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PubMed1988

Pathogenicity of Listeria monocytogenes in comparison to other Listeria species.

In different mouse models the pathogenicity of various strains of Listeria monocytogenes and of other Listeria spp. was evaluated. Although quantitative differences of virulence among strains of L. monocytogenes are found, all isolates have to be regarded pathogenic unless essential virulence factors, such as protein composition of the cell wall or hemolysin production, are lacking. Besides L. monocytogenes, only Listeria ivanovii is able to multiply in the murine host, though definitely to a lesser extent. The other Listeria spp., such as Listeria innocua, Listeria welshimeri and Listeria seeligeri, have to be regarded as non-pathogenic.

Animals↗
PubMed1979

Morphogenic effect of 2-deoxy-D-arabino-hexose on Rhodosporidium toruloides.

The presence of 2-deoxy-D-arabino-hexose in the growth medium caused marked morphological changes in the cells of Rhodosporidium toruloides. The originally elongated ellipsoidal cells grew spherically in the presence of the deoxy-sugar, displayed differences in cell division and separation, and were larger than the control cells. After exhaustion of glucose from the medium the cells died, although no lysis was observed. The morphological changes were accompanied by significant alterations in the carbohydrate composition of the cell wall. The wall of R. toruloides grown in the presence of the deoxy-sugar contains higher proportions of chitin and glucan, while the relative contents of mannose and galactose polymers decreased drastically in comparison to normal cells.

Basidiomycota↗
PubMed1998

Protein secretion in phosphate-limited cultures of Bacillus subtilis 168.

The secretion of proteins from Bacillus subtilis was studied under physiologically well-defined conditions in continuous cultures at a range of specific growth rates. The kinetics of secretion was analysed by using pulse-chase and immunoprecipitation techniques that allowed both processing and release to be monitored. Growth conditions were selected that were known to lead to significant changes in the anionic polymer composition of the cell wall. Under magnesium limitation only low levels of native proteins were released into the growth medium. In contrast, much higher amounts of released protein were observed under phosphate limitation. Although synthesis of native secretory proteins appeared to be highly regulated, only minor changes in the secretion of heterologous proteins were detected. Comparable kinetics of protein release of cells grown under different conditions indicated similar cell wall permeabilities. The large changes in the amounts of released proteins were not reflected in the production of chaperones and components required fro protein secretion. The data suggest that the capacity of the secretion machinery is not a major limiting step in the export of native secretory proteins.

Bacillus subtilis↗
PubMed2006

The production of a new extracellular putative long-chain saturated polyester by smooth variants of Mycobacterium vaccae interferes with Th1-cytokine production.

Mycobacterium vaccae is of major pharmaceutical interest as an immunotherapeutic agent. Although M. vaccae 15483 ATCC(T) strain displays smooth and rough colonial morphologies on solid culture media, it is not known in which conditions M. vaccae switches from one colonial morphotype to the other or whether there are biological differences, especially immunological, between them. We have found that the change from a smooth to rough stable variant occurs spontaneously at 30 degrees C. The analysis of the composition of the cell wall in both variants showed that the smooth morphotype presents an extracellular material that has never previously been described and was identified as a long-chain saturated polyester that, interestingly, is not produced by the rough morphotype. Our results also indicate that this compound could be implicated in the spreading ability of smooth colonies. Proliferation, IFN-gamma and IL-12(p40) production by splenocyte cultures was significantly higher in mice immunised with the rough variant compared with those immunised with the smooth one. This latter finding suggests that the different colonial morphology of M. vaccae may affect the immunomodulatory effects induced from M. vaccae preparations.

Animals↗
PubMed2004

A high-resolution titrator: a new approach to studying binding sites of microbial biosorbents.

The high-resolution potentiometric titration was used as a physico-chemical method to study the acid properties of selected biosorbent materials in order to quantify the functional acidic groups for sorption and to determine their affinities by considering their partial or total ionization equilibrium reactions. The Gran's method and the Henderson-Hasselbach's equation were employed in establishing the partition of the total acidity as associated with strong, weak and very weak acidic chemical active groups. The differences in the total organic acidity (A(TO)) for the two selected types of bacteria and two mycelia revealed by this method were explained by the chemical composition of their cell walls. The total organic acidities obtained were 3.87 me g(-1) for Thiobacillus ferrooxidans, 1.31 me g(-1) for Corynebacterium glutamicum, 0.81 me g(-1) for Aspergillus niger and 2.54 me g(-1) for Rhizopus arrhizus. The links between the activity of protons and the sorption capacities of the selected bioorganic matters were established. Sorption of lead by C. glutamicum and R. arrhizus biomass indicated an optimum pH of 6. It appeared that 64% (Pb(uptake)=0.48 me g(-1)) and 38% (Pb(uptake)=0.28 me g(-1)) of A(TO) were involved during lead sorption onto R. arrhizus and C. glutamicum, respectively. The applications of titration techniques become a powerful tool for the characterization of heterogeneous materials involved in biosorption and bioremediation processes.

