Hamsters Protected from SARS-CoV-2 Delta Variant Challenge after Two Doses of Adjuvanted SARS-CoV-2 Recombinant Spike Protein (S-2P) and One Dose of Beta S-2P.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern negatively impact the effectiveness of vaccines. In this study, we challenge hamsters with the delta variant after 2- or 3-dose inoculations with SARS-CoV-2 vaccines constructed from stabilized prefusion spike proteins (S-2P) of Wuhan (W) and beta (B) variants. Compared to 3 doses of W S-2P, 2 doses of W S-2P followed by a third dose of B S-2P induced the highest neutralizing antibody titer against live SARS-CoV-2 virus and enhanced neutralization of omicron variant pseudovirus. Reduced lung live virus titer and pathology suggested that all vaccination regimens protect hamsters from SARS-CoV-2 delta variant challenge.

Evaluating the Neutralizing Ability of a CpG-Adjuvanted S-2P Subunit Vaccine Against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Variants of Concern.

BACKGROUND: Variants of concern (VoCs) have the potential to diminish the neutralizing capacity of antibodies elicited by vaccines. MVC-COV1901 is a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine consisting of recombinant prefusion stabilized spike protein S-2P adjuvanted with CpG 1018 and aluminum hydroxide. We explored the effectiveness of MVC-COV1901 against the VoCs. METHODS: Serum samples were taken from rats and phase 1 clinical trial human subjects immunized with a low, medium, or high dose of MVC-COV1901. The neutralizing titers of serum antibodies were assayed with pseudoviruses coated with the SARS-CoV-2 spike protein of the wild-type (WT), D614G, Alpha, or Beta variants. RESULTS: Rats vaccinated twice with vaccine containing high doses of antigen retained high levels of neutralization activity against the Beta variant, albeit with a slight reduction compared to WT. After the third dose, neutralizing titers against the Beta variant were noticeably enhanced regardless of the amount of antigen used for immunization. In humans, vaccinated phase 1 subjects still showed appreciable neutralization abilities against the D614G, Alpha, and Beta variants, although neutralizing titers were significantly reduced against the Beta variant. CONCLUSIONS: Two doses of MVC-COV1901 were able to elicit neutralizing antibodies against SARS-CoV-2 variants with an overall tendency of inducing higher immune response at a higher dose level. The neutralizing titers to the Beta variant in rats and humans were lower than those for WT and the Alpha variant. An additional third dose in rats was able to partially compensate for the reduction in neutralization against the Beta variant. We have demonstrated that immunization with MVC-COV1901 was effective against VoCs.

Oxaliplatin resistance in colorectal cancer cells is mediated via activation of ABCG2 to alleviate ER stress induced apoptosis.

Oxaliplatin (OXA), is a third generation platinum drug used as first-line chemotherapy in colorectal cancer (CRC). Cancer cells acquires resistance to anti-cancer drug and develops resistance. ATP-binding cassette (ABC) drug transporter ABCG2, one of multidrug resistance (MDR) protein which can effectively discharge a wide spectrum of chemotherapeutic agents out of cancer cells and subsequently reduce the intracellular concentration of these drugs. Role of ABCG2 and plausible molecular signaling pathways involved in Oxaliplatin-Resistant (OXA-R) colon cancer cells was evaluated in the present study. OXA resistant LoVo cells was developed by exposing the colon cells to OXA in a dose-dependent manner. Development of multi drug resistance in OXA-R cells was confirmed by exposing the resistance cells to oxaliplatin, 5-FU, and doxorubicin. OXA treatment resulted in G2 phase arrest in parental LoVo cells, which was overcome by OXA-R LoVo cells. mRNA and protein expression of ABCG2 and phosphorylation of NF-κB was significantly higher in OXA-R than parental cells. Levels of ER stress markers were downregulated in OXA-R than parental cells. OXA-R LoVo cells exposed to NF-κB inhibitor QNZ effectively reduced the ABCG2 and p-NF-κB expression and increased ER stress marker expression. On other hand, invasion and migratory effect of OXA-R cells were found to be decreased, when compared to parental cells. Metastasis marker proteins also downregulated in OXA-R cells. ABCG2 inhibitor verapamil, downregulate ABCG2, induce ER stress markers and induces apoptosis. In vivo studies in nude mice also confirms the same.

