Quantification of Central and Eastern China's atmospheric CH4 enhancement changes and its contributions based on machine learning approach.

Methane is the second largest anthropogenic greenhouse gas, and changes in atmospheric methane concentrations can reflect the dynamic balance between its emissions and sinks. Therefore, the monitoring of CH4 concentration changes and the assessment of underlying driving factors can provide scientific basis for the government's policy making and evaluation. China is the world's largest emitter of anthropogenic methane. However, due to the lack of ground-based observation sites, little work has been done on the spatial-temporal variations for the past decades and influencing factors in China, especially for areas with high anthropogenic emissions as Central and Eastern China. Here to quantify atmospheric CH4 enhancements trends and its driving factors in Central and Eastern China, we combined the most up-to-date TROPOMI satellite-based column CH4 (xCH4) concentration from 2018 to 2022, anthropogenic and natural emissions, and a random forest-based machine learning approach, to simulate atmospheric xCH4 enhancements from 2001 to 2018. The results showed that (1) the random forest model was able to accurately establish the relationship between emission sources and xCH4 enhancement with a correlation coefficient (R²) of 0.89 and a root mean-square error (RMSE) of 11.98 ppb; (2)The xCH4 enhancement only increased from 48.21±2.02 ppb to 49.79±1.87 ppb from the year of 2001 to 2018, with a relative change of 3.27%±0.13%; (3) The simulation results showed that the energy activities and waste treatment were the main contributors to the increase in xCH4 enhancement, contributing 68.00% and 31.21%, respectively, and the decrease of animal ruminants contributed -6.70% of its enhancement trend.

Early Pregnancy Regulates Expression of IkappaB Family in Ovine Spleen and Lymph Nodes.

Early pregnancy modulates the maternal immune system, including the spleen and lymph nodes, which participate in maternal innate and adaptive immune responses. Methods: Ovine spleens and lymph nodes were sampled at day 16 of the estrous cycle, and at days 13, 16 and 25 of gestation, and qRT-PCR, Western blot and immunohistochemistry analysis were used to analyze the expression of the IκB family, including BCL-3, IκBα, IκBß, IκBε, IKKγ, IκBNS and IκBζ. Early pregnancy induced expression of BCL-3, IκBα, IκBε, IKKγ and IκBζ, and expression of BCL-3, IκBß and IκBNS peaked at day 16 of pregnancy in the spleen. However, early pregnancy suppressed the expression of BCL-3 and IκBNS, but stimulated the expression of IκBß and IκBζ, and expression levels of IκBα, IκBß, IκBε and IKKγ peaked in lymph nodes at days 13 and/or 16 of pregnancy. Early pregnancy changed the expression of the IκB family in the maternal spleen and lymph node in a tissue-specific manner, suggesting that the modulation of the IκB family may be involved in regulation of maternal functions of the spleen and lymph nodes, which are necessary for the establishment of maternal immune tolerance during early pregnancy in sheep.

Expression of nuclear factor kappa B in ovine maternal inguinal lymph nodes during early pregnancy.

BACKGROUND: Pregnancy-induced immunological changes contribute to the maternal immune tolerance. Nuclear factor kappa B (NF-κB) pathway participates in regulating both innate and adaptive immunities, and lymph nodes play key roles in adaptive immune reaction. However, it is unclear whether early pregnancy changes the expression of NF-κB family in maternal lymph node in sheep. METHODS: In this study, the samples of inguinal lymph nodes were collected from ewes on day 16 of the estrous cycle, and on days 13, 16 and 25 of pregnancy, and expression of NF-κB family, including NF-κB p105 (NFKB1), NF-κB p100 (NFKB2), p65 (RELA), RelB (RELB) and c-Rel (REL), were analyzed through real-time quantitative PCR, Western blot and immunohistochemical analysis. RESULTS: The expression levels of NF-κB p105 and c-Rel downregulated, but NF-κB p100 upregulated on day 25 of pregnancy. The expression levels of p65, RelB and c-Rel peaked at day 13 of pregnancy, and expression level of RelB was higher during early pregnancy comparing to day 16 of the estrous cycle. In addition, p65 protein was located in the subcapsular sinus and lymph sinuses. CONCLUSION: This paper reported for the first time that early pregnancy has effects on the expression of NF-κB family, which may contribute to the maternal immunoregulation through blood circulation and lymph circulation during early pregnancy in sheep.

