CTBP2 contributes to cisplatin resistance in lung adenocarcinoma by inhibiting generation of reactive oxygen species.

Background: Cisplatin (CP) is commonly used for the initial treatment of lung adenocarcinoma (LUAD). Resistance to CP has long been recognized as a significant obstacle to achieving improved therapeutic outcomes. Nevertheless, the intricate molecular mechanisms underlying the phenomenon remain incompletely understood. Methods: The present study utilized the University of ALabama at Birmingham CANcer data analysis Portal (UALCAN) and Gene Expression Profiling Interactive Analysis (GEPIA) databases to conduct an analysis of the expression of C-terminal binding protein 2 (CTBP2) in LUAD. The correlation between CTBP2 expression and survival data was investigated by the Kaplan-Meier (K-M) plotter. Subsequently, the roles of CTBP2 in CP resistance were explored by analyzing cell viability, cell apoptosis, reactive oxygen species (ROS), and mitochondrial membrane potential (MMP) in CP-resistant cells (A549/DDP). Results: Our data indicated that the CTBP2 expression in LUAD exhibited a significant increase compared to the non-malignant tissues. CTBP2 overexpression showed a correlation to poor survival. CTBP2 knockdown significantly enhanced cell sensitivity to CP in A549/DDP cells. The underlying mechanism is related to promoting ROS production and decreasing MMP after CP treatment. Conclusions: CTBP2 expression has been identified as a novel biomarker for resistance to CP, and its downregulation has been found to enhance sensitivity to CP. Therefore, CTBP2 can serve as a predictor related to CP resistance and a viable therapeutic target for CP resistance in LUAD.

Constructing of efficient interface solar evaporator: In-situ colloid foaming strategy for solar desalination and visible light response sewage purification.

The shortage of drinking water has become a global problem, coastal cities can make full use of abundant seawater resources by desalination technology to ease the contradiction between supply and demand. However, fossil energy consumption contradicts the goal of reducing carbon dioxide emissions. Currently, researchers favor interfacial solar desalination devices relying only on clean solar energy. Based on the structure optimization of the evaporator, a kind of device composed of a superhydrophobic BiOI (BiOI-FD) floating layer and CuO polyurethane sponge (CuO sponge) is constructed in this paper, with its design advantages presented in the following two aspects: 1. The novel BiOI-FD photocatalyst in the floating layer reduces the surface tension and realizes the degradation of the enriched pollutants, ensuring the device to achieve solar desalination and inland sewage purification; 2. CuO sponge can inhibit salt crystallization and realize the combination of the water transport and photothermal layers. Particularly, the photothermal evaporation rate of the interface device reached 2.37 kg m-2 h-1.The novel interface evaporator design will bring a new solution for solar desalination, sewage treatment and large-scale application.

Assembly of Carbon Dots into Frameworks with Enhanced Stability and Antibacterial Activity.

Carbon dots (CDs) have been widely used as antimicrobials due to their active surface, but some CDs suffer instability. Therefore, the relative applications such as the antibacterial activity may not be reliable for long-term use. Herein, we synthesize CDs with blue fluorescence by a hydrothermal process. Thereafter, polyethylenimine was applied for the assembly of CDs into CDs-based frameworks (CDFs). The CDFs exhibited quenched fluorescence but showed more stable properties based on the scanning electron microscope and zeta potential investigations. Both CDs and CDFs show antibacterial activity toward Gram-negative Escherichia coli (E. coli) and Gram-positive Staphylococcus aureus (S. aureus), but CDFs exhibited better antibacterial performance, and S. aureus could be completely inhibited with the minimum inhibitory concentration of 30 µg/mL. This reveals CDFs magnify both the stability and antibacterial activity, which would be more promising for practical applications.

Engineering a bacterial sialyltransferase for di-sialylation of a therapeutic antibody.

