Design and Synthesis of Cyclic Dinucleotide Analogues Containing Triazolyl C-Nucleosides.

Natural cyclic dinucleotide (CDN) is the secondary messenger involved in bacterial hemostasis, human innate immunity, and bacterial antiphage immunity. Synthetic CDN and its analogues are key molecular probes and potential immunotherapeutic agents. Several CDN analogues are under clinical research for antitumor immunotherapy. A myriad of synthetic methods have been developed and reported for the preparation of CDN and its analogues. However, most of the protocols require multiple steps, and only one CDN or its analogue is prepared at a time. In this study, a strategy based on a macrocyclic ribose phosphate skeleton containing a 1'-alkynyl group was designed and developed to prepare CDN analogues containing triazolyl C-nucleosides by click chemistry. Combinatorial application of click chemistry and the sulfenylation cascade to the macrocyclic skeleton expanded the diversity of the CDN analogues. This macrocyclic skeleton strategy rapidly and efficiently provides CDN analogues to facilitate research on microbiology, immunology, and immunotherapy.

Embryo developmental toxicity in marine medaka (Oryzias melastigma) due to parental and embryonic 17α-ethinylestradiol exposure.

The synthetic estrogen 17α-ethinylestradiol (EE2) is a common component of hormone therapy and oral contraceptives and has been widely used for nearly 60 years. Numerous studies have shown that exposure to EE2 can affect embryonic development in a number of fish species. The effects of parental and embryonic EE2 exposure on embryo developmental toxicity and the underlying molecular mechanisms, however, have rarely been examined. In this study, embryos collected from parental EE2-exposed adult fish were examined to assess EE2-induecd toxicity during embryo development. The rate of embryo development including heart rate, hatching rate, and larval locomotion were measured to assess embryo developmental toxicity. The embryonic transcriptome was used to delineate the related developmental toxicity pathways. Our results suggest that parental and embryonic EE2 exposure resulted in growth retardation including a reduction in embryo heart rate, a delay in the appearance eye pigmentation, decreased hatching rate and impaired larval locomotion. In addition, gene ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, and Ingenuity Pathway Analysis (IPA) of transcriptome revealed that these impairments are controlled by estrogen receptor and related to eye structure, neuronal and synaptic structure, and behaviour. The key factors identified, including PRKAA2, APOB, EPHB2, OXTR, NR2E3, and POU4F2, could serve as biomarkers for assessing EE2-induced embryo developmental toxicity. For the first time, our results show that eye pigmentation is a potentially sensitive marker of EE2-induced embryo developmental toxicity.

A systematic study of microplastic occurrence in urban water networks of a metropolis.

The release of microplastics from sewage treatment works (STWs) into the oceans around coastal cities is well documented. However, there are fewer studies on the microplastic abundance in stormwater drains and their emissions into the coastal marine environment via sewage and stormwater drainage networks. Here, we comprehensively investigated microplastic abundance in 66 sewage and 18 sludge samples collected from different process stages at three typical STWs and 36 water samples taken from six major stormwater drains during the dry and wet seasons in Hong Kong, which is a metropolitan city in south China. The results showed that microplastics were detected in all the sewage and stormwater samples, with the abundance ranging from 0.07 to 91.9 and from 0.4 to 36.48 particles/L, respectively, and in all the sludge samples with the abundance ranging from 167 to 936 particles/g (d. w.). There were no significant seasonal variations in the microplastic abundance across all samples of sewage, sludge, and stormwater. For both waterborne sample types, a smaller size (0.02-0.3 mm) and fiber shape were the dominant characteristics of the microplastics. Polyethylene terephthalate (PET) and polypropylene (PP) were the most abundant polymer types in the sewage samples, while polyethylene (PE), PET, PP, and PE-PP copolymer were the most abundant polymer types in the stormwater samples. The estimated range of total daily microplastic loads in the effluent from STWs in Hong Kong is estimated to be 4.48 × 109 - 2.68 × 1010 particles/day, demonstrating that STWs are major pathways of microplastics in coastal environments despite the high removal percentage of microplastics in sewage treatment processes examined. This is the first comprehensive study on microplastics in the urban waters of a coastal metropolis. However, further studies on other coastal cities will enable an accurate estimation of the microplastic contribution of stormwater drains to the world's oceans.

Spatiotemporal occurrence of phthalate esters in stormwater drains of Hong Kong, China: Mass loading and source identification.

