Optogenetic induction of caspase-8 mediated apoptosis by employing Arabidopsis cryptochrome 2.

Apoptosis, a programmed cell death mechanism, is a regulatory process controlling cell proliferation as cells undergo demise. Caspase-8 serves as a pivotal apoptosis-inducing factor that initiates the death receptor-mediated apoptosis pathway. In this investigation, we have devised an optogenetic method to swiftly modulate caspase-8 activation in response to blue light. The cornerstone of our optogenetic tool relies on the PHR domain of Arabidopsis thaliana cryptochrome 2, which self-oligomerizes upon exposure to blue light. In this study, we have developed two optogenetic approaches for rapidly controlling caspase-8 activation in response to blue light in cellular systems. The first strategy, denoted as Opto-Casp8-V1, entails the fusion expression of the Arabidopsis blue light receptor CRY2 N-terminal PHR domain with caspase-8. The second strategy, referred to as Opto-Casp8-V2, involves the independent fusion expression of caspase-8 with the PHR domain and the CRY2 blue light-interacting protein CIB1 N-terminal CIB1N. Upon induction with blue light, PHR undergoes aggregation, leading to caspase-8 aggregation. Additionally, the blue light-dependent interaction between PHR and CIB1N also results in caspase-8 aggregation. We have validated these strategies in both HEK293T and HeLa cells. The findings reveal that both strategies are capable of inducing apoptosis, with Opto-Casp8-V2 demonstrating significantly superior efficiency compared to Opto-Casp8-V1.

The transcription factor SbHY5 mediates light to promote aluminum tolerance by activating SbMATE and SbSTOP1s expression.

Aluminum (Al) toxicity is a major factor limiting crop yields in acid soils. Sweet sorghum (Sorghum bicolor L.) is a high-efficient energy crop widely grown in tropical and subtropical regions of the world, where acid soil is common and Al toxicity is widespread. Here, we characterized a transcription factor SbHY5 in sweet sorghum, which mediated light to promote plant Al stress adaptation. The expression of SbHY5 was induced by Al stress and increasing light intensity. The overexpression of SbHY5 improved Al tolerance in transgenic plants, which was associated with increased citrate secretion and reduced Al content in roots. Meanwhile, SbHY5 was found to localize to the nucleus and displayed transcriptional activity. SbHY5 directly activated the expression of SbMATE, indicating that a HY5-MATE-dependent citrate secretion pathway is involved in Al tolerance in plants. SbSTOP1 was reported as a key transcription factor, regulating several Al tolerance genes. Here, inspiringly, we found that SbHY5 directly promoted the transcription of SbSTOP1, implying the existence of HY5-STOP1-Al tolerance genes-mediated regulatory pathways. Besides, SbHY5 positively regulated its own transcription. Our findings revealed a novel regulatory network in which a light signaling factor, SbHY5, confers Al tolerance in plants by modulating the expression of Al stress response genes.

Synthesis, Optical Properties and Cellular Toxicity of Water-Soluble near Infrared-II Fluorescent Assemblies Based on Pillar[5]arene.

The main challenges in second near-infrared region molecular fluorophores are poor water solubility and unknown long-term toxicity at present. Herein, new NIR-II molecular fluorophores have been designed and employed to integrate biocompatible pillar[5]arene with 10 outer triethylene oxide groups for the synthesis of rotaxane IRCR. In addition, PEGylated pillar[5]arenes have been combined for the self-assembly of two supramolecular vesicular systems, i.e., PP5-IR1 and PP5-IR2, affording aqueous solubility and lowered cellular toxicity. In aqueous solution, all these fluorophores displayed room-temperature emission with λmax at 986-1013 nm and quantum yields of 0.54-1.45%. They also exhibited good chemical stability and reasonable self-assembled sizes, which may find potential applications in NIR-II imaging. In addition, PP5-IR1 can be used as a fluorescent chemosensor for selective recognition of glutathione through the cleavage of dinitrophenyl ether and release the fluorescent dye.

Two Sweet Sorghum (Sorghum bicolor L.) WRKY Transcription Factors Promote Aluminum Tolerance via the Reduction in Callose Deposition.

