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K J Willis

Publications and source records attributed to K J Willis.

At least 19 recordsLinked to original sources

What is natural? The need for a long-term perspective in biodiversity conservation.

Ecosystems change in response to factors such as climate variability, invasions, and wildfires. Most records used to assess such change are based on short-term ecological data or satellite imagery spanning only a few decades. In many instances it is impossible to disentangle natural variability from other, potentially significant trends in these records, partly because of their short time scale. We summarize recent studies that show how paleoecological records can be used to provide a longer temporal perspective to address specific conservation issues relating to biological invasions, wildfires, climate change, and determination of natural variability. The use of such records can reduce much of the uncertainty surrounding the question of what is "natural" and thereby start to provide important guidance for long-term management and conservation.

Animals↗

Sea lice treatments on salmon farms have no adverse effects on zooplankton communities: a case study.

The long-term effects of the sea lice treatment products Excis and Slice on zooplankton communities in a Scottish sea loch were investigated at a commercially operating salmon farm over 31 months. Cypermethrin and emamectin benzoate are the active ingredients in Excis and Slice respectively, which are widely used to control ectoparasitic sea lice on farmed salmon. Excis and Slice treatments did not cause basin-wide effects on the zooplankton community. For both formulations, no adverse affects on zooplankton were detected, instead observed changes in zooplankton abundance and community composition displayed natural seasonal cycles of abundance. Water column concentrations of cypermethrin and emamectin benzoate following sea lice treatments at the fish farm were predicted using models. Cypermethrin concentrations of 3000 ng/l were predicted for short periods immediately after each cage treatment assuming no particle adhesion. The 3-h and 24-h Environmental Quality Standards (EQS) were exceeded for 10 h and 32 h respectively on the second day when five cages were treated. However, cypermethrin concentrations higher than 0.5 ng/l (24-h EQS) were predicted to occur over <2% of the total basin area on each treatment day. The 3-h EQS (16 ng/l) was exceeded in <0.3% of the basin on each treatment day. The concentration of soluble emamectin benzoate present in the water column was predicted from modelled deposition footprints and sediment concentrations to be of order 10(-3) ng/l. Predicted concentrations of both chemicals were generally lower than those causing toxicity to copepods in previous laboratory studies and further support the results of this field study that environmental concentrations of Excis and Slice do not adversely impact zooplankton communities.

Animals↗

Holocene biomass burning and global dynamics of the carbon cycle.

Fire regimes have changed during the Holocene due to changes in climate, vegetation, and in human practices. Here, we hypothesise that changes in fire regime may have affected the global CO2 concentration in the atmosphere through the Holocene. Our data are based on quantitative reconstructions of biomass burning deduced from stratified charcoal records from Europe, and South-, Central- and North America, and Oceania to test the fire-carbon release hypothesis. In Europe the significant increase of fire activity is dated approximately 6000 cal. yr ago. In north-eastern North America burning activity was greatest before 7500 years ago, very low between 7500-3000 years, and has been increasing since 3000 years ago. In tropical America, the pattern is more complex and apparently latitudinally zonal. Maximum burning occurred in the southern Amazon basin and in Central America during the middle Holocene, and during the last 2000 years in the northern Amazon basin. In Oceania, biomass burning has decreased since a maximum 5000 years ago. Biomass burning has broadly increased in the Northern and Southern hemispheres throughout the second half of the Holocene associated with changes in climate and human practices. Global fire indices parallel the increase of atmospheric CO2 concentration recorded in Antarctic ice cores. Future issues on carbon dynamics relatively to biomass burning are discussed to improve the quantitative reconstructions.

Americas↗

Did dinosaurs invent flowers? Dinosaur-angiosperm coevolution revisited.

Angiosperms first appeared in northern Gondwana during the Early Cretaceous, approximately 135 million years ago. Several authors have hypothesised that the origin of angiosperms, and the tempo and pattern of their subsequent radiation, was mediated by changes in the browsing behaviour of large herbivorous dinosaurs (sauropods and ornithischians). Moreover, the taxonomic and ecological radiation of angiosperms has been associated with the evolution of complex jaw mechanisms among ornithischian dinosaurs. Here, we review critically the evidence for dinosaur-angiosperm interactions during the Cretaceous Period, providing explicit spatiotemporal comparisons between evolutionary and palaeoecological events in both the dinosaur and angiosperm fossil records and an assessment of the direct and indirect evidence for dinosaur diets. We conclude that there are no strong spatiotemporal correlations in support of the hypothesis that dinosaurs were causative agents in the origin of angiosperms; however, dinosaur-angiosperm interactions in the Late Cretaceous may have resulted in some coevolutionary interactions, although direct evidence of such interactions is scanty at present. It is likely that other animal groups (insects, arboreal mammals) had a greater impact on angiosperm diversity during the Cretaceous than herbivorous dinosaurs. Elevated levels of atmospheric CO2 might have played a critical role in the initial stages of the angiosperm radiation.

