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Pattern analysis of sleep-deprived human EEG.

Progress during the past decade in non-linear dynamics and instability theory has provided useful tools for understanding spatio-temporal pattern formation. Procedures which apply principle component analysis (using the Karhunen-Loeve decomposition technique) to the multichannel electroencephalograph (EEG) time series have been developed. This technique shows localized changes of cortical functioning; it identifies increases and decreases of the activity of localized cortical regions over time while the subject performs a simple task or test. It can be used to demonstrate the change in cortical dynamics in response to a continuous challenge. Using 16 EEG electrodes, the technique provides spatio-temporal information not obtained with power spectrum analysis, and includes the weighted information given with omega complexity. As an application, we performed a pattern analysis of sleep-deprived human EEG data in 20 healthy young men. Electroencephalograph recordings were performed on subjects for <2 min, with eyes closed after normal sleep and after 24 h of experimentally-induced sleep deprivation. The significant changes in the eigenvector components indicated the relative changes of local activity in the brain with progressive sleep deprivation. A sleep deprivation effect was observed, which was hemispherically correlated but with opposite directional dynamics. These changes were seen in the temporo-parietal regions bilaterally. The application of the technique showed that the simple test task was performed with a limited unilateral hemispheric involvement at baseline, but needed a much larger cortical participation with decreased frontal activity and increased coherence and bilateral hemispheric involvement. The calculations performed demonstrated that the same weighted changes as those obtained with omega complexity were shown, but the technique had the added advantage of showing the localized directional changes of the principle eigenvector at each studied electrode, pointing out the cortical localized region affected by the sleep deprivation and toward which direction the environmental challenge induced the spatial change. This methodology may allow the evaluation of changes in local dynamics in brain activity in normal and pathological conditions.

Adult↗

Relative influence of age, resting heart rate and sedentary life style in short-term analysis of heart rate variability.

In order to assess the relative influence of age, resting heart rate (HR) and sedentary life style, heart rate variability (HRV) was studied in two different groups. The young group (YG) consisted of 9 sedentary subjects aged 15 to 20 years (YG-S) and of 9 nonsedentary volunteers (YG-NS) also aged 15 to 20. The elderly sedentary group (ESG) consisted of 16 sedentary subjects aged 39 to 82 years. HRV was assessed using a short-term procedure (5 min). R-R variability was calculated in the time-domain by means of the root mean square successive differences. Frequency-domain HRV was evaluated by power spectrum analysis considering high frequency and low frequency bands. In the YG the effort tolerance was ranked in a bicycle stress test. HR was similar for both groups while ESG showed a reduced HRV compared with YG. Within each group, HRV displayed a negative correlation with HR. Although YG-NS had better effort tolerance than YG-S, their HR and HRV were not significantly different. We conclude that HRV is reduced with increasing HR or age, regardless of life style. The results obtained in our short-term study agree with others of longer duration by showing that age and HR are the main determinants of HRV. Our results do not support the idea that changes in HRV are related to regular physical activity.

Adolescent↗

[A chromatographic analysis of the Mycobacterium tuberculosis strains isolated from an outbreak in HIV patients in Cuba].

A group of strains of Mycobacterium tuberculosis isolated from an outbreak in HIV-infected patients was studied by chromatographic techniques. A group of strains of M. Tuberculosis from symptomatic respiratory patients (SR+ 14) and patterns strains from the laboratory collection were used as a reference aimed at making a qualitative comparison of the chromatographic patterns described by the strains isolated from patients. The chromatographic profiles of the strains isolated from patients (SR+) and fro HIV+ were obtained and compared by thin layer chromatography (TLC). Each of the present fatty acids was identified by using the gas chromatography technique (GC) coupled to mass spectrum analysis. All the studied strains were classified as Mycobacterium tuberculosis. According to the results attained, the usefulness of the chromatographic techniques as alternative techniques for the mycobacterial diagnosis is demonstrated.

AIDS-Related Opportunistic Infections↗

Effects of high altitude acclimatization on heart rate variability in resting humans.

