TGFß2-Driven Ferritin Degradation and Subsequent Ferroptosis Underlie Salivary Gland Dysfunction in Postmenopausal Conditions.

Despite the high incidence of dry mouth in postmenopausal women, its underlying mechanisms and therapeutic interventions remain underexplored. Using ovariectomized (OVX) mouse models, here this study identifies ferroptosis, an iron-dependent regulated cell death, as a central mechanism driving postmenopausal salivary gland (SG) dysfunction. In the OVX-SGs, TGFß signaling pathway is enhanced with the aberrant TGFß2 expression in SG mesenchymal cells. Intriguingly, TGFß2 treatment reduces iron-storing ferritin levels, leading to lipid peroxidation and ferroptotic death in SG epithelial organoids (SGOs). Mechanistically, TGFß2 promotes the autophagy-mediated ferritin degradation, so-called ferritinophagy. A notable overexpression of the type III TGFß receptor (TßRIII) is found in the OVX-SGs and TGFß2-treated SGOs, while the silencing of TßRIII mitigates the ferroptosis-mediated deleterious effects of TGFß2 on SGOs. Finally, administration of ferroptosis inhibitor, Liproxstatin-1 (Lip-1), improves saliva secretion in OVX mice. Present findings collectively suggest a link between TGFß signaling, ferroptosis, and SG injury, offering new therapeutic avenues for postmenopausal xerostomia.

Sustained release of stem cell secretome from nano-villi chitosan microspheres for effective treatment of atopic dermatitis.

Canine atopic dermatitis (AD) arises from hypersensitive immune reactions. AD symptoms entail severe pruritus and skin inflammation, with frequent relapses. Consequently, AD patients require continuous management, imposing financial burdens and mental fatigue on pet owners. In this study, we aimed to investigate the therapeutic relevance of secretome from canine adipose tissue-derived mesenchymal stem cells (MSCs), especially after encapsulation in nano-villi chitosan microspheres (CS-MS) to expect improved efficacy. Conditioned media (CM) from MSCs significantly inhibited the proliferation of splenocytes, induced the generation of regulatory T cells, and decreased mast cell degranulation. We found that beneficial soluble factors known to reduce AD symptoms, including transforming growth factor-beta 1, were detectable after sequential concentration and lyophilization of CM. The CS-MS, developed by a phase inversion regeneration method, showed high loading and sustained release of the secretome. Local injection of secretome-loaded CS-MS (ST/SC-MS) effectively reduced clinical severity compared to groups treated with secretome. Histological analysis revealed that ST/SC-MS potently suppressed epidermal hyperplasia, immunocyte infiltration and mast cell activation in the lesion. Taken together, this study presents a novel therapeutic approach exhibiting more potent and prolonged immunoregulatory efficacy of MSC secretome for canine AD treatment.

Hexane extract of green tea (Camellia sinensis) leaves is an exceptionally rich source of squalene.

Squalene is a cancer chemo-preventive and skin protective agent with high commercial demand. Here, we report for the first time that the green tea (Camellia sinensis) leaves is a surprisingly rich plant-based source of squalene. Young and tender leaves and old and turf leaves were collected at four different collecting seasons (April-August). Lipophilic compounds in the leaves and commercial green teas were extracted with hexane. The squalene contents in the hexane extracts varied greatly with the types of the leaves and collecting seasons. The hexane extract of turf leaves contained significantly higher contents of squalene than the extract of tender leaves. The hexane extract of the turf leaves collected in August contained the highest content of squalene (29.2 g/kg extract). This represents the first report on the qualitative and quantitative information on squalene in green tea leaves.

Comparison of One- and Two-Region of Interest Strain Elastography Measurements in the Differential Diagnosis of Breast Masses.

