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Background and Aims: In clinical trials, salvage therapy including Sofosbuvir/Velpatasvir/Voxilaprevir (SOF/VEL/VOX) achieved an SVR12 rate of 96% in NS5A-experienced participants. An extended access program (EAP) for SOF/VEL/VOX was sponsored by Gilead Pharmaceuticals for Australian patients with advanced liver disease or prior liver transplantation (LT), who had relapsed. We determined the real-world efficacy and safety of this regimen in this cohort with advanced liver disease. Method(s): In this Australian multicentre EAP study, adult patients with chronic HCV infection genotype(GT)1-6 with prior failure on a DAA NS5A inhibitor containing regimen were eligible to access 12 weeks of SOF/VEL/VOX 400 mg/100 mg/100 mg. Other eligibility criteria included compensated liver disease with at least one of the following i) Child Pugh (CP)A cirrhosis with clinically significant portal hypertension or platelets <100 x 109/L, ii) prior LT, or iii) severe extra hepatic manifestations. Hospitals with >1 EAP recipient were invited to participate. Clinical characteristics and treatment outcomes were recorded at baseline, end-of-treatment (EOT) and SVR12. HCV NS5A RAS testing was performed prior to retreatment, where serum was available. The primary outcomes was SVR12 rate. Result(s): To date, baseline data is available for 89 patients from 24 participating hospitals. 85 have reached end-of-treatment (EOT), and 80 have SVR12 data available for analysis. Four patients have incomplete data due to early discontinuation of therapy (n = 2), loss to follow up (n = 1) and death (n = 1). 74 (83%) were male and the median age was 53. GT3 was the most common (n = 64), then GT1a (n = 18), GT1b (n = 4), GT6 (n = 2) and GT4 (n = 1). The median platelet count was 127. 68 (75%) had cirrhosis and of these, 35 (51%) had portal hypertension. 14 (16%) had prior hepatoma and 15 (17%) prior LT. All participants were NS5A-experienced and 8 (9%) had received?>1 DAA course (1-4). Three participants were

Melanocytic pigmentation occurs in multiple sites in the lower female genital tract, but is rare within benign cysts of the vulva. We report 3 patients with multiloculated cystic lesions of the vulvar vestibule exhibiting prominent melanocytic pigmentation. The current cases differ from a previous report of melanosis in a Bartholin gland cyst in that the population of melanocytes occupies the acinar structures of the gland, rather than a squamous-lined surface. A similar cell population is demonstrated by immunoperoxidase methods in a fourth patient's nonpigmented gland, suggesting that melanin production may arise in a native, rather than metaplastic, cell population.Copyright ©2021International Society of Gynecological Pathologists.

Poliosis is a localized patch of depigmented grey or white hair. It may be caused by drugs, systemic conditions or as an immune mediated response to melanoma and even naevi. We describe an unusual presentation of new onset poliosis developing in 76 year old man surrounding two new blue pigmented lesions on the vertex of scalp that had initially been elevated. Initial punch biopsies of both lesions demonstrated pigment incontinence, macrophages without presence of melanocytes. An excisional biopsy was recommended for definitive histopathology which showed similar changes raising the suspicion for regressed melanoma. Wide local excision was recommended with resultant histopathology demonstrating in situ melanoma. This case highlights that regressed melanoma needs to be considered as a differential diagnosis in individuals presenting with poliosis and suspicious features such as pigmented lesions. Multiple biopsies or wider excision may be required to reach a diagnosis and provide appropriate treatment.

Introduction: Lower gastrointestinal haemorrhage (LGIH) is a challenging phenomenon in a comorbid, elderly population. CT mesenteric angiography (CTMA) allows localisation of the site of haemorrhage, and provides a target for interventional techniques, but the intermittent nature of LGIH makes it challenging to reliably demonstrate extravasation. This study aimed to identify objective factors that may predict scan outcomes. Method(s): In this retrospective cohort study, all patients undergoing CTMA for LGIH at Monash Health from January 2011 to December 2019 (n = 854) were included. Baseline patient characteristics included age, bowel resection/endoscopic intervention within the past 14 days, known bowel malignancy, anticoagulant/antiplatelet use, duration of symptoms, vital signs, transfusion requirements in the past 24 h and investigation results (recent haemoglobin levels, platelet count, international normalised ratio and creatinine levels). Univariate analysis was performed, and significant factors were entered into a multivariate model. Result(s): The final multivariate model was statistically significant (P < 0.001) and consisted of bowel resection/endoscopic intervention within the past 14 days (OR = 2.15), use of antiplatelet agents (OR = 2.03), blood transfusion requirement greater than 3 units per 24 h (OR = 1.79), systolic blood pressure less than 100 mmHg (OR = 1.56) and heart rate greater than 100 beats per minute (OR = 1.52). Conclusion(s): The factors identified above are objective, independently associated with positive scan outcomes, readily available to radiologists and are useful for more judicious patient selection.Copyright © 2021 The Royal Australian and New Zealand College of Radiologists

