Surface-tuning TiO2 NR photoanodes using CoOx interlayers and NiFe-LDH cocatalysts for photoelectrochemical wastewater treatment.

Increasing multidrug-resistant pathogenic microbial around the world become a global problem, making it imperative to develop effective methods for bacterial inactivation in wastewater. In this study, we propose a multifunctional photoelectrochemical (PEC) system to successfully disinfect microbial cells and degrade orange (II) dyes. CoOx NP were synthesized by spin-coating onto hydrothermally synthesized TiO2 nanorod arrays followed by electrodeposited NiFe-LDH to develop the NiFe-LDH/CoOx NP-TiO2 NRs. Interestingly, spin-coated CoOx NP-TiO2 NRs exhibited a 1.5-fold enhancement in photocurrent (∼1.384 mA/cm2) than pristine TiO2 NRs (0.92 mA/cm2). A NiFe-layered double hydroxide (LDH) cocatalysts layer further exhibits the maximum photocurrent density of 1.64 mA/cm2 with IPCE of 84.5% at 1.0 VAg/AgCl at 380 nm. Furthermore, NiFe-LDH/CoOx-TiO2 NR photoanodes were effectually employed for photoelectrochemical bacteria disinfection and organic pollutant removals. With NiFe-LDH/CoOx-TiO2 NR, 99% (120 min) bacterial inactivation and 99% (60 min) orange II dye decomposition efficiency was achieved. Superoxide radicals (O2-), hydroxyl radicals (OH), and holes (h+) played a critical role in the PEC degradation systems. Due to the synergy between NiFe-LDH cocatalyst and CoOx interlayer, surface water oxidation reactions were accelerated over NiFe-LDH/CoOx NP-TiO2 NRs. The charge transport process in NiFe-LDH/CoOx NP-TiO2 NRs photoanode-based PEC system was proposed in detail.

In-situ Hf/Zr co-doped Fe2O3 nanorod decorated with CuOx/CoOx: Enhanced photocatalytic performance for antibacterial and organic pollutants.

Herein, we successfully synthesized Hf/Zr co-doping on Fe2O3 nanorod photocatalyst by a hydrothermal process and quenching methods. The synergistic roles of Hf and Zr double-doping on the bacteria inactivation test and decomposition of organic pollutants were investigated in detail for the 1 wt% CoOx loaded Hf/Zr-Fe2O3 NRs and CuOx/CoOx loaded Hf/Zr-Fe2O3 NRs photocatalyst. Initially, the rod-like porous morphology of the Hf/Zr-doped Fe2O3 NRs was produced via a hydrothermal method at various Hf co-doping (0, 2, 4, 7 and 10)%. Further, CoOx and CuOx loaded by a wet impregnation approach on the Hf/Zr-Fe2O3 NRs and a highly photoactive Hf(4)/Zr-Fe2O3 [CoOx/CuOx] NRs photocatalyst were developed. After the Hf(4)/Zr-Fe2O3 [CoOx/CuOx] NRs photocatalyst treatment, the Bio-TEM imagery of bacterial cells showed extensive morphological deviations in cell membranes. Hf(4)/Zr-Fe2O3 NR achieved 84.1% orange II degradation upon 3 h illumination, which is higher than that of Hf-Fe2O3 and Zr-Fe2O3 (68.7 and 73.5%, respectively). Additionally, the optimum sample, Hf(4)/Zr-Fe2O3 [CoOx/CuOx] photocatalyst, exhibited 95.5% orange II dye degradation after light radiation for 3 h. Optimized Hf(4)/Zr-Fe2O3 [CoOx/CuOx] catalysts exhibited 99.9% and 99.7% inactivation of E. coli and S. aureus with 120 min, respectively. Further, scavenger experiments revealed that the electrons are the primary responsible species for photocatalytic kinetics. This work will provide a rapid method for the development of high photocatalytic performance materials for bacterial disinfection and organic degradation.

