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Purpose: Gadolinium-neutron capture therapy (Gd-NCT) employs isotopically enriched Gadolinium (Gd) and thermal neutrons to selectively target cancer cells. This study investigated the targeting efficacy of 157Gd-DOTA-PSMA (Prostate-Specific Membrane Antigen) in prostate cancer and explored its potential applications in Gd-NCT. Methods and results: We developed 157Gd-DOTA-PSMA, a novel theranostic bio-gadolinium agent specifically designed for magnetic resonance imaging (MRI)-guided Gd-NCT. 68 Ga-DOTA-PSMA positron emission tomography-computed tomography (PET/CT) imaging showed peak radiotracer uptake at 2 h post-injection, with a tumor-to-non-tumor (T/NT) ratio of 6.95 ± 0.60. MRI analysis confirmed a stable T1 signal enhancement 2 h post-injection. Time-of-flight inductively coupled plasma mass spectrometry (TOF-ICP-MS) revealed significantly elevated Gd concentrations in 22Rv1 tumor compared to PC-3 tumor and other healthy organs. ICP-MS analysis showed Gd concentrations of 165.69 μg [Gd]/g in 22Rv1 tumors and 35.25 μg [Gd]/g in blood, yielding a tumor-to-blood (T/B) ratio of 4.65 ± 0.54 and a T/NT ratio of 3.65 ± 0.49. Neutron irradiation with 157Gd-DOTA-PSMA reduced cell viability, inhibited colony formation, and induced DNA damage and apoptosis in 22Rv1 cells. In 22Rv1 mice, γ-H2AX levels peaked at 6 h post-irradiation, accompanied by an increase in pro-apoptotic proteins and a decrease in anti-apoptotic proteins over 24 h. In the NCT group following the injection of 157Gd-DOTA-PSMA, there was effective suppression of tumor growth without a loss of body weight, resulting in a 1.7-fold increase in median survival compared to control group. Conclusions: 157Gd-DOTA-PSMA, as a theranostic bio-gadolinium agent designed for targeted Gd-NCT in prostate cancer, represents a novel therapeutic approach and broadens the scope of potential applications of neutron capture therapy. [ABSTRACT FROM AUTHOR]

【Objective】 Evaporation loss via capillary rise is an important component of groundwater cycle but difficult to measure in situ. This paper presents an empirical formula to estimate it using surface evaporation and ambient air temperature, based on long-term experimental data obtained from a hydrological experimental station. 【Method】 Groundwater evaporation from phreatic surface of shallow groundwater was measured from lysimeters with cross section of 0.3 m2 at Wudaogou Hydrological Experiment Station, and the meteorological data were measured from a weather station installed at the station. There were two soils, a fluvo-aquic soil and a lime concretion black soil. For each soil, we established a regression model using temperature and surface evaporation, either separately or in combination, to estimate groundwater evaporation. 【Result】 For the lime concretion black soil, the average R2 of the models using single or two meteorological factors was 0.88, and its associated average absolute error (MAE) and root mean square error (RMSE) were 0.15 mm/d and 0.23 mm/d, respectively. For the yellow fluvo-aquic soil, the associated results were R2=0.92, MAE=0.194 mm, and RMSE=0.28 mm/d. When using a single meteorological factor, the average R2 for the two soils using all models was R2=0.92, and its associated MAE and RMSE were 0.18 mm/d and 0.26 mm/d, respectively. In contrast, in using the two meteorological factors, the average R2 for the two soils using all models was 0.94, and its associated MAE and RMSE were 0.14 mm/d and 0.205 mm/d, respectively. 【Conclusion】 For a given area, the evaporation of its shallow groundwater via capillary rise depends on surface evaporation and ambient air temperature. Since air temperature and surface evaporation are closely correlated, we can use either one of the two factor or their combination to estimate the groundwater evaporation.

