Enhanced degradation and methane production of food waste anaerobic digestate using an integrated system of anaerobic digestion and microbial electrolysis cells for long-term operation.

The integrated system of anaerobic digestion and microbial electrolysis cells (AD-MEC) was a novel approach to enhance the degradation of food waste anaerobic digestate and recover methane. Through long-term operation, the start-up method, organic loading, and methane production mechanism of the digestate have been investigated. At an organic loading rate of 4000 mg/L, AD-MEC increased methane production by 3-4 times and soluble chemical oxygen demand (SCOD) removal by 20.3% compared with anaerobic digestion (AD). The abundance of bacteria Fastidiosipila and Geobacter, which participated in the acid degradation and direct electron transfer in the AD-MEC, increased dramatically compared to that in the AD. The dominant methanogenic archaea in the AD-MEC and AD were Methanobacterium (44.4-56.3%) and Methanocalculus (70.05%), respectively. Geobacter and Methanobacterium were dominant in the AD-MEC by direct electron transfer of organic matter into synthetic methane intermediates. AD-MEC showed a perfect SCOD removal efficiency of the digestate, while methane as clean energy was obtained. Therefore, AD-MEC was a promising technology for deep energy transformation from digestate.

Biogas upgrading performance and underlying mechanism in microbial electrolysis cell and anaerobic digestion integrated system.

The productivity and efficiency of two-chamber microbial electrolysis cell and anaerobic digestion integrated system (MEC-AD) were promoted by a complex of anaerobic granular sludge and iron oxides (Fe-AnGS) as inoculum. Results showed that MEC-AD with Fe-AnGS achieved biogas upgrading with a 23%-29% increase in the energy recovery rate of external circuit current and a 26%-31% decrease in volatile fatty acids. The energy recovery rate of MEC-AD remained at 52%-57%, indicating a stable operation performance. The selectively enriched methanogens and electroactive bacteria resulted in dominant hydrogenotrophic and acetoclastic methanogenesis in the cathode and anode chambers. Mechanistic analysis revealed that MEC-AD with Fe-AnGS led to specifically upregulated enzymes related to energy metabolism and electron transfer. Fe-AnGS as inoculum could improve the long-term operation performance of MEC-AD. Consequently, this study provides an efficient strategy for biogas upgrading in MEC-AD.

Potential mechanisms of propionate degradation and methanogenesis in anaerobic digestion coupled with microbial electrolysis cell system: Importance of biocathode.

Microbial electrolysis cells (MEC) have the potential for enhancing the efficiency of anaerobic digestion (AD). In this study, microbiological and metabolic pathways in the biocathode of anaerobic digestion coupled with microbial electrolysis cells system (AD-MEC) were revealed to separate bioanode. The biocathode efficiently degraded 90 % propionate within 48 h, leading to a methane production rate of 3222 mL·m-2·d-1. The protein and heme-rich cathodic biofilm enhanced redox capacity and facilitated interspecies electron transfer. Key acid-degrading bacteria, including Dechloromonas agitata, Ignavibacteriales bacterium UTCHB2, and Syntrophobacter fumaroxidans, along with functional proteins such as cytochrome c and e-pili, established mutualistic relationships with Methanothrix soehngenii. This synergy facilitated a multi-pathway metabolic process that converted acetate and CO2 into methane. The study sheds light on the intricate microbial dynamics within the biocathode, suggesting promising prospects for the scalable integration of AD-MEC and its potential in sustainable energy production.

NSCLC extracellular vesicles containing miR-374a-5p promote leptomeningeal metastasis by influencing blood‒brain barrier permeability.

