Out of the frying pan into the fire: Predicted warming in alpine streams suggests hidden consequences for aquatic ectotherms.

Thermal regimes of aquatic ecosystems are predicted to change as climate warming progresses over the next century, with high-latitude and high-elevation regions predicted to be particularly impacted. Here, we have modelled alpine stream water temperatures from air temperature data and used future predicted air temperature trajectories (representative concentration pathway [rcp] 4.5 and 8.5) to predict future water temperatures. Modelled stream water temperatures have been used to calculate cumulative degree days (CDDs) under current and future climate conditions. These calculations show that degree days will accumulate more rapidly under the future climate scenarios, and with a stronger effect for higher CDD values (e.g., rcp 4.5: 18-28 days earlier [CDD = 500]; 42-55 days earlier [CDD = 2000]). Changes to the time to achieve specific CDDs may have profound and unexpected consequences for alpine ecosystems. Our calculations show that while the effect of increased CDDs may be relatively small for organisms that emerge in spring-summer, the effects for organisms emerging in late summer-autumn may be substantial. For these organisms, the air temperatures experienced upon emergence could reach 9°C (rcp 4.5) or 12°C (rcp 8.5) higher than under current climate conditions, likely impacting on the metabolism of adults, the availability of resources, including food and suitable oviposition habitat, and reproductive success. Given that the movement of aquatic fauna to the terrestrial environment represents an important flux of energy and nutrients, differential changes in the time periods to achieve CDDs for aquatic and terrestrial fauna may de-couple existing predator-prey interactions.

Prediction and monitoring of relapse in stage III melanoma using circulating tumor DNA.

BACKGROUND: The advent of effective adjuvant therapies for patients with resected melanoma has highlighted the need to stratify patients based on risk of relapse given the cost and toxicities associated with treatment. Here we assessed circulating tumor DNA (ctDNA) to predict and monitor relapse in resected stage III melanoma. PATIENTS AND METHODS: Somatic mutations were identified in 99/133 (74%) patients through tumor tissue sequencing. Personalized droplet digital PCR (ddPCR) assays were used to detect known mutations in 315 prospectively collected plasma samples from mutation-positive patients. External validation was performed in a prospective independent cohort (n = 29). RESULTS: ctDNA was detected in 37 of 99 (37%) individuals. In 81 patients who did not receive adjuvant therapy, 90% of patients with ctDNA detected at baseline and 100% of patients with ctDNA detected at the postoperative timepoint relapsed at a median follow up of 20 months. ctDNA detection predicted patients at high risk of relapse at baseline [relapse-free survival (RFS) hazard ratio (HR) 2.9; 95% confidence interval (CI) 1.5-5.6; P = 0.002] and postoperatively (HR 10; 95% CI 4.3-24; P < 0.001). ctDNA detection at baseline [HR 2.9; 95% CI 1.3-5.7; P = 0.003 and postoperatively (HR 11; 95% CI 4.3-27; P < 0.001] was also associated with inferior distant metastasis-free survival (DMFS). These findings were validated in the independent cohort. ctDNA detection remained an independent predictor of RFS and DMFS in multivariate analyses after adjustment for disease stage and BRAF mutation status. CONCLUSION: Baseline and postoperative ctDNA detection in two independent prospective cohorts identified stage III melanoma patients at highest risk of relapse and has potential to inform adjuvant therapy decisions.

Surveillance imaging with FDG-PET/CT in the post-operative follow-up of stage 3 melanoma.

Background: As early detection of recurrent melanoma maximizes treatment options, patients usually undergo post-operative imaging surveillance, increasingly with FDG-PET/CT (PET). To assess this, we evaluated stage 3 melanoma patients who underwent prospectively applied and sub-stage-specific schedules of PET surveillance. Patients and methods: From 2009, patients with stage 3 melanoma routinely underwent PET +/- MRI brain scans via defined schedules based on sub-stage-specific relapse probabilities. Data were collected regarding patient characteristics and outcomes. Contingency analyses were carried out of imaging outcomes. Results: One hundred and seventy patients (stage 3A: 34; 3B: 93; 3C: 43) underwent radiological surveillance. Relapses were identified in 65 (38%) patients, of which 45 (69%) were asymptomatic. False-positive imaging findings occurred in 7%, and 6% had treatable second (non-melanoma) malignancies. Positive predictive values (PPV) of individual scans were 56%-83%. Negative scans had predictive values of 89%-96% for true non-recurrence [negative predictive values (NPV)] until the next scan. A negative PET at 18 months had NPVs of 80%-84% for true non-recurrence at any time in the 47-month (median) follow-up period. Sensitivity and specificity of the overall approach of sub-stage-specific PET surveillance were 70% and 87%, respectively. Of relapsed patients, 33 (52%) underwent potentially curative resection and 10 (16%) remained disease-free after 24 months (median). Conclusions: Application of sub-stage-specific PET in stage 3 melanoma enables asymptomatic detection of most recurrences, has high NPVs that may provide patient reassurance, and is associated with a high rate of detection of resectable and potentially curable disease at relapse.

