Genome-wide CRISPR Screens Reveal Host Factors Critical for SARS-CoV-2 Infection

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From Regional to National Clouds: TV Coverage in the Czech Republic

Media, and particularly TV media, have a great impact on the general public. In recent years, spatial patterns of information and the relevance of intangible geographies have become increasingly important. Gatekeeping plays a critical role in the selection of information that is transformed into media. Therefore, gatekeeping, through national media, also co-forms the generation of mental maps. In this paper, correspondence analysis (a statistical method) combined with cloud lines (a new visual analytics technique) is used to analyze how individual major regional events in one of the post-communist countries, the Czech Republic, penetrate into the media on a national scale. Although national news should minimize distortions about regions, this assumption has not been verified by our research. Impressions presented by the media of selected regions that were markedly influenced by one or several events in those regions demonstrate that gatekeepers, especially news reporters, functioned as a filter by selecting only a few specific, and in many cases, unusual events for dissemination.     

Mycobacterium tuberculosis ClpX Interacts with FtsZ and Interferes with FtsZ Assembly

FtsZ assembly at the midcell division site in the form of a Z-ring is crucial for initiation of the cell division process in eubacteria. It is largely unknown how this process is regulated in the human pathogen Mycobacterium tuberculosis. Here we show that the expression of clpX was upregulated upon macrophage infection and exposure to cephalexin antibiotic, the conditions where FtsZ-ring assembly is delayed. Independently, we show using pull-down, solid-phase binding, bacterial two-hybrid and mycobacterial protein fragment complementation assays, that M. tuberculosis FtsZ interacts with ClpX, the substrate recognition domain of the ClpXP protease. Incubation of FtsZ with ClpX increased the critical concentration of GTP-dependent polymerization of FtsZ. Immunoblotting revealed that the intracellular ratio of ClpX to FtsZ in wild type M. tuberculosis is approximately 1∶2. Overproduction of ClpX increased cell length and modulated the localization of FtsZ at midcell sites; however, intracellular FtsZ levels were unaffected. A ClpX-CFP fusion protein localized to the cell poles and midcell sites and colocalized with the FtsZ-YFP protein. ClpX also interacted with FtsZ mutant proteins defective for binding to and hydrolyzing GTP and possibly for interactions with other proteins. Taken together, our results suggest that M. tuberculosis ClpX interacts stoichiometrically with FtsZ protomers, independent of its nucleotide-bound state and negatively regulates FtsZ activities, hence cell division.     

Use of tissue culture to bypass wheat hybrid necrosis

Summary Hybrid necrosis in wheat is a barrier to gene transfer in wheat breeding practice. It is based on two complementary genes, Ne1 and Ne2. Recovery mutants (Re1, Re2 and Re3) which can grow to maturity were recovered from immature embryo cultures of necrotic hybrids between T. aestivum and T. durum. Cytological observation demonstrated that Re1 had 34 chromosomes instead of 35. This indicated that one of the chromosomes carrying the Ne genes was lost. Genetic study suggested that for Re1, the lost chromosome was chromosome 5B of the durum parental line. Re mutants are male sterile but can be maintained through a young ear culture method. Re mutants could be successfully pollinated by either parental line and the BC1 progeny is partially fertile. Re mutants were repeatedly induced in about 1% of the regenerated plants from immature embryo culture. This technique provides a practical way to bypass hybrid necrosis.     

