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ABSTRACT Background Methods Results Conclusions Odontogenic keratocyst (OKC) is a partial manifestation of Gorlin syndrome (GS), resulting from the abnormal activation of the hedgehog signaling pathway. OKC predominantly occurs in young adults and is mostly asymptomatic at the time of initial diagnosis. As OKC is asymptomatic, GS can be challenging to diagnose in certain instances. In this study, we attempted to identify asymptomatic GS from sporadic OKC cases using a previously developed gene panel for GS.Genomic DNA was extracted from patient samples. These DNA samples were analyzed using the AmpliSeq Custom DNA Panel (Illumina), which was specifically designed to target four previously established genes (PTCH1, PTCH2, SMO, and SUFU). Mutations from patients were predicted using tools, such as MutationTaster, CADD, and Polyphen‐2.Thirty‐one patients with OKC were included: 22 sporadic, 9 syndromic, 14 cases with dentigerous cysts, and 3 patients with orthokeratinized odontogenic cysts. One patient with sporadic OKC carried 50% genetic mutation in the cyst and blood, indicative of GS. PTCH1 mutations were found in one of the 14 patients with dentigerous cysts, 3 of the 17 first‐time sporadic cases, and all four recurrent cases. Resected OKC tissues revealed a PTCH1 mutation.We found one patient with GS from those diagnosed with sporadic OKC. Our findings suggest that PTCH1 mutations are associated with postoperative recurrence of OKC, implying that hedgehog‐related gene variations may contribute to jaw cyst development and improve the prognosis of OKC. [ABSTRACT FROM AUTHOR]

Translocator protein (TSPO) is a mitochondrial outer membrane protein expressed on a variety of immune cells, including macrophages, dendritic cells, and T cells, in addition to neurons and steroid-producing cells. Previous studies of TSPO ligands have suggested that TSPO is involved in multiple cellular functions, including steroidogenesis, immunomodulation, and cell proliferation. Currently, there are limited reports on the effects of TSPO or TSPO ligands on T cell-mediated immune responses. We here investigated the involvement of TSPO/TSPO ligand in T cell responses using a 2,4-dinitro-1-fluorobenzene (DNFB)-induced contact hypersensitivity (CH) model. Treatment with Ro5-4864, a TSPO ligand, during DNFB sensitization reduced the number and activation status of CD4+ and CD8+ T cells in draining lymph nodes and alleviated skin inflammation after DNFB challenge. Adoptive transfer of Ro5-4864-treated mouse-derived DNFB-sensitized T cells to naïve mice inhibited CH responses after DNFB challenge. Ro5-4864-treated sensitized T cells showed lower proliferative responses when stimulated with DNFB-pulsed antigen-presenting cells compared to control-treated sensitized T cells. Ro5-4864 also suppressed cell proliferation, as well as adenosine triphosphate and lactate production, during T cell activation. Moreover, the inhibitory effects of Ro5-4864 on T cell responses were conserved in TSPO-deficient cells. Our results suggest that Ro5-4864 inhibits CH responses by suppressing energy metabolism, at least via glycolysis, to reduce the T cell primary response in a TSPO-independent manner., (© The Author(s) 2024. Published by Oxford University Press on behalf of The Japanese Society for Immunology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Titanium is a biocompatible material commonly used for dental treatments. However, the detailed mechanism underlying the weak biological activity of titanium has not been elucidated. We investigated both the inflammatory responses and T cell activation induced by solid titanium in the gingiva in mice. Both titanium and nickel wire implantation promoted neutrophil infiltration into the gingiva on day 2. Nickel, but not titanium, wire implantation enhanced proinflammatory cytokine expression and dendritic cell activity in gingival tissue by day 2. Nickel wire implantation enhanced the activity of T cells in draining lymph nodes on day 5. Moreover, T cell and neutrophil infiltration and elevated proinflammatory cytokine expression in the gingival tissue were still observed on day 5. However, no such augmented biological responses were observed after titanium wire implantation. These findings suggest that, unlike nickel, solid titanium does not induce sufficient inflammatory responses leading to T cell activation in gingival tissue.