Lupeol alters ER stress-signaling pathway by downregulating ABCG2 expression to induce Oxaliplatin-resistant LoVo colorectal cancer cell apoptosis.

Colorectal cancer (CRC) is one of the most common cancers and causes of cancer-related death. There are several first-line chemotherapeutic drugs used to treat CRC. Oxaliplatin (OXA) is an alkylating cytotoxic agent that is usually combined with other chemotherapeutic drugs to treat stage II and stage III CRC. However, cancer cells commonly acquire multidrug resistance (MDR), which is a major obstruction to cancer treatment. Recent studies have shown that natural components from traditional Chinese medicine or foods that have many biological functions may be new adjuvant therapies in clinical trials. We challenged LoVo CRC cell lines with OXA in a dose-dependent manner to create an OXA-resistant model. The expression of ABCG2 was significantly higher, and levels of endoplasmic reticulum (ER) stress markers were lower than those Parental cells. However, Lupeol, which is found in fruits and vegetables, has been shown to have bioactive properties, including anti-tumor properties that are relevant to many diseases. In our study, Lupeol downregulated cell viability and activated cell apoptosis. Moreover, Lupeol decreased the expression of ABCG2 and activated ER stress to induce OXA-resistant cell death. Importantly, the anti-tumor effect of Lupeol in OXA-resistant cells was higher than that of LoVo Parental cells. In addition, we also confirmed our results with a xenograft animal model, and the tumor size significantly decreased after Lupeol injections. Our findings show that Lupeol served as a strong chemoresistant sensitizer and could be a new adjuvant therapy method for chemoresistant patients.

Hepatitis B virus nucleocapsid but not free core antigen controls viral clearance in mice.

We have recently shown that hepatitis B virus (HBV) core antigen (HBcAg) is the major viral factor for HBV clearance using a hydrodynamics-based mouse model. Knockout of HBcAg hampers the development of antiviral immune responses and thus promotes HBV persistence. Here, we further demonstrated that only in the capsid form, but not the free or dimer form, can HBcAg exert its contributory role in HBV clearance. HBcAg is the main structural protein of HBV icosahedral nucleocapsid. A mutant HBV DNA which expresses an assembly-defective HBcAg, HBcAgY132A, surprisingly prolonged HBV surface antigenemia in both C57BL/6 and BALB/c mice without affecting viral transcription and translation. This result was not due to a loss of the possible immune epitope caused by the single-amino-acid substitution of HBcAg. Moreover, the particular HBV mutant failed to induce robust humoral and cellular immunity against HBV. These data revealed the requirement of capsid structure for inducing adequate immunity that leads to HBV clearance in mice.

Hepatitis B virus core antigen determines viral persistence in a C57BL/6 mouse model.

We recently developed a mouse model of hepatitis B virus (HBV) persistence, in which a single i.v. hydrodynamic injection of HBV DNA to C57BL/6 mice allows HBV replication and induces a partial immune response, so that about 20-30% of the mice carry HBV for more than 6 months. The model was used to identify the viral antigen crucial for HBV persistence. We knocked out individual HBV genes by introducing a premature termination codon to the HBV core, HBeAg, HBx, and polymerase ORFs. The specific-gene-deficient HBV mutants were hydrodynamically injected into mice and the HBV profiles of the mice were monitored. About 90% of the mice that received the HBcAg-mutated HBV plasmid exhibited high levels of hepatitis B surface antigenemia and maintained HBsAg expression for more than 6 months after injection. To map the region of HBcAg essential for viral clearance, we constructed a set of serial HBcAg deletion mutants for hydrodynamic injection. We localized the essential region of HBcAg to the carboxyl terminus, specifically to the 10 terminal amino acids (HBcAg176-185). The majority of mice receiving this HBV mutant DNA did not elicit a proper HBcAg-specific IFN-gamma response and expressed HBV virions for 6 months. These results indicate that the immune response triggered in mice by HBcAg during exposure to HBV is important in determining HBV persistence.

Glucocorticoid Receptor Maintains Vasopressin Responses in Kidney Collecting Duct Cells.

