The cryo-electron microscopy structure of huntingtin.

Huntingtin (HTT) is a large (348 kDa) protein that is essential for embryonic development and is involved in diverse cellular activities such as vesicular transport, endocytosis, autophagy and the regulation of transcription. Although an integrative understanding of the biological functions of HTT is lacking, the large number of identified HTT interactors suggests that it serves as a protein-protein interaction hub. Furthermore, Huntington's disease is caused by a mutation in the HTT gene, resulting in a pathogenic expansion of a polyglutamine repeat at the amino terminus of HTT. However, only limited structural information regarding HTT is currently available. Here we use cryo-electron microscopy to determine the structure of full-length human HTT in a complex with HTT-associated protein 40 (HAP40; encoded by three F8A genes in humans) to an overall resolution of 4 Å. HTT is largely α-helical and consists of three major domains. The amino- and carboxy-terminal domains contain multiple HEAT (huntingtin, elongation factor 3, protein phosphatase 2A and lipid kinase TOR) repeats arranged in a solenoid fashion. These domains are connected by a smaller bridge domain containing different types of tandem repeats. HAP40 is also largely α-helical and has a tetratricopeptide repeat-like organization. HAP40 binds in a cleft and contacts the three HTT domains by hydrophobic and electrostatic interactions, thereby stabilizing the conformation of HTT. These data rationalize previous biochemical results and pave the way for improved understanding of the diverse cellular functions of HTT.

Drugging an undruggable pocket on KRAS.

The 3 human RAS genes, KRAS, NRAS, and HRAS, encode 4 different RAS proteins which belong to the protein family of small GTPases that function as binary molecular switches involved in cell signaling. Activating mutations in RAS are among the most common oncogenic drivers in human cancers, with KRAS being the most frequently mutated oncogene. Although KRAS is an excellent drug discovery target for many cancers, and despite decades of research, no therapeutic agent directly targeting RAS has been clinically approved. Using structure-based drug design, we have discovered BI-2852 (1), a KRAS inhibitor that binds with nanomolar affinity to a pocket, thus far perceived to be "undruggable," between switch I and II on RAS; 1 is mechanistically distinct from covalent KRASG12C inhibitors because it binds to a different pocket present in both the active and inactive forms of KRAS. In doing so, it blocks all GEF, GAP, and effector interactions with KRAS, leading to inhibition of downstream signaling and an antiproliferative effect in the low micromolar range in KRAS mutant cells. These findings clearly demonstrate that this so-called switch I/II pocket is indeed druggable and provide the scientific community with a chemical probe that simultaneously targets the active and inactive forms of KRAS.

[Pneumology in the Model Degree Program of the Medical Faculty Ostwestfalen Lippe, Germany]. / Das Fach Pneumologie im Modellstudiengang der Medizinischen Fakultät Ostwestfalen-Lippe (OWL).

The specialist field of "pneumology" is still underrepresented in university clinics in Germany, but this is not the case at the newly founded medical faculty Ostwestfalen-Lippe (OWL) in Bielefeld. This is linked to representing pneumology and internal intensive care medicine in patient care, teaching and research across the board and the opportunity to actively help shape the development of the human medicine faculty in an exciting environment.The early anchoring of the subject "Pneumology" in the model degree program of medical school in OWL (begin winter semester 2021/22) contributes to further visibility and a university medical orientation. In this overview various issues of Pneumology in the Model Degree Program are explored by basic scientists, clinical teachers, members of the medical faculty and a student.In today's Evangelisches Klinikum Bethel (EvKB), pulmonary medicine has a long tradition. The hospital's first lung and infection center was opened in 1927. The EvKB's department for internal medicine, pneumology and intensive care medicine, which has been independent since 2009, is becoming a university clinic for pneumology within the medical faculty OWL. Relevant translational and interdisciplinary research can be intensified.There are 30 "Pneumology" teaching units in the model degree program, which are divided into two study sections using different formats, such as lectures, seminars, hands on courses and skills lab. It is represented in particular in the module complex "Circulation and Respiration". The content of the first phase of teaching was carried out by a module commission, with members representing the subjects involved in the module.Knowledge of the basics from, for example, physiology, pathophysiology, anatomy and pathology are taught to the students in the run-up to the pneumology course. Using the example of physiology, the presentation of the learning content of a basic subject is elaborated in this article.Half of all teaching units on pneumology of the entire course took place in the 2nd semester (in March and April 2022), so that students experienced the clinical relevance of the content at an early stage. There was a particular focus on obstructive airway and restrictive lung diseases. After imparting the basic knowledge of the physical examination of the lungs in the Skills Lab, the most important pathological findings in the above-mentioned diseases on inspection, palpation, auscultation and percussion are demonstrated and practised in patients as part of bedside teaching under supervision.Communication training is also longitudinally integrated into the modular teaching, with a total of more than 200 teaching hours and is performed interdisciplinary. In the "Circulation and Breathing" module eight hours are devoted to this with simulated patients, the anamnesis and therapy advice on classic cardiopulmonary diseases. For the students, integrating the teaching of basic theory and its clinical application for each organ systems represents a challenge in the model degree program, the advantages outweigh from today's perspective.

