A 36-year-old man presented to the hospital with a 2-day history of fever, sore throat, and fatigue 5 days after visiting Wuhan, China. His temperature on admission was 37.8°C (100.04°F). Pulmonary auscultation was normal. Laboratory studies showed a normal white blood cell count (4.6 × 10 9 /L) with a differential count of 53.1% neutrophils. The blood procalcitonin level was normal. Chest CT showed multiple peripheral ground-glass opacities in both lungs with more involvement of the left upper lobe, lingular segment (Figure a-c). At admission, the real-time fluorescence polymerase chain reaction (RT-PCR) assay of the sputum was negative for the 2019 novel coronavirus (2019-nCoV) nucleic acid.Repeat CT chest performed 3 days after admission showed transformation of ground-glass opacities to more consolidation (Figure d-f ). A repeat RT-PCR 2019-nCoV Images in a 36-year-old man with a 2-day history of fever, sore throat, and fatigue 5 days after visiting Wuhan, China, and a negative sputum real-time fluorescence polymerase chain reaction assay for the 2019 novel coronavirus. (a, b) Chest CT scans obtained at presentation show ground-glass opacities (red box) in the right upper lobe and the lingular segment and left lower lobe (b). (c) Volume rendering of chest CT scan obtained at admission. (d, e) CT scans obtained 3 days after admission show progression of ground-glass opacities to an atoll sign in the right upper lobe (red boxes in d) and left lower lobe consolidation (red boxes in e). (f) Volume rendering of chest CT scan obtained 3 days after admission shows the new areas of consolidation. See also Movies 1 and 2 (online)This copy is for personal use only. To order printed copies, contact reprints@rsna.org

dHaloferax mediterranei is able to accumulate the bioplastic poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) with more than 10 mol% 3-hydroxyvalerate (3HV) from unrelated carbon sources. However, the pathways that produce propionyl coenzyme A (propionyl-CoA), an important precursor of 3HV monomer, have not yet been determined. Bioinformatic analysis of H. mediterranei genome indicated that this strain uses multiple pathways for propionyl-CoA biosynthesis, including the citramalate/2-oxobutyrate pathway, the aspartate/2-oxobutyrate pathway, the methylmalonyl-CoA pathway, and a novel 3-hydroxypropionate pathway. Cofeeding of pathway intermediates and inactivating pathway-specific genes supported that these four pathways were indeed involved in the biosynthesis of 3HV monomer. The novel 3-hydroxypropionate pathway that couples CO 2 assimilation with PHBV biosynthesis was further confirmed by analysis of 13 C positional enrichment in 3HV. Notably, 13 C metabolic flux analysis showed that the citramalate/2-oxobutyrate pathway (53.0% flux) and the 3-hydroxypropionate pathway (30.6% flux) were the two main generators of propionyl-CoA from glucose. In addition, genetic perturbation on the transcriptome of the ⌬phaEC mutant (deficient in PHBV accumulation) revealed that a considerable number of genes in the four propionyl-CoA synthetic pathways were significantly downregulated. We determined for the first time four propionyl-CoA-supplying pathways for PHBV production in haloarchaea, particularly including a new 3-hydroxypropionate pathway. These results would provide novel strategies for the production of PHBV with controllable 3HV molar fraction. P olyhydroxyalkanoates (PHAs) are deposited as carbon and energy materials by many bacteria and haloarchaea under unbalanced growth conditions (1, 2). Due to their excellent biodegradability, biocompatibility, and mechanical properties, PHAs have received increased attention as excellent alternatives for petroleum-derived plastics (3). The physical and mechanical properties of PHAs are closely correlated to monomer composition. Among the various types of PHAs, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and poly-3-hydroxybutyrate (PHB) are the two most extensively studied ones. Due to the incorporation of 3-hydroxyvalerate (3HV) monomer, PHBV becomes more ductile and easier to process and thus possesses a wider range of industrial applications than PHB (4).PHBV is usually produced from acetyl coenzyme A (acetylCoA) and propionyl coenzyme A (propionyl-CoA) via a threestep process catalyzed by ␤-ketothiolase (PhaA/BktB), ␤-ketoacyl-CoA reductase (PhaB), and PHA synthase sequentially (5). Thus far, few bacteria, including Nocardia corallina (6), Rhodococcus spp. (7), a mutant of Ralstonia eutropha (8), several purple nonsulfur bacteria (9), and Bacillus circulans (10) were found to be able to produce propionyl-CoA for PHBV biosynthesis from single unrelated carbon sources. Most bacteria require the addition of propionate in the media to produce PHBV (11). Since propion...