The sympatho-vagal nerve interaction at the heart was studied by means of power spectrum analysis of heart rate variability in seven Caucasians (aged 27-35 years) in resting supine and sitting positions before and during 35 days of a sojourn at 5050 m above sea level (asl) and in six Sherpas (aged 22-30 years) at high altitude only. A high frequency peak (HF)-central frequency between 0.20 and 0.33 Hz, a low frequency peak (LF)-central frequency between 0.08 and 0.14 Hz, and a very low frequency component (< 0.05 Hz), no peak observed, were found in the power spectrum in both positions and independent of altitude. The peak powers, as a percentage of the total power, were affected by both body position and altitude. At sea level the change from a supine to a sitting position yielded a decrease in percentage HF from 25 (SEM 1.9)% to 6.2 (SEM 1.5)% (P < 0.05) and a significant increase in the ratio between LF and HF powers (LF:HF) from 1.7 (SEM 0.4) to 6.9 (SEM 1.6). At altitude compared to sea level in the supine position, percentage HF decreased from 25% to 10.9 (SEM 1.0)% (P < 0.05) and the LF:HF ratio increased from 1.7 to 4.8 (SEM 0.7) (P < 0.05). No changes occurred at altitude in the sitting position either in the peak powers or in the LF:HF ratio, but the central frequency of HF peak increased significantly from 0.25 (SEM 0.02) Hz to 0.32 (SEM 0.01) Hz. In the Sherpas comparable results to the Caucasians were found in both body positions. The high LF:HF ratios observed at altitude in both body positions and groups would suggest that hypoxia caused a shift of sympatho-vagal nerve interaction at rest toward a dominance of the sympathetic system, which was found at sea level only in the sitting position. An acclimatization period of 10 days higher than 2850 m asl and 1 month at 5050 m asl did not modify the interactions of the autonomic systems.

Acclimatization↗

Detrimental sequelae on the hemodynamics of the upper left limb after subclavian flap angioplasty in infancy.

The long-term effect of two surgical techniques for repair of coarctation of the aorta in infancy, namely, resection and end-to-end anastomosis (RETE) and subclavian flap angioplasty (SFA) on the blood supply of the upper left limb, was quantified by Doppler spectrum analysis of blood flow velocities in the left brachial artery at rest and during postocclusive reactive hyperemia. Twenty-three patients participated in this study: nine patients after SFA (median age, 8 years), 14 patients after RETE (median age, 8 years), and 10 control subjects (median age, 9.5 years). At rest, a highly significant decrease of blood flow velocities in the left brachial artery was measured in all patients of the SFA group compared with those of the RETE and control groups, as documented by various Doppler spectrum parameters: maximal frequency of advancing curve (p = 0.0001), pulsatility index (p = 0.0005), and resistance index (p = 0.039). During reactive hyperemia, a moderate capacity of physiologic augmentation of blood flow velocities was observed in five patients of the SFA group. This capacity was marginal in two patients with complaints of claudication in the left upper limb during strenuous exercise, which can be related to the number of branches of the left subclavian artery ligated during operation. This study indicates that SFA in infancy may lead to compromised hemodynamics of the upper left limb with potential for symptoms of ischemia during exercise.

Anastomosis, Surgical↗

Analysis of the PilQ secretin from Neisseria meningitidis by transmission electron microscopy reveals a dodecameric quaternary structure.

PilQ is a member of the secretin family of outer membrane proteins and is specifically involved in secretion of type IV pili in Neisseria meningitidis, Neisseria gonorrhoeae, and Pseudomonas aeruginosa. The quaternary structure of PilQ from N. meningitidis was analyzed by transmission electron microscopy by using a negative stain. Single particle averaging was carried out with a total data set of 650 individual particles, which produced a projection map generated from 296 particles at an estimated resolution of 2.6 nm. Oligomeric PilQ adopts a donut-like structure with an external ring that is 16.5 nm in diameter surrounding a central cavity that is 6.5 nm in diameter. Self-rotation and power spectrum analysis demonstrated the presence of 12-fold rotational symmetry, showing that PilQ is organized as a ring of 12 identical subunits. A model of the type IV meningococcal pilus fiber, based on the X-ray crystal structure of the N. gonorrhoeae pilin subunit, fitted neatly into the cavity, demonstrating how PilQ could serve as a channel for the growing pilus fiber.

Bacterial Outer Membrane Proteins↗

[Degradation pathway of 17alpha-ethynylestradiol by Sphingobacterium sp. JCR5].

