Typical Drugs Research Articles

Expression dynamics of periodic transcripts during cancer cell cycle progression and their correlation with anticancer drug sensitivity

BackgroundThe cell cycle is at the center of cellular activities and is orchestrated by complex regulatory mechanisms, among which transcriptional regulation is one of the most important components. Alternative splicing dramatically expands the regulatory network by producing transcript isoforms of genes to exquisitely control the cell cycle. However, the patterns of transcript isoform expression in the cell cycle are unclear. Therapies targeting cell cycle checkpoints are commonly used as anticancer therapies, but none of them have been designed or evaluated at the alternative splicing transcript level. The utility of these transcripts as markers of cell cycle-related drug sensitivity is still unknown, and studies on the expression patterns of cell cycle-targeting drug-related transcripts are also rare.MethodsTo explore alternative splicing patterns during cell cycle progression, we performed sequential transcriptomic assays following cell cycle synchronization in colon cancer HCT116 and breast cancer MDA-MB-231 cell lines, using flow cytometry and reference cell cycle transcripts to confirm the cell cycle phases of samples, and we developed a new algorithm to describe the periodic patterns of transcripts fluctuating during the cell cycle. Genomics of Drug Sensitivity in Cancer (GDSC) drug sensitivity datasets and Cancer Cell Line Encyclopedia (CCLE) transcript datasets were used to assess the correlation of genes and their transcript isoforms with drug sensitivity. We identified transcripts associated with typical drugs targeting cell cycle by determining correlation coefficients. Cytotoxicity assays were used to confirm the effect of ENST00000257904 against cyclin dependent kinase 4/6 (CDK4/6) inhibitors. Finally, alternative splicing transcripts associated with mitotic (M) phase arrest were analyzed using an RNA synthesis inhibition assay and transcriptome analysis.ResultsWe established high-resolution transcriptome datasets of synchronized cell cycle samples from colon cancer HCT116 and breast cancer MDA-MB-231 cells. The results of the cell cycle assessment showed that 43,326, 41,578 and 29,244 transcripts were found to be periodically expressed in HeLa, HCT116 and MDA-MB-231 cells, respectively, among which 1280 transcripts showed this expression pattern in all three cancer cell lines. Drug sensitivity assessments showed that a large number of these transcripts displayed a higher correlation with drug sensitivity than their corresponding genes. Cell cycle-related drug screening showed that the level of the CDK4 transcript ENST00000547281 was more significantly associated with the resistance of cells to CDK4/6 inhibitors than the level of the CDK4 reference transcript ENST00000257904. The transcriptional inhibition assay following M phase arrest further confirmed the M-phase-specific expression of the splicing transcripts. Combined with the cell cycle-related drug screening, the results also showed that a set of periodic transcripts, for example, ENST00000314392 (a dolichyl-phosphate mannosyltransferase polypeptide 2 isoform transcript), was more associated with drug sensitivity than the levels of their corresponding gene transcripts.ConclusionsIn summary, we identified a panel of cell cycle-related periodic transcripts and found that the levels of transcripts of drug target genes showed different values for predicting drug sensitivity, providing novel insights into alternative splicing-related drug development and evaluation.

Suprasomes Based on Host-Guest Molecular Recognition: An Excellent Alternative to Liposomes in Cancer Theranostics.

Liposomes and polymersomes, typical vesicular drug delivery systems (DDSs), have faced some limitations in cancer theranostics. Suprasomes, supramolecular vesicles assembled from amphiphiles linked by noncovalent interactions, show potential as new generation of vesicular DDSs. We construct suprasomes based on host-guest recognition, by which the desired functions can be integrated into carriers without tedious synthesis. Photothermally active host-guest complex is formed between a functional guest and pillar[5]arene, which further self-assembles into hollow suprasomes. A supramolecular nanomedicine is developed by encapsulating cisplatin in the suprasomes. The obtained cisplatin@Suprasomes achieve excellent anticancer efficacy and anti-metastasis combining chemotherapy and photothermal therapy, which ablate the tumors without relapse and metastasis. This work demonstrates the facile functionalization of suprasomes, holding promise as alternatives to liposomes and polymersomes.

