Calcium Wave Promotes Cell Extrusion

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How do we overcome abrupt degradation of marine ecosystems and meet the challenge of heat waves and climate extremes?

Extreme heat wave events are now causing ecosystem degradation across marine ecosystems. The consequences of this heat‐induced damage range from the rapid loss of habitat‐forming organisms, through to a reduction in the services that ecosystems support, and ultimately to impacts on human health and society. How we tackle the sudden emergence of ecosystem‐wide degradation has not yet been addressed in the context of marine heat waves. An examination of recent marine heat waves from around Australia points to the potential important role that respite or refuge from environmental extremes can play in enabling organismal survival. However, most ecological interventions are being devised with a target of mid to late‐century implementation, at which time many of the ecosystems, that the interventions are targeted towards, will have already undergone repeated and widespread heat wave induced degradation. Here, our assessment of the merits of proposed ecological interventions, across a spectrum of approaches, to counter marine environmental extremes, reveals a lack preparedness to counter the effects of extreme conditions on marine ecosystems. The ecological influence of these extremes are projected to continue to impact marine ecosystems in the coming years, long before these interventions can be developed. Our assessment reveals that approaches which are technologically ready and likely to be socially acceptable are locally deployable only, whereas those which are scalable—for example to features as large as major reef systems—are not close to being testable, and are unlikely to obtain social licence for deployment. Knowledge of the environmental timescales for survival of extremes, via respite or refuge, inferred from field observations will help test such intervention tools. The growing frequency of extreme events such as marine heat waves increases the urgency to consider mitigation and intervention tools that support organismal and ecosystem survival in the immediate future, while global climate mitigation and/or intervention are formulated.     

Detection of Cryptomeria japonica roots with ground penetrating radar

Abstract

Coarse tree roots, which are responsible for most root carbon storage, are usually measured by destructive methods such as excavation and coring. Ground penetrating radar (GPR) is a non-destructive tool that could be used to detect coarse roots in forest soils. In this study, we examined whether the roots of Cryptomeria japonica, a major plantation species in Japan, can be detected with GPR. We also looked for factors that impact the analysis and detection of roots. Roots and wooden dowels of C. japonica were buried 30 cm deep in sandy granite soil. From GPR measurements with a 900 MHz antenna, the distribution and diameter of samples in several transects were recorded. The buried roots were detected clearly and could be distinguished at diameters of 1.1–5.2 cm. There were significant positive relationships between root diameter and parameters extracted from the resultant GPR waveform. The difference in water content between roots and soil is a crucial factor impacting the ability to detect roots with GPR. We conclude that GPR can be used as a non-destructive tool, but further investigation is needed to determine optimal conditions (e.g. water content) and analytical methods for using GPR to examine roots in forest sites.     

Fabrication,Operation and Flow Visualization in Surface-acoustic-wave-driven Acoustic-counterflow Microfluidics

Surface acoustic waves (SAWs) can be used to drive liquids in portable microfluidic chips via the acoustic counterflow phenomenon. In this video we present the fabrication protocol for a multilayered SAW acoustic counterflow device. The device is fabricated starting from a lithium niobate (LN) substrate onto which two interdigital transducers (IDTs) and appropriate markers are patterned. A polydimethylsiloxane (PDMS) channel cast on an SU8 master mold is finally bonded on the patterned substrate. Following the fabrication procedure, we show the techniques that allow the characterization and operation of the acoustic counterflow device in order to pump fluids through the PDMS channel grid. We finally present the procedure to visualize liquid flow in the channels. The protocol is used to show on-chip fluid pumping under different flow regimes such as laminar flow and more complicated dynamics characterized by vortices and particle accumulation domains.     

Adverse responses of Cabera pusaria caterpillars to high dietary manganese concentration

The adaptation of insects to environmental changes can constitute a crucial factor in their development and activity. The response of Cabera pusaria L. (Lepidoptera: Geometridae) caterpillars to high manganese (Mn) concentrations in the diet was studied. Birch leaves were treated by dipping in MnCl₂·4H₂O solutions, thereby achieving Mn contents of 370 (T0), 695 (T1), 3 198 (T2), and 6 302 mg kg⁻¹ (T3). The reactions were determined by observing caterpillar mortality, development time, food consumption, and pupal weight. Manganese concentrations in larval excrement, pupae, and food were determined. Manganese in the diet at unnaturally increased concentrations caused great stress for caterpillars. All individuals in the treatment with the highest Mn concentration (T3) died during rearing and successful pupation occurred in only four individuals in T2. Even in the case of caterpillars from T1 (twofold higher than T0) a negative reaction (increased food consumption and prolonged development) was recorded. We also determined significantly increased Mn concentration in pupae from T1 (T2 and T3 were not included in this evaluation due to mortality) and excrement (T1‒T3) compared with T0 having a natural Mn concentration. Caterpillars were seen to eliminate negatively acting dietary Mn by its translocation to excrement. However, the highest mortality rate in T2 and T3 and negative reactions of individuals in T1 very likely demonstrate energy insufficiency and the high energy requirements of Mn elimination mechanisms.     

