ABSTRACT
Tumor metastasis and cancer recurrence are often a result of cell heterogeneity, where specific subpopulations of tumor cells may be resistant to radio- or chemotherapy. To investigate this physiological and phenotypic diversity, single-cell metabolomics provides a powerful approach at the chemical level, where distinct lipid profiles can be found in different tumor cells. Here, we established a highly sensitive platform using nanoflow liquid chromatography (nLC) combined with multinozzle emitter electrospray ionization mass spectrometry for more in-depth metabolomics profiling. Our platform identified 15 and 17 lipids from individual osteosarcoma (U2OS) and glioblastoma (GBM) cells when analyzing single-cell samples. Additionally, we used the functional single-cell selection (fSCS) pipeline to analyze the subpopulations of cells with a DNA damage response (DDR) in U2OS cells and fast migration in GBM cells. Specifically, we observed a down-regulation of polyunsaturated fatty acids (PUFAs) in U2OS cells undergoing DDR, such as fatty acids FA 20:3; O2 and FA 17:4; O3. Furthermore, ceramides (Cer 38:0; O3) and triglycerides (TG 36:0) were found to be down-regulated in fast-migrating GBM cells compared to the slow-migrating subpopulation. These findings suggest the potential roles of these metabolites and/or lipids in the cellular behavior of the subpopulations.
Subject(s)
Glioblastoma , Spectrometry, Mass, Electrospray Ionization , Humans , Chromatography, Liquid/methods , Spectrometry, Mass, Electrospray Ionization/methods , Metabolomics/methods , Fatty Acids, Unsaturated/metabolism , TriglyceridesABSTRACT
INTRODUCTION: While retail sales and retailer inspection studies generally indicate high compliance with state sales restrictions on Nicotine Vaping Products (NVPs) within the restricted area, studies using survey data generally indicate that most users could readily continue gaining access to restricted NVPs. Our study bridges a gap in the current literature and investigates the potential role of cross-state border purchases to evade state emergency NVP sales restrictions in 2019. AIMS AND METHODS: The study sample was restricted to NVP sales from the states neighboring Massachusetts, Rhode Island, and Washington, three states that implemented all NVP or flavored NVP sales restrictions in 2019. Among these neighboring states, the 2019 weekly county-level NVP sales by flavors (tobacco, mint/menthol, and other flavors) were compiled using Nielsen Scanner data. A quasi-experimental, comparison group pre-post study design was used to study the impacts of NVP sales restrictions on cross-state border NVP purchases. RESULTS: Weekly NVP sales for border counties significantly increased in response to the MA, RI, and WA bans for tobacco flavored (56%, 45%, 14%, respectively), menthol/mint flavored (51%, 2%, 41%, respectively), and other flavored (79%, 3%, 4%, respectively) products, compared to sales for non-border counties (all p-values < .01). CONCLUSIONS: Our study identified significant cross-state border NVP purchases in all studied states to circumvent NVP emergency sales restrictions in response to the EVALI outbreak. Policymakers should factor in these purchasing behaviors to evade sales restrictions when evaluating any future potential policies at the state or local levels. IMPLICATIONS: While retail sales data and retailer inspections indicate high compliance with Nicotine Vaping Product (NVP) flavor sales restrictions from major retail outlets, survey data obtain mixed findings on the effects of sales restrictions on vaping behaviors. Our study identified a significant increase in cross-state border NVP purchases to circumvent NVP sales restrictions in 2019, consistent across all three settings of Massachusetts, Rhode Island, and Washington. Policymakers should factor in these cross-state border NVP purchases in response to sales restrictions when evaluating any future potential NVP sales restrictions.
Subject(s)
Commerce , Flavoring Agents , Vaping , Vaping/economics , Humans , Commerce/legislation & jurisprudence , Commerce/statistics & numerical data , Massachusetts , Electronic Nicotine Delivery Systems/economics , Electronic Nicotine Delivery Systems/statistics & numerical data , Washington , Rhode Island , Nicotine , Tobacco Products/economics , Tobacco Products/legislation & jurisprudence , Consumer Behavior/statistics & numerical data , Consumer Behavior/economicsABSTRACT
AIM: To evaluate the effectiveness of magic-themed interventions in improving daily bimanual task performance in children with unilateral spastic cerebral palsy (CP) to and elucidate the variability in outcomes. METHOD: This systematic literature review searched databases including Embase, MEDLINE, Scopus, Cochrane Central, and CINAHL. Outcome measures selected for the meta-analysis included the Children's Hand-use Experience Questionnaire, its three subscales, and the Besta subscale C. The overall efficacy of magic-themed interventions was analysed using Hedges' g as the summary measure for these outcomes. Subgroup analysis compared the efficacy of different modes of training, and a meta-regression investigated the impact of training duration. RESULTS: Analyses of four studies involving 78 children showed magic-themed training significantly improved bimanual task performance (Hedges' g = 0.327, 95% confidence interval [CI] = 0.107-0.547, p = 0.004), especially in group settings (Hedges' g = 0.435, 95% CI = 0.176-0.693, p = 0.001), compared with non-significant gains from video interventions (Hedges' g = 0.041, 95% CI = -0.380 to 0.462, p = 0.850). Additionally, training duration positively correlated with performance gains (coefficient = 0.0076 per hour, p = 0.001). INTERPRETATION: Magic-themed training, especially through group sessions and extended durations, enhances bimanual skills in children with unilateral spastic CP.
