Liquid chromatography-mass spectrometry-based metabolomics and fluxomics reveals the metabolic alterations in glioma U87MG multicellular tumor spheroids versus two-dimensional cell cultures.

RATIONALE: Multicellular tumor spheroids (MCTSs) that reconstitute the metabolic characteristics of in vivo tumor tissue may facilitate the discovery of molecular biomarkers and effective anticancer therapies. However, little is known about how cancer cells adapt their metabolic changes in complex three-dimensional (3D) microenvironments. Here, using the two-dimensional (2D) cell model as control, the metabolic phenotypes of glioma U87MG multicellular tumor spheroids were systematically investigated based on static metabolomics and dynamic fluxomics analysis. METHODS: A liquid chromatography-mass spectrometry-based global metabolomics and lipidomics approach was adopted to survey the cellular samples from 2D and 3D culture systems, revealing marked molecular differences between them. Then, by means of metabolomic pathway analysis, the metabolic pathways altered in glioma MCTSs were found using 13 C6 -glucose as a tracer to map the metabolic flux of glycolysis, the tricarboxylic acid (TCA) cycle, de novo nucleotide synthesis, and de novo lipid biosynthesis in the MCTS model. RESULTS: We found nine metabolic pathways as well as glycerolipid, glycerophospholipid and sphingolipid metabolism to be predominantly altered in glioma MCTSs. The reduced nucleotide metabolism, amino acid metabolism and glutathione metabolism indicated an overall lower cellular activity in MCTSs. Through dynamic fluxomics analysis in the MCTS model, we found that cells cultured in MCTSs exhibited increased glycolysis activity and de novo lipid biosynthesis activity, and decreased the TCA cycle and de novo purine nucleotide biosynthesis activity. CONCLUSIONS: Our study highlights specific, altered biochemical pathways in MCTSs, emphasizing dysregulation of energy metabolism and lipid metabolism, and offering novel insight into metabolic events in glioma MCTSs.

Aberrant METTL14 gene expression contributes to malignant transformation of benzene-exposed myeloid cells.

Benzene is a known contributor to human leukaemia through its toxic effects on bone marrow cells, and epigenetic modification is believed to be a potential mechanism underlying benzene pathogenesis. However, the specific roles of N6-methyladenosine (m6A), a newly discovered RNA post-transcriptional modification, in benzene-induced hematotoxicity remain unclear. In this study, we identified self-renewing malignant proliferating cells in the bone marrow of benzene-exposed mice through in vivo bone marrow transplantation experiments and Competitive Repopulation Assay. Subsequent analysis using whole transcriptome sequencing and RNA m6A methylation sequencing revealed a significant upregulation of RNA m6A modification levels in the benzene-exposed group. Moreover, RNA methyltransferase METTL14, known as a pivotal player in m6A modification, was found to be aberrantly overexpressed in Lin-Sca-1+c-Kit+ (LSK) cells of benzene-exposed mice. Further analysis based on the GEO database showed a positive correlation between the expression of METTL14, mTOR, and GFI and benzene exposure dose. In vitro cellular experiments, employing experiments such as western blot, q-PCR, m6A RIP, and CLIP, validated the regulatory role of METTL14 on mTOR and GFI1. Mechanistically, continuous damage inflicted by benzene exposure on bone marrow cells led to the overexpression of METTL14 in LSK cells, which, in turn, increased m6A modification on the target genes' (mTOR and GFI1) RNA. This upregulation of target gene expression activated signalling pathways such as mTOR-AKT, ultimately resulting in malignant proliferation of bone marrow cells. In conclusion, this study offers insights into potential early targets for benzene-induced haematologic malignant diseases and provides novel perspectives for more targeted preventive and therapeutic strategies.

Nervogenic Acid Derivatives and Phenanthrenes from Liparis nervosa and Their Antimicrobial and Immunosuppressive Activities.

Two new nervogenic acid derivatives liparisnervosides Q (1) and R (5), as well as five known nervogenic acid derivatives (2-4, 6, 7) and four phenanthrenes (8-11) were isolated from the whole plant of Liparis nervosa (Thunb. ex A. Murray) Lindl.. Their structures were detremined using extensive spectroscopic techniques, including 1D, 2D NMR, and HR-ESI-MS, and acid hydrolysis. Furthermore, their antimicrobial and immunosuppressive activities were evaluated. Nervosine VII (3) exhibited antimicrobial activity against Staphylococcus aureus with an MIC of 62.5 µg/mL and inhibited the proliferation of human T cells with an IC50 value of 9.67±0.96 µM. These findings contribute to our understanding of the potential pharmacological properties of these compounds.

