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Biomedical subjects

R Marchant

Publications and source records attributed to R Marchant.

At least 37 records · Page 2Linked to original sources

Effect of environmental conditions on biological decolorization of textile dyestuff by C. versicolor.

Effects of environmental conditions such as pH, media composition, carbon and nitrogen sources, TOC/N ratio, and dyestuff concentrations on decolorization of reactive phytalocyanin type textile dyestuff Everzol Turquoise Blue G by white rot fungi, Coriolus versicolor 20) or low nitrogen concentration was essential for effective decolorization of the dyestuff. Dyestuff concentration should be lower than 500 mg/l for complete decolorization. Only partial decolorization was observed for dyestuff concentrations above 500 mg/l. Adsorption of the dyestuff on surfaces of the fungi was insignificant (<20%).

Journal Article↗

Decolorization of Remazol Black-B using a thermotolerant yeast, Kluyveromyces marxianus IMB3.

The ability of Kluyveromyces marxianus IMB3 to decolorize Remazol Black-B dye was investigated. The effect of environmental conditions, such as pH and temperature were examined. No noticeable effects on decolorization were observed when pH varied from 3.0-5.5. Maximum colour removal, 98%, was achieved at 37 degrees C. Little or no colour removal was detected when K. marxianus IMB3 was incubated under anaerobic conditions. Further investigation, in which decolorization was monitored under extreme temperatures and low pH (to inhibit growth) and using ten fold dense inoculum, revealed that decolorization was due to biosorption to the yeast cells and not due to a metabolic reaction.

Biomass↗

A drop-spreading technique to produce cytoplasm-free mitotic preparations from plants with small chromosomes.

A preparation technique has been developed for plants with small chromosomes, which produces large numbers of good-quality mitotic preparations. The technique employs a hydrochloric acid treatment to hydrolyse the cytoplasm, facilitating the subsequent removal of cytoplasmic debris. The evaporative force of a methanol-based fixative is exploited to disperse the cytoplasm and to deposit the chromosomes in a single optical plane. This technique permits detailed observations of chromosome morphology and karyotyping. The mitotic preparations are also suitable for the complex analysis associated with in-situ hybridization, as in studies of genome interaction in plant hybrids.

Chromosomes↗

Physiological and biochemical characterization of intergeneric hybrids of thermotolerant and non-thermotolerant yeasts.

Kluyveromyces-like intergeneric hybrids of thermotolerant Kluyveromyces marxianus and non-thermotolerant Saccharomyces cerevisiae, produced in a previous study by protoplasmic fusion, have been characterized. On molasses, these strains produced ethanol in excess of 6% (v/v) both at 30 and 45 degrees C as against 3% and 4.2% (v/v) by the former parent at 30 and 45 degrees C, respectively. In hybrids, the increase in ethanol production appeared to be a sequel to increased activities of alcohol dehydrogenase and pyruvate kinase, derived probably from S. cerevisiae parent. Hybrid ADH-isozyme pattern on polyacrylamide gel corroborated the presence of S. cerevisiae ADH in the tested hybrids. Regression analyses indicated a positive correlation between ethanol production and ADH or PK or both (r approximately 0.76-0.84).

Hybridization, Genetic↗

Preservation of viable biological samples for experiments in space laboratories.

Standard viable preservation methods for biological samples using low temperatures have been investigated concerning their storage capabilities under higher temperature levels than usual. For a representative set of organism classes (plants, mammalian cells, arthropods and aquatic invertebrates), the minimum appropriate storage conditions have been identified by screening storage temperatures at -196 degrees, -80 degrees, -20 degrees, +4 degrees, +20 degrees/25 degrees C for periods from 2 days to 4 weeks. For storage below 0 degree C, as a typical cryopreservative, dimethylsulfoxide (DMSO) was used. For some samples, the addition of trehalose (as cryopreservative) and the use of a nitrogen atmosphere were investigated. After storage, the material was tested for vitality. The findings demonstrated that acceptable preservation can be achieved under higher storage temperatures than are typically applied. Small, dense cultured plant cells survive for 21 d when moderately cooled (+4 degrees to -20 degrees C); addition of trehalose enhances viability at -20 degrees C. For mammalian cells, the results show that human lymphocytes can be preserved for 3 d at 25 degrees C, 7 d at 4 degrees C and 28 d at -80 degrees C. Friend leukaemia virus transformed cells can be stored for 3 d at 25 degrees C, 14 d at 4 degrees C and 28 d at -80 degrees C. Hybridoma cells can be kept 7 d at 4 degrees C and 28 d at -20 degrees C or -80 degrees C. Model arthropod systems are well preserved for 2 weeks if maintained at lower temperatures that vary depending on the species and/or stage of development; e.g., 12 degrees C for Drosophila imagoes and 4-6 degrees C for Artemia nauplii. For aquatic invertebrates such as sea urchins, embryonic and larval stages can be preserved for several weeks at +6 degrees C, whereas sperm and eggs can best be stored at + 4 degrees C for up to 5 d at maximum. These results enhance the range of feasible space experiments with biological systems. Moreover, for typical terrestrial preservation methods, considerable modification potential is identified.

