α-lipoic acid ameliorates consequences of copper overload by up-regulating selenoproteins and decreasing redox misbalance.

α-lipoic acid (LA) is an essential cofactor for mitochondrial dehydrogenases and is required for cell growth, metabolic fuel production, and antioxidant defense. In vitro, LA binds copper (Cu) with high affinity and as an endogenous membrane permeable metabolite could be advantageous in mitigating the consequences of Cu overload in human diseases. We tested this hypothesis in 3T3-L1 preadipocytes with inactivated Cu transporter Atp7a; these cells accumulate Cu and show morphologic changes and mitochondria impairment. Treatment with LA corrected the morphology of Atp7a-/- cells similar to the Cu chelator bathocuproinedisulfonate (BCS) and improved mitochondria function; however, the mechanisms of LA and BCS action were different. Unlike BCS, LA did not decrease intracellular Cu but instead increased selenium levels that were low in Atp7a-/- cells. Proteome analysis confirmed distinct cell responses to these compounds and identified upregulation of selenoproteins as the major effect of LA on preadipocytes. Upregulation of selenoproteins was associated with an improved GSH:GSSG ratio in cellular compartments, which was lowered by elevated Cu, and reversal of protein oxidation. Thus, LA diminishes toxic effects of elevated Cu by improving cellular redox environment. We also show that selenium levels are decreased in tissues of a Wilson disease animal model, especially in the liver, making LA an attractive candidate for supplemental treatment of this disease.

Synthesis of Coumarin-Based Photosensitizers for Enhanced Antibacterial Type I/II Photodynamic Therapy.

Photodynamic therapy (PDT) is an effective method for treating microbial infections by leveraging the unique photophysical properties of photosensitizing agents, but issues such as fluorescence quenching and the restricted generation of reactive oxygen species (ROS) under hypoxic conditions still remain. In this study, we successfully synthesized and designed a coumarin-based aggregation-induced emission luminogen (AIEgen), called ICM, that shows a remarkable capacity for type I ROS and type II ROS generation. The 1O2 yield of ICM is 0.839. The ROS it produces include hydroxyl radicals (HO•) and superoxide anions (O2•-), with highly effective antibacterial properties specifically targeting Staphylococcus aureus (a Gram-positive bacterium). Furthermore, ICM enables broad-spectrum fluorescence imaging and exhibits excellent biocompatibility. Consequently, ICM, as a potent type I photosensitizer for eliminating pathogenic microorganisms, represents a promising tool in addressing the threat posed by these pathogens.

Stunning methods before slaughter induce oxidation changes of large yellow croaker during cold storage: the role of mitochondria and underlying mechanisms.

BACKGROUND: Improper stunning methods before slaughter could cause fish to deteriorate more quickly during cold storage. However, it is unclear how stunning methods affect the mitochondrial structure and the role of mitochondria in oxidation in muscle-based food. RESULTS: This study explored the potential mechanism of oxidation induced by different stunning methods (hit on the head, T1 ; gill cut, T2 ; immersion in ice/water slurry, T3 ; CO2 asphyxiated, T4 ; 40% CO2 + 30% N2 + 30% O2 , T5 ) in large yellow croaker during cold storage. The results showed that T4 samples had the minimum stress response and the mitochondrial membrane potential and permeability were less damaged. Besides, the mitochondrial functional structure and peroxisome of T4 samples were less damaged compared with other samples, which was reflected in higher total superoxide dismutase, catalase and glutathione peroxidase activities. In terms of oxidation indices, the T4 samples showed higher pH values and iron myoglobin contents and lower total volatile basic nitrogen and thiobarbituric acid reactive substances after 168 h cold storage, indicating that the T4 samples significantly maintained oxidative stability of large yellow croaker. CONCLUSION: The CO2 asphyxiation had the least oxidative damage to large yellow croaker during cold storage, possibly because it had the least effect on mitochondrial structure, reactive oxygen species and antioxidant enzyme activity. © 2023 Society of Chemical Industry.