A novel and independent survival prognostic model for OSCC: the functions and prognostic values of RNA-binding proteins.

BACKGROUND: Oral squamous cell carcinoma (OSCC), exhibiting high morbidity and malignancy, is the most common type of oral cancer. The abnormal expression of RNA-binding proteins (RBPs) plays important roles in the occurrence and progression of cancer. The objective of the present study was to establish a prognostic assessment model of RBPs and to evaluate the prognosis of OSCC patients. METHODS: Gene expression data in The Cancer Genome Atlas (TCGA) were analyzed by univariate Cox regression analysis model that established a novel nine RBPs, which were used to build a prognostic risk model. A multivariate Cox proportional regression model and the survival analysis were used to evaluate the prognostic risk model. Moreover, the receive operator curve (ROC) analysis was tested further the efficiency of prognostic risk model based on data from TCGA database and Gene Expression Omnibus (GEO). RESULTS: Nine RBPs' signatures (ACO1, G3BP1, NMD3, RNGTT, ZNF385A, SARS, CARS2, YARS and SMAD6) with prognostic value were identified in OSCC patients. Subsequently, the patients were further categorized into high-risk group and low-risk in the overall survival (OS) and disease-free survival (DFS), and external validation dataset. ROC analysis was significant for both the TCGA and GEO. Moreover, GSEA revealed that patients in the high-risk group significantly enriched in many critical pathways correlated with tumorigenesis than the low, including cell cycle, adheres junctions, oocyte meiosis, spliceosome, ERBB signaling pathway and ubiquitin-mediated proteolysis. CONCLUSIONS: Collectively, we developed and validated a novel robust nine RBPs for OSCC prognosis prediction. The nine RBPs could serve as an independent and reliable prognostic biomarker and guiding clinical therapy for OSCC patients.

Application of immersive virtual reality (IVR) for the care of common diseases of older adults course: A mixed methods study.

OBJECTIVE: This study aimed to enhance understanding, engagement, and learning efficiency in the course "The Care of Common Diseases of Older Adults" using a developed Immersive Virtual Reality(IVR) system. METHODS: A mixed-methods study with 32 students was conducted. The quantitative part involved a randomized controlled trial, and the qualitative part included thematic interviews with students and teachers. RESULTS: The intervention group using the IVR system showed significant improvements in positivity and performance evaluation scores (P < 0.05) compared to the control group. Negative affect scores also decreased significantly (P < 0.05). Qualitative data from interviews supported the quantitative findings, highlighting increased curiosity, learning enthusiasm, and academic performance. CONCLUSION: IVR significantly enhances learning by stimulating curiosity and active participation, making education more accessible and improving student performance. Future IVR enhancements should focus on user-friendliness and empathetic feedback in adult care.

E. urophylla × E. grandis high-quality genome and comparative genomics provide insights on evolution and diversification of eucalyptus.

BACKGROUND: Eucalyptus urophylla × Eucalyptus grandis, an economically important forest tree, provides important raw material for energy and reduces damage to native forests. However, the absence of a high-quality E. urophylla × E. grandis reference genome has significantly hindered its evolution and genetic analysis. RESULTS: We successfully presented a high-quality reference genome of E. urophylla × E. grandis (545.75 Mb; scaffold N50, 51.62 Mb) using a combination of the Illumina, PacBio HiFi, and Hi-C sequencing platforms. A total of 34,502 genes and 58.56% of the repetitive sequences in this genome were annotated. Using genome evolution analyses, we identified a recent whole-genome duplication (WGD) event in E. urophylla × E. grandis. We further found that gene families associated with starch and sucrose metabolism, flavonoid biosynthesis, and plant-pathogen interaction were significantly expanded in E. urophylla × E. grandis. Moreover, comparative genomic and evolutionary analyses showed large structural variations among the different chromosomes of the 34 Eucalyptus accessions, which were divided into six clades. CONCLUSIONS: Overall, our findings provide a valuable resource for expanding our understanding of the E. urophylla × E. grandis genome evolution, genetic improvement, and its comparative biology.

