Sample Volume Research Articles

Detection of renal tubular transporter proteins in canine urinary extracellular vesicles using liquid chromatography-tandem mass spectrometry.

Urinary extracellular vesicles (UEVs) are membranous particles that carry renal tubular transporter proteins. Here, we evaluate whether selected renal tubular transporter proteins can be detected in UEVs isolated from small volume (1-5 mL) canine urine samples of healthy dogs and canine patients with elevated circulating parathyroid hormone (PTH)/PTH-related peptide (PTHrp) concentrations, hypercortisolism, and primary hypoadrenocorticism using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The total creatinine content of each urine sample was calculated from urine volume and creatinine concentration. UEVs were isolated by size exclusion chromatography prior to quantification by nanoparticle tracking analysis and proteomic analysis by LC-MS/MS. Group comparisons were made using non-parametric statistics. Aquaporin-2 (AQP2) and the renal sodium/phosphate co-transporter (NPT2A) were detected in UEVs isolated from small volume samples of almost all healthy dogs but were not detected in most dogs with elevated circulating PTH/PTH related peptide (PTHrp) concentrations, hypercortisolism and primary hypoadrenocorticism. Total creatinine content of the urine sample was strongly positively correlated with the number of UEVs (rs = .84, P < .001); thus, total creatinine was used as a surrogate marker of UEV number. In healthy dogs, AQP2 and NPT2A were both detected in samples containing at least 1.7 × 109 UEVs or 24 μmol creatinine, however in non-healthy dogs, AQP2 and NPT2A were not detected in most samples containing up to 6.3 × 109 UEVs or 32 μmol creatinine.

Rapid and eco-friendly microextraction procedure based on green hydrophobic deep eutectic solvent for simultaneous determination of five sex hormones from cosmetic samples

Hormones, which are among the endocrine disruptors, are used in many fields and their use in cosmetic products has recently attracted considerable attention. Despite the negative health consequences, there is no detailed guidance that limits the use of these endocrine disruptors. This study aimed to develop a green hydrophobic deep eutectic solvent (HDES)-based microextraction method for the detection and quantification of sex hormones (progesterone, β-estradiol, estrone, ethinylestradiol, and testosterone) in different cosmetic products. The best value for extraction recovery was obtained by using DL-menthol and dodecanoic acid as green HDES in a 4:1 mole ratio. Optimization parameters such as HDES volume, pH, sample volume, centrifugation time, mixing type, and mixing time were investigated for cosmetic samples to be used as real samples. Furthermore, the extraction recovery values ranged from 85.7 % to 101.3 % and the detection limit was between 2.33–3.30 ng mL−1. Additionally, three different greenness methods, namely ComplexGAPI, AGREEprep, and BAGI, were used to evaluate how green the proposed method is in this study. The microextraction procedure developed in this study was successfully applied to five different cosmetic samples that will be used as real samples to detect and determine sex hormones.

Development and Validation of an Improved HPLC-MS/MS Method for Quantifying Total and Unbound Lenalidomide in Human Plasma.

This study aimed to develop a fully validated HPLC-MS/MS method for quantifying total and unbound lenalidomide concentrations in human plasma. Unbound concentrations were measured using plasma ultrafiltrate prepared with Amicon® Centrifugal Filters. Lenalidomide and lenalidomide-d5 (internal standard) were extracted from 50 μL of human plasma using liquid-liquid extraction. Chromatography was conducted with a Halo® C18 column using 0.1% formic acid and methanol (20:80, v/v) as the mobile phase. The mass spectrometer was operated in a positive ion mode with an electrospray ionization interface and multiple reaction monitoring modes. Calibration curves were linear over the range of 5 to 1000 ng/mL (r2 > 0.996) for both the total and unbound lenalidomide. For total lenalidomide concentrations, between-run precision (coefficients of variation) and accuracy were 1.70-7.65% and 94.45-101.10%, respectively. For unbound concentrations, inter-day precision and accuracy were 1.98-10.55% and 93.95-98.48%, respectively. We developed a highly reproducible, sensitive, and efficient bioanalytical method using a smaller volume of plasma sample (50 μL) with a relatively short run time (2.5 min). The proposed analytical method was successfully applied to measure total and unbound lenalidomide concentrations at various time points in multiple myeloma patients with renal impairment.

