17α-ethinylestradiol modulates endothelial function in ovariectomized rat carotid arteries.

17α-ethinylestradiol (EE2), a derivative of 17ß-estradiol (E2), is a potent estrogenic substance that is used as the estrogenic component of oral contraceptives (OCPs). However, women who take OCPs have an increased risk of cardiovascular events. Since few studies have examined EE2 endothelial effects, we explored the effects of EE2 on endothelial function in ovariectomized and isoflavone-free rats. After ovariectomy, 12-week-old female Sprague-Dawley rats were assigned to EE2, E2 or control groups. After 16 weeks, the EE2 and E2 groups were orally administered EE2 (8.3 µg/day) and E2 (12.6 µg/day) for 4 weeks, respectively. At 18 weeks, endothelial denudation of the left common carotid arteries was performed, and they were harvested at 20 weeks. The rats in the EE2 and E2 groups exhibited significantly decreased body weights and significantly increased uterine weights, respectively, but no differences were observed between the EE2 and E2 groups. The EE2 and E2 groups showed significantly enhanced acetylcholine-induced endothelium-dependent relaxation, with apamin plus charybdotoxin inhibiting only the EE2 group. Endothelial nitric oxide (NO) synthase expression was significantly higher in the EE2 group than in the control, but lower than in the E2 group. The intima-to-media ratio of denuded arteries was significantly lower in the E2 group than in the other groups, suggesting that NO decreased in the EE2 group compared to the E2 group. We conclude that EE2 has a weaker ability than E2 to produce NO and, for the first time, we demonstrate the ability of EE2 to enhance the activity of endothelial-derived hyperpolarizing factor.

Restoration of keratinocytic phenotypes in autonomous trisomy-rescued cells.

BACKGROUND: An extra copy of chromosome 21 in humans can alter cellular phenotypes as well as immune and metabolic systems. Down syndrome is associated with many health-related problems and age-related disorders including dermatological abnormalities. However, few studies have focused on the impact of trisomy 21 (T21) on epidermal stem cells and progenitor cell dysfunction. Here, we investigated the differences in keratinocytic characteristics between Down syndrome and euploid cells by differentiating cells from trisomy 21-induced pluripotent stem cells (T21-iPSCs) and autonomous rescued disomy 21-iPSCs (D21-iPSCs). METHODS: Our protocol for keratinocytic differentiation of T21-iPSCs and D21-iPSCs was employed. For propagation of T21- and D21-iPSC-derived keratinocytes and cell sheet formation, the culture medium supplemented with Rho kinase inhibitor on mouse feeder cells was introduced as growth rate decreased. Before passaging, selection of a keratinocytic population with differential dispase reactivity was performed. Three-dimensional (3D) air-liquid interface was performed in order to evaluate the ability of iPSC-derived keratinocytes to differentiate and form stratified squamous epithelium. RESULTS: Trisomy-rescued disomy 21-iPSCs were capable of epidermal differentiation and expressed keratinocytic markers such as KRT14 and TP63 upon differentiation compared to trisomy 21-iPSCs. The lifespan of iPSC-derived keratinocytes could successfully be extended on mouse feeder cells in media containing Rho kinase inhibitor, to more than 34 population doublings over a period of 160 days. Dispase-based purification of disomy iPSC-derived keratinocytes contributed epidermal sheet formation. The trisomy-rescued disomy 21-iPSC-derived keratinocytes with an expanded lifespan generated 3D skin in combination with a dermal fibroblast component. CONCLUSIONS: Keratinocytes derived from autonomous trisomy-rescued iPSC have the ability of stratification for manufacturing 3D skin with restoration of keratinocytic functions.

Reductive Amination of Ketonic Compounds Catalyzed by Cp*Ir(III) Complexes Bearing a Picolinamidato Ligand.

Cp*Ir complexes bearing a 2-picolinamide moiety serve as effective catalysts for the direct reductive amination of ketonic compounds to give primary amines under transfer hydrogenation conditions using ammonium formate as both the nitrogen and hydrogen source. The clean and operationally simple transformation proceeds with a substrate to catalyst molar ratio (S/C) of up to 20,000 at relatively low temperature and exhibits excellent chemoselectivity toward primary amines.

Reproductive Interference in an Introduced Bumblebee: Polyandry may Mitigate Negative Reproductive Impact.

