Integrated chromosomal instability and tumor microbiome redefined prognosis-related subtypes of pancreatic cancer.

BACKGROUND: Pancreatic cancer is a common malignancy with poor prognosis and limited treatment. Here we aimed to investigate the role of host chromosomal instability (CIN) and tumor microbiome in the prognosis of pancreatic cancer patients. METHODS: One hundred formalin-fixed paraffin-embedded (FFPE) pancreatic cancer samples were collected. DNA extracted from FFPE samples were analyzed by low-coverage whole-genome sequencing (WGS) via a customized bioinformatics workflow named ultrasensitive chromosomal aneuploidy detector. RESULTS: Samples are tested according to the procedure of ultrasensitive chromosomal aneuploidy detector (UCAD). We excluded 2 samples with failed quality control, 1 patient lost to follow-up and 6 dead in the perioperative period. The final 91 patients were admitted for the following analyses. Thirteen (14.3%) patients with higher CIN score had worse overall survival (OS) than those with lower CIN score. The top 20 microbes in pancreatic cancer samples included 15 species of bacteria and 5 species of viruses. Patients with high human herpesvirus (HHV)-7 and HHV-5 DNA reads exhibited worse OS. Furthermore, we classified 91 patients into 3 subtypes. Patients with higher CIN score (n =13) had the worst prognosis (median OS 6.9 mon); patients with lower CIN score but with HHV-7/5 DNA load (n = 24) had worse prognosis (median OS 10.6 mon); while patients with lower CIN score and HHV-7/5 DNA negative (n = 54) had the best prognosis (median OS 21.1 mon). CONCLUSIONS: High CIN and HHV-7/5 DNA load were associated with worse survival of pancreatic cancer. The novel molecular subtypes of pancreatic cancer based on CIN and microbiome had prognostic value.

An integrated proteomics and metabolomics approach to assess graft quality and predict early allograft dysfunction after liver transplantation: a retrospective cohort study.

BACKGROUND: Early allograft dysfunction (EAD) is a common complication after liver transplantation (LT) and is associated with poor prognosis. Graft itself plays a major role in the development of EAD. We aimed to reveal the EAD-specific molecular profiles to assess graft quality and establish EAD predictive models. METHODS: A total of 223 patients who underwent LT were enrolled and divided into training (n=73) and validation (n=150) sets. In the training set, proteomics was performed on graft biopsies, together with metabolomics on paired perfusates. Differential expression, enrichment analysis, and protein-protein interaction network were used to identify the key molecules and pathways involved. EAD predictive models were constructed using machine learning and verified in the validation set. RESULTS: A total of 335 proteins were differentially expressed between the EAD and non-EAD groups. These proteins were significantly enriched in triglyceride and glycerophospholipid metabolism, neutrophil degranulation, and the MET-related signaling pathway. The top 12 graft proteins involved in the aforementioned processes were identified, including GPAT1, LPIN3, TGFB1, CD59, and SOS1. Moreover, downstream metabolic products, such as lactate dehydrogenase, interleukin-8, triglycerides, and the phosphatidylcholine/phosphorylethanolamine ratio in the paired perfusate displayed a close relationship with the graft proteins. To predict the occurrence of EAD, an integrated model using perfusate metabolic products and clinical parameters showed areas under the curve of 0.915 and 0.833 for the training and validation sets, respectively. It displayed superior predictive efficacy than that of currently existing models, including donor risk index and D-MELD scores. CONCLUSIONS: We identified novel biomarkers in both grafts and perfusates that could be used to assess graft quality and provide new insights into the etiology of EAD. Herein, we also offer a valid tool for the early prediction of EAD.

Regio- and Enantioselective Construction of Tetrazole Hemiaminal Esters and Related Prodrugs via Biocatalytic Dynamic Kinetic Resolution.

Enzyme-catalyzed dynamic kinetic resolution was applied to the one-pot regio- and enantioselective synthesis of 2,5-disubstituted tetrazole hemiaminal esters, among which 72% of the products were obtained in excellent enantiopurities (99% ees). Tunable stereoselectivity was achieved by using different types of enzymes during the synthesis of a key intermediate for a clinic drug candidate. Successful preparation of tetrazole ester prodrugs and high catalyst recyclability further demonstrated the potential practical application of this protocol.

