Multi-domain neurocognitive impairment following definitive intensity-modulated radiotherapy for nasopharyngeal cancer: A cross-sectional study.

INTRODUCTION: Neurocognitive impairment from inadvertent brain irradiation is common following intensity-modulated radiotherapy (IMRT) for nasopharyngeal carcinoma (NPC). This study aimed to determine the prevalence, pattern, and radiation dose-toxicity relationship of this late complication. MATERIALS AND METHODS: We undertook a cross-sectional study of 190 post-IMRT NPC survivors. Neurocognitive function was screened using the Montreal Cognitive Assessment-Hong Kong (HK-MoCA). Detailed assessments of eight distinct neurocognitive domains were conducted: intellectual capacity (WAIS-IV), attention span (Digit Span and Visual Spatial Span), visual memory (Visual Reproduction Span), verbal memory (Auditory Verbal Learning Test), processing speed (Color Trail Test), executive function (Stroop Test), motor dexterity (Grooved Pegboard Test) and language ability (Verbal Fluency Test). The mean percentiles and Z-scores were compared with normative population data. Associations between radiation dose and brain substructures were explored using multivariable logistic regression. RESULTS: The median post-IMRT interval was 7.0 years. The prevalence of impaired HK-MoCA was 25.3 % (48/190). Among the participants, 151 (79.4 %) exhibited impairments in at least one neurocognitive domain. The predominantly impaired domains included verbal memory (short-term: mean Z-score, -0.56, p < 0.001; long-term: mean Z-score, -0.70, p < 0.001), processing speed (basic: mean Z-score, -1.04, p < 0.001; advanced: mean Z-score, -0.38, p < 0.001), executive function (mean Z-score, -1.90, p < 0.001), and motor dexterity (dominant hand: mean Z-score, -0.97, p < 0.001). Radiation dose to the whole brain, hippocampus, and temporal lobe was associated with impairments in executive function, verbal memory, processing speed, and motor dexterity. CONCLUSIONS: Neurocognitive impairment is prevalent and profound in post-IMRT NPC survivors. Cognitive assessment and rehabilitation should be considered part of survivorship care.

Diffuse C-Cells Hyperplasia Is the Source of False Positive Calcitonin Measurement in FNA Washout Fluids of Thyroid Nodules: A Rational Clinical Approach to Avoiding Unnecessary Surgery.

PURPOSE: The FNA-CT is useful for the diagnosis of MTC. The aim of this study was to evaluate the performance of FNA-CT in TNs coexisting with CCH. METHODS: This study retrospectively reviewed the records of 11 patients with TNs submitted to thyroidectomy on the basis of elevated basal and/or stimulated serum CT values, which at histology were not confirmed to be MTC. The results obtained in this group were compared with those of a previously reported group of histologically proven MTC patients submitted to an identical presurgical evaluation. All patients, negative for known mutations in the RET proto-oncogene, were preoperatively submitted to neck ultrasound, FNA-cytology, and FNA-CT. RESULTS: Approximately 6 of 11 patients showed increased (>36 ng/mL, as established in previous studies not involving patients with CCH) FNA-CT. All these patients showed diffuse CCH at histology in the thyroid lobe submitted to FNA; 5 of them were benign at histology, while only one was malignant (papillary thyroid carcinoma, PTC). The remaining 5 of 11 patients had low FNA-CT (<36 ng/mL), and all of them showed only focal CCH in the lobe submitted to FNA; three of them were malignant (2 PTC, 1 follicular carcinoma), while two were benign. CONCLUSIONS: Employing the currently proposed cut-off values, false-positive FNA-CT results may be observed in benign/malignant TNs with coexisting diffuse CCH. FNA-CT must therefore be cautiously used in the diagnostic approach for patients with TNs and a slightly increased basal or stimulated serum CT concentration in order to avoid unnecessary surgery.

FT-6876, a Potent and Selective Inhibitor of CBP/p300, is Active in Preclinical Models of Androgen Receptor-Positive Breast Cancer.

