Enhanced organic degradation and microbial community cooperation by inoculating Bacillus licheniformis in low temperature composting.

Microbial activity and interaction are the important driving factors in the start-up phase of food waste composting at low temperature. The aim of this study was to explore the effect of inoculating Bacillus licheniformis on the degradation of organic components and the potential microbe-driven mechanism from the aspects of organic matter degradation, enzyme activity, microbial community interaction, and microbial metabolic function. The results showed that after inoculating B. licheniformis, temperature increased to 47.8°C on day 2, and the degradation of readily degraded carbohydrates (RDC) increased by 31.2%, and the bioheat production increased by 16.5%. There was an obvious enhancement of extracellular enzymes activities after inoculation, especially amylase activity, which increased by 7.68 times on day 4. The inoculated B. licheniformis colonized in composting as key genus in the start-up phase. Modular network analysis and Mantel test indicated that inoculation drove the cooperation between microbial network modules who were responsible for various organic components (RDC, lipid, protein, and lignocellulose) degradation in the start-up phase. Metabolic function prediction suggested that carbohydrate metabolisms including starch and sucrose metabolism, glycolysis / gluconeogenesis, pyruvate metabolism, etc., were improved by increasing the abundance of related functional genes after inoculation. In conclusion, inoculating B. licheniformis accelerated organic degradation by driving the cooperation between microbial network modules and enhancing microbial metabolism in the start-up phase of composting.

Promoting agricultural waste-driven denitrification and nitrogen sequestration with nano-enabled strategy.

Nanotechnology and biotechnology offer promising avenues for bolstering food security through the facilitation of soil nitrogen (N) sequestration and the reduction of nitrate leaching. Nonetheless, a comprehensive and mechanistic evaluation of their effectiveness and safety remains unclear. In this study, a soil remediation strategy employing nano-Fe3O4 and straw in N-contaminated soil was developed to elucidate N retention mechanisms via diverse metagenomics techniques. The findings revealed that subsoil amended with straw, particularly in conjunction with nano-Fe3O4, significantly increased subsoil N content (53.2%) and decreased nitrate concentration (74.6%) in leachate. Furthermore, the enrichment of functional genes associated with N-cycling, sulfate, nitrate, and iron uptake, along with chemotaxis, and responses to environmental stimuli or microbial collaboration, effectively mitigates nitrate leaching while enhancing soil N sequestration. This study introduces a pioneering approach utilizing nanomaterials in soil remediation, thereby offering the potential for the cultivation of safe vegetables in high N input greenhouse agriculture.

Effects of aeration modes and rates on nitrogen conversion and bacterial community in composting of dehydrated sludge and corn straw.

Aeration is an important factor to regulate composting efficiency and nitrogen loss. This study is aimed to compare the effects of different aeration modes (continuous and intermittent) and aeration rate on nitrogen conversion and bacterial community in composting from dehydrated sludge and corn straw. Results showed that the intermittent aeration mode at same aeration volume was superior to the continuous aeration mode in terms of NH3 emission reduction, nitrogen conversion and germination index (GI) improvement. Intermittent aeration mode with 1200 L/h (aeration 5 min, stop 15 min) [K5T15 (V1200)] and 300 L/h of continuous aeration helped to the conservation of nitrogen fractions and accelerate the composting process. However, it was most advantageous to use 150 L/h of continuous aeration to reduce NH3 emission and ensure the effective composting process. The aeration mode K5T15 (V1200) showed the fastest temperature rise, the longer duration of thermophilic stage and the highest GI (95%) in composting. The cumulative NH3 emission of intermittent aeration mode was higher than continuous aeration mode. The cumulative NH3 emission of V300 was 23.1% lower than that of K5T15 (V1200). The dominant phyla in dehydrated sludge and corn straw composting were Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes. The dominant phylum in the thermophilic stage was Firmicutes (49.39%~63.13%), and the dominant genus was Thermobifida (18.62%~30.16%). The relative abundance of Firmicutes was greater in the intermittent aeration mode (63.13%) than that in the continuous aeration mode (57.62%), and Pseudomonas was dominant in composting with lower aeration rate and the lowest NH3 emission. This study suggested that adjustment to the aeration mode and rate could affect core bacteria to reduce the nitrogen loss and accelerate composting process.