Animals↗

Fermentation of molasses using a thermotolerant yeast, Kluyveromyces marxianus IMB3: simplex optimisation of media supplements.

The use of molasses as a substrate for ethanol production by the thermotolerant yeast Kluyveromyces marxianus var. marxianus was investigated at 45 degrees C. A maximum ethanol concentration of 7.4% (v/v) was produced from unsupplemented molasses at a concentration of 23% (v/v). The effect on ethanol production of increasing the sucrose concentration in 23% (v/v) molasses was determined. Increased sucrose concentration had a similar detrimental effect on the final ethanol produced as the increase in molasses concentration. This indicated that the effect may be due to increased osmotic activity as opposed to other components in the molasses. The optimum concentration of the supplements nitrogen, magnesium, potassium and fatty acid for maximum ethanol production rate was determined using the Nelder and Mead (Computer J 7:308-313, 1965) simplex optimisation method. The optimum concentration of the supplements were 0.576 g1(-1) magnesium sulphate, 0.288 g1(-1) potassium dihydrogen phosphate and 0.36% (v/v) linseed oil. Added nitrogen in the form of ammonium sulphate did not affect the ethanol production rate.

Culture Media↗

Biological treatment of distillery waste for pollution-remediation.

The biological treatment of spent wash from molasses distilleries was investigated. Analysis of raw spent wash showed it to be a recalcitrant waste, with a high COD of 85,170 mg/l and containing inhibitory phenolic compounds. Reverse phase thin layer chromatography identified gallic and vanillic acid present in spent wash. The fungi Geotrichum candidum, Coriolus versicolor, Phanerochaete chrysosporium and Mycelia sterilia were screened for their ability to decolourize spent wash and to reduce the COD level. A 10 day pretreatment with Geotrichum candidum at 30 degrees C resulted in reducing the COD by 53.17% and total phenols by 47.82%, enabling other bioremediating organisms to grow. Coriolus versicolor immobilized in a packed-bed reactor reduced the COD of spent wash by a further 50.3%, giving an overall reduction in COD of 77% to 15,780 mg/l. A small amount of decolourization was achieved (4.2%), although the spent wash was still coloured. Present studies are encouraging and indicate that it is possible to bioremediate spent wash using a multi-stage treatment process involving an initial pretreatment step with Geotrichum candidum.

Bacteria↗

Factors affecting the genetic engineering of plants by microprojectile bombardment.

Since its development in the mid-1980s, microprojectile bombardment has been widely employed as a method for direct gene transfer into a wide range of plants, including the previously difficult-to-transform monocotyledonous species. Although the numerous instruments available for microprojectile-mediated gene delivery and their applications have been widely discussed, less attention has been paid to the critical factors which affect the efficiency of this method of gene delivery. In this review we do not wish to describe the array of devices used for microprojectile delivery or their uses which have already been definitively described, but instead wish to report on research developments investigating the factors which affect microprojectile-mediated transformation of plants.

Journal Article↗

Production, partial characterization, and potential diagnostic use of salicylate hydroxylase from Pseudomonas putida UUC-1.

An unusual strain of Pseudomonas putida UUC-1 capable of growth at high salicylate concentration (10 gl-1) was investigated with the aim of developing an assay and a biosensor system for determining salicylate in body fluids by utilizing the salicylate hydroxylase enzyme. Medium and growth condition optimization were carried out under chemostat conditions. The highest biomass yield was at 4.0 gl-1 salicylate, 25 degrees C, pH 6.5, and 0.2 h-1 dilution rate. Growth occurred at up to 0.45 h-1 dilution rate, producing 236 Ul-1 enzyme activity and an output of 424 U h-1. The activity and productivity were higher than any reported in the literature for this enzyme. It had a Km value of 2.07 +/- 0.32 microM and an M(r) of approximately 43,000. In addition, its specific activity in the crude extract (0.8-0.9 U mg-1 protein) was similar to the commercially available enzyme. No plasmid DNA was detected in this strain, and no salicylate-negative isolates were obtained when curing with mitomycin C. It is therefore proposed that our strain has a chromosomally located inducible salicylate hydroxylase gene that enables it to grow at high salicylate. This strain also offers a means of cheaply producing large quantities of the enzyme through standard fermentation techniques.

Biosensing Techniques↗

Production of the enzyme dihydrofolate reductase by methotrexate-resistant bacteria isolated from soil.

Six bacterial cultures isolated from soil were capable of growing in the presence of methotrexate (MTX). Two strains, PFR-1 and 3, developed resistance to 500 micrograms cm-3 MTX in the medium and produced elevated levels of the enzyme dihydrofolate reductase (EC 1.5.1.3): 2580 and 2702 U dm-3 compared to the sensitive parent strains (28 and 35 U dm-3). Isolate PFR-3 showed maximum enzyme production (4950 U dm-3, specific activity 12.56 U mg-1 in flasks and 5737 U dm-3, specific activity 14.80 U mg-1 in 5-dm3 fermenter) during exponential phase of growth (6 h) at 37 degrees C and pH 7.0.