BACKGROUND: Patients with triple-negative breast cancer (TNBC) expressing the androgen receptor (AR) respond poorly to neoadjuvant chemotherapy, although AR antagonists have shown promising clinical activity, suggesting these tumors are AR-dependent. cAMP responsive element binding protein (CREB)-binding protein (CBP) and p300 are transcriptional co-activators for the AR, a key driver of AR+ breast and prostate cancer, and may provide a novel therapeutic target in AR+ TNBC. OBJECTIVES: The aim of this study was to determine the therapeutic potential of FT-6876, a new CBP/p300 bromodomain inhibitor, in breast cancer models with a range of AR levels in vitro and in vivo. METHODS: Effects of FT-6876 on the CBP/p300 pathway were determined by combining chromatin immunoprecipitation (ChIP) with precision run-on sequencing (PRO-seq) complemented with H3K27 acetylation (Ac) and transcriptional profiling. The antiproliferative effect of FT-6876 was also measured in vitro and in vivo. RESULTS: We describe the discovery of FT-6876, a potent and selective CBP/p300 bromodomain inhibitor. The combination of ChIP and PRO-seq confirmed the reduction in H3K27Ac at specific promoter sites concurrent with a decrease in CBP/p300 on the chromatin and a reduction in nascent RNA and enhancer RNA. This was associated with a time- and concentration-dependent reduction in H3K37Ac associated with a decrease in AR and estrogen receptor (ER) target gene expression. This led to a time-dependent growth inhibition in AR+ models, correlated with AR expression. Tumor growth inhibition was also observed in AR+ tumor models of TNBC and ER+ breast cancer subtypes with consistent pharmacokinetics and pharmacodynamics. CONCLUSION: Our findings demonstrate FT-6876 as a promising new CBP/p300 bromodomain inhibitor, with efficacy in preclinical models of AR+ breast cancer.

Lifestyle Changes Reduced Estimated White Matter Hyperintensities Based on Retinal Image Analysis.

This study evaluates if there is an association between lifestyle changes and the risk of small vessel disease (SVD) as measured by cerebral white matter hyperintensities (WMH) estimated by the automatic retinal image analysis (ARIA) method. We recruited 274 individuals into a community cohort study. Subjects were assessed at baseline and annually with the Health-Promoting Lifestyle Profile II Questionnaire (HPLP-II) and underwent a simple physical assessment. Retinal images were taken using a non-mydriatic digital fundus camera to evaluate the level of WMH estimated by ARIA (ARIA-WMH) to measure the risk of small vessel disease. We calculated the changes from baseline to one year for the six domains of HPLP-II and analysed the relationship with the ARIA-WMH change. A total of 193 (70%) participants completed both the HPLP-II and ARIA-WMH assessments. The mean age was 59.1 ± 9.4 years, and 76.2% (147) were women. HPLP-II was moderate (Baseline, 138.96 ± 20.93; One-year, 141.97 ± 21.85). We observed a significant difference in ARIA-WMH change between diabetes and non-diabetes subjects (0.03 vs. -0.008, respectively, p = 0.03). A multivariate analysis model showed a significant interaction between the health responsibility (HR) domain and diabetes (p = 0.005). For non-diabetes subgroups, those with improvement in the HR domain had significantly decreased in ARIA-WMH than those without HR improvement (-0.04 vs. 0.02, respectively, p = 0.003). The physical activity domain was negatively related to the change in ARIA-WMH (p = 0.02). In conclusion, this study confirms that there is a significant association between lifestyle changes and ARIA-WMH. Furthermore, increasing health responsibility for non-diabetes subjects reduces the risk of having severe white matter hyperintensities.

Digital solution for detection of undiagnosed diabetes using machine learning-based retinal image analysis.

INTRODUCTION: Undiagnosed diabetes is a global health issue. Previous studies have estimated that about 24.1%-75.1% of all diabetes cases are undiagnosed, leading to more diabetic complications and inducing huge healthcare costs. Many current methods for diabetes diagnosis rely on metabolic indices and are subject to considerable variability. In contrast, a digital approach based on retinal image represents a stable marker of overall glycemic status. RESEARCH DESIGN AND METHODS: Our study involves 2221 subjects for developing a classification model, with 945 subjects with diabetes and 1276 controls. The training data included 70% and the testing data 30% of the subjects. All subjects had their retinal images taken using a non-mydriatic fundus camera. Two separate data sets were used for external validation. The Hong Kong testing data contain 734 controls without diabetes and 660 subjects with diabetes, and the UK testing data have 1682 subjects with diabetes. RESULTS: The 10-fold cross-validation using the support vector machine approach has a sensitivity of 92% and a specificity of 96.2%. The separate testing data from Hong Kong provided a sensitivity of 99.5% and a specificity of 91.1%. For the UK testing data, the sensitivity is 98.0%. The accuracy of the Caucasian retinal images is comparable with that of the Asian data. It implies that the digital method can be applied globally. Those with diabetes complications in both Hong Kong and UK data have a higher probability of risk of diabetes compared with diabetes subjects without complications. CONCLUSIONS: A digital machine learning-based method to estimate the risk of diabetes based on retinal images has been developed and validated using both Asian and Caucasian data. Retinal image analysis is a fast, convenient, and non-invasive technique for community health applications. In addition, it is an ideal solution for undiagnosed diabetes prescreening.

