Species Differences Research Articles

Spatially-resolved cluster dynamics modeling of irradiation growth

We develop here a spatially resolved, three-dimensional continuum model coupling cluster dynamics (SR-CD) and crystal plasticity to investigate irradiation growth in zirconium. The model uses scale separation to divide the population of the irradiation cluster into mobile and immobile families. Small interstitial and vacancy clusters are modeled using anisotropic reaction–diffusion equations. Among the immobile clusters, an atomistically-informed vacancy cluster to vacancy loop transition is taken into account. The coupling between the evolution equation of CD and the plastic deformation of the material is two-fold, with stress-informed bias factors and local inelastic strains computed from the evolution of the evolving cluster population. The numerical implementation of the model utilizes the finite element method to analyze both single-crystal and polycrystalline samples. The growth strains that are computed align well with the experimental data provided by Carpenter for single-crystal Zr. Furthermore, the transformation of a vacancy cluster into a complete vacancy loop, occurring at a size of 14 nm, is in agreement with experimental observations and atomistic simulations. The density, size, and growth rate of the dislocation loops, denoted as 〈c〉 and 〈a〉, also exhibit good agreement with transmission electron microscopy (TEM) analysis of irradiated Zr and its alloys. Our findings demonstrate that there is a spatial correlation between the growth of these dislocation loops and growth strains, significantly influenced by the crystal size. To explain the expansion of the 〈a〉 axis and the contraction of the 〈c〉 axis in irradiated Zr, it is necessary to consider the diffusion anisotropy difference (DAD) of mobile interstitial species. We show that the PWR Kearns parameters, specifically fr = 0.63, ft = 0.32, fa = 0.05, confer enhanced irradiation resistance to Zr along the principal directions when compared to single crystals. Additionally, reducing the grain size to nanograins further enhances the resistance to irradiation-induced growth, particularly along the direction with the highest volume fraction of basal poles [0001].

Single-cell and spatial transcriptomics analysis of human adrenal aging

ObjectiveThe human adrenal cortex comprises three functionally and structurally distinct layers that produce layer-specific steroid hormones. With aging, the human adrenal cortex undergoes functional and structural alteration or “adrenal aging”, leading to the unbalanced production of steroid hormones. Given the marked species differences in adrenal biology, the underlying mechanisms of human adrenal aging have not been sufficiently studied. This study was designed to elucidate the mechanisms linking the functional and structural alterations of the human adrenal cortex. MethodsWe conducted single-cell RNA sequencing and spatial transcriptomics analysis of the aged human adrenal cortex. ResultsThe data of this study suggest that the layer-specific alterations of multiple signaling pathways underlie the abnormal layered structure and layer-specific changes in steroidogenic cells. We also highlighted that macrophages mediate age-related adrenocortical cell inflammation and senescence. ConclusionsThis study is the first detailed analysis of the aged human adrenal cortex at single-cell resolution and helps to elucidate the mechanism of human adrenal aging, thereby leading to a better understanding of the pathophysiology of age-related disorders associated with adrenal aging.

160 Awardee Talk: Impact of indigestible protein on coccidiosis and meat and bone meal autoclaving time-related reduction in amino acid digestibility for broiler chickens and growing pigs

