Each coin has 2 sides: a positive role of alien Potamopyrgus antipodarum (Grey, 1843) snails in reducing the infection of native lymnaeids with trematodes.

The change in the distribution of organisms in freshwater ecosystems due to natural or manmade processes raises the question of the impact of alien species on local communities. Although most studies indicate a negative effect, the positive one is more difficult to discern, especially in multispecies systems, including hosts and parasites. The purpose of the study was to check whether the presence of an alien host, Potamopyrgus antipodarum, reduces the intensity of Echinoparyphium aconiatum metacercariae in a native host, Radix spp. We additionally tested the impact of water temperature and the biomass of the alien host on the dilution effect. We experimentally studied (1) the lifespan of echinostome cercariae in different temperatures, (2) the infectivity of cercariae toward the alien host and native host, and (3) the impact of different biomass of the alien host on the intensity of metacercariae in the native host. We found that cercarial survival and infectivity were temperature dependent. However, cercarial survival decreased with increasing temperature, contrary to cercarial infectivity. Echinostome cercariae entered the renal cavity of both the native host and alien host, and successfully transformed into metacercariae. The number of metacercariae in the native host decreased with the increasing biomass of the alien host. Our results indicate that lymnaeids may benefit from the co-occurrence with P. antipodarum, as the presence of additional hosts of different origins may reduce the prevalence of parasites in native communities. However, the scale of the dilution effect depends not only on the increased spectrum of susceptible hosts but also on the other variables of the environment, including water temperature and host density.

Role of Bacillus subtilis exopolymeric genes in modulating rhizosphere microbiome assembly.

BACKGROUND: Bacillus subtilis is well known for promoting plant growth and reducing abiotic and biotic stresses. Mutant gene-defective models can be created to understand important traits associated with rhizosphere fitness. This study aimed to analyze the role of exopolymeric genes in modulating tomato rhizosphere microbiome assembly under a gradient of soil microbiome diversities using the B. subtilis wild-type strain UD1022 and its corresponding mutant strain UD1022eps-TasA, which is defective in exopolysaccharide (EPS) and TasA protein production. RESULTS: qPCR revealed that the B. subtilis UD1022eps-TasA- strain has a diminished capacity to colonize tomato roots in soils with diluted microbial diversity. The analysis of bacterial ß-diversity revealed significant differences in bacterial and fungal community structures following inoculation with either the wild-type or mutant B. subtilis strains. The Verrucomicrobiota, Patescibacteria, and Nitrospirota phyla were more enriched with the wild-type strain inoculation than with the mutant inoculation. Co-occurrence analysis revealed that when the mutant was inoculated in tomato, the rhizosphere microbial community exhibited a lower level of modularity, fewer nodes, and fewer communities compared to communities inoculated with wild-type B. subtilis. CONCLUSION: This study advances our understanding of the EPS and TasA genes, which are not only important for root colonization but also play a significant role in shaping rhizosphere microbiome assembly. Future research should concentrate on specific microbiome genetic traits and their implications for rhizosphere colonization, coupled with rhizosphere microbiome modulation. These efforts will be crucial for optimizing PGPR-based approaches in agriculture.

Probing cell proliferation: Considerations for dye selection.

Broadly speaking, cell tracking dyes are fluorescent compounds that bind stably to components on or within the cells so the fate of the labeled cells can be followed. Their staining should be bright and homogeneous without affecting cell function. For purposes of monitoring cell proliferation, each time a cell divides the intensity of cell tracking dye should diminish equally between daughter cells. These dyes can be grouped into two different classes. Protein reactive dyes label cells by reacting covalently but non-selectively with intracellular proteins. Carboxyfluorescein diacetate succinimidyl ester (CFSE) is the prototypic general protein label. Membrane intercalating dyes label cells by partitioning non-selectively and non-covalently within the plasma membrane. The PKH membrane dyes are examples of lipophilic compounds whose chemistry allows for their retention within biological membranes without affecting cellular growth, viability, or proliferation when used properly. Here we provide considerations based for labeling cell lines and peripheral blood mononuclear cells using both classes of dyes. Examples from optimization experiments are presented along with critical aspects of the staining procedures to help mitigate common risks. Of note, we present data where a logarithmically growing cell line is labeled with both a protein dye and a membrane tracking dye to compare dye loss rates over 6days. We found that dual stained cells paralleled dye loss of the corresponding single stained cells. The decrease in fluorescence intensity by protein reactive dyes, however, was more rapid than that with the membrane reactive dyes, indicating the presence of additional division-independent dye loss.