Aspergillus niger↗
PubMed2003

Detection of fatty acids from intact microorganisms by molecular beam static secondary ion mass spectrometry.

We report the use of a surface analysis approach, static secondary ion mass spectrometry (SIMS) equipped with a molecular (ReO(4)(-)) ion primary beam, to analyze the surface of intact microbial cells. SIMS spectra of 28 microorganisms were compared to fatty acid profiles determined by gas chromatographic analysis of transesterfied fatty acids extracted from the same organisms. The results indicate that surface bombardment using the molecular primary beam cleaved the ester linkage characteristic of bacteria at the glycerophosphate backbone of the phospholipid components of the cell membrane. This cleavage enables direct detection of the fatty acid conjugate base of intact microorganisms by static SIMS. The limit of detection for this approach is approximately 10(7) bacterial cells/cm(2). Multivariate statistical methods were applied in a graded approach to the SIMS microbial data. The results showed that the full data set could initially be statistically grouped based upon major differences in biochemical composition of the cell wall. The gram-positive bacteria were further statistically analyzed, followed by final analysis of a specific bacterial genus that was successfully grouped by species. Additionally, the use of SIMS to detect microbes on mineral surfaces is demonstrated by an analysis of Shewanella oneidensis on crushed hematite. The results of this study provide evidence for the potential of static SIMS to rapidly detect bacterial species based on ion fragments originating from cell membrane lipids directly from sample surfaces.

Bacteria↗
PubMedDate not supplied

Study on agglutinating factors from flocculent Saccharomyces cerevisiae strains.

The lectin-like theory suggest that yeast flocculation is mediated by an aggregating lectinic factor. In this study we isolated an agglutinating factor, which corresponds to lectin, from whole cells by treating the flocculent wild-type Saccharomyces cerevisiae NCYC 625 strain and its weakly flocculent mutant [rho degrees ] with EDTA and two non-ionic surfactants (Hecameg and HTAC). The dialysed crude extracts obtained in this way agglutinated erythrocytes and this hemagglutination was specifically inhibited by mannose and mannose derivatives. However, SDS-PAGE profiles showed that the three reagents had different effects on the yeast cells. The non-ionic surfactants appeared to be the most efficient, as their extracts possessed the highest specific agglutinating activity. The products released by the wild-type strain presented a higher specific agglutinating activity than those released by the [rho degrees ] mutant. Purification of the agglutinating factor from extracts of both strains by affinity chromatography revealed two active bands of relative mass of 26 and 47 kDa on SDS-PAGE. Mass spectrometry analysis by MALDI-TOF, identified a 26 kDa band as the triose phosphate isomerase (TPI) whereas a 47 kDa band was identical to enolase. Edman degradation showed that the N-terminal sequences of these proteins were similar to TPI and enolase, respectively. The difference in the flocculation behaviour of the two strains is due to changes in the protein composition of the cell wall and in the protein structure involved in cell-cell recognition.

Agglutinins↗
PubMed1994

The influence of low growth temperature on the amount of free R lipopolysaccharide, on the expression of R-core determinants and on O-chain lengths in Salmonella S forms.

