Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method

Background: We investigated and compared the osteogenic potential and bone regeneration capacities of dedifferentiated fat cells (DFAT cells) and adipose-derived stem cells (ASCs). Method: We isolated DFAT cells and ASCs from GFP mice. DFAT cells were established by a new culture method using a mesh culture instead of a ceiling culture. The isolated DFAT cells and ASCs were incubated in osteogenic medium, then alizarin red staining, alkaline phosphatase (ALP) assays, and RT-PCR (for RUNX2, osteopontin, DLX5, osterix, and osteocalcin) were performed to evaluate the osteoblastic differentiation ability of both cell types in vitro. In vivo, the DFAT cells and ASCs were incubated in osteogenic medium for four weeks and seeded on collagen composite scaffolds, then implanted subcutaneously into the backs of mice. We then performed hematoxylin and eosin staining and immunostaining for GFP and osteocalcin. Results: The alizarin red-stained areas in DFAT cells showed weak calcification ability at two weeks, but high calcification ability at three weeks, similar to ASCs. The ALP levels of ASCs increased earlier than in DFAT cells and showed a significant difference (p Conclusion: DFAT cells are easily isolated from a small amount of adipose tissue and are readily expanded with high purity; thus, DFAT cells are applicable to many tissue-engineering strategies and cell-based therapies

Cast: a novel protein of the cytomatrix at the active zone of synapses that forms a ternary complex with RIM1 and munc13-1

The cytomatrix at the active zone (CAZ) has been implicated in defining the site of Ca2+-dependent exocytosis of neurotransmitter. We have identified here a novel CAZ protein of ∼120 kD from rat brain and named it CAST (CAZ-associated structural protein). CAST had no transmembrane segment, but had four coiled-coil domains and a putative COOH-terminal consensus motif for binding to PDZ domains. CAST was localized at the CAZ of conventional synapses of mouse brain. CAST bound directly RIM1 and indirectly Munc13-1, presumably through RIM1, forming a ternary complex. RIM1 and Munc13-1 are CAZ proteins implicated in Ca2+-dependent exocytosis of neurotansmitters. Bassoon, another CAZ protein, was also associated with this ternary complex. These results suggest that a network of protein–protein interactions among the CAZ proteins exists at the CAZ. At the early stages of synapse formation, CAST was expressed and partly colocalized with bassoon in the axon shaft and the growth cone. The vesicles immunoisolated by antibassoon antibody–coupled beads contained not only bassoon but also CAST and RIM1. These results suggest that these CAZ proteins are at least partly transported on the same vesicles during synapse formation

ヨウジ 4～7サイ ノ タイリョク ノ ソウゴウテキ ブンセキ キョウオウセイ オヨビ ヘイコウセイ ノ ハッタツ ト チノウ ノ カンケイ

As a part of synthetic analysis if the infant-physical fitness, (from 4 to 7 years old) I examined the relation of the development of physical cooperation and balance to the intelligence quotient. Following results were obtained : 1) The development of girls\u27 cooperation function is better than that of boys. 2) The devlopment of girls\u27 balance function is better than that of boys. 3) As to intelligence quotient, girls are more excellent than boys. (P<0.005) 4) In the relation of the physical cooperation and balance function to the intelligence, the functions of both boys and girls had some degree of relation. As to the matter of age, I found the relation of the intelligence quotient to the degree of maturity of functions was lager as children, both boys and girls, are younger

ヨウジ 4～7サイ ノ タイリョク ノ ソウゴウテキ ブンセキ キンリョク オヨビ ジュウナンセイ ノ ハッタツ ト チノウ ノ カンケイ

As a part of general analysis of the infant-physical fitness, (children from 4 to 7 years old) I examined the relation of the development of muscular, strength and pliabellty to the intelligence quotient. The results following are obtained : 1) The development of the boys\u27 muscular strength function is better than that of the girls\u27. (P<0.005) 2) The development of the girls\u27 pliability function is better than that of the boys\u27. ( P<0.01～P<0.005) 3) As to intelligence quotient, there was no significant difference between the boys\u27 and the girls\u27. 4) As to the relation of the muscular strength and pliability function to the intelligence quotieut both of the function of the boys\u27 and the girls\u27 had some degrees of relation. As to the matter of age, I found the relation of the intelligence to the degree of maturity of the functions was lager auording as children, both boys\u27 and girls\u27 are younger

ヨウジ 4～7サイ ノ タイリョク ノ ソウゴウテキ ブンセキ タイカク オヨビ シュッセイ ジュンイ ト ビンショウセイ キノウ ノ カンケイ

As a part of synthetic analysis of infant-physical fitness, (from 4 to 7 years old) the relationship of the physical characteristics and birth order to reaction time was exmained. Following results were obtained : 1) The development of older children\u27s physique was better than that of younger children (p<00.5-0.005). 2) In releation to the birth order and physique, boys of first birth were better than third birth (p<0.05-0.005); but girls of third birth were better than first birth (p<0.05-0.005). 3) In relation to the physique and speed of reaction time, side step test, standing reacton time, and response reaction time both third birth boys and third birth girls were better than first and second birth boys and girls. 4) In relation to the birth order and speed of reaction time, both third birth boys and third birth girls were better than first and second birth boys and girls (p<0.05-0.005)

Comprehensive genomic profiling for patients with chemotherapy‐naïve advanced cancer

Comprehensive genomic profiling (CGP) testing by next-generation sequencing has been introduced into clinical practice as part of precision cancer medicine to select effective targeted therapies. However, whether CGP testing at the time of first-line chemotherapy could be clinically useful is not clear. We conducted this single-center, prospective, observational study to investigate the feasibility of CGP testing for chemotherapy-naïve patients with stage III/IV gastrointestinal cancer, rare cancer, and cancer of unknown primary, using the FoundationOne® companion diagnostic (F1CDx) assay. The primary outcome was the detection rate of at least one actionable/druggable cancer genomic alteration. Actionable/druggable cancer genomic alterations were determined by the F1CDx report. An institutional molecular tumor board determined the molecular-based recommended therapies. A total of 197 patients were enrolled from October 2018 to June 2019. CGP success rate was 76.6% (151 of 197 patients), and median turnaround time was 19 days (range: 10-329 days). Actionable and druggable cancer genomic alterations were reported in 145 (73.6%) and 124 (62.9%) patients, respectively. The highest detection rate of druggable genomic alterations in gastrointestinal cancers was 80% in colorectal cancer (48 of 60 patients). Molecular-based recommended therapies were determined in 46 patients (23.4%). CGP testing would be a useful tool for the identification of a potentially effective first-line chemotherapy