Facile Way to Prepare a Porous Molecular Imprinting Lock for Specifically Recognizing Oxytetracyclin Based on Coordination.

Molecularly imprinted polymers (MIPs) are a kind of synthetic receptor-like materials. They have drawn more and more attention in the past decades. In this work, a facile method was developed to prepare porous magnetic MIPs utilizing metal coordination. The preparation is simply done using conventional oil-in-water emulsifier-free emulsion technology by mixing poly(styrene-co-itaconic acid), oxytetracyclin (OTC), Cu(II), and Fe3O4 magnetic fluid in one pot with a reaction time of 3 h. The product shows high specificity and selectivity toward OTC, as well as an excellent saturation adsorption capacity (62.567 mg/g). Emphasizing that the imprinting factor is 29, which is the highest one among the reported MIPs to the best of our knowledge. Combined with high-performance liquid chromatography, it was used successfully to determine OTC in pork liver, one of the most complex bio-samples. Recoveries are higher than 91.0% with relative standard deviations less than 4.5% at three spiked levels (n = 3). All evidence testifies that the MIPs based on metal coordination show excellent recognition selectivity and specificity, as well as large rebinding capacity. The strategy holds promise as a reliable, extensible, and versatile way for preparing a metal ion-mediated molecular-imprinting polymer.

Targets of biologic disease-modifying antirheumatic drugs and risk of multiple myeloma.

Several commonly used immune-suppressing biologic drugs target proteins and cytokines involved in myeloma pathogenesis. Our objective was to determine whether targeted biologic disease-modifying antirheumatic drugs (DMARDs) are associated with risk of multiple myeloma (MM). We conducted a nested case-control study within a retrospective cohort of 56,886 commercially insured adults undergoing treatment for rheumatoid arthritis, psoriatic arthritis or ankylosing spondylitis between 2009 and 2015 using the Truven Health MarketScan Databases. MM cases (n = 287) were matched to up to 10 controls (n = 2,760) on age, sex and rheumatologic indication using incidence density sampling without replacement. Our exposures of interest were biologic DMARDs targeting tumor necrosis factor-alpha, interleukin 6, cytotoxic t-lymphocyte-associated protein-4 and depletion of B cells. Relative risks were estimated as adjusted odds ratios (OR) and 95% confidence intervals (CI) using conditional logistic regression models. Cases and controls were similar with respect to use of prescription NSAIDs and concurrent conventional-synthetic DMARDs. Cases had slightly fewer etanercept users (4% vs. 7%) and slightly more tocilizumab users (1.4% vs. 0.4%). Compared to patients treated with only conventional-synthetic DMARDs, those receiving concomitant conventional-synthetic plus biologic DMARDs had lower risk of developing MM (OR = 0.48; 95% CI 0.30-0.88; p = 0.02). Risks differed by drug target with an inverse association observed with use of etanercept inhibiting tumor necrosis factor-alpha (OR = 0.55; 95% CI 0.30-1.02; p = 0.06) and a positive association with tocilizumab inhibiting interleukin-6 (OR = 4.33; 95% CI 1.33-14.19; p = 0.02) compared to biologic DMARD-naïve patients. Further investigation is warranted to understand the roles of drugs suppressing tumor necrosis factor-alpha and interleukin-6 in myeloma pathogenesis.

Myelodysplastic syndrome and acute myeloid leukemia after receipt of granulocyte colony-stimulating factors in older patients with non-Hodgkin lymphoma.

BACKGROUND: Granulocyte colony-stimulating factors (G-CSFs), which are used for the prevention of complications from chemotherapy-related neutropenia, are linked to the risk of developing second primary myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). The objective of this study was to examine the correlation between using a specific G-CSF agent and the risk of MDS/AML among older patients with non-Hodgkin lymphoma (NHL). METHODS: This was a retrospective cohort study of adults aged >65 years who were diagnosed with first primary NHL between 2001 and 2011. With data from the Surveillance, Epidemiology, and End Results-Medicare-linked database, adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) were estimated for the risk of MDS/AML associated with the receipt of G-CSF(filgrastim and pegfilgrastim) in Cox proportional-hazards models, which were stratified according to treatment accounting for confounding by indication. RESULTS: Among 18,245 patients with NHL patients who had a median follow-up of 3.5 years, 56% received chemotherapy and/or immunotherapy, and G-CSF was most commonly used in those who received rituximab plus multiple chemotherapy regimens (77%). Subsequent MDS/AML diagnoses were identified in 666 patients (3.7%). A modest increased risk of MDS/AML was observed with the receipt of G-CSF (HR, 1.28; 95% CI, 1.01-1.62) and a trend was observed with increasing doses (Ptrend < .01). When specific agents were analyzed, an increased risk of MDS/AML was consistently observed with filgrastim (≥10 doses: HR, 1.67; 95% CI, 1.25-2.23), but not with pegfilgrastim (≥10 + doses: HR, 1.11; 95% CI, 0.84-1.45). CONCLUSIONS: A higher of MDS/AML was observed in patients with NHL risk among those who received G-CSF that was specific to the use of filgrastim (≥10 doses), but not pegfilgrastim. Neutropenia prophylaxis is an essential component of highly effective NHL treatment regimens. The differential risk related to the types of G-CSF agents used warrants further study given their increasing use and newly available, US Food and Drug Administration-approved, biosimilar products.

