The valve movement response of mussels: a tool in biological monitoring

Biological sensors are becoming more important to monitor the quality of the aquatic environment. In this paper the valve movement response of freshwater (Dreissena polymorpha) and marine (Mytilus edulis) mussels is presented as a tool in monitoring studies. Examples of various methods for data storage and data treatment are presented, elucidating easier operation and lower detection limits. Several applications are mentioned, including an early warning system based on this valve movement response of mussels.     

Ten Years of Practical Experience with the Dreissena-Monitor, a Biological Early Warning System for Continuous Water Quality Monitoring

The Dreissena-Monitor is a biological early warning system for the aquatic environment. It is based on the valve movements of up to 2 × 42 specimens of the zebra mussel Dreissena polymorpha, exposed in a flow-through system of 2 experimental channels connected to the water to be tested. A computer records whether a mussel is open or closed. After an analysis period of 5 min, the percentage of open mussels and the number of valve movements are computed as running averages for each experimental channel. With consideration being given to the normal behaviour of the mussels and the response of the system during several toxicity tests, reliable alarm-thresholds were established which are based on dynamic limits. In addition to these alarm functions (including the automatically safekeeping of a water sample) and the primary measurements, the computer fulfils every function necessary for an early warning system, e.g., complete storage and documentation of all data, validity-check of the primary measurements, and graphical presentation of all results. Together with more than 20 biological early warning systems, the Dreissena-Monitor was tested and evaluated by a joint federal government/federal states project group in Germany. As one of three systems, the Dreissena-Monitor is now recommended for implementation at the measuring stations of the ‘German Commission for the Protection of the Rhine Against Pollution.’ Starting in 1992, the Dreissena-Monitor is now applied at 13 water control stations in Germany (Rhine, Elbe, Danube, several tributaries). The experience of the users at the different stations have revealed that the Dreissena-Monitor complies quite well with the main functional requirements of an early warning system: (1) reliable, unattended operation for at least 1 week, (2) ease of handling, (3) an average of less than 3 h of maintenance per week, and (4) automatic detection of alarm situations within 30 min. In particular, the Dreissena-Monitor detected some water alarms at different rivers that give first clues on the sources of emission.     

A new look at the mechanism of activation and inactivation of voltage-gated ion channels.

Studies on the kinetics of activation and inactivation of the sodium channels of the squid giant axon, on the sodium gating current, and on the properties of the non-inactivating steady-state current, are briefly reviewed. Taken in conjunction with recent evidence on the structure of voltage-gated ion channels, they have led to the development of a series-parallel model of the sodium channel that can be regarded as a modernized version of the Hodgkin-Huxley model, with some novel features. It is suggested that activation results from conformational changes brought about by the four S4 voltage sensors operating in parallel, each of which makes two discrete steps to reach the fully activated state of the channel. There follows a voltage-independent hydration step, and the channel is ready to open. Inactivation is a potential-dependent process involving a third transition of voltage sensor S4d alone, which, rather than bringing a ball and chain blocking group into position to close the channels, serves to switch the system so that it passes from an initial activated mode, in which there is a high probability of arriving at an open state with a brief latency, to a second steady-state mode, in which the probability of opening is very much lower.     

Mg2+ enhances voltage sensor/gate coupling in BK channels

BK (Slo1) potassium channels are activated by millimolar intracellular Mg(2+) as well as micromolar Ca(2+) and membrane depolarization. Mg(2+) and Ca(2+) act in an approximately additive manner at different binding sites to shift the conductance-voltage (G(K)-V) relation, suggesting that these ligands might work through functionally similar but independent mechanisms. However, we find that the mechanism of Mg(2+) action is highly dependent on voltage sensor activation and therefore differs fundamentally from that of Ca(2+). Evidence that Ca(2+) acts independently of voltage sensor activation includes an ability to increase open probability (P(O)) at extreme negative voltages where voltage sensors are in the resting state; 2 microM Ca(2+) increases P(O) more than 15-fold at -120 mV. However 10 mM Mg(2+), which has an effect on the G(K)-V relation similar to 2 microM Ca(2+), has no detectable effect on P(O) when voltage sensors are in the resting state. Gating currents are only slightly altered by Mg(2+) when channels are closed, indicating that Mg(2+) does not act merely to promote voltage sensor activation. Indeed, channel opening is facilitated in a voltage-independent manner by Mg(2+) in a mutant (R210C) whose voltage sensors are constitutively activated. Thus, 10 mM Mg(2+) increases P(O) only when voltage sensors are activated, effectively strengthening the allosteric coupling of voltage sensor activation to channel opening. Increasing Mg(2+) from 10 to 100 mM, to occupy very low affinity binding sites, has additional effects on gating that more closely resemble those of Ca(2+). The effects of Mg(2+) on steady-state activation and I(K) kinetics are discussed in terms of an allosteric gating scheme and the state-dependent interactions between Mg(2+) and voltage sensor that may underlie this mechanism.     

