Coffee and Microbiota: A Narrative Review.

Coffee is one of the most widely consumed beverages in the world, which has important repercussions on the health of the individual, mainly because of certain compounds it contains. Coffee consumption exerts significant influences on the entire body, including the gastrointestinal tract, where a central role is played by the gut microbiota. Dysbiosis in the gut microbiota is implicated in the occurrence of numerous diseases, and knowledge of the microbiota has proven to be of fundamental importance for the development of new therapeutic strategies. In this narrative review, we thoroughly investigated the link between coffee consumption and its effects on the gut microbiota and the ensuing consequences on human health. We have selected the most significant articles published on this very interesting link, with the aim of elucidating the latest evidence about the relationship between coffee consumption, its repercussions on the composition of the gut microbiota, and human health. Based on the various studies carried out in both humans and animal models, it has emerged that coffee consumption is associated with changes in the gut microbiota, although further research is needed to understand more about this link and the repercussions for the whole organism.

The role of SARS-COV-2 infection in promoting abnormal immune response and sepsis: A comparison between SARS-COV-2-related sepsis and sepsis from other causes.

Background: COVID-19 caused by SARS-CoV-2 virus is characterized by respiratory compromise and immune system involvement, even leading to serious disorders, such as cytokine storm. Methods: We then conducted a literature review on the topic of sepsis and covid-19, and in parallel conducted an experimental study on the histological finding of patients who died from SARS-Covid 19 infection and a control group. Results: Sepsis associated with covid-19 infection has some similarities and differences from that from other causes. Conclusion: In this paper the complex interplay between the 2 disorders was discussed, focusing on the similarities and on the effect that one could have on the other. A preliminary experimental section that demonstrates the multisystemic involvement in subjects who die from SARS-CoV-2 is also proposed.

Activity-dependent regulation of the BAX/BCL-2 pathway protects cortical neurons from apoptotic death during early development.

During early brain development, homeostatic removal of cortical neurons is crucial and requires multiple control mechanisms. We investigated in the cerebral cortex of mice whether the BAX/BCL-2 pathway, an important regulator of apoptosis, is part of this machinery and how electrical activity might serve as a set point of regulation. Activity is known to be a pro-survival factor; however, how this effect is translated into enhanced survival chances on a neuronal level is not fully understood. In this study, we show that caspase activity is highest at the neonatal stage, while developmental cell death peaks at the end of the first postnatal week. During the first postnatal week, upregulation of BAX is accompanied by downregulation of BCL-2 protein, resulting in a high BAX/BCL-2 ratio when neuronal death rates are high. In cultured neurons, pharmacological blockade of activity leads to an acute upregulation of Bax, while elevated activity results in a lasting increase of BCL-2 expression. Spontaneously active neurons not only exhibit lower Bax levels than inactive neurons but also show almost exclusively BCL-2 expression. Disinhibition of network activity prevents the death of neurons overexpressing activated CASP3. This neuroprotective effect is not the result of reduced caspase activity but is associated with a downregulation of the BAX/BCL-2 ratio. Notably, increasing neuronal activity has a similar, non-additive effect as the blockade of BAX. Conclusively, high electrical activity modulates BAX/BCL-2 expression and leads to higher tolerance to CASP3 activity, increases survival, and presumably promotes non-apoptotic CASP3 functions in developing neurons.

The Impact of Smoking on Microbiota: A Narrative Review.

Cigarette smoke is a classic risk factor for many diseases. The microbiota has been recently indicated as a new, major player in human health. Its deregulation-dysbiosis-is considered a new risk factor for several illnesses. Some studies highlight a cross-interaction between these two risk factors-smoke and dysbiosis-that may explain the pathogenesis of some diseases. We searched the keywords "smoking OR smoke AND microbiota" in the title of articles on PubMed®, UptoDate®, and Cochrane®. We included articles published in English over the last 25 years. We collected approximately 70 articles, grouped into four topics: oral cavity, airways, gut, and other organs. Smoke may impair microbiota homeostasis through the same harmful mechanisms exerted on the host cells. Surprisingly, dysbiosis and its consequences affect not only those organs that are in direct contact with the smoke, such as the oral cavity or the airways, but also involve distant organs, such as the gut, heart, vessels, and genitourinary tract. These observations yield a deeper insight into the mechanisms implicated in the pathogenesis of smoke-related diseases, suggesting a role of dysbiosis. We speculate that modulation of the microbiota may help prevent and treat some of these illnesses.

