Antiviral and synergistic effects of photo-energy with acyclovir on herpes simplex virus type 1 infection.

This experimental study aimed to evaluate the antiviral and synergistic effects of photoenergy irradiation on human herpes simplex virus type I (HSV-1) infection. We assessed viral replication, plaque formation, and relevant viral gene expression to examine the antiviral and synergistic effects of blue light (BL) with acyclovir treatment. Our results showed that daily BL (10 J/cm2) irradiation inhibited plaque-forming ability and decreased viral copy numbers in HSV-1-infected monkey kidney epithelial Vero cells and primary human oral keratinocyte (HOK) cells. Combined treatment with the antiviral agent acyclovir and BL irradiation increased anti-viral activity, reducing viral titers and copy numbers. In particular, accumulated BL irradiation suppressed characteristic viral genes including UL19 and US6, and viral DNA replication-essential genes including UL9, UL30, UL42, and UL52 in HOK cells. Our results suggest that BL irradiation has anti-viral and synergistic properties, making it a promising therapeutic candidate for suppressing viral infections in clinical trials.

Blue Light Inhibits Proliferation of Metastatic Cancer Cells by Regulating Translational Initiation: A Synergistic Property with Anticancer Drugs.

The aim of this study was to examine the inhibitory effect of blue light (BL) on the proliferation of metastatic cancer cells and synergistic properties with chemo-drugs. BL significantly inhibited the proliferation of B cell lymphoma (A20 and RAMOS) cells in a dose-dependent manner. Anti-proliferative effect of BL irradiation was identified to be associated with the inhibition of proliferating-cell nuclear antigen expression and cell cycle by decreasing S-phase cells. Consistent with its inhibitory effects, BL irradiation at 20 J/cm2 daily for 10 days inhibited metastasis of cancer cells which were distributed and invaded to other organs including bone marrow, liver, kidney, etc., and induced paraplegia, thereby leading to an increased survival rate of tumor-bearing mice. Anti-proliferative activity of BL was expanded in solid tumor cells including pancreatic carcinoma (Mia PaCa-2, PANC-1), lung carcinoma A549 and colorectal carcinoma HCT116 cells. Additionally, combination with chemo-drugs such as 5-FU and gemcitabine resulted in an increase in the anti-proliferative activity after BL irradiation accompanied by regulating mRNA translational process via inhibition of p70S6K, 4EBP-1 and eIF4E phosphorylation during cellular proliferation. These results indicate the anti-metastatic and photo-biogoverning abilities of BL irradiation as a potent therapeutic potential for repressing the progression of tumor cells.

Blue light irradiation exerts anti-viral and anti-inflammatory properties against herpes simplex virus type 1 infection.

The aim of this study was to investigate the antiviral and anti-inflammatory functions of blue light (BL) in cutaneous viral infections. Previously, we examined the photo-biogoverning role of 450 nm BL in SARS-CoV-2-infected cells, which showed that photo-energy could inhibit viral activation depending on the number of photons. However, the communication network between photo-energy irradiation and immune cells involved in viral infections has not been clarified. We verified viral activation, inflammatory responses, and relevant downstream cascades caused by human simplex virus type I (HSV-1) after BL irradiation. To examine the antiviral effect of BL, we further tested whether BL could disturb viral absorption or entry into host cells. The results showed that BL irradiation, but not green light (GL) exposure, specifically decreased plaque-forming activity and viral copy numbers in HSV-1-infected cells. Accumulated BL irradiation inhibited the localization of viral proteins and the RNA expression of characteristic viral genes such as UL19, UL27, and US6, thus exerting to an anti-viral effect. The results also showed that BL exposure during viral absorption interfered with viral entry or destroyed the virus, as assessed by plaque formation and quantitative PCR assays. The levels of the pro-inflammatory mediators interleukin (IL)-18 and IL-1ß in M1-polarized macrophages were increased by HSV-1 infection. However, these increases were attenuated by BL irradiation. Importantly, BL irradiation decreased cGAS and STING expression, as well as downstream NF-κB p65, in M1-polarized HSV-1-infected macrophages, demonstrating anti-viral and anti-inflammatory properties. These findings suggest that BL could serve as an anti-viral and anti-inflammatory therapeutic candidate to treat HSV-1 infections.

