Development and growth of the lichen Rhizocarpon geographicum

The development of new areolae on the marginal hypothallus of the lichen Rhizocarpon geographicum (L.) DC was studied after complete or partial removal of the central areolae. New areolae developed slowly on the isolated hypothalli over two years. Development was similar when the areolae were completely removed and when the central areolae were separated from the marginal hypothallus by ‘moats’ 2 to 5 mm in width. However, in intact thalli, the marginal areolae developed rapidly during Jan. – June 1986 but showed periods of retreat from the margin during Oct. - Dec. 1985 and July – Sept. 1986. These results suggested that primary areolae may develop from free-living algal cells trapped by the hypothallus while secondary areolae may develop from zoospores produced by the thallus. Complete removal of the areolae resulted in no measurable radial growth of the marginal hypothallus over 18 months. Removal of the central areolae to within 1 and 2 mm of the hypothallus significantly reduced growth. These results suggest that the areolae may supply the hypothallus with carbon for growth. When the marginal hypothallus was experimentally removed a new hypothallus developed within one year. Regeneration occurred initially by retreat of the marginal areolae and later by new hyphal growth. The concentration of ribitol, arabitol and mannitol was measured in the areolae and marginal hypothallus on four occasions in 1985/6 in a population growing on a steep south facing rock surface. The three carbohydrates were present in significantly higher concentration in the areolae than in the hypothallus. Hence, the slow growth of this species may result from inhibited transport of carbohydrate from areolae to hypothallus.

Growth expectations of business owners:impact of human capital, firm characteristics and environmental transition

This paper presents an empirical study based on a survey of 399 owners of small and medium size companies in Lithuania. Applying bivariate and ordered probit estimators, we investigate why some business owners expect their firms to expand, while others do not. Our main findings provide evidence that SME owner's generic and specific human capital matter. Those with higher education and 'learning by doing' attributes, either through previous job experience or additional entrepreneurial experience, expect their businesses to expand. The expectations of growth are positively related to exporting and non-monotonically to enterprise size. In addition, we analyse the link between the perceptions of constraints to business activities and growth expectations and find that the factors, which are perceived as main business barriers, are not necessary those which are associated with reduced growth expectations. In particular, perceptions of both corruption and of inadequate tax systems seem to affect growth expectations the most.

Proteolytic activity amongst selected Saprolegnia species

Saprolegia ssp. effectively utilized the protein casein as a sole source of carbon, nitrogen and sulphur, indicating considerable proteolytic activity. In the presence of a more simple carbon source such as glucose, which was readily assimilated, catabolite repression was not observed and casein exploitation was enhanced. Free proteinase activity was not detected by a number of methods, irrespective of culture conditions. However, clearing by mycelia of skimmed milk agar or agar amended with bacteria demonstrated a close association between proteinases and hyphae, suggestive of natural immobilization of proteinases. Casein breakdown was accompanied by release of individual amino acids and ammonia. The latter, indicative of amino acid assimilation and metabolism, was also associated with an increase in pH of culture medium. Single amino acids did not support growth of Saprolegnia but in combination with other amino acids, methionine encouraged greatest biomass production. Certain groupings of amino acids affected growth in a manner which departed from that expected, as assessed by multifactorial analysis of variance, and either enhanced or reduced growth.

Entrepreneurs, expectations and business expansion:lessons from Lithuania

This article applies a multinomial logit estimator to investigate which factors affect SME owners' expectations to grow their businesses in Lithuania. Our findings provide evidence that SME owners' human capital (education) matters and that growth expectations are positively related to exporting. In addition, we analyse the link between the perceptions of business constraints and growth expectations and find that the factors, which are perceived as main business barriers, are not necessarily those which are associated with reduced growth expectations. However, perceptions of corruption seem to affect growth expectations the most.

Growth of experimentally reconstructed thalli of the lichen Parmelia conspersa

The centres of thalli of Parmelia conspersa (Ehrh. ex Ach.) Ach. were removed, the major lobes were separated from each other and from the substratum, and then the lobes were glued back together in their original configuration. The mean radial growth, the pattern of seasonal growth, and the degree of variation in growth between lobes of the reconstructed thalli, were similar to those of control thalli. When lobes were removed from thalli and glued apart from one another, the pattern of seasonal growth and the degree of variation in lobe growth were unaffected, but annual growth rates were reduced compared with lobes reconstructed into a thallus. Glueing the lobes together in a different configuration and constructing thalli in which each lobe came from a different ‘donor’ thallus did not influence the mean radial growth of the lobes or the degree of variation in lobe growth. These results suggest that although major lobes of P. conspersa are influenced by the proximity of their neighbours there is little chemical exchange between them. In addition, some thalli may form as a result of the fusion of lobes or propagules derived from different individuals. Copyright © 1984, Wiley Blackwell. All rights reserved

