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Biomedical subjects

A P Wilkinson

Publications and source records attributed to A P Wilkinson.

15 recordsLinked to original sources
PubMed2000

M-F interatomic distances and effective volumes of second and third transition series MF6- and MF6(2-) anions.

Synchrotron X-ray powder diffraction data (SPDD) for representative LiMF6 and Li2MF6 salts of the second and third transition series have provided unit-cell parameters and, from Rietveld analysis, M-F interatomic distances. M-F distances have also been obtained from X-ray single-crystal structural analyses of LiOsF6, Li2PtF6, and KRhF6. The LiMF6 all have the LiSbF6 structure type (space group R3). For M = Ta to Au the primitive unit cell volume decreases with increasing nuclear charge (Z), the volumes (sigma = 0.01 A3) being as follows: Ta, 111.26; Os, 102.42; Ir, 100.77; Pt, 99.62; and Au, 99.12 A3. A similar contraction, with increase in Z, occurs from Nb to Rh, the primitive cell volume (sigma = 0.01 A3) being: Nb, 110.92; Ru, 100.51; and Rh, 98.64 A3. For the TaF6- to AuF6- the M-F distances are not significantly different across the series, at approximately 1.87(1) A; also, Nb-F, Ru-F, and Rh-F = 1.86(1) A. In each series, the a and c values of the hexagonal-cell representation for the LiMF6 structure (separate layers of MF6- and Li+ stacked along c) change smoothly. As Z increases, a decreases and c increases. The variation in a, like the volume change, indicates that the size of MF6- is decreasing with Z. The variation in c suggests that the charge on the F-ligand is decreasing with Z. In the trirutile Li2MF6 series, M = Mo to Pd, the formula-unit volume decreases with Z(Mo, 100.92(6); Ru, 98.21(1); Rh, 97.43(1); Pd, 96.83(1) A3) and a shortening in M-F occurs (Mo-F = 1.936(4); Ru-F = 1.921(7); Rh-F = 1.910(7); Pd-F = 1.899(4) A). The less abundant data for MF6(2-) salts of the third transition series indicate similar trends. For both series, M-F distances of MF6(2-) are longer by 0.03-0.09 A than in MF6-.

Journal Article↗
PubMed2000

Effects of cationic charge on three-dimensional structures of intercalative complexes: structure of a bis-intercalated DNA complex solved by MAD phasing.

We characterize intercalative complexes as either "high charge" and "low charge". In low charge complexes, stacking interactions appear to dominate stability and structure. The dominance of stacking is evident in structures of daunomycin, nogalamycin, ethidium, and triostin A/echinomycin. By contrast in a DNA complex with the tetracationic metalloporphyrin CuTMPyP4 [copper (II) meso-tetra(N-methyl-4-pyridyl)porphyrin], electrostatic interactions appear to draw the porphyrin into the duplex interior, extending the DNA along its axis, and unstacking the DNA. Similarly, DNA complexes of tetracationic ditercalinium and tetracationic flexi-di show significant unstacking. Here we report x-ray structures of complexes of the tetracationic bis-intercalator D232 bound to DNA fragments d(CGTACG) and d(BrCGTABrCG). D232 is analogous to ditercalinium but with three methylene groups inserted between the piperidinium groups. The extension of the D232 linker allows it to sandwich four base pairs rather than two. In comparison to CuTMPyP4, flexi-di and ditercalinium, stacking interactions of D232 are significantly improved. We conclude that it is not sufficient to characterize intercalators simply by net charge. One anticipates strong electrostatic forces when cationic charge is focused to a small volume or region near DNA and so must consider the extent to which cationic charge is focused or distributed. In sum, ditercalinium, with a relatively short linker, focuses cationic charge more narrowly than does D232. So even though the net charges are equivalent, electrostatic charges are expected to be of greater structural significance in the ditercalinium complex than in the D232 complex.

Cations↗
PubMed1999

Monoclonal anti-idiotype antibody mimicking the pesticide binding site of cutinase: potential for broad specificity organophosphate recognition.

An anti-idiotype monoclonal antibody (Mab) able to bind the organophosphate pesticides, chlorfenvinphos (CFV), ethyl paraoxon, tetrachlorfenvinphos and demeton-s-methyl, has been produced using as immunogen a Mab which binds to the active site of cutinase. The principle of using an anti-idiotype antibody as the mimic of a site on a protein able to bind a group of ligands has, therefore, been demonstrated, and may have implications for future research on broad specificity immunoanalysis of groups of compounds.

Animals↗
PubMed1992

Heterologous gene expression in Aspergillus niger: a glucoamylase-porcine pancreatic prophospholipase A2 fusion protein is secreted and processed to yield mature enzyme.

The cDNA gene encoding porcine pancreatic prophospholipase A2 (proPLA2) was cloned into an Aspergillus niger expression vector downstream of the glucoamylase (glaA) gene promoter region. When this construct was transformed into A. niger, no detectable PLA2 was produced. Evidence was obtained showing that the PLA2 gene was transcribed and that PLA2 is extremely susceptible to both intracellular and extracellular proteases of A. niger, thus indicating that translation products would be rapidly degraded. By fusing the proPLA2-encoding sequence to the entire glaA gene, secreted yields of PLA2 up to 10 micrograms/ml were obtained from a transformed protease-deficient strain of A. niger. PLA2 was secreted in young cultures as a fusion protein, but in older cultures, it was processed from the glucoamylase carrier protein. Secreted PLA2 was shown to be enzymatically active and to have the correct N-terminal amino acid (aa) sequence, although another form of processed PLA2 was also produced. This form included two aa of the proregion from PLA2. The potential for improving yields of secreted heterologous proteins from A. niger still further is discussed.

