Non-canonical NFKB signaling endows suppressive function through FOXP3-dependent regulatory T cell program.

Regulatory T cells (Tregs) play a central role in modulating adaptive immune responses in humans and mice. The precise biological role of non-canonical nuclear factor 'κ-light-chain-enhancer' of activated B cells (NFKB) signaling in human Tregs has yet to be fully elucidated. To gain insight into this process, a Treg-like cell line (MT-2) was genetically modified using CRISPR/Cas9. Interestingly, NFKB2 knockout MT-2 cells exhibited downregulation of FOXP3, while NFKB1 knockout did not. Additionally, mRNA expression of FOXP3-dependent molecules was significantly reduced in NFKB2 knockout MT-2 cells. To better understand the functional role of the NFKB signaling, the NFKB1/NFKB2 loci of human primary Tregs were genetically edited using CRISPR/Cas9. Similar to MT-2 cells, NFKB2 knockout human Tregs displayed significantly reduced FOXP3 expression. Furthermore, NFKB2 knockout human Tregs showed downregulation of FOXP3-dependent molecules and a diminished suppressive function compared to wild-type and NFKB1 knockout Tregs. These findings indicate that non-canonical NFKB signaling maintains a Treg-like phenotype and suppressive function in human Tregs through the FOXP3-dependent regulatory T cell program.

Breast Cancers Secreting Sialyl-fibronectin Are Less Likely to Cause Epithelial-mesenchymal Transition and Have Good Prognoses.

Background/Aim: Elevated blood fibronectin (FN) levels have been observed in various cancers; however, their significance is controversial. We measured sialyl-fibronectin (S-FN), a type of FN secreted by tumor cells in the blood, and investigated whether blood S-FN secretion is associated with cancer malignancy and recurrent metastases. Materials and Methods: We constructed an enzyme-linked immunosorbent assay (ELISA) system that recognizes S-FN as an antigen and measured the amount of S-FN secreted into the blood of 89 patients with breast tumors. The relationship between S-FN secretion and prognostic predictors was analyzed. Immunostaining was performed to identify the site of S-FN secretion in the breast tissue. Results: Among the 82 patients, 21 (25.6%, 21/82) and 61 (74.4%, 61/82) were blood S-FN-positive and S-FN-negative, respectively. Regarding prognostic predictors, blood S-FN-positive and S-FN-negative patients showed significant difference in locoregional recurrence (p=0.026), remote metastases (p=0.049), and histological margins (p=0.001). Locoregional recurrence was associated with positive histological margins in S-FN-positive patients. However, remote metastases were associated with N-factor and histological classification (HC) in S-FN-negative patients. Furthermore, S-FN particles were detected in the cytoplasm of breast cancer cells through immunostaining. After the onset of recurrent metastases, two S-FN-positive and six S-FN-negative patients received anticancer drug treatment; however, further progression was observed in five S-FN-negative patients. Conclusion: S-FN-positive patients are less likely to develop distant metastases, have a better prognosis, and may be less resistant to therapeutic agents than S-FN-negative patients, which contain many epithelial-mesenchymal transition cells.

Serum Sialyl Fibronectin Is an Indicator of Good Prognosis in Thyroid Cancer.