Background: DNA methylation is a potential biological mechanism through which residential greenness affects health, but little is known about its association with greenness and whether the association could be modified by genetic background. We aimed to evaluate the association between surrounding greenness and genome-wide DNA methylation and potential gene-greenness interaction effects on DNA methylation. Method(s): We measured blood-derived DNA methylation using the HumanMethylation450 BeadChip array (Illumina) for 479 Australian women, including 66 monozygotic, 66 dizygotic twin pairs, and 215 sisters of these twins. Surrounding greenness was represented by Normalized Difference Vegetation Index (NDVI) and Enhanced Vegetation Index (EVI) within 300, 500, 1000 or 2000 m surrounding participants' home addresses. For each cytosine-guanine dinucleotide (CpG), the associations between its methylation level and NDVI or EVI were evaluated by generalized estimating equations, after adjusting for age, education, marital status, area-level socioeconomic status, smoking behavior, cell-type proportions, and familial clustering. We used comb-p and DMRcate to identify significant differentially methylated regions (DMRs). For each significant CpG, we evaluated the interaction effects of greenness and single-nucleotide polymorphisms (SNPs) within +/-1 Mb window on its methylation level. Result(s): We found associations between surrounding greenness and blood DNA methylation for one CpG (cg04720477, mapped to the promoter region of CNP gene) with false discovery rate [FDR] < 0.05, and for another 9 CpGs with 0.05 <= FDR < 0.10. For two of these CpGs, we found 33 SNPs significantly (FDR < 0.05) modified the greenness-methylation association. There were 35 significant DMRs related to surrounding greenness that were identified by both comb-p (Sidak p-value < 0.01) and DMRcate (FDR < 0.01). Those CpGs and DMRs were mapped to genes related to many human diseases, such as mental health

Background and Aim: People with serious mental illness have high rates of bloodborne viruses (BBVs) but often have poor access to medical care. We sought to define the burden of hepatitis B virus (HBV), hepatitis C virus (HCV), and HIV in a long-stay inpatient cohort with serious mental illness. Emphasis was placed on elucidating BBV prevalence, risk factors, and access to care. Method(s): We prospectively approached all patients (n = 46) admitted to a long-stay mental health admission unit at a tertiary referring hospital over a 2-week period and invited each person to complete a BBV risk factor survey, opt-out BBV screening, and liver assessment using transient elastography and other non-invasive liver stiffness assays. This was part of a quality assurance initiative. Result(s): Baseline demographics showed a young cohort (median age, 39.5 years), with a male preponderance (71.7%). The primary psychiatric diagnoses were schizophrenia and schizoaffective disorders (95.7%). Almost two-thirds (63%) had a secondary diagnosis of substance-misuse disorders. Serology was obtained for 31 patients (67.4%). Of those screened for HBV, 15 (53.3%) were unvaccinated, four (13.3%) had previous HBV exposure, and 10 (33.3%) were vaccinated. No cases of HBV viremia were identified. HCV serology was positive (HCV antibody positive) in 11 patients (36.7%). Each had a history of intravenous drug use. Importantly, seven patients (23.3%) had successfully been treated for HCV and one (3.3%) had active viremia. The remaining cases likely reflected spontaneous clearance. To assess disease severity, transient elastography and the aspartate aminotransferase to platelet ratio index (APRI) were used as validated markers of liver fibrosis. APRI showed higher rates of fibrosis in the HCV antibody positive cohort, but this was not statistically significant compared with the HCV-naive cohort. Liver stiffness measurement conducted in the HCV antibody positive study group was similar and showed two

