Ice seals as sentinels for algal toxin presence in the Pacific Arctic and subarctic marine ecosystems

Domoic acid (DA) and saxitoxin (STX)-producing algae are present in Alaskan seas, presenting exposure risks to marine mammals that may be increasing due to climate change. To investigate potential increases in exposure risks to four pagophilic ice seal species (Erignathus barbatus, bearded seals; Pusa hispida, ringed seals; Phoca largha, spotted seals; and Histriophoca fasciata, ribbon seals), this study analyzed samples from 998 seals harvested for subsistence purposes in western and northern Alaska during 2005–2019 for DA and STX. Both toxins were detected in bearded, ringed, and spotted seals, though no clinical signs of acute neurotoxicity were reported in harvested seals. Bearded seals had the highest prevalence of each toxin, followed by ringed seals. Bearded seal stomach content samples from the Bering Sea showed a significant increase in DA prevalence with time (logistic regression, p = .004). These findings are consistent with predicted northward expansion of DA-producing algae. A comparison of paired samples taken from the stomachs and colons of 15 seals found that colon content consistently had higher concentrations of both toxins. Collectively, these results suggest that ice seals, particularly bearded seals (benthic foraging specialists), are suitable sentinels for monitoring HAB prevalence in the Pacific Arctic and subarctic.     

Organic matter influences phytotoxicity of cadmium to soybeans

Summary Soybeans (Glycine max L. var. Williams) were grown for six weeks in a greenhouse in quartz sand containing 0, 0.5, 1, 2, 4 or 8% (w/w) sterilized peat moss. The cation exchange capacities of the organic matter-sand (OM-S) mixtures ranged from 0.01 to 8.88 meq/100 g dry weight. Imposed on each OM-S mixture was a treatment of 0, 1.25, 2.5, 5.0, 10.0 or 20.0 ppm Cd applied as CdCl₂·21/2H₂O. Height growth was measured weekly and at harvest plants were separated into leaves, stems and roots for dry weight and tissue Cd determinations. For plants grown in sand alone, height growth and dry matter accumulation in all tissues were reduced and Cd content was increased. These effects were correlated with increasing Cd concentration in the rooting medium. Inhibitions in growth by Cd were reduced by addition of organic matter; the amount of alleviation was dependent on both the level of organic matter and the cadmium treatment. In the 0, 0.5 and 1% OM-S mixtures, Cd content in the various tissues was correlated with metal treatment. Tissue levels were markedly reduced for Cd treatments in the 2, 4 and 8% OM-S mixtures, although there was a positive correlation between tissue Cd and the 1.25 and 2.5 Cd treatments. The order of Cd accumulation in the tissues was roots stems>leaves.     

Effects of shelter and enrichment on the ecology and nutrient cycling of microbial communities of subtidal carbonate sediments

The interactions between physical disturbances and biogeochemical cycling are fundamental to ecology. The benthic microbial community controls the major pathway of nutrient recycling in most shallow-water ecosystems. This community is strongly influenced by physical forcing and nutrient inputs. Our study tests the hypotheses that benthic microbial communities respond to shelter and enrichment with (1) increased biomass, (2) change in community composition and (3) increased uptake of inorganic nutrients from the water column. Replicate in situ plots were sheltered from physical disturbance and enriched with inorganic nutrients or left without additional nutrients. At t(0) and after 10?days, sediment-water fluxes of nutrients, O(2) and N(2) , were measured, the community was characterized with biomarkers. Autochthonous benthic microalgal (BMA) biomass increased 30% with shelter and a natural fivefold increase in nutrient concentration; biomass did not increase with greater enrichment. Diatoms remained the dominant taxon of BMA, suggesting that the sediments were not N or Si limited. Bacteria and other heterotrophic organisms increased with enrichment and shelter. Daily exchanges of inorganic nutrients between sediments and the water column did not change in response to shelter or nutrient enrichment. In these sediments, physical disturbance, perhaps in conjunction with nutrient enrichment, was the primary determinant of microbial biomass.     

Protein misfolding and aggregation

Interest in the problem of protein misfolding and aggregation has exploded in recent years for two reasons: (1) the sharp rise in the number and volume of therapeutic proteins produced commercially and (2) the recognition of the central role of protein aggregates in degenerative diseases. The systematic study of protein aggregation presents major challenges to both the experimentalist and the theoretician. Much of the work retains an empirical flavor due to the experimental complexities; the sensitivity of protein aggregation to the slightest change in protein amino acid composition, solvent properties, or protein concentration; and the lack of robust theoretical models of misfolding and aggregation. Novel experimental and computational approaches are being developed, and we anticipate substantial progress will be made in the near future. Several presentations describing the latest advances in protein misfolding and aggregation were given at the American Chemical Society meeting (BIOT division) held in September, 2006 in San Francisco.     

