A guide to selecting high-performing antibodies for Synaptotagmin-1 (Uniprot ID P21579) for use in western blot, immunoprecipitation, immunofluorescence and flow cytometry.

Synaptotagmin-1 is a synaptic vesicle transmembrane protein that senses calcium influx via its tandem C2-domains, triggering synchronous neurotransmitter release. Disruption to SYT1 is associated with neurodevelopmental disorders, highlighting the importance of identifying high-quality research reagents to enhance understanding of Synaptotagmin-1 in health and disease. Here we have characterized thirteen Synaptotagmin-1 commercial antibodies for western blot, immunoprecipitation, immunofluorescence and flow cytometry using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. These studies are part of a larger, collaborative initiative seeking to address antibody reproducibility issues by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs.

Enhanced multiferroic properties of NZCFO-BNFSO composites: Synthesis, characterization, and performance analysis.

The structural and multiferroic properties of xNi0.24Zn0.58Cu0.18Fe2O4(NZCFO)-(1-x)Bi0.9Nd0.1Fe0.95 Sc0.05O3(BNFSO) are explored in this paper. Bi2O3 additives significantly lower the sintering temperature of the composites. The XRD analysis validates the coexistence of hexagonal perovskite BNFSO and spinel NZCFO phases. The FESEM images illustrate an almost homogeneous amalgamation of the BNFSO and NZCFO grains. The real part of initial permeability and the relative quality factor increases with NZCFO contents in the composites. The maximum permeability is observed for the composite with 80 % ferrite content. The ferroelectric BNFSO exhibits antiferromagnetic behavior and with the increase in NZCFO the saturation magnetization increases significantly. The dielectric constant confirms typical dielectric dispersion at low frequencies because of Maxwell-Wagner space charge polarization. The P-E hysteresis measurement reveals that the composite with 40 % ferrite content exhibits the highest loop area and hence a large energy storage capacity. Incorporating BNFSO and NZCFO into the composite boosts the multiferroic properties, which might be a suitable alternative to single-phase multiferroics.

The development of bend clamp of carbon-based fiber offshore gas lift pipelines: A primary experimental approach.

The work reports the proposed development of the laminated carbon fiber-reinforced polymer composites (CFRP), at a uniaxial angle as a primary bend clamp material assembly of the offshore gas lift pipelines. Generally, the clamps are utilized to repair the defect of the metallic pipelines due to their practical and mechanical strength. The existing bend clamp model poses challenges especially when applied to the J-tube bending side due to mechanical buckling and corrosion. This study evaluates the effect of laminated CFRP subjected to uniform uniaxial and in-plane compression on API 5 L X52. Several tests, including tensile, bend, and impact tests, are chosen to assess the effect of multiple and thick carbon layers to improve the clamp's mechanical properties. Scanning electronic microscope (SEM) intends to reveal the surface characterization of carbon fiber clamp. The result shows that laminated carbon fiber increases the durability and mechanical properties of the bend clamp for two weeks. The tensile test result shows that the uniaxial orientation has the highest tensile strength of about 2000N and is affected by the strong interfacial bond between the fiber and metal layers. The high impact energy of 2242 ± 0.001 J of the same orientation shows the stability of the clamp under unprecedented dynamic load and enhances the safety. The SEM result at uniaxial shows the resistance towards fracture has increased and was initiated from the air pocket in the composite.

High hydrostatic pressure reduces inflammation induced by litchi thaumatin-like protein via altering active domain.

Thaumatin-like proteins (TLP), existing in various fruits, have allergenic and pro-inflammatory activities. The current research attempts to reduce the pro-inflammatory activity of litchi TLP (LcTLP) through high hydrostatic pressure (HHP). This study demonstrated that HHP (250-500 MPa, 5-10 min) was a potential technique to reduce the pro-inflammatory activity of LcTLP, which was attributed to the irreversible destruction of the active domain, ie., V-cleft. SDS-PAGE showed no change in the protein profile. Continuous HHP treatment promoted LcTLP unfolding and then reassembling (400 MPa was the transition pressure), and the content of ß-sheets decreased from 35.4% to 31.1%. HHP treatment could mitigate inflammatory responses of LcTLP, as confirmed by ELISA and western blot. Molecular dynamics simulations showed significant changes in the residue network under HHP, thereby affecting the V-cleft. These findings provide a theoretical explanation and structural insights into the HHP-induced reduction of pro-inflammatory activity of LcTLP.