Larkinella punicea sp. nov., isolated from manganese mine soil.

Strain ZZJ9T is a Gram-stain-negative, rod-shaped, aerobic bacterium isolated from manganese mine soil. Strain ZZJ9T showed the highest 16S rRNA gene sequence similarities with Larkinella rosea 15J16-1T3AT (97.1%), Larkinella terrae 15J8-8T (97.0%), Larkinella knui 15J6-3T6T (96.8%), and Larkinella ripae 15J11-1T (95.3%). The genome size of strain ZZJ9T was 8.01 Mb and the DNA G+C content was 51.8 mol%. ANI values among strain ZZJ9T and Larkinella rosea 52004 T, Larkinella knui KCTC 42998T, and Larkinella terrae 52001T were 80.5%, 82.7%, and 80.5%, respectively. dDDH values among strain ZZJ9T and Larkinella rosea 52004T, Larkinella knui KCTC 42998T, and Larkinella terrae 52001T were 23.5%, 26.0%, and 23.6%, respectively. Furthermore, the genome of strain ZZJ9T contained 6302 predicted protein-coding genes and 3114 (49%) of them had classificatory functions. The major quinone of strain ZZJ9T was menaquinone-7 and the main cellular fatty acids were C16:1ω5c (39.5%), iso-C15:0 (25.6%), and iso-C17:0 3OH (11.5%). The polar lipids of strain ZZJ9T were phosphatidylethanolamine, unidentified lipid, and two unidentified aminolipids. Based on the results of phylogenetic, genome, phenotypic, and chemotaxonomic analytical, strain ZZJ9T represents a novel species of the genus Larkinella, for which the name Larkinella punicea sp. Nov. is proposed. The type strain is ZZJ9T (= KCTC 62876T = CCTCC AB 2018215T).

Rubusoside-assisted solubilization of poorly soluble C6-Ceramide for a pilot pharmacokinetic study.

Although C6-Ceramide has attracted much attention as a possible tumor suppressor, the delivery of C6-Ceramide is still challenging due to its inherent hydrophobicity and insolubility. In this study we explored the use of a natural compound rubusoside (RUB) as a solubilizer to enhance the solubility of a fluorescence-labeled C6-Ceramide (NBD C6-Ceramide) and to characterize its pharmacokinetics and tissue distribution in an animal model. RUB significantly enhanced the solubility of NBD C6-Ceramide by forming nanomicelles, and efficiently delivered NBD C6-Ceramide in rats by oral and intravenous administration. RUB loaded 1.96 % of NBD C6-Ceramide in the nanomicelles and solubilized it to a concentration of 3.6 mg/mL in water. NBD C6-Ceramide in nanomicelles remained stable in aqueous solutions, allowing intravenous administration without the use of any organic solvents or surfactants. After oral administration, NBD C6-Ceramide rapidly rose to peak plasma concentrations within the first 90 min, distributed to tissues, and remained in vivo for more than 24 h. Tissular levels of NBD C6-Ceramide from high to low were associated with heart, lung, cerebellum, testicle, spleen, liver, kidney, and brain. Altogether, our study demonstrated that RUB-assisted nanomicelles can serve as an efficient and convenient delivery system for short-chain C6-Ceramide and enable in vivo evaluation of potential new cancer treatments.

Mucilaginibacter terrenus sp. nov., isolated from manganese mine soil.

Strain ZH6T is a Gram-stain-negative, rod-shaped, aerobic bacterium isolated from manganese mine soil. Strain ZH6T had highest 16S rRNA gene sequence similarities to Mucilaginibacter yixingensis YX-36T (96.9 %) and Mucilaginibacter psychrotolerans NH7-4T (96.8 %). The genome size of strain ZH6T was 4.61 Mb with a DNA G+C content of 44.0 mol%. The average nucleotide identity and digital DNA-DNA hybridization values between strain ZH6T and M. yixingensis DSM 26809T were 70.6 and 19.2 %, respectively. Strain ZH6T had menaquinone-7 as a major quinone and main cellular fatty acids of iso-C15 : 0, iso-C17 : 0 3-OH and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c). The polar lipids of strain ZH6T were a phosphatidylethanolamine, an unidentified glycolipid, an unidentified phospholipid, three unidentified aminophospholipids and four unidentified lipids. Based on the phenotypic, chemotaxonomic and phylogenetic results, strain ZH6T represents a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacterterrenus sp. nov., is proposed. The type strain is ZH6T (=CCTCC AB 2018373T=KCTC 72075T).

