Absolute Amount Research Articles

Herbivore grazing enhances macroalgal organic carbon release and alters their carbon sequestration fate in the ocean

Herbivore grazing on macroalgae promotes the release of macroalgal organic carbons into seawater and potentially impacts their bioavailability. However, the influence of herbivores on the fate of macroalgal organic carbon remains unclear, hindering a comprehensive and in-depth understanding of the role of macroalgae in ocean carbon cycle. Here, we cocultured suspended herbivore (Apohyale sp.) and benthic herbivore (Nereis diversicolor) with macroalgae (Ulva prolifera) in the laboratory, and found that the two grazers promote the release of macroalgal organic carbon through different pathways. Apohyale sp. can simultaneously increase the release of different forms of organic carbon by feeding on U. prolifera thalli, including dissolved organic carbon (DOC), particluate organic carbon (POC), and algal organic detritus; while N. diversicolor demonstrated a preference for ingesting algal detritus and POC, thereby reducing the detrital carbon but greatly promoting their conversion to DOC. The amount of organic carbon released per day after predation by Apohyale sp. is much higher (7.2 vs 0.5 mg C d-1) than by N. diversicolor. Meanwhile, through long-term microbial degradation experiments, we found that herbivores significantly alter the fate of macroalgae organic carbon. Although the proportions of stable carbon (recalcitrant DOC and recalcitrant POC) in different forms of macroalgal organic carbon varied after predation, the absolute amount of their residuals in seawater were 2-3 times higher than those not ingested by herbivores. Our results highlight that herbivores play a pivotal role in promoting carbon flow in marine food webs and have a significant impact on macroalgal carbon sequestration.

Counteracting the Loss of Release for Indomethacin-Copovidone ASDs

This work revisits the changing release behavior of indomethacin(IND)-copovidone amorphous solid dispersions (ASDs) when increasing their drug load (DL). While showing congruent release behavior at DL 0.1, ASDs with DLs of 0.3 and higher show incongruent release finally resulting in a complete loss of release. To study and explain this phenomenon, we modeled the release kinetics of these ASDs and looked into their phase behavior both experimentally and theoretically. We applied a diffusion model to accurately describe experimental release profiles for congruent release, incongruent release as well as for loss of release. Predicted concentration profiles for IND, copovidone, and water within the ASD revealed the formation of an ASD layer that almost exclusively contains amorphous IND. Our phase-diagram predictions and experimental data explain this phenomenon by water-induced phase separation in those parts of the ASD which did absorb water from the dissolution medium. Whereas the evolving copovidone-rich phase dissolved, the IND-rich phase remained undissolved and formed a super-hydrophobic cover of the remaining inner core of the ASD, thus finally completely preventing its dissolution. Higher DLs promote phase separation. This leads to the counterintuitive effect that the higher the DL, the lower the absolute amount of IND released. While the ASD containing 6 mg IND (DL 0.1) released 6 mg IND, the one containing 42 mg IND (DL 0.7) released only 1 mg IND. The theoretical approach applied in this work is for the first time able to quantitatively predict that reducing DL or tablet size could be used to overcome this problem.

Populating HIA of tobacco policies with empirical evidence on behavioral response to price increases

Abstract Performing Health Impact Assessment of tobacco policies requires to have insights into how smokers will react to policy measures such as price increases. In the Netherlands, a series of 3 substantial price increases has been implemented in 2020 (10% increase), 2023 (10% increase) and 2024 (24% increase). We performed three consecutive studies comparing the ex-ante stated preferences about behavioural response to actual and hypothetical future price increases, and compared these stated preferences with actual behaviour changes 6 months after the increase in excise tax. Two of these possible behavioural reactions actually will impact smoking prevalence or smoking intensity, e.g. stopping altogether and decreasing the number of cigarettes smoked. Other behavioural responses are less favourable and will not impact health, such as buying more and cheaper tobacco abroad and smoking less expensive brands or transferring from cigarettes to roll-your-own or e-cigarettes. For the price increases of 2020 and 2023, we have observed a substantial number of about 10% of smokers quitting successfully and about one third attempting to quit smoking. (Attempting to) quit smoking was much more prevalent among those with a more pronounced intention to stop smoking at the ex-ante measurements. At the same time, cross-border shopping increased significantly as well, with a 10 percent-point growth in the absolute amount of all tobacco consumed originating from abroad following the 10 percent price increase of April 2023. This big increase in cross-border shopping appeared to be independent of distance to the border. We conclude that every significant price increase stimulates smokers to consider to stop smoking. However, unilateral price increases in one country will eventually also stimulate smokers to buy cross-border. Behavioural data as presented should inform HIA to make them more realistic with regard to actual behaviour changes and health impacts of smoking policies.