Adaptively multi-scale microstructure characterization of cancellous bone via Photoacoustic signal decomposition.

Osteoporosis is a systemic disease with a high incidence in the elderly and seriously affects the quality of life of patients. Photoacoustic (PA) technology, which combines the advantages of light and ultrasound, can provide information about the physiological structure and chemical information of biological tissues in a non-invasive and non-radiative way. Due to the complex structural characteristics of bone tissue, PA signals generated by bone tissue are non-stationary and nonlinear. However, conventional PA signal processing methods are not effective for non-stationary signal processing. In this study, an empirical mode decomposition (EMD)-based Hilbert-Huang transform (HHT) PA signal analysis method, called HHT PA signal analysis (HPSA), was developed to assess the microstructure information of bone tissue, which is closely related to bone health. The feasibility of the HPSA method in bone health assessment was proven by numerical simulation and experimental studies on animal samples with different bone volume/total volume (BV/TV) and bone mineral densities. First, based on adaptive EMD, the different modes correlated with multi-scale information were mined from the PA signal, the correlations between different intrinsic mode function (IMF) modes and BV/TVs were analyzed, and the optimal mode for more efficient PA time-frequency analysis was selected. Second, multi-wavelength HPSA was used to assess the changes in the chemical components of the bone tissue. The results demonstrate that the HPSA method can distinguish bones with different BV/TVs and microstructure conditions adaptively with high efficiency. They further emphasize the potential of PA techniques in characterizing biological tissues in bones for early and rapid detection of bone diseases.

Structural characterization and immunomodulating assessment of ultra-purified water extracted fucoidans from Saccharina latissima, Alaria esculenta and Laminaria hyperborea.

Fucoidans, a group of high molecular weight polysaccharides derived mainly from brown algae, are characterized by their high fucose content, degree of sulfation (DS), and intra- and interspecific structural variation. Fucoidans are increasingly recognized due to various reported bioactivities, potentially beneficial for human health. To unlock their potential use within biomedical applications, a better understanding of their structure-functional relationship is needed. To achieve this, systematic bioactivity studies based on well-defined, pure fucoidans, and the establishment of standardized, satisfactory purification protocols are required. We performed a comprehensive compositional and structural characterization of crude and ultra-purified fucoidans from three kelps: Saccharina latissima (SL), Alaria esculenta (AE) and Laminaria hyperborea (LH). Further, the complement-inhibiting activity of the purified fucoidans was assessed in a human whole blood model. The purification process led to fucoidans with higher DS and fucose and lower concentrations of other monosaccharides. Fucoidans from SL and LH resembles homofucans, while AE is a heterofucan rich in galactose with comparably lower DS. Fucoidans from SL and LH showed complement-inhibiting activity in blood and blood plasma, while no inhibition was observed for AE under the same conditions. The results emphasize the importance of high DS and possibly fucose content for fucoidans' bioactive properties.

Investigation of thermal properties and structural characterization of novel boron-containing Schiff base polymers.

Three novel Schiff base polymers were synthesized by using the Schiff base monomer obtained using aldehyde containing boric acid and amines containing hydroxyl groups as the starting materials. The polymers were synthesized at the same temperature and using the same amount of oxidizing agent by the oxidative polycondensation method. The characterization of all polymers and monomers was achieved by using Fourier transform infrared spectroscopy (FT-IR), ultraviolet-visible spectroscopy (UV-vis), nuclear magnetic resonance spectroscopy (1H-NMR), liquid chromatography-mass spectrometry (LC-MS), gel permeation chromatography (GPC) and scanning electron microscopy (SEM). The thermal stability of these compounds was studied by employing thermogravimetric analysis (TGA), and thermodynamics parameters such as activation energy (Ea), enthalpy (∆H), entropy (∆S), and Gibbs free energy(∆G) for the decomposition process were calculated using the Flynn-Wall-Ozawa method.

