Induction Of Receptor Research Articles

FADD-deficient mouse embryonic fibroblasts undergo RIPK1-dependent apoptosis and autophagy after NB-UVB irradiation

Sun or therapy-related ultraviolet B (UVB) irradiation induces different cell death modalities such as apoptosis, necrosis/necroptosis and autophagy. Understanding of mechanisms implicated in regulation and execution of cell death program is imperative for prevention and treatment of skin diseases. An essential component of death-inducing complex is Fas-associated protein with death domain (FADD), involved in conduction of death signals of different death modalities. The purpose of this study was to enlighten the role of FADD in the selection of cell death mode after narrow-band UVB (NB-UVB) irradiation using specific cell death inhibitors (carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]- fluoromethylketone (zVAD-fmk), Necrostatin-1 and 3-Methyladenine) and FADD-deficient (FADD−/−) mouse embryonic fibroblasts (MEFs) and their wild type (wt) counterparts. The results imply that lack of FADD sensitized MEFs to induction of receptor–interacting protein 1 (RIPK1)-dependent apoptosis by the generation of reactive oxygen species (ROS), but without activation of the proteins p53, Bax and Bcl-2 as well as without the enrolment of calpain-2. Autophagy was established as a contributing factor to NB-UVB-induced death execution. By contrast, wt cells triggered intrinsic apoptotic pathway that was resistant to the inhibition by zVAD-fmk and Necrostatin-1 pointing to the mechanism overcoming the cell survival. These findings support the role of FADD in prevention of autophagy-dependent apoptosis.

Hippocampal activation of 5-HT1B receptors and BDNF production by vagus nerve stimulation in rats under chronic restraint stress.

A growing body of evidence shows that the electrical stimulation of the vagus nerve can improve mental illness including depression. Here, we investigated whether the vagus nerve stimulation (VNS) is involved in regulating the responsiveness of hippocampal neurons in rats under chronic restraint stress (CRS). c-Fos protein signals were detected 2hr after VNS in 5-HT1A receptor-positive neurons in the dorsal raphe nucleus (DRN) as well as in the nucleus tractus solitarius (NTS). Chronic VNS was performed on a daily basis for 2weeks using an implanted microelectrode in rats that had undergone CRS for 2weeks. We found that the levels of both 5-HT1B receptors and phospho-Erk1/2 were decreased in parallel in the hippocampal neurons of CRS animals and then increased to the baseline levels by chronic VNS. Hippocampal induction of 5-HT1B receptors and phospho-Erk1/2 by VNS was diminished after the injection of 5,7-dihydroxytryptamine (5,7-DHT), a neurotoxin of serotonergic neurons, into the DRN. Hippocampal production of brain-derived neurotrophic factor (BDNF) was also upregulated by VNS, but the treatment of 5,7-DHT abrogated the effects of VNS on BDNF induction. VNS in CRS animals improved the behavioral scores in forced swimming test (FST) compared to sham-stimulated control. Our results suggest that VNS-mediated serotonergic input via 5-HT1B receptors into the hippocampal neurons may activate BDNF pathway and improve depressive-like behaviors in CRS animals.

Contribution of Adrenal Glands to Intratumor Androgens and Growth of Castration-Resistant Prostate Cancer.

Tumor androgens in castration-resistant prostate cancer (CRPC) reflect de novo intratumoral synthesis or adrenal androgens. We used C.B.-17 SCID mice in which we observed adrenal CYP17A activity to isolate the impact of adrenal steroids on CRPC tumors in vivo. We evaluated tumor growth and androgens in LuCaP35CR and LuCaP96CR xenografts in response to adrenalectomy (ADX). We assessed protein expression of key steroidogenic enzymes in 185 CRPC metastases from 42 patients. Adrenal glands of intact and castrated mice expressed CYP17A. Serum DHEA, androstenedione (AED), and testosterone (T) in castrated mice became undetectable after ADX (all P < 0.05). ADX prolonged median survival (days) in both CRPC models (33 vs. 179; 25 vs. 301) and suppressed tumor steroids versus castration alone (T 0.64 pg/mg vs. 0.03 pg/mg; DHT 2.3 pg/mg vs. 0.23 pg/mg; and T 0.81 pg/mg vs. 0.03 pg/mg, DHT 1.3 pg/mg vs. 0.04 pg/mg; all P ≤ 0.001). A subset of tumors recurred with increased steroid levels, and/or induction of androgen receptor (AR), truncated AR variants, and glucocorticoid receptor (GR). Metastases from 19 of 35 patients with AR positive tumors concurrently expressed enzymes for adrenal androgen utilization and nine expressed enzymes for de novo steroidogenesis (HSD3B1, CYP17A, AKR1C3, and HSD17B3). Mice are appropriate for evaluating adrenal impact of steroidogenesis inhibitors. A subset of ADX-resistant CRPC tumors demonstrate de novo androgen synthesis. Tumor growth and androgens were suppressed more strongly by surgical ADX than prior studies using abiraterone, suggesting reduction in adrenally-derived androgens beyond that achieved by abiraterone may have clinical benefit. Proof-of-concept studies with agents capable of achieving true "nonsurgical ADX" are warranted.