Mode shift of the voltage sensors in Shaker K+ channels is caused by energetic coupling to the pore domain

The voltage sensors of voltage-gated ion channels undergo a conformational change upon depolarization of the membrane that leads to pore opening. This conformational change can be measured as gating currents and is thought to be transferred to the pore domain via an annealing of the covalent link between voltage sensor and pore (S4-S5 linker) and the C terminus of the pore domain (S6). Upon prolonged depolarizations, the voltage dependence of the charge movement shifts to more hyperpolarized potentials. This mode shift had been linked to C-type inactivation but has recently been suggested to be caused by a relaxation of the voltage sensor itself. In this study, we identified two ShakerIR mutations in the S4-S5 linker (I384N) and S6 (F484G) that, when mutated, completely uncouple voltage sensor movement from pore opening. Using these mutants, we show that the pore transfers energy onto the voltage sensor and that uncoupling the pore from the voltage sensor leads the voltage sensors to be activated at more negative potentials. This uncoupling also eliminates the mode shift occurring during prolonged depolarizations, indicating that the pore influences entry into the mode shift. Using voltage-clamp fluorometry, we identified that the slow conformational change of the S4 previously correlated with the mode shift disappears when uncoupling the pore. The effects can be explained by a mechanical load that is imposed upon the voltage sensors by the pore domain and allosterically modulates its conformation. Mode shift is caused by the stabilization of the open state but leads to a conformational change in the voltage sensor.     

Hydrophobic interaction between contiguous residues in the S6 transmembrane segment acts as a stimuli integration node in the BK channel

Large-conductance Ca²⁺- and voltage-activated K⁺ channel (BK) open probability is enhanced by depolarization, increasing Ca²⁺ concentration, or both. These stimuli activate modular voltage and Ca²⁺ sensors that are allosterically coupled to channel gating. Here, we report a point mutation of a phenylalanine (F380A) in the S6 transmembrane helix that, in the absence of internal Ca²⁺, profoundly hinders channel opening while showing only minor effects on the voltage sensor active–resting equilibrium. Interpretation of these results using an allosteric model suggests that the F380A mutation greatly increases the free energy difference between open and closed states and uncouples Ca²⁺ binding from voltage sensor activation and voltage sensor activation from channel opening. However, the presence of a bulky and more hydrophobic amino acid in the F380 position (F380W) increases the intrinsic open–closed equilibrium, weakening the coupling between both sensors with the pore domain. Based on these functional experiments and molecular dynamics simulations, we propose that F380 interacts with another S6 hydrophobic residue (L377) in contiguous subunits. This pair forms a hydrophobic ring important in determining the open–closed equilibrium and, like an integration node, participates in the communication between sensors and between the sensors and pore. Moreover, because of its effects on open probabilities, the F380A mutant can be used for detailed voltage sensor experiments in the presence of permeant cations.     

The circadian rhythms of valve movements in the mussel <Emphasis Type="Italic">Mytilus galloprovincialis</Emphasis>

The long-term (10–30 day) continuous recording of valve movements in the mussel Mytilus galloprovincialis was carried out in the laboratory under nearly natural conditions. Fourier analysis revealed the circadian (close to the diurnal) rhythm of the valve activity, the phase of which is readily shifted by a shift in the beginning of the daytime, since the light regime is one of the main factors determining the circadian rhythm. The circadian rhythm was manifested in the daily dynamics of mussel valve activity: in the daytime, mussels hold their valves closed more often than at night. This behavior may be a protective response, namely the “shadow reflex”: mussels close their valves upon a sudden decrease in illumination, thus protecting themselves from possible predators. Circadian activity can mask a mussel’s response to environmental pollution; therefore, regular valve closure should be taken into account in early warning systems such as “MusselMonitor®” with a correction for the season of the year, time of day, and other factors.     