Terminal α-2,6-sialylation of N-glycans is a humanized glycosylation that affects the properties and efficacy of therapeutic glycoproteins. Fc di-sialylation (a biantennary N-glycan with two α-2,6-linked sialic acids) of IgG antibodies imparts them with enhanced anti-inflammatory activity and other roles. However, the microheterogeneity of N-glycoforms presents a challenge for therapeutic development. Therefore, controlled sialylation has drawn considerable attention, but direct access to well-defined di-sialylated antibodies remains limited. Herein, a one-pot three-enzyme protocol was developed by engineering a bacterial sialyltransferase to facilitate the modification of therapeutic antibodies with N-acetylneuraminic acid or its derivatives towards optimized glycosylation. To overcome the low proficiency of bacterial sialyltransferase in antibody remodeling, the Photobacterium sp. JT-ISH-224 α-2,6-sialyltransferase (Psp2,6ST) was genetically engineered by terminal truncation and site-directed mutagenesis based on its protein crystal structure. With the optimized reaction conditions and using activity-based screening of various Psp2,6ST variants, a truncated mutant Psp2,6ST (111-511)-His6 A235M/A366G was shown to effectively improve the catalytic efficiency of antibody di-sialylation. Herceptin and the donor substrate promiscuity allow the introduction of bioorthogonal modifications of N-acetylneuraminic acid into antibodies for site-specific conjugation. 2-AB hydrophilic interaction chromatography analysis of the released N-glycans and intact mass characterization confirmed the high di-sialylation of Herceptin via the optimized one-pot three-enzyme reaction. This study established a versatile enzymatic approach for producing highly di-sialylated IgG antibodies. It provides new insights into engineering bacterial sialyltransferase for adaptation to the enzymatic glycoengineering of therapeutic antibodies and the glycosite-specific conjugation of antibodies.

Tat peptide-mediated soluble expression of the membrane protein LSECtin-CRD in Escherichia coli.

The human liver and lymph node sinusoidal endothelial cell C-type lectin (hLSECtin), a type II integral membrane protein, containing a Ca(2+)-dependent carbohydrate recognition domain (CRD), has a well-established biological activity, yet its three-dimensional structure is unknown due to low expression yields and aggregation into inclusion bodies. Previous study has demonstrated that the HIV-1 virus-encoded Tat peptide ('YGRKKRRQRRR') can increase the yields and the solubility of heterologous proteins. However, whether the Tat peptide could promote the high-yield and soluble expression of membrane proteins in Escherichia coli is not known. Therefore, the prokaryotic expression vector pET28b-Tat-hLSECtin-CRD (using pET28b and pET28b-hLSECtin-CRD as controls) was constructed, and transformed into E. coli BL21 (DE3) cells and induced with isopropyl-ß-d-thiogalactoside (IPTG) followed with identifying by SDS-PAGE and Western blot. Subsequently, the bacterial subcellular structure, in which overexpressed the heterologous proteins Tat-hLSECtin-CRD and Tat-free hLSECtin-CRD, was analyzed by transmission electron microscope (TEM) respectively, and the mannose-binding activity of Tat-hLSECtin-CRD was also determined. Expectedly, the solubility of Tat-LSECtin-CRD significantly increased compared to Tat-free LSECtin-CRD (**p < 0.01) with prolonged time, and the Tat-LSECtin-CRD had a significant mannose-binding activity. The subcellular structure analysis indicated that the bacterial cells overexpressed Tat-hLSECtin-CRD exhibited denser region compared with controls, while dot denser region aggregated in the two ends of bacterial cells overexpressed Tat-free hLSECtin-CRD. This study provided a novel method for improving the soluble expression of membrane proteins in prokaryotic systems by fusion with the Tat peptide, which may be potentially expanded to the expression of other membrane proteins.

Factors predicting inhaled corticosteroid responsiveness in African American patients with asthma.

