scholarly journals Protein chromatography on adsorbents with hydrophobic and ionic groups. Some properties of N-(3-carboxypropionyl)aminodecyl-sepharose and its interaction with wheat-germ aspartate transcarbamoylase

1975 ◽  
Vol 151 (2) ◽  
pp. 281-290 ◽  
Author(s):  
R J Yon ◽  
R J Simmonds
Keyword(s):  
Wheat Germ ◽  
Fold Increase ◽  
Sodium Deoxycholate ◽  
Exchange Chromatography ◽  
Aspartate Transcarbamoylase ◽  
Protein Chromatography ◽  
Ionic Groups ◽  
High Concentrations ◽  
Group 2 ◽  
Derivatives Of

1. The charge state of two derivatives of Sepharose prepared by the CNBr activation method were studied by acid-base titration and by ion-exchange chromatography. Dodecyl-Sepharose exhibited cationic groups (21mumol/ml of settled gel; pKa=9.6) that were tentatively assigned to the coupling isourea group. 2. CPAD-Sepharose [N-(3-carboxypropionyl)aminodecyl-Sepharose] has anionic (carboxyl) groups (pKa=4.5) and cationic groups (pKa=9.6) in roughly equal concentrations (e coupling group. CPAD-Sepharose is slightly negatively charged at pH 7.0 and substantially negatively charged at pH 8.5. 3. The pKa values of dodecyl-Sepharose and CPAD-Sepharose are unaffected by a 100-fold increase in the concentration of KCl. 4. CPAD-Sepharose has considerable affinity for wheat-germ aspartate transcarbamoylase at pH 8.5 when the adsorbent and enzyme are both negatively charged. The interaction involves the C10 chain but is relatively moderate compared with C10 chains associated only with positive charge. 5. Desorption of the enzyme adsorbed to CPAD-Sepharose can be achieved by raising the pH to increase the electrostatic repulsion, or by introducing the detergent sodium deoxycholate. Acetone and butan-1-ol also weaken the adsorption at pH 8.5. 6. High concentrations of sodium acetate or sodium phosphate induced the enzyme to bind more tightly to CPAD-Sepharose. 7. These results are discussed in terms of a ‘repulsion-controlled’ model or hydrophobic chromatography.

scholarly journals Wheat-germ aspartate transcarbamoylase. The effect of ligands on the inactivation of the enzyme by trypsin and denaturing agents

1973 ◽  
Vol 131 (4) ◽  
pp. 699-706 ◽  
Author(s):  
Robert J. Yon
Keyword(s):  
Sodium Dodecyl Sulphate ◽  
Wheat Germ ◽  
Sodium Dodecyl ◽  
Aspartate Transcarbamoylase ◽  
Carbamoyl Phosphate ◽  
Low Concentrations ◽  
Alkaline Conditions ◽  
High Concentrations ◽  
Very High ◽  
Resistant State

In the absence of added ligands aspartate transcarbamoylase (EC 2.1.3.2) from wheat germ is inactivated fairly rapidly by trypsin, by heat (60°C), by highly alkaline conditions (pH11.3) and by sodium dodecyl sulphate. Addition of UMP alone, at low concentrations, decreases the rate of inactivation by each of these agents significantly. Carbamoyl phosphate alone does not alter the rate of inactivation by trypsin and by the detergent, but it antagonizes the effect of UMP in protecting the enzyme against these agents. These results have been interpreted to mean that two conformational states are reversibly accessible to the enzyme, namely an easily inactivated state favoured in the presence of carbamoyl phosphate and a more resistant state favoured in the presence of UMP. In the absence of ligands the enzyme is in the easily inactivated conformation. At very high concentrations l-aspartate also protects the enzyme but to a smaller extent than UMP. Some implications of these results are discussed.

scholarly journals The purification, composition and specificity of wheat-germ agglutinin

1973 ◽  
Vol 131 (1) ◽  
pp. 155-162 ◽  
Author(s):  
A. K. Allen ◽  
A. Neuberger ◽  
N. Sharon
Keyword(s):  
Binding Site ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Egg White ◽  
Exchange Chromatography ◽  
Hydroxyl Group ◽  
Anomeric Configuration ◽  
Egg White Lysozyme ◽  
Hen’S Egg ◽  
Derivatives Of

