A dominant negative 14-3-3 mutant in Schizosaccharomyces pombe distinguishes the binding proteins involved in sexual differentiation and check point.

The homothallic fission yeast Schizosaccharomyces pombe undergoes sexual differentiation when starved, but sam (skips the requirement of starvation for mating) mutants such as those carrying mutations in adenylate cyclase (cyr1) or protein kinase A (pka1) mate without starvation. Here, we identified sam3, a dominant negative allele of rad24, encoding one of two 14-3-3 proteins. Genetic mapping and whole-genome sequencing showed that the sam3 mutation comprises a change in nucleotide at position 959 from guanine to adenine, which switches the amino acid at position 185 from glutamic acid to lysine (E185K). We generated the rad24-E185K integrated mutant and its phenotype was similar to that of the sam3 mutant, including calcium sensitivity and UV non-sensitivity, but the phenotype is different from that of the Δrad24 strain. While the UV-sensitive phenotype was observed in the Δrad24 mutant, it was not observed in the sam3 and rad24-E185K mutants. The expression of the rad24-E185K gene in wild type cells induced spore formation in the nutrient rich medium, confirming rad24-E185K is dominant. This dominant effect of rad24-E185K was also observed in Δras1 and Δbyr2 diploid mutants, indicating that rad24-E185K generate stronger phenotype than rad24 null mutants. Ste11, the key transcription factor for sexual differentiation was expressed in sam3 mutants without starvation and it predominantly localized to the nucleus. The Rad24-E185K mutant protein retained its interaction with Check point kinase1 (Chk1), whereas it reduced interaction with Ste11, an RNA binding protein Mei2, and a MAPKKK Byr2, freeing these proteins from negative regulation by Rad24, that account for the sam phenotype and UV non-sensitive phenotype. Glucose depletion in rad24-E185K or Δpka1 Δrad24 double mutation induced haploid meiosis, leading to the formation of spores in haploid. The position of glutamic acid 185 is conserved in all major 14-3-3s; hence, our finding of a dominant negative allele of 14-3-3 is useful for understanding 14-3-3s in other organisms.

Automated unruptured cerebral aneurysms detection in TOF MR angiography images using dual-channel SE-3D UNet: a multi-center research.

OBJECTIVES: Time of flight magnetic resonance angiography (TOF-MRA) is the primary non-invasive screening method for cerebral aneurysms. We aimed to develop a computer-aided aneurysm detection method to improve the diagnostic efficiency and accuracy, especially decrease the false positive rate. METHODS: This is a retrospective multicenter study. The dataset contained 1160 TOF-MRA examinations composed of unruptured aneurysms (n = 1096) and normal controls (n = 166) from six hospitals. A total of 1037 examinations acquired from 2013 to 2019 were used as training set; 123 examinations acquired from 2020 to 2021 were used as external test set. We proposed an equalized augmentation strategy based on aneurysm location and constructed a detection model based on dual channel SE-3D UNet. The model was trained with a 5-fold cross-validation in the training set, then tested on the external test set. RESULTS: The proposed method achieved 82.46% sensitivity on patient-level, 73.85% sensitivity on lesion-level, and 0.88 false positives per case in the external test set. The performance did not show significant differences in subgroups according to the aneurysm site (except ACA), aneurysm size (except smaller than 3 mm), or MRI scanners. The performance preceded the basic SE-3D UNet by increasing 15.79% patient-level sensitivity and decreasing 4.19 FPs/case. CONCLUSIONS: The proposed automated aneurysm detection method achieved acceptable sensitivity while controlling fairly low false positives per case. It might provide a useful auxiliary tool of cerebral aneurysms MRA screening. KEY POINTS: • The need for automated cerebral aneurysms detecting is growing. • The strategy of equalized augmentation based on aneurysm location and dual-channel input could improve the model performance. • The retrospective multi-center study showed that the proposed automated cerebral aneurysms detection using dual-channel SE-3D UNet could achieve acceptable sensitivity while controlling a low false positive rate.

Neurotransmitter-mediated collective rhythms in grouped Drosophila circadian clocks.

Over the past decades, fly Drosophila melanogaster has being used as a premier model organism to study molecular and genetic bases of circadian rhythms. Here the authors propose a multicellular heterogeneous model for which the network of Drosophila circadian oscillators consists of two groups, the self-sustained lateral neurons (LNs) communicating to each other and the damped dorsal neurons (DNs) receiving neurotransmitters only from the LNs without interaction within this group. By simulating different experimental conditions, the authors find that the proposed model, except for being capable of reproducing some known experimental results well, also can predict some interesting phenomena: 1) The DNs need neuronal projections from the LNs to be rhythmic and to synchronize; 2) the effect of communication on mean amplitude and mean period of two oscillatory groups is different; 3) communication delay can facilitate the network synchronization of the LNs; and 4) only the LNs lose rhythmicity under constant light conditions. These results reveal the mechanism of an integrated pacemaker that would govern behavioral and physiological rhythmicity of the model organism.