Secreted factors from adipose tissue-derived mesenchymal stem cells suppress oxygen/glucose deprivation-induced cardiomyocyte cell death via furin/PCSK-like enzyme activity

AbstractClinical application of mesenchymal stem cells (MSCs) represents a potential novel therapy for currently intractable deteriorating diseases or traumatic injuries, including myocardial infarction. However, the molecular mechanisms of the therapeutic effects have not been precisely revealed. Herein, we report that conditioned media (CM) from rat adipose tissue-derived MSCs (ASCs) protected adult cardiomyocytes from oxygen/glucose deprivation (OGD)-induced cell death. We focused on furin/PCSK protease activity in ASC-CM because many therapeutic factors of MSCs and soluble cardioprotective factors include the PCSK cleavage site. We found that recombinant furin protected cardiomyocytes from OGD-induced cell death. The ASC-CM had potent furin/PCSK protease activity and the cardioprotective effect of the CM from ASCs in the OGD-assay was abolished by an inhibitor of the furin/PCSK-like enzyme. Microarray analysis and Western blot analysis showed PCSK5A, the secreted type of PCSK5, is the most abundantly secreted PCSK among 7 PCSK family members in ASC. Finally, knockdown of PCSK5A in ASCs decreased both the furin/PCSK protease activity and cardioprotective activity in the CM. These findings indicate an involvement of furin/PCSK-type protease(s) in the anti-ischemic activity of ASCs, and suggest a new mechanism of the therapeutic effect of MSCs

Association Between SLFN11 and Antitumor Activity of Trabectedin

Background/Aim: Trabectedin is a DNA-damaging agent and has been approved for the treatment of patients with advanced soft tissue sarcoma. Schlafen 11 (SLFN11) was identified as a dominant determinant of the response to DNA-damaging agents. The aim of the study was to clarify the association between SLFN11 expression and the antitumor activity of trabectedin. Materials and Methods: The antitumor activity of trabectedin was evaluated under different expression levels of SLFN11 regulated by RNA interference and CRISPR-Cas9 systems, and the combined antitumor activity of ataxia telangiectasia and Rad3-related protein kinase (ATR) inhibitor and trabectedin in sarcoma cell lines using in vitro a cell viability assay and in vivo xenograft models. Results: SLFN11-knockdown cell lines had a lower sensitivity to trabectedin, compared to parental cells. ATR inhibitor enhanced the antitumor activity of trabectedin in SLFN11-knockdown cells and in a SLFN11-knockout xenograft model. Conclusion: SLFN11 expression might be a key factor in the antitumor activity of trabectedin

A Radioimmunoassay for Rat Serum Corticosterone

A reliable radioimmunoassay for rat serum corticosterone has been developed. 25 μ1 of diluted serum (1:100) was assayed with a specific antiserum raised against corticosterone-21-hemisuccinate conjugated to bovine serum albumin. The within-assay and between-assay coefficients of variation were 7.1% and 13.9%, respectively. The mean serum corticosterone concentration was 65.3±6.0 ng/ml (n=10). The corticosterone level increased to 208.4 ±23.7 ng/ml after ACTH administration, and was suppressed to the limit of assay sensitivity after dexamethasone administration

Analysis of Human Insulin Analogues in Vitro, Using Gel Chromatographic Method

The incubation medium and incubated human pancreas were gel chromatographed on the Bio-Gel P-30 column after extraction with acid ethanol. The extracted immunoreactive insulin (IRI) was successfully separated into two peaks at the position of 6000 molecular weight region. These two peaks corresponded to those which were detected in human serum. These findings suggest that the two groups of insulin are directly secreted from human pancreatic tissue. But the incorporated [3H] leucine peak into acid-ethanol extractable protein did not elute out at the same position as each insulin peak. Therefore, the measurement of [3H] leucine incorporation into acid-ethanol extractable protein is not a good indicator to evaluate insulin biosynthesis

Successful laparoscopic resection of virilizing ovarian steroid cell tumor, not otherwise specified, in a 22-year-old woman: a case report and evaluation of the steroidogenic pathway

Objective: Ovarian steroid cell tumor (SCT) is a rare tumor with steroid-producing ability. We report a 22-year-old woman with secondary amenorrhea and hirsutism caused by an ovarian SCT-not otherwise specified (NOS), who underwent successfully laparoscopic resection of the tumor. Case report: A 22-year-old null gravida woman presented to a hospital, having amenorrhea for 18 months and increasing facial hair. Physical examination revealed obesity (body mass index, 37.3 kg/m2) with evident facial and trunk hair. Total and free serum testosterone, and dehydroepiandrosterone sulfate levels were found to be elevated. Levels of serum adrenocorticotropic hormone, gonadotropins, cortisol, aldosterone, and ovarian steroids were observed to be within reference intervals. Although polycystic ovaries were not found, a hyperechogenic solid tumor (3 cm) was detected on transvaginal ultrasonography. Laparoscopic resection of the tumor was performed. One month post-surgery, total and free testosterone levels were observed to have decreased, and menstruation resumed two months thereafter. The patient was histologically diagnosed with ovarian SCT-NOS. Expression of ovarian steroidogenic enzymes, which are related to hyperandrogenism, was observed. No disease recurrence has been reported for more than 5 years post-surgery

Distribution and Elimination of Insulin and C-peptide in a Benign Insulinoma Patient

The distribution and elimination of insulin and C-peptide was evaluated in a case of benign insulinoma, using the method of gel chromatography. The significant differences between total Immunoreactive insulin (IRI) level and the level of Peak I plus Peak II of IRI was noticed in splenic vein. This fact suggested that intermediate and/or abnormal IRI could be released from the tumor. In order to diagnose a hypoglycemic patient with completely normal IRI and CPR level in peripheral blood, the gel chromatographic separation of IRI from splenic and/or portal blood could be useful