Massive Accumulation Research Articles

Replication-Poison Treatment in BRCA1-Deficient Breast Cancer Causes MRE11 Over-Resection that Induces Single-Stranded DNA Accumulation and Mitotic Catastrophe.

BRCA1 deficiency is observed in approximately 25% of triple-negative breast cancer (TNBC). BRCA1, a key player of homologous recombination (HR) repair, is also involved in stalled DNA replication fork protection and repair. Here, we investigated the sensitivity of BRCA1-deficient TNBC models to the frequently used replication chain terminator gemcitabine, which does not directly induce DNA breaks. A large fraction of BRCA1-deficient cells was sensitive to gemcitabine, in contrast to their isogenic BRCA1-proficient counterparts. Gemcitabine treated BRCA1-deficient cells accumulated massive levels of single strand DNA (ssDNA) and presented no RPA or RAD51 nuclear foci. The gemcitabine-induced accumulation of ssDNA in BRCA1-deficient cells was strongly diminished by targeting MRE11 with inhibitors and by siRNA attenuation. In contrast, treatment with the PARP1/2 inhibitor olaparib did not result in MRE11 dependent over-resection. Furthermore, a fraction of gemcitabine treated BRCA1-deficient cells that showed massive ssDNA accumulation slipped into mitosis, producing mitotic bridges and strongly stained BrdU and γH2AX micronuclei (MN). The BrdU-positive MN and DNA bridges also stained positively for cGAS. In conclusion, these data suggest that gemcitabine treatment in BRCA1-deficient TNBC exposes unprotected nascent DNA linked to replication fork reversal, which leads to MRE11 over-resection and ssDNA accumulation. Therefore, the observed hypersensitivity to gemcitabine indicates that it could be a beneficial addition to BRCA1-deficient TNBC treatment.

Sudden Death of a Four-Day-Old Newborn Due to Mitochondrial Trifunctional Protein/Long-Chain 3-Hydroxyacyl-CoA Dehydrogenase Deficiencies and a Systematic Literature Review of Early Deaths of Neonates with Fatty Acid Oxidation Disorders

Mitochondrial trifunctional protein (MTP) and long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiencies have been a part of the Slovenian newborn screening (NBS) program since 2018. We describe a case of early lethal presentation of MTPD/LCHADD in a term newborn. The girl was born after an uneventful pregnancy and delivery, and she was discharged home at the age of 3 days, appearing well. At the age of 4 days, she was found without signs of life. Resuscitation was not successful. The NBS test performed using tandem mass spectrometry (MS/MS) showed a positive screen for MTPD/LCHADD. Genetic analysis performed on a dried blood spot (DBS) sample identified two heterozygous variants in the HADHA gene: a nucleotide duplication introducing a premature termination codon (p.Arg205Ter) and a nucleotide substitution (p.Glu510Gln). Post-mortem studies showed massive macro-vesicular fat accumulation in the liver and, to a smaller extent, in the heart, consistent with MTPD/LCHADD. A neonatal acute cardiac presentation resulting in demise was suspected. We conducted a systematic literature review of early neonatal deaths within 14 days postpartum attributed to confirmed fatty acid oxidation disorders (FAODs), which are estimated to account for 5% of sudden infant deaths. We discuss the pitfalls of the NBS for MTPD/LCHADD.

Lactate accumulation promotes immunosuppression and fibrotic transformation of bone marrow microenvironment in myelofibrosis