Tumor necrosis factor-alpha inhibitors and risk of non-Hodgkin lymphoma in a cohort of adults with rheumatologic conditions.

Based on limited evidence, the U.S. Food and Drug Administration (FDA) issued a black box warning for the use of tumor necrosis factor-alpha inhibitors (TNFIs) and risk of non-Hodgkin lymphoma (NHL). Our objective was to determine the risk of NHL associated with TNFI use by duration and type of anti-TNF agent. We performed a nested case-control study within a retrospective cohort of adults with rheumatologic conditions from a U.S. commercial health insurance database between 2009 and 2015. Use of TNFIs (infliximab, adalimumab, etanercept, golimumab and certolizumab pegol) and conventional-synthetic disease-modifying antirheumatic drugs (csDMARDs) was identified, and conditional logistic regression models were used to estimate adjusted odds ratios (OR) and 95% confidence intervals (CI) for risk of NHL. From a retrospective cohort of 55,446 adult patients, 101 NHL cases and 984 controls matched on age, gender and rheumatologic indication were included. Compared to controls, NHL cases had greater TNFI use (33% vs. 20%) but were similar in csDMARD use (70% vs. 71%). TNFI ever-use was associated with nearly two-fold increased risk of NHL (OR = 1.93; 95% CI: 1.16-3.20) with suggestion of increasing risk with duration (P-trend = 0.05). TNF fusion protein (etanercept) was associated with increased NHL risk (OR = 2.73; 95% CI: 1.40-5.33), whereas risk with anti-TNF monoclonal antibodies was not statistically significant (OR = 1.77; 95% CI: 0.87-3.58). In sensitivity analyses evaluating confounding by rheumatologic disease severity, channeling bias was not likely to account for our results. Our findings support the FDA black box warning for NHL. Continued surveillance and awareness of this rare but serious adverse outcome are warranted with new TNFIs and biosimilar products forthcoming.

Effect of landfill age on the physical and chemical characteristics of waste plastics/microplastics in a waste landfill sites.

The landfills store a lot of waste plastics, thus it has been confirmed a main source for the occurrence of plastics/microplastic. Although there are some reports that microplastics (MPs) can generate in leachate and refuse samples from the landfill, it exist many blanks for the evolution of physical and chemical characteristics of waste plastics and microplastics with different landfill age. To explore the process that large pieces of plastic are fractured into microplastics, the waste plastics with landfill age from 7 to 30 years are surveyed from a typical landfill in Shanghai. The results show that PE and PP are the most common types of landfilling plastics, and their chemical composition also have changed due to the creation of CO and -OH. Moreover, the crystallinity is affected by plastic type and landfill age. The crystallinity of PP increased from 24.9% to 56.8%, but for PE, the crystallinity decreased from 55.6% to 20.8%. The mechanical properties of waste plastics were reduced significantly, which may be caused by changes in carbon-chain molecules. Al, Ti, Co, and other metal elements were detected on the plastic surface. The hydrophobic behavior of waste plastic is constantly decreasing (102.2°-80.1°) under long-term landfilling. By investigating the changes in the physical and chemical characteristics of waste plastics with different landfill age can shed light upon the process of environmental weathering of waste plastics. This provide theoretical guidance for reducing the transport of microplastics to the environment.

Copper mediated molecularly imprinted polymers for fast recognizing tylosin.

