The molecular mechanisms regulating the assembly of the autophagy initiation complex.

The autophagy initiation complex is brought about via a highly ordered and stepwise assembly process. Two crucial signaling molecules, mTORC1 and AMPK, orchestrate this assembly by phosphorylating/dephosphorylating autophagy-related proteins. Activation of Atg1 followed by recruitment of both Atg9 vesicles and the PI3K complex I to the PAS (phagophore assembly site) are particularly crucial steps in its formation. Ypt1, a small Rab GTPase in yeast cells, also plays an essential role in the formation of the autophagy initiation complex through multiple regulatory pathways. In this review, our primary focus is to discuss how signaling molecules initiate the assembly of the autophagy initiation complex, and highlight the significant roles of Ypt1 in this process. We end by addressing issues that need future clarification.

Energy deprivation-induced autophagy and aggrephagy: insights from yeast and mammals. / ä»Žé µæ¯å’Œå“ºä¹³åŠ¨ç‰©ç»†èƒžè§£æžèšé›†ä½“è‡ªå™¬å’Œèƒ½é‡åŒ®ä¹è¯±å¯¼çš„è‡ªå™¬.

Autophagy plays a crucial role in maintaining cellular homeostasis in response to various stimuli. Compared to research on nutrient deprivation-induced autophagy, the understanding of the molecular mechanisms and physiological/pathological significance of autophagy triggered by energy deprivation remains limited. A primary focus of our lab is to elucidate how cells sense energy deprivation and initiate autophagy. Using the model organisms Saccharomyces cerevisiae and mammalian cells, we found that cellular reactive oxygen species (ROS), DNA damage sensor Mec1, and mitochondrial aerobic respiration play essential roles in the autophagy induced by energy deprivation. This review aims to provide a concise overview of these research findings.

Research and development of fully enclosed wire-shell support structure technology for deep soft rock roadway based on TRIZ theory.

The TRIZ theory was used to accurately discover the problems to be solved in the design of roadway surrounding rock control technology. This paper tried to solve the complex issue of surrounding rock control in deep roadways from a new perspective. Based on the functional component analysis and causal axis analysis of the problem's primary reason, simultaneously, the surrounding rock control technology was optimized through technical contradiction analysis, physical contradiction analysis, and substance and field model analysis. As a result, a fully enclosed wire-shell support technology was proposed. Finally, taking the typical soft rock roadway engineering of Pansan Coal Mine in Huainan Mining Area, Anhui Province, China, as the engineering background, the engineering application and effect evaluation were completed. This paper provides a reference for controlling the instability of deep soft rock roadways in coal mines. A new idea of optimizing roadway support engineering based on TRIZ theory was proposed.

TOR-mediated Ypt1 phosphorylation regulates autophagy initiation complex assembly.

The regulation of autophagy initiation is a key step in autophagosome biogenesis. However, our understanding of the molecular mechanisms underlying the stepwise assembly of ATG proteins during this process remains incomplete. The Rab GTPase Ypt1/Rab1 is recognized as an essential autophagy regulator. Here, we identify Atg23 and Atg17 as binding partners of Ypt1, with their direct interaction proving crucial for the stepwise assembly of autophagy initiation complexes. Disruption of Ypt1-Atg23 binding results in significantly reduced Atg9 interactions with Atg11, Atg13, and Atg17, thus preventing the recruitment of Atg9 vesicles to the phagophore assembly site (PAS). Likewise, Ypt1-Atg17 binding contributes to the PAS recruitment of Ypt1 and Atg1. Importantly, we found that Ypt1 is phosphorylated by TOR at the Ser174 residue. Converting this residue to alanine blocks Ypt1 phosphorylation by TOR and enhances autophagy. Conversely, the Ypt1S174D phosphorylation mimic impairs both PAS recruitment and activation of Atg1, thus inhibiting subsequent autophagy. Thus, we propose TOR-mediated Ypt1 as a multifunctional assembly factor that controls autophagy initiation via its regulation of the stepwise assembly of ATG proteins.