Suprasomes Based on Host–Guest Molecular Recognition: An Excellent Alternative to Liposomes in Cancer Theranostics

AbstractLiposomes and polymersomes, typical vesicular drug delivery systems (DDSs), have faced some limitations in cancer theranostics. Suprasomes, supramolecular vesicles assembled from amphiphiles linked by noncovalent interactions, show potential as new generation of vesicular DDSs. We construct suprasomes based on host–guest recognition, by which the desired functions can be integrated into carriers without tedious synthesis. Photothermally active host–guest complex is formed between a functional guest and pillar[5]arene, which further self‐assembles into hollow suprasomes. A supramolecular nanomedicine is developed by encapsulating cisplatin in the suprasomes. The obtained cisplatin@Suprasomes achieve excellent anticancer efficacy and anti‐metastasis combining chemotherapy and photothermal therapy, which ablate the tumors without relapse and metastasis. This work demonstrates the facile functionalization of suprasomes, holding promise as alternatives to liposomes and polymersomes.

Understanding the effect of carbamazepine on the recovery of methane from lactic acid wastewater by anaerobic digestion

Anaerobic digestion is a promising technology for recovering methane from organic wastewater and contributing to carbon neutrality in wastewater plants or industrial parks. However, the interference of pharmaceuticals and personal care products with anaerobic digestion is commonly overlooked. Carbamazepine, a typical antiepileptic drug, is frequently detected in sewage sludge, but its influence on anaerobic digestion has not been systematically reported. In this work, the effect of carbamazepine on the anaerobic digestion of lactic acid synthesis wastewater was revealed comprehensively. The results showed that carbamazepine could seriously disturb anaerobic digestion performance, the methane yield decreased from 275.12 to 182.04 mL/g lactic acid as the carbamazepine dosage rose from 0 to 0.14 mM. To better understand how the drug affected anaerobic digestion, model tests were conducted on four stages of hydrolysis, acidification, acetogenesis and methanogenesis. The model tests indicated that such drug could promote the solubilization of inoculum, thereby releasing polysaccharides and proteins. But the hydrolysis stage, commonly recognized as the rate-limiting step was restricted. Meanwhile, the enhanced acidification contributed to the accumulation of volatile fatty acids. Both acetogenesis and methanogenesis were severely disturbed eventually causing a significant decrease in methane yield. The level of interference of carbamazepine on anaerobic digestion was concluded as follows: acetogenesis > methanogenesis > hydrolysis > acidification > solubilization. The high-throughput sequencing revealed the abundance of Chloroflexi increased from 21.5% to 26.7% as a main responsibility for the volatile acid accumulation. The acetoclastic methanogen Methanosaeta decreased from 59.86% to 44.78%, while the hydrogenotrophic methanogens Methanobacterium and Methanomassiliicoccus increased from 28%, 0.5%–37.9% and 4.1%, respectively. Although the recovery of methane was limited for clean production, it was more recommendable to recover the volatile fatty acids from organic wastewater or sludge with high dosage of carbamazepine.

Effectiveness and safety of Baduanjin for schizophrenia: A protocol for systematic review and meta-analysis

The cause of schizophrenia is still unknown, the course of the disease is long and its onset is thought to be related to neurodevelopmental, genetic, and oxidative stress factors and so on. There is no means of cure. Typical drug therapy is effective in treating the acute stage of schizophrenia, while the impaired social and life functions of patients are often neglected. Baduanjin is a traditional Chinese physical and breathing exercise that not only strengthens the muscles, and moves the joints, but also exercises the will. Many studies have been reported in the study on the application of Baduanjin to schizophrenic patients to promote recovery, but no research systematically evaluates the therapeutic effects and safety of Baduanjin for schizophrenic patients. This study aims to systematically investigate the efficacy and safety of Baduanjin in the treatment of schizophrenic patients. Reports of randomized controlled trials (RCTs) on Baduanjin for schizophrenia will be searched in the following data sources, including 3 English databases（PubMed, EMBASE, Cochrane Library）and 4 Chinese databases（China National Knowledge Infrastructure, Chinese Biomedical Literature, Wanfang, and China Clinical Trials Registry Database）, and their publication time is restricted from the establishment of the database to October 1, 2022. Two reviewers will independently perform study selection, data extraction, and quality assessment. RevMan V.5.4 software will be used for meta-analysis. The protocol will be performed according to preferred reporting items for systematic reviews and meta-analysis protocols (PRISMA-P) guidelines. The results will provide a systematic overview of the current evidence on the use of Baduanjin to treat schizophrenia. The conclusions of this study will help clarify whether Baduanjin is effective and safe for treating schizophrenia.