BACKGROUND/AIM: Sialyl-fibronectin (S-FN), a type of glycoprotein like Sialyl Lewisa and MAC1 thyroid cancer biomarkers, has been found to be expressed in thyroid cancer. In this study, we examined the usefulness of serum S-FN as a biomarker, as well as prognostic factor in various tumors including thyroid cancer. PATIENTS AND METHODS: Using the MoAb JT-95, an ELISA kit was created and S-FN levels in sera (blood S-FN) of a total of 182 cases were investigated. Analysis included 63 cases of thyroid cancer, 33 cases of thyroid benign tumors, 7 cases of parathyroid benign tumors, and 79 cases of breast cancer. RESULTS: The incidence of blood S-FN-positive cases was 24 (38.0%) in 63 examined patients with thyroid cancer. Out of 40 examined benign neck tumor cases, 16/40 (40%) were S-FN-positive. Out of 79 examined breast cancer cases, 20/79 (25.3%) were S-FN-positive, with significant differences between thyroid and breast cancer cases (p=0.007). Regarding the association of blood S-FN expression and prognosis in thyroid cancer, there was a significant difference between 39 blood S-FN-negative cases and 15 recurrent or metastatic cases in terms of progression and pathological factors: tumor size, lymph node metastasis, extra-membrane infiltration, and clinical stage. All 63 assessed thyroid cancer cases also had a significant difference in these factors. There were no significant differences between these factors in the 24 blood S-FN-positive thyroid cancer cases. In cases of recurrent metastasis, intravascular cells were observed in all recurrent metastasis patients of both groups. Regarding staining of intravascular infiltrating cells, recurrent metastasis appeared at a higher rate in cases with S-FN-negative infiltrating cells. CONCLUSION: S-FN presence in blood can be used as an indicator of good prognosis in thyroid cancer patients.

Mice with R2509C-RYR1 mutation exhibit dysfunctional Ca2+ dynamics in primary skeletal myocytes.

Type 1 ryanodine receptor (RYR1) is a Ca2+ release channel in the sarcoplasmic reticulum (SR) of the skeletal muscle and plays a critical role in excitation-contraction coupling. Mutations in RYR1 cause severe muscle diseases, such as malignant hyperthermia, a disorder of Ca2+-induced Ca2+ release (CICR) through RYR1 from the SR. We recently reported that volatile anesthetics induce malignant hyperthermia (MH)-like episodes through enhanced CICR in heterozygous R2509C-RYR1 mice. However, the characterization of Ca2+ dynamics has yet to be investigated in skeletal muscle cells from homozygous mice because these animals die in utero. In the present study, we generated primary cultured skeletal myocytes from R2509C-RYR1 mice. No differences in cellular morphology were detected between wild type (WT) and mutant myocytes. Spontaneous Ca2+ transients and cellular contractions occurred in WT and heterozygous myocytes, but not in homozygous myocytes. Electron microscopic observation revealed that the sarcomere length was shortened to ∼1.7 µm in homozygous myocytes, as compared to ∼2.2 and ∼2.3 µm in WT and heterozygous myocytes, respectively. Consistently, the resting intracellular Ca2+ concentration was higher in homozygous myocytes than in WT or heterozygous myocytes, which may be coupled with a reduced Ca2+ concentration in the SR. Finally, using infrared laser-based microheating, we found that heterozygous myocytes showed larger heat-induced Ca2+ transients than WT myocytes. Our findings suggest that the R2509C mutation in RYR1 causes dysfunctional Ca2+ dynamics in a mutant-gene dose-dependent manner in the skeletal muscles, in turn provoking MH-like episodes and embryonic lethality in heterozygous and homozygous mice, respectively.

UV Sterilization Effects and Osteoblast Proliferation on Amorphous Carbon Films Classified Based on Optical Constants.

Optical classification methods that distinguish amorphous carbon films into six types based on refractive index and extinction coefficient have garnered increasing attention. In this study, five types of amorphous carbon films were prepared on Si substrates using different plasma processes, including physical and chemical vapor deposition. The refractive index and extinction coefficient of the amorphous carbon films were measured using spectroscopic ellipsometry, and the samples were classified into five amorphous carbon types-amorphous, hydrogenated amorphous, tetrahedral amorphous, polymer-like, and graphite-like carbon-based on optical constants. Each amorphous carbon type was irradiated with 253.7 nm UV treatment; the structure and surface properties of each were investigated before and after UV treatment. No significant changes were observed in film structure nor surface oxidation after UV sterilization progressed at approximately the same level for all amorphous carbon types. Osteoblast proliferation associated with amorphous carbon types was evaluated in vitro. Graphite-like carbon, which has relatively high surface oxidation levels, was associated with higher osteoblast proliferation levels than the other carbon types. Our findings inform the selection of suitable amorphous carbon types based on optical constants for use in specific medical devices related to osteoblasts, such as artificial joints and dental implants.

