Sample Loading Research Articles

Acute Human Bocavirus 1 Infection in Children Hospitalized for Acute Bronchiolitis: A 2-Year Prospective Study.

The objective of this prospective study was to assess the proportion and clinical consequences of human bocavirus 1 (HBoV1) replication in children hospitalized for acute bronchiolitis (AB) with HBoV1 DNA in the nasopharynx (NP). For this purpose, we detected HBoV1 DNA and mRNA (evidence of viral replication and viable virus) in NP in cases and healthy control children. This research allowed us to distinguish active HBoV1 infections from inactive ones. HBoV1 DNA was detected in the NP of 37 of 307 patients with AB (12.1%) and 9 of 150 children in a healthy control group (6%) with a high codetection rate with other respiratory viruses in AB patients, 28 of 37 (75.7%). Only 9 of 37 HBoV1 DNA-positive NP swabs (24.3%) with high DNA load were also HBoV1 mRNA positive, moreover, HBoV1 DNA was also detected in the plasma of these patients. Based on the results of our study, we can conclude that children with AB acute HBoV1 infection has a high HBoV1 DNA load in NP samples together with detected HBoV1 mRNA and detected HBoV1 DNA in plasma.

TPE-xgboost for explainable predictions of concrete compressive strength considering compositions, and mechanical and microstructure properties of testing samples

The use of machine learning (ML) for predicting concrete compressive strength (CCS) has shown promising and accurate results, making it a valuable tool in the field. However, efficient prediction requires not only a robust ML approach but also a comprehensive and well-curated dataset. This study addresses this challenge by investigating an extensive and integrated dataset of (1) concrete compositions (mixture proportions) and (2) testing conditions (mechanical and microstructure properties of concrete testing samples), containing 1525 observations relating to 39 parameters. On algorithms side, this research proposes novel tree-structured parzen estimator based extreme gradient boosting (TPE-xgboost) for accurate and confident CCS predictions. Moreover, SHapley Additive exPlanations (SHAP) analysis was performed to provide a comprehensive understanding of feature importance, dependencies, and interactions. Compared to prior research, this study demonstrates significant improvements in CCS prediction accuracy for separate investigations on concrete compositions (3.77%) and mechanical and microstructure properties (28.57%), while maintaining reasonable accuracy for the integrated dataset despite its complexity and sparseness. SHAP analysis reveals key influential factors, including age and water-to-binder ratio in concrete composition, and peak load and height of testing samples, as well as microstructural characteristics such as mean values of global autocorrelation length and its integral range. This research provides valuable insights into model optimization, predictive performance, and feature dependencies and interactions, contributing to the development of more accurate and reliable CCS prediction models.

Effect of ozone gassing time and stirring duration on peroxide value, microbial decontamination, aflatoxin detoxification and sensorial characteristics of dried pistachios

Aflatoxin contamination in agricultural products such as pistachios threatens human health. Ozone has intense antimicrobial activity against microorganisms. The effect of ozone-treated time (3,4 and 5 hours) and stirring time (10, 15 and 20 minutes) on microbial, aflatoxin content, peroxide value, and sensory properties of aflatoxin contaminated pistachios were investigated in this study. The microbial test results also show a significant reduction of microbial load in ozone-treated samples. According to the results, ozone gas caused a significant reduction in aflatoxin levels in pistachio kernels. Aflatoxins B1, B2, G1, G2 and total which was 33.5, 1.5, 0.06, 0. 31 and 35.13 ppb in the control sample, reached 0 ppb after 5 hours of ozone-treatment (15 or 20 min). The results show that the longer the ozone gas is gasified, the higher the peroxide value. The lowest and highest peroxide value was observed in the control and the ozone-treated sample for 5 hours (stirring the samples every 20 minutes), respectively. All peroxide values were below the allowable limit (1 meq/kg). The results of sensory evaluation show that ozonation did not have a negative effect on the falvor, texture, and color score of pistachios, but only had a significant effect on the aroma of pistachios that were ozone-treated for 5 hours. According to results, 3 hours of ozonation and stirring of pistachios every 20 minutes is recommended as the best treatment for reducing microbial load, aflatoxin content, peroxide value and maintaining the sensory properties of dried pistachios.