Transcriptome profiling reveals characteristics of hairy root and the role of AhGLK1 in response to drought stress and post-drought recovery in peanut.

BACKGROUND: HR (hairy root) has emerged as a valuable tissue for the rapid characterization of plant gene function and enzyme activity in vivo. AhGLK1 (Arachis hypogaea L. golden2-like 1) is known to play a role in post-drought recovery. However, it is unclear (a) whether HR has properties that are distinct from those of PR (primary root); and (b) which gene networks are regulated by AhGLK1 in response to drought stress and recovery in peanut. RESULTS: We found that cells of the root tip cortex were larger in HR than in PR, while a total of 850 differentially expressed genes (DEGs) were identified in HR compared to PR. Eighty-eight of these DEGs, relating to chlorophyll and photosynthesis, were upregulated in HR. In addition, AhGLK1-OX (AhGLK1-overexpressing) HR showed a green phenotype, and had a higher relative water content than 35 S::eGFP (control) HR during drought stress. RNA-seq analysis showed that 74 DEGs involved both in the drought response and the post-drought recovery process were significantly enriched in the galactose metabolism pathway. GO terms enrichment analysis revealed that 59.19%, 29.79% and 17.02% of the DEGs mapped to the 'biological process' (BP), 'molecular function' (MF) and 'cellular component' (CC) domains, respectively. Furthermore, 20 DEGs involved in post-drought recovery were uniquely expressed in AhGLK1-OX HR and were significantly enriched in the porphyrin metabolism pathway. GO analysis showed that 42.42%, 30.30% and 27.28% of DEGs could be assigned to the BP, MF and CC domains, respectively. Transcription factors including bHLH and MYB family members may play a key role during drought stress and recovery. CONCLUSION: Our data reveal that HR has some of the characteristics of leaves, indicating that HR is suitable for studying genes that are mainly expressed in leaves. The RNA-seq results are consistent with previous studies that show chlorophyll synthesis and photosynthesis to be critical for the role of AhGLK1 in improving post-drought recovery growth in peanut. These findings provide in-depth insights that will be of great utility for the exploration of candidate gene functions in relation to drought tolerance and/or post-drought recovery ability in peanut.

Burden of female infertility in China from 1990 to 2019: a temporal trend analysis and forecasting, and comparison with the global level.

BACKGROUND: Infertility is a common reproductive disease that affects not only individuals and families, but also the growth of the social population. Hence, understanding the burden of female infertility in China and worldwide is of great significance for the development of infertility prevention and treatment strategies. METHODS: The Global Burden of Disease Study (GBD 2019) Data Resources were used to collect and collate relevant data on female infertility in China and worldwide from 1990 to 2019. The difference in the number, age-standardised prevalence rate (ASPR), disability-adjusted life years and age-standardised disability-adjusted life years rate (ASDR) of women with infertility in different periods and geographical areas were analysed. The autoregressive integrated moving average method was used to predict the ASPR and ASDR of female infertility in China and worldwide in the next 11years. RESULTS: In the past 30years, the number of female infertility cases increased by 7.06million in China and 56.71million worldwide. The corresponding average annual increase of ASPR was 10.10% and 7.28%, respectively, and that of ASDR was 0.08% and 0.79%, respectively. In addition, there are differences in age and time between Chinese and global female infertility. In 1990, the crude prevalence rate of female infertility was the highest in women aged 40-44years and 35-39years in China and worldwide, respectively. In 2019, the crude prevalence rate of female infertility was still the highest in women aged 40-44years in China, whereas that around the world reached the highest in women aged 30-34years, which was significantly earlier. The forecast for the next 11years suggests that the ASPR and ASDR for female infertility in China will first rise and then decline, but the overall magnitude of change is not very significant, whereas the ASPR and ASDR for female infertility globally are still on the rise. The ASPR value of female infertility is expected to be 5025.56 in 100 000 persons in China and 3725.51 in 100 000 persons worldwide by 2030. The ASDR value of female infertility is expected to be 26.16 in 100 000 persons in China and 19.96 in 100 000 persons worldwide by 2030. CONCLUSION: The burden of female infertility is still increasing in China and worldwide. Therefore, it is of great significance to pay more attention to infertile women, and advocate a healthy lifestyle to reduce the burden of disease for infertile women.