Leptomeningeal metastasis (LM) is a devastating complication of advanced non-small cell lung cancer (NSCLC). Diagnosis and monitoring of LM can be challenging. Extracellular vesicles (EVs) microRNAs (miRNAs) have become a new noninvasive diagnostic biomarker. The purpose of this study was to examine the clinical value and role of EVs miRNAs in NSCLC-LM. According to next-generation sequencing (NGS), miRNAs with differential expression of EVs in serum of NSCLC patients with LM and non-LM were detected to find biological markers for the diagnosis of LM. Cellular and in vivo experiments were conducted to explore the pathogenesis of EVs miRNA promoting LM in NSCLC. In the present study, we first demonstrated the serum level of EV-associated miR-374a-5p in patients with LM of lung cancer was much higher than that in patients without LM and was correlated with the survival time of patients with LM. Further studies showed that EVs miR-374a-5p efficiently destroys tight junctions and the integrity of the cerebral microvascular endothelial cell barrier, resulting in increased blood-brain barrier (BBB) permeability. Mechanistically, miR-374a-5p regulates the distribution of ZO-1 and occludin in endothelial cells by targeting ADD3, increasing vascular permeability and promoting LM. Implications: These results suggest that serum NSCLC-derived EVs miR-374a-5p is involved in premetastatic niche formation by regulating the permeability of BBB to promote NSCLC-LM, and can be used as a blood biomarker for the diagnosis and prognosis of NSCLC-LM.

Enhancing CH4 production in microbial electrolysis cells: Optimizing electric field via carbon cathode resistivity.

Microbial electrolysis cells (MECs) are increasingly recognized as a promising technology for converting CO2 to CH4, offering the dual benefits of energy recovery from organic wastewater and CO2 emission reduction. A critical aspect of this technology is the enhancement of the electron-accepting capacity of the methanogenic biocathode to improve CH4 production efficiency. This study demonstrates that adjusting the cathode resistivity is an effective way to control the electric field intensity, thereby enhancing the electron accepting capacity and CH4 production. By maintaining the electric field intensity within approximately 8.50-10.83 mV·cm-1, the CH4 yield was observed to increase by up to two-fold. The improvement in CH4 production under optimized electric field conditions was attributed to the enhancement of the direct accepting capacity of the biocathode. This enhancement was primarily due to an increase in the relative abundance of Methanosaeta by approximately 10 % and an up to 83.78 % rise in the electron-accepting capacity of the extracellular polymeric substance. These insights offer a new perspective on the operation of methanogenic biocathodes and propose a novel biocathode construction methodology based on these findings, thus contributing to the enhancement of MEC efficiency.

The influencing mechanism of AD-MEC domesticated sludge to alleviates propionate accumulation and enhances methanogenesis.

Anaerobic digestion combined with microbial electrolysis cell (AD-MEC) could maintain stable reactor operation and alleviating the anaerobic digestion (AD) propionate accumulation. In this study, the addition of sludge to AD-MEC was examined as a way to enhance system performance and explore the microbial interaction mechanism after electric field domestication. The results showed that under 1000 and 4000 mg/L propionate, the methane production of the sludge from AD-MEC increased by 34.29 % and 9.70 %, respectively, as compared to the AD sludge. Gompertz fitting analysis showed that sludge after electric field domestication enhancing its continuous methanogenic capacity. Further analysis showed that sludge extracellular electron transfer capacity was enhanced in AD-MEC and that its domesticated granular sludge formed a microbial community function with acid-degrading synergistic methanogenesis. The results of the study may provide theoretical support and optimization strategies for the application of AD-MEC system.

Cerebrospinal fluid circulating tumour DNA genotyping and survival analysis in lung adenocarcinoma with leptomeningeal metastases.