Evaluation of Photobiogoverning Role of Blue Light Irradiation on Viral Replication.

Most recently, severe acute respiratory syndrome coronavirus-2 has triggered a global pandemic without successful therapeutics. The goal of the present study was to define the antiviral effect and therapeutic action of blue light irradiation in SARS-CoV-2-infected cells. Vero cells were infected with SARS-CoV-2 (NCCP43326) or mock inoculum at 50 pfu/well. After blue light irradiation, the inhibitory effect was assessed by qPCR and plaque reduction assay. When Vero cells were irradiated to blue light ranging from 1.6 to 10 J cm-2 , SARS-CoV-2 replication was inhibited by up to 80%. The antiviral effect of blue light irradiation was associated with translation suppression via the phosphorylation of eIF2α by prolonging endoplasmic reticulum (ER) stress. The levels of LC3A/B and Beclin-1, which are key markers of autophagy, and the levels of PERK and PDI for ER stress were highly increased, whereas caspase-3 cleavage was inhibited after blue light irradiation in the later stage of infection. Our data revealed that blue light irradiation exerted antiviral and photo-biogoverning activities by prolonging ER stress and stimulating autophagy progression during viral infection. The findings increase our understanding of how photo-energy acts on viral progression and have implications for use in therapeutic strategies against COVID-19.

Conditioned media from blue light-emitting diode-exposed fibroblasts have an anti-inflammatory effect in vitro.

We have previously reported the protective effects of blue light-emitting diode (BLED)-stimulated cell metabolites on cell injury. To further examine the effect of conditioned media (CM) derived from BLED (5 J/cm2)-exposed human normal fibroblasts (CMBL5) for clinical application, we have used the choline chloride and phenol red-free media and then concentrated CMBL5 using a centrifugal filter unit. The collected CMBL5-lower part (CMBL5-LO) has evaluated the inflammatory protein expression profile in LPS-stimulated RAW264.7 cells. Comprehensive metabolomic profiling of CMBL5-LO was carried out using hybrid tandem mass spectrometry. Treatment with CMBL5-LO showed the cytoprotective effect on apoptotic cell death, but rather increased apoptotic cells after treatment with CMBL5-upper part (CMBL5-UP). In addition, CMBL5-LO inhibited several chemo-attractants, including interleukin (IL)-6, macrophage inflammatory protein (MIP)-2, chemokine (C-C motif) ligand 5 (CCL5), granulocyte colony-stimulating factor (GCSF), and monocyte chemoattractant protein-1 (MCP-1) expression. Pro-inflammatory nitric oxide was decreased after CMBL5-LO treatment, but not by CMBL5-UP treatment. Interestingly, treatment with CMBL5-LO stimulated expression of heme oxygenase-1, indicating its anti-inflammatory property. Most endoplasmic reticulum (ER) stress proteins except for transcription factor C/EBP homologous protein (CHOP) were highly expressed after irradiation with BLED in cells. Further studies are needed to examine the precise mechanism by CMBL5-LO in cells.

Rodent Leukocyte Isolation and Radiolabeling for Inflammation Imaging Study.

PURPOSE: The objective of this study was to describe to develop methods of rodent leukocyte isolation and radiolabeling for in vivo inflammation imaging. METHODS: Thigh muscle inflammation was induced by injection of collagenase. Blood was collected from the jugular vein and separated by Histopaque. The collected cells were incubated in a 37 °C CO2 incubator for 1~2 h. After incubation, 99mTc-HMPAO and 18F-FDG were used to treat leukocytes followed by incubation for 30 min. 99mTc-HMPAO and 18F-FDG labeled autologous leukocytes were injected into the tail veins of rats. The images were then acquired at various time points. Image-based lesion to normal muscle ratio was compared. RESULTS: After Histopaque separation, the proportion of lymphocytes was higher than that of other cell types. After CO2 incubation, the collected leukocytes were viable, while room temperature exposed leukocytes without CO2 incubation were non-viable. Granulocytes, especially, were more quickly influenced by various conditions than the mononuclear cells. Labeling efficiencies of 99mTc-HMPAO and 18F-FDG were 4.00 ± 2.06 and 1.8%, respectively. 99mTc-HMPAO- and 18F-FDG-labeled leukocytes targeted well the inflamed lesion. 99mTc-HMPAO-labeled leukocytes, but not 18F-FDG-labeled leukocytes, were found in the abdomen activity. CONCLUSION: Inflamed lesions of rats were well visualized using autologous radiolabeled leukocytes. This method might provide good information for understanding inflammatory diseases.

