Increased Multiplexity in Optical Tissue Clearing-Based Three-Dimensional Immunofluorescence Microscopy of the Tumor Microenvironment by Light-Emitting Diode Photobleaching.

Optical tissue clearing and three-dimensional (3D) immunofluorescence (IF) microscopy is transforming imaging of the complex tumor microenvironment (TME). However, current 3D IF microscopy has restricted multiplexity; only 3 or 4 cellular and noncellular TME components can be localized in cleared tumor tissue. Here we report a light-emitting diode (LED) photobleaching method and its application for 3D multiplexed optical mapping of the TME. We built a high-power LED light irradiation device and temperature-controlled chamber for completely bleaching fluorescent signals throughout optically cleared tumor tissues without compromise of tissue and protein antigen integrity. With newly developed tissue mounting and selected region-tracking methods, we established a cyclic workflow involving IF staining, tissue clearing, 3D confocal microscopy, and LED photobleaching. By registering microscope channel images generated through 3 work cycles, we produced 8-plex image data from individual 400 µm-thick tumor macrosections that visualize various vascular, immune, and cancer cells in the same TME at tissue-wide and cellular levels in 3D. Our method was also validated for quantitative 3D spatial analysis of cellular remodeling in the TME after immunotherapy. These results demonstrate that our LED photobleaching system and its workflow offer a novel approach to increase the multiplexing power of 3D IF microscopy for studying tumor heterogeneity and response to therapy.

Pembrolizumab versus paclitaxel for previously treated advanced gastric or gastroesophageal junction cancer (KEYNOTE-063): A randomized, open-label, phase 3 trial in Asian patients.

BACKGROUND: KEYNOTE-063 (NCT03019588) investigated pembrolizumab versus paclitaxel as second-line therapy in Asian patients with advanced programmed death ligand 1 (PD-L1)-positive (combined positive score ≥1) gastric/gastroesophageal junction (GEJ) cancer. METHODS: This randomized, open-label, phase 3 study was conducted at 36 medical centers in China (mainland), Malaysia, South Korea, and Taiwan. Patients were randomly assigned 1:1 to 200 mg of pembrolizumab intravenously every 3 weeks for ≤2 years or 80 mg/m2 of paclitaxel intravenously every week. Primary end points were overall survival (OS) and progression-free survival (PFS). Secondary end points were objective response rate (ORR) per Response Evaluation Criteria in Solid Tumors version 1.1 and safety. RESULTS: Between February 16, 2017, and March 12, 2018, 94 patients were randomly assigned (47 pembrolizumab/47 paclitaxel) after screening; enrollment was stopped on March 12, 2018, based on the results of the global KEYNOTE-061 study, and patients were followed until the last patient's last visit. Median OS was 8 months (95% confidence interval [CI], 4-10 months) with pembrolizumab versus 8 months (95% CI, 5-11 months) with paclitaxel (hazard ratio [HR], 0.99; 95% CI, 0.63-1.54). Median PFS was 2 months (95% CI, 1-3 months) with pembrolizumab versus 4 months (95% CI, 3-6 months) with paclitaxel (HR, 1.62; 95% CI, 1.04-2.52). ORR was 13% for pembrolizumab versus 19% for paclitaxel. Any-grade treatment-related adverse events occurred in 28 pembrolizumab-treated patients (60%) and 42 paclitaxel-treated patients (96%); grades 3 to 5 events occurred in 5 patients (11%) and 28 patients (64%), respectively. CONCLUSIONS: Definitive conclusions about the efficacy of second-line pembrolizumab in Asian patients with advanced PD-L1-positive gastric/GEJ cancer are limited because of insufficient power, but pembrolizumab was well tolerated in this patient population. Efficacy followed a trend similar to that observed in the phase 3 KEYNOTE-061 trial.

Intramolecular N-H⋠⋠⋠F Hydrogen Bonding Interaction in a Series of 4-Anilino-5-Fluoroquinazolines: Experimental and Theoretical Characterization of Electronic and Conformational Effects.