Impact of nutritional factors on the in vitro PK/PD modelling of polymyxin B against various strains of Acinetobacter baumannii.

The main objective of this study was to assess the effect of rich artificial cation adjusted Mueller-Hinton broth (CAMHB), on the growth of three Acinetobacter baumannii strains, ATCC 19606 and two clinical A. baumannii strains, either susceptible or resistant to polymyxin B (PMB), and on the PMB bactericidal activity. A pharmacokinetic (PK) / pharmacodynamic (PD) modelling approach was then used to characterize the effect of PMB in various conditions. Time-kill experiments were performed using undiluted CAMHB or diluted at 50%, 25% and 10%, with or without Ca2+ and Mg2+ compensation (known to affect PMB activity), and with PMB concentrations ranging from 0.25 mg/L to 256 mg/L based on the strain's MIC. For each strain, time-kill replicates were modelled using NONMEM®. Unexpectedly, CAMHB dilution by up to 10-fold did not affect the growth rate of any of the three strains in the absence of PMB. Yet, PMB bactericidal activity increased with medium dilution resulting in particular in a reduction of the apparent bacterial regrowth of the various strains observed typically after few hours of experiment. The data of each strain were well characterised by a PK/PD model with two bacterial sub-populations with different susceptibility to PMB (a more susceptible (S+) and a less susceptible (S-)). Relatively large, unexplained strain-to-strain variability was observed regarding the impact of medium dilution as well as cation compensation. Complementary experiments are needed to characterise the mechanism underlying this medium dilution effect.

Formation of Key Aroma Compounds During 30 Weeks of Ripening in Gouda-Type Cheese Produced from Pasteurized and Raw Milk.

Gouda-type cheeses were produced on a pilot-scale from raw milk (RM-G) and pasteurized milk (PM-G). Sixteen key aroma compounds previously characterized by the sensomics approach were quantitated in the unripened cheeses and at five different ripening stages (4, 7, 11, 19, and 30 weeks) by means of stable isotope dilution assays. Different trends were observed in the formation of the key aroma compounds. Short-chain free fatty acids and ethyl butanoate as well as ethyl hexanoate continuously increased during ripening but to a greater extent in RM-G. Branched-chain fatty acids such as 3-methylbutanoic acid were also continuously formed and reached a 60-fold concentration after 30 weeks, in particular in PM-G. 3-Methylbutanal and butane-2,3-dione reached a maximum concentration after 7 weeks and decreased with longer ripening. Lactones were high in the unripened cheeses and increased only slightly during ripening. Recent results have shown that free amino acids were released during ripening. The aroma compounds 3-methylbutanal, 3-methyl-1-butanol, and 3-methylbutanoic acid are suggested to be formed by microbial enzymes degrading the amino acid l-leucine following the Ehrlich pathway. To gain insight into the quantitative formation of each of the three aroma compounds, the conversion of the labeled precursors (13C6)-l-leucine and (2H3)-2-keto-4-methylpentanoic acid into the isotopically labeled aroma compounds was studied. By applying the CAMOLA approach (defined mixture of labeled and unlabeled precursor), l-leucine was confirmed as the only precursor of the three aroma compounds in the cheese with the preferential formation of 3-methylbutanoic acid.

Influence of Milk Pasteurization on the Key Aroma Compounds in a 30 Weeks Ripened Pilot-Scale Gouda Cheese Elucidated by the Sensomics Approach.

Gouda cheese was produced from pasteurized milk and ripened for 30 weeks (PM-G). By application of gas chromatography/olfactometry and an aroma extract dilution analysis on the volatiles isolated by extraction/SAFE distillation, 25 odor-active compounds in the flavor dilution (FD) factor range from 16 to 4096 were identified. Butanoic acid, 2- and 3-methylbutanoic acid, and acetic acid showed the highest FD factors, and 2-phenylethanol, δ-decalactone, and δ-dodecalactone were most odor-active in the neutral-basic fraction. Quantitations by stable isotope dilution assays followed by a calculation of odor activity values (OAVs) revealed acetic acid, 3-methylbutanoic acid, butanoic acid, and butane-2,3-dione with the highest OAVs. Finally, an aroma recombinate prepared based on the quantitative data well agreed with the aroma profile of the PM-G. In Gouda cheese produced from raw (nonpasteurized) milk (RM-G), qualitatively the same set of odor-active compounds was identified. However, higher OAVs of butanoic acid, hexanoic acid, and their corresponding ethyl esters were found. On the other hand, in the PM-G, higher OAVs for 3-methylbutanoic acid, 3-methylbutanol, 3-methylbutanal, and butane-2,3-dione were determined. The different rankings of these key aroma compounds clearly reflect the aroma differences of the two Gouda-type cheeses. A higher activity of lipase in the RM-G and higher amounts of free l-leucine in PM-G on the other side were responsible for the differences in the concentrations of some key aroma compounds.