Marek's Disease (MD), caused by Marek's disease virus (MDV), is a highly contagious lymphoproliferative disease in poultry. Despite the fact that MD has been effectively controlled by vaccines, the virulence of field isolates of MDV has continued to evolve, becoming more virulent under the immune pressure of vaccines. Our previous research has confirmed that the recombinant rMDV strain with REV-LTR insertion can be used as a live attenuated vaccine candidate. The aim of this research was to evaluate the onset and duration of immunity of the rMDV strain through two experiments. In both experiments, 1-day-old SPF chickens were vaccinated subcutaneously with the rMDV strain at a dose of 3,000 Plaque Formation Unit (PFU) per chick in 0.2 mL of the MD diluent. Then, in Experimental design 1, the chicks in the groups Vac-3d/CC-3d, Vac-5d/CC-5d, and Vac-7d/CC-7d were challenged separately with 500 PFU vvMDV strain MD5 at 3 days, 5 days, and 7 days after vaccination; in Experimental design 2, the chicks in group Vac-60d/CC-60d, Vac-120d/CC-120d, and Vac-180d/CC-180d were challenged at 60 days, 120 days, and 180 days after vaccination. The clinical symptoms and weight gain of chickens in each group were observed and recorded. The results showed that the rMDV strain with REV-LTR insertion provides protection starting from 3 days of age and achieves good immune effects at 5 days of age after 1-day-old immunization, and the immunization duration can reach for at least 180 days. Given age-related resistance, it can be confirmed that our vaccine can actually provide lifelong immunity. This study provides valuable insights into the onset and duration of immunity of the rMDV strain, which will provide a basis for the development and improvement of MD vaccines. [ABSTRACT FROM AUTHOR]

Parthanatos is distinct from caspase-dependent apoptosis in that it does not necessitate the activation of caspase cascades; Instead, it relies on the translocation of Apoptosis-inducing Factor (AIF) from the mitochondria to the nucleus, resulting in nuclear DNA fragmentation. Newcastle Disease Virus (NDV) is an oncolytic virus that selectively targets and kills tumor cells by inducing cell apoptosis. It has been reported that NDV triggers classic apoptosis through the mitochondrial pathway. In this study, we observed that NDV infection induced endoplasmic reticulum stress (ERS), which caused a rapid release of endogenous calcium ions (Ca2+). This cascade of events resulted in mitochondrial depolarization, loss of mitochondrial membrane potential, and structural remodeling of the mitochondria. The overload of Ca2+ also initiated an increase in mitochondrial membrane permeability, facilitating the transfer of AIF to the nucleus to induce apoptosis. Damaged mitochondria produced excessive reactive oxygen species (ROS), which further exacerbated mitochondrial damage and increased mitochondrial membrane permeability, thus promoting additional intracellular Ca2+ accumulation and ultimately triggering an ROS burst. Collectively, these findings indicate that NDV infection promotes excessive calcium accumulation and ROS generation, leading to mitochondrial damage that releases more calcium and ROS, creating a feedback loop that exacerbates AIF-dependent parthanatos. This study not only provides a novel perspective on the oncolytic mechanism of NDV but also highlights new targets for antiviral research. Author summary: Apoptosis serves as a cellular self-defense mechanism against viral infections, while also being a consequence of viral proliferation within infected cells. Mitochondria play a critical role in amplifying the stress associated with viral infections, as they sustain damage in response to intracellular apoptosis-inducing signals. Our observations indicate that NDV infection leads to Ca2+ accumulation and excessive ROS production, which in turn results in mitochondrial depolarization, loss of membrane potential, increased permeability, and the subsequent release and transport of AIF to the nucleus, ultimately triggering tumor cell death. These findings provide compelling evidence for an NDV-induced parthanatos mechanism and underscore the significant roles of calcium and ROS in mitochondrial apoptotic pathways. [ABSTRACT FROM AUTHOR]

The hallmark of coronavirus infection lies in its ability to evade host immune defenses, a process intricately linked to the nuclear entry of transcription factors crucial for initiating the expression of antiviral genes. Central to this evasion strategy is the manipulation of the nucleocytoplasmic trafficking system, which serves as an effective target for the virus to modulate the expression of immune response-related genes. In this investigation, we discovered that infection with the infectious bronchitis virus (IBV) dynamically impedes the nuclear translocation of several transcription factors such as IRF3, STAT1, STAT2, NF-κB p65, and the p38 MAPK, leading to compromised transcriptional induction of key antiviral genes such as IFNβ, IFITM3, and IL-8. Further examination revealed that during the infection process, components of the nuclear pore complex (NPC), particularly FG-Nups (such as NUP62, NUP153, NUP42, and TPR), undergo cytosolic dispersion from the nuclear envelope; NUP62 undergoes phosphorylation, and NUP42 exhibits a mobility shift in size. These observations suggest a disruption in nucleocytoplasmic trafficking. Screening efforts identified the IBV nucleocapsid (N) protein as the agent responsible for the cytoplasmic distribution of FG-Nups, subsequently hindering the nuclear entry of transcription factors and suppressing the expression of antiviral genes. Interactome analysis further revealed that the IBV N protein interacts with the scaffold protein RACK1, facilitating the recruitment of activated protein kinase C alpha (p-PKCα) to RACK1 and relocating the p-PKCα-RACK1 complex to the cytoplasm. These observations are conserved across diverse coronaviruses N proteins. Concurrently, the presence of both RACK1 and PKCα/β proved essential for the phosphorylation and cytoplasmic dispersion of NUP62, the suppression of antiviral cytokine expression, and efficient virus replication. These findings unveil a novel, highly effective, and evolutionarily conserved mechanism. Author summary: Coronaviruses employ diverse strategies to suppress the host innate immune defense. In this study, we uncovered a novel and highly effective strategy utilized by diverse coronaviruses to inhibit the innate immune response. Specifically, we found that the coronavirus N protein facilitates the binding of p-PKCα to RACK1, leading to the phosphorylation of NUP62 and the cytoplasmic redistribution of multiple FG-Nups. This phenomenon is accompanied by the disruption of nuclear translocation of several innate immune response-related transcription factors and suppression of antiviral/pro-inflammatory gene expression. Our research represents the first elucidation of how the N protein targets and impairs NPC function through the promotion of p-PKCα- RACK1 interaction and NUP62 phosphorylation/disassembly. This discovery unveils a novel mechanism employed by diverse coronaviruses to counteract the host immune response. [ABSTRACT FROM AUTHOR]