Animals↗

Preparation of enantiomerically pure L-7-azatryptophan by an enzymatic method and its application to the development of a fluorimetric activity assay for tryptophanyl-tRNA synthetase.

The reaction of D,L-7-azatryptophan (D,L-7AW) with tryptophanyl-tRNA synthetase (TrpRS), adenosine triphosphate (ATP), and Mg2+ in the presence of inorganic pyrophosphatase results in the formation of a highly fluorescent l-7AW-adenylate complex. Detection of this complex is based on its enhanced fluorescence at 315 nm excitation and 360 nm emission after the addition of ATP. This stereoselective reaction was used to develop an activity assay for TrpRS using commercially available racemic D,L-7AW. The assay can be used to determine the activity of TrpRS from samples which contain less than 1 nmol of enzyme in 250 microL of sample. Thus the enzyme activity can be assessed without resorting to a radioactive assay of tRNATrp acylation. A secondary use of the stereoselective assay was for confirming the presence of pure L-7AW, D-7AW, or mixtures of the two enantiomers. D-7AW and L-7AW were prepared by reacting D,L-7AW with chloroacetic anhydride to form N-chloroacetyl-D,L-7AW (ClAc-7AW) followed by stereospecific proteolytic digestion of ClAc-7AW using carboxypeptidase A to produce the free L-7AW. The L-7AW could be separated from unreacted N-chloroacetyl-7AW by reverse-phase HPLC. The TrpRS-based assay was able to unambiguously discriminate between the two enantiomers of 7AW. The assay was then used to identify which enantiomer of 7AW was present in resolved fractions of the tripeptide L-lysyl-D,L-7-azatryptophyl-L-lysine. Digestion of the resolved tripeptides with protease enzymes produced the free L or D enantiomer of 7AW, which was easily identified using the TrpRS assay procedure.

Adenosine Triphosphate↗

Naphthalene sulfonate polymers with CD4-blocking and anti-human immunodeficiency virus type 1 activities.

PIC 024-4 and PRO 2000 are naphthalene sulfonate polymers that bind to CD4 with nanomolar affinity and block binding of gp120. Both have activity against human immunodeficiency virus type 1 in H9 cells, peripheral blood mononuclear cells, and primary monocyte/macrophages, are synergistic with zidovudine, and do not inhibit tetanus toxoid-stimulated T-cell proliferation at anti-human immunodeficiency virus type 1 concentrations.

Animals↗

Conformation and function of the N-linked glycan in the adhesion domain of human CD2.

The adhesion domain of human CD2 bears a single N-linked carbohydrate. The solution structure of a fragment of CD2 containing the covalently bound high-mannose N-glycan [-(N-acetylglucosamine)2-(mannose)5-8] was solved by nuclear magnetic resonance. The stem and two of three branches of the carbohydrate structure are well defined and the mobility of proximal glycan residues is restricted. Mutagenesis of all residues in the vicinity of the glycan suggests that the glycan is not a component of the CD2-CD58 interface; rather, the carbohydrate stabilizes the protein fold by counterbalancing an unfavorable clustering of five positive charges centered about lysine-61 of CD2.

Acetylglucosamine↗

Probing alpha-helical secondary structure at a specific site in model peptides via restriction of tryptophan side-chain rotamer conformation.

The relationship between alpha-helical secondary structure and the fluorescence properties of an intrinsic tryptophan residue were investigated. A monomeric alpha-helix forming peptide and a dimeric coiled-coil forming peptide containing a central tryptophan residue were synthesized. The fluorescence parameters of the tryptophan residue were determined for these model systems at a range of fractional alpha-helical contents. The steady-state emission maximum was independent of the fractional alpha-helical content. A minimum of three exponential decay times was required to fully describe the time-resolved fluorescence data. Changes were observed in the decay times and more significantly, in their relative contributions that could be correlated with alpha-helix content. The results were also shown to be consistent with a model in which the decay times were independent of both alpha-helix content and emission wavelength. In this model the relative contributions of the decay time components were directly proportional to the alpha-helix content. Data were also analyzed according to a continuous distribution of exponential decay time model, employing global analysis techniques. The recovered distributions had "widths" that were both poorly defined and independent of peptide conformation. We propose that the three decay times are associated with the three ground-state chi 1 rotamers of the tryptophan residue and that the changes in the relative contributions of the decay times are the result of conformational constraints, imposed by the alpha-helical main-chain, on the chi 1 rotamer populations.

