Tmc1 point mutation affects Ca2+ sensitivity and block by dihydrostreptomycin of the mechanoelectrical transducer current of mouse outer hair cells

The transduction of sound into electrical signals depends on mechanically sensitive ion channels in the stereociliary bundle. The molecular composition of this mechanoelectrical transducer (MET) channel is not yet known. Transmembrane channel-like protein isoforms 1 (TMC1) and 2 (TMC2) have been proposed to form part of the MET channel, although their exact roles are still unclear. Using Beethoven (Tmc1Bth/Bth) mice, which have an M412K point mutation in TMC1 that adds a positive charge, we found that Ca2+ permeability and conductance of the MET channel of outer hair cells (OHCs) were reduced. Tmc1Bth/Bth OHCs were also less sensitive to block by the permeant MET channel blocker dihydrostreptomycin, whether applied extracellularly or intracellularly. These findings suggest that the amino acid that is mutated in Bth is situated at or near the negatively charged binding site for dihydrostreptomycin within the permeation pore of the channel. We also found that the Ca2+ dependence of the operating range of the MET channel was altered by the M412K mutation. Depolarization did not increase the resting open probability of the MET current of Tmc1Bth/Bth OHCs, whereas raising the intracellular concentration of the Ca2+ chelator BAPTA caused smaller increases in resting open probability in Bth mutant OHCs than in wild-type control cells. We propose that these observations can be explained by the reduced Ca2+ permeability of the mutated MET channel indirectly causing the Ca2+ sensor for adaptation, at or near the intracellular face of the MET channel, to become more sensitive to Ca2+ influx as a compensatory mechanism

Hair cell maturation is differentially regulated along the tonotopic axis of the mammalian cochlea

Sound amplification within the mammalian cochlea depends upon specialized hair cells, the outer hair cells (OHCs), which possess both sensory and motile capabilities. In various altricial rodents, OHCs become functionally competent from around postnatal day 7 (P7), before the primary sensory inner hair cells (IHCs), which become competent at about the onset of hearing (P12). The mechanisms responsible for the maturation of OHCs and their synaptic specialization remain poorly understood. We report that spontaneous Ca2+ activity in the immature cochlea, which is generated by CaV1.3 Ca2+ channels, differentially regulates the maturation of hair cells along the cochlea. Under near‐physiological recording conditions we found that, similar to IHCs, immature OHCs elicited spontaneous Ca2+ action potentials (APs), but only during the first few postnatal days. Genetic ablation of these APs in vivo, using CaV1.3−/− mice, prevented the normal developmental acquisition of mature‐like basolateral membrane currents in low‐frequency (apical) hair cells, such as IK,n (carried by KCNQ4 channels), ISK2 and IACh (α9α10nAChRs) in OHCs and IK,n and IK,f (BK channels) in IHCs. Electromotility and prestin expression in OHCs were normal in CaV1.3−/− mice. The maturation of high‐frequency (basal) hair cells was also affected in CaV1.3−/− mice, but to a much lesser extent than apical cells. However, a characteristic feature in CaV1.3−/− mice was the reduced hair cell size irrespective of their cochlear location. We conclude that the development of low‐ and high‐frequency hair cells is differentially regulated during development, with apical cells being more strongly dependent on experience‐independent Ca2+ APs

Investigating the Mechanical Properties of Paperboard Packaging Material for Handling Fresh Produce Under Different Environmental Conditions: Experimental Analysis and Finite Element Modelling

Paper and paperboard are the most widely used packaging materials in the world. The combination of corrugated medium (fluting) and linerboard can be varied to design a corrugated paperboard package in relation to specific mechanical properties of the paper and paperboard. Tensile tests were performed on five different paper grammages (175 g m-2, 200 g m-2, 225 g m-2, 250 g m-2, and 300 g m-2) in the principal directions of the paperboard (machine direction, cross direction and thickness direction) at standard conditions (23°C and 50% relative humidity) and refrigerated transport conditions (0°C and 90% relative humidity). At the same environmental conditions, edgewise compression tests were performed on the corrugated paperboard. Results showed that the mechanical properties of paper and paperboard were affected by the environmental conditions. At the refrigerated transport conditions, the modulus of elasticity strongly decreased in the range of 20 – 53% compared to standard conditions for all the paper grammages. The modulus of elasticity was observed to be higher in the machine direction (MD) than other directions for all the paper grammages. The buckling behaviour of the experimental edgewise compression test of the corrugated paperboard was compared with numerical results. The finite element model of the corrugated paperboard accurately predicted the experimental value of the incipient buckling load with an error of 0.4% and 5.5% at the standard and refrigerated conditions, respectively

