Uptake Phase Research Articles

Rapid screening of shellfish tainting from oil spills using an antibody-based biosensor.

Tainting of shellfish by polyaromatic hydrocarbons (PAHs) following an oil spill poses possible health risks as well as socioeconomic impacts. Traditional screening approaches for evaluating PAH contamination have limitations that can prevent timely, objective spill response decisions. The objective of this study was to investigate the relationship between PAH concentrations measured in the oyster, Crassostrea virginica, interstitial fluid using a rapid antibody-based biosensor method, with PAH concentrations in oyster tissues determined using conventional gas chromatography-mass spectrometry analysis. To accomplish this objective, bioconcentration tests were performed to simulate oil spill exposures using a crude and heavy fuel oil containing different PAH compositions. This design allowed both the PAH concentration and composition in water and, subsequently, accumulated by oysters to be varied over time. Oysters sampled during uptake and depuration phases were analyzed using biosensor and conventional analysis methods to generate comparative data. Results indicated that biosensor measurements of oysters captured the kinetics of PAH accumulation during uptake and depuration phases. Further, significant positive correlations were observed between biosensor interstitial fluid and lipid-normalized PAH tissue concentrations. However, quantitative predictions appear to be modulated by the contamination source and target analyte list for tissue analysis. Thus, the biosensor can be applied for rapidly evaluating relative PAH contamination between biota samples and offers a promising new analytical tool for oil spill monitoring and fisheries management contexts. A generic model was also developed from study and literature data to predict PAH half-lives from bivalve tissues. These predictions can help inform field monitoring of shellfish and estimate recovery times required to achieve pre-spill conditions.

Kinetics of methane hydrate formation in a 1200 ml unstirred reactor for natural gas storage

Kinetics of methane hydrate formation in a 1200 ml unstirred reactor for natural gas storage

Toxicokinetics of selenate in earthworm sub-tissues and potential bio-accessibility assessment of earthworm-derived selenium

Selenium (Se) pollution is mainly caused by anthropogenic activities, and the resulting biosecurity concerns have garnered significant attention in recent years. Using one-compartmental toxicokinetic (TK) modelling, this study explored the kinetic absorption, sub-tissue distribution, and elimination processes of the main Se species (selenate, Se(VI)) in the cultivated aerobic soil of the earthworm Eisenia fetida. The bio-accessibility of earthworm-derived Se was assessed using an in vitro simulated gastrointestinal digestion test to evaluate its potential trophic risk. The results demonstrated that Se accumulated in the pre-clitellum (PC) and total tissues (TT) of earthworms in a time- and dose-dependent manner. The highest Se levels in the PC, post-clitellum (PoC), and TT were 70.54, 57.93, and 64.26 mg/kg during the uptake phase, respectively. The kinetic Se contents in the earthworms PC and TT were consistent with the TK model but not with PoC. The earthworm TT exhibited a faster uptake (Kus = 0.83-1.02 mg/kg/day) and elimination rate of Se (Kee = 0.044-0.049 mg/kg/day), as well as a shorter half-life time (LT1/2 = 15.88-14.22 days) than PC at low soil Se levels (≤5 mg/kg). Conversely, the opposite trend was observed with higher Se concentrations (10 and 20 mg/kg). These results are likely attributable to the tissue specificity and concentration of the toxicant. Earthworms PC and TT exhibited a higher kinetic Se accumulation factor (BAFk) than steady-state BAF (BAFss), with values ranging from 8 to 24 and 3-13, respectively. Furthermore, the bio-accessibility of earthworm-derived Se to poultry ranged from 66.25 % to 84.35 %. As earthworms are at the bottom of the terrestrial food chain, the high bio-accessibility of earthworm-derived Se poses a potential risk to predators. This study offers data support and a theoretical foundation for understanding the biological footprint of soil Se and its toxicological impacts and ecological hazards.