Water permeability of the kidney collecting ducts is regulated in part by the amount of the molecular water channel protein aquaporin-2 (AQP2), whose expression, in turn, is regulated by the pituitary peptide hormone vasopressin. We previously showed that stable glucocorticoid receptor knockdown diminished the vasopressin-induced Aqp2 gene expression in the collecting duct cell model mpkCCD. Here, we investigated the pathways regulated by the glucocorticoid receptor by comparing transcriptomes of the mpkCCD cells with or without stable glucocorticoid receptor knockdown. Glucocorticoid receptor knockdown downregulated 5,394 transcripts associated with 55 KEGG pathways including "vasopressin-regulated water reabsorption," indicative of positive regulatory roles of these pathways in the vasopressin-induced Aqp2 gene expression. Quantitative RT-PCR confirmed the downregulation of the vasopressin V2 receptor transcript upon glucocorticoid receptor knockdown. Glucocorticoid receptor knockdown upregulated 3,785 transcripts associated with 42 KEGG pathways including the "TNF signaling pathway" and "TGFß signaling pathway," suggesting the negative regulatory roles of these pathways in the vasopressin-induced Aqp2 gene expression. Quantitative RT-PCR confirmed the upregulation of TNF and TGFß receptor transcripts upon glucocorticoid receptor knockdown. TNF or TGFß inhibitor alone, in the absence of vasopressin, did not induce Aqp2 gene transcription. However, TNF or TGFß blunted the vasopressin-induced Aqp2 gene expression. In particular, TGFß reduced vasopressin-induced increases in Akt phosphorylation without inducing epithelial-to-mesenchymal transition or interfering with vasopressin-induced apical AQP2 trafficking. In summary, our RNA-seq transcriptomic comparison revealed positive and negative regulatory pathways maintained by the glucocorticoid receptor for the vasopressin-induced Aqp2 gene expression.

Protection of hamsters challenged with SARS-CoV-2 after two doses of MVC-COV1901 vaccine followed by a single intranasal booster with nanoemulsion adjuvanted S-2P vaccine.

Intramuscular vaccines have greatly reduced hospitalization and death due to severe COVID-19. However, most countries are experiencing a resurgence of infection driven predominantly by the Delta and Omicron variants of SARS-CoV-2. In response, booster dosing of COVID-19 vaccines has been implemented in many countries to address waning immunity and reduced protection against the variants. However, intramuscular boosting fails to elicit mucosal immunity and therefore does not solve the problem of persistent viral carriage and transmission, even in patients protected from severe disease. In this study, two doses of stabilized prefusion SARS-CoV-2 spike (S-2P)-based intramuscular vaccine adjuvanted with Alum/CpG1018, MVC-COV1901, were used as a primary vaccination series, followed by an intranasal booster vaccination with nanoemulsion (NE01)-adjuvanted S-2P vaccine in a hamster model to demonstrate immunogenicity and protection from viral challenge. Here we report that this vaccination regimen resulted not only in the induction of robust immunity and protection against weight loss and lung pathology following challenge with SARS-CoV-2, but also led to increased viral clearance from both upper and lower respiratory tracts. Our findings showed that intramuscular MVC-COV1901 vaccine followed by a booster with intranasal NE01-adjuvanted vaccine promotes protective immunity against both viral infection and disease, suggesting that this immunization protocol may offer a solution in addressing a significant, unmet medical need for both the COVID-19 and future pandemics.

Intranasal nanoemulsion adjuvanted S-2P vaccine demonstrates protection in hamsters and induces systemic, cell-mediated and mucosal immunity in mice.

With the rapid progress made in the development of vaccines to fight the SARS-CoV-2 pandemic, almost >90% of vaccine candidates under development and a 100% of the licensed vaccines are delivered intramuscularly (IM). While these vaccines are highly efficacious against COVID-19 disease, their efficacy against SARS-CoV-2 infection of upper respiratory tract and transmission is at best temporary. Development of safe and efficacious vaccines that are able to induce robust mucosal and systemic immune responses are needed to control new variants. In this study, we have used our nanoemulsion adjuvant (NE01) to intranasally (IN) deliver stabilized spike protein (S-2P) to induce immunogenicity in mouse and hamster models. Data presented demonstrate the induction of robust immunity in mice resulting in 100% seroconversion and protection against SARS-CoV-2 in a hamster challenge model. There was a significant induction of mucosal immune responses as demonstrated by IgA- and IgG-producing memory B cells in the lungs of animals that received intranasal immunizations compared to an alum adjuvanted intramuscular vaccine. The efficacy of the S-2P/NE01 vaccine was also demonstrated in an intranasal hamster challenge model with SARS-CoV-2 and conferred significant protection against weight loss, lung pathology, and viral clearance from both upper and lower respiratory tract. Our findings demonstrate that intranasal NE01-adjuvanted vaccine promotes protective immunity against SARS-CoV-2 infection and disease through activation of three arms of immune system: humoral, cellular, and mucosal, suggesting that an intranasal SARS-CoV-2 vaccine may play a role in addressing a unique public health problem and unmet medical need.

