Preventive effects of lactoferrin on acute alcohol-induced liver injury via iron chelation and regulation of iron metabolism.

Lactoferrin is widely found in milk and has the ability to bind iron. Previous studies have reported that lactoferrin was effective in the prevention and treatment of acute alcohol-induced liver injury (AALI). Ferroptosis is a recently discovered cell death and is involved in the development of AALI. However, the potential role of lactoferrin in acute alcohol-induced ferroptosis is still unclear. In this study, we observed that lactoferrin (10, 20, and 40 µg/mL) significantly mitigated alcohol (300 mM)-induced injury in vitro. Additionally, lactoferrin (100 and 200 mg/kg BW) significantly alleviated alcohol (4.8 g/kg BW)-induced injury in vivo. Our results showed that lactoferrin inhibited alcohol-induced upregulation of the ferroptosis marker protein ACSL4 and downregulation of GPX4. Meanwhile, lactoferrin treatment successfully reversed the elevated malondialdehyde (MDA) levels and the reduced glutathione (GSH) levels caused by alcohol treatment. These results may indicate that lactoferrin significantly decreased ferroptosis in vivo and in vitro. Lactoferrin has the potential to chelate iron, and our results showed that lactoferrin (20 µg/mL) significantly reduced iron ions and the expression of the ferritin heavy chain (FTH) under FeCl3 (100 µM) treatment. It was demonstrated that lactoferrin had a significant iron-chelating effect and reduced iron overload caused by FeCl3 in AML12 cells. Next, we examined iron content and the expression of iron metabolism marker proteins transferrin receptor (TFR), divalent metal transporter 1 (DMT1), FTH, and ferroportin (FPN). Our results showed that lactoferrin alleviated iron overload induced by acute alcohol. The expression of TFR and DMT1 was downregulated, and FPN and FTH were upregulated after lactoferrin treatment in vivo and in vitro. Above all, the study suggested that lactoferrin can alleviate AALI by mitigating acute alcohol-induced ferroptosis. Lactoferrin may offer new strategies for the prevention or treatment of AALI.

Sodium sulfite triggered hepatic apoptosis, necroptosis, and pyroptosis by inducing mitochondrial damage in mice and AML-12 cells.

Sodium sulfite (SS) is a biological derivative of the air pollutant sulfur dioxide, and is often used as a food and pharmaceutical additive. Improper or excessive SS exposure in liver cell death. The phenomenon of simultaneous regulation of apoptosis, necroptosis, and pyroptosis is defined as PANoptosis. However, the specific types of programmed cell death (PCD) caused by SS and their interconnections remain unclear. In the present study, C57BL/6 mice were orally administered SS for 30 d, consecutively, to establish an in vivo mouse exposure model. AML-12 cells were treated with SS for 24 h to establish an in vitro exposure model. The results showed that SS-induced mitochondrial reactive oxygen species (mtROS) accumulation activated the BAX/Bcl-2/caspase 3 pathway to trigger apoptosis and RIPK1/RIPK3/p-MLKL to trigger necroptosis. Interestingly, ROS-activated p-MLKL perforated not the cell membrane as well as the lysosomal membrane. We determined that p-MLKL mediates lysosomal membrane permeabilization (LMP), resulting in cathepsin B (CTSB) release. Furthermore, knockdown of MLKL, a CTSB inhibitor (CA074-ME) and an NLRP3 inhibitor (MCC950) alleviated SS-induced pyroptosis. In summary, our study showed that SS induced apoptosis and necroptosis though mtROS accumulation, whereas the activation of p-MLKL mediated NLRP3-dependent pyroptosis by causing CTSB leakage through LMP. This study comprehensively explored the mechanism unerlying SS-induced PCD and provided an experimental basis for p-MLKL as a potential regulatory protein in PANoptosis.

Alcohol induces hepatocytes necroptosis through the LC3/RIPK1/RIPK3 pathway.