First Report of Apple Brown Rot caused by Monilinia polystroma in Jilin Province in China.

Brown rot caused by Monilinia spp. is an important postharvest disease. It affects fruit quality and can cause serious economic losses. In October 2021, typical brown rot symptoms on fruit were observed at an apple orchard in Xiaobaishan Township, Jilin Province, China (E126°39'10″, N43°44'21″). Over 1200 plants were surveyed in the orchard, and nearly 25% of the plants were infected. In this research, samples from ten different trees showing typical symptoms were isolated and identified. Freshly diseased fruits were surface sterilized with 75% ethanol for 15s, then fungal colonies were isolated from 3 mm diameter diseased tissue samples. The purified colonies were placed on potato dextrose agar (PDA), oatmeal agar (OA) and water agar (WA + Sterilized apple pulp) and incubated at 25 ℃ in a 12 h/12 h light-dark photoperiod for 5 days. The colonies became light to dark brown; they grew faster on PDA with a growth rate of 5.53 mm/d, most densely on OA and slowest on SA + sterilized apple pulp with thin mycelia. After 20 days, some transparent, oval, smooth conidia were observed on the SA + sterile apple culture medium. Conidia sizes were 8.27-16.54 (avg. 11.97) x 2.92-7.09 (avg. 4.52) um (n=50) (Hilber-Bodmer et al. 2012). Pure cultures of eight samples were isolated from single spores, and DNA was extracted with a commercial nucleic acid extraction kit (Omega, cat#D3390-01). Then, fragments of the internal transcribed spacer region (ITS), translation elongation factor 1 alpha (EF1-alpha), beta-tubulin (Bt), and glyceraldehyde-3-phosphate dehydrogenase genes(G3pdh) were PCR amplified using primes ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999), BT-2a/BT-2b (Glass and Donaldson 1995), and G3pdhF/G3pdhR (Hu et al. 2011) respectively. The amplicons were sequenced by Takara Bio Inc. Sequences of all isolates were identical, thus one set of sequences were run with BLASTn against the NCBI GenBank nr database. The homology analysis showed that ITS (OQ170786), Bt (OQ179019), EF (OQ834046) and G3pdh (OQ834047) gene fragments were 100% (0/540 nt) , 100% (0/462 nt), 99.01% (3/304nt) and 99.61% (3/767 nt) similar to M. polystroma （NR154198.1, MT038414.1, LT632542.1, MT038415.1）respectively. Based on morphological characteristics and sequence analysis, the fungal isolate was identified as M. polystroma. Koch's postulates were conducted using ten healthy apple fruits that were surface disinfected with 0.2% sodium hypochlorite and repeatedly rinsed with sterile water. The test apples were wounded with a sterile needle, inoculated with mycelial agar plugs (3 mm diameter) on the wounds, and incubated at 25℃, 50% room humidity. The equivalent sterile PDA plugs were used as control and the experiment was repeated three times. After 5 days, brown rot symptoms appeared on the inoculated apples. 10 days later, nearly 1/3 of the inoculated apple surface appeared rotted, but the controls were symptomless. Subsequently, the same strain was re-isolated from the inoculated apples by pure culture and molecular identification. Therefore, M. polystroma (named JL-1) was confirmed as the causal agent of brown rot in Jilin Provincen China. M. polystroma is a typical pathogen of brown rot in the north of China, and only reported on apples in Shandong, apricots in Heilongjiang and pears in Hebei in China (Zhu et al. 2016) but never in Jilin. In addition, it was reported that the contribution of M. polystroma species to brown rot disease on apple and pear in China is 20% out of all the Monilinia spp. species that cause the disease, but M. polystroma virulence is not significantly different from other Monilinia species more widely distributed (Zhu et al, 2016). This is the first report of M. polystroma causing apple brown rot in Jilin Province of China. This finding will provide useful information for future diagnosis and management.

Comparison of maxillary anterior mathematical proportions among 3 dental arch forms.