Safety and immunogenicity of a Recombinant Stabilized Prefusion SARS-CoV-2 Spike Protein Vaccine (MVC-COV1901) Adjuvanted with CpG 1018 and Aluminum Hydroxide in healthy adults: A Phase 1, dose-escalation study.

BACKGROUND: This was a phase 1, dose-escalation open-label trial to evaluate the safety and immunogenicity of MVC-COV1901, a SARS-CoV-2 S-2P protein vaccine adjuvanted with aluminum hydroxide and CpG 1018. METHODS: Between September 28 and November 13 2020, 77 participants were screened. Of these, 45 healthy adults from 20 to 49 years of age were to be administered two doses of MVC-COV1901 in doses of 5 µg, 15 µg, or 25 µg of spike protein at 28 days apart. There were 15 participants in each dose group; all were followed for 28 days after the second dose at the time of the interim analysis. Adverse events and laboratory data were recorded for the safety evaluation. Blood samples were collected for humoral, and cellular immune response at various time points. Trial Registration: ClinicalTrials.gov NCT04487210. FINDINGS: Solicited adverse events were mostly mild and similar. No subject experienced fever. After the second dose, the geometric mean titers (GMTs) for SARS-CoV-2 spike-specific immunoglobulin G were 7178.2, 7746.1, 11,220.6 in the 5 µg, 15 µg, and 25 µg dose groups, respectively. The neutralizing activity were detected in both methods. (Day 43 GMTs, 538.5, 993.1, and 1905.8 for pseudovirus; and 33.3, 76.3, and 167.4 for wild-type virus). The cellular immune response induced by MVC-COV1901 demonstrated substantially higher numbers of IFN-γ- producing cells, suggesting a Th1-skewed immune response. INTERPRETATION: The MVC-COV1901 vaccine was well tolerated and elicited robust immune responses and is suitable for further development. FUNDING: Medigen Vaccine Biologics Corporation.

CpG-adjuvanted stable prefusion SARS-CoV-2 spike protein protected hamsters from SARS-CoV-2 challenge.

The COVID-19 pandemic presents an unprecedented challenge to global public health. Rapid development and deployment of safe and effective vaccines are imperative to control the pandemic. In the current study, we applied our adjuvanted stable prefusion SARS-CoV-2 spike (S-2P)-based vaccine, MVC-COV1901, to hamster models to demonstrate immunogenicity and protection from virus challenge. Golden Syrian hamsters immunized intramuscularly with two injections of 1 µg or 5 µg of S-2P adjuvanted with CpG 1018 and aluminum hydroxide (alum) were challenged intranasally with SARS-CoV-2. Prior to virus challenge, the vaccine induced high levels of neutralizing antibodies with 10,000-fold higher IgG level and an average of 50-fold higher pseudovirus neutralizing titers in either dose groups than vehicle or adjuvant control groups. Six days after infection, vaccinated hamsters did not display any weight loss associated with infection and had significantly reduced lung pathology and most importantly, lung viral load levels were reduced to lower than detection limit compared to unvaccinated animals. Vaccination with either 1 µg or 5 µg of adjuvanted S-2P produced comparable immunogenicity and protection from infection. This study builds upon our previous results to support the clinical development of MVC-COV1901 as a safe, highly immunogenic, and protective COVID-19 vaccine.

Development of CpG-adjuvanted stable prefusion SARS-CoV-2 spike antigen as a subunit vaccine against COVID-19.