OBJECTIVE: To compare the diagnostic performance and interobserver variability of strain ratio obtained from one or two regions of interest (ROI) on breast elastography. MATERIALS AND METHODS: From April to May 2016, 140 breast masses in 140 patients who underwent conventional ultrasonography (US) with strain elastography followed by US-guided biopsy were evaluated. Three experienced breast radiologists reviewed recorded US and elastography images, measured strain ratios, and categorized them according to the American College of Radiology breast imaging reporting and data system lexicon. Strain ratio was obtained using the 1-ROI method (one ROI drawn on the target mass), and the 2-ROI method (one ROI in the target mass and another in reference fat tissue). The diagnostic performance of the three radiologists among datasets and optimal cut-off values for strain ratios were evaluated. Interobserver variability of strain ratio for each ROI method was assessed using intraclass correlation coefficient values, Bland-Altman plots, and coefficients of variation. RESULTS: Compared to US alone, US combined with the strain ratio measured using either ROI method significantly improved specificity, positive predictive value, accuracy, and area under the receiver operating characteristic curve (AUC) (all p values < 0.05). Strain ratio obtained using the 1-ROI method showed higher interobserver agreement between the three radiologists without a significant difference in AUC for differentiating breast cancer when the optimal strain ratio cut-off value was used, compared with the 2-ROI method (AUC: 0.788 vs. 0.783, 0.693 vs. 0.715, and 0.691 vs. 0.686, respectively, all p values > 0.05). CONCLUSION: Strain ratios obtained using the 1-ROI method showed higher interobserver agreement without a significant difference in AUC, compared to those obtained using the 2-ROI method. Considering that the 1-ROI method can reduce performers' efforts, it could have an important role in improving the diagnostic performance of breast US by enabling consistent management of breast lesions.

Synthesis and formation mechanism of bone mineral, whitlockite nanocrystals in tri-solvent system.

Whitlockite (WH, Ca18Mg2(HPO4)2(PO4)12) is the second most abundant bone mineral and has attracted attention as one of the novel bone regenerative materials. It has proven to enhance growth and promote osteogenesis of stem cells. However, investigating the mechanism of formation of pure phase WH nanocrystals remains a challenge. In this study, we introduced an interesting synthesis approach of WH nanocrystals using a tri-solvent system for the solid-liquid-solution (SLS) process. The ratio of precursor and reaction solvent composition was optimized to generate WH nanocrystals with tunable size, morphology (nanoplates, nanospheres), and surface properties (hydrophobic, hydrophilic), which is impossible to achieve using the traditional precipitation method. Molecular dynamics (MD) simulations revealed that the growth direction of nanoplates is highly related to the surfactant and its binding affinity. Finite element method (FEM) simulations elucidated that the ratio of ethanol/water plays an important role in defining the crystallinity and morphology of WH. In this study, we demonstrated that the cell proliferation of human bone marrow-derived mesenchymal stem cells (hBMSCs) is enhanced upon treatment with WH. The results of quantitative real-time polymerase chain reaction (qPCR) revealed that WH can positively accelerate the osteogenic differentiation in hBMSCs. The as-synthesized WH has a great potential in the future to be used in osteogenic tissue engineering. This study opens a new horizon for the synthesis and application of WH.

Clinical and Computed Tomography Angiographic Predictors of Coronary Lesions That Later Progressed to Chronic Total Occlusion.

OBJECTIVES: This study aimed to investigate clinical and coronary computed tomographic angiography (CTA) characteristics of lesions that progressed to chronic total occlusion (CTO). BACKGROUND: CTO is one of the most common reasons for referral to coronary artery bypass surgery. Prediction and adequate early management for future CTO lesions may be beneficial. METHODS: The study evaluated patients with at least 1 vessel with a diameter stenosis of ≥70% on invasive coronary angiography (ICA) who underwent previous coronary CTA >12 months before ICA, from 2006 to 2015. The study compared the baseline clinical and coronary CTA characteristics of the patients with future CTO lesions with those of the patients with future non-CTO lesions (patient-level analysis) and compared coronary CTA findings between the future CTO lesion with the most stenotic non-CTO lesion in each CTO patient (lesion-level analysis). RESULTS: Among the 216 patients, 32 (14.8%) had a CTO lesion on ICA. In patient-level analysis, no significant differences in clinical characteristics were found, whereas the coronary CTA culprit lesions of the CTO group had a smaller minimal lumen diameter (MLD) with more adverse plaque characteristics. In lesion-level analysis, future CTO lesions had a smaller MLD, a smaller reference segment diameter (RD), and longer lesion length. These lesions were more likely to be noncalcified plaques with a noneccentric cross-sectional distribution, and had a higher remodeling index, lower mean plaque attenuation (MPA), and more napkin-ring signs. In multivariate analysis and receiver-operating characteristic curve analysis, MLD of <2.0 mm, RD of <3.2 mm, and MPA of <50 Hounsfield units were independent predictors of future CTO lesions. The risk of CTO development in lesions with triple risk factors was 14-fold higher than that of the lesions with no risk factors. CONCLUSIONS: Lesions that progressed to CTO had more severe baseline coronary CTA features than non-CTO lesions. A small MLD, small RD, and low MPA were independent predictors of progression to CTO.