Novel nanoliter spray enhanced microwave plasma ionization mass spectrometry for the simultaneous detection of heavy metals and organic plasticizers in soil: A case study in a lead-acid battery industrial park

Soil pollution is predominantly attributed to the presence of heavy metal elements and organic compounds; However, current detection methodologies are restricted to the identification of only one of these two sources at a time. A novel analytical approach, known as nanoliter spray enhanced microwave plasma ionization mass spectrometry (Nano-Spray-EMPI-MS), has been developed to facilitate the simultaneous detection of both heavy metals and organic pollutants in soil samples. This technique is characterized by its requirement for minimal sample volumes, thereby allowing for efficient and rapid analysis. The research concentrated on the simultaneous analysis of five heavy metals (Pb, Zn, Cu, Cr, and Ni) and three major phthalates (PAEs), specifically DEHP, DBP, and DMP. The detection and quantification limits for the heavy metals were established to be between 0.16-0.57 and 0.53–1.88 μg L−1, respectively, while the limits for the PAEs ranged from 0.02 to 0.05 and 0.07–0.16 μg L−1. Validation of the method's efficacy in soil detection demonstrated recovery rates of 90.9 %–105.7 % for heavy metals and 89.4 %–97.2 % for PAEs. The application of this method analyzing soil samples collected from an area adjacent to a lead-acid battery industrial park in China revealed varying levels of contamination by both heavy metals and PAEs. Notably, Lead contamination was found to be the most pronounced, with a peak concentration of 862.5 mg kg−1 and a correspondingly high pollution index. These findings are significant for evaluating local ecological risks, pinpointing sources of pollution, and formulating effective pollution management strategies in the region.

Determination of a plasma retinol reference interval in eastern collared lizards (Crotaphytus collaris)

BackgroundThe Eastern collared lizard (Crotaphytus collaris) is a species of lizard native to the United States and Mexico and is growing in popularity in the exotic community around the world. There is minimal knowledge known on the analyte breakdown of this or closely related species. As this species becomes more popular in the exotic community more medical conditions are being identified that can be assisted in diagnosis with the establishment of ranges of the different essential analytes. Being an insectivorous lizard, this species may suffer from hypovitaminosis A. There has been a lack of antemortem diagnostics available for minimally invasive and appropriate diagnosis of this disease. Recent studies have been performed in other small insectivore species establishing plasma retinol reference intervals. Reference intervals for retinol have not been published or validated in collared lizards or any closely related species. MethodsPlasma, with a sample volume of only 50 µl, from forty clinically healthy captive bred Eastern collared lizards, twenty males and twenty females were analyzed using ultra-high-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) to determine plasma retinol concentrations. ResultsUsing UPLC-MS/MS analysis, data was normally distributed and a reference interval for plasma retinol in the Eastern collared lizard was 1.57-3.07 µg/mL (0.237-0.464 µmol/L). A mean of 0.31µg/ml; with a lower limit 95% confidence interval being 1.34-1.80 µg/ml and upper limit of 2.84-3.30 µg/ml. Vitamin A (retinol) levels were significantly different when comparing a captive bred line called Blazing Blue (n=4), with all other sampled captive bred localities. Including a closely related Aquaflames (n=16), (Sky Blue) (n=2), Western yellowhead (n=5), Fancy Spotted (n=4), Aquamarine (n=3), and Lightning Yellow (n=6); with the Blazing Blue morph being significantly lower using a t-Test comparison assuming equal variations. There was only one statistical variation seen in age groups between the 1-year-old and 6-year-old age groups with the 6-year-old age group being significantly lower than the 1-year-old age group. There was no variation based on sex. Conclusions and clinical relevanceThis study provides the first established plasma retinol range in a species in the suborder iguania. Values are valuable in assistance in diagnosing and treating hypovitaminosis A and to provide advice on dietary supplementation and dietary improvement for the collared lizards and possibly other related species as well. In addition, these values can be used to design and evaluate other studies for nutritional needs of this species. Limitations of this study include only a single season of collection, no same time comparison to liver values, some age groups were limited to only one individual, and the evaluation of only captive bred individuals.

