An 8-week brain MRI follow-up analysis of rat eosinophilic meningitis caused by Angiostrongylus cantonensis infection.

Early differential diagnosis and timely follow-up are advantageous in the management of Angiostrongylus cantonensis infection. This study aimed to characterize angiostrongyliasis in the rat brain for an 8-week period using magnetic resonance imaging (MRI) with contrast-enhanced T1-weighted images (T1WI), T2-weighted imaging (T2WI), fluid attenuation inversion recovery (FLAIR) and R2 mapping sequences. The data were analysed with Mathematica and Matlab software programs for weekly changes in each brain following the infection of 20, 50, 100 and 300 third-stage larvae (L3), respectively. The results showed that the average subarachnoid space detected by T2WI technique was peaked up to 10% increase of original size on day 35 after 100 or 300 larvae infection, while those infected with 20 or 50 larvae showed less than 4% increase during the entire course of observation. This increase was relevant to the mortality of the infected rats, because those with 100 or 300 larvae infections showed a sharp decrease in survival rate before day 40. After day 40, the average subarachnoid space was decreased, but the average ventricle size was persistently increased, with the highest increase observed in the group infected with 300 larvae on day 56. Furthermore, the R2 mapping mean and R2 mapping size were significantly different between the brains with severe infection (100 and 300 larvae groups together) and those with mild infection (20 and 50 larvae groups together) on day 49, but not on day 35. Our results showed that diagnosis for different quantity of larvae infection using MRI is possible and follow-up characterization is informative in revealing the effects of angiostrongyliasis on different brain areas. In conclusion, our results support the use of MRI as a non-invasive diagnostic technique for eosinophilic meningitis caused by A. cantonensis infection.

Efficacy of albendazole-GM6001 co-therapy against Angiostrongylus cantonensis-induced meningitis in BALB/c mice.

Angiostrongylus cantonensis causes a form of parasitic meningitis in humans. Albendazole kills the nematode larvae staying in the brain. However, the dead larvae are capable of evoking a severe inflammatory response resulting in the brain damage. Matrix metalloproteinase 9 (MMP-9) is associated with the development of meningitis and with the immune inflammatory reaction. Presently, we studied the combination effects of albendazole and GM6001 (a MMP-9 inhibitor) against angiostrongyliasis in BALB/c mice. Co-administration of drugs produced marked effects; to kill the infecting larvae and to block MMP-9 activity. The combination treatment reduced MMP-9 activity by 89.2% in cerebrospinal fluid. The numbers of inflammatory cells increased significantly upon establishment of infection, but subsided upon co-treatment. Significantly fewer larvae were recovered from treated mice than from untreated, infected mice. The present results strongly suggest that co-therapy with albendazole and GM6001 may be an useful approach for the treatment of human angiostrongyliasis.

Induction of matrix metalloproteinase-9 in murine eosinophilic meningitis caused by Angiostrongylus cantonensis.

Matrix metalloproteinases (MMP) have been implicated in the pathogenesis of various inflammatory diseases of the central nervous system. In the present study, a gelatinase was found to be induced in parasitic meningitis caused, in mice, by Angiostrongylus cantonensis. The enzyme had a molecular weight of about 94 kDa, showed maximal activity between pH 6 and pH 8, and was clearly inhibited by EDTA and 1,10-phenanthroline but not by leupeptin or phenylmethanesulphonyl fluoride. When samples of cerebrospinal fluid from the mice with meningitis were blotted with specific antiserum against gelatinase B (MMP-9), a 94-kDa immunopositive band was observed, indicating that the induced gelatinase was MMP-9. In the A. cantonensis-infected mice, immuno-histochemistry demonstrated MMP-9 within the endothelial cells lining the vascular spaces of the brain and in the leucocytes that were found, in aggregates, in the subarachnoid space. Leucocytes may play an important role in the pathogenesis of inflammatory disorders of the central nervous system.

The detection of impedance cardiogram characteristic points using wavelet transform.