Abstract Meat and bone meal (MBM) is a byproduct of the processing of livestock in the rendering industry. Because of its highly digestible protein and balanced amino acid profile, MBM is a valuable protein source in swine and poultry diets. Rendering may reduce amino acid digestibility of MBM because of heat damage to MBM, and the effect of heat damage on amino acid digestibility in MBM may be different between broiler chickens and growing pigs. For this reason, the objective of this study was to determine the effect of autoclaving on standardized ileal digestibility (SID) of amino acid in MBM fed to broiler chickens and growing pigs. Meat and bone meal from the same batch was autoclaved at 134°C for 0, 30, 60, 90, 120, 150, or 180 min, generating 7 samples. Eight experimental diets consisted of 7 diets containing each MBM sample as a sole source of nitrogen and a nitrogen-free diet. In Exp. 1, male broiler chickens [n = 656; initial body weight (BW) = 777 ± 69 g] at d 18 post hatching were assigned to the 8 diets in a randomized complete block design with BW as a blocking factor. On d 23, birds were euthanized by CO2 asphyxiation and dissected for the collection of ileal digesta. In Exp. 2, barrows (n = 16; initial BW = 25.7 ± 1.0 kg) surgically fitted with T-cannulas at the distal ileum were allotted to duplicate 8 × 4 incomplete Latin square design with the 8 diets and 4 periods. Each experimental period consisted of 5-d adaptation and 2-d ileal digesta collection periods. Data for Exp. 1 and 2 were pooled as a 2 × 7 factorial treatment arrangement with the effects of species (i.e., broiler chickens or pigs) and autoclaving time (i.e., 0 to 180 min). The basal endogenous losses of most AA in growing pigs were greater (P < 0.05) compared with broiler chickens. The SID of nitrogen and tryptophan in MBM linearly decreased (P < 0.05) in both species with increasing autoclaving time. An interaction between species and quadratic effect of autoclaving time was observed (P < 0.05) in SID of arginine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, and valine in MBM. Specifically, the SID of lysine both linearly and quadratically decreased (P < 0.05) in broiler chickens (85.0 to 56.0%), whereas the SID of lysine only linearly decreased (P < 0.05) in growing pigs (79.5 to 46.7%) as autoclaving time increased. In conclusion, given the species differences in amino acid digestibility response to the severity of heat damage, target species should be considered when using amino acid digestibility value of MBM in diet formulation.

Identification of UDP-glucuronosyltransferase (UGT) isoforms involved in the metabolism of Chlorophenols (CPs)

Chlorophenols (CPs) are a group of pollutants that pose a great threat to the environment, they are widely used in industrial and agricultural wastes, pesticides, herbicides, textiles, pharmaceuticals and plastics. Among CPs, pentachlorophenol was listed as one of the persistent organic pollutants (POPs) by the Stockholm convention. This study aims to identify the UDP-glucosyltransferase (UGT) isoforms involved in the metabolic elimination of CPs. CPs’ mono-glucuronide was detected in the human liver microsomes (HLMs) incubation mixture with co-factor uridine-diphosphate glucuronic acid (UDPGA). HLMs-catalyzed glucuronidation metabolism reaction equations followed Michaelis-Menten or substrate inhibition type. Recombinant enzymes and chemical reagents inhibition experiments were utilized to phenotype the main UGT isoforms involved in the glucuronidation of CPs. UGT1A6 might be the major enzyme in the glucuronidation of mono-chlorophenol isomer. UGT1A1, UGT1A6, UGT1A9, UGT2B4 and UGT2B7 were the most important five UGT isoforms for metabolizing the di-chlorophenol and tri-chlorophenol isomers. UGT1A1 and UGT1A3 were the most important UGT isoforms in the catalysis of tetra-chlorophenol and pentachlorophenol isomers. Species differences were investigated using rat liver microsomes (RLMs), pig liver microsomes (PLMs), dog liver microsomes (DLMs), and monkey liver microsomes (MyLMs). All these results were helpful for elucidating the metabolic elimination and toxicity of CPs.

The vedotin antibody-drug conjugate payload drives platform-based nonclinical safety and pharmacokinetic profiles.