Functional Brain Imaging with Multi-Objective Multi-Modal Evolutionary Optimization

Functional brain imaging is a source of spatio-temporal data mining problems. A new framework hybridizing multi-objective and multi-modal optimization is proposed to formalize these data mining problems, and addressed through Evolutionary Computation (EC). The merits of EC for spatio-temporal data mining are demonstrated as the approach facilitates the modelling of the experts' requirements, and flexibly accommodates their changing goals

In silico synchronization reveals regulators of nuclear ruptures in lamin A/C deficient model cells

The nuclear lamina is a critical regulator of nuclear structure and function. Nuclei from laminopathy patient cells experience repetitive disruptions of the nuclear envelope, causing transient intermingling of nuclear and cytoplasmic components. The exact causes and consequences of these events are not fully understood, but their stochastic occurrence complicates in-depth analyses. To resolve this, we have established a method that enables quantitative investigation of spontaneous nuclear ruptures, based on co-expression of a rmly bound nuclear reference marker and a uorescent protein that shuttles between the nucleus and cytoplasm during ruptures. Minimally invasive imaging of both reporters, combined with automated tracking and in silico synchronization of individual rupture events, allowed extracting information on rupture frequency and recovery kinetics. Using this approach, we found that rupture frequency correlates inversely with lamin A/C levels, and can be reduced in genome- edited LMNA knockout cells by blocking actomyosin contractility or inhibiting the acetyl-transferase protein NAT10. Nuclear signal recovery followed a kinetic that is co-determined by the severity of the rupture event, and could be prolonged by knockdown of the ESCRT-III complex component CHMP4B. In conclusion, our approach reveals regulators of nuclear rupture induction and repair, which may have critical roles in disease development

Effect of excise tax increases on cigarette prices in South Africa

The effectiveness of excise tax as a tool for reducing tobacco consumption depends largely on how the tax increases impact retail prices. The study estimates this price relationship in South Africa (2001–2015). The magnitude of the price increase depends on the degree to which the tax increase is passed through to consumers. The degree of passthrough thus determines effectiveness. This paper considers the dynamics of cigarette excise tax passthrough in South Africa. People change their purchasing behavior in response to retail price changes, not in response to excise tax changes.Bill & Melinda Gates Foundation grant number 408254 (IRMA 20177

Using landscape topology to compare continuous metaheuristics: a framework and case study on EDAs and ridge structure

In this paper we extend a previously proposed randomized landscape generator in combination with a comparative experimental methodology to study the behavior of continuous metaheuristic optimization algorithms. In particular, we generate twodimensional landscapes with parameterized, linear ridge structure, and perform pairwise comparisons of algorithms to gain insight into what kind of problems are easy and difficult for one algorithm instance relative to another.We apply thismethodology to investigate the specific issue of explicit dependency modeling in simple continuous estimation of distribution algorithms. Experimental results reveal specific examples of landscapes (with certain identifiable features) where dependency modeling is useful, harmful, or has little impact on mean algorithm performance. Heat maps are used to compare algorithm performance over a large number of landscape instances and algorithm trials. Finally, we perform ameta-search in the landscape parameter space to find landscapes which maximize the performance between algorithms. The results are related to some previous intuition about the behavior of these algorithms, but at the same time lead to new insights into the relationship between dependency modeling in EDAs and the structure of the problem landscape. The landscape generator and overall methodology are quite general and extendable and can be used to examine specific features of other algorithms

Correction accuracy and collateral laxity in open versus closed wedge high tibial osteotomy. A one-year randomised controlled study

