Extracellular Vesicles from Human Cerebrospinal Fluid Are Effectively Separated by Sepharose CL-6B-Comparison of Four Gravity-Flow Size Exclusion Chromatography Methods.

Extracellular vesicles (EVs) are a versatile group of cell-secreted membranous nanoparticles present in body fluids. They have an exceptional diagnostic potential due to their molecular content matching the originating cells and accessibility from body fluids. However, methods for EV isolation are still in development, with size exclusion chromatography (SEC) emerging as a preferred method. Here we compared four types of SEC to isolate EVs from the CSF of patients with severe traumatic brain injury. A pool of nine CSF samples was separated by SEC columns packed with Sepharose CL-6B, Sephacryl S-400 or Superose 6PG and a ready-to-use qEV10/70 nm column. A total of 46 fractions were collected and analysed by slot-blot followed by Ponceau staining. Immunodetection was performed for albumin, EV markers CD9, CD81, and lipoprotein markers ApoE and ApoAI. The size and concentration of nanoparticles in fractions were determined by tunable resistive pulse sensing and EVs were visualised by transmission electron microscopy. We show that all four SEC techniques enabled separation of CSF into nanoparticle- and free protein-enriched fractions. Sepharose CL-6B resulted in a significantly higher number of separated EVs while lipoproteins were eluted together with free proteins. Our data indicate that Sepharose CL-6B is suitable for isolation of EVs from CSF and their separation from lipoproteins.

Acute Liver Failure as the Leading Manifestation of Spontaneous Tumour Lysis Syndrome in a Patient with NonHodgkin Lymphoma: Do Current Diagnostic Criteria of Tumour Lysis Syndrome Need Re-Evaluation?

Tumour lysis syndrome (TLS) is a group of pathophysiological processes caused by rapid degradation of tumour cells with subsequent release of intracellular contents into the extracellular space. It is characterized by the development of systemic metabolic disturbances with or without clinical manifestations. The process usually occurs in highly proliferative, large tumours after induction of cytotoxic therapy. Rarely, however, spontaneous TLS can develop, as well as signs of multiorgan failure triggered by an excessive metabolic load and sterile inflammation. The combination of the aforementioned is thus quite unique. Here, we present a 63-year-old male in which spontaneous TLS was accompanied with acute liver failure and delineated underlying nonHodgkin lymphoma. Initial laboratory findings included hyperkalaemia, hyperphosphataemia, hypocalcaemia, uraemia, and increased creatinine levels indicating the onset of TLS with acute kidney injury. Moreover, the patient showed signs of jaundice, coagulopathy, and hepatic encephalopathy. Development of TLS with multiorgan failure prompted rapid initiation of critical care management, including vigorous intravenous fluid therapy, allopurinol treatment, high flow continuous venovenous haemodiafiltration, and commencement of chemotherapy. The case highlights the possibility of TLS as a differential diagnosis in patients presenting with multiorgan failure and the importance of early detection of this potentially challenging and fatal diagnosis.

Erectile function in cardiovascular patients: its significance and a quick assessment using a visual-scale questionnaire.

PURPOSE: The aim of this study was to investigate the connection between erectile dysfunction (ED) and cardiovascular diseases and to test a novel visual-scale questionnaire (VEF) we propose for the assessment of erectile function. SUBJECTS AND METHODS: Erectile function was assessed in 170 male cardiovascular patients under the age of 70 by the use of several self-administered questionnaires: the International Index of Erectile Function-5 (IIEF-5); the Massachusetts Male Aging Study questionnaires (MMAS Sexual Activity Questionnaire and MMAS Single Question), and finally, VEF. RESULTS: Patients' mean age was 55.65 ± 9.97 y. The most common indications for hospitalization were coronary artery disease (CAD) (n = 82, 48%), and decompensated chronic heart failure (n = 30, 18%). The prevalence of ED as determined by IIEF-5 was 58% (n = 99). Patients with ED were on average 5.7 years older (P = 0.0001), had a higher frequency of diabetes (by 19%, P < 0.01), and a somewhat higher level of uric acid (by 72 µmol/l, P < 0.01). Results of the VEF correlated significantly with those of other questionnaires. Three different machine learning algorithms demonstrated a greater accuracy of VEF than IIEF-5 and MMAS Sexual Activity Questionnaire in predicting ED severity. CONCLUSION: ED is highly prevalent among cardiovascular patients. The Visual Scale Erectile Function questionnaire (VEF) is a simple and valid tool, suitable for quick screening of this condition.

Chronic granulomatous inflammation of the breast as a first clinical manifestation of primary sarcoidosis.

BACKGROUND: Sarcoidosis is an idiopathic multisystemic disease that affects young to middle aged adults, with higher incidence in women. Although it may involve the breast parenchyma, primary sarcoidosis of the breast is very rare. It occurs in less than 1% of cases. In a differential diagnosis it may potentially be considered a malignancy. CASE REPORT: We report a case in which breast sarcoidosis was the first clinical manifestation of systemic disease in a 54-year-old woman who presented with wide erythematous skin changes associated with palpable induration. Considering the fact that physical examination and the results of mammography, ultrasound and magnetic resonance imaging were inconclusive and unable to rule out malignancy, biopsy was performed. Pathohistological diagnosis showed a non-necrotizing granulomatous inflammation without elements of breast cancer. Sarcoidosis was confirmed with elevated level of angiotensin-converting enzyme in the sera and characteristic chest multislice computed tomography findings. The bronchoalveolar lavage was infiltrated with lymphocytes. CONCLUSION: Breast sarcoidosis has diverse and nonspecific imaging characteristics. Carcinoma must always be excluded by core needle biopsy. Achieving correct diagnosis is mandatory so that adequate corticosteroid therapy can be applied as early as possible. A multidisciplinary approach is of utmost importance in the diagnostic workup.