Objectives/Introduction: Obstructive sleep disordered breathing is highly prevalent in children with Down syndrome, largely attributed to the complex nature of the upper airway anatomical impairment in this population. However, the poor response to standard treatments targeting this anatomical impairment suggests the possible involvement of other non-anatomical factors such as neuromotor control of the upper airway muscles, arousal threshold and ventilatory control stability. Of these, ventilatory control instability (i.e. high loop gain) is of particular interest as recently developed techniques allows its estimation using standard polysomnography. Loop gain appears to have important clinical utility in adults with obstructive sleep apnoea, particularly in predicting individual responses to treatment. Accordingly, we aimed to investigate the role of ventilatory control instability in children with Down syndrome and sleep disordered breathing. Method(s): Children (3-18 years) with Down syndrome and sleep disordered breathing (n = 14) were compared with typically developing children (n = 14) matched for age, gender and sleep disordered breathing severity. All children underwent overnight polysomnography. Spontaneous sighs were identified and a 180-second analysis window (60s pre-sigh to 120s post-sigh) containing flow measurements and oxygen saturation were created. Loop gain, a measure of the sensitivity of the negative feedback loop that controls ventilation, was estimated by fitting a mathematical model of ventilatory control to the post-sigh ventilatory pattern. Result(s): Loop gain was significantly higher in children with Down syndrome compared to matched typically developing children (mean loop gain +/- standard deviation: 0.42 +/- 0.12 vs 0.31 +/- 0.11; p = 0.039). While children with Down syndrome also had significantly lower average oxygen saturation associated within each analysis window compared to typically developing children (96.9 +/- 1.3% vs 98.0 +

Gene expression profiling can be used for predicting survival in multiple myeloma (MM) and identifying patients who will benefit from particular types of therapy. Some germline single nucleotide polymorphisms (SNPs) act as expression quantitative trait loci (eQTLs) showing strong associations with gene expression levels. We performed an association study to test whether eQTLs of genes reported to be associated with prognosis of MM patients are directly associated with measures of adverse outcome. Using the genotype-tissue expression portal, we identified a total of 16 candidate genes with at least one eQTL SNP associated with their expression with P < 10-7 either in EBV-transformed B-lymphocytes or whole blood. We genotyped the resulting 22 SNPs in 1327 MM cases from the International Multiple Myeloma rESEarch (IMMEnSE) consortium and examined their association with overall survival (OS) and progression-free survival (PFS), adjusting for age, sex, country of origin and disease stage. Three polymorphisms in two genes (TBRG4-rs1992292, TBRG4-rs2287535 and ENTPD1-rs2153913) showed associations with OS at P <.05, with the former two also associated with PFS. The associations of two polymorphisms in TBRG4 with OS were replicated in 1277 MM cases from the International Lymphoma Epidemiology (InterLymph) Consortium. A meta-analysis of the data from IMMEnSE and InterLymph (2579 cases) showed that TBRG4-rs1992292 is associated with OS (hazard ratio = 1.14, 95% confidence interval 1.04-1.26, P =.007). In conclusion, we found biologically a plausible association between a SNP in TBRG4 and OS of MM patients.Copyright © 2021 The Authors. International Journal of Cancer published by John Wiley & Sons Ltd on behalf of Union for International Cancer Control.

Objective The study aimed to assess the association of nucleated red blood cells (NRBC), a surrogate of intrauterine hypoxia, and elevated pulmonic vascular resistance (E-PVR) and oxygen requirement after minimally invasive surfactant therapy (MIST). Study Design Retrospective study of a cohort of preterm neonates that received MIST in a single unit. Results NRBC were measured in 65 of 75 (87%) neonates administered MIST during the period. In total, 22 of 65 (34%) infants had pre-MIST echocardiography (ECHO). Neonates with elevated NRBC (predefined as >5 x 10 9 /L, n = 16) required higher post-MIST fraction of inspired oxygen (FiO 2) than neonates with normal NRBC (<1 x 10 9 /L, n = 17; FiO 2 = 0.31 +/- 0.10 and 0.24 +/- 0.04, respectively, p = 0.02). NRBC correlated positively with % of time in right to left ductal shunt (r = 0.51, p = 0.052) and inversely with right ventricular stroke volume (r = -0.55, p = 0.031) and time to peak velocity to right ventricular ejection time ratio (r = -0.62, p < 0.001). Conclusion Elevated NRBC are associated with elevated FiO 2 after MIST and elevated E-PVR. Intrauterine hypoxia may impact postnatal circulatory adaptations and oxygen requirement. Key Points Post-MIST FiO2 requirements are significantly higher in infants with elevated NRBC. NRBC correlates positively with elevated PVR in neonates requiring. Intrauterine hypoxia may play a role in postnatal circulatory adaptations in neonates with RDS.Copyright © 2021. Thieme. All rights reserved.