A novel impedance cardiograph event detection method using wavelet transform is proposed. When compared to the C and E points in the pressure-volume loop, the wavelet method performs significantly better than the traditional method (P < 0.05) in the B and X points detection even after the addition of 20% artificial noise into the test signal. Nevertheless, the SVs estimated by ICG are poorly correlated with values measured by the conductance catheter.

The effect of data reduction by independent component analysis and principal component analysis in hand motion identification.

Both independent component analysis (ICA) and principal component analysis (PCA) were used in this study to evaluate their effects in data reduction in the hand motion identification using surface electromyogram (SEMG) and stationary wavelet transformation. The results indicate that both methods increase the number of training epochs of the artificial neural network. The unsupervised fast ICA reduces the number of SEMG channels from 7 to 4. However the hand motion identification rate using the reduced channels is significantly lower (p < 0.05). On the other hand, the PCA reduces the size of neural network by more than 70%. Moreover, the results of discrimination rate and neural network training epochs show no significant difference as compared to the results before PCA reduction. The result of this study demonstrates that using wavelet and PCA are effective pre-processing for surface EMG analysis. It can efficiently reduce the size of neural network and increase the discrimination rate for different hand motions.

Prolonged fractionation of paced right atrial electrograms in patients with atrial flutter and fibrillation.

OBJECTIVES: This study investigated the extent of fractionation of paced right atrial electrograms in patients with and without paroxysmal atrial flutter (AFL) or atrial fibrillation (AF). BACKGROUND: Slow conduction through nonuniform anisotropic atrial muscles, represented by fractionated electrograms, may favor the generation of atrial tachyarrhythmias. METHODS: This study included 10 control patients (Group 1), 8 patients with documented paroxysmal AFL (Group 2) and 10 patients with documented paroxysmal AF (Group 3). Five electrode catheters were placed in the different sites of the right atrium and one catheter was positioned at the coronary sinus ostium. Atrial pacing from one site was done by a constant drive train with an extrastimulus inserted every fourth beat while recording at the other five sites was performed. The delay of each fractionated potential in the high-pass filtered atrial electrogram in response to extrastimulation was determined and used to construct conduction curves of delay versus the S1S2 interval. RESULTS: The mean increase in electrogram duration between a coupling interval of 350 ms and 10 ms above atrial refractoriness was significantly greater in Groups 2 and 3 compared with that in Group 1 (8.5 +/- 2.5 vs. 11.0 +/- 2.7 vs. 5.9 +/- 2.3 ms, respectively, p < 0.001). The mean S1S2 interval at which delay increased suddenly was also longer in Groups 2 and 3 compared with Group 1 (326 +/- 9 vs. 343 +/- 12 vs. 307 +/- 17 ms, respectively, p < 0.001). CONCLUSIONS: Increased delays in the individual potential of the fractionated atrial electrograms may be related to the development of AFL and AF.

Development of brain injury in mice by Angiostrongylus cantonensis infection is associated with the induction of transcription factor NF-kappaB, nuclear protooncogenes, and protein tyrosine phosphorylation.

Eosinophilic meningitis or meningoencephalitis caused by Angiostrongylus cantonensis is endemic to the Pacific area of Asia, especially Taiwan, Thailand, and Japan. Although eosinophilia is an important clinical manifestation of A. cantonensis infection, the role of eosinophils in the progress of the infection remains to be elucidated. In this experiment, we showed that A. cantonensis-caused eosinoplia and inflammation might lead to the induction of NF-kappaB and protooncogene expression via activation of the tyrosine phosphorylation signal pathway. After mice were infected daily with 30 third-stage larvae of A. cantonensis by oral adminstration for 6 weeks, no significant differences PKC-alpha, MEK-1, ERK-2, JNK, and p38 protein expression were found between the control and infected mice. However, the protein tyrosine phosphorylation levels, NF-kappaB, and iNOS protein products were significantly increased by 3.5-, 3.3-, and 6.3-fold, respectively, after 3 weeks of A. cantonensis infection. The same pattern was found for c-Myc, c-Jun, and c-Fos proteins, which were elevated by 3.2-, 2.3-, and 3.4-fold, respectively, compared to control animals after 3 weeks. The expression potency of these proteins started increasing in week 1, reaching maximal induction in week 3, and then declining in week 5 after A. cantonensis infection. Another consistent result was noted in the pathological observations, including eosinophilia, leukocyte infiltration, granulomatous reactions, and time responses in brain tissues of infected mice. These data suggest that the development of brain injury by eosinophlia of A. cantonensis infection is associated with NF-kappaB and/or nuclear protooncogenes expression, which is activated by the tyrosine phosphorylation pathway.