The effect of diversity on disease reverses from dilution to amplification in a 22-year biodiversity × N × CO2 experiment.

Plant disease often increases with N, decreases with CO2, and increases as biodiversity is lost (i.e., the dilution effect). Additionally, all these factors can indirectly alter disease by changing host biomass and hence density-dependent disease transmission. Yet over long periods of time as communities undergo compositional changes, these biomass-mediated pathways might fade, intensify, or even reverse in direction. Using a field experiment that has manipulated N, CO2, and species richness for over 20 years, we compared severity of a specialist rust fungus (Puccinia andropogonis) on its grass host (Andropogon gerardii) shortly after the experiment began (1999) and twenty years later (2019). Between these two sampling periods, two decades apart, we found that disease severity consistently increased with N and decreased with CO2. However, the relationship between diversity and disease reversed from a dilution effect in 1999 (more severe disease in monocultures) to an amplification effect in 2019 (more severe disease in mixtures). The best explanation for this reversal centered on host density (i.e., aboveground biomass), which was initially highest in monoculture, but became highest in mixtures two decades later. Thus, the diversity-disease pattern reversed, but disease consistently increased with host biomass. These results highlight the consistency of N and CO2 as drivers of plant disease in the Anthropocene and emphasize the critical role of host biomass-despite potentially variable effects of diversity-for relationships between biodiversity and disease.

Developing Salt-Rejecting Evaporators for Solar Desalination: A Critical Review.

Solar desalination, a green, low-cost, and sustainable technology, offers a promising way to get clean water from seawater without relying on electricity and complex infrastructures. However, the main challenge faced in solar desalination is salt accumulation, either on the surface of or inside the solar evaporator, which can impair solar-to-vapor efficiency and even lead to the failure of the evaporator itself. While many ideas have been tried to address this ″salt accumulation″, scientists have not had a clear system for understanding what works best for the enhancement of salt-rejecting ability. Therein, for the first time, we classified the state-of-the-art salt-rejecting designs into isolation strategy (isolating the solar evaporator from brine), dilution strategy (diluting the concentrated brine), and crystallization strategy (regulating the crystallization site into a tiny area). Through the specific equations presented, we have identified key parameters for each strategy and highlighted the corresponding improvements in the solar desalination performance. This Review provides a semiquantitative perspective on salt-rejecting designs and critical parameters for enhancing the salt-rejecting ability of dilution-based, isolation-based, and crystallization-based solar evaporators. Ultimately, this knowledge can help us create reliable solar desalination solutions to provide clean water from even the saltiest sources.

Comparison of the Mechanical Properties of Hardfacings Made by Standard Coated Stick Electrodes and a Newly Developed Rectangular Stick Electrode.

Cladding with a stick electrode is one of the oldest arc processes for adding a deposit on a base material. The process is suitable for outdoor working, but the disadvantages are low productivity and large dilution rates. In this work, a simple solution is proposed, which would enable cladding of a larger area with one pass and decrease the dilution rate at the same time-a new type of electrode was developed, exhibiting a rectangular cross-section instead of a round one. Hardfacings, welded with E Fe8 electrodes according to EN 14 700 Standard were welded on mild steel S355 J2 base material with three different coated stick electrodes. The first one was a commercially available, standard, round hardfacing electrode, the second was the same, but with a thinner coating, and the third one was a newly developed rectangular electrode. All three types had equal cross-sections of the metallic core and the same type of coating. Manufacturing of the rectangular electrodes in the laboratory is explained briefly. One- and multi-layer deposits were welded with all three types. Differences were observed in the arc behavior between the round and rectangular electrodes. With the rectangular electrode, the microstructure of the deposit was finer, penetration was shallower, and dilution rates were lower, while the hardness was higher, residual stresses predominantly compressive, and the results of instrumented Charpy impact tests and fracture mechanics tests were better.

