Preparation of enriched fractions from cerebral cortex containing isolated, metabolically active neuronal and glial cells

1. A procedure has been developed for the separation of intact metabolically active neuronal and glial cells in bulk from rat cerebral cortex. Separation depended on dispersion of the tissue in a Ficoll medium followed by centrifugation on a discontinuous Ficoll gradient. Up to 1.5x10(7) neuronal cells could be collected from 12 brains within 3hr. The morphological appearance of these cells seemed good, and the fraction was 8.5-fold purified in terms of dry weight. Average dry weight per neuron was 2300mumug. Maximum glial contamination of the neuronal fraction was 11% as determined by carbonic anhydrase measurements. The glial fraction was free from neurons but contained various subcellular contaminants. 2. Concentrations of nucleic acids, phospholipid, protein and phosphoprotein were determined in the separated fractions. The neuronal fraction was richer than the glial in all except phospholipid. Succinate dehydrogenase was equally distributed between neurons and glia but the neuronal fraction was 1.8-fold enriched in cytochrome oxidase. 3. Measurement of respiration by the cells showed an endogenous uptake of 117mmumoles of oxygen/mg./hr. in neurons, and 173mmumoles of oxygen/mg./hr. in glia. Addition of substrate at 10mm stimulated uptake to similar values in both fractions. With glucose it was 390, with pyruvate 355, and with glutamate 215mmumoles of oxygen/mg./hr. This represented a larger stimulation of neuronal than of glial respiration compared with the basal level. 4. Respiration in cell suspensions was 70-80% of that of slices, whereas fractionated tissue homogenates had respiratory rates of only one-third those of the cell suspensions. Lactate dehydrogenase content of cell suspensions was maintained during gradient centrifugation and washing. 5. The possible uses of isolated cell preparations are discussed.     

Submergence(Fermentation)Culture of Anisodus acutanguhts Cells and Identification of Hyoscyamine and Scopolamine in the Cultured Ceils

The growth rate of Anisodus acutangulus cells in the submergence culture was 1.5 g dry wt/l/day, a rate 3 times as that in the suspension culture and more than 10 times over that of the solid static culture: However, the contents of hyoscyamine (0.203 mg/g dry wt) and scopolamine (0.178 mg/g dry. wt) in submergence culture were only slightly higher than those in the two other cultures. But when the 12-day-old submergence culture was supplemented with phenylalanine (5m mol/l) and kinetin (0.1mg/l), it was observed that not only the cell growth rate was increased but also the cellular content of hyoscyamine was raised to a level of 0.217 mg/g dry wt., and that scopolamine to 0.412 mg/g dry wt. The contents of these two alkaloids represented 1.1 and 2.3 times respectively the value of the culture without phenylalanine and kinetin supplements. The optimum date for harvesting the A. acutangulus culture Was on the 14th day of the culture. The monomers of hyoscyamine and scopolamine isolated from the cultured cells were purified and recrystalized, and then identifited as the two compounds in question by the thin-layer chromatography, melting point determination, and ultraviolet, infra-red and nuclear magnetic resonance spectra. In this paper, we also summarize and discuss the results of A. acutangulus culture experiments performed in the past 8 years. Our finding seems to inclieate that following a pilot production trial, the tissue culture method could well be employed to produce hyoscyamine and scopolamine from A. acutangulus cells on an industrial scale.     

Continuous Cultivation of Glucose-Limited Bean Cells (Phaseolus vulgaris L.) in a Modified Bacterial Fermentor

A commercially available bacterial fermentor was modified toallow the continuous cultivation of glucose-limited bean cells(Phaseolus vulgaris L.) at a defined pH. A steady state wasreached using a dilution rate of 0?004 h^–1 and was maintainedfor 4 months; during this period the dry weight and packed cellvolume of the culture remained constant; the yield for glucosewas 0?34 (dry wt./dry wt) and for carbon 0?27 (g atom/g atom).Glucose-limited cells had lower C/N ratios both for cytoplasmand cell walls than cells in the exponential phase of growthin batch cultures.     