Assembly of Protein Cages for Drug Delivery.

Nanoparticles (NPs) have been widely used as target delivery vehicles for therapeutic goods; however, compared with inorganic and organic nanomaterials, protein nanomaterials have better biocompatibility and can self-assemble into highly ordered cage-like structures, which are more favorable for applications in targeted drug delivery. In this review, we concentrate on the typical protein cage nanoparticles drugs encapsulation processes, such as drug fusion expression, diffusion, electrostatic contact, covalent binding, and protein cage disassembly/recombination. The usage of protein cage nanoparticles in biomedicine is also briefly discussed. These materials can be utilized to transport small molecules, peptides, siRNA, and other medications for anti-tumor, contrast, etc.

Study on the Use of Ozone Water as a Chemical Decontamination Agent for Antineoplastic Drugs in Clinical Settings.

The exposure of healthcare workers to antineoplastic drugs in hospitals has been recognized to be harmful. To minimize the risk of exposure, the removal of these drugs from work environments, such as compounding facilities, has been recommended. In our previous paper, the degradation and inactivation efficacy of ozone water, which is being introduced into Japanese hospitals as a chemical decontamination agent, was reported for its effects on typical antineoplastic drugs (gemcitabine, irinotecan, paclitaxel). This article aims to further investigate the efficacy of ozone water for eight antineoplastic drugs to clarify its application limitations. A small amount (medicinal ingredient: typically ca. 1.5 μmol) of formulation containing 5-fluorouracil, pemetrexed, cisplatin, oxaliplatin, cyclophosphamide, ifosfamide, doxorubicin, or docetaxel was mixed with 50 mL of ozone water (~8 mg/L), and the resulting solutions were analyzed by high-performance liquid chromatography over time to observe the degradation. Consequently, the ozonation was overall effective for the degradation of the drugs, however this varied depending on the chemical structures of the drugs and additives in their formulations. In addition, after the parent drugs were completely degraded by the ozonation, the degradation mixtures were subjected to 1H nuclear magnetic resonance spectroscopy and evaluated for mutagenicity against Salmonella typhimurium strains and cytotoxicity against human cancer cells. The degradation mixtures of cisplatin and ifosfamide were mutagenic while those of the other drugs were non-mutagenic. Further, the ozonation resulted in clear decreases of cytotoxicity for 5-fluorouracil, oxaliplatin, and doxorubicin, but increases of cytotoxicity for pemetrexed, cisplatin, cyclophosphamide, and ifosfamide. These results suggest that the ozone water should be restrictedly used according to the situation of contamination in clinical settings because the ozonation enhances toxicity depending on the drug even if degradation is achieved.

Optimal exposure to docetaxel in adjuvant chemotherapy for early-stage breast cancer.