Hepatitis C virus infection, genotypes and mechanism of insulin resistance

Background: Hepatitis C virus (HCV) infection is a major global public health problem that causes profound metabolic abnormalities, primarily in insulin-sensitive target tissues, notably the phenomenon of steatosis or fatty liver. The route of transmission and genetic mutation of HCV, together with the lack of reliable nation-specific epidemiological data on the distribution of genotypes and sub-genotypes of this RNA virus, provide significant challenges to correct diagnosis and effective treatment. Issue: HCV-induced insulin resistance in HCV-infected individuals is independent of the occurrence of metabolic syndrome and diabetes mellitus, primarily type 2 diabetes mellitus, that causes insulin resistance. Some but not all HCV genotypes exert a steatotic effect. However, the molecular mechanism(s) by which HCV infection causes insulin resistance in insulin target tissues or hepatic steatosis is not elucidated clearly. Findings: Mechanisms proposed by experimental studies include interference with insulin signaling pathways, upregulation of genes controlling gluconeogenesis, phosphorylation of insulin receptor substrate proteins, and induction and overexpression of inflammatory cytokines that interact closely with host lipid metabolism. Conclusion: We review HCV genotypes and subtypes, the mechanisms by which HCV infection induces insulin resistance, the virus genotypes and subtypes that are implicated in this and those that are steatotic. We conclude by discussing proposed mechanisms of steatosis and considering HCV laboratory investigation methods from traditional to current techniques.

Sonic hedgehog pathway activation is associated with cetuximab resistance and EPHB3 receptor induction in colorectal cancer.

A major problem of colorectal cancer (CRC) targeted therapies is relapse caused by drug resistance. In most cases of CRC, patients develop resistance to anticancer drugs. Cetuximab does not show many of the side effects of other anticancer drugs and improves the survival of patients with metastatic CRC. However, the molecular mechanism of cetuximab resistance is not fully understood.Methods: EPHB3-mediated cetuximab resistance was confirmed by in vitro western blotting, colony-forming assays, WST-1 colorimetric assay, and in vivo xenograft models (n = 7 per group). RNA-seq analysis and receptor tyrosine kinase assays were performed to identify the cetuximab resistance mechanism of EPHB3. All statistical tests were two-sided.Results: The expression of EFNB3, which upregulates the EPHB3 receptor, was shown to be increased via microarray analysis. When resistance to cetuximab was acquired, EPHB3 protein levels increased. Hedgehog signaling, cancer stemness, and epithelial-mesenchymal transition signaling proteins were also increased in the cetuximab-resistant human colon cancer cell line SW48R. Despite cells acquiring resistance to cetuximab, STAT3 was still responsive to EGF and cetuximab treatment. Moreover, inhibition of EPHB3 was associated with decreased STAT3 activity. Co-immunoprecipitation confirmed that EGFR and EPHB3 bind to each other and this binding increases upon resistance acquisition, suggesting that STAT3 is activated by the binding between EGFR and EPHB3. Protein levels of GLI-1, SOX2, and Vimentin, which are affected by STAT3, also increased. Similar results were obtained in samples from patients with CRC.Conclusion: EPHB3 expression is associated with anticancer drug resistance.

Botanical Formulation HX109 Ameliorates TP-Induced Benign Prostate Hyperplasia in Rat Model and Inhibits Androgen Receptor Signaling by Upregulating Ca2+/CaMKKβ and ATF3 in LNCaP Cells.