ABSTRACT
During rheumatoid arthritis (RA) treatment, long-term injection of antitumor necrosis factor α antibodies (anti-TNFα Abs) may induce on-target toxicities, including severe infections (tuberculosis [TB] or septic arthritis) and malignancy. Here, we used an immunoglobulin G1 (IgG1) hinge as an Ab lock to cover the TNFα-binding site of Infliximab by linking it with matrix metalloproteinase (MMP) -2/9 substrate to generate pro-Infliximab that can be specifically activated in the RA region to enhance the selectivity and safety of treatment. The Ab lock significantly inhibits the TNFα binding and reduces the anti-idiotypic (anti-Id) Ab binding to pro-Infliximab by 395-fold, 108-fold compared with Infliximab, respectively, and MMP-2/9 can completely restore the TNFα neutralizing ability of pro-Infliximab to block TNFα downstream signaling. Pro-Infliximab was only selectively activated in the disease site (mouse paws) and presented similar pharmacokinetics (PKs) and bio-distribution to Infliximab. Furthermore, pro-Infliximab not only provided equivalent therapeutic efficacy to Infliximab but also maintained mouse immunity against Listeria infection in the RA mouse model, leading to a significantly higher survival rate (71%) than that of the Infliximab treatment group (0%). The high-selectivity pro-Infliximab maintains host immunity and keeps the original therapeutic efficiency, providing a novel strategy for RA therapy.
Subject(s)
Arthritis, Rheumatoid/drug therapy , Drug Delivery Systems/methods , Infliximab/pharmacology , Animals , Arthritis, Rheumatoid/physiopathology , Humans , Immunoglobulin G/immunology , Immunoglobulin G/therapeutic use , Infliximab/metabolism , Mice , Mice, Inbred DBA , Mice, Knockout , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Tumor Necrosis Factor-alpha/metabolismABSTRACT
The bidirectional interaction between carcinogens and gut microbiota that contributes to colorectal cancer is complicated. Reactivation of carcinogen metabolites by microbial ß-glucuronidase (ßG) in the gut potentially plays an important role in colorectal carcinogenesis. We assessed the chemoprotective effects and associated changes in gut microbiota induced by pre-administration of bacterial-specific ßG inhibitor TCH-3511 in carcinogen azoxymethane (AOM)-treated APCMin/+ mice. AOM induced intestinal ßG activity, which was reflected in increases in the incidence, formation, and number of tumors in the intestine. Notably, inhibition of gut microbial ßG by TCH-3511 significantly reduced AOM-induced intestinal ßG activity, decreased the number of polyps in both the small and large intestine to a frequency that was similar in mice without AOM exposure. AOM also led to lower diversity and altered composition in the gut microbiota with a significant increase in mucin-degrading Akkermansia genus. Conversely, mice treated with TCH-3511 and AOM exhibited a more similar gut microbiota structure as mice without AOM administration. Importantly, TCH-3511 treatment significant decreased Akkermansia genus and produced a concomitant increase in short-chain fatty acid butyrate-producing gut commensal microbes Lachnoospiraceae NK4A136 group genus in AOM-treated mice. Taken together, our results reveal a key role of gut microbial ßG in promoting AOM-induced gut microbial dysbiosis and intestinal tumorigenesis, indicating the chemoprotective benefit of gut microbial ßG inhibition against carcinogens via maintaining the gut microbiota balance and preventing cancer-associated gut microbial dysbiosis. Thus, the bacterial-specific ßG inhibitor TCH-3511 is a potential chemoprevention agent for colorectal cancer.