A computer-aided diagnosis system using artificial intelligence for the diagnosis and characterization of breast masses on ultrasound: Added value for the inexperienced breast radiologist.

To evaluate the value of the computer-aided diagnosis (CAD) program applied to diagnostic breast ultrasonography (US) based on operator experience.US images of 100 breast masses from 91 women over 2 months (from May to June 2015) were collected and retrospectively analyzed. Three less experienced and 2 experienced breast imaging radiologists analyzed the US features of the breast masses without and with CAD according to the Breast Imaging Reporting and Data System (BI-RADS) lexicon and categories. We then compared the diagnostic performance between the experienced and less experienced radiologists and analyzed the interobserver agreement among the radiologists.Of the 100 breast masses, 41 (41%) were malignant and 59 (59%) were benign. Compared with the experienced radiologists, the less experienced radiologists had significantly improved negative predictive value (86.7%-94.7% vs 53.3%-76.2%, respectively) and area under receiver operating characteristics curve (0.823-0.839 vs 0.623-0.759, respectively) with CAD assistance (all P < .05). In contrast, experienced radiologists had significantly improved specificity (52.5% and 54.2% vs 66.1% and 66.1%) and positive predictive value (55.6% and 58.5% vs 64.9% and 64.9%, respectively) with CAD assistance (all P < .05). Interobserver variability of US features and final assessment by categories were significantly improved and moderate agreement was seen in the final assessment after CAD combination regardless of the radiologist's experience.CAD is a useful additional diagnostic tool for breast US in all radiologists, with benefits differing depending on the radiologist's level of experience. In this study, CAD improved the interobserver agreement and showed acceptable agreement in the characterization of breast masses.

Assessment of a new genomic classification system in acute myeloid leukemia with a normal karyotype.

This study was performed to assess if a recently recommended genomic classification is predictive in patients with normal-karyotype (NK) acute myeloid leukemia (AML). A total of 393 patients were included. Analysis of genetic mutations was performed using targeted resequencing with an Illumina Hiseq 2000. We identified driver mutations across 40 genes, with one or more driver mutations identified in 95.7% of patients. The molecular subclassification was as follows: 34.6% patients (n = 136) with AML with the NPM1 mutation, 10.7% (n = 42) with AML with mutated chromatin or RNA-splicing genes or both, 1.5% (n = 6) with AML with TP53 mutations, 13.5% (n = 53) with AML with biallelic CEBPA mutations, 2.0% (n = 8) with AML with IDH2-R172 mutations and no other class-defining lesion, 29.5% (n = 116) with AML with driver mutations but no detected class-defining lesion, 4.3% (n = 17) with AML with no detected driver mutation, and 3.8% (n = 15) patients with AML who met the criteria for ≥2 genomic subgroups. The 5-year overall survival and relapse rate of subgroup in AML with mutated chromatin, RNA-splicing genes, or both was 11.6% (95% CI = 1.4-21.8%) and 71.4% (95% CI = 45.7-86.5%), respectively. This study suggests that the recently recommended genomic classification is an appropriate and replicable categorization system in the NK AML population. The subgroup of AML with mutated chromatin, RNA-splicing genes, or both showed extremely poor survival in NK-AML; thus, a novel approach is needed to improve their prognosis.

Early stimulated thyroglobulin for response prediction after recombinant human thyrotropin-aided radioiodine therapy.