Atg1-mediated Atg11 phosphorylation is required for selective autophagy by regulating its association with receptor proteins.

Atg11 is an adaptor protein required for the induction of selective autophagy via receptor binding. However, our understanding of the molecular mechanisms by which it regulates selective autophagy remains incomplete. Here, we show that Atg11 is phosphorylated by Atg1. Rapamycin treatment or starvation conditions induced slower electrophoretic mobility of Atg11 in an Atg1 kinase activity-dependent manner. Through in vitro kinase assays combined with mutagenesis, we determined that Atg1 phosphorylates S949, S1057, and S1064 residues in CC4 domain of Atg11. Replacing the three residues with alanine suppressed the cleavage of selective autophagy substrates for the cytoplasm-to-vacuole targeting (Cvt) pathway, mitophagy, reticulophagy, and pexophagy. The Atg11 mutant was defective in binding to related selective autophagy receptors. These results demonstrate a previously unknown function of Atg1 in regulation of selective autophagy via Atg11 phosphorylation.Abbreviations: AMPK: AMP-activated protein kinase; ATG: autophagy-related; Cvt: cytoplasm-to-vacuole targeting; FUNDC1: FUN14 domain-containing protein 1; GFP: green fluorescent protein; MTOR: mechanistic target of rapamycin kinase; PAS: phagophore assembly site; PIK3C3: phosphatidylinositol 3-kinase catalytic subunit type 3; PRKAC/PKA: protein kinase cAMP-activated; SD-G: glucose starvation; SD-N: nitrogen starvation; ULK1: unc-51 like autophagy activating kinase 1; λ-PPase: lambda protein phosphatase.

Mec1 regulates PAS recruitment of Atg13 via direct binding with Atg13 during glucose starvation-induced autophagy.

Mec1 is a DNA damage sensor, which performs an essential role in the DNA damage response pathway and glucose starvation-induced autophagy. However, the functions of Mec1 in autophagy remain unclear. In response to glucose starvation, Mec1 forms puncta, which are recruited to mitochondria through the adaptor protein Ggc1. Here, we show that Mec1 puncta also contact the phagophore assembly site (PAS) via direct binding with Atg13. Functional analysis of the Atg13-Mec1 interaction revealed two previously unrecognized protein regions, the Mec1-Binding Region (MBR) on Atg13 and the Atg13-Binding Region (ABR) on Mec1, which mediate their mutual association under glucose starvation conditions. Disruption of the MBR or ABR impairs the recruitment of Mec1 puncta and Atg13 to the PAS, consequently blocking glucose starvation-induced autophagy. Additionally, the MBR and ABR regions are also crucial for DNA damage-induced autophagy. We thus propose that Mec1 regulates glucose starvation-induced autophagy by controlling Atg13 recruitment to the PAS.

Research on Damage Evolution Law of Glazed Hollow Beads-Cement/Sodium Silicate Grouting Materials under Different Cycles of Loading and Unloading.

With the depletion of shallow resources, deep resource mining has become a trend. However, the high temperature and complex stress environment in deep mines make resource extraction extremely challenging. This paper developed a thermal insulation grouting material made of glazed hollow beads, sodium silicate, and cement and tested the compressive strength, gelation time, and stone rate under various curing days in light of the issue of high temperature heat damage in high ground temperature mines and the impact of mining on roadway grouting bolt support. Fatigue strength, fatigue deformation, load-residual strain, energy evolution and microscopic features were studied and analyzed in relation to the damage law of graded cyclic loading and unloading under the number of varying cycles. The findings demonstrate that cyclic loading and unloading strength is lower than uniaxial compressive strength. The fatigue strength is significantly decreased when the number of cycles reaches its limit. Residual strain is less sensitive to changes in stress than load strain. The fitting correlation coefficients of total output energy and elastic energy are higher than 0.71.

