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Human colon carcinomas, including HCT116 cells, often exhibit high basal autophagic flux under nutrient deprivation or hypoxic conditions. Mitochondrial ROS (mROS) is known as a “molecular switch”’ for regulating the autophagic pathway, which is critical for directing cancer cell survival or death. In early tumorigenesis, autophagy plays important roles in maintaining cellular homeostasis and contributes to tumor growth. However, the relationships between mROS and the autophagic capacities of HCT116 cells are poorly understood. Ubiquinol cytochrome c reductase binding protein (UQCRB) has been reported as a biomarker of colorectal cancer, but its role in tumor growth has not been clarified. Here, we showed that UQCRB was overexpressed in HCT116 cells compared to CCD18co cells, a normal colon fibroblast cell line. Pharmacological inhibition of UQCRB reduced mROS levels, autophagic flux, and the growth of HCT116 tumors in a xenograft mouse model. We further investigated mutant UQCRB-overexpressing cell lines to identify functional links in UQCRB-mROS-autophagy. Notably, an increasing level of mROS caused by UQCRB overexpression released Ca2+ by activation of lysosomal TRPML1 channels. This activation induced transcription factor EB nuclear translocation and lysosome biogenesis, leading to autophagy flux. Collectively, our study showed that increasing levels of mROS caused by overexpression of UQCRB in human colon carcinoma HCT116 cells could be linked to autophagy for cell survival.

Obesity is referred to as a condition in which excess body fat has accumulated to an extent that it causes negative impacts on health. The formation of body fat is regulated by complicated networks in relation to energy metabolism, and gut microbiota have been regarded as a key player. Studies have shown that supplements of probiotics provide benefits to health, including an improvement in metabolic syndrome and the control of body weight. In the present study, three probiotic strains, AP-32, bv-77, and CP-9, stood out from nine candidates using a lipid consumption assay, and were subsequently introduced to further animal tests. A rodent model of obesity was induced by a high-fat diet (HFD) in Sprague-Dawley (SD) rats, and three probiotic strains were administered either separately or in a mixture. A low dose (5 × 109 CFU/kg/day) and a high dose (2.5 × 1010 CFU/kg/day) of probiotics were orally provided to obese rats. The bioeffects of the probiotic supplements were evaluated based on five aspects: (1) the body weight and growth rate; (2) ketone bodies, non-esterified fatty acids (NEFAs), and feed efficiency; (3) blood biochemistry; (4) fat content; and (5) gut microbiota composition. Our results demonstrated that the supplement of AP-32, CP-9, and bv-77 alleviated the increasing rate of body weight and prevented the elevation of NEFAs and ketone bodies in obese rats. Although the effect on fat content showed a minor improvement, the supplement of probiotics displayed significant improvements in HFD-induced poor blood biochemical characteristics, such as alanine aminotransferase (ALT), aspartate Transaminase (AST), and uric acid, within 4 weeks. Furthermore, the combined supplement of three strains significantly increased Akkermansia mucinphila as compared with three individual strains, while its enrichment was negatively correlated with NEFAs and energy metabolism. In general, a mixture of three probiotic strains delivered a better outcome than a single strain, and the high dose of supplements provided a more profound benefit than the low dose. In conclusion, three probiotic strains, AP-32, bv-77, and CP-9, can alleviate body fat formation in obese rats. Furthermore, a combined supplement of these three probiotic strains may have potential in treating or controlling metabolic disorders.

Scyreprocin is an antimicrobial peptide first identified in the mud crab Scylla paramamosain. Herein, we showed that its recombinant product (rScyreprocin) could significantly inhibit the growth of human lung cancer NCI-H460 cells (H460), but showed no cytotoxicity to human lung fibroblasts (HFL1). rScyreprocin was a membrane-active peptide that firstly induced the generation of reactive oxygen species (ROS) in H460, and led to endoplasmic reticulum stress and Ca2+ release, which resulted in mitochondrial dysfunction and subsequently activation of caspase-3 cascades, and ultimately led to apoptosis. The comprehensive results indicated that rScyreprocin exerted anticancer activity by disrupting cell membrane and inducing apoptosis. The in vivo efficacy test demonstrated that intratumoral injection of rScyreprocin significantly inhibited the growth of H460 xenografts, which was close to that of the cisplatin (inhibition rate: 69.94% vs. 80.76%). Therefore, rScyreprocin is expected to become a promising candidate for the treatment of lung cancer.