Silk fibroin nanoparticles for enhanced bio-macromolecule delivery to the retina.

The aim of this study was to investigate intravitreal injection of silk fibroin nanoparticles (SFNs) encapsulating bio-macromolecules, achieving enhanced drug bioavailability, and extended retention in retina. SFNs were prepared with regenerated silk fibroin using desolvation method with fluorescein isothiocyanate labeled bovine serum albumin (FITC-BSA) as bio-macromolecular model drug encapsulated. In vitro physicochemical properties and in vitro drug release of FITC-BSA loaded SFNs (FITC-BSA-SFNs) were evaluated. Cytotoxicity, cellular uptake, and retention of FITC-BSA-SFNs were determined in human retinal pigment epithelial cell line (ARPE-19). In addition, in vivo distribution and safety of intravitreally administered FITC-BSA-SFNs were investigated in New Zealand white rabbits. The particle size of FITC-BSA-SFNs was 179.1 ± 3.7 nm with polydispersity index of 0.102 ± 0.033 and the zeta potential was greater than -25 mV. FITC-BSA-SFNs exhibited excellent biocompatibility with no cytotoxicity observed within 24 and 48 h in AREP-19 cells. Compared to FITC-BSA solution, FITC-BSA-SFNs showed enhanced cellular uptake and prolonged retention. Furthermore, FITC-BSA-SFNs achieved accumulated distribution and extended retention in retina in vivo following intravitreal injection compared to a single administration of free drug solution. Therefore, this bio-macromolecule delivery platform based on SFNs could have great potential in the treatment of posterior segment disorders.

Effects of different power multi-frequency ultrasound-assisted thawing on the quality characteristics and protein stability of large yellow croaker (Larimichthys crocea).

This study investigated the impact of multi-frequency ultrasound-assisted (20/28/40 kHz) thawing (MUAT) at different power levels (195, 220, 245, and 270 W, respectively) on the flesh quality and protein stability of large yellow croakers. Compared with flowing water thawing (FWT) and the other MUAT sample, flesh quality results indicated that the MUAT-220 W significantly reduced (p < 0.05) thawing loss, total volatile base nitrogen (TVB-N), total free amino acids (FAAs) and thiobarbituric acid reactive substances (TBARS). Low-field nuclear magnetic resonance (LF-NMR) spectroscopy indicated that MUAT-220 W samples had higher immobilized water content and lower free water content. In addition, the MUAT-220 W sample contained higher sulfhydryl and lower carbonyl contents compared to the FWT sample. Secondary and tertiary structural results of myofibrillar proteins (MPs) showed that MUAT-220 W significantly reduced thawing damage to MPs. Therefore, MUAT-220 W improved the quality and protein stability of the large yellow croaker during the defrosting process.

Effects of multi-frequency ultrasound-assisted immersion freezing processing on myofibrillar protein structure and lipid oxidation of large yellow croaker (Larimichthys crocea) during long-time frozen storage.

In this study, large yellow croaker (Larimichthys crocea) was frozen using multi-frequency ultrasound-assisted freezing (MUIF) with different powers (160 W, 175 W, and 190 W, respectively) and stored at -18 °C for ten months. The effect of different ultrasound powers on the myofibrillar protein (MP) structures and lipid oxidation of large yellow croaker was investigated. The results showed that MUIF significantly slowed down the oxidation of MP by inhibiting carbonyl formation and maintaining high sulfhydryl contents. These treatments also held a high activity of Ca2+-ATPase in the MP. MUIF maintained a higher ratio of α-helix to ß-sheet during frozen storage, thereby protecting the secondary structure of the tissue and stabilizing the tertiary structure. In addition, MUIF inhibited the production of thiobarbituric acid reactive substances value and the loss of unsaturated fatty acid content, indicating that MUIF could better inhibit lipid oxidation of large yellow croaker during long-time frozen storage.

