Analysis of the water-soluble products of phosphatidylcholine breakdown by ion-exchange chromatography. Bombesin and TPA (12-O-tetradecanoylphorbol 13-acetate) stimulate choline generation in Swiss 3T3 cells by a common mechanism.

A method for the rapid and quantitative separation of glycerophosphocholine, choline phosphate and choline upon ion-exchange columns is described. The method has been utilized to examine the stimulation of phosphatidylcholine breakdown in quiescent Swiss 3T3 cells in response to bombesin and 12-O-tetradecanoylphorbol 13-acetate (TPA). The stimulated generation of choline is shown to precede that of choline phosphate, with no effect upon glycerophosphocholine levels; but was attenuated in cells in which protein kinase C activity was down-regulated. The results thus suggest that stimulation of the cells with either bombesin or TPA activates phospholipase D-catalysed phosphatidylcholine breakdown by a common mechanism involving the activation of protein kinase C.     

A quantitative model of DNA fragments generated by ionizing radiation, and possible experimental applications

We derive an equation for observed frequencies of DNA fragments as a function of size. In this derivation, we consider an experimental system where fragments are generated by random, independent double-strand breaks on chromosomes (or other large DNA molecules) and then separated by size on agarose gels. When visualizing these fragments using Southern hybridization techniques (employing a site-specific probe), we predict an intensity distribution that has unusual properties. In particular, peaks in the fragment size distribution depend not only on standard breakage parameters, but also on the location of the hybridization site. Our model is consistent with experimental and theoretical results reported elsewhere, where measurements of peaks are used for the physical mapping of genes. Further, we propose that similar experiments might be suitable for precise measurements of the parameters of double-strand breakage (as an alternative to neutral filter elutions and neutral sucrose gradients) and for testing the assumption of random, independent breakage for different types of radiation.     

ATP1A3 Mutation in Adult Rapid-Onset Ataxia

A 21-year old male presented with ataxia and dysarthria that had appeared over a period of months. Exome sequencing identified a de novo missense variant in ATP1A3, the gene encoding the α3 subunit of Na,K-ATPase. Several lines of evidence suggest that the variant is causative. ATP1A3 mutations can cause rapid-onset dystonia-parkinsonism (RDP) with a similar age and speed of onset, as well as severe diseases of infancy. The patient’s ATP1A3 p.Gly316Ser mutation was validated in the laboratory by the impaired ability of the expressed protein to support the growth of cultured cells. In a crystal structure of Na,K-ATPase, the mutated amino acid was directly apposed to a different amino acid mutated in RDP. Clinical evaluation showed that the patient had many characteristics of RDP, however he had minimal fixed dystonia, a defining symptom of RDP. Successive magnetic resonance imaging (MRI) revealed progressive cerebellar atrophy, explaining the ataxia. The absence of dystonia in the presence of other RDP symptoms corroborates other evidence that the cerebellum contributes importantly to dystonia pathophysiology. We discuss the possibility that a second de novo variant, in ubiquilin 4 (UBQLN4), a ubiquitin pathway component, contributed to the cerebellar neurodegenerative phenotype and differentiated the disease from other manifestations of ATP1A3 mutations. We also show that a homozygous variant in GPRIN1 (G protein-regulated inducer of neurite outgrowth 1) deletes a motif with multiple copies and is unlikely to be causative.     

Validation of yellowedge grouper, <Emphasis Type="Italic">Epinephelus flavolimbatus</Emphasis>, age using nuclear bomb-produced radiocarbon

Age validation and estimates of longevity of yellowedge grouper (Epinephelus flavolimbatus) from the Gulf of Mexico (GOM) are needed to inform fishery management decisions. Yellowedge grouper sagittal otoliths (n = 100) were collected, aged using conventional means, and cores were submitted for radiocarbon (¹⁴C) measurement. Radiocarbon values of yellowedge grouper otoliths were compared to established radiocarbon chronologies in the region to validate the age and ageing methodology of this species. The yellowedge grouper chronology displayed a similar sigmoidal trend as previously published chronologies. In addition to the core analysis, multiple areas on otolith sections from eight specimens were analyzed for Δ¹⁴C to validate age estimates for fish born prior to the ¹⁴C increase. Our results indicate that yellowedge grouper live longer than previously reported (minimum of 40 years based on radiocarbon measurements). The validated ageing methodology supported an estimated maximum longevity of 85 years and established that yellowedge grouper have the longest lifespan currently known for any species of grouper in the GOM. Results also indicate a depth-age interaction in that material extracted from adult otolith sections assigned to post-bomb dates exhibited lower Δ¹⁴C values than cores (juvenile material) assigned to the same post-bomb dates. This finding is likely explained by lower ¹⁴C levels reported from water masses at deeper depths (>100 m) which are inhabited by adults.     