Impact of waist circumference and abdominal fat distribution on coronary artery disease severity stratified by computed tomography-derived SYNTAX score

Abstract Background Waist circumference (WC) is the most convenient anthropometric measure of obesity. Recently, abdominal fat distribution (AFD) has been identified as a more important risk factor for coronary artery disease (CAD) than the absolute amount of adipose tissue. However, studies examining the impact of body composition indicators (BCIs) on CAD severity are lacking. The computed tomography (CT)-derived SYNTAX score (CT-SXscore) is a feasible method for grading CAD severity based on coronary CT angiography (CTA) findings. Purpose This study was aimed to evaluate the association between BCI and CAD severity using the CT-SXscore. Methods We enrolled consecutive patients with suspected CAD who underwent CTA using a 320-row multidetector CT scanner between October 2014 and March 2020. Plain abdominal CT was also performed at the umbilical level to measure the visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) areas. The total adipose tissue (TAT) area was calculated as the sum of VAT and SAT. To assess the AFD, the VAT/SAT (V/S) ratios were also calculated as the VAT area divided by the SAT area in each case. WC was measured by tracing the body contour at the umbilical level in CT images. The severity of coronary artery stenosis was assessed using CTA. Significant stenosis was defined as a stenosis diameter of ≥50%. The CT-SXscore was calculated in patients with more than one significant stenoses. Each stenotic lesion was evaluated to calculate the CT-SXscore in the same manner as the invasive coronary angiography assessment. We assessed the effects of BCI [i.e., WC, body mass index (BMI), each adipose tissue area, and V/S ratios] on the CT-SXscores. Results 308 eligible patients (mean age, 70.9 ± 10.6 years; 69.5% males) were evaluated. The median values of WC, BMI, TAT, SAT, VAT, and V/S ratio were 84.8 cm, 23.6 kg/m2, 198.2 cm2, 111.8 cm2, 81.0 cm2, and 0.67, respectively. Although WC showed a significant correlation with BMI, TAT, SAT, and VAT area, no correlation was observed between WC and V/S ratio (Figure1). The CT-SXscore showed no association with WC, BMI, and TAT area; however, it exhibited a weak negative correlation with the SAT area and a weak positive correlation with the VAT area. Additionally, a strong correlation was observed between the CT-SXscore and V/S ratio. In the multivariate regression analysis, WC, BMI, TAT, SAT, and VAT were no longer associated with the CT-SXscore after adjusting for traditional coronary risk factors, whereas the V/S ratio remained as the only independent predictor of CAD severity based on the CT-SXscore (Figure2). Conclusions WC was strongly correlated with other BCIs, except for the V/S ratio; however, it could not be identified as a predictive factor of CAD severity. Among the BCIs, only the V/S ratio was an independent predictor of CAD severity, indicating that the AFD revealed by the V/S ratio may be a more important risk factor for progression of CAD than other BCIs.Figure1Figure2

Study on the variability of oxygen adsorption behavior in coal gangue based on pore size structure