Very high density of CHO cells in perfusion by ATF or TFF in WAVE bioreactor™. Part I. Effect of the cell density on the process

High cell density perfusion process of antibody producing CHO cells was developed in disposable WAVE Bioreactor? using external hollow fiber filter as cell separation device. Both “classical” tangential flow filtration (TFF) and alternating tangential flow system (ATF) equipment were used and compared. Consistency of both TFF‐ and ATF‐based cultures was shown at 20–35 × 10⁶ cells/mL density stabilized by cell bleeds. To minimize the nutrients deprivation and by‐product accumulation, a perfusion rate correlated to the cell density was applied. The cells were maintained by cell bleeds at density 0.9–1.3 × 10⁸ cells/mL in growing state and at high viability for more than 2 weeks. Finally, with the present settings, maximal cell densities of 2.14 × 10⁸ cells/mL, achieved for the first time in a wave‐induced bioreactor, and 1.32 × 10⁸ cells/mL were reached using TFF and ATF systems, respectively. Using TFF, the cell density was limited by the membrane capacity for the encountered high viscosity and by the pCO₂ level. Using ATF, the cell density was limited by the vacuum capacity failing to pull the highly viscous fluid. Thus, the TFF system allowed reaching higher cell densities. The TFF inlet pressure was highly correlated to the viscosity leading to the development of a model of this pressure, which is a useful tool for hollow fiber design of TFF and ATF. At very high cell density, the viscosity introduced physical limitations. This led us to recommend cell densities under 1.46 × 10⁸ cell/mL based on the analysis of the theoretical distance between the cells for the present cell line. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:754–767, 2013     

The shear modulus of lower-leg muscles correlates to intramuscular pressure

Shear wave elastography (SWE) is emerging as an innovative tool to evaluate muscle properties and function. It has been shown to correlate with both passive and active muscle forces, and is sensitive to physiological processes and pathological conditions. Similarly, intramuscular pressure (IMP) is an important parameter that changes with passive and active muscle contraction, body position, exercise, blood pressure, and several pathologies. Therefore, the objective of this study was to quantify the dependency of shear modulus within the lower-leg muscles on IMP in healthy individuals. Nineteen healthy individuals (age: Mean age ± SD, 23.84 ± 6.64 years) were recruited. Shear modulus was measured using ultrasound SWE on the tibialis anterior (TA) and peroneus longus (PL) muscles using pressure cuff inflation around the thigh at 40 mmHg, 80 mmHg, and 120 mmHg. Changes in IMP were verified using a catheter connected to a blood pressure monitor. It was found that IMP was correlated to TA and PL shear modulus (spearman's rank correlation = 0.99 and 0.99, respectively). Applying a gradual increase of cuff pressure from 0 to 120 mmHg increased the shear modulus of the TA and PL muscles from 15.83 (2.46) kPa to 21.88 (4.33) kPa and from 9.64 (1.97) kPa to 12.88 (5.99) kPa, respectively. These results demonstrate that changes of muscle mechanical properties are dependent on IMP. This observation is important to improve interpretation of ultrasound elastograms and to potentially use it as a biomarker for more accurate diagnosis of pathologies related to increased IMP.     

Abdominal girth as an index of muscle tension during abdominal hollowing: Selecting the optimal training intensity for the transversus abdominis muscle

The abdominal hollowing technique is used for training the transversus abdominis (TrA). However, the optimal intensity of hollowing is still unclear. The objective of the present study is to verify the validity of estimating the tension of the TrA by measuring the girth of the abdomen with a tape and to determine the optimum intensity of hollowing to effectively train the TrA. Sixteen healthy males performed hollowing with an intensity of 0%, 25%, 50%, 75%, and 100%, estimated from the girth of the abdomen. The shear elastic modulus was measured for the rectus abdominis (RA), external oblique (EO), internal oblique (IO), and TrA at all intensities via ultrasonic shear wave elastography. The shear elastic modulus was considered as the index of the tension of the abdominal muscles at each intensity, and the ratio of the TrA to RA, EO, and IO respectively was calculated as the index of TrA selectivity. As the intensity of hollowing increased, the girth of abdomen decreased and tension of all the four muscles increased. The ratio of TrA to the RA, EO, and IO did not exhibit a significant variation among hollowing intensities of 25% to 100%. It is rational to estimate the tension of the TrA by measuring the girth of the abdomen. Moreover, considering both TrA contraction intensity and selectivity, abdominal hollowing performed at maximum intensity was effective for the maximum contraction training of the TrA.