Although the involvement of the ubiquitin-proteasome system (UPS) in several coronavirus-productive infections has been reported, whether the UPS is required for infectious bronchitis virus (IBV) and porcine epidemic diarrhea virus (PEDV) infections is unclear. In this study, the role of UPS in the IBV and PEDV life cycles was investigated. When the UPS was suppressed by pharmacological inhibition at the early infection stage, IBV and PEDV infectivity were severely impaired. Further study showed that inhibition of UPS did not change the internalization of virus particles; however, by using R18 and DiOC-labeled virus particles, we found that inhibition of UPS prevented the IBV and PEDV membrane fusion with late endosomes or lysosomes. In addition, proteasome inhibitors blocked the degradation of the incoming viral protein N, suggesting the uncoating process and genomic RNA release were suppressed. Subsequently, the initial translation of genomic RNA was blocked. Thus, UPS may target the virus-cellular membrane fusion to facilitate the release of incoming viruses from late endosomes or lysosomes, subsequently blocking the following virus uncoating, initial translation, and replication events. Similar to the observation of proteasome inhibitors, ubiquitin-activating enzyme E1 inhibitor PYR-41 also impaired the entry of IBV, enhanced the accumulation of ubiquitinated proteins, and depleted mono-ubiquitin. In all, this study reveals an important role of UPS in coronavirus entry by preventing membrane fusion and identifies UPS as a potential target for developing antiviral therapies for coronavirus. [ABSTRACT FROM AUTHOR]

Modeling methodologies capable of coping with changing catchment conditions are becoming more important and are critical to ensure credible model predictions because the hydrologic process of the catchment is inevitably affected by climate change and human activities. A recently developed hydrological modeling framework that treats model parameters to be time-variant by building functions based on identified catchment properties is applied to the Upper Ganjiang River Basin in China experienced soil and water conservation constructions. The efficacy of the time-variant parameter method is investigated, and we further explore the impacts of model structure on hydrological modeling through this method. The approach is tested using two conceptual monthly water balance models, i.e., the ab model derived from the abcd model, and a two-parameter monthly water balance model (TWBM), which have the same model inputs and outputs, number of parameters and state variables. Results of the case study show that through the comparative analysis with the constant parameter models, both time-variant parameter models provide improved runoff simulations, especially for low and high flows. Moreover, the time-variant parameter method brings relatively larger runoff improvements for the TWBM model. For the actual evapotranspiration simulations, the ab model can yield improvements by treating parameter b (i.e., upper soil zone water holding capacity) to be time-variant, whereas the time-variant parameter method shows no efficacy in that for the TWBM model. The results demonstrate that the choice of hydrological model plays an important role in modeling performance of the time-variant parameter framework under a changing environment. Modification of the evapotranspiration calculation module for the TWBM model could be considered as further study to investigate the potential model improvement that compared with only time-variant parameters. This study can provide beneficial reference to comprehensively understand the impacts of changing environment on catchment hydrological modeling and thus improve the regional strategy for future water resource management. [ABSTRACT FROM AUTHOR]