Metabolic control mechanisms in mammalian systems. Hormonal regulation of phosphofructokinase in the rat prostate and seminal vesicles

1. The hormonal regulation of phosphofructokinase was investigated in the accessory reproductive organs of the orchidectomized rat. 2. Phosphofructokinase activities declined to 51% and 47% in the prostate and 9% and 6% of the normal values in seminal vesicles 4 and 8 weeks after castration respectively. Administration of testosterone (100mug./100g. body wt.) for 3 days reversed substantially the effects of orchidectomy, and phosphofructokinase activity increased to 173% in the prostate and 536% in seminal vesicles as compared with the values of castrated controls. 3. Time-course studies demonstrated that after a single injection of testosterone (5mg./100g. body wt.) phosphofructokinase activity was maximally elevated to 236% in the prostate and 342% in seminal vesicles at 24hr. 4. Dose-response studies revealed that 2.5mg. of testosterone propionate/100g. body wt. was the minimal amount necessary to induce significant increases in enzyme activity in both accessory sex organs; maximal increases were obtained with a dose of 5mg./100g. body wt. 5. The observed enzyme increases induced by testosterone were inhibited by the simultaneous administration of oestradiol-17beta, and phosphofructokinase activity in this group of rats remained at 97% in the prostate and 137% of the control values in seminal vesicles. Oestradiol-17beta by itself failed to produce any significant effect on enzyme activity in either of these secondary sexual tissues. 6. The nature of the testosterone-induced increases in phosphofructokinase activity was studied by using a variety of inhibitors of RNA and protein synthesis. Cycloheximide, 5-fluorouracil and ethionine largely blocked the androgen-stimulated rise in enzyme activity observed 24hr. after steroid injection. The inhibitory effect of ethionine was completely reversed by the simultaneous administration of methionine. 7. Actinomycin, which is known to inhibit the synthesis of messenger RNA as well as the synthesis of other cellular RNA fractions, when given simultaneously with the hormone, also inhibited the testosterone-induced increases in prostatic and seminal-vesicular phosphofructokinase. However, when the antibiotic was given 6 or 12hr. after injection of the steroid, practically no inhibition of phosphofructokinase induction was obtained. This indicates that, once the enzyme-forming machinery is turned on and allowed to operate for a few hours, actinomycin is incapable of reversing the hormone-induced enzyme responses. 8. The results presented suggest that new RNA and protein synthesis may be involved in the observed androgen-induced increases in phosphofructokinase activity in the prostate and seminal vesicles of the orchidectomized rat.     

Construction of a high-biomass, iron-enriched yeast strain and study on distribution of iron in the cells of Saccharomyces cerevisiae

A high-biomass, iron-enriched Saccharomyces cerevisiae ZYF-15 was constructed by interspecies protoplast fusion. Under optimal fermentation condition, the biomass and iron content of the strain reached 11 g l(-1) and 25 mg Fe g(-1) dry cells, respectively. About 96% of enriched iron is converted into organic iron, which is mainly in cell walls and vacuoles with some bound to DNA, RNA and protein.     

Combined effect of ascorbic acid and selenium supplementation on alcohol-induced oxidative stress in guinea pigs

Oxidative stress is a key step in the pathogenesis of ethanol associated liver injury. Ethanol administration induces an increase in lipid peroxidation either by enhancing the production of oxygen reactive species or by decreasing the level of endogenous antioxidants. In this present study, four groups of male guinea pigs (Cavia porcellus) were maintained for 45 days as follows: Control group (1 mg ascorbic acid (AA)/100 g body wt./day); Ethanol group (1 mg AA/100 g body wt./day+900 mg ethanol/100 g body wt./day); Selenium+AA group (25 mg AA+0.05 mg sodium selenite/100 g body wt./day); Ethanol+Se+AA group (25 mg AA+0.05 mg sodium selenite/100 g body wt.+900 mg ethanol/100 g body wt./day). Malondialehyde (MDA), hydroperoxides (HP) and conjugated dienes (CD) were significantly increased, while the activities of scavenging enzymes superoxide dismutase (SOD) and catalase were reduced in the alcohol administered groups. Co-administration of Se+AA along with alcohol increased the activities of scavenging enzymes and reduced the lipid peroxidation products level in hepatic tissues of guinea pigs. Activities of glutathione peroxidase (GPX) and glutathione reductase (GR) were enhanced in co-administered group. gamma-Glutamyl transpeptidase (GGT), a marker enzyme of alcohol induced toxicity, was also reduced, as was the glutathione content. This study suggests that the combined effect of Se+AA, provides protection against alcohol-induced oxidative stress as evidenced from the decreased levels of lipid peroxidation products and enhanced activities of scavenging enzymes.     