1. The purification of wheat-germ agglutinin from commercial wheat germ is described. By ion-exchange chromatography three active proteins (isolectins) were separated, one of which was examined in detail. 2. The amino acid composition is unusual, as 20% of residues are half-cystine and 21% are glycine. Unlike most lectins and contrary to previous reports, this protein is not a glycoprotein. 3. The efficiency of various saccharides as inhibitors of the agglutination reaction was investigated and from this the specificity of the binding site was inferred. Of monosaccharides, only derivatives of glucose with a 2-acetamido group and a free 3-hydroxyl group are effective inhibitors, and glycosides of either anomeric configuration are bound. Oligosaccharides are much more powerful inhibitors of agglutination than are monosaccharides. 4. It is proposed that the binding site consists of three or four subsites with differing specificities, in a cleft in the molecule resembling that proposed for hen's-egg-white lysozyme.

Potential Contribution of Nutrients and Polycyclic Aromatic Hydrocarbons from the Creeks of Cootes Paradise Marsh

1996 ◽  
Vol 31 (3) ◽  
pp. 485-504 ◽  
Author(s):  
Patricia Chow-Fraser ◽  
Barb Crosbie ◽  
Douglas Bryant ◽  
Brian McCarry
Keyword(s):  
Aromatic Hydrocarbons ◽  
Laboratory Experiments ◽  
Dry Weight ◽  
Common Source ◽  
Potential Contribution ◽  
Nitrogen And Phosphorus ◽  
Engine Combustion ◽  
Polycyclic Aromatic ◽  
High Concentrations ◽  
Derivatives Of

Abstract During the summer of 1994, we compared the physical and nutrient characteristics of the three main tributaries of Cootes Paradise: Spencer, Chedoke and Borer’s creeks. On all sampling occasions, concentrations of CHL α and nutrients were always lowest in Borer’s Creek and highest in Chedoke Creek. There were generally 10-fold higher CHL α concentrations and 2 to 10 times higher levels of nitrogen and phosphorus in Chedoke Creek compared with Spencer Creek. Despite this, the light environment did not differ significantly between Spencer and Chedoke creeks because the low algal biomass in Spencer Creek was balanced by a relatively high loading of inorganic sediments from the watershed. Laboratory experiments indicated that sediments from Chedoke Creek released up to 10 µg/g of soluble phosphorus per gram (dry weight) of sediment, compared with only 2 µg/g from Spencer Creek. By contrast, sediment samples from Spencer Creek contained levels of polycyclic aromatic hydrocarbon that were as high as or higher than those from Chedoke Creek, and much higher than those found in Borer’s Creek. The distribution of normalized PAH concentrations suggests a common source of PAHs in all three tributaries, most likely automobile exhaust, since there were high concentrations of fluoranthene and pyrene, both of which are derivatives of engine combustion.

scholarly journals THE RATE OF BACTERICIDAL ACTION OF PENICILLIN IN VITRO AS A FUNCTION OF ITS CONCENTRATION, AND ITS PARADOXICALLY REDUCED ACTIVITY AT HIGH CONCENTRATIONS AGAINST CERTAIN ORGANISMS

1948 ◽  
Vol 88 (1) ◽  
pp. 99-131 ◽  
Author(s):  
Harry Eagle ◽  
A. D. Musselman
Keyword(s):  
Bacterial Species ◽  
Fold Increase ◽  
Bactericidal Effect ◽  
Penicillin G ◽  
Bacterial Suspension ◽  
Maximal Effect ◽  
Maximal Rate ◽  
Effective Level ◽  
High Concentrations

1. The concentrations of penicillin G which (a) reduced the net rate of multiplication, (b) exerted a net bactericidal effect, and (c) killed the organisms at a maximal rate, have been defined for a total of 41 strains of α- and ß-hemolytic streptococci, Staphylococcus aureus and Staphylococcus albus, Diplococcus pneumoniae, and the Reiter treponoma. 2. The concentration which killed the organisms at a maximal rate was 2 to 20 times the minimal effective level ("sensitivity" as ordinarily defined). With some organisms, even a 32,000-fold increase beyond this maximally effective level did not further increase the rate of its bactericidal effect. However, with approximately half the strains here studied (all 4 strains of group B ß-hemolytic streptococci, 4 of 5 group C strains, 5 of 7 strains of Streptococcus fecalis, 2 of 4 other α-hemolytic streptococci, and 4 of 9 strains of staphylococci), when the concentration of penicillin was increased beyond that optimal level, the rate at which the organisms died was paradoxically reduced rather than increased, so that the maximal effect was obtained only within a relatively narrow optimal zone. 3. There were marked differences between bacterial species, and occasionally between different strains of the same species, not only with respect to the effective concentrations of penicillin, but also with respect to the maximal rate at which they could be killed by the drug in any concentration. Although there was a rough correlation between these two factors, there were many exceptions; individual strains affected only by high concentrations of penicillin might nevertheless be killed rapidly, while strains sensitive to minute concentrations might be killed only slowly. 4. Within the same bacterial suspension, individual organisms varied only to a minor degree with respect to the effective concentrations of penicillin. They varied strikingly, however, in their resistance to penicillin as measured by the times required to kill varying proportions of the cells.