Culture failure and suboptimal analysis in cytogenetics: A data review as a quality improvement metric from a resource-limited country

Abstract Introduction/Objective Conventional cytogenetic failures can affect hematological cancer diagnosis and prognosis. Resource-constrained regions without contemporary genetic testing facilities may experience this impact more. The effectiveness of this technique is contingent on cultural success, availability of metaphases, and morphological adequacy. The results can be influenced by the underlying diagnosis, inherent biological features of the leukemic clone, sample conditions, and transit time. Culture failure (CF) and suboptimal analysis (SOA) data can identify problems and set standards. Thus, we designed a cohort to find areas of improvement and reduce negative contributing factors, making this cytogenetic technique more accessible and cost-effective. Methods/Case Report A retrospective study was conducted at Cytogenetics laboratory at Indus hospital, Karachi. From January 2021 to March 2023, 1234 blood and bone marrow samples were processed. CF was defined as no growth, whereas suboptimal cases showed a low mitotic index or poor morphology. Data analysis included collection and processing time, sample volume, cell count, specimen type, in-house or out-reach sample, and diagnosis. Armonk, NY-based IBM SPSS 24.0 used to analyze the data. Chi-square used to find association of various factors with karyotyping. A p-value &amp;lt; 0.05 indicated significance. Results (if a Case Study enter NA) Of 1234 samples, 1110 (90%) were bone marrow and 124 (10%) were peripheral blood. There were 32/1234 (2.6%) CF cases. Each CF case was found in bone marrow samples, making the true incidence 32/1110 (2.9%). Constitutional blood tests found no CF in 124 samples. Additionally, chromosomal analysis quality was assessed. SOA occurred in 105/1234 (8.5%) instances, with 58% having poor morphology and 31% having a low mitotic index. Among 137 CF and SOA patients, 134/1110 (12%) were found in marrow and 3/124 (2.4%) in blood. Conclusion Lower CF rates suggest that appropriate standards and quality management may reduce cytogenetic failure rates despite resource constraints and newly established services.

Microfluidic-Integrated Chip Resonators for Electron Spin Sensing in Submicromolar, Submicroliter Solutions.

Planar or chip microresonators decrease the sample volume required for magnetic resonance spectroscopies to the nanoliter scale. However, the interrogation of nanoliter-scale solution samples on planar sensors is hindered by the lack of microfluidic devices that can simultaneously provide a small total volume and long-term sample stability. Here, we report microfluidic devices that decrease the total required sample volume to the submicroliter scale and also provide long-term physical stability and storability. We also report a 3D-printed microfluidic with a self-contained actuation mechanism, which allows the sample to be retracted from the microresonator surface for storage. The microfluidic devices are fabricated easily by laser cutting or 3D printing and are integrable with a broad range of planar sensors. We use planar inverse anapole (PIA) microresonators to obtain continuous wave (CW) electron paramagnetic resonance (EPR) spectra of natural-isotopic-abundance nitroxide radicals, which are ubiquitously used as reporters of biomolecular dynamics. We provide experimental evidence for a concentration sensitivity of 330 ± 40 nmol L-1, a concentration sensitivity limit of 800 ± 100 nmol L-1/mT√Hz, and an active volume no greater than 30 nL. Together, these developments represent an advance not only in the sensitivity of EPR spectroscopy but also in the design of microfluidics for stable, dead-volume-free placement of nanoliter-scale volumes of solutions on planar sensors.

In-Depth Chemical Analysis of the Brain Extracellular Space Using In Vivo Microdialysis with Liquid Chromatography-Tandem Mass Spectrometry.