As a signature of reproductive interference (RI), we reviewed hybrid production in eusocial bumblebees in Japan, by comparing introduced Bombus terrestris with native B. ignitus in Honshu (main island of Japan) and with native B. hypocrita sapporoensis in Hokkaido (northern island of Japan). In this review, we present additional new data showing hybrid production between introduced B. terrestris and native B. ignitus in Honshu. Interspecific mating with introduced B. terrestris disrupts the reproduction of native B. h. sapporoensis and B. ignitus, which belong to the same subgenus of Bombus, through inviable egg production. This interference appears to facilitate species replacement on Hokkaido. Simultaneously, the mating frequencies for queens of B. terrestris have increased, suggesting that polyandry might evolve in response to the extent of RI between B. terrestris and B. h. sapporoensis. To suppress the population size of B. terrestris in Hokkaido, two methods have been proposed: the mass release of B. h. sapporoensis males to induce RI between the two species and the spraying of insecticides against foraging workers so that the workers will carry the insecticides back to their colonies, killing the immature bees within the colonies. A candidate insecticide type is insect growth regulator, which may disrupt larval development without any apparent effect on foraging workers.

Autonomous trisomic rescue of Down syndrome cells.

Down syndrome is the most frequent chromosomal abnormality among live-born infants. All Down syndrome patients have mental retardation and are prone to develop early onset Alzheimer's disease. However, it has not yet been elucidated whether there is a correlation between the phenotype of Down syndrome and the extra chromosome 21. In this study, we continuously cultivated induced pluripotent stem cells (iPSCs) with chromosome 21 trisomy for more than 70 weeks, and serendipitously obtained revertant cells with normal chromosome 21 diploids from the trisomic cells during long-term cultivation. Repeated experiments revealed that this trisomy rescue was not due to mosaicism of chromosome 21 diploid cells and occurred at an extremely high frequency. We herewith report the spontaneous correction from chromosome 21 trisomy to disomy without genetic manipulation, chemical treatment or exposure to irradiation. The revertant diploid cells will possibly serve a reference for drug screening and a raw material of regenerative medicinal products for cell-based therapy.

Laparoscopic sigmoidectomy in a case of sigmoid colon cancer with situs inversus totalis.

Situs inversus totalis (SIT) is a rare anatomic anomaly in which organs in the chest and abdomen exist in a mirror image reversal of their normal positions. SIT can complicate surgical procedures, and few reports have described laparoscopic surgery for colorectal cancer in patients with SIT. Here, we report a case of successful laparoscopic surgery in a patient with SIT and sigmoid colon cancer. Laparoscopic sigmoidectomy involved colonic mobilization with high ligation of the inferior mesenteric vessels and complete mesocolic excision. The operating surgeon stood on the patient's left side, opposite the normal location for sigmoidectomy. By placing a 12-mm trocar in the left iliac fossa and using an automatic endoscopic linear stapler, the operating surgeon was able to perform left-handed colon resection without having to change position or move the laparoscopic monitor mid-procedure. An automatic endoscopic linear stapler is useful for laparoscopic left-side colon surgery in a patient with SIT.

[A Case of Anal Canal Cancer with Skin Invasion to Which Abdominoperineal Resection and Perineum Reconstruction Were Performed after Neoadjuvant Chemoradiotherapy].

A 74-year-old man visited our hospital with an awareness of anal mass and bleeding. He was diagnosed as adenocarcino- ma of anal canal with wide spreading skin invasion. After neoadjuvant chemoradiotherapy(radiationtotal 45 Gy/25 Fr; cape- citabine 825mg/m2)was performed to reduce the mass volume, laparoscopic abdominoperineal resection with large perineum skin resection and lateral lymph node dissection was carried out. The perineal defect was repaired with a rectus abdominis musculocutaneous flap. Six days after surgery, the rectus abdominis musculocutaneous flap necrotized, and second perineum reconstruction by the bilateral gracilis musculocutaneous flaps was performed after debridement of necrotic tissue. We reported a case of radical resection of local advanced anal canal cancer with skin invasion by performing combined modality therapy and perineum reconstruction.

Krüppel-like factor 4 expression in oral carcinoma cells and hypermethylation at the gene promoter.

BACKGROUND: Krüppel-like factor 4 (KLF4) is a transcription factor regulating proliferation-differentiation balance of epithelium, and down-regulated in less-differentiated and advanced oral carcinomas. Although the expression is inactivated by the promoter hypermethylation in malignant tumor cells, it remains unknown in oral carcinoma cells. METHODS: Genomic DNA isolated from nine different oral carcinoma cell lines and a normal keratinocyte line was treated with sodium bisulfite, and methylation at KLF4 gene promoter was determined by PCR direct-sequence analysis. KLF4 expression in cells cultured with or without demethylation reagent was monitored by quantitative real-time PCR and immunoblot. RESULTS: A 237-bp promoter region spanning - 718 and - 482 of KLF4 gene was hypermethylated in oral carcinoma cells that express KLF4 at a low level, but the methylation was infrequent in cells expressing KLF4 high amount. The downstream region from - 481 to +192 was not methylated in any cell lines. Demethylation treatment of cells up-regulated the expression at mRNA and protein levels. CONCLUSION: This study demonstrated that hypermethylation at a narrow range of the promoter region down-regulates KLF4 expression, and suggests that the loss of expression by the hypermethylation contributes to oral carcinoma progression.