The Endoplasmic Reticulum-Plasma Membrane Tethering Protein Ice2 Controls Lipid Droplet Size via the Regulation of Phosphatidylcholine in Candida albicans.

Lipid droplets (LDs) are intracellular organelles that play important roles in cellular lipid metabolism; they change their sizes and numbers in response to both intracellular and extracellular signals. Changes in LD size reflect lipid synthesis and degradation and affect many cellular activities, including energy supply and membrane synthesis. Here, we focused on the function of the endoplasmic reticulum-plasma membrane tethering protein Ice2 in LD dynamics in the fungal pathogen Candida albicans (C. albicans). Nile red staining and size quantification showed that the LD size increased in the ice2Δ/Δ mutant, indicating the critical role of Ice2 in the regulation of LD dynamics. A lipid content analysis further demonstrated that the mutant had lower phosphatidylcholine levels. As revealed with GFP labeling and fluorescence microscopy, the methyltransferase Cho2, which is involved in phosphatidylcholine synthesis, had poorer localization in the plasma membrane in the mutant than in the wild-type strain. Interestingly, the addition of the phosphatidylcholine precursor choline led to the recovery of normal-sized LDs in the mutant. These results indicated that Ice2 regulates LD size by controlling intracellular phosphatidylcholine levels and that endoplasmic reticulum-plasma membrane tethering proteins play a role in lipid metabolism regulation in C. albicans. This study provides significant findings for further investigation of the lipid metabolism in fungi.

Reduction of histone proteins dosages increases CFW sensitivity and attenuates virulence of Candida albicans.

Histone proteins are important components of nucleosomes, which play an important role in regulating the accessibility of DNA and the function of genomes. However, the effect of histone proteins dosages on physiological processes is not clear in the human fungal pathogen Candida albicans. In this study, we found that the deletion of the histone protein H3 coding gene HHT21 and the histone protein H4 coding gene HHF1 resulted in a significant decrease in the expression dosage of the histone proteins H3 and H4, which had a significant impact on the localization of the histone protein H2A and plasmid maintenance. Stress sensitivity experiments showed that the mutants hht21Δ/Δ, hhf1Δ/Δ and hht21Δ/Δhhf1Δ/Δ were more sensitive to cell wall stress induced by Calcofluor White (CFW) than the wild-type strain. Further studies showed that the decrease in the dosage of the histone proteins H3 and H4 led to the change of cell wall components, increased chitin contents, and down-regulated expression of the SAP9, KAR2, and CRH11 genes involved in the cell wall integrity (CWI) pathway. Overexpression of SAP9 could rescue the sensitivity of the mutants to CFW. Moreover, the decrease in the histone protein s dosages affected the FAD-catalyzed oxidation of Ero1 protein, resulting in the obstruction of protein folding in the ER, and thus reduced resistance to CFW. It was also found that CFW induced a large amount of ROS accumulation in the mutants, and the addition of ROS scavengers could restore the growth of the mutants under CFW treatment. In addition, the reduction of the histone proteins dosages greatly weakened systemic infection and kidney fungal burden in mice, and hyphal development was significantly impaired in the mutants under macrophage treatment, indicating that the histone proteins dosages is very important for the virulence of C. albicans. This study revealed that histone proteins dosages play a key role in the cell wall stress response and pathogenicity in C. albicans.

Effect of different stunning methods on antioxidant status, myofibrillar protein oxidation, and gelation properties of large yellow croaker during postmortem.

