Animal Models Of Human Disease Research Articles

The IL-21/IL-21R signaling axis regulates CD4+ T cell responsiveness to IL-12 to promote bacterial-induced colitis.

IL-21/IL-21R signaling dysregulation is linked to multiple chronic intestinal inflammatory disorders in humans and animal models of human diseases. In addition to its critical requirement for the generation and development of germinal center B cells, IL-21/IL-21R signaling can also regulate the effector functions of a variety of T cell subsets. The antibody-mediated abrogation of IL-21/IL-21R signaling led to the impaired expression of IFN-γ by mucosal CD4+ T cells from human subjects with colitis, suggesting an IL-21/IL-21R-triggered positive feedback loop of the TH1 immune response in the colon. Despite recent advances in our understanding of the mechanisms underpinning the regulation of pro-inflammatory immune responses by the IL-21/IL-21R signaling axis, it remains unclear how this pathway or its downstream molecules contribute to inflammation during bacterial-induced colitis. This study found that IL-21 enhances the surface expression of IL-12Rβ2, but not IL-12Rβ1, in CD4+ T cells, leading to TH1 differentiation and stability. Consistently, these findings also point to an indispensable role of the IL-12Rβ2 signaling axis in promoting pro-inflammatory immune responses during Citrobacter rodentium-induced colitis. Genetic deletion of the IL-12Rβ2 signaling pathway led to the attenuation of C. rodentium-induced colitis in vivo. The genetic deletion of the IL-12Rβ2 signaling pathway did not alter the host's ability to respond adequately to C. rodentium infection or the ability of Il12rb2-/- mice to express antigen-specific cytokines (IFN-γ, IL-17A). IL-21 is a pleiotropic cytokine exerting a wide range of immunomodulatory functions in multiple tissues, and its direct targeting may result in undesirable off-target consequences. These findings highlight the possibility for targeted manipulations of signaling cascades downstream of main regulators of pro-inflammatory responses to control invading pathogens while preserving the integrity of host immune responses. A better understanding of the novel mechanisms by which IL-21/IL-21R signaling regulates bacterial-induced colitis will provide insights into the development of new therapeutic and preventive strategies to harness IL-21/IL-21R signaling or its downstream molecules to treat infectious colitis.

Therapeutic value of homeoprotein signaling pathways.

Cell signaling based on homeoprotein transfer is a pathway with developmental and physiological functions. For a few transcription factors of this family, primarily ENGRAILED1, ENGRAILED2 and OTX2, their physiological functions have led to therapeutic strategies in animal models of human diseases, including Parkinson's disease, amyotrophic lateral sclerosis, amblyopia and anxiety-related disorders. In mesencephalic dopaminergic neurons which degenerate in Parkinson's disease, ENGRAILED1/2 have cell autonomous activities, but their transducing properties enables their use as therapeutic proteins. In contrast, in spinal alpha-motoneurons, which are lost in amyotrophic lateral sclerosis, ENGRAILED1 is supplied by V1 interneurons. Thus, its use as a therapeutic protein to protect alpha-motoneurons against degeneration mimics its normal non-cell autonomous neurotrophic activity. OTX2, synthesized and secreted by the choroid plexus, is transferred to parvalbumin interneurons and exerts regulatory functions controlling cerebral cortex plasticity. Understanding the latter OTX2 function has led to strategies for manipulating visual acuity and anxiety-like behavior in adult mice. In this review, we describe these cases and what is known about the involved molecular mechanisms. Because the transduction sequences are conserved in most of the few hundred homeoproteins, we argue how this family of molecules constitutes an important reservoir of physiological knowledge, with potential consequences in the search for new therapeutic strategies.

A targeted proteomics method for quantifying plasma apolipoprotein kinetics in individual mice using stable isotope labeling

Altered apolipoprotein kinetics play a critical role in promoting dyslipidemia and atherogenesis. Human apolipoprotein kinetics have been extensively evaluated, but similar studies in mice are hampered by the lack of robust methods suitable for the small amounts of blood that can be collected at sequential time points from individual mice. We describe a targeted liquid chromatography tandem mass spectrometry method for simultaneously quantifying the stable isotope enrichment of several apolipoproteins represented by multiple peptides in serial blood samples (15 μl each) obtained after retro-orbital injection of 13C6,15N2-lysine (Lys8) in mice. We determined apolipoprotein fractional clearance rates (FCRs) and production rates (PRs) in WT mice and in two genetic models widely used for atherosclerosis research, LDL receptor–deficient (Ldlr−/−) and apolipoprotein E–deficient (Apoe−/−) mice. Injection of Lys8 produced a unique and readily detectable mass shift of labeled compared with unlabeled peptides with sensitivity allowing robust kinetics analyses. Ldlr−/− mice showed slower FCRs of APOA1, APOA4, total APOB, APOB100, APOCs, APOE and APOM, while FCRs of APOA1, APOB100, APOC2, APOC3, and APOM were not lower in Apoe−/− mice versus WT mice. APOE PR was increased in Ldlr−/− mice, and APOB100 and APOA4 PRs were reduced in Apoe−/− mice. Thus, our method reproducibly quantifies plasma apolipoprotein kinetics in different mouse models. The method can easily be expanded to include a wide range of proteins in the same biospecimen and should be useful for determining the kinetics of apolipoproteins in animal models of human disease.

Two-photon excitation fluorescence in ophthalmology: safety and improved imaging for functional diagnostics.

Two-photon excitation fluorescence (TPEF) is emerging as a powerful imaging technique with superior penetration power in scattering media, allowing for functional imaging of biological tissues at a subcellular level. TPEF is commonly used in cancer diagnostics, as it enables the direct observation of metabolism within living cells. The technique is now widely used in various medical fields, including ophthalmology. The eye is a complex and delicate organ with multiple layers of different cell types and tissues. Although this structure is ideal for visual perception, it generates aberrations in TPEF eye imaging. However, adaptive optics can now compensate for these aberrations, allowing for improved imaging of the eyes of animal models for human diseases. The eye is naturally built to filter out harmful wavelengths, but these wavelengths can be mimicked and thereby utilized in diagnostics via two-photon (2Ph) excitation. Recent advances in laser-source manufacturing have made it possible to minimize the exposure of in vivo measurements within safety, while achieving sufficient signals to detect for functional images, making TPEF a viable option for human application. This review explores recent advances in wavefront-distortion correction in animal models and the safety of use of TPEF on human subjects, both of which make TPEF a potentially powerful tool for ophthalmological diagnostics.

