Chinese university students' help-seeking behaviors when faced with mental health challenges.

BACKGROUND: Mental illnesses and mental health challenges have become increasingly pervasive among Chinese university students. However, the utilization rate of mental health services is low among students. AIMS: We aimed to explore Chinese university students' help-seeking behaviors to understand how they deal with mental health challenges and use the results to inform the development of effective mental health promotion initiatives. METHODS: In this study, we conducted 13 focus group interviews with students in six universities in Jinan, China, including 91 (62%) female students and 56 (38%) male students. We drew on the Theory of Planned Behaviors to guide our thematic analysis to gain a contextual understanding of participants' accounts on help-seeking. RESULTS: Our results have depicted the help-seeking patterns of Chinese university students and show that there are four major behaviors which are self-reliance, seeking support from peers and families, seeking professional support, and accessing virtual mental health care. CONCLUSION: Results from this study can be used to inform the development of mental health literacy programming for students in universities that share similar contexts, and the study has also opened up a new space for using qualitative approaches to study mental health needs and access to care in diverse populations.

Knowledge of HPV/cervical cancer and acceptability of HPV self-sampling among women living with HIV: A scoping review.

Cervical cancer rates are disproportionately high among women living with the human immunodeficiency virus (wlhiv). Cervical cancer is preventable through hpv screening, regular Pap tests, and early cancer detection. Evidence indicates that hpv and cervical cancer screening are suboptimal among wlhiv, who face a myriad of access barriers. Considering that screening is an effective first-line defense to cervical cancer, we conducted a scoping review with the aim of gaining a better understanding about: (1) the knowledge and perceptions of hpv and cervical cancer screening among wlhiv; and (2) the acceptability of self-sampling for hpv among wlhiv. We searched five electronic databases for peer-reviewed articles that were published in English within the last ten years, reported on studies with hiv-positive women who were aged 16 or older, and satisfied the topics of the review. A total of 621 articles were found. After accounting for duplicates and unmet criteria, 17 articles and 1 abstract, reporting on studies in the United States and Africa, were included in this review. The review highlighted that most wlhiv had inadequate knowledge of hpv transmission and cervical cancer prevention, which influenced their perceptions of risk and susceptibility. Screening barriers included misconceptions about Pap tests, fear of diagnosis of serious illness, perceived pain, embarrassment, bodily modesty, and limited access to female health care providers. This review also affirms that self-sampling is an acceptable and promising screening option for wlhiv. Implications for policy, research, and practice are discussed.

HIV-positive MSM's knowledge of HPV and anal cancer self-sampling: A scoping review.

Human papillomavirus (hpv) infection is the cause of anal squamous cell cancer (ascc) in 80% of cases. Available research has also shown high prevalence of anal hpv infection among men who have sex with men (msm). However, hpv vaccination is low among msm in Canada. In light of this information, we conducted a scoping review with the aim of exploring (1) the knowledge of hpv and anal cancer among hiv-positive msm and (2) the acceptability of hpv and anal cancer self-sampling in this population. In conducting the review, we searched five electronic databases for peer-reviewed articles and abstracts published in English, between 2007 and 2017. A total of 803 articles were retrieved; after accounting for duplicates (n=40) and unmet criteria (n=754), a total of 794 articles were excluded. A final total of nine articles were used in this review. Results of this review show that hiv-positive msm have limited knowledge regarding the risks of anal cancer associated with hiv and hpv coinfection. Furthermore, there is limited research on hpv and anal cancer self-sampling in this population. However, the review of available studies suggested that hiv-positive msm were open to anal cancer self-sampling. It also identified potential barriers to self-sampling. In conclusion, we provide suggestions and future directions for policy-makers and educators to develop inclusive and accessible strategies to reach hiv-positive msm regarding anal cancer education and self-screening.

Observations and impacts of bleach washing on indoor chlorine chemistry.

