CRISPR-Cas in Escherichia coli: regulation by H-NS, LeuO and temperature

CRISPR-Cas adaptive immune systems are present in many bacteria and archaea and provide protection against invading DNA such as phages and plasmids. These systems are very versatile and complex in their gene composition and genomic architecture. CRISPR-Cas systems are classified into 2 classes, 6 types and 33 subtypes although this number is not definitive and the research is ongoing. All CRISPR-Cas systems have been thoroughly investigated in order to better understand the mechanism of CRISPR immunity enabling its use as a tool in genome editing and other biotechnological applications. However, regulation of the CRISPR-Cas system is also very complex and still not fully understood; it must provide optimal protection without introducing harmful consequences to the host. In this review we give an overview on the regulation of the CRISPR-Cas system Class 1 Type I-E in Escherichia coli with the emphasis on the role of temperature in regulation of the CRISPR-Cas activity and the interplay of the key regulators H-NS and StpA repressors and LeuO antirepressor in regulation of cas gene expression and HtpG chaperone in maintaining functional levels of Cas3.</p

Teaching nursing art using literature, music and painting

Od sredine 20. stoljeća kroz izvorne sestrinske teorije pokušava se odrediti autentičnost sestrinstva kao discipline. Ono se uglavnom utvrđuje u odmaku od klasične biomedicinske paradigme koja je vezana za medicinu i liječničku profesiju. Već je Florence Nightingale taj odmak smatrala nužnim, a utvrdila ga je u holističkom pristupu čovjeku koji je odredila pomoću koncepta umijeća. Mnoge su druge teoretičarke, počevši od Peplau 1952. godine, ono autentično u sestrinstvu odredile u spoznajama koje su između empirijskog i neempirijskog, odnosno ono što će Cooper nazvati posebnim znanjima u sestrinstvu. Budući da se radi o teorijskim modelima i konceptima koji se razvijaju uglavnom na američkom tlu tijekom proteklih desetljeća, njihov je utjecaj još na razini razvijanja jedinstvenih hipoteza i sporadičnih istraživanja. Ipak, fenomeni koji obuhvaćaju te vrste spoznaja proizlaze iz neposrednog iskustva i doživljaja čovjeka u bolesti ili pri povratku zdravlja, stoga su u domeni subjektivnih spoznaja. S jedne su strane ta iskustva teška za konceptualiziranje, a time i istraživanje, a s druge su iznimno izazovna za poučavanje. Prijedlog autora je da se, utemeljeno na dostupnoj literaturi i pozitivnim primjerima iz prakse, kao metoda poučavanja za osjećaj suosjećanja i razumijevanja iskustva uvedu sadržaji umjetnosti kako to predlaže Watson, odnosno da se izlaganjem učenika i studenata umjetničkim sadržajima potakne njihova senzibilizacija za emotivna reagiranja sa svim pozitivnim posljedicama istih.Since the middle of the 20th century, through original nursing theories, attempts have been made to determine the authenticity of nursing as a discipline. It is mainly established in a departure from the classic biomedical paradigm, which is related to medicine and the medical profession. Florence Nightingale already considered this step necessary and established it in a holistic approach to man, which she defined through the concept of skill. Many other theoreticians, starting with Peplau in 1952, determined that authenticity in nursing is in the knowledge between empirical and non-empirical, that is, what Cooper will call special knowledge in nursing. Given that these are theoretical models and concepts that have been developed mainly on American soil in the past decades, their influence is still at the level of developing unique assumptions and hypotheses and sporadic research. Nevertheless, the phenomena that comprise these types of knowledge arise from the immediate experience of a person in illness or when recovering from health and are, therefore, in the domain of subjective knowledge. On the one hand, these experiences are difficult to conceptualize and research, and on the other hand, they are extremely challenging to teach. The author proposes that, based on the available literature and positive examples from practice, as a teaching method for the feeling of compassion and understanding of experience, the contents of art should be introduced as suggested by Watson, for example, by exposing pupils and students to artistic content, their sensitization to emotional reactions with all positive consequences thereof