A bacterial strain, JCR5, which degrades 17alpha-ethynylestradiol (EE2), was isolated from activated sludge of wastewater treatment plant treating wastewater from pharmacy factory mainly producing contraceptive medicine in Beijing, China. Based on its morphology, physiology biochemical properties and 16S rDNA sequence analysis, this strain was identified as Sphingobacteriumn sp. JCR5. Strain JCR5 can grow on EE2 as sole carbon and energy source. The degradation process for EE2 with initial concentration of 30 mg x L(-1) was studied, and the result indicated that the degradation rate of EE2 within 10 days was 87%. Experiments of different substrates showed that strain JCR5 can grow on many substrates other than EE2 such as several steroidal estrogens (estrone, 17beta-estradiol, estriol and mestranol), the intermediates in contraceptive medicine processing and some aromatic compounds. Mass spectrum analysis demonstrated that EE2 was oxygenized to estrone (E1) firstly, and 2-hydroxy-2,4-dienevaleric acid and 2-hydroxy-2,4-diene-1, 6-dioic acid were the main catabolic intermediates during EE2 degradation. The former adopted a pathway that was analogical to the pathway of the previously reported testosterone-degrading Comamnonas testosteroni TA441, and the latter was a metabolite with a different cleavage position of 3-hydroxy-4,5-9, 10-disecoestrane-1(10), 2-diene-5,9, 17- trione-4-oic acid from the former.

Biodegradation, Environmental↗

Visual evoked spectrum array and interhemispheric variations.

An EEG spectrum analysis during visual stimulation was computed in 19 normal subjects. Visual stimulation consisted of trains of flashes at frequencies ranging between 2.5 and 20 flashes per second. Recordings were carried out simultaneously from the right and left occipital regions with bipolar and referential montages. Compressed spectral arrays were computed for eight-second epochs at each recording site using a fast Fourier transform. The ratio of the spectral energy from homologus regions of right and left hemispheres at each stimulation frequency was determined. The ratios were graphically displayed in a visual evoked spectrum array (VESA) ratio plot (VESA-GRAM); the mean of the ratio plot was designated the VESA coefficient. The range of variation for these measurements was determined for normal subjects. An application of the technique to patients with hemianopia showed abnormal VESA (characterized by smaller spectral amplitudes over the appropriate hemisphere), abnormal VESA ratio plots, and high VESA coefficients. These preliminary findings suggest that VESA may be a promising method to detect retrochiasmatic visual defects.

Adult↗

The nonlinear dynamical analysis of the EEG in schizophrenia with temporal and spatial embedding dimension.

We applied nonlinear dynamics theory to EEG analysis of schizophrenic patients and estimated the correlation dimension with both temporal embedding and spatial embedding. A higher D2 was found when using a time-delay embedding method. Especially at F7 and Fp1, a significant increase showed. We concluded that more complex activity occurred in certain lobes of schizophrenic patients. Using the spatial embedding method, a relative lower global correlation dimension was obtained. This shows that there might be a diffuse slow wave activity through a schizophrene's global cerebrum. Finally, we discuss the study from three angles of clinical semiology, spectrum analysis and neuropsychology and draw some conclusions about the relationship between the nonlinear analysis of schizophrenia EEG and clinical research. It seems that the theory of a nonlinear dynamics system is a powerful tool for EEG research and may prove useful in complementing visual analysis of EEG accompanied with other study means for brain electrical activity.

Adult↗

Detection, identification, and estimation of biological aerosols and vapors with a Fourier-transform infrared spectrometer.

Two experiments were conducted with a Fourier-transform infrared (FTIR) spectrometer. The purpose of the first experiment was to detect and identify Bacillus subtilis subsp. niger (BG) bioaerosol spores and kaolin dust in an open-air release for which the thermal contrast between the aerosol temperature and background brightness temperature is small. The second experiment estimated the concentration of a small amount of triethyl phosphate (TEP) vapor in a closed chamber in which an external blackbody radiation source was used and where the thermal contrast was large. The deduced BG (TEP) extinction spectrum (identification) showed an excellent match to the library BG (TEP) extinction spectrum. Analysis of the time sequence of the measurements coincided well with the presence (detection) of the BG during the measurements, and the estimated concentration of time-dependent TEP vapor was excellent. The data were analyzed with hyperspectral detection, identification, and estimation algorithms. The algorithms were based on radiative transfer theory and statistical signal-processing methods. A subspace orthogonal projection operator was used to statistically subtract the large thermal background contribution to the measurements, and a robust maximum-likelihood solution was used to deduce the target (aerosol or vapor cloud) spectrum and estimate its mass-column concentration. A Gaussian-mixture probability model for the deduced mass-column concentration was computed with an expectation-maximization algorithm to produce the detection threshold, the probability of detection, and the probability of false alarm. The results of this study are encouraging, as they suggest for the first time to the authors' knowledge the feasibility of detecting biological aerosols with passive FTIR sensors.