Degradation of Perennial Ryegrass Leaf and Stem Cell Walls by the Anaerobic Fungus Neocallimastix sp. Strain CS3b

The degradation of cell walls isolated from stems and leaves of perennial ryegrass by the anaerobic fungus Neocallimastix sp. strain CS3b was studied in a defined medium. The combined cellulose and hemicellulose fraction represented 53.1 (wt/wt) and 63.3% (wt/wt) of the dry weight of control grass leaf and stem cell walls, respectively. In both leaf and stem cell walls, glucose was the major neutral monosaccharide, followed by xylose, arabinose, and galactose. After 2 days of fermentation with Neocallimastix sp. strain CS3b, treated cell walls contained smaller amounts of neutral sugars compared with those of undigested cell walls. These results were more evident for glucose, xylose, and arabinose than for galactose. Furthermore, the sugar content of leaf cell walls decreased before a decline in the sugar content of stem cell walls was observed. Data from formate and hydrogen production indicated that the growth of Neocallimastix sp. strain CS3b was completed in 4 days in the culture system used. During this period, the fungus liberated about 95% of the fermentable sugars in untreated material. On a percentage basis, no significant differences were found in final extent of degradation of glucose, xylose, and arabinose. Galactose, however, was degraded to a lesser extent.     

A rapid chemical procedure for isolation and purification of chromosomal DNA from gram-negative bacilli

A rapid and simple method for preparing chromosomal DNA from gram-negative bacilli is presented. It is based on the alkaline (NaOH 0.03 m) lysis of cell walls. The resulting emulsion is purified by proteinase K (0.625 mg/g of wet wt), SDS, and the deproteinizing agent (chloroform isoamyl alcohol). The purity, molecular nature, and yield of DNA obtained by the present method are compared with those of DNA extracted by Marmur's procedure and a Marmur's modified procedure. We have developed and standardized this original method to isolate double-stranded DNA, free of proteins and RNA contamination and with a significantly higher yield of DNA than the two other methods. This procedure is particularly useful for strains with low growth and can be applied in every field concerned with DNA analysis.     

Copper and cobalt alter the cell wall composition of Cunninghamella blakesleeana

Cunninghamella blakesleeana was highly sensitive to Cu and Co on a medium containing NaNO3 as the sole nitrogen source. The nitrate reductive pathway was altered by Cu and Co, and NO-2 accumulated in the medium. Under conditions of Cu toxicity, the mycelium and the cell walls acquired a blue color, and most of the Cu was located in the cell walls, which differed in several aspects from cell walls derived from Co-containing or control cultures. At half-maximal growth inhibition by Cu (2.5 micrograms/mL or 39.3 microM) or Co (3.5 micrograms/mL or 59.4 microM), the mycelia contained 1.5 micrograms Cu or 1.0 microgram Co/mg dry tissue, respectively, but the isolated cell walls contained 33.5 micrograms Cu or 1.8 micrograms Co/mg dry cell wall. The phosphorous content of mycelia from Co-containing cultures was the same as that from control cultures, whereas that of mycelia from Cu-containing cultures contained 36% less. However, the phosphorous content of the cell walls from mycelia cultured in the presence of Cu or Co was two- and three-fold higher, respectively, than that of cell walls from control cultures. The cell walls of Cu-containing cultures contained significantly less hexosamine than the control cell walls, and chitin and chitosan were present in equal quantities. The cell walls of Co-containing cultures had the same amount of hexosamine as the control cell walls, but 88% of the hexosamine was present as chitosan and bound very little Co. The control cell walls contained approximately 60% chitosan.(ABSTRACT TRUNCATED AT 250 WORDS)     