Correlation of Cell Proliferation with Surface Properties of Polymer-like Carbon Films of Different Thicknesses Prepared by a Radio-Frequency Plasma CVD Process.

In this study, correlation of cell proliferation with surface properties of the polymer-like carbon (PLC) films of different thicknesses prepared by radio-frequency plasma CVD are investigated. Four PLC samples were prepared via radio frequency plasma chemical vapor deposition on Si substrates. Each PLC film was analyzed using spectroscopic ellipsometry to determine its thickness, refractive index (n), and extinction coefficient (k); the thickness ranged from 29.0 to 356.5 nm. Based on their n−k plots, all the samples were classified as PLC-type films. The biological response of the PLC films was evaluated in vitro using a cell culture. The samples with relatively thick PLC films (>300 nm) exhibited stronger cell proliferation properties than those with thinner films. Moreover, the results of the surface analysis showed no significant differences in the surface composition of those PLC samples, as analyzed using X-ray photoelectron spectroscopy, but that as the PLC films became thicker, their surfaces became rougher on the nanoscale and their wettability improved. Overall, this study showed that careful control of the film growth of PLC films, which affects their surface properties, is essential for their use in bio-interface applications.

Effect of Thymidine Phosphorylase Gene Demethylation on Sensitivity to 5-Fluorouracil in Colorectal Cancer Cells.

BACKGROUND/AIM: Chemotherapy is used for recurrent and metastatic colorectal cancer, but the response rate of 5-fluorouracil (5-FU), the standard treatment for colorectal cancer, is low. We hypothesized that thymidine phosphorylase (TYMP) expression, a rate-limiting activating enzyme of 5-FU, is regulated by methylation of the gene promoter region, and demethylation of TYMP would increase sensitivity to 5-FU. MATERIALS AND METHODS: HCT116 colon cancer cells were treated with 5-aza-2'-deoxycytidine, a demethylating agent, and changes in TYMP transcription and sensitivity to 5-FU were evaluated. RESULTS: TYMP expression increased over 54-fold in HCT116 transfected with TYMP. The cytotoxicity of 5-FU increased up to 5.5-fold. In comparison, in HCT116 treated with 5-aza-2'-deoxycytidine, TYMP expression increased 5.8-fold. However, the cytotoxicity of 5-FU remained unchanged. CONCLUSION: Demethylating agent alone did not promote the cytotoxicity of 5-FU against colorectal cancer. To further increase the sensitivity to 5-FU, combination with adjuvant therapy focusing on metabolic pathways other than the TYMP pathway appear necessary.

Osteoblastic MC3T3-E1 cells on diamond-like carbon-coated silicon plates: Field emission scanning electron microscopy data.

Diamond-like carbon (DLC) is an amorphous form of carbon that contains aspects of both the diamond and graphite structures. It is composed of carbon and hydrogen, and owing to its texture, high mechanical hardness, chemical inertness, and optical transparency, DLC is widely used as a protective coating in the form of a thin film, which is applied to the surfaces of many materials. Recently, it has attracted attention as a biomedical material because of its high biocompatibility and stability [1,2]. DLC is particularly suitable to be embedded in the body owing to its low friction properties and selective cell surface attachment properties [3]. The material is currently being developed for the treatment of bone fractures [4]. However, unlike fibroblasts, the attachment of osteoblasts has not been extensively examined and no morphological data is available on how osteoblastic cells form contacts with the surface of biocompatible DLC-coated materials. Herein, such data were collected by coating DLC on the surface of silicon plates. The attachment of mouse cells to the DLC-coated plates was examined by colorimetric cell proliferation assay, and morphological observations were made using a field emission scanning electron microscope. Also, the flat cross section of the cell and plate was obtained by the ion milling method.