Subject(s)
Colorectal Neoplasms , Gastrointestinal Microbiome , Animals , Azoxymethane/toxicity , Bacteria , Carcinogenesis , Carcinogens/toxicity , Cell Transformation, Neoplastic , Colorectal Neoplasms/chemically induced , Colorectal Neoplasms/metabolism , Colorectal Neoplasms/prevention & control , Dysbiosis/prevention & control , Glucuronidase , MiceABSTRACT
Although varenicline has had a significant effect on smoking cessation in randomized clinical trials, the dose-effect of varenicline treatment for smoking cessation in real-world settings remains unclear. This study aimed to evaluate the association between the duration of varenicline prescription and smoking cessation in Taiwan after adjusting for potential confounding effects and endogeneity bias. A total of 5106 Taiwanese participants received varenicline monotherapy for smoking cessation between March 2012 and September 2016. Multinomial logistic regression (MLR) was used to analyze the association between varenicline prescription duration and smoking cessation, stratified by the frequency of smoking clinic visits and propensity scores of early stopping of smoking cessation treatment. Compared to the reference of nonquitting, longer durations of varenicline prescription were associated with the greater likelihood of immediate and complete quitting (OR = 1.08, 95% CI = 1.02-1.14) and late quitting (OR = 1.14, 95% CI = 1.07-1.20). Among those who were more likely to continue visiting smoking clinics, longer use of varenicline was significantly associated with an increase in immediate-and-complete quitting (OR = 1.19, 95% CI = 1.15-1.23) and late quitting (OR = 1.24, 95% CI = 1.20-1.28). Varenicline prescription duration was not associated with smoking cessation among smokers who visited smoking clinics once. The relationship between varenicline prescription duration and smoking cessation was modified by the frequency of smoking clinic visits and was dependent on quitting process patterns. Encouraging smokers to continue visiting the smoking cessation clinic and use medication will help smoking cessation efforts in Taiwan.
Subject(s)
Smoking Cessation , Humans , Prescriptions , Taiwan , Tobacco Use Cessation Devices , Varenicline/therapeutic useABSTRACT
RUVBL1 and RUVBL2 are highly conserved AAA ATPases (ATPases Associated with various cellular Activities) and highly relevant to the progression of cancer, which makes them attractive targets for novel therapeutic anticancer drugs. In this work, docking-based virtual screening was performed to identify compounds with activity against the RUVBL1/2 complex. Seven compounds showed inhibitory activity against the complex in both enzymatic and cellular assays. A series of pyrazolo[1,5-a]pyrimidine-3-carboxamide analogs were synthesized based on the scaffold of compound 15 with inhibitory activity and good potential for structural manipulation. Analysis of the structure-activity relationship identified the benzyl group on R2 and aromatic ring-substituted piperazinyl on R4 as essential for inhibitory activity against the RUVBL1/2 complex. Of these, compound 18, which has IC50 values of 6.0 ± 0.6 µM and 7.7 ± 0.9 µM against RUVBL1/2 complex and RUVBL1 respectively, showed the most potent inhibition in cell lines A549, H1795, HCT116, and MDA-MB-231 with IC50 values of 15 ± 1.2 µM, 15 ± 1.8 µM, 11 ± 1.0 µM, and 8.9 ± 0.9 µM respectively. A docking study of the compound was performed to predict the binding mode of pyrazolo[1,5-a]pyrimidine-3-carboxamides. Furthermore, mass spectrometry-based proteomic analysis was employed to explore cellular proteins dysregulated by treatment with compounds 16, 18, and 19. Together, the data from these analyses suggest that that compound 18 could serve as a starting point for structural modifications in order to improve potency, selectivity, and pharmacokinetic parameters of potential therapeutic molecules.