Enantioselective bioaccumulation, biotransformation and spatial distribution of chiral fungicide difenoconazole in earthworms (Eisenia fetida)

The enantioselective environmental behavior of difenoconazole, a widely utilized triazole fungicide commonly detected in agricultural soils, has yet to be comprehensively explored within the earthworm-soil system. To address this research gap, we investigated the bioaccumulation and elimination kinetics, degradation pathways, biotransformation mechanisms, spatial distribution, and toxicity of chiral difenoconazole. The four stereoisomers of difenoconazole were baseline separated and analyzed using SFC-MS/MS. Pronounced enantioselectivity was observed during the uptake phase, with earthworms exhibiting a preference for (2R,4R)-difenoconazole and (2R,4S)-difenoconazole. A total of five transformation products (TPs) were detected and identified using UHPLC-QTOF/MS in the earthworm-soil system. Four of the TPs were detected in both earthworm and soil, and one TP was produced only in eaerthwroms. Hydrolysis and hydroxylation were the primary transformation pathways of difenoconazole in both earthworms and soil. Furthermore, a chiral TP, 3-chloro, 4-hydroxy difenoconazole, was generated with significant enantioselectivity, and molecular docking results indicate the greater catalytic bioactivity of (2R,4R)- and (2R,4S)-difenoconazole, leading to the preferential formation of their corresponding hydroxylated TPs. Furthermore, Mass Spectrometry Imaging (MSI) was applied for the first time to explore the spatial distribution of difenoconazole and the TPs in earthworms, and the "secretory zone" was found to be the dominant region to uptake and biodegrade difenoconazole. ECOSAR predictions highlighted the potentially hazardous impact of most difenoconazole TPs on aquatic ecosystems. These findings are important for understanding the environmental fate of difenoconazole, evaluating environmental risks, and offering valuable insights for guiding scientific bioremediation efforts.

Pharmacokinetics of desflurane uptake and disposition in piglets.

Many respiratory but few arterial blood pharmacokinetics of desflurane uptake and disposition have been investigated. We explored the pharmacokinetic parameters in piglets by comparing inspiratory, end-tidal, arterial blood, and mixed venous blood concentrations of desflurane. Seven piglets were administered inspiratory 6% desflurane by inhalation over 2h, followed by a 2-h disposition phase. Inspiratory and end-tidal concentrations were detected using an infrared analyzer. Femoral arterial blood and pulmonary artery mixed venous blood were sampled to determine desflurane concentrations by gas chromatography at 1, 3, 5, 10, 20, 30, 40, 50, 60, 80, 100, and 120min during each uptake and disposition phase. Respiratory and hemodynamic parameters were measured simultaneously. Body uptake and disposition rates were calculated by multiplying the difference between the arterial and pulmonary artery blood concentrations by the cardiac output. The rates of desflurane body uptake increased considerably in the initial 5min (79.8ml.min-1) and then declined slowly until 120min (27.0ml.min-1). Similar characteristics of washout were noted during the subsequent disposition phase. Concentration-time curves of end-tidal, arterial, and pulmonary artery blood concentrations fitted well to zero-order input and first-order disposition kinetics. Arterial and pulmonary artery blood concentrations were best fitted using a two-compartment model. After 2h, only 21.9% of the desflurane administered had been eliminated from the body. Under a fixed inspiratory concentration, desflurane body uptake in piglets corresponded to constant zero-order infusion, and the 2-h disposition pattern followed first-order kinetics and best fitted to a two-compartment model.

Modeling Doxorubicin Pharmacokinetics in Multiple Myeloma Suggests Mechanism of Drug Resistance.

Multiple myeloma (MM) is a plasma cell malignancy often treated with chemotherapy drugs. Among these, doxorubicin (DOXO) is commonly employed, sometimes in combined-drug therapies, but it has to be optimally administered in order to maximize its efficacy and reduce possible side effects. To support DOXO studies and treatment optimization, here we propose an experimental/modeling approach to establish a model describing DOXO pharmacokinetics (PK) in MM cells. A series of in vitro experiments were performed in MM1R and MOLP-2 cells. DOXO was administered at two dosages (200 nM, 450 nM) at [Formula: see text] = 0 and removed at [Formula: see text] = 3 hrs. Intracellular DOXO concentration was measured via fluorescence microscopy during both drug uptake ([Formula: see text] = 0-3 hrs) and release phases ([Formula: see text] = 3-8 hrs). Four PK candidate models were identified, and were compared and selected based on their ability to describe DOXO data and numerical parameter identification. The most parsimonious model consists of three compartments describing DOXO distribution between the extracellular space, the cell cytoplasm and the nucleus, and defines the intracellular DOXO efflux rate through a Hill function, simulating a threshold/saturation drug resistance mechanism. This model predicted DOXO data well in all the experiments and provided precise parameter estimates (mean ± standard deviation coefficient of variation: 15.8 ± 12.2%). A reliable PK model describing DOXO uptake and release in MM cells has been successfully developed. The proposed PK model, once integrated with DOXO pharmacodynamics, has the potential of allowing the study and the optimization of DOXO treatment strategies in MM.