OBJECTIVE: Measurement of recombinant human thyrotropin (rhTSH)-stimulated thyroglobulin (Tg) is generally recommended 72 h after the second rhTSH injection. However, due to the acute effect of I-131 on thyrocytes, Tg measured after radioiodine therapy (RIT) would not accurately reflect the thyroid tissue burden. We aimed to determine predictive values of serum Tg level measured just before rhTSH-aided RIT and to compare the results obtained just after RIT in patients with differentiated thyroid carcinoma (DTC). METHODS: We evaluated 150 patients with DTC who underwent rhTSH-aided RIT (2.96-6.66 GBq) after total thyroidectomy between 2009 and 2014. Serum Tg level was measured 24 h (early Tg) and 72 (or 96) h (delayed Tg) after the second rhTSH injection. An excellent response was defined based on the latest American Thyroid Association Guidelines. Univariate and multivariate analyses were performed for early Tg, delayed Tg, and other clinical variables. RESULTS: In the multivariate analysis, tumor size [odds ratio (OR) 1.716; 95% confidence interval (CI) 1.019-2.882; p = 0.042] and early Tg level (OR 2.012; 95% CI 1.384-2.925, p < 0.001) independently predicted excellent responses. The cutoff for the best early Tg level to predict a non-excellent response was 2.0 ng/mL. Delayed Tg was not a significant predictor (OR 0.992; 95% CI 0.969-1.015; p = 0.492). CONCLUSIONS: Early stimulated Tg significantly predicted therapeutic response after rhTSH-aided RIT in patients with DTC. Therefore, serum Tg should be measured before RIT to predict therapeutic responses.

5-Hydroxymethylcytosine correlates with epigenetic regulatory mutations, but may not have prognostic value in predicting survival in normal karyotype acute myeloid leukemia.

Stem cells display remarkably high levels of 5-hydroxymethylcytosine (5hmC). Both TET2 and IDH1/2 mutations can impair the production of 5hmC, thus decreasing 5hmC levels. TET2 or IDH1/2 mutations are commonly observed in acute myeloid leukemia (AML). However, the implications of 5hmC on survival in normal karyotype AML patients have not been fully evaluated. The 5hmC levels were analyzed in 375 patients using ELISA. The levels of 5hmC in DNA samples were converted to a log scale for the analysis and correlations with TET2 and/or IDH1/2 mutations were evaluated. The median 5hmC level was 0.065% (range 0.001-0.999). Mutation rates were 13.1% for TET2mut, 6.7% for IDH1mut, and 13.9% for IDH2mut. The prevalence of TET2 and/or IDH1/2 was 33.1% (124/375). TET2 and IDH1/2 mutated patients had significantly lower levels of log(5hmC) compared with patients without TET2 or IDH1/2 mutations (p<0.001). With a median follow-up of 55.5 months (range, 0.7-179.8), there was no significant difference in overall survival, event-free survival, and relapse risk according to TET2mut or IDH1/2mut (all, p>0.05). To identify its prognostic value, we sub-classified the levels of 5hmC into tertiles for 5hmC values. However, there was no significant association between the categories of 5hmC levels and survival or relapse risk (all p>0.05). Patients with TET2 or IDH1/2 mutations had lower levels of 5hmC. The 5hmC levels may not be predictive of survival in patients with normal karyotype AML.

Spectrum of somatic mutation dynamics in chronic myeloid leukemia following tyrosine kinase inhibitor therapy.

Somatic mutations commonly detected in a variety of myeloid neoplasms have not been systematically investigated in chronic myeloid leukemia (CML). We performed targeted deep sequencing on a total of 300 serial samples from 100 CML patients; 37 patients carried mutations. Sixteen of these had evidence of mutations originating from preleukemic clones. Using unsupervised hierarchical clustering, we identified 5 distinct patterns of mutation dynamics arising following tyrosine kinase inhibitor (TKI) therapy. This study demonstrates that patterns of mutation acquisition, persistence, and clearance vary but have a number of interesting correlations with clinical outcomes. Mutation burden often persisted despite successful TKI response (pattern 1), providing indirect evidence that these mutations also originated from preleukemic mutations, whereas patients exhibiting mutation clearance (pattern 3) showed mixed clinical outcomes. Unsurprisingly, patients acquiring new mutations during treatment failed TKI therapy (pattern 2). These patterns show that CML mutation dynamics following TKI therapy are markedly distinct from other myeloid neoplasms. In summary, clinical implications of mutation profiles and dynamics in CML should be interpreted with caution.