Urban stormwater is an important pathway for transporting anthropogenic pollutants to water bodies. Phthalate esters (PAEs) are endocrine disruptors owing to their estrogenic activity and potential carcinogenicity and their ubiquitous presence has garnered global interest. However, their transportation by urban stormwater has been largely overlooked. This study, for the first time, investigated 15 PAEs in stormwater from six major stormwater drains in the highly urbanized Hong Kong, a major metropolitan city in China. The results showed that PAEs were ubiquitous in the stormwater of Hong Kong, with total concentrations (∑15PAEs) spanning from 195 to 80,500 ng/L. Bis(2-n-butoxyethyl) phthalate (DBEP), diisopentyl phthalate (DiPP), dicyclohexyl phthalate (DCHP) and di-n-pentyl phthalate (DnPP) were detected in stormwater for the first time. Spatial variations in PAEs were observed among different stormwater drains, possibly due to the different land use patterns and intensities of human activities in their respective catchments. The highest and lowest levels of ∑15PAEs were found in Kwai Chung (3860 ± 1960 ng/L) and the Ng Tung River (672 ± 557 ng/L), respectively. Additionally, significantly higher concentrations of ∑15PAEs in stormwater were found in the wet season (2520 ± 2050 ng/L) than in the dry season (947 ± 904 ng/L). Principal component analysis classified domestic and industrial origins as two important sources of PAEs in the stormwater of Hong Kong. Stormwater played a crucial role in transporting PAEs, with an estimated annual flux of 0.705-29.4 kg. Thus, possible stormwater management measures were proposed to protect the receiving environment and local ecosystems from stormwater.

Microplastics: A major source of phthalate esters in aquatic environments.

Phthalate esters (PAEs) are predominant additives in plastics, their widespread contamination in aquatic environments has raised global concern. Here, twelve plastic products were prepared as microplastics to investigate their release behaviors of PAEs. Six out of 15 PAEs were quantified after 14 days of incubation in water. The leaching potentials were plastic type-specific, where the pencil case (polyvinyl chloride, PVC) represented the highest migrations with total ∑15 PAEs concentration of 6660 ± 513 ng/g, followed by the cleaning brush-1 (polyamide, PA, ~1830 ng/g) and rubber glove (1390 ± 57.5 ng/g). Conversely, the straw (polypropylene, PP), cleaning brush-2 (polyethylene terephthalate, PET) and shampoo bottle (PET) released the lowest amounts of PAEs, with 50.3 ± 8.21, 93.9 ± 91.8 and 104.35 ng/g, respectively. The release patterns of PAE congeners were polymer type-related, where di-n-butyl phthalate (DBP) dominated the leaching from PA, PP and PET microplastics (47-84%), diethyl phthalate leached the most from PVC and rubber microplastics (45-92%), while diisobutyl phthalate and DBP dominated the leaching from PE microplastics (68-94%). Water chemical properties could affect PAEs migration and the kinetic leaching process was well fitted with the pseudo-first-order model. Approximately 57.8-16,100 kg/year of PAEs were estimated to be released into oceans from microplastics.

Intra-day microplastic variations in wastewater: A case study of a sewage treatment plant in Hong Kong.

Wastewater treatment plants (WWTPs) in coastal region play a primary role in transferring microplastics into the marine environment. Wastewater is closely related to anthropogenic activities, thus the intra-day variation of abundance of microplastics in the influent should be large and could have significant impact on their estimation of the daily mass load. In this study, a 2-hour interval sampling campaign was conducted at a secondary WWTP in Hong Kong to investigate the intra-day variations and daily loads of microplastics in influent. Results show that the average microplastic abundances increased from 7.1 ± 6.0 to 12.8 ± 5.8 particles/L over time, with predominant particle sizes ranging 1-5 mm. Approximately 80% of the microplastics in samples collected from 9:30-15:00 were polyethylene and polyester, while most samples collected at 17:00 were polypropylene and polyurethane. Microplastic loads exhibited large intra-day variations ranging 6.60 × 108-1.16 × 109 particles/day, indicating that calculated daily microplastic loads based on a specific sampling period may inaccurately estimate the actual daily load.

CYP4A11 is involved in the development of nonalcoholic fatty liver disease via ROS­induced lipid peroxidation and inflammation.

Nonalcoholic fatty liver disease (NAFLD) is a fat metabolism disorder that occurs in liver cells. The development of NAFLD is considered to be associated with hepatic oxidative stress. The present study aimed to investigate the role of cytochrome P450 4A11 (CYP4A11) in the pathogenesis of NAFLD. The levels of plasma CYP4A11 and lipid peroxidation products levels exhibited a high correlation, and were increased significantly compared with those from normal subjects. Further in vitro studies demonstrated that the expression levels of CYP4A11 and the content of reactive oxygen species (ROS) were increased in free fatty acid (FFA)­stimulated HepG2 cells. Clofibrate, a CYP4A11 inducer, aggravated cell damage. Opposite results were observed for the CYP4A11 inhibitor HET0016, which attenuated apoptosis in FFA­treated cells. Furthermore, CYP4A11 gene overexpression and silencing were used to investigate the effects on inflammatory cytokine secretion. The data demonstrated that CYP4A11 promoted an increase in the mRNA expression of tumor necrosis factor α, interleukin (IL)­1ß and IL­6 in response to FFA. In addition, western blot analysis highlighted that CYP4A11 caused an upregulation of phosphorylated p65 levels and therefore affected the NF­κB signaling pathway. The data demonstrated that CYP4A11 may metabolize fatty acids to promote the production of ROS and accelerate the progression of NAFLD.

