Changes in ESCRT-III filament geometry drive membrane remodelling and fission in silico

BACKGROUND: ESCRT-III is a membrane remodelling filament with the unique ability to cut membranes from the inside of the membrane neck. It is essential for the final stage of cell division, the formation of vesicles, the release of viruses, and membrane repair. Distinct from other cytoskeletal filaments, ESCRT-III filaments do not consume energy themselves, but work in conjunction with another ATP-consuming complex. Despite rapid progress in describing the cell biology of ESCRT-III, we lack an understanding of the physical mechanisms behind its force production and membrane remodelling. // RESULTS: Here we present a minimal coarse-grained model that captures all the experimentally reported cases of ESCRT-III driven membrane sculpting, including the formation of downward and upward cones and tubules. This model suggests that a change in the geometry of membrane bound ESCRT-III filaments-from a flat spiral to a 3D helix-drives membrane deformation. We then show that such repetitive filament geometry transitions can induce the fission of cargo-containing vesicles. // CONCLUSIONS: Our model provides a general physical mechanism that explains the full range of ESCRT-III-dependent membrane remodelling and scission events observed in cells. This mechanism for filament force production is distinct from the mechanisms described for other cytoskeletal elements discovered so far. The mechanistic principles revealed here suggest new ways of manipulating ESCRT-III-driven processes in cells and could be used to guide the engineering of synthetic membrane-sculpting systems

Association of dentoskeletal morphology with incisor inclination in angle class II patients: a retrospective cephalometric study

Introduction: The purpose of this study was to identify possible dentoskeletal parameters associated with variation of anterior tooth inclination in Angle Class II subdivisions. Methods: Pre-treatment lateral radiographs of 144 Class II patients (68 males, 76 females) aged 9 to 17 years were classified for upper incisor inclination into three groups (proclined, normally inclined, retroclined) homogeneous for gender and skeletal jaw relationship. The effect of age on the 22 cephalometric variables was controlled by covariance analysis. Results: Multivariate analysis of the cephalometric parameters indicated significant inter-group differences. Systematic associations with incisor inclination were revealed using rank correlation: Lower incisor proclination, Wits appraisal and gonial angle significantly decreased (0.04 ≥ p ≥ 0.002), while intercisal angle, mandibular total and corpus length and nasolabial angle increased (0.04 ≥ p ≥ 0.001) with decreasing incisor proclination. Conclusions: Clear-cut classification criteria and control of confounding effects may clarify conflicting previous findings on dentoskeletal differences between Class II subdivisions in the mixed dentition. Only minor dentoskeletal differences appear to be associated with incisor inclination. The increased interincisal and nasolabial angle in Class II division 2 subjects are due to reclination of both upper and lower incisors. Jaw positions and chin prominence are not significantly different between the subdivisions. However, Wits appraisal is decreased in Class II division 2. The increased mandibular length observed in Class II division 2 requires further scrutinization.<br

CD8+ T cells mediate the regenerative PTH effect in hPDL cells via Wnt10b signaling

It was the aim of the present investigation to examine whether the stimulating effect of parathyroid hormone (PTH) on human periodontal ligament (hPDL) cell proliferation and differentiation would be enhanced by hPDL/T-cell interaction involving Wnt10b signaling as a mediating pathway. hPDL cells were cultured from healthy premolar tissues of three adolescent orthodontic patients and exposed to PTH(1-34) in monocultures or co-cultures with CD8(+) T cells. At harvest, proliferation, alkaline phosphatase-specific activity (ALP), and osteocalcin production were determined by immunofluorescence cytochemistry, real-time PCR, biochemical assay, and ELISA. Wnt10b signaling was analyzed by the use of a specific WNT10b neutralizing antibody. PTH(1-34) stimulation of T cells significantly increased Wnt10b expression and production. Wnt10b exposure of hPDL cells enhanced proliferation and differentiation. PDL cells co-cultured with T cells showed a Wnt10b-dependent regulation of proliferation and differentiation parameters. The addition of a Wnt10b-neutralizing Ab to the co-culture medium resulted in a significant inhibition of the PTH(1-34) effect on proliferation, ALP-specific activity, and osteocalcin protein expression. Our findings provide novel insight into the mechanism of action of PTH on hPDL cells and establish the interplay of T cells and hPDL cells via the Wnt10b pathway as a modulating factor for the anabolic properties of the hormone in periodontal regeneration