Changes in CO2 concentration drive a succession of toxic and non-toxic strains of Microcystis blooms.

The dynamic changes between toxic and non-toxic strains of Microcystis blooms have always been a hot topic. Previous studies have found that low CO2 favors toxic strains, but how changing dissolved CO2 (CO2 [aq]) in water body influences the succession of toxic and non-toxic strains in Microcystis blooms remains uncertain. Here, we combined laboratory competition experiments, field observations, and a machine learning model to reveal the links between CO2 changes and the succession. Laboratory experiments showed that under low CO2 conditions (100-150 ppm), the toxic strains could make better use of CO2 (aq) and be dominant. The non-toxic strains demonstrated a growth advantage as CO2 concentration increased (400-1000 ppm). Field observations from June to November in Lake Taihu showed that the percentage of toxic strains increased as CO2 (aq) decreased. Machine learning highlighted links between the inorganic carbon concentration and the proportion of advantageous strains. Our findings provide new insights for cyanoHABs prediction and prevention.

Scoparone from Artemisia capillaris Thunb. induces apoptosis in HepG2 cells via activation of both intracellular and extracellular pathways.

Six separated compounds were identified from Artemisia capillaris Thunb., and they were 7-methoxycoumarin (1), 6,7-dimethoxycoumarin (2), 7-hydroxy-6-methoxycoumarin (3), quercetin (4), chlorogenic acid (5) and caffeic acid (6). Among them, 6,7-dimethoxycoumarin, as known as scoparone, was the most effective on scavenging ABTS free radicals (IC50 = 0.97 µΜ) and was then tested by cytotoxic activity and pro-apoptotic activity against HepG2 cells. Scoparone dose-dependently and time-dependently inhibited the cell proliferation. Furthermore, scoparone induced the expression of Bax, concurrently suppressing the expression of Bcl-2, resulting in a noteworthy elevation in the Bax/Bcl-2 ratio to up-regulate Caspase-3 activity, thus inducing cell apoptosis via the intracellular pathway. Meanwhile, scoparone promoted the expression of Fas, FasL, FADD, Caspase-8 and Caspase-3, indicating that scoparone also triggered apoptosis via the extracellular pathway. In a word, scoparone demonstrated remarkable antitumor capability to induce apoptosis of HepG2 cells through both intracellular and extracellular pathways.

Combination of ultrasound and serological tests for detecting occult lateral lymph node metastases in medullary thyroid cancer.

PURPOSE: To investigate the value of ultrasound and serum marker tests in detecting lateral lymph node metastasis in medullary thyroid cancer (MTC). METHODS: Data of 105 patients diagnosed with MTC and admitted to the Department of General Surgery in Peking Union Medical College Hospital from June 2010 to August 2020 were collected and retrospectively analyzed. RESULTS: Ultrasound examination alone had a sensitivity of 89.36% and a specificity of 70.69%. For surveillance of postoperative carcinoembryonic antigen and calcitonin, cut-off values of 7.115 ng/mL and 13.185 pg/mL, respectively, were shown to discriminate the presence of cervical lymph node metastasis. Combining ultrasound and postoperative serum levels of both carcinoembryonic antigen and calcitonin as serial tests increased the specificity to 91.38% and 87.93%, with a sensitivity of 95.45%. Multivariate logistic analysis identified the following risk factors for lateral lymph node metastasis in MTC: suspicious lymph nodes detected by ultrasound and postoperative calcitonin above 13.185 pg/mL. CONCLUSION: The combination of ultrasound and serological tests achieved higher sensitivity and specificity to identify MTC cases with potential occult lateral cervical lymph node metastasis compared with single tests.

Immunotherapy for hepatocellular carcinoma recurrence after liver transplantation, can we harness the power of immune checkpoint inhibitors?