Nonclinical safety and pharmacokinetic data for MMAE and 14 vedotin ADCs were evaluated to determine patterns of toxicity, consistency of pharmacokinetic results, and species differences between rats and monkeys. Most nonclinical toxicities were antigen-independent, common across ADCs, and included hematologic, lymphoid, and reproductive toxicity related to MMAE pharmacology. Hematologic toxicity was the dose-limiting or predominant toxicity for the majority of vedotin ADCs in both species. Tissue expression of the targeted antigen of an ADC rarely correlated with dose-limiting toxicity (DLT); only two ADCs had antigen-dependent skin DLTs. For two additional ADCs, antigen-dependent delivery of MMAE in the bone marrow may have exacerbated the antigen-independent hematologic DLT. The highest tolerated doses and pharmacokinetics were similar within a given species, with rats tolerating higher doses than monkeys. Studies longer than one month in duration detected the same or fewer toxicities than one-month studies and had no additional findings that affected the human risk assessment. These data support opportunities to streamline ADC toxicity assessments without compromising human starting dose selection or target organ identification.

Comparison of bacterial species and clinical outcomes in patients with diabetic hand infection in tropical and nontropical regions.

Hand infection is a rare complication in patients with diabetes. Its clinical outcomes depend on the severity of hand infection caused by bacteria, but the difference in bacterial species in the regional disparity is unknown. The purpose of this study was to explore the influence of tropical and nontropical regions on bacterial species and clinical outcomes for diabetic hand. A systematic literature review was conducted using PubMed, EMBASE, Web of Science, and Google Scholar. Moreover, the bacterial species and clinical outcomes were analyzed with respect to multicenter wound care in China (nontropical regions). Both mixed bacteria (31.2% vs. 16.6%, p = 0.014) and fungi (7.5% vs. 0.8%, p = 0.017) in the nontropical region were significantly more prevalent than those in the tropical region. Staphylococcus and Streptococcus spp. were dominant in gram-positive bacteria, and Klebsiella, Escherichia coli, Proteus and Pseudomonas in gram-negative bacteria occupied the next majority in the two regions. The rate of surgical treatment in the patients was 31.2% in the nontropical region, which was significantly higher than the 11.4% in the tropical region (p = 0.001). Although the overall mortality was not significantly different, there was atendencytobeincreased in tropical regions (6.3%) compared with nontropical regions (0.9%). However, amputation (32.9% vs. 31.3%, p = 0.762) and disability (6.3% vs. 12.2%, p = 0.138) were not significantly different between the two regions. Similar numbers of cases were reported, and the most common bacteria were similar in tropical and nontropical regions in patients with diabetic hand. There were more species of bacteria in the nontropical region, and their distribution was basically similar, except for fungi, which had differences between the two regions. The present study also showed that surgical treatment and mortality were inversely correlated because delays in debridement and surgery can deteriorate deep infections, eventually leading to amputation and even death.

Parametrization of biological assumptions to simulate growth of tree branching architectures.

Modeling and simulating the growth of the branching architecture of tree species remains a challenge. With existing approaches, we can reconstruct or rebuild the branching architectures of real tree species, but the simulation of the growth process remains unresolved. First, we present a tree growth model to generate branching architectures that resemble real tree species. Second, we use a quantitative morphometric approach to infer the shape similarity of the generated simulations and real tree species. Within a functional-structural plant model (FSPM), we implement a set of biological parameters that affect the branching architecture of trees. By modifying the parameter values, we aim to generate basic shapes of spruce, pine, oak, and poplar. Tree shapes are compared using geometric morphometrics of landmarks that capture crown and stem outline shapes. Five biological parameters, i.e. xylem flow, shedding rate, proprioception, gravitysense, and lightsense, most influenced tree branching and their adjustments led to the generation of different spruce, pine, oak, and poplar shapes. The largest effect was attributed to gravity, as phenotypic responses to this effect resulted in different growth directions of gymnosperm and angiosperm branching architectures. Since we were able to obtain branching architectures that resemble real tree species by adjusting only a few biological parameters, our model is extendable to other tree species. Furthermore, the model will also allow the simulation of structural tree-environment interactions. Our simplifying approach to shape comparison between tree species, landmark geometric morphometrics, showed that even the crown-trunk outlines capture species differences based on their contrasting branching architectures.

