פרסומים

The objectives were to test whether (1) melatonin blocks inhibition of embryonic development caused by heat shock at the zygote stage, and (2) the frequency of a thermoprotective allele for HSPA1L is increased in blastocysts formed from heat-shocked zygotes as compared with blastocysts from control zygotes. It was hypothesized that melatonin prevents effects of heat shock on development by reducing accumulation of reactive oxygen species (ROS) and that embryos inheriting the thermoprotective allele of HSPA1L would be more likely to survive heat shock. Effects of 1 µM melatonin on ROS were determined in experiments 1 and 2. Zygotes were cultured at 38.5 or 40°C for 3 h in the presence of CellROX reagent (ThermoFisher Scientific, Waltham, MA). Culture was in a low [5% (vol/vol)] oxygen (experiment 1) or low or high [21% (vol/vol)] oxygen environment (experiment 2). Heat shock and high oxygen increased ROS; melatonin decreased ROS. Development was assessed in experiments 3 and 4. In experiment 3, zygotes were cultured in low oxygen ± 1 µM melatonin and exposed to 38.5 or 40°C for 12 h (experiment 1) beginning 8 h after fertilization. Melatonin did not protect the embryo from heat shock. Experiment 4 was performed similarly except that temperature treatments (38.5 or 40°C, 24 h) were performed in a low or high oxygen environment (2×2 × 2 factorial design with temperature, melatonin, and oxygen concentration as main effects), and blastocysts were genotyped for a deletion (D) mutation (C→D) in the promoter region of HSPA1L associated with thermotolerance. Heat shock decreased percent of zygotes developing to the blastocyst stage independent of melatonin or oxygen concentration. Frequency of genotypes for HSPA1L was affected by oxygen concentration and temperature, with an increase in the D allele for blastocysts that developed in high oxygen and following heat shock. It was concluded that (1) lack of effect of melatonin or oxygen concentration on embryonic development means that the negative effects of heat shock on the zygote are not mediated by ROS, (2) previously reported effect of melatonin on fertility of heat-stressed cows might involve actions independent of the antioxidant properties of melatonin, and (3) the deletion mutation in the promoter of HSPA1L confers protection to the zygote from heat shock and high oxygen. Perhaps, embryonic survival during heat stress could be improved by selecting for thermotolerant genotypes.

The cosmopolitan coccolithophore Emiliania huxleyi is a unicellular eukaryotic alga responsible for vast blooms in the ocean. These blooms have immense impact on large biogeochemical cycles and are terminated by a specific large double-stranded DNA E. huxleyi virus (EhV, Phycodnaviridae). EhV infection is accompanied by induction of hallmarks of programmed cell death and production of reactive oxygen species (ROS). Here we characterized alterations in ROS metabolism and explored its role during infection. Transcriptomic analysis of ROS-related genes predicted an increase in glutathione (GSH) and H2O2 production during infection. In accordance, using biochemical assays and specific fluorescent probes we demonstrated the overproduction of GSH during lytic infection. We also showed that H2O2 production, rather than superoxide, is the predominant ROS during the onset of the lytic phase of infection. Using flow cytometry, confocal microscopy and multispectral imaging flow cytometry, we showed that the profound co-production of H2O2 and GSH occurred in the same subpopulation of cells but at different subcellular localization. Positively stained cells for GSH and H2O2 were highly infected compared with negatively stained cells. Inhibition of ROS production by application of a peroxidase inhibitor or an H2O2 scavenger inhibited host cell death and reduced viral production. We conclude that viral infection induced remodeling of the host antioxidant network that is essential for a successful viral replication cycle. This study provides insight into viral replication strategy and suggests the use of specific cellular markers to identify and quantify the extent of active viral infection during E. huxleyi blooms in the ocean.

