Identifying behavioral states and habitat use of acoustically tracked humpback whales in Hawaii

Although humpback whales have been well‐studied on their Hawaiian breeding grounds, it is difficult to track individual animals over long distances without tags, particularly when they move offshore. Here, singing humpback whales were localized in three dimensions on the Pacific Missile Range Facility off Kauai, Hawaii, located between 20 km and 80 km offshore, from January 2011 through June 2014. Detailed behavioral analyses were conducted on the resulting tracks. One hundred and eight individual tracks were identified and metrics of these tracks were examined. Using these metrics, the tracks were classified into four behavior categories, described herein as Directed Travel, Repeated Stationary Dives, Mill, and tracks with Combinations of behavioral states. Some diel and seasonal patterns were identified, with Mill tracks occurring more at night than the other behaviors, Repeated Stationary Dive tracks occurring more during the day, and Directed Travel occurring only at the start and end of the breeding season. These results provide detailed insights into the movement of singing humpback whales, particularly in offshore waters where they may be migrating into or out of breeding grounds. This also contributes valuable information on the baseline behavior of humpback whales on a US Navy training range.     

TROSY NMR with a 52 kDa sugar transport protein and the binding of a small-molecule inhibitor

Using the sugar transport protein, GalP, from Escherichia coli, which is a homologue of human GLUT transporters, we have overcome the challenges for achieving high-resolution [¹⁵N-¹H]- and [¹³C-¹H]-methyl-TROSY NMR spectra with a 52?kDa membrane protein that putatively has 12 transmembrane-spanning α-helices and used the spectra to detect inhibitor binding. The protein reconstituted in DDM detergent micelles retained structural and functional integrity for at least 48?h at a temperature of 25?°C as demonstrated by circular dichroism spectroscopy and fluorescence measurements of ligand binding, respectively. Selective labelling of tryptophan residues reproducibly gave 12 resolved signals for tryptophan ¹⁵N backbone positions and also resolved signals for ¹⁵N side-chain positions. For improved sensitivity isoleucine, leucine and valine (ILV) methyl-labelled protein was prepared, which produced unexpectedly well resolved [¹³C-¹H]-methyl-TROSY spectra showing clear signals for the majority of methyl groups. The GalP/GLUT inhibitor forskolin was added to the ILV-labelled sample inducing a pronounced chemical shift change in one Ile residue and more subtle changes in other methyl groups. This work demonstrates that high-resolution TROSY NMR spectra can be achieved with large complex α-helical membrane proteins without the use of elevated temperatures. This is a prerequisite to applying further labelling strategies and NMR experiments for measurement of dynamics, structure elucidation and use of the spectra to screen ligand binding.     

Receptor Activated Ca2+ Release Is Inhibited by Boric Acid in Prostate Cancer Cells

Background

The global disparity in cancer incidence remains a major public health problem. We focused on prostate cancer since microscopic disease in men is common, but the incidence of clinical disease varies more than 100 fold worldwide. Ca²⁺ signaling is a central regulator of cell proliferation, but has received little attention in cancer prevention. We and others have reported a strong dose-dependent reduction in the incidence of prostate and lung cancer within populations exposed to boron (B) in drinking water and food; and in tumor and cell proliferation in animal and cell culture models.

Methods/Principal Findings

We examined the impact of B on Ca²⁺ stores using cancer and non-cancer human prostate cell lines, Ca²⁺ indicators Rhod-2 AM and Indo-1 AM and confocal microscopy. In DU-145 cells, inhibition of Ca²⁺ release was apparent following treatment with Ringers containing RyR agonists cADPR, 4CmC or caffeine and respective levels of BA (50 µM), (1, 10 µM) or (10, 20, 50,150 µM). Less aggressive LNCaP cancer cells required 20 µM BA and the non-tumor cell line PWR1E required 150 µM BA to significantly inhibit caffeine stimulated Ca²⁺ release. BA (10 µM) and the RyR antagonist dantroline (10 µM) were equivalent in their ability to inhibit ER Ca²⁺ loss. Flow cytometry and confocal microscopy analysis showed exposure of DU-145 cells to 50 µM BA for 1 hr decreased stored [Ca²⁺] by 32%.

Conclusion/Significance

We show B causes a dose dependent decrease of Ca²⁺ release from ryanodine receptor sensitive stores. This occurred at BA concentrations present in blood of geographically disparate populations. Our results suggest higher BA blood levels lower the risk of prostate cancer by reducing intracellular Ca²⁺ signals and storage.     

Characterization of Two Distinct Lymphoproliferative Diseases Caused by Ectopic Expression of the Notch Ligand DLL4 on T Cells

Notch signaling is essential for the development of T cell progenitors through the interaction of NOTCH1 receptor on their surface with the ligand, Delta-like 4 (DLL4), which is expressed by the thymic epithelial cells. Notch signaling is quickly shut down once the cells pass β-selection, and CD4/CD8 double positive (DP) cells are unresponsive to Notch. Over the past two decades a number of papers reported that over-activation of Notch signaling causes T cell acute lymphoblastic leukemia (T-ALL), a cancer that prominently features circulating monoclonal CD4/CD8 double positive T cells in different mouse models. However, the possible outcomes of Notch over-activation at different stages of T cell development are unknown, and the fine timing of Notch signaling that results in T-ALL is poorly understood. Here we report, by using a murine model that ectopically expresses DLL4 on developing T cells, that the T-ALL onset is highly dependent on a sustained Notch activity throughout the DP stage, which induces additional mutations to further boost the signaling. In contrast, a shorter period of Notch activation that terminates at the DP stage causes a polyclonal, non-transmissible lymphoproliferative disorder that is also lethal. These observations resolved the discrepancy of previous papers on DLL4 driven hematological diseases in mice, and show the critical importance of the timing and duration of Notch activity.     