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related death globally and liver transplantation (LT) can serve as the best curative treatment option. However, HCC recurrence after LT remains the major obstacle to the long-term survival of recipients. Recently, immune checkpoint inhibitors (ICIs) have revolutionized the treatment of many cancers and provided a new treatment strategy for post-LT HCC recurrence. Evidence has been accumulated with the real-world application of ICIs in patients with post-LT HCC recurrence. Notably, the use of these agents as immunity boosters in recipients treated with immunosuppressors is still controversial. In this review, we summarized the immunotherapy for post-LT HCC recurrence and conducted an efficacy and safety evaluation based on the current experience of ICIs for post-LT HCC recurrence. In addition, we further discussed the potential mechanism of ICIs and immunosuppressive agents in regulating the balance between immune immunosuppression and lasting anti-tumor immunity.

Decoding Single-cell Landscape and Intercellular Crosstalk in the Transplanted Liver.

BACKGROUND: Liver transplantation (LT) is the most effective treatment for various end-stage liver diseases. However, the cellular complexity and intercellular crosstalk of the transplanted liver have constrained analyses of graft reconstruction after LT. METHODS: We established an immune-tolerated orthotopic LT mouse model to understand the physiological process of graft recovery and intercellular crosstalk. We employed single-cell RNA sequencing and cytometry by time-of-flight to comprehensively reveal the cellular landscape. RESULTS: We identified an acute and stable phase during perioperative graft recovery. Using single-cell technology, we made detailed annotations of the cellular landscape of the transplanted liver and determined dynamic modifications of these cells during LT. We found that 96% of graft-derived immune cells were replaced by recipient-derived cells from the preoperative to the stable phase. However, CD206 + MerTK + macrophages and CD49a + CD49b - natural killer cells were composed of both graft and recipient sources even in the stable phase. Intriguingly, the transcriptional profiles of these populations exhibited tissue-resident characteristics, suggesting that recipient-derived macrophages and natural killer cells have the potential to differentiate into 'tissue-resident cells' after LT. Furthermore, we described the transcriptional characteristics of these populations and implicated their role in regulating the metabolic and immune remodeling of the transplanted liver. CONCLUSIONS: In summary, this study delineated a cell atlas (type-proportion-source-time) of the transplanted liver and shed light on the physiological process of graft reconstruction and graft-recipient crosstalk.

Spring Viremia of Carp Virus N Protein Negatively Regulates IFN Induction through Autophagy-Lysosome-Dependent Degradation of STING.

Fish possess a powerful IFN system to defend against aquatic virus infections. Nevertheless, spring viremia of carp virus (SVCV) causes large-scale mortality in common carp and significant economic losses to aquaculture. Therefore, it is necessary to investigate the strategies used by SVCV to escape the IFN response. In this study, we show that the SVCV nucleoprotein (N protein) negatively regulates cellular IFN production by degrading stimulator of IFN genes (STING) via the autophagy-lysosome-dependent pathway. First, overexpression of N protein inhibited the IFN promoter activation induced by polyinosinic-polycytidylic acid and STING. Second, the N protein associated with STING and experiments using a dominant-negative STING mutant demonstrated that the N-terminal transmembrane domains of STING were indispensable for this interaction. Then, the N protein degraded STING in a dose-dependent and autophagy-lysosome-dependent manner. Intriguingly, in the absence of STING, individual N proteins could not elicit host autophagic flow. Furthermore, the autophagy factor Beclin1 was found to interact with the N protein to attenuate N protein-mediated STING degradation after beclin1 knockdown. Finally, the N protein remarkably weakened STING-enhanced cellular antiviral responses. These findings reveal that SVCV uses the host autophagic process to achieve immune escape, thus broadening our understanding of aquatic virus pathogenesis.

Evolution of Composition and Methane Occurrence of Bituminous Coal after Igneous Intrusion: A Case Study of Daxing Coal Mine, Tiefa Basin, China.