Ambient levels of chlorinated gases and aerosol components were measured by online chemical ionization and aerosol mass spectrometers after an indoor floor were repeatedly washed with a commercial bleach solution. Gaseous chlorine (Cl2 , 10's of ppbv) and hypochlorous acid (HOCl, 100's of ppbv) arise after floor washing, along with nitryl chloride (ClNO2 ), dichlorine monoxide (Cl2 O), and chloramines (NHCl2 , NCl3 ). Much higher mixing ratios would prevail in a room with lower and more commonly encountered air exchange rates than that observed in the study (12.7 h-1 ). Coincident with the formation of gas-phase species, particulate chlorine levels also rise. Cl2 , ClNO2 , NHCl2 , and NCl3 exist in the headspace of the bleach solution, whereas HOCl was only observed after floor washing. HOCl decays away 1.4 times faster than the air exchange rate, indicative of uptake onto room surfaces, and consistent with the well-known chlorinating ability of HOCl. Photochemical box modeling captures the temporal profiles of Cl2 and HOCl very well and indicates that the OH, Cl, and ClO gas-phase radical concentrations in the indoor environment could be greatly enhanced (>106 and 105  cm-3 for OH and Cl, respectively) in such washing conditions, dependent on the amount of indoor illumination.

A novel system for expansion and delivery of human keratinocytes for the treatment of severe cutaneous injuries using microcarriers and compressed collagen.

Cell therapy with autologous or allogeneic keratinocytes applied as a single-cell suspension is well established in clinical practice in the treatment of severe burn injuries to augment epithelial barrier restoration. Yet, the application of cell sprays can lead to significant cell loss owing to lack of adhesion of cell suspension to the wound bed. The development of a robust and controllable method of transplanting cells onto the wound bed is yet to be established. The ability to control adhesion and distribution of cells by using a cell carrier embedded in a biodegradable scaffold could significantly improve the treatment of cutaneous wounds with keratinocyte cell therapy. Several microcarrier-based systems for expanding keratinocytes already exist. A new method for expansion of human keratinocytes in a feeder-free, defined medium system on microcarriers has been developed. The cells retained their basal, proliferative phenotype after rapid expansion in a clinically relevant time-frame. The cell-laden microcarriers were further incorporated into collagen scaffolds fabricated by plastic compression. When cultured in vitro, cells continued to proliferate and migrate along the surface of the collagen scaffold. Using an in vitro wound bed model, cells were observed to form mostly single cell layers and in some areas multiple cell layers within 8 days, while retaining their basal, proliferative phenotype, indicating the suitability of this cell transplantation method to improve epithelial barrier restoration. This advanced cell expansion and delivery method for cutaneous cell therapy provides a flexible tool for use in clinical application. Copyright © 2017 John Wiley & Sons, Ltd.

Recent developments in delivery of nucleic acid-based antiviral agents.

Rapid advances in viral genomics, gene function and regulation, as well as in rational drug design, have led to the development of gene-based drugs that can induce protective antiviral immunity, interfere with viral replication, suppress viral gene expression or cleave viral mRNAs. Several such drug candidates have been developed in recent years against various viruses including HIV. Although gene-based agents show promise as anti-viral agents their therapeutic efficacy may be restricted by limited delivery to intracellular sites of viral replication and in vivo nuclease degradation. Enhancement of the efficacy of gene-based drugs by encapsulation within liposomes or insertion within viral vectors has been evaluated. This review will highlight recent developments in delivery systems used to target nucleic acid-based drugs into sites of viral replication, therefore avoiding potential drug toxicity in non-viral infected organs. Liposome-encapsulation and insertion of nucleic acid-based drugs within viral vectors can significantly enhance antiviral efficacies. Viral vector-mediated therapy usually results in greater expression of the gene-based drug than with liposome delivery, however significant safety concerns have been raised in regards to viral vector therapies. Research is ongoing to increase drug delivery to the desired target cells while eliminating adverse side effects.

Rapid immunofiltration assay of Newcastle disease virus using a silicon sensor.

A rapid nonradioactive sandwich immunoassay which utilizes biotin-streptavidin mediated filtration capture of immune complexes in conjunction with a silicon sensor was developed for the detection of virus. Using purified Newcastle disease virus as a model, the lower limits of detection (LOD) were determined for a number of immunoassay configurations employing both monoclonal and polyclonal antibodies. The LODs ranged from 1.3 ng/ml (sample volume of 100 microliter) for an incubation of 60 min to 400 ng/ml for a 1 min incubation. The sandwich immune complexes were formed from one-step incubation of antibody and antigen. No 'hook' effects were observed over a wide range of analyte concentrations. The assays were easy to perform and required a total time equal to the incubation period plus about 5 min. The assay format is suitable for virus, bacteria and protein antigens. New assays can be developed and optimized readily, often within 1 day.

Effect of liposome-encapsulation on immunomodulating and antiviral activities of interferon-gamma 1.