PURPOSE: The prognosis of patients with leptomeningeal metastasis (LM) remains poor. Circulating tumour DNA (ctDNA) has been proven to be abundantly present in cerebrospinal fluid (CSF); hence, its clinical implication as a biomarker needs to be further verified. METHODS: We conducted a retrospective study of 35 lung adenocarcinoma (LUAD) patients with LM, and matched CSF and plasma samples were collected from all patients. All paired samples underwent next-generation sequencing (NGS) of 139 lung cancer-associated genes. The clinical characteristics and genetic profiling of LM were analysed in association with survival prognosis. RESULTS: LM showed genetic heterogeneity, in which CSF had a higher detection rate of ctDNA (P = 0.003), a higher median mutation count (P < 0.0001), a higher frequency of driver mutations (P < 0.01), and more copy number variation (CNV) alterations (P < 0.001) than plasma. The mutation frequencies of the EGFR, TP53, CDKN2A, MYC and CDKN2B genes were easier to detect in CSF than in LUAD tissue (P < 0.05), possibly reflecting the underlying mechanism of LM metastasis. CSF ctDNA is helpful for analysing the mechanism of EGFR-TKI resistance. In cohort 1, which comprised patients who received 1/2 EGFR-TKIs before the diagnosis of LM, TP53 and CDKN2A were the most common EGFR-independent resistant mutations. In cohort 2, comprising those who progressed after osimertinib and developed LM, 7 patients (43.75%) had EGFR CNV detected in CSF but not plasma. Furthermore, patient characteristics and various genes were included for interactive survival analysis. Patients with EGFR-mutated LUAD (P = 0.042) had a higher median OS, and CSF ctDNA mutation with TERT (P = 0.013) indicated a lower median OS. Last, we reported an LM case in which CSF ctDNA dynamic changes were well correlated with clinical treatment. CONCLUSIONS: CSF ctDNA could provide a more comprehensive genetic landscape of LM, indicating the potential metastasis-related and EGFR-TKI resistance mechanisms of LM patients. In addition, genotyping of CSF combined with clinical outcomes can predict the prognosis of LUAD patients with LM.

Cerebrospinal fluid exosomal protein alterations via proteomic analysis of NSCLC with leptomeningeal carcinomatosis.

PURPOSE: Leptomeningeal carcinomatosis (LC) is a rare complication of non-small cell lung cancer (NSCLC) with highly mortality. Cerebrospinal fluid (CSF) as a special kind of tumor microenvironment (TME) better represents alterations than plasma. However, the clinical value of protein profiles of exosome in CSF as liquid biopsy remains unclear. METHODS: In this study, CSF samples of NSCLC patients with (LC group) or without (NSCLC group) LC were collected and compared to patients without tumors (normal group). CSF exosomes were isolated by ultracentrifugation and protein profiles were performed by label-free proteomics. Differentially expressed proteins (DEPs) were detected by bioinformatics tools and verified by parallel reaction monitoring (PRM). RESULTS: A total of 814 proteins were detected. Bioinformatics analysis revealed their shared function in the complement activation, extracellular region, and complement and coagulation cascades. Between LC and NSCLC group, 72 DEPs were found among which FN1 demonstrated the highest betweenness centrality (BC) after protein-protein interaction network analysis. CONCLUSION: We investigated the application of label free and PRM based proteomics to detect key proteins related to LC. FN1 may serve as potential indicator to classify LC and NSCLC. Extracellular matrix (ECM) and epithelial-mesenchymal transition (EMT) are important in the process of LC. These data is promising for early prediction and diagnosis of LC.

Next-generation sequencing in the diagnosis of neurobrucellosis: a case series of eight consecutive patients.

BACKGROUND: Neurobrucellosis (NB) presents a challenge for rapid and specific diagnosis. Next-generation sequencing (NGS) of cerebrospinal fluid (CSF) has showed power in detection of causative pathogens, even some infrequent and unexpected pathogens. In this study, we presented 8 cases of NB diagnosed by the NGS of CSF. METHODS: Between August 1, 2018 and September 30, 2020, NGS was used to detect causative pathogens in clinically suspected central nervous system (CNS) infections. Data on demographics, clinical features, and laboratory tests, imaging results and NGS results were collected and reviewed. RESULTS: Among the presented 8 patients, Brucella was rapidly detected using NGS of CSF within 1-4 days, despite those eight patients had variable medical history, disease course, clinical manifestations, laboratory tests and imaging findings. NGS showed the sequence reads corresponded to Brucella species were 8 to 448, with genomic coverage of 0.02 to 0.87%. The relative abundance was 0.13% to 82.40% and sequencing depth was 1.06 to 1.24. Consequently, patients were administered with 3 to 6 months of doxycycline, ceftriaxone and rifampicin, double or triple combination, supplemented with symptomatic therapy and were fully recovered except for case 1. CONCLUSION: NGS of CSF provides a powerful tool in detection of Brucella in a prompt and specific manner, and can be considered for first-line diagnostic use in practice.

Genomic alterations of cerebrospinal fluid cell-free DNA in leptomeningeal metastases of gastric cancer.