Revision of the genus Caenota Mosely (Trichoptera: Calocidae), with descriptions of 2 new species and the larva of C. nemorosa Neboiss.

The caddisfly genus Caenota Mosely 1953 (in Mosely & Kimmins 1953) currently contains 5 species known from eastern Australia. Caenota is distinguished from other Calocidae genera by having adult males with greatly expanded maxillary palpi and a large membranous process associated with the antennal scape. Of the 5 described species, the larvae of only 1 is known. Here, we describe 2 new species, Caenota cudonis sp. nov. and C. equustagna sp. nov., from adult, larval, and pupal material. Also, we describe for the first time the larva of C. nemorosa Neboiss. These descriptions increase the number of Caenota species to 7 and the number of associated and described larvae to 4. This paper also provides descriptions of features associated with the adult head capsule of all described species of Caenota. Each of the known species is considered, with illustrations and re-descriptions of these features given.

Restoration of tumor suppression in prostate cancer by targeting the E3 ligase E6AP.

Restoration of tumor suppression is an attractive onco-therapeutic approach. It is particularly relevant when a tumor suppressor is excessively degraded by an overactive oncogenic E3 ligase. We previously discovered that the E6-associated protein (E6AP; as classified in the human papilloma virus context) is an E3 ligase that has an important role in the cellular stress response, and it directly targets the tumor-suppressor promyelocytic leukemia protein (PML) for proteasomal degradation. In this study, we have examined the role of the E6AP-PML axis in prostate cancer (PC). We show that knockdown (KD) of E6AP expression attenuates growth of PC cell lines in vitro. We validated this finding in vivo using cell line xenografts, patient-derived xenografts and mouse genetics. We found that KD of E6AP attenuates cancer cell growth by promoting cellular senescence in vivo, which correlates with restoration of tumor suppression by PML. In addition, we show that KD of E6AP sensitizes cells to radiation-induced death. Overall, our findings demonstrate a role for E6AP in the promotion of PC and support E6AP targeting as a novel approach for PC treatment, either alone or in combination with radiation.

Disposition of topical flurbiprofen in normal and aphakic rabbit eyes.

In a study of the ocular absorption and elimination of a topically applied non-steroidal anti-inflammatory drug (NSAID), flurbiprofen, the compound was well absorbed into rabbit ocular tissues and was highly concentrated in the rabbit cornea. In aphakic eyes, more drug penetrated to the vitreous and choroid-retina area than in normal rabbit eyes, although corneal concentrations were still high. No ocular metabolism of flurbiprofen could be detected, and the ocular route of application did not lead to any changes in blood elimination rates or metabolism when compared with intravenously injected drug. Currently, no NSAID is available for topical ocular use, and the development of such a drug is desirable for treatment of ocular inflammations, especially when long-term treatment is indicated.

Effect of benzalkonium chloride/EDTA on the ocular bioavailability of ketorolac tromethamine following ocular instillation to normal and de-epithelialized corneas of rabbits.

This study was designed to examine the effect of benzalkonium chloride/ethylenediaminetetraacetic acid (BAK/EDTA) on the ocular bioavailability (Focular) of ketorolac tromethamine after ocular instillation to normal and de-epithelialized corneas of rabbits both in vitro and in vivo. The in vitro Focular of the formulations was measured in flow-through perfusion chambers. For in vivo studies, a 35 microL dose of 0.5% ketorolac tromethamine with or without BAK/EDTA was instilled into rabbit eyes with intact or de-epithelialized corneas. At 0.5, 1, 2, 4, 6, and 8 h postdose, rabbits were euthanized, and the corneas and aqueous humor were collected from both eyes. The ketorolac concentrations from both in vivo and in vitro samples were quantified by reversed-phase high-performance liquid chromatography. The in vitro study results indicated that BAK/EDTA statistically significantly increased the Focular of ketorolac through de-epithelialized corneas but not through intact corneas. The in vivo study results showed that BAK/EDTA had no effect on the Focular of ketorolac in rabbits with intact corneas, based on the values of the area under the aqueous humor concentration versus time curves (AUC0-6h) of ketorolac. As expected, de-epithelialization of the corneas produced a faster and greater ocular absorption of ketorolac as evidenced by the smaller Tmax and larger AUC values compared to those for the intact corneas in vivo. However, BAK/EDTA decreased the ocular absorption of ketorolac in rabbits with de-epithelialized corneas. The half-lives (t 1/2) of ketorolac in corneal tissue and aqueous humor were longer in rabbits with intact corneas than those in rabbits with de-epithelialized corneas. In conclusion, the in vivo Focular of ketorolac was not altered by BAK/EDTA in rabbits with intact corneas, but it was decreased by BAK/EDTA in rabbits with de-epithelialized corneas. Therefore, the formulation with ketorolac alone may be better as a post-operative ocular analgesic.