A first field evaluation of heavy metals uptake by innovative artificial mussel (AM) technology versus live mussel (LM) from inshore of the Bay of Bengal, Bangladesh and the application of AM technology for global pollution study

A first field evaluation of heavy metals uptake by innovative artificial mussel (AM) technology versus live mussel (LM) from inshore of the Bay of Bengal, Bangladesh and the application of AM technology for global pollution study

Tracking Nano- and Microplastics Accumulation and Egestion in a Marine Copepod by Novel Fluorescent AIEgens: Kinetic Modeling of the Rhythm Behavior.

Nano- and microplastics (NMPs) are now prevalent in the marine environment. This study quantified the uptake and depuration kinetics of spherical polystyrene NMPs of different particle sizes (200 nm/30 μm) and functional groups (-NH2/-COOH) in a temperate calanoid copepod Calanus sinicus (C. sinicus), which exhibited rhythmic feeding patterns in natural environments. Aggregated-induced emission (AIE) fluorescent probes were employed to track and quantify the kinetics of NMPs with excellent photostability and biocompatibility. The results showed that C. sinicus consumed all NMPs types, with preference of NMPs to small size and amino group. Increased diatom concentrations also inhibited the bioaccumulation of NMPs. Influenced by rhythmic behavior, the bioaccumulation of NMPs by C. sinicus was nonstationary during the 6 h uptake phase. After 1-3 h of rapid uptake, the body burden peaked and then slowly declined. During the 3 h depuration phase, C. sinicus rapidly and efficiently removed NMPs with a mean half-life of only 0.23 h. To further quantify the body burden of C. sinicus under the influence of rhythmic feeding behavior, a biokinetic model was established, and the Markov chain Monte Carlo method was used to estimate the parameter distribution. Our results highlighted that copepods exhibited unique rhythmic feeding behavior under environmentally relevant concentrations of NMPs exposure, which may influence the bioaccumulation, trophic transfer, and environmental fate of NMPs.

Lu-177-PSMA dosimetry for kidneys and tumors based on SPECT images at two imaging time points.

Personalized dosimetry for Lu-177-PSMA treatment requires multiple-time-point SPECT/CT scans to calculate time-integrated activity (TIA). This study evaluates two-time-point (TTP) methods for TIA calculation for kidneys and tumors. A total of 18 patients treated with 3.7-7.4 GBq Lu-177 PSMA-617 were analyzed retrospectively, including 18 sets of left and right kidneys, as well as 45 tumors. Four quantitative SPECT/CT (4TP) were acquired at 2 h, 20 h, 40 h, 60 h (n = 11), or 200 h (n = 7) after treatment, and they were fit bi-exponentially as reference. The TTP method was fitted by a mono-exponential washout function using two selected imaging time points for kidneys. For tumors, one uptake and one washout phase were modeled, assuming linear (type I) and same (type II) uptake phase between 0 h to the first time point and mono-exponential washout thereafter. Two single-time-point (STP) methods were also implemented for comparison. TIA calculated by TTP and STP methods were compared with reference to the 4TP TIA. For the kidneys, the TTP methods using 20 h-60 h and 40 h-200 h had smaller mean absolute errors of 8.05 ± 6.05% and 4.95 ± 3.98%, respectively, as compared to other combinations of time points and STP methods. For tumors, the type I and type II TTP methods using 20h-60 h and 40-200 h had smaller mean absolute errors of 6.14 ± 5.19% and 12.22 ± 4.44%, and 8.31 ± 7.16% and 4.48 ± 7.10%, respectively, as compared to other TTP and STP methods. The TTP methods based on later imaging time demonstrated fewer errors than the STP methods in kidney and tumor TIA. Imaging at 20 h-60 h and 40 h-200 h could simplify the dosimetry procedures with fewer TIA estimation errors.