Amino Acid Sequence↗

Interaction with model membrane systems induces secondary structure in amino-terminal fragments of parathyroid hormone related protein.

The secondary structure of amino terminal fragments of human parathyroid hormone related protein (PTHrP) in aqueous solution, in trifluoroethanol solution and in the presence of model membrane systems has been studied by circular dichroism (CD) spectroscopy. Far-UV CD spectra of PTHrP 1-40, 1-34 amide, 7-34 amide and 1-16 are consistent with a predominantly unordered structure. Addition of trifluoroethanol stabilized alpha-helical structure in the 1-34 amide and 7-34 amide peptides. The effect reached a plateau at a trifluoroethanol concentration of approximately 40%, and a helix content of some 23 residues was determined. PTHrP 1-34 amide interacted with palmitoyloleoylphosphatidyl serine vesicles and exhibited an increased alpha-helix content of approximately 12 helical residues. Similar results were observed for monomyristoyllecithin micelles and sodium dodecyl sulfate micelles. No interaction with dimyristoylphosphatidylcholine vesicles was detected by CD. The ability to bind to palmitoyloleoylphosphotidyl serine vesicles was also a feature of the 1-40 and 7-34 fragments, while the 1-16 fragment was apparently unaffected by interaction with this model membrane system. These results indicate that the conformational properties of the functionally significant amino terminal 1-34 region of PTHrP parallel those reported for the corresponding, but largely nonhomologous, region of parathyroid hormone. Conformational similarities may account for the ability of PTHrP to mimic the functional properties of parathyroid hormone.

Amino Acid Sequence↗

Conformation of parathyroid hormone: time-resolved fluorescence studies.

Conformational and environmental changes in the functionally significant amino-terminal region of human parathyroid hormone (hPTH), induced by solvent or by complexation with acidic lipid, have been investigated. Structural perturbations were monitored by their effect on the fluorescence decay kinetics of the single tryptophyl residue at position 23. Data for the intact hormone were compared with those for its 1-34 and 13-34 analogues. Deletion of the 35-84 sequence had no significant effect on the structure of hPTH in the region of Trp-23, nor was there any evidence for interaction of this region with the 1-12 sequence. On the basis of a comparison of the results of this study with structural information available from other spectroscopic techniques, we propose that the local structure in the region of Trp-23 of aqueous solutions of hPTH and hPTH 1-34 has helical character. This local structure was not stable in aqueous hPTH 13-34, but was present in hPTH and its analogues, both on complexation with acidic lipid and in helix-promoting solvents. The tryptophyl fluorescence of the lipid-bound peptides was characteristic of an aqueous environment. Triple-exponential fluorescence decay kinetics were observed for the tryptophyl residue of hPTH and its deletion analogues. This can be explained in terms of ground-state heterogeneity due to the presence of three C alpha-C beta rotamers of the tryptophanyl indole side chain. Assuming this model, we show that the calculated fractional concentrations of the decay time components correlate with the likely rotamer populations and with their expected dependence on the main-chain conformation.(ABSTRACT TRUNCATED AT 250 WORDS)

Circular Dichroism↗

Fluorescence decay kinetics of the tryptophyl residues of myoglobin: effect of heme ligation and evidence for discrete lifetime components.

The fluorescence decay kinetics of the tryptophyl residues of sperm whale and yellowfin tuna myoglobin have been determined by using time-correlated single photon counting, with picosecond resolution. Purification by HPLC techniques resulted in the isolation of samples that exclusively displayed picosecond decay kinetics. Lifetimes of 24.4 ps for Trp14 and 122.0 ps for Trp7 were found for oxy sperm whale myoglobin (pH 7), which agree with theoretical predictions [Hochstrasser, R. M., & Negus, D. K. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 4399-4403]. The effects of ligand binding and pH on the decay kinetics were investigated, and the results were shown to be consistent with the known crystal structures. Data for the met form of sperm whale myoglobin were analyzed both in terms of a sum of discrete exponential components and as a continuous gamma distribution of exponential decays. The results were not found to support the existence of multiple, structurally distinct conformation states in myoglobin.

Animals↗

Resolution of heterogeneous fluorescence into component decay-associated excitation spectra. Application to subtilisins.

