A phosphatidylinositol (4,5)-bisphosphate binding site within μ2-adaptin regulates clathrin-mediated endocytosis

The clathrin adaptor complex AP-2 serves to coordinate clathrin-coated pit assembly with the sorting of transmembrane cargo proteins at the plasmalemma. How precisely AP-2 assembly and cargo protein recognition at sites of endocytosis are regulated has remained unclear, but recent evidence implicates phosphoinositides, in particular phosphatidylinositol (4,5)-bisphosphate (PI[4,5]P2), in these processes. Here we have identified and functionally characterized a conserved binding site for PI(4,5)P2 within μ2-adaptin, the medium chain of the clathrin adaptor complex AP-2. Mutant μ2 lacking a cluster of conserved lysine residues fails to bind PI(4,5)P2 and to compete the recruitment of native clathrin/AP-2 to PI(4,5)P2-containing liposomes or to presynaptic membranes. Moreover, we show that expression of mutant μ2 inhibits receptor-mediated endocytosis in living cells. We suggest that PI(4,5)P2 binding to μ2-adaptin regulates clathrin-mediated endocytosis and thereby may contribute to structurally linking cargo recognition to coat formation

Variation in DNA methylation patterns of grapevine somaclones (Vitis vinifera L.)

<p>Abstract</p> <p>Background</p> <p>In traditional vine areas, the production should present a typicity that partly depends on the grapevine variety. Therefore, vine improvement is considered difficult because of the limited choice in the natural variability of the cultivars within the limits of their characteristics. A possibility to circumvent this problem is the use of somatic variability. <it>In vitro </it>somatic embryogenesis and organogenesis can lead to genotypic and phenotypic variations, described as somaclonal variation, that could be useful for the selection of improved grapevine genotypes.</p> <p>Results</p> <p>In order to study tissue culture-induced variation of grapevine, we have analysed 78 somaclones obtained from somatic embryos of two distinct cultivars using molecular marker techniques. SSRs were only useful to verify the conservation of the microsatellite genotype between the somaclones and the respective mother clones. AFLP polymorphism between mother clones and somaclones was 1.3–2.8 times higher to that found between clones. However, a majority of the somaclones (45/78) exhibited only few changes. Seven and five somaclones of 'Chardonnay 96' and 'Syrah 174', respectively, which covered at least all polymorphic loci found in AFLP analysis were used for MSAP study. All of the 120 polymorphic fragments were found only in the somaclones. The percentage of full methylation at CCGG recognition sites was slightly higher in somaclones due to more polymorphic bands generated after cleavage by <it>Eco</it>RI/<it>Hpa</it>II. Different digestion patterns revealed different methylation status, especially different levels of de-methylation, that are the consequence of the <it>in vitro </it>culture.</p> <p>Conclusion</p> <p>MSAP highlights DNA methylation variation in somaclones compared to mother clones and, therefore, is a powerful tool for genotypic characterisation of somatic embryo-derived grapevines. The detection of the same polymorphic bands in numerous somaclones of different cultivars suggests the possibility of hot spots of DNA methylation variation. SSR profiles of the 'Chardonnay' and 'Syrah' somaclones were the same as of the respective mother clones. The somaclones exhibited a higher AFLP variation than clones obtained via traditional clonal selection in the field. Therefore, somatic embryogenesis through <it>in vitro </it>culture technique could be useful for the selection of improved cultivars with subtle changes but conserving their main characteristics.</p

Wildbienen kennen lernen

Das vorliegende Unterrichtskonzept soll Schüler*innen der Sekundarstufe I Einsichten in die Lebensweise ausgewählter Wildbienen eröffnen. Die Lernenden befassen sich zunächst mit der bestäubungsbiologischen Bedeutung der Tiere. Dann lernen sie die Lebensformen von sechs exemplarischen Bienenarten bzw. –gattungen kennen. In einem weiteren Schritt befassen sie sich mit dem Lebenszyklus der Gehörnten Mauerbiene (Osmia cornuta). Zuletzt stellen sie eine eigene Wildbienen-Nisthilfe her