Phosphate additives promote humic acid carbon and nitrogen skeleton formation by regulating precursors and composting bacterial communities.

This study aimed to compare the effect of different phosphate additives including superphosphate (CP) and MP [Mg(OH)2 + H3PO4] on nitrogen conversion, humus fractions formation and bacterial community in food waste compost. The results showed the ratio of humic acid nitrogen in total nitrogen (HA-N/TN) in CP increased by 49 %. Ammonium nitrogen accumulation was increased by 75 % (CP) and 44 % (MP). Spectroscopic techniques proved that phosphate addition facilitated the formation of complex structures in HA. CP enhanced the dominance of Saccharomonospora, while Thermobifida and Bacillus were improved in MP. Structural equation modeling and network analysis demonstrated that ammonium nitrogen can be converted to HA-N and has positive effects on bacterial composition, reducing sugars and amino acids, especially in CP with more clustered network and synergic bacterial interactions. Therefore, the addition of phosphate provides a new idea to regulate the retained nitrogen toward humification in composting.

The contribution of microbial necromass to soil organic carbon and influencing factors along a variation of habitats in alpine ecosystems.

A growing consensus is reached that microbes contributes to regulating the formation and accumulation of soil organic carbon (SOC). Nevertheless, less is known about the role of soil microbes (necromass, biomass) in SOC accumulation in different habitat conditions in alpine ecosystems. To address this knowledge gap, the composition and distribution of amino sugars (ASs) and phospholipid fatty acids (PLFAs) as biomarkers of microbial necromass and biomass were investigated in forest, meadow and wetland soil profile (0-40 cm) of Mount Segrila, Tibet, China, as well the contribution of bacterial and fungal necromass to SOC. The results revealed that microbial necromass carbon contributed 45.15 %, 72.51 % and 78.08 % on average to SOC in 0-40 cm forest, meadow and wetland soils, respectively, and decreased with microbial biomass. Fungal necromass contributed more to SOC in these habitats than bacterial necromass. Microbial necromass increased with microbial biomass and both of them decreased with soil depth in all habitats. The necromass accumulation coefficient was significantly correlated with microbial necromass and biomass, affected by habitat and soil moisture. Structural equation model indicated that soil abiotic factors indirectly mediated the accumulation of SOC through microbial necromass and biomass. This study revealed that different habitats and soil depths control considerably soil physicochemical properties and microbial community, finally influencing SOC accumulation in alpine ecosystems, which emphasized the influence of abiotic factors on microbial necromass and biomass for SOC accumulation in alpine ecosystems.

Oxidative Stress Promotes Liver Cancer Metastasis via RNF25-Mediated E-Cadherin Protein Degradation.

Loss of E-cadherin (ECAD) is required in tumor metastasis. Protein degradation of ECAD in response to oxidative stress is found in metastasis of hepatocellular carcinoma (HCC) and is independent of transcriptional repression as usually known. Mechanistically, protein kinase A (PKA) senses oxidative stress by redox modification in its ß catalytic subunit (PRKACB) at Cys200 and Cys344. The activation of PKA kinase activity subsequently induces RNF25 phosphorylation at Ser450 to initiate RNF25-catalyzed degradation of ECAD. Functionally, RNF25 repression induces ECAD protein expression and inhibits HCC metastasis in vitro and in vivo. Altogether, these results indicate that RNF25 is a critical regulator of ECAD protein turnover, and PKA is a necessary redox sensor to enable this process. This study provides some mechanistic insight into how oxidative stress-induced ECAD degradation promotes tumor metastasis of HCC.

Development of a Multifaceted Perspective for Systematic Analysis, Assessment, and Performance for Environmental Standards of Contaminated Sites.