Hemolysis rates in blood samples: differences between blood collected by clinicians and nurses and the effect of phlebotomy training.

BACKGROUND: Hemolytic samples are one of the most challenging preanalytical issues in laboratory medicine. Even causes leading to hemolytic specimen are various, including phlebotomy practices. Respective educational interventions as well as the reduction of the number of people involved in blood collections are claimed to influence the sample quality for the better. In our hospital 70 junior doctors were in charge of routine phlebotomy until 2012, when this task was shifted to 874 nurses, including a preceding training in phlebotomy and preanalytics. Our aim was to evaluate the impact of this training effect and the increase of people involved on sample quality. METHODS: The hemolysis index (HI) of 43,875 samples was measured before (n=21,512) and after (n=22,363) the switch of blood collection responsibilities. Differences in overall hemolysis rates and the amount of plasma samples with a concentration of free hemoglobin (fHb) above 0.5 g/L and 1 g/L were calculated. RESULTS: Overall HI as well as the percentage of samples with an fHb concentration >0.5 g/L decreased after the responsibility for phlebotomy changed. The rate of samples with an fHb concentration >1 g/L remained unchanged. CONCLUSIONS: Hemolysis rates were reduced upon passing phlebotomy tasks from untrained physicians on to a trained nursing staff. We therefore conclude that the number of people performing phlebotomy seems to play a minor role, compared to the effect of a standardized training. However, whether a reduction in the number of people involved in blood collection could lead to further improvement of sample quality, remains to be investigated in future studies.

Non-Local Muscle Fatigue Impairs Mean Propulsive Velocity During Strength Loading in Strength-Trained Men.

Purpose: This study examined the acute influence of a bench press (BP) loading on the explosive squat (SQ) performance and vice versa. Methods: Nineteen strength-trained men completed 2 experimental sessions consisting of either a SQ+BP loading or a BP+SQ loading with 3 × 5 + 3 × 3 repetitions at 80% of the 1-repetition maximum in a randomized order. SQ and BP mean propulsive velocity (MPV) were assessed during both loadings, at baseline (T0) as well as immediately after the first (T1) and second strength loading (T2). Results: Both BP and SQ MPV decreased between T0 and T1 in SQ+BP (-6.13 ± 6.13%, p = .014, g = 0.485 and -9.11 ± 7.23%, p < .001, g = 0.905, respectively) and BP+SQ (-15.15 ± 7.69%, p < .001, g = 1.316 and -7.18 ± 6.16%, p < .001, g = 0.724, respectively). Mean BP MPV was lower in set 2 to set 6 in SQ+BP when compared to BP+SQ (-7.90% - 9.88%, all p < .05, g = 0.523-0.808). Mean SQ MPV was lower in set 1 and set 4 in BP+SQ when compared to SQ+BP (-4.94% - 5.22%, all p < .001, g = 0.329-0.362). Conclusions: These results demonstrate that the presence of non-local muscle fatigue affects the movement velocity. Therefore, if a training program aims to perform strength training exercises with maximal movement velocity, it is essential to carefully evaluate whether upper and lower body exercises should be carried out within close proximity.

Influence of menstrual cycle and oral contraceptive phases on strength performance, neuromuscular fatigue, and perceived exertion.

The primary aim of the study was to assess differences in strength performance, neuromuscular fatigue, and perceived exertion across phases of the menstrual cycle [MC; early follicular (eFP), late follicular (lFP), and mid-luteal phase (mLP)] and oral contraceptives [OCs; active pill phase (aPP) and nonactive pill phase (nPP)]. The secondary aim was to analyze the influence of fluctuating serum 17ß-estradiol and progesterone concentrations on these parameters in naturally menstruating women. Thirty-four women (21 with a natural MC and 13 using OCs) completed three or two experimental sessions, respectively. Mean propulsive velocity (MPVmean) and total number of repetitions (REPtotal) were assessed during a power [3 × 8 at 60% 1RM (one-repetition maximum)] and hypertrophy squat loading (3 sets to failure at 70% 1RM), respectively. Changes in bench press and squat MPV at 60% 1RM in response to the loadings were used as surrogates for nonlocal and local fatigue, respectively. Total blood lactate accumulation (BLAA) and markers of perceived exertion were assessed in each session. No significant differences between any of the MC or OC phases were observed for MPVmean, REPtotal, nonlocal and local fatigue, and markers of perceived exertion (all P > 0.050). A higher intraindividual 17ß-estradiol concentration was significantly associated with a lower MPVmean (P = 0.019). BLAA was significantly higher in the lFP than in the mLP (P = 0.019) and negatively associated with the intraindividual progesterone concentration (P = 0.005). Although 17ß-estradiol may negatively influence the MPV, it appears that fluctuations of both sex hormones across the MC and OC phases are not prominent enough to induce significant or practically relevant changes in the assessed parameters.NEW & NOTEWORTHY Although a high intraindividual 17ß-estradiol concentration was associated with a lower movement velocity, markers of strength performance and surrogates for nonlocal and local fatigue remained unaffected by MC and OC phases. Blood lactate accumulation was significantly reduced in the mLP. Furthermore, our findings suggest that the impact of the MC phases varies greatly among individuals. Individuals with high fluctuations in sex hormone concentrations may experience relevant changes in the assessed parameters.

