Does water activity rule P. mirabilis periodic swarming? II. Viscoelasticity and water balance during swarming

Following the analysis of the biochemical and functional properties of the P. mirabilis extra cellular matrix performed in the first part of this study, the viscoelasticity of an actively growing colony was investigated in relation to water activity. The results demonstrate that the P. mirabilis colony exhibits a marked viscoelastic character likely due to both cell rafts and exoproduct H-bond networks. Besides, the water loss by evaporation during migration has been measured, whereas the experimental determination of the water diffusion coefficient in agar has allowed us to estimate the net water influx at the agar/colony interface. These data drive us to propose that a periodic increase of the water activity at the colony's periphery, mainly due to the drastic surface to volume ratio increase associated with swarming, causes the periodic and synchronous cessation of migration through the dissociation of exoproduct networks, which in turn strongly alters the matrix viscoelasticity.     

Electrophoretic Characteristics of Staphylococcal Hyaluronate Lyase

Partially purified staphylococcal hyaluronidase was studied with respect to its electrophoretic properties by the use of starch slurry or semisolid Noble agar as a matrix. The polarity of enzyme activity was found to be cathodal on starch and anodal on agar gel. The electrophoretic migration of the partially purified enzyme on starch, as a function of pH, suggested that the enzyme has an isoelectric point of pH 9.5 to 10.     

Immobilization of methane-oxidizing bacterial cells

The purpose of this work was to study how the conditions for immobilizing the cells of the methane oxidizing bacterium Methylomonas rubra using chemical and physical techniques influence their functional catalytical properties. These properties were found to depend on the chemical nature of a matrix and the mode of binding the cells to the carrier. The best carriers were shown to be silochrome with introduced carboxy groups and agar gel.     

Serum protease inhibitors promote anchorage-independent growth of transformed cells in serum-free medium.

The effect of three serum serine protease inhibitors on the serum-free agar growth of an SV40-transformed 3T3 cell line was investigated. Antithrombin III, alpha-2-macroglobulin and alpha-1-antitrypsin were found to potently stimulate colony growth in a semisolid medium because of their anti-proteolytic properties. These results indicate that protease inhibitors can facilitate tumor cell growth in serum-free agar cultures and suggest that the stimulatory effect of serum on the growth of certain transformed cells in agar may at least partially be due to the high levels of protease inhibitors found in serum.     

Thrombospondins in the heart: potential functions in cardiac remodeling

Cardiac remodeling after myocardial injury involves inflammation, angiogenesis, left ventricular hypertrophy and matrix remodeling. Thrombospondins (TSPs) belong to the group of matricellular proteins, which are non-structural extracellular matrix proteins that modulate cell–matrix interactions and cell function in injured tissues or tumors. They interact with different matrix and membrane-bound proteins due to their diverse functional domains. That the expression of TSPs strongly increases during cardiac stress or injury indicates an important role for them during cardiac remodeling. Recently, the protective properties of TSP expression against heart failure have been acknowledged. The current review will focus on the biological role of TSPs in the ischemic and hypertensive heart, and will describe the functional consequences of TSP polymorphisms in cardiac disease.     

Influence of growth environment on coaggregation between freshwater biofilm bacteria

AIM: To characterize the expression of coaggregation between Blastomonas natatoria 2.1 and Micrococcus luteus 2.13 following growth in liquid culture, on agar and in an artificial biofilm matrix composed of poloxamer hydrogel. METHODS AND RESULTS: The ability of B. natatoria 2.1 and M. luteus 2.13 to coaggregate with one another was assessed following growth in liquid culture as colonies on agar or within a poloxamer hydrogel matrix. In all these environments a cycle of gain and loss of coaggregation occurred when the two cell types were aged simultaneously, with optimum expression occurring in early stationary phase. Blastomonas natatoria 2.1 cells only coaggregated maximally after entry into stationary phase. Conversely, M. luteus 2.13 cells only coaggregated in exponential phase and early stationary phase and coaggregation ability was lost in late stationary phase. Maximal coaggregation therefore only occurred between the two strains if both were in early stationary phase, when the surface properties of the two cell types were optimal for coaggregation. CONCLUSION: In addition to occurring between cells grown in liquid culture, coaggregation between aquatic bacteria occurs after growth as a biofilm on agar and in an artificial biofilm matrix in poloxamer. Under all conditions, the B. natatoria 2.1 coaggregation adhesin and complementary receptor on M. luteus 2.13 were only expressed simultaneously during early stationary phase.     