Antimicrobial peptides (AMPs) identified in the reproductive system of animals have been widely studied for their antimicrobial activity, but only a few studies have focused on their physiological roles. Our previous studies have revealed the in vitro antimicrobial activity of two male gonadal AMPs, SCY2 and scyreprocin, from mud crab Scylla paramamosain. Their physiological functions, however, remain a mystery. In this study, the two AMPs were found co-localized on the sperm apical cap. Meanwhile, progesterone was confirmed to induce acrosome reaction (AR) of mud crab sperm in vitro, which intrigued us to explore the roles of the AMPs and progesterone in AR. Results showed that the specific antibody blockade of scyreprocin inhibited the progesterone-induced AR without affecting intracellular Ca2+ homeostasis, while the blockade of SCY2 hindered the influx of Ca2+. We further showed that SCY2 could directly bind to Ca2+. Moreover, progesterone failed to induce AR when either scyreprocin or SCY2 function was deprived. Taken together, scyreprocin and SCY2 played a dual role in reproductive immunity and sperm AR. To our knowledge, this is the first report on the direct involvement of AMPs in sperm AR, which would expand the current understanding of the roles of AMPs in reproduction.

Manipulating autophagy is a promising strategy for treating cancer as several autophagy inhibitors are shown to induce autophagic cell death. One of these, autophagonizer (APZ), induces apoptosis-independent cell death by binding an unknown target via an unknown mechanism. To identify APZ targets, we used a label-free drug affinity responsive target stability (DARTS) approach with a liquid chromatography/tandem mass spectrometry (LC–MS/MS) readout. Of 35 protein interactors, we identified Hsp70 as a key target protein of unmodified APZ in autophagy. Either APZ treatment or Hsp70 inhibition attenuates integrity of lysosomes, which leads to autophagic cell death exhibiting an excellent synergism with a clinical drug, temozolomide, in vitro, in vivo, and orthotropic glioma xenograft model. These findings demonstrate the potential of APZ to induce autophagic cell death and its development to combinational chemotherapeutic agent for glioma treatment. Collectively, our study demonstrated that APZ, a new autophagy inhibitor, can be used as a potent antitumor drug candidate to get over unassailable glioma and revealed a novel function of Hsp70 in lysosomal integrity regulation of autophagy.

Pubertal molt is a vital stage in the cultivation of mature female crabs in the aquacultural industry of Scylla paramamosain. Since fasting occurs during molting, which requires a large supply of energy, internal energy reserves are critical. However, the dynamics of energy supply during pubertal molt is not clear. This study focuses on the variations of carbohydrates and lipids in serum during the pubertal molt of S. paramamosain via a metabolomics approach. Eleven lipid or carbohydrate metabolic pathways were significantly influenced postmolt. A remarkable decrease in carbohydrates in serum suggested that free sugars were consumed for energy. A significant decrease in glucose and alpha-d-glucosamine 1-phosphate showed that chitin synthesis exhausted glycogen, resulting in insufficient glucose supply. An increase in l-carnitine and acetylcarnitine, and a significant decrease in 15 fatty acyls and 8 glycerophosphocholines in serum indicated that carnitine shuttle was stimulated, and β-oxidation was upregulated postmolt. In addition, astaxanthin, ponasterone A, and riboflavin in serum were significantly decreased postmolt. Eleven potential metabolite biomarkers were identified for pubertal molt. Taken together, carbohydrates and lipids were possibly major energy reserves in pubertal molt. This study suggests that an increase in carbohydrate and lipid levels in crab feed may alleviate the effects of fasting during molt and improve farm productivity in mature female crabs.

