פרסומים

An unresolved question is how HIV-1 achieves efficient replication in terminally differentiated macrophages despite the restriction factor SAMHD1. We reveal inducible changes in expression of cell cycle-associated proteins including MCM2 and cyclins A, E, D1/D3 in macrophages, without evidence for DNA synthesis or mitosis. These changes are induced by activation of the Raf/MEK/ERK kinase cascade, culminating in upregulation of CDK1 with subsequent SAMHD1 T592 phosphorylation and deactivation of its antiviral activity. HIV infection is limited to these G1-like phase macrophages at the single-cell level. Depletion of SAMHD1 in macrophages decouples the association between infection and expression of cell cycle-associated proteins, with terminally differentiated macrophages becoming highly susceptible to HIV-1. We observe both embryo-derived and monocyte-derived tissue-resident macrophages in a G1-like phase at frequencies approaching 20%, suggesting how macrophages sustain HIV-1 replication in vivo Finally, we reveal a SAMHD1-dependent antiretroviral activity of histone deacetylase inhibitors acting via p53 activation. These data provide a basis for host-directed therapeutic approaches aimed at limiting HIV-1 burden in macrophages that may contribute to curative interventions.

Summary Emission of volatiles at advanced stages of flower development is a strategy used by plants to lure pollinators to the flower. We reveal that GA negatively regulates floral scent production in petunia. We used Agrobacterium-mediated transient expression of GA-20ox in petunia flowers and a virus-induced gene silencing approach to knock down DELLA expression, measured volatile emission, internal pool sizes and GA levels by GC-MS or LC–MS/MS, and analyzed transcript levels of scent-related phenylpropanoid-pathway genes. We show that GA has a negative effect on the concentrations of accumulated and emitted phenylpropanoid volatiles in petunia flowers; this effect is exerted through transcriptional/post-transcriptional downregulation of regulatory and biosynthetic scent-related genes. Both overexpression of GA20-ox, a GA-biosynthesis gene, and suppression of DELLA, a repressor of GA-signal transduction, corroborated GA's negative regulation of floral scent. We present a model in which GA-dependent timing of the sequential activation of different branches of the phenylpropanoid pathway during flower development may represent a link between the showy traits controlling pollinator attraction, namely color and scent.

A game of threats on a finite set of players, $N$, is a function $d$ that assigns a real number to any coalition, $S subseteq N$, such that $d left( S right) = - d left( N setminus S right)$. A game of threats is not necessarily a coalitional game as it may fail to satisfy the condition $d left( emptyset right) = 0$. We show that analogs of the classic Shapley axioms for coaltional games determine a unique value for games of threats. This value assigns to each player an average of the threat powers, $d left( S right)$, of the coalitions that include the player.

Mycobacterium tuberculosis is a major health threat, necessitating novel drug targets. Protein synthesis in bacteria uses initiator tRNAi charged with formylated methionine residue. Deletion of the formylase gene, tRNAfMet-formyl transferase (fmt), causes severe growth-retardation in E. coli and in S. pneumoniae, but not in P. aeruginosa or S. aureus. fmt was predicted to be essential in M. tuberculosis by transposon library analysis, but this was never formally tested in any mycobacteria. We performed a targeted deletion of fmt in M. smegmatis as well as Mtb-complex (M. bovis). In both cases, we created a mero-diploid strain, deleted the native gene by two-step allelic exchange or specialized-phage transduction, and then removed the complementing gene to create full deletion mutants. In M. smegmatis a full deletion strain could be easily created. In contrast, in M. bovis-BCG, a full deletion strain could only be created after incubation of 6 weeks, with a generation time 2 times longer than for wt bacteria. Our results confirm the importance of this gene in pathogenic mycobacteria, but as the deletion mutant is viable, validity of fmt as a drug target remains unclear. Our results also refute the previous reports that fmt is essential in M. tuberculosis-complex.

