Establishment of tumor microenvironment-preserving organoid model from patients with intracranial meningioma.

BACKGROUND: Although meningioma is the most common primary brain tumor, treatments rely on surgery and radiotherapy, and recurrent meningiomas have no standard therapeutic options due to a lack of clinically relevant research models. Current meningioma cell lines or organoids cannot reflect biological features of patient tumors since they undergo transformation along culture and consist of only tumor cells without microenvironment. We aim to establish patient-derived meningioma organoids (MNOs) preserving diverse cell types representative of the tumor microenvironment. METHODS: The biological features of MNOs were evaluated using WST, LDH, and collagen-based 3D invasion assays. Cellular identities in MNOs were confirmed by immunohistochemistry (IHC). Genetic alteration profiles of MNOs and their corresponding parental tumors were obtained by whole-exome sequencing. RESULTS: MNOs were established from four patients with meningioma (two grade 1 and two grade 2) at a 100% succession rate. Exclusion of enzymatic dissociation-reaggregation steps endowed MNOs with original histology and tumor microenvironment. In addition, we used a liquid media culture system instead of embedding samples into Matrigel, resulting in an easy-to-handle, cost-efficient, and time-saving system. MNOs maintained their functionality and morphology after long-term culture (> 9 wk) and repeated cryopreserving-recovery cycles. The similarities between MNOs and their corresponding parental tumors were confirmed by both IHC and whole-exome sequencing. As a representative application, we utilized MNOs in drug screening, and mifepristone, an antagonist of progesterone receptor, showed prominent antitumor efficacy with respect to viability, invasiveness, and protein expression. CONCLUSION: Taken together, our MNO model overcame limitations of previous meningioma models and showed superior resemblance to parental tumors. Thus, our model could facilitate translational research identifying and selecting drugs for meningioma in the era of precision medicine.

Intraocular Lens Unfurling Time Exponentially Decays with Increased Solution Temperature.

Purpose: Intraocular lens (IOL) unfurling can be a rate-limiting step in cataract surgery, limiting operative efficiency. Furthermore, inefficient unfurling has important implications for clinical outcomes. We examine the effects of solution temperature on IOL unfurling time using three in vitro models of the ocular environment. Methods: IOLs were injected into a 6-well plate filled with balanced salt solution (BSS), dispersive ophthalmic viscoelastic device (OVD), or cohesive OVD. Experiments were also performed in a plastic eye filled with dispersive or cohesive OVD. IOL unfurling time was recorded against the temperature of the respective solution. Results: IOL unfurling time decayed exponentially as solution temperature increased in all experiments, including the BSS-filled 6-well plate, the OVD-filled 6-well plate, and the OVD-filled plastic eye. IOLs failed to unfurl within 10 min at 10°C, below the glass transition temperature of the tested IOLs. Increasing solution temperature from 20°C to 30°C decreases IOL unfurling by greater than 2 min. Further heating to 40°C did not significantly decrease IOL unfurling time. Conclusion: Increased solution temperature rapidly decreases IOL unfurling time in vitro. IOLs do not unfurl within a clinically acceptable timeframe at or below their glass transition temperature. Increased BSS and/or OVD temperature may be a potential method to decrease IOL unfurling time in cataract surgery. However, future research is needed to elucidate potential consequences of warmed BSS and/or OVD on post-operative outcomes. This study demonstrates the potential for temperature regulation to decrease cataract surgery operative time and provides preliminary evidence to justify future clinical validation of this relationship.

During cataract surgery, a prosthetic intraocular lens (IOL) is inserted into the eye once the clouded lens is removed. The IOL must then unfurl before the procedure can proceed. When IOLs fail to unfurl or unfurl slowly, this can delay the operation and may even cause post-operative complications. Thus, we studied the effect temperature may have on IOL unfurling time to optimize this segment of the operation. We injected IOLs into solutions of saline (balanced salt solution) or ophthalmic viscoelastic device (OVD), two fluids injected into the eye during surgery. In both a well plate and a plastic eye, we found that increasing the temperature of the solution significantly affected IOL unfurling time. Specifically, heating the solution from refrigeration to room temperature decreased unfurling time from over 10 min to less than four. Heating to physiological temperature further decreased unfurling time to less than a minute. Our results show promise for potentially utilizing heated BSS and/or OVD to accelerate IOL unfurling and decrease cataract surgery operative time.