Dependence of proteasome processing rate on substrate unfolding

Protein degradation by eukaryotic proteasomes is a multi-step process involving substrate recognition, ATP-dependent unfolding, translocation into the proteolytic core particle, and finally proteolysis. To date, most investigations of proteasome function have focused on the first and the last steps in this process. Here we examine the relationship between the stability of a folded protein domain and its degradation rate. Test proteins were targeted to the proteasome independently of ubiquitination by directly tethering them to the protease. Degradation kinetics were compared for test protein pairs whose stability was altered by either point mutation or ligand binding, but were otherwise identical. In both intact cells and in reactions using purified proteasomes and substrates, increased substrate stability led to an increase in substrate turnover time. The steady-state time for degradation ranged from ～5 min (dihydrofolate reductase) to 40 min (I27 domain of titin). ATP turnover was 110/min./proteasome, and was not markedly changed by substrate. Proteasomes engage tightly folded substrates in multiple iterative rounds of ATP hydrolysis, a process that can be rate-limiting for degradation.     

Are environmental factors important facilitators of pompom weed (<Emphasis Type="Italic">Campuloclinium macrocephalum</Emphasis>) invasion in South African rangelands?

The absence of natural enemies being keystone to the success of invasive alien plants (IAPs) can only be accepted once all the factors governing the invader and the ecosystems it invades have been established. Few studies have attempted this approach. This study reports on the relations between the invasive alien forb Campuloclinium macrocephalum (Less.) DC. (pompom weed), herbivory and the ecology of the rangelands invaded in South Africa. Eighty invaded rangelands in Gauteng Province were studied. Pompom weed herbivory was insignificant with the greatest damage being caused by native grasshoppers. Rainfall, topography and soil texture were the primary drivers of vegetation pattern. Pompom weed exhibited disturbance-mediated invasion strategies, favouring rangelands affected by non-sustainable commercial grazing practices, abandoned agricultural fields and drained wetlands. It invaded vegetation in poor condition with sward basal cover <19%. Weed density was exacerbated by high fire frequency. Vegetation susceptible to pompom weed was also invaded by other alien and native pasture weeds. The absence of natural enemies could give pompom weed a competitive advantage over native pioneer cohorts in disturbed rangelands, but is unlikely to increase fitness enabling its establishment in vegetation in good condition. Grazing strategies that promote the dominance of grass species adapted to frequent non-selective defoliation appear to restrict pompom weed better than degraded rangelands that are not utilised. Biotic resistance of rangelands to pompom weed invasion will be greatest in vegetation in good condition and under good management. The low abundance of pompom weed in rangelands in communal areas and fence-line contrasts in invaded and un-invaded road reserves warrants further investigation.     

Diet of two commercially important shark species in the United Arab Emirates: milk shark,Rhizoprionodon acutus (Rüppell, 1837), and slit‐eye shark,Loxodon macrorhinus (Müller & Henle, 1839)

The diets of the milk shark, Rhizoprionodon acutus, and the slit‐eye shark, Loxodon macrorhinus, landed from the artisanal fishery in the Arabian Gulf waters of the United Arab Emirates were investigated to determine their dietary preferences. Stomach contents from 57 milk sharks and 53 slit eye sharks were collected from Abu Dhabi (R. acutus, n = 23), Dubai (R. acutus, n = 5; L. macrorhinus, n = 15) and Ras Al Khaimah (R. acutus, n = 29; L. macrorhinus, n = 38) during fishery surveys from January to May 2012. Prey items were identified to the lowest possible taxonomic level, grouped into five categories including ‘teleost fish’, ‘cephalopods’, ‘crustaceans’, ‘invertebrates’, and ‘other’. The diets of both species were described using the numeric, frequency and weight methods, and the index of relative importance (IRI). The majority of stomachs for both species had food, with 66.6% of milk shark stomachs and 90.5% of slit‐eye shark stomachs containing prey items, both dominated by small teleosts. Rhizoprionodon acutus fed on a wide variety of teleost species, primarily Engraulidae (anchovies) (28%), Gerreidae (mojarras) (5.6%) and Carangidae (jacks) (1.6%) with occasional crustacean and cephalopod prey (8%). On the other hand, L. macrorhinus seemed to have a preference for one species in terms of teleosts (anchovies) (35.1%) and fed on a wider variety of crustaceans and cephalopods (22.6%). There was little overlap in the diets of these two species, suggesting that they may either be using different habitats or that in these waters, the milk shark is a generalist species while the slit‐eye is a specialist feeder.