Development of a UAV-borne sorbent tube sampler and its application on the vertical profile measurement of volatile organic compounds

Vertical detection of volatile organic compounds (VOCs) is essential to expend our understanding of the distribution characteristics of VOCs and improve the predictive ability of existing air quality models. In this work, we report the development of a sorbent tube sampler based on an unmanned aerial vehicle (UAV) platform. Vertical profile measurement of VOCs with a vertical resolution of 25 m was achieved. The sampler consists of five lightweight VOC sorbent tubes and a 5-way solenoid valve, making it available for collecting five atmospheric VOC samples in a single flight with a time response of less than 30 min. The sampler weighed ∼ 1.45 kg and had dimensions of 240 mm × 220 mm × 100 mm with small penetration loss (< 10%) under 4-liter sampling conditions (flow rate of 200 mL/min). Commercialized SUMMA canisters were used as experimental controls to investigate the possible loss of self-made sampler for target compounds in the same sampling process. Comparison experiment on the ground showed that the concentration differences for all VOC species were lower than 0.14 μg/m3, proving the good reliability for VOCs measurements using sorbent tube sampler. The UAV platform also incorporated online instruments for meteorological parameters and O3 measurement. The sampler was successfully applied to characterize the vertical profiles of VOCs up to 100 m in October 2023 in the Huaihe River Basin of China. The UAV platform and the sorbent tube sampler demonstrate good performance and will be a valuable and reliable tool for vertical VOCs measurement.

CULTURABLE ENDOSYMBIOTIC BACTERIA FROM THE INDIAN LAC INSECT, KERRIA LACCA (KERR)

The Indian lac insect, Kerria lacca (Kerr) (Coccoidea: Tachardiidae) is a commercially important phytosuccivorous and sessile scale insect. Lac insects are cultured on suitable host plants in India and some Southeast Asian countries to produce lac. The lac insect harbours a number of endosymbionts. Isolation of culturable microbial endosymbionts and their identification through 16S rRNA has revealed sex and host-related differences of microbial species. Bacillus boroniphilus, Enterobacter cloacae and Staphylococcus sp. were found only in the lac insects reared on the plant host Cajanus cajan, whereas Bacillus firmus, Lysinibacillus xylanilyticus, Bacillus horneckiae and Bacillus velezensis were recorded only from Flemingia macrophylla. B. firmus and L. xylanilyticus were female-specific and B. horneckiae and B. velezensis were male-specific with Flemingia macrophylla as host; E. cloacae was female-specific and Bacillus boroniphilus and Staphylococcus sp. were male specific with C. cajan. Biochemical characteristics of the isolates, their genetic relationship with their taxonomic kin and their probable role, based on the information available about these endosymbionts in other hosts, have been studied.

Analgesic and Anti-Inflammatory Activity: A Comprehensive Review on Invitro and In Vivo Screening Methods

Modern pharmacotherapy includes analgesic and anti-inflammatory medicines as essential components to relieve pain and inflammation brought on by a variety of medical diseases. Robust screening techniques are essential for the identification of possible candidates with appropriate safety and effectiveness profiles in the search and development of new analgesic and anti-inflammatory medications. This study looks at the many screening methods used in preclinical studies to assess new drugs' analgesic and anti-inflammatory quality. Conventional techniques like the tail flick, hot plate, and writhing’s tests measure analgesic activity by having animals respond to unpleasant stimuli. Comparably, anti-inflammatory activity is frequently assessed using assays like the cotton pellet granuloma test, which gauges tissue granuloma formation, and the carrageenan-induced paw edema model, which measures inflammation. These traditional techniques offer insightful information about the pharmacological effects of test substances. Despite the wide range of screening techniques available, each strategy has advantages and disadvantages. Preclinical studies are more reliable and have higher predictive value when various assays and techniques are integrated into a tiered screening strategy. Furthermore, the successful translation of preclinical findings to human applications depends on taking into account translational variables including species differences and clinical relevancies. As a result, choosing the right screening techniques is critical to the effective identification and characterization of new analgesic and anti-inflammatory drugs.