Gelatin Sponge as an Anchorage for Three-dimensional Culture of Colorectal Cancer Cells.

BACKGROUND: Compared to two-dimensional cultures, three-dimensional (3D) cultures have many advantages in cancer studies. Nevertheless, their implementation is unsatisfactory. This study aimed to develop an anchorage-dependent 3D culture model for colorectal cancer research. MATERIALS AND METHODS: Human HCT116, DLD-1 and SW620 colorectal cell lines were cultured in a gelatin sponge, and its applicability for morphological examination was studied. RESULTS: The resulting specimens were suitable for scanning electron microscopy, transmission electron microscopy, and immunohistochemical examination. HCT116 formed smaller structures and migrated through the pores of the sponge. DLD-1 formed larger structures with tight cell-to-cell adhesion. SW620 also formed large structures but small clustered cells tended to attach to the anchorage more favorably. Immunohistochemical staining demonstrated phosphorylated yes-associated protein (YAP) localized near the attachment site in HCT116 cells. CONCLUSION: Because the gelatin sponge provided suitable anchorage and the cultured cells formed distinguishable 3D structures, this method may be useful for further colorectal cancer research.

The Contrasting Perceptions and the Cause Regarding Patenting Technologies Between Academic Medical Researchers and Pharmaceutical Companies Based in Japan.

BACKGROUND: The recent trend of pharmaceutical companies commercializing new objects as new drugs based on the findings of academic medical researchers, commonly categorizing them as "academic drug discovery" is increasingly gaining popularity in the pharmaceutical industry. Studies state that academic researchers based in universities have lower motivation to apply for patents. However, none of the studies evaluated the existence and extent of the "motivation for patent" in academic researchers, being lower than that of pharmaceutical companies. This study assesses two hypotheses; H1: academic medical researchers are less likely to believe that the patent system is necessary for pharmaceuticals, and thus have diminished interest in the commercialization of their research findings when compared to those in the pharmaceutical industry, H2: apprehension of the raison d'être of the patent system affects positive impressions on patents among academic medical researchers. METHODS: From February to March 2020, an anonymous survey was conducted among academic medical researchers, pharmaceutical industry professionals, and IP researchers based in Japan. Overall response rate was 27.4% (192/700). We conducted an analysis of variance for H1 and used the PLS-SEM model for H2 in order to verify the hypotheses. RESULTS: The results confirmed that the mean calculated from the responses of the academic medical researchers was significantly lower than the mean of pharmaceutical company personnel when responses to patenting an emerging technology or drug for the advancement of medicine were analyzed. In addition, we found that a causal relationship between academic medical researchers' understanding of patents and their positive impressions on patents, depending on the degree to which they consider that the patent system is to encourage and promote new inventions. CONCLUSION: We conclude that a contrasting perception of patents not only exists between academic medical researchers and pharmaceutical company personnel but also it is caused by their apprehension of patents. More efforts to promote the raison d'être of the patent system among academic medical researchers will enable them to view pharmaceutical patents in a more positive light. Through this study, the pertinence to promote academic drug discoveries has been uncovered.

Measurement and differentiation of banana juice scent using an electronic nose FF-2A.

BACKGROUND: Banana juice is becoming a popular beverage in Japan and the number of soft-drink stands or shops that take great care and pride in the quality of their products has been increasing. This study aims to measure the scent of banana juice from different brands using the electronic (e-) nose FF-2A in order to identify the characteristics, time-related changes, and the differences among them. METHODS: We standardized the scent value of banana juice measured using FF-2A and determined the absolute value in three different shops. We compared the similarities in samples from each shop with axis data created using standardized measurement. With FF-2A we identified the scent common to all banana juice samples from the composite scent and numerically showed the similarity to the reference gas. RESULTS: The juices from each shop had their own characteristics and we were able to identify the difference between some of these. The response of FF-2A varied according to the increase/decrease in the number of characteristic molecules measured by GC-MS such as overtime fluctuations in the gas. These data were shown along with the differences between the various banana juices. CONCLUSIONS: FF-2A was able to identify the scent of banana juice at each banana shop as well as time-related changes. By combining GC-MS, we were able to evaluate scent components that changed over time. The results using the electronic nose may prove useful for objective evaluation and comparison of scent with other types of juices.