Gut Microbiota, LADA, and Type 1 Diabetes Mellitus: An Evolving Relationship.

There is much evidence confirming the crucial role played by the gut microbiota in modulating the immune system in the onset of autoimmune diseases. In this article, we focus on the relationship between alterations in the microbiome and the onset of diabetes mellitus type 1 and LADA, in light of the latest evidence. We will then look at both how the role of the gut microbiota appears to be increasingly crucial in the pathogenesis of these disorders and how this aspect may be instrumental in the development of new potential therapeutic strategies that modulate the gut microbiota, such as probiotics, prebiotics, and fecal microbiota transplantation.

Chronic running-wheel exercise from adolescence leads to increased anxiety and depression-like phenotypes in adulthood in rats: Effects on stress markers and interaction with BDNF Val66Met genotype.

Exercise has been shown to be beneficial in reducing symptoms of affective disorders and to increase the expression of brain-derived neurotrophic factor (BDNF). The BDNF Val66Met polymorphism is associated with reduced activity-dependent BDNF release and increased risk for anxiety and depression. Male and female Val66Met rats were given access to running wheels from 3 weeks of age and compared to sedentary controls. Anxiety- and depression-like behaviors were measured in adulthood using the elevated plus maze (EPM), open field (OF), and forced swim test (FST). Expression of BDNF and a number of stress-related genes, the glucocorticoid receptor (Nr3c1), serum/glucocorticoid-regulated kinase 1 (Sgk1), and FK506 binding protein 51 (Fkbp5) in the hippocampus were also measured. Rats given access to running wheels developed high levels of voluntary exercise, decreased open-arm time on the EPM and center-field time in the OF, reduced overall exploratory activity in the open field, and increased immobility time in the FST with no differences between genotypes. Chronic exercise induced a significant increase in Bdnf mRNA and BDNF protein levels in the hippocampus with some of these effects being genotype specific. Exercise decreased the expression of Nr3c1 and Sgk1, but increased the expression of Fkbp5. These results suggest that chronic running-wheel exercise from adolescence increased anxiety and depression-like phenotypes in adulthood, independent of BDNF Val66Met genotype. Further studies are required to confirm that increased indices of anxiety-like behavior are independent from reduced overall locomotor activity.

Gut Microbiota and Clostridium difficile: What We Know and the New Frontiers.

Our digestive system, particularly our intestines, harbors a vast amount of microorganisms, whose genetic makeup is referred to as the microbiome. Clostridium difficile is a spore-forming Gram-positive bacterium, which can cause an infection whose symptoms range from asymptomatic colonization to fearsome complications such as the onset of toxic megacolon. The relationship between gut microbiota and Clostridium difficile infection has been studied from different perspectives. One of the proposed strategies is to be able to specifically identify which types of microbiota alterations are most at risk for the onset of CDI. In this article, we understood once again how crucial the role of the human microbiota is in health and especially how crucial it becomes, in the case of its alteration, for the individual's disease. Clostridium difficile infection is an emblematic example of how a normal and physiological composition of the human microbiome can play a very important role in immune defense against such a fearsome disease.

When Autoimmunity 'DRESSes up': A Case after Certolizumab Therapy.

Drug Reaction with Eosinophilia and Systemic Symptoms (DRESS) is characterised by skin rash together with visceral organ involvement, lymphadenopathy, eosinophilia and atypical lymphocytosis. The syndrome is clinically heterogeneous, making diagnosis challenging. It has an annual incidence of 2 per 100,000 population and a mortality rate of 2-10%. We describe the first case of DRESS induced by certolizumab, a biologic disease-modifying antirheumatic drug (bioDMARD). LEARNING POINTS: Drug Reaction with Eosinophilia and Systemic Symptoms (DRESS) is an uncommon and under-reported syndrome.Its recognition is critical for treatment, especially in the emergency setting where most patients first present.In the case of unexplained fever, lymphadenopathy, cutaneous rash and characteristic laboratory findings (e.g., eosinophilia), after infectious causes have been ruled out, clinicians should always keep DRESS in mind and consider possible recent intake of a triggering drug.

Use of POCUS in Chest Pain and Dyspnea in Emergency Department: What Role Could It Have?