The 4-anilino-6,7-ethylenedioxy-5-fluoroquinazoline scaffold is presented as a novel model system for the characterization of the weak NH⋅⋅⋅F hydrogen bonding (HB) interaction. In this scaffold, the aniline NH proton is forced into close proximity with the nearby fluorine (dH,F ∼2.0 Å, ∠∼138°), and a through-space interaction is observed by NMR spectroscopy with couplings (1h JNH,F ) of 19±1 Hz. A combination of experimental (NMR spectroscopy and X-ray crystallography) and theoretical methods (DFT calculations) were used for the characterization of this weak interaction. In particular, the effects of conformational rigidity and steric compression on coupling were investigated. This scaffold was used for the direct comparison of fluoride with methoxy as HB acceptors, and the susceptibility of the NH⋅⋅⋅F interaction to changes in electron distribution and resonance was probed by preparing a series of molecules with different electron-donating or -withdrawing groups in the positions para to the NH and F. The results support the idea that fluorine can act as a weak HB acceptor, and the HB strength can be modulated through additive and linear electronic substituent effects.

Multi-omic biomarker identification and validation for diagnosing warzone-related post-traumatic stress disorder.

Post-traumatic stress disorder (PTSD) impacts many veterans and active duty soldiers, but diagnosis can be problematic due to biases in self-disclosure of symptoms, stigma within military populations, and limitations identifying those at risk. Prior studies suggest that PTSD may be a systemic illness, affecting not just the brain, but the entire body. Therefore, disease signals likely span multiple biological domains, including genes, proteins, cells, tissues, and organism-level physiological changes. Identification of these signals could aid in diagnostics, treatment decision-making, and risk evaluation. In the search for PTSD diagnostic biomarkers, we ascertained over one million molecular, cellular, physiological, and clinical features from three cohorts of male veterans. In a discovery cohort of 83 warzone-related PTSD cases and 82 warzone-exposed controls, we identified a set of 343 candidate biomarkers. These candidate biomarkers were selected from an integrated approach using (1) data-driven methods, including Support Vector Machine with Recursive Feature Elimination and other standard or published methodologies, and (2) hypothesis-driven approaches, using previous genetic studies for polygenic risk, or other PTSD-related literature. After reassessment of ~30% of these participants, we refined this set of markers from 343 to 28, based on their performance and ability to track changes in phenotype over time. The final diagnostic panel of 28 features was validated in an independent cohort (26 cases, 26 controls) with good performance (AUC = 0.80, 81% accuracy, 85% sensitivity, and 77% specificity). The identification and validation of this diverse diagnostic panel represents a powerful and novel approach to improve accuracy and reduce bias in diagnosing combat-related PTSD.

Comparison of the performance of risk scoring systems for tumor bleeding in patients with inoperable gastric cancer.

BACKGROUND: The Glasgow - Blatchford bleeding score (GBS) and admission and full Rockall scores are widely used risk scoring systems to stratify risk and determine the need for intervention in patients with upper gastrointestinal bleeding. We evaluated the performance of these risk scoring systems in patients with gastric cancer and tumor bleeding. METHODS: This retrospective study included patients with inoperable gastric cancer who presented with tumor bleeding at the National Cancer Center, Korea, between 2001 and 2015. The GBS, and admission and full Rockall scores were calculated. Primary outcome was the performance of the risk scoring systems in predicting the need for urgent interventions (endoscopic therapy, transarterial embolization, and surgery). The risk factors associated with urgent intervention were analyzed. RESULTS: Of 357 patients with tumor bleeding, 118 (33.1 %; 116 endoscopic therapy, 2 trans-arterial embolization) required urgent intervention. The full Rockall score was better at predicting the need for urgent intervention (area under the receiver operating characteristic curve = 0.78; P < 0.001) than the GBS (0.56) and admission Rockall score (0.56). Hemostatic intervention was not performed in patients with a full Rockall score ≤ 6 (103 patients, 28.9 %). On multivariate analysis, endoscopic stigmata of recent hemorrhage of Forrest classes Ia - IIb were significant factors associated with urgent intervention. CONCLUSIONS: The full Rockall score was superior to the GBS and admission Rockall score in predicting the need for urgent intervention for tumor bleeding. Thus, endoscopic evaluation is required to determine the need for urgent intervention in patients with gastric cancer and tumor bleeding.

Factors Influencing Motor Outcome of Hippotherapy in Children with Cerebral Palsy.