Performance comparison of three passive samplers for monitoring of polar organic contaminants in treated municipal wastewater

Over the past decades, several types of passive samplers have been developed and used to monitor polar organic compounds in aquatic environments. These samplers use different sorbents and barriers to control the uptake into the sampler, but their performance comparison is usually not well investigated. This study aimed to directly compare the performance of three samplers, i.e., the Polar Organic Chemical Integrative Sampler (POCIS), the Hydrogel-based Passive Sampler (HPS, an upscaled version of o-DGT), and the Speedisk, on a diverse suite of pharmaceuticals, per- and polyfluoroalkylated substances (PFAS), and pesticides and their metabolites. The samplers were deployed side-by-side in the treated effluent of a municipal wastewater treatment plant for different exposure times. All samplers accumulated a comparable number of compounds, and integrative uptake was observed for most compounds detected up to 28days for POCIS, up to 14days for HPS, and up to 42days for Speedisk. In the integrative uptake phase, consistent surface-specific uptake was observed with a significant correlation between samplers (r≥0.76) despite differences in sampler construction, diffusion barrier, and sorbent material used. The low sampling rates compared to the literature and the low estimated overall mass transfer coefficient suggests that the water boundary layer was the main barrier controlling the uptake for all samplers. Although all devices provided comparable performance, Speedisk overcomes POCIS and HPS in several criteria, including time-integrative sampling over a long period and physical durability.

Statistical Post-Processing Method for Evaluating Bioaccumulation in Fish Due to Dietary Exposure in Japan.

In 2018, the dietary exposure bioaccumulation fish test of the Organization for Economic Co-operation and Development Test Guideline No. 305 was introduced into Japan's Chemical Substances Control Law. The Japanese government has adopted a single definitive testing criterion for the absence of high bioaccumulation: the growth-corrected kinetic dietary magnification factor (BMFKg) must be less than 0.007. The aim of this study was to decrease regulatory restrictions in order to increase newly developed chemical substances and their subsequent approval of their manufacture and import, i.e., the present study was motivated by concerns over the criterion being too restrictive, rather than scientific concerns, such as uncertainty in criterion. We used statistical post-processing to assess the possibility of expanding the criteria for not being highly bioaccumulative. Based on our results, we proposed the criterion that the test substance should be considered not highly bioaccumulative if the following two conditions are met: (1) The ratio of the maximum to the minimum measured 5% lipid-standardized biomagnification factor at the end of the uptake phase (BMF5%, n = 5) for the test substance and reference substance should be less than 3.0, and (2) For the measured BMF5% of the test substance (n = 5), the probability that the next (the sixth) BMF5% is below 0.0334 should exceed 95% based on statistical post-processing. It is worth noting that the BMF5% values should only be applied for non-ionizable lipid soluble compounds. Application of our suggested approach to Japan implies that the criterion for chemical substances that are not highly bioaccumulative in the dietary exposure bioaccumulation fish test would be increased from 0.007 to 0.0149.

Bioconcentration of carbamazepine, enalapril, and sildenafil in neotropical fish species.

Sewage effluents are the main source of entry of Human Pharmaceutical Active Ingredients (HPAIs) to surface water bodies. Carbamazepine (CBZ), psychiatric drug, enalapril (ENA) antihypertensive, and sildenafil (SIL), to treat erectile dysfunction, have been frequently detected in receiving wastewater and in wild fish species from Argentina. This study aimed to assess the bioconcentration of selected HPAIs in native fish species of the Del Plata Basin. In a first trial, the bioconcentration factors of CBZ, ENA, and SIL were obtained by exposing Cnesterodon decemmaculatus, respectively, to 135, 309, and 70μg/L during 96h. Then the bioconcentration kinetic of SIL was comparatively assessed in C. decemmaculatus and Piaractus mesopotamicus exposed, respectively, to 44.1 and 16.2μg/L during a one-week, followed by a four-day depuration phase. HPAIs concentrations in water and tissue were measured by HPLC-MS after 0.22μm filtration and direct injection or solid-liquid extraction, respectively. Bioconcentration factors obtained empirically (BCFe) for C. decemmaculatus were CBZ = 1.5, SIL = 1.4, and ENA = 0.007. Parameters estimated by the SIL bioconcentration kinetic model for C. decemmaculatus were: uptake rate constant (k1) = 5.5L/kg d, elimination rate constant during uptake phase (k2u) = 0.00175 d-1, maximum predicted tissue concentration (Ct(max)) = 138588μg/kg, estimated bioconcentration factor (BCFm) = 3143, lag time between the exposure and the first detection in tissue (tlag) = 0 d, elimination rate constant in the depuration phase (k2d) = 0.49 d-1 and half-life in the tissue (t1/2) = 1.4 d. The model parameters for P. mesopotamicus were k1: 7.3L/kg d, k2u: 0.0836 d-1, Ct(max): 1423μg/kg, BCFm: 88, tlag: 3.8 d in the uptake phase and k2d: 0.31 d-1 and t1/2: 2.3 d in the depuration phase. The reached conclusions were: 1) the bioconcentration capacity of CBZ and SIL are similar but around 200 times higher than ENA, 2) the time to reach the bioconcentration equilibrium for SIL is longer than 1week, then estimated BCFm are between 1 and 3 orders of magnitude higher than BCFe obtained after 96h exposure, but actual values need to be verified, 3) substantial differences (≈30 fold) were observed in the estimated BCF of SIL among species, indicating the need for further studies toward understanding such diversity to improve HPAIs ecological risk assessment worldwide.