Key Odorants Forming Aroma of Polish Mead: Influence of the Raw Material and Manufacturing Processes.

Mead was analyzed by using the concept of molecular sensory science for the identification of key odorants. A total of 29 odor-active compounds were identified in mead by using gas chromatography olfactometry (GCO). Flavor dilution (FD) factors of identified compounds ranged from 1 to 16,384, compounds with FD factors ≥32 were quantitated by using stable isotopically substituted odorants as internal standards or external standard method, and odor activity values (OAVs) were calculated. Fifteen compounds showed OAVs ≥1: aldehydes (2-phenylacetaldehyde, 3-(methylsulfanyl)propanal), 4-hydroxy-3-methoxybenzaldehyde), esters (ethyl 3-methylbutanoate, ethyl propanoate, ethyl octanoate), alcohols (2-phenylethan-1-ol, 3- and 2-methylbutan-1-ol, 3-(methylsulyfanyl)propan-1-ol), furanons (4-hydroxy-2,5-dimethylfuran-3(2H)-one, 3-hydroxy-4,5-dimethylfuran-2(5H)-one), acids (3- and 2-methylbutanoic acid, acetic acid), 1,1-diethoxyethane, and 4-methylphenol. 2-Phenylacetaldehyde (OAV, 3100) was suggested as the compound with the biggest influence on the aroma of mead, followed by 4-hydroxy-2,5-dimethylfuran-3(2H)-one (OAV, 1900), 3-(methylsulfanyl)propanal (OAV, 890), and 2-phenylethan-1-ol (OAV, 680). Quantitative olfactory profile analysis revealed strong honey, malty, and alcoholic impressions. Omission experiments revealed that 3-(methylsulfanyl)propanal, 2-phenylethan-1-ol, 4-hydroxy-2,5-dimethylfuran-3(2H)-one, ethyl propanoate, ethyl 3-methylbutanoate, 2-phenylacetaldehyde, 3- and 2-methylbutanoic acid, 3-hydroxy-4,5-dimethylfuran-2(5H)-one, and 4-hydroxy-3-methoxybenzaldehyde were the key odorants in the mead. Determining concentrations of key odorants in important production steps showed that the fermentation and maturation stages had the strongest effect on the formation of mead aroma.

The gamma-variate in contrast-enhanced imaging: a unified view and method from computed to electrical impedance tomography.

OBJECTIVE: Modern medical imaging plays a vital role in clinical practice, enabling non-invasive visualization of anatomical structures. Dynamic contrast enhancement (DCE) imaging is a technique that uses contrast agents to visualize blood flow dynamics in a time-resolved manner. It can be applied to different modalities, such as computed tomography (CT) and electrical impedance tomography (EIT). This study aims to develop a common theoretical and practical hemodynamic extraction basis for DCE modelling across modalities, based on the gamma-variate function. Approach: The study introduces a framework to generate time-intensity curves for multiple DCE imaging modalities from user-defined hemodynamic parameters. Thus, extensive datasets were simulated for both DCE-CT and EIT, representing different hemodynamic scenarios. Additionally, gamma-variate extensions to account for several physiological effects were detailed in a modality-agnostic manner, and three corresponding fitting strategies, namely nonlinear, linear, and a novel hybrid approach, were implemented and compared on the basis of accuracy of parameter estimation, first pass reconstruction, speed of computation, and failure rate. Main results: As a result, we found the linear method to be the most modality-dependent, exhibiting the greatest bias, variance and failure rates, although remaining the fastest alternative. The hybrid method at least matches the state-of-the-art nonlinear method's accuracy, while improving its robustness and speed by 10 times. Significance: Our research suggests that the hybrid method may bring noteworthy accuracy and efficiency improvements in handling the high-dimensionality of DCE imaging in general, being a step towards real-time processing. Moreover, our generative model presents a potential asset to produce benchmarking and data augmentation datasets across modalities.

In-vitro antioxidant, antimicrobial and phytochemical properties of extracts from the pulp and seeds of the African baobab fruit (Adansonia digitata L.).