Suppression of Metastatic Ovarian Cancer Cells by Bepridil, a Calcium Channel Blocker.

Although surgery followed by platinum-based therapy is effective as a standard treatment in the early stages of ovarian cancer, the majority of cases are diagnosed at advanced stages, leading to poor prognosis. Thus, the identification of novel therapeutic drugs is needed. In this study, we assessed the effectiveness of bepridil-a calcium channel blocker-in ovarian cancer cells using two cell lines: SKOV-3, and SKOV-3-13 (a highly metastatic clone of SKOV-3). Treatment of these cell lines with bepridil significantly reduced cell viability, migration, and invasion. Notably, SKOV-3-13 was more sensitive to bepridil than SKOV-3. The TGF-ß1-induced epithelial-mesenchymal transition (EMT)-like phenotype was reversed by treatment with bepridil in both cell lines. Consistently, expression levels of EMT-related markers, including vimentin, ß-catenin, and Snail, were also substantially decreased by the treatment with bepridil. An in vivo mouse xenograft model was used to confirm these findings. Tumor growth was significantly reduced by bepridil treatment in SKOV-3-13-inoculated mice, and immunohistochemistry showed consistently decreased expression of EMT-related markers. Our findings are the first to report anticancer effects of bepridil in ovarian cancer, and they suggest that bepridil holds significant promise as an effective therapeutic agent for targeting metastatic ovarian cancer.

Proteolytic pan-RAS Cleavage Leads to Tumor Regression in Patient-derived Pancreatic Cancer Xenografts.

The lack of effective RAS inhibition represents a major unmet medical need in the treatment of pancreatic ductal adenocarcinoma (PDAC). Here, we investigate the anticancer activity of RRSP-DTB, an engineered biologic that cleaves the Switch I of all RAS isoforms, in KRAS-mutant PDAC cell lines and patient-derived xenografts (PDX). We first demonstrate that RRSP-DTB effectively engages RAS and impacts downstream ERK signaling in multiple KRAS-mutant PDAC cell lines inhibiting cell proliferation at picomolar concentrations. We next tested RRSP-DTB in immunodeficient mice bearing KRAS-mutant PDAC PDXs. Treatment with RRSP-DTB led to ≥95% tumor regression after 29 days. Residual tumors exhibited disrupted tissue architecture, increased fibrosis and fewer proliferating cells compared with controls. Intratumoral levels of phospho-ERK were also significantly lower, indicating in vivo target engagement. Importantly, tumors that started to regrow without RRSP-DTB shrank when treatment resumed, demonstrating resistance to RRSP-DTB had not developed. Tracking persistence of the toxin activity following intraperitoneal injection showed that RRSP-DTB is active in sera from immunocompetent mice for at least 1 hour, but absent after 16 hours, justifying use of daily dosing. Overall, we report that RRSP-DTB strongly regresses hard-to-treat KRAS-mutant PDX models of pancreatic cancer, warranting further development of this pan-RAS biologic for the management of RAS-addicted tumors.

Multi-Stage Temporal Convolutional Network with Moment Loss and Positional Encoding for Surgical Phase Recognition.

In recent times, many studies concerning surgical video analysis are being conducted due to its growing importance in many medical applications. In particular, it is very important to be able to recognize the current surgical phase because the phase information can be utilized in various ways both during and after surgery. This paper proposes an efficient phase recognition network, called MomentNet, for cholecystectomy endoscopic videos. Unlike LSTM-based network, MomentNet is based on a multi-stage temporal convolutional network. Besides, to improve the phase prediction accuracy, the proposed method adopts a new loss function to supplement the general cross entropy loss function. The new loss function significantly improves the performance of the phase recognition network by constraining un-desirable phase transition and preventing over-segmentation. In addition, MomnetNet effectively applies positional encoding techniques, which are commonly applied in transformer architectures, to the multi-stage temporal convolution network. By using the positional encoding techniques, MomentNet can provide important temporal context, resulting in higher phase prediction accuracy. Furthermore, the MomentNet applies label smoothing technique to suppress overfitting and replaces the backbone network for feature extraction to further improve the network performance. As a result, the MomentNet achieves 92.31% accuracy in the phase recognition task with the Cholec80 dataset, which is 4.55% higher than that of the baseline architecture.

