Production Of Extracellular Material Research Articles

Lichen 1. Solo fungal and algal partners

Lichens are slow-growing niche-constructing organisms that form a thallus via scripted symbiotic/mutualistic relationships between fungi, algae, and bacteria, and that are distributed across nearly all terrestrial ecosystems. Here we use quick-freeze deep-etch electron microscopy (QFDEEM) to probe the ultrastructure of a lichen-forming fungus, Cladonia grayi, and its partner alga, Asterochloris glomerata, grown separately (solo) in the laboratory. The solo fungus resembles its lichenized counterpart in: a) general cellular organization; b) the capacity to form lateral branches; c) the production of extracellular materials; and d) the generation of acellular hyphae that we call struts. It differs in carrying a fibrillar coat on its wall exterior and in often adopting two novel plasma-membrane configurations called pleated and pitted. The solo alga also resembles its lichenized counterpart in general cellular organization, but its algaenan-based wall carries a fibrillar coat not evident in the lichen, and its pyrenoid and plastoglobule endowments are less well developed. These findings are followed by four reports on the QFDEEM ultrastructure of four species of lichens, where we describe the differentiations that occur when the fungi and algae are living together in community with bacteria.

Diffusive public goods and coexistence of cooperators and cheaters on a 1D lattice.

Many populations of cells cooperate through the production of extracellular materials. These materials (enzymes, siderophores) spread by diffusion and can be applied by both the cooperator and cheater (non-producer) cells. In this paper the problem of coexistence of cooperator and cheater cells is studied on a 1D lattice where cooperator cells produce a diffusive material which is beneficial to the individuals according to the local concentration of this public good. The reproduction success of a cell increases linearly with the benefit in the first model version and increases non-linearly (saturates) in the second version. Two types of update rules are considered; either the cooperative cell stops producing material before death (death-production-birth, DpB) or it produces the common material before it is selected to die (production-death-birth, pDB). The empty space is occupied by its neighbors according to their replication rates. By using analytical and numerical methods I have shown that coexistence of the cooperator and cheater cells is possible although atypical in the linear version of this 1D model if either DpB or pDB update rule is assumed. While coexistence is impossible in the non-linear model with pDB update rule, it is one of the typical behaviors in case of the non-linear model with DpB update rule.

A two-compartment mechanochemical model of the roles of transforming growth factor [formula omitted] and tissue tension in dermal wound healing

The repair of dermal tissue is a complex process of interconnected phenomena, where cellular, chemical and mechanical aspects all play a role, both in an autocrine and in a paracrine fashion. Recent experimental results have shown that transforming growth factor ‐ β (TGF β ) and tissue mechanics play roles in regulating cell proliferation, differentiation and the production of extracellular materials. We have developed a 1D mathematical model that considers the interaction between the cellular, chemical and mechanical phenomena, allowing the combination of TGF β and tissue stress to inform the activation of fibroblasts to myofibroblasts. Additionally, our model incorporates the observed feature of residual stress by considering the changing zero-stress state in the formulation for effective strain. Using this model, we predict that the continued presence of TGF β in dermal wounds will produce contractures due to the persistence of myofibroblasts; in contrast, early elimination of TGF β significantly reduces the myofibroblast numbers resulting in an increase in wound size. Similar results were obtained by varying the rate at which fibroblasts differentiate to myofibroblasts and by changing the myofibroblast apoptotic rate. Taken together, the implication is that elevated levels of myofibroblasts is the key factor behind wounds healing with excessive contraction, suggesting that clinical strategies which aim to reduce the myofibroblast density may reduce the appearance of contractures.