Adansonia digitata, commonly known as the African Baobab plant is used widely in traditional medicine for treating of many diseases. The current study investigates the antioxidant and antimicrobial properties, and nutritional composition of the pulp and seeds from the fruit of African Baobab plant. Matured fruits were harvested and processed by separating the fruit pulp and seeds. Water, 70 % Ethanol/water mixture, and Hexane were used as solvents for extraction. Antioxidant properties of extracts in this study were investigated using the 2,2-diphenyl-1-picrylhydrazyl, hydrogen peroxidescavenging assays., Total Flavonoid Content, Total Phenolic Content, Total Tannin Content, and Total Antioxidant Capacity were also investigated. Agar Well Diffusion and Broth Dilution methods were used to estimate the antimicrobial properties of the extracts. The proximate composition of the seeds and fruit pulps was also determined. GC-MS was employed to determine the fatty acid composition. Results obtained showed the presence of Total phenolics (range 4.1-5.5 mg GAE/g), Total flavonoids (range 10.1-16.5 mg QE/g), Total Tannins (range 1.7-15.6 mg CE/g), and Antioxidants (range 2.0-14.5 mg AAE/g). The H2O2 and DPPH assays gave IC50s in the ranges of 300-1800 mg/L and 700-1600 mg/L respectively. Extract from the fruit pulp was found to inhibit the growth of a panel of 2 g-positive bacteria, 2 g-negative bacteria, and two fungi microorganisms. Fatty acids such as myristic acid, palmitic acid, and stearic acid were found to be present in oil from the seeds. Proximate components such as crude protein, crude fat, and crude fibre were found to be high. From the results, seeds and the fruit pulp of the African Baobab plant have significant antioxidant properties and can inhibit microbial growth.

The dilution evaluation as a corrective measure for interference in the white blood cell scattergram in Beckman Coulter DxH 900.

BACKGROUND: The Beckman Coulter DxH 900 is a haematological analyser capable of counting and sizing blood cells, and obtaining a complete blood cell count (CBC). This analyses different parameters of red blood cells (RBC), platelets and white blood cells/leukocytes. Some automated CBC counters present limitations due to specimen characteristics, abnormal cells or both factors. In the presence of abnormalities, the DxH 900 has a flagging system, warning the laboratory technician that something needs to be verified. In the present work, we evaluated samples from oncologic patients, presenting a population erroneously perceived as being lymphocytes. The most common explanations for this situation are RBC resistant to lysis or serum hyperbilirubinaemia. METHODS: In an attempt to solve and understand what the cause of this problem might be, we diluted our samples (1:3) and analysed the serum total bilirubin. To identify cells' abnormalities, the samples were also analysed by manual DLC counts. During the study, we also checked the different flags presented by the equipment. RESULTS: The results evidenced that the major interference was due to RBC lysis resistance, corresponding to 94.7% of the cases, while hyperbilirubinaemia was only present in 73.4%. Besides, we determined that some samples with normal bilirubin levels also presented interference, suggesting that hyperbilirubinaemia was not the main cause of the error. The most recurrent flag observed was "High event rate". CONCLUSION: The dilution solved all of the observed interferences. The results between diluted and manual counts showed a strong correlation, leading us to introduce dilution in our laboratory routine.

Methods for screening and evaluation of antimicrobial activity: A review of protocols, advantages, and limitations.

Infectious diseases pose a formidable global challenge, compounded by the emergence of antimicrobial resistance. Consequently, researchers are actively exploring novel antimicrobial compounds as potential solutions. This endeavor underscores the pivotal role of methods employed for screening and evaluating antimicrobial activity-a critical step in discovery and characterization of antimicrobial agents. While traditional techniques such as well-diffusion, disk-diffusion, and broth-dilution are commonly utilized in antimicrobial assays, they may encounter limitations concerning reproducibility and speed. Additionally, a diverse array of antimicrobial assays including cross-streaking, poisoned-food, co-culture, time-kill kinetics, resazurin assay, bioautography, etc., are routinely employed in antimicrobial evaluations. Advanced techniques such as flow-cytometry, impedance analysis, and bioluminescent technique may offer rapid and sensitive results, providing deeper insights into the impact of antimicrobials on cellular integrity. However, their higher cost and limited accessibility in certain laboratory settings may present challenges. This article provides a comprehensive overview of assays designed to characterize antimicrobial activity, elucidating their underlying principles, protocols, advantages, and limitations. The primary objective is to enhance understanding of the methodologies designed for evaluating antimicrobial agents in our relentless battle against infectious diseases. By selecting the appropriate antimicrobial testing method, researchers can discern suitable conditions and streamline the identification of effective antimicrobial agents.