Association between hashimoto thyroiditis and differentiated thyroid cancer: A single-center experience.

Hashimoto's thyroiditis is the most common cause of hypothyroidism in the iodine-sufficient areas of the world. Differentiated thyroid cancer is the most common thyroid cancer subtype, accounting for more than 95% of cases, and it is considered a tumor with a good prognosis, although a certain number of patients experience a poor clinical outcome. Hashimoto's thyroiditis has been found to coexist with differentiated thyroid cancer in surgical specimens, but the relationship between these two entities has not yet been clarified. Our study aims to analyze the relationship between these two diseases, highlighting the incidence of histological diagnosis of Hashimoto thyroiditis in differentiated thyroid cancer patients, and assess how this autoimmune disorder influences the risk of structural disease recurrence and recurrence rate.

Machine-learning method for localization of cerebral white matter hyperintensities in healthy adults based on retinal images.

Retinal vessels are known to be associated with various cardiovascular and cerebrovascular disease outcomes. Recent research has shown significant correlations between retinal characteristics and the presence of cerebral small vessel disease as measured by white matter hyperintensities from cerebral magnetic resonance imaging. Early detection of age-related white matter changes using retinal images is potentially helpful for population screening and allow early behavioural and lifestyle intervention. This study investigates the ability of the machine-learning method for the localization of brain white matter hyperintensities. All subjects were age 65 or above without any history of stroke and dementia and recruited from local community centres and community networks. Subjects with known retinal disease or disease influencing vessel structure in colour retina images were excluded. All subjects received MRI on the brain, and age-related white matter changes grading was determined from MRI as the primary endpoint. The presence of age-related white matter changes on each of the six brain regions was also studied. Retinal images were captured using a fundus camera, and the analysis was done based on a machine-learning approach. A total of 240 subjects are included in the study. The analysis of various brain regions included the left and right sides of frontal lobes, parietal-occipital lobes and basal ganglia. Our results suggested that data from both eyes are essential for detecting age-related white matter changes in the brain regions, but the retinal parameters useful for estimation of the probability of age-related white matter changes in each of the brain regions may differ for different locations. Using a classification and regression tree approach, we also found that at least three significant heterogeneous subgroups of subjects were identified to be essential for the localization of age-related white matter changes. Namely those with age-related white matter changes in the right frontal lobe, those without age-related white matter changes in the right frontal lobe but with age-related white matter changes in the left parietal-occipital lobe, and the rest of the subjects. Outcomes such as risks of severe grading of age-related white matter changes and the proportion of hypertension were significantly related to these subgroups. Our study showed that automatic retinal image analysis is a convenient and non-invasive screening tool for detecting age-related white matter changes and cerebral small vessel disease with good overall performance. The localization analysis for various brain regions shows that the classification models on each of the six brain regions can be done, and it opens up potential future clinical application.

Genome-wide binding analysis of 195 DNA binding proteins reveals "reservoir" promoters and human specific SVA-repeat family regulation.

A key aspect in defining cell state is the complex choreography of DNA binding events in a given cell type, which in turn establishes a cell-specific gene-expression program. Here we wanted to take a deep analysis of DNA binding events and transcriptional output of a single cell state (K562 cells). To this end we re-analyzed 195 DNA binding proteins contained in ENCODE data. We used standardized analysis pipelines, containerization, and literate programming with R Markdown for reproducibility and rigor. Our approach validated many findings from previous independent studies, underscoring the importance of ENCODE's goals in providing these reproducible data resources. We also had several new findings including: (i) 1,362 promoters, which we refer to as 'reservoirs,' that are defined by having up to 111 different DNA binding-proteins localized on one promoter, yet do not have any expression of steady-state RNA (ii) Reservoirs do not overlap super-enhancer annotations and distinct have distinct properties from super-enhancers. (iii) The human specific SVA repeat element may have been co-opted for enhancer regulation and is highly transcribed in PRO-seq and RNA-seq. Collectively, this study performed by the students of a CU Boulder computational biology class (BCHM 5631 -Spring 2020) demonstrates the value of reproducible findings and how resources like ENCODE that prioritize data standards can foster new findings with existing data in a didactic environment.