Contaminated soil and groundwater can pose significant risks to human health and ecological environments, making the remediation of contaminated sites a pressing and sustained challenge. It is significant to identify key performance indicators and advance environmental management standards of contaminated sites. The traditional study currently focuses on the inflexible collection of related files and displays configurable limitations regarding integrated assessment and in-depth analysis of published standards. In addition, there is a relative lack of research focusing on the analysis of different types of standard documents. Herein, we introduce a cross-systematic retrospective and review for the development of standards of the contaminated sites, including the comprehensive framework, multifaceted analysis, and improved suggestion of soil and groundwater standards related to the environment. The classification and structural characteristics of different types of files are systematically analyzed of over 300 national, trade, local, and group standards for the contaminated sites. It exhibits that trade standards are the main types and testing methods are the important format within numerical considerations of soil standards. The guide standard serves as a crucial component in environmental management for investigating, assessing, and remediating of contaminated sites. Future improvement plans and development directions are proposed for advancing robust technical support for effective soil contamination prevention and control. This multidimensional analysis and the accompanying suggestions can provide improved guidance for Chinese environmental management of contaminated sites and sparkle the application of standards in a wide range of countries.

Deciphering the carbon and nitrogen component conversion in humification process mediated by distinct microbial mechanisms in composting from different domestic organic wastes.

This study aimed to compare the process of maturity and humus fraction evolution as well as bacterial community dynamics in composting from different domestic organic wastes (food waste (FW), and vegetable waste (VW)) and decipher the key biotic influencing factors of humic acid formation through correlation analysis and ecological network. The results showed that organic carbon components in FW with high ratio of soluble organic carbon and hemicellulose were more easily to be degraded in composting compared to VW. After 30 days of composting, the content of HA-C generated by VW was 35.41%, higher than 29.01% of FW, and the growth rate of HA-C generated was 38.42% and 28.34%, respectively. PARAFAC analysis showed that the structure of HA generated in VW was more complex, and the proportion of humic acid-like components (C3 + C4) was 60.32%, while FW only accounted for 43.86%. However, the evolution growth rate of aromatic components in HA in FW was 26.88% in 30 days of compost, which was higher than 15.17% in VW. High-throughput sequencing indicated that Lactobacillus was the initial dominated genera in composting from different domestic wastes. Thermobifida, Thermovum, and Pusillimas as well as Aeribacillus were core bacterial genera that promoted the humification process in FW and VW, respectively. Network analysis showed that there was higher bacterial interacted connection degree and complexity in FW compared to VW. This study was of great significance for optimizing organics conversion and humification efficiency of household waste composting.

Spatiotemporal successions of N, S, C, Fe, and As cycling genes in groundwater of a wetland ecosystem: Enhanced heterogeneity in wet season.

Microorganisms in wetland groundwater play an essential role in driving global biogeochemical cycles. However, largely due to the dynamics of spatiotemporal surface water-groundwater interaction, the spatiotemporal successions of biogeochemical cycling in wetland groundwater remain poorly delineated. Herein, we investigated the seasonal coevolution of hydrogeochemical variables and microbial functional genes involved in nitrogen, carbon, sulfur, iron, and arsenic cycling in groundwater within a typical wetland, located in Poyang Lake Plain, China. During the dry season, the microbial potentials for dissimilatory nitrate reduction to ammonium and ammonification were dominant, whereas the higher potentials for nitrogen fixation, denitrification, methane metabolism, and carbon fixation were identified in the wet season. A likely biogeochemical hotspot was identified in the area located in the low permeable aquifer near the lake, characterized by reducing conditions and elevated levels of Fe2+ (6.65-17.1 mg/L), NH4+ (0.57-3.98 mg/L), total organic carbon (1.02-1.99 mg/L), and functional genes. In contrast to dry season, higher dissimilarities of functional gene distribution were observed in the wet season. Multivariable statistics further indicated that the connection between the functional gene compositions and hydrogeochemical variables becomes less pronounced as the seasons transition from dry to wet. Despite this transition, Fe2+ remained the dominant driving force on gene distribution during both seasons. Gene-based co-occurrence network displayed reduced interconnectivity among coupled C-N-Fe-S cycles from the dry to the wet season, underpinning a less complex and more destabilizing occurrence pattern. The rising groundwater level may have contributed to a reduction in the stability of functional microbial communities, consequently impacting ecological functions. Our findings shed light on microbial-driven seasonal biogeochemical cycling in wetland groundwater.