Comparative studies on the conformational change and aggregation behavior of irradiated carrageenans and agar by dynamic light scattering

The conformational associative properties of kappa-, iota-, and lambda-carrageenan and agar with irradiation dose were studied by dynamic light scattering. The random scission of the carrageenans and agar by gamma irradiation resulted in the formation of polydispersed lower molecular weight fragments. At high doses, the system moves towards uniformity. Conformational change from coil to helix was observed in all carrageenans and agar at doses up to 100 kGy. The conformational change in lambda-carrageenan may be due to the irregular and hybrid structure of this polysaccharide. Only agar and lambda-carrageenan still undergo conformational transition at a high dose of 200 kGy. Gelation is observed for kappa-, iota-carrageenan up to a dose of 50 kGy while gelation is still observed at 100 kGy for agar. Increase in the hydrodynamic radius with decreasing temperatures for the non-irradiated carrageenans follows this order: lambda-carrageenan>kappa-carrageenan>iota-carrageenan. Slight increases in hydrodynamic radius were observed with irradiation.     

Physicochemical Properties of Biodegradable Polyvinyl Alcohol–Agar Films from the Red Algae <Emphasis Type="Italic">Hydropuntia cornea</Emphasis>

Agar obtained from the red alga Hydropuntia cornea was blended with polyvinyl alcohol (PVOH) in order to produce biodegradable films. In this study, we compare the properties of biopolymeric films formulated with agars extracted from H. cornea collected at different seasons (rainy and dry) in the Gulf of Mexico coast and PVOH as synthetic matrix. The films were prepared at different agar contents (0%, 25%, 50%, 75%, and 100%) and their optical, mechanical, thermal, and morphological properties analyzed. The tensile strength of PVOH–agar films increased when agar content was augmented. The formulation with 50% agar from rainy season (RS) had a significant higher tensile strength when compared to those from dry season (DS; p < 0.05). Tensile modulus also displayed an increasing trend and likewise, for 50% and 75% agar blends from RS showed higher values than those from DS (p < 0.05). In contrast, elongation at break decreased as the agar content increased, independently of the season. Environmental scanning electron microscopy images of PVOH–agar 75% biofilms from RS showed a homogeneous structure with good interfacial adhesion between the two components. The changes evidenced in the FTIR spectrum of this blend suggest that hydrogen bonding is taking place between the agar ether linkages (C-O-C) and the hydroxyl groups (OH) of the PVOH. Based on the above mentioned results, blends of PVOH and 75% agar from H. cornea collected in rainy season showed good properties for applications in the biodegradable packaging industry.     

Effects of seasons on the chemical structure and gel strength of Gracilaria pseudoverrucosa agar (Gracilariaceae, rhodophyta)

The seasonal effects on the chemical structure and rheological properties of Gracilaria pseudoverrucosa agar have been investigated using a sequential solvent extraction, ¹³C NMR and infrared spectroscopy, and gel strength measurements. The results showed that agar enriched in precursor to the agarobiose repeat unit were obtained from algae collected in summer. In contrast, algae collected in winter contained agar molecules richer in alkali-stable sulfate groups attributed in part to -galactose-4-sulfate. A similar total concentration of 6-O-methylated agarobiose repeat units was present in the agar from both algal samples but the distribution of the methylated disaccharide varied in the fractions. Agar fractions from the summer-collected sample had higher gel strength than those of the winter ones. Alkali treatment markedly improved the gel strength of the agar from the summer harvested seaweed. Different gel strengths were observed for the native and alkali-treated agar fractions extracted from the same algal sample and a gel strength comparable to that obtained for a commercial bacteriological grade agar was obtained from the alkali-treated 40% ethanol extract agar from the summer collected alga. The chemical and rheological variations due to seasonal changes are interpreted as reflecting the ratio of actively-growing (young) to resting (old) tissue in the alga and are proposed to represent a type of ‘secondarization’ of the algal cell-wall.     