Antibody characterization is critical to enhance reproducibility in biomedical research.

Antibodies are used in many areas of biomedical and clinical research, but many of these antibodies have not been adequately characterized, which casts doubt on the results reported in many scientific papers. This problem is compounded by a lack of suitable control experiments in many studies. In this article we review the history of the 'antibody characterization crisis', and we document efforts and initiatives to address the problem, notably for antibodies that target human proteins. We also present recommendations for a range of stakeholders - researchers, universities, journals, antibody vendors and repositories, scientific societies and funders - to increase the reproducibility of studies that rely on antibodies.

Development of a biomarker-based platform for comprehensive skin characterization using minimally invasive skin sampling and quantitative real-time PCR.

BACKGROUND: Classifying diverse skin types is crucial for promoting skin health. However, efficiently identifying and analyzing relevant biomarkers from a vast array of available genetic data is challenging. Therefore, this study aimed to develop a precise and efficient platform for analyzing specific skin biomarkers using quantitative real-time PCR (qRT-PCR) with the minimal invasive skin sampling method (MISSM). MATERIALS AND METHODS: MISSM was used for RNA extraction from skin samples, followed by qRT-PCR analysis to quantify the expression of 20 biomarkers associated with skin characteristics (four biomarkers each for five skin characteristics). Noninvasive measurements from 299 Korean participants were utilized to correlate biomarker expression with skin parameters. Statistical analyses were conducted between biomarker expression levels and noninvasive skin measurements to select the relatively best-performing biomarker for each skin characteristic. RESULTS: Collagen type 1 alpha 1 (COL1A1) and moesin (MSN) were identified as skin aging biomarkers. Krüppel-like factor 4 (KLF4) and serine peptidase inhibitor Kazal type 5 (SPINK5) were identified as skin dryness biomarkers, whereas melan-A (MLANA) was selected as a biomarker for understanding pigmentation dynamics. Myelin protein zero like 3 (MPZL3) and high mobility group box 2 (HMGB2) were identified as markers of oily skin and skin sensitivity, respectively. Statistically significant correlations were found between the biomarker expression levels and noninvasive skin characteristic measurements. CONCLUSION: This study successfully developed a platform for the precise evaluation of individual skin characteristics using MISSM and qRT-PCR biomarker analysis. By selecting biomarkers that correlate with noninvasive measurements of skin characteristics, we demonstrated the platform's efficacy in assessing diverse skin conditions.

Development of antimicrobial starch-based composite films reinforced with soybean expeller for sustainable active packaging applications.

In this study, the influence of glycerol and sonicated soybean expeller (SSE) on composite edible films supporting natamycin and nisin was investigated using Response Surface Methodology. Assessments were conducted on mechanical properties, moisture content, water solubility (SW), and color. Optimal results were achieved with 0.46% SSE and 1.4% glycerol, yielding a maximum tensile strength (TS) of 1.0 ± 0.1 MPa and a minimum SW of 19.0 ± 0.3%. SSE had no impact on Tg values (82-89 °C), while antimicrobials reduced Tg (70-73 °C) due to increased water retention. Water vapor permeability was (2.5 ± 0.2) × 10-9  -1 s-1 Pa-1. FTIR analysis revealed strong component interactions. The composite films demonstrated biodegradability in compost after seven days and effective action against Listeria innocua and Saccharomyces cerevisiae. These findings suggest that these materials hold promise as active films for food preservation. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01516-6.

Polycyclic aromatic hydrocarbons in fish from four lakes in central and eastern China: Bioaccumulation, pollution characteristics, sources, and health risk assessment.