Lacking a self-contained metabolism network, viruses have evolved multiple mechanisms for rewiring the metabolic system of their host to hijack the host's metabolic resources for replication. Newcastle disease virus (NDV) is a paramyxovirus, as an oncolytic virus currently being developed for cancer treatment. However, how NDV alters cellular metabolism is still far from fully understood. In this study, we show that NDV infection reprograms cell metabolism by increasing glucose utilization in the glycolytic pathway. Mechanistically, NDV induces mitochondrial damage, elevated mitochondrial reactive oxygen species (mROS) and ETC dysfunction. Infection of cells depletes nucleotide triphosphate levels, resulting in elevated AMP:ATP ratios, AMP-activated protein kinase (AMPK) phosphorylation, and MTOR crosstalk mediated autophagy. In a time-dependent manner, NDV shifts the balance of mitochondrial dynamics from fusion to fission. Subsequently, PINK1-PRKN-dependent mitophagy was activated, forming a ubiquitin chain with MFN2 (mitofusin 2), and molecular receptor SQSTM1/p62 recognized damaged mitochondria. We also found that NDV infection induces NAD+-dependent deacetylase SIRT3 loss via mitophagy to engender HIF1A stabilization, leading to the switch from oxidative phosphorylation (OXPHOS) to aerobic glycolysis. Overall, these studies support a model that NDV modulates host cell metabolism through PINK1-PRKN-dependent mitophagy for degrading SIRT3. Abbreviations: AMPK: AMP-activated protein kinase; CCCP: carbonyl cyanide 3-chlorophenylhydrazone; ECAR: extracellular acidification rate; hpi: hours post infection LC-MS: liquid chromatography-mass spectrometry; mito-QC: mCherry-GFP-FIS1[mt101–152]; MFN2: mitofusin 2; MMP: mitochondrial membrane potential; mROS: mitochondrial reactive oxygen species; MOI: multiplicity of infection; 2-NBDG: 2-(N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl) amino)-2-deoxyglucose; NDV: newcastle disease virus; OCR: oxygen consumption rate; siRNA: small interfering RNA; SIRT3: sirtuin 3; TCA: tricarboxylic acid; TCID50: tissue culture infective doses. [ABSTRACT FROM AUTHOR]

Infectious bronchitis virus (IBV) has been prevalent in chicken farms for many years, and its control relies on extensive vaccine administration. The continuous emergence of new variants and the low cross-protection efficiency prompt the development of new vaccines. In this study, we develop a reverse genetics technique based on the classical vaccine strain H120 genome, via in vitro ligation method. Using the H120 genome as the backbone, we constructed the recombinant virus rH120-QX(S) by replacing the H120 S gene with the QX S gene, a prevalent strain in China. Biological characteristics of the rH120-QX(S) virus, such as 50% egg lethal dose (ELD50), 50% egg infectious dose (EID50), dwarf embryo, growth curve, and genetic stability, are measured, which are comparable to the parental virus H120. There are no clinical symptoms and tissue lesions in the trachea and kidney in the rH120-QX(S)-infected specific-pathogen-free (SPF) chickens, demonstrating that this recombinant virus does not confer pathogenicity. Furthermore, protection studies show that there is 100% homologous protection of rH120-QX(S) to the virulent QX strain, as shown by the absence of clinical signs and no lethality. Taken together, our results demonstrate that swapping the S gene onto the H120 genetic backbone is a precise and effective way to produce genetically defined IBV vaccine candidates. [ABSTRACT FROM AUTHOR]

Virus infection can lead to the production of interferon, which activates the JAK/STAT pathway and induces the expression of multiple downstream interferon-stimulated genes (ISGs) to achieve their antiviral function. Cytidine/uridine monophosphate kinase 2 (CMPK2) gene has been identified as an ISG in human and fish, and is also known as a rate-limiting enzyme in mitochondria to maintain intracellular UTP/CTP levels, which is necessary for de novo mitochondrial DNA synthesis. By mining previous microarray data, it was found that both Avian Influenza Virus (AIV) and Newcastle Disease Virus (NDV) infection can lead to the significant upregulation of chicken CMPK2 gene. However, little is known about the function of CMPK2 gene in chickens. In the present study, the open reading frame (ORF) of chicken CMPK2 (chCMPK2) was cloned from DF-1, a chicken embryo fibroblasts cell line, and subjected to further analysis. Sequence analysis showed that chCMPK2 shared high similarity in amino acid with CMPK2 sequences from all the other species, especially reptiles. A thymidylate kinase (TMK) domain was identified in the C-terminus of chCMPK2, which is highly conserved among all species. In vitro , AIV infection induced significant increases in chCMPK2 expression in DF-1, HD11, and the chicken embryonic fibroblasts (CEF), while obvious increase only detected in DF-1 cells and CEF cells after NDV infection. In vivo , the expression levels of chCMPK2 were up-regulated in several tissues from AIV infected chickens, especially the brain, spleen, bursa, kidney, intestine, heart and thymus, and notable increase of chCMPK2 was detected in the bursa, kidney, duodenum, lung, heart, and thymus during NDV infection. Here, using MDA5 and IFN-β knockdown cells, we demonstrated that as a novel ISG, chCMPK2 could be regulated by the MDA5/IFN-β pathway. The high expression level of exogenous chCMPK2 displayed inhibitory effects on AIV and NDV as well as reduced viral RNA in infected cells. We further demonstrated that Asp135, a key site on the TMK catalytic domain, was identified as critical for the antiviral activities of chCMPK2. Taken together, these data demonstrated that chCMPK2 is involved in the chicken immune system and may play important roles in host anti-viral responses. [ABSTRACT FROM AUTHOR]