Bioaccumulation, tissue distributions, and maternal transfer of perfluoroalkyl carboxylates (PFCAs) in laying hens

Laying hens were exposed to feeds spiked with a series of perfluoroalkyl carboxylates (PFCAs) ranging from perfluorobutanoic acid (C4) to perfluorooctadecanoic acid (C18) to investigate their bioaccumulation, tissue distribution, and maternal transfer. We found that PFCAs with longer carbon chains (>8) were more efficiently absorbed in the gastrointestinal tract than those with shorter chains (≤8), and that the rate of depuration varied inversely with the carbon chain length in a U-shaped pattern. Moreover, bioaccumulation potential increased with increasing carbon-chain length, except for C4. Distinct affinities were observed for specific carbon-chain PFCAs across various tissues, evident from their differential accumulation during both uptake and depuration phases. Specifically, C9 showed a higher affinity for serum and liver, C12 was more prevalent in yolk, C14 was notably abundant in the brain, and C18 was predominant in other tissues. Furthermore, the egg-maternal ratio (EMR) increased with increasing carbon-chain length from C7 to C11 and reached a plateau phase for C12 to C18. Our study also confirmed the key role of phospholipids in the tissue distribution and maternal transfer of long-chain PFCAs. This study sheds light on the interaction between PFCAs and biological tissues and reveals the toxicokinetic factors that influence the bioaccumulation of PFCAs. Further research is needed to identify the specific proteins or components that mediate the tissue-specific affinity for different carbon-chain lengths of PFCAs.

Cadmium-109 Internal Kinetics in Diamond Sturgeon Acipenser gueldenstaedtii are Strongly Influenced by Salinity, Exposure Pathway and History.

Cadmium-109 whole-body and internal biokinetics were experimentally investigated in critically endangered diamond sturgeon Acipenser gueldenstaedtii after uptake from water or food, in fresh (FW) and brackish (BW; 9‰) salinities typical of the Caspian Sea. Whole-body rates of uptake of 109Cd from water and subsequent depuration were quantified over 14 and 28days, respectively. Uptake was greater in FW than BW by a factor of 2.4, but depuration rates were similar in both salinities. In contrast, for the dietary (chironomid) exposure pathway 109Cd assimilation efficiencies (AEs) were higher in BW (13%) compared to FW (9.5%). Head (including gills) or digestive tract were major repositories of 109Cd following aqueous and dietary exposures, respectively, including both uptake and depuration phases. The point-of-entry of 109Cd into the body was also a major and persistent determiner of its subsequent internal distribution. For aqueous exposures, the internal distributions of 109Cd changed appreciably during depuration with increased activity concentrations in some body components, which again varied with salinity. Increased salinity appreciably enhanced the percentage distributions and activity concentrations of 109Cd in the liver, kidney and digestive tract, which are typically most pathologically altered by elevated Cd exposure. For dietary exposure, increased salinity also enhanced 109Cd activity concentrations in most body components. The results repeatedly indicate the important role of salinity in the whole-body and internal biokinetics of 109Cd in A. gueldenstaedtii, a representative of both a phylogenetically distinct and most endangered family of fishes.

Targeting Efferocytosis in Inflammaging.