Anthraquinone biocolourant dermocybin is metabolized whereas dermorubin is not in in vitro liver fractions and recombinant metabolic enzymes.

Fungal anthraquinones dermocybin and dermorubin are attractive alternatives for synthetic dyes but their metabolism is largely unknown. We conducted a qualitative in vitro study to identify their metabolism using human liver microsomes and cytosol, as well as recombinant human cytochrome P450 (CYP), UDP-glucuronosyltransferase (UGT) and sulfotransferase (SULT) enzymes. Additionally, liver microsomal and cytosolic fractions from rat, mouse and pig were used. Following incubations of the biocolourants with the enzymes in the presence of nicotinamide adenine dinucleotide phosphate, UDP-glucuronic acid, 3'-phosphoadenosine-5'-phosphosulfate (PAPS) or S-adenosyl methionine (SAM) to enable CYP oxidation, glucuronidation, sulfonation or methylation, we observed several oxidation and conjugation metabolites for dermocybin but none for dermorubin. Human CYP1A1, 1A2, 1B1, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4 and 3A7 catalysed dermocybin oxidation. The formation of dermocybin glucuronides was catalysed by human UGT1A1, 1A3, 1A7, 1A8, 1A9, 1A10 and 2B15. Human SULT1B1, 1C2 and 2A1 sulfonated dermocybin. Dermocybin oxidation was faster than conjugation in human liver microsomes. Species differences were seen in dermocybin glucuronidation between human, rat, mouse and pig. In conclusion, many CYP and conjugation enzymes metabolized dermocybin, whereas dermorubin was not metabolized in human liver fractions in vitro. The results indicate that dermocybin would be metabolized in humans in vivo.

Antibiotic susceptibility of Streptococcus species that cause pharyngitis in children

Background: Group A Streptococcus (GAS) significantly impacts global health, especially among children aged 5–14 years, causing approximately 288 million episodes of sore throat annually and resulting in around 100,000 disability-adjusted life years (DALYs) each year. GAS infections are particularly prevalent in developing countries. Objective: To assess the patterns of antibiotic resistance of Streptococcus spp. that cause pharyngitis in children. Patients and Methods: It was collected 100 pediatric patients. It was taken throat swabs for Streptococcus species culture and antibiotic sensitivity test. In addition, it was recorded the data of patient’s age and gender. Results: It was shown in this study that an average age of 4.4 ± 3.7 years for pharyngitis in children. In this study was diagnosed that Streptococcus agalactiae (24%) and Streptococcus pyogenes (18%) as the common causes Streptococcus species causes pharyngitis in children. It was noted that the differences in bacterial species according to the patient’s ages. Antibiotic sensitivity testing determined that there are high sensitivity of S. pneumoniae and S. parasanguinis to multiple antibiotics. In addition, it was showed resistance of S. pyogenes to Metronidazole and Azithromycin. Conclusion: In this study, it was found that the risk age for pharyngitis in children is four years old. Streptococcus agalactiae and Streptococcus pyogenes are the common bacterial causes for this infection. In addition, it was shown that S. pneumoniae appeared a high sensitivity to Ceftriaxone, Ciprofloxacin, and Meropenem, and S. parasanguinis was sensitive to Azithromycin, Cefixime, and Cefotaxime, and S. pyogenes showed a high sensitivity to Meropenem and Imipenem.

