A mechanism of benzoic acid biosynthesis in plants and bacteria that mirrors fatty acid beta-oxidation.
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Biomedical subjects
Publications and source records attributed to A P Jarvis.
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Rapid oxidation and accretion of iron onto high surface area media has been investigated as a potential passive treatment option for ferruginous. net-alkaline minewaters. Two pilot-scale reactors were installed at a site in County Durham, UK. Each 2.0 m high cylinder contained different high surface area plastic trickling filter media. Ferruginous minewater was fed downwards over the media at various flow-rates with the objective of establishing the efficiency of iron removal at different loading rates. Residence time of water within the reactors was between 70 and 360 s depending on the flow-rate (1 and 12 l/min, respectively). Average influent total iron concentration for the duration of these experiments was 1.43 mg/l (range 1.08-1.84 mg/l; n = 16), whilst effluent iron concentrations averaged 0.41 mg/l (range 0.20-1.04 mg/l; n = 15) for Reactor A and 0.38 mg/l (range 0.11-0.93 mg/l; n = 16) for Reactor B. There is a strong correlation between influent iron load and iron removal rate. Even at the highest loading rates (approximately 31.6 g/day) 43% and 49% of the total iron load was removed in Reactors A and B, respectively. At low manganese loading rates (approximately 0.50-0.90 g/day) over 50% of the manganese was removed in Reactor B. Iron removal rate (g/m3/d) increases linearly with loading rate (g/day) up to 14 g/d and the slope of the line indicates that a mean of 85% of the iron is removed. In conclusion. it appears that the oxidation and accretion of ochre on high surface area media may be a promising alternative passive technology to constructed wetlands at certain sites.
Based on reversed-phase high-performance liquid chromatography (RP-HPLC) and atmospheric pressure chemical ionization (APCI) mass spectrometry, a HPLC-MS method was developed to permit the rapid qualitative and quantitative analysis of azadirachtin and related tetranortriterpenoids from seeds and tissue cultures of Neem (Azadirachta indica). APCI+ standard scanning mass spectra of the major Neem triterpenoids were recorded and utilized to select suitable ions for selected ion monitoring (SIM). Transitions for selective reaction monitoring (SRM) were based on MS-MS experiments. Using SIM, major Neem triterpenoids were detected in callus culture material and seed kernels of A. indica. The limit of detection for azadirachtin in extract samples (approximately 1 ng ml(-1) or 10 pg in SIM mode) was determined to be (with respect to injected absolute amounts) approximately 1000-times lower than values quoted in the literature for existing HPLC methods (approximately 200 ng ml(-1) or 10 ng). In addition to high sensitivity, the HPLC-MS method is able to tolerate minimal sample preparation and purification, dramatically reducing total analysis time.
Stable-isotope-labelled (2H6, 18O) 3-hydroxy-3-phenylpropanoic acid, a putative intermediate in the biosynthesis of benzoic acid (BA) and salicylic acid (SA) from cinnamic acid, has been synthesized and administered to cucumber (Cucumis sativus L.) and Nicotiana attenuata (Torrey). Analysis of the products by gas chromatography-mass spectrometry revealed incorporation of labelling into BA and SA, but not into benzaldehyde. In a separate experiment, 3-hydroxy-3-phenylpropanoic acid was found to be a metabolite of phenylalanine, itself the primary metabolic precursor of BA and SA. These data suggest that cinnamic acid chain shortening is probably achieved by beta-oxidation, and that the proposed "non-oxidative" pathway of side-chain degradation does not function in the biosynthesis of BA and SA, in cucumber and N. attenuata.
Fifty women in labour were allocated randomly to receive either air or saline to assist in the identification of the extradural space by the loss of resistance technique. A study volume of 4 ml of air or saline was used before 0.5% bupivacaine 8 ml and the spread of analgesia was followed for 30 min. The first segment blocked, time of onset, number of blocked segments and height of block were comparable in the two groups. At 30 min, there were eight patients with an unblocked segment in the air group, compared with two in the saline group (P less than 0.01). All unblocked segments were blocked subsequently by further doses of bupivacaine. We conclude that air is more likely than saline to produce unblocked segments in the initiation of extradural analgesia in labour.