Rapid removal of apoptotic cells by phagocytes, a process known as efferocytosis, is key for the maintenance of tissue homeostasis, the resolution of inflammation, and tissue repair. However, impaired efferocytosis can result in the accumulation of apoptotic cells, subsequently triggering sterile inflammation through the release of endogenous factors such as DNA and nuclear proteins from membrane permeabilized dying cells. Here, we review the molecular basis of the three key phases of efferocytosis, that is, the detection, uptake, and degradation of apoptotic materials by phagocytes. We also discuss how defects in efferocytosis due to the alteration of phagocytes and dying cells can contribute to the low-grade chronic inflammation that occurs during aging, described as inflammaging. Lastly, we explore opportunities in targeting and harnessing the efferocytic machinery to limit aging-associated inflammatory diseases.

Improving Product Safety for Edible Insects: Toxicokinetics of Hg in Tenebrio molitor and Hermetia illucens

Sustainability, circularity, and Zero Waste policies are timely concepts for policy development and strategies in the European Union (EU) and other global regions. Insects can likely become key players in the bioconversion of waste to valuable material and promise one solution to achieve diverse societal goals. Insects further present strategic opportunities as food products; however, it is necessary to understand how insects accumulate and eliminate priority contaminants from different substrates where they can be reared. In the present study, we expanded beyond previous work with mercury (Hg) to examine bioaccumulation kinetics in Tenebrio molitor (YMW) and Hermetia illucens (BSF). Two-phase bioaccumulation assays, with an uptake (contaminated Hg substrate) and elimination phase (clean substrate), followed by toxicokinetic modeling, showed that both insects have a high capacity to regulate Hg, often reaching an internal steady-state concentration at level responding on the substrate concentration of Hg. Of importance for product safety, both insects quickly eliminated Hg after being transferred to clean substrate. Specifically, BSF eliminated half of the accumulated Hg in approximately 1 day (after 5 days of Hg exposure) and YMW in 4–5 days (after 21 days of Hg exposure). These results provide crucial product safety information for insect producers using possibly contaminated substrates, specifically informing the amount of time for Hg depuration prior to processing and commercialization for food and feed.

Modeling identifies variability in SARS-CoV-2 uptake and eclipse phase by infected cells as principal drivers of extreme variability in nasal viral load in the 48 h post infection

The SARS-CoV-2 coronavirus continues to evolve with scores of mutations of the spike, membrane, envelope, and nucleocapsid structural proteins that impact pathogenesis. Infection data from nasal swabs, nasal PCR assays, upper respiratory samples, ex vivo cell cultures and nasal epithelial organoids reveal extreme variabilities in SARS-CoV-2 RNA titers within and between the variants. Some variabilities are naturally prone to clinical testing protocols and experimental controls. Here we focus on nasal viral load sensitivity arising from the timing of sample collection relative to onset of infection and from heterogeneity in the kinetics of cellular infection, uptake, replication, and shedding of viral RNA copies. The sources of between-variant variability are likely due to SARS-CoV-2 structural protein mutations, whereas within-variant population variability is likely due to heterogeneity in cellular response to that particular variant. With the physiologically faithful, agent-based mechanistic model of inhaled exposure and infection from (Chen et al., 2022), we perform statistical sensitivity analyses of the progression of nasal viral titers in the first 0-48h post infection, focusing on three kinetic mechanisms. Model simulations reveal shorter latency times of infected cells (including cellular uptake, viral RNA replication, until the onset of viral RNA shedding) exponentially accelerate nasal viral load. Further, the rate of infectious RNA copies shed per day has a proportional influence on nasal viral load. Finally, there is a very weak, negative correlation of viral load with the probability of infection per virus-cell encounter, the model proxy for spike-receptor binding affinity.

Cell-based BSEP trans-inhibition: A novel, non-invasive test for diagnosis of antibody-induced BSEP deficiency