BACKGROUND: African American patients disproportionately experience uncontrolled asthma. Treatment with an inhaled corticosteroid (ICS) is considered first-line therapy for persistent asthma. OBJECTIVE: We sought to determine the degree to which African American patients respond to ICS medication and whether the level of response is influenced by other factors, including genetic ancestry. METHODS: Patients aged 12 to 56 years who received care from a large health system in southeast Michigan and who resided in Detroit were recruited to participate if they had a diagnosis of asthma. Patients were treated with 6 weeks of inhaled beclomethasone dipropionate, and pulmonary function was remeasured after treatment. Ancestry was determined by genotyping ancestry-informative markers. The main outcome measure was ICS responsiveness defined as the change in prebronchodilator FEV(1) over the 6-week course of treatment. RESULTS: Among 147 participating African American patients with asthma, average improvement in FEV(1) after 6 weeks of ICS treatment was 11.6%. The mean proportion of African ancestry in this group was 78.4%. The degree of baseline bronchodilator reversibility was the only factor consistently associated with ICS responsiveness, as measured by both an improvement in FEV(1) and patient-reported asthma control (P = .001 and P = .021, respectively). The proportion of African ancestry was not significantly associated with ICS responsiveness. CONCLUSIONS: Although baseline pulmonary function parameters appear to be associated with the likelihood to respond to ICS treatment, the proportion of genetic African ancestry does not. This study suggests that genetic ancestry might not contribute to differences in ICS controller response among African American patients with asthma.

A cluster-randomized trial to provide clinicians inhaled corticosteroid adherence information for their patients with asthma.

BACKGROUND: Inhaled corticosteroid (ICS) nonadherence is common among patients with asthma; however, interventions to improve adherence have often been complex and not easily applied to large patient populations. OBJECTIVE: To assess the effect of supplying patient adherence information to primary care providers. METHODS: Patients and providers were members of a health system serving southeast Michigan. Providers (88 intervention; 105 control) and patients (1335 intervention; 1363 control) were randomized together by practice. Patients were age 5 to 56 years, had a diagnosis of asthma, and had existing prescriptions for ICS medication. Adherence was estimated by using prescription and fill data. Unlike clinicians in the control arm, intervention arm providers could view updated ICS adherence information on their patients via electronic prescription software, and further details on patient ICS use could be viewed by selecting that option. The primary outcome was ICS adherence in last 3 months of the study period. RESULTS: At the study end for the intention-to-treat analysis, ICS adherence was not different among patients in the intervention arm compared with those in the control arm (21.3% vs 23.3%, respectively; P = .553). However, adherence was significantly higher among patients whose clinician elected to view their detailed adherence information (35.7%) compared with both control arm patients (P = .026) and intervention arm patients whose provider did not view adherence data (P = .002). CONCLUSIONS: Overall, providing adherence information to clinicians did not improve ICS use among patients with asthma. However, patient use may improve when clinicians are sufficiently interested in adherence to view the details of this medication use.

[Anticancer effects of a novel indolinone compound in nasopharyngeal carcinoma cells].

OBJECTIVE: To develop novel targeted anticancer medicines for effective treatment of nasopharyngeal carcinoma (NPC), a prevalent malignant disease in southern China and southeast Asia. METHODS: CNE cells were treated with a novel indolinone IF239 synthesized by our research group. Cell viability was determined by the acid phosphatase assay (APA). Morphologic changes and adhesion status of CNE cells treated with IF239 were observed under a light microscope. Flow cytometry was used to analyze the cell cycle phases . Key regulating molecules in the cell cycle progression were detected by Western blotting. RESULTS: IF239 had potent cytotoxic effect on CNE cells. The possible antitumor mechanisms of IF239 involved inhibition of cell adhesion and cell cycle arrest in the G2/M phase. Moreover, G2/M arrest caused by IF239 was related to up-regulation of both cyclin B1 and the phosphorylation level of CDK1. CONCLUSION: IF239 has high anticancer activity over CNE cells, and has unique anticancer mechanisms, suggesting that IF239 has promising application potentials.

Race-ethnic differences in factors associated with inhaled steroid adherence among adults with asthma.

