Measuring macromolecular diffusion using heteronuclear multiple-quantum pulsed-field-gradient NMR

We have previously shown that H-1 pulsed-field-gradient (PFG) NMR spectroscopy provides a facile method for monitoring protein self-association and can be used, albeit with some caveats, to measure the apparent molecular mass of the diffusant [Dingley et al. (1995) J. Biomol. NMR, 6, 321-328]. In this paper we show that, for N-15-labelled proteins, selection of H-1-N-15 multiple-quantum (MQ) coherences in PFG diffusion experiments provides several advantages over monitoring H-1 single-quantum (SQ) magnetization. First, the use of a gradient-selected MQ filter provides a convenient means of suppressing resonances from both the solvent and unlabelled solutes. Second, H-1-N-15 zero-quantum coherence dephases more rapidly than H-1 SQ coherence under the influence of a PFG. This allows the diffusion coefficients of larger proteins to be measured more readily. Alternatively, the gradient length and/or the diffusion delay may be decreased, thereby reducing signal losses from relaxation. In order to extend the size of macromolecules to which these experiments can be applied, we have developed a new MQ PFG diffusion experiment in which the magnetization is stored as longitudinal two-spin order for most of the diffusion period, thus minimizing sensitivity losses due to transverse relaxation and J-coupling evolution.

Validation of ACB in vitro and in vivo as a biomagnetic method for measuring stomach contraction

Background The aim of this study was to validate a biomagnetic method (alternate current biosusceptometry, ACB) for monitoring gastric wall contractions in rats. Methods In vitro data were obtained to establish the relationship between ACB and the strain-gauge (SG) signal amplitude. In vivo experiments were performed in pentobarbital-anesthetized rats with SG and magnetic markers previously implanted under the gastric serosa or after ingestion of magnetic material. Gastric motility was quantified from the tracing amplitudes and frequency profiles obtained by Fast Fourier Transform. Key Results The correlation between in vitro signal amplitudes was strong (R = 0.989). The temporal cross-correlation coefficient between the ACB and SG signal amplitude was higher (P < 0.0001) in the postprandial (88.3 +/- 9.1 V) than in the fasting state (31.0 +/- 16.9 V). Irregular signal profiles, low contraction amplitudes, and smaller signal-to-noise ratios explained the poor correlation between techniques for fasting-state recordings. When a magnetic material was ingested, there was also strong correlation in the frequency and signal amplitude and a small phase-difference between the techniques. The contraction frequencies using ACB were 0.068 +/- 0.007 Hz (postprandial) and 0.058 +/- 0.007 Hz (fasting) (P < 0.002) and those using SG were 0.066 +/- 0.006 Hz (postprandial) and 0.059 +/- 0.008 Hz (fasting) (P < 0.005). Conclusions & Inferences In summary, ACB is reliable for monitoring gastric wall contractions using both implanted and ingested magnetic materials, and may serve as an accurate and sensitive technique for gastrointestinal motility studies.

Tracking the Ovarian Cycle in Black-and-Gold Howlers (Alouatta caraya) by Measuring Fecal Steroids and Observing Vaginal Bleeding

A better understanding of a species` reproductive physiology can help conservation programs to manage primates in the wild and develop assisted reproductive technologies in captivity. We investigated whether measurements of fecal progestin and estrogen metabolites obtained by a radioimmunoassay could be used to monitor the ovarian cycle of Alouatta caraya. We also compared the occurrence of vaginal bleeding with the hormone profiles. We collected fecal samples from 3 adult and 1 subadult captive female over 5 mo and performed vaginal cytology for the adults. The interval between fecal progestin surges in the adult females was 19.11 +/- 2.14 d (n = 18 cycles). Fecal progestin concentrations remained at basal values for 9.83 +/- 2.21 d (n = 18) and rose to elevated values for 9.47 +/- 0.72 d (n = 19). The subadult female showed basal levels of fecal estrogen and progestin concentrations throughout the study, suggesting that our hormone measurements are valid to monitor the ovarian cycle. Bleeding periods coincided with basal levels of fecal estrogens and progestin at intervals of 19.8 +/- 0.9 d and lasted for 4.1 +/- 1.0 d. Although we obtained these data from only 3 individuals, the results indicate that this species likely has a menstrual-type ovarian cycle. These data provide the first endocrine profile for the Alouatta caraya ovarian cycle and are similar to results obtained for other howler species. This similarity is important for comparative studies of howlers, allowing for a better understanding of their reproductive physiology and contributing to a critical information base for managing Alouatta species.