Benign prostatic hyperplasia (BPH) is a common disease in the elderly male population throughout the world. Among other factors, androgen dysregulation has been known to play major roles in its pathogenesis. HX109 is a botanical formulation prepared from a mixture of Taraxacum officinale, Cuscuta australis, and Nelumbo nucifera, which have traditionally been used—usually along with other plants—to treat urinary diseases. An ethanol extract was prepared from a mixture of these three plants, and its quality was controlled through cell-based bioassays and by quantification of several marker compounds by high-performance liquid chromatography (HPLC). In the testosterone propionate (TP)-induced prostate hyperplasia rat model, oral administration of HX109 ameliorated prostate enlargement and histological changes induced by TP. In LNCaP cells, a human prostate epithelial cell line, HX109 repressed AR-mediated cell proliferation and the induction of androgen receptor (AR) target genes at the transcriptional level without affecting the translocation or expression of AR. Such effects of HX109 on AR signaling were mediated through the control of activating transcriptional factor 3 (ATF3) expression, phosphorylation of calcium/calmodulin-dependent protein kinase kinase β (CaMKKβ), and increases in intracellular calcium, as evidenced by data from experiments involving ATF3-specific siRNA, CaMKKβ inhibitor, and calcium chelator, respectively. Taken together, our data suggest that HX109 might be used as a starting point for developing therapeutic agents for the treatment of BPH.

Adaptive NK Cells Are Expanded By NKG2C-Agonist Antibodies and Retain Their Cytotoxic and Secretory Properties

Natural killer (NK) cells can eliminate myeloid leukemia cells after stem cell transplant, yet many patients relapse. Several reports have described significant decreases in relapse rates post-transplant in recipients that experienced cytomegalovirus (CMV) reactivation. Our group has shown that reduced relapse rates correlated with expansion of adaptive NK cells, which can be defined by surface expression of the maturation marker CD57 and the activating receptor NKG2C. Adaptive NK cells live longer, are more resistant to immune suppression than canonical NK cells, exhibit properties of immune memory, and their presence in peripheral blood is only observed in CMV seropositive individuals. Given the in vitro and in vivo evidence for enhanced function of adaptive NK cells against malignant cells, we sought to develop a method to preferentially expand this subset in vitro. To this end, we stimulated NK cells isolated from the peripheral blood of healthy CMV seropositive donors for 7 days with IL-15 and either a plate-bound control IgG isotype antibody or an antibody that was determined to be a strong agonist of NKG2C signaling in redirected antibody-dependent cellular cytotoxicity (ADCC) assays. We observed robust proliferation of NKG2C+ NK cells when stimulated with the anti-NKG2C agonist antibody (Figure 1A). High proliferation rates were accompanied by significant induction of the checkpoint inhibitory receptors programmed death-1 (PD-1) and lymphocyte activation gene-3 (LAG-3) (Figure 1B), with LAG-3 expression nearly doubling on NKG2C+ NK cells. LAG-3 is a unique checkpoint inhibitor that ligates MHC class II molecules and is highly expressed on lymphocytes in patients with various types of malignancy.The stimulated NK cells were also highly active, as evidenced by a complete switch from a CD45RA+CD45RO- to CD45RA-CD45RO+ phenotype (Figure 1C). CD45ROexpression on lymphocytes has been associated with better prognosis in solid tumors including melanoma, lung, and endometrial cancers suggesting a potent ability to target malignant cells.Despite the increased expression of checkpoint inhibitory receptors, expanded NKG2C+ NK cells remained highly functional. In assays where NK cells were used as effectors against K562 cells, (a myeloid leukemia cell line), we found that NKG2C agonist antibody-expanded NK cells exhibited higher frequencies of tumor necrosis factor (TNF) and interferon (IFN)-γ production compared to NK cells from control IgG isotype cultures. Surprisingly, we found that NK cells expressing LAG-3, instead of exhibiting functional exhaustion, were highly responsive to tumor targets (Figure 1D). To directly test the cytotoxic function of adaptive NK cells expanded with the anti-NKG2C antibody, we analyzed NK cell-mediated killing of dye-labeled K562 cells over the course of 48 hours by live imaging using an IncuCyte Zoom instrument. We observed more rapid killing kinetics and more robust overall target cell killing by adaptive NK cells expanded with the anti-NKG2C agonist antibody relative to the IgG isotype control condition (Figure 1E).Our data show that CD57+NKG2C+, adaptive NK cells can be efficiently expanded using NKG2C-agonist antibodies. These cells exhibit potent killing abilities and enhanced cytokine secretion, making them ideal candidates for adoptive cell transfer in hematologic malignancies. We also show that LAG-3, a checkpoint inhibitory receptor, is actually associated with enhanced NK cell function in these in vitro assays. This is likely because of the relatively short period of activation (7 days) to which the NK cells are exposed during expansion. There are at least four LAG-3 targeted therapies currently in clinical trials with more in development. A better understanding of the mechanisms of expression of this important receptor and its impact on NK cell function will aid in the rational use of checkpoint inhibitory receptor therapies for cancer patients. [Display omitted] DisclosuresHouchins:Bio-Techne Inc: Employment. Cichocki:Fate Therapeutics Inc.: Consultancy, Research Funding.