Metabolomic analysis of samples acquired in vivo from the brain extracellular space by microdialysis sampling can provide insights into chemical underpinnings of a given brain state and how it changes over time. Small sample volumes and low physiological concentrations have limited the identification of compounds from this compartment, so at present, we have scant knowledge of its composition. As a result, most in vivo measurements have limited depth of analysis. Here, we describe an approach to (1) identify hundreds of compounds in brain dialysate and (2) routinely detect many of these compounds in 5 μL microdialysis samples to enable deep monitoring of brain chemistry in time-resolved studies. Dialysate samples collected over 12 h were concentrated 10-fold and then analyzed using liquid chromatography with iterative tandem mass spectrometry (LC-MS/MS). Using this approach on dialysate from the rat striatum with both reversed-phase and hydrophilic interaction liquid chromatography yielded 479 unique compound identifications. 60% of the identified compounds could be detected in 5 μL of dialysate without further concentration using a single 20 min LC-MS analysis, showing that once identified, most compounds can be detected using small sample volumes and shorter analysis times compatible with routine in vivo monitoring. To detect more neurochemicals, LC-MS analysis of dialysate derivatized with light and isotopically labeled benzoyl chloride was employed. 872 nondegenerate benzoylated features were detected with this approach, including most small-molecule neurotransmitters and several metabolites involved in dopamine metabolism. This strategy allows deeper annotation of the brain extracellular space than previously possible and provides a launching point for defining the chemistry underlying brain states.

Are you filtering enough? Unveiling the impact of water sample volumes on eukaryotic microbial community in freshwater lakes

The appropriate volume of filtered water samples is crucial for accurately reflecting microbial community characteristics. Previous research has shown that changes in water sample volumes significantly affect bacterial diversity and the relative abundance of dominant taxa in marine environments. However, the specific impact on freshwater microbial diversity and community structure, particularly for eukaryotic microorganisms, remains unclear. To address this gap, we collected water samples from eutrophic Lake Taihu and mesotrophic Lake Bosten. We filtered the samples through 0.2 μm filters using volumes ranging from 0.1 to 2 L for Lake Taihu and 0.1 to 3.2 L for Lake Bosten. Analysis of 18S rRNA gene amplicons revealed that variations in filtered water sample volume significantly affected the diversity, community structure, and composition of eukaryotic microorganisms. In light of these findings, we recommend using a filtered water sample volume of approximately 0.2 L for eutrophic aquatic ecosystems and 0.8 L for mesotrophic aquatic ecosystems in microbial analyses. These recommendations underscore the importance of selecting the appropriate filtered water sample volume to ensure representative and reliable results in the study of eukaryotic microorganisms in freshwater ecosystems.

Synergy of high-intensity chromium ion implantation and ion beam energy impact on the zirconium alloy surface

The peculiarities and modes of material modification with high-intensity, high-power density ion beams on the irradiated surface are studied for the first time. Chromium ions are implanted into a zirconium alloy using a 25 kW/cm2, 450 μs beam at the pulse repetition rates within 8–35 pps. Every high-energy ion pulse impact is followed by ultrafast cooling of the surface due to heat removal into the target material. Three modes are studied at the temperatures of 580, 700, and 900 °C with an additional pulsed heating. An increase in the average target temperature from 580 to 700 °C within 1 h at the same pulse power density allows increasing the depth of chromium ion alloying from 1.5 to more than 7 μm. The use of ultrafast cooling of the Zr1%Nb alloy surface offers a grain size reduction from a few μm to approximately 50–250 nm, without any microstructural changes throughout the sample volume. An inhomogeneous chromium ion distribution over the target surface and depth is observed.