Performance degradation and damage model of rice husk ash concrete under dry-wet cycles of sulfate environment.

Rice husk ash concrete (RHAC) is a new type of concrete that has been rapidly gaining acceptance in recent years. In this paper, the improvement effect of rice husk ash (RHA) on the sulfate erosion performance of concrete was confirmed. The ratio of rice husk ash concrete (RHAC) was optimized and compared with ordinary concrete (OC). The performance degradation of 9%RHAC (rice husk ash at 9% by weight of cement) and OC within 135 times erosion dry-wet cycles solution with Na2SO4 at 5% by weight of solution were studied, including the change of apparent phenomena, compressive strength, tensile strength, effective porosity, and dynamic elastic modulus. The microstructure changes of samples before and after sulfate dry-wet cycle were observed by using a scanning electron microscope (SEM). The results show that with the increase of sulfate dry-wet cycle times, the concrete specimen gradually peels off and expands in volume. The compressive strength and tensile strength increase first and then drop sharply, the effective porosity decreases first and then increases, and the relative dynamic elastic modulus increases and then decreases. The reason is that the ettringite and gypsum are formed by the reaction of sulfate intrusion and hydration products under wetting treatment. After drying treatment, ettringite and free water combine to form sodium sulfate. In the early of circulation, ettringite, gypsum, and sodium sulfate fill the internal pores of the concrete and improve the density. As the number of sulfate dry-wet cycles increases, expansion products accumulate, causing structural expansion damage and deterioration of mechanical performance. However, the hydrated calcium silicate hydrate gel was produced by mixing rice husk ash with concrete to improve the material strength and corrosion resistance. The deterioration degree of the 9%RHAC is better than that of OC at all stages. Finally, the damage constitutive models were established, and the accuracy is higher compared with the measured value.

A syringe-aided apta-nanosensing method for colorimetric determination of acetamiprid.

A syringe-aided apta-nanosensing method is reported for the colorimetric determination of acetamiprid. The method employs double-stranded (ds) DNA-conjugated gold nanoparticle@magnetic agarose beads, i.e., dsDNA-AuNP@MABs as peroxidase-mimicking composite probes, in which the aptamer is indirectly attached to the AuNP surface through its hybridization with complementary DNA (cDNA). Upon contact with the acetamiprid target, the probes can give perceptible color change due to the possible conformation switch from dsDNA's brush-like to cDNA's 'pancake' regime. An "air-spaced pumping" procedure using a syringe equipped with ring magnets as the operation platform was proposed to facilitate the magnetic separation of the sensing probes. Therefore, the analytical steps can be easily accomplished in a syringe, including probe loading, acetamiprid capture and magnetic separation from crude samples, chromogenic reagent loading and colorimetric visualization. Acetamiprid concentration down to 3.3 ppb can be easily identified by the naked eye. The final solution also can be transferred for quantitative measurement. Under spectrometer, the ratio of the absorbance at 652 nm in the presence and absence of acetamiprid (A/A0) is linearly related to the acetamiprid concentration in the 0.4-4.5 ppb range. The limit of detection is calculated to be 0.24 ppb. Moreover, satisfactory recoveries ranging from 90.90 to 91.82% with relative standard deviations of ≤2.96% were obtained in analyzing real spiked samples.

Effects of High Temperature on Creep Behaviour of Glazed Hollow Bead Insulation Concrete.