Comparison of Different High-Speed Countercurrent Chromatography Injection Modes for the Separation of Glabridin.

In this work, different injection modes (single injection, overlapping injection, and continuous injection) of countercurrent chromatography were used and compared for the isolation and purification of glabridin from the crude extract of Glycyrrhiza glabra. The two-phase solvent system consisting of n-hexane/ethyl acetate/methanol/water (5:4:5:4, v/v) was employed as stationary and mobile phases. Compared with single injection mode, both overlapping injection and continuous injection modes exhibited higher separation efficiency at the same sample loading capacity. Their total separation times were 89.68% and 53.97% of the single injection mode, and the total solvent consumption were 74.88% and 52.49% of the single injection mode, respectively. Meanwhile, the yield, purity, and recovery rate of these three injection modes were almost the same. It was demonstrated that high-speed countercurrent chromatography in continuous injection mode was the most effective way for separating glabridin in three methods and it can be further applied to the separation of other economically valuable natural products.

Sample loading in gel electrophoresis using adapted 3D printers

In gel electrophoresis, samples that are dispensed too high above or too low into the wells result in sub-optimal outcomes. Here, an adapted 3D printer liquid handler equipped with an optical sensor was found to attain vertical sample delivery positionings at a standard deviation over mean ratio of 0.008. This illustrated high accuracy and repeatability outcomes achieved using automation that is cost effective and low in technical knowledge demand.

Impact energy absorption in 3D printed bio-inspired PLA structures

3D printing is an effective technique for designing and producing products with unique shapes that cannot be made using traditional methods. Polylactic acid (PLA) filaments, commonly used in 3D printing, are a promised material for fabricating composites and lightweight structures. This study investigates the fracture and damping properties of biodegradable PLA samples with a bio-inspired structure and different density under dynamical (ballistic) loading. PLA samples were fabricated using a 3D printer and a Fusion Deposition Modeling (FDM) method. A unique computer program for cellular samples with Schwartz-Diamond minimal surface topology was developed. The impact absorption energy under ballistic loading of PLA samples with Schwarz-Diamond surface structure was found to depend on their density and impact velocity. The internal structure of 3D-printed PLA cellular samples with a Schwarz-Diamond triply periodic minimal surface and the fracture mechanism of the cellular samples with different densities were demonstrated using scanning electron microscopy. It was found that the fracture mechanism changed from ductile to quasi-brittle with increasing sample density. This work provides a foundation for understanding the fabrication of plastic cellular products using 3D printing.

Purification of plasmid DNA using a novel two stage chromatography process

The chromatography process of large-scale plasmid purification with high efficiency and low cost has always been a major challenge. We established a two-step plasmid chromatography purification process combining multimodal and thiophilic chromatography with an overall chromatography yield of nearly 70%. Capto Core 700, a multimodal core–shell particle, was firstly used to remove the impurities from the crude lysate. The effects of different experimental conditions on chromatography recovery and impurity removal were screened. Compared to conventional size exclusion chromatography, the sample load and flow rate of this step were enhanced by 40-fold and 5-fold, respectively, while maintaining a 90% yield. For the thiophilic chromatography (Capto PlasmidSelect), the method of Design of Experiments (DoEs) was used to study the influence of parameters on the results. The effects of ammonium sulfate concentration, sodium chloride concentration and flowrate in the elution phase were studied and optimized with a central composite design model consisting of 17 experiments. The versatility of this process was demonstrated by successfully purifying three different lentiviral packaging plasmids (pLP1, pLP2 and pLP/VSVG) and the target plasmid containing green fluorescent protein (GFP). Purified plasmids consistently achieved a supercoiled purity of at least 90% with endotoxin levels below 5 EU/mg. Lentiviral vectors packaged using these plasmids exhibited high infectious titers of 1 × 107 TU/mL, thereby verifying the process applicability for diverse plasmid purification requirements.

