High toxin concentration in pollen may deter collection by bees in butterfly-pollinated Rhododendron molle.

BACKGROUNDS AND AIMS: The hypothesis that plants evolve features that protect accessible pollen from consumption by flower visitors remains poorly understood. METHODS: To explore potential chemical defenses against pollen consumption, we examined the pollinator assemblage, foraging behaviour, visitation frequency and pollen transfer efficiency in Rhododendron molle, a highly toxic shrub containing Rhodojaponin III. Nutrient (protein and lipid) and toxic components in pollen and other tissues were measured. KEY RESULTS: Overall in the five populations, floral visits by butterflies and bumblebees were relatively more frequent than visits by honeybees. All foraged for nectar but not pollen. Butterflies did not differ from bumblebees in the amount of pollen removed per visit, but deposited more pollen per visit. Pollination experiments indicated that R. molle was self-compatible, but both fruit and seed production were pollen limited. Our analysis indicated that the pollen was not protein-poor and had a higher concentration of the toxic compound Rhodojaponin III than petals and leaves, which compound was undetectable in nectar. CONCLUSION: Pollen toxicity in Rhododendron flowers may discourage pollen robbers (bees) from taking the freely accessible pollen grains, while the toxin-free nectar rewards effective pollinators, promoting pollen transfer. This preliminary study supports the hypothesis that chemical defense in pollen would be likely to evolve in species without physical protection from pollinivores.

FAWPA: A FAW Attack Protection Algorithm Based on the Behavior of Blockchain Miners.

Blockchain has become one of the key techniques for the security of the industrial internet. However, the blockchain is vulnerable to FAW (Fork after Withholding) attacks. To protect the industrial internet from FAW attacks, this paper proposes a novel FAW attack protection algorithm (FAWPA) based on the behavior of blockchain miners. Firstly, FAWPA performs miner data preprocessing based on the behavior of the miners. Then, FAWPA proposes a behavioral reward and punishment mechanism and a credit scoring model to obtain cumulative credit value with the processed data. Moreover, we propose a miner's credit classification mechanism based on fuzzy C-means (FCM), which combines the improved Aquila optimizer (AO) with strong solving ability. That is, FAWPA combines the miner's accumulated credit value and multiple attack features as the basis for classification, and optimizes cluster center selection by simulating Aquila's predation behavior. It can improve the solution update mechanism in different optimization stages. FAWPA can realize the rapid classification of miners' credit levels by improving the speed of identifying malicious miners. To evaluate the protective effect of the target mining pool, FAWPA finally establishes a mining pool and miner revenue model under FAW attack. The simulation results show that FAWPA can thoroughly and efficiently detect malicious miners in the target mining pool. FAWPA also improves the recall rate and precision rate of malicious miner detection, and it improves the cumulative revenue of the target mining pool. The proposed algorithm performs better than ND, RSCM, AWRS, and ICRDS.

Wogonin inhibits in vitro herpes simplex virus type 1 and 2 infection by modulating cellular NF-κB and MAPK pathways.

BACKGROUND: Wogonin, a natural flavonoid-like chemical compound, exhibits anti-inflammatory, antitumor, antiviral, neuroprotective, and anxiolytic effects by modulating a variety of cellular signaling pathways including PI3K-Akt, p53, nuclear factor κB (NF-κB), mitogen-activated protein kinase (MAPK) pathways. In this study, its antiviral effect against herpes simplex virus (HSV) type 1 and 2 (HSV-1 and HSV-2) replication was investigated. RESULTS: Wogonin suppressed HSV-2-induced cytopathic effect (CPE) and reduced viral mRNA transcription, viral protein synthesis, and infectious virion particle titers in a dose-dependent manner. A time-of-drug-addition assay demonstrated that wogonin acted as a postentry viral inhibitor. Wogonin also significantly reduced HSV-induced NF-κB and MAPK pathway activation, which has previously been demonstrated to be important for viral replication. CONCLUSIONS: Our results suggest that the anti-herpes effect of wogonin may be mediated by modulation of cellular NF-κB and JNK/p38 MAPK pathways and imply that wogonin may be useful as an anti-HSV agent.

