Lymphoid Lineage γδ T Cells Were Successfully Generated from Human Pluripotent Stem Cells via Hemogenic Endothelium

γδ T cells are a rare and unique prototype of T cells that share properties with natural killer cells in secondary lymphoid organs. Although many studies have revealed the function and importance of adult-derived γδ T cells in cancer biology and regenerative medicine, the low numbers of these cells hamper their application as therapeutic cell sources in the clinic. To solve this problem, pluripotent stem cell-derived γδ T cells are considered alternative cell sources; however, few studies have reported the generation of human pluripotent stem cell-derived γδ T cells. In the present study, we investigated whether lymphoid lineage γδ T cells were successfully generated from human pluripotent stem cells via hemogenic endothelium under defined culture conditions. Our results revealed that pluripotent stem cells successfully generated γδ T cells with an overall increase in transcriptional activity of lymphoid lineage genes and cytolytic factors, indicating the importance of the optimization of culture conditions in generating lymphoid lineage γδ T cells. We uncovered an initial step in differentiating γδ T cells that could be applied to basic and translational investigations in the field of cancer biology. Based on our result, we will develop an appropriate method to purify γδ T cells with functionality and it helpful for the study of basic mechanism of γδ T cells in pathophysiologic condition as well as clinic application.

Systematic Target Screening Revealed That Tif302 Could Be an OffTarget of the Antifungal Terbinafine in Fission Yeast

We used a heterozygous gene deletion library of fission yeasts comprising all essential and non-essential genes for a microarray screening of target genes of the antifungal terbinafine, which inhibits ergosterol synthesis via the erg1 enzyme. We identified 14 heterozygous strains corresponding to 10 non-essential [7 ribosomal-protein (RP) coding genes, spt7, spt20, and elp2] and 4 essential genes (tif302, rpl2501, rpl31, and erg1). Expectedly, their erg1 mRNA and protein levels had decreased compared to the control strain SP286. When we studied the action mechanism of the non-essential target genes using cognate haploid deletion strains, knockout of SAGA-subunit genes caused a down-regulation in erg1 transcription compared to the control strain ED668. However, knockout of RP genes conferred no susceptibility to ergosterol-targeting antifungals. Surprisingly, the RP genes participated in the erg1 transcription as components of repressor complexes as observed in a comparison analysis of the experimental ratio of erg1 mRNA. To understand the action mechanism of the interaction between the drug and the novel essential target genes, we performed isobologram assays with terbinafine and econazole (or cycloheximide). Terbinafine susceptibility of the tif302 heterozygous strain was attributed to both decreased erg1 mRNA levels and inhibition of translation. Moreover, Tif302 was required for efficacy of both terbinafine and cycloheximide. Based on a molecular modeling analysis, terbinafine could directly bind to Tif302 in yeasts, suggesting Tif302 as a potential off-target of terbinafine. In conclusion, this genome-wide screening system can be harnessed for the identification and characterization of target genes under any condition of interest.

Knockdown of vps54 aggravates tamoxifen-induced cytotoxicity in fission yeast

Tamoxifen (TAM) is an anticancer drug used to treat estrogen receptor (ER)‒positive breast cancer. However, its ER-independent cytotoxic and antifungal activities have prompted debates on its mechanism of action. To achieve a better understanding of the ER-independent antifungal action mechanisms of TAM, we systematically identified TAM-sensitive genes through microarray screening of the heterozygous gene deletion library in fission yeast (Schizosaccharomyces pombe). Secondary confirmation was followed by a spotting assay, finally yielding 13 TAM-sensitive genes under the drug-induced haploinsufficient condition. For these 13 TAM-sensitive genes, we conducted a comparative analysis of their Gene Ontology (GO) ‘biological process’ terms identified from other genome-wide screenings of the budding yeast deletion library and the MCF7breast cancer cell line. Several TAM-sensitive genes overlapped between the yeast strains and MCF7 in GO terms including ‘cell cycle’ (cdc2, rik1, pas1, and leo1), ‘signaling’ (sck2, oga1, and cki3), and ‘vesicle-mediated transport’ (SPCC126.08c, vps54, sec72, and tvp15), suggesting their roles in the ER-independent cytotoxic effects of TAM. We recently reported that the cki3 gene with the ‘signaling’ GO term was related to the ER-independent antifungal action mechanisms of TAM in yeast. In this study, we report that haploinsufficiency of the essential vps54 gene, which encodes the GARP complex subunit, significantly aggravated TAM sensitivity and led to an enlarged vesicle structure in comparison with the SP286 control strain. These results strongly suggest that the vesicle-mediated transport process might be another action mechanism of the ER-independent antifungal or cytotoxic effects of TAM.