In a randomised clinical trial in 50 patients with symptomatic osteoarthritis of the medial compartment of the knee, the clinical results of high tibial osteotomy (HTO) according to the open wedge osteotomy (OWO) and closed wedge osteotomy (CWO) were compared. In both groups locked plate fixation was used. Clinical and radiological assessments were performed preoperatively and after one year. Postoperative hip-knee-ankle (HKA) correction angles were monitored on standing leg X-rays. The effect of HTO on collateral laxity of the knee was measured with a specially designed varus-valgus device. The WOMAC osteoarthritis index, the modified knee society score (KS) and visual analogue scales (VAS) were used to assess symptoms of osteoarthritis, function, pain and patient satisfaction. At one-year follow-up we found accurate corrections in both groups and the planned correction angles were achieved. No loss of correction was observed. Furthermore, the medial collateral laxity and the patellar height significantly decreased after OWO. Significant improvements of WOMAC and KS scores were found in both groups. All patients had significantly less pain and were very satisfied with the results. Surgery time was significantly longer in the CWO group, and complications were more frequent in this group. Both techniques led to good and comparable clinical results. The choice of whether to perform an open or a closed wedge osteotomy may be based on preoperative patellar height or concomitant collateral laxity

Differential activation of killer cells in the circulation and the lung: a study of current smoking status and chronic obstructive pulmonary disease (COPD)

Background:CD8+ T-lymphocytes, natural killer T-like cells (NKT-like cells, CD56+CD3+) and natural killer cells (NK cells, CD56+CD3−) are the three main classes of human killer cells and they are implicated in the pathogenesis of chronic obstructive pulmonary disease (COPD). Activation of these cells can initiate immune responses by virtue of their production of inflammatory cytokines and chemokines that cause lung tissue damage, mucus hypersecretion and emphysema. The objective of the current study was to investigate the activation levels of human killer cells in healthy non-smokers, healthy smokers, ex-smokers with COPD and current smokers with COPD, in both peripheral blood and induced sputum. Methods/Principal Findings:After informed consent, 124 participants were recruited into the study and peripheral blood or induced sputum was taken. The activation states and receptor expression of killer cells were measured by flow cytometry. In peripheral blood, current smokers, regardless of disease state, have the highest proportion of activated CD8+ T-lymphocytes, NKT-like cells and NK cells compared with ex-smokers with COPD and healthy non-smokers. Furthermore, CD8+ T-lymphocyte and NK cell activation is positively correlated with the number of cigarettes currently smoked. Conversely, in induced sputum, the proportion of activated killer cells was related to disease state rather than current smoking status, with current and ex-smokers with COPD having significantly higher rates of activation than healthy smokers and healthy non-smokers. Conclusions: A differential effect in systemic and lung activation of killer cells in COPD is evident. Systemic activation appears to be related to current smoking whereas lung activation is related to the presence or absence of COPD, irrespective of current smoking status. These findings suggest that modulating killer cell activation may be a new target for the treatment of COPD

Evaluating the displacement field of paperboardpackages subjected to compression loading using digital image correlation (DIC)

CITATION: Fadiji, T.; Coetzee, C. J. & Opara, U. L. 2020. Evaluating the displacement field of paperboard packages subjected to compression loading using digital image correlation (DIC). Food and Bioproducts Processing, 123:60-71. doi:10.1016/j.fbp.2020.06.008The original publication is available at https://www.journals.elsevier.com/food-and-bioproducts-processingDigital image correlation (DIC) is a full-field non-contact optical technique for measuring displacements in experimental testing based on correlating several digital images taken during the test, particularly images before and after deformation. Application of DIC cuts across several fields, particularly in experimental solid mechanics; however, its potential application to paperboard packaging has not been fully explored. To preserve fresh horticultural produce during postharvest handling, it is crucial to understand how the packages deform under mechanical loading. In this study, 3D digital image correlation with two cameras and stereovision was used to determine the full-field displacement of corrugated paperboard packaging subjected to compression loading. Strain fields were derived from the displacement fields. Results obtained from the displacement fields showed the initiation and development of the buckling behaviour of the carton panels. The displacement was observed to be largely heterogeneous. The displacement field in the horizontal direction was smaller compared to that of vertical and out-of-plane directions. In addition, the strain variation increased as load increased, which could be a precursor to material failure. The technique proved to be efficient in providing relevant information on the displacement and strain fields at the surface panels of corrugated paperboard packages used for handling horticultural produce. In addition, it offers prospects for improved mechanical design of fresh produce packaging.https://www.sciencedirect.com/science/article/pii/S0960308520304430Publishers versio