BackgroundClonal myeloproliferation and fibrotic transformation of the bone marrow (BM) are the pathogenetic events most commonly occurring in myelofibrosis (MF). There is great evidence indicating that tumor microenvironment is characterized by high lactate levels, acting not only as an energetic source, but also as a signaling molecule.MethodsTo test the involvement of lactate in MF milieu transformation, we measured its levels in MF patients’ sera, eventually finding a massive accumulation of this metabolite, which we showed to promote the expansion of immunosuppressive subsets. Therefore, to assess the significance of its trafficking, we inhibited monocarboxylate transporter 1 (MCT1) by its selective antagonist, AZD3965, eventually finding a mitigation of lactate-mediated immunosuppressive subsets expansion. To further dig into the impact of lactate in tumor microenvironment, we evaluated the effect of this metabolite on mesenchymal stromal cells (MSCs) reprogramming.ResultsOur results show an activation of a cancer-associated phenotype (CAF) related to mineralized matrix formation and early fibrosis development. Strikingly, MF serum, enriched in lactate, causes a strong deposition of collagen in healthy stromal cells, which was restrained by AZD3965. To corroborate these outcomes, we therefore generated for the first time a TPOhigh zebrafish model for the establishment of experimental fibrosis. By adopting this model, we were able to unveil a remarkable increase in lactate concentration and monocarboxylate transporter 1 (MCT1) expression in the site of hematopoiesis, associated with a strong downregulation of lactate export channel MCT4. Notably, exploiting MCTs expression in biopsy specimens from patients with myeloproliferative neoplasms, we found a loss of MCT4 expression in PMF, corroborating changes in MCT expression during BM fibrosis establishment.ConclusionsIn conclusion, our results unveil lactate as a key regulator of immune escape and BM fibrotic transformation in MF patients, suggesting MCT1 blocking as a novel antifibrotic strategy.

Casein Kinases 2-dependent phosphorylation of the placental ligand VAR2CSA regulates Plasmodium falciparum-infected erythrocytes cytoadhesion.

Placental malaria is characterized by the massive accumulation and sequestration of infected erythrocytes in the placental intervillous blood spaces, causing severe birth outcomes. The variant surface antigen VAR2CSA is associated with Plasmodium falciparum sequestration in the placenta via its capacity to adhere to chondroitin sulfate A. We have previously shown that the extracellular region of VAR2CSA is phosphorylated on several residues and that the phosphorylation enhances the adhesive properties of CSA-binding infected erythrocytes. Here, we aimed to identify the kinases mediating this phosphorylation. We report that human and Plasmodium falciparum Casein Kinase 2α are involved in the phosphorylation of the extracellular region of VAR2CSA. We notably show that both CK2α can phosphorylate the extracellular region of recombinant and immunoprecipitated VAR2CSA. Mass spectrometry analysis of recombinant VAR2CSA phosphorylated by recombinant Human and P. falciparum CK2α combined with site-directed mutagenesis led to the identification of residue S1068 in VAR2CSA, which is phosphorylated by both enzymes and is associated with CSA binding. Furthermore, using CRISPR/Cas9 we generated a parasite line in which phosphoresidue S1068 was changed to alanine. This mutation strongly impairs infected erythrocytes adhesion by abolishing VAR2CSA translocation to the surface of infected erythrocytes. We also report that two specific CK2 inhibitors reduce infected erythrocytes adhesion to CSA and decrease the phosphorylation of the recombinant extracellular region of VAR2CSA using either infected erythrocytes lysates as a source of kinases or recombinant Human and P. falciparum casein kinase 2. Taken together, these results undoubtedly demonstrate that host and P. falciparum CK2α phosphorylate the extracellular region of VAR2CSA and that this post-translational modification is important for VAR2CSA trafficking and for infected erythrocytes adhesion to CSA.

Impact of inner hydrophobicity of dendrimer nanomicelles on biodistribution: a PET imaging study.

Self-assembly is a powerful strategy for building nanosystems for biomedical applications. We have recently developed small amphiphilic dendrimers capable of self-assembling into nanomicelles for tumor imaging. In this context, we studied the impact of increased hydrophobicity of the amphiphilic dendrimer on hydrophilic/hydrophobic balance and consequently on the self-assembly and subsequent biodistribution. Remarkably, despite maintaining the exact same surface chemistry, similar zeta potential, and small size, the altered and enlarged hydrophobic component within the amphiphilic dendrimer led to enhanced stability of the self-assembled nanomicelles, with prolonged circulation time and massive accumulation in the liver. This study reveals that even structural alteration within the interior of nanomicelles can dramatically impact biodistribution profiles. This finding highlights the deeper complexity of rational design for nanomedicine and the need to consider factors other than surface charge and chemistry, as well as size, all of which significantly impact the biodistribution of self-assembling nanosystems.

Tagless LysoIP for immunoaffinity enrichment of native lysosomes from clinical samples.