Excessive alcohol consumption leads to serious liver injury. Necroptosis is a programmed cell death form, which has been confirmed to be involved in alcoholic liver injury. However, the exact mechanism remains still unclear. In this study, we found that ethanol caused hepatocytes necroptosis by activating receptor-interacting serine/threonine-protein kinase 1 and 3 (RIPK1 and RIPK3). Meanwhile, autophagy was activated in ethanol-treated hepatocytes. Accumulative studies have demonstrated a possible link between autophagy and necroptosis. Microtubule-associated protein 1 light chain 3 (LC3), an autophagy marker protein, is essential for autophagosome biogenesis/maturation. But little attention has been paid to its functional role. In this study, we explored whether LC3 was involved in ethanol-induced necroptosis. The data showed that LC3 interacted with RIPK1 and RIPK3 in ethanol-treated AML12 cells and mice liver by co-immunoprecipitation (co-IP) and colocalization assay. Ethanol-induced necrosome formation and subsequent necroptosis were alleviated in hepatocytes by knockdown of LC3 or autophagy inhibitor 3-methyladenine (3-MA). These results demonstrated that LC3 accumulation facilitated the formation of necrosome by LC3-RIPK1 and LC3-RIPK3 interactions, eventually caused hepatocytes necroptosis after acute ethanol exposure. Our current research could potentially offer a new understanding of the intricate mechanisms involved in the development of acute alcoholic liver injury.

The roles of FXYD family members in ovarian cancer: an integrated analysis by mining TCGA and GEO databases and functional validations.

BACKGROUND: The FXYD family of ion transport regulators have emerged as important modulators of cancer progression and metastasis. However, their expression and roles in ovarian cancer (OCa) have not been systematically investigated. METHODS: The expression of FXYD genes in OCa was analyzed using data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO), as well as independent clinical samples. The prognostic values of FXYD genes were evaluated by Kaplan-Meier and Cox regression analysis. To explore potential mechanisms, bioinformatics approaches including Gene Ontology, KEGG pathway analysis, GSEA and drug sensitivity correlation analysis were performed. OCa cell lines overexpressing FXYD1, FXYD5 or FXYD7 were also generated and their impacts on proliferation, migration and invasion were assessed. RESULTS: FXYD1 and FXYD6 were significantly downregulated while FXYD3, FXYD4 and FXYD5 were upregulated in OCa tissues compared to normal tissues. FXYD1, FXYD5 and FXYD7 were independent adverse prognostic factors for OCa patients. Pathway and drug correlation analysis revealed that FXYD1, FXYD5 and FXYD7 genes regulated diverse oncogenic signaling cascades and modulated the response to various chemotherapeutic agents. Overexpression of FXYD1, FXYD5 or FXYD7 enhanced OCa cell motility and invasiveness in vitro. CONCLUSION: Our results demonstrate aberrant expression patterns, prognostic values, and oncogenic activities of FXYD genes in OCa. FXYD1, FXYD5 and FXYD7 may serve as biomarkers and therapeutic targets for this disease. Targeting FXYD-mediated signaling represents a promising therapeutic strategy against OCa.

Sodium Sulfite-Triggered Hepatocyte Ferroptosis via mtROS/Lysosomal Membrane Permeabilization-Mediated Lysosome Iron Efflux.

Sodium sulfite is a widely used preservative in the food industry. Ferroptosis has been a newly discovered form of iron-dependent oxidative cell death in recent years. However, the potential connection between sodium sulfite and ferroptosis has not been explored. In our study, we observed the abnormal expression of ferroptosis marker protein in vivo, suggesting that sodium sulfite caused ferroptosis in vivo. Next, our study revealed that sodium sulfite caused the overproduction of mitochondrial reactive oxygen species (mtROS) in the AML-12 cells. It is well established that reactive oxygen species (ROS) can induce lysosomal membrane permeabilization. After lysosomal membrane permeabilization occurs, the outflow of Fe2+ in lysosomes triggers the Fenton reaction and subsequently results in the increase of intracellular ROS level, which is closely related to ferroptosis. As speculated, acridine orange (AO) staining and LysoTracker red staining showed that sodium sulfite-induced lysosomal membrane permeabilization could be alleviated by mtROS scavenger TEMPO. In addition, TEMPO, lysosomal stabilizer mannose, and lysosomal iron chelator deferoxamine (DFO) inhibited sodium sulfite-induced ferroptosis. Overall, the results showed that sodium sulfite induced lysosomal iron efflux through the mtROS-lysosomal membrane permeabilization pathway and eventually led to ferroptosis. Our study might provide a new mechanism for the hepatotoxicity of sodium sulfite and a theoretical basis for the risk assessment of sodium sulfite as a food additive.