BACKGROUND: Leptomeningeal metastases (LM) were rare in gastric cancer (GC), and GC patients with LM (GCLM) generally suffer from poor prognosis. Nevertheless, the clinical utility of cerebrospinal fluid (CSF) circulating tumor DNA (ctDNA) was underinvestigated in GCLM. METHODS: We retrospectively studied 15 GCLM patients, and all patients had paired primary tumor tissue samples and post-LM CSF samples while 5 patients also had post-LM plasma samples. All samples were analyzed using next-generation sequencing (NGS), and the molecular and clinical features were correlated with clinical outcomes. RESULTS: CSF had higher mutation allele frequency (P = 0.015), more somatic mutations (P = 0.032), and more copy-number variations (P < 0.001) than tumor or plasma samples. Multiple genetic alterations and aberrant signal pathways were enriched in post-LM CSF, including CCNE1 amplification and cell cycle-related genes, and CCNE1 amplification was significantly associated with patients' overall survival (P = 0.0062). More potential LM progression-related markers were detected in CSF samples than in tumor samples, including PREX2 mutation (P = 0.014), IGF1R mutation (P = 0.034), AR mutation (P = 0.038), SMARCB1 deletion (P < 0.001), SMAD4 deletion (P = 0.0034), and TGF-beta pathway aberration (P = 0.0038). Additionally, improvement in intracranial pressure (P < 0.001), improvement in CSF cytology (P = 0.0038), and relatively low levels of CSF ctDNA (P = 0.0098) were significantly associated with better PFS. Lastly, we reported a GCLM case whose CSF ctDNA dynamic changes were well correlated with his clinical assessment. CONCLUSIONS: CSF ctDNA could more sensitively detect molecular markers and metastasis-related mechanisms than tumor tissues in GCLM patients, and our study sheds light on utilizing CSF ctDNA in prognostic estimation and clinical assessment in GCLM.

Next-generation sequencing for the diagnosis of Listeria monocytogenes meningoencephalitis: a case series of five consecutive patients.

Introduction. The prompt and specific diagnosis of Listeria monocytogenes meningoencephalitis (LMM) is challenging. Next-generation sequencing (NGS) of cerebrospinal fluid (CSF) is an emerging technique for diagnosing infrequent causative pathogens.Hypothesis/Gap statement. We hypothesized that NGS of CSF is an effective approach for diagnosing LMM.Aim. To evaluate the effectiveness of NGS, we present five cases of LMM diagnosed using NGS of the CSF.Methodology. Between August 2017 and 30 September 2020, we used NGS of the CSF to detect pathogens in patients with clinically suspected central nervous system infections. The clinical characteristics, laboratory tests, imaging findings and NGS results are reviewed.Results. Five patients were diagnosed with LMM using NGS of the CSF within 2 to 4 days, although the clinical manifestations, medical history and imaging findings varied strikingly. NGS of CSF showed sequence reads corresponding to L. monocytogenes species ranging from 118 to 1997 bp, genomic coverage of 0.29-5.96 %, relative abundance of 14.83-32.16 % and sequencing depth of 1.12 to 1.35. The prompt diagnosis resulted in targeted and effective treatment with the appropriate antibiotics, although two patients with the most severe cerebral parenchymal lesions showed little improvement.Conclusion. Our results demonstrate the power of NGS of CSF for the prompt diagnosis of LMM. NGS of CSF is an important complementary tool for identifying L. monocytogenes.

Combined Effects of Programmed Cell Death-1 Blockade and Endostar on Brain Metastases of Lung Cancer.

BACKGROUND: The blockade of programmed cell death-1 (PD-1) and recombinant human endostatin can be used for the treatment of non-small cell lung cancer (NSCLC) and its metastasis. This study aims to explore the therapeutically potential of PD-1 blockade plus Endostar in brain metastasis of NSCLC. METHODS: The mouse brain metastases model was established using Lewis lung carcinoma luciferase (LLC-Luc) and PC-9-Luc cells. Tumor metastasis in the brain and tumor burden were analyzed by using bioluminescence imaging (BLI), qRT-PCR and ELISA which were used to determine the mRNA and protein levels of biomarkers in tumor tissues. Immunohistochemical staining was used to determine the expression and location of CD31 in tumor tissues in the brain. RESULTS: Treatment with anti-PD-1 and Endostar suppressed tumor metastasis in the brain and prolonged overall survival rate in LLC-Luc and PC-9-Luc brain metastases mouse model. In addition, treatment with anti-PD-1 and Endostar inhibited the expressions of CD31 and VEGF in tumor tissues in the brain. Furthermore, treatment with anti-PD- 1 and Endostar significantly suppressed the levels of IL1ß, IFNγ, and TGFß in the tumor tissues. CONCLUSION: The combination of PD-1 blockade and endostar suppressed brain metastases of NSCLC.

