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Biomedical subjects

Frederick Sachs

Publications and source records attributed to Frederick Sachs.

7 recordsLinked to original sources
PubMed2002

Microfluidic actuation using electrochemically generated bubbles.

Bubble-based actuation in microfluidic applications is attractive owing to elementary microfabrication requirements. In the present study, the mechanical and chemical characteristics of electrochemically generated bubble valves were studied. By generating electrochemical bubbles as valves directly inside the channel, valves could be closed and opened in milliseconds. Whereas bubble inflation (or valve closing) rate increases with applied voltage, small microfluidic dimensions accelerate bubble deflation rates. It is found that bubbles need not collapse fully to restore full flow, and the channel opens when its hydraulic resistance equals that between the bubble and the wall--a process requiring only milliseconds. Since only picomoles of salt are needed to generate bubbles, pH gradients that are invariably associated with electrochemical reactions were readily suppressed by using a small amount of buffer, as visualized by a pH-sensitive fluorescent dye. A range of common laboratory reagents and electrolytes in varying concentrations, including weak to strong acids and bases, as well as nonaqueous/aqueous mixtures were successfully tested. Using such bubble valves, an eight-way multiplexer was fabricated and tested.

Buffers↗
PubMed2002

Evidence for non-independent gating of P2X2 receptors expressed in Xenopus oocytes.

BACKGROUND: P2X2 receptor is an ATP-activated ion channel which is widely expressed in the nervous system, and mediates synaptic transmission. RESULTS: We recorded currents of P2X2 receptors expressed in Xenopus oocytes from outside-out patches and have found that currents recorded from patches containing a single or multiple P2X2 channels differ in a manner suggesting positive cooperativity. First, the currents from multichannel patches exhibit simultaneous transitions more frequently than predicted from the activity of independent channels. Second, the mean open lifetime at the current level of a single channel in a multichannel burst is about six times longer than the open time of currents from single channel patches, a trend opposite to what is expected of independent channels. These results indicate that the channels have positive cooperativity and that the longer opening is due to a slower closing rate. Third, from kinetic analysis the likelihood of the cooperative model is significantly larger than that of the independent model. Fourth, the open channel noise of currents from patches containing multiple channels is less than half that from a single channel, which is consistent with the channel properties being different when they are active in groups. CONCLUSION: Taken together, our results suggest that P2X2 receptors are non-independent, but interact with positive cooperativity.

Adenosine Triphosphate↗
PubMed2002

High-speed pressure clamp.

We built a high-speed, pneumatic pressure clamp to stimulate patch-clamped membranes mechanically. The key control element is a newly designed differential valve that uses a single, nickel-plated piezoelectric bending element to control both pressure and vacuum. To minimize response time, the valve body was designed with minimum dead volume. The result is improved response time and stability with a threefold decrease in actuation latency. Tight valve clearances minimize the steady-state air flow, permitting us to use small resonant-piston pumps to supply pressure and vacuum. To protect the valve from water contamination in the event of a broken pipette, an optical sensor detects water entering the valve and increases pressure rapidly to clear the system. The open-loop time constant for pressure is 2.5 ms for a 100-mmHg step, and the closed-loop settling time is 500-600 micros. Valve actuation latency is 120 micros. The system performance is illustrated for mechanically induced changes in patch capacitance.

Animals↗
PubMed2002

Solution structure of peptide toxins that block mechanosensitive ion channels.

Mechanosensitive channels (MSCs) play key roles in sensory processing and have been implicated as primary transducers for a variety of cellular responses ranging from osmosensing to gene expression. This paper presents the first structures of any kind known to interact specifically with MSCs. GsMTx-4 and GsMtx-2 are inhibitor cysteine knot peptides isolated from venom of the tarantula, Grammostola spatulata (Suchyna, T. M., Johnson, J. H., Hamer, K., Leykam, J. F., Gage, D. A., Clemo, H. F., Baumgarten, C. M., and Sachs, F. (2000) J. Gen. Physiol. 115, 583-598). Inhibition of cationic MSCs by the higher affinity GsMtx-4 (K(D) approximately 500 nm) reduced cell size in swollen and hypertrophic heart cells, swelling-activated currents in astrocytes, and stretch-induced arrhythmias in the heart. Despite the relatively low affinity, no cross-reactivity has been found with other channels. Using two-dimensional NMR spectroscopy, we determined the solution structure of GsMTx-4 and a lower affinity (GsMTx-2; K(D) approximately 6 microm) peptide from the same venom. The dominant feature of the two structures is a hydrophobic patch, utilizing most of the aromatic residues and surrounded with charged residues. The spatial arrangement of charged residues that are unique to GsMTx-4 and GsMTx-2 may underlie the selectivity of these peptides.

Amino Acid Sequence↗
PubMed2002

Flexoelectricity and elasticity of asymmetric biomembranes.

In view of the well-established charge and dipolar asymmetry of the two leaflets of a native membrane, the theory of flexoelectricity (and curvature elasticity) is extended to take into account this asymmetry using linear and nonlinear forms of the Poisson-Boltzmann equation. The results are discussed with respect to data from atomic force microscopy studies of electromotility in biomembranes.

Biophysical Phenomena↗
PubMedDate not supplied

Dynamic properties of stretch-activated K+ channels in adult rat atrial myocytes.

The effect of mechanical stress on the heart's electrical activity has been termed mechanoelectric feedback. The response to stretch depends upon the magnitude and the waveform of the stimulus, and upon the timing relative to the cardiac cycle. Stretch-activated ion channels (SACs) have been regarded as the most likely candidates for serving as the primary transducers of mechanical stress. We explored the steady state and dynamic responses of single channels in adult rat atrial cells using the patch clamp with a pressure clamp. Surprisingly, we only observed K(+)-selective SACs, probably of the 2P domain family. The channels were weakly outward rectifying with flickery bursts. In cell attached mode, the mean conductance was 74+/-14 and 65+/-16 pS for +60 and -60 mV, respectively (140 mM [K(+)](out), 2mM [Mg(2+)](out) and 0mM [Ca(2+)](out)). The latency of the response to pressure steps was 50-100 ms and the time to peak approximately 400 ms. About half of the channels in cell-attached patches showed adaptation/inactivation where channel activity declined to a plateau of 20-30% of peak in approximately 1s. The time dependent behavior of these SACs is generally consistent with whole-cell currents observed in chick and rat ventricular cells, although the net current was outward rather than inward.

Animals↗