To investigate the adsorption and migration behavior of oxygen in coal gangue with different pore sizes, this study characterizes the multiscale morphology of coal gangue pores and fractures using SEM NMR, and low-temperature N2 (77 K) adsorption methods. As well, the study simulates the adsorption and diffusion behavior of O2 within the pore structures of coal gangue based on GCMC and MD methods. The study examines the oxygen adsorption characteristics across different pore sizes and analyzes the mass transfer mechanisms during oxygen diffusion. Results indicate that the pores in coal gangue are predominantly micropores within the 1–10 nm range. As pore size increases, the absolute adsorption amount of oxygen increases while the isosteric heat of adsorption decreases. The isosteric heat of adsorption in coal gangue models with different pore sizes is less than 42 kJ/mol, indicating the physical adsorption of oxygen within the pores. Oxygen shows adsorption layering perpendicular to the coal gangue surface, with adsorption density distribution differences decreasing and adsorption capacity weakening as pore size increases. The O2 adsorption isotherms in coal gangue follow Langmuir adsorption principles, with oxygen saturation adsorption decreasing as absolute adsorption increases. The confinement effect of coal gangue on oxygen weakens as pore size increases, and the impact of increasing pore size on oxygen diffusion is stronger than adsorption. These findings are significant for understanding the microscopic adsorption and diffusion behavior of O2 in coal gangue pores and fractures and are crucial for accurately preventing and managing spontaneous combustion in coal gangue piles.

Analysis of immunopathogenesis of psoriasis in the dynamics of treatment

Оbjective — to analyze the parameters of the immune system of the organism in patients with psoriasis before and after treatment. Materials and methods. The parameters of cellular immunity in an organism estimated after the change of relative and absolute amount of T- (CD3+) and B- (CD19+) of lymphocytes and subpopulations (CD4+, CD8+, CD16+, CD25+, CD71+, CD30+, CD95+). To identify of surface structures of lymphocytes the method of direct immune fluorescence was used where fluorescent label was attached to anti-CD of mononuclear antibodies of series Leu of the firm «Becton Dikinson» (USA). Calculations were conducted on the laser instantaneous cytofluorometer of the firm «Baston» (USA). Results and discussions. The most positive influence on the parameters of the cellular link of the immune system of the organism was found in patients with psoriasis who received immunobiological therapy with the medication etanercept in combination with narrowband (311 nm) phototherapy. After treatment in patients of this group, due to a significant decrease in the production of TNF and its serum concentration, the pro-inflammatory changes in the cellular link of the immune system decreased, the number of lymphocytes with the an early activation marker, which initiated a further cytokine cascade of development and prolongation of inflammation decreased significantly. In addition, the established reduction in the number of CD30+ lymphocytes is an indirect factor indicating the switching of the Tx2 response (autoimmune manifestations) to Tx1, and hence the achievement of the clinical and immunological remission of the psoriatic process. It is extremely important for the functioning of the immune system the decrease in the content of activated lymphocytes that express the Fas-receptor as due to the increased lymphocyte apoptosis, namely, T-cytotoxic lymphocytes, and autoimmune and proliferative changes took place inherent in the exacerbation of the psoriatic process. Conclusions. It was established that all our schemes of psoriasis treatment have immunorehabiting properties but their degree of severity was different. The most effective treatment scheme is the combined use of etanercept and UVB therapy, thus proliferative and inflammatory changes in the skin reduced, antigenic loading decreased, the level of CICs, autosensibilization and autoimmune disorders also decreased.

Whole-brain annotation and multi-connectome cell typing of Drosophila

The fruit fly Drosophila melanogaster has emerged as a key model organism in neuroscience, in large part due to the concentration of collaboratively generated molecular, genetic and digital resources available for it. Here we complement the approximately 140,000 neuron FlyWire whole-brain connectome1 with a systematic and hierarchical annotation of neuronal classes, cell types and developmental units (hemilineages). Of 8,453 annotated cell types, 3,643 were previously proposed in the partial hemibrain connectome2, and 4,581 are new types, mostly from brain regions outside the hemibrain subvolume. Although nearly all hemibrain neurons could be matched morphologically in FlyWire, about one-third of cell types proposed for the hemibrain could not be reliably reidentified. We therefore propose a new definition of cell type as groups of cells that are each quantitatively more similar to cells in a different brain than to any other cell in the same brain, and we validate this definition through joint analysis of FlyWire and hemibrain connectomes. Further analysis defined simple heuristics for the reliability of connections between brains, revealed broad stereotypy and occasional variability in neuron count and connectivity, and provided evidence for functional homeostasis in the mushroom body through adjustments of the absolute amount of excitatory input while maintaining the excitation/inhibition ratio. Our work defines a consensus cell type atlas for the fly brain and provides both an intellectual framework and open-source toolchain for brain-scale comparative connectomics.