Polycyclic aromatic hydrocarbons (PAHs) are highly concerning environmental pollutants due to their toxicity, persistence, and bioaccumulation. In this paper, concentrations and compositions of 16 United States Environmental Protection Agency (USEPA) priority control PAHs in the fish collected from four lakes in central and eastern China were analyzed. 18 species of fish were collected from four typical lakes, namely Taihu Lake, Danjiangkou Reservoir, Yuncheng Salt Lake, and Nansi Lake. Quantitative analysis of PAHs were carried out with gas chromatograph/mass spectrometer, and 13 out of 16 PAHs were identified, with the main components being pyrene, chrysene, naphthalene, and benzo(b)fluoranthene. The accumulation of PAHs in fish from Taihu Lake, Danjiangkou Reservoir, Yuncheng Salt Lake, and Nansi Lake was 28.75-47.27, 26.60-31.93, 33.56-39.30, and 27.22-43.01 ng·g-1, respectively. The toxic equivalents of high-cyclic PAHs in fish of the four lakes were significantly higher than those of low-cyclic and middle-cyclic PAHs (P < 0.05). In Taihu Lake, Danjiangkou Reservoir, and Nansi Lake, the toxicity equivalents were predominantly contributed by benzo[a] pyrene (BaP), while in Yuncheng Salt Lake, dibenzo(a,h) anthracene (DahA) was the main contributor. The residents in central and eastern China exposed to PAHs had a negligible non-cancer risk (non-carcinogenic risk values <1) and a potential low cancer risk. It was noteworthy that the Pleuronichthys cornutus and Lateolabrax japonicus from Yuncheng Salt Lake could pose carcinogenic risks (>10-4) to humans, with benzo[b]fluoranthene (BbF) having the highest risk contribution rate. Source analysis indicated that the main source of PAHs in fish was combustion sources. BaP, DahA, and BbF could become potential pollutants of concern in the field of ecotoxicology. The results of this study on PAHs bioaccumulation, pollution characteristics, sources and health risks in fish from four lakes would provide a scientific basis for local governments to formulate targeted environmental management policies, pollution control measures, and public health strategies.

Zn2+ chelating peptide GFLGSP: Characterization of structure/Zn2+ chelating mode and the potential mechanisms for promoting Zn2+ transport in Caco-2 cells.

This study focused on exploring the Zn2+ chelating peptide GFLGSP: the characterization of structure/Zn2+ chelating mode and the potential mechanisms for promoting Zn2+ transport in Caco-2 cells. The findings revealed the bidentate chelating between Zn2+ and carboxyl oxygen atom in Pro6 residue. Thereafter, the secondary structure of GFLGSP remained unchanged, but there was an increase in zeta potential and particle size. Notably, the GFLGSP-Zn2+ complex enhanced the Zn2+ transport rate and modulated ZIP4 and ZNT1 expression in a Caco-2 cells monolayer model. As revealed by molecular docking analysis, GFLGSP interacted with ZIP4 through intermolecular hydrogen bonds as well as Van der Waals forces. The Zn2+ transport mechanisms of the GFLGSP-Zn2+ complex encompassed ZIP4 (vital channel), endocytosis (primary pathway) and paracellular transport (supplementary pathway). Based on these results, the tilapia skin collagen-derived GFLGSP hold promise as the potential dietary Zn2+ supplement.

Onopordum platylepis (Murb.) Murb. as a novel source of thistle rennet: First application to the manufacture of traditional Italian raw ewe's milk cheese.