The effect of liposome-encapsulation on the immunomodulating and antiviral activities of interferon-gamma (IFN-gamma) was evaluated in this study. The immunomodulating activity was measured by increases in phagocytic activity and in nitric oxide production by peritoneal macrophages from mice treated with both free and LIP-IFN-gamma (4000 U/mouse, intraperitoneal injection). Resident peritoneal macrophages harvested from mice treated with free unencapsulated IFN-gamma or muramyl dipeptide showed significant increases in macrophage yield, and enhanced ability to phagocytize zymosan particles. In mice treated with liposome-encapsulated IFN-gamma (LIP-IFN-gamma), both macrophage yield and phagocytic activity further increased by 2-fold over unencapsulated IFN-Y. In addition, the activation of peritoneal macrophages with LIP-IFN-gamma showed enhanced production of NO when the cells were cultured ex vivo. Using a murine respiratory influenza infection model, intranasally administered LIP-IFN-gamma conferred protection to 70% in mice challenged intranasally with 10 LD50 doses of influenza A/PR/8 virus compared with a 20% survival rate using free IFN-gamma. Together these results suggest that liposome-encapsulation increases the immunomodulating and antiviral activities of IFN-gamma. Liposome-encapsulation of IFN-gamma may provide additional therapeutic advantages by reducing IFN-gamma toxicity while prolonging its body retention.

Sensitive avidin-biotin amplified fluorogenic enzyme immunoassay using biotinylated monoclonal antibodies for the identification and quantitation of virus.

A highly sensitive amplified fluorogenic enzyme-linked immunosorbent assay (FELISA), which utilizes the high affinity interaction of the vitamin biotin for the multiple binding sites on the glycoprotein avidin, was developed for the detection and identification of a model virus, Newcastle disease virus (NDV). Monoclonal antibodies (MCA) directed against the virus were purified and labelled with biotin. Biotinylated MCA was then used with avidin-labelled enzyme and a fluorogenic substrate to detect NDV adsorbed directly on nitrocellulose membranes. Reagents were standardized and, using purified virus, the theoretical lower limit of test sensitivity of the amplified FELISA was determined to be 1 fg/ml of test sample (50 ag/well). The specificity of the amplified FELISA was evaluated by challenging the assay system with homologous and heterologous strains of NDV, and with other serologically related and unrelated viruses. The test was simple to perform and multiple samples could be conveniently assayed with results obtainable in 3-4 h.

Sensitive fluorogenic enzyme immunoassay on nitrocellulose membranes for quantitation of virus.

A highly sensitive fluorogenic enzyme-linked immunosorbent assay (FELISA), which utilizes nitrocellulose membranes as solid phase support, has been developed for the detection and identification of virus in clinical samples. Reagents were standardized and, using purified Newcastle disease virus (NDV) as a model, the theoretical lower limits of test sensitivity of the FELISA were compared, in both "sandwich" and "indirect" formats, to those of a comparable chromogenic enzyme-linked immunosorbent assay (CELISA). Of the systems evaluated, the "sandwich" FELISA exhibited maximum sensitivity and detected 10 fg of purified virus protein per milliliter of test sample (500 ag per test volume). Specificity of the "sandwich" FELISA was evaluated by challenging the system with heterologous strains of NDV and with other serologically related and unrelated viruses. In a clinical trial in which fecal materials from chickens undergoing vaccination with NDV were assayed directly by FELISA, the virus was detected from the first to approximately the tenth day post-vaccination. The test is simple to perform and results can be obtained in approximately 4 h.

Repeated use of two Chlorella species, C. vulgaris and WW1 for cyclic nickel biosorption.

Two living Chlorella species were used to remove nickel from solution containing 30 micrograms Ni ml-1 in 10 successive cycles. The present study also examined the continued viability of these two algal species after repeated exposure to nickel. The two species of Chlorella were Chlorella vulgaris (commercially available) and WW1 (indigenous species isolated from domestic sewage and was tentatively identified as Chlorella miniata). The nickel removal percentage of WW1 cells was maintained at around 85% in the first five cycles, then declined slightly from the fifth cycle onwards, and finally achieved around 70% removal at the end of the 10th cycle. On the contrary, the removal efficiency of C. vulgaris declined from 50 to 30% during the 10 cycles of nickel bisorption. At the end of these 10 successive cycles, WW1 accumulated a substantial amount of Ni2+ (the cumulative cellular Ni concentration was 0.92% dry w.), while the value was only 0.17% in the case of C. vulgaris. These results suggest that the local isolate, WW1, had more consistent and satisfactory ability for removing Ni than the commercial C. vulgaris. Both algal species were still capable of dividing after each nickel treatment cycle, suggesting that the cells were not killed even when significant amounts of nickel were adsorbed/absorbed. However, Ni exposure adversely affected the physiological activity of algal cells as reflected by the decline in division rate and chlorophyll-a activity in both species. Such negative effects became more obvious as the number of cyclic treatments was increased. Nevertheless, WW1 cells appeared to recover from nickel treatment when re-cultivated in commercial medium for 2 weeks.