STATEMENT OF PROBLEM: Race and sex differences in the mathematical proportions of maxillary anterior teeth have been evaluated. However, studies exploring the relationship between the mathematical proportion of maxillary anterior teeth and maxillary dental arch form are lacking. PURPOSE: The purpose of this clinical study was to determine if a correlation exists between the mathematical proportions of maxillary anterior teeth and 3 dental arch forms. MATERIAL AND METHODS: Three hundred young participants were recruited according to predefined criteria, and images and maxillary casts were obtained. The perceived width and length of the maxillary anterior teeth were measured on the images by using a software program, and mathematical proportions, including width ratios and width-to-length ratios, were calculated for each. The casts were used to categorize each specimen into tapered, ovoid, or square groups by using a specific classification method, and the width ratios and width-to-height ratios of the 3 dental arch forms were compared. Statistical analysis was performed by using analysis of variance (ANOVA) or the Kruskal-Wallis H test to compare the mathematical proportions of maxillary anterior teeth among the 3 dental arch forms (α=.05). The intraclass correlation coefficient (ICC) was used to test the reliability of the investigators. RESULTS: Except for the width-to-length ratios of the central incisor, mathematical proportions were affected by the dental arch form. The perceived width ratios of the maxillary lateral incisor-to-central incisor gradually increased from the tapered arch to the ovoid arch and from the ovoid arch to the square arch, with mean ±standard deviation values of 0.71 ±0.04, 0.73 ±0.05, and 0.79 ±0.06, respectively. For the maxillary perceived width ratios of canine-to-lateral incisor, the ovoid arch had the maximum perceived width ratios (0.86 ±0.10), followed by the tapered (0.82 ±0.10) and square arches (0.77 ±0.11). The width-to-length ratios of the central incisor were not affected by the arch form (P=.075), and the width-to-length ratios of the lateral incisor increased as the arch form became flat, with respective values of 0.70 ±0.10, 0.74 ±0.10, and 0.76 ±0.10. For the width-to-length ratios of the canine, the ovoid arch had the maximum perceived width ratios (0.58 ±0.10), followed by the tapered (0.53 ±0.10) and square arches (0.52 ±0.10). CONCLUSIONS: The mathematical proportions varied among the 3 dental arch forms, and the dental arch form should be considered during the prosthodontic design of maxillary anterior teeth.

Photo-Activated Nanofibrous Membrane with Self-Rechargeable Antibacterial Function for Stubborn Infected Cutaneous Regeneration.

For quick disinfection treatment, phototherapy, including photothermal therapy and photodynamic therapy, has emerged as a promising alternative to conventional methods. However, the bactericidal effect of phototherapy, which only works upon light, is short-lived. The remaining bacteria in situ may repopulate when the irradiation of light is withdrawn. To address this refractory concern, an antibacterial fibrous membrane consisting of electrospun poly (polycaprolactone) scaffolds and polydopamine (pDA) coated MXene/Ag3 PO4 bioheterojunctions (MX@AgP bio-HJs) is devised and developed. Upon near-infrared (NIR) illumination, the MX@AgP nanoparticle (NP) in nanofibrous electrospun membranes exert the excellent bactericidal effect of phototherapy and release Ag+ ions which stop the remaining bacteria from multiplying in the dark state. When removing NIR light, pDA in situ reduces Ag+ ions to Ag0 NPs to realize the self-rechargeability of Ag+ ions and provides enough Ag+ ions for the second phototherapy. In vivo results show that photoactivated nanofibrous membranes can re-shape an infected wound microenvironment to the regenerative microenvironment through killing bacteria, ceasing bleeding, increasing epithelialization, and collagen deposition on the wound bed, as well as promoting angiogenesis. As predicted, the proposal work offers potential prospects for nanofibrous membranes with NIR-assisted "self-rechargeable" antibacterial properties to treat bacteria-infected full-thickness wounds.

Systematical Ingredient Investigations of Ficus tikoua Bur. Fruit and Immunoregulatory and Antioxidant Effects of Different Fractions.