Glazed hollow bead insulation concrete (GHBC) presents a promising application prospect in terms of its light weight and superior fire resistance. However, only a few studies have focused on the creep behaviour of GHBC exposed to high temperatures. Therefore, in this study, the mechanism of high temperature on GHBC is analysed through a series of tests on uniaxial compression and multistage creep of GHBC, exposed from room temperature up to 800 °C. The results show a decrease in the weight and compressive strength of GHBC as the temperature rises. After 800 °C, the loss of weight and strength reach to 9.67% and 69.84%, respectively. The creep strain and creep rate increase, with a higher target temperature and higher stress level, while the transient deformation modulus, the creep failure threshold stress, and creep duration are reduced significantly. Furthermore, the creep of GHBC exhibits a considerable increase above 600 °C and the creep under the same loading ratio at 600 °C increases by 74.19% compared to the creep at room temperature. Indeed, the higher the temperature, the more sensitive the stress is to the creep. Based on our findings, the Burgers model agrees well with the creep test data at the primary creep and steady-state creep stages, providing a useful reference for the fire resistance design calculation of the GHBC structures.

Performance Degradation and Microscopic Analysis of Lightweight Aggregate Concrete after Exposure to High Temperature.

This study analyses the deterioration of mechanical properties in lightweight concrete after exposure to room temperature (20 C) and high temperature, i.e., up to 1000 C, including changes in visual appearance, loss of mass, and compressive strength. All-lightweight shale ceramsite aggregate concrete (ALWAC) and semi-lightweight shale ceramsite aggregate concrete (SLWAC) are prepared using an absolute volume method to analyse the relationships between relative ultrasonic pulse velocity, loss rate of compressive strength, damage degree, and temperature levels. Our results show that, under high temperature, the lightweight aggregate ceramsite concrete performs better compared to normal concrete. After exposure to 1000 C, the ALWAC shows a strength loss of no more than 80%, while the normal concrete loses its bearing capacity, with a similar strength loss as the SLWAC. Furthermore, the relative ultrasonic pulse velocity and damage degree are used to evaluate the effects of high temperature on the concretes, including the voids and cracks on the surface and inside of the specimens, which induces the deterioration of mechanical properties and contributes to the thermal decomposition of the cementing system and the loss of cohesion at the aggregate interface. Based on internal structure analyses, the results from this study confirm that the lightweight aggregate concrete shows a high residual compressive strength after exposure to the high temperature.

Mitochondrial Fusion Machinery Specifically Involved in Energy Deprivation-Induced Autophagy.

Mitochondria are highly dynamic organelles, which can form a network in cells through fusion, fission, and tubulation. Its morphology is closely related to the function of mitochondria. The damaged mitochondria can be removed by mitophagy. However, the relationship between mitochondrial morphology and non-selective autophagy is not fully understood. We found that mitochondrial fusion machinery, not fission or tubulation machinery, is essential for energy deprivation-induced autophagy. In response to glucose starvation, deletion of mitochondrial fusion proteins severely impaired the association of Atg1/ULK1 with Atg13, and then affected the recruitment of Atg1 and other autophagic proteins to PAS (phagophore assembly site). Furthermore, the deletion of fusion proteins blocks mitochondrial respiration, the binding of Snf1-Mec1, the phosphorylation of Mec1 by Snf1, and the dissociation of Mec1 from mitochondria under prolonged starvation. We propose that mitochondrial fusion machinery regulates energy deprivation-induced autophagy through maintaining mitochondrial respiration.

Atg11 is required for initiation of glucose starvation-induced autophagy.

How energy deprivation induces macroautophagy/autophagy is not fully understood. Here, we show that Atg11, a receptor protein for cargo recognition in selective autophagy, is required for the initiation of glucose starvation-induced autophagy. Upon glucose starvation, Atg11 facilitates the interaction between Snf1 and Atg1, thus is required for Snf1-dependent Atg1 activation. Phagophore assembly site (PAS) formation requires Atg11 via its control of the association of Atg17 with Atg29-Atg31. The binding of Atg11 with Atg9 is crucial for recruiting Atg9 vesicles to the PAS and, thus, glucose starvation-induced autophagy. We propose Atg11 as a key initiation factor controlling multiple key steps in energy deprivation-induced autophagy. Abbreviations: AMPK: AMP-activated protein kinase; Ams1: α-mannosidase; Ape1: aminopeptidase I; Cvt: cytoplasm-to-vacuole targeting; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; GFP: green fluorescent protein; MBP: myelin basic protein; MMS: methanesulfonate; PAS: phagophore assembly site; PNBM: p-nitrobenzyl mesylate; SD-G: glucose starvation medium; SD-N: nitrogen starvation medium; ULK1, unc-51 like autophagy activating kinase 1; WT: wild type.