Herpes simplex virus type 2 infection triggers AP-1 transcription activity through TLR4 signaling in genital epithelial cells.

BACKGROUND: The pattern recognition receptors (PPRs) are the earliest phase of the host defense against pathogens in genital epithelium, and toll-like receptors (TLRs) are best characterized PPRs mediating innate immune responses. Herpes simplex virus type 2 (HSV-2), a member of herpesviridae family, causes one of the most prevalent sexually transmitted infection in the world. In this paper, we described that HSV-2 infection would induce activator protein 1 (AP-1) via TLR4-MyD88/TRIF pathway in human genital epithelial cell. METHODS: TLRs expression profiles and changes was investigated in HSV-2-infected cells. The effect of TLR4-MyD88/TRIF on HSV-2-induced AP-1 activation and viral replication was also evaluated. The TLR4 translocation change was examined after viral infection. Finally, viral ICP0 effect on TLR4 signaling and TLR4-promoter regulation were primarily studied. RESULTS: HSV-2-induced AP-1 activation was dependent on TLR4 and downstream adaptor molecules MyD88 and TRIF. And also, TLR4, MyD88 and TRIF was proved to affect HSV-2 replication. AP-1 activation would also be enhanced via overexpression of myeloid differentiation protein 2 (MD2), implicating that it might be a necessary accessory for TLR4 to sense HSV-2 infection. Protein quantification of cytoplasmic and membrane-associated TLR4 revealed that HSV-2 infection increased membrane-anchoring TLR4 level, but not cytoplasmic ones. Viral ICP0 could augment cellular AP-1, TLR4 promoter activation and TLR4 expression level. The specific inhibitor treatment and transcription factor binding site scanning in TLR4 promoter region showed that AP-1 activity was essential for TLR4-promoter activation. CONCLUSIONS: Taken together, HSV-2 infection could stimulate AP-1 activation via TLR4-MyD88/TRIF axis, and then feedback to up-regulate TLR4 expression in human genital epithelial cells.

Augmented atherogenesis in ApoE-null mice co-exposed to polychlorinated biphenyls and 2,3,7,8-tetrachlorodibenzo-p-dioxin.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and polychlorinated biphenyls (PCBs) are persistent organic pollutants found as complex mixtures in the environment throughout the world. Therefore, humans are ubiquitously and simultaneously exposed to TCDD and PCBs. TCDD and PCBs alone have been linked to atherosclerosis. However, the effects of interactions or synergism between TCDD and PCBs on atherogenesis are unknown. We investigated the possible enhanced atherogenesis by co-exposure to TCDD and PCBs and the potential mechanism(s) involved in this enhancement. Male ApoE(-/-) mice were exposed to TCDD (15 µg/kg) and Aroclor1254 (55 mg/kg, a representative mixture of PCBs) alone or in combination by intraperitoneal injection four times over six weeks of duration. Our results showed that mice exposed to TCDD alone, but not Aroclor1254 alone, developed atherosclerotic lesions. Moreover, we found that atherosclerotic disease was exacerbated to the greatest extent in mice co-exposed to TCDD and Aroclor1254. The enhanced lesions correlated with several pro-atherogenic changes, including a marked increase in the accumulation of the platelet-derived chemokine PF4, and the expression of the proinflammatory cytokine MCP-1 and the critical immunity gene-RIG-I. Our data demonstrated that co-exposure to TCDD and Aroclor1254 markedly enhanced atherogenesis in ApoE(-/-) mice. Significantly, our observations suggest that combined exposure to TCDD and PCBs may be a greater cardiovascular health risk than previously anticipated from individual studies.

