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Tropical Medicine and Infectious Disease May 2024Diverse larval habitats significantly influence female mosquito oviposition. Utilizing traps that simulate these habitats is helpful in the study of the bioecology and...
Applicability of Traps for Collecting Mosquito Immatures (Diptera: Culicidae) for Entomological Surveillance of Arbovirus Vectors in a Remnant of the Atlantic Forest, Rio de Janeiro State, Brazil.
Diverse larval habitats significantly influence female mosquito oviposition. Utilizing traps that simulate these habitats is helpful in the study of the bioecology and characteristics of pathogen-transmitting species during oviposition. This study evaluated the feasibility of different traps in natural environments by comparing sampling methods and detecting the oviposition of epidemiologically important mosquitoes, with emphasis on species, in a fragment of the Atlantic Forest in Silva Jardim, Rio de Janeiro State, Brazil. Monthly collections were conducted from March 2021 to October 2023 using four types of traps: plastic containers, tires, bamboo, and sapucaia. Immatures were collected from these traps using a pipette, placed in plastic bags, and transported to the laboratory. Tire was the most efficient trap, showing the highest mosquito abundance (n = 1239) and number of species (S = 11). Conversely, the plastic container trap exhibited the lowest diversity (H = 0.43), with only two species and a low mosquito abundance (n = 26). The bamboo trap captured six species and recorded the second-highest diversity index (H = 1.04), while the sapucaia trap captured five species and had the third-highest diversity index (H = 0.91). Of the total immatures collected, 1817 reached adulthood, comprising 13 species, two of which are vectors of the sylvatic yellow fever virus: and . In conclusion, detecting key vectors of the sylvatic yellow fever virus in Brazil highlights the need for ongoing entomological and epidemiological surveillance in the study area and its vicinity. These efforts are crucial for monitoring vector presence and activity, identifying potential transmission hotspots, and devising effective control and prevention strategies.
PubMed: 38922037
DOI: 10.3390/tropicalmed9060125 -
Biosensors & Bioelectronics Jun 2024Automation of liquid handling is indispensable to improve throughput and reproducibility in biochemical assays. However, the incorporation of automated systems into...
Automation of liquid handling is indispensable to improve throughput and reproducibility in biochemical assays. However, the incorporation of automated systems into laboratory workflows is often hindered by the high cost and complexity associated with building robotic liquid handlers. Here, we report a 3D-printed liquid handler based on a fluidic manifold, thereby obviating the need for complex robotic mechanisms. The fluidic manifold, termed a dispensing and aspirating (DA) device, comprises parallelized multi-pipette structures connected by distribution and aspiration channels, enabling the precise supply and removal of reagents, respectively. Leveraging the versatility of 3D printing, the DA device can be custom-designed and printed to fit specific applications. As a proof-of-principle, we engineered a 3D-printed liquid handler dedicated for 3D digital rolling circle amplification (4DRCA), an advanced biochemical assay involving multiple sample preparation steps such as antibody incubation, cell fixation, nucleic acid amplification, probe hybridization, and extensive washing. We demonstrate the efficacy of the 3D-printed liquid handler to automate the preparation of clinical samples for the simultaneous, in situ analysis of oncogenic protein and transcript markers in B-cell acute lymphoblastic leukemia cells using 4DRCA. This approach provides an effective and accessible solution for liquid handling automation, offering high throughput and reproducibility in biochemical assays.
PubMed: 38905856
DOI: 10.1016/j.bios.2024.116503 -
Analytical Methods : Advancing Methods... Jun 2024A nanoporous gold microelectrode (NPG-μE) was fabricated and used for Pb(II) detection in seawater samples square wave anodic stripping voltammetry (SWASV). The Au...
A nanoporous gold microelectrode (NPG-μE) was fabricated and used for Pb(II) detection in seawater samples square wave anodic stripping voltammetry (SWASV). The Au microelectrode (Au-μE) was fabricated by embedding a gold microfiber into a Pasteur pipette, and its surface was further modified by an anodization-electrochemical reduction (A-ECR) method, yielding the NPG-μE. The fabricated electrodes were characterized by cyclic voltammetry (CV) and field emission scanning electron microscopy (FE-SEM) for electrochemical and structural morphological investigations. SWASV results show a Pb(II) stripping peak at around -0.05 V Ag/AgCl, sat. KCl, which is unusual for common Pb(II) detection (typically occurring at around -0.40 V) in anodic stripping voltammetry (ASV) analysis. The Pb(II) detection at less negative stripping potential is more beneficial. Hence, it exhibited anti-interference properties with Cd(II), which is attributed to the preferential deposition and stripping of the target analyte on the low-indexed crystal planes of the NPG structure. The calibration plot obtained by SWASV was linear in the concentration range of 0.1-10 μM, and the detection limit was found to be 57 nM (correlation coefficient of 0.9974). The NPG microsensor presented a 15-fold enhanced current response compared to Au-μE, with excellent sensitivity (27.2 μA μM cm). The application of the NPG microsensor was examined by detecting Pb(II) in seawater samples, and a satisfactory performance was obtained.