Chest pain and dyspnea are common symptoms in patients presenting to the emergency room (ER); oftentimes it is not possible to clearly identify the underlying cause, which may cause the patient to have to return to the ER. In other cases, while it is possible to identify the underlying cause, it is necessary to perform a large number of tests before being able to make a diagnosis. Over the last twenty years, emergency medicine physicians have had the possibility of using ultrasound to help them make and rule out diagnoses. Specific ultrasound tests have been designed to evaluate patients presenting with specific symptoms to ensure a fast, yet complete, evaluation. In this paper, we examine the role of ultrasound in helping physicians understand the etiology behind chest pain and dyspnea. We analyze the different diseases and disorders which may cause chest pain and dyspnea as symptoms and discuss the corresponding ultrasound findings.

Behavioral phenotyping of a rat model of the BDNF Val66Met polymorphism reveals selective impairment of fear memory.

The common brain-derived neurotrophic factor (BDNF) Val66Met polymorphism is associated with reduced activity-dependent BDNF release and increased risk for anxiety disorders and PTSD. Here we behaviorally phenotyped a novel Val66Met rat model with an equivalent valine to methionine substitution in the rat Bdnf gene (Val68Met). In a three-day fear conditioning protocol of fear learning and extinction, adult rats with the Met/Met genotype demonstrated impaired fear memory compared to Val/Met rats and Val/Val controls, with no genotype differences in fear learning or extinction. This deficit in fear memory occurred irrespective of the sex of the animals and was not seen in adolescence (4 weeks of age). There were no changes in open-field locomotor activity or anxiety measured in the elevated plus maze (EPM) nor in other types of memory measured using the novel-object recognition test or Y-maze. BDNF exon VI expression in the dorsal hippocampus was higher and BDNF protein level in the ventral hippocampus was lower in female Val/Met rats than female Val/Val rats, with no other genotype differences, including in total BDNF, BDNF long, or BDNF IV mRNA. These data suggest a specific role for the BDNF Met/Met genotype in fear memory in rats. Further studies are required to investigate gene-environment interactions in this novel animal model.

Is Dual Mobility Total Hip Arthroplasty Surgery More Aggressive than Hemiarthroplasty when Treating Femoral Neck Fracture in the Elderly? A Multicentric Retrospective Study on 302 Hips.

INTRODUCTION: Bipolar hemiarthroplasty (BHA) and total hip arthroplasty (THA) are validated treatments for displaced femoral neck fractures (DFNFs). BHA seldomly needs conversion to THA, but the latter has higher dislocation rate in FNFs. Dual Mobility THA offers a reduced dislocation rate and eliminates the risk of conversion. This study looks for differences between BHA and DMTHA in terms of surgical time, blood loss and transfusion, dislocation rate, mortality, and thromboembolic events. MATERIAL AND METHODS: All patients were ≥75yo. Recorded data included use of anticoagulant/antiplatelet drugs, ASA, operative time, intra-operative complications, pre/post-operative hemoglobin values, transfusions, hospitalization time, DVT/PE, glomerular filtration rate, Charlson Comorbidity Index (CCI), dislocation at 60 days, and mortality at 30 days and 6 months. A secondary analysis compared the subgroups in different age range (75-85 and ≥ 86yo). RESULTS: In the cohort of 302 DFNF (93 BHA and 209 DMTHA) differences in mean age, CCI, and ASA score were significant. Once divided by age, the subgroups resulted comparable in terms of age and CCI, with no significant difference. A significant difference in surgical times showed DMTHA being an average 12 minutes longer than BHA. Significant was the ΔHB in the DMTHA subgroup which resulted lower compared to the BHA one. Difference in mean number of post-operative transfusion were not statistically significant. CONCLUSIONS: From our data, DMTHA did not lead to an increase in mortality, morbidity, bleeding, or dislocation rate when compared to BHA and could be considered as treatment of choice for DFNFs especially in healthy and active patients.

Microbiota and Myopericarditis: The New Frontier in the Car-Diological Field to Prevent or Treat Inflammatory Cardiomyo-Pathies in COVID-19 Outbreak.

Myopericarditis is an inflammatory heart condition involving the pericardium and myocardium. It can lead to heart failure, dilated cardiomyopathy, arrhythmia and sudden death. Its pathogenesis is mainly mediated by viral infections but also can be induced by bacterial infections, toxic substances and immune mediated disorders. All these conditions can produce severe inflammation and myocardial injury, often associated with a poor prognosis. The specific roles of these different pathogens (in particular viruses), the interaction with the host, the interplay with gut microbiota, and the immune system responses to them are still not completely clear and under investigation. Interestingly, some research has demonstrated the contribution of the gut microbiota, and its related metabolites (some of which can mimic the cardiac myosin), in cardiac inflammation and in the progression of this disease. They can stimulate a continuous and inadequate immune response, with a subsequent myocardial inflammatory damage. The aim of our review is to investigate the role of gut microbiota in myopericarditis, especially for the cardiovascular implications of COVID-19 viral infection, based on the idea that the modulation of gut microbiota can be a new frontier in the cardiological field to prevent or treat inflammatory cardiomyopathies.