In this study, for the very first time, aqueous extracts obtained from flowers of spontaneously grown or cultivated Onopordum platylepis (Murb.) Murb. thistles were used as coagulating agents for the pilot-scale manufacture of Caciofiore, a traditional Italian raw ewe's milk cheese. Cheese prototypes were compared to control cheeses curdled with a commercial thistle rennet obtained from flowers of Cynara cardunculus L. After 45 days of ripening under controlled conditions, both the experimental and control cheese prototypes were analyzed for: cheese yield, physico-chemical (pH, titratable acidity, aw, proximate composition), morpho-textural (color and texture), and microbiological parameters (viable cell counts and species composition assessed by Illumina sequencing), as well as volatile profile by SPME-GC-MS. Slight variations in titratable acidity, color, and texture were observed among samples. Based on the results overall collected, neither the yield nor the proximate composition was apparently affected by the type of thistle coagulant. However, the experimental cheese prototypes curdled with extracts from flowers of both spontaneous or cultivated thistles showed 10 % higher values of water-soluble nitrogen compared to the control prototypes. On the other hand, these latter showed slightly higher loads of presumptive lactococci, thermophilic cocci, coliforms, and eumycetes, but lower counts of Escherichia coli. No statistically significant differences were revealed by the metataxonomic analysis of the bacterial and fungal biota. Though most volatile organic compounds (VOCs) were consistent among the prototypes, significant variability was observed in the abundance of some key aroma compounds, such as butanoic, hexanoic, and octanoic acids, ethanol, propan-2-ol, isobutyl acetate, 2-methyl butanoic acid, and 3-methyl butanal. However, further investigations are required to attribute these differences to either the type of coagulant or the metabolic activity of the microorganisms occurring in the analyzed cheese samples. The results overall collected support the potential exploitation of O. platylepis as a novel source of thistle coagulant to produce ewe's milk cheeses.

Seven sour substances enhancing characteristics and stability of whey protein isolate emulsion and its heat-induced emulsion gel under the non-acid condition.

Protein emulsion gels, as potential novel application ingredients in the food industry, are very unstable in their formation. However, the incorporation of sour substances (phosphoric acid, lactic acid, acetic acid, malic acid, glutamic acid, tartaric acid and citric acid) would potentially contribute to the stable formation of whey protein isolate (WPI) emulsion as well as its gel. Thus, in this work, physical stability of seven acid-treated WPI emulsions, and microstructures, rheological properties, water distribution of its emulsion gels were characterized and compared. Initially, the absolute zeta-potential, interfacial protein adsorption, and emulsifying characteristics of acid-induced WPI emulsions were higher in contrast to acid-untreated WPI emulsions. Moreover, acid-induced WPI emulsions were thermally induced (95 ℃, 30 min) to form its emulsion gel networks via disulfide bonds as the main force (acid-untreated WPI emulsions were unable to form gels). High-resolution microscopic observation revealed that acid-induced WPI in emulsion gel network showed the morphology of aggregates. Dynamic oscillatory rheology results indicated that acid-induced emulsion gel exhibited highly elastic behavior and its viscoelasticity was associated with the generation of protein gel networks and aggregates. In addition, PCA and heatmap results further illustrated that malic acid-induced WPI emulsion gels had the best water holding capacity and gel characteristics. Therefore, this study could provide an effective way for the foodstuffs industry to open up new texture and healthy emulsion gels as fat replaces and loading systems of bioactive substances.

Techniques for characterizing biofunctionalized surfaces for bioanalysis purposes.

Surface biofunctionalization is an essential stage in the preparation of any bioassay affecting its analytical performance. However, a complete characterization of the biofunctionalized surface, considering studies of coverage density, distribution and orientation of biomolecules, layer thickness, and target biorecognition efficiency, is not met most of the time. This review is a critical overview of the main techniques and strategies used for characterizing biofunctionalized surfaces and the process in between. Emphasis is given to scanning force microscopies as the most versatile and suitable tools to evaluate the quality of the biofunctionalized surfaces in real-time dynamic experiments, highlighting the helpful of atomic force microscopy, Kelvin probe force microscopy, electrochemical atomic force microscopy and photo-induced force microscopy. Other techniques such as optical and electronic microscopies, quartz crystal microbalance, X-ray photoelectron spectroscopy, contact angle, and electrochemical techniques, are also discussed regarding their advantages and disadvantages in addressing the whole characterization of the biomodified surface. Scarce reviews point out the importance of practicing an entire characterization of the biofunctionalized surfaces. This is the first review that embraces this topic discussing a wide variety of characterization tools applied in any bioanalysis platform developed to detect both clinical and environmental analytes. This survey provides information to the analysts on the applications, strengths, and weaknesses of the techniques discussed here to extract fruitful insights from them. The aim is to prompt and help the analysts to accomplish an entire assessment of the biofunctionalized surface, and profit from the information obtained to enhance the bioanalysis output.