Drug-induced neutropenia is the main reason for the dose limitation of docetaxel in patients with breast cancer. The area under the drug concentration-time curve (AUC) of docetaxel is associated with neutropenia. However, the optimal exposure to docetaxel for receiving postoperative adjuvant chemotherapy remains unclear. Therefore, we aimed to evaluate the relationship between the docetaxel AUC and neutropenia, identify potential influencing factors, and explore the best monitoring target for docetaxel when treating patients with early-stage breast cancer using a population pharmacokinetic (PopPK) model. Docetaxel plasma concentration, demographics, clinical data, and related laboratory data were collected. PopPK analyses were performed using a nonlinear mixed-effect modelling program. The docetaxel AUC was determined using the maximum a posteriori Bayesian (MAPB) method. The docetaxel exposure-toxicity threshold measured from the AUC for neutropenia was determined using the receiver operating characteristic (ROC) curve. The correlation between docetaxel exposure and neutropenia was analysed using multivariable logistic regression. Among the 70 participants, 47 (67.1%) developed severe neutropenia. The PopPK analysis showed that the typical drug clearance (CL) rate was 37.4L/h. Age was a significant covariate of CL rate, and aspartate aminotransferase and albumin levels were covariables of the volume of distribution. The multivariable regression analysis showed that AUC >3.0mg.h/L (odds ratio [OR], 5.940; 95% confidence interval [CI], 1.693-20.843; P=0.005), platinum use (OR, 0.156; 95% CI, 0.043-0.562; P=0.005) and baseline haemoglobin level (OR, 0.938; 95% CI, 0.887-0.993; P=0.027) were significant factors influencing the occurrence of grade 3/4 neutropenia. The AUC of first cycle may not predict the occurrence rates of grade 3/4 neutropenia in later cycles. We developed a docetaxel PopPK model for patients with early-stage breast cancer. Age and AST and ALB levels were significant covariates. AUC estimated using the MAPB method can predict the toxicity of docetaxel in patients with breast cancer. Docetaxel AUC >3.0mg.h/L, absence of platinum use and low baseline haemoglobin level were risk factors for docetaxel-induced grade 3/4 neutropenia. Chinese Clinical Trial Center Registry (ChiCTR2200056460).

Enhancing oral bioavailability of andrographolide via sodium dodecyl sulfate and D-α-Tocopherol polyethylene glycol 1000 succinate copolymer modified nanocrystals

Nanocrystal technology is a common strategy to enhance the oral bioavailability of poorly-soluble drugs, and how to enlarge the enhancing ability on oral absorption via nanocrystals has become a pharmaceutical research field. In this study, we selected andrographolide (ADR), a typical poorly-soluble drug, as the model drug, and employed sodium dodecyl sulfate and D-ɑ-Tocopherol polyethylene glycol 1000 succinate copolymer (SDS-TPGS) to construct novel SDS-TPGS modified andrographolide nanocrystals (S-TANs). We found that the particle sizes of SDS/TPGS physical mixture modified nanocrystals (STANs) were about 553.4 ± 4.1 nm, while that of SDS-TPGS modified nanocrystals (S-TANs) were 604.6 ± 5.7 nm. During the production process of nanocrystals, the crystallinity of ADR was slightly decreased. Nanocrystals enhanced the saturation solubility of ADR, and improved oral absorption, Cmax and AUC0-∞ of S-TANs was about 2.10 and 1.24 fold increased, STAN about 2.63, 2.97 fold. In a Real-time distribution study, we found the distribution time of andrographolide, SDS and TPGS in SDS/TPGS physical mixture modified nanocrystals system varies widely, while the distribution times of these materials were similar in SDS-TPGS copolymer modified nanocrystals system. These results indicated that modified with functional stabilizer was an efficient strategy to enlarge the enhancing ability on oral absorption of nanocrystals, and synthesized copolymer containing different functional stabilizers showed higher ability on enhancing oral bioavailability than physical mixtures.

OUTCOMES OF SINGLE BAG PROTOCOLS FOR DESENSITIZATION TO ANTINEOPLASTIC AGENTS

<h3>Introduction</h3> The typical drug desensitization process includes three separate intravenous infusions with each bag containing an increasing concentration of the drug. Many biologics and chemotherapy agents may not be diluted past a specific concentration or diluted protocols are prohibitively long, making a single bag desensitization protocol useful. The aim of this case series is to describe the safety and efficacy of single bag desensitization protocols in two oncology patients. <h3>Case Description</h3> Patient A is a 55 year-old male with metastatic prostate cancer who required desensitization to Carbazitaxel following an IgE mediated reaction to another member of the taxane drug class. Patient B is a 89 year old female with metastatic squamous cell carcinoma of the tongue who required desensitization to Ceftuximab following grade 3 cyctokine release syndrome. Both patients tolerated single bag desenstization protocols in the MICU without adverse events. Both patients went on to tolerate readministration of their culprit agents in outpatient infusion centers. <h3>Discussion</h3> Developments in medical oncology are providing an increasing number of treatment modalities, making drug reactions to antineoplastic agents more common. Patients may have only one viable treatment option, necessitating desensitization following an adverse reaction. Single bag desensitization protocols may be undiluted or have a single dilution. This method should be considered as it can be well tolerated in both IgE mediated and cytokine mediated drug reactions. Single bag protocols can simplify the procedure and decrease the time required to reach the target dose, aiding transition to outpatient infusion centers thus reducing costs and increasing patient's quality of life.