Lysosomes are implicated in a wide spectrum of human diseases including monogenic lysosomal storage disorders (LSDs), age-associated neurodegeneration and cancer. Profiling lysosomal content using tag-based lysosomal immunoprecipitation (LysoTagIP) in cell and animal models has substantially moved the field forward, but studying lysosomal dysfunction in human patients remains challenging. Here, we report the development of the 'tagless LysoIP' method, designed to enable the rapid enrichment of lysosomes, via immunoprecipitation, using the endogenous integral lysosomal membrane protein TMEM192, directly from clinical samples and human cell lines (e.g., induced pluripotent stem cell derived neurons). Isolated lysosomes were intact and suitable for subsequent multimodal omics analyses. To validate our approach, we applied the tagless LysoIP to enrich lysosomes from peripheral blood mononuclear cells derived from fresh blood of healthy donors and patients with CLN3 disease, an autosomal recessive neurodegenerative LSD. Metabolic profiling of isolated lysosomes revealed massive accumulation of glycerophosphodiesters (GPDs) in patients' lysosomes. Interestingly, a patient with a milder phenotype and genotype displayed lower accumulation of lysosomal GPDs, consistent with their potential role as disease biomarkers. Altogether, the tagless LysoIP provides a framework to study native lysosomes from patient samples, identify disease biomarkers, and discover human-relevant disease mechanisms.

Harnessing ROS Amplification and GSH Depletion Using a Carrier-Free Nanodrug to Enhance Ferroptosis-Based Cancer Therapy.

Ferroptosis, a non-apoptotic form of cell death characterized by the production of reactive oxygen species (ROS) and massive accumulation of lipid peroxidation (LPO), shows significant promise in cancer therapy. However, the overexpression of glutathione (GSH) at the tumor site and insufficient ROS often result in unsatisfactory therapeutic efficacy. A multistage, GSH-consuming, and ROS-providing carrier-free nanodrug capable of efficiently loading copper ions (Cu2+), sorafenib (SRF), and chlorogenic acid (CGA) (Cu2+-CGA-SRF, CCS-NDs) is developed to mediate enhanced ferroptosis therapy. Through a reductive intracellular environment, Cu2+ in the CCS-NDs reacted with intracellular GSH, alleviating the antioxidant capacity of tumor tissues and triggering the release of drugs. Meanwhile, the released SRF inhibited system xc-, thereby blocking cystine uptake and reducing GSH synthesis in tumor cells. By depleting stored GSH and inhibiting its synthesis, CCS-NDs achieved efficient GSH depletion and increased accumulation of toxic LPO. More importantly, the high concentration of CGA in the CCS-NDs induced ROS generation, further promoting ferroptosis. Both in vitro and in vivo results demonstrated that CCS-NDs effectively triggered ferroptosis in tumor cells by inactivating glutathione peroxidase 4 and inducing LPO. Overall, the carrier-free nanodrug CCS-NDs offer a promising strategy for regulating GSH and LPO levels in ferroptosis-based cancer therapy.

HMOX1-LDHB interaction promotes ferroptosis by inducing mitochondrial dysfunction in foamy macrophages during advanced atherosclerosis.

Advanced atherosclerosis is the pathological basis for acute cardiovascular events, with significant residual risk of recurrent clinical events despite contemporary treatment. The death of foamy macrophages is a main contributor to plaque progression, but the underlying mechanisms remain unclear. Bulk and single-cell RNA sequencing demonstrated that massive iron accumulation in advanced atherosclerosis promoted foamy macrophage ferroptosis, particularly in low expression of triggering receptor expressed on myeloid cells 2 (TREM2low) foamy macrophages. This cluster exhibits metabolic characteristics with low oxidative phosphorylation (OXPHOS), increasing ferroptosis sensitivity. Mechanically, upregulated heme oxygenase 1 (HMOX1)-lactate dehydrogenase B (LDHB) interaction enables Lon peptidase 1 (LONP1) to degrade mitochondrial transcription factor A (TFAM), leading to mitochondrial dysfunction and ferroptosis. Administration of the mitochondria-targeted reactive oxygen species (ROS) scavenger MitoTEMPO (mitochondrial-targeted TEMPO) or LONP1 inhibitor bortezomib restored mitochondrial homeostasis in foamy macrophages and alleviated atherosclerosis. Collectively, our study elucidates the cellular and molecular mechanism of foamy macrophage ferroptosis, offering potential therapeutic strategies for advanced atherosclerosis.

