Beta-Cell Tipe1 Orchestrates Insulin Secretion and Cell Proliferation by Promoting Gαs/cAMP Signaling via USP5.

Inadequate ß-cell mass and insulin secretion are essential for the development of type 2 diabetes (T2D). TNF-α-induced protein 8-like 1 (Tipe1) plays a crucial role in multiple diseases, however, a specific role in T2D pathogenesis remains largely unexplored. Herein, Tipe1 as a key regulator in T2D, contributing to the maintenance of ß cell homeostasis is identified. The results show that the ß-cell-specific knockout of Tipe1 (termed Ins2-Tipe1BKO) aggravated diabetic phenotypes in db/db mice or in mice with high-fat diet-induced diabetes. Notably, Tipe1 improves ß cell mass and function, a process that depends on Gαs, the α subunit of the G-stimulating protein. Mechanistically, Tipe1 inhibited the K48-linked ubiquitination degradation of Gαs by recruiting the deubiquitinase USP5. Consequently, Gαs or cAMP agonists almost completely restored the dysfunction of ß cells observed in Ins2-Tipe1BKO mice. The findings characterize Tipe1 as a regulator of ß cell function through the Gαs/cAMP pathway, suggesting that Tipe1 may emerge as a novel target for T2D intervention.

The Role of STAT3 Signaling Pathway Activation in Subconjunctival Scar Formation after Glaucoma Filtration Surgery.

Scar formation resulting from overly active wound healing is a critical factor in the success rate of glaucoma filtration surgery (GFS). IL-6 and TGF-ß have been implicated in the pathogenesis of fibrogenesis. In addition, the signal transducer and activator of transcription 3 (STAT3) can be activated by numerous cytokines and growth factors, including IL-6 and TGF-ß1. Thus, STAT3 activation may integrate common profibrotic pathways to promote fibrosis. In this study, an increase in p-STAT3 was observed in activated HTFs. Inhibiting STAT3 in cultured HTFs by pharmacological inactivation reversed the fibrotic responses, such as fibroblast migration, the differentiation of resting fibroblasts into myofibroblasts and the deposition of ECM, mediated by IL-6 and TGF-ß1. Moreover, the expression of suppressor of cytokine signaling 3 (SOCS3) was decreased in HTFs cultured with IL-6 and TGF-ß1, and SOCS3 overexpression rescued ECM deposition, α-SMA expression and migration in IL-6- and TGF-ß1-stimulated HTFs by inactivating STAT3. Finally, S3I-201 treatment inhibited profibrotic gene expression and subconjunctival fibrosis in a rat model of GFS. In conclusion, our data suggests that STAT3 plays a central role in fibrosis induced by different profibrotic pathways and that STAT3 is a potential target for antifibrotic therapies following GFS.

Large diurnal compensatory effects mitigate the response of Amazonian forests to atmospheric warming and drying.

Photosynthesis and evapotranspiration in Amazonian forests are major contributors to the global carbon and water cycles. However, their diurnal patterns and responses to atmospheric warming and drying at regional scale remain unclear, hindering the understanding of global carbon and water cycles. Here, we used proxies of photosynthesis and evapotranspiration from the International Space Station to reveal a strong depression of dry season afternoon photosynthesis (by 6.7 ± 2.4%) and evapotranspiration (by 6.1 ± 3.1%). Photosynthesis positively responds to vapor pressure deficit (VPD) in the morning, but negatively in the afternoon. Furthermore, we projected that the regionally depressed afternoon photosynthesis will be compensated by their increases in the morning in future dry seasons. These results shed new light on the complex interplay of climate with carbon and water fluxes in Amazonian forests and provide evidence on the emerging environmental constraints of primary productivity that may improve the robustness of future projections.

Management of angle-closure glaucoma with X-linked retinoschisis: a case report.