Iodine Uptake Patterns on Post-ablation Whole Body Scans are Related to Elevated Serum Thyroglobulin Levels After Radioactive Iodine Therapy in Patients with Papillary Thyroid Carcinoma.

PURPOSE: Serum thyroglobulin (Tg) level is frequently elevated shortly after radioactive iodine (RAI) ablation therapy. The authors studied the relationship between the elevation of serum Tg after RAI therapy and iodine uptake pattern on post-ablation whole body scans (RxWBSs) in patients with papillary thyroid carcinoma (PTC). MATERIALS AND METHODS: The study subjects were patients with PTC that had undergone first RAI therapy with thyroid hormone withdrawal after total thyroidectomy. Patients with a high level of serum anti-Tg antibody (TgAb, ≥ 60 U/mL), possible regional or distant metastasis as determined by pre-ablation or post-ablation studies, and negative iodine uptake of the anterior neck on RxWBS were excluded. Serum Tg was checked twice, that is, 7 days after (post-ablation Tg) and on the day of RAI therapy (pre-ablation Tg). Ratio of pre-ablation Tg to post-ablation Tg (Tg ratio) was used to assess changes in serum Tg levels after RAI therapy. Patients were classified into two groups according to the presence of midline uptake above the thyroidectomy bed on RxWBS (negative (group 1) or positive (group 2) midline uptake). Variables were subjected to analysis to identify differences between the two groups. RESULTS: Two hundred and fifty patients were enrolled in this study; 101 in group 1 and 149 in group 2. Based on univariate analysis, post-ablation Tg (8.12 ± 11.05 vs. 34.12 ± 54.31; P < 0.001) and Tg ratio (7.81 ± 8.98 vs. 20.01 ± 19.84; P < 0.001) were significantly higher in group 2. On the other hand, gender, tumor (T) stage, lymph node (N) stage, size, multiplicity or bilaterality of primary tumor, dose of 131I, serum TgAb and thyroid-stimulating hormone (TSH) level (before or after RAI therapy) were not significantly different in the two groups. Variables with P values of < 0.25 by univariate analysis were subjected to multivariate analysis, which showed post-ablation Tg (OR 1.060, 95 % CI = 1.028-1.092; P < 0.001) and Tg ratio (OR 1.059, 95 % CI = 1.028-1.092; P = 0.001) were significantly higher in group 2. CONCLUSION: Serum Tg level after RAI therapy was significantly higher in patients with midline uptake on RxWBS, compared with patients without midline uptake on RxWBS. Further investigations are needed to reveal the correlation between serum Tg elevation and clinical outcome according to the presence of midline uptake.

Giant Cell Tumor with Secondary Aneurysmal Bone Cyst Shows Heterogeneous Metabolic Pattern on 18F-FDG PET/CT: A Case Report.

Giant cell tumor (GCT) is a generally benign bone tumor accounting for approximately 5 % of all primary bone neoplasms. Cystic components in GCTs that indicate secondary aneurysmal bone cysts (ABCs) are reported in 14 % of GCTs. Although both of them have been described separately in previous reports that may show considerable fluorodeoxyglucose (FDG) uptake despite their benign nature, the findings of GCT with secondary ABC on 18F-FDG positron emission tomography/computed tomography (PET/CT) have not been well-known. We report a case of GCT with secondary ABC in a 26-year-old woman. 18F-FDG PET/CT revealed a heterogeneous hypermetabolic lesion in the left proximal femur with the maximum standardized uptake value of 4.7. The solid components of the tumor showed higher FDG uptake than the cystic components. These observations suggest that the ABC components in GCTs show heterogeneous metabolic patterns on 18F-FDG PET/CT.

