Extracting, quantifying, and comparing dynamical and biomechanical properties of living matter through single particle tracking.

A panoply of new tools for tracking single particles and molecules has led to an explosion of experimental data, leading to novel insights into physical properties of living matter governing cellular development and function, health and disease. In this Perspective, we present tools to investigate the dynamics and mechanics of living systems from the molecular to cellular scale via single-particle techniques. In particular, we focus on methods to measure, interpret, and analyse complex data sets that are associated with forces, materials properties, transport, and emergent organisation phenomena within biological and soft-matter systems. Current approaches, challenges, and existing solutions in the associated fields are outlined in order to support the growing community of researchers at the interface of physics and the life sciences. Each section focuses not only on the general physical principles and the potential for understanding living matter, but also on details of practical data extraction and analysis, discussing limitations, interpretation, and comparison across different experimental realisations and theoretical frameworks. Particularly relevant results are introduced as examples. While this Perspective describes living matter from a physical perspective, highlighting experimental and theoretical physics techniques relevant for such systems, it is also meant to serve as a solid starting point for researchers in the life sciences interested in the implementation of biophysical methods.

Foregut organ progenitors and their niche display distinct viscoelastic properties in vivo during early morphogenesis stages.

Material properties of living matter play an important role for biological function and development. Yet, quantification of material properties of internal organs in vivo, without causing physiological damage, remains challenging. Here, we present a non-invasive approach based on modified optical tweezers for quantifying sub-cellular material properties deep inside living zebrafish embryos. Material properties of cells within the foregut region are quantified as deep as 150 µm into the biological tissue through measurements of the positions of an inert tracer. This yields an exponent, α, which characterizes the scaling behavior of the positional power spectra and the complex shear moduli. The measurements demonstrate differential mechanical properties: at the time when the developing organs undergo substantial displacements during morphogenesis, gut progenitors are more elastic (α = 0.57 ± 0.07) than the neighboring yolk (α = 0.73 ± 0.08), liver (α = 0.66 ± 0.06) and two mesodermal (α = 0.68 ± 0.06, α = 0.64 ± 0.06) progenitor cell populations. The higher elasticity of gut progenitors correlates with an increased cellular concentration of microtubules. The results infer a role of material properties during morphogenesis and the approach paves the way for quantitative material investigations in vivo of embryos, explants, or organoids.

Changes in Cell Morphology and Actin Organization in Embryonic Stem Cells Cultured under Different Conditions.

The cellular cytoskeleton provides the cell with a mechanical rigidity that allows mechanical interaction between cells and the extracellular environment. The actin structure plays a key role in mechanical events such as motility or the establishment of cell polarity. From the earliest stages of development, as represented by the ex vivo expansion of naïve embryonic stem cells (ESCs), the critical mechanical role of the actin structure is becoming recognized as a vital cue for correct segregation and lineage control of cells and as a regulatory structure that controls several transcription factors. Naïve ESCs have a characteristic morphology, and the ultrastructure that underlies this condition remains to be further investigated. Here, we investigate the 3D actin cytoskeleton of naïve mouse ESCs using super-resolution optical reconstruction microscopy (STORM). We investigate the morphological, cytoskeletal, and mechanical changes in cells cultured in 2i or Serum/LIF media reflecting, respectively, a homogeneous preimplantation cell state and a state that is closer to embarking on differentiation. STORM imaging showed that the peripheral actin structure undergoes a dramatic change between the two culturing conditions. We also detected micro-rheological differences in the cell periphery between the cells cultured in these two media correlating well with the observed nano-architecture of the ESCs in the two different culture conditions. These results pave the way for linking physical properties and cytoskeletal architecture to cell morphology during early development.

Septoplastie chez l'enfant: indications et résultats

Introduction : Pendant plusieurs décennies, la pratique de septoplastie chez l'enfant était proscrite par la majorité des chirurgiens par crainte d'altération de la croissance craniofaciale. Actuellement, la septoplastie est de plus en plus indiquée chez l'enfant avec de bons résultats fonctionnels. Le but de ce travail est de révéler les particularités de la septoplastie chez l'enfant, de comparer les différentes techniques opératoires proposées etd'évaluer les résultats obtenus.Méthodes : Notre étude est rétrospective monocentrique transversale portant sur 34 enfants opérés de septoplastie sur une période de 15 ans [2000-2014].Résultats : L'âge moyen de nos patients était de 13 ans avec un sex-ratio de 5,8. Les antécédents d'adénoïdectomie ont été notés dans 9% des cas associés à une amygdalectomie dans 6% des cas. La déviation septale était posttraumatique dans 59% des cas.Elle était de siège antérieur dans 91% des cas. A côté de la déviation de la cloison nasale, nous avons noté une déviation de la pyramide nasale dans 32% des cas et un polype de Killian associé dans 3% des cas. L'indication opératoire était posée sur l'évaluation subjective de la gêne respiratoire et du degré de la déformation nasale. Une obstruction nasale sévère avec réduction importante du flux aérien a été retrouvée dans 73% des cas. Tous nos patients ont été opérés sous anesthésie générale. L'intervention était pratiquée selon la technique de Cottle dans 88% des cas et par voie endoscopique dans 12% des cas. Elle a consisté à une reposition simple de la cloison nasale dans 3% des cas, une dépose repose de la cloison dans 41% des cas, une résection sous muqueuse du cartilage dans 56% des cas. On n'a pas noté de complications majeurs post opératoires.Après un recul moyen de 22 mois, nous avons un bon résultat fonctionnel dans 88% des cas. La persistance de l'obstruction nasale a été notée dans 12% des cas et était en rapport avec la déviation septale dans 6% des cas. Une reprise chirurgicale était indiquée dans 6% des cas.Conclusion :La septoplastie de l'enfant doit être un geste conservateur limité à la zone pathologique. Le suivi post opératoire est clinique et prolongé