The COVID-19 pandemic is a worldwide health emergency which calls for an unprecedented race for vaccines and treatment. In developing a COVID-19 vaccine, we applied technology previously used for MERS-CoV to produce a prefusion-stabilized SARS-CoV-2 spike protein, S-2P. To enhance immunogenicity and mitigate the potential vaccine-induced immunopathology, CpG 1018, a Th1-biasing synthetic toll-like receptor 9 (TLR9) agonist was selected as an adjuvant candidate. S-2P in combination with CpG 1018 and aluminum hydroxide (alum) was found to be the most potent immunogen and induced high titer of neutralizing antibodies in sera of immunized mice against pseudotyped lentivirus reporter or live wild-type SARS-CoV-2. In addition, the antibodies elicited were able to cross-neutralize pseudovirus containing the spike protein of the D614G variant, indicating the potential for broad spectrum protection. A marked Th1 dominant response was noted from cytokines secreted by splenocytes of mice immunized with CpG 1018 and alum. No vaccine-related serious adverse effects were found in the dose-ranging study in rats administered single- or two-dose regimens of S-2P combined with CpG 1018 alone or CpG 1018 with alum. These data support continued development of CHO-derived S-2P formulated with CpG 1018 and alum as a candidate vaccine to prevent COVID-19 disease.

Pediatric reference intervals for several biochemical analytes in school children in Central Taiwan.

BACKGROUND/PURPOSE: Reference intervals of biochemical tests for screening for diabetes mellitus and liver and renal function among school children in Central Taiwan have never been documented. Therefore, this study aimed to establish the reference intervals for the above mentioned biochemical tests for pediatric populations. METHODS: A total of 4326 subjects, including 2029 kindergarten children, 1624 elementary-school children, 325 junior-high-school children, and 348 teachers were selected randomly in Central Taiwan. All serum alanine aminotransferase (ALT), blood urea nitrogen (BUN), creatinine (Cr) and glucose levels were determined using a Beckman Synchron CX5 analyzer. The reference intervals reflected estimates of the 2.5th-97.5th percentiles of non-parametric distributions. RESULTS: Adults had significantly higher biochemical analyte values [except for BUN/creatinine (B/C) ratio] than children had. Multiple logistic regression analysis showed that biochemical analyte values were significantly higher in male than in female subjects. The concentrations of glucose and Cr increased with age. On the contrary, the B/C ratio decreased with age. CONCLUSION: Our study provides new pediatric reference intervals (2.5th-97.5th percentiles) of 60-99 mg/dL for serum glucose concentrations, 8-38 IU/L for ALT, 0.4-1.1 mg/L for Cr, 8.7-18.0 mg/L for BUN, and 10-34 for B/C ratio. The B/C ratio in children was higher than those of adults, possibly due to that children had a higher intake of protein.

A further investigation of the good-enough level model across outcome domains and termination status.

Accurate estimations of progress in psychotherapy are necessary for therapists to identify clients at risk of deterioration and potentially reduce premature terminations. This need has resulted in a large body of literature examining the rate and trajectory of change in psychotherapy; however, few studies have tested these dose-response relationships outside of global measures of mental health. Moreover, there is a paucity of research examining the relationship between progress in treatment, treatment length, and premature termination. In this study, we conducted multivariate multilevel analyses to test the good-enough level model across the three domains of the phase model of psychotherapy: psychological symptoms, life functioning, and well-being. In addition, we tested changes in well-being, psychological symptoms, and life functioning, treatment length, and an interaction between treatment progress and treatment length as predictors of premature termination. Data for this study consisted of 438 clients who were treated by 57 therapists within a brief therapy model. Results failed to support the good-enough level model for changes in well-being, psychological symptoms, and life functioning, such that the rate of change across all three scales did not significantly vary as a function of treatment length. However, exploratory analyses revealed a significant interaction effect between changes in well-being, treatment length, and premature termination, indicating that clients who experienced high rates of change in well-being early in treatment were more likely to prematurely terminate treatment. Clinical implications and future research are discussed. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

Removal of EDTA from low pH printed-circuit board wastewater in a fluidized zero valent iron reactor.