Since 1987, there have been 10 coal and gas outbursts in Daxing Coal Mine, Tiefa Basin. It was reported that these outbursts were related to igneous intrusion. To study the influence of igneous intrusion into bituminous on coal bed methane (CBM) occurrence, eight coal samples were collected at different distances (0.2-10 m) from an intrusion in Daxing Coal Mine. The petrographic and chemical data, pore characteristics, and adsorption properties of unaltered and heat-affected coals were tested and compared. Approaching the dike, vitrinite reflectance (R o) increases from 0.56 to 1.14%, while moisture decreases from 6.4 to 3.1%. According to R o, with the thermal evolution of igneous rocks, the temperature of heat-affected coal seams generally increases from ∼78 to ∼169 °C. The bituminous coal contained high moisture content before the intrusion, and after the igneous intrusion, partial heat may be used for moisture gasification; the other heat was used to improve coal metamorphism, resulting in little increase in the coal metamorphism as expected. The synergistic effect of igneous thermal evolution and moisture content change improves the adsorption capacity of coal. Affected by the igneous intrusion, the gas content of the heat-affected coal seams No. 4 (above the sill) and No. 7 (below the sill) is higher than that of the unaltered No. 12 coal seam, and the gas content of No. 7 coal seam (beneath the sill) is higher than that of No. 4 coal seam. The low permeability sill has a sealing effect on the gas in No. 7 coal seam. Therefore, the evolution process of CBM occurrence in the Daxing Coal Mine can be speculated, that is, after magma intrusion, the temperature increased, the metamorphic degree of coal increased, and water gasification occurred at the same time. The original adsorption site occupied by moisture was vacated, which improved the methane adsorption capacity of the coal and provided favorable conditions for CBM occurrence.

Synergistic Hypolipidemic Effects and Mechanisms of Phytochemicals: A Review.

Hyperlipidemia, a chronic disorder of abnormal lipid metabolism, can induce obesity, diabetes, and cardiovascular and cerebrovascular diseases such as coronary heart disease, atherosclerosis, and hypertension. Increasing evidence indicates that phytochemicals may serve as a promising strategy for the prevention and management of hyperlipidemia and its complications. At the same time, the concept of synergistic hypolipidemic and its application in the food industry is rapidly increasing as a practical approach to preserve and improve the health-promoting effects of functional ingredients. The current review focuses on the effects of single phytochemicals on hyperlipidemia and its mechanisms. Due to the complexity of the lipid metabolism regulatory network, the synergistic regulation of different metabolic pathways or targets may be more effective than single pathways or targets in the treatment of hyperlipidemia. This review summarizes for the first time the synergistic hypolipidemic effects of different combinations of phytochemicals such as combinations of the same category of phytochemicals and combinations of different categories of phytochemicals. In addition, based on the different metabolic pathways or targets involved in synergistic effects, the possible mechanisms of synergistic hypolipidemic effects of the phytochemical combination are illustrated in this review. Hence, this review provides clues to boost more phytochemical synergistic hypolipidemic research and provides a theoretical basis for the development of phytochemicals with synergistic effects on hyperlipidemia and its complications.

The role of macrophage TAM receptor family in the acute-to-chronic progression of liver disease: From friend to foe?

Hepatic macrophages, the key cellular components of the liver, emerge as essential players in liver inflammation, tissue repair and subsequent fibrosis, as well as tumorigenesis. Recently, the TAM receptor tyrosine kinase family, consisting of Tyro3, Axl and MerTK, was found to be a pivotal modulator of macrophages. Activation of macrophage TAM receptor signalling promotes the efferocytosis of apoptotic cells and skews the polarization of macrophages. After briefly reviewing the mechanisms of TAM receptor signalling in macrophage polarization, we focus on their role in liver diseases from acute injury to chronic inflammation, fibrosis and then to tumorigenesis. Notably, macrophage TAM receptor signalling seems to be a two-edged sword for liver diseases. On one hand, the activation of TAM receptor signalling inhibits inflammation and facilitates tissue repair during acute liver injury. On the other hand, continuous activation of the signalling contributes to the process of chronic inflammation into fibrosis and tumorigenesis by evoking hepatic stellate cells and inhibiting anti-tumour immunity. Therefore, targeting macrophage TAM receptors and clarifying its downstream pathways will be exciting prospects for the precaution and treatment of liver diseases, particularly at different stages or statuses.

Fish female-biased gene cyp19a1a leads to female antiviral response attenuation between sexes by autophagic degradation of MITA.