Application of bone alkaline phosphatase and 25-oxhydryl-vitamin D in diagnosis and prediction of osteoporotic vertebral compression fractures.

BACKGROUND: Osteoporosis is a bone metabolic disease that usually causes fracture. The improvement of the clinical diagnostic efficiency of osteoporosis is of great significance for the prevention of fracture. The predictive and diagnostic values of bone alkaline phosphatase (B-ALP) and 25-oxhydryl-vitamin D (25-OH-VD) for osteoporotic vertebral compression fractures (OVCFs) were evaluated. METHODS: 110 OVCFs patients undergoing percutaneous vertebroplasty were included as subjects and their spinal computed tomography (CT) images were collected. After that, deep convolutional neural network model was employed for intelligent fracture recognition. Next, the patients were randomly enrolled into Ctrl group (65 cases receiving postoperative routine treatment) and VD2 group (65 cases injected with vitamin D2 into muscle after the surgery). In addition, 100 healthy people who participated in physical examination were included in Normal group. The differences in Oswestry dysfunction indexes (ODI), imaging parameters, B-ALP and 25-OH-VD expressions, and quality of life (QOL) scores of patients among the three groups were compared. The values of B-ALP and 25-OH-VD in predicting and diagnosing OVCFs and their correlation with bone density were analyzed. RESULTS: It was demonstrated that computer intelligent medical image technique was more efficient in fracture CT recognition than artificial recognition. In contrast to those among patients in Normal group, B-ALP rose while 25-OH-VD declined among patients in Ctrl and VD2 groups (P < 0.05). Versus those among patients in Ctrl group, ODI, Cobb angle, and B-ALP reduced, while bone density, the height ratio of the injured vertebrae, 25-OH-VD, and QOL score increased among patients in VD2 group after the treatment (P < 0.05). The critical values, accuracy, and areas under the curve (AUC) of the diagnosis of OVCFs by B-ALP and 25-OH-VD amounted to 87.8 µg/L versus 30.3 nmol/L, 86.7% versus 83.3%, and 0.86 versus 0.82, respectively. B-ALP was apparently negatively correlated with bone density (r = - 0.602, P < 0.05), while 25-OH-VD was remarkably positively correlated with bone density (r = 0.576, P < 0.05). CONCLUSION: To sum up, deep learning-based computer CT image intelligent detection technique could improve the diagnostic efficacy of fracture. B-ALP rose while 25-OH-VD declined among patients with OVCFs and OVCFs could be predicted and diagnosed based on B-ALP and 25-OH-VD. Postoperative intramuscular injection of VD2 could effectively improve the therapeutic effect on patients with OVCFs and QOL.

Alliin alleviates LPS-induced pyroptosis via promoting mitophagy in THP-1 macrophages and mice.

Pyroptosis is a new type of programmed cell death associated with inflammation. Excessive pyroptosis can cause body damage. Alliin is an organosulfur compound extracted from garlic, bearing anti-oxidation and anti-inflammatory properties. In this study, we revealed that alliin alleviated LPS-induced macrophage pyroptosis by detecting PI staining, IL-1ß and IL-18 release in vitro and in vivo. In the study of mechanism, we found that alliin might reduce the activation of NLRP3 inflammosome by decreasing intracellular ROS generation. Subsequently, we detected the effect of alliin on mitophagy which degraded damaged mitochondria. The results showed that alliin promoted PINK 1/Parkin-mediated mitophagy. After adding the mitophagy inhibitor CsA, the alleviating effect of alliin on mitochondrial damage and mitochondrial ROS were reversed and the relieving effect of alliin on LPS-induced pyroptosis was inhibited. These results suggested that alliin might reduce intracellular ROS production by promoting mitophagy, thus alleviating LPS-induced macrophages pyroptosis. Our study provides a new perspective and theoretical basis for alliin to alleviate pyroptosis which could further induce body damage.