Chromatography, High Pressure Liquid↗

Dihydrofolate reductase synthesis in continuous culture using a methotrexate-resistant Escherichia coli.

A methotrexate-resistant strain of Escherichia coli Type I produced exceptionally high levels of the enzyme dihydrofolate reductase (EC 1.5.1.3) in 6 h of batch fermentation. The culture had markedly improved performance under chemostat culture conditions in terms of enzyme yield and output. Temperature, pH, dilution rates, and nutrient composition were optimized under chemostat culture conditions. A maximum enzyme yield of 10,360 U l-1 and a specific activity of 22.84 U mg-1 were obtained under chemostat conditions at pH 7.0, temperature 37 degrees C, and dilution rate 0.2 h-1, using media containing 1.0 and 0.6% (w/v) dextrose and yeast extract, respectively. The culture's performance and enzyme yields in chemostat and the feasibility of large-scale production are discussed.

Bacteriological Techniques↗

In vivo biocompatibility studies. I. The cage implant system and a biodegradable hydrogel.

A cage implant system has been utilized to examine the in vivo biocompatibility of a biodegradable hydrogel, poly(2-hydroxy-ethyl-L-glutamine) (PHEG). This system permits the quantitative determination of the components of the inflammatory exudate which surrounds the implanted polymer within the cage system. This system permits the serial examination of exudate components without sacrificing the animal. In addition, this system allows the subsequent removal of the polymer for surface and mechanical studies. Following implantation of the biodegradable hydrogel, quantitative and differential white cell counts of the exudates were determined over a 21-day period. In addition, concomitant extracellular enzyme analyses for alkaline phosphatase, acid phosphatase, prostatic acid phosphatase, leucine amino-peptidase, and Cathepsin B1 were determined. Corresponding control samples from exudates of the cage implant without the polymer were also determined. The two-tailed Student's t-test for unpaired samples was used to statistically compare the control and implanted polymer values for these respective analyses at the various time periods. A comparison of the cellular response for the control system and the PHEG system did not show statistically significant differences during the first 7 days following implantation. The acute inflammatory response, polymorphonuclear leukocyte predominant, was followed by a mild chronic inflammatory response, macrophage and lymphocyte predominant, and during this time period, 8-14 days, macrophages were present in significantly larger numbers for the PHEG system when compared to the control values. Enzymic analysis of the exudates revealed statistically significant differences between control and PHEG values at time intervals where no differences were noted in cell density or population. These results are discussed in terms of cell-polymer interactions leading to cellular activation and enhanced enzyme exocytosis by the inflammatory cells. Stress-strain measurements on implanted PHEG samples showed that significant in vivo degradation had occurred during the acute inflammatory phase of the response, i.e., the first 7 days.

Acid Phosphatase↗

Neonatal lamb losses due to feral pig predation.

An investigation into causes of low lamb marking percentages was made on a property in north-west New South Wales from 1971 to 1975. Investigations revealed that from 11 to 70% of the ewes in lamb were losing all their lambs. Observations suggested that feral pig predation was a factor in the perinatal loss. In 1975, 2 groups of ewes were placed in adjoining paddocks prior to lambing. Feral pigs were excluded from one paddock for most of lambing by means of an electric fence. In this paddock, 117% of lambs were marked compared with 80% in the adjoining paddock. It was estimated that in 1975 over 600 lambs were killed by feral pigs from 1,422 ewes lambing in paddocks with feral pigs. The problems involved in the diagnosis of feral pig predation are discussed.

Animal Population Groups↗

Ultrastructure of an indigotin-producing dome mutant of Schizophyllum commune.

Electron microscopic observations of an indigotin-producing dome mutant of Schizophyllum commune Fr. have shown that large wall ingrowths occur within the hyphae. These ingrowths are coupled with morphological abnormalities produced by the dome mutation. The pigment indigotin appears to be produced by progressive condensation within vacuoles and to a lesser extent within the wall ingrowths. Cytochemical techniques have shown that the wall ingrowths are similar in structure to the hyphal walls. there was no evidence for the passage of condensed indigotin into the medium; the pigment granules found in the medium must therefore form outside the hyphae.

Agaricales↗

The effect of cytochalasin B on hyphal morphogenesis in Polyporus biennis.

Cytochalasin B inhibited the radial growth rate of Polyporus biennis, and caused an increase in hyphal density through a reduction in the distance between successive branches. Cytochalasin B also produced irregular hyphal profiles and, in a small percentage of hyphae, forked apices. The position of clamp connexions was little affected by cytochalasin B, but the developmental process was specifically inhibited during initiation and during the last two stages, when contact and dissolution of the clamp were occurring. There were no major disruptions of the ultrastructure of the dolipore/parenthesome septum caused by cytochalasin B treatment.

Basidiomycota↗