Stability of lenalidomide suspension after preparation by a simple suspension method for enteral tube administration.

PURPOSE: A simple suspension method has been developed for tube administration, in which tablets (and capsules) are disintegrated in hot water (55℃) without grinding (or opening) them. In the present study, we evaluated the feasibility of this simple suspension method for the preparation of lenalidomide (Celgene, Summit, New Jersey and USA) suspension by testing the stability of this drug at 55℃ and its adsorbability on the tube. METHODS: We examined, by high-performance liquid chromatography, the time-dependent changes in the concentration of lenalidomide in suspensions of the drug prepared by the simple suspension method. The high-performance liquid chromatography analyses of lenalidomide were performed on Prominence LC-20AB/SPD-20 A (Shimadzu, Kyoto, Japan) with a ZORBAX SB-C18 RR analytical column (Agilent Technologies, Santa Clara, California, USA; particle size: 2.1 × 100 mm, 3.5 µm) at a flow rate of 0.4 mL/min. A solvent system consisting of 10 mM ammonium acetate (pH 7.0)/acetonitrile was used as the eluent and the eluate was detected by UV at 254 nm. RESULTS: Lenalidomide was confirmed to remain stable in hot water at 55℃ for 24 h in the prepared suspension by the simple suspension method, and more than 99% of the drug could be recovered from the suspension. In addition, 94.5-98.0% of the drug amount could pass through a percutaneous endoscopic gastrostomy tube. Lenalidomide was scarcely adsorbed on to the percutaneous endoscopic gastrostomy tube made of polyurethane or polyvinyl chloride. CONCLUSION: Lenalidomide was found to be stable even in hot water and was not adsorbed on to the percutaneous endoscopic gastrostomy tube.

Tactile search for change has less memory than visual search for change.

Haptic perception of a 2D image is thought to make heavy demands on working memory. During active exploration, humans need to store the latest local sensory information and integrate it with kinesthetic information from hand and finger locations in order to generate a coherent perception. This tactile integration has not been studied as extensively as visual shape integration. In the current study, we compared working-memory capacity for tactile exploration to that of visual exploration as measured in change-detection tasks. We found smaller memory capacity during tactile exploration (approximately 1 item) compared with visual exploration (2-10 items). These differences generalized to position memory and could not be attributed to insufficient stimulus-exposure durations, acuity differences between modalities, or uncertainty over the position of items. This low capacity for tactile memory suggests that the haptic system is almost amnesic when outside the fingertips and that there is little or no cross-position integration.

Gamma-polyglutamic acid-coated vectors for effective and safe gene therapy.

In the present study, we developed some novel gene delivery vectors, coated cationic complexes with gamma-polyglutamic acid (gamma-PGA) for effective and safe gene therapy. Cationic complexes were constructed with pDNA and cationic vectors, such as poly-L-arginine hydrochloride (PLA), poly-L-lysine hydrobromide (PLL), N-[1-(2, 3-dioleyloxy) propyl]-N, N, N-trimethylammonium chloride (DOTMA)-cholesterol (Chol) liposomes, and DOTMA-dioleylphosphatidylethanolamine (DOPE) liposomes. The cationic complexes showed high gene expression with strong cytotoxicity in melanoma B16-F10 cells. The cationic complexes were also strongly toxic to erythrocytes. On the other hand, the gamma-PGA was able to coat all cationic complexes and form stable nano-sized particles with negative charges. These gamma-PGA-coated complexes had high gene expression without cytotoxicity and toxicities to the erythrocytes. In in vivo transfection experiments, polyplexes showed high transfection efficiency over 10(5) RLU/g in the lung tissue after intravenous injection, although gamma-PGA-coated polyplexes showed a high value in the spleen. High transfection efficiency in lipoplexes and gamma-PGA-coated lipoplexes was observed in the spleen and lung. Thus, gamma-PGA-coated vectors are useful for clinical gene therapy.

Bone marrow stromal cells and bone marrow-derived mononuclear cells: which are suitable as cell source of transplantation for mice infarct brain?