A novel highly active AgMOF-based silver single-atom catalyst and its application to the aptamer SERS/RRS for the determination of aflatoxin B1.

A novel highly active silver single-atom catalyst (AgSAC) was prepared by a microwave-assisted solvothermal method using silver covalent organic frameworks (AgMOF) as precursors. It was characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), infrared (IR), and surface-enhanced Raman scattering (SERS). The experiment found that AgSAC has excellent catalytic performance and can heavily catalyze the nano-reaction of chloroauric acid-malic acid (HAuCl4-H2Mi) to generate gold nanoparticles (AuNPs). The produced AuNPs have strong SERS, resonance Rayleigh scattering (RRS) and surface plasmon resonance absorption (Abs) signals. Aflatoxin B1 aptamer (AptAFB1) can be adsorbed to the surface of AgSAC through electrostatic interaction, to reduce the catalytic activity of AgSAC and the SERS/RRS/Abs signal of the system. When the target molecule (AFB1) was added, it will specifically bind to AptAFB1 and release AgSAC, restoring the catalytic activity of AgSAC, thereby restoring the SERS/RRS/Abs signal of the system. Based on this, a simple and sensitive aptamer sensing analysis platform for trace AFB1 was established, and a reasonable catalytic amplification mechanism of AgSAC was proposed. The SERS method exhibited the highest sensitivity, with a linear range of 0.005-0.225 µg/L and a detection limit of 0.002 µg/L.

Characterization of TCHQ-induced genotoxicity and mutagenesis using the pSP189 shuttle vector in mammalian cells.

Tetrachlorohydroquinone (TCHQ) is a major toxic metabolite of the widely used wood preservative, pentachlorophenol (PCP), and it has also been implicated in PCP genotoxicity. However, the underlying mechanisms of genotoxicity and mutagenesis induced by TCHQ remain unclear. In this study, we examined the genotoxicity of TCHQ by using comet assays to detect DNA breakage and formation of TCHQ-DNA adducts. Then, we further verified the levels of mutagenesis by using the pSP189 shuttle vector in A549 human lung carcinoma cells. We demonstrated that TCHQ causes significant genotoxicity by inducing DNA breakage and forming DNA adducts. Additionally, DNA sequence analysis of the TCHQ-induced mutations revealed that 85.36% were single base substitutions, 9.76% were single base insertions, and 4.88% were large fragment deletions. More than 80% of the base substitutions occurred at G:C base pairs, and the mutations were G:C to C:G, G:C to T:A or G:C to A:T transversions and transitions. The most common types of mutations in A549 cells were G:C to A:T (37.14%) and A:T to C:G transitions (14.29%) and G:C to C:G (34.29%) and G:C to T:A (11.43%) transversions. We identified hotspots at nucleotides 129, 141, and 155 in the supF gene of plasmid pSP189. These mutation hotspots accounted for 63% of all single base substitutions. We conclude that TCHQ induces sequence-specific DNA mutations at high frequencies. Therefore, the safety of using this product would be carefully examined.

A stable and sensitive Au metal organic frameworks resonance Rayleigh scattering nanoprobe for detection of SO3 2- in food based on fuchsin addition reaction.

A stable Au metal organic frameworks (AuMOF) nanosol was prepared. It was characterized by electron microscopy and molecular spectral techniques. In pH 6.8 PBS buffer solution, AuMOF nanoprobes exhibit a strong resonance Rayleigh scattering (RRS) peak at 330 nm. After basic fuchsin (BF) adsorbing on the surface of AuMOF, the RRS energy of the nanoprobe donor can be transferred to BF receptor, resulting in a decrease in the RRS intensity at 330 nm. Both sulfite and BF taken place an addition reaction to form a colorless product (SBF) that exhibit weak RRS energy transfer (RRS-ET) between AuMOF and SBF, resulting in the enhancement of the RRS peak. As the concentration of SO3 2-increases, the RRS peak is linearly enhanced. Thus, a new and sensitive RRS-ET method for the detection of SO3 2- (0.160-5.00 µmol/L) was developed accordingly using AuMOF as nanoprobes, with a detection limit of 0.0800 µmol/L. This new RRS method was applied to determination of SO3 2- in food and SO2 in air samples. The recoveries of food and air samples were 97.1-106% and 92.9-106%, and the relative standard deviation (RSD) was 2.10-4.80% and 2.10-4.50%, respectively.