Comparison of water quality, planktonic community, and volatile organic compounds in the seawater from five cage culture areas of large yellow croaker

The rapid development of marine fish cage farming has led to ecological damage, adversely affecting fish quality. Significant regional quality differences in large yellow croaker under similar farming conditions suggest that the water environment plays a crucial role in fish quality. Using full-length18s/16 s rRNA sequencing with bacterial absolute quantity and headspace solid-phase microextraction coupled with gas chromatography mass spectrometry (HS-SPME-GC–MS), we investigated the planktonic community and volatile organic compounds (VOCs) in seawater from cages housing large yellow croaker at five major mariculture sites in China, including ND (26.627°E, 119.638°N), NJ (27.479°E, 121.052°N), DT (27.981°E, 121.196°N), NC (28.470°E, 121.907°N), and DJ (30.198°E, 122.698°N). Notably, three dominant ciliates, Mesodinium rubrum, Strombidium sp., and Pseudotontonia simplicidens, were identified in NJ as bioindicators for water quality. Additionally, NJ cages had the highest bacterial biomass with 54 % Synechococcus CC9902, and the lowest abundance of bacteria with pathogenic activities. These may explain NJ's optimal water quality parameters and the highest eukaryotic biodiversity. In contrast, DT and DC contained only the core ciliate M. rubrum, with no dominant ciliate in ND. Styela gibbsii, a filter feeder, predominated in ND cages, adhering to the net. The occurrence of red tide-causing species and pathogenic bacteria or fungi was notable in DJ, DC, DT, and ND, including Akashiwo sanguinea, an unclassified Dinophyceae, Gyrodinium fusiforme, Pirum gemmate, Candida parapsilosis, Hortaea werneckii, and Pseudomonas juntendi. Bacterial functional analysis showed that ND cages harbored unique strains linked to ureolysis and various pathogenic activities. Co-association networks suggested that 2,4-Di-tert-butylphenol and pentadecane might play roles in communication within plankton communities. A rise in pleasant VOCs such as acetyl valeryl, pentadecane, butyl butyrate, tetradecane, and 5-butyldihydro-2(3H)-furanone was observed in DJ, DC, DT, and NJ, compared to ND. The results provide valuable insights for improving aquaculture practices and mitigating environmental impacts. The results provide valuable bioindicators for monitoring water quality using microbial communities and VOC profiles.

Be bold, start cold! cold formalin fixation of colorectal cancer specimens granted superior DNA and RNA quality for downstream molecular analysis

The use of cold formalin fixation (CFF; i.e., fixating tissue samples with 4 °C precooled formalin) recently attracted further attention owing to its putative improved ability to preserve nucleic acid compared with standard room temperature formalin (SFF). In this study, we aimed to assess the effect of four formalin-based fixation protocols (SFF, CFF, delayed formalin fixation-DFF, and cold formalin hyperfixation; CFH) on both DNA and RNA quality. We collected 97 colorectal cancer (CRC) and analyzed 23 metrics of nucleic acid quantity and quality yield using a multiplatform approach by combining spectrophotometric, fluorimetric, electrophoretic, and polymerase chain reaction (PCR) assays. Following confirmation of fixation-protocol-related different effects via clustering analysis, CFF presented best metrics compared with all protocols, specifically positive coefficients of DV1000-60000, DV2/DV1, DNA λ ratio 260/230, and ABL gene expression absolute copies, and negative coefficient of DV150-1000. The SFF subgroup presented a positive coefficient of DV150-1000 and negative coefficients for DV1000-60000, DV2/DV1, RNA λ ratio 260/230, RNA QuBit concentration, DV100/200, RNA electrophoresis concentration and absolute quantity, and ABL copies. Overall, we confirmed the superior yield performances of CFF preservation for both DNA and RNA compared with the other protocols in our series of CRC samples. Pending further validations and clarification of the specific mechanisms behind these findings, our study supports the implementation of CFF in the pathology unit routine specimen management for tumor tissue molecular profiling.