The role of oxygen limitation in the formation of poly-β-hydroxybutyrate during batch and continuous culture of Azotobacter beijerinckii

Azotobacter beijerinckii was grown in ammonia-free glucose-mineral salts media in batch culture and in chemostat cultures limited by the supply of glucose, oxygen or molecular nitrogen. In batch culture poly-beta-hydroxybutyrate was formed towards the end of exponential growth and accumulated to about 74% of the cell dry weight. In chemostat cultures little poly-beta-hydroxybutyrate accumulated in organisms that were nitrogen-limited, but when oxygen limited a much increased yield of cells per mol of glucose was observed, and the organisms contained up to 50% of their dry weight of poly-beta-hydroxybutyrate. In carbon-limited cultures (D, the dilution rate,=0.035-0.240h(-1)), the growth yield ranged from 13.1 to 19.8g/mol of glucose and the poly-beta-hydroxybutyrate content did not exceed 3.0% of the dry weight. In oxygen-limited cultures (D=0.049-0.252h(-1)) the growth yield ranged from 48.4 to 70.1g/mol of glucose and the poly-beta-hydroxybutyrate content was between 19.6 and 44.6% of dry weight. In nitrogen-limited cultures (D=0.053-0.255h(-1)) the growth yield ranged from 7.45 to 19.9g/mol of glucose and the poly-beta-hydroxybutyrate content was less than 1.5% of dry weight. The sudden imposition of oxygen limitation on a nitrogen-limited chemostat culture produced a rapid increase in poly-beta-hydroxybutyrate content and cell yield. Determinations on chemostat cultures revealed that during oxygen-limited steady states (D=0.1h(-1)) the oxygen uptake decreased to 100mul h(-1) per mg dry wt. compared with 675 for a glucose-limited culture (D=0.1h(-1)). Nitrogen-limited cultures had CO(2) production values in situ ranging from 660 to 1055mul h(-1) per mg dry wt. at growth rates of 0.053-0.234h(-1) and carbon-limited cultures exhibited a variation of CO(2) production between 185 and 1328mul h(-1) per mg dry wt. at growth rates between 0.035 and 0.240h(-1). These findings are discussed in relation to poly-beta-hydroxybutyrate formation, growth efficiency and growth yield during growth on glucose. We suggest that poly-beta-hydroxybutyrate is produced in response to oxygen limitation and represents not only a store of carbon and energy but also an electron sink into which excess of reducing power can be channelled.     

Effects of sowing time and plant age on critical nitrogen concentrations in canola (Brassica napus L.)

Critical concentrations of NO₃-N in fresh petiole tissue and total N in the dried lamina were determined for the youngest mature leaf (YML) of field-grown canola. For dry matter yield of canola sown on 4 May, critical NO₃-N concentration in the YML petiole at the rosette stage (RS) was 1.46 mg/g fresh wt. At the flower-buds-visible stage (BV) it was 0.45 mg/g fresh wt. For seed yield the values were 1.72 and 0.53 mg/g fresh wt. Critical total N concentration in the YML lamina for dry matter yield were 69 mg/g dry wt. at RS and 57 at BV. For seed yield they were 71 and 59 mg/g dry wt. Critical NO₃-N concentrations in the YML petiole of canola sown on 30 May were reduced by 50%; critical total-N concentrations in the YML lamina were not reduced to the same extent. Despite the reductions in critical N concentrations in the YML, critical N fertilizer rates for vegetative growth and seed yield were unaffected by sowing date or plant growth stage.     

