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.
Introduction: Traumatic brain injury is the leading cause of death in children and adults in developed countries. Severe traumatic brain injury is classified with Glasgow Coma Scale score 8 and less. About 50% of patients with severe traumatic brain injury developes at least one infection as a complication of primary condition during hospitalization in the Intensive Care Unit, resulting with fatal outcome in 28% of patients. Ventilator-associated pneumonia is the leading infection that affects patients with severe traumatic brain injury, with an incidence between 41% and 74%. Following are sepsis and urinary tract infections. The aim: To analyze the number of patients with nosocomial infection and isolated severe traumatic brain injury hospitalized in the Intensive Care Unit of the Clinical Hospital Centre Rijeka, Croatia, from 31 st January 2013 to 31 st December 2014. Patients and methods: A two-year retrospective study included 46 patients with isolated severe traumatic brain injury and nosocomial infection hospitalized in the Intensive Care Unit of the Clinical Hospital Centre Rijeka,Croatia, in the period from 31 st January 2013 to 31 st December 2014. All medical data was collected from the Division of Intensive Care Unit, Clinical Hospital Centre Rijeka, Croatia. Results: From 67 patients with isolated severe traumatic brain injury, 46 (68,65%) of them developed nosocomial infection. There was statistically significant more male patients than female (p < 0.05). The average age of infected patients was 57,8 years. The leading were the infections of the respiratory system. Gram-negative bacteria Proteus mirabilis and Pseudomonas aerugnonsa were the leading pathogens. The average duration of the infection was 5,77 day. Duration of mechanical ventilation accounted for the majority of the patients more than 10 days. The average duration of treatment for all 46 patients was 10,475 days, and for 16 (34,78%) of them, the treatment outcome was lethal. Conclusion: Nosocomial infections are becoming a major public health problem. The emphasis must be set on the prevention which includes maintaining the hygiene and the antiseptic rules among the medical personnel of the Intensive Care Unit. Already developed infections must be adequately treated so the negative treatment outcomes can be reduced.
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.
Transplantacija bubrega predstavlja najbolju metodu liječenja terminalne faze kronične bubrežne bolesti. Pripremu pacijenata za zahvat vodi tim u koji su uključeni nefrolog, kirurg – urolog, anesteziolog te liječnik obiteljske medicine, čija je međusobna suradnja nezaobilazni dio skrbi o pacijentu. U prijeoperacijskoj pripremi anesteziolog se susreće s brojnim komorbiditetima, kompleksnom anamnezom te širokim spektrom pacijentove medikamentozne terapije uz naglasak na kardiovaskularne bolesti koje predstavljaju najčešći uzrok smrtnosti u ovoj skupini pacijenata. Pojavnost hematoloških, metaboličkih, respiratornih i endokrinih bolesti česta je, te je iznimno važna optimalizacija pacijentova općeg stanja prije operacijskog zahvata. Transplantacija bubrega je zahvat koji se obično ne može planirati i prema njemu se treba ponašati kao prema hitnom zahvatu, čime se povećava rizik razvoja komplikacija. Za vrijeme samog zahvata koji se izvodi u općoj, balansiranoj anesteziji, uloga anesteziologa je poznavanje farmakokinetike i farmakodinamike lijekova te vođenje anestezije koja će omogućiti urednu funkciju perfuzije presatka, što se postiže održavanjem adekvatnih vrijednosti krvnog tlaka. Važno je pravovremeno započinjanje antibiotske i imunosupresivne terapije uz zadovoljavajuću analgeziju. U daljnjem poslijeoperacijskom tijeku nužno je rano prepoznavanje i prevencija mogućih komplikacija kako bi se spriječilo neželjeno odbacivanje bubrežnog presatka.
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