Although the fruit of Ficus tikoua Bur. has been consumed by montanic people in China for centuries, its chemical and biological composition was still unclear. A series of comprehensive investigations on its chemical constituents and bioactivities were carried out for the first time. As a result, six compounds were isolated and identified as the main components in this fruit. GC-MS analysis of the lipid components demonstrated that Ficus tikoua Bur. fruit contains some wholesome constituents such as fatty acids, vitamins, triterpenoids, and phytosterols. The fatty acids are mainly composed of linolenic acid (61.27%) and linoleic acid (22.79%). Furthermore, this fruit contains a relative high content of crude protein (9.41 ± 0.03%), total amino acids (9.28%), and total polyphenols (0.86 ± 0.01 g/100 g). The analysis of monosaccharide composition showed that the total polysaccharide mainly consists of glucose, glucuronic acid, xylose, arabinose, mannose, galactose, galacturonic acid, and rhamnose. The polysaccharide, polyphenol, water, ethanol, and flavonoid extracts exhibited prominent antioxidant activity determined by ABTS, DPPH, and FRAPS methods. Meanwhile, the total polysaccharide exhibited significant immunomodulatory effect by enhancing the release of cytokines and expression of iNOS and COX-2 in RAW264.7 cells, significantly decreasing the expression of c-Jun and p65 proteins in the cytoplasm; increasing the translocation of c-Jun and p65 to the nucleus; and regulating the phosphorylation level of Akt, PI3K, and PDK1 in the PI3K/AKT signaling pathway. This study proved that the fruit of F. tikoua is a reliable source of functional food.

Pseudolycorine chloride ameliorates Th17 cell-mediated central nervous system autoimmunity by restraining myeloid-derived suppressor cell expansion.

CONTEXT: The alkaloids of Narcissus tazetta L. var. Chinensis Roem (Amaryllidaceae) have antitumor and antiviral activities. However, the immunopharmacological effects of one of its constituents, pseudolycorine chloride (PLY), have not been reported yet. OBJECTIVE: We evaluated the effect of PLY on myeloid-derived suppressor cells (MDSCs) expansion and differentiation into monocyte-like MDSCs (M-MDSCs) and examined whether PLY alleviates Th17 cell-mediated experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis (MS). MATERIALS AND METHODS: In vitro, MDSCs were treated with PLY (0.67, 2 and 6 µM) or solcitinib (10 µM, positive control) for 48 or 96 h, and their proliferation, expansion, and differentiation into M-MDSCs were examined by flow cytometry. Myelin oligodendrocyte glycoprotein (MOG35-55) was used to induce EAE in female C57BL/6 mice, and the mice were treated with 40 mg/kg/d PLY or 1 mg/kg/d FK-506 (tacrolimus, positive control) for 21 days. Inflammatory infiltration, spinal cord demyelination, and MDSCs and Th17 cells infiltration into the spinal cord were examined using haematoxylin and eosin staining, Luxol fast blue staining, and immunofluorescence, respectively. RESULTS: In vitro, PLY (IC50/24 h = 6.18 µM) significantly inhibited IL-6 and GM-CSF-induced MDSCs proliferation, expansion and differentiation into M-MDSCs at all concentrations used. However, these concentrations did not show cytotoxicity. In mice, PLY (40 mg/kg) treatment alleviated EAE and inhibited inflammatory infiltration, demyelination, and MDSCs and Th17 cells infiltration into the spinal cord. DISCUSSION AND CONCLUSIONS: PLY may be an excellent candidate for the treatment of MS and other autoimmune diseases.

Indigoidine biosynthesis triggered by the heavy metal-responsive transcription regulator: a visual whole-cell biosensor.

During the last few decades, whole-cell biosensors have attracted increasing attention for their enormous potential in monitoring bioavailable heavy metal contaminations in the ecosystem. Visual and measurable output signals by employing natural pigments have been demonstrated to offer another potential choice to indicate the existence of bioavailable heavy metals in recent years. The biosynthesis of the blue pigment indigoidine has been achieved in E. coli following heterologous expression of both BpsA (a single-module non-ribosomal peptide synthetase) and PcpS (a PPTase to activate apo-BpsA). Moreover, we demonstrated herein the development of the indigoidine-based whole-cell biosensors to detect bioavailable Hg(II) and Pb(II) in water samples by employing metal-responsive transcriptional regulator MerR and PbrR as the sensory elements, and the indigoidine biosynthesis gene cluster as a reporter element. The resulting indigoidine-based biosensors presented a good selectivity and high sensitivity to target metal ions. High concentration of target metal exposure could be clearly recognized by the naked eye due to the color change by the secretion of indigoidine, and quantified by measuring the absorbance of the culture supernatants at 600 nm. Dose-response relationships existed between the exposure concentrations of target heavy metals and the production of indigoidine. Although fairly good linear relationships were obtained in a relatively limited concentration range of the concentrations of heavy metal ions, these findings suggest that genetically controlled indigoidine biosynthesis triggered by the MerR family transcriptional regulator can enable a sensitive, visual, and qualitative whole-cell biosensor for bioindicating the presence of bioaccessible heavy metal in environmental water samples. KEY POINTS: • Biosynthesis pathway of indigoidine reconstructed in a high copy number plasmid in E. coli. • Visual and colorimetric detection of Hg(II) and Pb(II) by manipulation of indigoidine biosynthesis through MerR family metalloregulator. •Enhanced detection sensitivity toward Hg(II) and Pb(II) achieved using novel pigment-based whole-cell biosensors.