Revegetation of lagoon ash using the legume species Acacia auriculiformis and Leucaena leucocephala.

A greenhouse study was conducted to evaluate the potential use of two legume species, Acacia auriculiformis and Leucaena leucocephala for growth on ameliorated lagoon ash with or without nitrogen (N(2))-fixing bacteria inoculation. Even though amendments of 30% (w/w) vermiculite or with sewage sludge compost were added to improve the chemical and physical limitations of lagoon ash, significant suppressions in biomass and plant nutrient content were found with ameliorated lagoon ash in comparison to an agricultural soil. The high proportion of clay-sized (<53 microm) ash particles limited root growth. In addition, heavy metal toxicity was a possible factor contributing to poor seedling growth. Higher plant productivity resulted from the sewage sludge compost-amended lagoon ash than with vermiculite due to a greater contribution of plant nutrients in the compost. Nodulation was inhibited in ameliorated lagoon ash but not in agricultural soil. High pH and electrical conductivity and elevated toxic metals may be important parameters that limit bacterial activity. Both species showed potential to establish on amended lagoon ash, with Acacia auriculiformis being the best adapted.

Liposomes targeted to deliver antisecretory agents to jejunal mucosa.

The B subunit of cholera toxin has been covalently attached to the surface of liposomes made from a mixture of phosphatidylethanolamine, phosphatidylcholine and cholesterol. Adenylate cyclase inhibitors and chloride conductance inhibitors were encapsulated within the liposomes. These "targeted" liposomes were used to study the combined effects of this novel delivery system, and a limited number of possible antisecretory agents, on net fluid flux into the pig jejunum. A state of net secretory fluid flux was induced in isolated jejunal loops in weanling pigs by adding theophylline or cholera toxin to the lumen of the isolated loops. There was no reduction in net fluid secretion when liposome suspensions without encapsulated secretory inhibitors were added to fluid in the lumen of loops treated with theophylline. There was also no reduction in net fluid secretion when miconazole, alpha-phenylcinnamate or 5 nitro-2-(3-phenethylamino)benzoate were encapsulated within targeted liposomes added to isolated jejunal loops. The net fluid flux induced by exposure of jejunal loops to theophylline was significantly reduced by adding targeted liposomes containing 2'-deoxy-3'-AMP. The reduction involved a reversal of net secretory fluid flux to an absorptive value. The net fluid secretory response to treatment of loops with cholera toxin was also inhibited by treating loops with targeted liposomes containing 2'-deoxy-3'-AMP. However, the reversal of secretion was less complete for secretion induced by cholera toxin than for secretion induced by theophylline. The reduced antisecretory efficacy versus cholera toxin was not improved by encapsulating higher concentrations of 2'-deoxy-3'-AMP.(ABSTRACT TRUNCATED AT 250 WORDS)

Production and partial characterization of hybridoma clones secreting monoclonal antibodies against Francisella tularensis.

Hybridomas secreting monoclonal antibodies against Francisella tularensis cellular antigens were produced and characterized. These monoclonal antibodies reacted with F. tularensis in ELISA but not by immunoblot, indicating that the antibodies are directed against conformational epitopes. One of these monoclonal antibodies was directed against outer membrane protein (OMP) components, and the remainder are likely directed against capsular components. The OMP-specific monoclonal antibodies are F. tularensis-specific in contrast to the others which cross-react with a number of other bacterial species. These OMP-specific monoclonal antibodies are of the IgG1 subclass, and may prove to be a useful diagnostic tool for detection and identification of F. tularensis in clinical materials.

Epitope specificity of monoclonal antibodies against Newcastle disease virus: competitive fluorogenic enzyme immunoassay.