Endothelial Microsomal Prostaglandin E Synthetase-1 Upregulates Vascularity and Endothelial Interleukin-1β in Deteriorative Progression of Experimental Autoimmune Encephalomyelitis.

Microsomal prostaglandin E synthetase-1 (mPGES-1) is an inducible terminal enzyme for the production of prostaglandin E2 (PGE2). In experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis, mPGES-1 is induced in vascular endothelial cells (VECs) around inflammatory foci and facilitates inflammation, demyelination, and paralysis. Therefore, we investigated the role of CD31-positive VECs in mPGES-1-mediated EAE aggravation using immunohistochemical analysis and imaging of wild-type (wt) and mPGES-1-deficient (mPGES-1−/−) mice. We demonstrated that EAE induction facilitated vascularity in inflammatory lesions in the spinal cord, and this was significantly higher in wt mice than in mPGES-1−/− mice. In addition, endothelial interleukin-1β (IL-1β) production was significantly higher in wt mice than in mPGES-1−/− mice. Moreover, endothelial PGE2 receptors (E-prostanoid (EP) receptors EP1–4) were expressed after EAE induction, and IL-1β was induced in EP receptor-positive VECs. Furthermore, IL-1 receptor 1 expression on VECs was increased upon EAE induction. Thus, increased vascularity is one mechanism involved in EAE aggravation induced by mPGES-1. Furthermore, mPGES-1 facilitated the autocrine function of VECs upon EP receptor induction and IL-1β production, modulating mPGES-1 induction in EAE.

Endoplasmic Reticulum Stress Activated by Androgen Enhances Apoptosis of Granulosa Cells via Induction of Death Receptor 5 in PCOS.

Polycystic ovary syndrome (PCOS) is associated with hyperandrogenism and growth arrest of antral follicles. Previously, we found that endoplasmic reticulum (ER) stress is activated in granulosa cells of antral follicles in PCOS, evidenced by activation of unfolded protein response (UPR) genes. Based on this observation, we hypothesized that ER stress is activated by androgens in granulosa cells of antral follicles, and that activated ER stress promotes apoptosis via induction of the UPR transcription factor C/EBP homologous protein (CHOP) and subsequent activation of death receptor (DR) 5. In this study, we found that testosterone induced expression of various UPR genes, including CHOP, as well as DR5, in cultured human granulosa-lutein cells (GLCs). Pretreatment with the ER stress inhibitor tauroursodeoxycholic acid (TUDCA) inhibited testosterone-induced apoptosis and expression of DR5 and CHOP. Knockdown of CHOP inhibited testosterone-induced DR5 expression and apoptosis, and knockdown of DR5 inhibited testosterone-induced apoptosis. Pretreatment with flutamide, as well as knockdown of androgen receptor, decreased testosterone-induced DR5 and CHOP expression, as well as apoptosis. Expression of DR5 and CHOP was upregulated in GLCs obtained from patients with PCOS, as well as in granulosa cells of antral follicles in ovarian sections obtained from patients with PCOS and dehydroepiandrosterone-induced PCOS mice. Treatment of PCOS mice with TUDCA decreased apoptosis and DR5 expression in granulosa cells of antral follicles, with a concomitant reduction in CHOP expression. Taken together, our findings indicate that ER stress activated by hyperandrogenism in PCOS promotes apoptosis of granulosa cells of antral follicles via induction of DR5.

Vildagliptin protects endothelial cells against high glucose-induced damage.

