Identification of male sterility-related genes in Saccharum officinarum and Saccharum spontaneum.

KEY MESSAGE: Candidate male sterility genes were identified in sugarcane, which interacts with kinase-related proteins, transcription factors, and plant hormone signaling pathways to regulate stamen and anther development. Saccharum officinarum is a cultivated sugarcane species that its predominant feature is high sucrose content in stems. Flowering is necessary for breeding new cultivars but will terminate plant growth and reduce sugar yield. The wild sugarcane species Saccharum spontaneum has robust and viable pollen, whereas most S. officinarum accessions are male sterile, which is a desirable trait of a maternal parent in sugarcane breeding. To study male sterility and related regulatory pathways in sugarcane, we carried out RNAseq using flowers in different developmental stages between male-sterile S. officinarum accession 'LA Purple' and fertile S. spontaneum accession 'SES208'. Gene expression profiles were used to detect how genes are differentially expressed between male sterile and fertile flowers and to identify candidate genes for male sterility. Weighted gene correlation networks analysis (WGCNA) was conducted to investigate the regulatory networks. Transcriptomic analyses showed that 988 genes and 2888 alleles were differentially expressed in S. officinarum compared to S. spontaneum. Ten differentially expressed genes and thirty alleles were identified as candidate genes and alleles for male sterility in sugarcane. The gene Sspon.03G0007630 and two alleles of the gene Sspon.08G0002270, Sspon.08G0002270-2B and Sspon.08G0014700-1A, were involved in the early stamen or carpel development stages, while the remaining genes were classified into the post-meiosis stage. Gibberellin, auxin, and jasmonic acid signaling pathways are involved in the stamen development in sugarcane. The results expanded our knowledge of male sterility-related genes in sugarcane and generated genomic resources to facilitate the selection of ideal maternal parents to improve breeding efficiency.

Expanded field of view frequency-selective incoherent holography by using a triple-beam setup.

We propose a new, to the best of our knowledge, method of incoherent optical frequency selection called three-pack frequency-selective incoherent holography. Compressed holography is reconstructed using phase shift intercepts and spatial transfer function convolution in the form of separation without loss of magnification or resolution. The frequency-selective reconstruction process removes the conjugate and DC terms along with the interception of the object wave. This work attempts three-dimensional reconstruction and selected-frequency phase extraction of axial slices in submicron steps, and the experimental results show the potential of the proposed method in areas such as compressed holography, extended field of view, and slice tomography.

Gastrodin ameliorates the lipopolysaccharide-induced neuroinflammation in mice by downregulating miR-107-3p.

Background: Neuroinflammation plays a pivotal role in the pathogenesis of Central Nervous System (CNS) diseases. The phenolic glucoside gastrodin (GAS), has been known to treat CNS disorders by exerting anti-inflammatory activities. Our aim was to investigate the potential neuroprotective mechanisms of GAS on lipopolysaccharide (LPS)-induced mice. Methods: Male C57BL/6J mice were treated by LPS, before which GAS was adminisrated. The behavior tests such as forced swim test, tail suspension test, and elevated plus maze were performed to evaluate depressive-anxiety-like behaviors. A high-throughput sequencing (HTS) analysis was performed to screen out distinctive miRNAs which were validated using quantitative real-time PCR. Then, miRNA agomir or NC was injected stereotaxically into hippocampus of mice to explore the role of miRNA on GAS in response to LPS. Furthermore, Immunofluorescence and the hematoxylin and eosin (H&E) staining were employed to observe the cellular morphology. The protein levels of pro-inflammatory factors were evaluated by western blot. Finally, the target mRNA of miRNA was predicted using bioinformatics analysis. GO and KEGG enrichment analyses were conducted to clarify the potential function of target protein, which were visualized by bubble charts. Results: The behavioral data showed that mice in the LPS group had obvious depressive-anxiety-like behaviors, and 100 mg/kg GAS could improve these behavioral changes and alleviate the levels of pro-inflammatory cytokines in the hippocampus when mice were exposed to LPS for 6 h. Meanwhile, LPS-induced microglia and astrocyte activation in the CA1, CA2, CA3, and DG regions of the hippocampus were also reversed by GAS. Furthermore, miR-107-3p were screened out and verified for GAS in response to LPS. Importantly, miR-107-3p overexpression negatively abrogated the neuroprotective effects of GAS. Moreover, KPNA1 might be the target molecular of miR-107-3p. KPNA1 might regulate 12 neuroinflammation-related genes, which were mainly involved in cytokine-mediated signaling pathway. Conclusion: These results suggested that GAS might alleviate the LPS-induced neuroinflammation and depressive-anxiety-like behaviors in mice by downregulating miR-107-3p and upregulating the downstream target KPNA1. The indicates miR-107-3p may provide a new strategy for the treatment of CNS diseases.