Aluminum (Al) toxicity is a primary limiting factor for crop production in acidic soils. The WRKY transcription factors play important roles in regulating plant growth and stress resistance. In this study, we identified and characterized two WRKY transcription factors, SbWRKY22 and SbWRKY65, in sweet sorghum (Sorghum bicolor L.). Al induced the transcription of SbWRKY22 and SbWRKY65 in the root apices of sweet sorghum. These two WRKY proteins were localized in the nucleus and exhibited transcriptional activity. SbWRKY22 showed the significant transcriptional regulation of SbMATE, SbGlu1, SbSTAR1, SbSTAR2a, and SbSTAR2b, which are major known Al tolerance genes in sorghum. Interestingly, SbWRKY65 had almost no effect on the aforementioned genes, but it significantly regulated the transcription of SbWRKY22. Therefore, it is speculated that SbWRKY65 might indirectly regulate Al-tolerance genes mediated by SbWRKY22. The heterologous expression of SbWRKY22 and SbWRKY65 greatly improved the Al tolerance of transgenic plants. The enhanced Al tolerance phenotype of transgenic plants is associated with reduced callose deposition in their roots. These findings suggest the existence of SbWRKY22- and SbWRKY65-mediated Al tolerance regulation pathways in sweet sorghum. This study extends our understanding of the complex regulatory mechanisms of WRKY transcription factors in response to Al toxicity.

A novel tyrosine tRNA-derived fragment, tRFTyr, induces oncogenesis and lactate accumulation in LSCC by interacting with LDHA.

BACKGROUND: Transfer (t)RNA-derived small RNA (tsRNA), generated from precursor or mature tRNA, is a new type of small non-coding RNA (sncRNA) that has recently been shown to play a vital role in human cancers. However, its role in laryngeal squamous cell carcinoma (LSCC) remains unclear. METHODS: We elucidated the expression profiles of tsRNAs in four paired LSCC and non-neoplastic tissues by sequencing and verified the sequencing data by quantitative real-time PCR (qRT-PCR) of 60 paired samples. The tyrosine-tRNA derivative tRFTyr was identified as a novel oncogene in LSCC for further study. Loss-of-function experiments were performed to evaluate the roles of tRFTyr in tumorigenesis of LSCC. Mechanistic experiments including RNA pull-down, parallel reaction monitoring (PRM) and RNA immunoprecipitation (RIP) were employed to uncover the regulatory mechanism of tRFTyr in LSCC. RESULTS: tRFTyr was significantly upregulated in LSCC samples. Functional assays showed that knockdown of tRFTyr significantly suppressed the progression of LSCC. A series of mechanistic studies revealed that tRFTyr could enhance the phosphorylated level of lactate dehydrogenase A (LDHA) by interacting with it. The activity of LDHA was also activated, which induced lactate accumulation in LSCC cells. CONCLUSIONS: Our data delineated the landscape of tsRNAs in LSCC and identified the oncogenic role of tRFTyr in LSCC. tRFTyr could promote lactate accumulation and tumour progression in LSCC by binding to LDHA. These findings may aid in the development of new diagnostic biomarkers and provide new insights into therapeutic strategies for LSCC.

Comparison of the clinical features of HIV-positive and HIV-negative hosts infected with Talaromyces marneffei: A multicenter, retrospective study.

OBJECTIVES: Talaromyces marneffei is an emerging pathogen, and the number of infections in HIV-negative individuals is rapidly increasing. Nevertheless, there is no sufficient comprehensive report on this issue, and awareness needs to be raised among clinicians. METHODS: We analyzed the differences in the clinical data of patients who are HIV-negative and HIV-positive with Talaromyces marneffei infection (TMI) from 2018 to 2022. RESULTS: A total of 848 patients were included, among whom 104 were HIV-negative. The obvious differences between the HIV-positive and HIV-negative groups were as follows: (i) the patients who are HIV-negative were older and more likely to exhibit cough and rash, (ii) the time in days from symptom onset to diagnosis among patients who are HIV-negative was longer, (iii) the laboratory findings and radiological presentations seemed more severe in patients who are HIV-negative, (iv) differences were observed regarding the underlying conditions and co-infection pathogens, and correlation analysis showed that correlations existed for many indicators, (v) and persistent infection was more likely to occur in patients who are HIV-negative. CONCLUSION: TMI in patients who are HIV-negative differs from that in patients who are HIV-positive in many aspects, and more investigations are needed. Clinicians should be more aware of TMI in patients who are HIV-negative.