In vitro effects of various neuropharmacological agents on the motility of adult Clonorchis sinensis.

Stimulatory effects were observed in vitro when 5-HT was applied to whole worm preparations of Clonorchis sinensis. The stimulatory action of 5-HT was antagonized by tryptophol, clomipramine and imipramine. Strychnine remarkably stimulated motility. The action of strychnine was enhanced by 5-HT but inhibited by tryptophol. GABA, Phenylephrine, isoproterenol, dibenamine and phentolamine showed little influence on motility. Inhibitory effects were caused by eserine, arecoline, carbachol, DMPP and hemicholinium-3, but not by ACh, pilocarpine and McN-A-343. The inhibitory actions of eserine or carbachol were not antagonized by d-tubocurarine, hexamethonium or atropine. These results suggest that in regulating the motility of C. sinensis ACh may function as an inhibitory neurotransmitter and 5-HT as an excitatory transmitter in a manner similar to that already established for other trematodes such as Fasciola hepatica and Schistosoma mansoni.

A methodological improvement and system validation to obtain precise behavioral parameters for schedule-induced polydipsia.

A Schedule-Induced Polydipsia (SIP) animal behaviors monitor system was devised. The system included a software package to acquire data and to format the data storage of the animal behavior in the experiment was designed. Data analysis software will also extract the necessary information from recorded data. The SIP experimental apparatus consists of a number of subsystems. They are operant chambers, event counters, signal acquisition and storage system. The operant chamber is equipped with hopper, pellet and lick sensor and locomotion detector. The data acquisition and storage subsystem are custom designed that run on IBM-PC. The discrete time markers for pellet drop, bar pressing and licking water were recorded at 200 Hz. The image of rat locomotion was recorded at 10 Hz. A test of four different stages of animal to verify the accuracy of the system was reported in this article. The food-deprived rats that were exposed to the intermittent food schedule (1 per minute) exhibit an excess drinking behavior. The animal behaviors that were monitored during the scheduled events are adjunctive, facultative and terminal behavior. The volume of water intake was also recorded for reference. The real time data is stored chronologically into two types of data file. Therefore, a special data analysis procedure is designed to extract the results for off-line statistic analysis. The extracted parameters for animal behaviors analyzing including drinking efficiency, inter-lick intervals, number of drinking bursts, size of burst, temporal distribution of licking, temporal distribution of bar pressing and locomotion. The system can be employed in many different SIP studies such as investigating the effect of acute and chronic influence of amphetamine.

Construction of a screening inventory for major mental disorder in the Army.

The development of a 29-item self-report inventory to screen for non-adaptiveness with an episode or a history of major mental disorder in the army is described. Initially, a 58-item inventory was constructed. However, through a pilot study 46 items remained. The reliability and validity test of the 46-item inventory was then carried out on 357 patients and 126 controls. Firstly, item analyses were performed which showed 12 items that distinguished between schizophrenic patients and controls, 9 items that distinguished between depressive patients and controls and 8 items that distinguished between manic patients and controls. A final 29-item inventory was found to have satisfactory reliability, sensitivity and specificity. The analyses of the best cut-off scores using the receiver-operating-characteristic curve suggested a score of 3 for the Schizophrenia Scale, 2 for the Depression Scale, and 3 for the Mania Scale. The inventory can be completed in about 5 to 10 minutes and has a simple method of scoring. The use of this screening inventory in the army is discussed.