Human amnion epithelial cells (hAECs) exert potent antifibrotic and anti-inflammatory effects when transplanted into preclinical models of tissue fibrosis. These effects are mediated in part via the secretion of soluble factors by hAECs which modulate signaling pathways and affect cell types involved in inflammation and fibrosis. Based on these reports, we hypothesized that these soluble factors may also support liver regeneration during chronic liver injury. To test this, we characterized the effect of both hAECs and hAEC-conditioned medium (CM) on liver repair in a mouse model of carbon tetrachloride (CCl4)-induced fibrosis. Liver repair was assessed by liver fibrosis, hepatocyte proliferation, and the liver progenitor cell (LPC) response. We found that the administration of hAECs or hAEC-CM reduced liver injury and fibrosis, sustained hepatocyte proliferation, and reduced LPC numbers during chronic liver injury. Additionally, we undertook in vitro studies to document both the cell-cell and paracrine-mediated effects of hAECs on LPCs by investigating the effects of co-culturing the LPCs and hAECs and hAEC-CM on LPCs. We found little change in LPCs co-cultured with hAECs. In contrast, hAEC-CM enhances LPC proliferation and differentiation. These findings suggest that paracrine factors secreted by hAECs enhance liver repair by reducing fibrosis while promoting regeneration during chronic liver injury.Copyright © The Author(s) 2020.

We present a case of late symptom onset of COVID-19 infection 72 days after initial diagnosis in an immunocompromised 53-year-old man. SARS-CoV-2 was cultured from his sputum sample at this time, and genomic sequencing suggested reinfection was unlikely. After receipt of convalescent plasma, SARS-CoV-2 became undetectable by PCR 111 days after diagnosis, although SARS-CoV-2 antibodies remained not detectable. This case posed difficult public health management issues in a low prevalence COVID-19 setting as the person required extended home isolation given his prolonged SARS-CoV-2 PCR detection.Copyright © 2021 by The Society for Healthcare Epidemiology of America. All rights reserved.

Aim: To examine prescriber characteristics and reasoning for selection of stroke prevention therapy for dialysis patients with atrial fibrillation (AF). Background(s): Warfarin therapy in dialysis patients with AF remains controversial especially for primary stoke prevention. Method(s): Utilising scanned medical records, dialysis patients with AF at Monash Health were identified. Patient demographics, therapeutic agent for AF, prescriber characteristics and reasoning were then assessed. Result(s): Of 575 dialysis patients, 100 (17.4%) were identified as having AF. Of those, 31 patients received warfarin, 46 received antiplatelet therapy, 21 did not receive stroke prevention therapy and 2 received other forms of stroke prevention therapy (clexane non-dialysis days, apixaban). Of the 31 patients on warfarin, 15 were placed on warfarin for primary prevention, 13 for secondary prevention and 3 were unknown. The AF cohort was generally older compared to the cohort without AF (71.9 vs 61.2 years, p < 0.001). Demographically, patient subgroups on warfarin, antiplatelet agents and no stroke prevention therapy were similar. 16 of 31 patients were commenced on warfarin by cardiology, predominantly for primary stroke prevention (81%). Neurology commenced warfarin in 9 of 31 patients, in all instances for secondary stroke prevention (100%). 16 patients were previously on warfarin. Most cases of warfarin cessation were instigated by nephrology (63%) with major reasons including conflicting evidence regarding warfarin use in dialysis patients, bleeding complications, bleed risk greater than stroke risk, and dialysis commencement. Conclusion(s): Further research is required to provide an evidence-based approach to primary stroke prevention therapy in dialysis patients with AF. Presently, with limited evidence and differences in clinical practice between the various specialties, a multidisciplinary approach is necessary for the individualized management of AF in dialysis patients.