Metabolic Alteration in Plasma and Biopsies From Patients With IBD.

BACKGROUND: Inflammatory bowel diseases (IBD) are chronic inflammatory disorders of the gastrointestinal tract, with periods of latency alternating with phases of exacerbation, and include 2 forms: Crohn disease (CD) and ulcerative colitis (UC). Although the etiology of IBD is still unclear, the identification and understanding of pathophysiological mechanisms underlying IBD could reveal newly targeted intestinal alterations and determine therapeutic approaches. METHODS: In this study, by using gas chromatography-mass spectrometry, we characterized plasma and biopsies from the metabolomics profiles of patients with IBD compared with those of a control group. RESULTS: The results showed a different metabolomics profile between patients with CD (n = 50) and patients with UC (n = 82) compared with the control group (n = 51). Multivariate statistical analysis of the identified metabolites in CD and UC showed changes in energetic metabolism, and lactic acid and ornithine in particular were altered in both plasma and colon biopsies. Moreover, metabolic changes were evidenced between the normal ileum and colon tissues. These differences disappeared when we compared the inflamed ileum and colon tissues, suggesting a common metabolism. CONCLUSIONS: This study showed how the metabolomics profile could be a potential tool to identify intestinal alterations associated with IBD and may have application in precision medicine and for better defining the pathogenesis of the disease.

Parathyroid Carcinoma in the Setting of Tertiary Hyperparathyroidism: Case Report and Review of the Literature.

INTRODUCTION: Parathyroid carcinoma is one of the rarest cancers in normal population, and it is extremely uncommon in the setting of tertiary hyperparathyroidism. Indeed, only 24 cases have been reported in the literature. Presentation of the Case. We report the case of parathyroid carcinoma in a 51-year-old man, with a history of end-stage renal disease due to a horseshoe kidney treated with haemodialysis since 2013. He came to our attention due to an increase in calcium and parathyroid hormone serum levels. Neck ultrasound (US) showed a solid hypodense mass, probably the right inferior parathyroid gland, with an estimated size of 25 × 15 × 13 mm; the 99mTc-sestamibi SPECT/CT scan revealed a large radiotracer activity area in the right cervical region, compatible with a hyperfunctioning right inferior parathyroid gland. So, a tertiary hyperparathyroidism diagnosis was made. In April 2018, resection of three parathyroid glands was performed. Histopathological examination demonstrated the right inferior parathyroid gland specimen to be a parathyroid carcinoma, due to the presence of multiple, full-thickness, capsular infiltration foci, and a venous vascular invasion focus. Discussion. Diagnosis of parathyroid carcinoma in tertiary hyperparathyroidism is remarkably complex because of the lack of clinical diagnostic criteria and, in many cases, is made postoperatively at histopathological examination. CONCLUSION: To date, radical surgery represents the mainstay of treatment, with a five- and ten-year survival rates overall acceptable.

A machine learning approach for retinal images analysis as an objective screening method for children with autism spectrum disorder.

BACKGROUND: Autism spectrum disorder (ASD) is characterised by many of features including problem in social interactions, different ways of learning, some children showing a keen interest in specific subjects, inclination to routines, challenges in typical communication, and particular ways of processing sensory information. Early intervention and suitable supports for these children may make a significant contribution to their development. However, considerable difficulties have been encountered in the screening and diagnosis of ASD. The literature has indicated that certain retinal features are significantly associated with ASD. In this study, we investigated the use of machine learning approaches on retinal images to further enhance the classification accuracy. METHODS: Forty-six ASD participants were recruited from three special needs schools and 24 normal control were recruited from the community. Among them, 23 age-gender matched ASD and normal control participant-pairs were constructed for the primary analysis. All retinal images were captured using a nonmydriatic fundus camera. Automatic retinal image analysis (ARIA) methodology applying machine-learning technology was used to optimise the information of the retina to develop a classification model for ASD. The model's validity was then assessed using a 10-fold cross-validation approach to assess its validity. FINDINGS: The sensitivity and specificity were 95.7% (95% CI 76.0%, 99.8%) and 91.3% (95% CI 70.5%, 98.5%) respectively. The area under the ROC curve was 0.974 (95% CI 0.934, 1.000); however, it was noted that the specificity for female participants might not be as high as that for male participants. INTERPRETATION: Because ARIA is a fully automatic cloud-based algorithm and relies only on retinal images, it can be used as a risk assessment tool for ASD screening. Further diagnosis and confirmation can then be made by professionals, and potential treatment may be provided at a relatively early stage.