Getting a Grip on the Undrugged: Targeting ß-Catenin with Fragment-Based Methods.

Aberrant WNT pathway activation, leading to nuclear accumulation of ß-catenin, is a key oncogenic driver event. Mutations in the tumor suppressor gene APC lead to impaired proteasomal degradation of ß-catenin and subsequent nuclear translocation. Restoring cellular degradation of ß-catenin represents a potential therapeutic strategy. Here, we report the fragment-based discovery of a small molecule binder to ß-catenin, including the structural elucidation of the binding mode by X-ray crystallography. The difficulty in drugging ß-catenin was confirmed as the primary screening campaigns identified only few and very weak hits. Iterative virtual and NMR screening techniques were required to discover a compound with sufficient potency to be able to obtain an X-ray co-crystal structure. The binding site is located between armadillo repeats two and three, adjacent to the BCL9 and TCF4 binding sites. Genetic studies show that it is unlikely to be useful for the development of protein-protein interaction inhibitors but structural information and established assays provide a solid basis for a prospective optimization towards ß-catenin proteolysis targeting chimeras (PROTACs) as alternative modality.

Life cycle assessment and sustainable engineering in the context of near net shape grown components: striving towards a sustainable way of future production.

Technical product harvesting (TEPHA) is a newly developing interdisciplinary approach in which bio-based production is investigated from a technical and ecological perspective. Society's demand for ecologically produced and sustainably operable goods is a key driver for the substitution of conventional materials like metals or plastics through bio-based alternatives. Technical product harvesting of near net shape grown components describes the use of suitable biomass for the production of technical products through influencing the natural shape of plants during their growth period. The use of natural materials may show positive effects on the amount of non-renewable resource consumption. This also increases the product recyclability at the end of its life cycle. Furthermore, through the near net shape growth of biomass, production steps can be reduced. As a consequence such approaches may save energy and the needed resources like crude oil, coal or gas. The derived near net shape grown components are not only considered beneficial from an environmental point of view. They can also have mechanical advantages through an intrinsic topology optimization in contrast to common natural materials, which are influenced in their shape after harvesting. In order to prove these benefits a comprehensive, interdisciplinary scientific strategy is needed. Here, both mechanical investigations and life cycle assessment as a method of environmental evaluation are used.

Can Gender-Fair Language Reduce Gender Stereotyping and Discrimination?

Gender-fair language (GFL) aims at reducing gender stereotyping and discrimination. Two principle strategies have been employed to make languages gender-fair and to treat women and men symmetrically: neutralization and feminization. Neutralization is achieved, for example, by replacing male-masculine forms (policeman) with gender-unmarked forms (police officer), whereas feminization relies on the use of feminine forms to make female referents visible (i.e., the applicant… he or she instead of the applicant… he). By integrating research on (1) language structures, (2) language policies, and (3) individual language behavior, we provide a critical review of how GFL contributes to the reduction of gender stereotyping and discrimination. Our review provides a basis for future research and for scientifically based policy-making.

Beyond Sexist Beliefs: How Do People Decide to Use Gender-Inclusive Language?

When people use generic masculine language instead of more gender-inclusive forms, they communicate gender stereotypes and sometimes exclusion of women from certain social roles. Past research related gender-inclusive language use to sexist beliefs and attitudes. Given that this aspect of language use may be transparent to users, it is unclear whether people explicitly act on these beliefs when using gender-exclusive language forms or whether these are more implicit, habitual patterns. In two studies with German-speaking participants, we showed that spontaneous use of gender-inclusive personal nouns is guided by explicitly favorable intentions as well as habitual processes involving past use of such language. Further indicating the joint influence of deliberate and habitual processes, Study 2 revealed that language-use intentions are embedded in explicit sexist ideologies. As anticipated in our decision-making model, the effects of sexist beliefs on language emerged through deliberate mechanisms involving attitudes and intentions.