Obostrani pneumotoraks kao komplikacija perkutane traheotomije: prikaz slučaja

Percutaneous dilatational tracheostomy is a common surgical procedure that is becoming the method of choice in critically ill patients whenever prolonged airway secure and/ or ventilation support is needed. Although adverse events are relatively uncommon, serious life threatening complications can arise from this bedside procedure. We report a case of a 70-year-old female who developed extensive subcutaneous emphysema and bilateral pneumothorax immediately after a percutaneous dilatational tracheostomy procedure. Different mechanisms, such as damage to posterior or anterior tracheal wall, false passage or paratracheal placement or dislocation of the cannula are considered to be responsible for the development of pneumothorax and subcutaneous emphysema. Although bronchoscopic control after the tracheostomy procedure did not reveal any tracheal injury, we believe that subcutaneous emphysema and bilateral pneumothorax are most likely caused by procedure induced injuries of the trachea in addition to the applied high airway pressure induced by excessive or inappropriate ventilation. In our case report, we would like to emphasize that continuous bronchoscopic guidance during percutaneous tracheostomy is invaluable in decreasing the incidence of its overall complications, especially during enhancing the team experience.Perkutana dilatacijska traheotomija je učestali kirurški zahvat koji postaje metodom osiguravanja dišnoga puta u kritičnih bolesnika i bolesnika na dugotrajnoj mehaničkoj ventilaciji. Iako su popratni učinci relativno rijetki, metoda je vezana uz mogućnost nastanka za život opasnih komplikacija. Prikazujemo slučaj 70-godišnje žene kod koje je došlo do razvoja opsežnog potkožnog emfizema i obostranog pneumotoraksa neposredno nakon izvođenja perkutane traheotomije. Različiti mehanizmi kao što su oštećenje stražnjeg ili prednjeg zida dušnika, lažni prolaz ili paratrahealno postavljanje ili dislokacija kanile mogu se smatrati odgovornim za nastanak pneumotoraksa i potkožnog emfizema. U našem prikazu slučaja, iako nakon perkutane traheotomije bronhoskopskim pregledom nije nađeno oštećenje dušnika, smatramo da je uzrok nastanka potkožnoga emfizema i obostranog pneumotoraksa najvjerojatnije procedurom nastala ozljeda dušnika i primjena visokog tlaka u dišnim putovima izazvanog prejakom ili neodgovarajućom ventilacijom. Ovim prikazom slučaja željeli bismo naglasiti važnost bronhoskopske vizualizacije tijekom izvođenja perkutane traheotomije u prevenciji nastanka mogućih komplikacija, osobito u razdoblju usvajanja ove kliničke vještine

POSTOPERATIVE ANALGESIA FOLLOWING RECONSTRUCTIVE KNEE URGERY: COMPARISON OF DIFFERENT PERIPHERAL NERVE BLOCK COMBINATIONS