In this report, we summarize and describe the recent unique updates and additions to the Molcas quantum chemistry program suite as contained in release version 8. These updates include natural and spin orbitals for studies of magnetic properties, local and linear scaling methods for the Douglas–Kroll–Hess transformation, the generalized active space concept in MCSCF methods, a combination of multiconfigurational wave functions with density functional theory in the MC-PDFT method, additional methods for computation of magnetic properties, methods for diabatization, analytical gradients of state average complete active space SCF in association with density fitting, methods for constrained fragment optimization, large-scale parallel multireference configuration interaction including analytic gradients via the interface to the Columbus package, and approximations of the CASPT2 method to be used for computations of large systems. In addition, the report includes the description of a computational machinery for nonlinear optical spectroscopy through an interface to the QM/MM package Cobramm. Further, a module to run molecular dynamics simulations is added, two surface hopping algorithms are included to enable nonadiabatic calculations, and the DQ method for diabatization is added. Finally, we report on the subject of improvements with respects to alternative file options and parallelization.

The STIL protein is essential for centriole replication and for the non-templated, de novo centriole biogenesis that is required for mammalian embryogenesis. Here we performed quantitative biophysical and structural analysis of the central short coiled coil domain (CCD) of STIL that is critical for its function. Using biophysical, biochemical and cell biology approaches, we identified the specific residues in the CCD that mediate the oligomerization, centrosomal localization and protein interactions of STIL. We characterized the structural properties of the coiled coil peptide using circular dichroism spectroscopy and size exclusion chromatography. We identified two regions in this domain, containing eight hydrophobic residues, which mediate the coiled coil oligomerization. Mutations in these residues destabilized the coiled coil thermodynamically but in most cases did not affect its secondary structure. Reconstituting mouse embryonic fibroblasts lacking endogenous Stil, we show that STIL oligomerization mediated by these residues is not only important for the centrosomal functions of STIL during the canonical duplication process but also for de-novo formation of centrosomes.

Molecularly imprinted polymer (MIP) particles offer many advantages as recognition objects. Therefore, we examd. thoroughly the effect of various exptl. parameters including cross-linker concn., nature of solvent, type of template and its concn., reaction vol. and temp. on the formation of particulate MIPs using pptn. polymn. The particles were characterized by various methods, such as XHR-SEM, HR-TEM and BET. Statistical calcns. were carried out using ImageJ software over population of at least 200 particles in each expt. The reproducibility of MIP synthesis was examd. and the SD was better than 10%. MIP particles were imprinted with different Sudan dye derivs. (Sudan II, III, IV and Sudan orange G). Raman and FTIR spectroscopies were used to prove the successful imprinting. Reuptake measurements were carefully compared between non-imprinted molecularly polymer (NIP) and MIP particles employing the same quantity and similar diam. and polydispersity of NIP and MIP, as never reported before. The results showed that MIP particles imprinted by Sudan IV showed higher selectivity toward this specific dye as compared with MIPs imprinted by other Sudan dyes and NIPs. [on SciFinder(R)]

Molecularly imprinted polymer (MIP) particles offer many advantages as recognition objects. Therefore, we examd. thoroughly the effect of various exptl. parameters including cross-linker concn., nature of solvent, type of template and its concn., reaction vol. and temp. on the formation of particulate MIPs using pptn. polymn. The particles were characterized by various methods, such as XHR-SEM, HR-TEM and BET. Statistical calcns. were carried out using ImageJ software over population of at least 200 particles in each expt. The reproducibility of MIP synthesis was examd. and the SD was better than 10%. MIP particles were imprinted with different Sudan dye derivs. (Sudan II, III, IV and Sudan orange G). Raman and FTIR spectroscopies were used to prove the successful imprinting. Reuptake measurements were carefully compared between non-imprinted molecularly polymer (NIP) and MIP particles employing the same quantity and similar diam. and polydispersity of NIP and MIP, as never reported before. The results showed that MIP particles imprinted by Sudan IV showed higher selectivity toward this specific dye as compared with MIPs imprinted by other Sudan dyes and NIPs. [on SciFinder(R)]