Metagenomic next-generation sequencing of cell-free and whole-cell DNA in diagnosing central nervous system infections.

Background: Central nervous system (CNS) infections pose a fatal risk to patients. However, the limited sample volumes of cerebrospinal fluid (CSF) and low detection efficiency seriously hinder the accurate detection of pathogens using conventional methods. Methods: We evaluated the performance of metagenomics next-generation sequencing (mNGS) in diagnosing CNS infections. CSF samples from 390 patients clinically diagnosed with CNS infections were used for the mNGS of cell-free DNA (cfDNA) (n =394) and whole-cell DNA (wcDNA) (n =150). Results: The sensitivity of mNGS using cfDNA was 60.2% (237/394, 95% confidence interval [CI] 55.1%-65.0%), higher than that of mNGS using wcDNA (32.0%, 95% [CI] 24.8%-40.2%, 48/150) and conventional methods (20.9%, 95% [CI] 16.2%-26.5%, 54/258) (P < 0.01, respectively). The accuracy of mNGS using cfDNA in positive samples was 82.6%. Most of viral (72.6%) and mycobacterial (68.8%) pathogens were only detected by the mNGS of cfDNA. Meningitis and encephalitis with Streptococcus pneumoniae infection might be more likely to result in critically ill diseases, while Human alphaherpesvirus 3 was prone to cause non-critically ill diseases. Conclusions: This is the first report on evaluating and emphasizing the importance of mNGS using CSF cfDNA in diagnosing CNS infections, and its extensive application in diagnosing CNS infections could be expected, especially for viral and mycobacterial CNS infections.

Effects of Biodegradation of Corn-Starch-Sodium-Alginate-Based Liquid Mulch Film on Soil Microbial Functions.

In response to the problems of the poor degradability and mechanical properties of liquid mulch, natural non-toxic polymer compound corn starch and sodium alginate were used to prepare fully biodegradable liquid mulch. The preparation conditions of the mulch were optimized, and the mechanical properties of the mulch and the changes in the microbial community in soil with the mulch degradation were analyzed. The corn-starch-sodium-alginate-based liquid mulch film had an optimum performance at a tensile strength of 0.145 MPa and an elongation at a break of 16.05%, which was attained by adding 33.33% sodium alginate, 50% glycerol 22 and 4% citric acid to corn starch after moist heat modification. Fourier transform infrared spectroscopy analysis showed that the -COOH in sodium alginate could interact with the -OH in starch and glycerol through hydrogen bonding, thus, resulting in a denser structure and better mechanical properties of the liquid mulch as a non-crystalline material. The soil burial degradation study of mulch revealed that corn-starch-sodium-alginate-based liquid mulch degraded completely at 25 days macroscopically, and mulch degradation increased soil organic matter content. Microbial kinetic analysis showed that the abundance and diversity of the bacterial community decreased with the degradation of the mulch, which was conducive to the optimization of the bacterial community structure and function. Arthrobacter of the class Actinomycetes became the dominant microorganism, and its abundance increased by 16.48-times at 14 days of mulch degradation compared with that before degradation, and Acidophilus phylum (14 days) decreased by 99.33%. The abundance of fungal communities was elevated in relation to the main functional microorganisms involved in liquid mulch degradation, with Alternaria and Cladosporium of the Ascomycete phylum Zygomycetes being the most active at the early stage of mulch degradation (7 days), and the relative abundance of Blastocystis was significantly elevated at the late stage of mulch degradation (14 days), which increased by 13.32%. This study provides important support for the green and sustainable development of modern agriculture.