scholarly journals Thunderstorm outflows preceding epidemics of asthma during spring and summer

Thorax ◽  
2001 ◽  
Vol 56 (6) ◽  
pp. 468-471
Author(s):  
G B Marks ◽  
J R Colquhoun ◽  
S T Girgis ◽  
M Hjelmroos Koski ◽  
A B A Treloar ◽  
...  
Keyword(s):  
Pollen Grains ◽  
Ground Level ◽  
Fold Increase ◽  
Underlying Mechanism ◽  
Detailed Examination ◽  
Asthma Exacerbations ◽  
Eastern Australia ◽  
The Difference ◽  
High Concentrations ◽  
Thunderstorm Outflow

BACKGROUNDA study was undertaken to assess the importance of thunderstorms as a cause of epidemics of asthma exacerbations and to investigate the underlying mechanism.METHODSA case control study was performed in six towns in south eastern Australia. Epidemic case days (n = 48) and a random sample of control days (n = 191) were identified by reference to the difference between the observed and expected number of emergency department attendances for asthma. The occurrence of thunderstorms, their associated outflows and cold fronts were ascertained, blind to case status, for each of these days. In addition, the relation of hourly pollen counts to automatic weather station data was examined in detail for the period around one severe epidemic of asthma exacerbations. The main outcome measure was the number of epidemics of asthma exacerbations.RESULTSThunderstorm outflows were detected on 33% of epidemic days and only 3% of control days (odds ratio 15.0, 95% confidence interval 6.0 to 37.6). The association was strongest in late spring and summer. Detailed examination of one severe epidemic showed that its onset coincided with the arrival of the thunderstorm outflow and a 4–12 fold increase in the ambient concentration of grass pollen grains.CONCLUSIONSThese findings are consistent with the hypothesis that some epidemics of exacerbations of asthma are caused by high concentrations of allergenic particles produced by an outflow of colder air, associated with the downdraught from a thunderstorm, sweeping up pollen grains and particles and then concentrating them in a shallow band of air at ground level. This is a common cause of exacerbations of asthma during the pollen season.

scholarly journals Effects of lipids on nucleotide inhibition of wheat-germ aspartate transcarbamoylase: evidence of an additional level of control?

1996 ◽  
Vol 313 (2) ◽  
pp. 669-673 ◽  
Author(s):  
Ashan KHAN ◽  
Babur Z. CHOWDHRY ◽  
Robert J. YON
Keyword(s):  
Fatty Acids ◽  
Wheat Germ ◽  
Hill Coefficient ◽  
Aspartate Transcarbamoylase ◽  
Kinetic Effect ◽  
Carbamoyl Phosphate ◽  
Allosteric Mechanism ◽  
Additional Level ◽  
Nucleotide Inhibition ◽  
Metabolic Significance

Wheat-germ aspartate transcarbamoylase, a monofunctional trimer, is strongly inhibited by uridine 5ʹ-monophosphate (UMP), which shows kinetic interactions with the substrate, carbamoyl phosphate, suggesting a classical allosteric mechanism of regulation. Inhibition of the purified enzyme by UMP was amplified in the presence of a variety of ionic lipids at concentrations low enough to preclude denaturation. In the absence of UMP, most of these compounds had no kinetic effect or were slightly activating. Two phospholipids did not show the effect. In a homologous series of fatty acids (C6-C16), the potentiating effect was only seen with homologues greater than C8, reaching a maximum at C12. The effect of dodecanoate (C12) on kinetic cooperativity (UMP as variable ligand) was studied. At each of several fixed concentrations of carbamoyl phosphate, dodecanoate had a pronounced effect on the half-saturating concentration of UMP, which was reduced by about half in every case, indicating substantially tighter binding of UMP. However, dodecanoate had relatively little effect on the kinetic Hill coefficient for the cooperativity of UMP. The possible metabolic significance of these effects is discussed.