L ferritin accumulation in macrophages infiltrating the lung during rat Angiostrongylus cantonensis infection.

A 22-kDa protein was increased quantitatively, as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis techniques, in lung microsomes prepared from Angiostrongylus cantonensis-infected rats. However, it was almost absent in normal rats. The protein was purified by sequential chromatography on Superdex 200 columns and identified chemically and immunologically as ferritin. Using isoelectric focusing and anion exchange chromatography, it was identified as L ferritin. Distribution of this 22-kDa protein in the lung tissue of A. cantonensis-infected rate was studied by immunocytochemistry. Positively stained cells were mainly infiltrated macrophages. Our results suggest that L ferritin accumulation in the macrophages may be related to the proliferation of connective tissue elements and the inflammatory response to A. cantonensis dwelling in the pulmonary arteries of the rat.

In vitro effects of VD-99-11 on Angiostrongylus cantonensis and isolated frog rectus.

In vitro effects of VD-99-11 were examined using adult Angiostrongylus cantonensis and isolated frog rectus. In A. cantonensis, paralysis was elicited by VD-99-11 at 10(-9)-10(-6) g/ml. The paralysis caused by VD-99-11 (10(-8) g/ml) was antagonized by picrotoxin or bicuculline but not by phentolamine. A relationship between VD-99-11 and gabergic antagonists was observed in worm preparations contracted by eserine or pyrantel: VD-99-11 at higher concentrations (3x10(-6) g/ml) caused a marked contraction. In worm preparations contracted with eserine or pyrantel, the only additional contraction induced by VD-99-11 (5x10(-6) g/ml) was antagonized by strychnine. In experiments on the guanidine (2.5x10(-3) M)-induced twitch responses in isolated frog rectus, marked stimulation was caused by VD-99-11 (3-5x10(-6) g/ml). The stimulated responses induced by VD-99-11 were antagonized by tetrodotoxin, D-tubocurarine, strychnine, and hemicholinium-3, respectively. These results suggest that VD-99-11 seems superior to milbemycin D, milbemycin oxime, and ivermectin in some aspects, such as in vitro potency, though this new substance is similar to these drugs in having two different actions on the gabergic mechanism at lower concentrations and on the cholinergic mechanism at higher concentrations.

Signal fidelity requirements for deriving impedance cardiographic measures of cardiac function over a broad heart rate range.

Our findings indicate that the impedance cardiogram spectrum extends from DC to 50 Hz. Any amplifier with an upper band limit less than 50 Hz can be expected to produce attenuation and distortion of the impedance cardiogram. This signal attenuation may be systematically enhanced under conditions of high heart rate when a greater proportion of signal energy will be in the upper frequency range of the impedance cardiogram spectrum. Therefore, the present study was designed to assess the influence of amplifier bandwidth on dZ/dtmax, stroke volume, and systolic time intervals (LVET, PEP, QZ, QX). Simultaneously measured delta Z and dZ/dt signals from two impedance cardiographs, with corner frequencies of 120 and 60 Hz for the delta Z and 50 and 15 Hz for dZ/dt channels, were contrasted over a broad range of heart rate (70-150 bpm). In addition to the analog dZ/dt signals obtained from the instruments, the delta Z signals were digitally converted to dZ/dt by off-line digital differentiation with a 50 Hz corner frequency. The results demonstrated that the measurements with the 15 Hz corner frequency, when compared with the 50 Hz corner frequency measurements, systematically attenuated the dZ/dtmax amplitude and stroke volume measurements as heart rate increased. The attenuation of dZ/dtmax and stroke volume ranged from about 13% to 26% as heart rate increased from 70 to 150 bpm. When the upper bandlimit was 50 Hz, the dZ/dt signal had greater resolution of waveform events and produced less prolonged systolic time intervals. The 15 Hz amplifier differentially influenced the B point, Z-peak and X minimum, having no apparent effect on the temporal location of the B point, but delaying the Z-peak about 21.7 ms and the X minimum about 7.4 ms. These findings indicate that impedance cardiographs with insufficient upper bandlimits will differentially influence ICG-derived measurements as heart rate varies.