Cilj: Prikazati jačinu rane poslijeoperacijske boli u bolesnika podvrgnutih rekonstrukcijskom zahvatu prednje ukrižene sveze koljena u neuroaksijalnoj anesteziji primjenom subarahnoidnog bloka uz primjenu kombinacije analgetskog bloka femoralnog živca u aduktornom kanalu s poplitealnim blokom ishijadikusa ili kombinacije bloka femoralnog živca u aduktornom kanalu s transglutealnim blokom ishijadikusa. Metode: U ovoj prospektivnoj studiji bolesnici su randomizirani u dvije skupine ovisno o vrsti primijenjenih perifernih živčanih blokova. Skupinu I (n=20) činili su bolesnici analgezirani kombinacijom bloka femoralnog živca u aduktornom kanalu i poplitealnog bloka ishijadičnog živca (AP), a skupinu II (n=20) bolesnici analgezirani kombinacijom bloka femoralnog živca u aduktornom kanalu i transglutealnog bloka ishijadičnog živca (AG). U svih bolesnika blokovi su izvedeni pod kontrolom ultrazvuka primjenom 0,25 % levobupivakaina uz dodatak 4 mg deksametazona za svaki pojedinačni blok. Nakon primjene perifernih živčanih blokova u svih je bolesnika primijenjena subarahnoidna anestezija. Mjerili smo vrijeme potrebno za izvođenje blokova, vrijeme do nastanka pojedinog bloka promjenom reakcije na hladno, razinu akutne poslijeoperacijske boli na 10-centimetarskoj vizualno analognoj ljestvici (VAS) neposredno nakon zahvata te 4, 8 i 24 sata nakon zahvata). Bilježili smo nastanak neželjenih nuspojava, ukupnu potrošnju dodatnih analgetika i zadovoljstvo bolesnika nakon provedenih postupaka numeričkom ljestvicom 1-10. Rezultati: Ispitanici su bili u dobi 18-30 godina, a 70 % su bili muškarci. Između skupina AP i AG nisu zabilježene statistički značajne razlike za jačinu boli ni u jednom promatranom vremenskom intervalu (AP VAS 2,70 vs AG VAS 2,35, 24 sata poslijeoperacijski). Vrijeme potrebno za izvođenje blokova bilo je značajno kraće u AP skupini (AP 7,45 min ± 1,05 vs AG 10,60 min ± 2,11). Utvrđene su značajne razlike u prosječnom vremenu potrebnom za nastup bloka ishijadikusa ovisno o tome je li blok izveden poplitealnim (P) ili transglutealnim pristupom (G) (P 4,45 min vs G 7,80 min). U skupini AG šest bolesnika se izjasnilo da je postupak transglutealnog pristupa bio izrazito bolan. U skupini AP nisu zabilježene komplikacije, dok je u skupini AG jedan bolesnik imao mučninu, a jedan je otežano mokrio. Razlike u učestalosti primjene dodatnih analgetika između promatranih skupina nisu bile statistički značajne. Ukupno zadovoljstvo bolesnika nakon provedenih postupaka bilo je značajno veće u AP skupini (AP 9,35 ± 0,67 vs AG 8,65 ± 0,93). Zaključak: Obje kombinacije perifernih živčanih blokova osigurale su zadovoljavajuću razinu poslijeoperacijske analgezije. Međutim, u AG skupini ukupno zadovoljstvo bolesnika nakon provedenih postupaka bilo je niže zbog duljeg trajanja izvođenja i bolnosti. Kombinacija aduktornog i poplitealnog bloka osigurala je dobru analgeziju bez značajnih nuspojava i bez poslijeoperacijske upotrebe opioida i ova kombinacija može koristiti kao dio multimodalne analgezije u rekonstrukcijskim zahvatima koljena.The aim was to compare the quality of postoperative analgesia and incidence of adverse effects using two different peripheral nerve block combinations in patients undergoing reconstructive knee surgery under spinal anesthesia. This prospective study included 40 patients undergoing elective reconstructive knee surgery under spinal anesthesia who were randomly divided into two groups of 20 patients each. Patients in AP group received ultrasound guided adductor canal and popliteal block and patients in AG group received ultrasound guided adductor canal and transgluteal sciatic nerve block. Blocks were performed in all patients using 0.25% levobupivacaine and 4 mg dexamethasone for each block (20 mL total LA volume for transgluteal and popliteal block, 10 mL for adductor canal block). We measured and observed the time needed to perform the blocks, time to each block onset (changed reaction to cold using a cotton swab dipped in medicinal alcohol), postoperative pain level according to 10-cm visual analog scale (VAS; immediately after surgery and 4, 8 and 24 hours postoperatively), adverse effects, total analgesic consumption, and patient overall satisfaction on 1-10 numerical scale. Patients included in the study were 18-30 years old, 70% of them were male. There was no statistically signifi cant difference in VAS scores between the groups during study protocol (mean AP VAS 2.70 vs. AG VAS 2.35 24 hours postoperatively). The time necessary to perform the blocks was signifi cantly shorter in AP group compared to AG group (mean 7.45±1.05 min vs. 10.60±2.11 min). Time to sciatic nerve block onset was signifi cantly shorter in AP group (mean AP 4.45 min vs. AG 7.80 min). Six patients in AG group reported transgluteal approach to be very painful. We recorded no adverse effects in AP group. In AP group, one patient reported nausea and urinary retention each. There was no signifi cant difference in requiring additional analgesia between the groups. Patient overall satisfaction was signifi cantly higher in AP group (mean 9.35 vs. 8.65). In conclusion, both peripheral nerve block combinations provided adequate level of postoperative analgesia but adductor canal block and transgluteal sciatic nerve block combination required longer execution time, patients reported transgluteal application to be painful, and their verall satisfaction was lower. The combination of adductor canal block and popliteal block provided adequate analgesia with no adverse effects and no postoperative opioid consumption, and this combination can be used as part of multimodal analgesia for reconstructive knee surgery

