RESUMEN
Non covalent grafting of proteins on affinity phases is a very common approach for isolation, purification and re-concentration of tagged proteins. Many biophysical studies are conducted on these grafted proteins (surface plasmon resonance, quartz crystal microbalance, etc.) showing that the integrity and function of the protein is usually maintained. However, NMR studies of such samples were not undertaken so far, due to the broadening observed on this kind of heterogeneous samples. We present here the use of the HR-MAS technology to obtain 2D NMR spectra of the MAGI-1 PDZ2/6 protein domain, C13-labeled, tagged with a His-tag and grafted on a Nickel affinity resin. We optimized the C13 Methyl SOFAST HMQC experiment allowing important gains in terms of signal-to-noise. The gain comes from the gathering of proton magnetization from the resin material to the protein under study. Several methyl signals from the unstructured C-terminal tail, which is involved in the binding of the PDZ domain to C-terminal peptides of its partners, were observed and measured. The interaction of the bound PDZ domain with cognate peptides was monitored using <500µg of protein sample. A response proportional to the peptide Kd is obtained, indicating that the method can be used to rapidly and efficiently monitor protein-ligand interactions.
Asunto(s)
Espectroscopía de Resonancia Magnética/métodos , Proteínas/química , Proteínas/metabolismo , Moléculas de Adhesión Celular Neuronal/química , Moléculas de Adhesión Celular Neuronal/metabolismo , Ligandos , Péptidos/química , Péptidos/metabolismo , Estructura Terciaria de ProteínaRESUMEN
We have developed a database and an analysis program (NoiseScan) for noise-induced hearing loss (NIHL). The exposure data are based on the evaluation of the noise immission level, which includes duration, frequency content, and the use of, and the attenuation performance of, hearing protectors. The input data can handle an unlimited number of exposure periods. If the noise exposure level is not known, the program lists noise levels of comparable work places, and thus provides an estimate of exposure. Confounding medical factors that may contribute to NIHL, such as elevated serum cholesterol level, hypertension, and extensive use of pain killers, are collected. Combined exposure to agents that clearly contribute to NIHL, such as hand-arm vibration, tobacco smoking, use of aminoglycosides and exposure to solvents are also assessed. An unlimited number of audiograms can be stored, and all the data can be completed and edited following collection. The program gives the predicted hearing loss according to the ISO 1999 model based on total exposure. At present, our NoiseScan program (under continuous development in an EU research program) is suitable for the data collection of various risk factors. It can be used to determine whether the hearing loss is occupational in origin and to estimate the efficiency of hearing conservation measures. NoiseScan also predicts the development of hearing loss in individuals in 5-year periods. The goal is to improve and validate the rules by which single and combined risk factors contribute to HIHL, thus leading to more precise prediction of individual hearing loss, and for the evaluation of success of the hearing conservation programs.
Asunto(s)
Bases de Datos como Asunto , Pérdida Auditiva Provocada por Ruido/prevención & control , Ruido/efectos adversos , Enfermedades Profesionales/prevención & control , Audiometría/métodos , Dispositivos de Protección de los Oídos/normas , Audición/fisiología , Pérdida Auditiva Provocada por Ruido/diagnóstico , Humanos , Exposición Profesional/efectos adversosRESUMEN
The peak sound pressure level (SPL), spreading of pressure wave and other physical characteristics of the impulse noise from weapons were studied in actual shooting conditions for assessment of gunfire noise exposure. Additionally, the attenuation of SPL by hearing protectors was measured with miniature microphones to evaluate protection efficiency in real shooting conditions. The peak SPLs at the shooter's ear ranged from 132 dB (miniature rifle) to 183 dB (howitzer). The spectral content of the main part of the acoustic energy was less than 400 Hz (peak 16-100 Hz) for large-caliber weapons and 150-2,500 Hz (peak 900-1,500 Hz) for small-caliber weapons (rifles). The safe distances from the noise source (less than 140 dB peak SPL) were 50-200 m for large-caliber weapons. Rifle impulses (assault rifle, caliber 7.62) had a peak SPL of 154 dB at a distance of 4 m from the muzzle. The peak SPLs of different explosives ranged from 125 to 185 dB at distances of 10 to 300 m. In rifle shooting, the attenuation efficiency of earplugs (16dB) or small-volume (thin) earmuffs (17 dB) was not sufficient and their use as sole protectors cannot be recommended. Instead, large-volume earmuffs should be used. Impulses from pistol and shotgun were fairly effectively attenuated both by small-volume and large-volume earmuffs. All kinds of earmuffs appeared to be ineffective (attenuation less than 15 dB) against impulses from large-caliber weapons with energy content at very low frequencies. Therefore, the combined use of earmuffs and earplugs is recommended for the most noisy operations. On the basis of the present data, wider safety zones were adopted in the Finish Defence Forces at shooting with different weapons.
Asunto(s)
Armas de Fuego , Ruido/efectos adversos , Dispositivos de Protección de los Oídos/estadística & datos numéricos , HumanosRESUMEN
The earmuff attenuation of acoustic impulses produced by large-caliber weapons was measured with a high-speed microcomputer controlled unit. The estimated accuracy was +/- 1 dB in peak sound-pressure level measurements. The peak levels outside earmuffs were 184 dB for the heavy bazooka and 172 dB for the hand-held bazooka (re: 20 microPa). Heavy bazooka impulse peak levels were attenuated from 7 to 19 dB by the earmuffs depending on the mass and volume of the measured three types of earmuffs. Hand-held bazooka impulse peak levels were attenuated by the earmuffs from 9 to 15 dB. The risk limits for hearing loss from a single impulse were exceeded in spite of the use of earmuffs when the criteria of CHABA (USA) or Pfander (Germany) were applied. The unexpectedly low attenuation was due to the low-frequency waveform of the high-level impulses. The earmuffs were found to prolong the impulse duration, which may reduce the benefit otherwise achieved by attenuation of the peak levels.
Asunto(s)
Acústica , Dispositivos de Protección de los Oídos , Armas de Fuego , Pérdida Auditiva Provocada por Ruido/epidemiología , Pérdida Auditiva Provocada por Ruido/prevención & control , Humanos , Ruido , Presión , Factores de RiesgoRESUMEN
The peripheral circulation was studied in 19 lumberjacks and in 12 control subjects. Twelve of the lumberjacks were free from vascular symptoms and seven had vibration induced white finger (VWF). Using the strain-gauge plethysmographic technique, the digital circulation was examined at rest, during cooling of the upper body, and during heating of the upper body. At rest and during vasodilatation no significant differences were found between the lumberjacks and the controls. During reflexive vasoconstriction, digital blood flow in the upper body was more reduced in lumberjacks with VWF than in control subjects. Furthermore, digital blood pressure of the lumberjacks with VWF fell more than in the control group. The peripheral resistance also increased more, but this difference was not statistically significant. There was no evidence that the exaggerated vasoconstriction of VWF resulted from a narrowing of the lumen of arterioles due to hypertrophy of the vessel wall. The present findings suggest that VWF is produced by the highly sensitive responsiveness of the affected vessel to normal vasoconstrictor stimuli.