Housekeeping gene validation for RT-qPCR studies on synovial fibroblasts derived from healthy and osteoarthritic patients with focus on mechanical loading

Selection of appropriate housekeeping genes is essential for the validity of data normalization in reverse transcription quantitative PCR (RT-qPCR). Synovial fibroblasts (SF) play a mediating role in the development and progression of osteoarthritis (OA) pathogenesis, but there is no information on reliable housekeeping genes available. Therefore the goal of this study was to identify a set of reliable housekeeping genes suitable for studies of mechanical loading on SF from healthy and OA patients. Nine genes were evaluated towards expression stability and ranked according their relative stability determined by four different mathematical procedures (geNorm, NormFinder, BestKeeper and comparative ΔCq). We observed that RPLP0 (ribosomal protein, large, P0) and EEF1A1 (eukaryotic translation elongation factor 1 alpha 1) turned out to be the genes with the most stable expression in SF from non-OA or OA patients treated with or without mechanical loading. According to geNorm two genes are sufficient for normalization throughout. Expression of one tested target gene varied considerably, if normalized to different candidate housekeeping genes. Our study provides a tool for accurate and valid housekeeping gene selection in gene expression experiments on SF from healthy and OA patients with and without mechanical loading in consistent with the MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) guidelines and additionally demonstrates the impact of proper housekeeping gene selection on the expression of the gene of interest

The Psychological and Physical Effects of Forests on Human Health: A Systematic Review of Systematic Reviews and Meta-Analyses

BACKGROUND The aim of this systematic review of systematic reviews was to identify, summarise, and synthesise the available evidence of systematic reviews (SRs) and meta-analyses (MAs) on the preventative and therapeutic psychological and physical effects of forest-based interventions. Methods: Both bibliographic databases and grey literature sources were searched for SRs and MAs published until May 2020. Eight databases were searched for relevant articles: MEDLINE, Embase, Web of Science, Cochrane Library, PsycInfo, CiNii, EBSCO, and Scopus. Grey literature was sourced from Google Scholar and other web-based search tools. SRs and MAs that included randomised controlled (RCT), non-randomised controlled (NRCT), and non-controlled trials (NCT) on health-related effects of forest-based interventions were eligible if they had searched at least two databases. The methodological quality of eligible reviews was assessed by AMSTAR-2. Results: We evaluated 11 systematic reviews covering 131 different primary intervention studies, mostly from Asian countries, three of which included supplementary meta-analyses. The quality assessment resulted in moderate confidence in the results of two reviews, low confidence in six, and critically low confidence in three. The results of the eight moderate and low-rated reviews indicated that forest-based interventions are beneficial to the cardiovascular system, immune system, and mental health (in the areas of stress, depression, anxiety, and negative emotions). Evidence for the effectiveness of forest-based interventions on metabolic parameters in adults, the severity of atopic dermatitis in children and adolescents, and social skills and sociality in healthy primary school children was weak. Discussion/Conclusions: Evidence suggests beneficial therapeutic effects of forest-based interventions on hypertension, stress, and mental-health disorders, such as depression and anxiety. Changes in immunological and inflammatory parameters after forest therapy should be verified in bio-geographically native forests. In the future, more attention should be paid to careful planning, implementation, and reporting of primary studies and to systematic reviews on the effects of forest-based interventions