[Experimental control study of fasciolopsiasis in Taiwan].

This study was conducted from March 1985 to September 1987 among the pupils of Chung-Ying Primary School of Hsia-Ying township in Tainan Country, Southern Taiwan. Health education revealed the preferred control method against Fasciolopsis infection in this study. The education method included lectures, demonstrations of parasites and home visits at regular intervals. The subjects included the staff and children of the school and all family members. At the same time, the children infected with Fasciolopsis were treated with praziquantel in June, 1986. Fecal material of the all primary school children of Hsia-Ying township was examined with the antiformin-ether concentration method. One year before control the positive rate of Fasciolopsis was 25.1%. One year after control in March 1986, it was decreased to 9.5%. Two and a half years after control in September 1987, it was farther brought down to 1.0%. The x2-test of positive rate showed statistically different. Health education with medication is recommended for the control of fasciolopsiasis.

Microcomputer servo-controlled bicycle ergometer system for psychophysiological research.

Steady state exercise is widely used for psychophysiological studies in which a constant heart rate at a predetermined level is desired. We have developed a microcomputer servo-controlled bicycle ergometer system that can be used for administering steady state exercise. Fourteen healthy male subjects, with a wide range of fitness levels (measured by VO2max) were exercised to either a fixed workload (130 watts) or a predetermined heart rate level (servo-heart rate) of 122 bpm (i.e., 65% of maximum calculated heart rate for the sample). Servo-heart rate was implemented using a feedback loop that automatically adjusted workload to compensate for immediate variations in heart rate, resulting in a more consistent heart rate. Heart rate varied from the predetermined value by 17 bpm during fixed workload but only 3 bpm during servo-heart rate (p less than .05). Therefore, by using the microcomputer servo-controlled bicycle ergometer, heart rate was maintained at a predetermined level regardless of the subject's fitness level. VO2max and workload during servo-heart rate were significantly correlated (r = .85, p less than .05). Therefore, the workload necessary to maintain heart rate at a constant level may provide an approximate index of aerobic fitness level.

New signal processing techniques for improved precision of noninvasive impedance cardiography.

Impedance cardiographic determination of clinically important cardiac parameters such as systolic time intervals, stroke volume, and related cardiovascular parameters has not yet found adequate application in clinical practice, since its theoretical basis remains controversial, and the precision of beat-to-beat parameter estimation has until recently suffered under severe shortcomings of available signal processing techniques. High levels of noise and motion artifacts deteriorate signal quality and result in poor event detection. To improve the precision of impedance cardiography, new techniques for event detection and parameter estimation have been developed. Specifically, matched filtering and various signal segmentation and decomposition techniques have been tested on impedance signals with various levels of artificially superimposed noise and on actual recordings from subjects in a laboratory study of cardiovascular response to a cognitive challenge. Substantial improvement in the precision of impedance cardiography was obtained using the newly developed signal processing techniques. In addition, some preliminary evidence from comparisons of the impedance cardiogram with invasive aortic electromagnetic flow measurement in anesthetized rabbits is presented to address questions relating to the origin of the impedance signal.

[A preliminary study on epidemiology of fasciolopsiasis in Tainan Hsien, south Taiwan].

Stool samples of 817 children (grades 4-6) from 3 primary schools, were collected and examined by the antiformin-ether concentration technique. Results showed no cases of Fasciolopsis were found from Liu-Ying primary school (Liu-Ying Hsiang), but infection rates of this fluke were found to be 13.0% and 3.5% for Ho-Chien and Tung-Hsing primary schools (Hsia-Ying Hsiang) respectively. Fourteen infected children had recent experience of eating raw water caltrops. On the other hand, only 8(21.6%) of 37 non-infected children had eaten raw water caltrops. Examination of intermediate host (Segmentina hemisphaerula, Trapa bispinosa) results showed that neither cercaria nor metacercaria was found. Intestines and stool samples from pigs were examined for worms and eggs of Fasciolopsis, the results were also negative.