A gender and size specific evaluation of Grammont-type inlay versus lateralizing onlay stem designs in achieving lateralization and distalization in reverse shoulder arthroplasty

IntroductionIn reverse shoulder arthroplasty (RSA) new designs enable greater amounts of lateralization to prevent instability and scapular notching and increase range of motion, however, excessive lateralization leads to stress upon the acromion that can result in scapular spine fatigue fractures. Aim of this study was to gender- and size-specifically assess the influence of glenosphere size and different humeral designs on lateralization, distalization, and bony impingement-free range of motion (ROM) in patients undergoing RSA.MethodsComputed tomography scans from 30 osteoarthritic patients (f:15, m:15) and 20 cuff tear arthropathy patients (f:10, m:10) were used to virtually simulate RSA implantation. The efficacy of an inlay Grammont-type system vs. an onlay lateralizing system combined with different glenosphere sizes (36 mm vs. 42 mm) in achieving ROM, lateralization, and distalization was evaluated. Moreover, gender and patient’s constitution were correlated to humeral size by radiologically measuring the best-fit circle of the humeral head.ResultsA different amount of relative lateralization was achieved in both genders using large glenospheres and onlay designs. Latter yielded a higher ROM in all planes for men and women with a 42 mm glenosphere; with the 36 mm glenosphere, an increased ROM was observed only in men. The 155° inlay design led to joint medialization only in men, whereas all designs led to lateralization in women. When adjusting the absolute amount of lateralization to humerus’ size (or patient’s height), regardless of implant type, women received greater relative lateralization using 36 mm glenosphere (inlay: 1%; onlay 12%) than men with 42 mm glenosphere (inlay: -3%; onlay: 8%).ConclusionThe relative lateralization achieved using onlay design is much higher in women than men. Small glenospheres yield greater relative lateralization in women compared to large glenospheres in men. Humeral lateralization using onlay designs should be used cautiously in women, as they lead to great relative lateralization increasing stress onto the acromion.Level of evidenceBasic Science Study, Computer Modeling.

The compartment-specific manipulation of the NAD+/NADH ratio affects the metabolome and the function of glioblastoma

Glioblastoma multiforme (GBM) is the most aggressive type of cancer in the brain and has an inferior prognosis because of the lack of suitable medicine, largely due to its tremendous invasion. GBM has selfish metabolic pathways to promote migration, invasion, and proliferation compared to normal cells. Among various metabolic pathways, NAD (nicotinamide adenine dinucleotide) is essential in generating ATP and is used as a resource for cancer cells. LbNOX (Lactobacillus brevis NADH oxidase) is an enzyme that can directly manipulate the NAD+/NADH ratio. In this study, we found that an increased NAD+/NADH ratio by LbNOX or mitoLbNOX reduced intracellular glutamate and calcium responses and reduced invasion capacity in GBM. However, the invasion was not affected in GBM by rotenone, an ETC (Electron Transport Chain) complex I inhibitor, or nicotinamide riboside, a NAD+ precursor, suggesting that the crucial factor is the NAD+/NADH ratio rather than the absolute quantity of ATP or NAD+ for the invasion of GBM. To develop a more accurate and effective GBM treatment, our findings highlight the importance of developing a new medicine that targets the regulation of the NAD+/NADH ratio, given the current lack of effective treatment options for this brain cancer.

Benchtop IR Imaging of Live Cells: Monitoring the Total Mass of Biomolecules in Single Cells.

Absolute quantity imaging of biomolecules on a single cell level is critical for measurement assurance in biosciences and bioindustries. While infrared (IR) transmission microscopy is a powerful label-free imaging modality capable of chemical quantification, its applicability to hydrated biological samples remains challenging due to the strong IR absorption by water. Traditional IR imaging of hydrated cells relies on powerful light sources, such as synchrotrons, to mitigate the light absorption by water. However, we overcome this challenge by applying a solvent absorption compensation (SAC) technique to a home-built benchtop IR microscope based on an external-cavity quantum cascade laser. SAC-IR microscopy adjusts the incident light using a pair of polarizers to precompensate the IR absorption by water while retaining the full dynamic range. Integrating the IR absorbance over a cell yields the total mass of biomolecules per cell. We monitor the total mass of the biomolecules of live fibroblast cells over 12 h, demonstrating promise for advancing our understanding of the biomolecular processes occurring in live cells on the single-cell level.