Subject(s)
Adenosine Triphosphatases , Antineoplastic Agents , ATPases Associated with Diverse Cellular Activities/metabolism , Adenosine Triphosphatases/metabolism , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Carrier Proteins/metabolism , DNA Helicases , Drug Screening Assays, Antitumor , Proteomics , Structure-Activity RelationshipABSTRACT
Enzyme replacement therapy (ERT) is a scientifically rational and clinically proven treatment for lysosomal storage diseases. Most enzymes used for ERT are purified from the culture supernatant of mammalian cells. However, it is challenging to purify lysosomal enzymes with sufficient quality and quantity for clinical use due to their low secretion levels in mammalian cell systems. To improve the secretion efficiency of recombinant lysosomal enzymes, we evaluated the impact of artificial signal peptides on the production of recombinant lysosomal enzymes in Chinese hamster ovary (CHO) cell lines. We engineered two recombinant human lysosomal enzymes, N-acetyl-α-glucosaminidase (rhNAGLU) and glucosamine (N-acetyl)-6-sulfatase (rhGNS), by replacing their native signal peptides with nine different signal peptides derived from highly secretory proteins and expressed them in CHO K1 cells. When comparing the native signal peptides, we found that rhGNS was secreted into media at higher levels than rhNAGLU. The secretion of rhNAGLU and rhGNS can, however, be carefully controlled by altering signal peptides. The secretion of rhNAGLU was relatively higher with murine Igκ light chain and human chymotrypsinogen B1 signal peptides, whereas Igκ light chain signal peptide 1 and human chymotrypsinogen B1 signal peptides were more effective for rhGNS secretion, suggesting that human chymotrypsinogen B1 signal peptide is the most appropriate for increasing lysosomal enzyme secretion. Collectively, our results indicate that altering signal peptide can modulate the secretion of recombinant lysosome enzymes and will enable lysosomal enzyme production for clinical use.
Subject(s)
Acetylglucosaminidase/metabolism , Lysosomes/enzymology , Protein Sorting Signals , Recombinant Proteins/metabolism , Sulfatases/metabolism , Acetylglucosaminidase/genetics , Animals , CHO Cells , Cricetinae , Cricetulus , Humans , Mice , Recombinant Proteins/genetics , Sulfatases/geneticsABSTRACT
OBJECTIVES: To evaluate three subtypes of continent-cutaneous urinary diversion (CCUD); Indiana pouch (IP), right colon pouch with appendico-umbilicostomy (AU), and right colon pouch with neo-appendico-umbilicostomy (NAU), by investigating diversion-specific complications and quality-of-life outcomes. MATERIALS AND METHODS: A retrospective review of an IRB-approved database was conducted for perioperative and outcome data. The EORTC QLQ-C30 questionnaire was used to assess quality of life; all responses were obtained > 6 months after diversion. RESULTS: Fifty-eight patients who underwent a CCUD at our institution from 2010 to 2016 (33 IP by two surgeons, 15 AU and ten NAU by third surgeon) were identified for this study. Higher age and Charlson Comorbidity Index (CCI) ≥ 3 were seen in the AU cohort when compared to the IP cohort (P = 0.02 and 0.02, respectively). NAU group were also older when compared to the IP group (P = 0.02). After a median follow-up of 21 months (range: 0.8-81.0), more high-grade diversion-related complications were reported for AU and NAU patients comparing to the IP group (P < 0.01 and P = 0.02, respectively). More stoma complications were also reported for the NAU cohort than the IP cohort (70% vs 30%, P = 0.03). In all groups, > 60% of stoma complications occurred at the skin or fascia level. In the 90-day postoperative period, a higher continence rate was reported for the IP cohort, and this difference was significant when compared to the NAU cohort (P = 0.04). Length of stay after surgery and revision rates were not significantly different. For all groups, the majority of patients reported little-to-no disturbance of daily functions and rated overall quality of life as good-to-excellent. CONCLUSION: Urinary diversion using the Indiana pouch and right colon pouch with appendico/neo-appendico-umbilicostomy are all associated with high rates of continence and patient satisfaction. When compared to IP, AU and NAU patients had higher rates of high-grade diversion-related complications and NAU patients had a higher stoma complications with lower 90-day continence rate.
Subject(s)
Appendix/surgery , Colon/surgery , Ostomy/adverse effects , Postoperative Complications/etiology , Quality of Life , Umbilicus/surgery , Urinary Bladder Neoplasms/surgery , Urinary Reservoirs, Continent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Retrospective Studies , Urinary DiversionABSTRACT
STUDY OBJECTIVES: To compare the prices paid for nicotine vaping products (NVPs) and supplies among current NVP users to prices paid for cigarettes among current smokers. DATA: The 2016 International Tobacco Control Four Country Vaping and Smoking Survey (4CV1). Key measures included: (1) self-reported prices paid for reusable NVPs (eg, rechargeable devices with cartridges and tank system devices with e-liquids) in the 3-month period prior to the survey among current NVP users, (2) prices paid for disposable NVPs, cartridges and e-liquids purchased in the last 30 days among current NVP users and (3) self-reported prices paid for cigarettes among current smokers. RESULTS: Disposable NVP price was higher than the price of a comparable unit for combustible cigarettes in England (EN), USA and Canada (CA). Prefilled cartridge price was higher than the price of a comparable unit of cigarettes in USA and CA, but lower in EN and Australia. E-liquid price was consistently lower than the price of a comparable unit of cigarettes across four countries. For start-up costs, price of a rechargeable device is approximately 3-5 times higher than a pack of cigarettes in four countries. CONCLUSION: NVP prices were generally higher than prices of combustible cigarettes, especially the high upfront NVP devices. The high upfront costs of purchasing a reusable NVP may discourage some smokers from switching to vaping. However, the average lower costs of cartridges and e-liquids relative to a package of cigarettes make switching to a NVP an attractive alternative to smoking in the long term so long as smokers switch completely to vaping.