Intratumoral Metabolic Heterogeneity for Prediction of Disease Progression After Concurrent Chemoradiotherapy in Patients with Inoperable Stage III Non-Small-Cell Lung Cancer.

PURPOSE: We evaluated the value of variable (18)F-FDG PET/CT parameters for the prediction of disease progression after concurrent chemoradiotherapy (CCRT) in patients with inoperable stage III non-small-cell lung cancer (NSCLC). METHODS: One hundred sixteen pretreatment FDG PET/CT scans of inoperable stage III NSCLC were retrospectively reviewed (stage IIIA: 51; stage IIIB: 65). The volume of interest was automatically drawn for each primary lung tumor, and PET parameters were assessed as follows: maximum standardized uptake value (SUVmax), metabolic tumor volume (MTV) using the boundaries presenting SUV intensity exceeding 3.0, and the area under the curve of the cumulative SUV-volume histograms (AUC-CSH), which is known to reflect the tumor heterogeneity. Progression-free survival (PFS), locoregional recurrence-free survival (LRFS), and distant metastasis-free survival (DMFS) were compared with each PET and clinical parameters by univariate and multivariate survival analysis. RESULTS: In the ROC analysis, the optimal cutoff values of SUVmax, MTV (cm(3)), and AUC-CSH for prediction of PFS were determined as 21.5, 27.7, and 4,800, respectively. In univariate analysis, PFS was statistically significantly reduced in those with AUC-CSH < 4,800 (p = 0.004). In multivariate analysis, AUC-CSH and SUVmax were statistically significant independent prognostic factors (HR 3.35, 95 % CI 1.79-6.28, p < 0.001; HR 0.25, 95 % CI 0.09-0.70, p = 0.008, respectively). Multivariate analysis showed that AUC-CSH was the most significant independent prognostic factor for LRFS and DMFS (HR 3.27, 95 % CI 1.54-6.94, p = 0.002; HR 2.79, 95 % CI 1.42-5.50, p = 0.003). CONCLUSIONS: Intratumoral metabolic heterogeneity of primary lung tumor in (18)F-FDG PET/CT can predict disease progression after CCRT in inoperable stage III NSCLC.

Stimulated Serum Thyroglobulin Level at the Time of First Dose of Radioactive Iodine Therapy Is the Most Predictive Factor for Therapeutic Failure in Patients With Papillary Thyroid Carcinoma.

PURPOSE: To investigate the clinical importance of serum thyroglobulin (Tg) levels just before high-dose I-131 ablation therapy (preablation Tg) for predicting therapeutic failure in patients with papillary thyroid carcinoma (PTC). METHODS: Patients with PTC (n = 132) undergoing total thyroidectomy followed by the first high-dose I-131 ablation therapy (HI-Rx) were included in this retrospective review. Just before HI-Rx, preablation Tg, anti-Tg antibody, and TSH were measured. The patients were followed up for a mean period of 7 months (range 6-23 months) by I-123 whole-body scans (f/u IWBS) and stimulated Tg (f/u Tg). Therapeutic failure was defined by positive f/u IWBS or f/u Tg >2 ng/ml. We classified patients into three groups according to the value of preablation Tg (group 1, <1 ng/ml; group 2, ≥1 and <10 ng/ml; group 3, ≥10 ng/ml) and compared clinical variables to therapeutic response. RESULTS: Therapeutic failure was noted in 39 patients (29.5 %). On univariate analysis, T stage, tumor size, and preablation Tg were the statistically significant factors that could predict therapeutic failure. After multivariate analysis, preablation Tg was the only independent predictor of therapeutic failure (P < 0.001). The therapeutic failure rate was significantly increased as the preablation Tg level increased (11.3 %, 33.3 %, and 87.5 % in groups 1, 2, and 3, respectively; P < 0.001). Individuals with preablation Tg levels ≥10 ng/ml had 25.5 times greater chance of therapeutic failure than those with levels <10 ng/ml (95 % CI = 5.43-119.60; P < 0.001). CONCLUSIONS: A high preablation Tg level is the most significant predictor of therapeutic failure at the time of first HI-Rx in patients with PTC.