Review: Automated techniques for monitoring the behaviour and welfare of broilers and laying hens: towards the goal of precision livestock farming

There is increasing public concern about poultry welfare; the quality of animal welfare is closely related to the quality of livestock products and the health of consumers. Good animal welfare promotes the healthy growth of poultry, which can reduce the disease rate and improve the production quality and capacity. As behaviour responses are an important expression of welfare, the study of behaviour is a simple and non-invasive method to assess animal welfare. The use of modern technology offers the possibility to monitor the behaviour of broilers and laying hens in a continuous and automated way. This paper reviews the latest technologies used for monitoring the behaviour of broilers and laying hens under both experimental conditions and commercial applications and discusses the potential of developing a precision livestock farming (PLF) system. The techniques that are presented and discussed include sound analysis, which can be an online tool to automatically monitor poultry behaviour non-invasively at the group level; wireless, wearable sensors with radio-frequency identification devices, which can automatically identify individual chickens, track the location and movement of individuals in real time and quantify some behavioural traits accordingly and image processing technology, which can be considered a direct tool for measuring behaviours, especially activity behaviours and disease early warning. All of these technologies can monitor and analyse poultry behaviour, at the group level or individual level, on commercial farms. However, the popularity and adoption of these technologies has been hampered by the logistics of applying them to thousands and tens of thousands of birds on commercial farms. This review discusses the advantages and disadvantages of these techniques in commercial applications and presents evidence that they provide potential tools to automatically monitor the behaviours of broilers and laying hens on commercial farms. However, there still has a long way to go to develop a PLF system to detect and predict abnormal situations.     

Left ventricular epicardial admittance measurement for detection of acute LV dilation

There are two implanted heart failure warning systems incorporated into biventricular pacemakers/automatic implantable cardiac defibrillators and tested in clinical trials: right heart pressures, and lung conductance measurements. However, both warning systems postdate measures of the earliest indicator of impending heart failure: left ventricular (LV) volume. There are currently no proposed implanted technologies that can perform LV blood volume measurements in humans. We propose to solve this problem by incorporating an admittance measurement system onto currently deployed biventricular and automatic implantable cardiac defibrillator leads. This study will demonstrate that an admittance measurement system can detect LV blood conductance from the epicardial position, despite the current generating and sensing electrodes being in constant motion with the heart, and with dynamic removal of the myocardial component of the returning voltage signal. Specifically, in 11 pigs, it will be demonstrated that 1) a physiological LV blood conductance signal can be derived; 2) LV dilation in response to dose-response intravenous neosynephrine can be detected by blood conductance in a similar fashion to the standard of endocardial crystals when admittance is used, but not when only traditional conductance is used; 3) the physiological impact of acute left anterior descending coronary artery occlusion and resultant LV dilation can be detected by blood conductance, before the anticipated secondary rise in right ventricular systolic pressure; and 4) a pleural effusion simulated by placing saline outside the pericardium does not serve as a source of artifact for blood conductance measurements.     

A fiberoptic sensor system for cardiac monitoring and electrotherapy.

Commercially available cardiac pacemakers and implantable cardioverters/defibrillators (ICD) predominantly use the intracardiac derived electrocardiogram (ECG) for detection of arrhythmias. To achieve an automatic control of the heart frequency in accordance with cardiovascular strain and an improved detection of life-threatening arrhythmias, it is desirable to monitor the heart by an input signal correlated with the hemodynamic state. One possible approach to derive such a signal, is to measure the inotropy (mechanical contraction strength of the heart muscle). For this purpose an optoelectronic measurement system has been designed. The fundamental function of the system has been shown in earlier investigations using an isolated beating pig heart. In this paper further results showing the correlation of the fiberoptic sensor signal with the left ventricular stroke volume are presented. To make the system useful for implantable devices, further improvements with regard to power consumption and signal quality were achieved.     

Atomic constraints between the voltage sensor and the pore domain in a voltage-gated K+ channel of known structure

In voltage-gated K(+) channels (Kv), membrane depolarization promotes a structural reorganization of each of the four voltage sensor domains surrounding the conducting pore, inducing its opening. Although the crystal structure of Kv1.2 provided the first atomic resolution view of a eukaryotic Kv channel, several components of the voltage sensors remain poorly resolved. In particular, the position and orientation of the charged arginine side chains in the S4 transmembrane segments remain controversial. Here we investigate the proximity of S4 and the pore domain in functional Kv1.2 channels in a native membrane environment using electrophysiological analysis of intersubunit histidine metallic bridges formed between the first arginine of S4 (R294) and residues A351 or D352 of the pore domain. We show that histidine pairs are able to bind Zn(2+) or Cd(2+) with high affinity, demonstrating their close physical proximity. The results of molecular dynamics simulations, consistent with electrophysiological data, indicate that the position of the S4 helix in the functional open-activated state could be shifted by approximately 7-8 A and rotated counterclockwise by 37 degrees along its main axis relative to its position observed in the Kv1.2 x-ray structure. A structural model is provided for this conformation. The results further highlight the dynamic and flexible nature of the voltage sensor.     