Bio-electrochemical degradation of carbamazepine (CBZ): A comprehensive study on effectiveness, degradation pathway, and toxicological assessment.

Recent attention on the detrimental effects of pharmaceutically active compounds (PhACs) in natural water has spurred researchers to develop advanced wastewater treatment methods. Carbamazepine (CBZ), a widely recognized anticonvulsant, has often been a primary focus in numerous studies due to its prevalence and resistance to breaking down. This study aims to explore the effectiveness of a bio-electrochemical system in breaking down CBZ in polluted water and to assess the potential harmful effects of the treated wastewater. The results revealed bio-electro degradation process demonstrated a collaborative effect, achieving the highest CBZ degradation compared to electrodegradation and biodegradation techniques. Notably, a maximum CBZ degradation efficiency of 92.01% was attained using the bio-electrochemical system under specific conditions: Initial CBZ concentration of 60 mg/L, pH level at 7, 0.5% (v/v) inoculum dose, and an applied potential of 10 mV. The degradation pathway established by identifying intermediate products via High-Performance Liquid Chromatography-Mass Spectrometry, revealed the complete breakdown of CBZ without any toxic intermediates or end products. This finding was further validated through in vitro and in vivo toxicity assays, confirming the absence of harmful remnants after the degradation process.

Evaluation of Lentilactobacillus parafarraginis A6-2 strain for aluminum removal and anti-inflammatory effects: implications for alleviating Al toxicity.

AIM: To assess the effectiveness of Lentilactobacillus parafarraginis A6-2 cell lysate for the removal of aluminum (Al), which induces neurotoxicity, and its protective effect at cellular level. METHODS AND RESULTS: The cell lysate of the selected L. parafarraginis A6-2 strain demonstrated superior Al removal compared to live or dead cells. The Al removal efficiency of L. parafarraginis A6-2 cell lysate increased with decreasing pH and increasing temperature, primarily through adsorption onto peptidoglycan. Neurotoxicity mitigation potential of L. parafarraginis A6-2 was evaluated using C6 glioma cells. C6 cells exposed with increasing concentration of Al led to elevated toxicity and inflammation, which were gradually alleviated upon treatment with L. parafarraginis A6-2. Moreover, Al-induced oxidative stress in C6 cells showed a concentration-dependent reduction upon treatment with L. parafarraginis A6-2. CONCLUSIONS: This study demonstrated that L. parafarraginis A6-2 strain, particularly in its lysate form, exhibited enhanced capability for Al removal. Furthermore, it effectively mitigated Al-induced toxicity, inflammation, and oxidative stress.

Lactiplantibacillus sp. D10-2: potential bacteria for eliminating bisphenol A and reducing BpA-induced lipid accumulation.

Bisphenol A (BpA) is an endocrine-disrupting substance commonly found in plastics and resins. It is reported that BpA exposure induces lipid accumulation in humans, similar to obesogenic compounds. The main objective of this study is to investigate the removal of BpA using Lactiplantibacillus sp. D10-2, and to examine its potential for reducing BpA-induced lipid accumulation in 3T3-L1 cell line model. The heat-dried cells of Lactiplantibacillus sp. D10-2 showed 69.7% removal efficiency for initial BpA concentration of 10 µg/mL, which was 30.5% higher than the live cells. The absence of metabolites or intermediates in BpA removal studies indicates that the Lactiplantibacillus sp. D10-2 strain removed BpA by adsorption process. The hydrophobic interactions of heat-dried Lactiplantibacillus sp. D10-2 cells were observed to be higher with 33.7% compared to live cells (15.0%), suggesting a stronger ability to bind with BpA. Although the BpA binding onto Lactiplantibacillus sp. D10-2 was not affected by pH, it was confirmed that as the temperature increases, the binding ability got decreased due to mass transfer and diffusion of BpA molecules. Treatment with Lactiplantibacillus sp. D10-2 (0.1, 0.25, 0.5, 1%) reduced lipid accumulation by 61.7, 58.0, 52.7 and 60.4% in 3T3-L1 cells exposed with BpA. In addition, it was confirmed that Lactiplantibacillus sp. D10-2 treatment suppressed the protein expression levels of lipogenesis-related PPARγ and C/EBPα in 3T3-L1 cells. The results of the study suggest that the Lactiplantibacillus sp. D10-2 strain can remove BpA and reduce BpA-accelerated lipid accumulation in 3T3-L1 cells.

