Structural studies of the capsular polysaccharide of a non-neoformans Cryptococcus species identified as C. laurentii, which was reclassified as Cryptococcus flavescens, from a patient with AIDS

Cryptococcus flavescens, a strain originally identified as C. laurentii, was isolated from the cerebrospinal fluid of an AIDS patient, and the soluble capsular polysaccharide of the yeast was investigated. Glucuronoxylomannan (GXM) was obtained from C. flavescens under conditions similar to those used to obtain C. neoformans polysaccharide. However, the GXM differed from C. neoformans polysaccharide in the decreased O-acetyl group content. The structure of GXM was determined by methylation analysis, partial acid hydrolysis, NMR analyses, and controlled Smith degradation. These analyses indicated that GXM has the following structure: an alpha-(1-->3)-D-mannan backbone with side chains of beta-D-glucuronic acid residues bound to the C-2 position of the mannose residue. The C-6 position of the mannose is substituted with D-man-beta-(1-->4)-D-xyl-beta-(1--> disaccharide. Furthermore, the existence of side chains containing more than two xylose residues was suggested. This mannosylxylose side chain is a novel structure in polysaccharides of C. neoformans and other Cryptococcus species.     

Studies on projections of somatic afferentation in the neocortex of the hedgehog Erinaceus europaeus]

Cortical projections of the somatic system in the hedgehog cortex were studied by means of the evoked potential (EP) technique. The EPs were shown to arise at a large area of the lateral cortex, the area in question being enlarged under chloralose anaesthesia. In the focus of projection of corresponding modalities the EPs posessed the least latency. Functional differences in the EP generation system both in the focus of projection and outside it are discussed. The wide EPs spreading in the lateral cortex as well as the presence of relatively differentiated projections is supposed to depend on the afferent volley dispersion to the polysynaptic pathways at the thalamic level. Considerable overlapping of sensory projection zones in the cortex is conditioned by the latter and indicates low degree of the somatic analyser organization.     

GOurmet: A tool for quantitative comparison and visualization of gene expression profiles based on gene ontology (GO) distributions

Background  

The ever-expanding population of gene expression profiles (EPs) from specified cells and tissues under a variety of experimental conditions is an important but difficult resource for investigators to utilize effectively. Software tools have been recently developed to use the distribution of gene ontology (GO) terms associated with the genes in an EP to identify specific biological functions or processes that are over- or under-represented in that EP relative to other EPs. Additionally, it is possible to use the distribution of GO terms inherent to each EP to relate that EP as a whole to other EPs. Because GO term annotation is organized in a tree-like cascade of variable granularity, this approach allows the user to relate (e.g., by hierarchical clustering) EPs of varying length and from different platforms (e.g., GeneChip, SAGE, EST library).     

Effects of trehalose on stress tolerance and biocontrol efficacy of Cryptococcus laurentii

AIMS: To investigate the effects of internal trehalose on viability and biocontrol efficacy of antagonistic yeast Cryptococcus laurentii under stresses of low temperature (LT), controlled atmosphere (CA) and freeze drying. METHODS AND RESULTS: The content of trehalose in C. laurentii was increased by culturing the yeast in trehalose-containing medium. Compared with yeast cells with low trehalose level, the yeast cells with high level of internal trehalose not only obtained higher viability, but also showed higher population and better biocontrol efficacy against Penicillium expansum on apple fruit both at 1 degrees C and in CA condition (5% O(2), 5% CO(2), 1 degrees C). After freeze drying, survival of the yeast with high trehalose level was markedly increased when stored at 25 degrees C for 0, 15 and 30 days. Meanwhile, high integrity of plasma membrane was detected in the freeze-dried yeast with high trehalose level by propidium iodide staining. CONCLUSIONS: Induced accumulation of internal trehalose could improve viability and biocontrol efficacy of C. laurentii under stresses of LT and CA. Moreover, survival of the yeast was also increased as internal trehalose accumulation after freeze drying, and one of the reasons might be that trehalose gave an effective protection to plasma membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this experiment show a promising way to improve the biocontrol performance of antagonistic yeasts under the commercial conditions.     