Az állam szerepe a rendszerváltásban

A központi tervgazdaságról a szabad piacgazdaságra való áttéréskor az állam szerepének a definiálása számos kelet-közép európai országban döntõnek bizonyult a reformfolyamat sikerességének szempontjából. Elsõ látásra az állam bilincseitõl való megszabadulás a reformországokban a gazdaság teljes állami befolyás alól való felszabadítását, és a független piaccal való helyettesítését jelentette. Szinte minden gazdasági aktor számára a sikeres nyugati ipari gazdaságok szimbóluma a szabad piacgazdaság volt. Csak fokozatosan látták be, hogy még a szabad piacgazdaság által szervezett gazdaságokban is szükség van egy elosztói szerepkört betöltõ gazdasági szereplõre. Az, hogy az állam a nemzetgazdaságban fontos allokációs funkcióval bír az erõforrások optimális elosztását illetõen, nehezen volt elmagyarázható azok számára, - gondoljunk példaként az NDK-ra - akik lépten-nyomon a szocialista tervgazdaság által hátrahagyott használhatatlan ipari létesítményekkel szembesültek, és a nyugati gazdaságokkal összevetve a lakosság alacsony életszínvonalát tapasztalták. A cikk az állam újragondolt szerepével foglalkozik a kiválasztott közép-kelet európai országokban. Az egyik fõ kérdés az, hogy mennyiben sikerült lefektetni a piacgazdaság konstitucionális elveit, mint ¢ magántulajdon, ¢ árképzés a versenypiacon, ¢ nyitott piacok, ¢ szerzõdési szabadság, ¢ szankciómechanizmusok a piacgazdasági felelõsségen keresztül, ¢ a gazdaságpolitika kiszámíthatósága. A másik kérdés, hogy mely szabályozási elveket soroltak a konstitucionális elvek mellé. Mindkét kérdés az államnak a piacgazdaságban betöltött szerepét érinti. Példaként említhetõ a fennálló állami szabályozás a magántulajdon és a gazdaság szereplõinek védelmében, vagy a versenykorlátozó ma- gatartás megakadályozása. Összefoglalva, az ilyen szabályozás az állami tevékenységen keresztül akkor szükséges, ha az egyén (gazdasági szereplõ) racionalitása a kollektív racionalitással (nemzetgazdasági szinten) nem egyezik. A cikk felsorol számos érvet amellett, hogy az államnak piacgazdasági versenyfeltételek között is a közösség érdekében kell tevékenykednie. A következõ szakaszban elõször pontosan meghatározzuk az állam szerepét, mielõtt a vizsgált országok bemutatásán keresztül az állam gyakorlatban betöltött szerepével foglalkoznánk

Anticipatory Inventory Management For Realizing Robust Production Processes In Engineer-To-Order Manufacturing: A Modeling Approach

At ever shorter intervals, manufacturing and processing companies of all industries are confronted with external or internal disruptions and crises that need to be managed. Consequently, a corporate focus on robust supply chains and processes is essential. At the same time, crises and their impact on supply chains cannot be predicted. To be able to act anticipatively, it is necessary to link product and production system design to take suitable measures to safeguard production at an early stage. In this context, a monetary conflict of objectives arises concerning when a company should position itself robustly and when it is sufficient to react flexibly to disruptions. The production planning and control (PPC) task inventory management is an essential lever for realizing robust order fulfilment processes. Inventory management aims to ensure that production and assembly within the company are supplied in the right quantities and without lateness. In particular, companies that operate according to the engineer-to-order strategy (ETO) face specific challenges in dimensioning stocks for materials or components - for example, due to the low level of standardization or lack of supplier diversity. This paper presents an approach for anticipatory inventory management using product portfolio characteristics. A new modeling approach for dimensioning safety stocks under the increasing influence of crises is also developed and integrated into the process

Incidence and challenges of helicopter emergency medical service (HEMS) rescue missions with helicopter hoist operations: analysis of 11,228 daytime and nighttime missions in Switzerland