Development and Evaluation of a Water-in-oil Microemulsion Formulation for the Transdermal Drug Delivery of Teriflunomide (A771726).

Teriflunomide (TEF, A771726) is the active metabolite of leflunomide (LEF), a disease-modifying anti-rheumatic drug. The main purpose of this study was to develop and evaluate water-in-oil (W/O) microemulsion formulation of TEF. The W/O microemulsion was optimized formula is the physical and chemical stability of lecithin, ethanol, isopropyl myristate (IPM) and water (20.65/20.78/41.52/17.05 w/w) by using the pseudo-ternary phase diagram and the average droplet size is about 40 nm. The permeability of TEF microemulsion is about 6 times higher than control group in vitro penetration test. The results of anti-inflammatory effect showed that compared with the control group, the external TEF microemulsion group could significantly inhibit swelling of paw in rats, and no significant difference compared with oral LEF group. The results of hepatotoxicity test show that there were normal content of alanine aminotransferase (ALT)/aspartate aminotransferase (AST) and no obvious inflammatory infiltration of TEF microemulsion group compared with LEF group. The plasma concentration curve showed that compared with LEF group, the peak concentration of TEF microemulsion group was decreased, the half-life (t1/2) was prolonged, and the relative bioavailability of TEF microemulsion was 75.35%. These results suggest that TEF W/O microemulsion can be used as a promising preparation to play an anti-inflammatory role while significantly reducing hepatotoxicity.

Remediation of cadmium, lead and zinc in contaminated soil with CETSA and MA/AA.

Soil washing, which is used to remove heavy metals from soil, is dependent on suitable washing agents. However, there is still a lack of economical, environmentally friendly washing agents with high removal efficiency. In this study, three washing agents, carboxyalkylthiosuccinic acid (CETSA), copolymer of maleic and acrylic acid (MA/AA) and ethylenediamine tetra acetic acid (EDTA), were used to remove heavy metals from contaminated soil. The influence of washing solution concentration, pH and washing time on heavy metals removal was also investigated. The cadmium (Cd), lead (Pb), and zinc (Zn) removal efficiencies increased as washing solution concentrations increased from 0 to 60 g L-1, while they declined as pH increased from 3 to 8. Despite fluctuations between 90 and 120 min, heavy metal removal efficiencies increased continuously from 10 to 90 min. The three agents also effectively reduced the potential risks of Cd, Pb, and Zn in contaminated soil, but only CETSA and MA/AA produced no significant changes in chemical properties. Fourier transform infrared spectroscopy revealed that the hydroxyl, carboxyl, carbonyl, methoxyl, and sulfur groups were related to the heavy metal ions from the soil colloids. Thus, CETSA and MA/AA were suitable washing agents for remediation of heavy metals contaminated soil.

Feasibility of nanoscale zero-valent iron to enhance the removal efficiencies of heavy metals from polluted soils by organic acids.

Soil washing with natural chelators to remediate metal-contaminated soils has been gained attention by researchers. However, the abilities of the chelators to remediate the multiple metal polluted soils are less effective. This study employed zero-valent iron nanoparticle (nZVI) to enhance the removal efficiencies of citric (CA), tartaric (TA) and oxalic acids (OA), and evaluate their feasibility. Results showed that metal removal efficiencies increased with the increasing concentration of nZVI and soil-liquid ratio, decreased with the increasing solution pH. The kinetic simulation indicated that pseudo-first-order and pseudo-second-order models could be used for describing the washing processes. Additionally, metal removals were significantly improved by addition of nZVI (p < 0.05). The highest enhancements of soil Cd, Pb and Zn removals under solution pH of 4.0, soil-liquid ratio of 1:20 and washing time of 120 min reached 12.83% (OA- nZVI), 24.92% (CA-nZVI) and 11.64% (OA- nZVI) for mine soil, and 19.24% (TA- nZVI), 18.16% (CA-nZVI) and 8.93% (OA- nZVI) for farmland soil, respectively. After soil washing, the exchangeable forms and the environmental risks of residual metals were markedly diminished in soils. Therefore, the combinations of the organic acids and nZVI are the feasible practices to repair the soils contaminated by heavy metals.