Ten Salmonella strains belonging to five serological groups were cultivated both at 37 degrees C and close to their individual temperature minima at 12-14 degrees C and the composition of their cell wall lipopolysaccharides (LPS) was compared. When grown at low temperature, the proportion of unsubstituted (free) R-LPS in the total LPS moiety increased significantly in 6 strains, whereas in the other strains, no change or even a slight decrease in the R-LPS proportion was observed as judged from the analyses by SDS-PAGE. In the immunoblot, the R-LPSs from 9 out of 10 strains showed a modified reactivity against a set of specific Salmonella R antisera (anti-Ra, anti-Rb1, anti-Rb2, anti-Rc). In most cases, the decrease in Rb1 reactivity was paralleled by an increase in Rb2 reactivity and also by an increase in the total amount of free R-LPS. The electrophoretic mobility of free R-LPS was changed in 7 out of 10 strains, although the changes were not unidirectional. All changes occurred only in the range of the Ra-Rb1 chemotypes and no significant correlation to the serological grouping of the strains was evident. When grown at low temperature, the average number of O-repeating units was reduced in the majority of cases and in some cases, also the banding profile in SDS-PAGE in the S-LPS region was modified. The fatty acid spectra showed some changes which were in accordance with previous results, namely a decrease in the content of C-12:0 and C-16:0 and an increase in that of C-14:0 and C-16:1. The different influences of the growth temperature on the LPS biosynthesis of different Salmonella strains may be a result of the genetic diversity of this group of microorganisms.

Electrophoresis, Polyacrylamide Gel↗
PubMed2002

The pathobiology of Paracoccidioides brasiliensis.

Paracoccidioides brasiliensis causes one of the most prevalent systemic mycoses in Latin America--paracoccidioidomycosis. It is a dimorphic fungus that undergoes a complex transformation in vivo, with mycelia in the environment producing conidia, which probably act as infectious propagules upon inhalation into the lungs, where they transform to the pathogenic yeast form. This transition is readily induced in vitro by temperature changes, resulting in modulation of the composition of the cell wall. Notably, the polymer linkages change from beta-glucan to alpha-glucan, possibly to avoid beta-glucan triggering the inflammatory response. Mammalian oestrogens inhibit this transition, giving rise to a higher incidence of disease in males. Furthermore, the susceptibility of individuals to paracoccidioidomycosis has a genetic basis, which results in a depressed cellular immune response in susceptible patients; resistance is conferred by cytokine-stimulated granuloma formation and nitric oxide production. The latency period and persistence of the disease and the apparent lack of efficacy of humoral immunity are consistent with P. brasiliensis existing as a facultative intracellular pathogen.

Amino Acid Sequence↗
PubMed2002

Phenotypic switching in Cryptococcus neoformans.

Cryptococcus neoformans strains exhibit considerable phenotype variability with regards to the capsular polysaccharide, sterol composition of the cell wall, and cell and colony morphology. Phenotypic changes can occur spontaneously during in vitro passage of strains or during chronic infection in vivo and may be associated with differences in virulence. Studies from our laboratory have demonstrated that phenotype variability can be the result of phenotypic switching. Phenotypic switching is defined as a reversible change of an observable colony phenotype that occurs at a frequency above the expected frequency for somatic mutations. This implies that phenotypic switching represents controlled and programmed changes in this pathogenic yeast rather than random mutations. We have shown that a phenotypic switch from a smooth colony phenotype to a mucoid colony phenotype occurs in vitro and in vivo during chronic infection of mice. More importantly we have now demonstrated that the switch is associated with an increase in virulence and a change in the host immune response. Implications of these findings for the pathogenesis of cryptococcosis are discussed.

Cryptococcosis↗
PubMed2003

Thermal stability of texture in Chinese water chestnut may be dependent on 8,8'-diferulic acid (aryltetralyn form).

Ferulic acid (FA) cross-links have been implicated in the thermal stability of texture in Chinese water chestnut (CWC) tissues. The aim of the current study has been to investigate this concept further. CWC tissue strips were measured for their mechanical properties before and after extraction in increasing strengths of alkali. The mechanical properties were related to the associated mode of fracture (cell separation or breakage) at the fracture surfaces and the phenolic composition of the cell walls. CWC tissue softened after prolonged extraction in cold alkali due to an increase in the ease of cell separation. Analysis of wall-bound phenolics demonstrated that most FA moieties, including five of the six dehydrodimers, were released before tissue strength was reduced. Loss of strength was, however, coincident with the loss of 8,8'-diferulic acid, aryltetralin (AT) form. It has been suggested that this dehydrodimer may be particularly concentrated at the edge of the cell faces. These results provide further evidence for the involvement of this dehydrodimer in conferring thermal stability of cell-cell adhesion and hence texture in CWC. However, they do not exclude the other diferulates from involvement in cell adhesion.

Biomechanical Phenomena↗
PubMed2006

Effect of industrial dehydration on the soluble carbohydrates and dietary fiber fractions in legumes.