Subject(s)
Electronic Nicotine Delivery Systems , Smoking Cessation , Tobacco Products , Vaping , Humans , Smokers , SmokingABSTRACT
RATIONALE: The presence of α-pyrrolidinovalerophenone (α-PVP) and its metabolites in urine is evidence of the administration of α-PVP. A toxicological challenge is that the metabolites of α-PVP exhibit amphoteric properties, which make them unsuitable for detection using gas chromatography-mass spectrometry (GC/MS). In the study reported, proper derivatization and sample extraction were essential for improving the sensitivity for GC/MS analysis. METHODS: An automated solid-phase extraction (SPE) method has been developed and optimized. The derivatization efficiency was tested using longer reaction time and the addition of polar pyridine into a mixture of N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) with 1% trimethylchlorosilane. Method validation, including linearity, limit of detection, precision, accuracy, and recovery, was evaluated using automatic SPE and GC/MS. RESULTS: The results suggested that adding pyridine to BSTFA (1:1, v/v) significantly improved derivatization efficiency and precision. After optimization, the linear range was from 25 to 1000 ng mL-1 with R2 > 0.9950. The limit of detection was 5 ng mL-1 for α-PVP and 25 ng mL-1 for OH-α-PVP. The recovery for SPE was over 88%. The inter-day and intra-day precisions were less than 15%. A forensic sample has been found containing α-PVP (67.3 ng mL-1 ) and OH-α-PVP (560.2 ng mL-1 ). CONCLUSIONS: This study is the first to validate an auto-SPE-GC/MS method for the quantification and qualification of α-PVP and OH-α-PVP in urine. We have successfully improved the derivatization efficiency and developed a sensitive and semi-automatic approach. This approach is desirable for the detection of synthetic cathinone at trace levels in biological samples.
Subject(s)
Alkaloids/urine , Gas Chromatography-Mass Spectrometry/methods , Pyrrolidines/urine , Alkaloids/metabolism , Designer Drugs/metabolism , Designer Drugs/pharmacokinetics , Humans , Limit of Detection , Pyrrolidines/metabolism , Solid Phase Extraction/methods , Substance Abuse Detection/methodsABSTRACT
Irinotecan (CPT-11), a first-line chemotherapy for advanced colorectal cancer, causes serious diarrhea in patients receiving treatment. The underlying mechanism has been shown that the active metabolite of CPT-11, SN-38, is metabolized to the inactive metabolite SN-38 glucuronide (SN-38 G) during hepatic glucuronidation, and subsequently is exported into the intestine, where SN-38 G is hydrolyzed by bacterial ß-glucuronidase (ßG) to be SN-38, thus leading to intestinal toxicity. Thus, inhibition of the intestinal bacterial ßG activity is expected to prevent CPT-11-induced diarrhea. However, the effects of such inhibition on serum pharmacokinetics of SN-38, the key determinant of CPT-11 treatment, are uncertain. Here, we determined the effects of a potent E. coli ßG (eßG)-specific inhibitor pyrazolo[4,3-c]quinoline derivative (TCH-3562) for the potential use in preventing CPT-11-induced diarrhea. TCH-3562 exhibited efficacious inhibitory potency of endogenous ßG activity in two anaerobes, Eubacteriumsp. and Peptostreptococcus anaerobius. Oral administration of TCH-3562 also effectively reduced the bacterial ßG activity in mice intestine. Moreover, pharmacokinetic analysis of TCH-3562 revealed a relatively low amount of TCH-3562 was detected in the plasma whereas the majority of TCH-3562 was found in the feces. Importantly, co-treatment of CPT-11 and TCH-3562 did not decrease active SN-38 level in mice plasma. Finally, we established that TCH-3562 as an adjuvant treatment showed protective effects on CPT-11-induced diarrhea and had no negative effects on the therapeutic efficacy of CPT-11 in tumor-bearing mice. Therefore, inhibition of the intestinal bacterial ßG activity by the specific inhibitor, TCH-3562, is promising to prevent CPT-11-induced diarrhea while maintaining its anti-tumor efficacy that may have clinical potentials for the treatment with CPT-11.