Prostaglandin E receptors as inflammatory therapeutic targets for atherosclerosis

Prostaglandin E receptors (EPs) are the G-protein-coupled receptors (GPCRs) that respond to type E(2) prostaglandin (PGE(2)). Data has shown that PGE(2) may function as an endogenous anti-inflammatory mediator by suppressing the production of cytokines. However, other studies have demonstrated that PGE(2), a pro-inflammatory mediator produced by various cell types within the wounded vascular wall, plays a crucial role in early atherosclerotic development. These contradictory results may be due to the versatility of EPs. Experimental data suggest an individual role for each PGE(2) receptor, such as EP(1), EP(2), EP(3) and EP(4), in atherosclerosis. In this review, the roles of EPs in atherosclerosis are summarized, and the value of EPs as new therapeutic targets for atherosclerosis is explored.     

Conformation and cooperative order-disorder transition in aqueous solutions of β-1,3-d-glucan with different degree of branching varied by the Smith degradation

β-1,3-d -glucan with different degrees of branching were obtained by selectively and gradually removing side chains from schizophyllan, a water-soluble triple helical polysaccharide, using the Smith degradation. Size exclusion chromatography combined with a multi-angle light scattering detection was performed in aqueous 0.1 M NaCl. The degree of branching decreased after the Smith degradation, while the molar mass distributions were almost unchanged. The molecular conformation of the Smith-degraded β-1,3-d -glucan was analyzed on the basis of the molar mass dependency of the radius gyration, and found to be comparable to the original triple helix of schizophyllan. Differential scanning calorimetry in deuterium oxide–hexadeuterodimethylsulfoxide mixtures was performed to investigate the effects of the degree of branching on the cooperative order-disorder transition. Removal of side chains affects both the transition temperature and transition enthalpy. The ordered structure is formed by the residual side chains in the triplex unit, so that the linear cooperative system of the triplex is maintained after the Smith degradation.     

Evoked activity of the cat hypothalamus and amygdala under food motivation and in emotional stress

Amplitude-latency characteristics of auditory evoked potentials (EPs) recorded in bilateral points of the lateral hypothalamus and amygdala were studied under food motivation, in emotional stress (presentation of dogs) and tentative reactions. In the state of hunger, as compared with safety, the latencies of P1, N2 components of EP in hypothalamus, and P1, N2, N3 in amygdala were decreased and their amplitudes were changed. Changes in the left side of both structures were more pronounced. During presentation of dogs, decreases of latencies of all EP components including N1 occurred in hypothalamus and amygdala, changes in hypothalamic potentials were more pronounced on the right side, whereas in the amygdala--on the left side. During tentative responses to emotional-neutral stimuli, the latency of EP increased. It was concluded that sensory reactivity of hypothalamus and amygdala increased in motivational-emotional states. It was supposed that the side of dominance of structure may be related both to the factors of active or passive behavior during fear and the genesis of emotion (motivational or informational).     

Some chemical and physical properties of extracellular polysaccharides produced by Butyrivibrio fibrisolvens strains

Most strains of Butyrivibrio fibrisolvens are known to produce extracellular polysaccharides (EPs). However, the rheological and functional properties of these EPs have not been determined. Initially, 26 strains of Butyrivibrio were screened for EP yield and apparent viscosities of cell-free supernatants. Yields ranged from less than 1.0 to 16.3 mg per 100 mg of glucose added to the culture. Viscosities ranged from 0.71 to 5.44 mPa.s. Five strains (CF2d, CF3, CF3a, CE51, and H10b) were chosen for further screening. The apparent viscosity of the EP from each of these strains decreased by only 50 to 60% when the shear rate was increased from 20 to 1,000 s-1. Strain CE51 produced the EP having the highest solution viscosity. A detailed comparison of shear dependency of the EP from strain CF3 with xanthan gum showed that this EP was less shear sensitive than xanthan gum and, at a shear rate of 1,000 s-1, more viscous. EPs from strains CF3 and H10b were soluble over a wide range of pH (1 to 13) in 80% (vol/vol) ethanol-water or in 1% (wt/vol) salt solutions. The pH of 1% EP solutions was between 4.5 and 5.5. Addition of acid increased solution viscosities, whereas addition of base decreased viscosity. EPs from strains CF3, CE51, and H10b displayed qualitatively similar infrared spectra. Calcium and sodium were the most abundant minerals in the three EPs. The amounts of magnesium, calcium, and iron varied considerably among the EPs, but the potassium contents remained relatively constant.     