The co-crystal structure of ubiquitin carboxy-terminal hydrolase L1 (UCHL1) with a tripeptide fluoromethyl ketone (Z-VAE(OMe)-FMK)

UCHL1 is a 223 amino acid member of the UCH family of deubiquitinating enzymes (DUBs), found abundantly and exclusively expressed in neurons and the testis in normal tissues. Two naturally occurring variants of UCHL1 are directly involved in Parkinson’s disease (PD). Not only has UCHL1 been linked to PD, but it has oncogenic properties, having been found abnormally expressed in lung, pancreatic, and colorectal cancers. Although inhibitors of UCHL1 have been described previously the co-crystal structure of the enzyme bound to any inhibitor has not been reported. Herein, we report the X-ray structure of UCHL1 co-crystallized with a peptide-based fluoromethylketone inhibitor, Z-VAE(OMe)-FMK (VAEFMK) at 2.35 Å resolution. The co-crystal structure reveals that the inhibitor binds in the active-site cleft, irreversibly modifying the active-site cysteine; however, the catalytic histidine is still misaligned as seen in the native structure, suggesting that the inhibitor binds to an inactive form of the enzyme. Our structure also reveals that the inhibitor approaches the active-site cleft from the opposite side of the crossover loop as compared to the direction of approach of ubiquitin’s C-terminal tail, thereby occupying the P1′ (leaving group) site, a binding site perhaps used by the unknown C-terminal extension of ubiquitin in the actual in vivo substrate(s) of UCHL1. This structure provides a view of molecular contacts at the active-site cleft between the inhibitor and the enzyme as well as furnishing structural information needed to facilitate further design of inhibitors targeted to UCHL1 with high selectivity and potency.     

Proteomic analysis of the venom of Heterometrus longimanus (Asian black scorpion)

Venoms have evolved over millions of years into potent cocktails of bioactive peptides and proteins. These compounds can be of great value to the pharmaceutical industry for numerous clinical applications. In this study, a novel proteomic - bioinformatic approach was utilised, where chromatography followed by gel electrophoresis was utilised to separate the venom peptides/proteins of Heterometrus longimanus (Asian black scorpion). Purified peptides were analysed by tandem mass spectrometry, de novo sequenced and then homology matched against known peptides in the Swiss-Prot protein database. Numerous potentially biologically active peptide matches were discovered, and a simple scoring system applied to putatively assign functions to the peptides. As a validation of this approach, the functional composition of the experimentally derived proteome is similar to that of other scorpions, and contains a potent mix of toxins, antimicrobials and ionic channel inhibitors.     

General practitioner attitudes towards referral of eating-disordered patients: a vignette study based on the theory of planned behaviour

Objective The study examined individual differences between general practitioners (GPs) to determine their impact on variations in intention to refer a hypothetical patient with disordered eating to specialist eating disorder services. The study also examined the impact of patient weight on intention to refer.Method GPs within three primary care trusts (PCTs) were posted a vignette depicting a patient with disordered eating, described as either normal weight or underweight. A questionnaire was developed from the theory of planned behaviour to assess the GPs' attitudes, perception of subjective norms, perceived behavioural control, and intention to refer the patient. Demographic details were also collected.Results Responses were received from 88 GPs (33%). Intention to refer the patient was significantly related to subjective norms and cognitive attitudes. Together these predictors explained 86% of the variance in the intention to refer. GP or practice characteristics did not have a significant effect on the GPs' intention to refer, and nor did the patient's weight.Conclusion Despite National Institute for Health and Clinical Excellence current guidance, patient weight did not influence GPs' decisions to refer. Much of the variance in actual referral behaviour may be explained by cognitive attitudes and subjective norms. Interventions to reduce this variation should be focused on informing GPs about actual norms, and best practice guidelines.     

SPECIAL PAPER: THE PLASTOCHRON INDEX: A REVIEW AFTER TWO DECADES OF USE

The plastochron index provides a morphological time scale which has proved more reliable than chronological age in studies relating morphological and physiological development of a whole plant or plant organ. Since its inception in 1957, the index has been utilized in a variety of investigations from leaf ontogeny in cottonwood trees to rhizoid cluster initiation in algae. The plastochron index has been extensively used in studies involving source and sink relationships, leaf anatomy, cell differentiation, and primary vascularization. It has been used in investigations of hormonal regulation of plant growth and in studies of the effects of various environmental factors on developmental processes in crops. This paper reviews some of the literature from 1957 to present concerning the development and use of the plastochron index.     

Translesion synthesis past platinum DNA adducts by human DNA polymerase mu

DNA polymerase mu (pol mu) is a member of the pol X family of DNA polymerases, and it shares a number of characteristics of both DNA polymerase beta (pol beta) and terminal deoxynucleotidyl transferase (TdT). Because pol beta has been shown to perform translesion DNA synthesis past cisplatin (CP)- and oxaliplatin (OX)-GG adducts, we determined the ability of pol mu to bypass these lesions. Pol mu bypassed CP and OX adducts with an efficiency of 14-35% compared to chain elongation on undamaged DNA, which is second only to pol eta in terms of bypass efficiency. The relative ability of pol mu to bypass CP and OX adducts was dependent on both template structure and sequence context. Since pol mu has been shown to be more efficient on gapped DNA templates than on primed single-stranded DNA templates, we determined the ability of pol mu to bypass Pt-DNA adducts on both primed single-stranded and gapped templates. The bypass of Pt-DNA adducts by pol mu was highly error-prone on all templates, resulting in 2, 3, and 4 nt deletions. We postulate that bypass of Pt-DNA adducts by pol mu may involve looping out the Pt-GG adduct to allow chain elongation downstream of the adduct. This reaction appears to be facilitated by the presence of a downstream "acceptor" and a gap large enough to provide undamaged template DNA for elongation past the adduct, although gapped DNA is clearly not required for bypass.