Impact response of pseudoelastic nitinol

The response of polycrystalline nitinol with solely austenite structure was studied in three series of planar impact tests characterized by loading of the nitinol samples of 0.5–10 mm thickness by 1 mm thick aluminum impactor accelerated up to velocities of about 387, 429, and 567 m/s. In all the tests, the velocities of the free surfaces of the samples were monitored by a laser velocity interferometer. It was found that in all three test series, the amplitude of elastic precursor wave, being initially greater than 4 GPa, rapidly decays with the propagation distance down to ∼2.5 GPa, below which the decay is hindered by atomic clusters of the nanometer size. Based on the part of the velocity histories indicating the shock-induced austenite–martensite transformation, the initial, of about 2.5 × 103 s−1, and the maximum, up to 1 × 105 s−1, rates of the transformation were determined. As well, the impact stress slightly greater than 4 GPa was determined as that required for the onset of the B2 → B19′ transformation under shock loading. The unloading parts of the same velocity histories allowed a rough estimate of the fraction of the shock-transformed martensite and the elucidation of the virtually complete reversibility of the transformation.

Towards routine proteome profiling of FFPE tissue: insights from a 1,220-case pan-cancer study.

Proteome profiling of formalin-fixed paraffin-embedded (FFPE) specimens has gained traction for the analysis of cancer tissue for the discovery of molecular biomarkers. However, reports so far focused on single cancer entities, comprised relatively few cases and did not assess the long-term performance of experimental workflows. In this study, we analyze 1220 tumors from six cancer entities processed over the course of three years. Key findings include the need for a new normalization method ensuring equal and reproducible sample loading for LC-MS/MS analysis across cohorts, showing that tumors can, on average, be profiled to a depth of >4000 proteins and discovering that current software fails to process such large ion mobility-based online fractionated datasets. We report the first comprehensive pan-cancer proteome expression resource for FFPE material comprising 11,000 proteins which is of immediate utility to the scientific community, and can be explored via a web resource. It enables a range of analyses including quantitative comparisons of proteins between patientsand cohorts, the discovery of protein fingerprints representing the tissue of origin or proteins enriched in certain cancer entities.

Role of gut microbiome in developing necrotizing enterocolitis.

Necrotizing enterocolitis (NEC) is one of the most devastating intestinal diseases observed in preterm in the first days of life. Researchers have recently focused on potential predictive biomarkers for early and concomitant diagnoses. Thus, we inquired about the linkage of intestinal dysbiosis, one of the most important factors in NEC development to the gut microbiota. In this study, the systematic differences in the bacterial composition between neonates affected by NEC and healthy newborns were highlighted by metagenomic analysis. The next-generation sequencing of the V3-V4 variable region of the 16S rRNA gene and gene-specific qPCR analyzed the untargeted gut microbiota. Total bacteria, total and fecal coliform loads in stool samples with NEC were higher than control. OTU-level relative abundances of NEC infant was characterized by Firmicutes and Bacteroidetes at phylum levels. At the genus level, NEC stool was identified by the lack of Klebsiella and the presence of Roseburia, Blautia, and Parasutterella. Finally, Clostridium fessum was the predominant species of Clostridium genus in disease and healthy specimens at the species level, whereas Clostridium jeddahitimonense was at NEC diagnosis. Despite a strong relationship between pathophysiology and characterization of gut microbiota at a clinical diagnosis of NEC, our results emphasize the broad difficulty in identifying potential biomarkers.