BACKGROUND: X-linked retinoschisis (XLRS), due to mutations in the RS1 gene, is a common genetically determined form of macular degeneration. This report describes an unusual case of angle-closure glaucoma (ACG) with XLRS and discusses the treatment. CASE PRESENTATION: A 39-year-old Chinese man with an X chromosome-recessive inherited c.489G > A variant in the RS1 gene was diagnosed as XLRS and ACG, presenting with cystic macular lesions, shallow anterior chamber depth (ACD), and angle-closure with uncontrolled intraocular pressure (IOP). Malignant glaucoma occurred following trabeculectomy combining phacoemulsification with intraocular lens (IOL) implantation and goniosynechialysis. Subsequent anterior vitrectomy and irido-zonulo-hyaloid-vitrectomy (IZHV) effectively lowered IOP and deepened ACD, but the cystic cavity became larger. CONCLUSIONS: There is a potential risk of malignant glaucoma in ACG patients with XLRS after filtering surgery. Although anterior vitrectomy can effectively resolve aqueous misdirection, the macular retinoschisis may get worse. Awareness of this risk may aid in surgical planning and postoperative management in these patients.

Protein Kinase C Modulation Determines the Mesoderm/Extraembryonic Fate Under BMP4 Induction From Human Pluripotent Stem Cells.

The interplay among mitogenic signaling pathways is crucial for proper embryogenesis. These pathways collaboratively act through intracellular master regulators to determine specific cell fates. Identifying the master regulators is critical to understanding embryogenesis and to developing new applications of pluripotent stem cells. In this report, we demonstrate protein kinase C (PKC) as an intrinsic master switch between embryonic and extraembryonic cell fates in the differentiation of human pluripotent stem cells (hPSCs). PKCs are essential to induce the extraembryonic lineage downstream of BMP4 and other mitogenic modulators. PKC-alpha (PKCα) suppresses BMP4-induced mesoderm differentiation, and PKC-delta (PKCδ) is required for trophoblast cell fate. PKC activation overrides mesoderm induction conditions and leads to extraembryonic fate. In contrast, PKC inhibition leads to ß-catenin (CTNNB1) activation, switching cell fate from trophoblast to mesoderm lineages. This study establishes PKC as a signaling boundary directing the segregation of extraembryonic and embryonic lineages. The manipulation of intrinsic PKC activity could greatly enhance cell differentiation under mitogenic regulation in stem cell applications.

Ubiquitous light-emitting diodes: Potential threats to retinal circadian rhythms and refractive development.

The use of light-emitting diodes (LEDs) has increased considerably in the 21st century with humans living in a modern photoperiod with brighter nights and dimmer days. Prolonged exposure to LEDs, especially at night, is considered a new source of pollution because it may affect the synthesis and secretion of retinal melatonin and dopamine, resulting in negative impacts on retinal circadian clocks and potentially disrupting retinal circadian rhythms. The control of ocular refraction is believed to be related to retinal circadian rhythms. Moreover, the global prevalence of myopia has increased at an alarming rate in recent decades. The widespread use of LEDs and the rapid increase in the prevalence of myopia overlap, which is unlikely to be a coincidence. The connection among LEDs, retinal circadian rhythms, and refractive development is both fascinating and confusing. In this review, we aim to develop a systematic framework that includes LEDs, retinal circadian rhythms and refractive development. This paper summarizes the possible mechanisms by which LEDs may disrupt retinal circadian rhythms. We propose that prolonged exposure to LEDs may induce myopia by disrupting retinal circadian rhythms. Finally, we suggest several possible countermeasures to prevent LED interference on retinal circadian rhythms, with the hope of reducing the onset and progression of myopia.

An Update on Novel Ocular Nanosystems with Possible Benefits in the Treatment of Corneal Neovascularization.

Corneal neovascularization (CNV) is an ocular pathological change that results from an imbalance between angiogenic factors and antiangiogenic factors as a result of various ocular insults, including infection, inflammation, hypoxia, trauma, corneal degeneration, and corneal transplantation. Current clinical strategies for the treatment of CNV include pharmacological treatment and surgical intervention. Despite some degree of success, the current treatment strategies are restricted by limited efficacy, adverse effects, and a short duration of action. Recently, gene-based antiangiogenic therapy has become an emerging strategy that has attracted considerable interest. However, potential complications with the use of viral vectors, such as potential genotoxicity resulting from long-term expression and nonspecific targeting, cannot be ignored. The use of ocular nanosystems (ONS) based on nanotechnology has emerged as a great advantage in ocular disease treatment during the last two decades. The potential functions of ONS range from nanocarriers, which deliver drugs and genes to target sites in the eye, to therapeutic agents themselves. Various preclinical studies conducted to date have demonstrated promising results of the use of ONS in the treatment of CNV. In this review, we provide an overview of CNV and its current therapeutic strategies and summarize the properties and applications of various ONS related to the treatment of CNV reported to date. Our goal is to provide a comprehensive review of these considerable advances in ONS in the field of CNV therapy over the past two decades to fill the gaps in previous related reports. Finally, we discuss existing challenges and future perspectives of the use of ONS in CNV therapy, with the goal of providing a theoretical contribution to facilitate future practical growth in the area.