Investigation of geochemically induced permeability alteration in geothermal reservoirs and its implications for sustainable geothermal energy production

Geothermal resources constitute a significant portion of the world's low-carbon, renewable energy potential, with about 75% classified as low-temperature. One such potential resource exists in Precambrian basement rocks underlying the Williston Basin in southern Saskatchewan, Canada, with a reservoir temperature of 120 °C. However, geochemically induced permeability alteration in these highly reactive low-temperature granitoid resources poses a significant risk to long-term heat production. To assess and potentially mitigate this risk, we conducted a geochemical and mineralogical study of both altered and unaltered samples. Our findings facilitated the parameterization of geochemical simulations of water-rock interactions to predict mineral volume changes and, by extension, draw inferences on porosity and permeability changes resulting from these interactions. The simulations indicate an increased mineral volume in both samples, yet geothermal alteration of the unaltered, and thus more reactive, rocks induced relative mineral volume changes about 30% greater than those in the altered rocks. The resulting absolute change in porosity is 0.5 vol% for the unaltered rocks and 0.35 vol% for the altered rocks. Utilizing an empirical porosity-permeability relationship, the computed change in permeability indicates that the unaltered basement rock experienced a greater change in total permeability than the altered basement rocks. Additional calculations demonstrate the sensitivity of the porosity-permeability equation to critical porosity and power exponent, forecasting various scenarios with permeability changes ranging from 1.0 × 10−13 to 1 × 10−20 m2. Consequently, we infer that altered, permeable zones of the examined Precambrian basement rocks are likely to offer favourable conditions for sustained, multi-decade heat production, and thus should be targeted over less altered zones to justify initial capital expenditures. Globally, geothermal heat extraction from these rocks remains an underexplored yet promising resource for generating reliable, low-carbon renewable energy, crucial in our efforts to decarbonize the global economy.

Preparation and Application of Polyclonal Antibodies for the Rapid Detection of Actinidia Chlorotic Ringspot-Associated Virus

Actinidia chlorotic ringspot-associated virus (AcCRaV, Emaravirus actinidiae) is prevalent in Chinese kiwifruit, leading to substantial yield reduction. The intricate nature of symptoms presents diagnostic challenges, underscoring the necessity for a rapid and accurate detection method that facilitates effective control. In this investigation, AcCRaV isolates from key kiwi-producing regions in Sichuan province were collected and analyzed, with representative strains chosen as experimental materials. Primers targeting the nucleoprotein gene of AcCRaV were designed, and their codon usage was optimized to enhance performance. Various serological methods utilizing polyclonal antibodies were developed, including ELISA, dot immunobinding assay, and AcCRaV-specific gold immunochromatographic bands (AcCRaV-GICS). Field samples exhibited high specificity and sensitivity when tested using these methods. Furthermore, the results obtained from a large number of field samples are consistent with those derived from RT-PCR analysis, further validating the applicability of our approach. A detection method capable of handling a large volume of field samples infected with AcCRaV is currently lacking; thus, our system construction provides an important reference for addressing this gap.

IT-services: Green Means Competitive?

Introduction. The current stage of economic development is characterised by significant digitalisation and a noticeable shift of corporate strategies in favour of sustainable development. The first of these trends makes IT companies interesting for analysis, since in the era of digitalisation their impact on the global economy is significant and undeniable. The second trend is largely related to the exhaustion of tangible competitive factors and the shift of focus towards intangible competitive factors, in particular, towards catering to the specific needs of consumers, i.e. in the consideration of social and environmental factors throughout value chains. In line with modern trends, meeting these needs is achieved by implementing ESG-principles, including by IT companies, which is where the two major modern trends - digitalisation and corporate sustainability intertwine. In this context, the aim of the research is to analyse the relationship between IT companies’ financial performance and their ESG position.Materials and methods. The study is based on the analysis of correlation between the data of companies' financial statements and their valuations within the framework of S&amp;P ESG-rating. Results of the research. According to the results of the analysis on the basis of the presented data sample, no significant correlation between revenue and profit indicators (on the one hand) and ESG[1]assessments of S&amp;P rating (on the other hand) was revealed.Discussion and conclusion. The results obtained for companies in the IT-services industry generally correspond to the results of similar calculations by the authors for a number of other industries. Of course, they reflect the situation in a particular industry, with specific indicators (profit, revenue), taking into account such limitations of the study as the methodology of the selected ESG-rating and the volume of data sampling. Nevertheless, within the framework of the study we can say that there is no significant correlation between the financial performance of IT services companies and the implementation of ESG principles by these ccompanies.