Major alteration of lung microbiome and the host responses in critically ill COVID-19 patients with high viral load

Patients with COVID-19 under invasive mechanical ventilation are at higher risk of developing ventilator-associated pneumonia (VAP), associated with increased healthcare costs, and unfavorable prognosis. The underlying mechanisms of this phenomenon have not been thoroughly dissected. Therefore, this study attempted to bridge this gap by performing a lung microbiota analysis and evaluating the host immune responses that could drive the development of VAP. In this prospective cohort study, mechanically ventilated patients with confirmed SARS-CoV-2 infection were enrolled. Nasal swabs (NS), endotracheal aspirates (ETA), and blood samples were collected initially within 12 h of intubation and again at 72 h post-intubation. Plasma samples underwent cytokine and metabolomic analyses, while NS and ETA samples were sequenced for lung microbiome examination. The cohort was categorized based on the development of VAP. Data analysis was conducted using RStudio version 4.3.1. In a study of 36 COVID-19 patients on mechanical ventilation, significant differences were found in the nasal and pulmonary microbiome, notably in Staphylococcus and Enterobacteriaceae, linked to VAP. Patients with VAP showed a higher SARS-CoV-2 viral load in respiratory samples, elevated neutralizing antibodies, and reduced inflammatory cytokines, including IFN-δ, IL-1β, IL-12p70, IL-18, IL-6, TNF-α, and CCL4. Metabolomic analysis revealed changes in 22 metabolites in non-VAP patients and 27 in VAP patients, highlighting D-Maltose-Lactose, Histidinyl-Glycine, and various phosphatidylcholines, indicating a metabolic predisposition to VAP. This study reveals a critical link between respiratory microbiome alterations and ventilator-associated pneumonia in COVID-19 patients with higher SARS-CoV-2 viral loads in respiratory samples, elevated neutralizing antibodies, and reduced inflammatory cytokines, including IFN-δ, IL-1β, IL-12p70, IL-18, IL-6, TNF-α, and CCL4. These findings provide novel insights into the underlying mechanisms of VAP, with potential implications for management and prevention.

Design, development, and validation of a strand-specific RT-qPCR assay to differentiate replicating versus nonreplicating Rabies virus

The Rabies virus, a single-strand RNA virus with a negative-sense polarity, is responsible for causing encephalitis and is a zoonotic disease. If not promptly treated after infection, it has a close to 100 % fatality rate. Similar to other negative-sense polarity single-strand RNA viruses, the Rabies virus requires the creation of a positive-strand RNA intermediate for replication. One approach to identify this replication activity is to detect the complementary strand of the viral RNA genome in suspected infected cells or tissues. The reported Rabies virus RT-qPCR detection methods are designed to detect total viral load in samples without distinguishing between positive- and negative-strand for RNA viruses. As such, in this study, a sensitive Taqman-based strand-specific RT-qPCR assay has been developed to quantitatively detect both the positive- and negative-strand of the Rabies virus. This method demonstrates good reproducibility across a wide dynamic range and exhibits linearity of 8 logs with a lower limit of detection of 103 copies/μL for the positive-strand and 9 logs with a lower limit of detection of 102 copies/μL for the negative-strand. Notably, it can accurately detect a specific viral RNA strand even in the presence of high levels of the opposite strand, confirming the method’s specificity. In summary, a reliable strand-specific RT-qPCR assay has been developed and validated to differentiate replicating from non-replicating Rabies virus.

Optimizing reverse water gas shift catalysis: Minimizing metal loading and enhancing performance with Pt/ZrO2 catalysts through Rh incorporation

In this study, the optimization of catalysts for the reverse water gas shift reaction by minimizing metal loading and enhancing the performance of supported Pt catalysts through Rh incorporation was done. The catalysts were synthesized by the wet impregnation method, in which the metal noble load in bimetallic samples decreased by approximately one-third compared to the reference platinum catalyst, with varying Rh-Pt ratios. In all materials, m-ZrO2, obtained by the hydrothermal method, was used as support. In the catalytic tests in a continuous packed-bed flow reactor at atmospheric pressure, all samples were active to catalyze CO2 reduction between 200 and 300 °C. Bimetallic samples, particularly those with an Rh-Pt atomic ratio of 20–80, exhibited superior performance in the RWGS reaction despite their lower metal content. Based on the results of H2-TPR, FTIR in situ, and XPS, we propose that the enhanced CO production in bimetallic samples compared to Pt/ZrO2 catalysts can be attributed to Rh increasing the reduction extent of supported Pt. This enhancement facilitates the hydrogenation of bicarbonate species into formate species, which subsequently evolve into Pt0-CO and ultimately into CO. Additionally, Rh incorporation enables an alternative pathway for CO generation, where Pt0-carbonyls, known for their CO-producing capability, are formed via CO2 dissociation on the Pt0 surface. However, higher Rh concentrations favor the CO2 methanation rather than the RWGS reaction.