Gas Phase Synthesis of the Elusive Trisilacyclopropyl Radical (Si3H5) via Unimolecular Decomposition of Chemically Activated Doublet Trisilapropyl Radicals (Si3H7).

The gas phase reaction of the simplest silicon-bearing radical silylidyne (SiH; X2Π) with disilane (Si2H6; X1A1g) was investigated in a crossed molecular beams machine. Combined with electronic structure calculations, our data reveal the synthesis of the previously elusive trisilacyclopropyl radical (Si3H5)-the isovalent counterpart of the cyclopropyl radical (C3H5)-along with molecular hydrogen via indirect scattering dynamics through long-lived, acyclic trisilapropyl (i-Si3H7) collision complex(es). Possible hydrogen-atom roaming on the doublet surface proceeds to molecular hydrogen loss accompanied by ring closure. The chemical dynamics are quite distinct from the isovalent methylidyne (CH)-ethane (C2H6) reaction, which leads to propylene (C3H6) radical plus atomic hydrogen but not to cyclopropyl (C3H5) radical plus molecular hydrogen. The identification of the trisilacyclopropyl radical (Si3H5) opens up preparative pathways for an unusual gas phase chemistry of previously inaccessible ring-strained (inorgano)silicon molecules as a result of single-collision events.

Associations between untraditional risk factors, pneumonia/lung cancer, and hospital fatality among hypertensive men in Guangzhou downtown.

Mortality of primary hypertension is high worldwide. Whether untraditional factors exist in modern life and affect the mortality is not well studied. The aim of the study was to evaluate the risk factors for fatality rate of hypertensive men in downtown area. A cross-sectional study was performed on hypertensive men, who were hospitalized into our hospital and lived in eligible urban areas. The characteristics of the patients and factors for the fatality were analyzed and of the risks or the contributors on the status were investigated. 14354 patients were identified. Mean age was 68.9 ± 12.4 year old (y) and dead ones was 75.9 ± 9.5 y. The overall hospitalized fatality was 5.9%, which was increased with age: fatality with 0.7%, 2.2%, 2.9%, 7.1%, 11.1% and 16.6% was for age group ≦ 49 y, 50-59 y, 60-69 y, 70-79 y, 80-89 y and ≧ 90 y respectively. The increased fatality was significantly positively correlated with the incidence of pneumonia, P < 0.05, r = 0.99. Pneumonia was prone to involve in men with older age and severer organ damage by hypertension. Similar to traditional risks such as coronary heart disease and stroke, pneumonia and lung cancer were also significantly associated with the fatality. Odds ratio (95% CI) for pneumonia and lung cancer were 6.18 (4.35-8.78) and 1.55 (1.14-2.11). The study provides evidence that pneumonia and lung cancer are highly associated with fatality of hypertensive men in downtown area, indicating that in order to reduce the fatality of hypertension, these lung diseases should be prevented and treated intensively in modern life.

N-Acetyl-l-cysteine/l-Cysteine-Functionalized Chitosan-ß-Lactoglobulin Self-Assembly Nanoparticles: A Promising Way for Oral Delivery of Hydrophilic and Hydrophobic Bioactive Compounds.