Effects of Kneipp Therapy

Einleitung: Ziel dieser systematischen Übersicht war die Bewertung der verfügbaren Evidenz zur Wirkung der Kneipp-Therapie. Methoden: MEDLINE, Embase, Web of Science, Cochrane-Library und CAMbase wurden nach relevanten Artikeln, veröffentlicht zwischen 2000 und 2019, durchsucht. Graue Literatur wurde über Google Scholar und andere Tools bezogen. Studien mit jeglicher Art von Studiendesign, die die Effekte der Kneipp-Therapie untersuchten, wurden eingeschlossen. Die Qualitätsbewertung erfolgte mittels EPHPP-QAT. Ergebnisse: 25 Quellen, darunter 14 kontrollierte Studien, wurden eingeschlossen. Gemäß EPHPP-QAT wurden 3 Studien \textquotedblstark\textquotedbl, 13 \textquotedblmoderat\textquotedbl und 9 \textquotedblschwach\textquotedbl bewertet. Neun (64%) der kontrollierten Studien berichteten signifikante Verbesserungen nach Kneipp-Therapie im Gruppenvergleich bei chronisch-venöser Insuffizienz, Hypertonie, leichter Herzinsuffizienz, menopausalen Be-schwerden und Schlafstörungen in verschiedenen Patientenkollektiven sowie verbesserte Immunparameter bei gesunden Probanden. Im Hinblick auf Depression und Angst bei Mammakarzinom-Patientinnen mit klimakterischen Beschwerden, Lebensqualität bei Post-Polio-Syndrom, krankheitsbedingten polyneuropathischen Beschwerden und Inzidenz von Erkältungsepisoden bei Kindern konnten keine signifikanten Gruppenunterschiede festgestellt werden. Elf unkontrollierte Studien berichteten Verbesserungen bei allergischen Symptomen, Dyspepsie, Lebensqualität, Herzratenvariabilität, Infekten, Hypertonie, Wohlbefinden, Schmerz und polyneuropathischen Beschwerden. Diskussion/Schlussfolgerung: Die Kneipp-Therapie scheint bei zahlreichen Beschwerdebildern in verschiedenen Patientenkollektiven positive Effekte zu bewirken. Zukünftige Studien sollten noch stärker auf eine methodisch sorgfältige Studienplanung achten (Kontrollgruppen, Randomisierung, adäquate Fallzahlen, Verblindung), um Verzerrungen entgegenzuwirken

Psychological profile and self-administered relaxation in patients with craniofacial pain: a prospective in-office study

Introduction: The objective of this study was to evaluate the psychological profile of craniofacial pain sufferers and the impact of patient subtype classification on the short-time effectiveness of a self-administered relaxation training. Methods: One hundred unselected in-office patients (67% females) suffering from chronic facial pain and/or headache with the presumptive diagnose of temporo-mandibular disorder (TMD) completed a questionnaire battery comprising craniofacial pain perception, somatic complaints, irrational beliefs, and pain behavior and were classified into subtypes using cluster analysis. They underwent a self-administered progressive relaxation training and were re-evaluated for pain perception after 3 months. Results: Pain was mild to moderate in the majority of patients. Symptom domains comprised parafunctional activities, temporo-mandibular pain and dysfunction, fronto-temporal headache, head/neck and neck/back pain. Three patient subtypes were identified regarding symptom/dysfunction level: (i) low burden (mild/moderate), (ii) psychosocial dysfunction (moderate/high), (iii) adaptive coping (moderate/mild). Self-rated adherence to the recommended relaxation training was moderate throughout the sample, but self-rated relief was significantly different between clusters. At follow-up, pain intensity was significantly decreased in all patients, whereas pain-related interference was improved only in dysfunctional and adaptive patients. Improvement of symptom domains varied between clusters and was most comprehensive in adaptive patients. Conclusions: In conclusion, craniofacial pain sufferers can be divided in meaningful subtypes based on their pain perception, irrational beliefs, and pain behaviour. A self-administered relaxation training generally yielded positive effects on pain perception, however the benefit may be greater in patients with more marked symptom impact (both dysfunctional and adaptive).<br

Life cycle assessment and cost evaluation of emerging technologies at early stages: The case of continuous flow synthesis of Rufinamide

In the pharma and fine chemical industries, the development of continuous flow technologies is a process intensification step of primary importance towards the manufacturing of high‐quality products, while reducing the environmental impact and cost of production. The sustainability and profitability of a process can be measured through life cycle Assessment and cost evaluation. However, when applied to emerging technologies, these need to be performed at different stages of the process development in order to limit the uncertainties arising from the scale‐up, and hence providing high‐fidelity projections of environmental impacts and costs at larger scales. The output of the assessment can in fact vary significantly depending on the maturity of the technology and this translates into having different results at commercial scale compared to early estimations. Therefore, in this article, we perform an assessment at two different scales of production, lab and mini‐pilot scale, with the aim of quantifying the uncertainties of the assessment related to the scale‐up, identifying the hotspots of the system, and hence providing guidelines for the further steps of process development. The subject of the assessment is the continuous flow synthesis of Rufinamide. It is the first time that this synthesis is evaluated at pilot‐scale. The results show that low yields in the cycloaddition drastically affect the waste management and the production of precursors, and hence increases environmental impacts and cost of production. This calls for the need of prioritizing the optimization of this synthesis step in order to deploy a green and economically competitive production technology