We determined in 51 healthy patients undergoing body surface surgery the dose requirements for propofol, as part of a total i.v. anaesthesia technique with an alfentanil infusion. After premedication with temazepam, patients received alfentanil 50 micrograms kg-1 followed by an infusion of 50 micrograms kg-1 h-1. Patients were anaesthetized with a loading dose of propofol followed by a three-stage infusion designed to reach one of five preselected blood concentrations of propofol. The motor response to the initial surgical incision was noted and probit analysis was used to derive the ED50 (2.94 mg kg-1 h-1; 95% confidence limits: 2.35-3.37 mg kg-1 h-1) and ED95 (4.98 mg kg-1 h-1; 95% limits: 4.13-8.8 mg kg-1 h-1) for the final propotol infusion rate under these conditions. Whole blood concentration!of propofol at the time of the incision was related linearly to the inf!sion rate and the EC50 and EC95 (probit analysis) were derived as !.44 (95% confidence limits 0.62-1.87) and 4.05 (95% confidence lim!ts 2.78-30.5) micrograms ml-1, respectively. Post-operative recovery was!rapid, uncomplicated and uneventful. In a subgroup of eight patients,!the addition of clonidine 0.6 mg to the premedication significantly decreased the requirement for propofol (P less than 0.05) during surgery, but resulted in prolonged recovery times.
Total intravenous anesthesia by infusions of propofol and alfentanil may be associated with decreases in heart rate and blood pressure. The effects of two vagolytic agents on these hemodynamic changes were studied in 24 ASA physical status 1 patients undergoing body surface surgery. Patients were randomly allocated to receive atropine 10 micrograms/kg, glycopyrrolate, 5 micrograms/kg, or 0.9% sodium chloride, intravenously, 5 min before induction of anesthesia with loading doses of alfentanil, 50 micrograms/kg and propofol 1 mg/kg. Anesthesia was maintained with infusions of alfentanil 50 micrograms.kg-1.hr-1, and propofol 10 mg.kg-1.hr-1 for the first 10 min, 8 mg.kg-1.hr-1 for the next 10 min, and 6 mg.kg-1.hr-1 thereafter. Patients given glycopyrrolate before anesthesia had significantly higher arterial pressures than did patients receiving either atropine or saline, even though heart rates increased equally after glycopyrrolate and atropine.
The cell growth and monoclonal antibody production characteristics of two rat x mouse heterohybridoma cell lines, designated 187.1 and M1/9.3, were investigated using a biocompatible microencapsulation technology. Both cell lines, derived from the fusion of immunized rat spleen cells with either the NS1 or X63Ag8.653 myeloma cell lines, were found to reach a maximum intracapsular cell density of 1.3 to 1.5 X 10(7) cells/ml during a 27-d culture period. During this period, rat monoclonal antibody accumulated in the intracapsular space of both cultures to a final concentration of 2.0 to 2.8 mg/ml. Comparison of the concentration of rat monoclonal antibody in the extracapsular vs. the intracapsular space of both cultures indicated that significantly less than 1% of the antibody produced by the encapsulated hybridoma cells was capable of transiting the microcapsule membrane during the culture period. Due to the partition of the rat monoclonal antibody within the intracapsular space, the initial purity of the antibody harvested from 21-d microcapsule cultures of 187.1 and M1/9.3 cells was approximately 48 and 75% by weight, respectively. Analysis of the intracapsular protein by sodium dodecyl sulfoxide gel electrophoresis at different times during the culture period demonstrated that the principal contaminant associated with the unpurified antibody was bovine serum albumin.