In Situ Transformable Nanoparticle Effectively Suppresses Bladder Cancer by Damaging Mitochondria and Blocking Mitochondrial Autophagy Flux.

Tumor therapeutic strategies based on mitochondrial damage have become an emerging trend. However, the low drug delivery efficiency caused by lysosomal sequestration and the activation of protective mitochondrial autophagy severely restricts the therapeutic efficacy. Herein, an in situ transformable nanoparticle named KCKT is developed to promote lysosomal escape and directly damage mitochondria while blocking mitochondrial autophagy. KCKT exhibits acid responsiveness for precise self-assembly into nanofibers within the lysosomes of cancer cells. The massive accumulation of nanofibers and excessive production of reactive oxygen species (ROS) under sonodynamic therapy synergistically induce lysosomal damage. This facilitates the escape of nanofibers from lysosomal sequestration, thereby enhancing drug delivery. Subsequently, the escaped nanofibers specifically aggregate around the mitochondria for long-term retention and generate ROS under ultrasound irradiation to induce mitochondrial damage. Notably, due to lysosomal dysfunction, damaged mitochondria cannot be cleared by autophagy, further aggravating oxidative damage. These results reveal that KCKT effectively improves drug delivery and mitochondria-targeted therapy efficiency by blocking protective autophagy. These findings hold significant potential for advancing the field of mitochondria-targeted therapy.

Shape and Size Distribution of Sulfide Globules in the Raglan Magmatic Sulfide Deposit: Implications for Deposition and Exploration of Massive Sulfide Orebodies

Abstract Globular sulfide is the best-preserved textural representation of immiscible sulfide liquids in silicate magmas, containing valuable information about the mechanisms of their transport and deposition and the formation of magmatic sulfide deposits. Previous studies have indicated that sulfide globule textures may convey useful information about their proximity to massive sulfide accumulations. This study quantitatively evaluates the genetic and spatial relationships between globular sulfides and massive orebodies in Zones 8 and 14 of the Raglan Horizon of northern Quebec by investigating their geochemical characteristics and systematically measuring globule size distributions (GSDs) across this portion of the Raglan deposit group. Their compositions suggest that sulfide globules, disseminated sulfides, net-textured sulfides, and massive sulfides in Zones 8 and 14 are genetically related and geochemically indistinguishable. The sulfide GSDs show that most samples taken from locations distal to massive lenses exhibit a simple log-linear relationship resembling the result of a single homogeneous nucleation event with linear growth and relatively constant nucleation density, or disaggregation of sulfide droplets by ligament stretching. In contrast, most samples of the proximal population show kinked GSD shapes. The kinked profiles can be attributed to processes possibly including mechanical sorting, Ostwald ripening, coalescence of sulfide globules, or the mixing of two globule populations with different size distributions. Whereas mechanical sorting, Ostwald ripening, and coalescence between sulfide globules are considered as, at most, minor contributors to the GSD shapes observed in this region, the kinked GSDs are best explained as representations of a mixed population of large, transported globules deposited in an early stage and finer-grained sulfide droplets deposited in a latter stage. Based on these interpretations, an ore-forming mechanism is proposed which starts with (1) deposition of large sulfide globules in a footwall embayment during turbulent magma flow in which the finer globules remain in suspension, followed by (2) the deposition and entrapment of fine-grained sulfide microdroplets due to a transition from turbulent to a transitional or laminar flow regime, and eventually (3) the rapid downward percolation of sulfide microdroplets through the pore network of the cumulate pile to form massive sulfide pools on the hard substrate of the footwall embayment. Furthermore, we find a close spatial relationship between samples with kinked GSDs and high globule number densities and massive sulfide ores. We demonstrate the robustness of using globule number density as a pointer to massive sulfide accumulations by successfully predicting the actual locations of orebodies 8M, 14K, and 14J in a 3-D space. Overall, we suggest that the texture of large globules immersed in finely dispersed clouds of abundant disseminated small sulfide globules is a strong indicator for proximity to massive sulfide accumulations, and the recognition of this texture may provide a critical tool for future exploration for massive magmatic sulfide ore lenses. We present schematic illustrations of favorable and unfavorable GSDs to aid in the qualitative application of these concepts during logging of exploration drill core.