Drugging p53 in cancer: one protein, many targets.

Mutations in the TP53 tumour suppressor gene are very frequent in cancer, and attempts to restore the functionality of p53 in tumours as a therapeutic strategy began decades ago. However, very few of these drug development programmes have reached late-stage clinical trials, and no p53-based therapeutics have been approved in the USA or Europe so far. This is probably because, as a nuclear transcription factor, p53 does not possess typical drug target features and has therefore long been considered undruggable. Nevertheless, several promising approaches towards p53-based therapy have emerged in recent years, including improved versions of earlier strategies and novel approaches to make undruggable targets druggable. Small molecules that can either protect p53 from its negative regulators or restore the functionality of mutant p53 proteins are gaining interest, and drugs tailored to specific types of p53 mutants are emerging. In parallel, there is renewed interest in gene therapy strategies and p53-based immunotherapy approaches. However, major concerns still remain to be addressed. This Review re-evaluates the efforts made towards targeting p53-dysfunctional cancers, and discusses the challenges encountered during clinical development.

Pharmacological investigations of effort-based decision-making in humans: Naltrexone and nicotine.

Many mental health disorders are characterized by an impaired ability, or willingness, to exert effort to obtain rewards. This impairment is modeled in effort-based decision tasks, and neuropharmacological studies implicate dopamine in this process. However, other transmitter systems such as opioidergic and cholinergic systems have received less attention. Here, in two separate studies we tested the acute effects of naltrexone and nicotine on effort-based decision-making in healthy adults. In Study 1, we compared naltrexone (50mg and 25mg) to placebo, and in Study 2, a pilot study, we compared nicotine (7mg) to placebo. In both studies, participants completed the Effort Expenditure for Rewards Task (EEfRT), which measured effort-based decision-making related to monetary rewards. Although subjects expended greater effort for larger reward magnitude and when there was a higher probability of receiving the reward, neither naltrexone nor nicotine affected willingness to exert effort for monetary rewards. Although the drugs produced significant and typical drug effects on measures of mood and behavior, they did not alter effort-based decision-making. This has implications both for the clinical use of these drugs, as well as for understanding the neuropharmacology of effort-related behavior.

Prognostic impact of suboptimal adherence to statin therapy after STEMI: results from the FAST-STEMI registry

Abstract Introduction Patterns and prognostic impact of suboptimal adherence to statin therapy, after STEMI, remains understudied Purpose To identify predictors and to estimate the prognostic impact of low statin adherence after STEMI Method We evaluated real-world purchase of statins and typical cardiovascular drugs, after STEMI, in a large regional database that enrolled patients from 2012 to 2017; adherence was defined as the ratio of bought tablets divided by the number of expected tablets in the follow-up period. Cox regression was used for multivariate analysis; Kaplan Meier and Cox proportional hazard models were performed to evaluate cumulative event rates of all-cause mortality at follow-up Results A total of 4062 patients were enrolled, of which 12.6% died after a median of 4.7 years of follow up (IQR 3.7–5.7). Statin adherence lower than 80%, identified as the best cutoff at Youden's analysis, was identified in 42% of the registry population; statin adherent patients were more likely younger, with known dyslipidemia at enrollment, discharged with ACE inhibitors and a potent P2Y12 inhibitor (ticagrelor or prasugrel), and with a statin diverse from atorvastatin. After multivariate adjustment for in-study outcome predictors, low statin adherence was independently associated with all cause mortality (HR 0.57; 95% CI 0.46–0.70; p&amp;lt;0.001), along with older age, not known dyslipidemia at enrollment and clopidogrel at discharge after STEMI Conclusions In the contemporary real-world setting, statin adherence following STEMI remains poor, especially in high risk patients, and is still a crucial independent predictor of all-cause mortality. Effective strategies to improve statint adherence remain an unmet clinical need. Funding Acknowledgement Type of funding sources: None.