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A new microencapsulation technology, developed for the encapsulation of living cells, has been demonstrated to be useful for the study of growth and differential gene expression using Friend erythroleukemic cells cultured at high cell densities. Using this technology, cultures of FL Clone 745 cells were encapsulated within semipermeable membranes composed of cross-linked alginic acid and poly-l-lysine. Cell growth studies measuring total cell number demonstrated an average generation time of 8.5 h in 5% (vol/vol) microcapsule cultures vs. 8.0 h in suspension cultures. Similar microcapsule cultures were serially propagated for more than 90 cell generations (13 sequential passages) with no significant change in this growth rate. In addition, final culture densities of greater than 1.0 X 10(8) cells/ml of intracapsular volume were attained using a 3% (vol/vol) microcapsule culture in conjunction with a standard refeeding schedule. Comparison of the level of dimethyl sulfoxide-induced hemoglobin production in suspension and microcapsule cultures demonstrated that the total amount of hemoglobin produced on a per cell basis was comparable in both systems. Due to the retention characteristics of the semipermeable membrane, the concentration of detergent-released hemoglobin, relative to other released protein, was approximately twofold higher in microcapsule cultures than in control suspension cultures.
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Using intraspecific hybrids, we have demonstrated the dominant nature of two phenotypic markers present in a mutant mouse 3T6 cell line, designated 3T6-VrB2. These are, resistance to virus infection (Vr) and semiconstitutive synthesis of interferon (IFsc). Hybrids were formed by polyethylene glycol-mediated fusion between 3T6-VrB2, or its parent 3T6, and 2TG0-13, a triply marked derivative of mouse 3T3 cells. When tested for the Vr marker, 3T6-VrB2 X 2TG0-13 hybrid clones displayed a level of resistance to virus infection which was equal to or greater than that of 3T6-VrB2. Similarly, when tested for the IFsc marker, these hybrid clones were found to possess the capacity to confer an interferon-induced antiviral state in mouse L929 cells upon cocultivation. By comparison, clones derived from 3T6 X 2TG0-13 fusions produced high levels of virus and failed to confer an interferon-induced antiviral state in L929 cells.
We describe the isolation and characterization of a virus-resistant mutant of murine 3T6 cells. The mutant, designated 3T6-VrB2, displays a high degree of resistance to infection by members of the toga-, rhabdo- and picornavirus classes. The level of this resistance to infection is similar to the parent 3T6 pretreated with approximately 100 lU/ml of interferon. Upon co-cultivation of 3T6-VrB2 cells with interferon-sensitive mouse cells, an antiviral state is induced in the latter cells as measured by a reduction of virus yield following infection. The nature of the induction is defined by a series of experiments using anti-mouse interferon antiserum. In the presence of this antiserum, the ability of the mutant to induce an antiviral state in interferon-sensitive mouse cells upon co-cultivation is eliminated. Additionally, growth of the mutant cells in the presence of this antiserum causes a reversal of the virus-resistant phenotype. Our results indicate that 3T6-VrB2 contains a mutation affecting the regulation of the murine interferon system such that the cell is engaged in the semiconstitutive synthesis of interferon.
Cytoplasmic extracts of untreated cultures of a virus-resistant mutant of mouse 3T6 cells, designated 3T6-VrB2, contain two double-stranded, RNA-activated enzyme activities associated with interferon action. These are the synthesis of a low molecular weight oligonucleotide inhibitor of cell-free protein synthesis from ATP, and the phosphorylation of a 67,000 dalton polypeptide by transfer of the gamma phosphate of ATP. Basal levels of both enzyme activities are detectable in extracts of untreated parental 3T6 cells, and are greatly enhanced upon interferon pretreatment. A procedure was developed, using a nonionic detergent to effect cell lysis, which allowed the analysis of the protein kinase activity from as few as 2 x 10(7) cells. Using this procedure, direct proportionalities were demonstrated between the concentration of interferon to which 3T6 cells were exposed, and both the level of protein kinase activity and the magnitude of the antiviral state were established in these cells. Furthermore, untreated cultures of 3T6-VrB2 exhibited both an antiviral state and an intracellular protein kinase activity equal to that of cultures of the parental 3T6 cells pretreated with a single concentration of mouse interferon.