Optimal Postoperative Surveillance Strategies for Colorectal Cancer: A Retrospective Observational Study.

This study aimed to assess whether surveillance intensity is associated with recurrence and survival in colorectal cancer (CRC) patients. Overall, 3794 patients with pathologic stage I-III CRC who underwent radical surgery between January 2012 and December 2014 were examined. Surveillance comprised abdominopelvic computed tomography (CT) every 6 months and chest CT annually for 5 years. Patients who underwent more than and less than an average of three imaging examinations annually were assigned to the high-intensity (HI) and low-intensity (LI) groups, respectively. Demographics were similar in both groups. T and N stages were higher and perineural and lymphovascular invasion were more frequent in the HI group (p < 0.001 each). The mean overall survival (OS) was similar for both groups; however, recurrence-free survival (RFS) was longer (p < 0.001) and post-recurrence survival (PRS) was shorter (p = 0.024) in the LI group. In the multivariate analysis, surveillance intensity was associated with RFS (p < 0.001) in contrast to PRS (p = 0.731). In patients with high recurrence risk predicted using the nomogram, OS was longer in the HI group (p < 0.001). A higher imaging frequency in patients at high risk of recurrence could be expected to lead to a slight increase in PRS but does not improve OS. Therefore, rather than increasing the number of CT scans in high-risk patients, other imaging modalities or innovative approaches, such as liquid biopsy, are required.

Antibacterial Activity of Caffeic Acid Combined with UV-A Light against Escherichia coli O157:H7, Salmonella enterica Serovar Typhimurium, and Listeria monocytogenes.

The aim of this study was to evaluate the antibacterial activity of caffeic acid (CA), which is a natural polyphenol, combined with UV-A light against the representative foodborne bacteria Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes. Data regarding the inactivation of these bacteria and its dependence on CA concentration, light wavelength, and light dose were obtained. E. coli O157:H7 and Salmonella Typhimurium were reduced to the detection limit when treated with 3 mM CA and UV-A for 3 J/cm2 and 4 J/cm2, respectively, and 5 J/cm2 treatment induced 3.10 log reduction in L. monocytogenes. To investigate the mechanism for inactivation of Salmonella Typhimurium and L. monocytogenes, measurement of polyphenol uptake, membrane damage assessment, enzymatic activity assay, and transmission electron microscopy (TEM) were conducted. It was revealed that CA was significantly (P < 0.05) absorbed by bacterial cells, and UV-A light allowed a higher uptake of CA for both pathogens. Additionally, CA plus UV-A treatment induced significant (P < 0.05) cell membrane damage. In the enzymatic activity assay, the activities of both pathogens were reduced by CA, and a greater reduction occurred by use of CA plus UV-A. Moreover, transmission electron microscopy (TEM) images indicated that CA plus UV-A treatment notably destroyed the intercellular structure. In addition, antibacterial activity was also observed in commercial apple juice, which showed results similar to those obtained from phosphate-buffered saline (PBS), resulting in a significant (P < 0.05) reduction for all three pathogens without any changes in color parameters (L*, a*, and b*), total phenolic compounds, and DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging activity. IMPORTANCE Photodynamic inactivation (PDI), which involves photoactivation of a photosensitizer (PS), is an emerging field of study, as it effectively reduces various kinds of microorganisms. Although there are several PSs that have been used for PDI, there is a need to find naturally occurring PSs for safer application in the food industry. Caffeic acid, a natural polyphenol found in most fruits and vegetables, has recently been studied for its potential to act as a novel photosensitizer. However, no studies have been conducted regarding its antibacterial activity depending on treatment conditions and its antibacterial mechanism. In this study, we closely examined the effectiveness of caffeic acid in combination with UV-A light for inactivating representative foodborne bacteria in liquid medium. Therefore, the results of this research are expected to be utilized as basic data for future application of caffeic acid in PDI, especially when controlling pathogens in liquid food processing.

Non-Ablative 1927 nm Fractional Thulium Fiber Laser: New, Promising Treatment Modality for Riehl's Melanosis.