Self-assembled and cross-linked hybrid hydrogels for entrapment and delivery of hydrophilic and hydrophobic bioactive compounds were developed based on N-acetyl-l-cysteine (NAC)- or l-cysteine (CYS)-functionalized chitosan-ß-lactoglobulin nanoparticles (NPs). In both the systems, amphiphilic protein ß-lactoglobulin (ß-lg) was self-assembled by using glutaraldehyde for affinity binding with egg white-derived peptides (EWDP) and curcumin and then coated with NAC- or CYS-functionalized chitosan (CS) by electrostatic interaction. The resulting NPs were characterized in terms of size, polydispersity, and surface charge by dynamic light scattering. Results corroborated pH-sensitive properties of NAC-CS-ß-lg NPs and CYS-CS-ß-lg NPs with the particle size as small as 118 and 48 nm, respectively. The two kinds of NPs also showed excellent entrapment of EWDP and curcumin with the entrapment efficiency (EE) of EWDP and curcumin ranging from 51 to 89% and 42 to 57% in NAC-CS-ß-lg NPs, as well as 50-81% and 41-57% in CYS-CS-ß-lg NPs under different pH values. Fourier transform infrared and molecular docking studies provided support for the interaction mechanism of NAC/CYS-CS with ß-lg as well as the NPs with EWDP and curcumin. Strikingly, the in vitro release kinetics of EWDP and curcumin exhibited the controlled and sustained release properties up to 58 and 70 h from the NPs, respectively. Note that the permeability of QIGLF (pentapeptide, isolated from EWDP) and curcumin passing through Caco-2 cell monolayers were all improved after the entrapment in the NPs. This work offers promising methods for effective entrapment and oral delivery of both hydrophilic and hydrophobic bioactive compounds.

A vacuum ultraviolet photoionization study on the formation of methanimine (CH2NH) and ethylenediamine (NH2CH2CH2NH2) in low temperature interstellar model ices exposed to ionizing radiation.

Methylamine (CH3NH2) and methanimine (CH2NH) represent essential building blocks in the formation of amino acids in interstellar and cometary ices. In our study, by exploiting isomer selective detection of the reaction products via photoionization coupled with reflectron time of flight mass spectrometry (Re-TOF-MS), we elucidate the formation of methanimine and ethylenediamine (NH2CH2CH2NH2) in methylamine ices exposed to energetic electrons as a proxy for secondary electrons generated by energetic cosmic rays penetrating interstellar and cometary ices. Interestingly, the two products methanimine and ethylenediamine are isoelectronic to formaldehyde (H2CO) and ethylene glycol (HOCH2CH2OH), respectively. Their formation has been confirmed in interstellar ice analogs consisting of methanol (CH3OH) which is ioselectronic to methylamine. Both oxygen-bearing species formed in methanol have been detected in the interstellar medium (ISM), while for methanimine and ethylenediamine only methanimine has been identified so far. In comparison with the methanol ice products and our experimental findings, we predict that ethylenediamine should be detectable in these astronomical sources, where methylamine and methanimine are present.

Preparation of Antioxidant Peptides from Salmon Byproducts with Bacterial Extracellular Proteases.

Bacterial extracellular proteases from six strains of marine bacteria and seven strains of terrestrial bacteria were prepared through fermentation. Proteases were analyzed through substrate immersing zymography and used to hydrolyze the collagen and muscle proteins from a salmon skin byproduct, respectively. Collagen could be degraded much more easily than muscle protein, but it commonly showed weaker antioxidant capability. The hydrolysate of muscle proteins was prepared with crude enzymes from Pseudoalteromonas sp. SQN1 displayed the strongest activity of antioxidant in DPPH and hydroxyl radical scavenging assays (74.06% ± 1.14% and 69.71% ± 1.97%), but did not perform well in Fe2+ chelating assay. The antioxidant fractions were purified through ultrafiltration, cation exchange chromatography, and size exclusion chromatography gradually, and the final purified fraction U2-S2-I displayed strong activity of antioxidant in DPPH, hydroxyl radical scavenging assays (IC50 = 0.263 ± 0.018 mg/mL and 0.512 ± 0.055 mg/mL), and oxygen radical absorption capability assay (1.960 ± 0.381 mmol·TE/g). The final purified fraction U2-S2-I possessed the capability to protect plasmid DNA against the damage of hydroxyl radical and its effect was similar to that of the original hydrolysis product. It indicated that U2-S2-I might be the major active fraction of the hydrolysate. This study proved that bacterial extracellular proteases could be utilized in hydrolysis of a salmon byproduct. Compared with collagen, muscle proteins was an ideal material used as an enzymatic substrate to prepare antioxidant peptides.