Preliminarni rezultati istraživanja entomofaune sjemena poljskog (Fraxinus angustifolia Vahl) i pensilvanskog jasena (Fraxinus pennsylvanica Marshall) u Hrvatskoj

Native narrow-leaved ash (Fraxinus angustifolia Vahl) and introduced green ash (F. pennsylvanica Marshall) are two most common ash (Fraxinus spp.) species in Croatian lowland forests, taking a significant place in these ecosystems. In the recent past it has been observed that narrow-leaved ash seed yields have reduced, emphasising the importance of good health status of those collected, as they are used for seedling production and consecutive regeneration of forest stands or afforestation. Given that insects could be an important factor affecting ash seed health status, the aim of this research was to gain insight into the species being present and their frequency in the ash seeds. In total 2.500 narrow-leaved ash and 1.000 green ash seeds from seven different locations were screened for insect presence. Analysed seeds were categorized as undamaged, insect damaged or empty, and observed insect specimens were identified by morphological and molecular genetic analysis. Green ash seeds revealed higher insect infestation in comparison with native ash species, with weevil Lygniodes bischoffii being the most numerous pest found. Narrow-leaved ash seeds were mostly occupied by the native weevil species, L. enucleator. Besides these pests, larval, pupal, and adult stages of parasitoid wasps (Hymenoptera) and larval stage of the seed eating moth Pseudargyrotoza conwagana (Lepidoptera) were found in both ash species. However, given that pests detected in this study prefer urban areas or were present in a very small number of seeds, where they were often followed by parasitoids, it can be concluded that insects currently do not have a significant negative impact on the reproduction of narrow-leaved ash and green ash in forest stands.Autohtoni poljski jasen (Fraxinus angustifolia Vahl) i alohtoni pensilvanski jasen (F. pennsylvanica Marshall) su dvije najčešće vrste roda Fraxinus prisutne u nizinskim šumama Hrvatske, zbog čega zauzimaju važno mjesto u ovim šumskim ekosustavima. Posljednjih godina je zapaženo smanjenje uroda sjemena poljskog jasena, naglašavajući važnost dobrog zdravstvenog stanja sakupljenih količina, jer se sjeme koristi za proizvodnju sadnica koje se dalje upotrebljavaju za pomlađivanje šumskih sastojina ili pošumljavanje. Kako kukci mogu imati velik utjecaj na zdravstveno stanje sjemena, cilj ovog istraživanja je bio dobiti uvid u prisutne vrste i njihovu učestalost u sjemenu jasena. Prisutnost kukaca je analizirana u ukupno 2.500 sjemenki poljskoga i 1.000 sjemenki pensilvanskog jasena. Promatrano sjeme je kategorizirano kao neoštećeno, oštećeno i prazno (šturo), a nađene jedinke kukaca su identificirane na temelju morfoloških karakteristika i korištenjem molekularno genetičke analize. Sjeme pensilvanskog jasena pokazalo je veću oštećenost kukcima u usporedbi sa sjemenom poljskoga jasena, pri čemu je pipa Lygniodes bischoffii bila najčešći utvrđeni štetnik. Oštećeno sjeme poljskog jasena je najčešće bilo nastanjeno autohtonom vrstom pipe, L. enucleator. Osim navedenih štetnika, u sjemenu obje vrste jasena su nađene ličinke, kukuljice i imaga parazitoidnih osa (Hymenoptera) te ličinke štetnika Pseudargyrotoza conwagana (Lepidoptera). S obzirom da štetnici utvrđeni u istraživanju preferiraju urbana područja ili su nađeni na vrlo malom broju sjemena, često zajedno sa parazitoidima, može se zaključiti kako kukci trenutno nemaju značajan negativan utjecaj na zdravstveno stanje sjemena poljskog i pensilvanskog jasena u šumskim sastojinama