Dynamics of membrane nanotubes coated with I-BAR.

Membrane deformation is a necessary step in a number of cellular processes such as filopodia and invadopodia formation and has been shown to involve membrane shaping proteins containing membrane binding domains from the IRSp53-MIM protein family. In reconstituted membranes the membrane shaping domains can efficiently deform negatively charged membranes into tubules without any other proteins present. Here, we show that the IM domain (also called I-BAR domain) from the protein ABBA, forms semi-flexible nanotubes protruding into Giant Unilamellar lipid Vesicles (GUVs). By simultaneous quantification of tube intensity and tubular shape we find both the diameter and stiffness of the nanotubes. I-BAR decorated tubes were quantified to have a diameter of ~50 nm and exhibit no stiffening relative to protein free tubes of the same diameter. At high protein density the tubes are immobile whereas at lower density the tubes diffuse freely on the surface of the GUV. Bleaching experiments of the fluorescently tagged I-BAR confirmed that the mobility of the tubes correlates with the mobility of the I-BAR on the GUV membrane. Finally, at low density of I-BAR the protein upconcentrates within tubes protruding into the GUVs. This implies that I-BAR exhibits strong preference for negatively curved membranes.

Marginal and apical bone stability after staged sinus floor augmentation using bone condensing implants with variable-thread design: a two-dimensional analysis.

Studies on the vertical stability of augmented bone after sinus lifting differ substantially. In addition, long healing periods are usually advocated prior to implant installation. The purpose of this case series study was to evaluate the changes in bone height after sinus lifting with a bovine-derived xenograft and to evaluate the clinical outcome of bone condensing implants installed after a short healing period. Patients treated during the years 2010-2013 were re-examined using peri-apical radiographs to evaluate the changes in augmented bone height (BH) and marginal bone loss (BL). Fifty-seven of 70 eligible subjects (28 male and 29 female, mean age 56 years) attended for reassessment. Data were available for 53 sinus lifts and 105 implants installed after a mean healing period of 4.6±1.5 months. Implant survival was 99% after a mean time in function of 19±9 months. Baseline BH, BH at implant placement, and final BH were on average 3.87±1.74mm, 13.75±2.12mm, and 13.11±2.12mm, respectively (P<0.001). Mean BL was 0.51±0.65mm. Only limited resorption is to be expected after sinus lifting in the short term. A bone condensing implant can be used in the early healing phase with successful outcomes in terms of implant survival and bone adaptation.

Fluorescent quantification of size and lamellarity of membrane nanotubes.

Membrane nanotubes, ubiquitous in cellular systems, adopt a spectrum of curvatures and shapes that are dictated by their intrinsic physical characteristics as well as their interactions with the local cellular environment. A high bending flexibility is needed in the crowded cytoplasm where tubes often need to bend significantly in the axial direction at sub-micron length scales. We find the stiffness of spontaneously formed membrane nanotubes by measuring the persistence length of reconstituted membrane nanotubes freely suspended in solution and imaged by fluorescence microscopy. By quantifying the tube diameter we demonstrate for the first time that the persistence length scales linearly with radius. Although most tubes are uni-lamellar, the predicted linear scaling between tube radius and persistence length allows us to identify tubes that spontaneously form as multilamellar structures upon hydration. We provide the first experimental evidence that illumination of lipid fluorophores can have a profound effect on the lipid bilayer which we sensitively detect as a continuous change in the tube persistence length with time. The novel assay and methodology here presented has potential for quantification of the structural reinforcement of membrane tubes by scaffolding proteins.

Dental implants placed by undergraduate students: clinical outcomes and patients'/students' perceptions.

