Endoscopic ultrasound-guided fine-needle aspiration pancreatic adenocarcinoma samples yield adequate DNA for next-generation sequencing:A cohort analysis

BACKGROUND Genetic tests are increasingly performed for the management of unresectable pancreatic cancer.For genotyping aimed samples current guidelines recommend using core specimens,although based on moderate quality evidence.However,in clinical practice among the endoscopic ultrasound(EUS) guided tissue acquisition methods,fine needle aspiration(FNA) is the most widely performed.AIM To assess the adequacy for next generation sequencing(NGS) of the DNA yielded from EUS-FNA pancreatic adenocarcinoma(PDAC) samples.METHODS Between November 2018 and December 2021,105 patients with PDAC confirmed by EUS-FNA were included in the study at our tertiary gastroenterology center.Either 22 gauge(G) or 19G FNA needles were used.One pass was dedicated to DNA extraction.DNA concentration and purity(A260/280,A260/230) were assessed by spectrophotometry.We assessed the differences in DNA parameters according to needle size and tumor characteristics(size,location) and the adequacy of the extracted DNA for NGS(defined as A260/280 ≥ 1.7,and DNA yield:≥ 10 ng for amplicon based NGS,≥ 50 ng for whole exome sequencing [WES],≥ 100 ng for whole genome sequencing [WGS]) by analysis of variance and ttest respectively.Moreover,we compared DNA purity parameters across the different DNA yield categories.RESULTS Our cohort included 49% male patients,aged 67.02 ± 8.38 years.The 22G needle was used in 71%of the cases.The DNA parameters across our samples varied as follows:DNA yield:1289 ng(inter quartile range:534.75-3101),A260/280 = 1.85(1.79-1.86),A260/230 = 2.2(1.72-2.36).DNA yield was > 10 ng in all samples and > 100 ng in 93% of them(one sample < 50 ng).There were no significant differences in the concentration and A260/280 between samples by needle size.Needle size was the only independent predictor of A260/230 which was higher in the 22G samples(P =0.038).NGS adequacy rate was 90% for 19G samples regardless of NGS type,and for 22G samples it reached 89% for WGS adequacy and 91% for WES and amplicon based NGS.Samples with DNA yield > 100 ng had significantly higher A260/280(1.89 ± 0.32 vs 1.34 ± 0.42,P = 0.013).Tumor characteristics were not corelated with the DNA parameters.CONCLUSION EUS-FNA PDAC samples yield DNA adequate for subsequent NGS.DNA amount was similar between 22G and 19G FNA needles.DNA purity parameters may vary indirectly with needle size.

Continuous Separation of Multiple Size Microparticles using Alternating Current Dielectrophoresis in Microfluidic Device with Acupuncture Needle Electrodes

The need to continuously separate multiple microparticles is required for the recent development of lab-on-chip technology. Dielectrophoresis（DEP）-based separation device is extensively used in kinds of microfluidic applications. However, such conventional DEP-based device is relatively complicated and difficult for fabrication. A concise microfluidic device is presented for effective continuous separation of multiple size particle mixtures. A pair of acupuncture needle electrodes are creatively employed and embedded in a PDMS（poly-dimethylsiloxane） hurdle for generating non-uniform electric field thereby achieving a continuous DEP separation. The separation mechanism is that the incoming particle samples with different sizes experience different negative DEP（n DEP） forces and then they can be transported into different downstream outlets. The DEP characterizations of particles are calculated, and their trajectories are numerically predicted by considering the combined action of the incoming laminar flow and the n DEP force field for guiding the separation experiments. The device performance is verified by successfully separating a three-sized particle mixture, including polystyrene microspheres with diameters of 3 μm, 10 μm and 25 μm. The separation purity is below 70% when the flow rate ratio is less than 3.5 or more than 5.1, while the separation purity can be up to more than 90% when the flow rate ratio is between 3.5 and 5.1 and meanwhile ensure the voltage output falls in between 120 V and 150 V. Such simple DEP-based separation device has extensive applications in future microfluidic systems.

Evolution of transbronchial needle aspiration needles: Over the last half century

Transbronchial needle aspiration(TBNA)is a commonly used sampling approach in the diagnosis of hilar and mediastinal lymphadenopathy as well as peripheral lesions.As a very important tool,the continued innovation of TBNA needles is a vital driving force for the development of the technique.Although TBNA plays an important role in interventional pulmonology,there are no clear standards guiding operators to choose an appropriate needle for their operation.In recent decades,with the advent of endobronchial ultrasound-guided TBNA(EBUS-TBNA),the real-time visualization of TBNA has been enabled.These modern TBNA needles,such as ViziShot2,FLEX 19G,Acquire FNB,and EchoTip ProCore,have made significant progress in specimen collection,convenience,and safety,though still remain grounded in the basic premise and initial upgrades to the original conventional TBNA(cTBNA)needles.This review introduced the developmental history of WANG cTBNA needles,and summarized the lessons of success and failure and the enlightenments for currently used EBUS-and other emerging TBNA needles,aiming to provide a significant reference for pulmonologists who lived through the cTBNA era and for junior physicians who start working in the EBUS-TBNA era.Despite its long history,TBNA is still playing significant roles in the diagnosis of pulmonary diseases.A deeper understanding from the historical perspectives would facilitate continued innovations in the field of TBNA and beyond.