The effects of soaking, cooking, and industrial dehydration treatments on soluble carbohydrates, including raffinose family oligosaccharides (RFOs), and also on total dietary fiber (TDF), insoluble dietary fiber (IDF), and soluble (SDF) dietary fiber fractions were studied in legumes (lentil and chickpea). Ciceritol and stachyose were the main alpha-galactosides for chickpea and lentil, respectively. The processing involved a drastic reduction of soluble carbohydrates of these legumes, 85% in the case of lentil and 57% in the case of chickpea. The processed legume flours presented low residual levels of alpha-galactosides, which are advisable for people with digestive problems. Processing of legumes involved changes in dietary fiber fractions. A general increase of IDF (27-36%) due to the increase of glucose and Klason lignin was observed. However, a different behavior of SDF was exhibited during thermal dehydration, this fraction increasing in the case of chickpea (32%) and decreasing in the case of lentil (27%). This is probably caused by the different structures and compositions of the cell wall networks of the legumes.

Carbohydrates↗
PubMed2002

Influence of the growth substrate on the mycolic acid profiles of mycobacteria.

The influence of growth substrates on mycolic acid profiles of PAH-degrading Mycobacterium spp. LB501T, LB307T and VM552 was examined by high-performance liquid chromatography (HPLC) using glucose, alkanes, polycyclic aromatic hydrocarbons (PAH) or Luria-Bertani medium (LB) as the sole carbon source. The substrates gave rise to varying mycolic acid profiles, as bacteria growing on poorly water-soluble substrates exhibited more hydrophobic mycolic acids than cells grown on glucose. Our results indicate that mycobacteria respond to the growth substrate by changing the mycolic acid composition of their cell wall, pointing at the importance of the growth substrate for mycolic acid profiling as an identification method of actinomycetes.

Biodegradation, Environmental↗
PubMed2000

Pectin engineering: modification of potato pectin by in vivo expression of an endo-1,4-beta-D-galactanase.

Potato tuber pectin is rich in galactan (oligomer of beta-1,4-linked galactosyl residues). We have expressed a fungal endo-galactanase cDNA in potato under control of the granule bound starch synthase promoter to obtain expression of the enzyme in tubers during growth. The transgenic plants displayed no altered phenotype compared with the wild type. Fungal endo-galactanase activity was quantified in the transgenic tubers, and its expression was verified by Western blot analysis. The effect of the endo-galactanase activity on potato tuber pectin was studied by Fourier transform infrared microspectroscopy, immuno-gold labeling, and sugar analysis. All analyses revealed alterations in pectin composition. Monosaccharide composition of total cell walls and isolated rhamnogalacturonan I fragments showed a reduction in galactosyl content to 30% in the transformants compared with the wild type. Increased solubility of pectin from transgenic cell walls by endo-polygalacturonase/pectin methylesterase digestion points to other changes in wall architecture.

Genetic Engineering↗
PubMedDate not supplied

Some observations on the pneumococcus and on the current status of pneumococcal disease and its prevention.

Although the means whereby pneumococcus damages the hosts it invades remain unknown, some advances in understanding the biology of this organism and the pathogenesis of pneumococcal infection have been made in recent years. Knowledge of the composition of the cell wall (C) polysaccharide of the pneumococcus and its relation to the capsular polysaccharides of this organism is considered and a hypothesis of the evolution of the pneumococcal capsule is entertained. Refinements in understanding the pathogenesis of pneumococcal pneumonia, bacteremia, and otitis media are reviewed, and current knowledge of the epidemiology of pneumococcal infections and of the distribution of pneumococcal types causing such infections is discussed. The history of the development of pneumococcal vaccines is summarized briefly, together with an account of the field trials of contemporary polyvalent vaccines of pneumococcal capsular polysaccharides conducted recently.

Antigens, Bacterial↗
PubMed2005

Biological consequences of petite mutations in Candida glabrata.