Printing the Ultra-Long Ag Nanowires Inks onto the Flexible Textile Substrate for Stretchable Electronics.

: Printing technology offers a simple and cost-effective opportunity to develop all-printed stretchable circuits and electronic devices, possibly providing ubiquitous, low-cost, and flexible devices. To successfully prepare high-aspect-ratio Ag nanowires (NWs), we used water and anhydrous ethanol as the solvent and polyvinylpyrrolidone (PVP) as the viscosity regulator to obtain a water-soluble Ag NWs conductive ink with good printability. Flexible and stretchable fabric electrodes were directly fabricated through screen printing. After curing at room temperature, the sheet resistance of the Ag NW fabric electrode was 1.5 Ω/sq. Under a tensile strain of 0-80% and with 20% strains applied for 200 cycles, good conductivity was maintained, which was attributed to the inherent flexibility of the Ag NWs and the intrinsic structure of the interlocked texture.

An aptamer based aggregation assay for the neonicotinoid insecticide acetamiprid using fluorescent upconversion nanoparticles and DNA functionalized gold nanoparticles.

An acetamiprid-binding aptamer (ABA), gold nanoparticles (AuNPs) and upconversion nanoparticles (UCNPs) are used in a colorimetric and fluorometric method for the ultrasensitive and selective detection of the pesticide acetamiprid. The ABA is first configured into a duplex with a complementary DNA covalently attached to AuNPs. The resulting dsDNA-functionalized AuNP probe is not stable in 0.15 M NaCl solution and aggregates. This causing the color to change from red to purple. In the presence of acetamiprid, the ABA undergoes a structural switch from a DNA duplex to an aptamer-acetamiprid complex and consequently dissociates from the AuNPs. The partially unhybridized AuNPs are stable against salt-induced aggregation and show red color. The ratio of absorbances at 524 nm (red) and 650 nm (purple blue) varies with the concentration of acetamiprid in the 0.025-10 µM concentration range. The colorimetric signal can be further amplified by introducing DNA-modified carboxylated UCNPs (silica-coated NaYF4:Yb,Er) which display red and green fluorescence under 980 nm excitation. An inner filter effect occurs between DNA-modified UCNPs and dsDNA-modified AuNPs. The fluorometric assay is based on the measurement of the ratio of red (654 nm) and green (540 nm) fluorescence and works in the 0.025 to 1 µM acetamiprid concentration range and has a 0.36 nM detection limit (at a signal-to-noise ratio of 3). Because of the specificity of the aptamer, the assay is high selective. It was successfully used to quantify acetamiprid in contaminated real samples. Graphical abstract Schematic presentation of an upconversion fluorescent assay for acetamiprid. It involves the principle of analyte-triggered structural switch of aptamers, salt-induced AuNP aggregation, and signal amplification from UCNP.

NIR light-activated upconversion semiconductor photocatalysts.

Harvesting of near infrared (NIR) light in the abundant and environmentally friendly solar spectrum is particularly significant to enhance the utilization rate of the cleanest energy on earth. Appreciating the unique nonlinear optical properties of upconversion materials for converting low-energy incident light into high-energy radiation, they become the most promising candidates for fabricating NIR light-active photocatalytic systems by integrating with semiconductors. The present review summarizes recent NIR light-active photocatalytic systems based on a sequence of NaYF4-based, fluoride-based, oxide-based and Ln3+ ion-doped semiconductor-based photocatalysts for degradation of organic molecules. In addition, we provide an in-depth analysis of various photocatalytic mechanisms and enhancement effects for efficient photo-redox performance of different upconversion semiconductor photocatalysts. We envision that this review can inspire multidisciplinary research interest in rational design and fabrication of efficient full-spectrum active (UV-visible-NIR) photocatalytic systems and their wider applications in solar energy conversion.