Amino Acid Sequence↗
PubMed1990

Transplacental transfer of aflatoxin in humans.

This study quantified aflatoxin (AFB1, AFG1 and AFQ1) by enzyme-linked immunosorbent assay in human cord sera obtained at birth and in serum obtained immediately after birth from the mother. The subjects of the study were residents of Songkhla, Thailand. Of the 35 samples of cord sera, 17 (48%) contained aflatoxin in concentrations from 0.064 to 13.6, mean 3.1 nmol/ml. By comparison only two (6%) of 35 maternal sera contained aflatoxin (mean 0.62 nmol/ml). These results demonstrate transplacental transfer and concentration of aflatoxin by the feto-placental unit which may be of biological importance. Aflatoxins are mutagenic, carcinogenic and teratogenic and cause immunosuppression in animals. The implications of these findings are potentially profound and deserve further study.

Aflatoxin B1↗
PubMed1990

High serum concentrations of aflatoxin in Nepal as measured by enzyme-linked immunosorbent serum assay.

1 Sera from 28 Nepalis, both patients and workers in a hospital in Banepa, Nepal were examined for AFB1 by enzyme-linked immunosorbent assay (ELISA). 2 This assay, previously validated using spiked sera, provides a sensitive rapid determination of serum aflatoxin (B1, G1 and Q1). 3 All 28 sera were positive with concentrations from 60 pg ml-1 to 10 ng ml-1. 4 These results suggest that consumption of aflatoxin in Nepal is high (greater than 50-800 ng kg-1 d-1). 5 No reports of the degree of contamination of food, human consumption or body fluid concentrations of aflatoxin in Nepal have been previously published. 6 Aflatoxin may contribute to the development of hepatocellular carcinoma, which is probably a common tumour in Nepal.

Aflatoxins↗
PubMed1988

Measurement of aflatoxin in Nigerian sera by enzyme-linked immunosorbent assay.

Sera from 78 healthy men donating blood in Enugu, Nigeria were examined for aflatoxin by enzyme-linked immunosorbent assay (ELISA). Levels varied from less than 20 pg/ml to 3.1 ng/ml. The ELISA method is simple, sensitive and specific, and therefore well suited to the low-resource tropical environment. Aflatoxin is ingested in considerable amounts in Nigeria and many contribute, with hepatitis B, to the development of hepatocellular carcinoma.

Adolescent↗
PubMed1988

Analysis of UK sera for aflatoxin by enzyme-linked immunosorbent assay.

1. Aflatoxins are toxic, carcinogenic secondary fungal metabolites produced by certain moulds that commonly infest foods. Measurement of aflatoxins in human serum would give a direct measurement of exposure. 2. Twenty-seven serum samples from UK blood donors were found to contain aflatoxin levels not greater than 64 pmol/l (20 pg/ml) by an enzyme-linked immunosorbent assay. 3. These findings may indicate that present UK guideline tolerances for aflatoxin in imported food are effective in limiting human exposure to toxic aflatoxins in the UK diet, though further work would be needed to confirm this. In particular, sub-populations suspected of being at higher risk may need special considerations.

Aflatoxin B1↗
PubMed1984

Determination of myrosinase (thioglucoside glucohydrolase) activity by a spectrophotometric coupled enzyme assay.

The hexokinase/glucose-6-phosphate dehydrogenase coupled enzyme system was used to assay for plant thioglucoside glucohydrolase (myrosinase, EC 3.2.3.1) by measuring the rate of glucose released during hydrolysis of glucosinolates. This coupled assay was compared with two other assays for myrosinase: a pH-stat assay that measures the rate of acid released during glucosinolate hydrolysis, and a spectrophotometric assay in which the decrease in the absorbance at 227.5 nm is used to measure the disappearance of the substrate, 2-propenylglucosinolate (DSA assay). The coupled and pH-stat assays were found to give comparable activities and were linear with enzyme concentration over the range 0 to 30 micrograms. The DSA assay gave lower myrosinase activity in comparison to the coupled and pH-stat assays. This is due to the lower concentrations of substrate and activator (ascorbate) which must be used in the assay. The DSA assay was found to give a nonlinear relationship with enzyme concentration over the range 2 to 30 micrograms. For these reasons this assay was found to be unsatisfactory. The coupled assay was found to be more sensitive and more widely applicable than the pH-stat assay as a routine continuous assay for myrosinase activity.

Ascorbic Acid↗
PubMedDate not supplied

An ELISA method for the rapid and simple determination of aflatoxin in human serum.

A rapid, simple method for the determination of aflatoxin in human serum using anti-aflatoxin antisera in an indirect, double antibody microtitration plate enzyme-linked immunosorbent assay (ELISA) is described. Direct assay of serum necessitated the availability of a large sample volume, due to the presence of 'non-specific' interference. Addition of methanol to sera removed the interference enabling the analysis to be performed on small volumes (0.5 ml) of serum. The sensitivity of the assay is 20 pg ml-1. Total assay time, including sample preparation, is 4.5 h. No aflatoxin was found in any of the UK samples analysed.

Aflatoxins↗