A Passive Heat Maintenance Strategy Implemented during a Simulated Half-Time Improves Lower Body Power Output and Repeated Sprint Ability in Professional Rugby Union Players

Reduced physical performance has been observed following the half-time period in team sports players, likely due to a decrease in muscle temperature during this period. We examined the effects of a passive heat maintenance strategy employed between successive exercise bouts on core temperature (T_core) and subsequent exercise performance. Eighteen professional Rugby Union players completed this randomised and counter-balanced study. After a standardised warm-up (WU) and 15 min of rest, players completed a repeated sprint test (RSSA 1) and countermovement jumps (CMJ). Thereafter, in normal training attire (Control) or a survival jacket (Passive), players rested for a further 15 min (simulating a typical half-time) before performing a second RSSA (RSSA 2) and CMJ’s. Measurements of T_core were taken at baseline, post-WU, pre-RSSA 1, post-RSSA 1 and pre-RSSA 2. Peak power output (PPO) and repeated sprint ability was assessed before and after the simulated half-time. Similar T_core responses were observed between conditions at baseline (Control: 37.06±0.05°C; Passive: 37.03±0.05°C) and for all other T_core measurements taken before half-time. After the simulated half-time, the decline in T_core was lower (-0.74±0.08% vs. -1.54±0.06%, p<0.001) and PPO was higher (5610±105 W vs. 5440±105 W, p<0.001) in the Passive versus Control condition. The decline in PPO over half-time was related to the decline in T_core (r = 0.632, p = 0.005). In RSSA 2, best, mean and total sprint times were 1.39±0.17% (p<0.001), 0.55±0.06% (p<0.001) and 0.55±0.06% (p<0.001) faster for Passive versus Control. Passive heat maintenance reduced declines in T_core that were observed during a simulated half-time period and improved subsequent PPO and repeated sprint ability in professional Rugby Union players.     

Variation in vegetative water use in the savannas of the North Australian Tropical Transect

Abstract. The decline in tree density on sandy soils in savannas is highly correlated with declining mean annual rainfall along the North Australian Tropical Transect (NATT). We reanalyse various data on water use by individual trees and argue that a common relationship can be used to estimate annual water use by tree stands along the NATT from ca. 600 mm mean annual rainfall to at least 1600 mm. Where rainfall is less than 600 mm, trees of a given size use less water than at sites where rainfall is higher. We use these relationships to relate water use at the stand scale with mean annual rainfall along the NATT. From this we show that the empirical data imply that the minimum depth of sandy soil that needs to be exploited by trees declines with increasing aridity along the NATT from more than 5 m to less than 1 m. This finding is consistent with other observations and the pattern that with increasing aridity, an increasing proportion of rainfall coming from isolated storms rather than from periods of extended monsoon activity.     

A Theoretical Model for Utilizing Mammalian Pharmacology and Safety Data to Prioritize Potential Impacts of Human Pharmaceuticals to Fish

Due to the potential for long-term, low-level exposure of environmental species to pharmaceuticals in the environment, concerns over chronic ecotoxicity have been raised. Pharmaceuticals typically have specific enzyme and receptor-based modes of action, which are extensively studied in mammals during drug development. A survey of the literature demonstrated that there is conservation of many enzyme/receptor systems between mammalian and teleost systems. Based on this conservation of enzyme/receptor systems across teleost species, a model has been developed to utilize the information from mammalian pharmacology and toxicology studies to evaluate the potential for chronic receptor mediated responses in fish. In this model, a measured human therapeutic plasma concentration (H_TPC) is compared to a predicted steady state plasma concentration (F_SSPC) in fish, and an effect ratio (ER = H_TPC/F_SSPC) is computed. The lower the ER, the greater the potential for a pharmacological response in fish. Data collection and model validation will strengthen the applicability of this approach as a viable tool for prioritizing research initiatives that examine the potential impact of pharmaceuticals on fish.     

Coral calcification rates by the buoyant weight technique: Effects of alizarin staining

The buoyant weight method has been used in a laboratory experiment over an 18-day period to assess effects of alizarin staining on the calcification rate of the hermatypic coral Diploria strigosa (Dana). Exposure of corals to a concentration of 10 mg/l alizarin for 24 h in a flow-through system caused a significant depression in calcification for a period of up to 6 days. It is suggested that such initial calcification depressions after staining could affect absolute growth measurements and cause synergistic effects with experimental manipulation of environmental conditions unless an appropriate recovery period is allowed.