There are few studies that denote whether bone marrow stromal cells (BMSC) and bone marrow-derived mononuclear cells (MNC) show the same therapeutic effects, when directly transplanted into the infarct brain. This study therefore aimed to compare their biological properties and behaviors in the infarct brain. Mouse BMSC were harvested and cultured. Mouse MNC were obtained through centrifugation techniques. Their cell markers were analyzed with FACS analysis. The MNC (10(6) cells; n = 10) or BMSC (2 x 10(5) cells; n = 10) were stereotactically transplanted into the ipsilateral striatum of the mice subjected to permanent middle cerebral artery occlusion at 7 days after the insult. Their survival, migration, and differentiation in the infarct brain were precisely analyzed using immunohistochemistry 4 weeks after transplantation. The MNC were positive for CD34, CD45, CD90, but were negative for Sca-1. The BMSC were positive for CD90 and Sca-1. The transplanted BMSC, but not MNC, extensively migrated into the peri-infarct area. Approximately 20% of the transplanted BMSC expressed a neuronal marker, NeuN in the infarct brain, although only 1.4% of the transplanted MNC expressed NeuN. These findings strongly suggest that there are large, biological differences between MNC and BMSC as cell sources of regenerative medicine for ischemic stroke.

Factors affecting performance of hospital infection control in Japan.

BACKGROUND: In Japan, hospital infection control (IC) programs are frequently underresourced, and their improvement is considered a pressing issue. METHODS: In 2005, we conducted a questionnaire survey of 638 teaching hospitals (most with 300 or more beds) and 882 nonteaching hospitals (most with fewer than 300 beds) in Japan. We analyzed associations among resources, infrastructures, activities, and performance related to IC. RESULTS: A total of 423 teaching hospitals (66.3%) and 377 nonteaching hospitals (50.2%) responded to the survey. The teaching hospitals had more IC infrastructure, such as full-time infection control practitioners (ICPs), link nurses, and infection control teams (ICTs), compared with the nonteaching hospitals. Infection surveillance was more likely to be implemented in hospitals with more ICP full-time equivalents (FTEs). IC performance scores were significantly higher in the teaching hospitals than in the nonteaching hospitals. In multivariate analyses, greater IC infrastructure, such as ICP FTEs, full-time IC nurses, and regular ICT rounds were significantly associated with IC performance. Hospital accreditation and hospital size also were significantly associated with higher IC performance scores. CONCLUSION: Given the strong associations found among IC infrastructure and performance, a new framework for evaluating IC infrastructure and for providing financial support may be effective in enhancing IC programs.

Impact of hospital accreditation on infection control programs in teaching hospitals in Japan.

BACKGROUND: In Japan, hospital infection control (IC) programs are frequently under-resourced, whereas their improvement is considered a pressing issue. Hospital accreditation may have a positive impact on IC program performance. The Japan Council for Quality Health Care (JCQHC) is a hospital accreditation organization that now prescribes broad elements of IC as part of its accreditation standards. METHODS: We sent questionnaire surveys to all teaching hospitals in Japan to characterize the current situation of hospital IC activities and identify the impact of accreditation on IC infrastructure and performance. The self-administered questionnaire that we used was developed based on the JCQHC accreditation standards. Surveys were sent to all institutions in 2004 and again in 2005. RESULTS: Of the 638 hospitals surveyed, 335 (52%) answered in both years. Most IC practitioners in Japanese teaching hospitals were working part time and spent limited hours performing IC duties. Surveillance was poorly implemented in Japan, and IC activities without evidence of effectiveness were widely performed. Surveillance was implemented more frequently in hospitals with adequate IC staffing. Improvement in IC infrastructure and performance between the surveys was larger in the newly accredited hospitals than the others. CONCLUSIONS: Hospital accreditation had a significant impact on hospitals' IC infrastructure and performance.

Asymmetric total synthesis of ent-sandaracopimaradiene, a biosynthetic intermediate of oryzalexins.

An asymmetric total synthesis of ent-sandaracopimaradiene, a biosynthetic intermediate of oryzalexins, via B-alkyl Suzuki-Miyaura coupling and Lewis acid-mediated B-ring formation as key steps was achieved.

[A life saving case of Vibrio vulnificus (septic type)].

After eating raw shellfish, 62-year-old man suffered from watery diarrhea and leg pain. The disease developed to septicemia and necrotizing fasciitis of bilateral lower extremities. Vibrio vulnificus was isolated from wound. He was treated with antibiotics, surgical debridement and hyperbaric oxygen therapy, and consequently was saved. Effect of hyperbaric oxygen therapy on V. vulnificus infection was discussed.