57 Association of Absolute Amount of Pattern 4 Disease on Prostate Biopsy with Oncologic Outcomes in Intermediate-Risk Prostate Cancer: A Systematic Review

57 Association of Absolute Amount of Pattern 4 Disease on Prostate Biopsy with Oncologic Outcomes in Intermediate-Risk Prostate Cancer: A Systematic Review

Moderating Effects of Visual Order in Graphical Symbol Complexity: The Practical Implications for Design

In the field of visual graphic design, complexity plays a crucial role in visual information processing, and it is assumed to be an absolute quantity based on the number of the presenting features and components. However, it remains unclear whether the visual order of the constituent elements in graphical symbol complexity affects cognitive processing, especially memory processing. Our research innovatively generated four groups of novel, meaningless graphical symbols (complex and ordered, complex and disordered, simple and ordered, and simple and disordered) and experimentally manipulated the level of complexity and order in these stimuli. Before the formal experiment, a five-point scale was used to further rule out differences between objective and subjective definitions of these graphical symbols on ratings of complexity, order, concreteness, and familiarity. Then, we used a cue-recall task to compare subjects’ memory performance of those four graphical symbol groups. The analytical results showed a significant interaction between visual order and graphical symbol complexity, with the complexity effect appearing only when the stimuli were in disordered condition and disappearing once the stimuli were ordered. In addition, this study conducted a practical application validation to confirm that increasing the level of visual order is an effective way to improve user experience while maintaining the same level of complexity. The findings can serve as a reference for graphical symbol design, graphic design, and visual communication design.

Distribution Control and Environmental Fate of PFAS in the Offshore Region Adjacent to the Yangtze River Estuary─A Study Combining Multiple Phases Analysis.

The Yangtze River Estuary is the terminal sink of terrestrial per- and polyfluoroalkyl substances (PFAS) from the Yangtze River, while the environmental fate characteristics of legacy and emerging PFAS around this region have rarely been discussed. Here, 24 targeted PFAS in seawater, sediments, suspended particulate matter (SPM), and plankton in the offshore region adjacent to this estuary were investigated. The three dominant PFAS in all phases were perfluorooctanoic acid (PFOA, 23.8-61.9%), perfluorobutanoic acid (PFBA, 23.6-42.8%), and perfluoro(2-methyl-3-oxahexanoic) acid (HFPO-DA, 6.1-12.1%), and perfluoro-1-butane sulfonamide (FBSA, 0.1-7.3%) was first detected. The horizontal distributions of PFAS were dependent on salinity and disturbed by multiple water masses, while the vertical variations could be explained by their different partitioning characteristics in the water-SPM-sediment system (partition coefficients, Log Kd and Log Koc) and plankton (bioaccumulation factors, Log BAF). Although physical mixing was the major driver for PFAS settling (>83.7%), the absolute settling amount caused by the biological pump was still high (150.00-41994.65 ng m-2 day-1). More importantly, we found unexpected high Log Kd values of PFBA (2.24-4.55) and HFPO-DA (2.26-4.67), equal to PFOA (2.28-4.72), which brought concerns about their environmental persistence. Considering the increased detection of short-chain and emerging PFAS, more comprehensive environmental behaviors analysis is required urgently.