Challenges of Congenital HHV6 Infection Diagnosis and Treatment: Two Case Reports and Literature Review

Introduction: Congenital human herpesvirus 6 (HHV6) infection occurs in 1% of the general population and may result from the transmission of an inherited chromosomally integrated HHV6 (iciHHV6) or transplacental infection. It is mostly asymptomatic. Case reports: Case 1: a 29th-week-old female preterm newborn, admitted to the neonatal intensive care unit, became clinically unstable and irritable on the 20th day of hospitalization. Cranial ultrasound, revealed a significant posthemorrhagic tetraventricular dilation, with signs of ventriculitis. Investigations revealed HHV6 positivity on cerebrospinal fluid polymerase chain reaction multiplex panel testing and HHV6-DNA high viral loads in plasma samples. Case 2: a female late preterm newborn was admitted to the neonatal intensive care unit due to early-onset sepsis. Investigations revealed group B streptococcus positive blood cultures and cerebrospinal fluid HHV6 positivity on polymerase chain reaction multiplex panel testing, with negative bacterial culture. After 3 days of adequate antibiotic treatment, she maintained persistent moaning, which motivated a cranial ultrasound, revealing mild brain edema. Clinical improvement was observed only after beginning antiviral treatment in both newborns. Due to the persistency of high viral loads in both cases, despite antiviral treatment and clinical improvement, an iciHHV6 was suspected and posteriorly confirmed. Discussion/conclusion: Congenital iciHHV6 infection diagnosis is challenging because the presence of an iciHHV6 results in persistently high viral loads, even in the absence of active infection. Only a few diagnostic techniques can confirm active replication; unfortunately, these are not available in most countries. The decision to initiate antiviral treatment should be based on clinical judgment. Better ways for the diagnosis of active infection are needed.

Aluminium oxide/aluminium cobalt oxide as an efficient electrocatalyst for the electrochemical detection of 4-nitroaniline in environmental samples

The rise of pollutants in our ecosystem has become a major threat to be resolved. 4-nitroaniline (4-NAE) is among the carcinogenic and toxic components present in the environment with harmful effects. Thus, we designed an electrochemical sensor to detect 4-NAE with the as-synthesized aluminium oxide/aluminium cobalt oxide (AO/ACO) via the hydrothermal preparation approach. Ensured with the physicochemical characterization of the sample (AO/ACO) with XRD, FTIR, Raman, and FESEM, the presence of AO/ACO was confirmed with no impurities. The electrochemical analysis of 4-NAE was analyzed after the optimization of the sample loading. The CV and DPV results signify that the developed sensor has excellent repeatability and reproducibility for 4-NAE detection with a low detection limit of 0.007 µM and a sensitivity of about 17.96 µAµM−1cm−2. The selectivity test studied with the presence of various interfering compounds showed satisfactory results with no interference other than 4-NAE. The environmental samples applied for the spiked 4-NAE showed good results proving the sensor's ability to detect 4-NAE in real samples. Thus, the sensor will be a reliable technique for the implementation in environmental applications for the detection of 4-NAE and even other compounds with modifications.