Many current-generation biomedical implants are fabricated from the Ti-6Al-4V alloy because it has many attractive properties, such as low density and biocompatibility. However, the elastic modulus of this alloy is much larger than that of the surrounding bone, leading to bone resorption and, eventually, implant failure. In the present study, we synthesized and performed a detailed analysis of a novel low elastic modulus Ti-based alloy (Ti-28Nb-5Zr-2Ta-2Sn (TNZTS alloy)) using a variety of methods, including scanning electron microscopy, transmission electron microscopy, X-ray diffraction, and tensile test. Additionally, the in vitro biocompatibility of the TNZTS alloy was evaluated using SCP-1, SaOs-2, and THP-1 cell lines and primary human osteoblasts. Compared to Ti-6Al-4V, the elastic modulus of TNZTS alloy was significantly lower, while measures of its in vitro biocompatibility are comparable. O2 plasma treatment of the surface of the alloy significantly increased its hydrophilicity and, hence, its in vitro biocompatibility. TNZTS alloy specimens did not induce the release of cytokines by macrophages, indicating that such scaffolds would not trigger inflammatory responses. The present results suggest that the TNZTS alloy may have potential as an alternative to Ti-6Al-4V. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Background: Monoclonal antibodies (mAbs) have proved to be a valuable tool for the treatment of different cancer types. However, clinical use of an increasing number of mAbs, have also highlighted limitations with monotherapy for cancers, in particular for such with more complex mechanisms, requiring action on additional molecules or pathways, or for cancers quickly acquiring resistance following monotherapy. An example for the latter is the mAb trastuzumab, FDA approved for treatment of metastatic gastric carcinoma. To circumvent this, researchers have reported synergistic, anti-proliferative effects by combination targeting of HER2 and EGFR by trastuzumab and the EGFR-targeting mAb Cetuximab overcoming trastuzumab resistance. Methods: Maintaining the proven functionality of trastuzumab, we have designed bi-specific antibody molecules, called AffiMabs, by fusing an EGFR-targeting Affibody molecule to trastuzumab’s heavy or light chains. Having confirmed binding to EGFR and Her2 and cytotoxicity of our AffiMabs, we analyzed apoptosis rate, receptor surface levels, phosphorylation levels of receptors and associated signaling pathways as well as differentially expressed genes on transcriptome level with the aim to elucidate the mode of action of our AffiMabs. Results: The AffiMabs are able to simultaneously bind HER2 and EGFR and show increased cytotoxic effect compared to the original trastuzumab therapeutic molecule and, more importantly, even to the combination of trastuzumab and EGFR-targeting Affibody molecule. Analyzing the mode of action, we could show that bi-specific AffiMabs lead to reduced surface receptor levels and a downregulation of cell cycle associated genes on transcriptome level. Conclusion: Our study shows that transcriptome analysis can be used to validate the choice of receptor targets and guide the design of novel multi-specific molecules. The inherent modularity of the AffiMab format renders it readily applicable to other receptor targets., QC 20220118

QC 20211116

Immunotherapy, especially anti-PD-1, is becoming a pillar of modern muscle-invasive bladder cancer (MIBC) treatment. However, the objective response rates (ORR) are relatively low due to the lack of precise biomarkers to select patients. Herein, the molecular subtype, tumor mutation burden (TMB), and CD8+ T cells were calculated by the gene expression and mutation profiles of MIBC patients. MIBC immunotypes were constructed using clustering analysis based on tumor mutation burden, CD8+ T cells, and molecular subtypes. Mutated genes, enriched functional KEGG pathways and GO terms, and co-expressed network-specific hub genes have been identified. We demonstrated that ORR of immunotype A patients identified by molecular subtype, CD8+ T cells, and TMB is about 36% predictable. PIK3CA, RB1, FGFR3, KMT2C, MACF1, RYR2, and EP300 are differentially mutated among three immunotypes. Pathways such as ECM-receptor interaction, PI3K-Akt signaling pathway, and TGF-beta signaling pathway are top-ranked in enrichment analysis. Low expression of ACTA2 was associated with the MIBC survival benefit. The current study constructs a model that could identify suitable MIBC patients for immunotherapy, and it is an important step forward to the personalized treatment of bladder cancers. © 2021, The Author(s).