Complex ecosystems harbor multiple predators and prey species whose direct and indirect interactions are under study. In particular, the combined effects of predator diversity and resource preference on prey removal are not known. To understand the effect of interspecies interactions, combinations of micro-predators—i.e., protists (generalists), predatory bacteria (semi-specialists), and phages (specialists)—and bacterial prey were tracked over a 72-h period in miniature membrane bioreactors. While specialist predators alone drove their preferred prey to extinction, the inclusion of a generalist resulted in uniform losses among prey species. Most importantly, presence of a generalist predator enabled coexistence of all predators and prey. As the generalist predator also negatively affected the other predators, we suggest that resource partitioning between predators and the constant availability of resources for bacterial growth due to protist predation stabilizes the system and keeps its diversity high. The appearance of resistant prey strains and subsequent evolution of specialist predators unable to infect the ancestral prey implies that multitrophic communities are able to persist and stabilize themselves. Interestingly, the appearance of BALOs and phages unable to infect their prey was only observed for the BALO or phage in the absence of additional predators or prey species indicating that competition between predators might influence coevolutionary dynamics.

Replication of the genome occurs during S phase of the cell cycle in a highly regulated process that ensures the fidelity of DNA duplication. Each genomic region is replicated at a distinct time during S phase through the simultaneous activation of multiple origins of replication. Time of replication (ToR) correlates with many genomic and epigenetic features and is linked to mutation rates and cancer. Comprehending the full genomic view of the replication program, in health and disease is a major future goal and challenge. This article describes in detail the "Copy Number Ratio of S/G1 for mapping genomic Time of Replication" method (herein called: CNR-ToR), a simple approach to map the genome wide ToR of mammalian cells. The method is based on the copy number differences between S phase cells and G1 phase cells. The CNR-ToR method is performed in 6 steps: 1. Preparation of cells and staining with propidium iodide (PI); 2. Sorting G1 and S phase cells using fluorescence-activated cell sorting (FACS); 3. DNA purification; 4. Sonication; 5. Library preparation and sequencing; and 6. Bioinformatic analysis. The CNR-ToR method is a fast and easy approach that results in detailed replication maps.

Accelerated by the introduction of Next-Generation Sequencing (NGS), a number of genomes of cyprinid fish species have been drafted, leading to a highly valuable collective resource of comparative genome information on cyprinids (Cyprinidae). In addition, NGS-based transcriptome analyses of different developmental stages, organs, or cell types, increasingly contribute to the understanding of complex physiological processes, including immune responses. Cyprinids are a highly interesting family because they comprise one of the most-diversified families of teleosts and because of their variation in ploidy level, with diploid, triploid, tetraploid, hexaploid and sometimes even octoploid species. The wealth of data obtained from NGS technologies provides both challenges and opportunities for immunological research, which will be discussed here. Correct interpretation of ploidy effects on immune responses requires knowledge of the degree of functional divergence between duplicated genes, which can differ even between closely-related cyprinid fish species. We summarize NGS-based progress in analysing immune responses and discuss the importance of respecting the presence of (multiple) duplicated gene sequences when performing transcriptome analyses for detailed understanding of complex physiological processes. Progressively, advances in NGS technology are providing workable methods to further elucidate the implications of gene duplication events and functional divergence of duplicates genes and proteins involved in immune responses in cyprinids. We conclude with discussing how future applications of NGS technologies and analysis methods could enhance immunological research and understanding.

The geomorphic response of channels to base-level fall is an important factor in landscape evolution. To better understand the complex interactions between the factors controlling channel evolution in an emerging continental shelf setting, we use an extensive data set (high-resolution digital elevation models, aerial photographs, and Landsat imagery) of a newly incising, perennial segment of Nahal (Wadi) HaArava, Israel. This channel responds to the rapid and progressive lowering of its base-level, the Dead Sea ( \textgreater 30 m in \~35 years; \~0.5-1.3 m yr -1 ). Progressively evolving longitudinal profiles, channel width, sinuosity, and knickpoint retreat during the last few decades were documented or reconstructed. The results indicate that even under fast base-level fall, rapid delta progradation on top of the shelf and shelf edge can moderate channel mouth slopes and, therefore, largely inhibit channel incision and knickpoint propagation. This channel elongation stage ends when the delta reaches an extended accommodation within the receiving basin and fails to keep the channel mouth slopes as low as the channel bed slopes. Then, processes of incision, narrowing, and meandering begin to shape the channel and expand upstream. When the down-cutting channel encounters a more resistant stratum within the channel substrate, these processes are restricted to a downstream reach by formation of a retreating vertical knickpoint. When the knickpoint and the channel incise to a level below this stratum, a spatially continuous, diffusion-like evolution characterizes the channel’s response and source-to-sink transport can be implemented. These results emphasize the mouth slope and channel substrate resistance as the governing factors over long-term channel evolution, whereas flash floods have only local and short-lived impacts in a confined, continuously incising channel. The documented channel response applies to eustatic base-level fall under steepening basin bathymetry, rapid delta progradation, and lithologic variations in the channel substrate.