Increasing accuracy in predicting mode I fracture toughness of rock structures: a comparative analysis of the rock engineering system method

The investigation of crack initiation and expansion is vital for the stability of structures. The Mode I fracture toughness (KIc) of rocks is a key property used to predict crack propagation in tension and hydraulic fracturing. Various methods have been introduced to determine KIc, but results differ due to factors like sample dimensions, crack geometry, groove type, and loading conditions. The cracked chevron notched Brazilian disc (CCNBD) sample is commonly used in laboratory tests for its easy preparation. This study employs the rock engineering system (RES) technique to overcome the challenges of time-consuming and costly laboratory tests and the uncertainty in traditional methods (analytical, numerical, experimental, laboratory, regression). Using 88 CCNBD rock samples proposed by ISRM, input parameters include thickness of the disc specimen (B), uniaxial tensile strength (σt), initial crack length (α0), radius of the disc specimen (R), crack length (αB), and the length of the final crack (α1). The RES-based model used 70 data points (80% of the dataset) for development, and 18 data points (20%) for evaluation. Regression analysis compared the performance of the RES method, using statistical indicators such as squared correlation coefficient (R2), mean square error (MSE), and root mean square error (RMSE) to measure accuracy. The RES-based method outperformed other regression techniques, demonstrating significantly enhanced accuracy. This highlights the effectiveness and superior performance of the RES method in estimating fracture toughness, particularly for CCNBD samples, showcasing its potential as a robust analytical tool.

Physio-chemical and microbial assessment of the source of drinking water and the prevailing waterborne diseases in Gwagwalada Municipal Area Council of Abuja, Nigeria

Water contamination poses a serious threat to the wellbeing of people globally. In Nigeria, there have been increasing cases of waterborne diseases owing to water contamination, thus becoming public health concerns. This study investigated the physio-chemical properties, microbial loads of borehole water samples from Gwagwalada area council of Abuja and its prevailing waterborne diseases. Borehole water samples were collected from five wards of the council area (Gwagwalada central (YZTM), University of Abuja Staff quarters (YZTMB), Dobi (SBWTM), Tunga Maje (TMCBW), and Ikwa (YZBTM) in triplicate and analyzed for physiochemical properties (temperature, conductivity, pH, turbidity, salinity, total dissolved solids (TDS), and heavy metals) and microbial contamination using standard methods. Surveillance to determine the prevalence of waterborne diseases was also carried out in the health facilities of the area council. All samples showed variable concentrations in most of the physiochemical parameters and were within the recommended thresholds except for conductivity (596.01 ± 0.00 μS/cm) and chlorine (254.18 ± 0.00 mg/l) in TMCBW sample and total dissolved solids in TMCBW (948.01 ± 14.14 mg/l) and YZTM (GC) (1871.00 ± 55.15 mg/l) water samples. E. coli, Enterobacter spp., Shigella spp., Lactobacillus spp., Pseudomonas ssp., and Proteus ssp. were bacteria isolates of the samples. Principal component analysis of interrelationship showed a strong positive relationship between some physio-chemical parameters and sampling locations. Also, there was heavy burden of gastrointestinal (waterborne) diseases. The study concludes that the borehole samples from the area were contaminated and maybe unfit for consumption. Thus, call for interventions that provide safer portable water or treat existing water sources towards achieving SDG No. 6 and to prevent outbreaks of waterborne diseases that could emanate from the consumption of unsafe water.

Evaluation of in-house-built pipette-tip micro-solid-phase extraction devices for sample preparation in the analysis of amino compounds by liquid chromatography-tandem mass spectrometry

