Stimulating Macropinocytosis for Intracellular Nucleic Acid and Protein Delivery: A Combined Strategy with Membrane-Lytic Peptides to Facilitate Endosomal Escape

Delivery of biomacromolecules via endocytic pathways requires the efficient accumulation of cargo molecules into endosomes, followed by their release to the cytosol. We propose a unique intracellular delivery strategy for bioactive molecules using a new potent macropinocytosis-inducing peptide derived from stromal-derived factor 1α (SN21). This peptide allowed extracellular materials to enter cells through the activation of macropinocytosis. To provide the ability to release internalized cargoes from endosomes, we conjugated SN21 with membrane-lytic peptides. The combination of a macropinocytosis-inducing peptide and a membrane-lytic peptide successfully delivered functional siRNA and proteins, which include antibodies, Cre recombinase, and an artificial transcription regulator protein having a transcription activator-like effector (TALE) motif. This study shows the feasibility of combining the physiological stimulation of macropinocytosis with the physicochemical disruption of endosomes as a strategy for intracellular delivery

Basic life support training for single rescuers efficiently augments their willingness to make early emergency calls with no available help: A cross-over questionnaire survey

Background: The aim of this study was to investigate effects of basic life support (BLS) training on willingness of single rescuers to make emergency calls during out-of-hospital cardiac arrests (OHCAs) with no available help from others.Methods: A cross-over questionnaire survey was conducted with two questionnaires. Questionnaires were administered before and after two BLS courses in fire departments. One questionnaire included two scenarios which simulate OHCAs occurring in situations where help from other rescuers is available (Scenario-M) and not available (Scenario-S). The conventional BLS course was designed for multiple rescuers (Course-M), and the other was designed for single rescuers (Course-S).Results: Of 2,312 respondents, 2,218 (95.9%) answered all questions and were included in the analysis. Although both Course-M and Course-S significantly augmented willingness to make early emergency calls not only in Scenario-M but also in Scenario-S, the willingness for Scenario-M after training course was significantly higher in respondents of Course-S than in those of Course-M (odds ratio 1.706, 95% confidential interval 1.301-2.237). Multiple logistic regression analysis for Scenario-M disclosed that post training (adjusted odds ratio 11.6, 95% confidence interval 7.84-18.0), age (0.99, 0.98-0.99), male gender (1.77, 1.39-2.24), prior BLS experience of at least three times (1.46, 1.25-2.59), and time passed since most recent training during 3 years or less (1.80, 1.25-2.59) were independently associated with willingness to make early emergency calls and that type of BLS course was not independently associated with willingness. Therefore, both Course-M and Course-S similarly augmented willingness in Scenario-M. However, in multiple logistic regression analyses for Scenario-S, Course-S was independently associated with willingness to make early emergency calls in Scenario-S (1.26, 1.00-1.57), indicating that Course-S more efficiently augmented willingness. Moreover, post training (2.30, 1.86-2.83) and male gender (1.26, 1.02-1.57) were other independent factors associated with willingness in Scenario-S.Conclusions: BLS courses designed for single rescuers with no help available from others are likely to augment willingness to make early emergency calls more efficiently than conventional BLS courses designed for multiple rescuers. © 2014 Hirose et al

One-Pot Synthesis of 6-Substituted Amino-2,4-diaminopyrimidine Derivatives Using Ketene Dithioacetals with Amines and Guanidine Carbonate

6-Substituted amino-2,4-diaminopyrimidine derivatives were prepared by one-pot synthesis using ketene dithioacetals, amine compounds, and guanidine carbonate in pyridine. These pyrimidine products displayed blue fluorescence in the solid state

Phlegmonous Colitis after Cold Snare Polypectomy in an Immunosuppressed Patient: A Case Report

