Novel Semi-Nested Real-Time PCR Assay Leveraging Extendable Blocking Probes for Improved SHOX2 Methylation Analysis in Lung Cancer.

Lung cancer is the leading cause of cancer deaths globally, necessitating effective early detection methods. Traditional diagnostics like low-dose computed tomography (LDCT) often yield high false positive rates. SHOX2 gene methylation has emerged as a promising biomarker. This study aimed to develop and validate a novel semi-nested real-time PCR assay enhancing sensitivity and specificity for detecting SHOX2 methylation using extendable blocking probes (ExBPs). The assay integrates a semi-nested PCR approach with ExBPs, enhancing the detection of low-abundance methylated SHOX2 DNA amidst unmethylated sequences. It was tested on spiked samples with varied methylation levels and on clinical samples from lung cancer patients and individuals with benign lung conditions. The assay detected methylated SHOX2 DNA down to 0.01%. Clinical evaluations confirmed its ability to effectively differentiate between lung cancer patients and those with benign conditions, demonstrating enhanced sensitivity and specificity. The use of ExBPs minimized non-target sequence amplification, crucial for reducing false positives. The novel semi-nested real-time PCR assay offers a cost-effective, highly sensitive, and specific method for detecting SHOX2 methylation, enhancing early lung cancer detection and monitoring, particularly valuable in resource-limited settings.

Anthropogenic particle abundance and characteristics in seawater and intertidal sediments of the Tonkin Bay Coast (North Vietnam).

Microplastics (MPs, plastic items from 1 µm to 5 mm in size) are present in all environmental compartments. The evaluation of their concentration, fate, and spatial distribution is still a challenge for the scientific community. This concern is just debuting in developing countries, (i.e., Asia, South America, and Africa). This study deals with the MP contamination in the abiotic marine compartments of Northern Vietnam: seawater and intertidal sediments. Four sites located in the intertidal zone or near the coastline in Tonkin Bay, Vietnam were studied. A total of 16 samples (eight for each compartment) were collected in July 2020 (rainy season) and January 2021 (dry season). Anthropogenic particles (total observed fibers and fragments) were found at levels ranging from 3 to 303 particles/m3 in seawater and from 63 to 955 particles/kg dry weight in sediments. Most of these were fibers less than 300-µm long. Higher levels of seawater at the Nam Dinh site were found in the rainy season compared to the dry one. As the river flow was estimated six times higher during the rainy season than during the dry season, these results suggest the river discharge is a potential source of contamination for the coastal zone. The temporal variability was lower for the sediments than for the seawater, suggesting the long-term integration of the anthropogenic particles in this compartment. A small portion of sorted particles were analyzed by µFTIR (8.35%), and this sub-sample was only composed of fragments. Still, fragments were mostly composed of polypropylene (PP, 82%), polyethylene (PE, 9%), and polystyrene (PS, 9%). The fragment size was similar in the two studied compartments, but it was dependent on polymer types since PS fragments (140 ± 17 µm) were smaller than those made of PE (622 ± 123 µm) and PP (869 ± 905 µm). Future works should investigate the smallest fraction of MP (even nanoplastics) as well as find solutions in order to mitigate MP contamination in the marine environment.

Microplastics in Asian freshwater ecosystems: Current knowledge and perspectives.

Plastic pollution in freshwater ecosystems, including microplastics (MPs) smaller than 5 mm, has become an emerging global concern. Asia is considered a "hot spot" for plastic pollution due to rapid economic and demographic growth, together with rapid urbanization. Here, we provide an overview of the current knowledge on MP abundance, sources, fate, and transfer in Asian freshwater ecosystems based on publications from January 2014 to May 2021. MP contamination in freshwater compartments, including water, sediment, and biota, was found to vary strongly. In water, it ranged from 0.004 items m-3 in a moderately urbanized region to more than 500,000 items m-3 in a dumping river in a highly populated watershed. In the sediment, MP abundance ranged from 1 to more than 30,000 items kg-1 dry weight. Polyethylene (PE) and polypropylene (PP) were predominant in both water and sediment compartments. MP was detected in biota samples from all the studied species, but their abundance depended on the locations and species studied. Overall, MP characteristics (form, size, color, and polymer type) depended on sources and natural constraints (mainly hydrodynamics). This study also revealed that MP in Asian freshwater ecosystems mainly originated from domestic wastewater/runoff, followed by industrial emissions, fisheries and aquaculture wastewater. Plastic waste is not efficiently recycled or incinerated in Asia, leading to MP transfer and accumulation in the aquatic environment, and, more importantly, to ingestion by low to high trophic level organisms. This work highlights several knowledge gaps to guides future research to improve MP pollution management for the sustainable development of highly populated regions such as Asia.