Time- and cell-specific activation of BMP signaling restrains chondrocyte hypertrophy.

Stem cell therapies for degenerative cartilage disease are limited by an incomplete understanding of hyaline cartilage formation and maintenance. Human bone marrow stromal cells/skeletal stem cells (hBMSCs/SSCs) produce stable hyaline cartilage when attached to hyaluronic acid-coated fibrin microbeads (HyA-FMBs), yet the mechanism remains unclear. In vitro, hBMSC/SSC/HyA-FMB organoids exhibited reduced BMP signaling early in chondrogenic differentiation, followed by restoration of BMP signaling in chondrogenic IGFBP5 + /MGP + cells. Subsequently, human-induced pluripotent stem cell (hiPSC)-derived sclerotome cells were established (BMP inhibition) and then treated with transforming growth factor ß (TGF-ß) -/+ BMP2 and growth differentiation factor 5 (GDF5) (BMP signaling activation). TGF-ß alone elicited a weak chondrogenic response, but TGF-ß/BMP2/GDF5 led to delamination of SOX9 + aggregates (chondrospheroids) with high expression of COL2A1, ACAN, and PRG4 and minimal expression of COL10A1 and ALP in vitro. While transplanted hBMSCs/SSCs/HyA-FMBs did not heal articular cartilage defects in immunocompromised rodents, chondrospheroid-derived cells/HyA-FMBs formed non-hypertrophic cartilage that persisted until at least 5 months in vivo.

Discovery of a new primer set for detection and quantification of Ilyonectria mors-panacis in soils for ginseng cultivation.

BACKGROUND: Korean ginseng is an important cash crop in Asian countries. However, plant yield is reduced by pathogens. Among the Ilyonectria radicicola-species complex, I. mors-panacis is responsible for root-rot and replant failure of ginseng in Asia. The development of new methods to reveal the existence of the pathogen before cultivation is started is essential. Therefore, a quantitative real-time polymerase chain reaction method was developed to detect and quantify the pathogen in ginseng soils. METHODS: In this study, a species-specific histone H3 primer set was developed for the quantification of I. mors-panacis. The primer set was used on DNA from other microbes to evaluate its sensitivity and selectivity for I. mors-panacis DNA. Sterilized soil samples artificially infected with the pathogen at different concentrations were used to evaluate the ability of the primer set to detect the pathogen population in the soil DNA. Finally, the pathogen was quantified in many natural soil samples. RESULTS: The designed primer set was found to be sensitive and selective for I. mors-panacis DNA. In artificially infected sterilized soil samples, using quantitative real-time polymerase chain reaction the estimated amount of template was positively correlated with the pathogen concentration in soil samples (R 2  = 0.95), disease severity index (R 2  = 0.99), and colony-forming units (R 2  = 0.87). In natural soils, the pathogen was recorded in most fields producing bad yields at a range of 5.82 ± 2.35 pg/g to 892.34 ± 103.70 pg/g of soil. CONCLUSION: According to these results, the proposed primer set is applicable for estimating soil quality before ginseng cultivation. This will contribute to disease management and crop protection in the future.

Overexpression of peptidyl-prolyl isomerase Pin1 attenuates hepatocytes apoptosis and secondary necrosis following carbon tetrachloride-induced acute liver injury in mice.

Pin1, a member of the parvulin family of PPIase enzymes, plays a crucial role in the post phosphorylation regulation that governs important roles in the cell signaling mechanism and regulates a variety of cellular events. In this study, we investigated the role of Pin1 in carbon tetrachloride (CCl(4))-induced apoptosis and necrosis of hepatocytes during acute liver injury of mice. An in vivo study was done with the overexpression of Pin1 in the mouse liver; using Pin1-adenoviruse (ad-Pin1) followed by CCl(4) injection to induce acute liver injury. Pin1 overexpression in the liver of the experimental mice attenuated acute liver injury induced by CCl(4) . Serum aminotransferases and the number of apoptotic cells were decreased compared to those of control virus injected mice. In addition, Pin1 overexpression increased NF-kB activity, as evidenced by increased DNA binding. In conclusion, Pin1 reduces acute liver injury of mice due to CCl(4) by modulating apoptotic signals and by increasing NF-kB activity.

The establishment and characterization of immortal hepatocyte cell lines from a mouse liver injury model.

Hepatocytes are an important research tool used for numerous applications. However, a short life span and a limited capacity to replicate in vitro limit the usefulness of primary hepatocyte cultures. We have hypothesized that in vivo priming of hepatocyte could make them more susceptible to growth factors in the medium for continuous proliferation in vitro. Here, a novel approach used to establish hepatocyte cell lines that included hepatocyte priming in vivo prior to culture with a 3,5-diethoxycarbonyl-1,4-dihydrocollidine diet was attempted. The cell line grew in a monolayer while maintaining a granular cytoplasm and a round nucleus. Electron microscopy displayed hepatocyte-like features including mitochondria, glycogen granules, and the presence of bile canaliculi. This cell line expressed many mature hepatocyte-specific genes including albumin, alpha1-antitrypsin, glucose 6-phosphatase, and tyrosine aminotransferase. Functional characteristic of hepatocytes like the ability to store glycogen, lipid, and synthesis of urea is well demonstrated by this cell line. These cells demonstrated anchorage dependent growth properties in soft agar and did not form tumors after transplantation into nude mice. This cell line can be sustained in culture for more than 100 passages (>1.5 years) without undergoing noticeable morphological changes or transformation. This novel method resulted in the establishment of an immortal, non-transformed hepatocyte cell line with functional characteristics that may aid research of cell metabolism, toxicology, and hepatocyte transplantation.

Torsion of a mucocele of the vermiform appendix: a case report and review of the literature.

Torsion of a mucocele of the vermiform appendix is an extremely rare condition and also a rare cause of an acute abdomen with a clinical presentation that is indistinguishable from acute appendicitis, and thus, the condition is diagnosed during operation. Here, the authors describe the case of a 78-year-old female, who presented with intermittent abdominal pain. The appendix had a pelvic position and the torsion was counterclockwise. In addition, the torsion was associated with mucocele of the appendix, which was considered a secondary factor of torsion. Appendectomy and drainage were performed.

Choledochal cyst with ectopic distal location of the papilla of Vater.

In cholangiographic techniques, the close relationship between choledochal cyst and anomalous union of pancreaticobiliary duct has attracted medical attention. There have been rare cases in which the papilla of Vater was found in a position other than its normal position, and such cases have been reported sporadically. However, such cases are interesting in the anatomical context. In this review, we present our experience of choledochal cyst in a 30-month-old boy in whom the papilla of Vater was positioned in the third portion of the duodenum.

Carcinosarcoma of the rectosigmoid colon in a 13-year-old girl.

Reported herein is an unusual case of carcinosarcoma of the colon. A 13-year-old girl was transferred to Chonbuk National University Hospital, Korea with a known pelvic mass. CT and magnetic resonance imaging confirmed a large pelvic mass. A sarcoma was diagnosed following colonoscopic biopsy. An ultra-low anterior resection with pelvic lymph node dissection was performed. The tumor consisted of a well-differentiated adenocarcinoma showing strong immunoreactivity to epithelial markers (pancytokeratin, cytokeratin 7, cytokeratin 19, cytokeratin 20, epithelial membrane antigen, and CEA) and a sarcomatous lesion with strong diffuse vimentin expression but no immunoreactivity to any of the six epithelial markers. Carcinosarcomas of the colon are extremely rare, and all reported cases involve adults. To the authors' knowledge, this is the first reported case of a carcinosarcoma of the colon in a child.