Enhanced Delivery of TLR7/8 Agonists by Metal-Organic Frameworks for Hepatitis B Virus Cure.

Hepatitis B virus (HBV) infection remains a major challenge to global health due to unsatisfactory treatment efficacy, side effects of current therapies, and immune tolerance. Toll-like receptors 7/8 (TLR7/8) agonists have shown great potential in chronic hepatitis B (CHB) cure, but systemic administration often induces severe side effects due to rapid dispersion into the microvasculature. Herein, we encapsulate an imidazoquinoline-based TLR7/8 agonist (IMDQ) into zeolitic imidazolate framework 8 nanoparticles (IMDQ@ZIF-8 NPs) for HBV immunotherapy. Compared with free IMDQ, IMDQ@ZIF-8 NPs efficiently accumulate in the liver and are selectively taken up by antigen-presenting cells (APCs), leading to enhanced APC activation and efficient viral elimination in HBV-infected models. Strikingly, MDQ@ZIF-8 NP treatment results in the obvious production of anti-HBs antibody and seroconversion in HBV-infected mice. Overall, this study on the convergence of a facile assembly approach and efficient therapeutic effects represents a promising strategy for HBV treatment.

Label-Free Detection of DNA via Surface-Enhanced Raman Spectroscopy Using Au@Ag Nanoparticles.

DNA is a building block of life; surface-enhanced Raman spectroscopy (SERS) has been broadly applied in the detection of biomolecules but there are challenges in obtaining high-quality DNA SERS signals under non-destructive conditions. Here, we developed a novel label-free approach for DNA detection based on SERS, in which the Au@AgNPs core-shell structure was selected as the enhancement substrate, which not only solved the problem of the weak enhancement effect of gold nanoparticles but also overcame the disadvantage of the inhomogeneous shapes of silver nanoparticles, thereby improving the sensitivity and reproducibility of the SERS signals of DNA molecules. The method obtained SERS signals for four DNA bases (A, C, G, and T) without destroying the structure, then further detected and qualified different specific structures of DNA molecules. These results promote the application of SERS technology in the field of biomolecular detection.

Histidine acid phosphatase domain-containing protein from Haemonchus contortus is a stimulatory antigen for the Th1 immune response of goat PBMCs.

BACKGROUND: Histidine acid phosphatase (HAP), a member of the histidine phosphatase superfamily, is widely found in parasites and is also a potential vaccine antigen or drug target. However, the biological function of HAP in Haemonchus contortus is still unclear. METHODS: We cloned the HAP gene from H. contortus (Hc-HAP) and expressed the purified recombinant Hc-HAP (rHc-HAP) protein. The transcription of the Hc-HAP gene in the eggs, infective third-stage larvae (L3s), exsheathed third-stage larvae (xL3s) and adults (females/males) was analyzed by quantitative real-time-PCR (qPCR). An immunofluorescence assay was also used to detect the localization of Hc-HAP expression in adult worms. The effect of rHc-HAP on the function of peripheral blood mononuclear cells (PBMCs) was observed by co-culture of rHc-HAP protein with goat PBMCs. RESULTS: The qPCR results revealed that the Hc-HAP gene was transcribed at a higher level in the L3 and xL3 stages that there were gender differences in transcription at the adult stage, with females exhibiting higher transcription than males. Moreover, Hc-HAP was mainly expressed in adult intestinal microvilli. Additionally, western blot results revealed that rHc-HAP could be detected in goat sera artificially infected with H. contortus. In the experiments, rHc-HAP bound to goat PBMCs and released nitric oxide. The rHc-HAP also induced the expression of interferon gamma (IFN-γ) and the phosphorylated STAT 1 transcription factor, while inhibiting interleukin-4 expression. CONCLUSIONS: The results shows that rHc-HAP stimulated the IFN-γ/STAT1 signaling pathway and enabled polarization of PBMCs toward T-helper 1 immune responses.

cccDNA Surrogate MC-HBV-Based Screen Identifies Cohesin Complex as a Novel HBV Restriction Factor.