Chidamide, a novel histone deacetylase inhibitor, inhibits laryngeal cancer progression in vitro and in vivo.

Although surgery is an important treatment for laryngeal cancer, surgery has a significant negative impact on the quality of life of patients, and many patients have poor tolerance to surgery. Therefore, alternative chemotherapeutic drugs are an important research hotspot. Chidamide is a histone deacetylase inhibitor that selectively inhibits the expression of type I and IIb histone deacetylases (1, 2, 3 and 10). It has a significant anticancer effect on a variety of solid tumours. This study verified the inhibitory effect of chidamide on laryngeal carcinoma. We conducted a variety of cellular and animal experiments to explore how chidamide inhibits the development of laryngeal cancer. The results showed that chidamide had significant antitumour activity against laryngeal carcinoma cells and xenografts and could induce cell apoptosis, ferroptosis and pyroptosis. This study provides a potential option for the treatment of laryngeal cancer.

Characterization of GmMATE13 in its contribution of citrate efflux and aluminum resistance in soybeans.

Citrate exudation mediated by a citrate transporter of the MATE protein family is critical for resisting aluminum (Al) toxicity in soybeans. However, the expression patterns of citrate transporter genes differ under Al stress. Thus, exploring the responsive pattern of GmMATEs in response to Al stress is of great importance to understand the Al resistance mechanism in soybeans. In the present study, the phylogenetic analysis, transcriptionally expressed pattern, and function of GmMATE13 were investigated. The results show that soybean GmMATE13 is highly homologous to known citrate transporter proteins from other plants. Under Al exposure, the transcript abundance of GmMATE13 was increased during a 24 h Al treatment period. The expression of GmMATE13 is specifically induced by Al exposure, but not by the status of Fe, Cu, Cd, or La. Moreover, it was also highly increased when soybean seedlings were grown on acidic soil with a high Al content. Subcellular localization showed that GmMATE13 was localized on the plasma membrane when it was transiently expressed in Arabidopsis protoplasts. Investigation of tissue localization of GmMATE13 expression by investigating GUS activity staining under control of the GmMATE13 promoter showed that it was mainly expressed in the central cylinder in the root tips of the soybean under Al-free conditions, yet extended to cortical and epidermis cells under Al stress. Finally, overexpressing GmMATE13 in soybean hairy roots enhanced Al resistance by increasing citrate efflux. Collectively, we conclude that GmMATE13 is a promising candidate to improve the resistance of soybean to Al toxicity in acidic soil.

Effect of Tool Coatings on Machining Properties of Compacted Graphite Iron.

Compacted graphite iron (CGI) has become the most ideal material for automotive engine manufacturing owing to its excellent mechanical properties. However, tools are severely worn during processing, considerably shortening their lifespan. In this study, we prepared a series of cemented carbide-coated tools and evaluated their coating properties in cutting tests. Among all tested coatings, PVD coating made of AlCrN (AC) presented with the best surface integrity and mechanical properties, achieving the best comprehensive performance in the coating test. The AC-coated tool also exhibited the best cutting performance at a low speed of 120 m/min, corresponding to a 60% longer cutting life and the lowest workpiece surface roughness relative to other coated tools. In the cutting test at a high speed of 350 m/min, the CVD double-layer coated tool (MT) with a TiCN inner layer of and an Al2O3 outer layer had a 70% longer cutting life and the lowest workpiece surface roughness relative to other coated tools.

Development and regeneration of periodontal supporting tissues.

Periodontal tissues, including gingiva, cementum, periodontal ligament, and alveolar bone, play important roles in oral health. Under physiological conditions, periodontal tissues surround and support the teeth, maintaining the stability of the teeth and distributing the chewing forces. However, under pathological conditions, with the actions of various pathogenic factors, the periodontal tissues gradually undergo some irreversible changes, that is, gingival recession, periodontal ligament rupture, periodontal pocket formation, alveolar bone resorption, eventually leading to the loosening and even loss of the teeth. Currently, the regenerations of the periodontal tissues are still challenging. Therefore, it is necessary to study the development of the periodontal tissues, the principles and processes of which can be used to develop new strategies for the regeneration of periodontal tissues. This review summarizes the development of periodontal tissues and current strategies for periodontal healing and regeneration.