Introduction: Cold snare polypectomy (CSP) is a procedure with a low risk of complications. Here, we present our experience of a rare case of submucosal abscess following CSP in an immunosuppressed patient. Case Presentation: Seventy-eight-year-old man underwent CSP, developing a fever, chills, and right lower abdominal pain 8 days later. Ultrasound and computed tomography revealed wall thickening of the ascending colon, presenting as whitening and thickening of the same region, and excretion of pus was observed after biopsy. The diagnosis was made as phlegmonous colitis, for which antibiotic therapy was commenced. The patient was diagnosed with chronic myelomonocytic leukemia (CMML) during admission. We considered the following reasons as possible causes of infectious complications after CSP: (1) the patient had a highly immunosuppressed state with comorbidities such as CMML as well as diabetes mellitus and (2) disruption of the mucosal barrier occurred during endoscopic resection. Conclusion: Although CSP is generally considered safe, our case highlights the potential for serious complications in immunosuppressed patients. Therefore, the decision to perform CSP in such patients should be made with caution to avoid unnecessary interventions. In instances where treatment is essential, thorough bowel preparation and prophylactic antibiotic use may be necessary to mitigate the risks

DEVELOPING A CONSENSUS MAIKING GAME OF NIMBY-TYPED PUBLIC PLANNING

The purpose of our study is to develop a consensus making game of NIMBY typed public planning, and to examine whether this game can be applied for understanding the idea of "veil of ignorance" as a measure to facilitate consensus-making in a NIMBY-type situation. The policy of siting repositories for radioactive waste by the Fukushima nuclear accident became difficult to introduce because of disagreement of the residents. In order to facilitate risk communication between administrations and residents, we develop a game to seek consensus of the policy. Twelve players take either mayor\u27s or resident\u27s role of six communities. The mayors discuss and propose evaluative criterias for judging options of siting repository, and the residents discuss and evaluate the options with the criterias proposed by the mayors

小麦アレルギーのエピトープペプチドに対するモノクローナル抗体の作製

Monoclonal Antibodies were raised against SQQQ(Q)PPF peptide conjugated to Keyhole Limpet Hemocyanin, which has been identified as epitope peptide of wheat allergy. Three monoclonal antibodies, mAbs 3D, 4B, and 6F were obtained by screening with the peptide conjugated to bovine serum albumin. However, these mAbs did not react with the free peptide in a competitive enzyme-linked immunosorbent assay. On western analysis, about 14 and 17kDa proteins in the wheat globulin fraction were detected with these mAbs. It is reported that wheat proteins of about 14, 16kDa were reactive with sera of patients allergic to wheat and that a wheat globulin of about 19kDa has SQQQQQG sequence. It seems likely that the mAbs obtained here recognize the 19kDa protein related to wheat allergy

フクマク ハシュ オ トモナッタ コウド シンコウ イガン ニ タイスル TS 1 ニヨル ジュツゼン カガク リョウホウ ノ ユウヨウセイ

TS -１ is an oral anticancer agent developed by utilizing biochemical modulation. We used TS -１in neoadjuvant chemotherapy for a patient with highly advanced gastric cancer that was accompanied by peritoneal dissemination. This enabled us to resect tumor. This patient was a６０-year-old woman. Fluoroscopic upper gastrointestinal series revealed a circumferential, type４lesion extending from the middle of the corpus to the antrum. This was diagnosed by endoscopy as poorly differentiated adenocarcinoma. CT showed ascites, thickening of the gastric wall, and direct infiltration into the head of the pancreas. In endoscopy of the large bowel, a strawberry jelly-like elevation was detected at the ileum. This was diagnosed as poorly differentiated adenocarcinoma, and considered a metastatic lesion produced by dissemination. Chest CT showed a single metastasis in the upper lobe of the right lung. We gave her３cycles of combined TS-１and low-dose CDDP for neoadjuvant chemotherapy. On laparotomy, we found that there was no ascites, and miliary scars were present at several sites near the ascending colon. The antrum of the stomach firmly adhered to the head of the pancreas, and scarred. We judged that the tumor was resectable, and performed distal gastrectomy（D２）plus ileocecal resection（D２）． In histopathological examination, poorly differentiated adenocarcinoma was detected only on a part of the muscular layer in the lesser curvature and posterior wall of the corpus, and marked fibrosis was observed in the submucosal layer. The effect of chemotherapy was histologically evaluated as grade２. The tumor was diagnosed as poorly differentiated adenocarcinoma（por）, with muscularis propria（mp）, lymph invasion２（ly２），vein invasion ０（v０）and degree of lymph node metastasis２（+）［n２（+）］. Tubular adenocarcinoma was detected in a part of the submucosal layer of the ileum. The patient was alive with cancer as of２７months after operation