PubMed: 38904354
DOI: 10.1039/d4ay00698d -
APL Bioengineering Jun 2024Micropipette aspiration (MPA) is one of the gold standards for quantifying biological samples' mechanical properties, which are crucial from the cell membrane scale to...
Micropipette aspiration (MPA) is one of the gold standards for quantifying biological samples' mechanical properties, which are crucial from the cell membrane scale to the multicellular tissue. However, relying on the manipulation of individual home-made glass pipettes, MPA suffers from low throughput and no automation. Here, we introduce the sliding insert micropipette aspiration method, which permits parallelization and automation, thanks to the insertion of tubular pipettes, obtained by photolithography, within microfluidic channels. We show its application both at the lipid bilayer level, by probing vesicles to measure membrane bending and stretching moduli, and at the tissue level by quantifying the viscoelasticity of 3D cell aggregates. This approach opens the way to high-throughput, quantitative mechanical testing of many types of biological samples, from vesicles and individual cells to cell aggregates and explants, under dynamic physico-chemical stimuli.
PubMed: 38894959
DOI: 10.1063/5.0193333 -
Journal of Chromatography. A Jun 2024A novel fluorinated triazine-based covalent organic frameworks (F-CTFs) was designed and synthesized by using melamine and 2,3,5,6-tetrafluoroterephthalaldehydeas as...
A novel fluorinated triazine-based covalent organic frameworks (F-CTFs) was designed and synthesized by using melamine and 2,3,5,6-tetrafluoroterephthalaldehydeas as organic ligands for selective pipette tip solid-phase extraction (PT-SPE) of amphiphilic fluoroquinolones (FQs). The competitive adsorption experiment and mechanism study were carried out and verified that this F-CTFs possesses favorable adsorption affinity for FQs. The abundant fluorine affinity sites endowed the F-CTFs high selectivity to FQs extraction through F-F interactions. The adsorption capacity of F-CTFs can reach up to 109.1 mg g for enrofloxacin. The detailed characterization of the F-CTFs adsorbent involved the application of various techniques to examine its morphology and structure. Under optimized conditions, a method combining F-CTF-based PT-SPE with high-performance liquid chromatography (PT-SPE-HPLC) was established, which exhibited a broad linear range, excellent precision, and an impressively low limit of detection, and could be used for the determination of six FQs in milk, with LODs as low as 0.0010 μg mL. The recovery rates during extraction varied between 92.1% and 111.4%, exhibiting RSDs below 6.8% at different spiked concentrations.
PubMed: 38889582
DOI: 10.1016/j.chroma.2024.465078 -
Analytical Methods : Advancing Methods... Jun 2024This work describes a reliable, cheap, easy and fast method for analysis of nine bisphenols in mussel samples by gas chromatography-mass spectrometry after...
Optimization of a modified Captiva EMR-lipid method based on micro-matrix solid-phase dispersion coupled with gas chromatography-mass spectrometry for the determination of nine bisphenols in mussel samples.
This work describes a reliable, cheap, easy and fast method for analysis of nine bisphenols in mussel samples by gas chromatography-mass spectrometry after trimethylsilylation. The modified method consisted of miniaturized matrix solid phase dispersion (micro-MSPD) in a glass Pasteur pipette using Captiva EMR (enhanced matrix removal)-lipid as the sorbent. Good linearity was obtained in the work range (1-500 μg L) with a correlation coefficient () ≥ 0.998. The method accuracy and precision were determined at two concentration levels. The results show recoveries ranging from 55 to 111%. The precision varied from 1.95 to 11.4% (RSD). The whole quantification limits were between 0.056 and 3.42 μg per kg dry weight. The analytical procedure was applied for the analyses of five mussel samples collected from Galician Rias. The major compound was BPA, and wild mussels from Rías de Ferrol, Vigo and A Coruña had the highest levels. The proposed method is suitable for the analysis of BPA and its analogues in mussel samples.
Topics: Phenols; Gas Chromatography-Mass Spectrometry; Animals; Bivalvia; Benzhydryl Compounds; Limit of Detection; Lipids; Water Pollutants, Chemical
PubMed: 38847157
DOI: 10.1039/d4ay00738g -
Analytica Chimica Acta Jul 2024The convenient preparation and application of functionalized organic-inorganic hybrid monolithic materials have obtained substantial interest in the pretreatment of...
BACKGROUND
The convenient preparation and application of functionalized organic-inorganic hybrid monolithic materials have obtained substantial interest in the pretreatment of complex samples by solid-phase extraction (SPE). Compared to the in-tube solid-phase microextraction in fused-silica capillaries, micro SPE in plastic pipette tips have fascinating merits for the easily operated enrichment of trace target analytes from biological samples. However, the poor compatibility of organic-inorganic hybrid monoliths with plastics leads to the rare appearance of commercial hybrid monolithic pipette tips (HMPTs). Therefore, how to synthesize the organic-inorganic hybrid monolithic materials with better extraction performance in plastic pipette tips becomes a challenge.
