Interleaved NQR detection using atomic magnetometers.

Interleaved Nuclear Quadrupole Resonance (NQR) detection was conducted on ammonium nitrate and potassium chlorate using two 87Rb magnetometers, where potassium chlorate is measured during the T1 limited recovery time of ammonium nitrate. The multi-pass magnetometers are rapidly matched to the NQR frequencies, 531 kHz and 423 kHz, with the use of a single tuning field. For ease of implementation, a double resonant tank circuit was used for excitation, but could be replaced by a broad-band transmitter. All work was done in an unshielded environment and compared to conventional coil detection. The two magnetometers were sensitive, base noise as low as 2 fT/Hz, and were shown to reduce ambient noise through signal subtraction. When an excitation pulse was introduced, however, residual ringing increased the noise floor; mitigation techniques are discussed. The two detection techniques resulted in comparable Signal-to-Noise Ratio (SNR). Interleaved detection using the atomic magnetometers took half the time of conventional detection and provided localization of the explosives.

Homogeneous fields: Double expansion method, 3D printing/CNC realization, and verification by atomic magnetometry.

In low-field magnetic resonance applications there is often an interest in creating homogeneous magnetic fields over unusual geometries, particularly when quantum magnetometers are involved. In this paper a design method is proposed, where both the surface current and magnetic field are expanded to find current coefficients that cancel out higher order field terms. Two coils are designed using this double expansion methodology: (1) a tuning field for a half-meter-long atomic magnetometer array and (2) a null field for a magnetometer to operate adjacent to an excitation solenoid. The field verification of the former shows the accuracy of CNC milling and the method proposed; a close analysis of the field signature in the latter revealed the limitations of 3D printing for precise scientific applications. Both coils are designed to be fifth-order error systems or better.

RF atomic magnetometer array with over 40 dB interference suppression using electron spin resonance.

An unshielded array of 87Rb atomic magnetometers, operating close to 1 MHz, is used to attenuate interference by 42-48 dB. A sensitivity of 15 fT/Hz to a local source of signal is retained. In addition, a 2D spectroscopic technique, in which the magnetometers are repeatedly pumped and data acquired between pump times, enables a synchronously generated signal to be distinguished from an interfering signal very close in frequency; the timing and signal mimics what would be observed in a magnetic resonance echo train. Combining the interference rejection and the 2D spectroscopy techniques, a 100 fT local signal is differentiated from a 20 pT interference signal operating only 1 Hz away. A phase-encoded reference signal is used to calibrate the magnetometers in real time in the presence of interference. Key to the strong interference rejection is the accurate calibration of the reference signal across the array, obtained through electron spin resonance measurements. This calibration is found to be sensitive to atomic polarization, RF pulse duration, and direction of the excitation. The experimental parameters required for an accurate and robust calibration are discussed.

Chaperone activity of cytosolic small heat shock proteins from wheat.

Small Hsps (sHsps) and the structurally related eye lens alpha-crystallins are ubiquitous stress proteins that exhibit ATP-independent molecular chaperone activity. We studied the chaperone activity of dodecameric wheat TaHsp16.9C-I, a class I cytosolic sHsp from plants and the only eukaryotic sHsp for which a high resolution structure is available, along with the related wheat protein TaHsp17.8C-II, which represents the evolutionarily distinct class II plant cytosolic sHsps. Despite the available structural information on TaHsp16.9C-I, there is minimal data on its chaperone activity, and likewise, data on activity of the class II proteins is very limited. We prepared purified, recombinant TaHsp16.9C-I and TaHsp17.8C-II and find that the class II protein comprises a smaller oligomer than the dodecameric TaHsp16.9C-I, suggesting class II proteins have a distinct mode of oligomer assembly as compared to the class I proteins. Using malate dehydrogenase as a substrate, TaHsp16.9C-I was shown to be a more effective chaperone than TaHsp17.8C-II in preventing heat-induced malate dehydrogenase aggregation. As observed by EM, morphology of sHsp/substrate complexes depended on the sHsp used and on the ratio of sHsp to substrate. Surprisingly, heat-denaturing firefly luciferase did not interact significantly with TaHsp16.9C-I, although it was fully protected by TaHsp17.8C-II. In total the data indicate sHsps show substrate specificity and suggest that N-terminal residues contribute to substrate interactions.

The identity of proteins associated with a small heat shock protein during heat stress in vivo indicates that these chaperones protect a wide range of cellular functions.

The small heat shock proteins (sHSPs) are a ubiquitous class of ATP-independent chaperones believed to prevent irreversible protein aggregation and to facilitate subsequent protein renaturation in cooperation with ATP-dependent chaperones. Although sHSP chaperone activity has been studied extensively in vitro, understanding the mechanism of sHSP function requires identification of proteins that are sHSP substrates in vivo. We have used both immunoprecipitation and affinity chromatography to recover 42 proteins that specifically interact with Synechocystis Hsp16.6 in vivo during heat treatment. These proteins can all be released from Hsp16.6 by the ATP-dependent activity of DnaK and co-chaperones and are heat-labile. Thirteen of the putative substrate proteins were identified by mass spectrometry and reveal the potential for sHSPs to protect cellular functions as diverse as transcription, translation, cell signaling, and secondary metabolism. One of the putative substrates, serine esterase, was purified and tested directly for interaction with purified Hsp16.6. Hsp16.6 effectively formed soluble complexes with serine esterase in a heat-dependent fashion, thereby preventing formation of insoluble serine esterase aggregates. These data offer critical insights into the characteristics of native sHSP substrates and extend and provide in vivo support for the chaperone model of sHSP function.