From insects to mammals, both innate and adaptive immune response are usually higher in females than in males, with the sex chromosome and hormonal differences considered the main reasons. Here, we report that zebrafish cyp19a1a (cytochrome P450, family 19, subfamily A, polypeptide 1a), an autosomal gene with female-biased expression, causes female fish to exhibit a lower antiviral response. First, we successfully constructed an infection model by intraperitoneal injection of spring viremia of carp virus (SVCV) into zebrafish (Danio rerio) and Carassius auratus herpesvirus (CaHV) in gibel carp (Carassius gibelio). Specifically, female fish were more vulnerable to viral infection than males, accompanied by a significantly weaker interferon (IFN) expression. After screening several candidates, cyp19a1a, which was highly expressed in female fish tissues, was selected for further analysis. The IFN expression and antiviral response were significantly higher in cyp19a1a-/- than in cyp19a1a+/+. Further investigation of the molecular mechanism revealed that Cyp19a1a targets mediator of IRF3 activation (MITA) for autophagic degradation. Interestingly, in the absence of MITA, Cyp19a1a alone could not elicit an autophagic response. Furthermore, the autophagy factor ATG14 (autophagy-related 14) was found interacted with Cyp19a1a to either promote or attenuate Cyp19a1a-mediated MITA degradation by either being overexpressed or knocked down, respectively. At the cellular level, both the normal and MITA-enhanced cellular antiviral responses were diminished by Cyp19a1a. These findings demonstrated a sex difference in the antiviral response based on a regulation mechanism controlled by a female-biased gene besides sex chromosome and hormonal differences, supplying the current understanding of sex differences in fish.

Zebrafish CERKL Enhances Host TBK1 Stability and Simultaneously Degrades Viral Protein via Ubiquitination Modulation.

In the viral infection process, host gene function is usually reported as either defending the host or assaulting the virus. In this study, we demonstrated that zebrafish ceramide kinase-like (CERKL) mediates protection against viral infection via two distinct mechanisms: stabilization of TANK-binding kinase 1 (TBK1) through impairing K48-linked ubiquitination and degradation of spring viremia of carp virus (SVCV) P protein by dampening K63-linked ubiquitination, resulting in an improvement of the host immune response and a decline in viral activity in epithelioma papulosum cyprini (EPC) cells. On SVCV infection, ifnφ1 expression was increased or blunted by CERKL overexpression or knockdown, respectively. Subsequently, we found that CERKL localized in the cytoplasm, where it interacted with TBK1 and enhanced its stability by impeding the K48-linked polyubiquitination; meanwhile, the antiviral capacity of TBK1 was significantly potentiated by CERKL. In contrast, CERKL also interacted with and degraded SVCV P protein to disrupt its function in viral proliferation. Further mechanism analysis revealed K63-linked deubiquitination is the primary means of CERKL-mediated SVCV P protein degradation. Taken together, our study reveals a novel mechanism of fish defense against viral infection: the single gene cerkl is both a shield for the host and a spear against the virus, which strengthens resistance.

Zebrafish Uba1 Degrades IRF3 through K48-Linked Ubiquitination to Inhibit IFN Production.

Fish IFN regulatory factor 3 (IRF3) is a crucial transcription factor in the IFN activation signaling pathway, which leads to IFN production and a positive cycle. Unrestricted IFN expression results in hyperimmune responses and therefore, IFN must be tightly regulated. In the current study, we found that zebrafish Ub-activating enzyme (Uba1) negatively regulated IRF3 via the K-48 ubiquitin proteasome degradation of IRF3. First, ifn expression stimulated by spring viraemia of carp virus infection was blunted by the overexpression of Uba1 and enhanced by Uba1 knockdown. Afterward, we found that Uba1 was localized in the cytoplasm, where it interacted with and degraded IRF3. Functional domains analysis revealed that the C-terminal ubiquitin-fold domain was necessary for IRF3 degradation by Uba1 and the N-terminal DNA-binding domain of IRF3 was indispensable for the degradation by Uba1.The degradation of IRF3 was subsequently impaired by treatment with MG132, a ubiquitin proteasome inhibitor. Further mechanism analysis revealed that Uba1 induced the K48-linked Ub-proteasomal degradation of IRF3. Finally, the antiviral capacity of IRF3 was significantly attenuated by Uba1. Taken together, our study reveals that zebrafish Uba1 interacts with and activates the ubiquitinated degradation of IRF3, providing evidence of the IFN immune balance mechanism in fish.