OBJECTIVE: This study was aimed to identify individual factors influencing the gross motor outcome of hippotherapy in children with cerebral palsy (CP). METHODS: One hundred and forty-six children with CP (mean age: 5.78 ± 1.72 years, male: 56.2%) presenting variable function (gross motor function classification system [GMFCS], levels I-IV) participated in this study. Participants received 30 minutes of hippotherapy twice a week for 8 weeks. Clinical information including GMFCS level, age, sex, CP distribution, CP type, gross motor function measure-88 (GMFM-88), GMFM-66, and pediatric balance scale (PBS) score were collected retrospectively. We regarded the children with GMFM-66 score increased by 2.0 points as good responders to hippotherapy. Further we analyzed factors affecting good responders. RESULTS: GMFCS level I and II compared with IV (odds ratio [OR] = 6.83) and III compared with IV (OR = 4.45) were significantly associated with a good response to hippotherapy. Higher baseline GMFM E (OR = 1.05) and lower baseline GMFM B (OR = 0.93) were also significantly associated with a good response to hippotherapy. Sex, age, CP type, and distribution were not factors influencing gross motor outcome of hippotherapy. CONCLUSIONS: The children with CP, GMFCS level I-III, with relatively poor postural control in sitting might have a greater chance to improve their GMFM-66 scores through hippotherapy. This supports the hypothesis that hippotherapy is a context-focused therapy to improve postural control in sitting.

Review of the effect of 3D medical printing and virtual reality on urology training with 'MedTRain3DModsim' Erasmus + European Union Project

Background/aim: It is necessary to incorporate novel training modalities in medical education, especially in surgical fields, because of the limitations of cadaveric training. Traditional medical education has many drawbacks, such as residency working hour restrictions, patient safety conflicts with the learning needs, and the lack of hands-on workshops. The MedTRain3DModsim Project aimed to produce 3-dimensional (3D) medical printed models, simulations, and innovative applications for every level of medical training using novel worldwide technologies. It was aimed herein to improve the interdisciplinary and transnational approaches, and accumulate existing experience for medical education, postgraduate studies, and specialty training. Materials and methods: This project focused on models of solid organs and the urinary system, including the kidney, prostate, ureter, and liver. With 3D medical printing, it is possible to produce a body part from inert materials in just a few hours with the standardization of medical 3D modeling. Results: The target groups of this project included medical students and residents, graduate students from engineering departments who needed medical education and surgical training, and medical researchers interested in health technology or clinical and surgical an atomy. Conclusion: It was also intended to develop a novel imaging platform for education and training by reevaluating the existing data using new software and 3D modalities. Therefore, it was believed that our methodology could be implemented in all related medical fields.

Radiosynthesis, biological evaluation and preliminary microPET study of 18F-labeled 5-resorcinolic triazolone derivative based on ganetespib targeting HSP90.

Heat-shock protein 90 (HSP90) is a molecular chaperone that activates oncogenic transformation in several solid tumors, including lung and breast cancers. Ganetespib, a most promising candidate among several HSP90 inhibitors under clinical trials, has entered Phase III clinical trials for cancer therapy. Despite numerous evidences validating HSP90 as a target of anticancer, there are few studies on PET agents targeting oncogenic HSP90. In this study, we synthesized and biologically evaluated a novel 18F-labeled 5-resorcinolic triazolone derivative (1, [18F]PTP-Ganetespib) based on ganetespib. [18F]PTP-Ganetespib was labeled by click chemistry of Ganetespib-PEG-Alkyne (10) and [18F]PEG-N3 (11) with 37.3 ±â€¯5.11% of radiochemical yield and 99.7 ±â€¯0.09% of radiochemical purity. [18F]PTP-Ganetespib showed proper LogP (0.96 ±â€¯0.06) and good stability in human serum over 97% for 2 h. [18F]PTP-Ganetespib showed high uptakes in breast cancer cells containing triple negative breast cancer (TNBC) MDA-MB-231 and Her2-negative MCF-7 cells, which are target breast cancer cell lines of HSP90 inhibitor, ganetespib, as an anticancer. Blocking of HSP90 by the pretreatment of ganetespib exhibited significantly decreased accumulation of [18F]PTP-Ganetespib in MDA-MB-231 and MCF-7 cells, indicating the specific binding of [18F]PTP-Ganetespib to MDA-MB-231 and MCF-7 cells with high HSP90 expression. In the biodistribution and microPET imaging studies, the initial uptake into tumor was weaker than in other thoracic and abdominal organs, but [18F]PTP-Ganetespib was retained relatively longer in the tumor than other organs. The uptake of [18F]PTP-Ganetespib in tumors was not sufficient for further development as a tumor-specific PET imaging agent by itself, but this preliminary PET imaging study of [18F]PTP-Ganetespib can be basis for developing new PET imaging agents based on HSP90 inhibitor, ganetespib.