Mechanically Strong, Liquid-Resistant Photothermal Bioplastic Constructed from Cellulose and Metal-Organic Framework for Light-Driven Mechanical Motion.

The development of photothermal materials with a high light-to-heat conversion capability is essential for the utilization of clean solar energy. In this work, we demonstrate the use of a novel and sustainable concept involving cellulose liquefaction, rapid gelation, in situ synthesis and hot-press drying to convert cellulose and metal-organic framework (Prussian blue) into a stable photothermal bioplastic that can harvest sunlight and convert it into mechanical motion. As expected, the obtained Prussian blue@cellulose bioplastic (PCBP) can effectively absorb sunlight and the surface can be heated up to 70.3 °C under one sun irradiation (100 mW cm-2). As a demonstration of the practicality of PCBP, it was successfully used to drive a Stirling engine motion. Meanwhile, hot-pressing promotes the densification of the structure of PCBP and, therefore, improves the resistance to the penetration of water/non-aqueous liquids. Moreover, PCBP shows good mechanical properties and thermal stability. Given the excellent photothermal performance and environmentally friendly features of photothermal conversion bioplastic, we envisage this sustainable plastic film could play important roles toward diversified applications: a photothermal layer for thermoelectric generator, agricultural films for soil mulching and photothermal antibacterial activity, among others.

Contact tracing of syphilis-seropositive pregnant women and syphilis-infection among their male partners in Bao'an district, Shenzhen, China.

BACKGROUND: Untreated male partners are a critical source of maternal re-infection. Contact tracing is a good way to identify infection among partners and reduce risk of mother-to-child transmission related to maternal re-infection. This study aimed to analyze the current situation and related factors of contact tracing of syphilis-seropositive pregnant women and syphilis-infection among their male partners. METHOD: Data of syphilis-seropositive pregnant women and their male partners attending clinic for syphilis-screening were obtained from the Shenzhen Program for Prevention of Congenital Syphilis. Contact tracing rate of syphilis-seropositive pregnant women and syphilis prevalence among male partners were counted, and related factors were also analyzed using a random-effects logistic regression model. RESULT: Of the 1299 syphilis-seropositive pregnant women, 74.1% (963/1299) had their male partners receiving syphilis-screening and 19.1% (184/963) of male partners were syphilis-infected. For pregnant women, being divorced (adjusted odds ratio [AOR] =0.39; 95%CI: 0.17-0.87), seeking for emergency services at their first antenatal clinics visits (AOR = 0.58; 95%CI: 0.44-0.77), reporting willingness to notify partner(AOR = 7.65; 95%CI: 4.69-12.49), multi-partners (AOR = 1.38; 95%CI:1.03-1.86) and having a history of drug abuse (AOR = 0.37; 95%CI: 0.14-1.00)were independently associated with successful contact tracing. For male partners, of minority ethnicity (AOR = 4.15; 95%CI: 1.66-10.34), age at first sex>20(AOR = 0.57; 95%CI: 0.37-0.87), reporting multi-partners (AOR = 1.60; 95%CI: 1.04-2.46), having a history of drug abuse (AOR = 4.07; 95%CI: 1.31-12.64) were independently associated with syphilis-infection. In addition, pregnant women with TRUST titer ≥1:8 (AOR = 2.81; 95%CI: 1.87-4.21), having a history of adverse pregnancy outcomes (AOR = 1.70; 95%CI: 1.14-2.53), reporting multi-partners (AOR = 0.43; 95%CI: 0.29-0.64) and reporting the current partner as the source of syphilis (AOR = 5.05; 95%CI: 2.82-9.03) were independently associated with partners' syphilis-infection. CONCLUSION: Contact tracing is feasible and effective in identifying syphilis-infected partners among syphilis-seropositive pregnant women. Contact tracing is associated with many factors such as women's marital status, services at their first antenatal clinics visit and willingness of partner notification. Partners' ethnicity, age at first sex, multi-partners and history of drug abuse as well as women's levels of TRUST titer were associated with partners' syphilis-infection.