BACKGROUND AND OBJECTIVES: The treatment of Riehl's melanosis, also known as pigmented contact dermatitis, is highly challenging. Intense pulsed light (IPL) and 1064 nm Q-switched Nd:Yag (QS-Nd:YAG) laser are reported to have some efficacy. However, no single effective treatment has yet been identified. In this study, we demonstrated the efficacy and safety of the non-ablative 1927 nm fractional thulium fiber laser (TFL, LASEMD™; Lutronic Corp., Goyang, Korea) for patients with Riehl's melanosis. STUDY DESIGN/MATERIALS AND METHODS: A retrospective chart and photographic review of nine patients with Riehl's melanosis, who had received at least three sessions of TFL treatment, was performed. Before the start of TFL treatment, combination treatment with a topical cream containing hydroquinone, low-fluence QS-Nd:YAG laser, pulsed dye laser, and IPL was used with variable and discouraging effects. Seven patients were treated on the face and two patients on the neck with three to seven sessions at 1-month intervals. Clinical improvement was assessed using clinical photos taken before and after every treatment session according to dermal pigmentation area and severity index (DPASI) and a quartile grading scale by two blinded dermatologists. RESULTS: Patients underwent three to seven sessions of TFL treatment depending on severity of pigmentation. Of nine patients, six demonstrated a clinical improvement of 51%-75%, one demonstrated an improvement of 76%-100%, and two showed an improvement of 26%-50% after treatment. The DPASI was significantly decreased from 9.55 to 5.25 on average. Melanin index was decreased after treatment in two patients whose melanin index were measured at initial visits. Treatment-related adverse events, such as scarring or postinflammatory hyperpigmentation (PIH), were not observed in all patients except for transient erythema and swelling. CONCLUSIONS: This report suggests that TFL could be an alternative and/or additive treatment option for hyperpigmentation in intractable Riehl's melanosis and might be a promising treatment for PIH caused by any reason including Riehl's melanosis. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.

A Prospective, Split-Face, Comparative Study of Combined Treatment With Fractional Microneedle Radiofrequency and Nonablative 1927-nm Fractional Thulium Fiber Laser for Wrinkle Treatment.

BACKGROUND: Fractional microneedle radiofrequency (FMR) and nonablative 1927-nm fractional thulium fiber laser (TFL) are widely used for skin rejuvenation treatment. OBJECTIVES: To investigate the efficacy and safety of combined treatment with both devices for wrinkles. PATIENTS AND METHODS: Twenty-five patients with wrinkles were enrolled. One side of the face was treated with FMR alone, while the other side was treated with a combination of FMR and TFL. Each treatment consisted of 3 sessions at four-week intervals and patients were followed up 12 weeks after the last treatment. Overall improvement was assessed by patient global assessment (PGA) and investigator global assessment (IGA). Depression scores for the evaluation of wrinkles were objectively assessed by Antera 3D system. RESULTS: Both sides of the face led to clinical improvement in both mean PGA and IGA. Combination treatment demonstrated a greater improvement in both mean PGA and IGA compared with FMR alone. In addition, wrinkle grading scales and depression scores showed greater improvement in the combination group than in FMR alone. CONCLUSION: This study demonstrated that FMR and TFL comprise a good combination treatment for the treatment of wrinkles because both treatments have a synergistic effect on wrinkle improvement.

Sirolimus and MMF are insufficient immunosuppressants for regulation of the proliferation of CD133+EpCAM+ cell populations in HCC cell lines.

Studies on effective immunosuppressive strategies for the management of patients undergoing a liver transplantation (LT) due to hepatocellular carcinoma (HCC) are limited. In the present study, immunosuppressive candidates predicted to exhibit beneficial immunosuppressive and tumor-suppressive effects in patients with HCC were assessed using Huh7 and HEP3B HCC cells, which have high proportions of CD133+EpCAM+ cancer stem cell (CSC) populations. The immunosuppressants assessed were sirolimus, tacrolimus, cyclosporine A and mycophenolate mofetil (MMF), and their activities were assessed on CSCs. Sirolimus and MMF reduced the proliferation of Huh7 and HEP3B cells; however, the proportion of CD133+EpCAM+ was notably increased in treated Huh7 cells. Sirolimus treatment alone resulted in G0-G1 cell cycle arrest at all doses in all Huh7 and CD133-EpCAM- populations; however, CD133+EpCAM+ populations showed only slight G1 arrest at higher doses only. In contrast, S-phase arrest was induced at all doses in the Huh7, CD133-EpCAM- and CD133+EpCAM+ populations by MMF. Sirolimus and MMF effectively reduced the proliferation of Huh7 and HEP3B cells, but did not exert a notable effect on the CD133+EpCAM+ cells. Therefore, therapeutic strategies utilizing Sirolimus and MMF should be further studied in vivo for regulation of CSC populations in order to reduce HCC recurrence rates.