NONINVASIVE VENTILATION IN THE POSTOPERATIVE PERIOD

Akutno zatajenje disanja relativno je česta poslijeoperacijska komplikacija, a najčešće se javlja u bolesnika s prijeoperacijski poremećenom plućnom funkcijom i/ili nakon velikih torako-abdominalnih operacija. Iako se endotrahealna intubacija i mehanička ventilacija još uvijek smatraju osnovom u liječenju bolesnika s akutnim zatajenjem disanja, literaturni podatci ukazuju da je mortalitet tih bolesnika usko povezan s komplikacijama vezanim uz intubaciju i mehaničku ventilaciju. Zbog toga je cilj spriječiti komplikacije i ako je moguće izbjeći endotrahealnu intubaciju. Neinvazivna ventilacija (NIV) je oblik mehaničke potpore koja ne zahtijeva postavljanje endotrahealnog tubusa te se danas sve češće koristi u proilaktičke i terapijske svrhe kod kirurških bolesnika s akutnim zatajenjem disanja. U ovom članku dajemo kratak pregled indikacija i ograničenja neinvazivne ventilacije u perioperacijskom razdoblju.Acute respiratory failure is relatively common postoperative complication, occurs frequently in patients with preexisting compromised respiratory function and/or undergoing major thoracoabdominal procedures. Although endotracheal intubation and mechanical ventilation is still considered the cornerstone for the treatment of patients with acute respiratory failure, literature data have shown that mortality of these patients is closely associated to complications of intubation and mechanical ventilation. Therefore, the goal is to prevent complications and if possible to avoid endotracheal intubation. Noninvasive ventilation (NIV) is a form of mechanical support that does not require endotracheal intubation and is nowadays often used in prophylactic and therapeutic purposes in surgical patients with acute respiratory failure. In this article we give a brief overview of the indications and limitations of non-invasive ventilation in perioperative period

Stanična imunost kunića nakon višekratne izloženosti sevofluranu i dušikovu oksidulu