Topology optimization design of three-dimensional multi-material and multi-body structure based on irregular cellular hybrid cellular automata method.

In recent years, Hybrid Cellular Automata Method (HCAM) has been successfully applied to solve structural topology optimization problems. However, there was no report on HCAM research of three-dimensional composite structure composed of multiple materials and multiple bodies. Therefore, in this paper, three-dimensional non-cube cells of irregular size (such as tetrahedral cells with adaptive changes inside length) and Finite Element Method (FEM) are introduced to extend HCAM, which is better and more flexibly to fit complex geometric shapes. Furthermore, a better structure configuration of multi-material and multi-body structure is obtained. The typical example study showed that the proposed topology optimization method could effectively remove the redundant materials of multi-material and multi-body structure, and the optimized structure configuration could still meet the requirements of the original condition after geometric reconstructed. Thus it provided a reference for the intelligent design of other products.

A cholesterol benzoate RRS probe for the determination of trace ammonium ions.

The measurement of NH4+ has attracted considerable attention with the increase of NH4+ emissions in sewage caused by human activities. So far, a variety of photometric and fluorescence methods for the detection of NH4+ have been researched and summarized, but there is no report about the use of liquid crystals (LCs) cholesteryl benzoate (CB) as a resonance Rayleigh scattering (RRS) probe to determine ammonium ions. In the NaAc-HAc buffer solution with pH = 4.80, the yellow compounds 3,5 diacetyl-1,4 dihydrolutidine (DDL) generated by the reaction of NH4+ with acetylacetone (AT) and formaldehyde (HCHO) act as the energy receiver and CB as the donor. Because the RRS spectrum of CB overlaps with the DDL absorption spectrum, resonance Rayleigh scattering energy transfer (RRS-ET) occurs. When the NH4+ concentration increased, the generated DDL increased, and the RRS-ET also increased, so the RRS intensity of the system at 395 nm decreased. For this reason, a fast and sensitive CB RRS-ET method was established to apply to the detection of NH4+ in water. The detection range was 1.00 × 10-3 - 4.66 µg/mL, and the detection limit was 6.62 × 10-3 µg/mL. Using this method to analyze and detect NH4+ in environmental water samples, the precision and recovery rate were between 1.30-9.30% and 95.5-109.9%, respectively. Therefore, this method has the advantages of sensitivity and simplicity.

Troglitazone induced apoptosis via PPARγ activated POX-induced ROS formation in HT29 cells.

OBJECTIVE: In order to investigate the potential mechanisms in troglitazone-induced apoptosis in HT29 cells, the effects of PPARγ and POX-induced ROS were explored. METHODS: [3- (4, 5)-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay, Annexin V and PI staining using FACS, plasmid transfection, ROS formation detected by DCFH staining, RNA interference, RT-PCR & RT-QPCR, and Western blotting analyses were employed to investigate the apoptotic effect of troglitazone and the potential role of PPARγ pathway and POX-induced ROS formation in HT29 cells. RESULTS: Troglitazone was found to inhibit the growth of HT29 cells by induction of apoptosis. During this process, mitochondria related pathways including ROS formation, POX expression and cytochrome c release increased, which were inhibited by pretreatment with GW9662, a specific antagonist of PPARγ. These results illustrated that POX upregulation and ROS formation in apoptosis induced by troglitazone was modulated in PPARγ-dependent pattern. Furthermore, the inhibition of ROS and apoptosis after POX siRNA used in troglitazone-treated HT29 cells indicated that POX be essential in the ROS formation and PPARγ-dependent apoptosis induced by troglitazone. CONCLUSION: The findings from this study showed that troglitazone-induced apoptosis was mediated by POX-induced ROS formation, at least partly, via PPARγ activation.