The toxicity of potassium tellurite to Staphylococcus aureus in rabbit plasma fibrinogen agar

The potassium tellurite concentration, 0.01% w/v, in Baird-Parker agar has been recommended for plasma coagulase media such as pig or rabbit fibrinogen agar. Comparative tests have shown that with some strains of Staphylococcus aureus this level of potassium tellurite is too inhibitory and it should be reduced four-fold to 0.0025% w/v to maximize the isolation rate. It is postulated that egg yolk in Baird-Parker agar has a protective effect on staphylococci against the inhibitory action of tellurite.     

The toxicity of potassium tellurite to Staphylococcus aureus in rabbit plasma fibrinogen agar

The potassium tellurite concentration, 0.01% w/v, in Baird-Parker agar has been recommended for plasma coagulase media such as pig or rabbit fibrinogen agar. Comparative tests have shown that with some strains of Staphylococcus aureus this level of potassium tellurite is too inhibitory and it should be reduced four-fold to 0.0025% w/v to maximize the isolation rate. It is postulated that egg yolk in Baird-Parker agar has a protective effect on staphylococci against the inhibitory action of tellurite.     

Elongation of axons in an agar matrix that does not support cell locomotion

Axons elongate from 8-day embryonic chick spinal ganglion neurons cultured in a soft agar matrix or on a firm agar substratum. Glial cells from the same ganglia are unable to spread or show locomotory behavior (ruffling) in or on the agar, but do spread and show ruffling activity at the agarplastic interface or when replated from agar onto plastic. Axons elongated into or on agar retract much more slowly when treated with colchicine than do similar axons on plastic dishes.     

Isolation of an asporogenic (spoOA) protective antigen-producing strain of Bacillus anthracis.

We found that Congo red agar allows identification of sporulation-deficient Bacillus anthracis. Using Congo red agar, we isolated an asporogenic derivative of the protective antigen-producing strain B. anthracis delta Sterne-1(pPA102). Polymerase chain reaction and Southern hybridization analyses of DNA from the asporogenic mutant revealed that a deletion was present in spoOA, an essential gene for the initiation of sporulation. The deletion also encompassed the spoIVB homologue and a portion of the recN homologue. The avirulent spoOA strain delta Sterne-1(pPA102)CR4 is suitable for the safe production of protective antigen without endospore contamination of the vaccine production facility.     

The sclerotia of Coprinus lagopus

Summary Sclerotia of Coprinus lagopus were produced in the laboratory and observed microscopically from initiation to maturity. Isolated bulbous hyphal cells appeared below or on the agar surface as the primary sclerotial initials. Short interhyphal cells had expanded lateral walls and increased in number at their isolated locations, and were surrounded by a thick network of normal mycelium. Increasing in numbers, the bulbous cells developed a tightly compact colony devoid of agar if submerged below the agar surface and lacked normal hyphal filaments. The outer cells thickened formed cell lumina and developed as a protective rind to the mature sclerotium.     

Biosorption and elution of chromium from immobilized Bacillus coagulans biomass

Bacillus coagulans, a tannery wastewater isolate, previously shown to bind dissolved Cr(VI), retained its ability to biosorb Cr(VI) in different matrices. Polymeric materials like agar, agarose, calcium alginate and polyacrylamide were screened. Agarose emerged as the suitable candidate for biomass immobilization mainly due to its higher stability and integrity in acidic pH. Aptness of agarose as the matrix for B. coagulans biomass was revealed during Cr(VI) biosorption from natural wastewater.     