Applied potential assisted biodegradation of amoxicillin (AMX) using bacterial consortium isolated from a waste dump site.

Antibiotics have revolutionized modern day living with their ability to effectively treat infectious diseases in humans and animals. However, the release of antibiotic compounds into the environment has led to toxic consequences. To reduce this environmental impact, it is important to employ an inexpensive and rational technology to reduce the amount of antibiotics released into the ecosystem. This study aims to explore the potential of using a bio-electrochemical system (BES) to remove Amoxicillin (AMX) from artificially contaminated soil using a microbial consortium and pure culture isolates. Under desired conditions, including an initial AMX concentration of 150 mg/L, 5 mg/L tryptone as the nitrogen source, pH of 7, temperature of 29 °C, an applied potential of 0.8 V, and an inoculum dose of 1% w/v, the BES showed a maximum degradation of 97.9% of AMX with the microbial consortium (HP03, HP09, and HP10). High performance liquid chromatography-mass spectrometry was used to analyse the intermediates formed during the degradation process, and the pathway elucidated revealed complete degradation of AMX. Phytotoxicity studies and degradation efficiency against multiple antibiotics confirmed the environmental significance of the BES with microbial consortium. Overall, this study highlights the potential of BES as a cost-effective and efficient method for reducing the release of antibiotics into the environment and provides valuable insights into the mechanisms and pathways of antibiotic degradation.

Effect of fermentation on antioxidant, antimicrobial, anti-inflammatory, and anti-Helicobacter pylori adhesion activity of Ulmus davidiana var. japonica root bark.

The limited yield of Ulmus davidiana var. japonica root bark (URB) extract is considered an economic loss to the food industry. Improving extraction yield and bioactivity through fermentation increase the industrial usage of URB. The study aims to optimize the fermentation with cellulolytic and pectinolytic bacteria and evaluate the bioactivity and anti-Helicobacter pylori activity of the fermented URB extract. URB fermentation with the Bacillus licheniformis FLa3, isolated from salted seafood (Sardinella zunasi), under optimal conditions (37 °C, pH 6, 10% inoculum dose, and 36 h) improved the extraction yield by 36% compared to the control. The antioxidant and antimicrobial activity of the fermented extract were significantly higher than non-fermented extract. High-performance liquid chromatography results confirmed that the fermentation increased the proportion of bioactive components such as catechin (171.7%), epicatechin (144.3%), quercetin (27.3%), and kaempferol (16.7%). The results confirmed that the fermentation increased both the extraction yield and bioactivity.

Erysiphe lonicerigena sp. nov., a Powdery Mildew Species Found on Lonicera harae.

A powdery mildew (Erysiphaceae) has been continuously collected on the leaves of Lonicera harae in the southern part of the Korean Peninsula, where this shrub is indigenous. Microscopic examination of the asexual morphs revealed that the current collections are differentiated from the all known Erysiphe species on Lonicera spp. by its longer conidiophores and longer conidia. Although the morphology of the chasmothecia is reminiscent of Erysiphe ehrenbergii and E. lonicerae, the specimens on L. harae differ from them in having smaller ascospores. A phylogenetic tree generated from a combined dataset of the internal transcribed spacer region and 28S rDNA gene sequences demonstrates that sequences obtained from three powdery mildew collections on L. harae clustered together as an independent species clade with high bootstrap values distant from other Erysiphe species on Lonicera, representing a species of its own. Based on morphological differences and molecular-phylogenetic results, the powdery mildew on L. harae is proposed as a new species, Erysiphe lonicerigena, and the holomorph of the fungus is described and illustrated in this study.