Effect of Phosphoribosyltransferase Down-regulation on Malignant Glioma Cell Characteristics.

BACKGROUND/AIM: Nicotinamide phosphoribosyl-transferase (NAMPT) is a rate-limiting enzyme in the pathway synthesizing nicotinamide adenine dinucleotide (NAD (+)) from nicotinamide (NAM). Glioma tissues exhibit up-regulated NAMPT expression associated with a poor prognosis of patients. To determine if NAMPT can be a molecular therapeutic target, we investigated the effects of short hairpin RNA (shRNA)-mediated NAMPT down-regulation. MATERIALS AND METHODS: We designed shRNA to NAMPT and transfected to T98G cells. The characteristics of these cells were analyzed. RESULTS: The NAMPT shRNA-transfected cells exhibited delayed cell growth. However, there was no difference in the increase of sensitivity to temozolomide (TMZ) or X-ray irradiation between the NAMPT and scramble shRNA-transfected cells. The expression of NAMPT in the NAMPT shRNA-transfected cells increased with cell passage. Additionally, the shRNA-mediated transfection was associated with enhanced expression of quinolinic acid phosphoribo-syltransferase (QPRT). CONCLUSION: shRNA-mediated NAMPT down-regulation may not decrease the NADt to a sufficient level to increase TMZ/radiation sensitivity.

Osteogenic cocktail induces calcifications in human breast cancer cell line via placental alkaline phosphatase expression.

Breast cancer is frequently characterized by calcifications in mammography. The mechanism for calcifications in breast cancer is not completely known. Understanding this mechanism will improve diagnostic accuracy. Herein, we demonstrated that calcifications occur and that alkaline phosphatase enzyme activity increases in MDA-MB-231 cells cultured using an osteogenic cocktail-containing medium. Microarray transcript analysis showed that the PI3K-Akt signaling pathway was significantly involved, with recruitment of placental alkaline phosphatase. Calcifications and alkaline phosphatase enzyme activity were suppressed by silencing placental alkaline phosphatase using a small interfering RNA. Inhibition of the PI3K-Akt signaling pathway suppressed phospho-c-Jun and placental alkaline phosphatase and resulted in absence of calcifications. These findings reveal that breast cancer cells acquire alkaline phosphatase enzyme activity via placental alkaline phosphatase expression and suggest that breast calcification formation is closely associated with the PI3K-Akt signaling pathway.

Involvement of Corticotropin-releasing Hormone-related Peptides in Cellular Stress Caused by Anticancer Drugs in Colorectal Cancer.

BACKGROUND/AIM: The expression of corticotropin-releasing hormone (CRH)-related peptides involved in stress response in colorectal cancer has been reported. We examined the involvement of CRH-related peptides in cellular stress caused by anticancer drugs in colorectal cancer. MATERIALS AND METHODS: Changes in the expression levels of CRH-related peptides and their receptors in HCT116, DLD-1, and SW480 cell lines after fluorouracil (5-FU) loading were evaluated. Effects of the receptor antagonist against DNA synthesis disorder caused by 5-FU and SN-38 were evaluated using the 3H-labeled deoxyribonucleoside incorporation assay. RESULTS: No changes in the mRNA expression levels of CRH-related peptides (UCN and UCN2) -and their receptors (CRHR1 and CRHR2) were observed. Addition of antagonists to cells with DNA synthesis disorder caused by 5-FU and SN-38 showed no differences in the incorporation of 3H-labeled deoxyribonucleoside. CONCLUSION: CRH-related peptides showed no effect on the stress response to anticancer drugs nor on DNA synthesis disorder in colorectal cancer.