As one kind of artificial antibody, molecularly imprinted polymers (MIPs) has been widely used to separate/enrich target molecules from samples with the complex matrix prior to instrumental analysis. In this work, one novel copper mediated magnetic MIPs was developed towards anti-bacteria macrolide antibiotic tylosin (TYL). The obtained microspheres had a lot of convexities distributed evenly on the surface. And it exhibited high hydrophilicity and superparamagnetic ability, as well as excellent selectivity and specificity. Notably, it only took 5 min to reach the absorption equilibrium for TYL, which made it highly feasible for high-throughput analysis. Furthermore, the microspheres was applied as the magnetic dispersed solid phase extraction material to enrich trace TYL residue in (spiked) human urine. At three spiking levels, mean recoveries are in the range of 76.42-93.72% with relative standard deviations of 2.75-8.24% (n = 3) with HPLC/UV-Vis. The work provided one promising reference to prepare novel solid phase extraction materials for enriching/separating trace component in complex matrix.

[Preparation of molecularly imprinted polymers based on covalent organic frameworks and their application to selective recognition of trace norfloxacin in milk].

Norfloxacin (NFX) is an antibiotic that is widely used in animal husbandry. However, the presence of NFX even in trace amounts in animal-derived food may harm human health. Therefore, it is of practical significance to establish a method for monitoring NFX residues in food. Molecularly imprinted polymers (MIPs) imitate interactions established by natural receptors to selectively retain a target molecule, like antibodies or antigens do. MIPs have been widely employed in the selective recognition of specific target molecules from complex samples. Covalent organic frameworks (COFs) are a new type of organic polymer with uniform and ordered crystal structures. COFs form crystal structures by constructing organic units for ordered assembly through reversible chemical reactions. Their porous structure, regular morphology, and easy modification make COFs promising for use as excellent adsorbent carriers. Owing to these advantages of COFs, researchers have attempted to coat one MIP layer on COFs; however, the preparation methods are time-consuming and laborious, and the conditions are harsh. Hence, this study proposes a simple and rapid method for the preparation of novel MIPs with COFs as the support (DP-COF@MIPs) for the selective recognition of NFX. First, a Schiff-base COF (DP-COF) was rapidly synthesized using 3,3'-diaminobenzidine and p-phthalaldehyde with a metal trifluorate as a catalyst at room temperature. Subsequently, a two-step sequence was adopted as the synthesis strategy using NFX as the template, methacrylic acid as a functional monomer, and ethylene glycol dimethacrylate as a crosslinking agent. The entire synthesis was completed within 5 h under mild conditions. The material was then characterized by multiple analytical methods, including field-emission scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, and the Brunauer-Emmett-Teller (BET) method (to determine the specific surface area). The experimental results confirmed the successful preparation of DP-COF@MIPs. The DP-COF@MIPs presented a rough and porous surface, with a pore size of approximately 17.79 nm (mesoporous structure). The properties of the material were evaluated by adsorption and regeneration experiments. The kinetic adsorption experiment showed that the DP-COF@MIPs reached adsorption equilibrium in 90 min. Two straight lines were fitted using the pseudo-second-order kinetics model based on the experimental results for thermodynamic adsorption, indicating that the adsorption process was primarily dominated by chemical adsorption. The maximum apparent adsorption capacity was 41.57 mg/g. In the selective and competitive binding test, five drugs, namely ciprofloxacin, dimetridazole, oxytetracycline, sulfadiazine, and chloramphenicol, were selected as the interferents. The experimental results showed that the DP-COF@MIPs possessed good stereoselectivity and competitive recognition ability. The regeneration of DP-COF@MIPs was evaluated by multiple cycles of adsorption-desorption experiments. The loss in the adsorption capacity of the particles was only approximately 4.7% after seven adsorption-desorption cycles. These results from the regeneration experiments show that the DP-COF@MIPs had high stability and reusability in the selective adsorption and separation of NFX. In addition, the employed method could accurately identify trace NFX in milk samples. The average recoveries were in the range of 88.8%-92.9% at three spiked levels (0.03, 0.10, and 0.30 mg/L) with relative standard deviations (RSDs) in the range of 0.6-1.7% (n=3). Notably, the method could successfully determine NFX at contents as low as 0.0020 mg/L in the milk sample with an average recovery of 77.6% and RSD of 6.4% (n=3). This concentration is one-fiftieth of the maximum residue level stipulated by the European Union (EU), and even lower than the limit of detection (LOD) of conventional high performance liquid chromatography (HPLC) methods. The above results confirm that DP-COF@MIPs can be used to determine trace NFX in actual complex samples using HPLC equipment, even when coupled to conventional UV-Vis detectors. This study offers a facile and general method for the preparation of MIPs based on COFs with selective recognition ability.

Transcriptional regulation of the human soluble epoxide hydrolase gene EPHX2.