A combination of BR101801 and venetoclax enhances antitumor effect in DLBCL cells via c-Myc/Bcl-2/Mcl-1 triple targeting.

Double hit diffuse large B-cell lymphoma (DLBCL) with rearrangement and overexpression of both c-Myc and Bcl-2 responds poorly to standard R-CHOP therapy. In a recent phase I study, Venetoclax (ABT-199) targeting Bcl-2 also exhibited disappointing response rates in patients with relapsed/refractory DLBCL, suggesting that targeting only Bcl-2 is not sufficient for achieving successful efficacy due to the concurrent oncogenic function of c-Myc expression and drug resistance following an increase in Mcl-1. Therefore, co-targeting c-Myc and Mcl-1 could be a key combinatorial strategy to enhance the efficacy of Venetoclax. In this study, BR101801 a novel drug for DLBCL, effectively inhibited DLBCL cell growth/proliferation, induced cell cycle arrest, and markedly inhibited G0/G1 arrest. The apoptotic effect of BR101801 was also observed by increased Cytochrome C, cleaved PARP, and Annexin V-positive cell populations. This anti-cancer effect of BR101801 was confirmed in animal models, where it effectively inhibited tumor growth by reducing the expression of both c-Myc and Mcl-1. Furthermore, BR101801 exhibited a significant synergistic antitumor effect even in late xenograft models when combined with Venetoclax. Our data strongly suggest that c-Myc/Bcl-2/Mcl-1 triple targeting through a combination of BR101801 and Venetoclax could be a potential clinical option for double-hit DLBCL.

Microwave-assisted thermochemical conversion of Zr-FeOOH nanorods to Zr-ZnFe2O4 nanorods for bacterial disinfection and photo-Fenton catalytic degradation of organic pollutants.

Herein, we report a CoOx-loaded Zr-doped ZnFe2O4 (CoOx/Zr-ZFO) NR photocatalyst synthesized by successive microwave and wet impregnation methods for bacterial inactivation and degradation of organic pollutants. For the first time, microwave treatment was used for Zn attachment on hydrothermally synthesized self-assembled Zr-FeOOH NRs to produce Zr-doped ZnFe2O4 (Zr-ZFO) NRs. The lowest bandgap energy (1.96 eV) enables for significant absorption in the visible light region, which helps to improve bacteria degradation inactivation efficiency. Further, various metal oxides (Cu, Ag and Co) were loaded onto the surface of photocatalysts (Zr-ZFO NRs) by a wet impregnation method. As-synthesized CoOx/Zr-ZFO-3 NRs were systematically characterized and used as photocatalysts for inactivation of E. coli and S. aureus and degradation of organic pollutants. The CoOx/Zr-ZFO-3 NR photocatalyst exhibited better inactivation efficiency (99.4 %) than other metal oxide-loaded Zr-ZFO NRs (Ag2Ox-loaded Zr-ZFO NRs (33.6 %), CuOx-loaded Zr-ZFO NRs (77.6 %)). Additionally, the optimum CoOx/Zr-ZFO-3 NR photocatalyst showed 98.3 % and 98.1 % degradation efficiencies for BPA and orange II dye, respectively, under visible light irradiation (λ ≥ 420 nm). Therefore, this work affords a novel, simple and rapid approach for the development of photocatalysts which active in visible light for bacterial disinfection and organic degradation.

Crystal Structure of H227A Mutant of Arginine Kinase in Daphnia magna Suggests the Importance of Its Stability.