Expression of melatonin receptors and CD4 in the ovine thymus, lymph node, spleen and liver during early pregnancy.

As a pineal gland hormone, melatonin acts through its receptors to modulate the immune system. The immune system is composed of primary and secondary organs, and immune organs are adapted to the presence of the fetal alloantigen during pregnancy. However, it is unclear whether melatonin affects maternal immune organs during early pregnancy in sheep. In this study, the ovine thymus, lymph node, spleen and liver were sampled at day 16 of the oestrous cycle, and at days 13, 16 and 25 of pregnancy. The expression of melatonin receptor 1A (MT1), melatonin receptor 1B (MT2) and cluster of differentiation 4 (CD4) was detected by quantitative real-time polymerase chain reaction, Western blot and immunohistochemistry experiments. Our results showed that during early pregnancy there was an upregulation of MT1 mRNA and protein in the thymus, lymph node and liver, and there was a downregulation in the spleen. The expression of MT2 mRNA and protein was increased in the thymus but decreased in the spleen and liver, and there was no significant change in the lymph node during early pregnancy. CD4 protein was upregulated in the thymus, lymph node and liver, but there were no significant changes in the spleen during early pregnancy. In conclusion, early pregnancy induces tissue-specific expression of MT1, MT2 and CD4, which may be due to the different functions of the thymus, lymph node, spleen and liver. Further, melatonin is involved in immune regulation of the maternal thymus, lymph node, spleen and liver during early pregnancy in sheep.

Expression profiles of interferon-stimulated gene 15 and prostaglandin synthases in the ovine lymph nodes during early pregnancy.

Lymph nodes are distributed all over the body and are part of the lymphatic system. The interferon-stimulated gene 15 kDa protein (ISG15) and prostaglandins (PGs) are involved in the establishment of pregnancy and are expressed in the uterus during early pregnancy in sheep. In this study, the ovine lymph nodes were obtained on Day 16 of the estrous cycle, and Days 13, 16, and 25 of pregnancy, and the expression of ISG15 and PG synthases, including cyclooxygenase 1 (COX-1), COX-2, prostaglandin E (PGE) synthase (PTGES), and a PGF synthase (aldo-keto reductase family 1, member B1, AKR1B1) were detected by quantitative real-time polymerase chain reaction, western blot analysis, and immunohistochemistry analysis. Our results showed that there were peaks in the expression of mRNAs and the proteins of ISG15, COX-1, COX-2, PTGES, and AKR1B1 in the lymph nodes during early pregnancy and that the COX-2 and AKR1B1 proteins were limited to the subcapsular sinus and lymph sinuses. In conclusion, the ISG15, COX-1, COX-2, PTGES, and AKR1B1 were upregulated in the maternal lymph nodes, which may be beneficial for the development of conceptus, maternal systemic immunoregulation, and anti-luteolysis during early pregnancy in sheep.

Immunostimulatory CpG on Carbon Nanotubes Selectively Inhibits Migration of Brain Tumor Cells.

Even when treated with aggressive current therapies, patients with glioblastoma usually survive less than two years and exhibit a high rate of recurrence. CpG is an oligonucleotide that activates the innate immune system via Toll-like receptor 9 (TLR9) activation. Injection of CpG into glioblastoma tumors showed promise as an immunotherapy in mouse models but proved disappointing in human trials. One aspect of glioma that is not addressed by CpG therapy alone is the highly invasive nature of glioma cells, which is associated with resistance to radiation and chemotherapy. Here, we demonstrate that single-walled carbon nanotubes noncovalently functionalized with CpG (SWNT/CpG), which retain the immunostimulatory property of the CpG, selectively inhibit the migration of glioma cells and not macrophages without affecting cell viability or proliferation. SWNT/CpG also selectively decreased NF-κB activation in glioma cells, while activating macrophages by induction of the TLR9/NF-κB pathway, as we have previously reported. The migration inhibition of glioma cells was correlated with selective reduction of intracellular levels of reactive oxygen species (ROS), suggesting that an antioxidant-based mechanism mediates the observed effects. To the best of our knowledge, SWNT/CpG is the first nanomaterial that inhibits the migration of cancer cells while stimulating the immune system.