Post-mortem muscle biochemical processes play a crucial role on fish fillets quality and they are strictly linked to stunning methods. The improper stunning methods before slaughter could cause the fish to deteriorate more quickly during cold storage. This study aimed to investigate the effect of stunning methods (hit on the head, T1; gill cut, T2; immersion in ice/water slurry, T3; CO2 narcosis, T4; 40% CO2 + 30 % N2 + 30% O2, T5) on myofibrillar proteins (MPs) of large yellow croaker. The results indicated that T2 and T3 samples were significantly damaged compared with other samples, which reflected that the activities of total superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were significantly damaged during cold storage in T2 and T3 samples. And the gill cut and immersion in ice/water slurry resulted in the generation of protein carbonyl, the decrease of Ca2+-ATPase, free ammonia and protein solubility, and the production of dityrosine during storage. In addition, MPs gel of T2 and T3 samples showed the decrease of water hold capacity (WHC) and whiteness, structure destruction, and water migration. The T4 samples had the least damage of MPs and gel structure during cold storage.

Advances in biomimetic hydrogels for organoid culture.

An organoid is a 3-dimensional (3D) cell culture system that mimics the structural and functional characteristics of organs, and it has promising applications in regenerative medicine, precision drug screening and personalised therapy. However, current culture techniques of organoids usually use mouse tumour-derived scaffolds (Matrigel) or other animal-derived decellularised extracellular matrices as culture systems with poorly defined components and undefined chemical and physical properties, which limit the growth of organoids and the reproducibility of culture conditions. In contrast, some synthetic culture materials have emerged in recent years with well-defined compositions, and flexible adjustment and optimisation of physical and chemical properties, which can effectively support organoid growth and development and prolong survival time of organoid in vitro. In this review, we will introduce the challenge of animal-derived decellularised extracellular matrices in organoid culture, and summarise the categories of biomimetic hydrogels currently used for organoid culture, and then discuss the future opportunities and perspectives in the development of advanced hydrogels in organoids. We hope that this review can promote academic communication in the field of organoid research and provide some assistance in advancing the development of organoid cultivation technology.

Effects of Tricaine Methanesulphonate (MS-222) on Physiological Stress and Fresh Quality of Sea Bass (Lateolabrax maculatus) under Simulated High-Density and Long-Distance Transport Stress.

This study aimed to evaluate the effect of different transport densities on water deterioration, physiological response, nutrients, and fresh quality of sea bass (Lateolabrax maculatus) at 30 mg/L tricaine methanesulphonate (MS-222) before and after simulated live transport. The results indicated that the addition of MS-222 could effectively decrease mortality compared with the control (CK) sample during the simulated live transport. The concentration of dissolved oxygen was lower and the total ammonia nitrogen was higher in the high transport density samples than those of low transport density samples after 72 h in transport. The level of blood cortisol (COR), glucose (GLU), lactic acid (LD), aspartate aminotransferase (AST), alanine aminotransferase (ALT) for the sea bass were significantly higher compared with the CK sample (p < 0.05) during the simulated live transport and after 12 h of recovery. These results indicated that the sea bass presented a strong stress response in high-density transport. The glycogen, fat, and protein of the sea bass were degraded to supply the energy for the body in the process of surviving the transportation, resulting in the decreased nutrient content in the muscle, which recovered to the initial level (CK) after 12 h. The increase in flavor substance content, such as free amino acids, nucleotides, organic acids, and minerals, enhanced the special flavor of the muscle during the simulated live transport. This study demonstrates that the addition of MS-222 at 30 mg/L to the transport water is an effective method for live fish to realize low mortality and physiological response during high-density and long-distance transport.

Effect of different slaughter/stunning methods on stress response, quality indicators and susceptibility to oxidation of large yellow croaker (Larimichthys crocea).