Soluble epoxide hydrolase (sEH) is a multifunctional protein encoded by the EPHX2 gene. The biological functions and enzyme kinetics of sEH have been extensively investigated, however, little is known about its transcriptional regulation and mechanisms of tissue specific expression. Here, a luciferase gene based reporter assay was used to identify the minimal promoter and cis regulatory elements of EPHX2. The minimal promoter was identified as a GC-rich region between nts -374 and +28 with respect to the putative transcriptional start site. A reporter plasmid carrying this minimal promoter showed higher or similar activities in comparison to a reporter plasmid carrying nts -5,974 to +28 of EPHX2 in 9 human cell lines that were tested. Sp1 binding sites that are involved in augmenting the minimal promoter activity of EPHX2 were identified by nested deletion analysis, site-specific mutation, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay.

Autophagy pathway is required for IL-6 induced neuroendocrine differentiation and chemoresistance of prostate cancer LNCaP cells.

Prostate cancer (PCa) cells undergoing neuroendocrine differentiation (NED) are clinically relevant to the development of relapsed castration-resistant PCa. Increasing evidences show that autophagy involves in the development of neuroendocrine (NE) tumors, including PCa. To clarify the effect of autophagy on NED, androgen-sensitive PCa LNCaP cells were examined. Treatment of LNCaP cells with IL-6 resulted in an induction of autophagy. In the absence of androgen, IL-6 caused an even stronger activation of autophagy. Similar result was identified in NED induction. Inhibition of autophagy with chloroquine (CQ) markedly decreased NED. This observation was confirmed by beclin1 and Atg5 silencing experiments. Further supporting the role of autophagy in NED, we found that LC3 was up-regulated in PCa tissue that had relapsed after androgen-deprivation therapy when compared with their primary tumor counterpart. LC3 staining in relapsed PCa tissue showed punctate pattern similar to the staining of chromogranin A (CgA), a marker for NED cells. Moreover, autophagy inhibition induced the apoptosis of IL-6 induced NE differentiated PCa cells. Consistently, inhibition of autophagy by knockdown of beclin1 or Atg5 sensitized NE differentiated LNCaP cells to etoposide, a chemotherapy drug. To identify the mechanisms, phosphorylation of IL-6 downstream targets was analyzed. An increase in phospho-AMPK and a decrease in phospho-mTOR were found, which implies that IL-6 regulates autophagy through the AMPK/mTOR pathway. Most important to this study is the discovery of REST, a neuronal gene-specific transcriptional repressor that is involved in autophagy activation. REST was down-regulated in IL-6 treatment. Knockdown experiments suggest that REST is critical to NED and autophagy activation by IL-6. Together, our studies imply that autophagy is involved in PCa progression and plays a cytoprotective role when NED is induced in PCa cells by IL-6 treatment. These results reveal the potential of targeting autophagy as part of a combined therapeutic regime for NE tumors.

Autophagy Blockade Sensitizes Prostate Cancer Cells towards Src Family Kinase Inhibitors.

There is overwhelming evidence that tyrosine kinases play an important role in cancer development. As a prototype of targeted therapy, tyrosine kinase inhibitors are now successfully applied to cancer treatment. However, as single agents, tyrosine kinase inhibitors have not achieved satisfactory results in the treatment of prostate cancer, principally due to their inability to efficiently kill tumor cells. The authors' laboratory has been interested in the role of the Src complex in prostate cancer progression, including the induction of androgen independence and metastasis. Previously, the authors reported that Src inhibitors such as saracatinib and PP2 caused G1 growth arrest and diminished invasiveness in prostate cancer cells but rarely apoptosis. Here, they have shown that Src family kinase (SFK) inhibitors can induce a high level of autophagy, which protects treated cells from undergoing apoptosis. Src siRNA knockdown experiments confirmed that autophagy was indeed caused by the lack of Src activity. The SFK inhibitor-induced autophagy is accompanied by the inhibition of the PI3K (type I)/Akt/mTOR signaling pathway. To test whether autophagy blockade could lead to enhanced cell death, pharmacological inhibitors (3-methyladenine and chloroquine) and a genetic inhibitor (siRNA targeting Atg7) were used in combination with SFK inhibitors. The results showed that autophagy inhibition effectively enhanced cell killing induced by SFK inhibitors. Importantly, the authors showed that a combination of saracatinib with chloroquine in mice significantly reduced prostate cancer (PC3) xenograft growth compared with the control group. Taken together, these data suggest that (1) autophagy serves a protective role in SFK inhibitor-mediated cell killing, and (2) clinically acceptable autophagy modulators may be used beneficially as adjunctive therapeutic agents for SFK inhibitors.