Electrochemical Mechanism of Molten Salt Electrolysis from TiO2 to Titanium.

Electrochemical mechanisms of molten salt electrolysis from TiO2 to titanium were investigated by Potentiostatic electrolysis, cyclic voltammetry, and square wave voltammetry in NaCl-CaCl2 at 800 °C. The composition and morphology of the product obtained at different electrolysis times were characterized by XRD and SEM. CaTiO3 phase was found in the TiO2 electrochemical reduction process. Electrochemical reduction of TiO2 to titanium is a four-step reduction process, which can be summarized as TiO2→Ti4O7→Ti2O3→TiO→Ti. Spontaneous and electrochemical reactions take place simultaneously in the reduction process. The electrochemical reduction of TiO2→Ti4O7→Ti2O3→TiO affected by diffusion was irreversible.

Arabidopsis cryptochrome 2 forms photobodies with TCP22 under blue light and regulates the circadian clock.

Cryptochromes are blue light receptors that regulate plant growth and development. They also act as the core components of the central clock oscillator in animals. Although plant cryptochromes have been reported to regulate the circadian clock in blue light, how they do so is unclear. Here we show that Arabidopsis cryptochrome 2 (CRY2) forms photobodies with the TCP22 transcription factor in response to blue light in plant cells. We provide evidence that PPK kinases influence the characteristics of these photobodies and that together these components, along with LWD transcriptional regulators, can positively regulate the expression of CCA1 encoding a central component of the circadian oscillator.

Talaromyces marneffei and Mycobacterium tuberculosis co-infection in a patient with high titer anti-interferon-γ autoantibodies: a case report.

BACKGROUND: High-titer anti-interferon (IFN)-γ autoantibodies are strongly associated with intracellular pathogens such as nontuberculous mycobacteria and Talaromyces marneffei, but they are not as commonly associated with Talaromyces marneffei co-infected with Mycobacterium tuberculosis. CASE PRESENTATION: Herein, we report a case of an HIV-negative Chinese man with a severe, disseminated co-infection of Talaromyces marneffei and Mycobacterium tuberculosis, who had a high-titer of anti IFN-γ autoantibodies and a CFI heterozygous nonsense gene mutation. The patient rapidly developed sepsis and died. Through by flow cytometry for CD4+ T cells' intracellular phosphorylated STAT-1 and Th1 cells (CD4+ IFN-γ+ cells), we found that the patient's serum can inhibited IFN γ-induced CD4+ T cells' STAT-1 phosphorylation and Th1 cell differentiation in normal peripheral blood mononuclear cells, but this phenomenon was not observed in normal control's serum. In addition, the higher serum concentration in the culture medium, the more obvious inhibition of Th1 cell differentiation. CONCLUSIONS: For HIV-negative individuals with relapsing, refractory, fatal double or multiple intracellular pathogen infections, especially Talaromyces marneffei, clinicians should be aware that if they might be dealing with adult-onset immunodeficiency syndrome due to high-titer anti-IFN-γ autoantibodies. Systematic genetic and immunological investigations should also be performed.

Multifunctional 3D printed porous GelMA/xanthan gum based dressing with biofilm control and wound healing activity.

Bacterial infections are the major challenges of wound treatment in current clinical applications. In this study, Three-dimensional (3D) antibacterial wound dressing has been fabricated via introducing N-halamine/TiO2 to gelatin methacrylate and xanthan gum. The prepared 3D printed dressings showed ideal swelling ratio and excellent water uptake efficiency. TiO2 nanoparticles were introduced by in-situ to improve the ultraviolet stability of N-halamines. The 3D printed GX2-TiO2-PSPH-Cl prepared dressings containing titanium dioxide retained 0.19% active chlorine after ultraviolet irradiation for 20 min, which was much higher than that of N-halamine dressings without the addition of TiO2. The 3D printed dressings showed good antibacterial activity, and 100% of Escherichia coli O157:H7 and Staphylococcus aureus were inactivated after 60 min of contact. Furthermore, the biofilm test indicated that the 3D antibacterial dressings were able to inhibit the formation of bacterial biofilm. The 3D printed dressings possess outstanding biocompatibility. Moreover, in vivo data demonstrated that the 3D printed dressings could significantly accelerate wound healing in a mouse model, indicating that the developed 3D printed dressings are ideal candidates for wound treatment.