CDK4/6 Inhibitor as a Novel Therapeutic Approach for Advanced Bladder Cancer Independently of RB1 Status.

PURPOSE: Bladder cancer is a clinical and social problem due to its high incidence and recurrence rates. It frequently appears in elderly patients showing other medical comorbidities that hamper the use of standard chemotherapy. We evaluated the activity of CDK4/6 inhibitor as a new therapy for patients unfit for cisplatin (CDDP). EXPERIMENTAL DESIGN: Bladder cancer cell lines were tested for in vitro sensitivity to CDK4/6 inhibition. A novel metastatic bladder cancer mouse model was developed and used to test its in vivo activity. RESULTS: Cell lines tested were sensitive to CDK4/6 inhibition, independent on RB1 gene status. Transcriptome analyses and knockdown experiments revealed a major role for FOXM1 in this response. CDK4/6 inhibition resulted in reduced FOXM1 phosphorylation in vitro and in vivo and showed synergy with CDDP, allowing a significant tumor regression. FOXM1 exerted important oncogenic roles in bladder cancer. CONCLUSIONS: CDK4/6 inhibitors, alone or in combination, are a novel therapeutic strategy for patients with advanced bladder cancer previously classified as unfit for current treatment options.

Kidney transplant from uncontrolled donation after circulatory death donors maintained by nECMO has long-term outcomes comparable to standard criteria donation after brain death.

Uncontrolled donation after circulatory death (uDCD) increases organ availability for kidney transplant (KT) with short-term outcomes similar to those obtained from donation after brain death (DBD) donors. However, heterogeneous results in the long term have been reported. We compared 10-year outcomes between 237 KT recipients from uDCD donors maintained by normothermic extracorporeal membrane oxygenation (nECMO) and 237 patients undergoing KT from standard criteria DBD donors during the same period at our institution. We further analyzed risk factors for death-censored graft survival in the uDCD group. Delayed graft function (DGF) was more common in the uDCD group (73.4% vs 46.4%; P < .01), although glomerular filtration rates at the end of follow-up were similar in the 2 groups. uDCD and DBD groups had similar rates for 10-year death-censored graft (82.1% vs 80.4%; P = .623) and recipient survival (86.2% vs 87.6%; P = .454). Donor age >50 years was associated with graft loss in the uDCD group (hazard ratio: 1.91; P = .058), whereas the occurrence of DGF showed no significant effect. uDCD KT under nECMO support resulted in similar graft function and long-term outcomes compared with KT from standard criteria DBD donors. Increased donor age could negatively affect graft survival after uDCD donation.

47. Síndrome platipnea ortodeoxia por foramen oval permeable luego de implante de stent en aorta torácica

Historia clínica: varón de 70 años con antecedente de ACV sin secuelas neurológicas. Niega otros factores de riesgo CV y presenta un examen físico normal. En valoración por su ACV se solicitan estudios complementarios. Pruebas complementarias: Doppler de vasos de cuello sin ateromatosis significativas. ETT: FEVI 45%, hipoquinesia inferior y biauriculomegalia leve. Rx de tórax: mediastino ensanchado a nivel superior. TAC de aorta: aneurisma sacular en arco aórtico. Evolución clínica: se decide reparación endovascular mediante implante percutáneo de un stent multicapa diversor de flujo (40 mm de diámetro por 38 cm de largo) desde la aorta ascendente a la aorta abdominal. Luego de finalizado dicho procedimiento instala rápidamente insuficiencia respiratoria severa que requiere ventilación invasiva e ingreso a CTI por 48 horas. Se realiza nueva TAC en la que no se identifican complicaciones vinculadas al procedimiento, descartando oclusiones arteriales, tromboembolismo pulmonar y neumotórax. En la evolución presenta mejoría paulatina desde el punto de vista respiratorio por lo que se extuba e ingresa a la unidad cardiológica. Se constata saturación de oxígeno en decúbito dorsal de 97% con máscara de flujo libre 5 l/min y semisentado de 87% (confirmado por gasometría). Se sospecha síndrome platipnea ortodeoxia, solicitándose ETE que muestra gran aneurisma del septum interauricular con marcada excursión hacia ambas aurículas, separación de ambas láminas del septum interauricular con flujo derecha izquierda, pasaje masivo de burbujas de derecha a izquierda potenciada con maniobra de Valsalva y válvula de Eustaquio prominente. Evoluciona con mejoría gradual y diminución de la diferencia de oxemia posicional. Diagnóstico: se plantea síndrome platipnea ortodeoxia secundario a endoprótesis aórtica. Discusión: el síndrome de platipnea ortodeoxia es una causa poco habitual de disnea e hipoxemia. En este paciente la patogenia probablemente se deba a un fenómeno mecánico al momento del implante del stent aórtico que determinó una modificación en la posición de las estructuras cardíacas. En un paciente con foramen oval permeable no diagnosticado previamente y con una válvula de Eustaquio prominente, se favoreció el direccionamiento de flujo de derecha a izquierda. La compresión de la AD en bipedestación facilitaría el pasaje dinámico de flujo entre las aurículas aumentando el shunt intracardíaco.