Central Role of β-1,4-GalT-V in Cancer Signaling, Inflammation, and Other Disease-Centric Pathways.

UDP-Galactose: Glucosylceramide, β-1,4-Galactose transferase-V (β-1,4-GalT-V), is a member of a large glycosyltransferase family, primarily involved in the transfer of sugar residues from nucleotide sugars, such as galactose, glucose mannose, etc., to sugar constituents of glycosphingolipids and glycoproteins. For example, UDP-Galactose: Glucosylceramide, β-1,4-galactosyltransferase (β-1,4-GalT-V), transfers galactose to glucosylceramide to generate Lactosylceramide (LacCer), a bioactive "lipid second messenger" that can activate nicotinamide adenine dinucleotide phosphate(NADPH) oxidase (NOX-1) to produce superoxide's (O2-) to activate several signaling pathways critical in regulating multiple phenotypes implicated in health and diseases. LacCer can also activate cytosolic phospholipase A-2 to produce eicosanoids and prostaglandins to induce inflammatory pathways. However, the lack of regulation of β-1,4-GalT-V contributes to critical phenotypes central to cancer and cardiovascular diseases, e.g., cell proliferation, migration, angiogenesis, phagocytosis, and apoptosis. Additionally, inflammation that accompanies β-1,4-GalT-V dysregulation accelerates the initiation and progression of cancer, cardiovascular diseases, as well as inflammation-centric diseases, like lupus erythematosus, chronic obstructive pulmonary disease (COPD), and inflammatory bowel diseases. An exciting development in this field of research arrived due to the recognition that the activation of β-1,4-GalT-V is a "pivotal" point of convergence for multiple signaling pathways initiated by physiologically relevant molecules, e.g., growth factors, oxidized-low density lipoprotein(ox- LDL), pro-inflammatory molecules, oxidative and sheer stress, diet, and cigarette smoking. Thus, dysregulation of these pathways may well contribute to cancer, heart disease, skin diseases, and several inflammation-centric diseases in experimental animal models of human diseases and in humans. These observations have been described under post-transcriptional modifications of β-1,4- GalT-V. On the other hand, we also point to the important role of β-1-4 GalT-V-mediated glycosylation in altering the formation of glycosylated precursor forms of proteins and their activation, e.g., β-1 integrin, wingless-related integration site (Wnt)/-β catenin, Frizzled-1, and Notch1. Such alterations in glycosylation may influence cell differentiation, angiogenesis, diminished basement membrane architecture, tissue remodeling, infiltrative growth, and metastasis in human colorectal cancers and breast cancer stem cells. We also discuss Online Mendelian Inheritance in Man (OMIM), which is a comprehensive database of human genes and genetic disorders used to provide information on the genetic basis of inherited diseases and traits and information about the molecular pathways and biological processes that underlie human physiology. We describe cancer genes interacting with the β-1,4-GalT-V gene and homologs generated by OMIM. In sum, we propose that β-1,4-GalT-V gene/protein serves as a "gateway" regulating several signal transduction pathways in oxidative stress and inflammation leading to cancer and other diseases, thus rationalizing further studies to better understand the genetic regulation and interaction of β-1,4-GalT-V with other genes. Novel therapies will hinge on biochemical analysis and characterization of β-1,4-GalT-V in patient-derived materials and animal models. And using β-1,4-GalT-V as a "bonafide drug target" to mitigate these diseases.

Regulation of Immune Checkpoint Antigen CD276 (B7-H3) on Human Placenta-Derived Mesenchymal Stromal Cells in GMP-Compliant Cell Culture Media.

Therapies utilizing autologous mesenchymal cell delivery are being investigated as anti-inflammatory and regenerative treatments for a broad spectrum of age-related diseases, as well as various chronic and acute pathological conditions. Easily available allogeneic full-term human placenta mesenchymal stromal cells (pMSCs) were used as a potential pro-regenerative, cell-based therapy in degenerative diseases, which could be applied also to elderly individuals. To explore the potential of allogeneic pMSCs transplantation for pro-regenerative applications, such cells were isolated from five different term-placentas, obtained from the dissected maternal, endometrial (mpMSCs), and fetal chorion tissues (fpMSCs), respectively. The proliferation rate of the cells in the culture, as well as their shape, in vitro differentiation potential, and the expression of mesenchymal lineage and stem cell markers, were investigated. Moreover, we studied the expression of immune checkpoint antigen CD276 as a possible modulation of the rejection of transplanted non-HLA-matched homologous or even xeno-transplanted pMSCs. The expression of the cell surface markers was also explored in parallel in the cryosections of the relevant intact placenta tissue samples. The expansion of pMSCs in a clinical-grade medium complemented with 5% human platelet lysate and 5% human serum induced a significant expression of CD276 when compared to mpMSCs expanded in a commercial medium. We suggest that the expansion of mpMSCs, especially in a medium containing platelet lysate, elevated the expression of the immune-regulatory cell surface marker CD276. This may contribute to the immune tolerance towards allogeneic pMSC transplantations in clinical situations and even in xenogenic animal models of human diseases. The endurance of the injected comparably young human-term pMSCs may promote prolonged effects in clinical applications employing non-HLA-matched allogeneic cell therapy for various degenerative disorders, especially in aged adults.