Comparison of crown designs of different dental occupational groups, using CAD-CAM / 대한치과보철학회지

PURPOSE: Increasing use of computer aided design-computer aided manufacturing (CAD-CAM) system and number of design software made design of restoration easy and quick. Outcome of restoration has been dependent on dental technician's wax up proficiency, dentists can design restoration for themselves now. This study aims to investigate the outcome of restoration designs, according to handling skill of CAD-CAM design tool. MATERIALS AND METHODS: A patient's mandibular right 1st molar was prepared. After taking impression, stone model was made, scanned the stone model with 3 shape intra-oral scanner, stereolithography (STL) file was extracted. With 3shape dental designer, one dental technician with more than 5 years work experience (designer 0) and three dental technicians with less than 2years work experience (designer 1, 2, 3-group DT) and 4 1st year residents (designer 4, 5, 6, 7-group RT) designed gold crown on the same STL file. Designed crown's MD (mesio-distal) and BL (bucco-lingual) diameter, height of crown, inter-cuspal distance, number of occlusal contact points were compared. Statistical analysis was carried out, test of normality within each group, using independent t-test. Number of contact points were compared, using Wilcoxon signed-rank test. RESULTS: There was no significant difference between group DT and group RT. Number of contact points also resulted in no significant difference. CONCLUSION: The outcome of each designed crowns showed no statistical differences, in values which can be expressed as numbers. Subjective factors were different. With increasing proficiency in handling designing software, fabrication of restorations according to each designer's occlusal concept can be made easy.

The effect of copper alloy scaler tip on the surface roughness of dental implant and restorative materials / 대한치과보철학회지

PURPOSE: This study is designed to investigate the various impacts of different types of scaler tips such as cooper alloy base tip and the others on the surface roughness of teeth and implant by the method which is currently in clinical use. MATERIALS AND METHODS: Four different types of disc shaped porcelain, titanium, zirconia, and Type III gold alloy dental materials sized 15 mm diameter, 1.5 mm thickness were used for the experiment. Plastic hand curette (Group PS), cooper alloy new tip (Group IS), and stainless steel tip (Group SS) were used as testing appliances. A total of 64 specimens were used for this study; Four specimens for each material and appliance group. Surface roughness was formed with 15 degree angle in ultrasonic scaler tip and with 45 degree angle in hand curette of instrument tip and the specimen surface with 5 mm long, one horizontal-reciprocating motion per second for 30 seconds by 40 g force. To survey the surface roughness of each specimen, a field emission scanning electron microscope, an atomic force microscope, and a surface profiler were used. (Ra, microm). RESULTS: According to SEM, most increased surface roughness was observed in SS group while IS groups had minimal roughness change. Measurement by atomic force microscope presented that the surface roughness of SS group was significantly greater than those of PS, IS and control groups in the type III gold alloy group (P<.05). IS group showed lesser surface roughness changes compared to SS group in porcelain and gold alloy group (P<.05). According to surface profiler, surface roughness of SS group showed greater than those of PS, IS and control groups and IS group showed lesser than those of SS group in all specimen groups. Type III gold alloy group had large changes on surface roughness than those of porcelain, titanium, zirconia (P<.05). CONCLUSION: The result of this study showed that newly developed copper alloy scaler tip can cause minimal roughness impacts on the surface of implant and dental materials; therefore this may be a useful alternative for prophylaxis of implant and restored teeth.

Quality of Life Differences between Younger and Older Breast Cancer Patients / 한국유방암학회지

PURPOSE: This study was designed to investigate quality of life (QOL) differences between younger ( or =50 years) breast cancer survivors and to determine the unique effect of age on QOL after adjusting age-correlated variables. METHODS: One thousand two hundred fifty patients were enrolled. Clinicopatholgical and social parameters were reviewed and Functional Assessment of Cancer Therapy-Breast cancer instrument (FACT-B) and the Ladder of Life scale were used to measure the QOL. Among 1,250 eligible patients, 1,094 patients completed the questionnaire and were used for analysis. Chi-square test, t-test and a series of multiple regression analyses were conducted to verify age-related differences in QOL between two groups and to evaluate the unique contribution of age variable on QOL of breast cancer patients. RESULTS: Significant socio-demographic and clinical differences existed based on age categories, including education, job, time since surgery, chemotherapy and daily activity. Also, there were significant age-related differences in FACT-B total, physical well-being, social/family well-being, functional well-being and breast cancer subscale and in subjective QOL. Older patients > or =50 years showed significantly lower QOL than younger patients <50 years. However, after controlling for age-correlated variables including job, education, time since surgery, chemotherapy, and daily activity, there was no unique age difference in QOL among breast cancer survivors. CONCLUSION: Our study results suggest that older women with breast carcinoma suffered significantly lower QOL, even though the unique age effect was not found. Therefore, various interventions for enhancing QOL for women with breast carcinoma should be provided to older age group.