Measuring Bond Strength Between Fiber Post and Root Dentin: A Comparison of Different Tests

Purpose The aim of this study was to evaluate the ability of bond strength tests to accurately measure the bond strength of fiber posts luted into root canals Materials and Methods The test methods studied were hourglass microtensile (HM), push-out (PS), modified push out (MP) and pull out (PL) The evaluated parameters were bond strength values, reliability (using Weibull analysis), failure mode (using confocal microscopy), and stress distribution (using finite element analysis) Forty human intact single rooted and endodontically treated teeth were divided into four groups Each group was assigned one of the test methods The samples in the HM and PS groups were 1 0 +/- 0 1 mm thick, the HM samples were hourglass shaped and the PS samples were disk shaped For the PL and MP groups, each 1 mm dentin slice was luted with a fiber post piece Three dimensional models of each group were made and stress was analyzed based on Von Mises criteria Results PL provided the highest values of bond strength followed by MP both of which also had greater amounts of adhesive failures PS showed the highest frequency of cohesive failures MP showed a more homogeneous stress distribution and a higher Weibull modulus Conclusion The specimen design directly influences the biomechanical behavior of bond strength tests

Nasal mucus transportability in children with cleft palate

Objectives: Children with cleft palate (CP) have a high prevalence of sinusitis. Considering that nasal mucus properties play a pivotal role in the upper airway defense mechanism, the aim of the study was to evaluate nasal mucus transportability and physical properties from children with CP. Setting: Hospital for Rehabilitation of Craniofacial Anomalies, School of Dentistry, University of Sao Paulo, Bauru, SP, Brazil and Laboratory of Experimental Air Pollution, School of Medicine, University of Sao Paulo, Sao Paulo, SP, Brazil. Methods: Nasal mucus samples were collected by nasal aspiration from children with CP and without CP (non-CP). Sneeze clearance (SC) was evaluated by the simulated sneeze machine. In vitro mucus transportability (MCT) by cilia was evaluated by the frog palate preparation. Mucus physical surface properties were assessed by measuring the contact angle (CA). Mucus rheology was determined by means of a magnetic rheometer, and the results were expressed as log G* (vectorial sum of viscosity and elasticity) and tan delta (relationship between viscosity and elasticity) measured at 1 and 100 rad/s. Results: Mucus samples from children with CP had a higher SC than non-CP children (67 +/- 30 and 41 +/- 24 mm, respectively, p < 0.05). Mucus samples from children with CP had a lower CA (24 +/- 16 degrees and 35 +/- 11 degrees, p < 0.05) and a higher tan delta 100 (0.79 +/- 0.24 and 0.51 +/- 0.12, p < 0.05) than non-CP children. There were no significant differences in mucus MCT, log G* 1, tan delta 1 and log G* 100 obtained for CP and non-CP children. Conclusions: Nasal mucus physical properties from children with CP are associated with higher sneeze transportability. The high prevalence of sinusitis in children with CP cannot be explained by changes in mucus physical properties and transportability. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

A titrimetric respirometer measuring the nitrifiable nitrogen in wastewater using in-sensor-experiment

Measurement of nitrifiable nitrogen contained in wastewater by combining the existing respirometric and titrimetric principles is reported. During an in-sensor-experiment using nitrifying activated sludge. both the dissolved oxygen (DO) and pH in the mixed liquor were measured, and the FH was controlled at a set-point through titration of base or acid. A combination of the oxygen uptake rate (OUR), which was obtained from the measured DO signal, and the titration data allowed calculation of the nitrifiable nitrogen and the short-term biological oxygen demand (BOD) of the wastewater sample that was initially added to the sludge. The calculation was based solely on stoichiometric relationships. The approach was preliminarily tested with two types of wastewaters using a prototype sensor. Good correlation was obtained. (C) 2000 Elsevier Science Ltd. All rights reserved.