Fluidized zero valent iron (ZVI) process was adopted to reduce ethylenediaminetetraacetic acid (EDTA) from low pH printed-circuit board (PCB) wastewater for two reasons: (1) low pH of the wastewater favoring the ZVI reaction; (2) higher ZVI utilization for fluidized process due to abrasive motion of the ZVI. The results showed that the degradation of EDTA was greatly enhanced under acidic pH, longer hydraulic detention time (HRT) and presence of dissolved oxygen (DO). Without addition of oxygen, 65% of EDTA was removed with capacity of 7.33 mg EDTA/g ZVI at pH 2, ZVI dosage of 424 g/L and HRT 10 min. With 6.8 mg/L of DO, 83% of EDTA was reduced with capacity of 19.01 mg EDTA/g ZVI for the same experimental condition. The presence of oxygen/ZVI initiated a Fenton type reaction to reduce EDTA. The end product after EDTA degradation was analyzed by high performance liquid chromoagraphy (HPLC), where propionic acid (C(2)H(5)COOH) was observed, indicating EDTA (oxidation number for carbon is 2) was oxidized to propionic acid (oxidation number for carbon is 3). Nitrogen species was also measured and the nitrogen in EDTA was converted to ammonium instead of nitrate and nitrite.

Vocational rehabilitation case manager factors associated with vocational rehabilitation service program outcomes for people with disabilities in Taiwan - an exploratory study.

Purpose The aim of this study is to explore whether/which vocational rehabilitation case manager (VRCMer) factors were significantly associated with the vocational rehabilitation service (VRS) program outcomes in Taiwan. Method This study used the 2011 VRS Program for People with Disabilities Database in a metropolitan city in Taiwan (N = 466) to do a secondary data analysis using hierarchical logistic regression. Results This study found that the employment rate and stable employment rate created by the 2011 VRS program in a metropolitan city in Taiwan were 48.7% and 42.1%, respectively. For the predictors of employment/stable employment, "occurrences of the services provided by the VRCMer" variable was definitely dominant. In addition, "level of the disability" was the second-ranking predictor, and was significantly negatively correlated with both employment and stable employment outcomes. Conclusions Vocational rehabilitation case manager factors in this study were significantly correlated with VRS program outcomes for people with disabilities in Taiwan after controlling for the clients' socio-demographic variables. The results indicate that greater input by VRCMers for people with disabilities equates to better employment outcomes in metropolitan Taiwan. Implications for Rehabilitation This is the first study to build an inferential statistical model in attempt to explain and predict the association between vocational rehabilitation case manager factors and vocational rehabilitation service program outcomes for people with disabilities in Taiwan. In cases of severe disability, a vocational rehabilitation case manager should seek out more in-kind and in-cash resources, and choose a suitable job coach to cooperate in assisting the client to become employed. Based on the findings, government has to continue implementing opportunities for people with disabilities to attain higher and better quality educational levels, for increasing their employment rate. Vocational rehabilitation case managers should raise the referral rate and cooperation with job coaches as this directly affects the quality of services and clients' employment rate.

Outcomes of home-based employment service programs for people with disabilities and their related factors--a preliminary study in Taiwan.

PURPOSE: The aim of this exploratory study is to gain an understanding of the outcomes of home-based employment service programs for people with disabilities and their related factors in Taiwan. METHOD: This study used survey method to collect 132 questionnaires. Descriptive and two-variable statistics including chi-square (χ(2)), independent sample t-test and analysis of variance were employed. RESULTS: The results found that 36.5% of the subjects improved their employment status and 75.8% of them improved in employability. Educational level and and vocational categories including "web page production", "e-commerce", "internet marketing", "on-line store" and "website set-up and management" were significantly "positively" associated with either of the two outcome indicators - change of employment status and employability. CONCLUSIONS: This study is the first evidence-based study about the outcomes of home-based employment service programs and their related factors for people with disabilities in Taiwan. The outcomes of the home-based employment service programs for people with disabilities were presented. Implications for Rehabilitation Home-based rehabilitation for people with disabilities can be effective. A programme of this kind supports participants in improving or gaining employment status as well as developing employability skills. Further consideration should be given to developing cost-effective home-based programmes and evaluating their effectiveness.

Role of TRPV1 and P2X receptors in the activation of lung vagal C-fiber afferents by inhaled cigarette smoke in rats.