BACKGROUND & AIMS: Covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV), existing as a stable minichromosome in the hepatocyte, is responsible for persistent HBV infection. Maintenance and sustained replication of cccDNA require its interaction with both viral and host proteins. However, the cccDNA-interacting host factors that limit HBV replication remain elusive. METHODS: Minicircle HBV (MC-HBV), a recombinant cccDNA, was constructed based on chimeric intron and minicircle DNA technology. By mass spectrometry based on pull-down with biotinylated MC-HBV, the cccDNA-hepatocyte interaction profile was mapped. HBV replication was assessed in different cell models that support cccDNA formation. RESULTS: MC-HBV supports persistent HBV replication and mimics the cccDNA minichromosome. The MC-HBV-based screen identified cohesin complex as a cccDNA binding host factor, leading to reduced HBV replication. Mechanistically, with the help of CCCTC-binding factor (CTCF), which has specific binding sites on cccDNA, cohesin loads on cccDNA and reshapes cccDNA confirmation to prevent RNA polymerase II enrichment. Interestingly, HBV X protein transcriptionally reduces structural maintenance of chromosomes complex expression to partially relieve the inhibitory role of the cohesin complex on HBV replication. CONCLUSIONS: Our data not only provide a feasible approach to explore cccDNA-binding factors, but also identify cohesin/CTCF complex as a critical host restriction factor for cccDNA-driven HBV replication. These findings provide a novel insight into cccDNA-host interaction and targeted therapeutic intervention for HBV infection.

Insulin Directs Dichotomous Translational Regulation to Control Human Pluripotent Stem Cell Survival, Proliferation and Pluripotency.

Insulin is essential for diverse biological processes in human pluripotent stem cells (hPSCs). However, the underlying mechanism of insulin's multitasking ability remains largely unknown. Here, we show that insulin controls hPSC survival and proliferation by modulating RNA translation via distinct pathways. It activates AKT signaling to inhibit RNA translation of pro-apoptotic proteins such as NOXA/PMAIP1, thereby promoting hPSC survival. At the same time, insulin acts via the mTOR pathway to enhance another set of RNA translation for cell proliferation. Consistently, mTOR inhibition by rapamycin results in eIF4E phosphorylation and translational repression. It leads to a dormant state with sustained pluripotency but reduced cell growth. Together, our study uncovered multifaceted regulation by insulin in hPSC survival and proliferation, and highlighted RNA translation as a key step to mediate mitogenic regulation in hPSCs.

LINC01431 Promotes Histone H4R3 Methylation to Impede HBV Covalently Closed Circular DNA Transcription by Stabilizing PRMT1.

Covalently closed circular DNA (cccDNA) is the transcriptional template of hepatitis B virus (HBV), which interacts with both host and viral proteins to form minichromosome in the nucleus and is resistant to antiviral agents. Identification of host factors involved in cccDNA transcriptional regulation is expected to prove a new venue for HBV therapy. Recent evidence suggests the involvement of long noncoding RNAs (lncRNAs) in mediating the interaction of host factors with various viruses, however, lncRNAs that HBV targets and represses cccDNA transcription have not been fully elucidated. Here, the authors identified LINC01431 as a novel host restriction factor for HBV transcription. Mechanically, LINC01431 competitively bound with type I protein arginine methyltransferase (PRMT1) to block the HBx-mediated PRMT1 ubiquitination and degradation. Consequently, LINC01431 increased the occupancy of PRMT1 on cccDNA, leading to enhanced H4R3me2a modification and reduced acetylation of cccDNA-bound histones, thereby repressing cccDNA transcription. In turn, to facilitate viral replication, HBV transcriptionally repressed LINC01431 expression by HBx-mediated repression of transcription factor Zinc fingers and homeoboxes 2 (ZHX2). Collectively, the study demonstrates LINC01431 as a novel epigenetic regulator of cccDNA minichromosome and highlights a feedback loop of HBx-LINC01431-PRMT1 in HBV replication, which provides potential therapeutic targets for HBV treatment.