BMP4 Induces M2 Macrophage Polarization and Favors Tumor Progression in Bladder Cancer.

Purpose: Bladder cancer is a current clinical and social problem. At diagnosis, most patients present with nonmuscle-invasive tumors, characterized by a high recurrence rate, which could progress to muscle-invasive disease and metastasis. Bone morphogenetic protein (BMP)-dependent signaling arising from stromal bladder tissue mediates urothelial homeostasis by promoting urothelial cell differentiation. However, the possible role of BMP ligands in bladder cancer is still unclear.Experimental Design: Tumor and normal tissue from 68 patients with urothelial cancer were prospectively collected and analyzed for expression of BMP and macrophage markers. The mechanism of action was assessed in vitro by experiments with bladder cancer cell lines and peripheral blood monocyte-derived macrophages.Results: We observed BMP4 expression is associated and favored type II macrophage differentiation. In vitro experiments showed that both recombinant BMP4 and BMP4-containing conditioned media from bladder cancer cell lines favored monocyte/macrophage polarization toward M2 phenotype macrophages, as shown by the expression and secretion of IL10. Using a series of human bladder cancer patient samples, we also observed increased expression of BMP4 in advanced and undifferentiated tumors in close correlation with epithelial-mesenchymal transition (EMT). However, the p-Smad 1,5,8 staining in tumors showing EMT signs was reduced, due to the increased miR-21 expression leading to reduced BMPR2 expression.Conclusions: These findings suggest that BMP4 secretion by bladder cancer cells provides the M2 signal necessary for a protumoral immune environment. In addition, the repression of BMPR2 by miR-21 makes the tumor cells refractory to the prodifferentiating actions mediated by BMP ligands, favoring tumor growth. Clin Cancer Res; 23(23); 7388-99. ©2017 AACR.

Treatment of caval vein thrombosis associated with renal tumors. / Tratamiento de la trombosis de vena cava inferior asociada a los tumores renales.

INTRODUCTION: Renal carcinoma represents 3% of all solid tumors and is associated with renal or inferior caval vein (IVC) thrombosis between 2-10% of patients, extending to right atrial in 1% of cases. METHODS: This is a retrospective study that comprises 5 patients who underwent nephrectomy and thrombectomy by laparotomy because of renal tumor with IVC thrombosis level iii. RESULTS: Four patients were males and one was female, and the mean age was 57,2 years (range: 32-72). Most important clinical findings were hematuria, weight loss, weakness, anorexia, and pulmonary embolism. Diagnostic confirmation was performed by CT scanner. Metastatic disease was diagnosed before surgery in 3 patients. Suprahepatic caval vein and hepatic hilium (Pringle's maneouver) were clamped in 4 patients, and ligation of infrarrenal caval vein was carry out in one patient. Five patients developed mild complications (Clavien I/II). No patient died and the mean hospital stay was 8,6 days. All patients were treated with chemotherapy, and 3 died because distant metastasis, but 2 are alive, without recurrence, at 5 and 60 months, respectively. CONCLUSIONS: Nephrectomy and thrombectomy in renal tumors with caval thrombosis can be curative in absence of metastasis or, at less, can increase survival or quality of live. Then these patients must be treated in liver transplant units because major surgical and anesthesiologic expertise. Adjuvant treatment with tyrosin kinase inhibitors must be validate in the future with wider experiences.