<h2>Abstract</h2><h3>Background&Aims</h3> Antibody-induced bile salt export pump deficiency (AIBD) is an acquired form of intrahepatic cholestasis, which may develop following orthotopic liver transplantation (OLT) for progressive familial intrahepatic cholestasis type 2 (PFIC-2). Approximately 8-33% of transplanted PFIC-2 patients develop bile salt export pump (BSEP) antibodies which trans-inhibit this bile salt transporter from the extracellular, biliary side. AIBD is diagnosed by demonstration of BSEP-reactive and -inhibitory antibodies in patient serum. We developed a cell-based test directly measuring BSEP trans-inhibition by antibodies in serum samples to confirm AIBD diagnosis. <h3>Methods</h3> Sera from healthy controls and cholestatic Non-AIBD or AIBD cases were tested I) for anti-canalicular reactivity by immunofluorescence (IF) staining of human liver cryosections, II) for anti-BSEP reactivity by IF staining of HEK293 cells expressing BSEP-EYFP and immunodetection of BSEP-EYFP on Western blot, and III) for BSEP trans-inhibition using HEK293 cells stably expressing NTCP-mCherry and BSEP-EYFP. The trans-inhibition test uses [³H]-taurocholate as substrate and is divided into an uptake phase dominated by NTCP followed by BSEP-mediated export. For functional analysis, sera were bile salt depleted. <h3>Results</h3> We found BSEP trans-inhibition by seven sera containing anti-BSEP antibodies, but not by five cholestatic or nine control sera, all lacking BSEP reactivity. Prospective screening of a post-OLT PFIC-2 patient showed seroconversion to AIBD and the novel test method allowed monitoring of treatment response. Notably, we identified a post-OLT PFIC-2 patient with anti-BSEP antibodies yet without BSEP trans-inhibition activity, in line with asymptomatic presentation at serum sampling. <h3>Conclusions</h3> Our cell-based assay is the first direct functional test for AIBD and allows confirmation of diagnosis as well as monitoring under therapy. We propose an updated workflow for AIBD diagnosis including this functional assay. <h3>Lay Summary</h3> Antibody-induced BSEP deficiency (AIBD) is a potentially serious complication that may affect PFIC-2 patients after liver transplantation. To improve its early diagnosis and thus immediate treatment, we developed a novel functional assay to confirm AIBD diagnosis using patient's serum and propose an updated diagnostic algorithm for AIBD.

The Role of Calcium in Augmenting the Efficacy of Primary Homeostasis after Hemorrhagic Shock

Background: Traumatic Hemorrhagic Shock (THS) causes hypocalcemia (Ca). This study assesses how low Ca affects platelet (Plt) function. Study Design: A research registry was queried for 555 studies on 341 patients (pts) with THS requiring 14.2 units RBC, 8.4 units FFP, 3.6 units Plt, and 18.9 mEq Ca by end of O.R. (7.3 hours). Factors analyzed included Plt count, Ca, Bleeding Time (BT), Plt aggregation with ADP (PAadp) and collagen (PAcol), including Max PA, T1/2 PA, and rate PA, plus platelet release factors, beta thromboglobulin (BTG) and Plt PF-4. Studies were made in O.R. (26 pts) during fluid uptake phase II at 29 hours (216 pts), during fluid mobilization phase III at 56 hours (283 pts), and as outpatients (30 pts). Results: THS caused low Plts (1x10 SD) in O.R (107 ± 43), in phase II (96 ± 35), and in phase III (105 ± 59) and low Ca in O.R. (1.6 ± 0.3), in phase II (1.8 ± 0.2), and in phase III (2.1 ± 0.2). Low Plt and Ca correlated (p=&lt;0.05) with PAadp and PAcol in O.R., phase II, and phase III. Ca correlated directly with Plt count, aggregation, BTG and PF-4, and inversely with BT. All outpatient studies were normal. Conclusion: THS causes Ca which leads to low Plt and impaired function. Routine Ca supplementation is recommended for THS.

The deep radiomic analytics pipeline.

Radiomics refers to the process of extracting useful imaging features from radiological data. Conventional radiomics like standard uptake value, intensity histograms, or phase images involve hand-crafted (manual) or automated regions of interest (computer generated), however, artificial intelligence (AI) segmentation (AI-augmented radiomics) has recently emerged. Radiomic feature extraction extends image insights beyond simply data quantitation and provides additional insights to aid semantic reporting. Deeper layers of a convolutional neural network produce more abstract radiomic features that are referred to as deep radiomics. The application of radiomics in veterinary radiology is already firmly entrenched using hand-crafted and automated computer-generated radiomic features in X-ray, nuclear medicine, CT, ultrasound, and MRI. There is an opportunity for veterinary radiology to capitalize on advances in AI, machine learning, and deep learning to enrich imaging interpretation using deep radiomic feature extraction. This manuscript aims to provide a general understanding of radiomics and deep radiomics, and to arm readers with the vernacular to progress discussion and development of deep radiomics in veterinary imaging.