Objective: We study activation of T helper 17 (Th17) and regulatory T (Treg) cells and induction of apoptosis in cells from patients with systemic lupus erythematosus (SLE) compared with controls and effects of atorvastatin and its simulated interactions with other compounds. Methods: Mononuclear cells from 10 patients with SLE and 10 controls were cultured in conditions that induce Th17 and/or Treg cell polarization and/or apoptosis and were studied by FACScan. Gene expression was determined by quantitative real-time reverse transcription–polymerase chain reaction. Cytokines in plasma were determined by enzyme-linked immunosorbent assay. The Search Tool for Interactions of Chemicals (STITCH) was used to retrieve information regarding the binding properties of atorvastatin. Results: Among patients with SLE, the proportion of Th17 (CD4+IL17+) cells was higher compared with controls after activation, with Th17 or Treg polarizing cytokines, phorbol myristate acetate, and ionomycin. In contrast, Treg cells (CD4+CD25+CD127dim/−) frequencies were lower. CD95 stimulation induced relatively more apoptosis in Treg cells and less in Th17 cells, as compared with controls. Addition of atorvastatin normalized Th17/Treg cell balance and apoptosis induction. Accordingly, the ratio of RORC/FoxP3 decreased in patients with SLE. Interleukin 17 and interleukin 6 (IL-6) levels were increased in patients with SLE. Atorvastatin interacted strongly with C-reactive protein (CRP) and also significantly with IL-6. Conclusion: There is a higher proportion of Th17 cells and a lower proportion of Treg cells in patients with SLE after activation. Th17 cells were more resistant than Treg cells to CD95-induced apoptosis in SLE. Atorvastatin normalized these effects. Our findings reveal a novel mechanism behind the imbalance of Th17/Treg cells with implications for treatment in SLE. We determine for the first time simulated interaction between atorvastatin, CRP, and IL-6, implying a novel role of a, QC 20230308

The development of multifunctional nanoscale systems that can mediate efficient tumor targeting, together with high cellular internalization, is crucial for the diagnosis of glioma. The combination of imaging agents into one platform provides dual imaging and allows further surface modification with targeting ligands for specific glioma detection. Herein, transferrin (Tf)-decorated niosomes with integrated magnetic iron oxide nanoparticles (MIONs) and quantum dots (QDs) were formulated (PEGNIO/QDs/MIONs/Tf) for efficient imaging of glioma, supported by magnetic and active targeting. Transmission electron microscopy confirmed the complete co-encapsulation of MIONs and QDs in the niosomes. Flow cytometry analysis demonstrated enhanced cellular uptake of the niosomal formulation by glioma cells. In vitro imaging studies showed that PEG-NIO/QDs/MIONs/Tf produces an obvious negative-contrast enhancement effect on glioma cells by magnetic resonance imaging (MRI) and also improved fluorescence intensity under fluorescence microscopy. This novel platform represents the first niosome-based system which combines magnetic nanoparticles and QDs, and has application potential in dual-targeted imaging of glioma. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

By accentuating drug efficacy and impeding resistance mechanisms, combinatorial, multi-agent therapies have emerged as key approaches in the treatment of complex diseases, most notably cancer. Using high-throughput drug screens, we uncovered distinct metabolic vulnerabilities and thereby identified drug combinations synergistically causing a starvation-like lethal catabolic response in tumor cells from different cancer entities. Domperidone, a dopamine receptor antagonist, as well as several tricyclic antidepressants (TCAs), including imipramine, induced cancer cell death in combination with the mitochondrial uncoupler niclosamide ethanolamine (NEN) through activation of the integrated stress response pathway and the catabolic CLEAR network. Using transcriptome and metabolome analyses, we characterized a combinatorial response, mainly driven by the transcription factors CHOP and TFE3, which resulted in cell death through enhanced pyrimidine catabolism as well as reduced pyrimidine synthesis. Remarkably, the drug combinations sensitized human organoid cultures to the standard-of-care chemotherapy paclitaxel. Thus, our combinatorial approach could be clinically implemented into established treatment regimen, which would be further facilitated by the advantages of drug repurposing. © 2021 The Authors. Published under the terms of the CC BY 4.0 license

Current diagnostic standards for lymphoproliferative disorders include multiple tests for detection of clonal immunoglobulin (IG) and/or T-cell receptor (TCR) rearrangements, translocations, copy-number alterations (CNAs), and somatic mutations. The EuroClonality-NGS DNA Capture (EuroClonality-NDC) assay was designed as an integrated tool to characterize these alterations by capturing IGH switch regions along with variable, diversity, and joining genes of all IG and TCR loci in addition to clinically relevant genes for CNA and mutation analysis. Diagnostic performance against standard-of-care clinical testing was assessed in a cohort of 280 B-and T-cell malignancies from 10 European laboratories, including 88 formalin-fixed paraffin-embedded samples and 21 reactive lesions. DNA samples were subjected to the EuroClonality-NDC protocol in 7 EuroClonality-NGS laboratories and analyzed using a bespoke bioinformatic pipeline. The EuroClonality-NDC assay detected B-cell clonality in 191 (97%) of 197 B-cell malignancies and T-cell clonality in 71 (97%) of 73 T-cell malignancies. Limit of detection (LOD) for IG/TCR rearrangements was established at 5% using cell line blends. Chromosomal translocations were detected in 145 (95%) of 152 cases known to be positive. CNAs were validated for immunogenetic and oncogenetic regions, highlighting their novel role in confirming clonality in somatically hypermutated cases. Single-nucleotide variant LOD was determined as 4% allele frequency, and an orthogonal validation using 32 samples resulted in 98% concordance. The EuroClonality-NDC assay is a robust tool providing a single end-To-end workflow for simultaneous detection of B-and T-cell clonality, translocations, CNAs, and sequence variants.