Scouts vs. usurpers: alternative foraging strategies facilitate coexistence between neotropical Cathartid vultures

Understanding how diverse assemblages of scavengers can coexist on shared ecological resources is a fundamental challenge in community ecology. However, current approaches typically focus on behaviour at carcass provisioning sites, missing how important differences in movement behaviour and foraging strategies can facilitate sympatric species coexistence. Such information is particularly important for vultures – obligate scavengers representing the most endangered avian foraging guild. Their loss from ecosystems can trigger trophic cascades, mesopredator release and disease outbreaks. We provide the first‐ever analyses of GPS location data from wild King Vultures Sarcoramphus papa and Greater Yellow‐headed Vultures Cathartes melambrotus, coupled with trait data (from both wild‐living and museum specimens) and visitation data from camera traps deployed at provisioned carcasses, to characterize vulture flight behaviour and strategies in the Peruvian Amazon. We found marked species differences in several key movement characteristics, including: King Vultures having home‐ranges five times larger, average flight heights four times greater and ground speeds 40% faster than those of Greater Yellow‐headed Vultures. Despite these differences, both species flew similar distances each day (on average), probably due to King Vultures taking 50% fewer flights and spending 40% less time in the air per day. Consistent with these patterns, King Vulture body mass was more than double that of the Greater Yellow‐headed Vulture, with a substantially larger hang wing index (a measure of long‐distance flight efficiency). At carcasses, Greater Yellow‐headed Vultures typically arrived first but were rapidly outnumbered by both King and Black Vultures Coragyps atratus. We find that the movement behaviour of obligate apex scavengers in the western Amazon is linked to their ability to coexist – Greater Yellow‐headed Vultures, a smaller stature ‘scouting’ species adapted to fly low, forage early and arrive first at carcasses, are ultimately displaced by larger‐bodied, wider ranging King Vultures at large ephemeral carrion resources. Expansion of future GPS tracking initiatives could facilitate the exploration of direct facultative interactions from animal movement data and give further insight into how diverse communities assemble and interact.

Exploring precision-cut liver slices for comparative xenobiotic metabolism profiling in swine and cattle

In vitro systems are useful tools for unravelling species differences in xenobiotic metabolism. The current work aimed to validate the technique of precision-cut liver slices (PCLS) for comparative studies on xenobiotic metabolism in swine and cattle. PCLS from swine (n = 3) and cattle (n = 3) were produced using a Brendel-VitronTM Tissue Slicer and cultured for 6 h. Tissue viability was preserved throughout the whole culture period. Metabolic viability was evaluated using the anthelmintics albendazole (ABZ) and fenbendazole (FBZ) as model drugs, as well as other substrates of hepatic monooxygenases: benzydamine (BZ) N-oxygenase (FMO-dependent), and the O-dealkylations of 7-ethoxyresorufin (EROD, CYP1A1-dependent) and 7-methoxyresorufin (MROD, CYP1A2-dependent). ABZ S-oxygenation resulted 6-fold (cattle) and 13.6-fold (swine) higher (p = 0.001) compared to FBZ S-oxygenation. Similar BZ N-oxygenation and EROD activities were observed in PCLS cultures from both species. MROD was 2.5-fold higher (p = 0.033) in swine than in cattle. Similarly, ABZ S-oxygenation was 1.7-fold higher (p = 0.0002) in swine than in cattle. Conversely, a 82% higher (p = 0.0003) rate of FBZ S-oxygenation was evidenced in PCLS cultures from cattle compared to those from swine. Overall, this work shows that PCLS cultures are useful to obtain relevant information on species differences in xenobiotic metabolism.