Equivalent Electrical Circuit Approach to Enhance a Transducer for Insulin Bioavailability Assessment.

The equivalent electrical circuit approach is explored to improve a bioimpedance-based transducer for measuring the bioavailability of synthetic insulin already presented in previous studies. In particular, the electrical parameter most sensitive to the variation of insulin amount injected was identified. Eggplants were used to emulate human electrical behavior under a quasi-static assumption guaranteed by a very low measurement time compared to the estimated insulin absorption time. Measurements were conducted with the EVAL-AD5940BIOZ by applying a sinusoidal voltage signal with an amplitude of 100 mV and acquiring impedance spectra in the range [1-100] kHz. 14 units of insulin were gradually administered using a Lilly's Insulin Pen having a 0.4 cm long needle. Modified Hayden's model was adopted as a reference circuit and the electrical component modeling the extracellular fluids was found to be the most insulin-sensitive parameter. The trnasducer achieves a state-of-the-art sensitivity of 225.90 ml1. An improvement of 223 % in sensitivity, 44 % in deterministic error, 7 % in nonlinearity, and 42 % in reproducibility was achieved compared to previous experimental studies. The clinical impact of the transducer was evaluated by projecting its impact on a Smart Insulin Pen for real-time measurement of insulin bioavailability. The wide gain in sensitivity of the bioimpedance-based transducer results in a significant reduction of the uncertainty of the Smart Insulin Pen. Considering the same improvement in in-vivo applications, the uncertainty of the Smart Insulin Pen is decreased from [Formula: see text]l to [Formula: see text]l.Clinical and Translational Impact Statement: A Smart Insulin Pen based on impedance spectroscopy and equivalent electrical circuit approach could be an effective solution for the non-invasive and real-time measurement of synthetic insulin uptake after subcutaneous administration.

Plant-Based Synthesis, characterization Approaches, Applications and Toxicity of Silver Nanoparticles: A Comprehensive Review.

The development of an environmentally benign method for the synthesis of nanoparticles has been facilitated by green chemistry. "Green synthesis" uses a range of biological elements like microbes, plants, and other biodegradable materials to produce NPs. Active biomolecules that are secreted by natural strains and present in the plant extracts serve as both reducing and capping/stabilizing agents. Microorganisms' intracellular enzymes can reduce metal ions, which explains how NPs might potentially nucleate. Plant-based synthesis of nanomaterials is particularly promising owing to abundant resources, simplicity of synthesis, and low cost. Silver nanoparticles (AgNPs) are attracting great attention in the research community due to their wide variety of applications in chemistry, food technology, microbiology, and biomedicine. Recent years have seen a large amount of research on the bio-genic synthesis of AgNPs employing biomaterials like plant extract and bacteria as reducing agents. Herein we discuss a thorough overview of the plant-based synthesis of silver nanoparticles (AgNPs), characterization approaches, applications, and toxicity. The review covers the green chemistry and nanotechnology elements of producing AgNPs, including a thorough discussion of the plant extract mediated synthesis, detailed formation mechanism, and a well-balanced emphasis on hazards and advantages. Based on current developments, the optimisation strategies, applications, and interdisciplinary characteristics are also covered in detail.

Risky lane-changing behavior recognition based on stacking ensemble learning on snowy and icy surfaces.

Risky lane-changing (LC) behavior adversely affects traffic safety, especially on snowy and icy surfaces. However, due to the particularity of the snowy and icy surfaces and the scarcity of data, research on risky lane-changing behavior (RLCB) under extreme scenarios is insufficient. Therefore, this study presents a novel research framework aimed at selecting key risk characterization indicators (RCIs) and identifying RLCB on highways using driving simulation data on snowy and icy surfaces. A highway LC scenario was established on snowy and icy surfaces using a driving simulator, and 1200 sets of LC sample data were extracted. From the perspectives of parameter importance and correlation, 12 key RCIs with high importance and low inter-correlation were selected using the C4.5 decision tree algorithm and Pearson correlation analysis method. The RLCB recognition model was developed using the Stacking ensemble learning method and then compared with traditional recognition algorithms. The results show that the accuracy of the recognition model based on the Stacking ensemble learning model is significantly better than that of traditional algorithms, with a recognition accuracy of 98.33%. This finding can provide the basis for developing LC warning systems for intelligent connected vehicles on snowy and icy surfaces.