Subject(s)
Antineoplastic Agents, Phytogenic/therapeutic use , Bacterial Proteins/antagonists & inhibitors , Colonic Neoplasms/drug therapy , Diarrhea/prevention & control , Glucuronidase/antagonists & inhibitors , Irinotecan/therapeutic use , Quinolines/pharmacology , Animals , Cell Line, Tumor , Diarrhea/chemically induced , Escherichia coli/drug effects , Escherichia coli/growth & development , Eubacterium/enzymology , HEK293 Cells , Humans , Male , Mice, Inbred BALB C , Peptostreptococcus/enzymologyABSTRACT
BACKGROUND: Since Iceland became the first country to impose a ban on point-of-sale (POS) tobacco product displays in 2001, 20 countries have implemented POS display bans as of 2016. This study examined the effect that POS display bans have on smoking prevalence. METHODS: Data were sourced from Euromonitor International and the WHO MPOWER package for 2007-2014 from 77 countries worldwide. generalised linear models with country and year fixed effects were estimated to analyse the effect of POS display bans on smoking prevalence. RESULTS: Having a POS display ban reduced overall adult daily smoking, male smoking and female smoking by about 7%, 6% and 9%, respectively. CONCLUSIONS: Having a POS display ban is likely to reduce smoking prevalence and generate public health benefits.
ABSTRACT
BACKGROUND: The World Health Organization (WHO) introduced the MPOWER package to support policy implementation under the Framework Convention on Tobacco Control (FCTC). This study examined the effect of MPOWER policies on smoking prevalence and cigarette consumption in a global context. METHODS: The MPOWER composite score was constructed by adding up the six MPOWER scores for each country and survey year 2007-2008, 2010, 2012, and 2014, with a possible range between 6 (1 in each of the six score) and 29 (4 in M score and 5 in POWER scores). MPOWER composite scores that measured policy implementation were then linked to cigarette smoking prevalence and consumption data from Euromonitor International. Fractional logit and OLS regressions were employed to examine the effect of the composite MPOWER score on adult smoking prevalence and cigarette consumption, respectively. RESULTS: Results indicate that a 1-unit increase in the composite score reduces smoking prevalence by 0.2 percentage points (p<0.05) among adults and 0.3 percentage points (p<0.01) among adult males; and a reduction of 23 sticks of cigarette (1 pack of cigarettes) in cigarette consumption per capita per year. At this rate, if countries had implemented the MPOWER package to the highest levels during 2007-2014, they would have experienced a reduction in smoking prevalence of 7.26% among adults and 7.87% among adult males and a reduction of 13.80% in cigarette consumption. CONCLUSIONS: MPOWER policies were effective in reducing cigarette smoking among adults. Parties should continue to implement MPOWER policies that have been recommended by the WHO FCTC to curb tobacco epidemic.
Subject(s)
Global Health , Health Policy , Smoking Cessation/methods , Smoking/epidemiology , Adolescent , Adult , Aged , Female , Global Health/trends , Humans , Male , Middle Aged , Prevalence , Sex FactorsABSTRACT
This study investigated the effects of workplace clean indoor air law (CIAL) coverage on worksite compliance with CIALs, smoking participation among indoor workers, and secondhand smoke (SHS) exposure among nonsmoker indoor workers. This study improved on previous research by using the probability of a resident in a county covered by workplace CIALs, taking into account the state, county, and city legislation. The county-level probability of being covered by a CIAL is merged into two large nationally representative US surveys on smoking behaviors: Tobacco Use Supplement of the Current Population Survey (2001-2010) and Behavioral Risk Factor Surveillance System (2000-2006) based on the year of the survey and respondent's geographic location to identify respondents' CIAL coverage. This study estimated several model specifications of including and not including state or county fixed effects, and the effects of workplace CIALs are consistent across models. Increased coverage by workplace CIALs significantly increased likelihood of reporting a complete smoking restriction by 8% and 10% for the two different datasets, decreased smoking participation among indoor workers by 12%, and decreased SHS exposure among nonsmokers by 28%. Copyright © 2015 John Wiley & Sons, Ltd.