RATIONALE: Adherence to inhaled corticosteroid (ICS) medication is known to be low overall, but tends to be lower among African-American patients when compared with white patients. OBJECTIVES: To understand the factors that contribute to ICS adherence among African-American and white adults with asthma. METHODS: Eligible individuals had a prior diagnosis of asthma, one or more ICS prescriptions, and were members of a large health maintenance organization in southeast Michigan. Individuals were sent a survey that included questions about internal factors (e.g., patient beliefs, knowledge, and motivation) and external factors (e.g., socioeconomic status, barriers to care, social support, and stressors) potentially related to ICS adherence. Adherence was calculated using electronic prescription and fill data. Stepwise regression was used to identify factors associated with adherence before and after stratifying by race-ethnicity. MEASUREMENTS AND MAIN RESULTS: Surveys were returned by 1,006 (56.3%) of 1,787 eligible patients. Adjusting for internal factors, but not external factors, diminished the relationship between race-ethnicity and ICS adherence. Among African-American patients, readiness to take ICS medication was the only internal or external factor significantly associated with ICS adherence; it explained 5.6% of the variance in adherence. Among white patients, perceived ICS necessity, ICS knowledge, doctors being perceived as the source of asthma control, and readiness to take medication were the internal factors associated with ICS adherence; these accounted for 19.8% of the variance in adherence. CONCLUSIONS: Factors associated with ICS adherence appear to differ between African-American and white patients, suggesting that group-specific approaches are needed to improve adherence.

Improving asthma care through recertification: a cluster randomized trial.

BACKGROUND: As part of recertification, the American Board of Internal Medicine requires its diplomats to complete at least 1 practice improvement module (PIM). We assessed whether completing an asthma-specific PIM resulted in improved patient outcomes. METHODS: Practices were the unit of randomization in this cluster randomized trial. Physicians in the intervention group were asked to complete the PIM through its planning phase. The primary outcome was the dispensing of an inhaled corticosteroid (ICS) after a postintervention visit for asthma. Secondary outcomes included patient reported processes of care, asthma-related heath care use, and asthma severity. Analyses were adjusted for baseline rates at the cluster-level as well as for individual sociodemographic characteristics. RESULTS: Eight practices (19 internists) were randomized to the intervention group and 8 practices (21 internists) to the control group. For the primary outcome, ICS fill rates, patients seen by intervention group physicians were not more likely to fill an ICS prescription in the postintervention period than patients seen by control group physicians (adjusted odd ratio [AOR], 1.00; 95% confidence interval [CI], 0.64-1.56). Patients seen for asthma by intervention group physicians were less likely to receive a written action plan than patients seen by control group physicians (AOR, 0.67; 95% CI, 0.48-0.93); however, they were more likely to discuss potential asthma triggers (AOR, 1.62; 95% CI, 1.08-2.42) and had lower self-reported asthma severity measures (unadjusted P = .03). Per-protocol analysis supported the latter 2 associations. CONCLUSION: A PIM designed to improve asthma care did not improve filling of ICS prescriptions but may have lessened asthma severity through an increased discussion of asthma triggers.

Patients with asthma who do not fill their inhaled corticosteroids: a study of primary nonadherence.

BACKGROUND: Adherence to inhaled corticosteroids (ICSs) is known to be poor among patients with asthma; however, little is known about patients who do not fill their ICS prescriptions (ie, primary nonadherence). OBJECTIVE: To estimate rates of primary nonadherence and to explore associated factors. METHODS: The study population was members of a large health maintenance organization in southeast Michigan who met the following criteria: age 5 to 56 years; previous diagnosis of asthma; at least 1 electronic prescription for an ICS between February 17, 2005, and June 1, 2006; and at least 3 months follow-up after the ICS prescription. Adherence was estimated by using electronic prescription information and pharmacy claims data. Multivariable stepwise analysis was used to identify factors associated with primary nonadherence compared with adherent patients. RESULTS: One thousand sixty-four patients met the study criteria and had calculable adherence. Of these patients, 82 (8%) never filled their ICS prescription. Stepwise regression identified the following factors to be associated with an increased likelihood of primary nonadherence: younger age, female sex, African American race-ethnicity, and lower rescue medication use. Factors associated with primary nonadherence differed between race-ethnic groups. CONCLUSION: Primary nonadherence was associated with lower baseline rescue medication use, which may reflect lower perceived need for ICS therapy in patients with milder asthma. Rates of primary nonadherence and the factors which influenced this outcome differed by race-ethnicity. CLINICAL IMPLICATIONS: Understanding patient characteristics associated with primary nonadherence may be important for disease management, because many patients with asthma do not fill their ICS prescriptions.