Hyperglycemia or high blood sugar is one of the major pathological characteristics of diabetes. The endothelium is the inner layer of the vascular wall and is directly exposed to various stimuli in blood vessels. As a characteristic of diabetes, chronic high glucose is known to be harmful to endothelial cells. In this study, we found that vildagliptin, an available type 2 diabetes agent, protects primary human aortic endothelial cells (HAECs) against damage induced by high glucose. Our data indicate that vildagliptin improves the decrease in endothelial viability and reduces the release of lactate dehydrogenase (LDH) induced by high glucose. Vildagliptin potently suppresses high glucose-induced generation of reactive oxygen species (ROS) and production of vascular inflammatory factors including tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8), intercellular cell adhesion molecule-1 (ICAM-1) and monocyte chemotactic protein 1 (MCP-1). Moreover, vildagliptin suppresses adhesion of monocytes to endothelial cells and induction of toll-like receptor 4 (TLR-4) in HAECs caused by high glucose. Mechanistically, we found that the cellular protective effects mediated by vildagliptin involve suppression of nuclear factor-kappa B (NF-κB) nuclear signals. Collectively, our data indicate that vildagliptin possesses a protective function in vascular cells.

A6066 Induction of (pro)renin receptor by hypoxia in trophoblasts and its possible role in hypertensive disorders of pregnancy

A6066 Induction of (pro)renin receptor by hypoxia in trophoblasts and its possible role in hypertensive disorders of pregnancy

Immunotoxicity of titanium dioxide nanoparticles via simultaneous induction of apoptosis and multiple toll-like receptors signaling through ROS-dependent SAPK/JNK and p38 MAPK activation.

BackgroundTitanium dioxide nanoparticles (TiO2 NPs) represent a scientific breakthrough in the areas of biological and medicinal applications. Interaction of TiO2 NPs with components of innate immune system remains elusive.AimThis study explored in vitro immunotoxicity of murine macrophage RAW 264.7 to TiO2 NPs (20 nm, negative charge) and its underlying molecular mechanism by way of immunoredox profiling.Materials and methodsIn this study, chemically synthesized BSA-functionalized TiO2 NPs (20 nm, negative charge) were characterized and immunotoxicity was investigated on RAW 264.7 cells.ResultsWe found that reactive oxygen species levels significantly increased with increasing nitric oxide production, whereas depleting endogenous antioxidant super oxide dismutase as well as nuclear factor erythroid 2-related factor 2 (Nrf2) protein levels. Furthermore, NPs exposure increased the expression of apoptotic factors such as BAX, BIM, and PUMA with disruption of mitochondrial membrane potential (Δψm) that lead to decrease in immunocytes. Molecular immune profiling revealed the activation of multiple toll-like receptors (TLRs) 4/9/12/13 simultaneously with the phosphorylation of p-p38MAPK and p-SAPK/c-Jun N-terminal kinase (JNK) compared to untreated control.ConclusionCollectively, this study shows that the molecular nature of TiO2SA20(−) NP-induced immunotoxicity in RAW 264.7 macrophage is simultaneous induction of immunocyte apoptosis and multiple TLRs signaling through oxidative stress-dependent SAPK/JNK and p38 mitogen-associated protein kinase activation. This is the first study to address newer molecular mechanism of TiO2SA20(−) NP-induced immunotoxicity.

Disruption of oocyte maturation by selected environmental chemicals in zebrafish

Oocyte maturation can be a target of endocrine disruption by environmental chemicals capable of acting as hormone mimics, receptor blockers, and/or enzyme inhibitors. Six environmental chemicals (genistein, endosulfan, malathion, iprodione, carbaryl, and glyphosate) were selected to determine their ability to interfere with oocyte maturation in zebrafish. The translucent oocytes undergoing germinal vesicle (nucleus) breakdown (GVBD) were counted and expressed as a ratio of oocytes undergoing GVBD and total oocytes exposed. The GVBD increased significantly in oocytes exposed to 10 IU/ml to 100 IU/ml human chorionic gonadotropin (hCG). The lowest effective concentration of genistein that inhibited hCG-induced GVBD was 30 μM, while endosulfan inhibited it at 0.03 μM concentration. In addition, malathion inhibited hCG-induced GVBD at the lowest concentration of 60 μM. These inhibitory effects were likely due to the chemicals acting as estrogen mimics, induction of estrogen receptors, or increase in aromatase activity resulting in enhanced estrogen action. Fungicide iprodione, possibly acting as a progestin mimic, promoted hCG-induced GVBD at the lowest concentration of 20 μM, while the weed killer glyphosate inhibited hCG-induced GVBD starting at the 50 μM concentration. These results demonstrate the feasibility of using fully grown zebrafish oocytes arrested at the prophase I stage in an in vitro incubation system to evaluate the effects of a variety of environmental chemicals on oocyte maturation.