Some chemical and physical properties of extracellular polysaccharides produced by Butyrivibrio fibrisolvens strains.

Most strains of Butyrivibrio fibrisolvens are known to produce extracellular polysaccharides (EPs). However, the rheological and functional properties of these EPs have not been determined. Initially, 26 strains of Butyrivibrio were screened for EP yield and apparent viscosities of cell-free supernatants. Yields ranged from less than 1.0 to 16.3 mg per 100 mg of glucose added to the culture. Viscosities ranged from 0.71 to 5.44 mPa.s. Five strains (CF2d, CF3, CF3a, CE51, and H10b) were chosen for further screening. The apparent viscosity of the EP from each of these strains decreased by only 50 to 60% when the shear rate was increased from 20 to 1,000 s-1. Strain CE51 produced the EP having the highest solution viscosity. A detailed comparison of shear dependency of the EP from strain CF3 with xanthan gum showed that this EP was less shear sensitive than xanthan gum and, at a shear rate of 1,000 s-1, more viscous. EPs from strains CF3 and H10b were soluble over a wide range of pH (1 to 13) in 80% (vol/vol) ethanol-water or in 1% (wt/vol) salt solutions. The pH of 1% EP solutions was between 4.5 and 5.5. Addition of acid increased solution viscosities, whereas addition of base decreased viscosity. EPs from strains CF3, CE51, and H10b displayed qualitatively similar infrared spectra. Calcium and sodium were the most abundant minerals in the three EPs. The amounts of magnesium, calcium, and iron varied considerably among the EPs, but the potassium contents remained relatively constant.     

Electrophysiologic analysis of the process of recognizing target and non-target stimuli in healthy and oligophrenic children

In an automatized experiment, with a computer on line, amplitude-temporal parameters of evoked potentials (EPs) to purposive and non-purposive stimuli (digits), were analyzed in normal and mental retarded children. At unilateral stimuli presentation to the left or right visual half-fields EPs were recorded simultaneously in projection, TPO, parietal and central areas of the left and right hemispheres. It has been shown that in normal children, differential involvement of projection and associative structures in the analysis of sensory information takes place in both hemispheres. The amplitudes of most EP components in the range of 100-400 ms to the purposive stimuli are higher than to the non-purposive ones. Considerable similarity of EPs developing in response to ipsi- and contralateral stimulations of visual fields ("direct" and "transmitted" EP) is observed. In mental retarded children significant changes are revealed in intra- and interhemisphere organization of the process of perception of purposive and non-purposive stimuli. In the right hemisphere structures there are no differential EP reactions to the two types of stimuli. Significant, in comparison with the norm, prolongation of the latencies of most EP components is noted, especially in the structures of the left hemisphere, to the purposive stimuli. In the process of perception, changes are seen of the integration of functions of both hemispheres. The totality of disturbances of systemic brain organization at perceptive activity in mental retarded children may reflect neurophysiological mechanisms of mental deficiency.     