Inhibiting mtDNA-STING-NLRP3/IL-1ß axis-mediated neutrophil infiltration protects neurons in Alzheimer's disease.

Neutrophil is a pathophysiological character in Alzheimer's disease. The pathogen for neutrophil activation in cerebral tissue is the accumulated amyloid protein. In our present study, neutrophils infiltrate into the cerebra in two models (transgenic model APP/PS1 and stereotactic injection model) and promote neuron apoptosis, releasing their cellular constituents, including mitochondria and mitochondrial DNA (mtDNA). We found that both Aß1-42 and mtDNA could provoke neutrophil infiltration into the cerebra, and they had synergistic effects when they presented together. This neutrophillic neuroinflammation upregulates expressions of STING, NLRP3 and IL-1ß. These inflammatory cytokines with mtDNA constitute the mtDNA-STING-NLRP3/IL-1ß axis, which is the prerequisite for neutrophil infiltration. When any factor in this pathway is depleted, the migration of neutrophils into cerebral tissue is ceased, with neurons and cognitive function being protected. Thus, we provide a novel perspective to alleviate the progression of Alzheimer's disease.

Inoculating functional bacteria improved the humification process by regulating microbial networks and key genera in straw composting by adding different nitrogen sources.

The aim of this study was to compare the effect of functional inoculant and different nitrogen sources on the relationship among lignocellulose, precursors, and humus as well as their interactions with bacterial genera in straw composting. Results showed that inoculation improved the heating process and retained more nitrate compared to control. Inoculation increased the degradation of lignocellulosic components by 26.9%-81.6% and the formation of humus by 15.7%-23.0%. Bioinformatics analysis showed that inoculation enriched key genera Chryseolinea in complex nitrogen source (pig manure) compost and Pusillimas, Luteimonas, and Flavobacteria in single nitrogen source (urea) compost, which were related to humus formation. Network analysis found that inoculation and urea addition improved the microbial synergistic effect and inoculation combined with pig manure had more complex modularity and interactions. Combining the functional bacterial inoculant with urea helped to enhance the degradation of lignocellulose and humification process during straw composting especially with single nitrogen source.

Synergistic effects of chemical additives and mature compost on reducing H2S emission during kitchen waste composting.

Additives could improve composting performance and reduce gaseous emission, but few studies have explored the synergistic of additives on H2S emission and compost maturity. This research aims to make an investigation about the effects of chemical additives and mature compost on H2S emission and compost maturity of kitchen waste composting. The results showed that additives increased the germination index value and H2S emission reduction over 15 days and the treatment with both chemical additives and mature compost achieved highest germination index value and H2S emission reduction (85%). Except for the treatment with only chemical additives, the total sulfur content increased during the kitchen waste composting. The proportion of effective sulfur was higher with the addition of chemical additives, compared with other groups. The relative abundance of H2S-formation bacterial (Desulfovibrio) was reduced and the relative abundance of bacterial (Pseudomonas and Paracoccus), which could convert sulfur-containing substances and H2S to sulfate was improved with additives. In the composting process with both chemical additives and mature compost, the relative abundance of Desulfovibrio was lowest, while the relative abundance of Pseudomonas and Paracoccus was highest. Taken together, the chemical additives and mature compost achieved H2S emission reduction by regulating the dynamics of microbial community.

EpCAM-targeting CAR-T cell immunotherapy is safe and efficacious for epithelial tumors.

The efficacy of CAR-T cells for solid tumors is unsatisfactory. EpCAM is a biomarker of epithelial tumors, but the clinical feasibility of CAR-T therapy targeting EpCAM is lacking. Here, we report pre- and clinical investigations of EpCAM-CAR-T cells for solid tumors. We demonstrated that EpCAM-CAR-T cells costimulated by Dectin-1 exhibited robust antitumor activity without adverse effects in xenograft mouse models and EpCAM-humanized mice. Notably, in clinical trials for epithelial tumors (NCT02915445), 6 (50%) of the 12 enrolled patients experienced self-remitted grade 1/2 toxicities, 1 patient (8.3%) experienced reversible grade 3 leukopenia, and no higher-grade toxicity reported. Efficacy analysis determined two patients as partial response. Three patients showed >23 months of progression-free survival, among whom one patient experienced 2-year progress-free survival with detectable CAR-T cells 200 days after infusion. These data demonstrate the feasibility and tolerability of EpCAM-CAR-T therapy.