Background: Measuring the fraction of inspired oxygen (FiO2) is challenging in spontaneously breathing patients with impaired respiratory mechanics during low-flow nasal cannula. Our study investigates the FiO2 with varied tidal volume (VT) and respiratory rate (RR) among different lung mechanics and provides equations to estimate the FiO2. Methods: Two training and test lungs were used in this study, and the three lung mechanics (normal (R5/C60), restrictive (R20/C80), obstructive (R5/C40)) were designed. Spontaneous breathing with VT (300, 500, and 700 mL) and RR (10, 20, and 30 breaths/min) was simulated. The flow rate of the nasal cannula was set to 1, 3, and 5 L per minute (LPM), and the FiO2 was measured at the carina. Results: The lowest and highest FiO2 were evident during high (700 mL) and low VT (300 mL), respectively, among normal, restrictive, and obstructive lung models. As RR increases, this decreases the FiO2. However, we found that VT and oxygen flow rate are the principal factors influencing measured FiO2 by multiple linear regression analysis. Conclusions: Our data suggest that the actual FiO2 is never as high in spontaneously breathing patients as that estimated. VT and oxygen flow rate had a substantial impact on the FiO2.

This study is aimed at exploring the effects of multi-stage homogenization and trace amounts of Zr and Sc on the microstructures, mechanical properties, and recrystallization of Al&ndash, 4.5Zn&ndash, 1.5Mg alloys. The mechanical properties of the AA7005 aluminum alloys after the T6 heat treatment were evaluated through a hardness test and tensile test. The microstructures were analyzed by an optical microscope (OM), a differential scanning calorimeter (DSC), a transmission electron microscope (TEM), a scanning electron microscope (SEM), and electron backscattered diffraction (EBSD). The results show that the grain refinement effect of the as-cast, homogenized, and recrystallized Al&ndash, 1.5Mg alloy containing 0.05Sc (wt%) after the T6 heat treatment was more significant than that of the alloy containing 0.1Zr (wt%). In addition, compared with the aforementioned one-stage homogenization heat treatment, the two-stage homogenization made the dispersed grain phase (Al3Zr/Al3Sc) smaller. As a result, the T6 mechanical strength of the alloy after the two-stage homogenization heat treatment was better than that of the contrastive alloy after the one-stage homogenization heat treatment. However, the two different homogenization heat treatments caused a greater divergence between the sizes of the dispersed grain phases of the Al&ndash, 1.5Mg alloys containing Zr than between the sizes of the dispersed grain phases of the alloys containing Sc. Therefore, after the two-stage homogenization heat treatment, the alloy with 0.1Zr (wt%) promoted the mechanical properties better than the alloy with 0.05Sc (wt%).

The aim of this study is to explore the effects of Manganese addition and homogenization treatment on the microstructures and mechanical properties of the Al&ndash, 7Mg&ndash, 0.15Ti (B535.0) alloy. The optical microscopy, electrical conductivity measurements, transmission electron microscopy, scanning electron microscopy (SEM + EBSD), as well as Rockwell hardness and tensile tests, were exploited for this purpose. The main objectives are to refine the grain size, inhibit grain growth in the annealed state, and enhance the mechanical strength of the alloy. The results show that the addition of manganese to the Al&ndash, 0.15Ti alloys refined the as-cast and recrystallized grains of the alloys. During the homogenization process, Al4Mn high-temperature stable dispersoids were precipitated in the aluminum matrix. After annealing, the Al4Mn particles blocked the movement of grain boundaries during the growth of the recrystallized grains and inhibited grain growth. Consequently, the annealed alloys showed grain refinement and dispersion strengthening. The Al4Mn dispersoids of the alloys with manganese added were smaller and denser after a two-stage homogenization process compared to those that underwent a one-stage homogenization process. By contrast, for the alloys without the addition of manganese, the recrystallized grains showed normal growth after annealing, and different homogenization processes had no significantly different effects.