Thyroid hormone enhances stem cell maintenance and promotes lineage-specific differentiation in human embryonic stem cells.

BACKGROUND: Thyroid hormone triiodothyronine (T3) is essential for embryogenesis and is commonly used during in vitro fertilization to ensure successful implantation. However, the regulatory mechanisms of T3 during early embryogenesis are largely unknown. METHOD: To study the impact of T3 on hPSCs, cell survival and growth were evaluated by measurement of cell growth curve, cloning efficiency, survival after passaging, cell apoptosis, and cell cycle status. Pluripotency was evaluated by RT-qPCR, immunostaining and FACS analysis of pluripotency markers. Metabolic status was analyzed using LC-MS/MS and Seahorse XF Cell Mito Stress Test. Global gene expression was analyzed using RNA-seq. To study the impact of T3 on lineage-specific differentiation, cells were subjected to T3 treatment during differentiation, and the outcome was evaluated using RT-qPCR, immunostaining and FACS analysis of lineage-specific markers. RESULTS: In this report, we use human pluripotent stem cells (hPSCs) to show that T3 is beneficial for stem cell maintenance and promotes trophoblast differentiation. T3 enhances culture consistency by improving cell survival and passaging efficiency. It also modulates cellular metabolism and promotes energy production through oxidative phosphorylation. T3 helps maintain pluripotency by promoting ERK and SMAD2 signaling and reduces FGF2 dependence in chemically defined culture. Under BMP4 induction, T3 significantly enhances trophoblast differentiation. CONCLUSION: In summary, our study reveals the impact of T3 on stem cell culture through signal transduction and metabolism and highlights its potential role in improving stem cell applications.

Autophagy inhibition by TSSC4 (tumor suppressing subtransferable candidate 4) contributes to sustainable cancer cell growth.

Cancer cell growth is dependent upon the sustainability of proliferative signaling and resisting cell death. Macroautophagy/autophagy promotes cancer cell growth by providing nutrients to cells and preventing cell death. This is in contrast to autophagy promoting cell death under some conditions. The mechanism regulating autophagy-mediated cancer cell growth remains unclear. Herein, we demonstrate that TSSC4 (tumor suppressing subtransferable candidate 4) is a novel tumor suppressor that suppresses cancer cell growth and tumor growth and prevents cell death induction during excessive growth by inhibiting autophagy. The oncogenic proteins ERBB2 (erb-b2 receptor tyrosine kinase 2) and the activation EGFR mutant (EGFRvIII, epidermal growth factor receptor variant III) promote cell growth and TSSC4 expression in breast cancer and glioblastoma multiforme (GBM) cells, respectively. In EGFRvIII-expressing GBM cells, TSSC4 knockout shifted the function of autophagy from a pro-cell survival role to a pro-cell death role during prolonged cell growth. Furthermore, the interaction of TSSC4 with MAP1LC3/LC3 (microtubule associated protein 1 light chain 3) via its conserved LC3-interacting region (LIR) contributes to its inhibition of autophagy. Finally, TSSC4 suppresses tumorsphere formation and tumor growth by inhibiting autophagy and maintaining cell survival in tumorspheres. Taken together, sustainable cancer cell growth can be achieved by autophagy inhibition via TSSC4 expression.Abbreviations: 3-MA: 3-methyladenine; ACTB: actin beta; CQ: chloroquine; EGFRvIII: epidermal growth factor receptor variant III; ERBB2: erb-b2 receptor tyrosine kinase 2; GBM: glioblastoma multiforme; LIR: LC3-interacting region; MAP1LC3/LC3: microtubule Associated protein 1 light chain 3; TSSC4: tumor suppressing subtransferable candidate 4.

Tracking the seasonal and inter-annual variations of global gross primary production during last four decades using satellite near-infrared reflectance data.

