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Biomedical subjects

M Holmer

Publications and source records attributed to M Holmer.

7 recordsLinked to original sources

Effects of tributyltin (TBT) on the seagrass Ruppia maritima.

The effects of tributyltin (TBT) on the seagrass Ruppia maritima were studied in two growth experiments. Plants were sampled at stations in Odense Fjord and Lunkebugten, Denmark, and replanted in reference sediment without TBT, reference sediment spiked with TBT, and in impacted sediment sampled in the highly TBT contaminated (7-57 microg kg (-1) dw) Odense Fjord. Plant performance was studied at weekly intervals for 3-4 weeks, by measuring net photosynthetic activity, respiration, relative growth rate (RGR) and number of leaves. Net photosynthetic activity in plants from spiked and impacted sediment was reduced by up to 60% relative to reference plants. Respiration both increased and decreased in response to TBT exposure, while RGR was generally lower in plants from contaminated sediments (reduced by 8-25%). The effects of spiked and impacted sediment differed between the experiments, which could be partly explained by the bioavailability of TBT in the two treatments, but also by adaptation of the plants from Odense Fjord to TBT. Measurements of enhanced TBT concentrations in the sediments in Odense Fjord suggest an impact of TBT on R. maritima is possible under in situ conditions.

Adaptation, Physiological↗

Synthesis of [Ru(phen)(2)dppz](2+)-tethered oligo-DNA and studies on the metallointercalation mode into the DNA duplex.

To explore the binding properties of [Ru(phen)(2)dppz](2+) complex (phen = 1,10-phenanthroline, dppz = dipyrido[3,2-a:2',3'-c]phenazine) in a sequence-specific manner in DNA duplex, it was tethered through the dppz ligand to a central position as well as both at the 3'- and 5'-ends of oligodeoxyribonucleotide (ODN). The middle [Ru(phen)(2)dppz](2+)-ODN tethered was resolved and isolated as four pure diastereomers, while the 3'- or 5'-[Ru(phen)(2)dppz](2+)-ODNs were inseparable on RP-HPLC. Thermal stability of the (Ru(2+)-ODN).DNA duplexes is found to increase considerably (DeltaT(m) = 12.8-23.4 degrees C), depending upon the site of the covalent attachment of the tethered [Ru(phen)(2)dppz](2+) complex, or the chirality of the [Ru(phen)(2)dppz](2+)-linker tethered at the middle of the ODN, compared to the unlabeled counterpart. Gross differences in CD between the [Ru(phen)(2)dppz](2+)-tethered and the native DNA duplexes showed that the global duplex conformation of the former has considerably altered from the B-type, but is still recognized by DNase I. The thermal melting studies, CD measurements, as well as DNase I digestion data, are interpreted as a result of intercalation of the dppz moiety, which is realized by threading of the Ru(phen)(2) complex part through the DNA duplex core. DNase I footprinting with four diastereomerically pure middle ([Ru(phen)(2)dppz](2+)-ODN).DNA duplexes furthermore showed that the tethered [Ru(phen)(2)dppz](2+)-linker chirality dictates the stereochemical accessibility of various phosphodiester moieties (around the intercalation site) toward the cleavage reaction by the enzyme. The diastereomerically pure ruthenium-modified duplexes, with the well-defined pi-stack, will be useful to explore stereochemistry-dependent energy- and electron-transfer chemistry to understand oxidative damage to the DNA double helix as well as the long-range energy- and electron-transfer processes with DNA as a reactant.

Circular Dichroism↗

An imaging system for monitoring receptive field dynamics.

The paper describes a computerized method, termed receptive field imaging (RFI), for the rapid mapping of multiple receptive fields and their respective sensitivity distributions. RFI uses random stimulation of multiple sites, in combination with an averaging procedure, to extract the relative contribution from each of the stimulated sites. Automated multi-electrode stimulation and recording, with spike detection and counting, are performed on-line by the RFI programme. Direct user interpretation of receptive field changes is made possible by a user-friendly graphic interface. A series of imaging experiments was carried out to evaluate the functional capacity of the system. RFI was tested on the receptive fields in the nociceptive withdrawal reflex (NWR) system in the rat. RFI replicates the results obtained with conventional methods and allows the display of receptive field dynamics induced by topical spinal cord application of morphine and naloxone on a minute-to-minute time scale. Data variance was estimated, and proved to be small enough to yield a stable representation of the receptive field, thereby achieving a high sensitivity in dynamic imaging experiments. The large number of stimulation and registration sites that can be monitored in parallel permits detailed network analysis of synaptic sets, corresponding to 'connection weights' between individual neurones.