S1978 A Rare Case of Leflunomide-Induced Colitis: Clinical, Endoscopic and Microscopic Findings

Introduction: Leflunomide-induced colitis is a rare entity, which can be difficult to diagnose. We present a case of an elderly woman with rheumatoid arthritis (RA) who complained of subacute diarrhea. Colonoscopy with biopsy was performed and investigation of inflammatory and infectious causes were negative. Cessation of leflunomide resolved diarrhea. A wide range of endoscopic findings and late-onset of symptoms makes this a challenging diagnosis. Our hope is to aid in the recognition of this drug-induced colitis to reduce hospitalization stays and improve treatment. Case Description/Methods: A 79-year-old female with a past medical history of RA, COPD and gout, presented to the hospital with 7 days of watery diarrhea. She reported more than 6 bowel movements daily. She denied recent sick contacts, travel, or changes in diet. Patient did not have a family history of inflammatory bowel disease or colorectal cancer. Vitals were stable. CT abdomen/pelvis demonstrated continuous wall thickening of the splenic flexure to rectum. Labs included CRP at 40.7, ESR at 58, and a positive fecal lactoferrin. Patient was empirically treated for infectious colitis with ceftriaxone and metronidazole. Stool cultures and C. difficile toxin were negative. Colonoscopy showed areas of congested and erythematous mucosa from the rectum to the transverse colon. Biopsies demonstrated increased lymphocytic infiltration of the lamina propria, thickened epithelial layer and crypts with increased intraepithelial lymphocytes. Upon further questioning, the patient reported starting leflunomide 2 months prior. She was asked to trial cessation of leflunomide upon discharge. Upon 2 week follow up, diarrhea had resolved. Discussion: Leflunomide is a disease modifying drug used to treat RA among other types of inflammatory arthritis. In approximately 20% of patients, adverse gastrointestinal side effects of abdominal pain, diarrhea and nausea are well recognized. However, drug-induced colitis has only been described in a few case reports, and even fewer endoscopic descriptions exist. Paucity of this diagnosis may be attributed to its nebulous presentation. Unlike most typical drug reactions, symptom onset can range from 18-24 months. As our case exemplifies, this can lead to unnecessary testing and prolonged hospital stays. Due to this patient's symptom improvement with supportive care, we opted for drug cessation. However, for severe colitis, steroids, cholestyramine wash, and biologics may be considered.

Chlorination of methotrexate in water revisited: Deciphering the kinetics, novel reaction mechanisms, and unexpected microbial risks

Chlorination of a typical anticancer drug with annually ascending use and global prevalence (methotrexate, MTX) in water has been studied. In addition to the analysis of kinetics in different water/wastewater matrices, high-resolution product identification and in-depth secondary risk evaluation, which were eagerly urged in the literature, were performed. It was found that the oxidation of MTX by free available chlorine (FAC) followed first-order kinetics with respect to FAC and first-order kinetics with respect to MTX. The pH-dependent rate constants (kapp) ranged from 170.00 M−1 s−1 (pH 5.0) to 2.68 M−1 s−1 (pH 9.0). The moiety-specific kinetic analysis suggested that 6 model substructures of MTX exhibited similar reactivity to the parent compound at pH 7.0. The presence of Br− greatly promoted MTX chlorination at pH 5.0–9.0, which may be ascribed to the formation of bromine with higher reactivity than FAC. Comparatively, coexisting I− or humic acid inhibited the degradation of MTX by FAC. Notably, chlorination effectively abated MTX in different real water matrices. The liquid chromatography-high resolution mass spectrometry analysis of multiple matrix-mediated chlorinated samples indicated the generation of nine transformation products (TPs) of MTX, among which seven were identified during FAC oxidation for the first time. In addition to the reported electrophilic chlorination of MTX (the major and dominant reaction pathway), the initial attacks on the amide and tertiary amine moieties with C-N bond cleavage constitute novel reaction mechanisms. No genotoxicity was observed for MTX or chlorinated solutions thereof, whereas some TPs were estimated to show multi-endpoint aquatic toxicity and higher biodegradation recalcitrance than MTX. The chlorinated mixtures of MTX with or without Br− showed a significant ability to increase the conjugative transfer frequency of plasmid-carried antibiotic resistance genes within bacteria. Overall, this work thoroughly examines the reaction kinetics together with the matrix effects, transformation mechanisms, and secondary environmental risks of MTX chlorination.