DOX-Vit D, a Novel Doxorubicin Delivery Approach, Inhibits Human Osteosarcoma Cell Proliferation by Inducing Apoptosis While Inhibiting Akt and mTOR Signaling Pathways.

Doxorubicin (DOX) is a very potent and effective anticancer agent. However, the effectiveness of DOX in osteosarcoma is usually limited by the acquired drug resistance. Recently, Vitamin D (Vit-D) was shown to suppress the growth of many human cancer cells. Taken together, we synthesized DOX-Vit D by conjugating Vit-D to DOX in order to increase the delivery of DOX into cancer cells and mitigate the chemoresistance associated with DOX. For this purpose, MG63 cells were treated with 10 µM DOX or DOX-Vit D for 24 h. Thereafter, MTT, real-time PCR and western blot analysis were used to determine cell proliferation, genes and proteins expression, respectively. Our results showed that DOX-Vit D, but not DOX, significantly elicited an apoptotic signal in MG63 cells as evidenced by induction of death receptor, Caspase-3 and BCLxs genes. Mechanistically, the DOX-Vit D-induced apoptogens were credited to the activation of p-JNK and p-p38 signaling pathway and the inhibition of proliferative proteins, p-Akt and p-mTOR. Our findings propose that DOX-Vit D suppressed the growth of MG63 cells by inducing apoptosis while inhibiting cell survival and proliferative signaling pathways. DOX-Vit D may serve as a novel drug delivery approach to potentiate the delivery of DOX into cancer cells.

Retraction: ROS and CHOP Are Critical for Dibenzylideneacetone to Sensitize Tumor Cells to TRAIL through Induction of Death Receptors and Downregulation of Cell Survival Proteins.

Retraction: ROS and CHOP Are Critical for Dibenzylideneacetone to Sensitize Tumor Cells to TRAIL through Induction of Death Receptors and Downregulation of Cell Survival Proteins.

Editor's Note: Zerumbone Enhances TRAIL-Induced Apoptosis through the Induction of Death Receptors in Human Colon Cancer Cells: Evidence for an Essential Role of Reactive Oxygen Species.

Editor's Note: Zerumbone Enhances TRAIL-Induced Apoptosis through the Induction of Death Receptors in Human Colon Cancer Cells: Evidence for an Essential Role of Reactive Oxygen Species.

Effects of irradiation with narrowband-ultraviolet B on up-regulation of histamine H1 receptor mRNA and induction of apoptosis in HeLa cells and nasal mucosa of rats

Narrowband-ultraviolet B (NB-UVB) phototherapy is used for the treatment of atopic dermatitis. Previously, we reported that irradiation with 200 mJ/cm2 of 310 nm NB-UVB suppressed phorbol-12-myristate-13-acetate (PMA)-induced up-regulation of histamine H1 receptor (H1R) gene expression without induction of apoptosis in HeLa cells. However, the effect of NB-UVB irradiation on nasal symptoms is still unclear. Here, we show that low dose irradiation with 310 nm NB-UVB alleviates nasal symptoms in toluene 2,4-diisocyanate (TDI)-sensitized allergy model rats. Irradiation with 310 nm NB-UVB suppressed PMA-induced H1R mRNA up-regulation in HeLa cells dose-dependently at doses of 75–200 mJ/cm2 and reversibly at a dose of 150 mJ/cm2 without induction of apoptosis. While, at doses of more than 200 mJ/cm2, irradiation with 310 nm NB-UVB induced apoptosis. Western blot analysis showed that the suppressive effect of NB-UVB irradiation on H1R gene expression was through the inhibition of ERK phosphorylation. In TDI-sensitized rat, intranasal irradiation with 310 nm NB-UVB at an estimated dose of 100 mJ/cm2 once a day for three days suppressed TDI-induced sneezes and up-regulation of H1R mRNA in nasal mucosa without induction of apoptosis. These findings suggest that repeated intranasal irradiation with low dose of NB-UVB could be clinically used as phototherapy of AR.

CCPG1, an unconventional cargo receptor for ER-phagy, maintains pancreatic acinar cell health

ER stress-mediated induction of a new vertebrate-specific autophagy cargo receptor, CCPG1 (cell-cycle progression gene 1), drives degradation of endoplasmic reticulum. CCPG1 acts via ATG8-family interaction and, non-canonically, via discrete interactions with FIP200. CCPG1 ameliorates ER stress in the exocrine pancreas. This has potential implications for inflammation and cancer, discussed here.