Changes in chemical composition of hydrophyte leaves during adaptation to aquatic environment

Leaf chemical composition of 19 hydrophytes was studied. The content of carbon, nitrogen, nonstructural carbohydrates, organic acids, minerals, and water was determined. Hydrophytes were shown to contain less carbon (below 410 mg/g dry wt in 60% species) than terrestrial plants. Hydrophytes and terrestrial plants did not differ in the nitrogen concentration in the leaves (33 and 29 mg/g dry wt, respectively). Hydrophytes were characterized by a low content of organic acids (40–90 mg/g dry wt in 60% species) and high content of mineral compounds (90–170 mg/g dry wt in 50% species). Total amount of nonstructural carbohydrates was similar in the leaves of hydrophytes and terrestrial plants (from 120 to 190 mg/g dry wt), but the proportions of various carbohydrate fractions differed substantially. In the hydrophyte leaves, the content of soluble carbohydrates was 2.4-fold lower, whereas the content of nonstructural polysaccharides 1.2-fold higher than in terrestrial plant leaves. Two groups of correlations between parameters of leaf chemical composition were distinguished: the contents of carbon, nitrogen, and soluble sugars were positively correlated, and the negative correlation was observed between these parameters and the amounts of mineral compounds, organic acids, water, and nonstructural polysaccharides. We concluded that hydrophyte leaf chemical composition reflects a specificity of plant adaptation to aquatic environment.     

Some methods for processing of single-cell protein

Methods for the production of protein concentrates, with a low content of nucleic acid, in kilogram quantities from yeast have been studied with the aid of equipment designed for operation on pilot-plant scale. The influence of drum drying and mechanical disintegration on the nutritive value of the yeast was also investigated. Drum drying and mechanical disintegration improved the nutritive value of the yeast but high extractability of protein and nucleic acid was only obtained after mechanical disintegration. Protein concentrates without and with cell walls were produced from mechanically disintegrated yeast. The different fractions which were obtained when separating cell walls and precipitating protein by heating at alkaline pH, were analyzed. After protein precipitation, about 90% of the RNA could be precipitated from the supernatant by addition of acid, giving a product containing 50% RNA of the dry weight. The protein precipitate obtained after cell wall separation had an RNA content of less than 2% and contained 70–l75% of the amino acids in the starting yeast material. Protein concentrates containing cell walls were produced by precipitating protein by heating at alkaline pH directly after mechanical disintegration. The content of RNA was about 2% and the yield of amino acids was 70–80%. It was found that the nutritive value of the protein concentrate was higher than that of the starting yeast material. To produce such a protein concentrate on a large scale, the process described can probably be employed.     

Changes in biochemical composition and energy utilization during metamorphosis of leptocephalous larvae of the bonefish (Albula)

Synopsis Energy use and changes in whole-body content of lipid, protein, nitrogen, carbohydrate and ash were followed during metamorphosis of leptocephalous larvae of the bonefish (Albula). During metamorphosis, which requires about 8–12 days, larvae lost about 3–4 mg of lipid, or about 50% of the total lipid content. Lipid levels, calculated on a dry weight basis, showed no discernible trends, with values ranging from 138–185 mg (g dry wt)^–1. Protein content was 8.4 mg per larva and showed no significant change. However, protein levels increased from 147 to 329 mg (g dry wt)^–1. Nitrogen content decreased slightly from about 3.5 to 3.2 mg per larva. A comparison of protein and nitrogen values, expressed as % dry weight, showed that, in larvae which were just beginning to metamorphose, 70% of the total nitrogen was non-protein nitrogen (NPN). The NPN decreased to 58% of the total nitrogen towards the end of metamorphosis. Carbohydrate content fell from about 3.5 to 0.6 mg per larva, which represents an 83% loss. Carbohydrate levels also fell from about 81 to 32 mg (g dry wt)^–1. In addition, most of the carbohydrate appears to be bound to protein. Ash content decreased by 52%, from 4.6 to 2.2 mg per larva. Caloric content fell slightly from about 182 to 141 calories per larva whereas caloric density showed no discernible trends, with values ranging from 4.180 to 4.725 kcal (g dry wt)^–1. These results indicate that metamorphosing leptocephali, which apparently do not feed, probably derive most of their energy requirements from metabolizing endogenous lipid and carbohydrate stores formed during the premetamorphic interval.     

Biochemical effects of garlic protein on lipid metabolism in alcohol fed rats

Garlic protein is a very good hypolipidemic agent. In the present study the water soluble protein fraction of garlic was investigated for its effect on hyperlipidemia induced by alcohol (3.76 g/kg. body wt./day). The hypolipidemic action is mainly due to an increase in cholesterol degradation to bile acids and neutral sterols and mobilization of triacyl glycerols in treated rats. Garlic protein (500 mg./kg body wt./day) showed significant hypolipidemic action comparable with a standard dose of gugu-lipid (50 mg./kg. body wt./day).     