Floating norms for individualising the ANB angle and the WITS appraisal in orthodontic cephalometric analysis based on guiding variables

Purpose The sagittal skeletal relationship of maxilla and mandible (skeletal class) can generally be determined via lateral cephalograms (ANB angle or Wits appraisal) by comparing measurements to empirical norms based on the respective population mean. However, values differing from these empirical norms also enable a therapeutically desired, normal class I occlusion depending on individual craniofacial pattern, thus requiring floating norms based on guiding variables. As available regression equations consider only few predictor variables and are not up-to-date regarding a contemporary patient collective, the aim of this study was to establish improved and extended regression equations for individualising the ANB angle and Wits appraisal. Methods This retrospective, cross-sectional multicentre study was based on 71 Caucasian male and female subjects of any age with normal dental occlusion. We cephalometrically analysed digitised pretreatment lateral radiographs and performed multiple linear regression analyses to identify suitable skeletal predictor variables for individualising the ANB angle and Wits appraisal. Results Inter- and intrarater reliability tests showed mostly perfect measurement concordance. Both original regression equations by Panagiotidis/Witt and Järvinen could be updated for a contemporary population with new regression coefficients. The equation for individualising the ANB could be further optimised in its prediction reliability by adding the skeletal predictor variables NL-NSL, NSBa, facial axis (Ricketts) and index (Hasund), whereas the recalculated Wits equation could not be further improved by additional guiding variables. Conclusions The improved regression formulae for individualising the ANB angle and Wits appraisal should help to improve the assessment of sagittal skeletal class in clinical orthodontic practice

The role of mechanotransduction versus hypoxia during simulated orthodontic compressive strain—an in vitro study of human periodontal ligament fibroblasts

During orthodontic tooth movement (OTM) mechanical forces trigger pseudo-inflammatory, osteoclastogenic and remodelling processes in the periodontal ligament (PDL) that are mediated by PDL fibroblasts via the expression of various signalling molecules. Thus far, it is unknown whether these processes are mainly induced by mechanical cellular deformation (mechanotransduction) or by concomitant hypoxic conditions via the compression of periodontal blood vessels. Human primary PDL fibroblasts were randomly seeded in conventional six-well cell culture plates with O-2-impermeable polystyrene membranes and in special plates with gas-permeable membranes (Lumox (R), Sarstedt), enabling the experimental separation of mechanotransducive and hypoxic effects that occur concomitantly during OTM. To simulate physiological orthodontic compressive forces, PDL fibroblasts were stimulated mechanically at 2 g.cm(-2) for 48 h after 24 h of pre-incubation. We quantified the cell viability by MTT assay, gene expression by quantitative real-time polymerase chain reaction (RT-qPCR) and protein expression by western blot/enzyme-linked immunosorbent assays (ELISA). In addition, PDL-fibroblast-mediated osteoclastogenesis (TRAP(+) cells) was measured in a 72-h coculture with RAW264.7 cells. The expression of HIF-1 alpha, COX-2, PGE2, VEGF, COL1A2, collagen and ALPL, and the RANKL/OPG ratios at the mRNA/protein levels during PDL-fibroblast-mediated osteoclastogenesis were significantly elevated by mechanical loading irrespective of the oxygen supply, whereas hypoxic conditions had no significant additional effects. The cellular-molecular mediation of OTM by PDL fibroblasts via the expression of various signalling molecules is expected to be predominantly controlled by the application of force (mechanotransduction), whereas hypoxic effects seem to play only a minor role. In the context of OTM, the hypoxic marker HIF-1 alpha does not appear to be primarily stabilized by a reduced O-2 supply but is rather stabilised mechanically