A nonantisense sequence-selective effect of a phosphorothioate oligodeoxynucleotide directed against the epidermal growth factor receptor in A431 cells

The overexpression of epidermal growth factor receptor (EGFr) has been implicated as a causative factor and a poor prognostic marker in a number of carcinomas. Therefore, strategies that down-regulate EGFr expression may be therapeutically useful. We designed antisense ODNs complementary to the initiation codon region of the EGFr mRNA and evaluated their efficacy in several tumor-derived cells, including the A431 cell line that express amplified levels of EGFr. A 15-mer phosphorothioate (PS) antisense ODN (erbB1AS15) induced a concentration-dependent reduction in proliferation that was accompanied by a change in the morphology of A431 cells into more tightly clustered and discrete colonies. A 15-mer sense (PS) control oligodeoxynucleotide (ODN) and a phosphodiester (PO) version of erbB1AS15 had little or no effect on cell number of morphology, and erbB1AS15 (PS) did not induce these effects in control cell lines expressing lower levels of EGFr. The effects of erbB1AS15 (PS) on A431 cells were not mediated by a true antisense mechanism in that there was no reduction in the level of EGFr mRNA or protein over a 24-hr period, as determined by Northern and Western blotting, respectively. However, autophosphorylation of the receptor was significantly reduced by erbB1AS15 (PS) and not by control ODNs. The results of further studies suggested that this effect was mediated by a direct, dose-dependent inhibition of the EGFr tyrosine kinase enzyme and was not due to impairment of either ligand-binding or receptor dimerization. These data suggest that erbB1AS15 (PS) can inhibit proliferation and alter the morphology of A431 cells by a sequence-selective, but nonantisense mechanism affecting receptor tyrosine kinase activity.

Experimental studies of lobe growth in the lichen Parmelia conspersa (Ehrh. Ex Ach.) Ach.

The radial growth of individual lobes of the foliose lichen, Parmelia conspersa (Ehrh. Ex Ach.) Ach. was studied to determine whether (1) adjacent lobes exchange carbohydrate and (2) marginal competition between lobes influences radial growth. In a survey of thalli of different size, the number of marginal lobes was linearly related to thallus circumference. However, the relationship between mean lobe width and thallus circumference was fitted by a second order polynomial. Hence, mean lobe width may reach a maximum in thalli approx. 3 cm in diameter. The interactions between marginal lobes were studied by either painting single lobes with acrylic paint or by removing lobes from the thallus. Painting the whole lobe virtually stopped its radial growth while partially painted lobes grew less than control lobes. The radial growth of a lobe was unaffected by either completely painting or removing its neighbour. Removal of both neighbouring lobes did not influence the radial growth of a lobe but severing the lobe from the thallus reduced its radial growth. In addition, lobe width increased significantly when both neighbouring lobes were removed. These results suggest that adjacent lobes have a considerable degree of independence and that there is little exchange of carbohydrate between them. In addition, marginal competition between adjacent lobes may restrict the lateral extension of the lobe and this may maintain a more constant mean lobe width in larger thalli. It is possible that the intensity of marginal competition between adjacent lobes may vary with thallus size and this could be a factor determining the growth curve of a foliose lichen throughout its life.

Matrix changes induced by transglutaminase 2 lead to inhibition of angiogenesis and tumor growth

Administration of active TG2 to two different in vitro angiogenesis assays resulted in the accumulation of a complex extracellular matrix (ECM) leading to the suppression of endothelial tube formation without causing cell death. Matrix accumulation was accompanied by a decreased rate of ECM turnover, with increased resistance to matrix metalloproteinase-1. Intratumor injection of TG2 into mice bearing CT26 colon carcinoma tumors demonstrated a reduction in tumor growth, and in some cases tumor regression. In TG2 knockout mice, tumor progression was increased and survival rate reduced compared to wild-type mice. In wild-type mice, an increased presence of TG2 was detectable in the host tissue around the tumor. Analysis of CT26 tumors injected with TG2 revealed fibrotic-like tissue containing increased collagen, TG2-mediated crosslink and reduced organized vasculature. TG2-mediated modulation of cell behavior via changes in the ECM may provide a new approach to solid tumor therapy.

Does radial growth of the lichen Parmelia conspersa depend exclusively on growth processes at the lobe tip?