Auxin and cytokinin mediated regulation involved in vitro organogenesis of papaya.

In vitro organogenesis is a multistep process which is largely controlled by the balance between auxin and cytokinin. Previous studies revealed a complex network regulating in vitro organogenesis in Arabidopsis thaliana; however, our knowledge of the molecular mechanisms underlying de novo shoot formation in papaya (Carica papaya) remains limited. Here, we optimized multiple factors to achieve an efficient and reproducible protocol for the induction of papaya callus formation and shoot regeneration. Subsequently, we analyzed the dynamic transcriptome profiles of samples undergoing this process, identified 5381, 642, 4047, and 2386 differentially expressed genes (DEGs), including 447, 66, 350, and 263 encoding transcription factors (TFs), in four stage comparisons. The DEGs were mainly involved in phytohormone modulation and transduction processes, particularly for auxin and cytokinin. Of these, 21 and 7 candidate genes involved in the auxin and cytokinin pathways, respectively, had distinct expression patterns throughout in vitro organogenesis. Furthermore, we found two genes encoding key TFs, CpLBD19 and CpESR1, were sharply induced on callus induction medium and shoot induction medium, indicating these two TFs may serve as proxies for callus induction and shoot formation in papaya. We therefore report a regulatory network of auxin and cytokinin signaling in papaya according to the one previously modeled for Arabidopsis. Our comprehensive analyses provide insight into the early molecular regulation of callus initiation and shoot formation in papaya, and are useful for the further identification of the regulators governing in vitro organogenesis.

Ultra-long DNA molecule isolation from plant nuclei for ultra-long read genome sequencing.

Long and ultra-long read DNA sequencing technologies require high molecular weight DNA with high quality and sufficient quantity, which could be challenging to obtain from recalcitrant plant tissues. We describe a protocol to isolate ultra-long DNA from 12 species for ultra-long read genome sequencing. A suitable nuclei lysis buffer is critical for DNA quality and yield. This protocol will enable individual labs to isolate high molecular weight DNA at a rapid pace with low cost from a variety of plant species. For complete information on the use and execution of this protocol, please refer to: Zhang et al. (2020).

Low-frequency electroencephalogram oscillations govern left-eye lateralization during anti-predatory responses in the music frog.

Visual lateralization is widespread for prey and anti-predation in numerous taxa. However, it is still unknown how the brain governs this asymmetry. In this study, we conducted behavioral and electrophysiological experiments to evaluate anti-predatory behaviors and dynamic brain activities in Emei music frogs (Nidirana daunchina), to explore the potential eye bias for anti-predation and the underlying neural mechanisms. To do this, predator stimuli (a model snake head and a leaf as a control) were moved around the subjects in clockwise and anti-clockwise directions at steady velocity. We counted the number of anti-predatory responses and measured electroencephalogram (EEG) power spectra for each band and brain area (telencephalon, diencephalon and mesencephalon). Our results showed that (1) no significant eye preferences could be found for the control (leaf); however, the laterality index was significantly lower than zero when the predator stimulus was moved anti-clockwise, suggesting that left-eye advantage exists in this species for anti-predation; (2) compared with no stimulus in the visual field, the power spectra of delta and alpha bands were significantly greater when the predator stimulus was moved into the left visual field anti-clockwise; and, (3) generally, the power spectra of each band in the right-hemisphere for the left visual field were higher than those in the left counterpart. These results support that the left eye mediates the monitoring of a predator in music frogs and lower-frequency EEG oscillations govern this visual lateralization.

Positional cloning and characterization of the papaya diminutive mutant reveal a truncating mutation in the CpMMS19 gene.