Arginine kinase (AK) plays a crucial role in the survival of Daphnia magna, a water flea and a common planktonic invertebrate sensitive to water pollution, owing to the production of bioenergy. AK from D. magna (DmAK) has four highly conserved histidine residues, namely, H90, H227, H284, and H315 in the amino acid sequence. In contrast to DmAK WT (wild type), the enzyme activity of the H227A mutant decreases by 18%. To identify the structure-function relationship of this H227A mutant enzyme, the crystal 3D X-ray structure has been determined and an unfolding assay using anilino-1-naphthalenesulfonic acid (ANS) fluorescence has been undertaken. The results revealed that when compared to the DmAK WT, the hydrogen bonding between H227 and A135 was broken in the H227A crystal structure. This suggests that H227 residue, closed to the arginine binding site, plays an important role in maintaining the structural stability and maximizing the enzyme activity through hydrogen bonding with the backbone oxygen of A135.

Combination Therapy of the Active KRAS-Targeting Antibody inRas37 and a PI3K Inhibitor in Pancreatic Cancer.

KRAS activating mutations, which are present in more than 90% of pancreatic cancers, drive tumor dependency on the RAS/mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K)/AKT signaling pathways. Therefore, combined targeting of RAS/MAPK and PI3K/AKT signaling pathways may be required for optimal therapeutic effect in pancreatic cancer. However, the therapeutic efficacy of combined MAPK and PI3K/AKT signaling target inhibitors is unsatisfactory in pancreatic cancer treatment, because it is often accompanied by MAPK pathway reactivation by PI3K/AKT inhibitor. Therefore, we developed an inRas37 antibody, which directly targets the intra-cellularly activated GTP-bound form of oncogenic RAS mutation and investigated its synergistic effect in the presence of the PI3K inhibitor BEZ-235 in pancreatic cancer. In this study, inRas37 remarkably increased the drug response of BEZ-235 to pancreatic cancer cells by inhibiting MAPK reactivation. Moreover, the co-treatment synergistically inhibited cell proliferation, migration, and invasion and exhibited synergistic anticancer activity by inhibiting the MAPK and PI3K pathways. The combined administration of inRas37and BEZ-235 significantly inhibited tumor growth in mouse models. Our results demonstrated that inRas37 synergistically increased the antitumor activity of BEZ-235 by inhibiting MAPK reactivation, suggesting that inRas37 and BEZ-235 co-treatment could be a potential treatment approach for pancreatic cancer patients with KRAS mutations.

Elucidation of the electron transfer environment in the MMOR FAD-binding domain from Methylosinus sporium 5.

By facilitating electron transfer to the hydroxylase diiron center, MMOR-a reductase-serves as an essential component of the catalytic cycle of soluble methane monooxygenase. Here, the X-ray structure analysis of the FAD-binding domain of MMOR identified crucial residues and its influence on the catalytic cycle.

Antimicrobial and Antibiofilm Effect of ε-Polylysine against Salmonella Enteritidis, Listeria monocytogenes, and Escherichia coli in Tryptic Soy Broth and Chicken Juice.

ε-Polylysine (ε-PL) is a safe food additive that is used in the food industry globally. This study evaluated the antimicrobial and antibiofilm activity of antibacterial peptides (ε-PL) against food poisoning pathogens detected in chicken (Salmonella Enteritidis, Listeria monocytogenes, and Escherichia coli). The results showed that minimum inhibitory concentrations (MICs) ranged between 0.031-1.0 mg/mL, although most bacterial groups (75%) showed MICs of 1.0 mg/mL. The reduction in the cell viability of pathogens due to ε-PL depended on the time and concentration, and 1/2 × MIC of ε-PL killed 99.99% of pathogens after 10 h of incubation. To confirm biofilm inhibition and degradation effects, crystal violet assay and confocal laser scanning microscopy (CLSM) were used. The biofilm formation rates of four bacterial groups (Salmonella, Listeria, E. coli, and multi-species bacteria) were 10.36%, 9.10%, 17.44%, and 21.37% at 1/2 × MIC of ε-PL, respectively. Additionally, when observed under a CLSM, ε-PL was found to induce biofilm destruction and bacterial cytotoxicity. These results demonstrated that ε-PL has the potential to be used as an antibiotic and antibiofilm material for chicken meat processing.