Impact of four different extraction methods and three different reconstitution solvents on the untargeted metabolomics analysis of human and rat urine samples

Unsuitable sample preparation may result in loss of important analytes and consequently affect the outcome of untargeted metabolomics. Due to species differences, different sample preparations may be required within the same biological matrix.The study aimed to compare the in-house sample preparation method for urine with methods from literature and to investigate the transferability of sample preparation from human urine to rat urine. A total of 12 different conditions for protein precipitation were tested, combining four different extraction solvents and three different reconstitution solvents using an untargeted liquid-chromatography high resolution mass spectrometry (LC-HRMS) metabolomics analysis. Evaluation was done based on the impact on feature count, their detectability, as well as the reproducibility of selected compounds.Results showed that a combination of methanol as extraction and acetonitrile/water (75/25) as reconstitution solvent provided improved results at least regarding the total feature count. Additionally, it was found that a higher amount of methanol was most suitable for extraction of rat urine among the tested conditions. In comparison, human urine requires significantly less volume of extraction solvent.Overall, it is recommended to systematically optimize both, the extraction method, and the reconstitution solvent for the used biofluid and the individual analytical settings.

Social buffering of behavioural stress response in two fish species, Nile tilapia (Oreochromis niloticus) and koi carp (Cyprinus carpio)

AbstractThe presence of conspecifics plays a crucial role in mitigating stress responses in social teleost species and holds potential for enhancing welfare in captive fish. While studies on social buffering effects have typically focussed on single species, marked interspecific differences can exist. Here, we conducted an analysis of social buffering of stress response in two of the most extensively farmed fish, the Nile tilapia and the koi carp. Subjects were exposed to a behavioural stress response assay (open‐field test) in three conditions simulating increasing levels of social enrichment: isolation, pairs, or shoals of five fish. We obtained five stress indicators from the assay: thigmotaxis, freezing, activity, erratic movements and interindividual distance in conditions with more than one fish. In both species, erratic movements significantly decreased with increasing levels of social enrichment, suggesting a similar social buffering effect. However, other indicators revealed species differences. Koi carp, but not Nile tilapia, showed a socially‐mediated reduction in thigmotaxis, whereas Nile tilapia, but not Koi carp, showed a socially‐mediated reduction in freezing behaviour. Furthermore, social enrichment determined opposite effects on the activity of the two species: Nile tilapia were more active as group size increased, whereas the opposite trend was found in koi carp. Finally, Nile tilapia showed increased interindividual distance with increasing social group size, whereas no changes were observed for koi carp. Our study indicates that the buffering effects of social enrichment on the behavioural stress response do not completely overlap between different fish species, highlighting the importance of developing finely‐tuned species‐specific enrichments and welfare indicators.

A roadmap towards a human-centric safety assessment of advanced therapy medicinal products

Advanced therapy medicinal products (ATMPs) are among the most complex pharmaceuticals with high human specificity. Species differences severely limit the clinical relevance of in vivo data. We conducted interviews with stakeholders involved in ATMP development about their perspective on the use of in vivo studies, the perceived hurdles and associated potential solutions regarding non-clinical development of ATMPs. In total, 17 stakeholders from 9 different countries were interviewed. A workshop was held with key stakeholders to further discuss major topics identified from the interviews. Conducting in vivo studies remains the status quo for ATMPs development. The hurdles identified included determining the amount of information required before clinical entry and effective use of limited human samples to understand a treatment or for clinical monitoring. A number of key points defined the need for future in vivo studies as well as improved application and implementation of New Approach Methodology (NAM)-based approach for products within a well-known modality or technology platform. These included data transparency, understanding of the added value of in vivo studies, and continuous advancement, evaluation, and qualification of NAMs. Based on the outcome of the discussions, a roadmap with practical steps towards a human-centric safety assessment of ATMPs was established.

Differences in diacylglycerol acyltransferases expression patterns and regulation cause distinct hepatic triglyceride deposition in fish