An engineered chimeric toxin that cleaves activated mutant and wild-type RAS inhibits tumor growth.

Despite nearly four decades of effort, broad inhibition of oncogenic RAS using small-molecule approaches has proven to be a major challenge. Here we describe the development of a pan-RAS biologic inhibitor composed of the RAS-RAP1-specific endopeptidase fused to the protein delivery machinery of diphtheria toxin. We show that this engineered chimeric toxin irreversibly cleaves and inactivates intracellular RAS at low picomolar concentrations terminating downstream signaling in receptor-bearing cells. Furthermore, we demonstrate in vivo target engagement and reduction of tumor burden in three mouse xenograft models driven by either wild-type or mutant RAS Intracellular delivery of a potent anti-RAS biologic through a receptor-mediated mechanism represents a promising approach to developing RAS therapeutics against a broad array of cancers.

ATP-P2X7-Induced Inflammasome Activation Contributes to Melanocyte Death and CD8+ T-Cell Trafficking to the Skin in Vitiligo.

Extracellular adenosine 5'-triphosphate (ATP) is a well-known inflammasome-activating signal. Emerging evidence demonstrates a critical role for inflammasome activation in vitiligo pathogenesis. However, the specific molecular mechanism of inflammasome-dependent melanocyte degeneration in vitiligo is still not clear. This study presents how extracellular ATP, released from keratinocytes by oxidative stress, affects melanocyte survival in vitiligo skin. H2O2-induced oxidative injury increased ATP release from keratinocytes and skin tissues. The high concentration of extracellular ATP induced both ROS production and cell death in melanocytes. Treatment with ATP caused the activation of caspase-1 as well as the production of active forms of IL-1ß and IL-18 via P2X7 receptor in keratinocytes and melanocytes. Lesional and perilesional skin of vitiligo showed higher levels of ATP as well as upregulation of active caspase-1 compared with nonlesional skin, suggesting its possible role in inflammasome activation in vitiligo. Moreover, the elevated expression of CXCL9 in keratinocytes, mediated through ATP/P2X7 receptor-dependent inflammasome activation, was responsible for CLA+CD8+ T-cell chemotaxis into the skin. These results demonstrate that extracellular ATP as a danger signal activates the inflammasome pathway and increases cutaneous chemotaxis of CD8+ T cells via CXCL9 in vitiligo. Therefore, targeting ATP-P2X7 signaling may be a potential strategy for vitiligo treatment.

Postthyroidectomy obesity in a Korean population: does the extent of surgery matter?

PURPOSE: The purpose of this study was to investigate the prevalence of postthyroidectomy obesity, and the relationship between the extent of thyroidectomy and obesity. METHODS: A survey conducted at an outpatient clinic from June to October 2014 and retrospective charts for patients undergoing thyroidectomy at Konkuk University Medical Centers from June 2009 to December 2013 were reviewed. We compared clinical characteristics and pre- and postoperative obesity-related factors in 227 patients who underwent total thyroidectomy or lobectomy. RESULTS: Patients included 39 males and 188 females with a mean age of 46.0 ± 11.0 years; the mean follow-up period was 23.9 ± 16.7 months, and 90 of the 227 patients showed postthyroidectomy obesity. In effect of operative extent on postoperative obesity, patients who underwent TT (48.2 years) than those who underwent lobectomy (43.4 years). TT group had longer follow-up and the frequency of menopause was higher than in the lobectomy group. No differences in postthyroidectomy obesity, body weight change, or body mass index (BMI), change among 2 groups. The predictors of postthyroidectomy obesity were older age, female, heavy alcohol consumption (P = 0.029), higher preoperative BMI (P < 0.001), larger postoperative weight gain (P = 0.024), and larger BMI change. However, the extent of thyroidectomy did not affect postthyroidectomy obesity. Preoperative BMI (P < 0.001) and heavy alcohol consumption (P = 0.03) were independent factors of postthyroidectomy obesity. CONCLUSION: The extent of thyroidectomy does not affect postthyroidectomy obesity. Preoperative BMI and heavy alcohol consumption are risk factors for postthyroidectomy obesity. Studies are needed to suggest preoperative life style modification to prevent postthyroidectomy obesity.