Roles of a Cryptochrome in Carbon Fixation and Sucrose Metabolism in the Liverwort Marchantia polymorpha.

In vascular plants, cryptochromes acting as blue-light photoreceptors have various functions to adapt plants to the fluctuating light conditions on land, while the roles of cryptochromes in bryophytes have been rarely reported. In this study, we investigated functions of a single-copy ortholog of cryptochrome (MpCRY) in the liverwort Marchantia polymorpha. Knock-out of MpCRY showed that a large number of the mutant plants exhibited asymmetric growth of thalli under blue light. Transcriptome analyses indicated that MpCRY is mainly involved in photosynthesis and sugar metabolism. Further physiological analysis showed that Mpcry mutant exhibited a reduction in CO2 uptake and sucrose metabolism. In addition, exogenous application of sucrose or glucose partially restored the symmetrical growth of the Mpcry mutant thalli. Together, these results suggest that MpCRY is involved in the symmetrical growth of thallus and the regulation of carbon fixation and sucrose metabolism in M. polymorpha.

Talaromyces Marneffei Infection in Lung Cancer Patients with Positive AIGAs: A Rare Case Report.

BACKGROUND: Talaromyces marneffei is considered to commonly cause infection in individuals with human immunodeficiency virus (HIV) infection. However, the epidemiology of T. marneffei has changed, and an increasing number of HIV-negative but immunodeficient patients are infected with T. marneffei. The mechanisms of T. marneffei infection of HIV-negative hosts are complex and diverse. We report 2 cases of HIV-negative lung cancer with T. marneffei infection and positive anti-interferon-gamma autoantibodies (AIGAs) to provide clinical experience. CASE PRESENTATION: We report lung adenocarcinoma combined with T. marneffei infection in HIV-negative patients, and their AIGAs were measured. Both patients were male with a family history of cancer and presented with recurrent fever and cough. The patients were negative for HIV antibodies but positive for AIGAs. Chest computed tomography (CT) showed pulmonary nodules, exudative lesions and solid changes. The patients were diagnosed with lung adenocarcinoma and Talaromycosis marneffei (TSM) by pathological examination and tissue culture. Patient 1 received only antifungal treatment, refused antitumor treatment and died in February 2019, and Patient 2 unfortunately died in April 2019 after antifungal and antitumor treatments. CONCLUSION: An increasing number of HIV-negative but immunodeficient patients are infected with T. marneffei. The 2 patients in this report had lung cancer and positive AIGAs, causing immunodeficiencies, but the mechanism of T. marneffei infection in such patients is complex. Clinically, we should consider a comprehensive immunological examination to avoid the omission of other immunodeficiencies. We recommend routine testing for AIGA levels in HIV-negative marneffei patients. It is difficult to distinguish between lung cancer and disseminated TSM due to similar clinical characteristics and imaging, and multiple biopsies and cultures of diseased tissue are necessary. Early antifungal treatment and standard antitumor treatment can achieve satisfactory curative effects when a patient has both diseases.

The Arabidopsis HY2 Gene Acts as a Positive Regulator of NaCl Signaling during Seed Germination.