Fabrication and Fluorescence Analysis of Rhodamine Dyes in Polycarbonate Serpentine Microfluidic System.

The synthesis of rhodamine dyes (R6G and R1010) and their fluorescence characterization within polymer-based microfluidics, offers an exciting and novel approach in materials science and chemical analysis. This work investigates the emission of polycarbonate substrates (PC) by UV-visible. The ablation threshold (16mj.sec-1) of PC at 193nm wavelength after that ablation process continued to produce microfluidic serpentine channels on PC by using G-Code. The fluorescence characteristics of Rhodamine 6G and Rhodamine 101 are investigated. Absorption and emission at peak wavelength were analyzed against R6G and R101 concentrations. Furthermore, the refractive indices of both R6G and R101 vis concentrations are examined. As a result at low concentrations, there was the highest overlapping, and at high concentrations, there was the smallest overlapping. R101 showed better photostability and a more consistent diffusion, whereas R6G had a faster diffusion and stronger fluorescence intensity. These differences were caused by the different molecular structures of the dyes and their interactions with the PCmicrochannel. Incorporating R6G and R101 dyes into a polycarbonate PC microfluidic chip would enhances both the resolution and sensitivity of fluorescence detection. The limited microfluidic setup facilitates ultra-high-resolution investigation and minimizing sample volumes, making it suitable for applications requiring precise measurements. The innovation relies on the utilization of the unique fluorescence characteristics of R6G (Rhodamine 6G) and R101 (Rhodamine 101) dyes to enhance the performance of polycarbonate microfluidic devices. R6G has high fluorescence quantum yield and stability, rendering it suitable for sensitive detection, while R101 offers superior brightness and improved resistance to photobleaching. Incorporation of these dyes into polymeric microfluidics improves sensitivity and facilitates real-time, dynamic sample analysis. This method offers a portable, economical solution with high-throughput capabilities, greatly enhancing both analytical and process accuracy across a variety of applications.

Enhanced Pb(II) extraction using a novel chitosan/pectin/carbon composite adsorbent for solid-phase extraction

ABSTRACT This research aimed to present the development of chitosan/pectin/carbon (CPC) materials as adsorbents in the solid-phase extraction (SPE) of Pb(II) from river water samples. The CPC adsorbent shows active groups effectively binding Pb(II) ions. The characterisation included using Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy with Energy-Dispersive X-ray Spectroscopy (SEM-EDX) to analyse functional groups and evaluate surface morphology with elemental composition. SPE parameters were systematically optimised to enhance adsorption-desorption efficiency, including pH 4.0, Pb(II) concentration of 5 mg L−1, flow rate of 13.3 mL min−1, elution flow rate of 12.8 mL/min using nitric acid (HNO3), a sample volume of 25 mL, eluent volume of 1 mL and concentration at 1.0 M. The results showed that the implementation of the refined CPC-based SPE method for Pb(II) analysis in water samples obtained a recovery range of 101% to 103%, accompanied by a significant 25-fold concentration enrichment of Pb(II). Moreover, the adsorption concentration increased from undetectable levels to 1.27–2.48 mg L−1, reporting the efficacy of the CPC-based SPE method for sensitive and accurate Pb(II) analysis in environmental samples.

Resolving Structures of Paramagnetic Systems in Chemistry and Materials Science by Solid-State NMR: The Revolving Power of Ultra-Fast MAS.

Ultra-fast magic-angle spinning (100+kHz) has revolutionized solid-state NMR of biomolecular systems but has so far failed to gain ground for the analysis of paramagnetic organic and inorganic powders, despite the potential rewards from substantially improved spectral resolution. The principal blockages are that the smaller fast-spinning rotors present significant barriers for sample preparation, particularly for air/moisture-sensitive systems, and are associated with low sensitivity from the reduced sample volumes. Here, we demonstrate that the sensitivity penalty is less severe than expected for highly paramagnetic solids and is more than offset by the associated improved resolution. While previous approaches employing slower MAS are often unsuccessful in providing sufficient resolution, we show that ultra-fast 100+kHz MAS allows site-specific assignments of all resonances from complex paramagnetic solids. Combined with more reliable rotor materials and handling methods, this opens the way to the routine characterization of geometry and electronic structures of functional paramagnetic systems in chemistry, including catalysts and battery materials. We benchmark this approach on a hygroscopic luminescent Tb3+ complex, an air-sensitive homogeneous high-spin Fe2+ catalyst, and a series of mixed Fe2+/Mn2+/Mg2+ olivine-type cathode materials.