Efficacy and tolerability of ramucirumab monotherapy or in combination with paclitaxel in gastric cancer patients from the Expanded Access Program Cohort by the Korean Cancer Study Group (KCSG).

BACKGROUND: Ramucirumab improves survival in gastric cancer patients. The efficacy and safety of ramucirumab outside of a clinical trial were evaluated using an expanded access program (EAP). METHODS: Advanced gastric cancer patients treated with ramucirumab in combination with paclitaxel or with ramucirumab monotherapy in a Korean EAP were evaluated. Baseline characteristics were assessed for progression-free survival (PFS) and overall survival (OS), and adverse events were evaluated according to the treatment regimen. RESULTS: Of 265 patients, 228 received ramucirumab plus paclitaxel, and 37 received ramucirumab monotherapy. Grade 3 or 4 neutropenia was more common with ramucirumab plus paclitaxel than with ramucirumab monotherapy (46.7 vs. 8.1%). Gastrointestinal (GI) perforation developed in seven patients (3.1%) in the ramucirumab plus paclitaxel group. The overall response and disease control rates were 16.6 and 66.3% in the ramucirumab plus paclitaxel group, and 5.4 and 37.8% in the ramucirumab monotherapy group, respectively. PFS and OS were 3.8 and 8.6 months in the ramucirumab plus paclitaxel group, and 1.8 and 6.4 months in the ramucirumab monotherapy group, respectively. In multivariate analysis, alkaline phosphatase, albumin, and neutrophil-to-lymphocyte ratio (NLR) were the independent prognostic factors for PFS, while albumin, NLR, number of metastatic sites, and large amount of ascites were independent prognostic factors for OS. CONCLUSION: In the Korean EAP cohort, ramucirumab showed similar efficacy to the results of the previous trials for gastric cancer. However, the level of GI perforation was slightly increased in the ramucirumab plus paclitaxel group.

Factorial Validity and Measurement Invariance of the Test of Preschool Early Literacy-Phonological Awareness Test Among Deaf and Hard-of-Hearing Children and Hearing Children.

OBJECTIVES: Emerging evidence suggests that early phonological awareness in deaf and hard-of-hearing (DHH) children with functional hearing is significantly related to their reading acquisition, and the assessment of phonological awareness can play a critical role in preventing reading difficulties. Validation of the scores obtained from standardized assessments when used with DHH students is crucial to support the assessments' intended interpretations and implications of test scores. Using archival data sets, the aim of this study was twofold: (a) to establish the factorial validity of the item scores on the Test of Preschool Early Literacy-Phonological Awareness (TOPEL-PA) for DHH children with functional hearing and hearing children and (b) to test measurement invariance across these groups. Our archival data sets included assessments of DHH children, hearing children from low socioeconomic status (SES) backgrounds, and hearing children from a range of SES backgrounds. We hypothesized that a second-order unifying ability, Phonological Awareness, along with four first-order subtest factors would explain inter-item associations among the 27 items on the TOPEL-PA. We further hypothesized that patterns of associations among the item scores would be similar across groups and that the individual items would function similarly across groups. DESIGN: Seven hundred and thirty-three children from three samples participated in the study; 171 were DHH children (Mage = 58.7 months old, SDage = 12.5 months old), 195 were low-SES hearing children (Mage = 55.5 months old, SDage = 3.5 months old), and 367 were diverse-SES hearing children (Mage = 53.4 months old, SDage = 8.9 months old). All DHH children were able to identify the referent of monosyllabic spoken words on the Early Speech Perception Test. RESULTS: Test of confirmatory item factor analyses of the hypothesized second-order factor structure revealed that a second-order unifying ability along with four first-order subtest factors well explained associations among the item scores for all groups. This aligned with the scoring structure of the TOPEL-PA, providing strong evidence for factorial validity of the item scores for DHH children as well as for hearing children groups. The measurement invariance test results provided evidence that the vast majority of TOPEL-PA items functioned similarly for hearing children and DHH children with speech perception abilities, suggesting that the utility of the assessment scores obtained from DHH children is consistent with the scores obtained from hearing children. CONCLUSION: Results of our study suggest that researchers and practitioners can use the TOPEL-PA to assess DHH children with functional hearing. It also suggests that the two skills measured on the TOPEL-PA (blending and elision) are qualitatively similar for DHH and hearing children, but the latent mean score obtained from the DHH children significantly differed from those of the hearing groups, suggesting a quantitative difference.