Dual effect of amitriptyline in the control of vascular tone: Direct blockade of calcium channel in smooth muscle cells and reduction of TLR4-dependent NO production in endothelial cells

Background and purposeAmitriptyline (AM) is a classical and typical tricyclic antidepressant drug. Despite its well-known effects on the nervous system, it has been described to work as a TLR4 antagonist and several clinical works suggested some unexpected cardiovascular effects. The role of amitriptyline on vascular tone is not clear, thus we hypothesized that amitriptyline has a double effect on vascular tone by both endothelial TLR4-dependent nitric oxide down-regulation and calcium channel blockade in smooth muscle cells. Experimental approachChanges in isometric tension were recorded on a wire myograph. NO production was evaluated by fluorescence microscopy and flow cytometry in the mouse aorta and EAhy926 cells using DAF fluorescence intensity. Calcium influx was evaluated in A7r5 cells by flow cytometry. Western blot was used to analyze eNOS and nNOS phosphorylation. Key resultsAM reduced PE-induced contraction by calcium influx diminution in smooth muscle cells (F/F0 = 225.6 ± 15.9 and 118.6 ± 17.6 to CT and AM, respectively). AM impaired Ach-dependent vasodilation (Emax = 95.8 ± 1.4; 78.1 ± 1.8; 60.4 ± 2.9 and −7.4 ± 1.0 for CT, 0.01, 0,1 and 1 μmol/L AM, respectively) through reduction of calcium influx and NO availability and TLR4 antagonism in a concentration-dependent manner. AM or TLR4 gene deletion significantly reduced NO production (Fluorescence = 9503 ± 871.7, 2561 ± 282, 4771 ± 728 and 1029 ± 103 to CT, AM, TLR4−/− and AM + TLR4−/−, respectively) by an increase in nNOSser852 and reduction in eNOSser1177 phosphorylation in endothelial cells. Conclusions and implicationsOur data show that amitriptyline impaired vascular function through two different mechanisms: blockade of TLR4 in endothelial cells and consequent decrease in NO production and calcium influx reduction in smooth muscle and endothelial cells. We also suggest, for the first time, nNOS activity reduction by AM in non-neuronal cells.

The orphan receptor GPR88 controls impulsivity and is a risk factor for Attention-Deficit/Hyperactivity Disorder.

The neural orphan G protein coupled receptor GPR88 is predominant in the striatum and cortex of both rodents and humans, and considered a potential target for brain disorders. Previous studies have shown multiple behavioral phenotypes in Gpr88 knockout mice, and human genetic studies have reported association with psychosis. Here we tested the possibility that GPR88 contributes to Attention Deficit Hyperactivity Disorder (ADHD). In the mouse, we tested Gpr88 knockout mice in three behavioral paradigms, best translatable between rodents and humans, and found higher motor impulsivity and reduced attention together with the reported hyperactivity. Atomoxetine, a typical ADHD drug, reduced impulsivity in mutant mice. Conditional Gpr88 knockout mice in either D1R-type or D2R-type medium spiny neurons revealed distinct implications of the two receptor populations in waiting and stopping impulsivity. Thus, animal data demonstrate that deficient GPR88 activity causally promotes ADHD-like behaviors, and identify circuit mechanisms underlying GPR88-regulated impulsivity. In humans, we performed a family-based genetic study including 567 nuclear families with DSM-IV diagnosis of ADHD. There was a minor association for SNP rs2036212 with diagnosis, treatment response and cognition. A stronger association was found for SNP rs2809817 upon patient stratification, suggesting that the T allele is a risk factor when prenatal stress is involved. Human data therefore identify GPR88 variants associated with the disease, and highlight a potential role of life trajectories to modulate GPR88 function. Overall, animal and human data concur to suggest that GPR88 signaling should be considered a key factor for diagnostic and treatment of ADHD.