Amino compounds are of significant interest in dietary, clinical, and quality control fields. Efficient extraction is crucial for comprehensive metabolomics, especially for amino acids and biogenic amines, but traditional solid-phase extraction (SPE) methods are costly and require large solvent volumes. Miniaturized SPE techniques, like pipette-tip micro-solid-phase extraction (PT-µ-SPE), offer promising alternatives by improving throughput and reducing solvent and sorbent usage. This study presents PT-µ-SPE for the screening and quantification of amino compounds in bee products, particularly honey. The method involves derivatization with diethyl ethoxymethylenemalonate (DEEMM) and analysis using liquid chromatography-triple quadrupole mass spectrometry. Silica-based SCX sorbents were effective for a broad range of amino compounds, while WCX sorbents were better for aliphatic amines. The method's extraction efficiency was assessed across sample loading, washing, and elution solution, with recovery rates of 70 - 120% in oat bran, sea buckthorn leaves, and honey extracts. Matrix effects were observed for four amino compounds in honey. Limits of detection (LoD) and quantification (LoQ) ranged from 0.37 to 57 µg L⁻¹ and 1.13 to 174 µg L⁻¹, respectively. Covering 48 amino compounds under different PT-µ-SPE conditions, this method has been applied to several samples, demonstrating accuracy, environmental sustainability, cost-effectiveness, portability, and versatility in amino compound analysis.

Volumetric printing and non-destructive drug quantification of water-soluble supramolecular hydrogels.

Vat photopolymerisation 3D printing is being actively explored for manufacturing personalised medicines due to its high dimensional accuracy and lack of heat application. However, several challenges have hindered its clinical translation, including the inadequate printing speeds, the lack of resins that give soluble matrices, and the need for non-destructive quality control measures. In this study, for the first time, a rapid approach to producing water-soluble vat photopolymerised matrices and a means of non-destructively verifying their drug content were investigated. Volumetric printing, a novel form of vat photopolymerisation, was used to fabricate personalised warfarin-loaded 3D-printed tablets (printlets). Eight different formulations containing varying amounts of warfarin (0.5-6.0% w/w) were used to print two different sized torus-shaped printlets within 6.5 to 11.1s. Nuclear magnetic resonance (NMR) spectroscopy revealed the presence of only trace amounts of unreacted acrylate monomers, suggesting that the photopolymerisation reaction had occurred to near completion. All printlets completely solubilised and released their entire drug load within 2.5 to 7h. NIR spectroscopy (NIRS) was used to non-destructively verify the dose of warfarin loaded into the vat photopolymerised printlets. The partial least square regression model built showed strong linearity (R2 = 0.980), and high accuracy in predicting the drug loading of the test sample (RMSEP = 0.205%). Therefore, this study advances pharmaceutical vat photopolymerisation by demonstrating the feasibility of producing water-soluble printlets via volumetric printing and quantifying the drug load of vat photopolymerised printlets with NIRS.

Microcolumn and coelution hydration of oil seal blood spot for efficient screening of newborn α-thalassemia via chip isoelectric focusing

BackgroundThe global prevalence of α-thalassemia necessitates effective newborn screening strategies due to its severe clinical consequences. Traditional methods such as liquid chromatography (LC), capillary electrophoresis (CE), and isoelectric focusing (IEF) face limitations, including low separation efficiency, poor sensitivity for detecting Hb Bart's, and time-intensive operations, particularly with dried blood spots (DBS). These limitations hinder timely and accurate screening. This study addresses the need for a more efficient, sensitive, and rapid method for detecting Hb Bart's in newborns. ResultsWe enhanced IEF separation and sensitivity by designing a microfluidic IEF (mIEF) system with shortened columns and employing a coelution sample loading technique using oil-sealed blood spots for rapid sample pretreatment. Our experiments demonstrated significant improvements: the total analysis time was reduced from 1110 min (IPG IEF) and 46 min (LC) per batch to 36 min per batch. For individual samples, the focusing time decreased from 6 min (previous mIEF) to 3 min, with the microcolumn length shortened by 50 %, from 30 mm to 15 mm. The developed method showed excellent consistency with clinical Bart's detection and PCR diagnosis, achieving 100 % sensitivity and 98 % specificity for α-thalassemia screening. Significance and noveltyOur novel mIEF method provides an efficient, sensitive, and rapid tool for screening newborns for α-thalassemia. This advancement addresses the limitations of traditional techniques, improving early diagnosis and intervention strategies and ultimately enhancing health outcomes for at-risk newborns.