Correction to: An efficient transgene-free DNA-editing system for Arabidopsis using a fluorescent marker.

The article An efficient transgene-free DNA-editing system for Arabidopsis using a fluorescent marker, written by Lejun Ouyang . Mingsai Ma and Limei Li, was originally published electronically on the publisher's internet portal (currently SpringerLink) on 03 December 2019 with open access. With the author(s)' decision to step back from Open Choice, the copyright of the article changed on 18 December 2019 to © Springer Nature B.V. 2019 and the article is forthwith distributed under the terms of copyright. The original article has been corrected.

An efficient transgene-free DNA-editing system for Arabidopsis using a fluorescent marker.

OBJECTIVE: To obtain transgene-free progeny by constructing a DNA editing system with a fluorescent screening marker gene and two pairs of single-guide RNAs to simultaneously recognize two different sites in the target gene encoding Arabidopsis microRNA(miR)160A RESULTS: The T1 seeds with red fluorescence were easily identified and were selected to verify that the proportion of miR160A knockout mutants reached approximately 50%. Seeds with no fluorescence in the T2 generation were selected and screened for homozygous mutants. In the T2 generation plants, the Cas9 fragment was not detected by polymerase chain reaction. The traits of the homozygous mutants were stably inherited by the T2 population. CONCLUSIONS: A highly efficient DNA-editing construct was successfully developed and can be used as a plant genome site-specific editing tool that may be useful for improving plant genetic resources.

Pichia kudriavzevii retards fungal decay by influencing the fungal community succession during cherry tomato fruit storage.

In this study, P. kudriavzevii was isolated and identified as an effective antagonistic yeast, which could significantly inhibit the rotting rate, weight loss, and delay the color change, with no effect on total soluble solids (TSS), titratable acid (TA), or firmness during cherry tomato storage. High-throughput sequencing was used to survey the effect of P. kudriavzevii on fungal community throughout cold storage. The results showed that the biological succession of predominant pathogens was disrupted by P. kudriavzevii. The abundance of Botrytis and Alternaria was higher in the control than upon P. kudriavzevii treatment at 28 d, but some yeast genera such as Naganishia, Wickerhamomyces, and Cutaneotrichosporon at 14 d, Pichia and Sporidiobolus at 21 d, and Cystofilobasidium at 28 d, had relatively higher abundances in P. kudriavzevii treatments than the control. Oddly, as an antagonist agent, P. kudriavzevii was not the dominant population, indicating that altering the course of succession of the fungal community may be an effective mechanism of antagonistic yeast. Furthermore, the total network correlation analysis of fungal community revealed that the community development was more dependent on similarities in function than on taxonomic relationships.

A benzoxazole derivative PO-296 inhibits T lymphocyte proliferation by the JAK3/STAT5 signal pathway.

Immunosuppressants have shown striking achievements in treating autoimmune diseases in recent years. It is urgent to develop more immunosuppressants to provide more options for patients. PO-296 [2-(6-chlorobenzo[d]oxazol-2-yl)-4,5,6,7-tetrahydro-2H-indazol-3-ol] was identified as a novel benzoxazole derivative. We observed that it exhibits an obvious immunosuppressive activity to T lymphocytes. PO-296 significantly inhibited the proliferation of activated human T lymphocyte without cytotoxicity. Moreover, PO-296 did not affect the expression of cluster of differentiation (CD)-25 or CD69 but induced T lymphocyte cycle arrest in the G0/G1 phase. Furthermore, PO-296 inhibited interleukin (IL)-6, IL-17, and interferon gamma expression but had no effect on IL-2, IL-4, or IL-10. Yet, importantly, PO-296 inhibited the phosphorylation of signal transducer and activator of transcription 5 (STAT5), increased the phosphorylation of p70S6K, but did not affect the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/mitogen-activated protein kinase pathway. In conclusion, these findings indicate that PO-296 inhibits human activated T-lymphocyte proliferation by affecting the janus kinase 3 (JAK3)/STAT5 pathway. PO-296 possesses a potential lead compound for the design and development of new immunosuppressants for the treatment of autoimmune diseases.