For millions of years, endogenous retroelements have remained transcriptionally silent within mammalian genomes by epigenetic mechanisms. Modern anticancer therapies targeting the epigenetic machinery awaken retroelement expression, inducing antiviral responses that eliminate tumors through mechanisms not completely understood. Here, we find that massive binding of epigenetically activated retroelements by RIG-I and MDA5 viral sensors promotes ATP hydrolysis and depletes intracellular energy, driving tumor killing independently of immune signaling. Energy depletion boosts compensatory ATP production by switching glycolysis to mitochondrial oxidative phosphorylation, thereby reversing the Warburg effect. However, hyperfunctional succinate dehydrogenase in mitochondrial electron transport chain generates excessive oxidative stress that unleashes RIP1-mediated necroptosis. To maintain ATP generation, hyperactive mitochondrial membrane blocks intrinsic apoptosis by increasing BCL2 dependency. Accordingly, drugs targeting BCL2 family proteins and epigenetic inhibitors yield synergistic responses in multiple cancer types. Thus, epigenetic therapy kills cancer cells by rewiring mitochondrial metabolism upon retroelement activation, which primes mitochondria to apoptosis by BH3-mimetics. SIGNIFICANCE: The state of viral mimicry induced by epigenetic therapies in cancer cells remodels mitochondrial metabolism and drives caspase-independent tumor cell death, which sensitizes to BCL2 inhibitor drugs. This novel mechanism underlies clinical efficacy of hypomethylating agents and venetoclax in acute myeloid leukemia, suggesting similar combination therapies for other incurable cancers.

There is little information on the dietary modulation of thrombosis-related risk factors such as platelet count. We aimed to assess the effects of Mediterranean diet (MedDiet) on platelet count and related outcomes in an older population at high cardiovascular risk. In participants of the PREDIMED (PREvencion con DIeta MEDiterranea) study, we assessed whether an intervention with a MedDiet enriched with extra-virgin olive oil or nuts, relative to a low-fat control diet, modulated platelet count (n = 4189), the risk of developing thrombocytosis and thrombocytopenia (n = 3086), and the association between these alterations and all-cause mortality (median follow-up time: 3.0 years). Although platelet count increased over time (+0.98 center dot 10(9) units/L center dot year [95% confidence interval: 0.12; 1.84]), MedDiet interventions moderated this increase, particularly in individuals with near-high baseline count (both MedDiets combined: -3.20 center dot 10(9) units/L center dot year [-5.81; -0.59]). Thrombocytopenia incidence was lower in the MedDiet interventions (incidence rates: 2.23% in control diet, 0.91% in MedDiets combined; hazard ratio: 0.44 [0.23; 0.83]). Finally, thrombocytopenia was associated with a higher risk of all-cause mortality (hazard ratio: 4.71 [2.69; 8.24]), but this relationship was attenuated in those allocated to MedDiet (p-interaction = 0.018). In brief, MedDiet maintained platelet counts within a healthy range and attenuated platelet-related mortality in older adults at high cardiovascular risk.

Simple Summary The main problem encountered in the management of prostate cancer (PCa) is the inability to distinguish slow-growing indolent tumors from aggressive tumors. It is therefore important to explore non-invasive assays for the early detection of this aggressive subtype, when it can still be treated effectively. The presence of the TWEAK cytokine in biofluids of the PCa microenvironment might drive the secretion of extracellular vesicles (EVs) containing exo-oncomicroRNAs capable of modifying the tumor microenvironment. These exo-oncomicroRNAs are potentially useful as PCa biomarkers. We identified 2 exo-oncomiRNAs isolated from semen EVs by the action of TWEAK in the tumor microenvironment and, we determined their usefulness as biomarkers of PCa prognostic. We also established, for the first time, that TWEAK modulates potential exo-oncomiRNA targets, both tightly linked to cancer progression. In conclusion, our study shows that semen detection of TWEAK-regulated exo-oncomiRNAs can improve PCa prognosis, opening new avenues for diagnosis and treatment. Liquid biopsy-based biomarkers, including microRNAs packaged within extracellular vesicles, are promising tools for patient management. The cytokine tumor necrosis factor-like weak inducer of apoptosis (TWEAK) is related to PCa progression and is found in the semen of patients with PCa. TWEAK can induce the transfer of exo-oncomiRNAs from tumor cells to body fluids, and this process might have utility in non-invasive PCa prognosis. We investigated TWEAK-regulated exo-microRNAs in semen and in post-digital rectal examination urine from patients with different degrees of PCa aggressiveness. We first identified 14 exo-oncomiRNAs regulated by TWEAK in PCa cells in vitro, and subsequently validated those using liquid