Ultrasound Imaging of Nonlinear Media Response Using a Pressure-Dependent Nonlinearity Index

It has been shown that within the range of acoustic pressures used in ultrasound imaging, waveforms are distorted during propagation in tissue due to the physically nonlinear behavior of the tissue. This distortion leads to changes in the spectrum of the received ultrasound echoes, causing the transfer of signal energy from the fundamental frequency to higher harmonics. Interestingly, adipose tissue exhibits up to 50 % stronger nonlinear behavior compared to other soft tissues. The tissue nonlinearity parameter B/A is typically measured ex vivo using an ultrasound method in transmission mode, which requires extensive receiving systems. Currently, there is no improved ultrasound method for measuring the B/A nonlinearity parameter in vivo, which could be used in assessing the degree of fatty liver disease. We propose a new, simple approach to estimating nonlinear tissue properties. The proposed method involves transmitting ultrasound waves at significantly different acoustic pressures, recording echoes only in the fundamental frequency band at various depths, and introducing a nonlinearity index (NLI) based on specific echo amplitude ratios. The NLI at a given depth is calculated using the ratio of two dimensionless parameters. The first parameter is a predetermined constant obtained by dividing the total echo values from transmitting a signal at higher sound pressure by those from a signal at lower sound pressure, summed over a small tissue sample volume located near the transducer. The second parameter is calculated at a fixed distance from the transducer, determined by dividing the total echo values from transmitting a signal at higher sound pressure by those from a signal at lower pressure, summed over a small tissue volume of the tissue at that distance from the transducer. The reliability of the proposed measurements for assessing tissue nonlinearity has been substantiated through experimental confirmation of the existing correlations between the values of NLI and B/A in water, sunflower oil, and animal liver tissue samples with oil-enriched regions. The NLI was more than 15 % higher in sunflower oil than in water. The NLI in bovine liver sample below the area with injected oil (mimicking “steatosis”) was more than 35 % higher than in regions without oil. This method represents a promising modality for the nonlinear characterization of tissue regions in vivo, particularly for diagnosing fatty liver disease.

The Miniaturized Isolation of Neutrophil Granules (MING) method allowed a deep proteome mapping of human neutrophil granules.

Neutrophils are the innate immune system's first line of defense, and their storage organelles are essential to their function. The storage organelles are divided into three different granule types named azurophilic, specific, and gelatinase granules, besides a fourth component called secretory vesicles. The isolation of neutrophil's granules is challenging, and the existing procedures rely on large sample volumes, about 400 mL of peripheral blood, precluding the use of multiple biological and technical replicates. Therefore, the aim of this study was to develop a miniaturized isolation of neutrophil granules (MING) method, using biochemical assays, mass spectrometry-based proteomics and a machine learning approach to investigate the protein content of these organelles. Neutrophils were isolated from 40 mL of blood collected from three apparently healthy volunteers and disrupted using nitrogen cavitation; the organelles were fractionated with a discontinuous 3-layer Percoll density gradient. The isolation was proven successful and allowed for a reasonable separation of neutrophil's storage organelles using a gradient approximately 37 times smaller than the methods described in the literature. Moreover, mass spectrometry-based proteomics identified 368 proteins in at least 3 of the 5 analyzed samples, and using a machine learning strategy aligned with markers from the literature, the localization of 50 proteins was predicted with confidence. When using markers determined within our dataset by a clusterization tool, the localization of 348 proteins was confidently determined. Importantly, this study was the first to investigate the proteome of neutrophil granules using technical and biological replicates, creating a reliable database for further studies.