Effect of PNPLA3 I148M polymorphism on histologically proven non-alcoholic fatty liver disease in liver transplant recipients.

AIM: PNPLA3 I148M polymorphism (rs738409 C>G) is the most important and best-known polymorphism for non-alcoholic fatty liver disease (NAFLD). However, little is known about the effect of this polymorphism on NAFLD after liver transplantation (LT). We aimed to evaluate the association between this polymorphism and post-LT NAFLD. METHODS: We designed a prospective case-control study. Among adult recipients who underwent LT between April 2014 and October 2015, those whose whole blood was preoperatively collected for genotyping in both recipients and coupled donors and those who underwent protocol biopsy at 1 year post-LT were enrolled. RESULTS: A total of 32 recipients were enrolled. Histologically proven steatosis (≥5%) was present in 28.1% of patients at a mean time of 12.7 ± 2.0 months after LT. Moderate and more severe steatosis (≥33%) was present in 9.4%. One year after LT, steatosis was present in 50.0% of homozygous recipients with the rs738409-G allele. It was present in 27.3% of heterozygous recipients with the rs738409-G allele, and in 9.1% (P = 0.041) of recipients with rs738409-CC. The genotype of the donor was not significantly (P = 0.647) associated with post-LT NAFLD. When both recipient and coupled donor showed heterogeneous or homozygous genotype of the rs738409-G allele, there was significantly more post-LT NAFLD compared to that in others (47.1% vs. 6.7%; P = 0.018). In univariate and multivariate analyses, only the presence of the rs738409-G risk allele in both donor and recipient was a significant risk factor for post-LT NALFD (relative risk, 26.95; P = 0.048). CONCLUSIONS: PNPLA3 I148M polymorphism can significantly affect histologically proven NAFLD at 1 year post-LT.

Genetically encoded photocrosslinkers locate the high-affinity binding site of antidepressant drugs in the human serotonin transporter.

Despite the well-established role of the human serotonin transporter (hSERT) in the treatment of depression, the molecular details of antidepressant drug binding are still not fully understood. Here we utilize amber codon suppression in a membrane-bound transporter protein to encode photocrosslinking unnatural amino acids (UAAs) into 75 different positions in hSERT. UAAs are incorporated with high specificity, and functionally active transporters have similar transport properties and pharmacological profiles compared with wild-type transporters. We employ ultraviolet-induced crosslinking with p-azido-L-phenylalanine (azF) at selected positions in hSERT to map the binding site of imipramine, a prototypical tricyclic antidepressant, and vortioxetine, a novel multimodal antidepressant. We find that the two antidepressants crosslink with azF incorporated at different positions within the central substrate-binding site of hSERT, while no crosslinking is observed at the vestibular-binding site. Taken together, our data provide direct evidence for defining the high-affinity antidepressant binding site in hSERT.

Smoking Cessation Failure Among Korean Adolescents.

The aim of this study was to identify smoking cessation failure subgroups among Korean adolescents. Participants were 379 smoking adolescents who joined a smoking cessation program. A questionnaire and a cotinine urine test were administered before the program began. Three months after the program ended, the cotinine urine test was repeated. A decision-tree model identified seven subgroups with low or high smoking cessation rates. The predictors of smoking cessation were intention to stop smoking, initiation of smoking, amount of cigarette use, self-efficacy, and paternal smoking status. The subgroup with the lowest smoking cessation rate included adolescents who did not have any intention to stop smoking and who had started smoking after eighth grade, and none of the participants in this group stopped smoking. The results of this study provide crucial information for tailored smoking cessation programs.