Antibiotics lead to increased susceptibility to colonization by pathogenic organisms, with different effects on the host-microbiota relationship. Here, we show that metronidazole treatment of specific pathogen-free (SPF) mice results in a significant increase of the bacterial phylum Proteobacteria in fecal pellets. Furthermore, metronidazole in SPF mice decreases hind limb muscle weight and results in smaller fibers in the tibialis anterior muscle. In the gastrocnemius muscle, metronidazole causes upregulation of Hdac4 , myogenin , MuRF1 , and atrogin1 , which are implicated in skeletal muscle neurogenic atrophy. Metronidazole in SPF mice also upregulates skeletal muscle FoxO3 , described as involved in apoptosis and muscle regeneration. Of note, alteration of the gut microbiota results in increased expression of the muscle core clock and effector genes Cry2 , Ror - β , and E4BP4 . PPARγ and one of its important target genes, adiponectin , are also upregulated by metronidazole. Metronidazole in germ-free (GF) mice increases the expression of other core clock genes, such as Bmal1 and Per2 , as well as the metabolic regulators FoxO1 and Pdk4 , suggesting a microbiota-independent pharmacologic effect. In conclusion, metronidazole in SPF mice results in skeletal muscle atrophy and changes the expression of genes involved in the muscle peripheral circadian rhythm machinery and metabolic regulation.

The effect of the addition of Be and Sc on the microstructure and mechanical properties of A357 alloy were systematically investigated. The results show that the addition of small amounts of Be and Sc could change the acicular structure of iron-bearing intermetallic compounds to harmless compact Al-Fe-Si and Sc-Fe iron-bearing intermetallic compounds. Compact iron-bearing intermetallic compounds could improve fluidity, causing a reduction in interdendritic shrinkage during solidification. The addition of 0.05 wt % Be enhanced the quality index of the A357 alloy by 11% and increased the notch-yield ratio of fracture toughness by 4.5%. In contrast, the addition of 0.05 wt % Sc increased the quality index and the notch to yield ratio of fracture toughness up to 17% and 9%, respectively. Therefore, the microstructure and mechanical properties of the A357 alloy could be improved by substituting Be with Sc.

Polygonatum alte-lobatum Hayata, a rhizomatous perennial herb, belongs to the Liliaceae family and is endemic to Taiwan. We investigated the antioxidant and anti-fatigue activities of P. alte-lobatum in exercised rats. Levels of polyphenols, flavonoids and polysaccharides and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical scavenging activity were measured in extracts of P. alte-lobatum (EPA). Sprague-Dawley rats were randomly divided into four groups for 8-week treatment with vehicle (control) and low-, medium-, and high-dose EPA (LEPA, MEPA, HEPA, 0, 75, 150, and 375 mg/kg/day, respectively). Exercise performance was evaluated by exhaustive treadmill exercise time and by changes in body composition and biochemical variables at the end of the experiment. EPA contained polyphenols, flavonoids and polysaccharides, with polysaccharide content at least 26 times greater than that of polyphenols and flavonoids. Trend analysis revealed that EPA dose-dependently scavenged DPPH free radicals. EPA treatment dose-dependently increased endurance running time to exhaustion and superoxide dismutase activity and total antioxidant ability of blood. EPA dose-dependently decreased serum urea nitrogen and malondialdehyde levels after exercise. Hepatic glycogen content, an important energy source for exercise, was significantly increased with EPA treatment. EPA could be a potential agent with an anti-fatigue pharmacological function.

Neuronal cell death in the central nervous system has always been a challenging process to decipher. In normal physiological conditions, neuronal cell death is restricted in the adult brain, even in aged individuals. However, in the pathological conditions of various neurodegenerative diseases, cell death and shrinkage in a specific region of the brain represent a fundamental pathological feature across different neurodegenerative diseases. In this review, we will briefly go through the general pathways of cell death and describe evidence for cell death in the context of individual common neurodegenerative diseases, discussing our current understanding of cell death by connecting with renowned pathogenic proteins, including Tau, amyloid-beta, alpha-synuclein, huntingtin and TDP-43.