The objective of this study was to test the hypothesis that the radial growth of lobes of the lichen Parmelia conspersa depends largely on growth processes which occur at the lobe tip. First, individual lobes were removed from thalli and portions of the lobe removed to within various distances from the tip. Radial growth of the lobe was unaffected until less than 2 mm of the lobe tip remained. Second, the surfaces of individual lobes were painted with acrylic paint leaving different portions of the lobe exposed. Painting lobes to within 0.5 mm and 1 mm of the tip substantially reduced radial growth. Third, the levels of ribitol, arabitol and mannitol were measured in different regions behind the lobe tip on four occasions during 1994. The concentration of the three carbohydrates was greatest at the lobe tip and the levels declined linearly with distance from the tip. Fourth, painting one vertical half of the lobe tip did not affect radial growth but artificially bisecting the lobe tip with a scalpel reduced radial growth. Although transport of carbohydrate from other regions of the lobe cannot be ruled out, the results support the hypothesis that radial growth in P. conspersa depends largely on processes within a region approximately 2 mm behind the lobe tip.

Experimental studies of hypothallus growth in the lichen Rhizocarpon geographicum

Removal of the areolae of the crustose lichen Rhizocarpon geographicum (L.)DC. resulted in either low or no measurable radial growth of the marginal hypothallus. Radial growth of the hypothallus was also significantly reduced compared with intact thalli when (1) areolae were removed to within 1 and 2 mm of the hypothallus and (2) a 5 mm wide ‘moat’ was created between the areolae and the hypothallus. Adding ribitol (0.01 M) to isolated hypothalli at 3-month intervals over 15 months results in total radial growth c. 60% that of intact thalli. Adding an arabitol/mannitol mixture (0.05 M arabitol, 0.03 M mannitol) increased radial growth compared with deionized water and ribitol treatments. Adding ribitol (0.7 M), arabitol (0.2 M) and mannitol (0.08 M) to the areolae of intact thalli had no significant effects on radial growth of the hypothallus. On a south-facing rock surface, isolated hypothalli grew at a similar rate to intact thalli for 2 months. Growth then declined and the hypothalli disappeared from the rock surface within 6 months. The effects of addition of carbohydrate suggest that the marginal hypothallus has the capacity to utilize exogeneous materials. However, in intact thalli in the field, the radial growth of the hypothallus is likely to be a result of transfer of materials from the areolae through hyphal connections.

The influence of bird droppings on the growth of lichen fragments transplanted to slate and cement substrates

The influence of bird droppings on the growth and fragmentation of five lichen species transplanted to slate and cement substrates was studied over a period of 15 months in South Gwynedd, Wales. The results suggested that at 15 months (1) thallus areas of Parmelia conspersa (Ehrh. Ex Ach.)Ach. were greater on both substrates with the addition of bird droppings with a greater increase on cement; (2) In Parmelia saxatilis (L.)Ach. And Parmelia glabratula ssp. fuliginosa (Fr. ex Duby)Laund., thallus areas were greatest on slate alone and least on cement with bird droppings; (3) in Physcia orbicularis (Neck.)Poetsch, thallus area was significantly reduced on cement alone compared with slate and cement treated with bird droppings; and (4) in Xanthoria parietina (L.)Th.Fr., thallus area was significantly greater on cement with bird droppings compared with slate and cement alone. These responses were attributable to the effect of the substrate and bird droppings on radial growth and the degree of fragmentation of the thalli. The results suggested that nutrient enrichment was more important than the substrate in determining the distribution of P. conspersa and Ph. orbicularis. However, the substrate and bird droppings were important in the remaining species, the data suggesting that P. saxatilis and P. glabratula ssp. fuliginosa would prefer nutrient-poor, siliceous rocks and X. parietina calcareous, nutrient enriched rocks in South Gwynedd.

Factors determining lobe growth in foliose lichen thalli

This study investigates the relative importance of climate, lobe morphology and lobe interactions in determining the radial growth of individual lobes in foliose lichen thalli. The radial growth of 75 lobes from thalli of Parmelia conspersa (Ehrh. Ex Ach.) Ach. and Parmelia glabratula ssp. fuliginosa (Fr. ex Duby) Laund. was measured over 22 successive months in relation to climatic factors. Individual lobes showed a fluctuating pattern of radial growth with alternating periods of fast and slow growth. In 17/75 (23%) of lobes studied, monthly radial growth was correlated with a climatic factor, usually total rainfall or the frequency of sunshine hours. In addition, the radial growth of 54 lobes of P. conspersa was measured over four months in relation to lobe morphology and the radial growth and morphology of adjacent lobes. Radial growth was correlated with lobe length and with the radial growth of adjacent lobes. In addition, the pattern of lobe branching appeared to be related to lobe width and to a lesser extent to lobe length and the width of adjacent lobes. The radial growth in one year of exceptionally long lobes which had grown beyond the thallus margin was similar to more normal lobes, but experimentally bisected lobes had significantly reduced radial growth compared with control lobes. These results suggested that the fluctuating pattern of radial growth in individual lobes may be determined by climate and the pattern of lobe branching. In addition, the pattern of lobe branching was related to lobe width and may be influenced by adjacent lobes.