This study assessed the global hemostasis (including prothrombin time [PT], activated partial thromboplastin time [aPTT], antithrombin activity [ATA], fibrinogen and d-Dimer concentrations, platelet count, plateletcrit and thromboelastometry) in healthy pregnant bitches, comparing the results with those of healthy bitches at different estrous cycle stages, and assessed whether hemostatic changes during pregnancy are associated with serum progesterone concentration or the presence of fetuses in utero. The results show that pregnant bitches have higher fibrinogen concentration, platelet count and platelatecrit, and that fibrin and global clot formations occur faster than in non-pregnant bitches at different estrous cycle stages. Additionally, clot strength was higher in pregnant bitches than in non-pregnant ones. There were no differences in PT, ATA, and D-dimer concentration between all study groups. The aPTT was significantly shorter in bitches at the fourth and last pregnancy weeks, compared to the anestrus group, and shorter in both the fourth and last pregnancy weeks groups, compared to diestrus group. These results all support a hypercoagulable state in healthy pregnant bitches, unassociated with progesterone concentration.

There are numerous heritable diseases associated with mutations in the LMNA gene. Most of these laminopathic diseases, including several muscular dystrophies, are autosomal dominant and have tissue-specific phenotypes. Our previous studies have shown that the globally expressed Emery-Dreifuss muscular dystrophy (EDMD)-linked lamin mutation, L535P, disrupts nuclear mechanical response specifically in muscle nuclei of C. elegans leading to atrophy of the body muscle cells and to reduced motility. Here we used RNA sequencing to analyze the global changes in gene expression caused by the L535P EDMD lamin mutation in order to gain better understanding of disease mechanisms and the correlation between transcription and phenotype. Our results show changes in key genes and biological pathways that can help explain the muscle specific phenotypes. In addition, the differential gene expression between wild-type and L535P mutant animals suggests that the pharynx function in the L535P mutant animals is affected by this lamin mutation. Moreover, these transcriptional changes were then correlated with reduced pharynx activity and abnormal pharynx muscle structure. Understanding disease mechanisms will potentially lead to new therapeutic approaches toward curing EDMD.

The present study aims to describe existing peer-to-peer, social network-based sharing practices among adult students in teacher colleges.Ubiquitous social network sites open up a wide array of possibilities for peer-to-peer information and knowledge sharing. College instructors are often unaware of such practices that happen behind the scenes.An interpretative, qualitative research methodology was used. Thirty-seven Israeli students at a teacher college in Israel participated in either focus group discussions of (N = 29) or in-depth interviews (N = 8).Whereas knowledge sharing has been a main focus of research in organizational and information sciences, its relevance to educational settings has thus far been underscored. Recent research shows that peer–to-peer knowledge sharing is widespread among teenage students. The current study extends that work to an adult student population.The findings show thatknowledge sharing of this type is a common and even central feature of students’ college life and study behavior. It takes place through a variety of small and larger social network-based peer groups of different formations, including mostly college students but at time also practicing, experienced teachers. Sharing groups are formed on the spot for short term purposes or are stable, continuous over longer time periods. The contents shared are predominantly lesson summaries, material for exams, reading summaries and lesson plans. They are used immediately or stored for future use, as students have access to vast data bases of stored materials that have been compiled throughout the years by students of previous cohorts. Teacher students mentioned a range of reasons for sharing, and overall regard it very positive. However, some downsides were also acknowledged (i.e., superficial learning, exclusion, attentional overload and interruptions).College faculty and teaching staff should be cognizant and informed about these widespread peer-based knowledge sharing practices and consider whether perhaps changes in teaching formats and task assignments are required as a result.Future research should extend this work to other higher education settings, cultures and countries, and should map the perceptions of higher education teaching staff about peer-to-peer, online knowledge sharing.