Acetylcholine and Choline in Neuronal Tissue Measured by HPLC with Electrochemical Detection

Abstract: A simple, rapid method is presented for the determination of acetylcholine (ACh) and choline (Ch) in neuronal tissue using HPLC with electrochemical detection. The method is based on the separation of ACh and Ch by reverse-phase HPLC and mixing the effluent as it emerges from the column with acetylcholinesterase and Ch oxidase, which converts endogenous Ch and Ch produced by the hydrolysis of ACh to betaine and hydrogen peroxide. Production of hydrogen peroxide is continuously monitored electrochemically. The sensitivity of the procedure is 1 pmol for Ch and 2 pmol for ACh. Specificity of the method is based on HPLC, two specific enzymatic reactions, and the detection of hydrogen peroxide.     

Principles and applications of luminescence spectrometry coupled to liquid chromatographic techniques

An overview is presented of the physicochemical basis of luminescence, and its application to the detection of chemicals (drugs, biomedically important compounds, environmentally active substances) in liquid chromatographic systems.     

Integration time for short broad band clicks in echolocating FM-bats (Eptesicus fuscus)

Vespertilionid FM-bats (four Eptesicus fuscus and one Vespertilio murinus) were trained in an electronic phantom target simulator to detect synthetic echoes consisting of either one or two clicks. The threshold sound pressure for single clicks was around 47 dB peSPL for all five bats corresponding to a threshold energy of -95 dB re 1 Pa² * s. By varying the interclick interval, T, for double clicks it was shown that the threshold intensity was around — 3 dB relative to the threshold for single clicks at T up to 2.4 ms, indicating perfect power summation of both clicks. A threshold shift of -13.5 dB for a 1 ms train of 20 clicks (0.05 ms interclick interval) confirmed that the bats integrated the power of the stimuli. At T longer than around 2.5 ms the threshold for double clicks was the same as for single clicks. Thus, the bats performed like perfect energy detectors with an integration time of approximately 2.4 ms. This integration time is an order of magnitude shorter than that reported for bats listening passively for pure tones. In our setup the bats emitted sonar signals with durations of 2–3 ms. Hence, the results may indicate that while echolocating the bats integration time is adapted to the duration of the sonar emissions.Abbreviations AGC automatic gain control - FM frequency modulated - peSPL peak equivalent sound pressure level - rms root mean square - SD standard deviation - SE standard error of mean - T interclick interval     

PCR detection of MLOs in quick decline-affected pear trees in Italy

Polymerase chain reaction (PCR) amplification, using primers derived from the 16S rRNA gene, followed by restriction fragment length polymorphism (RFLP) analysis with Alu I restriction endonuclease was used to detect myc-oplasma-like organisms (MLOs) associated with pear decline. MLOs were consistently detected in pear trees that suddenly wilted and died within a few days during summer, as well as in pears of the same orchards with symptoms similar to the slow form of pear decline. In both cases the same RFLP pattern was obtained. Declining pear trees were 5 to 8-yr-old cvs Williams, Kaiser and Max Red Bartlett grafted on to Pyrus communis seedling rootstocks. All the orchards affected by quick decline had severe attacks of pear psyllid (Cacopsylla pyri) during the year this study was performed and during the previous year. The results showed the suitability of DNA amplification by the polymerase chain reaction for the detection of pear decline MLOs and established that MLOs can be detected in infected tissues of dead trees.     

Efficiency of serum copper/zinc ratio for differential diagnosis of patients with and without lung cancer

We examined serum copper (Cu), serum zinc (Zn), and the serum copper/zinc ratio (Cu/Zn) in 162 patients. All of them were seen to have an abnormal shadow in the chest X-ray films, that is, 109 patients with lung cancer (LC) and 53 patients with no lung cancer (NLC). The mean Cu and Cu/Zn in LC patients were significantly higher than those in NLC patients (p<0.05). In LC patients, Cu and Cu/Zn were higher and Zn was lower in advanced tumors than early ones. There was a significantly clear relation between Cu or Cu/Zn and the tumor (T) stages. When the relative risk (RR) of LC was estimated, it was seen that the higher Cu and Cu/Zn became, the higher RR became. Furthermore, we showed the sensitivity of the receiver operator characteristic of the test (ROC) curve for Cu, Cu/Zn, and carcinoembryonic antigen (CEA) to diagnose LC, as explained in a paragraph of methods.The determinations of Cu, Zn, and Cu/Zn are simple and inexpensive. They also appear to have a great diagnostic value in determining the local invasion of LC and as a screening test in the high-risk patients for LC.     