Correlation between the presence of Gonyaulax fragilis (Dinophyceae) and the mucilage phenomena of the Emilia-Romagna coast (northern Adriatic Sea)

The formation of massive amounts of suspended mucilaginous organic matter which periodically affects the Adriatic Sea, has been regarded as a complex physico-chemical phenomenon resulting from the production of extracellular material by phytoplankton. Although the exact cause has remained obscure, the mechanism of its formation has usually been considered to be a long-term process, starting after the late winter-early spring blooms, and involving the participation of various algal species, mainly within the diatom group. In this paper we report on the results of a phytoplankton monitoring programme in northern Adriatic seawaters off the Emilia-Romagna coast of Italy which revealed the constant concomitant presence of the dinoflagellate Gonyaulax fragilis (Schütt) Kofoid and mucilaginous formations. In the early stages of the phenomenon the dinoflagellate was clearly observable by microscopic examination in the mucilage, but as the mucilage aged this alga almost completely decomposed and diatom cells increased in number and became predominant. Although characterized by a slow growth rate in culture, in natural seawater G. fragilis was observed to reach cell densities of up to 7.0×10 6 cells l −1. The results of this study lead us to propose the hypothesis that the appearance of mucilage in the water column of the Adriatic Sea is the consequence of a seasonal growth of this dinoflagellate favoured by specific environmental circumstances.

The surface of Erysiphe graminis and the production of extracellular material at the fungus – host interface during germling and colony development

Low temperature scanning electron microscopy was used to study the development of Erysiphe graminis DC f.sp. avenae Marchal from germination through infection to sporulation of the fungal colony. To clarify resolution of the fungus–host interface and facilitate interpretation of fungal surface structures, epicuticular waxes were removed from host leaves before inoculation. Whereas conidia were covered in spinelike protrusions or globular bodies, young germ tubes, appressoria, and hyphae were initially smooth walled, but by 15 h after inoculation, wartlike bodies, resembling globular bodies on conidia, were present on first appressorial lobes; these increased in number, eventually covering the appressorium surface and appearing on contiguous hyphae. Wartlike bodies also appeared at junctions of hyphal branches, on hyphal appressoria, and on conidiophore basal cells. Their function, if any, is unknown. The meristematic zone, at the apex of the conidiophore basal cell, remained smooth walled, but globular bodies appeared on the wall of young conidia as soon as the limiting septum had formed. Observations with the fungus in situ revealed the presence of amorphous extracellular material around primary germ tubes and appressorial lobes. Extracellular material was also present beneath appressorial germ tubes and hyphae but it was hidden unless the fungus was displaced. It could not be seen beneath conidia. The extracellular material appeared to be adhesive, sticking the fungus firmly to the host surface. Removal of the fungus showed that the extracellular material was deposited close to the tip of developing germ tubes and hyphae. It was particularly thick around primary germ tubes and appressorial lobes, and a discrete ring of extracellular material was often visible around penetration pores (holes in the leaf surface seen beneath primary germ tubes and appressorial lobes). In addition to its adhesive properties, the extracellular material may act as a matrix in which fungal enzymes are sited and focused for attack on the host. Key words: Erysiphe graminis, low temperature scanning electron microscopy, extracellular material, fungal adhesion, fungal surface morphology.

Biofilm Development and Sanitizer Inactivation of Listeria monocytogenes and Salmonella typhimurium on Stainless Steel and Buna-n Rubber

Biofilm formation by seven strains of Listeria monocytogenes and one strain of Salmonella typhimurium on stainless steel and Buna-n rubber was examined under two nutrient conditions. The type of surface, nutrient level, and organism influenced biofilm development and production of extracellular materials. Buna-n had a strong bacteriostatic effect on L. monocytogenes, and biofilm formation on Buna-n under low nutrient conditions was reduced for four of the seven strains tested. Buna-n was less bacteriostatic toward S. typhimurium. It inhibited the growth of several other pathogens to varying degrees. An ethylene propylene diamine monomer rubber was less inhibitory than Buna-n, and Viton rubber had no effect. The effectiveness of sanitizers on biofilm bacteria was examined. Biofilms were challenged with four types of detergent and nondetergent sanitizers. Resistance to sanitizers was strongly influenced by the type of surface. Bacterial biofilm populations on stainless steel were reduced 3–5 log by all the sanitizers, but those on Buna-n were resistant to these sanitizers and were reduced less than 1–2 log. In contrast, planktonic (suspended) bacteria were reduced 7–8 log by these sanitizers. Chlorine and anionic acid sanitizers generally removed extracellular materials from biofilms better than iodine and quaternary ammonium detergent sanitizers. Scanning electron microscopy demonstrated that biofilm cells and extracellular matrices could remain on sanitized biofilm cells and extracellular matrices could remain surfaces from which no viable cells were recovered.