Improved soft-agar colony assay in a fluid processing apparatus

Summary The standard method for quantitating bone marrow precursor cells has been to count the number of colony-forming units that form in semisolid (0.3%) agar. Recently we adapted this assay for use in hardware, the Fluid Processing Apparatus, that is flown in standard payload lockers of the space shuttle. When mouse or rat macrophage colony-forming units were measured with this hardware in ground-based assays, we found significantly more colony growth than that seen in standard plate assays. The improved growth correlates with increased agar thickness but also appears to be due to properties inherent to the Fluid Processing Apparatus. This paper describes an improved method for determining bone marrow macrophage precursor numbers in semisolid agar.     

Sensitive Matrix Gel Diffusion Test for the Detection of Australia Antigen

A matrix gel diffusion (MGD) procedure with a sensitivity comparable to the complement fixation test (CF) has been developed for detecting Australia antigen in serum. The test utilizes a thin layer of agar (0.1 mm) with an applied plastic matrix. Reactants are introduced directly onto the surface of the agar through wells in the plastic matrix. End points obtained by CF with a panel of 11 sera varied from 1:8 to 1:512. When these sera were tested by MGD, end points for detection of antigen were within one dilution of that obtained by CF.     

Agars from three species of Gracilaria (Rhodophyta) from Yucatán Peninsula

Gracilaria cervicornis, Gracilaria blodgettii and Gracilaria crassissima growing along the coasts of Yucatán were investigated for their agar content. The effect of different concentrations of NaOH in the alkali treatment was evaluated. The three species of Gracilaria produced agars, both native and alkali treated, with different properties confirming the heterogeneity of the agar polymers in this genera. G. cervicornis produces agar polymers with an occurrence of methoxylation and sulphation at the C-6 of the beta-D-galactose residues, and with an extra methylation due to the presence of the 4-O-methyl-alpha-L-galactose residue. The presences of these residues is responsible for the extremely poor gelling ability of its agarocolloids, whose commercial value seems to be quite low. Agar extracts from G. blodgettii showed the typical pattern of unsubstituted agar with a very low degree of methylation on both galactose residues. The discrepancy found between sulphate content and NMR data of agar from this species requires a more detailed structural investigation. G. crassissima produces a good quality agar after sulphate precursor sequences have been removed by alkaline treatment, and it may be considered for exploitation as a source of commercial grade agar. Alkali treatment was effective both in removing alkali-labile sulphate and increasing the gel strength in G. crassissima but not in G. cervicornis and G. blodgettii.     

Stability of agar in the seaweed Gracilaria eucheumatoides (Gracilariales, Rhodophyta) during postharvest storage

The status of the cell-wall polysaccharide of the red seaweed, Gracilaria eucheumatoides upon postharvest storage was assessed in this study. The yield, chemical composition, physical and textural properties of alkali-treated agar extract was determined at different time intervals within 31 months of storage at dried state after harvest. Minimal fluctuation in agar yield was observed, ranging from 22.9% to 29.0%. The gel strength of agar extracts averaged 318gcm(-2) until the third month of storage but decreased considerably thereafter. The relative viscosity and molecular weight of the extracts varied inversely with storage time. Results indicated that both physical and textural parameters of agar generally decreased with storage time, likely due to depolymerization as indicated by decrease in molecular weight. Agar extracted from seaweeds up to 3 months of storage could be considered to exhibit gel quality suitable for food applications. Prolonged storage of the seaweed harvest is not recommended.     

Note: Microbial resistance of wool fabric treated with bis-Quats compounds

In this paper, the antibacterial activity against Bacillus pumilus, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa of the wool fabrics treated with new antimicrobial bis-quaternary surfactants, DABK and DABB, is studied. The activity was established on the basis of the agar diffusion and protective antibacterial test results and on the basis of scanning electron microscopy (SEM) observation. The results were compared with the HTAB, a monoquaternary surfactant of conventional use. The results from the agar diffusion and protective antibacterial tests do not enable us to confirm whether these compounds are potentially useful antimicrobial agents for the protection of textiles. However, SEM observations show clearly the efficacy of these compounds to protect the wool fabrics against the micro-organisms. SEM has been a useful technique for the assessment of antibacterial activity in textiles.