Chrysin Ameliorates Malfunction of Retinoid Visual Cycle through Blocking Activation of AGE-RAGE-ER Stress in Glucose-Stimulated Retinal Pigment Epithelial Cells and Diabetic Eyes.

Diabetes-associated visual cycle impairment has been implicated in diabetic retinopathy, and chronic hyperglycemia causes detrimental effects on visual function. Chrysin, a naturally occurring flavonoid found in various herbs, has anti-inflammatory, antioxidant, and neuroprotective properties. The goal of the current study was to identify the retinoprotective role of chrysin in maintaining robust retinoid visual cycle-related components. The in vitro study employed human retinal pigment epithelial (RPE) cells exposed to 33 mM of glucose or advanced glycation end products (AGEs) in the presence of 1–20 μM chrysin for three days. In the in vivo study, 10 mg/kg of chrysin was orally administrated to db/db mice. Treating chrysin reversed the glucose-induced production of vascular endothelial growth factor, insulin-like growth factor-1, and pigment epithelium-derived factor (PEDF) in RPE cells. The outer nuclear layer thickness of chrysin-exposed retina was enhanced. The oral gavage of chrysin augmented the levels of the visual cycle enzymes of RPE65, lecithin retinol acyltransferase (LRAT), retinol dehydrogenase 5 (RDH5), and rhodopsin diminished in db/db mouse retina. The diabetic tissue levels of the retinoid binding proteins and the receptor of the cellular retinol-binding protein, cellular retinaldehyde-binding protein-1, interphotoreceptor retinoid-binding protein and stimulated by retinoic acid 6 were restored to those of normal mouse retina. The presence of chrysin demoted AGE secretion and AGE receptor (RAGE) induction in glucose-exposed RPE cells and diabetic eyes. Chrysin inhibited the reduction of PEDF, RPE 65, LRAT, and RDH5 in 100 μg/mL of AGE-bovine serum albumin-exposed RPE cells. The treatment of RPE cells with chrysin reduced the activation of endoplasmic reticulum (ER) stress. Chrysin inhibited the impairment of the retinoid visual cycle through blocking ER stress via the AGE-RAGE activation in glucose-stimulated RPE cells and diabetic eyes. This is the first study demonstrating the protective effects of chrysin on the diabetes-associated malfunctioned visual cycle.

Effects of multi and selective targeted tyrosine kinase inhibitors on function and signaling of different bladder cancer cells

BackgroundSignaling of receptor tyrosine kinases (RTK) is dysregulated in various malignancies including bladder cancer. RTKs trigger pro-proliferative, anti-apoptotic and metastatic signaling pathways. Here, we assessed the effects of a selective tyrosine kinase inhibitor (TKI) (BGJ398) targeting fibroblast growth factor receptor (FGFR) and a pan-TKI (TKI258) targeting (FGFR), platelet derived growth factor receptor (PDGFR) and vascular endothelial growth factor receptor (VEGFR) in bladder cancer cells. MethodsLevels of mRNA transcripts were measured in nine human cell lines by quantitative RT-PCR. Cell function was assessed for viability, colony formation, migration, apoptosis and proliferation. Protein mediators of signal transduction were measured by Western-blot. ResultsmRNA transcripts encoding RTK-related components, transcription factors, epithelial and mesenchymal transition (EMT) markers as well as cell cycle and apoptotic factors were determined in the cell lines. Principal component analysis ordered one epithelial-like cell cluster (5637, BFTC-905, MGHU4, RT112) and one mesenchymal-like cell cluster (T24, UMUC3, HU456, TCC-SUP). Cell response scores towards TKI258 and BGJ398 treatment were heterogeneous between cell lines and correlated with certain transcript levels. Analysis of signal transduction pathways revealed inhibition of fibroblast growth factor receptor (FGFR) signaling and induction of cell cycle dependent kinase (CDKN1A, p21) in epithelial-like cells differing in this regard from responses to mesenchymal-like cells that exhibited inhibition of mitogen-activated protein kinase (MAPK). ConclusionRTK and EMT related transcript analysis separate bladder cancer cells in two clusters. Functional responses towards TKI258 and BGJ398 treatment of bladder Fcancer cells were heterogeneous with deviating effects on signaling and possibly different therapeutic outcome.