The aim of this study was to investigate the effect of repeated/prolonged exposure to sevoflurane and nitrous oxide on the immune response in rabbits. Venous blood samples were obtained from twenty New Zealand rabbits, randomly allocated to four groups of five rabbits each. Group A rabbits served as controls, breathing 100% oxygen, group B rabbits inhaled a mixture of oxygen and nitrous oxide, group C inhaled a combination of oxygen and sevoflurane, and group D inhaled a combination of oxygen, nitrous oxide and sevoflurane. Flow cytometry was used to determine the proportions of CD45+, CD4+, CD8+ and CD21+ leukocytes subpopulations. Peripheral lymph nodes, lungs and spleen were sampled and subjected to histologic examination. The greatest changes in the proportion of CD21+, CD4+, CD8+ and CD45+ cells were recorded in groups C and D on days 7, 14 and 21. Comparison of the CD21+, CD4+, CD8+ and CD45+ cell counts on day 0 and day 21 showed a 30% decrease. This study demonstrated that sevoflurane alone and in combination with nitrous oxide exerts a strong immunosuppressive effect, modifying the physiological values of the hematologic parameters, and inducing histopathological alterations in peripheral lymph node, lung and spleen tissues.Kirurški zahvati i anestezija mogu imati znatan učinak na imunosni sustav. S obzirom na to da određeni kirurški postupci zahtijevaju opetovanu anesteziju, u ovom smo istraživanju pratili učinak višekratnog izlaganja sevoflurana i dušikova oksida na imunosni sustav kunića. Dvadeset novozelandskih kunića podijeljeno je slučajnim izborom u četiri skupine. U svakoj je skupini bilo 5 životinja. Istraživanje je provedeno u trajanju od 20 minuta dnevno tijekom 21 dana. Kunići skupine A, udišući 100 %-tni kisik, služili su kao kontrolna skupina. U skupini B kunići su udisali mješavinu kisika i dušikova oksida, u skupini C primijenjena je kombinacija kisika i sevoflurana, dok je u skupini D korištena kombinacija kisika, dušikova oksida i sevoflurana. Uzorci krvi prikupljeni su 3., 7., 14. i 21. dan za kompletnu krvnu sliku i za određivanje subpopulacije leukocita na protočnom citometru (CD21+, CD4+, CD8+, CD45+). Nakon završetka istraživanja sve su životinje žrtvovane. Uzorci perifernog limfnog čvora, pluća i slezene uzeti su za histološku analizu. Najočitije promjene uočene su u promjeni udjela CD21+, CD4+, CD8+, CD45+ stanica u skupinama C i D 7., 14. i 21. dan istraživanja. Ako se usporedi vrijednost udjela CD21+, CD4+, CD8+, CD45+ stanica nulti dan i 21. dan istraživanja, uočava se pad udjela za približno 30 %. U ovom je istraživanju dokazano da sevofluran, te sevofluran u kombinaciji s dušikovim oksidulom, ima izrazit imunosupresijski učinak te utječe na promjenu fizioloških vrijednosti hematoloških pokazatelja i pojavu histopatoloških promjena na tkivima perifernog limfnog čvora, plućiju i slezene

Harmonised projections of future forest resources in Europe

Data PaperAbstract • Key message A dataset of forest resource projections in 23 European countries to 2040 has been prepared for fores trelated policy analysis and decision-making. Due to applying harmonised definitions, while maintaining country-specific forestry practices, the projections should be usable from national to international levels. The dataset can be accessed at https://doi.org/10.5061/dryad.4t880qh. The associated metadata are available at https://metadata-afs.nancy.inra.fr/ geonetwork/srv/eng/catalog.search#/metadata/8f93e0d6-b524-43bd-bdb8-621ad5ae6fa9info:eu-repo/semantics/publishedVersio

Harmonised statistics and maps of forest biomass and increment in Europe.

peer reviewedForest biomass is an essential resource in relation to the green transition and its assessment is key for the sustainable management of forest resources. Here, we present a forest biomass dataset for Europe based on the best available inventory and satellite data, with a higher level of harmonisation and spatial resolution than other existing data. This database provides statistics and maps of the forest area, biomass stock and their share available for wood supply in the year 2020, and statistics on gross and net volume increment in 2010-2020, for 38 European countries. The statistics of most countries are available at a sub-national scale and are derived from National Forest Inventory data, harmonised using common reference definitions and estimation methodology, and updated to a common year using a modelling approach. For those counties without harmonised statistics, data were derived from the State of Europe's Forest 2020 Report at the national scale. The maps are coherent with the statistics and depict the spatial distribution of the forest variables at 100 m resolution