Comparison of the neurotoxic potency of different ultrafine particle fractions from diesel engine exhaust following direct and simulated inhalation exposure.

Exposure to traffic-related air pollution and ultrafine particles (<100 µm; UFP) is linked with neurodegeneration. However, the impact of the aromatic content in fuels and the contribution of different fractions of UFP, i.e., solid UFP vs SVOC UFP, on neuronal function is unknown. We therefore studied effects on neuronal activity and viability in rat primary cortical cells exposed for up to 120 h to copper oxide particles (CuO) or UFP (solid and SVOC) emitted from a heavy-duty diesel engine fueled with petroleum diesel (A20; 20 % aromatics) or Hydrotreated Vegetable Oil-type fuel (A0; 0.1 % aromatics), or solid UFP emitted from a non-road Kubota engine fueled with A20. Moreover, effects of UFP and CuO upon simulated inhalation exposure were studied by exposing an lung model (Calu-3 and THP-1 cells) for 48 h and subsequently exposing the cortical cells to the medium collected from the basal compartment of the lung model. Additionally, cell viability, cytotoxicity, barrier function, inflammation, and oxidative and cell stress were studied in the lung model after 48 h exposure to UFP and CuO. Compared to control, direct exposure to CuO and SVOC UFP decreased neuronal activity, which was partly associated with cytotoxicity. Effects on neuronal activity upon direct exposure to solid UFP were limited. A20-derived UFP (solid and SVOC) were more potent in altering neuronal function and viability than A0 counterparts. Effects on neuronal activity from simulated inhalation exposure were minor compared to direct exposures. In the lung model, CuO and A20-derived UFP increased cytokine release compared to control, whereas CuO and SVOC A20 altered gene expression indicative for oxidative stress. Our data indicate that SVOC UFP exhibit higher (neuro)toxic potency for altering neuronal activity in rat primary cortical cells than the solid fraction. Moreover, our data suggest that reducing the aromatic content in fuel decreases the (neuro)toxic potency of emitted UFP.

Immune profiling and functional analysis of NK and T cells in ataxia telangiectasia.

Ataxia telangiectasia (AT) is a rare autosomal-recessive disorder characterized by profound neurodegeneration, combined immunodeficiency, and an increased risk for malignant diseases. Treatment options for AT are limited, and the long-term survival prognosis for patients remains grim, primarily due to the emergence of chronic respiratory pathologies, malignancies, and neurological complications. Understanding the dysregulation of the immune system in AT is fundamental for the development of novel treatment strategies. In this context, we performed a retrospective longitudinal immunemonitoring of lymphocyte subset distribution in a cohort of AT patients (n = 65). Furthermore, we performed FACS analyses of peripheral blood mononuclear cells from a subgroup of 12 AT patients to examine NK and T cells for the expression of activating and functional markers. We observed reduced levels of peripheral blood CD3+CD8+ cytotoxic T cells, CD3+CD4+ T helper cells, and CD19+ B cells, whereas the amount of CD3--CD56+ NK cells and CD3+CD56+ NKT-like cells was similar compared with age-matched controls. Notably, there was no association between the age-dependent kinetic of T-, B-, or NK-cell counts and the occurrence of malignancy in AT patients. Additionally, our results indicate an altered NK- and T-cell response to cytokine stimulation in AT with increased levels of TRAIL, FasL, and CD16 expression in NK cells, as well as an elevated activation level of T cells in AT with notably higher expression levels of IFN-γ, CD107a, TRAIL, and FasL. Together, these findings imply function alterations in AT lymphocytes, specifically in T and NK cells, shedding light on potential pathways for innovative therapies.