Enhanced Ultraviolet Photoresponse Characteristics of Indium Gallium Zinc Oxide Photo-Thin-Film Transistors Enabled by Surface Functionalization of Biomaterials for Real-Time Ultraviolet Monitoring.

Indium gallium zinc oxide (IGZO) is one of the most promising materials for diverse optoelectronic applications based on thin-film transistors (TFTs) including ultraviolet (UV) photodetectors. In particular, the monitoring of UV-A (320-400 nm) exposure is very useful for healthcare applications because it can be used to prevent various human skin and eye-related diseases. However, the relatively weak optical absorption in the UV-A range and the persistent photoconductivity (PPC) arising from the oxygen vacancy-related states of IGZO thin films limit efficient UV monitoring. In this paper, we report the enhancement of the UV photoresponse characteristics of IGZO photo-TFTs by the surface functionalization of biomolecules on an IGZO channel. The biomaterial/IGZO interface plays a crucial role in enhancing UV-A absorption and suppressing PPC under negative gate bias, resulting in not only increased photoresponsivity and specific detectivity but also a fast and repeatable UV photoresponse. In addition, turning off the device without external bias completely eliminates PPC due to the internal electric field induced by the surface functionalization of biomaterials. Such a volatile feature of PPC enables the fast and repeatable UV photoresponse. These results suggest the potential of IGZO photo-TFTs combined with biomaterials for real-time UV monitoring.

ANGPTL4 accelerates KRASG12D-Induced acinar to ductal metaplasia and pancreatic carcinogenesis.

Oncogenic KRASG12D induces neoplastic transformation of pancreatic acinar cells through acinar-to-ductal metaplasia (ADM) and pancreatic intraepithelial neoplasia (PanIN), and drives pancreatic ductal adenocarcinoma (PDAC). Angiopoietin-like 4 (ANGPTL4) is known to be involved in the regulation of cancer growth and metastasis. However, whether ANGPTL4 affects KRASG12D-mediated ADM and early PDAC intervention remains unknown. In the current study, we investigated the role of ANGPTL4 in KRASG12D-induced ADM, PanIN formation, and PDAC maintenance. We found that ANGPTL4 was highly expressed in human and mouse ADM lesions and contributed to the promotion of KRASG12D-driven ADM in mice. Consistently, ANGPTL4 rapidly induced ADM in three-dimensional culture of acinar cells with KRAS mutation and formed ductal cysts that silenced acinar genes and activated ductal genes, which are characteristic of in vivo ADM/PanIN lesions. We also found that periostin works as a downstream regulator of ANGPTL4-mediated ADM/PDAC. Genetic ablation of periostin diminished the ADM/PanIN phenotype induced by ANGPTL4. A high correlation between ANGPTL4 and periostin was confirmed in human samples. These results demonstrate that ANGPTL4 is critical for ADM/PanIN initiation and PDAC progression through the regulation of periostin. Thus, the ANGPTL4/periostin axis is considered a potential target for ADM-derived PDAC.

Experiences of Pathology Course among Hospital Management Graduates.

The purpose of this study was to explore hospital management graduates' experience in pathology courses. Data were gathered through four focus group interviews by 16 hospital management graduates who attended pathology courses. Data were collected from June to August, 2020. Conventional content analysis was used for data analysis. Six categories were extracted that described hospital management graduates' experience in pathology courses, as follows: "Suggestions for the curriculum," "Students' preference for pathology professor," "Demands for various teaching methods," "Broad and difficult class content," "Recognition of pathology courses during college years," and "The importance of studying the pathology course realized after graduation." The findings suggest that it is important to identify hospital management graduates' perspectives to improve pathology curriculum in the educational process. Additionally, it is necessary to continuously connect educational and practical environments for the effective management of pathology courses.

Intracellular KRAS-specific antibody enhances the anti-tumor efficacy of gemcitabine in pancreatic cancer by inducing endosomal escape.