The influence of bird droppings and uric acid on the radial growth of five species of saxicolous lichens

Bird droppings were applied over 1 year as a thick paste and as a suspension in deionized water to five species of lichens with different distributions on and off bird perching stones. The paste and suspension increased the radial growth of Parmelia conspersa while the paste increased the growth of Xanthoria parietina and reduced the growth and caused loss of colour in Parmelia glabratula ssp. fuliginosa. There were no statistically significant effects of paste or suspension on the growth of Physcia grisea or Parmelia saxatilis. In P. conspersa and X. parietina the growth responses were similar through the year but in P. glabratula the inhibitory effect of the paste was significant after 8 months growth. Application of a suspension of uric acid over 1 year had no statistically significant effects on the growth of P. conspersa, P. glabratula or X. parietina and was unlikely to be responsible for the effects of bird droppings on growth. The growth responses of the five species agreed well with their distributions in the field.

Factors affecting the growth of Cladophora in relation to river pollution

Nuisance growths of Cladophora have been associated with eutrophication. A review of the literature, however, reveals a scarcity of relevant experimental growth studies. Sampling experimental streams reveals that the addition of sewage effluent to good quality water alters the flora from that dominated by Potamogetan crispus to one dominated by CLadophora. Spatial and temporal differences in biomass of taxa present are discussed in the context of accompanying physicochemical data. In laboratory batch culture, growth of unialgal C. glomerata was accompanied by elevation of medium pH - considered largely responsible for the poor growth in such culture. However, appropriate experimental conditions and indices of growth were selected and the effects of various herbicides assessed. Diquat and terbutryne were shown to possess algicidal activity towards Cladophora. A closed continuous culture apparatus was developed: growth proceeded through lag, logarithmic and linear phases. Inoculum size and medium flow rate had significant effects on growth, and were standardized. In continuous culture, specific growth rate increased linearly with increased duration of light per day, up to 24 hours, and increased light intensity, up to 6000 lux - the highest intensity tested. Comparison of field and laboratory results suggests that ammonia toxicity is attributable to the undissociated form. In the laboratory, 185 µg/1 undissociated ammoniacal nitrogen reduced specific growth rate to 50% of that at 10 µg/1 undissociated ammcniacal nitrogen. 0.077-1.057 mg/1 NO2-N had no significant effect on growth. 7.2-15.2 mg/1 NO3-N had no significant effect on specific growth rate. Neither was any nitrate/phosphate interaction significant. At 4.9 mg/1 PO4-1, specific growth rate was only 48% of that at 1.9 g/1 P04-P. The critical medium PO4-P concentration was <0.1 mg/i. Specific growth rate was reduced to 50% of that in natural water by 0.036 mgCu/l, 0.070 mgzn/1 and 1.03 mgPb/l. Metal uptake was evaluated.

Studies on biofilm growth of Pseudomonas aeruginosa PAO1

The development of in vitro techniques to model the surface-associated mode of growth is a prerequisite to understanding more fully the physiological changes involved in such a growth strategy. Key factors believed to influence bacterial persistence in chronic infections are those of the biofilm mode of growth and slow growth rate. Methods for controlling Pseudomonas aeruginosa biofilm population growth rates were investigated in this project. This microorganism was incompatible with the in vitro 47mm diameter membrane filter-based biofilm technique developed for the study of Escherichia coli and Staphylococcus epidermidis by Gilbert et al (Appl. Environ. Microbiol. 1989, 55, 1308-1311). Two alternative methods were designed. The first comprised a 25mm diameter cellulose acetate membrane filter supported in an integral holder. This was found to be limited to the study of low microbial population densities with low flow rates. The second, based on a cylindrical cellulose fibre depth filter, permitted rapid flow rates to be studied and allowed growth rate control of biofilm and eluted cells. Model biofilms released cells to the perfusing medium as they grew and divided. The viability of released cells was reduced during, and shortly after, inclusion of ciprofloxacin in the perfusate. Outer membrane profiles of biofilm populations exhibited at least two bands not apparent in planktonic cells grown in batch and chemostat culture, and LPS profiles of biofilm populations showed variation with growth rate. Cell surface hydrophobicity of resuspended biofilm cells varied little with growth rate, whilst it decreased markedly for cells released from the biofilms as growth rate increased. Cells released from the biofilm were more hydrophilic than their sessile counterparts. Differing growth rates, LPS profiles and hydrophobicity are proposed to have a bearing on the release of cells from the adherent population.