Chimeric Antigen Receptor-T Cell Therapy Decreases Distant Metastasis and Inhibits Local Recurrence Post-surgery in Mice.

Distant metastasis and primary tumor relapse are the two main hurdles to the success of surgical treatment for cancer patients. Circulating tumor cells (CTCs) and incomplete surgical resection are the primary cause of distant metastasis and local recurrence of tumors, respectively. Chimeric antigen receptor (CAR)-modified T cells target residual carcinomas and CTCs hold the potential to inhibit primary recurrence and reduce tumor metastasis, but the experimental evidence is lacking. Here, we developed a surgery-induced tumor metastasis model in immunocompetent mice to investigate the efficacy of CAR-T cells therapy in preventing metastasis and local recurrence. We observed that subcutaneous tumor resection has induced a large number of CTCs intravasated into circulation. EpCAM-specific CAR-T was effective in clearing CTCs following surgical removal of the tumor. This resulted in less pulmonary metastasis and longer survival in mice when compared to mice treated with surgery followed by Mock-T cells infusion. In addition, the local relapse was obviously inhibited at the surgical site followed by EpCAM-CAR-T cell treatment. This study demonstrated that CAR-T cell therapy can be an adjuvant treatment following surgery to prevent tumor metastasis and inhibit primary tumor relapse for cancer patients.

Co-inoculation of phosphate-solubilizing bacteria and phosphate accumulating bacteria in phosphorus-enriched composting regulates phosphorus transformation by facilitating polyphosphate formation.

This study aimed to explore the impact of co-inoculating phosphate-solubilizing bacteria (PSB) and phosphate accumulating bacteria (PAB) on phosphorus forms transformation, microbial biomass phosphorus (MBP) and polyphosphate (Poly-P) accumulation, bacterial community composition in composting, using high throughput sequencing, PICRUSt 2, network analysis, structural equation model (SEM) and random forest (RF) analysis. The results demonstrated PSB-PAB co-inoculation (T1) reduced Olsen-P content (1.4 g) but had higher levels of MBP (74.2 mg/kg) and Poly-P (419 A.U.) compared to PSB-only (T0). The mantel test revealed a significantly positive correlation between bacterial diversity and both bioavailable P and MBP. Halocella was identified as a key genus related to Poly-P synthesis by network analysis. SEM and RF analysis showed that pH and bacterial community had the most influence on Poly-P synthesis, and PICRUSt 2 analysis revealed inoculation of PAB increased ppk gene abundance in T1. Thus, PSB-PAB co-inoculation provides a new idea for phosphorus management.

Comparing bacterial dynamics for the conversion of organics and humus components during manure composting from different sources.

The study aimed to compare the differences in organic fractions transformation, humus components and bacterial community dynamics during manure composting from different sources, and to identify the key biotic and abiotic factors driving the humification process. Five types of manure [pig manure (PM), cow dung (CD), sheep manure (SM), chicken manure (CM), and duck manure (DM)] were used as raw materials for 30 days composting. The results showed the obvious difference of organic fractions decomposition with more cellulose degradation in CD and SM composting and more hemicellulose degradation in PM and CM composting. Composting of PM and CD contained significantly higher humus fractions than the other composts. Fluorescence spectra indicated that SM composting tended to form structurally stable humic acid fractions, while CM and DM tended to form structurally complex fulvic acid fractions. Pearson correlation analysis showed that humification process of composts in category A (PM, CD) with higher humification degree than category B (SM, CM, and DM) was positively correlated with lignin and hemicellulose degradation. Bioinformatics analysis found that Lysinibacillus promoted the degradation of hemicellulose and the conversion of fulvic to humic acid in the composts of category A, and in category B, Thermobifida, Lactobacillus, and Ureibacillus were key genera for humic acid formation. Network analysis indicated that bacterial interaction patterns had obvious differences in composting with different humus and humification levels.