This study aimed to investigate the effects of different slaughter methods (immersion in ice/water slurry, T1; gill cut, T2; CO2 asphyxia, T3; percussion (hit on the head with a stick), T4; Melissa officinalis L. essential oil + CO2, T5) on physiological stress, oxidative stress, and muscle quality in large yellow croaker. In terms of physiological stress, the levels of glucose (GLU), lactate dehydrogenase (LDH), and catalase (CAT) in CO2 asphyxia samples were significantly lower than those in other samples (p < 0.05). The level of cortisol (COR) in T1 sample was 1.25-1.84 times higher than that of other samples. The GLU level of T1 group was 3.2 times higher than that of T3 sample, and significantly higher than that of other samples. The creatine phosphokinase (CPK) and CAT levels of T2 samples were the highest (2.03 ng/mL and 8.34 U/mL, respectively). Furthermore, the superoxide dismutase (SOD) and glutathione peroxidase (GPx) analysis revealed that T3 and T4 samples could maintain good antioxidant enzyme activity during cold storage. The T3 samples maintained the stability of the protein (the lowest carbonyls and surface hydrophobicity) and reduced lipid oxidation (lower TBARS). In addition, the analysis of pH and water-holding capacity (WHC) revealed that T3 samples had better muscle quality. The muscle of T2 samples kept better color due to bloodletting treatment. The samples obtained after addition of Melissa officinalis L. essential oil had poorer indexes in all aspects compared to the T3 samples, which might be caused by the long anesthesia time of the essential oil.

Apoptotic Changes, Oxidative Stress and Immunomodulatory Effects in the Liver of Japanese Seabass (Lateolabrax japonicus) Induced by Ammonia-Nitrogen Stress during Keep-Live Transport.

This study investigated the effects of NH3-N on antioxidant responses, histoarchitecture, and immunity of Japanese seabass (Lateolabrax japonicus) during keep-live transport. The findings suggest that NH3-N stress transport alters the transcription of P53, Caspase 9, Bcl2, Caspase 3 and Bax genes, demonstrating that NH3-N stress can trigger the apoptotic pathway of P53-Bax-Bcl2 and Caspase and induce apoptosis. NH3-N stress transport also evoked transcriptional upregulation of inflammatory cytokines (tumor necrosis factor α (TNF-α), Toll-like receptor 3 (TLR-3), nuclear factor kappa ß (NF-κB), interleukin 6 (IL-6) and interleukin 1ß (IL-1ß)) and increased complement C3, C4, lysozyme (LZM) and immunoglobulin (IgM) levels, activating the innate immunological system during keep-live transport. In addition, NH3-N stress transport altered changes in the levels of superoxide dismutase (SOD), catalase (CAT), glutathione-related enzymes, and heat shock proteins 70 and 90 in the liver, indicating that the antioxidant system and Hsp protected the cells from NH3-N-induced oxidative stress. When excess ROS were not removed, they caused the body to respond with immunological and inflammatory responses, as well as apoptosis and tissue damage. This helps towards understanding the effect of NH3-N levels on sea bass during keep-live transport.

Transcriptional regulation of autophagy, cell wall stress response and pathogenicity by Pho23 in C. albicans.

The modification of chromatin by histone deacetylases (HDACs) has critical roles in transcriptional regulation. In this study, we identified the Rpd3 HDAC complex component Pho23 in Candida albicans and explored its role in the transcriptional regulation of physiological processes. PHO23 deletion increased autophagic activity and upregulated the transcription of ATG genes. Moreover, the deletion of PHO23 severely impaired cell wall stress resistance and reduced the cell wall integrity (CWI) pathway in response to cell wall stress. Furthermore, the pho23Δ/Δ mutant had partial defects in hyphal development and protease secretion, which were associated with the downregulation of genes involved in hyphal development (e.g. HWP1, ALS3 and ECE1) and genes encoding secreted aspartic proteases (e.g. SAP4, SAP5, SAP6 and SAP9). In addition, the deletion of PHO23 strongly attenuated systemic infection and kidney fungal burden in mice, demonstrating that Pho23 is required for the virulence of C. albicans. Together, our results revealed that Pho23 regulates many key physiological processes in C. albicans at the transcriptional level. These data also shed light on the potential for exploiting Rpd3 HDAC complex-related proteins as antifungal targets.