Association between obesity, common chronic diseases and health promoting lifestyle profiles in Hong Kong adults: a cross-sectional study.

BACKGROUND: Recent surveys revealed that the health status of many people from Hong Kong is far from ideal. Although non-communicable diseases are largely preventable, few relevant health promotion and disease prevention programs are available. Thus, we assessed the health indicators of Chinese adults in Hong Kong to investigate the relationship between obesity, common chronic diseases, and health-promoting lifestyle profiles to provide inspirations for decision makers in formulating targeted disease prevention and health management programs. METHODS: This is a secondary analysis of a data set of 270 community-dwelling Hong Kong adults who were within the eligible age range between 18 and 80 years without eye diseases that affect retinal photographs. The study exposure variable, health-promoting lifestyle profiles, was measured using the Health-Promoting Lifestyle Profile II (HPLP-II) questionnaire. The primary outcome variable, obesity, was defined using body mass index and waist-hip ratio. The secondary study outcome, estimated chronic diseases, including of anemia, chronic kidney disease, and cardiovascular disease, were estimated using automatic retinal image analysis from the retinal images. Data were analyzed using tests of proportion, the independent sample t-tests, Welch's t-test, and binary logistic regression models. RESULTS: All HPLP-II subscales had positive responses (≥ 2.5). Significant differences were noted between men and women in the health responsibility and nutrition subscales (Health Responsibility: p = 0.059; Nutrition: p = 0.067). Regression models revealed that nutrition (adjusted odds ratio [AOR] = 0.41; p = 0.017), physical activity (AOR = 0.50; p = 0.015), interpersonal relations (AOR = 2.14; p = 0.016), and stress management (AOR = 2.07; p 0.038) were associated with obesity; while spiritual growth (AOR = 0.24; p = 0.077) and interpersonal relations (AOR = 5.06; p 0.069) were associated with estimated chronic kidney disease. CONCLUSIONS: Improving health behaviors may control or alleviate the prevalence of obesity and chronic kidney disease. These findings could arouse concern about lifestyle behaviors and promote self-assessment of health-promoting lifestyles to the general public. The study also provided new insights into the relationship between the HPLP-II and other common chronic diseases that warrant further study.

Predictive Factors of Lymph Node Metastasis in Patients With Papillary Microcarcinoma of the Thyroid: Retrospective Analysis on 293 Cases.

Introduction: Papillary thyroid microcarcinoma (PTMC) is defined as a tumor with a larger diameter ≤ 1 cm and is considered having an indolent course and an excellent prognosis. Nevertheless, the incidence of lymph node metastasis in PTMC is not negligible, reaching up to 65% in some series. The aim of this study was to assess the incidence of lymph node metastasis in patients with PTMC and to evaluate predictive factors for lymph node metastasis. Methods: We included in this retrospective observational study patients who underwent thyroidectomy with pathological diagnosis of PTMC at our department from January 2003 to June 2019. Results: Two hundred ninety-three patients were included in the study. The incidence of lymph node metastasis was 13.7%. Multivariate analysis revealed as independent risk factors for lymph node metastasis age <45 years, nodule size ≥6 mm, tall cell variant of PTC, extrathyroidal extension, and angioinvasion. Conversely, autoimmune thyroiditis was found as a protective factor for lymph node metastasis. A subgroup of patients, with nodule size ≤ 5 mm, presented non-aggressive features. Conclusion: The incidence of lymph node metastasis in PTMC is considerable; the size of the tumor appears to be the most significant predictive factor for lymph node metastasis. The traditional cut-off value used for definition of microcarcinoma could be reconsidered to identify patients with an indolent course of the tumor, where active surveillance could be the appropriate treatment, and on the other hand, patients with potentially aggressive tumors requiring an adequate surgical intervention. Clinical Trial Registration: The trial was registered at ClinicalTrials.gov (ID: NCT04274829).