Transcriptomic responses to cytotoxic drug cisplatin in water flea Daphnia magna

Cytotoxic drugs have been recognized by the European Union as the potential threat in the aquatic environment. As a typical cytotoxic drug, effects of long-term exposure to cisplatin at the environmentally relevant concentrations on the crustacean health and its molecular mechanism remain undetermined. In this study, the growth and reproduction of Daphnia magna resulting from cisplatin exposure were initially assessed. While the phenotypes were not altered in 2 μg L−1, 20 μg L−1, and 200 μg L−1 treatment groups, cisplatin at 500 µg L−1 significantly reduced the offspring number to 8–13 neonates in each brood, which was lower than 13–27 neonates in the control group. In addition to the delay in the time of first pregnancy, the body length was decreased by approximate 12.13% at day 7. Meanwhile, all daphnids died after exposure to 500 µg L−1 cisplatin for 17 days. Transcriptome profiling bioassays were performed for 10 days to explore the alternation at the molecular level. Briefly, 980 (257 up- and 723 down-regulated), 429 (182 up- and 247 down-regulated) and 1984 (616 up-regulated and 1368 down-regulated) genes were differentially expressed (adj p < 0.05) in low (2 μg L−1), medium (200 μg L−1) and high (500 μg L−1) cisplatin treatment groups, respectively. Differentially expressed genes were primarily enriched in the digestion and absorption, nerve conduction, endocrine interference, and circulatory related pathways. Specifically, the down-regulated digestive secretion and nutrient absorption and neuronal conduction pathways may lead to insufficient energy supply involved in growth and reproduction, and hinder ovarian development and cell growth. Down-regulation of ovarian steroids and relaxin signaling pathways may be related to the reduction of offspring number and delayed pregnancy, and reduced body length of D. magna may attribute to the enrichment of insulin secretion pathway. In addition, the death of D. magna may result from the reduced expression of genes in cardiomyocyte contraction and apoptosome processes. Taken together, this study revealed the potential toxic mechanism of cisplatin in a model water flea.

Multistage defense response of microalgae exposed to pharmaceuticals in wastewater

Recently, the use of microalgae for bioremediation of pharmaceuticals (PhAs) has attracted increasing interest. However, most studies focused more on microalgae removal performance, its defensive response to the PhAs during wastewater treatment remains unexplored. Herein, microalgal three defensive systems have been investigated in synthetic wastewater, with six PhAs as the typical drug. Results show that PhAs could bind to EPS, and this action in turn could help to alleviate the direct toxicity of PhAs to microalgae. Subsequently, the physiological analyses revealed the increase of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities, potentially reducing the oxidative stress induced by PhAs. Furthermore, the enzyme activities of cytochrome P450 (CYP450) and glutathione-S-transferase (GST) were significantly upregulated after exposure to SMX, CIP and BPA, followed by a significant decrease in biodegradation rates after the addition of CYP450 inhibitors, suggesting that the biotransformation and detoxification of PhAs occurred. Meanwhile, molecular docking further revealed that CYP450 could bind with PhAs via hydrogen bond and hydrophobic interaction, which proved their abilities to be metabolized and form transformation products in microalgae. These findings provide an advancing understanding of microalgae technologies to improve the treatment of wastewater contaminated with PhAs.

Prescription Advice Based on Data of Drug-Drug-Gene Interaction of Patients with Polypharmacy.

PurposePharmacogenetic counselling is a complex task and requires the efforts of an interdisciplinary team, which cannot be implemented in most cases. Therefore, simple rules could help to minimize the risk of medications incompatible with each other or with frequent genetic variants.Patients and MethodsOne hundred and eighty-four multi-morbid Caucasian patients suffering from side effects or inefficient therapy were enrolled and genotyped. Their medication was analyzed by a team of specialists using Drug-PIN® (medication support system) and individual recommendations for 34 drug classes were generated.ResultsIn each of the critical drug classes, 50% of the drugs cannot be recommended to be prescribed in typical drug cocktails. PPIs and SSRI/SNRIs represent the most critical drug classes without showing a single favorable drug. Among the well-tolerated drugs (not recommended for less than 5% of the patients) are metamizole, celecoxib, olmesartan and famotidine. For each drug class, a ranking of active ingredients according to their suitability is presented.ConclusionGenotyping and its profound analysis are not available in many settings today. The consideration of frequent alterations of metabolic elimination routes and drug–drug–gene interactions by using simple rankings can help to avoid many incompatibilities, side effects and inefficient therapies.