Expression of genes associated with luteolysis in peripheral blood mononuclear cells during early pregnancy in cattle.

The conceptus-derived signals that initiate maternal recognition of pregnancy act primarily on the endometrium to inhibit the development of luteolysis, thus modifying the expression of genes in the corpus luteum. The involvement of peripheral blood mononuclear cells (PBMCs) in the formation of this anti-luteolytic mechanism during early pregnancy is uncertain. In this study, PBMCs from non-pregnant and early-pregnant cows were sampled to explore the expression of genes associated with luteolysis, including AKR1B1 (aldo-keto reductase family 1, member B1; a bovine prostaglandin F synthase), PTGFR (PGF2α receptor), OXT (oxytocin), PTGES (PGE synthase), PTGER1 (PGE2 receptor 1), and PGR (progesterone receptor). OXT and PTGFR transcript abundance was low in PBMCs at Day 18 in pregnant individuals. PGR and PTGER1 mRNA abundance was significantly higher at Day 30 in pregnant individuals. AKR1B1 and PTGES transcript abundance was significantly higher at Day 18 in PBMCs from non-pregnant individuals, yet AKR1B1 and PTGES protein abundance was elevated at Day 30 in pregnant individuals-although AKR1B1 dimer may be significantly higher at Day 18 in non-pregnant PBMCs. In conclusion, changes in bovine PBMC gene expression are associated with luteolysis during early pregnancy, which implicate the influence of circulating blood components in controlling luteolysis. Mol. Reprod. Dev. 83: 509-515, 2016. © 2016 Wiley Periodicals, Inc.

Maternal nutrient metabolism in the liver during pregnancy.

The liver plays pivotal roles in nutrient metabolism, and correct hepatic adaptations are required in maternal nutrient metabolism during pregnancy. In this review, hepatic nutrient metabolism, including glucose metabolism, lipid and cholesterol metabolism, and protein and amino acid metabolism, is first addressed. In addition, recent progress on maternal hepatic adaptations in nutrient metabolism during pregnancy is discussed. Finally, the factors that regulate hepatic nutrient metabolism during pregnancy are highlighted, and the factors include follicle-stimulating hormone, estrogen, progesterone, insulin-like growth factor 1, prostaglandins fibroblast growth factor 21, serotonin, growth hormone, adrenocorticotropic hormone, prolactin, thyroid stimulating hormone, melatonin, adrenal hormone, leptin, glucagon-like peptide-1, insulin glucagon and thyroid hormone. Our vision is that more attention should be paid to liver nutrient metabolism during pregnancy, which will be helpful for utilizing nutrient appropriately and efficiently, and avoiding liver diseases during pregnancy.

Effects of bisphenol A and tauroursodeoxycholic acid on maturation of porcine oocytes and parthenogenetic development of embryos.