45 Genome Editing of Livestock for Production Agriculture and Biomedical Applications

Abstract Continued population growth and clear evidence of climate change are placing unprecedented strain on food production and health care systems worldwide. Agricultural production goals that highlight environmental sustainability are at the forefront of social dialog, with animal agriculture routinely targeted for inefficiencies, welfare concerns and impact on climate. Genomic selection has transformed animal production and the rate of genetic gain in each generation continues to increase. Nonetheless, it is apparent that the rate of production gains will fall short of the necessary goals of increased efficiency and adaptation to the changing environment. Genetic engineering may have an important role in enhancing livestock production as well as providing new models for research in human health. The CRISPR/Cas and related systems for targeted gene editing has revolutionized livestock genetic engineering by increasing the rate of targeted mutation, thus drastically reducing the cost of producing founder animals with the desired trait. Intentional insertion of well-known mutations into genes such as myostatin have been shown to impart the expected increase in meat production, but have also resulted in undesirable associated traits affecting fertility, marbling and the skeleton. Thus, the approach to increasing meat production may require nuances such as promotor targeting and multiple gene targeting to obtain the desired production characteristics. Efforts to increase disease resistance could have dramatic impacts on animal welfare and production rates, potentially saving millions of animals from morbidity and mortality. However, confronting a single disease with each genetic alteration will be costly and inefficient. Considerations on how broad scale disease resistance could be achieved through genetic engineering will be instrumental in developing livestock with advantages to avoid infection and/or transmission of multiple pathogens. Gene editing also allows correction of mutations that arise in livestock through natural mechanisms while under intense genomic selection for increased production. Removing disease causing mutations in otherwise high value livestock will allow continued production from these lines without concern for inheriting the causative mutation. The same process can be utilized to insert genetic mutations known to phenocopy human conditions providing valuable large animal models of human genetic disease. Increased milk, meat, or eggs per animal unit will sustainably improve high quality, nutrient dense food production while large animal models of human disease will provide a more translational biomedical model for accelerated development of drugs and treatments. These goals are achievable, but require continued investment in innovation, clear regulatory framework for commercialization, education, and public engagement to ensure success.

Animal Models of Human Disease

The use of animal models of human disease is critical for furthering our understanding of disease mechanisms, for the discovery of novel targets for treatment, and for translational research. This Special Topic entitled “Animal Models of Human Disease” aimed to collect state-of-the-art primary research studies and review articles from international experts and leading groups using animal models to study human diseases. Submissions were welcomed on a wide range of animal models and pathologies, including infectious disease, acute injury, regeneration, cancer, autoimmunity, degenerative and chronic disease. Seven participating MDPI journals supported the Special Topic, namely: Biomedicines, Cells, Current Issues in Molecular Biology, Diagnostics, Genes, the International Journal of Molecular Sciences, and the International Journal of Translational Medicine. In total, 46 papers were published in this Special Topic, with 37 full length original research papers, 2 research communications and 7 reviews. These contributions cover a wide range of clinically relevant, translatable, and comparative animal models, as well as furthering understanding of fundamental sciences, covering topics on physiological processes, on degenerative, inflammatory, infectious, autoimmune, neurological, metabolic, heamatological, hormonal and mitochondrial disorders, developmental processes and diseases, cardiology, cancer, trauma, stress, and ageing.

TRPM channels in health and disease.

Different cell channels and transporters tightly regulate cytoplasmic levels and the intraorganelle distribution of cations. Perturbations in these processes lead to human diseases that are frequently associated with kidney impairment. The family of melastatin-related transient receptor potential (TRPM) channels, which has eight members in mammals (TRPM1-TRPM8), includes ion channels that are highly permeable to divalent cations, such as Ca2+, Mg2+ and Zn2+ (TRPM1, TRPM3, TRPM6 and TRPM7), non-selective cation channels (TRPM2 and TRPM8) and monovalent cation-selective channels (TRPM4 and TRPM5). Three family members contain an enzymatic protein moiety: TRPM6 and TRPM7 are fused to α-kinase domains, whereas TRPM2 is linked to an ADP-ribose-binding NUDT9 homology domain. TRPM channels also function as crucial cellular sensors involved in many physiological processes, including mineral homeostasis, blood pressure, cardiac rhythm and immunity, as well as photoreception, taste reception and thermoreception. TRPM channels are abundantly expressed in the kidney. Mutations in TRPM genes cause several inherited human diseases, and preclinical studies in animal models of human disease have highlighted TRPM channels as promising new therapeutic targets. Here, we provide an overview of this rapidly evolving research area and delineate the emerging role of TRPM channels in kidney pathophysiology.

A Green and Cost-Effective Chromatographic Method for the Measurement of the Activity of Human Serum Carnosinase

Carnosinase (i.e., CN1; E.C. 3.4.13.20) is an enzyme found in the sera of higher primates. CN1 preferentially catalyzes the hydrolysis of natural, orally adsorbed histidine dipeptides like carnosine (i.e., β-alanyl-L-histidine). This is the reason why carnosine has a limited use as a human food supplement or pharmacological agent, despite the promising results obtained in experiments on animal models of human diseases. Herein, an assay is reported for the measurement of serum CN1 activity. The method is intended for the screening of CN1 inhibitors able to enhance carnosine bioavailability in humans. The method was developed to monitor serum hydrolytic activity via the quantitation of one of the products of carnosine hydrolysis (i.e., histidine). Separation was achieved without using organic solvents by means of ion chromatography (IC), whereas detection was provided by UV spectroscopy. The assay herein reported is a green and cost-effective alternative to a recently published method based on hydrophilic interaction liquid chromatography (HILIC) and mass spectrometry (MS). The results show that such a method produces reliable measurements of serum hydrolytic activity and can be used for the screening of CN1 inhibitors.

Recent developments in targets for ischemic foot disease.

Diabetes is a key risk factor for ischaemic foot disease, which causes pain, tissue loss, hospital admission, and major amputation. Currently, treatment focuses on revascularisation, but many patients are unsuitable for surgery and revascularisation is frequently unsuccessful. The authors describe recent research in animal models and clinical trials investigating novel medical targets for ischaemia, including theories about impaired wound healing, animal models for limb ischaemia and recent randomised controlled trials testing novel medical therapies. Novel targets identified in animal models included stimulating mobilisation of CD34+ progenitor cells through upregulating oncostatin M or microRNA-181, downregulating tumour necrosis factor superfamily member 14, or activating the Wingless pathway. Within the ischaemic limb vasculature, upregulation of apolipoprotein L domain containing 1, microRNA-130b or long noncoding RNA that enhances endothelial nitric oxide synthase expression promoted limb blood supply recovery, angiogenesis, and arteriogenesis. Similarly, administration of soluble guanylate cyclase stimulators riociguat or praliciguat or 3-ketoacyl-CoA thiolase inhibitor trimetazidine promoted blood flow recovery. Translating pre-clinical findings to patients has been challenging, mainly due to limitations in clinically translatable animal models of human disease. Promising results have been reported for administering plasmids encoding hepatocyte growth factor or intra-arterial injection of bone marrow derived cells in small clinical trials. It remains to be seen whether these high resource therapies can be developed to be widely applicable. In conclusion, an ever-expanding list of potential targets for medical revascularisation is being identified. It is hoped that through ongoing research and further larger clinical trials, these will translate into new broadly applicable therapies to improve outcomes.