Sensitive Aptamer SERS and RRS Assays for Trace Oxytetracycline Based on the Catalytic Amplification of CuNCs.

A new method for the determination of oxytetracycline (OTC) has been established by coupling the catalytic amplification reaction of copper nanoclusters (CuNCs) with the aptamer reaction. CuNCs prepared by a wet chemical method have the catalytic activity for the formation of gold nanoparticles (AuNPs) resulting from a HAuCl4-ethanol (En) reaction. The experimental results showed that OTC aptamer (Apt) can be adsorbed on the surface of CuNCs in a non-specific way, thus inhibiting its catalytic activity. When OTC was added to the solution, the OTC-Apt complex was generated by a specific reaction, which made the CuNCs desorb and restore their catalytic activity. With the increase of OTC, the recovery of the catalytic activity of CuNCs is strengthened, the reaction speed is accelerated, and the number of AuNPs is increased. The generated AuNPs exhibited surface enhanced Raman scattering (SERS) signals at 1615 cm-1 in the presence of Vitoria blue 4R (VB4R) molecular probes, and a resonance Rayleigh scattering (RRS) peak at 586 nm. There is a good linear relationship between the intensities of SERS, or RRS, and OTC concentration at the range of 37.5-300 ng/L or 37.5-225 ng/L, respectively. A new SERS and RRS assay for the determination of trace OTC based on the regulation of CuNCs catalysis was established.

Inflammatory response and insulin signaling alteration induced by PCB77.

A specific mechanism whereby inflammation may contribute to cardiovascular diseases (CVD), insulin resistance (IR) and type II diabetes is the induction of endothelial dysfunction placing the vascular endothelium in a key unifying position for the shared pathogenesis of these diseases. However, the mechanisms by which PCBs induce endothelial cell dysfunction are not clearly understood. In the present study, we used human umbilical vascular endothelial cells (HUVEC) as model, and inflammatory response and insulin signaling alteration induced by PCBs were examined. Results showed that PCB77 induced the expression of proinflammatory cytokines including IL-6 and TNFalpha and induced U937 adhesion to HUVEC cells consistent with increased NFkappaB transcription activity. On the other hand, PCB77 blocked insulin-activated Akt signaling pathway, which was restored by pretreatment with TNFalpha neutralization antibody. In conclusion, PCB77 showed the potential to induce the expression of proinflammatory cytokines including IL-6, which has been shown to be powerful independent risk predictor of CVD. And PCB77 was observed to alter insulin-activated Akt signaling by TNFalpha secretion for the first time.

Potential mechanisms involved in ceramide-induced apoptosis in human colon cancer HT29 cells.

OBJECTIVE: To investigate the potential mechanisms of cell death after the treatment with ceramide. METHODS: MTT assay, DNA ladder, reporter assay, FACS and Western blot assay were employed to investigate the potential mechanisms of cell death after the treatment with C2-ceramide. RESULTS: A short-time treatment with C2-ceramide induced cell death, which was associated with p38 MAP kinase activation, but had no links with typical caspase activation or PARP degradation. Rather than caspase inhibitor, Inhibitor of p38 MAP kinase blocked cell death induced by a short-time treatment with ceramide (<12 h). However, inhibition of p38 MAP kinase could not block cell death induced by a prolonged treatment with ceramide (>12 h). Moreover, incubation of cells with ceramide for a long time (>12 h) increased subG1, but reduced S phase accompanied by caspase-dependent and caspase-independent changes including NFkappaB activation. CONCLUSION: Ceramide-induced cell apoptosis involves both caspase-dependent and -independent signaling pathway. Caspase-independent cell death occurring in a relatively early stage, which is mediated via p38 MAP kinase, can progress into a stage involving both caspase-dependent and -independent mechanisms accompanied by cell signaling of MAPKs and NFkappaB.