Regulation and signaling pathways in cancer stem cells: implications for targeted therapy for cancer.

Cancer stem cells (CSCs), initially identified in leukemia in 1994, constitute a distinct subset of tumor cells characterized by surface markers such as CD133, CD44, and ALDH. Their behavior is regulated through a complex interplay of networks, including transcriptional, post-transcriptional, epigenetic, tumor microenvironment (TME), and epithelial-mesenchymal transition (EMT) factors. Numerous signaling pathways were found to be involved in the regulatory network of CSCs. The maintenance of CSC characteristics plays a pivotal role in driving CSC-associated tumor metastasis and conferring resistance to therapy. Consequently, CSCs have emerged as promising targets in cancer treatment. To date, researchers have developed several anticancer agents tailored to specifically target CSCs, with some of these treatment strategies currently undergoing preclinical or clinical trials. In this review, we outline the origin and biological characteristics of CSCs, explore the regulatory networks governing CSCs, discuss the signaling pathways implicated in these networks, and investigate the influential factors contributing to therapy resistance in CSCs. Finally, we offer insights into preclinical and clinical agents designed to eliminate CSCs.

Secure reversal of immune evasion from refractory NSCLC and highly contagious CoV-2 mutants by using 3D-engineered multifunctional biologics.

There is an imperative choice to develop a secure feasible strategy to address evasion dynamics of refractory tumors and SARS-CoV-2-variants, while stem cell-based protocol may be more reliable as its unique ability for resetting multifunctional immunity to address progressive tumor and the constantly-evolving virus. In this study, spheroid-embryonoid stem cells from mature somatic cells were engineered as multifunctional biologics (3D-E/BSC) and inoculated in senile rhesus to identify secure potential against immune-evasion from viral-variants. Meanwhile, a cohort of eligible patients with stage IV NSCLC were approved for phase I clinical trials. Subsequently, long-lasting security and efficacy were validated by primate and clinical trials (p < 0.01) in that it could not only stimulate serological immunity, but also reset core immunity for hosts to address variant evasion after 3D-E/BSC withdrawal. Particularly, illustrated by single-cell evolving trajectory, 3D-E/BSC had securely reset senile thymus of aging hosts to remodel core immunity by rearranging naive rhythm to evolve TRGC2+/JCHAIN+NKT clusters to abolish tumoral and viral evasion dynamics with path-feedbacks of NSCLC and COVID-19 simultaneously activated, leading to continuous blockade of breakthrough infection of viral-mutants and long-term survival in one-third of terminal patients without adjuvant required. Our study may pioneer a practical multifunctional strategy to eliminate evasion of SARS-CoV-2 variants and refractory NSCLC so as for victims to restart a new life-equation.

Cryo-EM structures of human GPR34 enable the identification of selective antagonists.

GPR34 is a functional G-protein-coupled receptor of Lysophosphatidylserine (LysoPS), and has pathogenic roles in numerous diseases, yet remains poorly targeted. We herein report a cryo-electron microscopy (cryo-EM) structure of GPR34 bound with LysoPS (18:1) and Gi protein, revealing a unique ligand recognition mode with the negatively charged head group of LysoPS occupying a polar cavity formed by TM3, 6 and 7, and the hydrophobic tail of LysoPS residing in a lateral open hydrophobic groove formed by TM3-5. Virtual screening and subsequent structural optimization led to the identification of a highly potent and selective antagonist (YL-365). Design of fusion proteins allowed successful determination of the challenging cryo-EM structure of the inactive GPR34 complexed with YL-365, which revealed the competitive binding of YL-365 in a portion of the orthosteric binding pocket of GPR34 and the antagonist-binding-induced allostery in the receptor, implicating the inhibition mechanism of YL-365. Moreover, YL-365 displayed excellent activity in a neuropathic pain model without obvious toxicity. Collectively, this study offers mechanistic insights into the endogenous agonist recognition and antagonist inhibition of GPR34, and provides proof of concept that targeting GPR34 represents a promising strategy for disease treatment.