A High-Speed Congenic Strategy Using First-Wave Male Germ Cells

BACKGROUND: In laboratory mice and rats, congenic breeding is essential for analyzing the genes of interest on specific genetic backgrounds and for analyzing quantitative trait loci. However, in theory it takes about 3-4 years to achieve a strain carrying about 99% of the recipient genome at the tenth backcrossing (N10). Even with marker-assisted selection, the so-called 'speed congenic strategy', it takes more than a year at N4 or N5. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe a new high-speed congenic system using round spermatids retrieved from immature males (22-25 days of age). We applied the technique to three genetically modified strains of mice: transgenic (TG), knockin (KI) and N-ethyl-N-nitrosourea (ENU)-induced mutants. The donor mice had mixed genetic backgrounds of C57BL/6 (B6):DBA/2 or B6:129 strains. At each generation, males used for backcrossing were selected based on polymorphic marker analysis and their round spermatids were injected into B6 strain oocytes. Backcrossing was repeated until N4 or N5. For the TG and ENU-mutant strains, the N5 generation was achieved on days 188 and 190 and the proportion of B6-homozygous loci was 100% (74 markers) and 97.7% (172/176 markers), respectively. For the KI strain, N4 was achieved on day 151, all the 86 markers being B6-homozygous as early as on day 106 at N3. The carrier males at the final generation were all fertile and propagated the modified genes. Thus, three congenic strains were established through rapid generation turnover between 41 and 44 days. CONCLUSIONS/SIGNIFICANCE: This new high-speed breeding strategy enables us to produce congenic strains within about half a year. It should provide the fastest protocol for precise definition of the phenotypic effects of genes of interest on desired genetic backgrounds

チョメイナ ノウホウ ケイセイ オ トモナッタ カンサイボウ ガン ノ 1ジケンレイ

It is said that in hepatocellular carcinoma, necrosis is liable to occur in the center and the percentage of cystic formation is comparatively low, but as we have experienced one case of hepatocellular carcinoma associated with marked cystic formation, we reported it. The patient was a female aged７０years. Because general fatigue and anorexia occurred, Abdominal CT test was conducted and hepatic mass was pointed out. Contrast-CT test showed a tumor of６cm in diameter in the medial segment of the left lobe of the liver. As the border was stained with arterial phase, the center was not imaged and the tumor was diagnosed as vascular proliferating type hepatic tumor associated with marked cystic change. MRI test showed that the border of the tumor was lobular and part thereof was of septal structure. The tumor was diagnosed as hepatocellular carcinoma associated with bleeding in tumor, hepatic cystadenocarcinoma or hepatic sarcoma. Abdominal angiography showed not only marked vascular proliferation and tumor stain in the area of the middle hepatic artery but also early venous return, and the middle hepatic vein was clearly imaged. The tumor size was ５×５×４cm in size and was in contact with the middle hepatic vein but it did not invade the vein. Left hepatic lobectomy was performed. Histopathologically the center of the tumor was cystic with colliquative necrosis. The tumor had the trabecular structure and the tumor cell was consisted of clear cell. The tumor was thus diagnosed as poorly differentiated hepatocellular carcinoma. Postoperative course was good and the patient was discharged from our hospital on the１９th day after operation. At the moment when１９ months have passed since the operation, the patient is alive without any recurrence of carcinoma