Potential of amino acids-modified biochar in mitigating the soil Cu and Ni stresses - Targeting the tomato growth, physiology and fruit quality.

Trace heavy metals (HMs) such as copper (Cu) and nickel (Ni) are toxic to plants, especially tomato at high levels. In this study, biochar (BC) was treated with amino acids (AA) to enhance amino functional groups, which effectively alleviated the adverse effects of heavy metals (HMs) on tomato growth. Hence, this study aimed to evaluate the effect of glycine and alanine modified BC (GBC/ABC) on various tomato growth parameters, its physiology, fruit yield and Cu/Ni uptake under Cu and Ni stresses. In a pot experiment, there was 21 treatments with three replications having two rates of simple BC and glycine/alanine enriched BC (0.5% and 1% (w/w). Cu and Ni stresses were added at 150 mg kg-1 respectively. Plants were harvested after 120 days of sowing and subjected to various analysis. Under Cu and Ni stresses, tomato roots accumulated more Cu and Ni than shoots and fruits, while GBC and ABC application significantly enhanced the root and shoot dry weight irrelevant to the stress conditions. Both rates of GBC decreased the malondialdehyde and hydrogen peroxide levels in plants. The addition of 0.5% GBC with Cu enhanced the tomato fruit dry weight by 1.3 folds in comparison to the control treatment; while tomato fruit juice content also increased (50%) in the presence of 0.5% GBC with Ni as compared to control. In summary, these results demonstrated that lower rate of GBC∼0.5% proved to be the best in mitigating the Cu and Ni stress on tomato plant growth by enhancing the fruit production.

Necroptosis contributes to non-alcoholic fatty liver disease pathoetiology with promising diagnostic and therapeutic functions.

Nonalcoholic fatty liver disease (NAFLD) is the most prevalent type of chronic liver disease. However, the disease is underappreciated as a remarkable chronic disorder as there are rare managing strategies. Several studies have focused on determining NAFLD-caused hepatocyte death to elucidate the disease pathoetiology and suggest functional therapeutic and diagnostic options. Pyroptosis, ferroptosis, and necroptosis are the main subtypes of non-apoptotic regulated cell deaths (RCDs), each of which represents particular characteristics. Considering the complexity of the findings, the present study aimed to review these types of RCDs and their contribution to NAFLD progression, and subsequently discuss in detail the role of necroptosis in the pathoetiology, diagnosis, and treatment of the disease. The study revealed that necroptosis is involved in the occurrence of NAFLD and its progression towards steatohepatitis and cancer, hence it has potential in diagnostic and therapeutic approaches. Nevertheless, further studies are necessary.

Rational design of soluble expressed human aldehyde dehydrogenase 2 with high stability and activity in pepsin and trypsin.

Acetaldehyde dehydrogenase 2 (ALDH2) is a crucial enzyme in alcohol metabolism, and oral administration of ALDH2 is a promising method for alcohol detoxification. However, recombinant ALDH2 is susceptible to hydrolysis by digestive enzymes in the gastrointestinal tract and is expressed as inactive inclusion bodies in E. coli. In this study, we performed three rounds of rational design to address these issues. Specifically, the surface digestive sites of pepsin and trypsin were replaced with other polar amino acids, while hydrophobic amino acids were incorporated to reshape the catalytic cavity of ALDH2. The resulting mutant DE2-852 exhibited a 45-fold increase in soluble expression levels, while its stability against trypsin and pepsin increased by eightfold and twofold, respectively. Its catalytic efficiency (kcat/Km) at pH 7.2 and 3.2 improved by more than four and five times, respectively, with increased Vmax and decreased Km values. The enhanced properties of DE2-852 were attributed to the D457Y mutation, which created a more compact protein structure and facilitated a faster collision between the substrate and catalytic residues. These results laid the foundation for the oral administration and mass preparation of highly active ALDH2 and offered insights into the oral application of other proteins.

LPA3 is a precise therapeutic target and potential biomarker for ovarian cancer.