We recently reported a new method to purify the induced pluripotent stem (iPS)-derived osteoprogenitors (iPSop). In this paper, we optimized the procedure and characterized cells at each process step. We observed that 10 days of treatment with FGF-2, IGF-1 and TGF-β (FIT) resulted in early-phase osteoblasts and 14 days of treatment resulted in late-phase osteoblasts. We found that treatment with 1,25(OH)2 vitamin D3 increased expression of osteocalcin and decreased expression of tissue-non-specific alkaline phosphatase and runt-related transcription factor 2 (RUNX2) in iPSop-day14 cells (cells treated with FIT for 14 days). Therefore, iPSop-day14 cells were promoted to mature osteoblasts by 1,25(OH)2 vitamin D3 treatment. In addition, we found that 1,25(OH)2 vitamin D3 treatment for 14 days enhanced not only mineralization but also expression of osteocyte markers, including dentin matrix protein-1 and fibroblast growth factor-23, in iPSop cells. Therefore, 1,25(OH)2 vitamin D3 is a potent promoter of osteoblast-osteocyte transition. The results of this study suggest that it is possible to evaluate both early- and late-phase osteoblasts and to apply cells to drug screening for anabolic drugs that stimulate bone formation.

We and others have reported that cancer side population (SP) cells have self-renewal and multidrug resistance capabilities. These phenotypes are similar to those of cancer stem cells (CSCs), cancer stem-like cells and tumor-initiating cells (TICs). It has also been reported that upregulation of the epidermal growth factor receptor (EGFR) significantly increases the number of cancer SP cells, conversely, molecular targeting of EGFR tyrosine kinases using specific kinase inhibitors downregulates CSCs. Thus, we used flow cytometric analysis and cell sorting to examine cancer SP cells in the SCA9.cl-15, WR21 and A253 cell lines that originate from a salivary gland tumor (SGT). We successfully isolated cancer SP cells from all of these cell lines. SP cells were detected following treatment of these cell lines with the receptor tyrosine kinase inhibitors (RTKIs) lapatinib, erlotinib and vandetanib. Several studies reported that RTKIs mostly reduced the SP population in cancer cells. We did not observe any detectable morphological differences between SP cells and non-SP cells. We found that the EGF RTKI lapatinib decreased the number of cancer SP cells in all cell lines investigated; however, the EGF RTKI erlotinib did not cause significant differences in the frequency of cancer SP cells in these cell lines. Addition of vandetanib significantly increased the number of cancer SP cells and upregulated the phosphorylated Akt. As far as we know, this is the first report to show that one of the RTKIs, vandetanib, can activate Akt and increase the number of cancer SP cells. It has been reported that RTKIs could competitively inhibit ABC transporters and subsequently reduced the number of SP cells. However, our observation indicated that signaling changes induced by RTKIs could even activate Akt and induce the SP population. Investigation of the SP phenotype of SGTs is important for the establishment of optimal cancer therapy.

The ability of the human fungal pathogen Candida albicans to undergo the morphological transition from a single yeast form to pseudohyphal and hyphal forms in response to various conditions is known to be an important for its virulence. Many studies have shown the pharmacological effects of resveratrol, a phytoalexin polyphenolic compound. In this study, we investigated the antifungal activity of resveratrol against C. albicans. Both yeast-form and mycelial growth of C. albicans were inhibited by resveratrol. In addition, normal filamentation of C. albicans was affected and yeast-to-hypha transition under serum-, pH-, and nutrient-induced hyphal growth conditions was impaired by resveratrol.