Enrichment analysis of phenotypic data for drug repurposing in rare diseases.

Drug-induced Behavioral Signature Analysis (DBSA), is a machine learning (ML) method for in silico screening of compounds, inspired by analytical methods quantifying gene enrichment in genomic analyses. When applied to behavioral data it can identify drugs that can potentially reverse in vivo behavioral symptoms in animal models of human disease and suggest new hypotheses for drug discovery and repurposing. We present a proof-of-concept study aiming to assess Drug-induced Behavioral Signature Analysis (DBSA) as a systematic approach for drug discovery for rare disorders. We applied Drug-induced Behavioral Signature Analysis to high-content behavioral data obtained with SmartCube®, an automated in vivo phenotyping platform. The therapeutic potential of several dozen approved drugs was assessed for phenotypic reversal of the behavioral profile of a Huntington's Disease (HD) murine model, the Q175 heterozygous knock-in mice. The in silico Drug-induced Behavioral Signature Analysis predictions were enriched for drugs known to be effective in the symptomatic treatment of Huntington's Disease, including bupropion, modafinil, methylphenidate, and several SSRIs, as well as the atypical antidepressant tianeptine. To validate the method, we tested acute and chronic effects of tianeptine (20mg/kg, i. p.) in vivo, using Q175 mice and wild type controls. In both experiments, tianeptine significantly rescued the behavioral phenotype assessed with the SmartCube® platform. Our target-agnostic method thus showed promise for identification of symptomatic relief treatments for rare disorders, providing an alternative method for hypothesis generation and drug discovery for disorders with huge disease burden and unmet medical needs.

The Use of Zebrafish and in Vivo PAL as a Novel Discovery Platform for Psychoactive Agents

Our research is utilizing the zebrafish model to study molecular processes and animal models of human diseases. These studies discussed here start to assess how different bath salts and methamphetaminecause death upon overdose by addressing the molecules that may bind to the illicit drugs. We have accomplished this by developing an in vivo photo-affinity labelling (PAL) system and studying the drugs in our system. The zebrafish has been lauded as a unique and powerful tool to examine development in a vertebrate animal and address molecular aspects of human disease for decades. We will examine recent published data from our laboratory utilizing the PAL system we developed.

CRISPR/Cas Systems for Therapeutic Purposes: Applications in Disease Prevention, Mechanism Study, and Treatment

CRISPR/Cas systems were novel multifunctional genome editing techniques, and the modification of Cas proteins endowed them with more efficient and precise properties. Herein, we systematically outlined the principles and applications of CRISPR/Cas systems in disease prevention, mechanism research, and gene treatment of diseases. Specifically, we detailed the systems widely utilized, containing the Cas9, Cas12a, and Cas13a systems, and summarized the current status of the medical applications of these systems in terms of vaccine design, trait improvement, animal models of human diseases, organ transplantation, gene therapy, and gene detection. We concluded that CRISPR/Cas systems had great potential to advance medical research and practice but also should be carefully regulated and supervised to ensure their safety and efficacy.

The International Society for Biological and Environmental Repositories Presents Abstracts from Its Annual Meeting