Current studies have demonstrated that significant increased LPA levels to be observed in ascites in patients with ovarian cancer. Although several studies have shown that Lysophosphatidic acid (LPA) related to the progression of ovarian cancer, which LPA receptors (LPARs) and G-coupled protein subtypes mediated in LPA actions have not been clearly elucidated. This study aimed to clarify the roles of LPA and it is subtype-specific LPARs mediating mechanisms in ovarian cancer integrated using bioinformatic analysis and biological experimental approaches. The big data analysis shown that LPA3 was the only differentially expressed LPA receptor among the six LPARs in ovarian cancer and further verified in immunohistochemistry of tissue microarrays. Also found that LPA3 was also highly expressed in ovarian cancer tissue and ovarian cancer cells. Importantly, LPA significantly promoted the proliferation and migration of LPA3-overexpressing ovarian cancer cells, while the LPA-induced actions blocked by Ki16425, a LPAR1/3 antagonist treated, and LPA3-shRNA transfected. In vivo study indicated that the LPA3-overexpressing cell-derived tumors metastasis, tumors volume, and tumors mass were apparently increased in xenografted nude mice. In addition, we also observed that LPA3 was differential high expression in ovarian cancer tissue of the patients. Our studies further confirmed the LPA3/Gi/MAPKs/NF-κB signals were involved in LPA-induced oncogenic actions in ovarian cancer cells. Our findings indicated that the LPA3 might be a novel precise therapeutic target and potential biomarker for ovarian cancer.

The proliferative and anti-apoptosis functions of KGF/KGFR contributes to bronchial epithelial repair in asthma.

This study aimed to investigate the effect of keratinocyte growth factor (KGF) on the apoptosis, proliferation, damage repair, intercellular adhesion, and inflammatory cytokine release of cultured 16HBE 14o-bronchial ECs in vitro. Bronchial epithelial cells (ECs) from all subjects were obtained by bronchoscopic brushing. The expression levels of KGF and its receptor KGFR in collected cells were determined using RT-qPCR and Western blotting. The apoptosis and adhesion molecules expression by KGF administration were determined using flow cytometry and Western blotting. This occurred when 16HBE 14o-cell lines cultured and were exposed to interferon-γ (IFN-γ) and tumor necrosis factor-alpha (TNF-α) in vitro. The role of KGF on proliferation and damage repair were analyzed using CCK-8, EdU and wound closure assays after 16HBE 14o-cells were scraped. The effect of KGF on the release of inflammation related cytokines by damaged ECs was measured using ELISA kits. Compared with healthy controls, the KGF and KGFR expression and apoptosis significantly increased in collected cells from asthma patients. In vitro, treatment of KGF may limit IFN-γ and TNF-α induced apoptosis by inhibiting apoptosis-associated markers in the TNF signaling pathway. Besides, KGF could limit the release of TSLP, IL-25 and IL-33 by damaged 16HBE 14o-cells. On the contrary, KGF could promote the intercellular adhesion and wound closure of cultured 16HBE 14o-cells via the increased expression level of intercellular junction proteins ICAM-1, ß-catenin, E-cad, and Dsc3. In conclusion, KGF and KGFR may help bronchial ECs repair in asthma via the inhibition apoptosis of ECs while the promotion of proliferation and migration of ECs.

LPA receptor1 antagonists as anticancer agents suppress human lung tumours.

Lysophosphatidic acid (LPA), as a bioactive lipid, plays a variety of physiological and pathological roles via activating six types of G-protein-coupled LPA receptors (LPA1-6). Our preliminary study found that LPA1 is highly expressed in lung cancer tissues compared with paracancerous tissues, but the role of LPA1 in lung carcinoma is unclear. This study aimed to elucidate the association between LPA1 and lung tumour behaviour at the cellular and animal model levels. We found that LPA promoted the migration, proliferation and colony formation of a lung cancer cell line (A549). LPA1 and LPA3 are preferentially expressed in A549 cells, and both Ki16425 (LPA1 and LPA3 antagonist) and ono7300243 (LPA1 antagonist) completely blocked the LPA-induced actions. These results were further verified by experiments of the LPA1/3 overexpression and LPA1 knockdown A549 cells. Furthermore, LPA1 overexpression and knockdown A549 cells were used to assess the in vivo tumour-bearing animal model and the mechanism underlying LPA-induced actions. In the animal model, A549 cell-derived tumour volume was significantly increased by LPA1 overexpression and significantly decreased by LPA1 knockdown respectively, suggesting that LPA1 is a regulator of in vivo tumour formation. Our results also indicated that the LPA1/Gi/MAP kinase/NF-κB pathway is involved in LPA-induced oncogenic actions in A549 cells. Thus, targeting LPA1 may be a novel strategy for treating lung carcinoma.