Host cholesterol metabolism remodeling is significantly associated with the spread of human pathogenic coronaviruses, suggesting virus-host relationships could be affected by cholesterol-modifying drugs. Cholesterol has an important role in coronavirus entry, membrane fusion, and pathological syncytia formation, therefore cholesterol metabolic mechanisms may be promising drug targets for coronavirus infections. Moreover, cholesterol and its metabolizing enzymes or corresponding natural products exert antiviral effects which are closely associated with individual viral steps during coronavirus replication. Furthermore, the coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 infections are associated with clinically significant low cholesterol levels, suggesting cholesterol could function as a potential marker for monitoring viral infection status. Therefore, weaponizing cholesterol dysregulation against viral infection could be an effective antiviral strategy. In this review, we comprehensively review the literature to clarify how coronaviruses exploit host cholesterol metabolism to accommodate viral replication requirements and interfere with host immune responses. We also focus on targeting cholesterol homeostasis to interfere with critical steps during coronavirus infection. [ABSTRACT FROM AUTHOR]

Abstract A TaqMan-based real-time polymerase chain reaction (PCR) assay was devised for the detection of porcine parvovirus (PPV). Two primers and a TaqMan probe for the non-structural protein NS1 gene were designed. The detection limit was 1 × 102 DNA copies/μL, and the assay was linear in the range of 1 × 102 to 1 × 109 copies/μL. There was no cross-reaction with porcine circovirus 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), classical swine fever virus (CSFV), or Japanese encephalitis virus (JEV). The assay was specific and reproducible. In 41 clinical samples, PPV was detected in 32 samples with the real-time PCR assay and in only 11 samples with a conventional PCR assay. The real-time assay using the TaqMan-system can therefore be practically used for studying the epidemiology and management of PPV.

Multi-functional DEAD-box helicase 5 (DDX5), which is important in transcriptional regulation, is hijacked by diverse viruses to facilitate viral replication. However, its regulatory effect in antiviral innate immunity remains unclear. We found that DDX5 interacts with the N6-methyladenosine (m6A) writer METTL3 to regulate methylation of mRNA through affecting the m6A writer METTL3–METTL14 heterodimer complex. Meanwhile, DDX5 promoted the m6A modification and nuclear export of transcripts DHX58, p65, and IKKγ by binding conserved UGCUGCAG element in innate response after viral infection. Stable IKKγ and p65 transcripts underwent YTHDF2-dependent mRNA decay, whereas DHX58 translation was promoted, resulting in inhibited antiviral innate response by DDX5 via blocking the p65 pathway and activating the DHX58-TBK1 pathway after infection with RNA virus. Furthermore, we found that DDX5 suppresses antiviral innate immunity in vivo. Our findings reveal that DDX5 serves as a negative regulator of innate immunity by promoting RNA methylation of antiviral transcripts and consequently facilitating viral propagation. Author summary: DEAD-box helicase 5 (DDX5) greatly contributes to cancer development and facilitation of viral propagation. However, how DDX5 manipulates host cell processes to facilitate replication remains poorly understood. In this study, we found DDX5 is a negative antiviral regulator through manipulating N6-methyladenosine (m6A) of transcripts in innate immunity. Firstly, DDX5 recruited the RNA m6A "writer" METTL3 to control the m6A writer complex, then specifically promoted m6A modification and nuclear export of DDX5 binding transcripts by binding conserved UGCUGCAG element in innate immune response, ultimately, leading to RNA decay of antiviral transcripts in a YTHDF2-dependent manner. Consequently, DDX5 played vital roles in cellular RNA metabolisms to negatively regulate innate immune response to viral infection. It is the first time to unravel DDX5 as an important component that mediates modification of N6-methyladenosine of mRNA in regulating innate immunity. [ABSTRACT FROM AUTHOR]