Feline cumulus cells and oocytes show massive accumulation of lipid droplets and upregulation of PLIN2 expression after in vitro maturation

This study investigates lipid content and expression of genes involved in lipid metabolism in feline cumulus cells and oocytes before and after in vitro maturation (IVM). Domestic cats represent valuable models in reproductive research, yet the efficiency of in vitro embryo production remains suboptimal, with contributing factors still under investigation. We characterized lipid droplets (LDs) in oocytes collected from adult queens, both before and after IVM, using confocal microscopy with Bodipy 493/503 staining. We also quantified the expression of four genes involved in regulation of lipid metabolism. Our findings revealed a substantial accumulation of LDs and a significant upregulation of PLIN2 expression in both cumulus cells and oocytes following IVM. The number and total area of LDs and fluorescence intensity increased markedly in oocytes at the MII stage, while the average LD diameter decreased. Similarly, cumulus cells showed an increase in number and total area of LDs post-IVM, suggesting their involvement in oocyte maturation by modulating lipid homeostasis. This study presents the first detailed characterization of lipid droplet dynamics in feline oocytes and cumulus cells, providing insights into metabolic processes during IVM, which are critical for optimizing assisted reproductive technologies in felids.

Collagenofibrotic Glomerulopathy with Thrombotic Microangiopathy: A Rare Case Report

Collagenofibrotic Glomerulopathy (CG) is a very rare glomerular disease characterised by the massive accumulation of atypical type III collagen fibrils within the mesangial matrix and subendothelial space, as well as a marked increase in serum type III procollagen peptide levels. Owing to its rarity, the cause and pathogenesis of CG remain uncertain. While initial reports of CG were primarily in patients of Asian descent, subsequent cases in patients of European descent have also been published. Clinically, patients present with oedema and hypertension, often progressing to end-stage renal disease. Proteinuria is a cardinal manifestation of this disease. A definitive diagnosis is contingent upon recognising this entity, maintaining a high index of suspicion, and confirming it through immunohistochemistry and Electron Microscopy (EM), due to its rarity, non specific presenting signs and symptoms, and various differential diagnoses on histology. Here, a case of a 44-year-old non diabetic, hypertensive female who presented with pedal oedema and nephrotic range proteinuria. The biopsy showed nodular expansion of the mesangium by homogeneous, congo red-negative, eosinophilic material, along with vascular changes indicative of acute thrombotic microangiopathy. Many conditions, such as amyloidosis, fibrillary glomerulonephritis, diabetic nephropathy, fibronectin glomerulopathy, light chain deposition disease, and thrombotic microangiopathy, can cause nodular or membranoproliferative glomerular changes. The biopsy was initially reported as nodular glomerulosclerosis, which, upon EM, showed mesangial and subendothelial space expansion by banded fibrillary material. Further immunohistochemistry for type III collagen was performed, which yielded positive results. Thus, the diagnosis of CG was confirmed.

Coal gangue based geopolymer as an additive of warm mix asphalt: preparation and properties

Abstract In contrast to conventional hot mix asphalt technology (HMA), warm mix asphalt technology (WMA) permits the mixing and compaction of asphalt mixture at lower temperatures. This innovative approach is characterized by significant energy savings and reduced emissions, making it a sustainable choice for asphalt mixture production. This paper used coal gangue to prepare a WMA additive. By leveraging the free water in its zeolite-like structure, which evaporates upon heating, bubbles form within the bitumen. This causes bitumen to expand and lose viscosity, thereby achieving the purpose of warm mixing. Both achieve a decline in the asphalt mixture’s mixing temperature and provide an effective method for utilizing coal gangue resources. It helps mitigate the environmental hazards associated with the massive accumulation of coal gangue, including soil contamination and water resource pollution. Therefore, based on alkali activation theory, this paper aimed to develop a novel coal gangue geopolymer (CG) WMA additive. CG was characterized through microscopic testing and analysis of water storage stability. The impact of varying CG dosages on the conventional performance of bitumen was examined. The bitumen foaming test elucidated the warm mixing mechanism and foaming behavior of the CG. Additionally, the road performance of the WMA was analyzed. The findings indicate that CG has abundant pore structures, providing an effective water storage space, and CG as a WMA additive exhibits good water storage stability at ambient temperature. When the CG dosage is 5% by the mass of bitumen, the viscosity of bitumen at 130∼140 °C is appropriate, and the WMA results in good road performance. Compared with HMA, the mixing temperature is reduced by a range of 20∼50 °C.