Direct and indirect methods are described to combine steady-state and picosecond time-resolved fluorescence decay data to generate decay-associated excitation spectra. The heterogeneous fluorescence from a fluorophore mixture that models protein fluorescence was resolved into individual component excitation spectra. The two methods were also used to determine the excitation spectra associated with each of the decay time components for the proteins subtilisin Carlsberg and BPN'. On the basis of associated spectra, the decay components of both proteins were assigned to individual (or groups of) emitting species. The two approaches used to generate the decay-associated excitation spectra are compared and their general application to protein fluorescence studies is discussed.

Kinetics↗

The use of Stokes Raman scattering in time correlated single photon counting: application to the fluorescence lifetime of tyrosinate.

The decay kinetics of the tyrosinate form of tyrosine in aqueous solution, pH 11, were investigated by time correlated single photon counting using laser excitation. The lifetime of the single exponential decay was 30.5 +/- 0.2 ps. Stokes Raman scattering was used to determine the instrument response function and the advantages of this approach over the conventional Rayleigh scattering method are discussed.

Kinetics↗

A fluorogenic, mixed-disulfide reagent for thiol assay.

The design and synthesis of the mixed disulfide (ortho-nitroaniline-N-ethyldithio) 2-[pyridyl-5-thioureido-N'-(5-fluorescein)] are described and the chemical and spectroscopic properties of this thiol-specific fluorogenic reagent are presented. The high reactivity and sensitivity of this reagent in thiol assay are demonstrated with low-molecular-weight thiols and with human carbonmonoxyhemoglobin and its subunits. Comparison with conventional absorption methods shows that at least 100 times less material is needed; moreover, high background absorbance or turbidity do not interfere with the assay.

Carboxyhemoglobin↗

Recurrent breast cancer treated with the antioestrogen tamoxifen: correlation between hormonal changes and clinical course.

Forty-five post-menopausal women with recurrent breast cancer were treated with the antioestrogen, tamoxifen, 20 mg twice daily. Clinical assessment after 12 weeks indicated that 18 (40%) showed some remission. Gonadotrophins were suppressed within two weeks to relatively constant concentrations within the post-menopausal range, responses to luteinising hormone-releasing hormone (LH-RH) did not change, and androgen concentrations remained within the normal range in all patients. Oestradiol concentrations rose steadily only in women in whom treatment failed. Serum prolactin concentrations were raised in 18 out of the 44 (41%) patients in whom they were measured; 13 of these did not respond to treatment. Treatment did not change the average prolactin concentration when this was within the normal range, but it significantly reduced prolactin concentrations in hyperprolactinaemic patients--within two weeks (P less than 0-01) in those who responded well and by six weeks (P less than 0-05) in those who showed no remission. Among patients with normal prolactin values the release of prolactin after thyrotrophin-releasing hormone was significantly greater in those with no remission than in those who responded to tamoxifen. Responses in those with hyperprolactinaemia were reduced to about half the control values, and again this change occurred faster in those who were successfully treated. Patients therefore seem to have a better chance of responding to anti-oestrogen treatment if prolactin secretion is low.

Androgens↗

Hormonal effects of tamoxifen in oligospermic men.

The hormonal effects of tamoxifen (10 mg daily for 6 months) have been studied in nine men with oligospermia. Basal concentrations of serum LH (1-7 +/- 0-1 (S.E.M.) i.u./l) increased to a maximum of 4-1 +/- 1-3 i.u./l (P less than 0-001) after 6 months, and FSH rose from 4-9 +/- 1-0 to a maximum of 7-7 +/- 1-3 i.u./l after 4 months of treatment (P less than 0-01). The response to luteinizing hormone releasing hormone (LH-RH) was studied at monthly intervals. Sums of increments of serum LH increased from 35 +/- 4 to 92 +/- 17 i.u./l at 4 months (P less than 0-001) and of FSH from 14 +/- 3-4 to 23 +/- 3-5 i.u./l at 4 months (P less than 0-01). Basal serum androgens rose from 25 +/- 2-7 to 38 +/- 2-4 nmol/l after 4 months of treatment (P less than 0-05), and serum oestradiol-17beta increased from 185 +/- 25 to 631 +/- 90 pmol/l by 6 months (P less than 0-001). No significant changes occurred in sperm counts. Five normal men acted as controls: they were given tamoxifen for 1 week. No significant changes were observed in serum LH, FSH or release of these hormones following administration of LH-RH. Serum androgens and oestrogens however, increased significantly by day 4 of treatment (P less than 0-05).

Androgens↗