Pulmonary Transit Time Assessment by CEUS in Healthy Rabbits: Feasibility, and the Effects of UCAs Dilution Concentration.

To evaluate the inter-observer variability and the intra-observer repeatability of pulmonary transit time (PTT) measurement using contrast-enhanced ultrasound (CEUS) in healthy rabbits, and assess the effects of dilution concentration of ultrasound contrast agents (UCAs) on PTT. Thirteen healthy rabbits were selected, and five concentrations UCAs of 1:200, 1:100, 1:50, 1:10, and 1:1 were injected into the right ear vein. Five digital loops were obtained from the apical 4-chamber view. Four sonographers obtained PTT by plotting the TIC of right atrium (RA) and left atrium (LA) at two time points (T1 and T2). The frame counts of the first appearance of UCAs in RA and LA had excellent inter-observer agreement, with intra-class correlations (ICC) of 0.996, 0.988, respectively. The agreement of PTT among four observers was all good at five different concentrations, with an ICC of 0.758-0.873. The reproducibility of PTT obtained by four observers at T1 and T2 was performed well, with ICC of 0.888-0.961. The median inter-observer variability across 13 rabbits was 6.5% and the median variability within 14 days for 4 observers was 1.9%, 1.7%, 2.2%, 1.9%, respectively; The PTT of 13 healthy rabbits is 1.01 ± 0.18 second. The difference of PTT between five concentrations is statistically significant. The PTT obtained by a concentration of 1:200 and 1:100 were higher than that of 1:1, while there were no significantly differences in PTT of a concentration of 1:1, 1:10, and 1:50. PTT measured by CEUS in rabbits is feasible, with excellent inter-observer and intra-observer reliability and reproducibility, and dilution concentration of UCAs influences PTT results.

Development of an SI-traceable N-terminal pro-B-type natriuretic peptide certified reference material using structure-based impurity-corrected isotope dilution mass spectrometry approaches.

N-Terminal pro-B-type natriuretic peptide (NT-proBNP) is a pivotal biomarker for the diagnosis and prognosis of heart failure (HF). However, no SI-traceable certified reference material (CRM) or reference measurement procedure (RMP) is available for this biomarker, and so clinical testing results obtained in different laboratories cannot be traced to a higher-order standard, leading to incomparable measurements. Protein hydrolysis and protein cleavage isotope dilution mass spectrometry (AAA-IDMS and PepA-IDMS) were used to develop a CRM. Structurally related impurities were identified by high-resolution mass spectrometry. The quantitative AAA-IDMS results were corrected according to the amino acid compositions of the impurities. Using PepA-IDMS, two peptides from the proteolyzed product were confirmed as signature peptides. To obtain traceable and accurate results, the signature peptides were quantified using impurity-corrected AAA-IDMS. The candidate NT-proBNP solution was denatured and enzymatically digested using the Glu-C endoproteinase. The released signature peptides were measured using an isotopic dilution approach. The homogeneity and stability of the candidate CRM were characterized, and their uncertainties were combined with the value assignment process. The developed CRM can be considered a unique SI-traceable NT-proBNP reference material and is expected to be used as a primary calibrator for matrix NT-proBNP CRM development.

A nonionic microemulsion co-loaded with atorvastatin and quercetin: Simultaneous spectroscopic analysis and payload release kinetics.

In this study, we have co-loadedatorvastatin (ATR) and quercetin (QCT) in a nonionic microemulsion. After developing a derivative ratio spectrophotometric technique for simultaneous analysis of ATR and QCT, pseudoternary phase diagram was constructed utilizing1:4 d-α-tocopherol polyethylene glycol 1000 succinate (TPGS) and ethanol as surfactant and cosurfactant, respectively. Oleic acid was used as oil phase. Structural characterization of the formulation was carried out along a water dilution line created in monophasic region. Characterizations at these dilution points were performed using dynamic light scattering and polarized light microscopy. The average hydrodynamic size of the optimized formulation was found to be 18.9 nm and it did not change upon loading of ATR and QCT. In vitro release was assessed for the formulations loaded with different ratios of ATR and QCT, and the data were fitted to different mathematical models. Interestingly, we noticed differences in release kinetics during changes in dose ratios, particularly for QCT. Higuchi kinetics, observed at equal dose, shifted to Korsmeyer-Peppas model at higher QCT-ATR ratio (2:1 and 4:1). This difference is attributable to the ability of QCT molecules of overwhelming the interface at higher concentrations. Altogether, our observations highlight that the ratio of payloads should be selected carefully in order to avoid unpredictable release patterns.