Study on the Effect of Basalt Fiber Content and Length on Mechanical Properties and Durability of Coal Gangue Concrete

The massive accumulation of coal gangue not only causes a waste of resources but also brings serious environmental pollution problems. To promote the utilization of coal gangue resources, mitigate environmental pollution from coal gangue, and address the shortage of natural aggregates, this study investigates the use of coal gangue to replace coarse aggregate at a 40% replacement rate to prepare coal gangue concrete (CGC). The current research on the modification of gangue concrete by BF has been less often compared with the research on the effect of basalt fiber (BF) on the properties of ordinary concrete, so in this study, BF with different admixtures and lengths were added into CGC. Additionally, basalt fibers (BFs) of varying amounts and lengths were incorporated into CGC. The study explored the effects of BF on the tensile strength, splitting tensile strength, and flexural strength of CGC. It was found that the mechanical properties of CGC improved significantly when the BF dosage was 0.10–0.15% and the length was 18 mm. This is evidenced by an increase in the compressive strength of 3.94–5.11%, split tensile strength of 11.20–16.18%, and flexural strength of 8.23–12.97%. BF was able to refine pore space, prevent crack development, and bridge cracks in CGC. To further investigate the effect of BF on the long-term service performance of CGC, the effects of BF on the appearance, quality, and compressive strength of CGC in sulfate and freeze–thaw environments were examined. The results indicated that a BF dosage of 0.10–0.15% significantly enhanced the sulfate erosion resistance and freeze–thaw resistance of CGC. This is shown by a 36.76–46.90% reduction in the rate of loss of compressive strength of CGC under the freeze–thaw cycling and a 6.21–8.50% increase in the corrosion resistance factor of CGC under a sulfate attack. BF improved the pore structure and reduced seepage channels, thereby enhancing the durability of CGC.

Physiological response of microalga Dunaliella parva when treated with MeJA, GA3.

DpAP2 is a transcription factor regulating carotenoid biosynthesis pathway. It was speculated that MeJA significantly decreased expression of DpAP2 gene, then the decreasing DpAP2 expression significantly inhibited expression of some key enzyme genes such as PSY, PDS and GGPS in carotenoid biosynthesis pathway. In contrast, it was speculated that GA3 significantly increased expression of DpAP2 gene, then the increasing DpAP2 expression significantly increased expression of some key enzyme genes such as PDS and GGPS in carotenoid biosynthesis pathway. To increase the content of carotenoid, we evaluated the effect of DpAP2 overexpression on carotenoid accumulation in D. parva. Transgenic D. parva showed a higher carotenoid content (3.18 mg/g DW) compared with control group (2.13 mg/g DW) at 9 d. The dosage effects of exogenous hormones MeJA and GA3 were found in D. parva cells treated with different concentrations of MeJA (10, 20, 50, 100 μM) and GA3 (10, 20, 50, 100 μM). The high concentrations of MeJA (10-100 μM) inhibited the accumulation of carotenoid, and the relative expression of DpAP2, PSY, PDS and GGPS decreased significantly. On the contrary, the relative expression of DpAP2, PDS and GGPS increased significantly when D. parva was treated with 10, 20, 50 and 100 μM GA3, which promoted the biosynthesis of carotenoid. Therefore, we inferred that there was a hierarchical regulation from hormone, transcription factor, key enzyme gene to carotenoid accumulation in carotenoid biosynthesis. Carotenoid biosynthesis was enhanced by DpAP2 overexpression (1.4930 fold of control) and exogenous substances such as GA3 (1.5889 fold of control), which laid a foundation for massive accumulation of carotenoids in microalgae. In the future, further studies were required to demonstrate the complex regulatory network.

The Nr4a family regulates intrahepatic Treg proliferation and liver fibrosis in MASLD models.