KRAS mutation is associated with the progression and growth of pancreatic cancer and contributes to chemo-resistance, which poses a significant clinical challenge in pancreatic cancer. Here, we developed a RT22-ep59 antibody (Ab) that directly targets the intracellularly activated GTP-bound form of oncogenic KRAS mutants after it is internalized into cytosol by endocytosis through tumor-associated receptor of extracellular epithelial cell adhesion molecule (EpCAM) and investigated its synergistic anticancer effects in the presence of gemcitabine in pancreatic cancer. We first observed that RT22-ep59 specifically recognized tumor-associated EpCAM and reached the cytosol by endosomal escape. In addition, the anticancer effect of RT22-ep59 was observed in the high-EpCAM-expressing pancreatic cancer cells and gemcitabine-resistant pancreatic cancer cells, but it had little effect on the low-EpCAM-expressing pancreatic cancer cells. Additionally, co-treatment with RT22-ep59 and gemcitabine synergistically inhibited cell viability, migration, and invasion in 3D-cultures and exhibited synergistic anticancer activity by inhibiting the RAF/ERK or PI3K/AKT pathways in cells with high-EpCAM expression. In an orthotopic mouse model, combined administration of RT22-ep59 and gemcitabine significantly inhibited tumor growth. Furthermore, the co-treatment suppressed cancer metastasis by blocking EMT signaling in vitro and in vivo. Our results demonstrated that RT22-ep59 synergistically increased the antitumor activity of gemcitabine by inhibiting RAS signaling by specifically targeting KRAS. This indicates that co-treatment with RT22-ep59 and gemcitabine might be considered a potential therapeutic strategy for pancreatic cancer patients harboring KRAS mutation.

Insight into Structural Aspects of Histidine 284 of Daphnia magna Arginine Kinase.

Arginine kinase (AK), a bioenergy-related enzyme, is distributed widely in invertebrates. The role of highly conserved histidines in AKs is still unascertained. In this study, the highly conserved histidine 284 (H284) in AK of Daphnia magna (DmAK) was replaced with alanine to elucidate the role of H284. We examined the alteration of catalytic activity and structural changes of H284A in DmAK. The catalytic activity of H284A was reduced dramatically compared to that in wild type (WT). Thus the crystal structure of H284A displayed several structural changes, including the alteration of D324, a hydrogen-bonding network around H284, and the disruption of π-stacking between the imidazole group of the H284 residue and the adenine ring of ATP. These findings suggest that such alterations might affect a conformational change of the specific loop consisting of G310-V322 at the antiparallel ß-sheet region. Thus, we speculated that the H284 residue might play an important role in the conformational change of the specific loop when ATP binds to the substrate-binding site of DmAK.

ANGPTL4 exacerbates pancreatitis by augmenting acinar cell injury through upregulation of C5a.

Pancreatitis is the inflammation of the pancreas. However, little is known about the genes associated with pancreatitis severity. Our microarray analysis of pancreatic tissues from mild and severe acute pancreatitis mice models identified angiopoietin-like 4 (ANGPTL4) as one of the most significantly upregulated genes. Clinically, ANGPTL4 expression was also increased in the serum and pancreatic tissues of pancreatitis patients. The deficiency in ANGPTL4 in mice, either by gene deletion or neutralizing antibody, mitigated pancreatitis-associated pathological outcomes. Conversely, exogenous ANGPTL4 exacerbated pancreatic injury with elevated cytokine levels and apoptotic cell death. High ANGPTL4 enhanced macrophage activation and infiltration into the pancreas, which increased complement component 5a (C5a) level through PI3K/AKT signaling. The activation of the C5a receptor led to hypercytokinemia that accelerated acinar cell damage and furthered pancreatitis. Indeed, C5a neutralizing antibody decreased inflammatory response in LPS-activated macrophages and alleviated pancreatitis severity. In agreement, there was a significant positive correlation between C5a and ANGPTL4 levels in pancreatitis patients. Taken together, our study suggests that targeting ANGPTL4 is a potential strategy for the treatment of pancreatitis.