Position and motions of the S4 helix during opening of the Shaker potassium channel

The four voltage sensors in voltage-gated potassium (Kv) channels activate upon membrane depolarization and open the pore. The location and motion of the voltage-sensing S4 helix during the early activation steps and the final opening transition are unresolved. We studied Zn(2+) bridges between two introduced His residues in Shaker Kv channels: one in the R1 position at the outer end of the S4 helix (R362H), and another in the S5 helix of the pore domain (A419H or F416H). Zn(2+) bridges readily form between R362H and A419H in open channels after the S4 helix has undergone its final motion. In contrast, a distinct bridge forms between R362H and F416H after early S4 activation, but before the final S4 motion. Both bridges form rapidly, providing constraints on the average position of S4 relative to the pore. These results demonstrate that the outer ends of S4 and S5 remain in close proximity during the final opening transition, with the S4 helix translating a significant distance normal to the membrane plane.     

A low-voltage wide-input CMOS comparator for sensor application using back-gate technique

In this paper, two new analog CMOS comparators (Type-I and Type-II) with low-voltage and wide-input capabilities are proposed. The comparator receives two analog inputs and puts out one digital state to identify the larger (or the smaller) of the input variables, which represents an useful operation in data conversion and sensory signal processing. Without using special fabrication technologies, the supply voltage of the circuit is reduced to 1 V. Due to the utilization of CMOS back-gate technique, the input range of the comparators is greatly improved. The comparators are composed of bulk-driven stage and dynamic latch. By using a CMOS n-well technology, the results of HSPICE simulations indicate that the response time of Type-I circuit is 1 micros under 10 mV identified resolution. Type-II comparator achieves 5 mV identified resolution. The input dynamic ranges of the comparators are approximately rail-to-rail.     

A smart device inertial-sensing method for gait analysis

The purpose of this study was to establish and cross-validate a method for analyzing gait patterns determined by the center of mass (COM) through inertial sensors embedded in smart devices. The method employed an extended Kalman filter in conjunction with a quaternion rotation matrix approach to transform accelerations from the object onto the global frame. Derived by double integration, peak-to-trough changes in vertical COM position captured by a motion capture system, inertial measurement unit, and smart device were compared in terms of averaged and individual steps. The inter-rater reliability and levels of agreement for systems were discerned through intraclass correlation coefficients (ICC) and Bland–Altman plots. ICCs corresponding to inter-rater reliability were good-to-excellent for position data (ICCs,.80–.95) and acceleration data (ICCs,.54–.81). Levels of agreements were moderate for position data (LOA, 3.1–19.3%) and poor for acceleration data (LOA, 6.8%–17.8%). The Bland–Altman plots, however, revealed a small systematic error, in which peak-to-trough changes in vertical COM position were underestimated by 2.2 mm; the Kalman filter?s accuracy requires further investigation to minimize this oversight. More importantly, however, the study?s preliminary results indicate that the smart device allows for reliable COM measurements, opening up a cost-effective, user-friendly, and popular solution for remotely monitoring movement. The long-term impact of the smart device method on patient rehabilitation and therapy cannot be underestimated: not only could healthcare expenditures be curbed (smart devices being more affordable than today‘s motion sensors), but a more refined grasp of individual functioning, activity, and participation within everyday life could be attained.     

Xenochironomus canterburyensis (Diptera: Chironomidae) an insectan inquiline commensal of Hyridella menziesi (Mollusca: Lamellibranchia)

Larvae and pupae of the chironomid Xenochironomus canterburyensis (Freeman) are recorded for the first time. They are inquiline commensals of the fresh water mussel Hyridella menziesi (Gray) in Lake Taupo, New Zealand. The chironomid is univoltine and closely dependent on the seasonal growth of the mussel shell for its pupation and adult emergence. The first instar larva is probably a free-swimming, dispersal stage. Second instar larvae appeared in mussels between the outer surface of the mantle and the inner surface of the valve in later summer. They overwintered there as third and fourth instars until spring when they were found at the margin of the valve opposite the inhalent siphon from where they pupated and left the host to emerge at the lake surface. Some mortality of first and second instar larvae occurred as there was an average of 25–5 whole larvae buried in the nacre of each valve of the mussels.     