scholarly journals The effect of copper excess on iron metabolism in sheep

1978 ◽  
Vol 170 (1) ◽  
pp. 137-143 ◽  
Author(s):  
E C Theil ◽  
K T Calvert
Keyword(s):  
Iron Metabolism ◽  
Iron Concentration ◽  
Transferrin Saturation ◽  
Fold Increase ◽  
Copper Treatment ◽  
Excess Copper ◽  
High Concentrations ◽  
Copper Excess ◽  
Effect Of Copper ◽  
Haemoglobin Content

Sheep were treated with large amounts of copper (20 mg of CuSO4,5H2O/kg body wt. per day) for 9 weeks to examine the effect of copper excess on iron metabolism. In addition to confirming that massive haemolysis and accumulation of copper occurs in the liver, kidney and plasma after 7 weeks of exposure to excess copper, it was observed that excess copper produced an increased plasma iron concentration and transferrin saturation within 1 week. Further, iron preferentially accumulated in the spleen between 4 and 6 weeks of copper treatment, producing 3-fold increases in the iron content of both the ferritin and non-ferritin fractions. A 3-4 fold increase was also observed in the amount of ferritin that could be isolated from the spleen. The copper treatment had little or no effect on the concentration of iron in the liver and bone marrow. The following properties of erythrocytes were also unaffected by copper treatment: size, haemoglobin content and pyruvate kinase activity, although the erythrocyte concentration of copper increased after 6 weeks. Copper accumulated in the spleen between 6 and 9 weeks, probably owing to the phagocytosis of erythrocytes containing high concentrations of copper. The data suggest that copper excess influences iron metabolism, initially by causing a compensated haemolytic anaemia, and later by interfering with re-utilization of iron from ferritin in the reticuloendothelial cells of the spleen.

scholarly journals Derivation of nominal verbs with the suffix -(i)auti, that belong to the categorial value ʿto be something/ someone or such as what is denoted by the base word’

2012 ◽  
pp. 1-19
Author(s):  
Anželika Gaidienė
Keyword(s):  
Electronic Version ◽  
Base Word ◽  
Standard Language ◽  
Part Of Speech ◽  
First Category ◽  
Electronic Dictionary ◽  
Group 2 ◽  
Derivatives Of ◽  
Living Being

  The article deals with nominal verbs with the suffix -(i)auti that have common categorical meaning ‘to be something/someone or such as what is denoted by the base word.' The object of the research consists of 239 verbs found in the electronic Dictionary of the Contemporary Lithuanian Language and 236 corresponding verbs from the electronic version of the Dictionary of the Lithuanian Language. According to the derivational meaning (darybos reikšmė), the verbs under investigation are classified into two groups: 1) verbs that mean ‘to be what is denoted by the base word' (they constitute 83% of all verbs that are analysed here) and 2) verbs, meaning ‘to be such as denoted by the base word' (17%). The verbs of the first category are further subdivided into four subclasses: 1) ‘to be someone denoted by the base word (not in the sense of work)' (they make up 69% of all the verbs of the first group), 2) ‘to work as what is denoted by the base word' (25%), 3) ‘to behave as behaves a person or another living being denoted by the base word' (5%) and 4) verbs denoting ‘states, processes that arise from the thing denoted by the base word' (only 1%).The derivatives of these subclasses are further divided according to the part of speech of the base words (nouns, adjectives or numerals) and the type of derivation (simple forms or derivatives: suffix, prefix, inflection or compounds). The investigation carried out has demonstrated that the verbs derived from nominals by adding the suffix -(i)auti which belong to the category meanings ‘to be something/someone or such as what is denoted by the base word' in standard language have few suffix and stem variants, they are not very characteristic of root vowel and consonant change, their base words do not vary a lot. 