Biopreservation and BiobankingAhead of Print AbstractsFree AccessThe International Society for Biological and Environmental Repositories Presents Abstracts from Its Annual Meeting“Come As You Are: Building Biobanking Bridges” In‐Person: May 3–6, 2023 Seattle, Washington Virtual: June 6–7, 2023Published Online:14 Apr 2023https://doi.org/10.1089/bio.2023.29118.abstractsAboutSectionsPDF/EPUB Permissions & CitationsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail The abstracts that follow demonstrate the broad range of timely issues addressed in the contributed Oral and Poster presentations at ISBER's 2023 Annual Meeting & Exhibits.Oral AbstractO‐01 Accelerating Translational Research by Enabling Access to Specimens and DatasetsS. Joshi1, E. Matzke2, D. Galipeau31Fred Hutchinson Cancer Research Center, Seattle, Washington, United States, 2Provincial Health Services Authority, Vancouver, British Columbia, Canada, 3Oregon Health & Science University, Portland, Oregon, United StatesStatement of the Problem: Finding appropriate clinical specimens and associated data often causes bottlenecks in the translational research pipeline. Often investigators procure specimens through only one or a small number of sources which often have limited volume of participants and demographics of the patient population. Seemingly paradoxical, groups with large numbers of banked specimens often struggle to increase awareness and use of their collected specimens, which can lead to limited return on investment for high infrastructure costs.Proposed Solution: To address the above‐mentioned challenges the Specimen & Data Acquisition Network (SAN) was formed in 2019 to increase opportunities for resource exchange in the Pacific Northwest region (Oregon, Washington, and British Columbia). The concept of SAN, a hub and spoke network to speed sharing of resources and ideas, originated in response to sparse specimen resources within our translational research program which was a key bottleneck to research endeavors. Building on existing local connections and those from our research network in other academic institutions, we began collaborations with 7 additional institutions that were aligned with our objectives and goals. SAN serves as a liaison for investigators to leverage collective resources and works to support researchers in their biobanking activities. SAN is a conduit for collaborative science across the regional translational research community.Conclusions: Currently, the SAN consists of 10 member organizations, each with representation on the governance committee. SAN began serving investigators in 2020 and, to date, has received 80 requests and connected more than 40 investigators to critical project resources spanning 12 institutions. SAN works with researchers to provide support across 6 service areas: 1) Access to specimens, 2) Access to datasets, 3) Data Management Support, 4) Research Connections, 5) Regulatory Guidance, and 6) Resource Lifecycle Management. These supportive services are catered to give researchers more time to focus on their next big discovery. Five SAN member institutions are signatory to an umbrella material/data transfer agreement that has expedited resource sharing. We envision that this will be a valuable resource for the translational research community.O‐02 Power of Consortium Shared Resources When External Threats OccurS. Hume4, 1, S. Higgins1, 2, A. J. Mountain1, 3, C. Gilfillan4, W. Ng1, 21Victorian Cancer Biobank Consortium, Melbourne, Victoria, Australia, 2Cancer Council Victoria, Melbourne, Victoria, Australia, 3Austin Health Tissue Bank, Austin Health, Heidelberg, Victoria, Australia, 4Eastern Health Tissue Bank, Eastern Health, Box Hill, Victoria, AustraliaStatement of Problem: The Eastern Health Tissue Bank (EHTB) is a hospital‐integrated cancer biobank and a member of Victorian Cancer Biobank (VCB) Consortium. In 2021, the biobank responded to several external threats that affected daily operations. These included a cyber‐threat incident at the hosting institution and multiple COVID‐19‐related disruptions as the city experienced one of the longest pandemic lockdowns globally.Unplanned events can severely and suddenly restrict biobanking activities, and small biobanks require support to ensure sustainability and business continuity.Proposed Solution: As part of the VCB Consortium, the EHTB has a strong resilience and ability to respond to external threats or incidents. As a hub‐and‐spoke Consortium (comprising a central lead agency with five cancer tissue banks), the VCB utilises shared resources to strengthen business continuity.This presentation depicts two scenarios (cyber threat and pandemic) which demonstrate the combined use of tools across a networked biobank to reduce risks and impacts from large‐scale organisational disruptions.Three Consortium‐wide tools were utilised:1) A robust Business Continuity Plan, incorporating key stakeholders' responsibilities in risk management.2) A Consortium Business Plan with local (institution) and networked (consortium) goals.3) Implementation of harmonised Standardised Operating Procedures (SOPs).Scenario Outcomes Included:1) Central Operations “Hub” provided immediate contingency support as part of a framework for rapid response/recovery, including facilitating communications with stakeholders.2) A partner “Spoke” Site allowed EHTB staff access to Biobank shared databases, group business communication platforms, and shared biospecimen storage.3) Harmonised Consortium SOPs allowed large‐scale multisite dispatches to still be completed.Conclusions: In responding to external threat incidents, a robust business continuity plan and immediate access to shared resources enabled the EHTB to retain some functionality. If the EHTB had been a standalone biobank, capacity to operate would have been diminished, resulting in the loss or suspension of activity.The resources described cannot be underestimated in their effectiveness when an external threat occurs.O‐03 The Qualification in Biorepository Science (QBRS) Examination: Progress Report on its Global ReachB. Schacter1, 2, D. Simeon‐Dubach31Medical Oncology and Hematology, CancerCare Manitoba, Winnipeg, Manitoba, Canada, 2Internal Medicine, University of Manitoba, Winnipeg, Manitoba, Canada, 3medservice Daniel Simeon Dubach, Walchwil, ZG, SwitzerlandBackground: The importance of quality parameters in biobanking of biological materials like human tissues has been emphasized in the past: Biobanking standards by ISO (ISO 20387), CAP accreditation, Best Practices (ISBER Best Practices 4th ed.), OECD standards, and Standard Operating Practices and Policies by CTRNet were developed to address these challenges.While these procedures are proving invaluable to biobanks to ensure that their processes for accessing, processing, and distributing biobanked materials result in reproducibility and quality performance, they do not address the question of quality performance in the biobanking professionals themselves.Methods: The International Society for Biological and Environmental Repositories (ISBER) and ASCP BOC (American Society for Clinical Pathology Board of Certification) as two leading organisations in their fields joined forces to develop the QBRS (Qualification in Biorepository Science) examination, in order to further enhance reproducible and sustainable quality biobanking in biobanking professionals. The exam is administered online and is unique in its field.Well trained and skilled staff are essential to ensure high quality biospecimens for reproducible results in precision medicine, biomarker development, and biomedical research as a whole, while aiming for sustainable biobanking activities.Results: To date, over 58 individuals in Australia, Hong Kong, Singapore, Ireland, Wales, Nigeria, Saudi Arabia, UAE, Costa Rica, and the USA have taken and passed the exam to qualify for the QBRS, demonstrating its global appeal.Conclusions: The QBRS has become the standard which defines the required level of knowledge and skill in biobanking professionals that will assure sustained quality and reproducibility in biobanking laboratories worldwide. Successful applicants have acknowledged that achieving the QBRS makes them more desirable in hiring competitions and enhances their career trajectory. Conversely, biobanks that hire a QBRS qualified biobanker have acknowledged that they are striving for quality in their biobanking operation. The QBRS qualification establishes that the biobank in which the qualified biobanker works is one that promotes and establishes quality within its biobanking staff. Biobank directors have indicated that the QBRS exam provides further impetus for professional training for their staff.O‐04 A New Tool to Match Organ, Tissue, and Eye Post‐mortem Donors to Research Project CriteriaK. D. Kunkle, G. H. Grossman, T. Cattell, A. AbbottPrecision Ocular Biobank, Advancing Sight Network, Birmingham, Alabama, United StatesBackground: Post‐mortem repositories recover organs, tissues, and eyes for research from authorized donors. However, due to the rapid nature of the donation process conflicting with the laborious task of matching donors to research project criteria, organizations miss opportunities to identify donors for prospective recovery. The ReSync app was developed by the Precision Ocular Biobank to assist in the real‐time identification of research donors at the initial point of the referral call.Methods: ReSync is an app built in Claris FileMaker 19, a low‐code relational database engine. Although ReSync is a closed system, open‐source functionality of Claris FileMaker allows it to universally connect to external donation referral databases. Research project criteria are entered into a table in ReSync prior to activation. Exclusion and inclusion criteria in categories such as donor demographics, medical history, and referral status act as triggers to match with donor