Predictive Factors of Recurrence in Patients with Differentiated Thyroid Carcinoma: A Retrospective Analysis on 579 Patients.

Differentiated thyroid carcinoma (DTC) is usually associated with a favorable prognosis. Nevertheless, up to 30% of patients present a local or distant recurrence. The aim of this study was to assess the incidence of recurrence after surgery for DTC and to identify predictive factors of recurrence. We included in this retrospective study 579 consecutive patients who underwent thyroidectomy for DTC from 2011 to 2016 at our institution. We observed biochemical or structural recurrent disease in 36 (6.2%) patients; five-year disease-free survival was 94.1%. On univariate analysis, male sex, histotype, lymph node yield, lymph node metastasis, extrathyroidal invasion and multicentricity were associated with significantly higher risk of recurrence, while microcarcinoma was correlated with significantly lower risk of recurrence. On multivariate analysis, only lymph node metastases (OR 4.724, p = 0.012) and microcarcinoma (OR 0.328, p = 0.034) were detected as independent predictive factors of recurrence. Postoperative management should be individualized and commensurate with the risk of recurrence: Patients with high-risk carcinoma should undergo strict follow-up and aggressive treatment. Furthermore, assessment of the risk should be repeated over time, considering individual response to therapy.

Retinal image analytics detects white matter hyperintensities in healthy adults.

Objective: We investigated whether an automatic retinal image analysis (ARIA) incorporating machine learning approach can identify asymptomatic older adults harboring high burden of white matter hyperintensities (WMH) using MRI as gold standard. Methods: In this cross-sectional study, we evaluated 180 community-dwelling, stroke-, and dementia-free healthy subjects and performed ARIA by acquiring a nonmydriatic retinal fundus image. The primary outcome was the diagnostic performance of ARIA in detecting significant WMH on MRI brain, defined as age-related white matter changes (ARWMC) grade ≥2. We analyzed both clinical variables and retinal characteristics using logistic regression analysis. We developed a machine learning network model with ARIA to estimate WMH and its classification. Results: All 180 subjects completed MRI and ARIA. The mean age was 70.3 ± 4.5 years, 70 (39%) were male. Risk factor profiles were: 106 (59%) hypertension, 31 (17%) diabetes, and 47 (26%) hyperlipidemia. Severe WMH (global ARWMC grade ≥2) was found in 56 (31%) subjects. The performance for detecting severe WMH with sensitivity (SN) 0.929 (95% CI from 0.819 to 0.977) and specificity (SP) 0.984 (95% CI from 0.937 to 0.997) was excellent. There was a good correlation between WMH volume (log-transformed) obtained from MRI versus those estimated from retinal images using ARIA with a correlation coefficient of 0.897 (95% CI from 0.864 to 0.922). Interpretation: We developed a robust algorithm to automatically evaluate retinal fundus image that can identify subjects with high WMH burden. Further community-based prospective studies should be performed for early screening of population at risk of cerebral small vessel disease.

Endothelial Cell-Specific Inactivation of TSPAN12 (Tetraspanin 12) Reveals Pathological Consequences of Barrier Defects in an Otherwise Intact Vasculature.

Objective- Blood-CNS (central nervous system) barrier defects are implicated in retinopathies, neurodegenerative diseases, stroke, and epilepsy, yet, the pathological mechanisms downstream of barrier defects remain incompletely understood. Blood-retina barrier (BRB) formation and retinal angiogenesis require ß-catenin signaling induced by the ligand norrin (NDP [Norrie disease protein]), the receptor FZD4 (frizzled 4), coreceptor LRP5 (low-density lipoprotein receptor-like protein 5), and the tetraspanin TSPAN12 (tetraspanin 12). Impaired NDP/FZD4 signaling causes familial exudative vitreoretinopathy, which may lead to blindness. This study seeked to define cell type-specific functions of TSPAN12 in the retina. Approach and Results- A loxP-flanked Tspan12 allele was generated and recombined in endothelial cells using a tamoxifen-inducible Cdh5-CreERT2 driver. Resulting phenotypes were documented using confocal microscopy. RNA-Seq, histopathologic analysis, and electroretinogram were performed on retinas of aged mice. We show that TSPAN12 functions in endothelial cells to promote vascular morphogenesis and BRB formation in developing mice and BRB maintenance in adult mice. Early loss of TSPAN12 in endothelial cells causes lack of intraretinal capillaries and increased VE-cadherin (CDH5 [cadherin5 aka VE-cadherin]) expression, consistent with premature vascular quiescence. Late loss of TSPAN12 strongly impairs BRB maintenance without affecting vascular morphogenesis, pericyte coverage, or perfusion. Long-term BRB defects are associated with immunoglobulin extravasation, complement deposition, cystoid edema, and impaired b-wave in electroretinograms. RNA-sequencing reveals transcriptional responses to the perturbation of the BRB, including genes involved in vascular basement membrane alterations in diabetic retinopathy. Conclusions- This study establishes mice with late endothelial cell-specific loss of Tspan12 as a model to study pathological consequences of BRB impairment in an otherwise intact vasculature.