Analysis of Cell Fraction of 99mTc-HMPAO Radiolabeled Leukocytes.

PURPOSE: 99mTc-HMPAO radiolabeled autologous leukocyte scintigraphy is routinely used clinically for infection imaging. Leukocytes are mostly separated via sedimentation. It is unknown whether leukocytes are clearly separated by sedimentation or selectively labeled. Therefore, in this study, the blood cell numbers were investigated after leukocyte radiolabeling to identify the cells strongly radiolabeled by 99mTc-HMPAO. METHODS: This study was performed with leftover blood samples of the patients who underwent 99mTc-HMPAO scintigraphy at Chonbuk National University Hospital (2018-2019). The blood of 22 patients was drawn for 99mTc-HMPAO scintigraphy. WBCs were separated via conventional sedimentation at our clinic and radiolabeled. The concentration of cell components was determined using an automatic hematology analyzer. The cells in the final sample injectate sample were separated using Histopaque and counted with a dose calibrator. RESULTS: The average numbers of RBCs, WBCs, and PLTs in the final injection sample were 79 ± 33, 23.26 ± 11.95, and 229.5 ± 206.57 x 103/µL, respectively. The PLT number was almost 10-fold the number of WBCs. The number of RBCs was nearly 3-fold higher than WBCs [RBC/WBC ratio = 4.67 ± 3.58, and PLT/WBC ratio = 10.65 ± 12.46]. Following Histopaque separation, the activity of each layer showed 99mTc-HMPAO labeling of WBC > RBC > PLT in order. The total activity/cell numbers of WBCs, RBCs and PLTs were 0.016 ± 0.010, 0.005 ± 0.005 and 0.003 ± 0.002, respectively (p > 0.05). CONCLUSION: Although the numbers of RBCs and PLTs were highly increased after sedimentation, their individual cellular activity was lower than that of WBCs. 99mTc-HMPAO was more selective to WBCs than RBCs or PLTs. In conclusion, a higher number of WBCs were radiolabeled compared with RBCs and PLTs.

Characterization of malignant ovarian mass on [18F]FDG PET/CT: using metabolic indices and degree of solidity.

BACKGROUND: The utility of [18F]FDG PET/CT for characterizing malignant ovarian mass has not been extensively studied. Here, we investigated various parameters that could be useful to differentiate malignant ovarian mass. METHODS: We enrolled 51 female patients (53.4±15.0 years), with 86 ovarian masses, who underwent pretreatment [18F]FDG PET/CT. Thirty six lesions were histopathologically confirmed with ovarian serous adenocarcinoma. Thirty one ovarian masses from gastric cancer and 19 masses from colorectal cancer were diagnosed by histopathological study or clinical follow-up. Ovarian masses were evaluated by size, solidity, and metabolic indices. The degree of solidity was scored from 1 to 5 according to the portion of solid and cyst. Metabolic activity was scored to be either positive (≥ liver) or negative (< liver). SUVmax (SUVovary) and the ratio of SUVmax of ovary to SUVmean of the liver (ovary/L ratio) were performed. Age, bilaterality and level of CA 125 were also compared. In statistical analysis, categorical variables were analyzed using Pearson's chi-square test, while continuous variables were evaluated either independent student's t-test or Mann-Whitney Test. Receiveroperating-characteristic analysis was used to obtain optimal cutoff values. RESULTS: Serous adenocarcinoma had significantly higher score in all metabolic indices over metastasis. However, there were no differences in all metabolic indices in ovarian metastasis. In contrast, solidity was different between metastatic mass from gastric and colorectal cancer. Ovarian metastasis from gastric cancer was significantly solid compared with that from colorectal cancer. In comparison of all three masses, solidity and all metabolic indices were significantly different. Patients with serous adenocarcinoma were older and had higher CA-125 level. Between metastases from gastric and colorectal cancer, there were no differences in age, bilaterality and CA-125. CONCLUSIONS: Metabolic indices such as SUVovary and ovary/L ratio could be useful to differentiate serous adenocarcinoma from metastasis. Furthermore, the degree of solidity could play a role in predicting the origin of metastasis.

