Rapid and simple isolation of pure photosystem II core and reaction center particles from spinach

Pure and active oxygen-evolving PS II core particles containing 35 Chl per reaction center were isolated with 75% yield from spinach PS II membrane fragments by incubation with n-dodecyl--D-maltoside and a rapid one step anion-exchange separation. By Triton X-100 treatment on the column these particles could be converted with 55% yield to pure and active PS II reaction center particles, which contained 6 Chl per reaction center.Abbreviations Bis-Tris bis[2-hydroxyethyl]imino-tris[hydroxymethyl]methane - Chl chlorophyll - CP29 Chl a/b protein of 29 kDa - Cyt b ₅₅₉ cytochrome b ₅₅₉ - DCBQ 2,5-dichloro-p-benzo-quinone - LHC II light-harvesting complex II, predominant Chl a/b protein - MES 2-[N-Morpholino]ethanesulfonic acid - Pheo pheophytin - PS H photosystem II - Q_A bound plastoquinone, serving as the secondary electron acceptor in PS II (after Pheo) - SDS sodiumdodecylsulfate     

Seabirds and marine mammals in the eastern Barents Sea: late summer at-sea distribution and calculated food intake

The at-sea distribution of seabirds and marine mammals in the eastern Barents Sea was determined using standardized transect counts during three cruises of RV“Dalnie Zelentsy” (Murmansk) in late summer 1991, 1992 and 1993. Totals of 32,268 seabirds, 485 pinnipeds, 25 cetaceans and 4 polar bears were counted during 554 half-hour counts. Numbers were converted into densities, total biomass and calculated daily food intake. Mean total food intake in kg fresh weight/km².day was 3.1 for the entire zone and all years; fish eaters dominated the whole region, with an intake of 1.3 (mainly Brünnich’s guillemot, Uria lomvia, and harp seal, Phoca groenlandica), followed by zooplankton eaters (0.85, mainly fulmar, Fulmarus glacialis) and mixed zooplankton and fish feeders (0.75, mainly minke whale, Balaenoptera acutorostrata, and kittiwake, Rissa tridactyla). Year-to-year variations were of little importance, while geographic differences were obvious between Norwegian coastal, Atlantic and Barents Sea water masses, both quantitatively and qualitatively (relative importance of main diets). Within each zone, a strong geographic heterogeneity was noted, with high local concentrations at fronts between water masses and at ice edges.     

Division of labor within the DNA damage tolerance system reveals non-epistatic and clinically actionable targets for precision cancer medicine

Crosslink repair depends on the Fanconi anemia pathway and translesion synthesis polymerases that replicate over unhooked crosslinks. Translesion synthesis is regulated via ubiquitination of PCNA, and independently via translesion synthesis polymerase REV1. The division of labor between PCNA-ubiquitination and REV1 in interstrand crosslink repair is unclear. Inhibition of either of these pathways has been proposed as a strategy to increase cytotoxicity of platinating agents in cancer treatment. Here, we defined the importance of PCNA-ubiquitination and REV1 for DNA in mammalian ICL repair. In mice, loss of PCNA-ubiquitination, but not REV1, resulted in germ cell defects and hypersensitivity to cisplatin. Loss of PCNA-ubiquitination, but not REV1 sensitized mammalian cancer cell lines to cisplatin. We identify polymerase Kappa as essential in tolerating DNA damage-induced lesions, in particular cisplatin lesions. Polk-deficient tumors were controlled by cisplatin treatment and it significantly delayed tumor outgrowth and increased overall survival of tumor bearing mice. Our results indicate that PCNA-ubiquitination and REV1 play distinct roles in DNA damage tolerance. Moreover, our results highlight POLK as a critical TLS polymerase in tolerating multiple genotoxic lesions, including cisplatin lesions. The relative frequent loss of Polk in cancers indicates an exploitable vulnerability for precision cancer medicine.     

Developing outcome measures for pediatric mitochondrial disorders: Which complaints and limitations are most burdensome to patients and their parents?

Since some drug intervention effects are only experienced by the patient, organizations such as the Food and Drug Administration prefer clinically meaningful outcome measures. Here, we evaluated which symptoms and limitations in daily life are most burdensome to pediatric patients with mitochondrial disorders and their parents, using two questionnaires. In a study of 78 patients, the most burdensome complaints included fatigue, behavior and speech disturbances, epilepsy and muscle weakness and a high degree of limitations in daily activities was found. Importantly, there was a discrepancy between what symptoms metabolic pediatricians estimated would be most burdensome compared to the patients'/caretakers' opinion. To include feasible and relevant outcome measures in intervention studies, the experience and opinions of patients and caretakers should therefore be heard.     