Effects of GnRH on superovulated cattle

Gonadotropin releasing hormone (GnRH) was given to 109 cows and heifers during the course of 224 superovulations. Follicle stimulating hormone (FSH) was administered twice daily (5 or 6 mg) for 3.5 to 4 days beginning on any of Days 9 to 14 of the estrous cycle; prostaglandin (45 mg PGF(2)alpha or 750 ug cloprostenol) was given in a split dose on the fourth day. Donor cows and heifers were placed into four groups according to previous superovulation treatments, which consisted of one to three treatments or of no previous treatment. Every other cow or heifer within each of the four subgroups was treated with GnRH (200 mug i.m.) at standing estrus. Only donors that exhibited estrus within 32 to 72 h after the first prostaglandin treatment were used in the study. Animals were inseminated artificially 12 and 24 h after standing estrus was first observed. No differences were noted in the number of ovulations, total ova or transferable embryos recovered from the GnRH or control groups; however, two interactions were detected. Cows given GnRH had fewer palpable corpora lutea than control cows (P < 0.05), but this difference was not seen in heifers. The second interaction was that GnRH seemed to depress ovulation rate in donors not previously superovulated, but this effect was not observed with subsequent superovulations. Cows yielded more total ova than heifers (P < 0.01). There was no difference in return to estrus between GnRH and control groups after a second prostaglandin treatment at the time of embryo recovery. Most donors within each group resumed cycling between 5 and 12 d after embryo recovery.     

The critical region on the human Xq

Summary Adult female carriers of balanced X; autosome translocations (118 cases) and of balanced X inversions (31 cases) have been collected from the literature. Forty-five of the 118 translocation carriers in whom the break was in the critical region (Xq13–q22, Xq22–q26, separated by a narrow region within Xq22) showed gonadal dysgenesis. Seven of the 31 inversion carriers in whom the break was in the same region also had gonadal dysgenesis, whereas the remaining 24 were normal in this respect. The critical region consists mainly of Q-bright material, and is the fifth brightest segment in the human genome. The region contains relatively few genes. It is possible that meiotic crossing-over, rarely, if ever, takes place in it. The critical region may therefore consist of two supergenes whose integrity must be maintained to allow normal ovarian development. The effect exerted by this region differs from other known position effects, in that it is independent of the break-point within the region and of the chromosome bands to which the broken ends are attached. One possible mechanism causing this effect might be a change in the replication order of the chromosome bands, which, in turn, might affect their function.     

Behavior of sperm nuclei in intact and bisected metaphase II mouse oocytes fertilized in the presence of colcemid

Our objective was to examine the developmental fate of sperm nuclei in oocytes fertilized under conditions of meiotic arrest. Therefore zona-free metaphase II oocytes and oocyte fragments (nucleate and anucleate) were fertilized in the presence of colcemid. In anucleate oocyte fragments, normal male pronuclei develop. In contrast, in intact oocytes and nucleate fragments sperm nuclei after initial decondensation undergo secondary condensation. This state is maintained as long as the oocytes are treated with colcemid. When the drug is removed 3 h after insemination, the meiotic spindle(s) is reconstructed, the second polar body(ies) is extruded, and a female pronucleus (or micronuclei) forms. At the same time the sperm nucleus decondenses again and transforms into a male pronucleus. In addition oocytes fertilized in the presence of colcemid could not be refertilized. These observations suggest that oocytes and oocyte fragments fertilized in the presence of colcemid undergo activation despite the failure of pronucleus formation. The inhibitory effect of colcemid on the formation of pronuclei is expressed only in the presence of oocyte chromosomes. We suggest that colcemid stabilizes factors responsible for chromosome condensation that are associated with oocyte chromosomes but not factors (whether the same or different) present in the cytoplasm.     

Storage mites sensitivity and allergic respiratory disease

Summary In order to evaluate the frequency of skin sensitivity to Storage Mites and the role of such sensitization in respiratory allergic disease in workers with occupational exposure to stored items we studied 217 dock workers, 93 farmers and 104 white collars.From the results of skin prick tests the sensitization to sole Storage Mites appears significantly higher among people working in docks or farms, compared with a control group. This confirms the role of the working environment in inducing sensitization to Storage Mites. Rhinitis and asthma however affect nearly always (27/29 cases) people with an associated sensitization to House Dust Mites. Further studies are needed to define the allergenic importance of Storage Mites in working environments.