Protective Effect of BLED-exposed Conditioned Media on Cell Injury.

Previous studies have reported that 450 nm blue light emitting diode (BLED) induces apoptosis through a mitochondria-mediated pathway in cancer cells and reduces the early stage tumor growth. This study was performed to determine the effects of BLED-irradiated cell metabolites on cell injury. Our results showed that conditioned medium (CM) from cells irradiated with low-dose BLED (LCM) inhibited apoptosis and increased cell survival. Cell protection-related proteins were identified in cell metabolites of CM and LCM using 2-DE and MALDI-TOF analysis. Treatment with LCM inhibited apoptotic cell death and increased the live cell population. The cellular protective effect of LCM was associated with keratin and collagen type VI secretion from cells after low dose of BLED irradiation. Interestingly, expression of endoplasmic reticulum stress proteins was dose dependently increased after 4 h BLED irradiation. Only levels of BiP, CHOP and ERO1-Lα were decreased significantly after 24 h incubation, indicating their anti-apoptotic property in these cells. These results indicated that cell metabolites stimulated by low-dose BLED irradiation have a cytoprotective effect on cell injury via increasing transient intracellular ER stress. Further studies remain to provide the molecular mechanisms of LCM for cytoprotective activity.

Inhibitory effect of blue light emitting diode on migration and invasion of cancer cells.

The aim of this study was to determine the effects and molecular mechanism of blue light emitting diode (LED) in tumor cells. A migration and invasion assay for the metastatic behavior of mouse colon cancer CT-26 and human fibrosarcoma HT-1080 cells was performed. Cancer cell migration-related proteins were identified by obtaining a 2-dimensional gel electrophoresis (2-DE) in total cellular protein profile of blue LED-irradiated cancer cells, followed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis of proteins. Protein levels were examined by immunoblotting. Irradiation with blue LED inhibited CT-26 and HT-1080 cell migration and invasion. The anti-metastatic effects of blue LED irradiation were associated with inhibition of matrix metalloproteinase (MMP)-2 and MMP-9 expression. P38 MAPK phosphorylation was increased in blue LED-irradiated CT-26 and HT-1080 cells, but was inhibited after pretreatment with SB203580, a specific inhibitor of p38 MAPK. Inhibition of p38 MAPK phosphorylation by SB203580 treatment increased number of migratory cancer cells in CT-26 and HT-1080 cells, indicating that blue LED irradiation inhibited cancer cell migration via phosphorylation of p38 MAPK. Additionally blue LED irradiation of mice injected with CT-26 cells expressing luciferase decreased early stage lung metastasis compared to untreated control mice. These results indicate that blue LED irradiation inhibits cancer cell migration and invasion in vitro and in vivo.

Data in support of effect of blue LED irradiation in human lymphoma cells.

As a new and preferred light source for phototherapy, blue light emitting diodes (LEDs) with wavelengths of 400-500 nm have been used to treat hyperbilirubinaemia in infantile jaundice [1]. Recent studies report that blue LED irradiation induces apoptosis by stimulating a mitochondrial pathway and reduces the early growth rate of melanoma cells in mice [2]. Here, we detected the induction of apoptotic cell death and formation of autophagosome in human B lymphoma cells after irradiation with blue LED. This paper provides data in support of the research article entitled "Blue light emitting diode induces apoptosis in lymphoid cells by stimulating autophagy" [3].

Blue light emitting diode induces apoptosis in lymphoid cells by stimulating autophagy.

The present study was performed to examine the induction of apoptotic cell death and autophagy by blue LED irradiation, and the contribution of autophagy to apoptosis in B cell lymphoma A20 and RAMOS cells exposed to blue LED. Irradiation with blue LED reduced cell viability and induced apoptotic cell death, as indicated by exposure of phosphatidylserine on the plasma outside membrane and fragmentation of DNA. Furthermore, the mitochondrial membrane potential increased, and apoptotic proteins (PARP, caspase 3, Bax, and bcl-2) were observed. In addition, the level of intracellular superoxide anion (O2(-)) gradually increased. Interestingly the formation of autophagosomes and level of LC3-II were increased in blue LED-irradiated A20 and RAMOS cells, but inhibited after pretreatment with 3-methyladenine (3-MA), widely used as an autophagy inhibitor. Inhibition of the autophagic process by pretreatment with 3-MA blocked blue LED irradiation-induced caspase-3 activation. Moreover, a significant reduction of both the early and late phases of apoptosis after transfection with ATG5 and beclin 1 siRNAs was shown by the annexin V/PI staining, indicating a crucial role of autophagy in blue LED-induced apoptosis in cells. Additionally, the survival rate of mice irradiated with blue LED after injection with A20 cells increased compared to the control group. Our data demonstrate that blue LED irradiation induces apoptosis via the mitochondrial-mediated pathway, in conjunction with autophagy. Further studies are needed to elucidate the precise mechanism of blue LED-induced immune cell death.