Author Correction: Cross sectional evaluation of the gut-microbiome metabolome axis in an Italian cohort of IBD patients.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Cross sectional evaluation of the gut-microbiome metabolome axis in an Italian cohort of IBD patients.

Inflammatory bowel disease (IBD) is a chronic inflammatory disease of the gastrointestinal tract of uncertain origin, which includes ulcerative colitis (UC) and Crohn's disease (CD). The composition of gut microbiota may change in IBD affected individuals, but whether dysbiosis is the cause or the consequence of inflammatory processes in the intestinal tissue is still unclear. Here, the composition of the microbiota and the metabolites in stool of 183 subjects (82 UC, 50 CD, and 51 healthy controls) were determined. The metabolites content and the microbiological profiles were significantly different between IBD and healthy subjects. In the IBD group, Firmicutes, Proteobacteria, Verrucomicrobia, and Fusobacteria were significantly increased, whereas Bacteroidetes and Cyanobacteria were decreased. At genus level Escherichia, Faecalibacterium, Streptococcus, Sutterella and Veillonella were increased, whereas Bacteroides, Flavobacterium, and Oscillospira decreased. Various metabolites including biogenic amines, amino acids, lipids, were significantly increased in IBD, while others, such as two B group vitamins, were decreased in IBD compared to healthy subjects. This study underlines the potential role of an inter-omics approach in understanding the metabolic pathways involved in IBD. The combined evaluation of metabolites and fecal microbiome can be useful to discriminate between healthy subjects and patients with IBD.

Norrin-induced Frizzled4 endocytosis and endo-lysosomal trafficking control retinal angiogenesis and barrier function.

Angiogenesis and blood-brain barrier formation are required for normal central nervous system (CNS) function. Both processes are controlled by Wnt or Norrin (NDP) ligands, Frizzled (FZD) receptors, and ß-catenin-dependent signalling in vascular endothelial cells. In the retina, FZD4 and the ligand NDP are critical mediators of signalling and are mutated in familial exudative vitreoretinopathy. Here, we report that NDP is a potent trigger of FZD4 ubiquitination and induces internalization of the NDP receptor complex into the endo-lysosomal compartment. Inhibition of ubiquitinated cargo transport through the multivesicular body (MVB) pathway using a dominant negative ESCRT (endosomal sorting complexes required for transport) component VPS4 EQ strongly impairs NDP/FZD4 signalling in vitro and recapitulates CNS angiogenesis and blood-CNS-barrier defects caused by impaired vascular ß-catenin signalling in mice. These findings provide evidence for an important role of FZD4 endocytosis in NDP/FZD4 signalling and in CNS vascular biology and disease.

Estimation of clinical parameters of chronic kidney disease by exhaled breath full-scan mass spectrometry data and iterative PCA with intensity screening algorithm.

Breath mass spectrometry is a useful tool for identifying important compounds associated with health. However, there have been few studies that have explored human exhaled breath by full-scan mass spectrometry as a non-invasive method for medical diagnosis, which may be attributed to the difficulties resulting from multicollinearity and small sample sizes relative to a large number of product ions. In this study, breath samples from 54 chronic kidney disease patients were analyzed by selected ion flow tube mass spectrometry in the full-scan mode. With the signal intensities of product ions, we developed a novel and robust algorithm, iterative PCA with intensity screening (IPS), to build linear models for estimating important clinical parameters of chronic kidney disease. It has been shown that IPS provided good estimations in cross-validated samples, and furthermore the identified product ions could have direct medical relevance to the disease. The study demonstrated the potential of quantitative breath analysis using mass spectrometry for medical diagnosis, and the importance of applying appropriate statistical tools to unveil the rich information in this type of data.