Возрастная динамика субпопуляций Т-лимфоцитов у пациентов с синдромом делеции 22q11.2 (синдромом Ди Джорджи)

Patients with the 22q11.2 deletion syndrome (DiGeorge syndrome, DDS) suffer from T-cell lymphopenia, changes in function and subpopulation of T-lymphocytes. The bibliographical data conflicts on the characteristics of T-lymphocytes in patients with DDS and their changes during physiological maturation. The purpose of this research was to follow-up patients with emphasis on the dynamics of changes in T-lymphocyte subpopulations with age. Methods used: 117 patients observed during 2013-2023 who were administered at the Allergology and Immunology Department with the G.N. Speransky City Children’s Hospital No. 9 (Moscow, Russia) with a diagnosis of DDS and who had undergone blood sampling to assess T-lymphocyte subpopulations using flow cytometry. All patients (0 to 18 y/o) were divided into 4 age groups: 0 to 2 y/o, 2 to 5 y/o, 5 to 10 y/o and 10 to 18 y/o. In parallel, the blood samples of 185 apparently healthy children of the corresponding age were examined. Results: a decrease in CD3 T-lymphocytes, CD4 T-helper cells, CD8 T-cytotoxic lymphocytes, CD4 early thymic emigrants and CD4 naive T-lymphocytes was found in all age groups (p<0.05). CD8+ naive T-lymphocytes and CD8+ early thymic emigrants were reduced in all age groups, but in the 2 to 5 y/o age group their values were close to normal (p=0.072 and p=0.220, respectively). CD4+ central memory cells were elevated in all age groups (p<0.001), while CD8+ central memory cells, CD4+ and CD8+ effector memory cells differed in that they had normal values in the 2 to 5 y/o age group (p=0.229, p=0.457 and p=0.140, respectively). CD4+ TEMRA were significantly increased in the 0 to 2 y/o age group (p=0.002), and in older age groups they tended to normal values. CD8+ TEMRA did not differ from the control group and did not have age-related dynamics. The T-helper subpopulations were characterized by a significant increase in the percentage of T-helper type 1, T-helper 17, T-helper 17.1 and a decrease in T-helper type 2 compared to the population of healthy blood sample donors (p=0.001), except for patients from the 2 to 5 y/o age group. T-regulatory cells in absolute amount were reduced in all age groups (p<0.001), while their relative number corresponded to the norm in the 0 to 2 y/o age group (p=0.811) and decreased slightly in patients in older age groups, especially in the 2 to 5 y/o age group (p=0.030). Conclusion: the pathology of thymus development in patients with DDS leads to impaired maturation of T-lymphocytes, leading to an increase in the number of mature forms of T-lymphocytes, shifts towards the development of T-helper 1 and T-helper 17, and a decrease in T-regulatory cells. Disturbances in T-lymphocytes can cause changes (dysregulation) in subpopulations of B-cells. All these processes may underlie the progression of autoimmune and infectious complications in such patients that are increasing with aging.

Comparison of Relative Activity versus Relative Expression Factors (RAF versus REF) in Predicting Glucuronidation Mediated Drug Clearance Using Recombinant UGTs.

Predicting the quantitative fraction of glucuronidation (fgluc) by individual UDP-glucuronosyltransferase enzymes (UGTs) is challenging due to the lack of selective inhibitors and inconsistent activity of recombinant UGT systems (rUGTs). Our study compares the relative expression versus activity factors (REF versus RAF) to predict fgluc based on rUGT data to human liver and intestinal microsomes (HLM and HIM). REF scalars were derived from a previous in-house proteomics study for eleven UGT enzymes (UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A9, UGT1A10, UGT2B4, UGT2B7, UGT2B10, UGT2B15, and UGT2B17), whereas RAF was calculated by measuring activities in rUGTs to microsomes of selective UGT probe substrates. Protein-normalized activity factor (pnAF) values weregenerated after correcting activity of individual UGTs to their corresponding protein abundance. The utility of REF and RAF in predicting fgluc was assessed for three UGT substrates-diclofenac, vorinostat, and raltegravir. The REF values ranged from 0.02 to 1.75, RAF based on activity obtained in rUGTs to HLM/HIM were from 0.1 to 274. pnAF values were ~ 5 to 80-fold, except for UGT2B4 and UGT2B15, where pnAF was ~ 180 and > 1000, respectively. The results revealed confounding effect of differential specific activities (per pmol) of rUGTs in fgluc prediction. The data suggest that the activity of UGT enzymes was significantly lower when compared to their activity in microsomes at the same absolute protein amount (pmol). Collectively, results of this study demonstrate poor and variable specific activity of different rUGTs (per pmol protein), as determined by pnAF values, which should be considered in fgluc scaling.