Tunable and ultra-stable UV light-switchable fluorescent composites for information hiding and storage.

Advanced fluorescent materials have demonstrated great value in the anti-counterfeiting field for information storage and hiding. In this work, a novel strategy for producing UV light-switchable fluorescent patterns to hide and store information is proposed based on carbon dots (CDs) and Y2O3:Eu composites with exceptional optical properties. CDs can present blue emission under both 254 nm and 365 nm UV light excitation, but Y2O3:Eu can only arouse a red color under 254 nm light excitation. Under conventional 365 nm excitation, a single blue color and partial information are shown, however, the whole information in patterns can be displayed under specific 254 nm excitation. The functional anti-counterfeiting patterns are made by screen printing using as-prepared polyvinyl alcohol (PVA)-medium fluorescent inks. Moreover, the hidden information retained its integrity after printed patterns were exposed to an ambient environment for 90 days. Such invisible, tunable and ultra-stable fluorescent patterns utilizing 254 nm UV light well achieve information hiding and storage and increase information security for anti-counterfeiting applications.

Acid-degradable lactobionic acid-modified soy protein nanogels crosslinked by ortho ester linkage for efficient antitumor in vivo.

It remains a crucial challenge to achieve efficient cellular uptake and intracellular drug release in tumor cells for the nanoscale drug delivery systems. Herein, acid-degradable nanogels were prepared by cross-linking methacrylated soy protein with an acid-labile ortho ester cross-linker (NG1), and then modified with lactobionic acid (LA) to give tumor-targeted nanogels (NG2). Both NG1 and NG2 displayed excellent stability in neutral environment, while showed pH-triggered degradation behaviors under mildly acidic conditions resulting from the breakage of ortho ester bonds. Doxorubicin (DOX) was successfully loaded into nanogels, which exhibited an accelerated release at low pH. In vitro cell studies demonstrated that LA-modified nanogels could effectively improve cellular internalization, show higher cytotoxicity and apoptosis toward asialoglycoprotein receptor (ASGPR) over-expressed HepG2 cells. In vivo antitumor experimentproved that LA modification could significantly enhance the tumor-targeting ability of nanogels and increase DOX concentration in tumor site, leading to better therapeutic efficacy. Histological analysis further demonstrated that soy protein-based nanogels did not cause any damage to normal organs. Overall, these pH-sensitive and tumor-targeting soy protein-based nanogels can be potential drug carriers for efficient tumor treatment.

Low molecular weight polyethylenimine-grafted soybean protein gene carriers with low cytotoxicity and greatly improved transfection in vitro.

A series of gene carriers (SP-PEI) have been synthesized by acylation reaction between soybean protein and branched polyethylenimine with low molecular weight of 600, 1200 and 1800 Da, and designed as SP-PEI600, SP-PEI1200 and SP-PEI1800, respectively. SP-PEI could effectively condense plasmid DNA into nanoscale polyplexes with size range of 100-200 nm, and exhibited much lower cytotoxicity against 293T and SH-SY5Y cells than that of branched polyethylenimine (25 kDa). In vitro gene transfection demonstrated that SP-PEI/DNA complex displayed increased transfection against 293T and SH-SY5Y cells with the increase of the weight ratio of SP-PEI/DNA complex with or without 10% serum. At weight ratio of 24, SP-PEI1800/DNA polyplexes showed the highest transfection on SH-SY5Y cells, which was almost three folds higher than PEI (25 kDa). Furthermore, these SP-PEIs/DNA polyplexes could effectively transfect 293T and SH-SY5Y cells with or without 10% serum, suggesting their excellent serum tolerance.