Subject(s)
Air Pollution, Indoor/legislation & jurisprudence , Smoking/epidemiology , Tobacco Smoke Pollution/legislation & jurisprudence , Workplace/standards , Air Pollution, Indoor/prevention & control , Humans , Smoking Prevention , United States/epidemiologyABSTRACT
Sensitive determination of the pharmacokinetics of PEGylated molecules can accelerate the process of drug development. Here, we combined different anti-PEG Fab expressing 293T cells as capture cells (293T/3.3, 293T/6.3, and 293T/15-2b cells) with four detective anti-PEG antibodies (3.3, 6.3, 7A4, or 15-2b) to optimize an anti-PEG cell-based sandwich ELISA. Then, we quantified free PEG (mPEG2K-NH2 and mPEG5K-NH2) or PEG-conjugated small molecules (mPEG5K-biotin and mPEG5K-NIR797), proteins (PegIntron and Pegasys), and nanoparticles (Liposomal-Doxorubicin and quantum-dots). The combination of 293T/15-2b cells and the 7A4 detection antibody was best sensitivity for free PEG, PEG-like molecules, and PEGylated proteins with detection at ng mL-1 levels. On the other hand, 293T/3.3 cells combined with the 15-2b antibody had the highest sensitivity for quantifying Lipo-Dox at 2 ng mL-1. All three types of anti-PEG cells combined with the 15-2b antibody had high sensitivity for quantum dot quantification down to 7 pM. These results suggest that the combination of 293T/15-2b cells and 7A4 detection antibody is the optimal pair for sensitive quantification of free PEG, PEG-like molecules, and PEGylated proteins, whereas the 293T/3.3 cells combined with 15-2b are more suitable for quantifying PEGylated nanoparticles. The optimized anti-PEG cell-based sandwich ELISA can provide a sensitive, precise, and convenient tool for the quantification of a range of PEGylated molecules.
Subject(s)
Biotin/analogs & derivatives , Immunoglobulin Fab Fragments/chemistry , Interferon-alpha/analysis , Polyethylene Glycols/analysis , Doxorubicin/analogs & derivatives , Doxorubicin/analysis , Enzyme-Linked Immunosorbent Assay/methods , HEK293 Cells , Humans , Interferon alpha-2 , Nanoparticles/analysis , Quantum Dots/analysis , Recombinant Proteins/analysisABSTRACT
OBJECTIVES: This study examines the influence that smokefree workplaces, restaurants and bars have on the adoption of smokefree rules in homes and cars, and whether there is an association with adopting smokefree rules in homes and cars. METHODS: Bivariate probit models were used to jointly estimate the likelihood of living in a smokefree home and having a smokefree car as a function of law coverage and other variables. Household data were obtained from the nationally representative Social Climate Survey of Tobacco Control 2001, 2002 and 2004-2009; clean indoor air law data were from the American Nonsmokers' Rights Foundation Tobacco Control Laws Database. RESULTS: 'Full coverage' and 'partial coverage' smokefree legislation is associated with an increased likelihood of having voluntary home and car smokefree rules compared with 'no coverage'. The association between 'full coverage' and smokefree rule in homes and cars is 5% and 4%, respectively, and the association between 'partial coverage' and smokefree rules in homes and cars is 3% and 4%, respectively. There is a positive association between the adoption of smokefree rules in homes and cars. CONCLUSIONS: Clean indoor air laws provide the additional benefit of encouraging voluntary adoption of smokefree rules in homes and cars.
Subject(s)
Air Pollution, Indoor/legislation & jurisprudence , Automobiles , Health Behavior , Housing , Smoke-Free Policy/legislation & jurisprudence , Smoking/legislation & jurisprudence , Tobacco Smoke Pollution/legislation & jurisprudence , Adolescent , Adult , Aged , Family Characteristics , Female , Humans , Male , Middle Aged , Restaurants , Smoking Prevention , Workplace , Young AdultABSTRACT
Glucuronidation is a major metabolism process of detoxification for carcinogens, 4-(methylnitrosamino)-1-(3-pyridy)-1-butanone (NNK) and 1,2-dimethylhydrazine (DMH), of reactive oxygen species (ROS). However, intestinal E. coli ß-glucuronidase (eßG) has been considered pivotal to colorectal carcinogenesis. Specific inhibition of eßG may prevent reactivating the glucuronide-carcinogen and protect the intestine from ROS-mediated carcinogenesis. In order to develop specific eßG inhibitors, we found that 59 candidate compounds obtained from the initial virtual screening had high inhibition specificity against eßG but not human ßG. In particular, we found that compounds 7145 and 4041 with naphthalenylidene-benzenesulfonamide (NYBS) are highly effective and selective to inhibit eßG activity. Compound 4041 (IC50 = 2.8 µM) shows a higher inhibiting ability than compound 7145 (IC50 = 31.6 µM) against eßG. Furthermore, the molecular docking analysis indicates that compound 4041 has two hydrophobic contacts to residues L361 and I363 in the bacterial loop, but 7145 has one contact to L361. Only compound 4041 can bind to key residue (E413) at active site of eßG via hydrogen-bonding interactions. These novel NYBS-based eßG specific inhibitors may provide as novel candidate compounds, which specifically inhibit eßG to reduce eßG-based carcinogenesis and intestinal injury.