Glucosinolate content and isothiocyanate evolution--two measures of the biofumigation potential of plants.

A total of 570 lyophilised Brassica root and shoot tissue samples were hydrolyzed, and the liberated isothiocyanates (ITCs) were analyzed by gas chromatography-flame photometric detection (GC-FPD). Glucosinolates (GSLs) were extracted from samples of the same tissues and analyzed by high-performance liquid chromatography (HPLC). The concentrations of six GSLs/ITCs (2-propenyl, 3-butyl, 4-pentenyl, benzyl, 4-methylthiobutyl, and 2-phenylethyl) as determined by the two techniques were compared. In 79% of the samples, the concentration of GSLs in the tissues was greater than that of the ITCs released on hydrolysis. Several possible reasons for the difference are proposed, including the effect of tissue storage time, hydrolysis of GSLs may be less efficient than the GSL extraction procedure, or some of the ITCs formed reacted with plant proteins and amino acids in the sample and were therefore not detected in the extract. GSL concentration in plant tissues is used to estimate the biofumigation potential of the plant tissue, whereas the actual biofumigation effect is thought to be due to the ITCs formed by hydrolysis of the plant-based GSLs. The variation between ITC and GSL values therefore has implications for the assessment of the biofumigation potential of the plant tissue.

Distribution of cyclosporin A in ocular tissues after topical administration to albino rabbits and beagle dogs.

PURPOSE: To determine the ocular pharmacokinetics of cyclosporin A after topical ophthalmic administration. METHODS: Radiolabled cyclosporin A in either a castor oil-in-water emulsion or a corn oil ointment was applied to the eyes of beagle dogs or albino rabbits using the following paradigms: (i) single doses of 0.2% emulsion to rabbits and dogs, (ii) single doses of 0.05%, 0.2%, or 0.4% emulsion to rabbits, (iii) multiple doses of 0.2% emulsion to dogs, (iv) single and multiple doses of 0.2% ointment to rabbits. The distribution of cyclosporin A was determined by measuring the distribution of radioactivity. RESULTS: After a single dose, cyclosporin A was rapidly absorbed into the conjunctiva (Cmax: dogs, 1490 ng/g; rabbits, 1340 ng/g) and cornea (Cmax: dogs, 311 ng/g; rabbits, 955 ng/g). High concentrations (>300 ng/g) could be detected in the cornea up to 96 hours post-dose. Lower concentrations were found in the intraocular tissues, and systemic absorption was minimal. After multiple doses, there was some accumulation in the cornea, lens, lacrimal gland, and iris-cilliary body, but limited accumulation in the conjunctiva and sclera. Ocular tissue concentrations of cyclosporin A increased with increasing dose concentration; proportionally in lacrimal gland and intraocular tissues; less than proportionally in conjunctiva and cornea. The pharmacokinetic profile of the cyclosporin A corn oil ointment was similar to that of the emulsion. CONCLUSIONS: Topical ophthalmic cyclosporin A penetrated into extraocular tissues at concentrations adequate for local immunomodulation while penetration into intraocular tissues was much less and absorption into the blood was minimal.

Comparison of concentration-time profiles of levobunolol and timolol in anterior and posterior ocular tissues of albino rabbits.

The potential effects of anti-glaucoma drugs, such as levobunolol and timolol, on blood flow in the posterior segment of the eye are of great interest in terms of changes in optic nerve head perfusion and prevention of visual field loss. These effects are related to the rate and extent of their absorption into the site of action. In this study, the concentrations of timolol and levobunolol in the aqueous humor, iris-ciliary body, vitreous humor, choroid-retina, and optic nerve were compared following instillation of a single drop of 0.5% ophthalmic solutions into albino rabbit eyes. Tissue drug and metabolite concentrations were measured by liquid chromatography-mass spectrometry. Dihydrobunolol (DHB) is an equipotent metabolite of levobunolol. In the anterior segment of the eye, levobunolol plus DHB concentrations were higher than timolol concentrations in aqueous humor and were comparable to those of timolol in iris-ciliary body. However, in the choroid-retina and optic nerve, timolol concentrations were greater than those of levobunolol plus DHB. Overall, the study demonstrates comparable concentrations of levobunolol and timolol in the anterior section of the eye. The low availability of levobunolol in the posterior segment as compared to timolol may be a key advantage for levobunolol in producing less adverse effect on blood flow in the choroid-retina and optic nerve.