Prolonged exposure of bisphenol A (BPA) has adverse effects on in vitro maturation (IVM) of oocytes, but treatment with tauroursodeoxycholic acid (TUDCA) can improve the IVM and development of embryos. The purpose of this study was to investigate the effects of BPA and both BPA and TUDCA on IVM and parthenogenetic development of embryos. The results showed that BPA treatment adverse effects on the cumulus expansion index, survival rate, polar body rate, mitochondrial distribution of the oocytes after maturation culture, and that it also decreased the cleavage rate and blastocyst rate of embryos after parthenogenetic develpoment. In addition, BPA treatment upregulated expression of genes related to endoplasmic reticulum stress and apoptosis and increased the intracellular reactive oxygen species (ROS) level, while it decreased expression of genes related to cumulus expansion. However, the supplementation of TUDCA relieved these adverse effects of BPA except polar body rate, blastocyst rate, and expression of BCL2 and PTGS1. In conclusion, the supplementation of TUDCA can partly attenuate the negative effects of BPA on IVM and parthenogenetic development of embryos, possibly by modification of the expression of genes related to endoplasmic reticulum stress, apoptosis and cumulus expansion, intracellular ROS level, and mitochondrial distribution.

Effects of early pregnancy on NOD-like receptor expression in the ovine endometrium.

Introduction: Nucleotide-binding domain (NOD)-like receptors (NLRs) are expressed in the endometrium, and involved in modulating the female innate immune responses. There are conceptus-endometrial interactions during pregnancy, which ensure immune homeostasis of the maternal-fetal interface. The purpose of this study was to explore the effects of early pregnancy on NLR expression in the ovine endometrium. Methods: Endometrial tissues were collected at day 16 of the estrous cycle, and at days 13, 16 and 25 of pregnancy (n = 6 for each group), and RT-qPCR, western blot and immunohistochemistry analysis were used to analyze the expression of NLRs, including NOD1, NOD2, major histocompatibility complex class II transactivator (CIITA), neuronal apoptosis inhibitor protein (NAIP), NLR family, pyrin domain-containing 1 (NLRP1), NLRP3 and NLRP7. Results: Expression levels of NOD1, NOD2, NAIP, CIITA, NLRP1 and NLRP3 declined, but expression level of NLRP7 increased in the endometria during early pregnancy compared with nonpregnant ewes. In addition, NOD2 and CIITA proteins were located in the endometrium in a protein type-, cell type- and pregnancy status-specific manner. Discussion: Early pregnancy modulated expression of NLR family in the ovine endometrium, which may be essential for conceptus-endometrial interactions and maternal-fetal interface immune homeostasis.

Pregnancy influences expression of interferon-stimulated genes, progesterone receptor and progesterone-induced blocking factor in ovine thyroid.

OBJECTIVE: Embryonic interferon-tau (IFNT) and progesterone affect expression of interferonstimulated genes (ISGs), progesterone receptor (PGR) and progesterone-induced blocking factor (PIBF) in the ovine thyroid. METHODS: Thyroids of ewes were sampled at day 16 of nonpregnancy, days 13, 16, and 25 of pregnancy, and real-time quantitative polymerase chain reaction assay, western blot and immunohistochemistry were used to detect expression of ISGs, PGR, and PIBF. RESULTS: Free ISG15 protein was undetected, but ISG15 conjugated proteins upregulated at day 16 of pregnancy, and expression levels of ISG15 conjugated proteins, PGR isoform (70 kDa), PIBF, interferon-gamma-inducible protein 10 and myxovirusresistance protein 1 peaked, but expression level of signal transducer and activator of transcription 1 was the lowest at day 16 of pregnancy. In addition, the expression levels of PGR isoform (70 kDa) and signal transducer and activator of transcription 1 (STAT1) decreased, but levels of PGR isoform (43 kDa), 2',5'-oligoadenylate synthetase, IP-10 and MX1 increased at day 25 of pregnancy comparing with day 16 of the estrous cycle. CONCLUSION: Early pregnancy affects expression of ISGs, PGR, and PIBF in maternal thyroid through IFNT and progesterone, which may regulate thyroid autoimmunity and thyroid hormone secretion in ewes.

Modulation of PI3K/AKT/mTOR signaling pathway in the ovine liver and duodenum during early pregnancy.