Chemical characteristics and enhanced hepatoprotective activities of Maillard reaction products derived from milk protein-sugar system.

The objective of this study was to investigate the characteristics, antioxidative properties, and hepatoprotective effects of Maillard reaction products (MRP) from milk protein reacted with sugars. The MRP were obtained from milk protein, whey protein concentrates and sodium caseinate, using 2 types of sugars, lactose and glucose, by heating the mixture at 55°C for 7d in a sodium phosphate buffer (pH 7.4). Changes in the chemical modification of the milk protein were monitored by measuring the protein-bound carbonyls and PAGE protein profiles. The results showed that the amount of protein-bound carbonyls increased after Maillard reaction (MR). In addition, sodium dodecyl sulfate-PAGE analysis indicated a formation of high-molecular weight complexes through MR. The modification sites induced by MR of milk protein were monitored by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of tryptic-digested gel spots of MRP. As a result, modification and their localization in AA sequence of MRP was identified. Also, the MRP showed higher antioxidant activities than the intact milk protein, and they reduced intracellular reactive oxygen species production and inhibited the depletion of the reduced glutathione concentrations in the HepG2 cells. In particular, glucose-sodium caseinate MRP showed the highest biological activities among all MRP. Therefore, these results suggest that the MRP from milk protein reacting with sugars possess effective antioxidant activity and have a protective ability against oxidative damage.

Biophysical mechanisms of traumatic brain injuries.

Despite years of effort to prevent traumatic brain injuries (TBIs), the occurrence of TBI in the United States alone has reached epidemic proportions. When an external force is applied to the head, it is converted into stresses that must be absorbed into the brain or redirected by a helmet or other protective equipment. Complex interactions of the head, neck, and jaw kinematics result in strains in the brain. Even relatively mild mechanical trauma to these tissues can initiate a neurochemical cascade that leads to TBI. Civilians and warfighters can experience head injuries in both combat and noncombat situations from a variety of threats, including ballistic and blunt impact, acceleration, and blast. It is critical to understand the physics created by these threats to develop meaningful improvements to clinical care, injury prevention, and mitigation. Here the authors review the current state of understanding of the complex loading conditions that lead to TBI and characterize how these loads are transmitted through soft tissue, the skull and into the brain, resulting in TBI. In addition, gaps in knowledge and injury thresholds are reviewed, as these must be addressed to better design strategies that reduce TBI incidence and severity.

Relation of repeated low-level blast exposure with symptomology similar to concussion.

OBJECTIVE: To investigate anecdotal reports suggesting that repeated exposure to low-level explosive blast has myriad health impacts, including an array of neurological effects. PARTICIPANTS: A total of 184 anonymous survey respondents from military and nonmilitary law enforcement populations (135 exposed to occupational blast and 49 controls). DESIGN: Survey of self-reported history of occupational exposure to repeated low-level blast (breaching blast) and symptomology similar to concussion. RESULTS: Findings suggest that number and severity of symptoms increase with history of chronic blast exposure (F = 18.26, P < .001) and that symptoms can interfere with daily activity (t = 2.60, P = .010). CONCLUSION: Given the prevalence of repeated exposure to blast among some military and civilian law enforcement occupations, the results of this survey study support a role for blast surveillance programs as well as continued research on health impacts of low-level repeated blast exposure.

Suppression of splenic lymphocyte proliferation by Eucommia ulmoides and genipin.

We investigated the modulation of innate and adaptive immune cell activation by Eucommia ulmoides Oliver extract (EUE) and its ingredient genipin. As an innate immunity indicator, the phagocytic activity of macrophages was determined by measuring engulfed, fluorescently labeled Escherichia coli. As a surrogate marker for the respective activation of cellular and humoral adaptive immunity, concanavalin A (Con A) and lipopolysaccharide (LPS) induction of primary splenocyte proliferation was assayed in in vitro and ex vivo systems. EUE and genipin suppressed the proliferation of primary splenic lymphocytes induced by Con A or LPS, but not macrophage phagocytosis. Oral administration of EUE and genipin to mice decreased splenic lymphocyte proliferation induced by Con A or LPS. These results revealed that E. ulmoides and genipin suppressed cellular and humoral adaptive immunity, and they suggest that E. ulmoides and genipin are promising candidates for immunosuppressive drugs that target diseases that involve excessive activation of adaptive immunity.