Terrestrial vegetation absorbs approximately 30% of the anthropogenic carbon dioxide (CO2) emitted into the atmosphere through photosynthesis (represented by gross primary productivity, GPP) and thus effectively mitigates global warming. However, large uncertainties still remain in the global GPP estimations and their long-term trends. Here we used the satellite-based near-infrared reflectance (NIRv) as the proxy of GPP and generated a global long-term (1982-2018) GPP datasets (hereafter GPPNIRv). Analysis at the site-level showed that NIRv could accurately capture both the monthly and annual variations in GPP (R2 = 0.71 and 0.74 respectively) at 104 flux sites. Upscaling the relationships between NIRv and GPP to the global scale, the global annual GPP was estimated to be 128.3 ± 4.0 Pg C yr-1 during the last four decades, which fell between the estimations from the machine-learning upscaling approach, light-use-efficiency (LUE) models and processed-based models. The seasonal variation of GPPNIRv was also consistent with those from flux sites and models. More importantly, the inter-annual trends in GPPNIRv during the last four decades were consistent with those from processed-based models across latitudes, which outperformed the other GPP products. This evidence suggested that the long-term GPP datasets derived from NIRv have better abilities to capture the seasonal and inter-annual variations of terrestrial GPP at the global scale. The long-term GPPNIRv product could be beneficial for the estimation of terrestrial carbon fluxes and for the projection of future climates.

Seasonal variations in the relationship between sun-induced chlorophyll fluorescence and photosynthetic capacity from the leaf to canopy level in a rice crop.

Photosynthetic capacity (leaf maximum carboxylation rate, Vcmax) is a critical parameter for accurately assessing carbon assimilation by plant canopies. Recent studies of sun-induced chlorophyll fluorescence (SIF) have shown potential for estimating Vcmax at the ecosystem level. However, the relationship between SIF and Vcmax at the leaf and canopy levels is still poorly understood. In this study, we investigated the dynamic relationship between SIF and Vcmax and its controlling factors using SIF and CO2 response measurements in a rice paddy. We found that SIF and its yield (SIFy) were strongly correlated with Vcmax during the growing season, although the relationship varied with plant growth stages. After flowering, SIFy showed a stronger relationship with Vcmax than SIF flux at both the leaf and canopy levels. Further analysis suggested that the divergence of the link between SIF and Vcmax from leaf to canopy are the result of changes in canopy structure and leaf physiology, highlighting that these need to be considered when interpreting the SIF signal across spatial scales. Our results provide evidence that remotely sensed SIF observations can be used to track seasonal variations in Vcmax at the leaf and canopy levels.

Grassland dynamics in responses to climate variation and human activities in China from 2000 to 2013.

Improving our understanding of the impacts of climate variation and human activities on grassland dynamics is heightened by expectations that climate variation and human activities may induce grassland degradation. An accurate evaluation of the respective impacts of climate variation and human activities on grassland dynamics is crucial to understand the grassland degradation mechanism and to control the degraded grassland. In this study, net primary productivity (NPP) was selected as an indicator to reflect grassland dynamics. Meanwhile, the potential NPP (PNPP) and human-induced NPP (HNPP) calculated as the difference of PNPP and actual ANPP (ANPP) were used to assess the relative effects of climate variation and human activities on grassland NPP changes in China during 2000-2013. Results of grassland ANPP showed an overall increase than decrease in productivity (81.21% vs 18.79%) from 2000 to 2013. For the increase of ANPP, the relative contribution of climate variation and human activities to grassland NPP changes were 41.45% and 45.22%, respectively. Climate variation was the dominant factor that induced the increase in ANPP mainly in areas of Sichuan, Gansu, Ningxia and Inner Mongolia. An increase in Human-dominated ANPP mainly occurred in Tibet, Qinghai and Xinjiang. The decrease in ANPP is principally controlled by the effect of human activities than that of climate variation, especially in Inner Mongolia. Meanwhile, climate-dominated ANPP increase and human-dominated ANPP decrease mainly occurred in plain grassland, desert grassland and meadow across the six types of grasslands in China. Furthermore, in alpine sub-alpine meadow and alpine sub-alpine, while climate-dominated ANPP of grassland was found to be decreased, an increase in human-dominated ANPP was detected. The increase in precipitation and the implementation of ecological restoration programs were found to be effective in inducing the noticeable increased grassland ANPP since 2003. The findings of the current research recommend that the Chinese government should continue to implement the prohibiting graze policy across the country and extensively strengthen the implementation of the policy in Inner Mongolia and North Xinjiang, particularly in plain grassland, desert grassland and meadow.