The elongation growth of the mushroom stipe is a characteristic but not well-understood morphogenetic event of basidiomycetes. We found that extending native stipe cell walls of Coprinopsis cinerea were associated with the release of N-acetylglucosamine and chitinbiose and with chitinase activity. Two chitinases among all detected chitinases from C. cinerea, ChiE1 and ChiIII, reconstituted heatinactivated stipe wall extension and released N-acetylglucosamine and chitinbiose. Interestingly, both ChiE1 and ChiIII hydrolyze insoluble crystalline chitin powder, while other C. cinerea chitinases do not, suggesting that crystalline chitin components of the stipe cell wall are the target of action for ChiE1 and ChiIII. ChiE1-or ChiIII-reconstituted heat-inactivated stipe walls showed maximal extension activity at pH 4.5, consistent with the optimal pH for native stipe wall extension in vitro; ChiE1or ChiIII-reconstituted heat-inactivated stipe wall extension activities were associated with stipe elongation growth regions; and the combination of ChiE1 and ChiIII showed a synergism to reconstitute heat-inactivated stipe wall extension at a low action concentration. Field emission scanning electron microscopy (FESEM) images showed that the inner surface of acid-induced extended native stipe cell walls and ChiE1-or ChiIII-reconstituted extended heat-inactivated stipe cell walls exhibited a partially broken parallel microfibril architecture; however, these broken transversely arranged microfibrils were not observed in the unextended stipe cell walls that were induced by neutral pH buffer or heat inactivation. Double knockdown of ChiE1 and ChiIII resulted in the reduction of stipe elongation, mycelium growth, and heatsensitive cell wall extension of native stipes. These results indicate a chitinasehydrolyzing mechanism for stipe cell wall extension. IMPORTANCE A remarkable feature in the development of basidiomycete fruiting bodies is stipe elongation growth that results primarily from manifold cell elongation. Some scientists have suggested that stipe elongation is the result of enzymatic hydrolysis of cell wall polysaccharides, while other scientists have proposed the possibility that stipe elongation results from nonhydrolytic disruption of the hydrogen bonds between cell wall polysaccharides. Here, we show direct evidence for a chitinase-hydrolyzing mechanism of stipe cell wall elongation in the model mushroom Coprinopsis cinerea that is different from the expansin nonhydrolysis mechanism of plant cell wall extension. We presumed that in the growing stipe cell walls, parallel chitin microfibrils are tethered by ␤-1,6-branched ␤-1,3-glucans, and that the breaking of the tether by chitinases leads to separation of these microfibrils to increase their spacing for insertion of new synthesized chitin and ␤-1,3-glucans under turgor pressure in vivo.O ne of the remarkable characteristics of the development of basidiomycete fruiting bodies is stipe elongation growth that results primarily from manifold cell elongation (1-5). The ...

Highlights d A subset of high-grade glioma-associated microglia (HGG-AM) is identified by scRNA-seq d TGF-b1 activated from SETD2-mut/IDH-WT GBM cells promotes activation of HGG-AM d HGG-AM exhibits pro-inflammation and proliferation features, promoting tumor progression

Background Medulloblastoma (MB) with metastases at diagnosis and recurrence correlates with poor prognosis. Unfortunately, the molecular mechanism underlying metastases growth has received less attention than primary therapy-naïve MB. Though astrocytes have been frequently detected in brain tumors, their roles in regulating the stemness properties of MB stem-like cells (MBSCs) in disseminated lesions remain elusive. Methods Effects of tumor-associated astrocyte (TAA)–secreted chemokine C-C ligand 2 (CCL2) on MBSC self-renewal was determined by immunostaining analysis. Necroptosis of TAA was examined by measuring necrosome activity. Alterations in Notch signaling were examined after inhibition of CCL2. Progression of MBSC-derived tumors was evaluated after pharmaceutical blockage of necroptosis. Results TAA, as the essential components of disseminated tumor, produced high levels of CCL2 to shape the inflammation microenvironment, which stimulated the enrichment of MBSCs in disseminated MB. In particular, CCL2 played a pivotal role in maintaining stem-like properties via Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3)–mediated activation of Notch signaling. Loss of CCL2/C-C chemokine receptor 2 (CCR2) function repressed the JAK2/STAT3-Notch pathway and impaired MBSC proliferation, leading to a dramatic reduction of stemness, tumorigenicity, and metastasizing capability. Furthermore, necroptosis-induced CCL2 release depended on activation of receptor-interacting protein 1 (RIP1)/RIP3/mixed lineage kinase domain-like pseudokinase (MLKL) in TAA, which promoted the oncogenic phenotype. Blockade of necroptosis resulted in CCL2 deprivation and compromised MBSC self-proliferation, indicating MBSCs outsourced CCL2 from necroptotic TAA. Finally, CCL2 was upregulated in high-risk stages of MB, further supporting its value as a prognostic indicator. Conclusion These findings highlighted the critical role of CCL2/CCR2 in Notch signaling activation in MBSCs and revealed a necroptosis-associated glial cytokine microenvironment driving stemness maintenance in disseminations. Key Points 1. TAA-derived CCL2 promoted stemness in disseminated MBSCs through Notch signaling activation via the JAK2/STAT3 pathway. 2. TAA released CCL2 in a RIP1/RIP3/MLKL-dependent manner leading to necroptosis.