The uptake of [3H]uridine into the nucleic acids of rat uterus during early pregnancy

1. Changes in content and uptake of [(3)H]uridine into the nucleic acids of rat uterus during the first 9 days of pregnancy were studied. 2. From day 6 implantation sites were separated from the rest of the uterine tissue for independent analysis. 3. Up to day 5 of pregnancy no changes were found in the total dry matter or in RNA and DNA content/unit dry matter nor in the RNA/DNA ratios. 4. From day 6, when implantation sites are visible, the water content of the implantation sites increased by 2-3%, and the RNA content/unit dry wt. and the RNA/DNA ratios increased. The DNA content/unit dry wt. did not increase in the implantation sites until day 8. 5. Uptake of [(3)H]uridine into the acid-soluble fraction of the tissues was markedly higher in implantation sites than in non-implantation sites. 6. Uptake of [(3)H]uridine into RNA was significantly increased on day 3 of pregnancy and again on day 5. 7. On days 6 and 7, the incorporation into RNA of implantation sites was significantly higher than in the remainder of the uterine tissue but decreased on days 8 and 9 to the same value as that of the normal tissue. 8. No change occurred in uptake into DNA until day 6, when there was an increase in uptake by the implantation sites. 9. It is suggested that the increase in RNA synthesis on day 3 is a preparation of the uterus for the onset of implantation on day 5, and that increased synthesis in implantation sites on days 6 and 7 is the elaboration of new RNA necessary for this early stage of pregnancy to commence.     

Measurement of concentrations of metabolites in adipose tissue and effects of insulin, alloxan-diabetes and adrenaline

1. Methods are described for the extraction and assay of ATP, ADP, AMP, glucose 6-phosphate, l-glycerol 3-phosphate and citrate in rat epididymal adipose tissue incubated in vitro for 1hr. At this time of incubation rates of glucose uptake and outputs of glycerol, free fatty acids, lactate and pyruvate were shown to be constant. 2. In fat pads incubated in medium containing glucose (3mg./ml.) and albumin (20mg./ml.) the concentrations (in mmumoles/g. wet wt.) were: ATP, 70; ADP, 36; AMP, 9.0; glucose 6-phosphate, 3.0; l-glycerol 3-phosphate, 3.3; citrate, 8.1. 3. The volume of intracellular water calculated from ([(3)H]water space-[(14)C]sorbitol space), ([(14)C]urea space-inulin space) and (weight loss on drying-[(14)C]sorbitol space) was 1.4ml./100g. wet wt. of tissue. The intracellular volume was not changed by insulin, alloxan-diabetes or adrenaline. 4. When compared in terms of mumoles/ml. of intracellular water the concentration of ATP in adipose tissue was less than in heart and diaphragm muscles. The concentrations of ADP and AMP were greater both in absolute terms and relative to ATP. Insulin, alloxan-diabetes and adrenaline had no significant effects on the concentrations of the adenine nucleotides in adipose tissue. 5. The concentration of glucose 6-phosphate was increased by insulin and lowered by alloxan-diabetes and adrenaline. The concentration of l-glycerol 3-phosphate was increased by insulin, unchanged by alloxan-diabetes and lowered by adrenaline. The concentration of citrate was increased by adrenaline and alloxan-diabetes and unchanged by insulin. 6. The effect of glucose concentration in the medium on rates of glucose uptake in adipose tissue from normal rats and alloxan-diabetic rats was investigated. The K(u) of glucose uptake was 29-44mg./100ml. and the V(max.) was 0.77mg./g. wet wt. of tissue/hr. Insulin increased the V(max.) and alloxan-diabetes diminished it, but neither agent significantly altered the K(u). 7. The significance of these results in relation to control of metabolism of adipose tissue is discussed.     

Growth and rutin production in hairy root cultures of buckwheat (Fagopyrum esculentum M.)