OBJECTIVES: To define the pathogenicity of respiration-deficient mutants of Candida glabrata, which present a reduced susceptibility to azoles, that are easily induced in vitro by exposure to these drugs or to ethidium bromide and that may be selected in vivo in patients receiving fluconazole. METHODS: Two wild-type isolates of C. glabrata were compared with their respective fluconazole- or ethidium bromide-induced petite mutants, regarding the carbohydrate and protein composition of the cell wall, as well as their surface physical properties, and also their adherence abilities and virulence in mice. RESULTS: Flow cytometric analysis of cell wall carbohydrates using several fluorescent lectins showed an increased binding of mutant cells to concanavalin A compared with their parent isolates, suggesting a greater availability or an increased amount of glucose-mannose residues at the cell surface in petite mutants. Likewise, some quantitative differences between parent and mutant isolates were shown by SDS-PAGE in protein extracts from blastoconidia. Regarding the surface physical properties, no significant differences were seen in the electrophoretic mobility determined by microelectrophoresis, but the two-phase partitioning method revealed a lower cell surface hydrophobicity for petite mutants. Moreover, mutant cells exhibited significant overexpression of CgEPA1 as revealed by real-time reverse transcription-PCR, but the adherence capacities to Caco-2 cells, a human enterocyte line, were not significantly different. Finally, in agreement with their slower growth, petite mutants were less virulent than parent isolates in a murine model of systemic infection. CONCLUSION: This low virulence in mice suggests that petite mutants could be disregarded clinically although they may arise during fluconazole therapy.

Animals↗
PubMed2000

Uptake and retranslocation of leaf-applied cadmium (109Cd) in diploid, tetraploid and hexaploid wheats.

Uptake and retranslocation of leaf-applied radiolabeled cadmium (109Cd) was studied in three diploid (Triticum monococcum, AA), four tetraploid (Triticum turgidum, BBAA) and two hexaploid (Triticum aestivum, BBAADD) wheat genotypes grown for 9 d under controlled environmental conditions in nutrient solution. Among the tetraploid wheats, two genotypes were primitive (ssp. dicoccum) and two genotypes modern wheats (ssp. durum). Radiolabelled Cd was applied by immersing the tips (3 cm) of mature leaf into a 109Cd radiolabelled solution. There was a substantial variation in the uptake and export of 109Cd among and within wheat species. On average, diploid wheats (AA) absorbed and translocated more 109Cd than other wheats. The largest variation in 109Cd uptake was found within tetraploid wheats (BBAA). Primitive tetraploid wheats (ssp. dicoccum) had a greater uptake capacity for 109Cd than modern tetraploid wheats (ssp. durum). In all wheats studied, the amount of the 109Cd exported from the treated leaf into the roots and the remainder of the shoots was poorly related to the total absorption. For example, bread wheat cultivars were more or less similar in total absorption, but differed greatly in the amount of 109Cd retranslocated. The diploid wheat genotype 'FAL-43' absorbed the lowest amount of 109Cd, but retranslocated the greatest amount of 109Cd in roots and remainder of shoots. The results indicate the existence of substantial genotypic variation in the uptake and retranslocation of leaf-applied 109Cd. This variation is discussed in terms of potential genotypic differences in binding of Cd to cell walls and the composition of phloem sap ligands possibly affecting Cd transport into sink organs.

Biological Transport↗
PubMed1996

Bartonella vinsonii subsp. berkhoffii subsp. nov., isolated from dogs; Bartonella vinsonii subsp. vinsonii; and emended description of Bartonella vinsonii.

Two bacterial strains, one isolated from the blood of a dog with valvular endocarditis and one isolated from the blood of a healthy dog, were similar to Bartonella species, as determined by a number of phenotypic criteria, including growth characteristics, biochemical reactions, and cell wall fatty acid composition. The results of 16S rRNA gene sequence similarity studies confirmed that these strains are closely related and belong in the genus Bartonella and that Bartonella vinsonii is their closest relative (the 16S rRNA of isolate 93-C01T [T = type strain] was 99.37% identical to the 16S rRNA of the type strain of B. vinsonii, the 16S rRNA of isolate G7464 was 99.61% identical to the 16S rRNA of the type strain, and the 16S rRNAs of the dog isolates were 99.77% identical to each other). The 16S rRNAs of both strains contained a 12-base insertion that was not present in the 16S rRNA of the type strain of any Bartonella species. DNA relatedness tests revealed that these strains were related at the species level to the type strain of B. vinsonii. They were, however, significantly more closely related to each other than to B. vinsonii. On the basis of their unique 16S rRNA sequence insertion, their preferentially high level of relatedness, and their similar origins (dogs), we believe that strains 93-C01(T) and G7464 should be placed in a separate subspecies of B. vinsonii, for which we propose the name B. vinsonii subsp. berkhoffii subsp. nov. The type strain of B. vinsonii subsp. berkhoffii is strain 93-C01 (= ATCC 51672). The description of B. vinsonii is emended to accommodate the new subspecies, and B. vinsonii subsp. vinsonii is described.

Animals↗