Protein degradation sets the fraction of active ribosomes at vanishing growth

Growing cells adopt common basic strategies to achieve optimal resource allocation under limited resource availability. Our current understanding of such “growth laws” neglects degradation, assuming that it occurs slowly compared to the cell cycle duration. Here we argue that this assumption cannot hold at slow growth, leading to important consequences. We propose a simple framework showing that at slow growth protein degradation is balanced by a fraction of “maintenance” ribosomes. Consequently, active ribosomes do not drop to zero at vanishing growth, but as growth rate diminishes, an increasing fraction of active ribosomes performs maintenance. Through a detailed analysis of compiled data, we show that the predictions of this model agree with data from E. coli and S. cerevisiae. Intriguingly, we also find that protein degradation increases at slow growth, which we interpret as a consequence of active waste management and/or recycling. Our results highlight protein turnover as an underrated factor for our understanding of growth laws across kingdoms.     

Estimation of the Use of Antibiotics in the Small Ruminant Industry in the Netherlands in 2011 and 2012

The aim of this study was to estimate the quantity of antibiotics and classes of antibiotics used in the small ruminant industry in the Netherlands in 2011 and 2012. Twelve large veterinary practices, located throughout the Netherlands were selected for this study. All small ruminant farms associated with these practices that had complete records on the quantity of antibiotics prescribed were included. The veterinary practices provided data on all antibiotics prescribed, and the estimated animal used daily dose of antibiotics per year (AUDD/Y) was calculated for each farm. The median AUDD/Y in small ruminant farms was zero in both years (mean 0.60 in 2011, and 0.62 in 2012). The largest quantity of antibiotic use was observed in the professional goat industry (herds of ≥32 goats) with a median AUDD/Y of 1.22 in 2011 and 0.73 in 2012. In the professional sheep industry (flocks of ≥32 sheep), the median AUDD/Y was 0 in 2011 and 0.10 in 2012. In the small scale industry (flocks or herds of <32 sheep or goats), the median AUDD/Y never exceeded 0. The most frequently prescribed antibiotics in the small scale industry and professional sheep farms belonged to the penicillin class. In professional goat farms, antibiotics of the aminoglycoside class were most frequently prescribed. This study provides the first assessment on the quantity of antibiotic use in the small ruminant industry. Given a comparable attitude towards antibiotic use, these results might be valid for small ruminant populations in other north-western European countries as well. The antibiotic use in the small ruminant industry appeared to be low, and is expected to play a minor role in the development of antibiotic resistance. Nevertheless, several major zoonotic bacterial pathogens are associated with the small ruminant industry, and it remains important that antibiotics are used in a prudent way.     

Desert breath—How fog promotes a novel type of soil biocenosis,forming the coastal Atacama Desert’s living skin

The Atacama Desert is the driest non‐polar desert on Earth, presenting precarious conditions for biological activity. In the arid coastal belt, life is restricted to areas with fog events that cause almost daily wet–dry cycles. In such an area, we discovered a hitherto unknown and unique ground covering biocenosis dominated by lichens, fungi, and algae attached to grit‐sized (~6 mm) quartz and granitoid stones. Comparable biocenosis forming a kind of a layer on top of soil and rock surfaces in general is summarized as cryptogamic ground covers (CGC) in literature. In contrast to known CGC from arid environments to which frequent cyclic wetting events are lethal, in the Atacama Desert every fog event is answered by photosynthetic activity of the soil community and thus considered as the desert's breath. Photosynthesis of the new CGC type is activated by the lowest amount of water known for such a community worldwide thus enabling the unique biocenosis to fulfill a variety of ecosystem services. In a considerable portion of the coastal Atacama Desert, it protects the soil from sporadically occurring splash erosion and contributes to the accumulation of soil carbon and nitrogen as well as soil formation through bio‐weathering. The structure and function of the new CGC type are discussed, and we suggest the name grit–crust. We conclude that this type of CGC can be expected in all non‐polar fog deserts of the world and may resemble the cryptogam communities that shaped ancient Earth. It may thus represent a relevant player in current and ancient biogeochemical cycling.