The papaya diminutive mutant exhibits miniature stature, retarded growth and reduced fertility. This undesirable mutation appeared in the variety 'Sunset', the progenitor of the transgenic line 'SunUp', and was accidentally carried forward into breeding populations. The diminutive mutation was mapped to chromosome 2 and fine mapped to scaffold 25. Sequencing of a bacterial artificial chromosome in the fine mapped region led to the identification of the target gene responsible for the diminutive mutant, a gene orthologous to MMS19 with a 36.8 kb deletion co-segregating with the diminutive mutant. The genomic sequence of CpMMS19 is 62 kb, consisting of 20 exons and 19 introns. It encodes a protein of 1143 amino acids while the diminutive allele encodes a truncated protein of 287 amino acids. Expression of the full-length CpMMS19 was able to complement the thermosensitive growth of the yeast mms19 deletion mutant while expression of the diminutive allele resulted in increased thermosensitivity. Over-expression of the diminutive allele in Arabidopsis met18 mutant results in a high frequency of seed abortion. The papaya diminutive phenotype is caused by an alteration in gene function rather than a loss-of-function mutation. SCAR (sequence characterized amplified region) markers were developed for rapid detection of the diminutive allele in breeding populations.

Stratification of portal vein-invaded hepatocellular carcinoma treated with transarterial chemoembolization monotherapy.

PURPOSE: The study aimed to establish a prognostic prediction model and an artificial neural network (ANN) model to determine who will benefit from transarterial chemoembolization (TACE) monotherapy for patients with hepatocellular carcinoma (HCC) invading portal vein. METHODS: Treatment-naïve patients with HCC and portal vein invasion who were treated with TACE monotherapy at hospital A as training cohort and hospital B as validation cohort were included. The primary endpoint was overall survival (OS). In training cohort, independent risk factors associated with OS were identified by univariate and multivariate analysis. The prognostic prediction (PP) and ANN models based on the independent risk factors were established to find out who will benefit most from TACE monotherapy. The type of portal vein tumor thrombosis was classified based on the Cheng's Classification. The accuracy of the models was validated in validation cohort. RESULTS: Totally, 242 patients (training cohort: n â€‹= â€‹159; validation cohort: n â€‹= â€‹83) were included. The median OS was 7.1 and 8.5 months in training and validation cohort, respectively. In training cohort, the PP model was established based on the following five independent risk factors: Cheng's Classification, Eastern Cooperative Oncology Group score, maximum tumor size, number of HCC nodules, and Child-Pugh stage. PP score of 17.5 was identified as cut-off point and patients were divided into two groups by PP score <17.5 and >17.5 in survival benefit and prognostication (8.8 vs. 5.5 months; P â€‹< â€‹0.001). These five factors were included and ranked based on the importance associated with OS in the ANN model. Both of the two models received high accuracy after validation. CONCLUSIONS: Portal vein invaded HCC patients with PP score <17.5 may benefit most from TACE monotherapy. For these patients, TACE monotherapy should be considered.

Dynamics of electroencephalogram oscillations underlie right-eye preferences in predatory behavior of the music frog.

Visual lateralization is a typical characteristic of many vertebrates; however, its underlying dynamic neural mechanism is unclear. In this study, predatory responses and dynamic brain activities were evaluated in the Emei music frog (Nidirana daunchina) to assess the potential eye preferences and their underlying dynamic neural mechanism, using behavioral and electrophysiological experiments, respectively. To do this, when the prey stimulus (live cricket and leaf as control) was moved around the frogs in both clockwise and anticlockwise directions at constant velocity, the number of predatory responses were counted and electroencephalogram (EEG) absolute power spectra for each band were measured for the telencephalon, diencephalon and mesencephalon. The results showed that: (1) no significant differences in the number of predatory responses could be found for the control (leaf), but the number of predatory responses for the right visual field (RVF) was significantly greater than that for the left visual field (LVF) when the live cricket was moved into the RVF clockwise; (2) compared with no stimulus in the visual field and stimulus in the LVF, the power spectra of each EEG band were greater when the prey stimulus was moved into the RVF clockwise; and (3) the power spectra of the theta, alpha and beta bands in the left diencephalon were significantly greater than those of the right counterpart for the clockwise direction, but similar significant differences presented for the delta, theta and alpha bands in the anticlockwise direction. Together, the results suggested that right-eye preferences for predatory behaviors exist in music frogs, and that the dynamics of EEG oscillations might underlie this right eye/left hemisphere advantage.