A Novel Approach for Sericin-Conjugated Silver Nanoparticle Synthesis and Their Potential as Microbicide Candidates.

Silver nanoparticles have been widely applied for biomedical areas owing to their potent antiviral and antibacterial activities. Synthesis of silver nanoparticles using biomacromolecules is more efficient, environment-friendly, and cost-saving compared with the traditional approach. In this paper, a novel approach was developed to establish a reaction system with Ag+-BH4--sericin to synthesize silver nanoparticles conjugated to sericin (AgNPs-Sericin). Sericin could be as a good dispersant and stabilizing agent, which is able to modify nanoscaled AgNPs, the average diameter of which was only 3.78 ± 1.14nm prepared in a 0.3 mg/ml sericin solution. The characterizations of the AgNPs-Sericin were determined by FTIR, thermogravimetry, and XRD analyses. The results showed that the synthesized AgNPs conjugated with sericin as organic phase. Via SAED and XRD analysis, we showed that these AgNPs formed polycrystalline powder with a face-centered cubic structure of bulk metals. Moreover, we investigated the antiviral and antibacterial activities of AgNPs-Sericin, and the results showed that AgNPs-Sericin exhibited potent anti-HIV-1 activity against CCR5-tropic and CXCR4-tropic strains, but no significant cytotoxicity was found toward human genital epithelial cells compared with free silver ions, which are accepted as a commonly used potent antimicrobial agent. Moreover, its antibacterial activity was determined via flow cytometry. The results showed that AgNPs-Sericin could suppress gram-negative (E. coli) and gram-positive (S. aureus) bacteria, but more was potent for the gram-negative one. We concluded that our AgNPs-Sericin could be a potential candidate as a microbicide or antimicrobial agent to prevent sexually transmitted infections.

Quantitative determination of soybean meal content in compound feeds: comparison of near-infrared spectroscopy and real-time PCR.

Standard methods for determining the raw material content of compound feed are little exploited, except for the identification of meat and bone meal in feeds. In this work, near-infrared (NIR) spectroscopy and real-time polymerase chain reaction (PCR) were applied in order to establish new and fast methods for quantification of soybean meal content in compound feeds. The best prediction quality was achieved by using a model based on NIR spectroscopy (R2 = 0.9857, standard error of cross-validation 1.1065). Furthermore, a sensitive qualitative detection method by using the real-time PCR was developed (R2 = 0.976, slope -3.7599). Finally, the differences between the real-time PCR result and the NIR spectroscopy result for a given sample were also treated, and we found that the NIR spectroscopy method provided quite accurate results which approach closely those of the real-time PCR method.

Mechanisms involved in ceramide-induced cell cycle arrest in human hepatocarcinoma cells.

AIM: To investigate the effect of ceramide on the cell cycle in human hepatocarcinoma Bel7402 cells. Possible molecular mechanisms were explored. METHODS: [3- (4, 5)-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay, plasmid transfection, reporter assay, FACS and Western blotting analyses were employed to investigate the effect and the related molecular mechanisms of C2-ceramide on the cell cycle of Bel7402 cells. RESULTS: C2-ceramide was found to inhibit the growth of Bel7402 cells by inducing cell cycle arrest. During the process, the expression of p21 protein increased, while that of cyclinD1, phospho-ERK1/2 and c-myc decreased. Furthermore, the level of CDK7 was downregulated, while the transcriptional activity of PPARgamma was upregulated. Addition of GW9662, which is a PPARgamma specific antagonist, could reserve the modulation action on CDK7. CONCLUSION: Our results support the hypothesis that cell cycle arrest induced by C2-ceramide may be mediated via accumulation of p21 and reduction of cyclinD1 and CDK7, at least partly, through PPARgamma activation. The ERK signaling pathway was involved in this process.