INTRODUCTION: Implant dentistry in undergraduate education is predominantly theoretical or prosthetics oriented. Clinical experience with implant surgery could provide students a better understanding of alternatives for tooth replacements. This study describes an implant dentistry programme for undergraduate students, which included surgical placement of implants. The study presents the clinical outcomes of the programme, patients' satisfaction and students' attitudes/perceptions. It reflects on barriers and problems encountered during implementation and provides suggestions for other institutions. MATERIALS AND METHODS: Thirty-six students placed one implant each for a single tooth replacement after careful radiographic assessment and pre-surgical planning. One-stage surgery was performed under one-to-one supervision. Crowns were cemented on individual abutments 3-6 months later. Crestal bone loss was assessed radiographically immediately after surgery, at crown placement and after 1 year of loading. Questionnaires were used to investigate patients' perspectives and students' opinions towards the programme, as well as their perceived level of competence. RESULTS: Thirty-six implants were placed in 27 patients; two (5.6%) failed prior to loading; mean bone loss from time of surgery to crown placement was 1.41 mm and remained unchanged thereafter, reflecting implant success. Overall, patients were satisfied and the majority would repeat the treatment by a student. The students thought it was a valuable experience, although they realised that additional education is necessary to perform implant surgery without supervision. CONCLUSION: Implant placement by undergraduate students resulted in acceptable clinical outcome parameters, patient satisfaction and positive student perceptions. These findings support the further development of clinical implant education in undergraduate dental curricula.

FBAR syndapin 1 recognizes and stabilizes highly curved tubular membranes in a concentration dependent manner.

Syndapin 1 FBAR, a member of the Bin-amphiphysin-Rvs (BAR) domain protein family, is known to induce membrane curvature and is an essential component in biological processes like endocytosis and formation and growth of neurites. We quantify the curvature sensing of FBAR on reconstituted porcine brain lipid vesicles and show that it senses membrane curvature at low density whereas it induces and reinforces tube stiffness at higher density. FBAR strongly up-concentrates on the high curvature tubes pulled out of Giant Unilamellar lipid Vesicles (GUVs), this sorting behavior is strongly amplified at low protein densities. Interestingly, FBAR from syndapin 1 has a large affinity for tubular membranes with curvatures larger than its own intrinsic concave curvature. Finally, we studied the effect of FBAR on membrane relaxation kinetics with high temporal resolution and found that the protein increases relaxation time of the tube holding force in a density-dependent fashion.

Drug-related hospitalization at a tertiary teaching center in Lebanon: incidence, associations, and relation to self-medicating behavior.

OBJECTIVE: In Lebanon there is very limited restriction on drug use. Accordingly, self-medication is highly prevalent. This study examined the influence of these factors on the development of drug-related illnesses that lead to hospitalization. METHODS: Patients admitted to the medical and pediatric wards of a tertiary teaching center in Beirut, Lebanon, over a period of 6 months were interviewed and their charts were reviewed. Admissions attributable to adverse drug reactions or therapeutic failures were identified and characterized with respect to demographic factors, medical history, drug intake, and self-medicating behavior. The influence of these variables on the development of drug-related illnesses was examined by logistic regression. RESULTS: Of 1745 adults and 457 children, there were 177 (10.2%) and 36 (7.9%) drug-related illnesses, respectively. Adverse drug reactions accounted for 7.0% and 5.7% and therapeutic failures for 3.2% and 2.2% of adult and pediatric admissions, respectively. Self-medication was commonly practiced (52.6% of adults and 41.6% of children). Logistic regression analysis revealed that female sex increased the risk of adverse drug reaction in adults, whereas self-medication decreased the risk. In children, the risk of adverse drug reaction was increased in lower socioeconomic groups, whereas the risk of therapeutic failure was increased by a positive history of atopy or drug reaction. CONCLUSIONS: These results provide the first detailed analysis of the problem of drug-related illnesses in a developing country and identify a number of related or risk factors. Despite the lack of regulation of drug dispensing and the unchecked access to drugs in Lebanon, the incidence of drug-related illnesses is not different from that in Western nations. This finding may have relevance to policies of drug regulation in other countries.

Expression of TNF and the 55-kDa TNF receptor in epidermis, oral mucosa, lichen planus and squamous cell carcinoma.

UNLABELLED: TNF has diverse biological effects including a role in the immune response and growth regulatory effects. OBJECTIVES: The aim of this study was to determine the nature and location of cells able to synthesise and respond to TNF in oral mucosa in health and disease. MATERIALS AND METHODS: The location of immunoreactive TNF and 55-kDa and 75-kDa TNF receptors was demonstrated using immunohistology. We also used RT-PCR to help determine the sites of synthesis of this cytokine in oral mucosa. RESULTS: Only occasional TNF-positive cells were detected in normal epidermis and oral mucosa. However, this cytokine was found throughout the epithelia in oral lichen planus, leukoplakia and squamous cell carcinoma (SCC). The possible cellular sources and biological effects of TNF in these disease processes is discussed. The 55-kDa TNFR was expressed at the cell membrane throughout the epidermis, but confined to the lower cell layers on oral mucosa. Ten of 22 SCC did not express this receptor. A significant correlation was observed between the velocity of tumour growth and absence of TNF receptors. CONCLUSIONS: The growth inhibitory and immunoregulatory effects of TNF may be modulated by changes in receptor expression and alterations in synthesis of this cytokine in a subgroup of oral SCC.