Exploring the Effects of Online Rating and the Activeness of Physicians on the Number of Patients in an Online Health Community.

Introduction: With the rapid development of information technology and online communities, patients in growing numbers choose to consult physicians in online health communities (OHCs) for information and treatment. Although many researchers have examined OHCs, the impact of physicians' online rating and activeness on patient consultation choice, which is reflected by the number of patients in OHCs, has rarely been discussed.Methods: A computer program was developed to download the required data. We collected data from 8,401 physicians from an online health community, Good Doctor Online, and analyzed data with Stata. An empirical model was conducted to explore the factors that influence the number of patients who consult a physician in OHCs.Results: The results indicate that the online rating and activeness of physicians have positive effects on the number of patients. Furthermore, the physician's professional seniority and hospital rank significantly moderate this positive relationship. As hospital levels increase, the impacts of the online rating and activeness on the number of patients decrease. In addition, the online rating has a weaker influence on the number of patients when a physician has a high professional seniority.Conclusions: It is important to understand what factors influence the number of patients who choose to consult physicians in OHCs. This study aims to explore the effects of online rating and activeness of physicians on the number of patients, as well as the moderating effects of the physician's professional seniority and hospital rank. Our results have implications for existing health management and e-health literature, and OHC designers.

Comparative Analysis of Homologous Sequences of Saccharum officinarum and Saccharum spontaneum Reveals Independent Polyploidization Events.

Sugarcane (Saccharum spp. hybrids) is an economically important crop widely grown in tropical and subtropical regions for sugar and ethanol production. However, the large genome size, high ploidy level, interspecific hybridization and aneuploidy make sugarcane one of the most complex genomes and have long hampered genome research in sugarcane. Modern sugarcane cultivars are derived from interspecific hybridization between S. officinarum and S. spontaneum with 80-90% of the genome from S. officinarum and 10-20% of the genome from S. spontaneum. We constructed bacterial artificial chromosome (BAC) libraries of S. officinarum variety LA Purple (2n = 8x = 80) and S. spontaneum haploid clone AP85-441 (2n = 4x = 32), and selected and sequenced 97 BAC clones from the two Saccharum BAC libraries. A total of 5,847,280 bp sequence from S. officinarum and 5,011,570 bp from S. spontaneum were assembled and 749 gene models were annotated in these BACs. A relatively higher gene density and lower repeat content were observed in S. spontaneum BACs than in S. officinarum BACs. Comparative analysis of syntenic regions revealed a high degree of collinearity in genic regions between Saccharum and Sorghum bicolor and between S. officinarum and S. spontaneum. In the syntenic regions, S. spontaneum showed expansion relative to S. officinarum, and both S. officinarum and S. spontaneum showed expansion relative to sorghum. Among the 75 full-length LTR retrotransposons identified in the Saccharum BACs, none of them are older than 2.6 mys and no full-length LTR elements are shared between S. officinarum and S. spontaneum. In addition, divergence time estimated using a LTR junction marker and a syntenic gene shared by 3 S. officinarum and 1 S. spontaneum BACs revealed that the S. spontaneum intergenic region was distant to those from the 3 homologous regions in S. officinarum. Our results suggested that S. officinarum and S. spontaneum experienced at least two rounds of independent polyploidization in each lineage after their divergence from a common ancestor.

Radiofrequency ablation for hepatic hemangiomas: A consensus from a Chinese panel of experts.

Recent studies have shown that radiofrequency (RF) ablation therapy is a safe, feasible, and effective procedure for hepatic hemangiomas, even huge hepatic hemangiomas. RF ablation has the following advantages in the treatment of hepatic hemangiomas: minimal invasiveness, definite efficacy, high safety, fast recovery, relatively simple operation, and wide applicability. It is necessary to formulate a widely accepted consensus among the experts in China who have extensive expertise and experience in the treatment of hepatic hemangiomas using RF ablation, which is important to standardize the application of RF ablation for the management of hepatic hemangiomas, regarding the selection of patients with suitable indications to receive RF ablation treatment, the technical details of the techniques, therapeutic effect evaluations, management of complications, etc. A final consensus by a Chinese panel of experts who have the expertise of using RF ablation to treat hepatic hemangiomas was reached by means of literature review, comprehensive discussion, and draft approval.