Objective: We aimed to investigate the medical characteristics of helicopter hoist operations (HHO) in HEMS missions. Methods: We designed a retrospective study evaluating all HHO and other human external cargo (HEC) missions performed by Swiss Air-Rescue (Rega) between January 1, 2010, and December 31, 2019. Results: During the study period, 9,963 (88.7 %) HEMS missions with HHO and HEC were conducted during the day, and 1,265 (11.3 %) at night. Of the victims with time-critical injuries (NACA ≥ 4), 21.1 % (n = 400) reached the hospital within 60 min during the day, and 9.1 % (n = 18) at night. Nighttime missions, a trauma diagnosis, intubation on-site, and NACA Score ≥ 4 were independently and highly significantly associated with longer mission times (p < 0.001). The greatest proportion of patients who needed hoist or HEC operations in the course of the HEMS mission during the daytime sustained moderate injuries (NACA 3, n = 3,731, 37.5 %) while practicing recreational activities (n = 5,492, 55.1 %). In daytime HHO missions, the most common medical interventions performed were insertion of a peripheral intravenous access (n = 3,857, 38.7 %) and administration of analgesia (n = 3,121, 31.3 %). Conclusions: Nearly 20 % of patients who needed to be evacuated by a hoist were severely injured, and complex and lifesaving medical interventions were necessary before the HHO procedure. Therefore, only adequately trained and experienced medical crew members should accompany HHO missions

Oral transmucosal fentanyl citrate analgesia in prehospital trauma care: an observational cohort study.

BACKGROUND Pain is one of the major prehospital symptoms in trauma patients and requires prompt management. Recent studies have reported insufficient analgesia after prehospital treatment in up to 43% of trauma patients, leaving significant room for improvement. Good evidence exists for prehospital use of oral transmucosal fentanyl citrate (OTFC) in the military setting. We hypothesized that the use of OTFC for trauma patients in remote and challenging environment is feasible, efficient, safe, and might be an alternative to nasal and intravenous applications. METHODS This observational cohort study examined 177 patients who were treated with oral transmucosal fentanyl citrate by EMS providers in three ski and bike resorts in Switzerland. All EMS providers had previously been trained in administration of the drug and handling of potential adverse events. RESULTS OTFC caused a statistically significant and clinically relevant decrease in the level of pain by a median of 3 (IQR 2 to 4) in NRS units (P < 0.0001). Multiple linear regression analysis showed a significant absolute reduction in pain, with no differences in all age groups and between genders. No major adverse events were observed. CONCLUSIONS Prehospital administration of OTFC is safe, easy, and efficient for extrication and transport across all age groups, gender, and types of injuries in alpine environments. Side effects were few and mild. This could provide a valuable alternative in trauma patients with severe pain, without the delay of inserting an intravenous line, especially in remote areas, where fast action and easy administration are important

Airway management in a Helicopter Emergency Medical Service (HEMS): a retrospective observational study of 365 out-of-hospital intubations.

BACKGROUND Airway management is a key skill in any helicopter emergency medical service (HEMS). Intubation is successful less often than in the hospital, and alternative forms of airway management are more often needed. METHODS Retrospective observational cohort study in an anaesthesiologist-staffed HEMS in Switzerland. Patient charts were analysed for all calls to the scene (n = 9,035) taking place between June 2016 and May 2017 (12 months). The primary outcome parameter was intubation success rate. Secondary parameters included the number of alternative techniques that eventually secured the airway, and comparison of patients with and without difficulties in airway management. RESULTS A total of 365 patients receiving invasive ventilatory support were identified. Difficulties in airway management occurred in 26 patients (7.1%). Severe traumatic brain injury was the most common indication for out-of-hospital Intubation (n = 130, 36%). Airway management was performed by 129 different Rega physicians and 47 different Rega paramedics. Paramedics were involved in out-of-hospital airway manoeuvres significantly more often than physicians: median 7 (IQR 4 to 9) versus 2 (IQR 1 to 4), p < 0.001. CONCLUSION Despite high overall success rates for endotracheal intubation in the physician-staffed service, individual physicians get only limited real-life experience with advanced airway management in the field. This highlights the importance of solid basic competence in a discipline such as anaesthesiology