Structural transformations of arabinogalactan from the Siberian larch during mechanochemical processing and the biological properties of the products

It has been found by gel-penetrating chromatography and quantitative ¹³C NMR spectroscopy that mechanochemical processing of arabinogalactan isolated from the wood of the Siberian larch changes the molecular mass distribution, monosaccharide composition, and the degree of branching of its macromolecules. This is due to the partial destruction of polysaccharide macromolecules and subsequent recombination of resulting fragments. The intensity of these processes depends on the conditions of mechanochemical processing. Based on the data of IR and ¹³C NMR spectra, the conclusion has been drawn that no functionalization of arabinogalactan macromolecules occurs under these conditions. The toxicopharmacological properties of mechanically processed arabinogalactan have been studied. Its acute toxicity is >5000 mg/kg.     

Impact of Anatomic Site on Growth,Efflux-Pump Expression,Cell Structure,and Stress Responsiveness of <Emphasis Type="Italic">Bacteroides fragilis</Emphasis>

This study investigated whether B. fragilis from various human sites acquired stable traits enabling it to express certain efflux pumps (EPs), adopt a particular cell structure, and tolerate certain stressors. Isolates from blood, abscess, and stool (n = 11 each) were investigated. Bacteria from various sites portrayed different ultrastructres and EP expression. Blood isolates were tolerant to nutrient limitation and stool isolates to NaCl and bile salt stress. Stressors significantly increased EP expression. These data demonstrate that (1) B. fragilis acquires stable traits from various in vivo microenvironments; (2) that EPs are involved in stress responsiveness; and (3) that EP expression is tightly controlled and site dependent.     

Effect of pH on the molecular weight of poly-3-hydroxybutyric acid produced by Alcaligenes sp.

Summary The molecular weight of a polymer is an important parameter characterizing the physical properties of the polymer. The effect of pH on the molecular weight of poly-3-hydroxybutyric acid (PHB) produced by Alcaligenes sp. K-912 were investigated. PHB having higher molecular weight was obtained when cultivated under high pH conditions compared to the results cultivated under low pH conditions. The polydispersity index was almost the same as 1.6. But the cell mass was decreased as the pH became higher. The molecular weight was mainly determined in the early cultivation period and molecular weight difference could be explained by calculating the number of polymer chains.     

Partial muting leads to age-dependent modification of motor patterns underlying crystallized zebra finch song

The crystallized structure of adult zebra finch (Taeniopygia guttata) song is modifiable if sensory feedback is altered during sound production. Such song plasticity has been studied by examining acoustic modifications to the motif; however, the underlying changes to the vocal motor patterns of these acoustic modifications have not been addressed. Adult birds in two age categories (young=90-120 days or middle aged 150-250 days) that sang crystallized song were used in the experiment. Vocal motor patterns were monitored by recording respiratory air sac pressure before, during, and after song plasticity was induced by partial or complete reduction of phonation (i.e., "partial muting"). Birds were recorded until changes in air sac pressure patterns underlying the song structure were observed (up to 160 days). Young adult birds were more likely to insert shorter duration (<125 ms) expiratory pulses (EPs) into the motif than middle-aged adults. These shorter duration EPs were produced with a unique pressure pattern relative to the intact song, and therefore appeared to be generated by novel motor gestures. Stuttering (atypical repetition of an EP) was observed when these novel EPs were inserted into the motif, regardless of age. The EP of the distance call, which is also a learned vocalization in zebra finches, showed a similar reduction in duration if EPs were also shortened in the song. The emergence of shorter duration EPs was not related to sound production, or nonspecific effects of the surgical procedure, which suggests an age-dependent neural process for song plasticity.     

Effect of oxygen and shear stress on molecular weight of hyaluronic acid

Dissolved oxygen (DO) and shear stress have pronounced effects on hyaluronic acid (HA) production, yet various views persist about their effect on the molecular weight of HA. Accordingly, this study investigated the effects of DO and shear stress during HA fermentation. The results showed that both cell growth and HA synthesis were suppressed under anaerobic conditions, and the HA molecular mass was only (1.22+/-0.02) x 106 Da. Under aerobic conditions, although the DO level produced no change in the biomass or HA yield, a high DO level favored the HA molecular mass, which reached a maximum value of (2.19+/- 0.05) x 106 Da at 50% DO. Furthermore, a high shear stress delayed the rate of HA synthesis and decreased the HA molecular weight, yet had no clear effect on the HA yield. Therefore, a high DO concentration and mild shear environment would appear to be essential to enhance the HA molecular weight.     