The liver and intestine play a critical role in nutrient absorption, storage, and metabolism. The aim of this study was to evaluate expression pattern of phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of the rapamycin (mTOR) signaling pathway that included PI3K, AKT1, mTOR, FoxO1, SREBP-1, PPARα, PTEN and FXR in the maternal liver and duodenum. Ovine livers and duodenums were sampled at day 16 of the estrous cycle, and at days 13, 16 and 25 of gestation, and RT-qPCR, western blot and immunohistochemistry analysis were used to detect mRNA and protein expression. The results showed that expression of PI3K, AKT1, p-mTOR, FoxO1, SREBP-1 and PTEN upregulated in the maternal liver, and PPARα upregulated in the duodenum. However, expression of FoxO1, SREBP-1 and PTEN in the duodenum downregulated during early pregnancy. In addition, expression levels of SREBP-1, PTEN and PPARα in the maternal liver, and PI3K in the duodenum peaked at day 13 of pregnancy. In addition, expression levels of PI3K, p-mTOR and FoxO1 in the liver, and AKT1 and p-mTOR in the duodenum peaked at day 16 of pregnancy. Nevertheless, expression levels of FXR both in the maternal liver duodenum downregulated at days 13 and 16 of pregnancy. In conclusion, early pregnancy regulated expression pattern of PI3K/AKT/mTOR signaling pathway in the ovine liver and duodenum in a pregnancy stage-specific and tissue-specific manner, which may be necessary for the adaptations in maternal hepatic nutrient metabolism and intestinal nutrient absorption early pregnancy.

The impact of dynamic traffic and wind conditions on green infrastructure performance to improve local air quality.

Road traffic represents the dominant source of air pollution in urban street canyons. Local wind conditions greatly impacts the dispersion of these pollutants, yet street trees complicate ventilation in such settings. This case study adopts a novel modelling framework to account for dynamic traffic and wind conditions to identify the optimal street tree configuration that prevents a deterioration in air quality. Measurement data from a shallow to moderately deep street canyon (average 0.5 H/W aspect ratio and four lanes of 1-way traffic) in Dublin, Ireland was used for model calibration. The computational fluid dynamics (CFD) models were used to examine scenarios of dynamic traffic flows within each traffic lane with respect to its impact on local PM2.5 concentrations on adjacent footpaths, segmenting air quality monitoring results based on different wind conditions for model calibration. The monitoring campaign identified higher PM2.5 concentrations on the leeward (north) footpath, with average differences of 14.1 % (2.15 µg/m3) for early evening peaks. The modelling results demonstrated how street trees negatively impacted air quality on the windward footpath in parallel wind conditions regardless of leaf area density (LAD) or tree spacing, with mixed results observed on the leeward footpath in varying traffic flows and wind speeds. Perpendicular wind direction models and high wind speed exacerbated poor air quality on the windward footpath for all tree spacing models, while improving the air quality on the leeward footpath. The findings advise against planting high-LAD trees in this type of street, with a minimum of 20 m spacing for low-LAD trees to balance reducing local air pollution and ventilation capacity in the street. This study highlights the complexities of those in key decision-marking roles and demonstrates the need to adopt a transparent framework to ensure adequate modelling evidence can inform tree planting in city streets.

Lapsed El Niño impact on Atlantic and Northwest Pacific tropical cyclone activity in 2023.

A typical El Niño event often results in suppressed tropical cyclone (TC) genesis frequency (TCGF) over the North Atlantic (NA) and a distinct northwest-southeast dipole pattern in TCGF anomaly over the western North Pacific (WNP). The 2023 saw a strong El Niño event but surprisingly active NA and suppressed WNP TC activities. Here, we present that these unprecedented deviations were driven by the record-warm NA, a record-breaking negative phase of the Pacific Meridional Mode (PMM), and background global warming. Results from high-resolution global model experiments demonstrate that extraordinary Atlantic warming dominated the increased NA TCGF and contributed equally with the PMM to the suppressed WNP TCGF, overshadowing El Niño's impact. Global warming also contributed to the observed TCGF anomalies. Our findings demonstrate that the typical influence of strong El Niño events on regional TC activity could be markedly altered by other climate modes, highlighting the complexity of TC genesis in a warming world.

