Sensitive detection of type G botulinum neurotoxin through Endopep-MS peptide substrate optimization

Clostridium botulinum produces botulinum neurotoxins (BoNTs) that are one of the most poisonous substances. In order to respond to public health emergencies, there is a need to develop sensitive and specific methods for detecting botulinum toxin in various clinical matrices. Our laboratory has developed a mass spectrometry-based Endopep-MS assay that is able to rapidly detect and differentiate BoNT serotypes A–G by immunoaffinity capture of toxins and detection of unique cleavage products of peptide substrates. To improve the sensitivity of the Endopep-MS assay for the detection of BoNT serotype G, we report here the optimization of synthetic peptide substrates through systematic substitution, deletion, and incorporation of unnatural amino acids. Our data show that the resulting optimized peptides produced a significant improvement (two orders of magnitude) in assay sensitivity and allowed the detection of 0.01 mouseLD₅₀ toxin present in buffer solution.

     

Monitoring of benzylpenicillin decomposition in gastric contents by capillary zone electrophoresis.

A method has been developed to follow the decay of the antibiotic penicillin, specifically penicillin G or benzylpenicillin, in the gastric contents of laboratory rats. Purification by centrifugation and DEAE cellulose treatment of the stomach contents (diluted with pH 9 phosphate-borate buffer) was sufficient to allow the quantification of penicillin by capillary zone electrophoresis. An internal standard was used to minimize the injection error. The loss of activity was greater in fasted animals, as expected from the lower pH of their gastric contents, than in fed rats. The in vivo kinetics of the decomposition of the antibiotic was compared to that obtained in water and in hydrochloric acid solutions.     

High-power picosecond optical parametric oscillator based on periodically poled lithium niobate

A high-power picosecond optical parametric oscillator (OPO) based on a 47-mm periodically poled lithium niobate crystal is described. More than 12 W of total average power-almost 8 W of signal power at 1.85 microm and more than 4 W of idler radiation at 2.5 microm -is simultaneously extracted from less than 18 W of average pump power. The OPO is synchronously pumped by 80-ps (FWHM) cw mode-locked pulses at 1.064 microm , and its output is tunable from 1.7 to 2.84microm . Nearly transform-limited signal pulses are obtained following the introduction of two intracavity etalons.     

Superconductivity in α-phase alloys of Cu,Ag and Au

We have found f.c.c. alloys rich in noble metals to be superconducting. From these data, extrapo lated values for the transition temperatures of Au, Ag and Cu can be obtained.     

Observation of anti-stokes fluorescence cooling in thulium-doped glass

We report the first observation of anti-Stokes fluorescence cooling in a thulium-doped solid with pump excitation at 1.82 &mgr;m相似文献   

Kinetic coefficient of steps at the Si(111) crystal-melt interface from molecular dynamics simulations

Nonequilibrium molecular dynamics simulations are applied to the investigation of step-flow kinetics at crystal-melt interfaces of silicon, modeled with the Stillinger-Weber potential [Phys. Rev. B 31, 5262 (1985)]. Step kinetic coefficients are calculated from crystallization rates of interfaces that are vicinals of the faceted (111) orientation. These vicinal interfaces contain periodic arrays of bilayer steps, and they are observed to crystallize in a step-flow growth mode at undercoolings lower than 40 K. Kinetic coefficients for both [110] and [121] oriented steps are determined for several values of the average step separation, in the range of 7.7-62.4 A. The values of the step kinetic coefficients are shown to be highly isotropic, and are found to increase with increasing step separation until they saturate at step separations larger than approximately 50 A. The largest step kinetic coefficients are found to be in the range of 0.7-0.8 m(sK), values that are more than five times larger than the kinetic coefficient for the rough (100) crystal-melt interface in the same system. The dependence of step mobility on step separation and the relatively large value of the step kinetic coefficient are discussed in terms of available theoretical models for crystal growth kinetics from the melt.     

Early assessment of breast cancer response to neoadjuvant chemotherapy by semi-quantitative analysis of high-temporal resolution DCE-MRI: Preliminary results

Purpose

To evaluate whether semi-quantitative analysis of high temporal resolution dynamic contrast-enhanced MRI (DCE-MRI) acquired early in treatment can predict the response of locally advanced breast cancer (LABC) to neoadjuvant chemotherapy (NAC).

Materials and Methods

As part of an IRB-approved prospective study, 21 patients with LABC provided informed consent and underwent high temporal resolution 3 T DCE-MRI before and after 1 cycle of NAC. Using measurements performed by two radiologists, the following parameters were extracted for lesions at both examinations: lesion size (short and long axes, in both early and late phases of enhancement), radiologist's subjective assessment of lesion enhancement, and percentages of voxels within the lesion demonstrating progressive, plateau, or washout kinetics. The latter data were calculated using two filters, one selecting for voxels enhancing ≥ 50% over baseline and one for voxels enhancing ≥ 100% over baseline. Pretreatment imaging parameters and parameter changes following cycle 1 of NAC were evaluated for their ability to discriminate patients with an eventual pathological complete response (pCR).

Results

All 21 patients completed NAC followed by surgery, with 9 patients achieving a pCR. No pretreatment imaging parameters were predictive of pCR. However, change after cycle 1 of NAC in percentage of voxels demonstrating washout kinetics with a 100% enhancement filter discriminated patients with an eventual pCR with an area under the receiver operating characteristic curve (AUC) of 0.77. Changes in other parameters, including lesion size, did not predict pCR.

Conclusion

Semi-quantitative analysis of high temporal resolution DCE-MRI in patients with LABC can discriminate patients with an eventual pCR after one cycle of NAC.     

Substituent effects on gas-phase acidities of formic acid and its silicon and sulphur derivatives R − M(= X)XH(M = C., Si; X = O,S; R = H,F, CI,OH, NH2 and CH3)

Ab initio molecular orbital methods at the CBS-Q level of theory have been used to study the effect of substituent (F, Cl, NH₂, OH and CH₃) on the gas-phase acidities of formic acid, HCOOH, its silicon and sulphur derivatives R-M(= X)XH(M = C., Si; X = 0, S; R = F, Cl, OH, NH₂ and CH₃). For formic acid and its thio and dithio derivatives the acidity changes upon substitution are irregular and depend on both the type of substituent, position and degree of replacement of oxygen atoms by sulphur atoms. For sila carboxylic acids and their thio and dithio derivatives the calculated acidities regularly increase in the order: R-SiOOH < R-Si(=S)OH ? R-Si(=O)SH < R-SiSSH(R = H, F, Cl, OH, NH₂ and CH₃). The chloro derivatives are the strongest among the acids studied. The highest gas phase acidity (1277.6 kJmol^?1) has been calculated for ClC(=S)OH.