Journal Article↗

Identification of an antifungal lipopeptide from Bacillus amyloliquefaciens HAU3 inhibiting the growth of Fusarium graminearum using preparative chromatography and 2D-NMR.

UNLABELLED: The presence of fungal contamination and its mycotoxins in animal feed is pervasive, posing a significant threat to the well-being and performance of animals, as well as the safety of animal-derived food products. In this work, we screened a strain of Bacillus amyloliquefaciens (B. amyloliquefaciens) HAU3 that exhibits efficient antifungal activity against the growth of Fusarium graminearum (F. graminearum). The antifungal activity was detected in the supernatant, with 20% sterile supernatant demonstrating an impressive antifungal rate of 98.46% against F. graminearum. The antifungal activity of the strain was evaluated through spectrum analysis and silage trials, revealing its effective antifungal activity against multiple fungal species. Furthermore, the strain is capable of degrading ZEN and its derivatives. The targeted disruption of fungal mycelial membrane was observed using scanning electron microscopy and transmission electron microscopy. Additionally, staining with the reactive oxygen species (ROS)-sensitive fluorogenic dye DCFH-DA and propidium iodide (PI) revealed that the strain induces accumulation of ROS in fungal mycelia. The active compounds underwent further separation, purification, and detection. The prominent active peak was identified through mass spectrometry and magnetic resonance spectroscopy. The molecular structure of the active compounds was predicted to be lipopeptides composed of 8 amino acids known as fengycin. The whole genome sequencing and informatics analysis unveiled a total of 13 gene clusters responsible for the synthesis of secondary metabolites. The antifungal effects of B. amyloliquefaciens HAU3 are exerted through the synthesis of fengycin, which selectively targets and compromises the integrity of fungal mycelia membranes, thereby making it a potential biocontrol agent for mitigating mycotoxin contamination in feed. IMPORTANCE: Mycotoxin contamination in animal feed, predominantly driven by Fusarium graminearum, represents a persistent threat to livestock health and food chain integrity. Here, we report the isolation of a soil-derived Bacillus amyloliquefaciens HAU3, exhibiting potent and broad-spectrum antifungal activity alongside efficient biodegradation of zearalenone and its derivatives. Mechanistic dissection reveals that fengycin, the principal bioactive metabolite, compromises fungal membrane integrity and elicits intracellular oxidative stress, culminating in hyphal collapse. Genomic profiling uncovers a diverse repertoire of biosynthetic gene clusters underpinning secondary metabolite production. These findings establish strain HAU3 as a promising microbial chassis for the development of next-generation biocontrol strategies aimed at mitigating mycotoxin burden in agroecosystems.

Bacillus amyloliquefaciens↗

Effect of static stretch on fatigue of lumbar muscles induced by prolonged contraction.

Low back pain (LBP) is believed to be the result of fatigue of the back extensor muscles induced by prolonged contraction. Although static stretch has been considered to promote recovery from such muscle fatigue by relaxation, little is known about the effect of stretch on muscle fatigue, especially in the back extensor muscles. The purpose of this study was to investigate the effect of static stretch on prolonged contraction-inducedfatigue of the back extensor muscles using electromyographic (EMG) spectral analysis in ten healthy volunteers. Bipolar surface electrodes were placed on the longissimus and the multifidus muscles. EMG signals were collected during two trials of a Sorensen trunk-holding test for two minutes, with a five-minute rest period between the two trials (control test). All subjects were asked to perform the same trials, with one-minute of static stretch and a four-minute rest period (stretch test). Maintaining the knees toward the chest in supine position resulted in static stretch. The median frequency (MF) was calculated using a spectrum analysis program, and the MF slope over time was computed by linear regression analysis, and normalized with the y-intercept. The decreasing rate of the normalized MF slope between two trials in two tests was compared. The decreasing rate of the normalized MF slope of the second trial was larger than that of the first trial in the control test (p < 0.05), but no significant difference was observed in the stretch test. The results indicated that static stretch had a significant effect on the recovery from fatigue of the back extensor muscles, since it influenced the decreasing rate of the normalized MF slope.

Adult↗

Acoustic characteristics of rough voice: subharmonics.