In-vitro study on ureteral smooth muscle contractility with tamsulosin, nifedipine, and terpene mixture (Rowatinex®).

AIM: The aim of this study was to evaluate whether tamsulosin, an alpha-blocker, has an effect on decreasing spontaneous ureteral contractility with or without phenylephrine, an alpha-agonist. Additionally, nifedipine and a terpene mixture (Rowatinex®) were tested and compared with each other. METHODS: We obtained ureteral segments from freshly killed eight-week-old rabbits. Preparation was performed in an aerated Krebs buffer (95% oxygen and 5% carbon dioxide) at a constant temperature of 37 °C. All segments were suspended into organ tissue baths containing aerated Krebs buffer using stainless steel hangers and clips. The ureter was divided into four segments: upper, middle, low and uretero-vesical junction. Each ureteral segment was suspended longitudinally and circularly by opposite corners, respectively. Tamsulosin, nifedipine, and the terpene mixture were separately applied into the segments. Contractile activities of each drug were recorded and analyzed by the PowerLab data acquisition system (AD instruments CO., USA). The area under the curve was compared between before and after each drug application for each 5 minutes with or without pheylephrine. Statistical analysis was performed using the unpaired Student's t test. RESULTS: Under Krebs solution, ureteral smooth muscle contractility was significantly decreased in all segments over 10(-6) M in tamsulosin, 10(-7) M in nifedipine and 0.001x1 concentrations in the terpene mixture (P=0.038). However, under Krebs solution with 10(-5) M phenylephrine, there was no significant difference at all concentrations in tamsoluin and nifedipine. In contrast to tamsolusin and nifedipine, there was a significant decrease in ureteral smooth muscle contractility in most of segments at 0.01x1 concentrations (P=0.042) in the terpene mixture. CONCLUSION: Tamsulosin, nifedipine, and the terpene mixture showed the effect on spontaneous ureteral contractility. In particular, the terpene mixture might have the better effect on decreasing ureteral smooth muscle contractility.

The heat-shock protein-70-induced renoprotective effect is partially mediated by CD4+ CD25+ Foxp3 + regulatory T cells in ischemia/reperfusion-induced acute kidney injury.

Recent reports suggest the presence of heat-shock protein (HSP)-reactive T cells with a regulatory phenotype in various inflammatory diseases. To test whether HSP exerts renoprotective effects through regulatory T cells (Tregs), ischemia/reperfusion injury was done with or without heat preconditioning in mice. Splenocytes from heat-preconditioned mice had Treg expansion and a reduced proliferative response upon mitogenic stimulus. T cells from heat-preconditioned mice failed to reconstitute postischemic injury when adoptively transferred to T cell-deficient nu/nu mice in contrast to those from control mice. Tregs were also increased in heat-preconditioned ischemic kidneys. Depleting Tregs before heat preconditioning abolished the renoprotective effect, while adoptive transfer of these cells back into Treg-depleted mice partially restored the beneficial effect of heat preconditioning. Inhibition of HSP70 by quercetin suppressed Treg expansion, as well as renoprotective effects. Transferring Tregs in quercetin-treated heat-preconditioned mice partially restored the beneficial effect of heat preconditioning. The specificity of immune cell HSP70 in renoprotection was confirmed by partial restoration of kidney injury when T cells from HSP70-deficient heat preconditioned mice were adoptively transferred to nu/nu mice. Thus, the renoprotective effect of HSP70 may be partially mediated by a direct immunomodulatory effect through Tregs. Better understanding of immunomodulatory mechanisms of various stress proteins might facilitate discovery of new preventive strategies in acute kidney injury.

Outcomes of second self-expandable metallic stent insertion for malignant gastric outlet obstruction.