Bioaccumulation and trophic transfer of per- and polyfluoroalkyl substances in a subtropical mangrove estuary food web.

Mangrove estuaries are an important land-sea transitional ecosystem that is currently under various pollution pressures, while there is a lack of research on per- and polyfluoroalkyl substances (PFAS) in the organisms of mangrove estuaries. In this study, we investigated the distribution and seasonal variation of PFAS in the tissues of organisms from a mangrove estuary. The PFAS concentrations in fish tissues varied from 0.45 ng/g ww to 17.67 ng/g ww and followed the order of viscera > head > carcass > muscle, with the highest tissue burden found in the fish carcass (39.59 ng). The log BAF values of PFDoDA, PFUnDA, and PFDA in the whole fish exceeded 3.70, indicating significant bioaccumulation. The trophic transfer of PFAS in the mangrove estuary food web showed a dilution effect, which was mainly influenced by the spatial heterogeneity of PFAS distribution in the estuarine environment, and demonstrated that the gradient dilution of PFAS in the estuary habitat environment can disguise the PFAS bio-magnification in estuarine organisms, and the larger the swimming ranges of organisms, the more pronounced the bio-dilution effect. The PFOA-equivalent HRs of category A and B fish were 3.48-5.17 and 2.59-4.01, respectively, indicating that mangrove estuarine residents had a high PFAS exposure risk through the intake of estuarine fish.

Automated ELISA for potency measurements of therapeutic antibodies and antibody fragments.

Potency assays are essential for the development and quality control of biopharmaceutical drugs, but they are often a time limiting factor due to manual handling steps and consequently low analytical throughput. On the other hand, automation of potency assays can be challenging due to their complexity and the use of biological materials. ELISA (enzyme-linked immunosorbent assay) is widely used for potency determination and is a good candidate for automation as all ELISA types depend on the same basic steps: coating, blocking, sample incubation, detection, and signal measurement. Nevertheless, ELISA for relative potency measurements still require drug-specific development and assay validation thereby complicating automation efforts. To simplify potency testing by ELISA, we first developed a manual protocol generally applicable to different drugs and then adapted this protocol for automated measurements. We identified unexpected critical parameters which had to be adapted to transfer the manual ELISA to an automated liquid handling system and we demonstrated that gravimetric sample dilution is unnecessary with the automated protocol. Both manual and automated protocols were validated and compared using multiple biotherapeutics. The automated protocol showed similar or higher precision and accuracy when compared to the manual method.

Comparison of series and parallel reactance to identify changes in intracellular water in response to physical training in athletes during a sports season.

OBJECTIVE: Cross-sectional evidence has demonstrated that parallel reactance obtained by bioelectrical impedance analysis (BIA) may be an alternative to the regularly used series of measurements to predict intracellular water (ICW) in athletes. However, we are not aware of any studies that have determined the predictive role or compared the effectiveness of both series and parallel reactance for tracking ICW changes during an athletic season. The main aim of this study was to determine the predictive role and compare both series and parallel reactance (Xc) in tracking ICW during an athletic season. RESEARCH METHODS AND PROCEDURES: This longitudinal study analyzed 108 athletes in the preparatory and competitive periods. Using dilution techniques, total body water (TBW) and extracellular water (ECW) were determined and ICW was calculated. Resistance (R), Xc, and impedance (Z) standardized for height were obtained through BIA spectroscopy using a frequency of 50kHz in a series array and then mathematically transformed in a parallel array. RESULTS: Multiple regression analyses showed that only changes in parallel Xc and capacitance (CAP) (P < 0.05) were predictors of delta ICW during the sports season. In contracts, this was not the case for Xcs. Both changes in R and Z, series and parallel, predicted similarly the changes in ECW and TBW (P < 0.05) in athletes. CONCLUSION: Our findings highlight the potential of parallel BIA values to detect changes in body water compartments over a competitive season. These data provide preliminary evidence that changes in parallel Xc/H, and ultimately CAP, represent valid markers of alterations in cell volume during a sports season.