Growth inhibition of cartilaginous embryonic chick tibiae by ionizing radiation: an in vitro study

The radiation effect on the longitudinal growth of cartilaginous tibiae of 6,5 day old chick embryos is studied in vitro. After irradiation length is measured daily during the 3 days culture period and incorporation of (3)H-thymidine, (3)H-uridine or (3)H-proline is sampled by liquid scintillation counting. Selective tibiae are videorecorded during the incubation period. Morphology is studied by light microscopy, autoradiography and electron microscopy.Growth inhibition is only restricted to 5% at 20 Gy up to 20% at 150 Gy. The (3)H-uridine and (3)H-proline incorporation is only decreased by 21 and 32% at 150 Gy. (3)H-thymidine incorporation on the contrary shows 17% decrease at 2 Gy. Irradiation at 200 Gy gives a total growth arrest and stops incorporation of all used labeled compounds in 75% of the specimens. L.M. and E.M. show no morphological differences between irradiated and sham irradiated tibiae until 150 Gy. At 200 Gy however necrosis of the cells is observed.We conclude that longitudinal growth of cartilaginous tibiae in organ culture is not correlated with DNA synthesis but with cell hypertrophy and production of extracellular material. RNA transcription and synthesis of collagen fibers are radioresistant processes. This explains the limited influence of irradiation on the longitudinal growth.

Scanning electron microscopy of dairy equipment surfaces contaminated by two milk-borne micro-organisms.

Ethanol dehydration followed by argon replacement induced drying (ARID) was found to be a suitable method for the preparation of glass, stainless steel and rubber surfaces which had been in contact with inoculated milk and which were to be examined using scanning electron microscopy (SEM). This technique was used to examine samples of all three materials which had been subjected to both single and repeated inoculation with whole milk containing a Pseudomonas sp. or a Micrococcus sp. and incubated for various periods. Some samples were also prepared for SEM using a cryofixation technique. The Pseudomonas sp. was found to proliferate on glass and stainless steel surfaces but not on rubber. Due to the clumping tendency of the Micrococcus sp. proliferation of this organism was more difficult to assess accurately. In general there was no difference in results obtained between single and repeated inoculation. Various factors which may have aided attachment of micro-organisms to surfaces were identified viz., surface channels present in stainless steel, milk deposits and the production of extracellular material. The value of using both the cryofixation and chemical preparatory techniques for the identification of artifacts is discussed.

Efficacité de huit désinfectants sur trois types de surfaces contaminées par Pseudomonas aeruginosa

The disinfecting capacity of eight commercial chemical products was evaluated by the use--dilution method given by the Associated of Official Analytical Chemists (AOAC) on three types of surface material (steel, aluminum, and plastic). For most products tested the limit concentration was 10 times higher for disinfecting aluminum and plastic surfaces than stainless steel. As observed on the scanning electron microscope, the number of bacteria deposited on the surface and the production of extracellular material on polypropylene by Pseudomonas aeruginosa ATCC 15442 would explain the observed differences. The applicability of the AOAC method or other techniques for the evaluation of the disinfecting capacity on different surfaces is discussed.

Long-term culture of human aortas. Development of atherosclerotic-like plaques in serum-supplemented medium.

Segments of human thoracic aorta were maintained in long-term explant culture for 18 weeks in serum-supplemented medium. The aortas were grossly normal in appearance, and random samples fixed for light microscopy prior to culture revealed a normal morphology. The intima contained no more than five layers of smooth muscle cells. After 7 days in culture, the intima was noticeably thicker than the uncultured segments. The increased thickness was due to proliferating smooth muscle cells and production of extracellular material. After several months in culture, extracellular material consisting of collagen and flocculent material was present in areas resembling atherosclerotic fibrous plaques. A peripheral growth, which formed around the explant, was composed of fibroblastlike cells and added to the overall thickness of the intima. However, aortic segment maintained for up to 2 months in serum-free culture medium showed no cellular proliferation. This study demonstrates that changes resembling early stages of atherosclerosis occur in human aortas maintained in explant culture using routine culture procedures.

Production of Extracellular Material by Streptococci Associated with Subacute Bacterial Endocarditis

Production of Extracellular Material by Streptococci Associated with Subacute Bacterial Endocarditis