Electrospun Polymer-Free Nanofibers Incorporating Hydroxypropyl-ß-cyclodextrin/Difenoconazole via Supramolecular Assembly for Antifungal Activity.

In this study, flexible and self-standing hydroxypropyl-ß-cyclodextrin/difenoconazole inclusion complex (HPßCD/DZ-IC) nanofibers were prepared by polymer-free electrospinning, which exhibited potential to be a new fast-dissolving pesticide formulation. Scanning electron microscopy and optical microscopy were applied to evaluate the morphology of nanofibers, which showed that the resulting HPßCD/DZ-IC nanofibers were bead-free and uniform. In addition, the proton nuclear magnetic resonance (1H NMR) spectrum suggested a stoichiometric ratio of 1:0.9 (HPßCD/DZ). Other characterization methods, such as UV-vis absorption, fluorescence spectroscopy, Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), and thermogravimetric analysis (TGA), were applied in this study. On the one hand, UV-vis absorption, fluorescence spectroscopy, FT-IR, XRD, and TGA provided useful information for the successful formation of an inclusion complex; on the other hand, the results of TGA indicated the thermal stability of DZ was enhanced after the formation of inclusion complexes. Besides, the phase solubility test could explain the increased water solubility of the nanofibers of inclusion complexes formed by DZ and HPßCD. The results of molecular docking studies demonstrated the most favorable binding interactions when HPßCD combined with DZ. The dissolution test and the antifungal performance test exhibited the characteristics of fast dissolution and the excellent antifungal performance of HPßCD/DZ-IC nanofibers, respectively.

Grass Carp Reovirus VP35 Degrades MAVS Through the Autophagy Pathway to Inhibit Fish Interferon Production.

Fish interferon (IFN) is a crucial cytokine for a host to resist external pathogens, conferring cells with antiviral capacity. Meanwhile, grass carp reovirus (GCRV) is a strong pathogen that causes high mortality in grass carp. Therefore, it is necessary to study the strategy used by GCRV to evade the cellular IFN response. In this study, we found that GCRV 35-kDa protein (VP35) inhibited the host IFN production by degrading mitochondrial antiviral signaling (MAVS) protein through the autophagy pathway. First, the overexpression of VP35 inhibited the IFN activation induced by polyinosinic-polycytidylic acid (poly I:C) and MAVS, and the expression of downstream IFN-stimulated genes (ISGs) was also decreased by using VP35 under the stimulation. Second, VP35 interacted with MAVS; the experiments of truncated mutants of MAVS demonstrated that the caspase recruitment domain (CARD) and proline-rich (PRO) domains of MAVS were not necessary for this binding. Then, MAVS was degraded by using VP35 in a dose-dependent manner, and 3-MA (the autophagy pathway inhibitor) significantly blocked the degradation, meaning that MAVS was degraded by using VP35 in the autophagy pathway. The result of MAVS degradation suggested that the antiviral capacity of MAVS was remarkably depressed when interrupted by VP35. Finally, in the host cells, VP35 reduced ifn transcription and made the cells vulnerable to virus infection. In conclusion, our results reveal that GCRV VP35 impairs the host IFN response by degrading MAVS through the autophagy pathway, supplying evidence of a fish virus immune evasion strategy.

Molecular structure characterization of bituminous coal in Northern China via XRD, Raman and FTIR spectroscopy.

