Detailed target-selectivity information and experiment-based efficacy prediction tools are primarily available for Streptococcus pyogenes Cas9 (SpCas9). One obstacle to develop such tools is the rarity of accurate data. Here, we report a method termed ‘Self-targeting sgRNA Library Screen’ (SLS) for assaying the activity of Cas9 nucleases in bacteria using random target/sgRNA libraries of self-targeting sgRNAs. Exploiting more than a million different sequences, we demonstrate the use of the method with the SpCas9-HF1 variant to analyse its activity and reveal motifs that influence its target-selectivity. We have also developed an algorithm for predicting the activity of SpCas9-HF1 with an accuracy matching those of existing tools. SLS is a facile alternative to the much more expensive and laborious approaches used currently and has the capability of delivering sufficient amount of data for most of the orthologs and variants of SpCas9.

Introduction and aim: The authors examined the pre-hospital delay of patients with ST-elevation myocardial infarction receiving percutaneous coronary revascularization. Method: In the Hungarian Myocardial Infarction Registry (HUMIR), between January 1, 2014 and March 31, 2016, 7146 patients were found who had all necessary time intervals available. In the database of the National Ambulance Service (OMSZ), 3288 patients were found who had the necessary time intervals. The following time intervals were investigated: the time from the beginning of the complaint to the rescue service notification (patient delay time = PDT), the time of the rescue service to arrive at the scene (R1), the on-site care time (R2) and the time from the scene until arriving to the centre (R3). The case of care at the centre, we investigated the time from the onset of symptoms until the balloon inflation (SBI). If the first hospital had no cardiac catheterization laboratory, we measured the transfer time to the cath centre. The methodological details related to the operation of the HUMIR had been described in our earlier communication. Rescue times (R1, R2, R3) were investigated on the basis of the paper-based records of the National Emergency Service. The patients were divided into two groups based on the fact that the first admission hospital is a centre with a heart catheter facility (C) or a non-invasive hospital (H). Results: 2621 patients (79.7%) were admitted to a hospital with cath lab (C) and 667 patients with secondary transport. Patients with primary transport to C were younger, but for other data, the two groups did not differ. The median of PDT for patients in group C was 114, and 121 minutes for patients in group H. There was no significant difference between R1 and R2 time between the two groups. R3 time in group C was longer than for H patients. In the case of secondary transport, the median time was 98 minutes until the centre. The median time from the beginning of the complaint to the balloon inflation (total ischemic time) was 260 minutes in group C and 356 minutes in group H. Conclusion: Based on the analysis of the total ischemic time and the pre-hospital delay, it is clear that the care of myocardial infarction patients can be further improved by reducing the patient’s decision time and increasing the proportion of primary transport. Rescue times (R1, R2, R3) met the expectations, however, further analysis of the M2 time should be considered. Orv Hetil. 2019; 160(1): 20–25.

Absztrakt: Bevezetés és célkitűzés: A szerzők 6878 acut myocardialis infarctus (AMI) miatt kezelt beteg sürgősségi ellátásának adatait elemzik a Nemzeti Szívinfarktus Regiszter és az Országos Mentőszolgálat adatainak felhasználásával. Módszer: A betegeknél 2017. 01. 01. és 2018. 12. 31. között katéteres érmegnyitás történt: 47,5%-uknál ST-elevációval járó szívinfarktus (STEMI), 3614 betegnél (52,5%) nem ST-elevációval járó szívinfarktus (NSTEMI) volt a kezelés indoka. Vizsgálták a panasz kezdete és a mentőszolgálat értesítése között eltelt időt (a betegek késlekedése), a mentés formáját (mentés, illetve szállítás). Elemezték a mentő helyszínre érkezésének (M1), a helyszíni ellátásnak (M2), a kórházba szállításnak (M3) az idejét, valamint a kórházi felvételtől az ér megnyitásáig eltelt időt. Az eredményeket megyék szerinti bontásban is közlik. A medián értékeket és a kvartiliseket (Q1, Q3) tüntették fel. Eredmények: A betegek késlekedése STEMI esetén 101, az NSTEMI-csoportban 687 perc volt. Mentés és azonnali mentőszállítás a STEMI-betegek 58,7%-ánál, az NSTEMI-betegek 43,7%-ánál történt. AMI esetén a mentőegység helyszínre érkezésének (M1) medián ideje 13 perc, a helyszíni ellátás (M2) 23 perc, a helyszínről a kórházba érkezésig eltelt idő (M3) 30 perc volt. STEMI esetén a felvételtől az ér megnyitásáig eltelt idő 37 perc volt. A STEMI-betegek 9,5%-ánál 2 órán belül, 49,1%-ánál 4 órán belül, 88,1%-ánál 12 órán belül került sor az infarktusért felelős ér megnyitására. Ebben a betegcsoportban a teljes ischaemiás idő medián értéke 243 perc volt. Minden vizsgált időperiódus esetén a megyék között jelentős különbségeket találtak. Következtetések: A szívinfarktusos betegek optimális időben történő ellátásának jelenleg a legnagyobb problémája a betegek késlekedése. A mentési idők tekintetében jelentős regionális különbségek vannak, az okok vizsgálatához további elemzések szükségesek. Orv Hetil. 2020; 161(12): 458–467.

One of the earliest population-based surveys regarding the incidence and treatment of myocardial infarction was initiated and coordinated by the World Health Organisation ~30 years ago. 1 Over the past two decades, many European and Far Eastern countries have launched registries covering the entire population, 2-5 and several comparative international studies have been performed. [6][7][8][9] Because of the varying methodologies of these registries, the comparative evaluation of data has only been possible with certain

Introduction: The significance of the total ischemic time (from the beginning of the complaint to the opening of the vessel) is an important factor for myocardial salvage. Aim: The aim of the study was to determine the prognostic significance of the TIT in patients with ST elevation myocardial infarction in Hungary. Method: From 1 January 2014 all patients with myocardial infarction were recorded by law in an on-line database of the Hungarian Myocardial Infarction Registry. Between 1 January 2014 and 31 March 2016, 27 157 patients with 28 408 myocardial infarction events were recorded. To investigate TIT, 7146 STEMI patients were selected who were treated with percutaneous coronary intervention (PCI) within 24 hours of the beginning of the complaint and all of its components were known. Results: Average follow-up was 740 ± 346 days. The median time of the TIT is 260 minutes, within which the earliest prehospital time was found (median 205 minutes). The TIT influenced survival: if this time was less than 400 minutes, the 30-day and the 1-year deaths were 7.5% and 12.2%, respectively. In longer TIT, higher mortality rate was found (9.2% versus 19.7%, respectively). Multivariate analysis was performed for short (<30 days), medium (30–364 days) and long-term (≥365 days) survival. Diabetes mellitus is a short-term prognostic factor, abnormal creatinine, and severe coronary status have affected short and medium survival. PCI was significant in terms of medium and long-term survival. Previous myocardial infarction and TIT influenced the long-term survival significantly. Conclusions: In Hungary, TIT is too long, and its dominant part falls within the prehospital period. The TIT is an independent prognostic factor, so reducing this time can improve the long-term prognosis of patients with ST-elevation myocardial infarction. Orv Hetil. 2018; 159(27): 1113–1120.