Analysis of Variance↗

SCD5, a suppressor of clathrin deficiency, encodes a novel protein with a late secretory function in yeast.

Clathrin and its associated proteins constitute a major class of coat proteins involved in vesicle budding during membrane transport. An interesting characteristic of the yeast clathrin heavy chain gene (CHC1) is that in some strains a CHC1 deletion is lethal, while in others it is not. Recently, our laboratory developed a screen that identified five multicopy suppressors that can rescue lethal strains of clathrin heavy chain-deficient yeast (Chc - scd1-i) to viability. One of these suppressors, SCD5, encodes a novel protein of 872 amino acids containing two regions of repeated motifs of unknown function. Deletion of SCD5 has shown that it is essential for cell growth at 30 degrees C. scd5-delta strains carrying low copy plasmids encoding C-terminal truncations of Scd5p are temperature sensitive for growth at 37 degrees C. At the nonpermissive temperature, cells expressing a 338-amino acid deletion (Scd5P-delta 338) accumulate an internal pool of fully glycosylated invertase and mature alpha-factor, while processing and sorting of the vacuolar hydrolase carboxypeptidase Y is normal. The truncation mutant also accumulates 80- to 100-nm vesicles similar to many late sec mutants. Moreover, at 34 degrees C, overexpression of Scd5p suppresses the temperature sensitivity of a sec2 mutant, which is blocked at a post-Golgi step of the secretory pathway. Biochemical analyses indicate that approximately 50% of Scd5p sediments with a 100,000 x g membrane fraction and is associated as a peripheral membrane protein. Overall, these results indicate that Scd5p is involved in vesicular transport at a late stage of the secretory pathway. Furthermore, this suggests that the lethality of clathrin-deficient yeast can be rescued by modulation of vesicular transport at this late secretory step.

Amino Acid Sequence↗

Tax policy and the demand for health insurance.

Estimates of the price and income elasticities of demand for health insurance are derived from econometric estimates of the parameters of a discrete choice model of health insurance demand using data from a survey of plan choices among a sample of federal government employees. The estimated income elasticity (+0.01) is similar to most previous estimates. The estimated price elasticity (-0.16) is in absolute value much less than the earlier previous estimates and somewhat less than the more recent previous estimates. Possible causes of these differences in price elasticity estimates are discussed.

Community Participation↗

Bilirubin-protein interactions monitored by difference spectroscopy.

Difference spectroscopy is used to monitor bilirubin-protein interactions, to assess the residual binding capacity of proteins for bilirubin. A change in the difference spectra monitored at 482 nm is directly proportional to bound bilirubin up to a molar ratio of bilirubin to albumin of approximately 1; increasing bilirubin beyond the 1:1 molar ratio does not further change the difference spectra. After excess free bilirubin is added, the change in the difference spectrum is proportional to the residual binding capacity of the serum for bilirubin. The risk of kernicterus among neonates may be assessed by monitoring the residual bilirubin binding capacity of serum. This report summarizes our research effort leading to an assay method which requires only 40 microliter of serum and can be completed in less than 10 min.

Adult↗

Glutamine peptide does not improve gut barrier function and mucosal immunity in total parenteral nutrition.

In two separate experiments, three groups of 10 (first experiment) and 12 (second experiment) rats each were catheterized for the long term via the right jugular vein and fed one of the following dietary regimens for 7 days while in individual metabolism cages. The rat food control groups were fed powdered rat food. The total parenteral nutrition (TPN) formula for one group was enriched with the stable dipeptide L-alanyl-L-glutamine (ALA-GLN) equivalent to a 1% or 2% glutamine concentration. For the control TPN groups, the dipeptide was replaced by isonitrogenous amounts of alanine (ALA). In both experiments, bacterial translocation was promoted to a similar extent by the ALA-GLN and ALA TPN formulas compared with the rat food groups. Bacterial overgrowth in the cecum and intestinal atrophy, quantitated by the mucosal protein content, did not differ between ALA-GLN and ALA groups. The nitrogen balances of the three groups showed similar values in the second experiment. The secretory immunoglobulin A was measured in washings of the jejunum and ileum (soluble fraction) and in the homogenates of the gut mucosa (insoluble fraction). The secretory immunoglobulin A values of both fractions showed no difference between the ALA and ALA-GLN groups, irrespective of whether they were normalized for gut segment length or corrected for mucosal protein content. In conclusion from these results, the supplementation of TPN with glutamine does not seem to improve gut barrier function or mucosal immunity in unstressed rats.

Alanine↗