Nonalcoholic steatohepatitis (NASH) is a chronic progressive liver disease and highly prevalent worldwide. NASH is characterized by hepatic steatosis, inflammation, fibrosis and liver damage, which eventually results in liver dysfunction due to cirrhosis or hepatocellular carcinoma. However, the cellular and molecular mechanisms underlying NASH progression remain largely unknown. Here, we found an increase of Nr4a family of orphan nuclear receptors expression in intrahepatic T cells from mice with diet-induced NASH. Loss of Nr4a1 and Nr4a2 in T cell (dKO) ameliorated liver cell death and fibrosis, thereby mitigating liver dysfunction in NASH mice. dKO resulted in reduction of infiltrated macrophages and Th1/Th17 cells, whereas massive accumulation of T regulatory (Treg) cells in the liver of NASH mice. Combined single-cell RNA transcriptomic and TCR sequencing analysis revealed that intrahepatic dKO Tregs exhibited enhanced TIGIT and IL10 expression and were clonally expanded during NASH progression. Mechanistically, we found that dKO Tregs expressed high levels of Batf which promotes Treg cell proliferation and function upon TCR stimulation. Collectively, our findings not only provide an insight into the impact of intrahepatic Treg cells on NASH pathogenesis, but also suggest a therapeutic potential of targeting of Nr4a family to treat the disease.

Diethyl aminoethyl hexanoate ameliorates salt tolerance associated with ion transport, osmotic adjustment, and metabolite reprograming in white clover

BackgroundSoil salinization is a serious environmental hazard, limiting plant growth and production in different agro-ecological zones worldwide. Diethyl aminoethyl hexanoate (DA-6) as an essential plant growth regulator (PGR) exhibits a beneficial role in improving crop growth and stress tolerance. However, the DA-6-regulated effect and mechanism of salt tolerance in plants are still not fully understood. The objective of current study was to disclose salt tolerance induced by DA-6 in relation to changes in water and redox balance, photosynthetic function, ionic homeostasis, and organic metabolites reprogramming in white clover (Trifolium repens).ResultsA prolonged duration of salt stress caused water loss, impaired photosynthetic function, and oxidative injury to plants. However, foliar application of DA-6 significantly improved osmotic adjustment (OA), photochemical efficiency, and cell membrane stability under salt stress. In addition, high salinity induced massive accumulation of sodium (Na), but decreased accumulation of potassium (K) in leaves and roots of all plants. DA-6-treated plants demonstrated significantly higher transcript levels of genes involved in uptake and transport of Na and K such as VP1, HKT8, SOS1, NHX2, NHX6, and SKOR in leaves as well as VP1, HKT1, HKT8, H+-ATPase, TPK5, SOS1, NHX2, and SKOR in roots. Metabolomics analysis further illustrated that DA-6 primarily induced the accumulation of glucuronic acid, hexanoic acid, linolenic acid, arachidonic acid, inosose, erythrulose, galactopyranose, talopyranose, urea, 1-monopalmitin, glycerol monostearate, campesterol, stigmasterol, and alanine.ConclusionsThe DA-6 significantly up-regulated transcript levels of multiple genes associated with increased Na+ compartmentalization in vacuoles and Na+ sequestration in roots to reduce Na+ transport to photosynthetic organs, thereby maintaining Na+ homeostasis under salt stress. The accumulation of many organic metabolites induced by the DA-6 could be attributed to enhanced cell wall and membrane structural stability and functionality, OA, antioxidant defense, and downstream signal transduction in leaves under salt stress. The present study provides a deep insight about the synergistic role of DA-6 in salt tolerance of white clover.

The fatty acid receptor CD36 promotes macrophage infiltration via p110γ signaling to stimulate metastasis