A test to determine the epitope specificity of monoclonal antibodies (MCA) was developed for hybridoma clones producing antibodies against Newcastle disease virus (NDV). The virus was first immobilized on nitrocellulose membranes of Millititer HA plates. Dilutions of MCA were then added, singly, or simultaneously in pairs, and bound antibody was quantitated with alkaline phosphatase-labelled detector antibody and a fluorogenic substrate, 4-methylumbelliferyl phosphate (4-MUP). Fluorescence count was measured fluorometrically. Additivity indices were calculated and plotted against dilutions of paired MCA. Antibodies that recognized identical epitopes displayed non-additivity at saturating antibody dilutions, followed by partial additivity and by total additivity at low, non-saturating dilutions. In contrast, MCA that recognized distinct epitopes exhibited total additivity throughout the curve. MCA that exhibited partial additivity were interpreted as competing for overlapping shared epitopes, or, distinct epitopes in close proximity, resulting in steric hinderance.

Pharmacokinetics study of a novel chimeric single-chain variable fragment antibody against western equine encephalitis virus.

A novel recombinant single-chain fragment variable (scFv) antibody against western equine encephalitis (WEE) virus has been previously constructed and partially characterized. The RS10B5huFc antibody was made by fusing an anti-WEE scFv to a human heavy-chain IgG1 constant region. The RS10B5huFc antibody was functional in binding to WEE virus in enzyme-linked immunosorbent assays (ELISAs), and the Fc domain of the antibody was capable of effector functions, such as binding to protein G and human complement. In this study, the RS10B5huFc antibody was further characterized by BIAcore analyses and was found to possess a binding affinity to a WEE virus epitope (K[D] = 9.14 x 10(-6) M), 4.5-fold lower than its parental mouse monoclonal antibody (MAb) 10B5 E7E2 (K[D] = 2 x 10(-6) M). No cross-reactivity was found between the RS10B5huFc antibody and three other alphaviruses (Sindbis virus [SIN], Venezuelan equine encephalitis [VEE] virus, and eastern equine encephalitis [EEE] virus). Pharmacokinetics studies showed that the RS10B5huFc antibody (free and encapsulated) was found to be retained in the lungs of mice for greater than 48 h when administered intranasally. In contrast, when administered intramuscularly to mice, the RS10B5huFc antibody was not detected in the lungs and only found in the liver and kidneys.

Characteristics of six newborn infants with postnatal findings of severe intracranial haemorrhage.

OBJECTIVE: The objective of this study was to study the characteristics of newborn infants with postnatal findings of severe neonatal intracranial haemorrhage. METHODS: All the records of babies who underwent surgery from 1997 to 2002 for intracranial haemorrhage were reviewed. These were correlated with their antenatal records to see if fetal intracranial haemorrhage had been detected at the 20 weeks' screening scan or any other incidental scan e.g. growth scan. The perinatal records were also reviewed to see if there was associated birth trauma such as instrumentation or obstetric manoeuvres at delivery. RESULTS: Six cases of severe intracranial haemorrhage were diagnosed postnatally. Of these, only 1 case was detected antenatally on ultrasound scan. None of the cases were due to birth trauma. Three babies were found to have clotting factor deficiency. One of them subsequently developed cerebral palsy. One baby was diagnosed to have alloimmune thrombocytopenia. One case underwent an emergency Caesarean section for non-reassuring fetal status. Extensive intracranial haemorrhage, attributed to hypoxia, was found. The baby died. CONCLUSIONS: Our study suggests that neonatal intracranial haemorrhages are not exclusively due to birth trauma. The study also shows that fetal intracranial haemorrhage may not be detected antenatally by the routine practice. The causes in our study included clotting deficiency, alloimmune thrombocytopenia and hypoxia.

Antibody response in goats vaccinated with liposome-adjuvanted Clostridium perfringens type D epsilon toxoid.

A trial was performed using 20 goats to evaluate the antibody responses to a liposome-adjuvanted Clostridium perfringens epsilon toxoid vaccine (LIPV). The antibody response was compared with those produced by epsilon toxoid vaccines prepared using aluminium hydroxide (ALV) and incomplete Freud's adjuvant (FAV). The animals were allocated to four groups at the beginning of the trial. The animals in group 1 were vaccinated with ALV, while the animals in group 2 received FAV and those in groups 3 and 4 were vaccinated with LIPV. The animals in groups 1 to 3 received three doses of the corresponding vaccine at intervals of three weeks, while those in group 4 received only 1 dose of vaccine at the beginning of the trial. A blood sample was obtained from all the goats at the beginning of the trial and then weekly for 8 weeks. The samples were analysed for epsilon toxoid antibodies by an indirect ELISA technique. No major clinical abnormalities were observed in the animals after vaccination, with the exception of those that received the FAV, which experienced transient lameness. The highest antibody response was observed in the animals vaccinated with FAV, but they presented moderate to severe inflammatory tissue reactions at the injection site. Moderately high antibody responses were obtained with the ALV, with which only minor local reactions were observed. No significant antibody responses were obtained with the LIPV, nor were local reactions observed.