Cerebral evoked potentials after rectal stimulation

We obtained reproducible cortical evoked potentials (EPs) in response to electrical stimulation of the rectum with 1 Hz frequency. We found 2 distinctly different EPs in response to rectal stimulation. In 5 females, the EP had an early onset latency (mean 26 msec) with multiple positive and negative peaks. In 10 females, the EP had a later onset latency (mean 52 msec) and a trifid configuration, having a very prominent negative peak. The early onset EPs after rectal stimulation appeared very similar to the wave form of the cortical EPs recorded after pudendal nerve stimulation. Finding similar interpeak latencies in the early onset EP after rectal stimulation and the EP after pudendal nerve stimulation suggests that either the same pathway was used or that rectal stimulation also stimulated the pudendal nerve. It appears that we stimulated visceral afferents when we recorded late onset EPs, because the large EP amplitude declined rapidly with faster stimulation rates and also with greater number of averaging, and the sensation threshold was very unstable, all different to somatosensory EPs.     

Manifestations of the genotypic causality of human evoked potentials during perception of various visual stimuli

Intrapair resemblance of the wave form and amplitude-temporal parameters of evoked potentials (EPs) to flashes, chess field, house image, the word "house" and a series of other stimuli was evaluated in 20 pairs of monozygotic and 20 pairs of homosexual dizygotic adult twins. In the occipital area the maximum of genetic dependence was characteristic of EPs to flashes, the minimum--of EPs--to the word "house". In vertex EPs parameters genotypic effects were manifest irrespectively of the stimulus type. Genotypic dependence differed for the amplitudes and latencies of separate EP components.     

Mechanistic investigation of a starch-branching enzyme using hydrodynamic volume SEC analysis

Two linear alpha-(1,4)-D-glucans substrates, of degrees of polymerization DP approximately 150 and 6000, were exposed to maize starch-branching enzyme IIa (mSBEIIa) in vitro. The resulting branched alpha-glucans and their constituent chains (obtained by debranching) were analyzed by nuclear magnetic resonance (NMR) and size-exclusion chromatography (SEC). SEC data for the debranched species are presented as chain-length distributions, while those for branched species are presented as hydrodynamic volume distributions (HVDs), which is the most meaningful way to present such data (because SEC separates by size, not molar mass, and a sample of branched polymers with the same size can have a range of molar masses). A rigorous interpretation of the HVDs of the substrate and its branched product show that at least part of the branching is an interchain transfer mechanism in both the short- and long-chain substrate cases. A bimodal HVD of the in vitro branched alpha-glucan derived from the short-chain substrate was observed, and it is postulated that the divergence of the two populations is due to very small chains being unable to undergo branching. In the case of the in vitro branching of the long-chain substrate, the formation of maltohexaose during the reaction and the presence of a monomodal HVD were observed, suggesting a distinct mode of action of mSBEIIa on this substrate. Quantification of the branching level by NMR showed the branched glucans from both substrates had substantial amounts of branching (2.1-4.5%), ascribed to the intrinsic nature of the action of mSBEIIa on the two substrates. It is postulated that differences in the degrees of substrate association affect the pattern of branching catalyzed by the enzyme, and a putative active site structure is proposed based on the appearance of maltohexaose. The molar mass distribution of the constituent chains of the in vitro branched alpha-glucans obtained by isoamylase treatment reveals the transfer of chains of specific size and supports the supposition given in the literature that mSBEIIa is responsible for short-chain branching in amylopectin. It is suggested that hydrodynamic volume SEC analysis should be used as a tool for the mechanistic investigation of SBEs, allowing SEC data of in vitro branched alpha-glucans to be both comparable and quantitative.