Donor Fractions of Cell-Free DNA Are Elevated During CLAD But Not During Infectious Complications After Lung Transplantation.

During the last few years, cell-free DNA (cfDNA) has emerged as a possible non-invasive biomarker for prediction of complications after lung transplantation. We previously published a proof-of-concept study using a digital droplet polymerase chain reaction (ddPCR)-based method for detection of cfDNA. In the current study, we aimed to further evaluate the potential clinical usefulness of detecting chronic lung allograft dysfunction (CLAD) using three different ddPCR applications measuring and calculating the donor fraction (DF) of cfDNA as well as one method using the absolute amount of donor-derived cfDNA. We analyzed 246 serum samples collected from 26 lung transplant recipients. Nine of the patients had ongoing CLAD at some point during follow-up. All four methods showed statistically significant elevation of the measured variable in the CLAD samples compared to the non-CLAD samples. The results support the use of ddPCR-detected cfDNA as a potential biomarker for prediction of CLAD. These findings need to be validated in a subsequent prospective study.

Liquid biopsy in brain tumors: moving on, slowly.

Due to limited access to the tumor, there is an obvious clinical potential for liquid biopsy in patients with primary brain tumors. Here, we review current approaches, present limitations to be dealt with, and new promising data that may impact the field. The value of circulating tumor cell-free DNA (ctDNA) in the cerebrospinal fluid (CSF) for the noninvasive diagnosis of primary brain tumors has been confirmed in several reports. The detection of ctDNA in the peripheral blood is desirable for patient follow-up but requires ultrasensitive methods to identify low mutant allelic frequencies. Digital PCR approaches and targeted gene panels have been used to identify recurrent hotspot mutations and copy number variations (CNVs) from CSF or plasma. Tumor classification from circulating methylomes in plasma has been actively pursued, although the need of advanced bioinformatics currently hampers clinical application. The use of focused ultrasounds to open the blood-brain barrier may represent a way to enrich of ctDNA the peripheral blood and enhance plasma-based liquid biopsy. Monitoring CNVs and hotspot mutations by liquid biopsy is a promising tool to detect minimal residual disease and strengthen response assessment in patients with primary brain tumors. Novel methods to increase the relative and/or absolute amount of ctDNA can improve the clinical potential of plasma-based liquid biopsies.

Fluorescence Lifetime Measurement For Studying Evaporation Of Bi-Component Droplet

Sprays are essential in engineering applications like evaporators and combustion chambers. Predicting the evaporation of multi-component liquid droplets is complex due to changing composition and properties. Measuring droplet composition is challenging, with limited non-intrusive methods available. Raman spectroscopy faces hurdles like weak signals and interference. Intensity based Laser-Induced Fluorescence (LIF) requires optical models to interpret data due to varying droplet size and position. Fluorescent dyes in liquid mixtures are sensitive to temperature, complicating measurements. Fluorescence lifetime measurements offer a promising alternative, being an absolute quantity less affected by reabsorption. This study develops a novel method using fluorescence lifetime to analyze droplet composition, leveraging Time-Correlated Single Photon Counting (TCSPC) for precise measurements. Factors such as dye concentration, solvent polarity, and temperature sensitivity are investigated. Calibration curves for ethanol-water mixtures correlate fluorescence lifetime with volume fraction. Experimental setups use TCSPC and femtosecond lasers for dye excitation. Results show that fluorescence lifetime decreases linearly with solvent polarity and is influenced by temperature and dye concentration. High dye concentrations cause self-quenching, altering temperature sensitivity. The method was applied to study the evaporation of ethanol-water droplets under acoustic levitation. Findings indicate that evaporation rates slow as the water fraction increases, with initial measurements showing higher water content due to partial evaporation. This research introduces a novel technique for droplet compositional analysis using fluorescence lifetime, providing insights into optimizing measurement accuracy and advancing understanding in engineering and related fields.