A Promising Approach to Mitigate Risks on the Existing Tailings Dams in Brazil.

Mining operations lead to the production of large quantities of mineral waste, such as fluid fine tailings whose disposal is rather challenging. In Brazil, tailings disposal is facing a critical situation due to the large accidents that took place in the last couple of years. As a response to these accidents, the Brazilian Mining Agency became stricter on licensing mining complexes and issued an interruption on activities on 56 tailings dams. In this paper, the authors propose a promising approach to minimize risks on the existing tailings dams hoping that the industry succeeds at mining avoiding social harms and environmental damages. The results presented herein showed that in situ electrical dewatering is a promising technology that offers many benefits. It can significantly increase the solids content of liquefiable tailings stratum within a short time. This leads to tailings masses possessing higher shear strength. As a consequence, this technology might well lead to safe tailing dams in Brazil bringing socio-economic and environmental benefits.

Development of a rapid startup method of direct electron transfer-dominant methanogenic microbial electrosynthesis.

The rapid startup of carbon dioxide reduction-methanogenic microbial electrosynthesis is crucial for its industrial application, and the development of cathode biofilm is the key to its industrialization. Based on the new discovery that biofilm formed by placing graphite felt in an anaerobic reactor was electroactive, with strong direct electron transfer and methanogenesis ability (24.52 mL/L/d), a new startup method was developed. The startup time was shortened by at least 20 days and charge transfer resistance was reduced by 4.45-10.78 times than common startup methods (inoculating cathode effluent or granular sludge into the cathode chamber). The new method enriched electroactive bacteria. Methanobacterium and Methanosaeta accounted for 62.04% and 34.96%, respectively. The common methods inoculating cathode effluent or granular sludge enriched hydrogenotrophic microorganisms (>95%) or Methanosaeta (54.10%) due to the local environments of cathode. This new rapid and easy startup method may support the scale-up of microbial electrosynthesis.

Value of Neutrophil to Lymphocyte Ratio, Platelet to Lymphocyte Ratio, and Red Blood Cell Distribution Width in Evaluating the Prognosis of Children with Severe Pneumonia.

OBJECTIVE: To investigate the value of neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), and red blood cell distribution width (RDW) in evaluating the prognosis of children with severe pneumonia (SP). METHODS: A retrospective analysis of the data of 91 children with SP admitted to our hospital from March 2018 to March 2021. According to the survival status after 28 days of treatment, all children were divided into the survival group (n = 59) and the death group (n = 32). The clinical data and laboratory indicators of the patients were recorded. Multivariate logistic regression was used to analyze the risk factors of prognosis, and the ROC curve was used to analyze the predictive value of each index. RESULTS: The Acute Physiology and Chronic Health Evaluation II (APACHE II) score and CURB-65 score of the death group were higher than those of the survival group (P < 0.05). The RDW, NLR, PLR, and high-sensitivity C-reactive protein, procalcitonin blood lactic acid (Lac) of the death group, were higher than those of the survival group, and LYM was lower than the survival group (P < 0.05). Multivariate regression analysis showed that APACHE II score, RDW, NLR, PLR, and Lac were all independent risk factors for poor prognosis in children with SP (P < 0.05). The AUC of NLR, PLR, and RDW for evaluating the prognosis of children with SP were 0.798, 0.781, and 0.777, respectively. The sensitivity was 56.25%, 90.63%, and 56.25%, respectively, and the specificity was 89.83%, 55.93%, and 91.53%, respectively. The AUC of NLR, PLR, and RDW combined to evaluate the prognosis of children with SP was 0.943. When the best cut-off value was 0.8528, the sensitivity was 93.75%, and the specificity was 91.53%. CONCLUSION: NLR, PLR, and RDW have certain predictive value for the prognosis of children with SP; the combination of the three indicators has a higher value in evaluating the prognosis of children with SP, which can better guide the prognostic treatment.

Doxorubicin-loaded nanoparticle coated with endothelial cells-derived exosomes for immunogenic chemotherapy of glioblastoma.