Triglyceride (TAG) deposition in the liver is associated with metabolic disorders. In lower vertebrate, the propensity to accumulate hepatic TAG varies widely among fish species. Diacylglycerol acyltransferases (DGAT1 and DGAT2) are major enzymes for TAG synthesis. Here we show that large yellow croaker (Larimichthys crocea) has significantly higher hepatic TAG level than that in rainbow trout (Oncorhynchus mykiss) fed with same diet. Hepatic expression of DGATs genes in croaker is markedly higher compared with trout under physiological condition. Meanwhile, DGAT1 and DGAT2 in both croaker and trout are required for TAG synthesis and lipid droplet formation in vitro. Furthermore, oleic acid treatment increases DGAT1 expression in croaker hepatocytes rather than in trout and has no significant difference in DGAT2 expression in two fish species. Finally, effects of various transcription factors on croaker and trout DGAT1 promoter are studied. We find that DGAT1 is a target gene of the transcription factor CREBH in croaker rather than in trout. Overall, hepatic expression and transcriptional regulation of DGATs display significant species differences between croaker and trout with distinct hepatic triglyceride deposition, which bring new perspectives on the use of fish models for studying hepatic TAG deposition.

Sex, Age, and Species Differences of Perfluorooctanoic Acid Modeled by Flow- versus Permeability-limited Physiologically-Based Pharmacokinetic Models.

This study aimed to investigate sex, age, and species differences of perfluorooctanoic acid (PFOA) using physiologically-based pharmacokinetic (PBPK) models in rats and humans. PBPK models were generally developed as either flow- or permeability-limited models. The flow-limited model is cost-effective and allows for human PK prediction through simple allometric scaling, while the permeability-limited model can incorporate detailed information on the disposition process through in vitro-in vivo extrapolation (IVIVE). PFOA was administered via oral or intravenous administration with 5mg/kg in male and female rats of different ages and the data was used to develop the PBPK models. Our results showed that both models successfully captured sex differences in rats, while only the flow-limited model with male rats and the permeability-limited model with both male and female rats provided comparable predictions in the human clinical study. More than the flow-limited model, the permeability-limited model effectively explained sex differences in rats and species differences through IVIVE. Additionally, the ontogeny-based mechanistic description of PFOA disposition enabled the interpretation of age- and sex-dependent pharmacokinetics. Although the flow-limited PBPK model lacked mechanistic interpretability compared to the permeability-limited model, it demonstrated reliable human prediction through simple allometric scaling. In conclusion, the permeability PBPK model could interpret age, sex, and species differences and it could improve the accuracy of human prediction.

Inhibition kinetics of acetylcholinesterase and butyrylcholinesterase from various species by 2-(2-cresyl)-4H-1,3,2-benzodioxaphosphorin-2-oxide (CBDP)

The aerotoxic syndrome has been associated with exposure to tricresyl phosphate (TCP), which is used as additive in hydraulic fluids and engine lubricants. The toxic metabolite 2-(2-cresyl)-4H-1,3,2-benzodioxaphosphorin-2-oxide (CBDP) is formed from the TCP isomer tri-ortho-cresyl phosphate (TOCP) in vivo and is known to react with the active site serine in acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) resulting in the inhibition of the enzymes. Previous in vitro studies showed pronounced species differences in the inhibition kinetics of cholinesterases by organophosphorus compounds (OP), which must be considered in the development of relevant animal models for the investigation of OP poisoning and the aerotoxic syndrome. The present study was designed to investigate the inhibition kinetics of human, Cynomolgus monkey, pig, mini pig, guinea pig, mouse, and rat AChE as well as BChE by CBDP under standardized conditions. There were similar rate constants for the inhibition (ki) of human, Cynomolgus monkey and mouse AChE by CBDP. In contrast, the ki values obtained for guinea pig, mini pig, pig, and rat AChE were 2.8- to 5.9-fold lower than that of human AChE. The results of the present study confirmed CBDP as one of the most potent inhibitors of human BChE, indicating a ki value of 3.24 ± 0.33 ×108M−1min−1, which was about 1,140-fold higher than that of human AChE. Accordingly, a markedly more pronounced inhibition rate of BChE from the species guinea pig, mini pig, pig, rat, Cynomolgus monkey, and mouse by CBDP was found as compared to those of AChE from the respective sources, indicating 2.0- to 89.6-fold higher ki values.