Tratamiento de la trombosis de vena cava inferior asociada a los tumores renales / Treatment of caval vein thrombosis associated with renal tumors

INTRODUCCIÓN: El carcinoma renal representa el 3% de los tumores sólidos y se asocia a trombosis de la vena renal o vena cava inferior (VCI) en el 2-10% de los pacientes; se extiende hasta la aurícula derecha en el 1% de los casos. MÉTODOS: Estudio retrospectivo de una serie de 5 enfermos intervenidos por tumor renal con trombosis tumoral de VCI de nivel III, tratados con nefrectomía y trombectomía por laparotomía. RESULTADOS: Cuatro de los pacientes eran hombres y uno era mujer, con una edad media de 57,2 años (rango: 32-72). Como clínica predominó la hematuria, síndrome constitucional y tromboembolia pulmonar. La confirmación diagnóstica fue por TAC. En 3 pacientes se detectaron metástasis antes de la cirugía. Se realizó Pringle y pinzamiento de vena cava suprahepática en 4 pacientes y ligadura de VCI infrarrenal en uno. Complicaciones leves (Clavien I/II) se presentaron en 5 pacientes. La mortalidad fue nula y la estancia hospitalaria media fue de 8,6 días. Todos los pacientes se trataron con quimioterapia; fallecieron 3 por metástasis a distancia, permanecen 2 vivos, sin recidiva, a los 5 y 60 meses. Supervivencia media: 26,6 meses. CONCLUSIONES: La nefrectomía y la trombectomía en tumores renales con trombosis de cava pueden ser curativas en ausencia de metástasis o, al menos, pueden aumentar la supervivencia y mejorar la calidad de vida. Para ello estos pacientes deberían tratarse en unidades de trasplante hepático por su mayor experiencia quirúrgica y anestésica. El tratamiento adyuvante con inhibidores de la tirosina cinasa debe validarse en el futuro con experiencias más amplias

INTRODUCTION: Renal carcinoma represents 3% of all solid tumors and is associated with renal or inferior caval vein (IVC) thrombosis between 2-10% of patients, extending to right atrial in 1% of cases. METHODS: This is a retrospective study that comprises 5 patients who underwent nephrectomy and thrombectomy by laparotomy because of renal tumor with IVC thrombosis level III. RESULTS: Four patients were males and one was female, and the mean age was 57,2 years (range: 32-72). Most important clinical findings were hematuria, weight loss, weakness, anorexia, and pulmonary embolism. Diagnostic confirmation was performed by CT scanner. Metastatic disease was diagnosed before surgery in 3 patients. Suprahepatic caval vein and hepatic hilium (Pringle's maneouver) were clamped in 4 patients, and ligation of infrarrenal caval vein was carry out in one patient. Five patients developed mild complications (Clavien I/II). No patient died and the mean hospital stay was 8,6 days. All patients were treated with chemotherapy, and 3 died because distant metastasis, but 2 are alive, without recurrence, at 5 and 60 months, respectively. CONCLUSIONS: Nephrectomy and thrombectomy in renal tumors with caval thrombosis can be curative in absence of metastasis or, at less, can increase survival or quality of live. Then these patients must be treated in liver transplant units because major surgical and anesthesiologic expertise. Adjuvant treatment with tyrosin kinase inhibitors must be validate in the future with wider experiences

UroMark-a urinary biomarker assay for the detection of bladder cancer.

BACKGROUND: Bladder cancer (BC) is one of the most common cancers in the western world and ranks as the most expensive to manage, due to the need for cystoscopic examination. BC shows frequent changes in DNA methylation, and several studies have shown the potential utility of urinary biomarkers by detecting epigenetic alterations in voided urine. The aim of this study is to develop a targeted bisulfite next-generation sequencing assay to diagnose BC from urine with high sensitivity and specificity. RESULTS: We defined a 150 CpG loci biomarker panel from a cohort of 86 muscle-invasive bladder cancers and 30 normal urothelium. Based on this panel, we developed the UroMark assay, a next-generation bisulphite sequencing assay and analysis pipeline for the detection of bladder cancer from urinary sediment DNA. The 150 loci UroMark assay was validated in an independent cohort (n = 274, non-cancer (n = 167) and bladder cancer (n = 107)) voided urine samples with an AUC of 97%. The UroMark classifier sensitivity of 98%, specificity of 97% and NPV of 97% for the detection of primary BC was compared to non-BC urine. CONCLUSIONS: Epigenetic urinary biomarkers for detection of BC have the potential to revolutionise the management of this disease. In this proof of concept study, we show the development and utility of a novel high-throughput, next-generation sequencing-based biomarker for the detection of BC-specific epigenetic alterations in urine.