BACKGROUND: Self-expandable metallic stents are used widely to relieve malignant gastric outlet obstruction (GOO). However, restenosis or migration of first stents is a frequent complication. The purpose of this retrospective cohort study was to evaluate the effectiveness of second stents as an approach to manage failure of first stents in patients with malignant GOO. METHODS: A total of 222 patients with gastric cancer received first stents due to inoperable GOO at National Cancer Center in Korea between January 2008 and June 2011. Monthly follow-up interviews were performed, and second stents (stent-in-stent or stent-after-migration) were inserted in 59 patients by June 2012. Technical and clinical successes and long-term complications were evaluated. RESULTS: The technical and immediate clinical success rates were 98.3 % (58/59) and 91.5 % (54/59), respectively. Patients who received a second stent due to late complications involving the first stent (migration, restenosis, and fracture) showed a higher clinical success rate (95.8 % [46/48]) than patients who received a second stent due to immediate clinical failure of the first stent (72.7 % [8/11], p = 0.04). The immediate clinical success rate of stent-after-migration (100 % [11/11]) was not different from that of stent-in-stent (89.6 % [43/48], p = 1.0). The stent dysfunction rate of stent-after-migration (27.3 % [3/11]) also was similar to that of stent-in-stent (29.2 % [14/48], p = 1.0). The median patencies of stent-in-stent and stent-after-migration were 27.4 and 58.4 weeks, respectively (p = 0.177). There were no significant prognostic factors for patency of second stents. CONCLUSIONS: Insertion of a second stent is effective for treating the first-stent failure in gastric cancer patients with GOO, especially if the immediate outcome of the first stent was successful.

Enhanced toxicity-free astaxanthin extraction from Haematococcus pluvialis via concurrent cell disruption and demulsification.

The extraction of astaxanthin from Haematococcus pluvialis involves the utilization of petroleum-derived organic solvents or supercritical CO2, beset by safety concerns, high costs, and environmental sustainability limitations. This study, in contrast, employed a method involving the adjustment of salt concentration, propylene glycol, and vegetable oil fraction to disrupt emulsion in aqueous cell lysates for facilitating the separation of astaxanthin. Under optimized conditions, an astaxanthin-containing oil with a content of 1.88% was obtained even with the use of wet biomass, and four rounds of consecutive extraction resulted in a cumulative recovery yield of 66.41%. This process produced astaxanthin-enriched soybean oil with 9.49 times improved antioxidant capacity that satisfies a requirement for health functional application. Omitting the solvent removal and drying processes, which consume tremendous energy, can reduce the production cost by 2.98 times compared to conventional methods. Consequently, this study suggests an effective technique for producing edible oil containing H. pluvialis-derived astaxanthin.

Relationship between rheological properties, sensory perception, and overall acceptability of thickened liquids for dysphagia in young and old healthy individuals.

This study investigated the relationship between rheological properties, sensory perception, and overall acceptability in healthy young and old groups for dysphagia thickened liquids. Unflavored (UTL) and flavored (FTLP) thickened liquids were prepared using tap water or pomegranate juice at 10 different viscosity levels. The rheological properties were then evaluated via syringe flow test and line spread test (LST). When the apparent viscosity levels of UTL and FTLP were similar, the syringe test and LST results were also similar, indicating consistent flow behavior. Sensory perception evaluations showed that the young group better distinguished viscosity differences between stages compared to the old group. Regarding overall acceptability, the old group preferred samples with higher apparent viscosity than the young group. Principal component analysis and k-means cluster analysis were used to explore correlations between variables and classify thickened liquids into four groups. This can serve the foundation for standardized texture grades of dysphagia thickened liquids, considering rheological characteristics and sensory profiles.

Generation of a human SLC12A3 knock-in human induced pluripotent stem cell line (CMCi014-A-82) using CRISPR-Cas9 system.

Gitelman's disease is caused by a genetic mutation in the solute carrier family 12 member 3 (SLC12A3) gene, which encodes the sodium chloride cotransporter. In this study, we generated a stable human induced pluripotent stem cell (hiPSC) line, WTC11-SLC12A3 (CMCi014-A-82), by knocking in the entire SLC12A3 gene at the SHS231 locus in healthy wild-type control hiPSCs (WTC11). We verified that WTC11-SLC12A3 expressed pluripotency markers and exhibited normal stem cell morphology. Furthermore, this cell line maintains a normal karyotype and can differentiate into the three germ layers. Therefore, this cell line may provide a basis for gene therapy for Gitelman's disease.