information. Once active, referral coordinators enter donor information into the host database during the initial call from the hospital with a recent or impending death. Filtering occurs through connecting pairs of fields in tables in ReSync and the host database to validate or invalidate a donor candidate against each project. When a match between a project and donor occurs, a flag alerts the coordinator in real‐time. Matching projects appear in a pop‐up window, and referral coordinators can then consult with research specialists or directly assign the case to one of the projects and initiate recovery dispatch.Results: Since its employment, ReSync has allowed the Precision Ocular Biobank to expand its open research requests from 3 to 18 at any given point, representing a six‐fold increase in concomitant projects. This also led to a dramatically decrease in the time a donor was matched to a project, from twenty minutes to five (400% decrease). Even with more projects open simultaneously, request fulfillment time decreased by 25%.Conclusions: Low‐code, automated donor matching solutions allow post‐mortem repositories with limited software developing capability to better fulfil prospective research requests. While simultaneously reducing missed opportunities and eligibility times, programs like ReSync allow for the expansion of enrolled projects.O‐05 Determining Pre‐analytical Variables and Molecular Determinants to Ensure High Quality of Luxor BiobankN. Kordy, S. Ezzat, S. Mahmoud, R. Mohamed, A. Hagag, A. M. Gamal, M. M. Saady, A. SalehResearch Department, Shefa Al Orman Hospital, Luxor, EgyptBackground: Biobank sample preparation methods must ensure long‐term functionality of the products obtained. Preanalytical variables affect the integrity of the biospecimens, and results of analyses. The quality of nucleic acids has a major importance for molecular biology techniques used in genetic analysis. Quality indicators are the tool to measure these parameters; purity and integrity are the determinants for DNA quality. This study aims to determine the quality of SOH Biobank (SOHB) by independently evaluating the preanalytical variables in terms of measurement of processing time, aliquots position audit, and the quality of DNA samples.Methodology: SOHB biospecimen collection is performed after SOH‐IRB approval. Standard operating procedures (SOPs) were written with strict adherence by the team. Blood samples are collected after participant recruitment in the biobank program and obtaining informed consent. After blood collection, the blood is processed within one‐hour and stored in ‐80 degree Celsius freezers. Quality control was determined first, through recording sample withdrawal, receiving, and processing time. Second, aliquots position audits were performed manually on recruited patients for the last 6 months of 2021 to ensure 100% accurate aliquots position, Finally, DNA extracted from buffy coat samples was stored in the duration of 01‐2017 to 02‐2022 using QIAamp DNA kit (Qiagen). Concentration and purity are measured by spectrophotometer (Multisky scan, Thermofisher). Pure DNA ratio should be between 1.8 to 2.0; a higher ratio reveals DNA degradation, while a lower ratio indicates protein contamination.Results: Mean value of processing time for 6600 samples is calculated to ensure that samples are processed and stored in ‐80 degree Celsius freezers within 1 hour of sample withdrawal. Results showed the mean value is 23 ± 0.08 minutes. Manual aliquots position audit was performed for samples stored from July to December 2021; 1087 cases showed 96% accuracy, while 4% aliquot misassigned position inside box. Purity detection for 980 samples out of 6600 samples were analyzed and showed a mean of A260/A280ratio of 1.8 ± 0.025.Conclusion: SOHB quality control results are satisfactory ensuring that SOHB can provide high‐quality samples for genomic studies. Quality control should be performed in a regular manner to ensure the appropriate management of the samples, avoiding low confidence in results, high costs, and sample wasting.O‐06 MIRRI‐ERIC: A Pan‐European Research Infrastructure for Making Microbial Science and Innovation HappenR. Aznar1, M. Bosschaerts2, N. Lima3, L. Soares41Departamento de Microbiología y Ecología and Colección Española de Cultivos Tipo (CECT), Universitat de València, Valencia, Spain, 2BCCM Coordination Cell, Belgian Science Policy, Brussels, Belgium, 3CEB‐Universidade do Minho, Micoteca da Universidade do Minho, Braga, Portugal, 4MIRRI‐ERIC Central Coordinating Unit, MIRRI‐ERIC, Braga, PortugalStatement of the Problem: Several hurdles hamper microbial research and cause inefficient, ineffective, and costly science and innovation: difficulty to access strains cited in papers and databases; sharing of unverified strains; gaps in the current offer; fragmentation of resources, data, services, and expertise; and deficient support to individual culture collections (CCs).Proposed Solution: The Microbial Resource Research Infrastructure ‐ European Research Infrastructure Consortium (MIRRI‐ERIC) has been created as a pan‐European distributed research infrastructure, with the goal of solving/mitigating these bottlenecks, promoting complementarity, reducing redundancy, and continuously improving the capacity of its Partner Microbial domain Biological Resource Centres (mBRCs). MIRRI brings together ∼50 mBRCs, CCs, and research institutes from ten European countries working for the preservation, systematic investigation, provision, and valorisation of microbial resources and biodiversity, with a special focus on the domains of Health & Food, Agro‐Food, and Environment & Energy.MIRRI serves the bioscience and the bioindustry communities by facilitating the access, through a single point, to a catalogue of 400,000+ high‐quality microbial resources, such as bacteria, cyanobacteria, archaea, yeasts, filamentous fungi, micro‐algae, bacteriophages, viruses, and other microbiological material, such as microbiomes, plasmids, and genomic DNA. The MIRRI Information System will also provide users with resources' associated data, as available – e.g., taxonomy, ecology, pathogenicity, morphology, physiology, chemical characterization, DNA barcoding, or genomics.Based on its partner organisations' state‐of‐the‐art facilities/equipment and top‐level expertise, MIRRI also offers its users a diverse catalogue of high‐quality services, including pipelines of integrated, product‐oriented services made available as tailor‐made, turnkey solutions.Conclusions: The resources, services, and expertise provided by MIRRI and its partners can help researchers and bioindustries deliver the maximum value and impacts from their projects, technologies, and products.MIRRI is continuously engaged in enlarging its coverage in Europe and beyond. EU Member States, associated countries, third countries other than associated countries, and intergovernmental organisations may become a Member or an Observer of MIRRI‐ERIC. Individual organisations may become Partners of MIRRI‐ERIC.O‐07 Developing Genomic and Biomaterials Resources for Establishing Nonhuman Primate Models of Human DiseaseE. J. Vallender1, 2, S. M. Peterson3, A. Lewis3, A. J. Ericsen4, 5, K. G. Ray3, B. N. Bimber3, B. Ferguson31The University of Mississippi Medical Center, Jackson, Mississippi, United States, 2Tulane National Primate Research Center, Covington, Louisiana, United States, 3Oregon Health & Science University Oregon National Primate Research Center, Beaverton, Oregon, United States, 4Emory University, Atlanta, Georgia, United States, 5Emory National Primate Research Center, Atlanta, Georgia, United StatesBackground: Animal models of human disease play a vital role in biomedical research and therapeutic development. Nonhuman primates, in particular rhesus macaques, share behavioral, anatomical, physiological, and genetic similarities with humans that make them excellent translational models. The wealth of information developed on these animals through diverse research efforts can be leveraged through interconnected repositories of phenotypic data, biomaterials, and genomic information. Through the development of a centralized framework it is possible to accelerate the generation of nonhuman primate models of human disease and effect vital synergies that go beyond individual laboratories or research programs.Methods: The macaque Genotype and Phenotype (mGAP) resource serves as a repository for genomic data derived from rhesus macaques, largely those found at NIH‐funded Primate Research Centers. A standardized bioinformatic pipeline is used to call single nucleotide variation (SNV) and to cross reference human data on regulatory elements, disease‐associated variation, and evolutionary conservation. SNVs are also annotated using variant effect prediction tools. Data are made openly available to the research community both in aggregate as well as at the individual animal level. This information can then be tied back to specific animal health records, phenotypic information, and biomaterials through the contributing centers.Results: mGAP currently houses genomic information from 2,985 rhesus macaques including nearly 52 million variants. Of these, 11.5 thousand overlap with human ClinVar annotations and 33.2 thousand overlap with GWAS associations housed in GRASP. Many more, 125 thousand, are predicted deleterious by Polyphen2 or high impact by SnpEff. Coupled with phenotypic information derived from animal health records and pathology reports, spontaneously occurring genetic models of multiple rare human diseases have been identified and are being used for novel therapeutic development and biomarker discovery.Conclusions: Through the integration of genomic and phenotypic information, rhesus macaques are being developed and harnessed for translationally important nonhuman primate animal models. These efforts can occur at scale only through