Regulation of IkappaB Protein Expression by Early Gestation in the Thymus of Ewes.

The thymus is an essential component of maternal immune systems that play key roles in recognizing the placenta as immunologically foreign. The inhibitor of the NF-κB (IκB) family has essential effects on the NF-κB pathway; however, it is unclear whether early pregnancy modulates the expression of the IκB family in the thymus. In this study, maternal thymuses were sampled on day 16 of nonpregnancy and different gestation stages in the ovine, and the expression of IκB proteins was analyzed. The data showed that B cell leukemia-3 and IκBß increased; however, IκBα, IκBε, and IKKγ deceased during gestation. Furthermore, there was an increase in IκBNS and IκBζ expression values on day 13 of pregnancy; however, this decreased on day 25 of gestation. In summary, the expression of the IκB family was modulated in the thymus during early gestation, suggesting that the maternal thymus can be associated with maternal immunologic tolerance and pregnancy establishment in ewes.

Maternal hepatic immunology during pregnancy.

The liver plays pivotal roles in immunologic responses, and correct hepatic adaptations in maternal immunology are required during pregnancy. In this review, we focus on anatomical and immunological maternal hepatic adaptations during pregnancy, including our recent reports in this area. Moreover, we summarize maternal pregnancy-associated liver diseases, including hyperemesis gravidarum; intrahepatic cholestasis of pregnancy; preeclampsia, specifically hemolysis, elevated liver enzymes, and low platelet count syndrome; and acute fatty liver of pregnancy. In addition, the latest information about the factors that regulate hepatic immunology during pregnancy are reviewed for the first time, including human chorionic gonadotropin, estrogen, progesterone, growth hormone, insulin like growth factor 1, oxytocin, adrenocorticotropic hormone, adrenal hormone, prolactin, melatonin and prostaglandins. In summary, the latest progress on maternal hepatic anatomy and immunological adaptations, maternal pregnancy-associated diseases and the factors that regulate hepatic immunology during pregnancy are discussed, which may be used to prevent embryo loss and abortion, as well as pregnancy-associated liver diseases.

Expression of IkappaB Family in the Ovine Liver during Early Pregnancy.

During normal pregnancy, there is a dynamic regulation of the maternal immune system, including the liver, to accommodate the presence of the allogeneic foetus in the uterus. However, it was unclear that the expression of the IkappaB (IκB) family was regulated in the ovine maternal liver during early pregnancy. In this study, sheep livers were collected at day 16 of the oestrous cycle (NP16), and days 13, 16 and 25 of gestation (DP13, DP16 and DP25), and RT-qPCR, Western blot and immunohistochemistry analysis were used to analyse the expression of the IκB family, including B cell leukemia-3 (BCL-3), IκBα, IκBß, IκBε, IKKγ, IκBNS and IκBζ. The results revealed that expression of BCL-3, IκBß, IκBε and IKKγ peaked at DP16, and the expression of IκBα was increased during early pregnancy. In addition, the expression of IκBζ peaked at DP13 and DP16, and IκBNS peaked at DP13. IκBß and IKKγ proteins were located in the endothelial cells of the proper hepatic arteries and portal veins, and hepatocytes. In conclusion, early pregnancy changed the expression of the IκB family, suggesting that the modulation of the IκB family may be related to the regulation of maternal hepatic functions, which may be favourable for pregnancy establishment in sheep.

Changes in expression levels of Nod-like receptors in the spleen of ewes.