MicroRNA-200b and -301 are associated with gemcitabine response as biomarkers in pancreatic carcinoma cells.

Chemotherapy resistance (congenital or acquired) is one of the principal challenges for the treatment of pancreatic carcinoma. Recent evidence has demonstrated that epithelial to mesenchymal transition (EMT) is associated with chemoresistance in pancreatic carcinoma cells. However, the molecular mechanism underlying the development of chemoresistance remains unknown, and limited therapeutic options are available. Therefore, to anticipate individual chemosensitivity or acquired chemoresistance for patients with pancreatic carcinoma, predictive biomarkers are urgently required. Extensive evidence suggests that microRNAs (miRNAs) serve a crucial role in regulating EMT. The aim of this study was to examine the potential role of miRNA (miR)­200b and miR­301 in predicting the chemo­responses to treatment for pancreatic carcinoma. The present results demonstrate that miR­200b expression predicted chemo­sensitivity and may have potential as a biomarker. In six different pancreatic carcinoma cell lines (Capan­1, Capan­2, Panc­1, MIAPaCa­2, BxPC­3 and PL45 cells), the expression of miR­200b correlated positively with chemosensitivity. Moreover, the enhanced expression of miR­200b increased chemosensitivity and induced mesenchymal to epithelial transition. Conversely, miR­301 modulated gemcitabine resistance and induced EMT through the downregulation of cadherin 1 expression. In addition, gemcitabine­resistant cells (Capan­2 and Panc­1) exhibited upregulated miR­301 expression and downregulated gemcitabine­induced apoptosis. In summary, these two miRNAs may serve roles as biomarkers in pancreatic carcinoma, miR­200b expression may predict chemosensitivity, and elevated miR­301 expression may have potential applications in the prediction of acquired gemcitabine resistance.

Effect of Heparin-Protamine Treatment on Thyroid Cancer Cell Lines.

BACKGROUND: Anticoagulation therapy is often used to prevent stroke in patients with thyroid cancer. However, the effects of heparin and protamine on thyroid cancer are unknown; therefore, we examined the response of thyroid cancer cell lines to heparin and protamine. MATERIALS AND METHODS: Cytotoxic assay for heparin-protamine treatment was examined on SW1736, 8505c and 8305c cell lines. RESULTS: The half-maximal inhibitory concentration of the heparin-protamine treatment was 82.1 µg/ml for the SW1736 cell line, 10.4 µg/ml for 8505c, and 17.8 µg/ml for 8305c. Each cell line expresses fibronectin, with SW1736 expressing mutant fibronectin; thus, it is possible that mutant fibronectin may prevent the antitumor effect in SW1736 cells. CONCLUSION: The SW1736 cell line did not show any antitumor effect after heparin-protamine treatment. Further research is needed on why heparin-protamine treatment does not exert an antitumor effect on SW1736 cells, and the results of this research could mean that the heparin-protamine treatment might be used to provide antitumor effects in some thyroid cancer cases.

Temozolomide does not influence the transcription or activity of matrix metalloproteinases 9 and 2 in glioma cell lines.