resources that bring together diverse data and research expertise while still allowing for the connection to individual animals and samples.O‐08 AI in Medicine Based on Biobank Data ‐ New Opportunities and (Not) Known RisksK. SargsyanInternational Biobanking and Education, Medical University of Graz, Graz, STMK, AustriaThe possibilities for improving effectiveness and interoperability clearly show the directions in which BBs will develop soon. Today, virtual links between different DAs allow an exchange of data and samples, which previously could only occur with great effort and long waiting times or even rejections of queries, have decreased. Furthermore, the notion of a biological “sample” is extended to imaging samples, which multiplies the amount and dimensionality of the data obtained and thus provides biobanks with a more complete picture of individual patients and entire disease patterns. With the integration of digitalization into biobanks, a steady flow of data is ensured that, in the best case, does not cease for the duration of a patient's lifetime. This steady flow of data updates and exchange enables better tracking of the health status of individual patients and populations and can provide far‐reaching and as yet unknown information for the epidemiology of certain diseases in terms of course, treatment response, and prognosis. Particularly concerning genotype‐phenotype predictions, there is the potential to use genetic traits to develop new personalized therapies or targeted therapies and to identify subgroups that, for example, have an increased risk of developing certain diseases based on their genetic features or to facilitate the early detection of the respective diseases and, among other things, to single out non‐responders and to focus more specifically on their specific needs. The consent forms that patients and donors have to fill out and accept with their signature and consent when providing samples to clinical institutes or biobanks, or similar repositories will also evolve into more flexible, dynamic, digitally accessible forms that enable patients/donors to track their own samples and their informative value or usefulness in research whenever and wherever they are. They can withdraw their consent at any time and thus exercise complete control over their genetic material processing. Thus, they will be better treated, jet better informed, and educated.O‐09 Biobanking during War: Experience of Audubon BioscienceM. Yanovytska, B. Shkurupii, H. Chytaieva, E. Serdyukova, A. Shekhovtsov, I. Voievoda, A. Yudchenko, N. Sobetska‐Koloda, T. Nguyen, A. Giardina, A. Popova, R. SemikovAudubon Bioscience, Kyiv, UkraineOn February 24, 2022, the large‐scale Russian army invasion severely interrupted the operations of our biobank in Ukraine. Our team globally and especially in Ukraine faced and is still facing many challenges. Before the events occurred (when risks of the war escalation increased), we prepared our contingency and business continuity plans. They were partially executed. We have since been able to resume operations in Ukraine.On the first day of the large‐scale war, the Operations team, together with Revenue and HR teams, organized the evacuation of the staff with all documentation and laptops, as well as laboratory equipment and ambient storage specimens to the Western region of Ukraine. We were able to eventually move the ambient specimens to the US. Once in our US operations facility, we reorganized and cataloged every specimen and reestablished our biobank there. For frozen specimens, since we can't relocate them due to the lack of proper transportation at that time, we decided to seal all freezers with specimens and installed surveillance cameras to keep them safe. Together with our temperature monitoring system, we were able to keep track of our freezers' temperature and ensured that the specimens were properly stored during the 2‐month period when only a few members of our team remained at the biobank.Ongoing Ukrainian projects were immediately transferred to other countries of operations such as Turkey, USA, Armenia, etc. In three months (May 2022) we achieved financial break‐even (monthly) confirming the fact that global organization was stabilized. After two months, when active warfare was stopped in the Kyiv region, new logistic chains were organized and we started to rebuild our workflow. By August 2022, the Ukrainian operations reached the level of pre‐war collection rates. However, we continue to distribute projects among all of our affiliates in other countries to minimize risks for our partners.In conclusion, the operations of Ukrainian biobanking have been faced with severe difficulties due to the Russian‐Ukrainian war. Although we were not able to execute our contingency and business continuity plans in full, right planning together with good execution and improvisation helped minimize the negative effect. Solutions that we put in place to ensure proper specimen storage were implemented and proved to be effective. Our ability to redistribute projects globally enabled us to continue supporting precision medicine researchers.O‐10 Survey of Adolescents Regarding Their Opinion of Research and Vaccination During the COVID‐19 PandemicQ. Aujla1, I. Kayda1, A. Ellis1, D. Goldfarb1, 2, J. Bettinger2, 3, L. Mâsse2, 3, J. Bush1, 2, S. Vercauteren1, 21Pathology and Lab Medicine, BC Children's Hospital, Vancouver, British Columbia, Canada, 2Faculty of Medicine, The University of British Columbia, Vancouver, British Columbia, Canada, 3BC Children's Hospital Research Institute, Vancouver, British Columbia, CanadaBackground: The COVID‐19 pandemic has emphasized the importance of research into disease biology, prevention, and treatment of emerging diseases and pathogens. Currently, there is limited understanding on adolescent perceptions towards biobanking, research participation, vaccinations, and how the pandemic has altered these perceptions. The BC Children's Hospital BioBank (BCCHB) surveyed Grade 8‐12 students in British Columbia (BC) to determine perceptions before and after the pandemic about topics such as research, pediatric biobanking, vaccinations, and public health policies.Methods: A voluntary, anonymous, 15‐question online survey was distributed through student emails, online school portals, and parent newsletters. Phase 1 of the survey was conducted from May to June 2021 and Phase 2 was conducted from Sept 2021 to June 2022. Questions were edited between Phases 1 and 2 to reflect vaccine availability for adolescents, and the emergence of new variants of COVID‐19.Results: We received a total of 1,022 completed survey responses, from participants aged 12‐19 years in 13 BC school districts and compared the findings to a previous completed school survey [1]. The majority of participants (94%) agreed that COVID‐19 is a serious disease and 95% stated that the pandemic has shown them how important medical research is. While many (95%) agreed on the importance of research before the COVID‐19 pandemic, this increased to 97% after pandemic onset. This desire to contribute to research was reflected by the 80% of participants willing to donate an extra blood sample for biobanking purposes during the COVID‐19 pandemic, compared to 64% from a 2016 school survey conducted among the same age group [1]. Participants (97%) also agreed that research participation would allow them to help others. Of the adolescents surveyed, the majority (85%) were comfortable with making the decision on their own to donate a sample specifically to research. Considering the current public health climate, 92% of participants stated that they will be vaccinated, for reasons such as knowing vaccines are safe, allowing them to engage in social activities, and protecting them from COVID‐19.Conclusion: Adolescents across BC were found to be more willing to participate in research after living through the COVID‐19 pandemic. They also were found to be more willing to donate leftover samples towards biobanking after knowing the importance of research in disease diagnosis and treatment.Reference: [1] Kong CC, Tarling TE, Strahlendorf C, Dittrick M, Vercauteren SM. Opinions of Adolescents and Parents About Pediatric Biobanking. J Adolesc Health, 2016; 58(4):474‐480. doi: 10.1016/j.jadohealth.2015.12.015. PMID: 27013273.O‐11 Tissue Microarrays: Turning Hidden Resources in Research GemsA. Yuksel1, M. Krivanek2, D. R. Catchpoole11Children's Cancer Research Unit, Kids Research, The Children's Hospital at Westmead, SCHN, Westmead, New South Wales, Australia, 2Anatomical Pathology, The Children's Hospital at Westmead, Westmead, New South Wales, AustraliaIt is expected that biobanks provide critical tissue resources to researchers to conduct fundamental investigation into disease states and to ensure the best research use of all tissue biospecimens by linking them to key questions being asked in research. Recognizing the rarity of childhood cancers, small tumour specimen volumes, and the burgeoning need for tissue‐directed research, we describe here the impact of a research‐focussed biospecimen resource. The Tumour Bank at The Children's Hospital at Westmead (TB‐CHW) has initiated alongside the main source of all such tissue, our histopathology department. In 2012 we leveraged the hospital's formalin‐fixed paraffin‐embedded (FFPE) tissue block archive to commence a tissue microarray (TMA) construction program. Our purpose was to provide rationalised access to FFPE tissue whilst not impacting on the availability of blocks for future diagnostic or medico‐legal review. Construction of the TMAs that represented a single childhood cancer subtypes required a deep dive into the block archives with blocks selected covering sample collected over a couple of decade‐long periods or more. This resulted in a tissue resource where enough rare paediatric tumours representing all patients seen at a single centre are dr