Nucleotide-binding oligomerization domain receptors (NOD-like receptors, NLRs) have critical effects on interfaces of the immune and reproductive systems, and the spleen plays a key role in both innate and adaptive immune functions. It is hypothesized that NLR family participates in maternal splenic immune regulation during early pregnancy in sheep. In this study, maternal spleens were collected on day 16 of the estrous cycle, and days 13, 16 and 25 of gestation (n = 6 for each group) in ewes. Expression of NLR family, including NOD1, NOD2, class II transactivator (CIITA), NLR family apoptosis inhibitory protein (NAIP), nucleotide-binding oligomerization domain, Leucine rich repeat and Pyrin domain containing 1 (NLRP1), NLRP3 and NLRP7, was analyzed using quantitative real-time PCR, Western blot and immunohistochemistry analysis. The results revealed that expression levels of NOD1, NOD2, CIITA and NLRP3 were downregulated at days 13 and 16 of pregnancy, but expression of NLRP3 was increased at day 25 of pregnancy. In addition, expression values of NAIP and NLRP7 mRNA and proteins were improved at days 16 and 25 of pregnancy, and NLRP1 was peaked at days 13 and 16 of pregnancy in the maternal spleen. Furthermore, NOD2 and NLRP7 proteins were limited to the capsule, trabeculae and splenic cords. In summary, early pregnancy changes expression of NLR family in the maternal spleen, which may be related with the maternal splenic immunomodulation during early pregnancy in sheep.

RAGE ablation attenuates glioma progression and enhances tumor immune responses by suppressing galectin-3 expression.

BACKGROUND: Malignant gliomas consist of heterogeneous cellular components that have adopted multiple overlapping escape mechanisms that overcome both targeted and immune-based therapies. The receptor for advanced glycation end products (RAGE) is a member of the immunoglobulin superfamily that is activated by diverse proinflammatory ligands present in the tumor microenvironment. Activation of RAGE by its ligands stimulates multiple signaling pathways that are important in tumor growth and invasion. However, treatment strategies that only target the interaction of RAGE with its ligands are ineffective as cancer therapies due to the abundance and diversity of exogenous RAGE ligands in gliomas. METHODS: As an alternative approach to RAGE ligand inhibition, we evaluated the genetic ablation of RAGE on the tumorigenicity of 2 syngeneic murine glioma models. RAGE expression was inhibited in the GL261 and K-Luc gliomas by shRNA and CRSPR/Cas9 techniques prior to intracranial implantation. Tumor growth, invasion, and inflammatory responses were examined by histology, survival, Nanostring, and flow cytometry. RESULTS: Intracellular RAGE ablation abrogated glioma growth and invasion by suppressing AKT and ERK1/2 activities and by downregulating MMP9 expression. Interestingly, RAGE inhibition in both glioma models enhanced tumor inflammatory responses by downregulating the expression of galectin-3 and potentiated immunotherapy responses to immune checkpoint blockade. CONCLUSIONS: We demonstrated that intracellular RAGE ablation suppresses multiple cellular pathways that are important in glioma progression, invasion, and immune escape. These findings strongly support the development of RAGE ablation as a treatment strategy for malignant gliomas.

Differential expression pattern of ISG15 in different tissue explants and cells induced by various interferons.

Interferon stimulated gene 15 (ISG15), an ubiquitin cross-reactive protein, can conjugate to target proteins. Unlike ubiquitination, protein modification by ISG15 does not target protein for degradation, but enhances the cellular response to interferon (IFN), which plays a key role in antiviral responses. In this study, Western blot and/or immunocytochemistry were performed to explore the ISG15 expression patterns in explants of bovine endometrium, mammary gland and kidney, as well as Madin-Darby bovine kidney (MDBK), endometrial and mammary cells stimulated by IFN-α, -ß, and -τ. Western blot indicated that there are differential minimum antiviral units among recombinant bovine interferon-α (rbIFN-α, 10(2) IU/mL), rbIFN-ß (10(3) IU/mL) and rbIFN-τ (10(4) IU/mL) in regard to stimulating saturation expression of free and ISG15-conjugated proteins by MDBK cells and endometrial and mammary explants. These results were further confirmed through immunocytochemical analysis of MDBK, endometrial and mammary cells. For the first time it has been shown that the expression pattern of ISG15-conjugated proteins occurs in a tissue-specific manner. Furthermore, the present findings provide the first evidence of 10- to 100-fold differences in minimum antiviral units of rbIFN-α, rbIFN-ß, and rbIFN-τ in regard to stimulating saturation expression of ISG15.