Detection of gaps in sinusoids by frog auditory nerve fibers: importance in AM coding

Physiological studies were carried out in the frog (Rana pipiens pipiens) eighth nerve to determine: (i) whether the modulation rate or the silent gap was the salient feature that set the upper limit of time-locking to pulsed amplitude-modulated (PAM) stimuli, (ii) the gap detection capacity of individual eighth nerve fibers. Time-locked responses of 79 eighth nerve fibers to PAM stimuli (at the fiber's characteristic frequency) showed that the synchronization coefficient was a low-pass function of the modulation rate. In response to PAM stimuli having different pulse durations, a fiber gave rise to non-overlapping modulation transfer functions. The upper cut-off frequency of time locking was higher when tonepulses in PAM stimuli had shorter duration. The fact that the cut-off frequency was different for the different PAM series suggested that the AM rate was neither the sole, nor the main, determinant for the decay in time-locking at high AM rates. Gap detection capacity was determined for 69 eighth nerve fibers by assessing fiber's spiking activities to paired tone-pulses during an OFF-window and an ON-window. It was found that the minimum detectable gap of eighth nerve fibers ranged from 0.5 to 10 ms with an average of 1.23–2.16 ms depending on the duration of paired tone pulses. For each fiber, the minimum detectable gap was longer when the duration of tone pulses comprising the twin-pulse stimuli was more than four times longer. When the synchronization coefficient was plotted against the silent gap between tones pulses in the PAM stimuli, the gap response functions of a fiber as derived from multiple PAM series were equivalent to gap response functions deriving from twin-pulse series suggesting that it was the silent gap which primarily determined the upper limit of time-locking to PAM stimuli.Abbreviations MTF modulation transfer function - PAM pulse amplitude modulated - SAM sinusoidally amplitude modulated - SC synchronization coefficient - TW time window     

Bayesian signal extraction from noisy FT NMR spectra

Summary The statistical interpretation of the histogram representation of NMR spectra is described, leading to an estimation of the probability density function of the noise. The white-noise and Gaussian hypotheses are discussed, and a new estimator of the noise standard deviation is derived from the histogram strategy. The Bayesian approach to NMR signal detection is presented. This approach homogeneously combines prior knowledge, obtained from the histogram strategy, together with the posterior information resulting from the test of presence of a set of reference shapes in the neighbourhood of each data point. This scheme leads to a new strategy in the local detection of NMR signals in 2D and 3D spectra, which is illustrated by a complete peak-picking algorithm.     

Effect of copper sulfate on experimental atherosclerosis

Serum copper concentration increases significantly (p<0.01) in rats with experimental atherosclerosis compared to a control group. The serum zinc, the zinc, and copper concentration in abdominal aorta and in liver decreases significantly (p<0.05) compared to the control group. Administration of copper sulfate for 100 d in these animals induces a significant increase of serum copper (p<0.01), decrease of serum cholesterol (p<0.05) and increase of liver copper concentration as compared with the group fed only a high cholesterol diet. In the aorta of these animals the copper concentration increases and edema and lipid infiltration are considerably less than in the group of animals fed only a high lipid diet.     

Long-term changes in frequencies ofPythium iwayamai andP. paddicum in upland- and paddy-field soils

The long-term changes in the frequencies ofPythium iwayamai andP. paddicum in upland- and flooded paddy-field soils were compared. The frequencies of both species fell in both soils within the first 6 months after they had been buried. After 36 and 48 months,P. iwayamai showed higher frequency thanP. Paddicum in upland-field soil, whileP. paddicum showed higher frequency thanP. iwayamai in paddy-field soil. These findings indicate respectively higher colonization ability to organic matters ofP. iwayamai in upland-field soil andP. paddicum in paddy-field soil.This study was partly supported by a Grant-in-Aid (no. 63560048) from the Ministry of Education, Science and Culture, Japan.