Inhaled cigarette smoke (CS) triggers airway reflexes that are thought to result from the activation of lung vagal C-fiber afferents (LVCAs) via the action of reactive oxygen species in rats. We investigated the role of transient receptor potential vanilloid 1 (TRPV1) and P2X receptors in LVCA activation. Activities of LVCAs were recorded in anesthetized and artificially ventilated rats. Airway challenge of CS produced a concentration-dependent fiber stimulation. Pretreatment with dimethylthiourea [DMTU; a scavenger of hydroxyl radical (OH)], capsazepine (CPZ; a TRPV1 receptor antagonist) and iso-pyridoxalphosphate-6-azophenyl-2',5'-disulphonate (iso-PPADS; a P2X receptor antagonist) separately reduced the fiber responses by 64, 40 and 44%, respectively, whereas pretreatment with hexamethonium (a nicotinic acetylcholine receptor antagonist) failed to alter the response. A combination of CPZ and iso-PPADS exerted a greater inhibitory effect compared with the effect of either single pretreatment. However, a combination of DMTU, CPZ and iso-PPADS did not further reduce the fiber response compared with the combined effect of CPZ and iso-PPADS. It was concluded that both TRPV1 and P2X receptors, but not nicotinic acetylcholine receptors, participate in the stimulation of LVCAs by inhaled CS, possibly through the action of OH.

Cost-benefit analysis for sheltered employment service programs for people with disabilities in Taiwan - a preliminary study.

PURPOSE: The aim of this study is to do a cost-benefit analysis with monetary and non-monetary benefits for sheltered employment service programs and try to provide more evidence-based information for policy makers and practitioners to understand the outcomes of sheltered employment services. METHOD: This study analyzed 3 sheltered employment service programs for people with disabilities (2006-2007) implemented by Sunshine Social Welfare Foundation in Taiwan using cost-benefit analysis (including non-monetary benefits). Three groups were analyzed, including participants in the programs, taxpayers, and society (participants and taxpayers). RESULTS: This study found that the net social monetary benefit was $NT29,432.07 per participant per year and the benefit cost ratio was 1.43. (In 2006-2007, $US1 = $NT32.5 averagely around.) The net monetary benefit for the participants was between $NT7,890.86 and $NT91,890.86 per participant per year. On the non-monetary benefit side, the physical health (raised 7.49%), social relationship (raised 3.36%) domains, and general quality of life (raised 2.53%) improved. However, the psychological (decreased 1.51%) and working/environment (decreased 3.85%) domains backslided. In addition, the differences between pre-test and post-test average scores of all domains were not statistically significant. CONCLUSIONS: This study is the first to use monetary and non-monetary cost-benefit analysis methods to analyze sheltered employment service programs for people with disabilities in Taiwan. The findings indicated that sheltered employment service programs for people with disabilities could be efficient and beneficial for the whole society and sheltered employees/clients, and also helpful for raising their quality of lives.

PD-1 blockage reverses immune dysfunction and hepatitis B viral persistence in a mouse animal model.

Persistent hepatitis B viral (HBV) infection results in chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). Recent studies in animal models of viral infection indicate that the interaction between the inhibitory receptor, programmed death (PD)-1, on lymphocytes and its ligand (PD-L1) play a critical role in T-cell exhaustion by inducing T-cell inactivation. High PD-1 expression levels by peripheral T-lymphocytes and the possibility of improving T-cell function by blocking PD-1-mediated signaling confirm the importance of this inhibitory pathway in inducing T-cell exhaustion. We studied T-cell exhaustion and the effects of PD-1 and PD-L1 blockade on intrahepatic infiltrating T-cells in our recently developed mouse model of HBV persistence. In this mouse animal model, we demonstrated that there were increased intrahepatic PD-1-expressing CD8+ and CD4+ T cells in mice with HBV persistence, but PD-1 upregulation was resolved in mice which had cleared HBV. The Intrahepatic CD8+ T-cells expressed higher levels of PD-1 and lower levels of CD127 in mice with HBV persistence. Blockade of PD-1/PD-L1 interactions increased HBcAg-specific interferon (IFN)-γ production in intrahepatic T lymphocytes. Furthermore, blocking the interaction of PD-1 with PD-L1 by an anti-PD-1 monoclonal antibody (mAb) reversed the exhausted phenotype in intrahepatic T lymphocytes and viral persistence to clearance of HBV in vivo. Our results indicated that PD-1 blockage reverses immune dysfunction and viral persistence of HBV infection in a mouse animal model, suggesting that the anti-PD-1 mAb might be a good therapeutic candidate for chronic HBV infection.