension during GA induction under remifentanil infusion in the middle and elderly patients.

A case of esophageal perforation after intraoperative transesophageal echocardiography in a patient with a giant left atrium: unexpectedly large distortion of the esophagus revealed on retrospectively constructed three-dimensional imaging.

BACKGROUND: Esophageal perforation is a rare but serious complication of transesophageal echocardiography (TEE). An enlarged left atrium (LA), which is commonly associated with mitral stenosis (MS), is an under-recognized risk factor for esophageal perforation after intraoperative TEE. CASE PRESENTATION: We describe a case of TEE-induced esophageal perforation after cardiac surgery in a 79-year-old woman with a giant LA due to MS. Esophageal perforation was detected on postoperative day 6. After surgical repair, the patient gradually recovered with prolonged conservative treatment. Retrospectively constructed three-dimensional chest computed tomography images revealed an unusually distorted esophagus that was possibly vulnerable to injury. CONCLUSION: A giant LA can markedly distort the esophagus. It should be recognized as a risk factor for TEE-induced esophageal perforation.

Novel virus discovery in field-collected mosquito larvae using an improved system for rapid determination of viral RNA sequences (RDV ver4.0).

In this study, we improved a method for rapid determination of viral RNA sequences (RDV) to overcome the limitations of previous versions. The RDV ver4.0 method can detect RNA sequences with at least 1,000 copies as starting material. A novel virus, which was isolated from field-collected Aedes aegypti larvae in the Phasi Charoen district of Thailand using C6/36 cells, was identified using the RDV ver4.0 protocol. The virus was named Phasi Charoen virus (PhaV). We used a high-throughput pyrosequencing approach to obtain more information about the genome sequence of PhaV. Analysis of a phylogenic tree based on amino acid sequences strongly suggested that PhaV belongs to the family Bunyaviridae.

Surgically resected hepatocellular carcinomas in patients with non-alcoholic steatohepatitis.

BACKGROUND/AIMS: Surgically resected hepatocellular carcinomas (HCC) in patients with non-alcoholic steatohepatitis (NASH) have rarely been described and the clinicopathological characteristics of HCC and non-cancerous liver tissue are still obscure. METHODOLOGY: From 1997 to 2004, 242 patients with HCC underwent hepatic resection at the Hiroshima Red Cross Hospital and Atomic Bomb Survivors' Hospital. Among this group, the diagnosis of NASH was made in 3 patients. RESULTS: All 3 patients with HCC had cirrhosis. The tumor cells contained Mallory bodies and fat. The non-cancerous areas showed nodular regeneration with fatty changes, ballooning degeneration, and mild inflammatory infiltrates, as well as perivenular and perisinusoidal fibrosis. CONCLUSIONS: Patients with NASH and cirrhosis may progress to HCC, and careful follow-up based on tumor markers and imaging modalities, is essential to detect resectable HCC in patients with NASH and cirrhosis.

Rapid determination of viral RNA sequences in mosquitoes collected in the field.

A method for rapid determination of viral RNA sequences (RDV) was applied to homogenates of Aedes aegypti collected in Thailand in an area in which dengue fever (dengue hemorrhagic fever) is endemic, using the mosquito cell line C6/36. Nucleic acid sequences of dengue virus type 4 and cell fusing agent virus were detected. This RDV method has the potential to become a standard method for detection of both known and newly emerging, unknown mosquito-borne viruses.