Bituminous coal is used widely for a variety of applications despite causing a range of problems within processes. The complexity and heterogeneity of the molecular structure of coal is one of the reasons for problems during use. Investigation into the molecular structure of the bituminous coal is reported from using X-ray diffraction (XRD), Raman spectroscopy, and Fourier Transform infrared (FTIR) spectroscopy experiments on four coal samples from coal mines in Northern China. The average lateral sizes (La), stacking heights (Lc) and interlayer spacing (d002) of the coal samples' crystallite structures derived from the XRD ranged from 25.78 to 27.93 Å, 17.27 to 25.88 Å and 3.40 to 3.52 Å, respectively; and the G-D1, ID1/IG and La of the samples ranged from 245.06 to 249.63 cm-1, 2.18 to 2.48 and 18.16 to 20.64 Å, respectively. The FTIR spectra reveals that coal samples incorporate oxygen-containing functional groups, aliphatic functional groups, aromatic functional groups and hydroxyl functional groups. Results show these four coal samples contained a low degree of ordered microcrystalline units with a low degree of aromatic conformation. The samples have the largest proportion of oxygenated functional groups, followed by aromatic structures, aliphatic structures and hydroxyl groups. Results from this study could inform the ongoing study of molecular structural characteristics of bituminous coal as well as help our understanding of properties such as wettability and pore structure.

Thiram/hydroxypropyl-ß-cyclodextrin inclusion complex electrospun nanofibers for a fast dissolving water-based drug delivery system.

The electrospinning of thiram/hydroxypropyl-ß-cyclodextrin inclusion complex nanofiber (thiram/HPßCD-IC-NF) was produced for establishing a quick dissolving water-based drug delivery system. As a dithiocarbamate broad-spectrum fungicide, thiram is insoluble in water. High-concentration HPßCD solutions (180 %, w/v) were applied in thiram/HPßCD systems to implement electrospinning with no extra polymer matrix added. The formation of thiram/HPßCD-IC-NF was identified by Fourier transform infrared spectroscopy, X-ray diffraction as well as nuclear magnetic resonance. Phase solubility study proved HPßCD played a huge role in the improvement in solubility of thiram, and thiram/HPßCD-IC-NF showed an excellent dissolution rate. Scanning electron microscopy was used to examine the configuration of surface of thiram/HPßCD-IC-NF, which exhibited that thiram/HPßCD-IC-NF was uniform and beadless. In addition, thiram/HPßCD-IC-NF exhibited better antifungal activity and thermal stability than pure thiram. In summary, thiram/HPßCD-IC-NF drug delivery system contributed to water solubility, thermal stability and antifungal activity of thiram. It could provide a new idea for the development of new formulations of rapidly dissolving green pesticides, and made efforts to promote the sustainable development of agriculture.

A novel role of Zebrafish TMEM33 in negative regulation of interferon production by two distinct mechanisms.

The transmembrane protein 33 (TMEM33) was originally identified as an endoplasmic reticulum (ER) protein that influences the tubular structure of the ER and modulates intracellular calcium homeostasis. However, the role of TMEM33 in antiviral immunity in vertebrates has not been elucidated. In this article, we demonstrate that zebrafish TMEM33 is a negative regulator of virus-triggered interferon (IFN) induction via two mechanisms: mitochondrial antiviral signaling protein (MAVS) ubiquitination and a decrease in the kinase activity of TANK binding kinase 1 (TBK1). Upon stimulation with viral components, tmem33 was remarkably upregulated in the zebrafish liver cell line. The IFNφ1 promoter (IFNφ1pro) activity and mRNA level induced by retinoic acid-inducible gene (RIG)-I-like receptors (RLRs) were significantly inhibited by TMEM33. Knockdown of TMEM33 increased host ifn transcription. Subsequently, we found that TMEM33 was colocalized in the ER and interacted with the RLR cascades, whereas MAVS was degraded by TMEM33 during the K48-linked ubiquitination. On the other hand, TMEM33 reduced the phosphorylation of mediator of IFN regulatory factor 3 (IRF3) activation (MITA)/IRF3 by acting as a decoy substrate of TBK1, which was also phosphorylated. A functional domain assay revealed that the N-terminal transmembrane domain 1 (TM1) and TM2 regions of TMEM33 were necessary for IFN suppression. Finally, TMEM33 significantly attenuated the host cellular antiviral capacity by blocking the IFN response. Taken together, our findings provide insight into the different mechanisms employed by TMEM33 in cellular IFN-mediated antiviral process.