Exploring non-thermal plasma technology for microalgae removal

The global population and economic development surge has substantially increased water demand, resulting in heightened sewage and pollutant generation, posing environmental hazards. Addressing this challenge necessitates the implementation of efficient and cost-effective water reclamation methods. Non-thermal plasma technology (NTP) has emerged as a promising solution, garnering attention for its superior efficiency compared to alternatives. While existing studies have predominantly focused on energy efficiency and pollutant removal, limited research has delved into the biological removal aspect, particularly concerning algae. This study utilized a dielectric barrier plasma diffuser to eliminate Spirulina microalgae (Spirulina platensis) from wastewater solutions, demonstrating higher algae removal and superior mass transfer compared to alternative plasma methods. The effect of sample volume, input voltage and power, flow rate, and initial solution concentration on the algae removal was investigated. Investigation of operational parameters revealed the best condition resulting in a 98 % removal rate and 20 g/kWh energy efficiency. The best conditions for the removal of Spirulina microalgae were considered in a sample volume of 50 mL, a voltage of 7.6 kV, a flow rate of 700 mL/min, and an initial solution concentration of 1280 mg/liter. Scanning Electron Microscope (SEM) images illustrated the impact of active species on cell structure, leading to the destruction of spiral form and loss of reproductive ability. The study underscores the potential of NTP for efficient algae removal and identifies key active species involved in the process. The removal of Spirulina microalgae was attributed to a combination of singlet oxygen (1O2), hydroxyl radicals, and ozone.

Geometry of Modern Presbyopia-Correcting Intraocular Lenses and Changes in Haptic-Capsular Bag Behavior According to Compression and Different Well Diameters: A Bench Study Using Computed Tomography.

The geometry of an intraocular lens(IOL), the design of the haptics, and the optic-haptic junction play a role in initial and long-term visual outcome after cataract surgery. Knowledge of the behavior of an implant in the capsular bag and under compression is of major importance. Our laboratory experiment analyzed in-depth the geometry of acrylic, single-piece, premium, presbyopia-correcting intraocular lenses and changes in haptic-capsular bag relationships according to capsular bag size using a range of compression well diameters. One hydrophilic (RayOne Trifocal® RAO603F) and three hydrophobic intraocular lenses (AcrySof IQ PanOptix®, Synergy DFR00V™, and AT ELANA 841P®) were scanned with computed tomography (CT) in a dry, uncompressed state for quantitative analyses of haptic and optic-haptic junction (OHJ) dimensions and qualitative assessment of geometry. In the second part of the experiment, CT was performed after sample placement into a series of compression wells (9.0, 10.0, 11.0, 11.5mm) for analyses of length of contact (LoC) between the haptics and the wells. Axial alignment and haptic-capsular bag relationships were assessed. The qualitative and quantitative evaluations highlighted differences in haptic and OHJ geometry and dimensions across the samples. The mid-zone of the optic was thickest in the hydrophilic sample (RayOne Trifocal® RAO603F) with a maximum of 0.880mm compared to the thinnest hydrophobic sample (AT ELANA 841P®) with 0.564mm. The AT ELANA 841P® showed the largest OHJ surface area (3.86mm2) and OHJ volume (0.60mm3) of the hydrophobic samples. The TECNIS Synergy DFR00V™ showed the thickest OHJ (0.51mm), while the AcrySof IQ PanOptix® showed the thinnest OHJ (0.21mm). The LoC values decreased with increasing well size for all tested samples. The AT ELANA 841P® showed the largest LoC and largest contact zones of hydrophobic samples in all wells. The laboratory experiments highlight differences in the haptics, the OHJ geometric characteristics, and behavior of samples in different well diameters. The results support the idea that specific IOL designs may have advantages or disadvantages depending on anatomical dimensions. We cannot make any classification or rating (good versus bad) for clinical practice on the basis these experimental results, as many other factors play a role. However, knowledge of IOL geometry seems important to select the best option in each individual case.