RESULTS
We develop a facile and cheap strategy to immobilize organic-inorganic hybrid monoliths in pipette tips. Melamine sponge was employed as the supporting skeleton to in situ assemble amine- and thiol-bifunctionalized hybrid monolithic material via "one pot" in a pipette tip, and gold nanoparticles (GNPs) and thiol-modified aptamer against human α-thrombin were sequentially attached to the hybrid monolith within the HMPTs. The average coverage density of the aptamer with GNPs as an intermediary reached as high as 818.5 pmol μL. The enriched thrombin concentration was determined by a sensitive enzymatic chromogenic assay with the limit of detection of 2 nM. The extraction recovery of thrombin at 10 nM in human serum was 86.1 % with a relative standard deviation of 6.1 %. This proposed protocol has been applied to the enrichment and determination of thrombin in real serum sample with strong anti-interference ability, low limit of detection and high recovery.
SIGNIFICANCE
The amine- and thiol-bifunctionalized HMPTs prepared with sponge as the skeleton frame provided a novel substrate material to decorate aptamers for efficient enrichment of proteins. This enlightens us that we can take advantage of the tunability of sponge assisted HMPTs to produce and tailor a variety of micro SPE pipette tips for broader applications on the analysis of trace targets in complex biological, clinic and environmental samples.
Topics: Triazines; Aptamers, Nucleotide; Humans; Thrombin; Gold; Metal Nanoparticles; Solid Phase Extraction
PubMed: 38834272
DOI: 10.1016/j.aca.2024.342780 -
Molecular Biology of the Cell Jun 2024
Topics: Humans; Art; Science
PubMed: 38805384
DOI: 10.1091/mbc.E24-04-0149 -
BioRxiv : the Preprint Server For... May 2024Sperm cryopreservation is important for individuals undergoing infertility treatment, and for those who wish to preserve fertility potential, prior to treatments like...
Sperm cryopreservation is important for individuals undergoing infertility treatment, and for those who wish to preserve fertility potential, prior to treatments like chemotherapy, radiation therapy, gender-affirming medical interventions, elective fertility delay, or individuals in high-risk professions such as the military. Current methods for sperm cryopreservation result in approximately 30-50% decrease in sperm motility. However, recent studies have shown that ultra-rapid freezing (vitrification) is a valuable approach for maintaining sperm quality after freeze-thawing processes in the clinical laboratory setting and requires submicroliter to microliter volumes. A major challenge for the adoption of vitrification in fertility laboratories is the ability to pipette small volumes of sample. Here, we present a method that leverages open-channel droplet microfluidics to autonomously generate sub-microliter to microliter volumes of purified human sperm samples. Using a novel, open-channel droplet generator, we found no change in sperm movement and kinematic data after exposure to device and reagents in our platform. We conclude that our platform is compatible with human sperm, an important foundation for future implementation of vitrification in fertility laboratories.
PubMed: 38798664
DOI: 10.1101/2024.05.09.593416 -
Journal of Chromatography. A Jul 2024A novel azo-linked porous organic polymer (AL-POP) was synthesized from caffeic acid and benzidine via an azo-coupling reaction and characterized by FTIR, SEM-EDS, BET,...
Melamine sponges incorporated azo-linked porous organic polymer as adsorbent for extraction and determination of six B vitamins using pipette tip micro solid-phase extraction.
A novel azo-linked porous organic polymer (AL-POP) was synthesized from caffeic acid and benzidine via an azo-coupling reaction and characterized by FTIR, SEM-EDS, BET, TGA, XRD and zeta potential analysis. AL-POPs were incorporated into melamine sponges and used for pipette tip micro solid-phase extraction (PT-MSPE) of six types of B vitamins (including thiamine, riboflavin, nicotinamide, pyridoxine, folic acid, and cyanocobalamin). After extraction, the samples were analyzed using high performance liquid chromatography-diode array detection (HPLC-DAD) system. The effect of AL-POP composition on the extraction efficiency (EE) of vitamins was investigated and benzidine to caffeic acid mol ratio of 1.5, 3.35 mmol of NaNO and reaction time of 8 h were selected as optimum conditions. The efficiency of the extraction process was improved by optimizing various parameters such as the amount of sorbent, pH and ionic strength of the sample, sample volume, number of sorption and desorption cycles, type of wash solvent, and type and volume of eluent solvent. Linearity (R≥0.9987), Limit of detection (LOD) (11.88-18.97 ng/mL), limit of quantification (LOQ) (39.62-63.23 ng/mL), and enrichment factor (EF) (1.27-4.31) were obtained using calibration curves plotted under optimum conditions. Recovery values of these six B vitamins in the spiked multivitamin syrup samples varied from 80.01% to 108.35%, with a relative standard deviation (RSD) below 5.44%. Eventually, the optimized method was successfully used to extract and quantify the B vitamins in multivitamin syrup and non-alcoholic beer.
Topics: Triazines; Limit of Detection; Porosity; Chromatography, High Pressure Liquid; Vitamin B Complex; Adsorption; Polymers; Azo Compounds; Solid Phase Microextraction; Solid Phase Extraction; Hydrogen-Ion Concentration
PubMed: 38788401
DOI: 10.1016/j.chroma.2024.464978