Bacterial wilt caused by Ralstonia solanacearum is the most serious soilborne disease of tobacco (Nicotiana tabacum) in China. In this study, 89 strains were collected in 2012 to 2014 from across the four major tobacco-growing areas in China. The strains were identified as phylotype I by multiplex polymerase chain reaction and further divided into seven sequevars based on polymorphisms in the endoglucanase (egl) gene. Among the seven sequevars, four (15, 17, 34, and 44) have been previously described as pathogens of tobacco and two (13 and 14), which are reported here on tobacco, were previously found only on other plants. In addition, a new sequevar named 54 was identified. Strains from tobacco from different regions showed different levels of genetic diversity based on partial egl gene sequences. The farther north the distribution, the lower the gene diversity found. Pathogenicity of 27 representative strains was assessed by inoculation onto three tobacco cultivars of varying susceptibility. Through cluster analysis of area under the disease progress curve values, the 27 strains were classified into different pathotypes based on virulence; however, no obvious associations were found between sequevar and pathotype. These results will assist in determining geographical distribution of strains, and provide the foundation for breeding and integrated management programs in China.

BackgroundNeuroendocrine tumor originates from the diffuse neuroendocrine system. Intracranial originating is lower to 0.74 %.Case presentationWe present two cases of primary intracranial neuroendocrine tumor A 39-year-old woman was admitted with headache, fever, polydipsia and polyuria. Biochemical and endocrinological results showed hyponatremia, hypothyroidism and hypopituitarism. MRI scans demonstrated an obviouslyenhancing lesion in seller and superseller area. Then a gross removal of tumor was achieved during the single nostril transsphenoidal approach surgery. Pathological diagnosis was high-grade small-cell neuroendocrine tumor. A 40-year-old woman presented with multiple symptoms and neurological deficit. Neuroimaging results demonstrated a huge obviously-enhancing tumor in anterior cranial fossa. Biochemical and hormone findings revealed hypokalemia, high glucose and hypercortisolemia. The intracranial surgery achieved a gross removal through a right frontal craniotomy. Pathological diagnosis was low-grade small-cell neuroendocrine tumor with immuno-negativity for ACTH.ConclusionThe mechanism, diagnosis, and treatment of neuroendocrine tumor are still challenging.

Medulloblastoma is the most common malignant brain tumor in children. Although accumulated research has suggested that cancer stem-like cells play a key role in medulloblastoma tumorigenesis, the specific molecular mechanism regarding proliferation remains elusive. Here, we reported more abundant expression of maternal embryonic leucine-zipper kinase (MELK) and enhancer of zeste homolog 2 (EZH2) in medulloblastoma stemlike cells than in neural stem cells and the interaction between the two proteins could mediate the self-renewal of sonic hedgehog subtype medulloblastoma. In human medulloblastoma, extensive nodularity and large-cell/anaplastic subgroups differed according to the staining levels of MELK and EZH2 from the other two subgroups. The proportion of MELK-or EZH2-positive staining status could be considered as a potential indicator for survival. Mechanistically, MELK bound to and phosphorylated EZH2, and its methylation was induced by EZH2 in medulloblastoma, which could regulate the proliferation of cancer stemlike cells. In xenografts, loss of MELK or EZH2 attenuated medulloblastoma stem-like cell-derived tumor growth and promoted differentiation. These findings indicate that MELK-induced phosphorylation and EZH2-mediated methylation in MELK/EZH2 pathway are essential for medulloblastoma stem-like cell-derived tumor proliferation, thereby identifying a potential therapeutic strategy for these patients.Implications: This study demonstrates that the interaction occurring between MELK and EZH2 promotes self-proliferation and stemness, thus representing an attractive therapeutic target and potential candidate for diagnosis of medulloblastoma.Mol Cancer Res; 15(9); 1275-86. Ó2017 AACR.