Antiviral role of toll-like receptor-3 agonists against seasonal and avian influenza viruses.

The divergence and antigenic shifts in influenza viruses represent significant challenges for the development of effective vaccines and antiviral drugs against influenza viruses. In view of current challenges and/or deficiencies in the influenza pandemic influenza preparedness, novel antiviral strategies which are robust and can respond to constant viral mutations, are particularly needed to combat future pandemic threats. Toll-like receptor-3 (TLR-3) is an integral part of the host's innate immune system and serves as an important signaling pathway for the recognition of dsRNA for the triggering of antiviral and inflammatory responses to combat viral infections. This review examines dsRNA including Poly ICLC and liposome-encapsulated Poly ICLC (LE Poly ICLC) as TLR-3 agonists for their antiviral activity against seasonal and highly pathogenic avian influenza (HPAI) viruses. Furthermore, their roles in attenuating the antiviral and inflammatory cytokines in the host will also be explored. Preclinical studies in experimental animals suggest Poly ICLC and liposome-encapsulated Poly ICLC are safe and offer broad-spectrum protection against both seasonal and HPAI viruses, as well as other respiratory viruses including respiratory syncytial virus and SARS. Preliminary results from recent studies suggest these drugs up-regulate the production of interferons (-alpha, -beta, and -gamma), and tumor necrosis factor (TNF-alpha) but downregulate some proinflammatory cytokines including IL-2 and IL-4. Taken together, these results suggest these TLR-3 agonists have a promising role to play as safe, effective and broad-spectrum anti-influenza drugs that could complement other antiviral drugs to combat seasonal, zoonotic and pandemic influenza viruses. The clinical safety of these drugs and their efficacy in pre-clinical studies may provide sufficient justification for regulatory agencies to consider their fast track development for use in future outbreaks of pandemic influenza or of other emerging respiratory pathogens.

Activation of toll-like receptor signaling pathway for protection against influenza virus infection.

This study aims to evaluate the antiviral role of nucleic acid-based agonists for the activation of toll-like receptor (TLR) signaling pathways, and its protective role in respiratory influenza A virus infections. TLR-3 is expressed on myeloid dendritic cells, respiratory epithelium, and macrophages, and appears to play a central role in mediating both the antiviral and inflammatory responses of the innate immunity in combating viral infections. Influenza viruses can effectively inhibit the host's ability to produce interferons, and thereby suppress the immune system's antiviral defence mechanisms. Poly ICLC is a synthetic double stranded RNA comprising of polyriboinosinic-poly ribocytidylic acid (Poly IC) stabilized with l-lysine (L) and carboxymethylcellulose (C). Poly ICLC and liposome-encapsulated Poly ICLC (LE Poly ICLC) are TLR-3 agonists and are potent inducer of interferons and natural killer cells. Intranasal pre-treatment of mice with Poly ICLC and LE Poly ICLC provided high level of protection against lethal challenge with a highly lethal avian H5N1 influenza (HPAI) strain (A/H5N1/chicken/Henan clade 2), and against lethal seasonal influenza A/PR/8/34 [H1N1] and A/Aichi/2 [H3N2] virus strains. The duration of protective antiviral immunity to multiple lethal doses of influenza virus A/PR/8/34 virus had been previously found to persist for up to 3 weeks in mice for LE Poly ICLC and 2 weeks for Poly ICLC. Similarly, pre-treatment of mice with CpG oligonucleotides (TLR-9 agonist) was also found to provide complete protection against influenza A/PR/8/34 infection in mice. RT-PCR analysis of lung tissues of mice treated with Poly ICLC and LE Poly ICLC revealed upregulation of TLR-3 mRNAs gene expression. Taken together, these results do support the potential role of TLR-3 and TLR-9 agonists such as Poly ICLC and LE Poly ICLC in protection against lethal seasonal and HPAI virus infection.