Abstract 97: Increased Plasma Level of ApoCIII-rich Electronegative HDL May Contribute to Cognitive Impairment in Alzheimer’s Disease

Stroke ◽  
2017 ◽  
Vol 48 (suppl_1) ◽  
Author(s):  
Hua-Chen Chan ◽  
Hsiao-Ting Lu ◽  
Mei-Chuan Chou ◽  
Ming-Hsien Tsai ◽  
Wei-Hsiang Chen ◽  
...  
Keyword(s):  
Cognitive Impairment ◽  
Cholesterol Efflux ◽  
Chemical Properties ◽  
Density Lipoprotein ◽  
Fold Increase ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Paraoxonase 1 ◽  
Cholesterol Efflux Capacity ◽  
Tnf Α

Background: High-density lipoprotein (HDL), the only lipoprotein class that can cross the blood brain barrier bidirectionally, is positively associated with cognitive functions. To delineate HDL’s role in Alzhenimer’s disease (AD), we analyzed the chemical properties of plasma HDL from AD and healthy normal adult (control) subjects. Methods and results: By using anion-exchange chromatography, we divided HDL into 5 increasingly electronegative subfractions, H1-H5. Compared to the control cohort (4.24±3.22%; n=20), HDL from AD patients (23.48±17.83%; n=30) had a 5.5-fold increase of H5 ( P <0.001; Figure ), accompanied by a decreased protein/lipid ratio attributed to a significant reduction of albumin essential for prevention of amyloid beta (Aβ) aggregation. As determined by LC/MS E and ProteinLynx Global SERVER (PLGS), AD-HDL was had a rich content of apolipoprotein (apo)CIII, but diminished amounts of sphingosine-1-phosphate (S1P)-associated apoM and antioxidative paraoxonase 1 (PON1). Exposure of murine RAW 264.7 macrophages to H5 induced vibrant expression of ganglioside GM1 in colocalization with apoCIII on lipid rafts, alongside a concomitant increase of TNF-α detectable in the cultured medium ( Figure ). LC/MS E examination localized posttranslational oxidation exclusively in ApoA1 residues of H5 in AD-HDL, which exhibited a compromised cholesterol efflux capacity. Conclusions: Plasma HDL from AD patients has a high proportion of H5, an apoCIII-rich electronegative HDL subfraction. The associated reduction in functional (albumin, S1P, apoM) and increase in proinflammatory (apoCIII, PON1, TNF-α) components may favor Aβ assembly and neuroinflammation. Additionally, a compromised cholesterol-efflux capacity of AD-HDL may also contribute to vascular cognitive impairment.

Abstract TP94: Mesenchymal Stromal Cells Behave Differently When Exposed to Medications Commonly Prescribed to Stroke Patients

Stroke ◽  
2017 ◽  
Vol 48 (suppl_1) ◽  
Author(s):  
Nikunj Satani ◽  
Bing Yang ◽  
Duyen M Nghiem ◽  
Xiaopei Xi ◽  
Adrian P Gee ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
High Glucose ◽  
Fold Increase ◽  
Stroke Recovery ◽  
Stroke Patients ◽  
Healthy Humans ◽  
Tnf Α ◽  
High Concentrations ◽  
And Function

Background: As a promising investigational therapy for stroke recovery, mesenchymal stromal cells (MSCs) are in various stages of clinical trials. MSCs may promote recovery through cytokine release and immunomodulation. Stroke patients typically are treated with antiplatelets and medications for hypertension and hyperlipidemia. We explored the effect of commonly prescribed drugs at physiological concentrations on MSCs. Methods: Clinical grade bone marrow MSCs from healthy donor at passage 2 were thawed and re-suspended in serum free media. Monocytes (Mo) were isolated from peripheral blood of healthy humans. MSCs and Mo were cultured alone as well as in co-culture and exposed to simvastatin, atenolol, losartan, captopril, or aspirin. They were also exposed to high glucose (upto 40mM) to simulate hyperglycemia. At 24 hours of incubation, media was collected and TNF-α concentration was measured, as an index of immunomodulation of Mo by MSCs. Cell viability was also measured (using MTT assay and flow cytometry). Results: There were significant effects of all drugs on viability of MSCs but with no impact on Mo. More importantly, Losartan (dose independent), Simvastatin and Atenolol (dose-dependent) reduced the viability of MSCs even at the pharmacologically relevant concentrations (Fig 1). High glucose had no effect on viability of MSCs or Mo. TNF-α secretion from co-culture of MSCs and Mo at 24 hours showed differences at very high doses of aspirin (2-fold increase), atenolol (0.5 fold decrease), and glucose (0.5 fold decrease) (data not shown). However, these high concentrations are unlikely to be achieved pharmacologically in plasma of patients treated with these drugs. Conclusion: Exposure of MSCs to clinically relevant drugs can alter their viability and function. Our results suggest that stroke trials involving use of intravenous MSCs should consider the differential impact of commonly prescribed medications on MSCs function.

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