Improving Cerebral Blood Flow Through Liposomal Delivery of Angiogenic Peptides: Potential of ¹8F-FDG PET Imaging in Ischemic Stroke Treatment.

UNLABELLED: Strategies to promote angiogenesis can benefit cerebral ischemia. We determined whether liposomal delivery of angiogenic peptides with a known biologic activity of vascular endothelial growth factor benefitted cerebral ischemia. Also, the study examined the potential of (18)F-FDG PET imaging in ischemic stroke treatment. METHODS: Male Sprague-Dawley rats (n = 40) underwent 40 min of middle cerebral artery occlusion. After 15 min of reperfusion, the rats (n = 10) received angiogenic peptides incorporated into liposomes. Animals receiving phosphate-buffered solution or liposomes without peptides served as controls. One week later, (18)F-FDG PET imaging was performed to examine regional changes in glucose utilization in response to the angiogenic therapy. The following day, (99m)Tc-hexamethylpropyleneamine oxime autoradiography was performed to determine changes in cerebral perfusion after angiogenic therapy. Corresponding changes in angiogenic markers, including von Willebrand factor and angiopoietin-1 and -2, were determined by immunostaining and polymerase chain reaction analysis, respectively. RESULTS: A 40-min period of middle cerebral artery occlusion decreased blood perfusion in the ipsilateral ischemic cortex of the brain, compared with that in the contralateral cortex, as measured by (99m)Tc-hexamethylpropyleneamine oxime autoradiography. Liposomal delivery of angiogenic peptides to the ischemic hemisphere of the brain attenuated the cerebral perfusion defect compared with controls. Similarly, vascular density evidenced by von Willebrand factor-positive staining was increased in response to angiogenic therapy, compared with that of controls. This increase was accompanied by an early increase in angiopoietin-2 expression, a gene participating in angiogenesis. (18)F-FDG PET imaging measured at 7 d after treatment revealed that liposomal delivery of angiogenic peptides facilitated glucose utilization in the ipsilateral ischemic cortex of the brain, compared with that in the controls. Furthermore, the change in regional glucose utilization was correlated with the extent of improvement in cerebral perfusion (r = 0.742, P = 0.035). CONCLUSION: Liposomal delivery of angiogenic peptides benefits cerebral ischemia. (18)F-FDG PET imaging holds promise as an indicator of the effectiveness of angiogenic therapy in cerebral ischemia.

Effect of blue light emitting diodes on melanoma cells: involvement of apoptotic signaling.

The present study was undertaken to examine whether blue LED irradiation induces cellular apoptosis in B16-F10 cells and whether it blocks the early growth of melanoma cells in mice. Irradiation with blue LED was observed to reduce cell viability and to induce apoptotic cell death, as accompanied by exposure of phosphatidylserine on the plasma outside membrane and an accumulation of a sub-G1 population. Furthermore, the mitochondrial membrane potential increased, and mitochondria-related apoptotic proteins (cytochrome c, caspase 3, and PARP) were observed. In addition, the level of intracellular superoxide anion (O2(-)) gradually increased. Interestingly the phosphorylation of p53 increased at earlier times under blue LED irradiation, but reduced after exposure for a longer time. Additionally, the thickness of the mice footpad injected with B16-F10 cells decreased significantly until the 9th day of blue LED irradiation, indicating the inhibition of the early growth rate of the melanoma cells. Our data demonstrate that blue LED irradiation induces apoptotic cell death by activating the mitochondria-mediated pathway and reduces the early growth rate of melanoma cells. Further studies are needed to elucidate the precise mechanism of blue LED in melanoma cells.