Side population (SP) cells are isolated from various tissues and cell lines based on the exclusion of DNA-binding dye Hoechst 33,342 and exhibit potent stem cell characteristics. There have been few previous reports of SP cells in head and neck cancer cell lines. Thus, we isolated SP cells from oral squamous cell carcinoma cell line, Ho-1-N-1. Ho-1-N-1 contained 3.0% SP cells. Ho-1-N-1 SP cells showed self-renewal capacity, generating both SP and non-SP cells. Next, we analyzed differentially expressed genes between Ho-1-N-1 SP and non-SP cells using GeneChip microarray and quantitative real-time RT-PCR. SP cells expressed high levels of ATP-binding cassette transporters with related multidrug resistance (MDR) genes. The expression of ABCB1 and ABCG2 were significantly up-regulated in Ho-1-N-1 SP cells. In addition, the expression of CFLAR, BCL2 and BCL2A1 which are associated with anti-apoptosis, were also significantly increased in the SP cells. Chemoresistance to anticancer agents, including 5-fluorouracil and carboplatin, were compared between Ho1-N-1 SP and non-SP cells using flow cytometry and tetrazolium salt microtiter plate assay. Ho-1-N-1 SP cells survived significantly longer and SP ratio remarkably increased after anticancer agent treatment compared to non-SP cells. Immunocytochemical staining and apoptosis assay validated these results, and suggested an anti-apoptotic potential for Ho-1-N-1 SP cells. Ho-1-N-1 SP cells survived with various agents which were not only probably due to high level expression of ABC transporters, but also anti-apoptotic proteins. These observations indicated that Ho-1-N-1 SP cells were MDR phenotype and should be the main target for effective cancer therapy.

We report here side population (SP) cells, a cancer stem cell enriched fraction from pancreatic cancer cell line, have enormous superior potential of the epithelial to mesenchymal transition (EMT), invasion, and metastasis. In an isolated SP cell culture, the cells rapidly expressed and up-regulated E-cadherin, an epithelial phenotypic marker, and the cells formed tightly contacted cell cluster, which is a representative epithelial phenotypic appearance. When the SP cells were incubated in the presence of TGF-beta, SP cells changed their shape into mesenchymal-like appearance including spindle shaped assembly. This alteration was associated with significant reduction of E-cadherin expression level. TGF-beta induced EMT-associated gene alteration such as reduction of E-cadherin mRNA and induction of Snail mRNA and matrixmetalloproteinase (MMP)-2 mRNA. Finally, SP cells exerted notable matrigel invasion activity in response to TGF-beta treatment, whereas MP cells did not respond to TGF-beta-mediated invasion. In conclusion, these results suggest that SP cells from pancreatic cancer cell line possess superior potentials of phenotypic switch, i.e., EMT/MET, micro-invasion, and in vivo metastasis, as compared to MP cells. Because micro-invasion and metastasis are key mechanisms of cancer malignant potential, SP cells would be the attractive target for preventing cancer progression., (Copyright 2008 UICC.)

Aim: Transforming growth factor-beta (TGF-beta) has dual activity in tumor cells. We studied the effect of TGF-beta on tumor-initiating cells (TICs), which are similar in self-renewal and differentiation features to normal adult stem cells., Methods: We used side population (SP) cells that exclude DNA binding dye Hoechst 33342 to obtain TICs, studied the differences in the kinetics of the SP cell response to TGF-beta treatment between hepatic tumor cell lines, and performed gene analysis., Results: SP cells from all cell lines have higher proliferative ability compared to non-SP cells and they are drug resistant. TGF-beta treatment increased the percentage of SP cells (%SP) and the survival rate; chemotherapeutic drug resistance developed only in K-251 SP cells. Gene analysis showed that TGF-beta up-regulated epidermal growth factor receptor (EGFR) only in K-251 cells. There were no EGFR mutations in K-251, which had been reported in lung cancer. Knockdown of Smad4 using the small-interfering RNA technique in K-251 cells inhibited EGFR overexpression and significantly decreased the %SP. In contrast, the JNK inhibitor had little effect on EGFR expression or the %SP., Conclusion: TGF-beta treatment of K-251 cells causes tumor progression and the anti-cancer drug resistant phenotype by increasing SP.

Retinoic acid (RA) is produced via two sequential reactions of retinol dehydrogenases (RDHs) and retinal dehydrogenases (RALDHs). We found that primary cultured mouse hepatocytes on a single collagen gel gradually lost hepatocyte specific morphology, and that another collagen gel overlay remarkably recovered it. The levels of albumin and liver-dominant expressed RDHs expression in hepatocytes paralleled their morphological change, decreased during single collagen gel culture, and were up-regulated by sequential collagen overlay. Quite similar to the expression changes, albumin and those RDHs' mRNA expression levels increased along with liver differentiation during pre- and postnatal liver development. Our data supports that all-trans and 9-cis RA, catalyzed by the RDHs, indeed play an important role in liver differentiation and regeneration.