Glioblastoma multiforme (GBM) is a treatment-resistant malignancy with poor prognosis. Temozolomide (TMZ) is widely used as a first-line drug for GBM. Although this improves patient prognosis, it does not completely eradicate the tumour. Even after total surgical resection, GBM can exhibit uncontrollable invasiveness at the tumour margins owing to activation of matrix metalloproteinases (MMPs) such as MMP-2 and -9; these degrade collagen IV in the basement membrane, which normally prevents cancer invasion. TMZ induces DNA damage and activates transcription factors including c-jun, c-fos, nuclear factor-κß, and early growth response protein-1, which have putative binding sites on the MMP-9 promoter. TMZ may therefore enhance tumour invasion by stimulating MMP-9 transcription and enzymatic activity. To test this hypothesis, we investigated MMP-2 and -9 mRNA transcription and activity in GBM cell lines treated with TMZ. Human A172 GBM cells were exposed to TMZ (25% and 50% inhibitory concentrations) for 24 or 48h; cell cycle distribution and mRNA levels of MMP-2 and -9 were evaluated using flow cytometry and semi-quantitative reverse transcription PCR, respectively. MMP-2 and -9 enzymatic activities were assessed using gelatin zymography in human A172 and U373 MG GBM cells exposed to TMZ under the same conditions. TMZ altered A172 cell cycle distribution, but not MMP-2 or -9 mRNA levels. TMZ did not affect MMP-2 or -9 enzymatic activities in A172 or U373 MG cells. These findings indicated that TMZ is therefore unlikely to promote GBM invasiveness.

The Powdering Process with a Set of Ceramic Mills for Green Tea Promoted Catechin Extraction and the ROS Inhibition Effect.

For serving green tea, there are two prominent methods: steeping the leaf or the powdered leaf (matcha style) in hot water. The purpose of the present study was to reveal chemical and functional differences before and after the powdering process of green tea leaf, since powdered green tea may contribute to expanding the functionality because of the different ingesting style. In this study, we revealed that the powdering process with a ceramic mill and stirring in hot water increased the average extracted concentration of epigallocatechin gallate (EGCG) by more than three times compared with that in leaf tea using high-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass Spectrometry (LC-MS/MS) analyses. Moreover, powdered green tea has a higher inhibition effect of reactive oxygen species (ROS) production in vitro compared with the same amount of leaf tea. Our data suggest that powdered green tea might have a different function from leaf tea due to the higher catechin contents and particles.

BRAF V600E-mutated diffuse glioma in an adult patient: a case report and review.

Recent advances in genomic technology and genome-wide analysis have identified key molecular alterations that are relevant to the diagnosis and prognosis of brain tumors. Molecular information such as mutations in isocitrate dehydrogenase (IDH) genes or 1p/19q co-deletion status will be more actively incorporated into the histological classification of diffuse gliomas. BRAF V600E mutations are found frequently in circumscribed low-grade gliomas such as pleomorphic xanthoastrocytoma (PXA) and extra-cerebellar pilocytic astrocytoma, or epithelioid glioblastomas (E-GBM), a rare variant of GBM. This mutation is relatively rare in other types of diffuse gliomas, especially in adult onset cases. Here, we present an adult onset case of IDH wild-type/BRAF V600E-mutated diffuse glioma, evolving from grade III to grade IV. The tumor displayed atypical exophytic growth and had unusual histological features not fully compatible with, but indicative of PXA and E-GBM. We discuss differential diagnosis of the tumor, and review previously described diffuse gliomas with the BRAF V600E mutation.

Cell Membrane Perforation with Photosensitizer and a Brush-shaped Soft-polymer Sheet Using a Malignant Glioma Cell Line.

BACKGROUND: Transduction of foreign molecules into cells is an important technique to investigate the functions of corresponding molecules and/or targets. Recently, a mass-producible nanoprinting perforator was devised enabling for large-scale, high-performance drug or nucleic-acid transfer into cells without cell damage. Since little is known on the performance of the system, we investigated its effects on a malignant glioma cell line. MATERIALS AND METHODS: Photosensitization was performed by the Cell Stamper CP-01. The malignant U373MG glioma cell line was used for transduction. RESULTS: Photosensitization transduced FITC-conjugated albumin into cells. Trypan blue inclusion test demonstrated membrane disintegration by the procedure and scanning electron microscopy disclosed perforation of the cell membrane. CONCLUSION: Local oxidation reaction during the nanoprinting caused reversible membrane perforation. Morphological findings from the current study support the above mechanism, therefore the specific printing system might be convenient for transduction of foreign molecules into malignant glioma cells.