Near-infrared reflectance spectroscopy-based methods for phytase registration in feed industry.

The application of biotechnological products in the feed industry has undergone explosive growth in recent years, and phytase from microorganism accounts for one-third of the entire feed enzyme market. In this study, some differences in the composition of protein and denaturation temperature between two commercial phytases were determined by HPLC and differential scanning calorimetry, which were derived from the same origin of E. coli. At the same time, we found that it was advantageous for near-infrared reflectance spectroscopy (NIRS) to display the protein differences in the commercial phytase, which is most important for ensuring the traceability of biotechnological products in feed and food safety control. Furthermore, NIRS could track the changes in phytase during the spray-drying process and the change of enzyme activity during storage of phytase. Our experiments proved that the information from NIRS could describe well the individual characteristics of the commercial phytase, which indicated that near-infrared reflectance spectra could be exploited to use in the registration system of commercial phytase.

Enterovirus 71 induces autophagy by regulating has-miR-30a expression to promote viral replication.

Enterovirus 71 (EV71), the etiological agent of hand-foot-and-mouth disease, has increasingly become a public health challenge around the world. Previous studies reported that EV71 infection can induce autophagic machinery to enhance viral replication in vitro and in vivo, but did not address the underlying mechanisms. Increasing evidence suggests that autophagy, in a virus-specific manner, may function to degrade viruses or facilitate viral replication. In this study, we reported that EV71 infection of human epidermoid carcinoma (Hep2) and African green monkey kidney cells (Vero) induced autophagy, which is beneficial for viral replication. Our investigation of the mechanisms revealed that EV71 infection resulted in the reduction of cellular miR-30a, which led to the inhibition of Beclin-1, a key autophagy-promoting gene that plays important roles at the early phase of autophagosome formation. We provided further evidence that by modulating cellular miR-30a level through either overexpression or inhibition, one can inhibit or promote EV71 replication, respectively, through regulating autophagic activity.

Harmine blocks herpes simplex virus infection through downregulating cellular NF-κB and MAPK pathways induced by oxidative stress.

Herpes simplex virus types 1 and 2 (HSV-1 and -2) are highly prevalent in many populations and therapeutic options are limited. Both viruses can establish latency by maintaining viral genomes in neurons of sensory ganglia. Primary or recurrent HSV infections may lead to deleterious outcomes: HSV-1 infection may result in corneal blindness and encephalitis and HSV-2 infection leads to herpes genitalis. While no effective vaccine is available, acyclovir is widely used for therapy, which targets and inhibits viral DNA polymerase. Although acyclovir is of low toxicity, resistant strains arise due to persistent use, mainly in immune compromised patients. In our effort to identify new HSV inhibitory molecules, harmine was found to potently inhibit HSV infection. Harmine, a beta-carbon alkaloid with an indole core structure and a pyridine ring, is widely distributed in plants. Earlier studies showed that harmine exhibited pharmacological activities such as antifungal, antimicrobial, antitumor, antiplasmodial and antioxidants. In the current study, we showed that harmine was a potent inhibitor of HSV-2 infection in vitro assays with EC50 value at around 1.47µM and CC50 value at around 337.10µM. The HSV RNA transcription, protein synthesis, and virus titers were reduced by the presence of harmine in a dose dependent manner. Further study on the mechanism of the anti-HSV activity showed that harmine blocked HSV-induced ROS production and the upregulated cytokine/chemokine expression, but our evidence showed that the inhibition of viral replication was unlikely mediated by the blocking of ROS production. We demonstrated that harmine significantly reduced HSV-2-induced NF-κB activation, as well as IκB-α degradation and p65 nuclear translocation. We found that harmine also inhibited HSV-2-mediated p38 kinase and c-Jun N-terminal kinases (JNK) phosphorylation.