IntroductionMetabolic regulators are key in controlling immune cell fate in the tumor microenvironment. The accumulation of tumor-associated macrophages (TAMs) in cancer greatly contributes to metastasis and poor outcome. However, the metabolic pathways responsible for TAM accumulation are largely unknown. ObjectiveThis study aims to elucidate the role of the fatty acid translocase CD36 in the regulation of TAM accumulation. MethodsThe immune profile was analyzed in patients with liver metastasis by CIBERSORT. Immunohistostaining of CD68 and CD36 was conducted in clinical specimens from patients with liver metastasis. Myeloid-specific CD36 knockout mice and their littermates were used to establish preclinical liver metastasis models. Subsequently, a series of experiments were used to explore the underlying mechanisms of how CD36 regulates TAM population. ResultsWe found that massive TAM accumulation in patients with liver metastasis is associated with an upregulation of CD36 on TAMs. Liver metastasis is abundantly infiltrated by TAMs that are derived from circulating monocytes, but not tissue-resident macrophages. Myeloid-specific CD36 knockout specifically reduced and inactivated monocyte-differentiated macrophages, resulting in diminished immune suppression and attenuated liver metastasis. The protect effects of CD36 knockout can be abrogated by blockade of macrophage recruitment through CCR2 or the p110γ isoform of PI3K downstream of it. Mechanically, CD36 reprogrammed the lipid metabolism of macrophages, in which sphingolipids were significantly downregulated, that contributed to weakened lipid raft-dependent activation of p110γ. ConclusionCD36 expands TAM population by promoting the recruitment of circulating monocytes through CCL2/CCR2/p110γ signaling. Our findings provide evidence for targeting CD36 as a therapeutic strategy against liver metastasis.

Developing Big Data anomaly dynamic and static detection algorithms: AnomalyDSD spark package

BackgroundAnomaly detection is the process of identifying observations that differ greatly from the majority of data. Unsupervised anomaly detection aims to find outliers in data that is not labeled, therefore, the anomalous instances are unknown. The exponential data generation has led to the era of Big Data. This scenario brings new challenges to classic anomaly detection problems due to the massive and unsupervised accumulation of data. Traditional methods are not able to cop up with computing and time requirements of Big Data problems. MethodsIn this paper, we propose four distributed algorithm designs for Big Data anomaly detection problems: HBOS_BD, LODA_BD, LSCP_BD, and XGBOD_BD. They have been designed following the MapReduce distributed methodology in order to be capable of handling Big Data problems. ResultsThese algorithms have been integrated into an Spark Package, focused on static and dynamic Big Data anomaly detection tasks, namely AnomalyDSD. Experiments using a real-world case of study have shown the performance and validity of the proposals for Big Data problems. ConclusionsWith this proposal, we have enabled the practitioner to efficiently and effectively detect anomalies in Big Data datasets, where the early detection of an anomaly can lead to a proper and timely decision.

Effects of different loads of Pinus pinaster Ait. litter and ash on the germination and growth of Cistus ladanifer L

Key messageCistus ladanifer L. shows an extraordinary plasticity of germination and growth. Fire promotes the regeneration of C. ladanifer by triggering its germination through thermal shock as well as by enhancing its seedling growth through the addition of ashes. On the contrary, stacking of Pinus pinaster Ait. leaf litter may hinder seedling establishment, at least in the first months after germination.ContextPinus pinaster Ait. is affected by forest fires in the Mediterranean basin. Its extreme flammability is due to the high spatial continuity together with the massive accumulation of leaf litter. Cistus ladanifer L. is a species which is widely distributed in the west Mediterranean region where it can form vast shrublands and flourish under these pinewoods. Although high temperatures associated with the occurrence of fire trigger germination of C. ladanifer, knowledge on how other factors, such as the presence of litter on the forest floor or the ash left after the fire, influence germination and seedling growth is essential to improve land management plans.AimsThe main objective of this study was to determine the effects of different loads of litter from P. pinaster and ash from their combustion on the germination and growth of seedlings of C. ladanifer.MethodsTwo experiments were carried out to assess the effect of heat shock (100 °C for 10 min) and two loads of litter and ash from P. pinaster (control, high and low loads of litter, high and low loads of ash) on the germination and growth after 2 and 4 months of C. ladanifer.ResultsHeat shock significantly increased the germination of C. ladanifer. In contrast, the addition of litter and ash had no effect on total germination but affected the growth of the seedlings coming from seeds exposed to heat shock. Litter treatments reduced biomass of 2 months seedlings and ash increased biomass of 4 months seedlings.ConclusionC. ladanifer is a species favoured by fire in different ways depending on the stage of regeneration. Germination is promoted by heat shock while seedling growth is favoured by ash nutrients. In contrast, pine leaf litter hinders seedling growth, although this effect disappears 4 months after germination. Alternative forest management practices to prescribed fires are recommended if preventing the spread of Cistus is a priority.