Infantile hypertrophic pyloric stenosis: a Dunedin review.

Patients who had undergone a Ramstedt pyloromyotomy for infantile hypertrophic pyloric stenosis in Dunedin over a 10 year period to June 1984 were reviewed. Forty-two cases were recorded. More congenital anomalies than expected, a declining rate of wound complications and a high rate of a positive family history were found. The history is reviewed, the data presented and the results discussed.

Behaviour and fate of nine recycled water trace organics during managed aquifer recharge in an aerobic aquifer.

The fate of nine trace organic compounds was evaluated during a 12month large-scale laboratory column experiment. The columns were packed with aquifer sediment and evaluated under natural aerobic and artificial anaerobic geochemical conditions, to assess the potential for natural attenuation of these compounds during aquifer passage associated with managed aquifer recharge (MAR). The nine trace organic compounds were bisphenol A (BPA), 17ß-estradiol (E2), 17α-ethynylestradiol (EE2), N-nitrosodimethylamine (NDMA), N-nitrosomorpholine (NMOR), carbamazepine, oxazepam, iohexol and iodipamide. In the low organic carbon content Spearwood sediment, all trace organics were non-retarded with retardation coefficients between 1.0 and 1.2, indicating that these compounds would travel at near groundwater velocities within the aquifer. The natural aerobic geochemical conditions provided a suitable environment for the rapid degradation for BPA, E2, iohexol (half life <1day). Lag-times for the start of degradation of these compounds ranged from <15 to 30days. While iodipamide was persistent under aerobic conditions, artificial reductive geochemical conditions promoted via the addition of ethanol, resulted in rapid degradation (half life <1days). Pharmaceuticals (carbamazepine and oxazepam) and disinfection by-products (NDMA and NMOR) did not degrade under either aerobic or anaerobic aquifer geochemical conditions (half life >50days). Field-based validation experiments with carbamazepine and oxazepam also showed no degradation. If persistent trace organics are present in recycled waters at concentrations in excess of their intended use, natural attenuation during aquifer passage alone may not result in extracted water meeting regulatory requirements. Additional pre treatment of the recycled water would therefore be required.

Fate of nine recycled water trace organic contaminants and metal(loid)s during managed aquifer recharge into a anaerobic aquifer: Column studies.

Water quality changes associated with the passage of aerobic reverse osmosis (RO) treated recycled water through a deep anaerobic pyritic aquifer system was evaluated in sediment-filled laboratory columns as part of a managed aquifer recharge (MAR) strategy. The fate of nine recycled water trace organic compounds along with potential negative water quality changes such as the release of metal(loid)s were investigated in large-scale columns over a period of 12 months. The anaerobic geochemical conditions provided a suitable environment for denitrification, and rapid (half-life <1-25 days) degradation of the endocrine disrupting compounds (bisphenol A, 17beta-estradiol, 17alpha-ethynylestradiol), and iodipamide. However, pharmaceuticals (carbamazepine and oxazepam), disinfection by-products (N-nitrosodimethylamine, N-nitrosomorpholine) and iohexol did not degrade rapidly (half-life > 100 days). High retardation coefficients (R) determined for many of the trace organics (R 13 to 67) would increase aquifer residence time and be beneficial for many of the slow degrading compounds. However, for the trace organics with low R values (1.1-2.6) and slow degradation rates (half-life > 100 days), such as N-nitrosodimethylamine, N-nitrosomorpholine and iohexol, substantial biodegradation during aquifer passage may not occur and additional investigations are required. Only minor transient increases in some metal(loid) concentrations were observed, as a result of either pyrite oxidation, mineral dissolution or pH induced metal desorption, followed by metal re-sorption downgradient in the oxygen depleted zone.

Delineating the epithelial hierarchy in the mouse mammary gland.

Reconstitution assays have shown that mouse mammary stem cells reside within the mature mammary gland in vivo. Single cells could be prospectively isolated and shown to regenerate an entire mammary gland that exhibited full developmental capacity. The more recent identification of luminal progenitor populations has indicated that the mammary epithelium is organized in a hierarchical manner. Further definition of epithelial cell types in both mouse and human mammary glands will provide insight into the "cells of origin" in the different subtypes of breast cancer, as well as the nature of cancer-propagating cells. Here, we review the known characteristics of mammary stem and progenitor cells, their steroid receptor status, and the pathways that have thus far been implicated in regulating their self-renewal and differentiation.