A single charged voltage sensor is capable of gating the Shaker K+ channel

We sought to determine the contribution of an individual voltage sensor to Shaker''s function. Concatenated heterotetramers of Shaker zH4 Δ(6–46) wild type (wt) in combination with a neutral S4 segment Shaker mutant (mut) with stoichiometries 2wt/2mut and 1wt/3mut were studied and compared with the 4wt concatenated homotetramer. A single charged voltage sensor is sufficient to open Shaker conductance with reduced delay (<1 ms) and at more hyperpolarized voltages compared with 4wt. In addition, the wt-like slow inactivation of 1wt/3mut was almost completely eliminated by mutations T449V-I470C in its single wt subunit, indicating that the subunits bearing a neutral S4 were unable to trigger slow inactivation. Our results strongly suggest that a neutral S4 segment of Shaker''s subunit is voltage insensitive and its voltage sensor is in the activated position (i.e., ready for pore opening), and provide experimental support to the proposed model of independent voltage sensors with a final, almost voltage-independent concerted step.     

A Gating Model for the Archeal Voltage-Dependent K Channel KvAP in DPhPC and POPE:POPG Decane Lipid Bilayers

Voltage-dependent K⁺ (Kv) channels form the basis of the excitability of nerves and muscles. KvAP is a well-characterized archeal Kv channel that has been widely used to investigate many aspects of Kv channel biochemistry, biophysics, and structure. In this study, a minimal kinetic gating model for KvAP function in two different phospholipid decane bilayers is developed. In most aspects, KvAP gating is similar to the well-studied eukaryotic Shaker Kv channel: conformational changes occur within four voltage sensors, followed by pore opening. Unlike the Shaker Kv channel, KvAP possesses an inactivated state that is accessible from the pre-open state of the channel. Changing the lipid composition of the membrane influences multiple gating transitions in the model, but, most dramatically, the rate of recovery from inactivation. Inhibition by the voltage sensor toxin VSTx1 is most easily explained if VSTx1 binds only to the depolarized conformation of the voltage sensor. By delaying the voltage sensor's return to the hyperpolarized conformation, VSTx1 favors the inactivated state of KvAP.     

Thermally Chargeable Solid‐State Supercapacitor

Ubiquitous low‐grade thermal energy, which is typically wasted without use, can be extremely valuable for continuously powering electronic devices such as sensors and wearable electronics. A popular choice for waste heat recovery has been thermoelectric energy conversion, but small output voltage without energy‐storing capability necessitates additional components such as a voltage booster and a capacitor. Here, a novel method of simultaneously generating a large voltage from a temperature gradient and storing electrical energy without losing the benefit of solid‐state no‐moving part devices like conventional thermoelectrics is reported. Thermally driven ion diffusion is used to greatly increase the output voltage (8 mV K^?1) with polystyrene sulfonic acid (PSSH) film. Polyaniline‐coated electrodes containing graphene and carbon nanotube sandwich the PSSH film where thermally induced voltage‐enabled electrochemical reactions, resulting in a charging behavior without an external power supply. With a small temperature difference (5 K) possibly created over wearable energy harvesting devices, the thermally chargeable supercapacitor produce 38 mV with a large areal capacitance (1200 F m^?2). It is anticipated that the attempt with thermally driven ion diffusion behaviors initiates a new research direction in thermal energy harvesting.     

The crystal structure of rabbit phosphoglucose isomerase complexed with D-sorbitol-6-phosphate, an analog of the open chain form of D-glucose-6-phosphate

Phosphoglucose isomerase (PGI) catalyzes the isomerization of D-glucose-6-phosphate (G6P) and D-fructose-6-phosphate (F6P) in glycolysis and gluconeogenesis. Analysis of previously reported X-ray crystal structures of PGI without ligand, with the cyclic form of F6P, or with inhibitors that mimic the cis-enediol intermediate led to proposed mechanisms for the ring opening and isomerization steps in the multistep catalytic mechanism. To help complete our model of the overall mechanism, information is needed about the state of PGI between the ring opening and isomerization steps, in other words, a structure of the enzyme complexed with the open form of a substrate or an analog. Here, we report the crystal structure of rabbit PGI complexed with D-sorbitol-6-phosphate (S6P), an analog of the open chain form of G6P, at 2.0 A resolution. As was seen in the PGI/F6P structure, a helix containing amino acid residues 512-520 is found in the "out" position, which provides sufficient space in the active site for a substrate in its cyclic form and which is probably the location of that helix just after ring opening (or just before ring closure). However, the S6P ligand is in an extended conformation, as was seen previously with ligands that mimic the cis-enediol intermediate. The extended conformation enables the ligand to interact with Glu357, which transfers a proton during the isomerization step. The PGI/S6P structure represents the conformation of the enzyme and substrate between the ring opening (or ring closing) step and the isomerization step and helps to complete the model for PGI's catalytic mechanism.