Phytochromobilin (PΦB) participates in the regulation of plant growth and development as an important synthetase of photoreceptor phytochromes (phy). In addition, Arabidopsis long hypocotyl 2 (HY2) appropriately works as a key PΦB synthetase. However, whether HY2 takes part in the plant stress response signal network remains unknown. Here, we described the function of HY2 in NaCl signaling. The hy2 mutant was NaCl-insensitive, whereas HY2-overexpressing lines showed NaCl-hypersensitive phenotypes during seed germination. The exogenous NaCl induced the transcription and the protein level of HY2, which positively mediated the expression of downstream stress-related genes of RD29A, RD29B, and DREB2A. Further quantitative proteomics showed the patterns of 7391 proteins under salt stress. HY2 was then found to specifically mediate 215 differentially regulated proteins (DRPs), which, according to GO enrichment analysis, were mainly involved in ion homeostasis, flavonoid biosynthetic and metabolic pathways, hormone response (SA, JA, ABA, ethylene), the reactive oxygen species (ROS) metabolic pathway, photosynthesis, and detoxification pathways to respond to salt stress. More importantly, ANNAT1-ANNAT2-ANNAT3-ANNAT4 and GSTU19-GSTF10-RPL5A-RPL5B-AT2G32060, two protein interaction networks specifically regulated by HY2, jointly participated in the salt stress response. These results direct the pathway of HY2 participating in salt stress, and provide new insights for the plant to resist salt stress.

Identification of a bacterial-type ATP-binding cassette transporter implicated in aluminum tolerance in sweet sorghum (Sorghum bicolor L.).

Aluminum (Al) toxicity in acidic soils severely reduces crop production worldwide. Sorghum (Sorghum bicolor L.) is an important agricultural crop widely grown in tropical and subtropical regions, where Al toxicity is prevalent. ATP-binding cassette (ABC) transporters play key roles in the development of plants and include the member sensitive to aluminum rhizotoxicity 1 (STAR1), which is reported to be associated with Al tolerance in a few plant species. However, a STAR1 homolog has not been characterized in sorghum with respect to Al tolerance. Here, we identified and characterized a SbSTAR1 gene in sweet sorghum encoding the nucleotide-binding domain of a bacterial-type ABC transporter. The transcriptional expression of SbSTAR1 is induced by Al in a time- and dosage-dependent manner in root, especially in root tip, which is the key site of Al toxicity in plants. The typical Al-associated transcription factor SbSTOP1 showed transcriptional regulation of SbSTAR1. SbSTAR1 was present at both the cytoplasm and nuclei. Overexpression of SbSTAR1 significantly enhanced the Al tolerance of transgenic plants, which possibly via regulating the hemicellulose content in root cell wall. This study provides the first ABC protein in sorghum implicated in Al tolerance, suggesting the existence of a SbSTAR1-mediated Al tolerance mechanism in sorghum.

Arabidopsis CIB3 regulates photoperiodic flowering in an FKF1-dependent way.

Arabidopsis cryptochrome 2 (CRY2) and FLAVIN-BINDING, KELCH REPEAT, and F-BOX 1 (FKF1) are blue light receptors mediating light regulation of growth and development, such as photoperiodic flowering. CRY2 interacts with a basic helix-loop-helix transcription factor CIB1 in response to blue light to activate the transcription of the flowering integrator gene FLOWERING LOCUS T (FT). CIB1, CIB2, CIB4, and CIB5 function redundantly to promote flowering in a CRY2-dependent way and form various heterodimers to bind to the noncanonical E-box sequence in the FT promoter. However, the function of CIB3 has not been described. We discovered that CIB3 promotes photoperiodic flowering independently of CRY2. Moreover, CIB3 does not interact with CRY2 but interacts with CIB1 and functions synergistically with CIB1 to promote the transcription of the GI gene. FKF1 is required for CIB3 to promote flowering and enhances the CIB1-CIB3 interaction in response to blue light.

Two new species of Helochares, with additional faunistic records from China (Coleoptera, Hydrophilidae, Acidocerinae).

Two new species, Helocharesguoi Yang & Jia, sp. nov. and Helocharesdistinctus Jia & Tang, sp. nov., are described. Two species are recorded for the first time from China: Helocharesnegatus Hebauer, 1995 from Yunnan, and Helocharesminusculus d'Orchymont, 1943 from Guangdong. Additional faunistic data from China are provided for the following species: Helochareshainanensis Dong & Bian, 2021, Helocharesnipponicus Hebauer, 1995, Helocharessauteri d'Orchymont, 1943, Helocharesdensus Sharp, 1890, Helochareslentus Sharp, 1890, Helocharesneglectus (Hope, 1854) and Helocharesanchoralis Sharp, 1890. The Chinese fauna of Helochares comprises 16 species, 11 of which are illustrated in this contribution. Helocharescrenatus Régimbart, 1921 is removed from the Chinese fauna.