Buckwheat (Fagopyrum esculentum Moench.) is a potentially important source of rutin, a natural flavonoid with antihyperglycemic, antihypertensive, and antioxidative properties. To examine in vitro production of rutin, we established a hairy root culture of buckwheat by infecting leaf explants with Agrobacterium rhizogenes R1000, and tested the growth conditions and rutin production rates of these cultures. Ten hairy root clones were established; their growth and rutin production rates ranged from 233 to 312 (mg dry wt per 30 mL flask, and 0.8 to 1.2 (mg/g dry wt), respectively. Clone H8, which had high growth and rutin production rates (312 mg dry wt per 30 mL flask and 1.2 mg/g dry wt, respectively), was selected for further experiments. H8 showed maximal growth and rutin content at 30 days in culture in MS medium. Of four tested culture media, half-strength MS medium was found to induce the highest levels of growth (378 mg dry wt per 30 mL flask) and rutin production (1.4 mg/g dry wt) by clone H8. In contrast, supplementation with auxins (0.1-1 mg/l IAA, IBA and NAA) increased the growth rate, but had no significant effect on rutin production by H8. Collectively, these findings indicate that hairy root cultures of buckwheat culture could be a valuable alternative approach for rutin production.     

Studies on Tissue Culture ef Medicinal Plants VI. The Cost Question in the Cell Culture of Anisodus acutangulus

Both increased dry wt. of the culture cells and contents of hyoscyamine and scopolamine in the culture cells were decreased slightly (decreased about 1.2% and 2.9% respectively) when non-ionic water was used instead of redistilled water in the cell suspension culture of Anisodus acutangulus. But the non-ionic water could reduce costs largely, e.g. power cost etc. (can be reduced about 1030 yuantonne medium). Since non-ionic water can now be mass-produced easily, it suits the needs of industrial production in the future. The increased dry wt. of culture cells was a litter high and contents of two alkaloids was decreased slightly (about 5.4%) in the cell suspension culture of A. acutangulus when store confectioner’s sugar was used instead of sucrose. But the applied quantities of store confectioner’s sugar and costs could be reduce 50% and 88% respectively. The suspension cells of A. acutangulus contain rich amylase (average amylase activity was 4.37 mg reducing sugar/g, fresh wt./hour), so it could use starck as their only carbon source for the growth.     

Aluminium, iron and copper in human brain tissues donated to the Medical Research Council's Cognitive Function and Ageing Study

Aluminium, iron and copper are all implicated in the aetiology of neurodegenerative diseases including Alzheimer's disease. However, there are very few large cohort studies of the content of these metals in aged human brains. We have used microwave digestion and TH GFAAS to measure aluminium, iron and copper in the temporal, frontal, occipital and parietal lobes of 60 brains donated to the Cognitive Function and Ageing Study. Every precaution was taken to reduce contamination of samples and acid digests to a minimum. Actual contamination was estimated by preparing a large number of (170+) method blanks which were interspersed within the full set of 700+ tissue digests. Subtraction of method blank values (MBV) from tissue digest values resulted in metal contents in all tissues in the range, MBV to 33 μg g(-1) dry wt. for aluminium, 112 to 8305 μg g(-1) dry wt. for iron and MBV to 384 μg g(-1) dry wt. for copper. While the median aluminium content for all tissues was 1.02 μg g(-1) dry wt. it was informative that 41 brains out of 60 included at least one tissue with an aluminium content which could be considered as potentially pathological (> 3.50 μg g(-1) dry wt.). The median content for iron was 286.16 μg g(-1) dry wt. and overall tissue iron contents were generally high which possibly reflected increased brain iron in ageing and in neurodegenerative disease. The median content for copper was 17.41 μg g(-1) dry wt. and overall tissue copper contents were lower than expected for aged brains but they were commensurate with aged brains showing signs of neurodegenerative disease. In this study we have shown, in particular, the value of carrying out significant numbers of method blanks to identify unknown sources of contamination. When these values are subtracted from tissue digest values the absolute metal contents could be considered as conservative and yet they may still reflect aspects of ageing and neurodegenerative disease in individual brains.