Treatments of glioblastoma (GBM) have not been very effective, largely due to the inefficiency of drugs in penetrating the blood brain barrier (BBB). In this study, we investigated the potential of exosome-coated doxorubicin (DOX)-loaded nanoparticles (ENPDOX) in BBB penetration, inducing immunogenic cell death (ICD) and promoting survival of GBM-bearing mice. DOX-loaded nanoparticles (NPDOX) were coated with exosomes prepared from mouse brain endothelial bEnd.3 cells. ENPDOX cellular uptake was examined. Penetration of ENPDOX through the BBB was tested in an in vitro transwell system and a GBM mouse model. The effects of ENPDOX in inducing apoptosis and ICD were assessed. Finally, the efficacy of ENPDOX in the treatment of GBM-bearing mice was assessed. ENPDOX was taken up by bEnd.3 cells and could penetrate the BBB both in vitro and in vivo. In vitro, ENDDOX induced apoptosis and ICD of glioma GL261 cells. Systemic administration of ENPDOX resulted in maturation of dendritic cells, activation of cytotoxic cells, altered production of cytokines, suppressed proliferation and increased apoptosis of GBM cells in vivo and prolonged survival of GBM-bearing mice. Our findings indicate that ENPDOX may be a potent therapeutic strategy for GBM which warrants further investigation in clinical application.

Nitrate Modulates Lateral Root Formation by Regulating the Auxin Response and Transport in Rice.

Nitrate (NO3-) plays a pivotal role in stimulating lateral root (LR) formation and growth in plants. However, the role of NO3- in modulating rice LR formation and the signalling pathways involved in this process remain unclear. Phenotypic and genetic analyses of rice were used to explore the role of strigolactones (SLs) and auxin in NO3--modulated LR formation in rice. Compared with ammonium (NH4+), NO3- stimulated LR initiation due to higher short-term root IAA levels. However, this stimulation vanished after 7 d, and the LR density was reduced, in parallel with the auxin levels. Application of the exogenous auxin α-naphthylacetic acid to NH4+-treated rice plants promoted LR initiation to levels similar to those under NO3- at 7 d; conversely, the application of the SL analogue GR24 to NH4+-treated rice inhibited LR initiation to levels similar to those under NO3- supply by reducing the root auxin levels at 10 d. D10 and D14 mutations caused loss of sensitivity of the LR formation response to NO3-. The application of NO3- and GR24 downregulated the transcription of PIN-FORMED 2(PIN2), an auxin efflux carrier in roots. LR number and density in pin2 mutant lines were insensitive to NO3- treatment. These results indicate that NO3- modulates LR formation by affecting the auxin response and transport in rice, with the involvement of SLs.

SPL14/17 act downstream of strigolactone signalling to modulate rice root elongation in response to nitrate supply.

Nitrogen (N) is an essential major nutrient for food crops. Although ammonium (NH4+ ) is the primary N source of rice (Oryza sativa), nitrate (NO3- ) can also be absorbed and utilized. Rice responds to NO3- application by altering its root morphology, such as root elongation. Strigolactones (SLs) are important modulators of root length. However, the roles of SLs and their downstream genes in NO3- -induced root elongation remain unclear. Here, the levels of total N and SL (4-deoxyorobanchol) and the responses of seminal root (SR) lengths to NH4+ and NO3- were investigated in rice plants. NO3- promoted SR elongation, possibly due to short-term signal perception and long-term nutrient function. Compared with NH4+ conditions, higher SL signalling/levels and less D53 protein were recorded in roots of NO3- -treated rice plants. In contrast to wild-type plants, SR lengths of d mutants were less responsive to NO3- conditions, and application of rac-GR24 (SL analogue) restored SR length in d10 (SL biosynthesis mutant) but not in d3, d14, and d53 (SL-responsive mutants), suggesting that higher SL signalling/levels participate in NO3- -induced root elongation. D53 interacted with SPL17 and inhibited SPL17-mediated transactivation from the PIN1b promoter. Mutation of SPL14/17 and PIN1b caused insensitivity of the root elongation response to NO3- and rac-GR24 applications. Therefore, we conclude that perception of SLs by D14 leads to degradation of D53 via the proteasome system, which releases the suppression of SPL14/17-modulated transcription of PIN1b, resulting in root elongation under NO3- supply.