Mobility is one of the principle topics of humans, be it as nomads in former days, be it as frequent travellers for business or in leisure nowadays. Early in history, voyagers wrote their experiences down, but only very late travel stories become a respected desk in journalistic media. As special-interest-journalism it is and was highly coined by economic influence. The new book "Motor/Reise" - travel- and motor journalism - is part of the series "Journalismus Bibliothek", released by the renowned publisher Herbert von Halem Verlag, Cologne, Germany. The authors picture the history of travel- and mobility journalism from the ancient world until today. They consider new developments due to digital journalism - for instance travel blogs and mobile reporting -, ethical challenges, and the special role of photography in travel- and motorjournalism. Examples of best practice show freshmen and old-stagers how to engage in modern travel- and mobility journalism, and how to deal self-confidently with demands of makers, providers, and advertisers. Evelyn Runge and Hektor Haarkötter have intensivley worked as journalists themselves and clearly enrich their writing from these experiences.

(Herbert von Halem Verlag, Cologne ,Germany, 2016)

http://www.halem-verlag.de/motor-reise/

The terms Lower Palaeolithic and Middle Palaeolithic represent research constructs within which cultural evolution and prehistoric hominin behaviours can be studied, with the transition usually understood as marking a watershed in our evolution: an adaptation with a million-year record of success that gives way to something new. The interpretation of the Lower Palaeolithic Acheulian technocomplex is usually understood as a period of cultural stasis that extends over much of Africa and Eurasia, principally associated with Homo erectus. Those innovations that can be observed occur widely separated from one another in space and time. Yet a closer and more detailed examination of the Middle Pleistocene records from East Africa, southern Africa, Europe and the Levant reveals significant variation in cultural repertoires. A kind of paradox emerges, in which an Old World Lower Palaeolithic, apparently lacking an overall dynamic of distinctive and directed change in terms of cumulative variation over time, nevertheless culminates in a transition which sees the universal appearance of the Middle Palaeolithic. The two main hypotheses that have been advanced to explain the global transition, which happens essentially synchronously, appear mutually exclusive and contradictory. One view is that altered climatic-environmental constraints enabled and encouraged an ‘Out-of-Africa' dispersal (or dispersals) of a new type of genus Homo. This cultural replacement model has been challenged more recently by the alternative hypothesis of accumulating but unrelated and temporally non-linked regional, and in fact potentially autochthonous, processes. The Levant, by virtue of its position bridging Africa and Eurasia (thus being the region into which any out-of-Africa groups would have had first to disperse into), must be seen as a critical region for assessing the relative merits of these competing hypotheses. This paper deals with the Lower–Middle Paleolithic boundary in the Levant within a long temporal perspective. The Middle Pleistocene record in the Levant enables us to examine the amplitude of variation within each techno-complex, as well as to question whether there are diachronic changes in the amplitude of techno-typological variations as well as changes in the manner by which they appear in the record. The results carry significant implications for understandings of demographic and societal processes during the Lower–Middle Paleolithic transition in the Levant.

In today's modern pharmacologic approach to treating sight-threatening retinal vascular disorders, there is an increasing demand for a compact, mobile, lightweight and cost-effective fluorescein fundus camera to document the effects of antiangiogenic drugs on laser-induced choroidal neovascularization (CNV) in mice and other experimental animals. We have adapted the use of the Kowa Genesis Df Camera to perform Fundus Fluorescein Angiography (FFA) in mice. The 1 kg, 28 cm high camera has built-in barrier and exciter filters to allow digital FFA recording to a Compact Flash memory card. Furthermore, this handheld unit has a steady Indirect Lens Holder that firmly attaches to the main unit, that securely holds a 90 diopter lens in position, in order to facilitate appropriate focus and stability, for photographing the delicate central murine fundus. This easily portable fundus fluorescein camera can effectively record exceptional central retinal vascular detail in murine laser-induced CNV, while readily allowing the investigator to adjust the camera's position according to the variable head and eye movements that can randomly occur while the mouse is optimally anesthetized. This movable image recording device, with efficiencies of space, time, cost, energy and personnel, has enabled us to accurately document the alterations in the central choroidal and retinal vasculature following induction of CNV, implemented by argon-green laser photocoagulation and disruption of Bruch's Membrane, in the experimental murine model of exudative macular degeneration.