This study investigates the relationship between rough voice and the presence of subharmonics, which correspond to smaller yet distinct peaks located between two consecutive harmonic peaks in the power spectrum. Spectrum analysis was undertaken in 389 pathologic voices, of which 20 had subharmonics. Although all 20 voices had roughness perceptually, 8 had normal jitter and/or shimmer. The degree of roughness had a significant inverse relationship with the frequency of subharmonics. By digital signal processing, sound samples with various types of subharmonics were synthesized and perceptually analyzed. Power and frequency of subharmonics in the synthesized sound also had significant relationships with the degree of roughness. Rough voice is acoustically characterized not only by jitter and shimmer but also by the presence of subharmonics in the power spectrum. Subharmonics are important acoustic properties for objective evaluation of rough voices.

Adult↗

[Compositional analysis of two metal crowns and its cements unearthed from a Ming Dynasty grave in China by SEM].

OBJECTIVE: To investigate the prosthodontic level of our country in Ming Dynasty (from A. D. 1368 to A. D. 1644) and provide our courtry's history of stomatological development with some details. METHODS: By means of range estimation and SEM, the structure and shape of the metal crowns on the two human teeth which were unearthed from a Ming Dynasty grave, numbered 0754/0046 and stored in Wujin museum of Jiangsu province, as well as the compositions of the crowns and its cements were examined and analysed preliminarily. RESULTS: The two teeth looked like upper anterior teeth and no marks of preparations were observed. The golden-yellow metal crowns might have been hammered and welded into shape. The spectrum analysis of SEM showed the crowns were made from gold-copper alloy and the cements contains Zinc. CONCLUSION: The defects of teeth could have been restored with extra-hard gold-copper alloy and cement luting materials during A. D. 1500-1600 in China.

China↗

An experimental proton magnetic resonance spectroscopy analysis on early stage of acute focal cerebral ischemia.

Using different models of focal cerebral ischemia, the temporal and spatial rules of metabolism and energy changes in the post-ischemia brain tissue were measured by proton magnetic resonance spectroscopy (1HMRS) to provide valuable information for judging the prognosis of acute focal cerebral ischemia and carrying out effective therapy. Nine healthy Sprague-Dawly rats (both sexes) were randomly divided into two groups: The rats in the group A (n = 4) were occluded with self-thrombus for 1 h; The rats in the group B (n = 5) were occluded with thread-emboli for 1 h. The 1H MRS at 30, 40, 50, 60 min respectively was examined and the metabolic changes of NAA, Cho and Lac in the regions of interest were semiquantitatively analyzed. The spectrum integral calculus area ratio of NAA, Cho, Lac to Pcr + Cr was set as the criterion. The values of NAA.Cho in the regions of interest were declined gradually within 1 h after ischemia, especially, the ratio of Cho/(Pcr + Cr), NAA/(Pcr + Cr) at 60 min had significant difference with that at 50 min (P < 0.05). The ratio of Lac/(Pcr + Cr) began to decrease at 40 min from initial increase of Lac in both A and B groups. MR proton spectrum analysis was a non-invasive, direct and comprehensive tool for the study of cellular metabolism and the status of the biochemical energy in acute ischemia stroke.

Animals↗

Monochromatism determined at a long-wavelength/middle-wavelength cone-antagonistic locus.

The foveal increment threshold spectral sensitivity function for a 500 msec raised cosine stimulus without spatial edges exhibits a sharp drop or "notch" in sensitivity that coincides with the wavelength of a long-wavelength adapting field. An appropriate name for this phenomenon is the "Sloan notch", after Louise Sloan, who first observed a notch in a foveal threshold spectrum. We have examined suprathreshold discriminability on both sides of the Sloan notch produced by a 6700 td, 578 nm adapting field. In a temporal two-alternative forced-choice paradigm, a suprathreshold 650 nm low-frequency "standard" stimulus was paired with low-frequency "test" stimuli, of wavelength between 600 and 670 nm and varied intensity; the observer's task was to identify the interval containing the standard. Discriminability of the test and standard typically dropped to chance for some particular test intensity, producing "indiscriminability action spectra", up to 0.7 log units above threshold. Truncated spectra (between about 530 and 560 nm) were also obtained from observers on the middle wavelength side of the Sloan notch, for a 550 nm standard. The indiscriminability action spectra of each observer were identical, up to scaling, with the observer's threshold action spectrum. Analysis of the action spectra shows that the indiscriminable stimuli are rendered equivalent at the input to a neural pathway where L- and M-cone signals converge with opposite sign. We also investigated discriminability in the spectral region containing and immediately surrounding the Sloan notch. Suprathreshold stimuli in the spectral region near the notch produce percepts that are always discriminable from 650 and 550 nm standards (and from one another), and thus we conclude that in this spectral region, perception is mediated in part by a pathway distinct from that which signals the standards. The action spectrum of this latter pathway was estimated with a variant of the discrimination procedure, and found similar to V lambda over the spectral region 575-610 nm.