The pandemic caused by the SARS-CoV-2 infection affects many aspects of public health knowledge, science, and practice around the world. Several studies have shown that SARS-CoV-2 RNA in plasma seems to be associated with a worse prognosis of COVID-19. In the present study, we investigated plasma and buffy RNA in patients with COVID-19 to determine its prognostic value. A prospective study was carried out in patients hospitalized for COVID-19, in which RNA was analyzed in plasma and the buffy coat. Morphological and immunohistochemical studies were used to detect the presence of SARS-CoV-2 in the buffy coat. In COVID-19 patients, the obtained RNA concentration in plasma was 448.3 +/- 31.30 ng/mL. Of all the patients with positive plasma tests for SARS-CoV-2, 46.15% died from COVID-19. In four cases, tests revealed that SARS-CoV-2 was present in the buffy coat. Abnormal morphology of monocytes, lymphocytes and neutrophils was found. An immunohistochemical study showed positivity in mononuclear cells and platelets. Our results suggest that SARS-CoV-2 is present in the plasma. This facilitates viral dissemination and migration to specific organs, where SARS-CoV-2 infects target cells by binding to their receptors. In our study, the presence of plasma SARS-CoV-2 RNA was correlated with worse prognoses.

Our understanding of the interplay between human adipose tissue and the immune system is limited. The mesothelium, an immunologically active structure, emerged as a source of visceral adipose tissue. After investigating the mesothelial properties of human visceral and subcutaneous adipose tissue and their progenitors, we explored whether the dysfunctional obese and Crohn's disease environments influence the mesothelial/mesenchymal properties of their adipocyte precursors, as well as their ability to mount an immune response. Using a tandem transcriptomic/proteomic approach, we evaluated the mesothelial and mesenchymal expression profiles in adipose tissue, both in subjects covering a wide range of body-mass indexes and in Crohn's disease patients. We also isolated adipose tissue precursors (adipose-derived stem cells, ASCs) to assess their mesothelial/mesenchymal properties, as well as their antigen-presenting features. Human visceral tissue presented a mesothelial phenotype not detected in the subcutaneous fat. Only ASCs from mesenteric adipose tissue, named creeping fat, had a significantly higher expression of the hallmark mesothelial genes mesothelin (MSLN) and Wilms' tumor suppressor gene 1 (WT1), supporting a mesothelial nature of these cells. Both lean and Crohn's disease visceral ASCs expressed equivalent surface percentages of the antigen-presenting molecules human leucocyte antigen-DR isotype (HLA-DR) and CD86. However, lean-derived ASCs were predominantly HLA-DR (dim), whereas in Crohn's disease, the HLA-DR (bright) subpopulation was increased 3.2-fold. Importantly, the mesothelial-enriched Crohn's disease precursors activated CD4(+) T-lymphocytes. Our study evidences a mesothelial signature in the creeping fat of Crohn's disease patients and its progenitor c

Aim: To investigate DDR1 methylation in the brains of bipolar disorder (BD) patients and its association with DDR1 mRNA levels and comethylation with myelin genes. Materials & methods: Genome-wide profiling of DNA methylation (Infinium MethylationEPIC BeadChip) corrected for glial composition and DDR1 gene expression analysis in the occipital cortices of individuals with BD (n = 15) and healthy controls (n = 15) were conducted. Results: DDR1 5-methylcytosine levels were increased and directly associated with DDR1b mRNA expression in the brains of BD patients. We also observed that DDR1 was comethylated with a group of myelin genes. Conclusion: DDR1 is hypermethylated in BD brain tissue and is associated with isoform expression. Additionally, DDR1 comethylation with myelin genes supports the role of this receptor in myelination.