How we learn social norms: a three-stage model for social norm learning.

As social animals, humans are unique to make the world function well by developing, maintaining, and enforcing social norms. As a prerequisite among these norm-related processes, learning social norms can act as a basis that helps us quickly coordinate with others, which is beneficial to social inclusion when people enter into a new environment or experience certain sociocultural changes. Given the positive effects of learning social norms on social order and sociocultural adaptability in daily life, there is an urgent need to understand the underlying mechanisms of social norm learning. In this article, we review a set of works regarding social norms and highlight the specificity of social norm learning. We then propose an integrated model of social norm learning containing three stages, i.e., pre-learning, reinforcement learning, and internalization, map a potential brain network in processing social norm learning, and further discuss the potential influencing factors that modulate social norm learning. Finally, we outline a couple of future directions along this line, including theoretical (i.e., societal and individual differences in social norm learning), methodological (i.e., longitudinal research, experimental methods, neuroimaging studies), and practical issues.

Mec1-Rad53 Signaling Regulates DNA Damage-Induced Autophagy and Pathogenicity in Candida albicans.

DNA damage activates the DNA damage response and autophagy in C. albicans; however, the relationship between the DNA damage response and DNA damage-induced autophagy in C. albicans remains unclear. Mec1-Rad53 signaling is a critical pathway in the DNA damage response, but its role in DNA damage-induced autophagy and pathogenicity in C. albicans remains to be further explored. In this study, we compared the function of autophagy-related (Atg) proteins in DNA damage-induced autophagy and traditional macroautophagy and explored the role of Mec1-Rad53 signaling in regulating DNA damage-induced autophagy and pathogenicity. We found that core Atg proteins are required for these two types of autophagy, while the function of Atg17 is slightly different. Our results showed that Mec1-Rad53 signaling specifically regulates DNA damage-induced autophagy but has no effect on macroautophagy. The recruitment of Atg1 and Atg13 to phagophore assembly sites (PAS) was significantly inhibited in the mec1Δ/Δ and rad53Δ/Δ strains. The formation of autophagic bodies was obviously affected in the mec1Δ/Δ and rad53Δ/Δ strains. We found that DNA damage does not induce mitophagy and ER autophagy. We also identified two regulators of DNA damage-induced autophagy, Psp2 and Dcp2, which regulate DNA damage-induced autophagy by affecting the protein levels of Atg1, Atg13, Mec1, and Rad53. The deletion of Mec1 or Rad53 significantly reduces the ability of C. albicans to systematically infect mice and colonize the kidneys, and it makes C. albicans more susceptible to being killed by macrophages.

Atg8 and Ire1 in combination regulate the autophagy-related endoplasmic reticulum stress response in Candida albicans.

The unfolded protein response (UPR) is an important pathway to prevent endoplasmic reticulum (ER) stress in eukaryotic cells. In Saccharomyces cerevisiae, Ire1 is a key regulatory factor required for HAC1 gene splicing for further production of functional Hac1 and activation of UPR gene expression. Autophagy is another mechanism involved in the attenuation of ER stress by ER-phagy, and Atg8 is a core protein in autophagy. Both autophagy and UPR are critical for ER stress response, but whether they act individually or in combination in Candida albicans is unknown. In this study, we explored the interaction between Ire1 and the autophagy protein Atg8 for the ER stress response by constructing the atg8Δ/Δire1Δ/Δ double mutant in the pathogenic fungus C. albicans. Compared to the single mutants atg8Δ/Δ or ire1Δ/Δ, atg8Δ/Δire1Δ/Δ exhibited much higher sensitivity to various ER stress-inducing agents and more severe attenuation of UPR gene expression under ER stress. Further investigations showed that the double mutant had a defect in ER-phagy, which was associated with attenuated vacuolar fusion under ER stress. This study revealed that Ire1 and Atg8 in combination function in the activation of the UPR and ER-phagy to maintain ER homeostasis under ER stress in C. albicans.