Adaptation, Ocular↗

Three-dimensional ultrasound imaging. Clinical applications.

OBJECTIVE: The aim of this report is to describe the technology of three-dimensional (3-D) ultrasonic imaging and its impact on improved diagnosis and monitoring of ocular disease. DESIGN: The authors reviewed techniques for acquiring and displaying 3-D ultrasound data of the eye. PARTICIPANTS: The authors applied these techniques to representative individual cases, including a choroidal hemorrhage, a ciliary body melanoma, a ciliary body detachment, a displaced posterior chamber intraocular lens, and topographic analysis of a normal cornea. INTERVENTION: A computer-controlled motion system was used to perform very high-frequency (VHF) (50-MHz) and conventional (10-MHz) digital 3-D ultrasound data collection. The scanning system allowed digitization of ultrasound data from a series of parallel planes. The 3-D data could be manipulated interactively to obtain two-dimensional images in any plane through the scan volume. The 3-D images were constructed by volume rendering and could be positioned for viewing from a variety of perspectives. The 3-D ultrasound parameter images representing acoustic scatterer properties were generated by spectrum analysis of digitized echo data. Color maps representing the contour and thickness of the epithelium and stroma of the central corneal were generated by digital signal processing of 3-D echo data. RESULTS: Quantitative volume measurement and biometric techniques enhanced the diagnostic and treatment planning information content in 3-D ultrasound images. The location and extent of hemorrhage and clots within the suprachoroidal space were shown with solid modeling. Volume changes in ciliary body melanoma over time were documented and 3-D ultrasound parameter image changes associated with radiation therapy observed. In ciliary body detachment, the extent of the detachment was shown. Solid modeling of a posterior chamber intraocular lens showed misplacement of the haptic in relation to the lens capsule remnants. Keratopachymetric maps showed the range and variance of thickness and local radius of curvature measurements in the cornea. CONCLUSIONS: Quantitative volume measurement and biometric tools combined with segmentation of 3-D ultrasound images improve diagnostic and treatment planning informational content of 3-D ultrasound images through improved localization of tissue structures.

Choroid Hemorrhage↗

Exploration of glycosyl hydrolase family 75, a chitosanase from Aspergillus fumigatus.

A powerful endo-chitosanase (CSN) previously described for a large scale preparation of chito-oligosaccharides (Cheng, C.-Y., and Li, Y.-K. (2000) Biotechnol. Appl. Biochem. 32, 197-203) was cloned from Aspergillus fumigatus and further identified as a member of glycosyl hydrolase family 75. We report here a study of gene expression, functional characterization, and mutation analysis of this enzyme. Gene cloning was accomplished by reverse transcription-PCR and inverse PCR. Within the 1382-bp Aspergillus gene (GenBank accession number AY190324), two introns (67 and 82 bp) and an open reading frame encoding a 238-residue protein containing a 17-residue signal peptide were characterized. The recombinant mature protein was overexpressed as an inclusion body in Escherichia coli, rescued by treatment with 5 m urea, and subsequently purified by cation exchange chromatography. A time course 1H NMR study on the enzymatic formation of chito-oligosaccharides confirmed that this A. fumigatus CSN is an inverting enzyme. Tandem mass spectrum analysis of the enzymatic hydrolysate revealed that the recombinant CSN can cleave linkages of GlcNAc-GlcN and GlcN-GlcN in its substrate, suggesting that it is a subclass I chitosanase. In addition, an extensive site-directed mutagenesis study on 10 conserved carboxylic amino acids of glycosyl hydrolase family 75 was performed. This showed that among these various mutants, D160N and E169Q lost nearly all activity. Further investigation using circular dichroism measurements of D160N, E169Q, wild-type CSN, and other active mutants showed similar spectra, indicating that the loss of enzymatic activity in D160N and E169Q was not because of changes in protein structure but was caused by loss of the catalytic essential residue. We conclude that Asp160 and Glu169 are the essential residues for the action of A. fumigatus endo-chitosanase.

Amino Acid Sequence↗