Subject(s)
Computer Simulation , Drug Discovery/methods , Escherichia coli Proteins/antagonists & inhibitors , Glucuronidase/antagonists & inhibitors , Molecular Docking Simulation/methods , Crystallography, X-Ray , Drug Evaluation, Preclinical/methods , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/metabolism , Enzyme Inhibitors/pharmacology , Escherichia coli/drug effects , Escherichia coli/enzymology , Escherichia coli Proteins/metabolism , Gastrointestinal Microbiome/drug effects , Gastrointestinal Microbiome/physiology , Glucuronidase/chemistry , Glucuronidase/metabolism , Humans , Protein Structure, SecondaryABSTRACT
Papain-like protease (PLpro) is one of two cysteine proteases involved in the proteolytic processing of the polyproteins of Severe acute respiratory syndrome coronavirus (SARS-CoV). PLpro also shows significant in vitro deubiquitinating and de-ISGylating activities, although the detailed mechanism is still unclear. Here, the crystal structure of SARS-CoV PLpro C112S mutant in complex with ubiquitin (Ub) is reported at 1.4â Å resolution. The Ub core makes mostly hydrophilic interactions with PLpro, while the Leu-Arg-Gly-Gly C-terminus of Ub is located in the catalytic cleft of PLpro, mimicking the P4-P1 residues and providing the first atomic insights into its catalysis. One of the O atoms of the C-terminal Gly residue of Ub is located in the oxyanion hole consisting of the main-chain amides of residues 112 and 113. Mutations of residues in the PLpro-Ub interface lead to reduced catalytic activity, confirming their importance for Ub binding and/or catalysis. The structure also revealed an N-cyclohexyl-2-aminethanesulfonic acid molecule near the catalytic triad, and kinetic studies suggest that this binding site is also used by other PLpro inhibitors. Overall, the structure provides a foundation for understanding the molecular basis of coronaviral PLpro catalysis.
Subject(s)
Papain/chemistry , Severe acute respiratory syndrome-related coronavirus/chemistry , Ubiquitin/chemistry , Viral Proteins/chemistry , Amino Acid Sequence , Binding Sites , Biocatalysis , Crystallography, X-Ray , Escherichia coli/genetics , Escherichia coli/metabolism , Hydrophobic and Hydrophilic Interactions , Kinetics , Models, Molecular , Molecular Sequence Data , Mutation , Papain/genetics , Papain/metabolism , Protein Structure, Secondary , Protein Structure, Tertiary , Proteolysis , Severe acute respiratory syndrome-related coronavirus/enzymology , Sequence Alignment , Sequence Homology, Amino Acid , Substrate Specificity , Taurine/analogs & derivatives , Taurine/chemistry , Taurine/metabolism , Ubiquitin/genetics , Ubiquitin/metabolism , Viral Proteins/genetics , Viral Proteins/metabolismABSTRACT
BACKGROUNDS: A new highly pathogenic human coronavirus (CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), has emerged in Jeddah and Saudi Arabia and quickly spread to some European countries since September 2012. Until 15 May 2014, it has infected at least 572 people with a fatality rate of about 30% globally. Studies to understand the virus and to develop antiviral drugs or therapy are necessary and urgent. In the present study, MERS-CoV papain-like protease (PLpro) is expressed, and its structural and functional consequences are elucidated. RESULTS: Circular dichroism and Tyr/Trp fluorescence analyses indicated that the secondary and tertiary structure of MERS-CoV PLpro is well organized and folded. Analytical ultracentrifugation analyses demonstrated that MERS-CoV PLpro is a monomer in solution. The steady-state kinetic and deubiquitination activity assays indicated that MERS-CoV PLpro exhibits potent deubiquitination activity but lower proteolytic activity, compared with SARS-CoV PLpro. A natural mutation, Leu105, is the major reason for this difference. CONCLUSIONS: Overall, MERS-CoV PLpro bound by an endogenous metal ion shows a folded structure and potent proteolytic and deubiquitination activity. These findings provide important insights into the structural and functional properties of coronaviral PLpro family, which is applicable to develop strategies inhibiting PLpro against highly pathogenic coronaviruses.