Transcription factor ELK1 regulates the expression of histone 3 lysine 9 to affect developmental potential of porcine preimplantation embryos

A series of changes occur in the early embryo that are critical for subsequent development, and the pig is an excellent animal model of human disease, so understanding the regulatory mechanisms of early embryonic development in the pig is of very importance. To find key transcription factors regulating pig early embryonic development, we first profiled the transcriptome of pig early embryos, and confirmed that zygotic gene activation (ZGA) in porcine embryos starts from 4 cell stage. Subsequent enrichment analysis of up-regulated gene motifs during ZGA revealed that the transcription factor ELK1 ranked first. The expression pattern of ELK1 in porcine early embryos was analyzed by immunofluorescence staining and qPCR, and the results showed that the transcript level of ELK1 reached the highest at the 8 cell stage, while the protein level reached the highest at 4 cell stage. To further investigate the effect of ELK1 on early embryo development in pigs, we silenced ELK1 in zygotes and showed that ELK1 silencing significantly reduced cleavage rate, blastocyst rate as well as blastocyst quality. A significant decrease in the expression of the pluripotency gene Oct4 was also observed in blastocysts from the ELK1 silenced group by immunofluorescence staining. Silencing of ELK1 also resulted in decreased H3K9Ac modification and increased H3K9me3 modification at 4 cell stage. To investigate the effect of ELK1 on ZGA, we analyzed transcriptome changes in 4 cell embryos after ELK1 silencing by RNA seq, which revealed that ELK1 silencing resulted in significant differences in the expression of a total of 1953 genes at the 4 cell stage compared with their normal counterparts, including 1106 genes that were significantly upregulated and 847 genes that were significantly downregulated. Through GO and KEGG enrichment, we found that the functions and pathways of down-regulated genes were concentrated in protein synthesis, processing, cell cycle regulation, etc., while the functions of up-regulated genes were focused on aerobic respiration process. In conclusion, this study demonstrates that the transcription factor ELK1 plays an important role in regulation of preimplantation embryo development of pigs and deficiency of ELK1 leads to abnormal epigenetic reprogramming as well as zygotic genome activation, thus adversely affecting embryonic development. This study will provide important reference for the regulation of transcription factors in porcine embryo development.

A fully automated micro‑CT deep learning approach for precision preclinical investigation of lung fibrosis progression and response to therapy

Micro-computed tomography (µCT)-based imaging plays a key role in monitoring disease progression and response to candidate drugs in various animal models of human disease, but manual image processing is still highly time-consuming and prone to operator bias. Focusing on an established mouse model of bleomycin (BLM)-induced lung fibrosis we document, here, the ability of a fully automated deep-learning (DL)-based model to improve and speed-up lung segmentation and the precise measurement of morphological and functional biomarkers in both the whole lung and in individual lobes. µCT-DL whose results were overall highly consistent with those of more conventional, especially histological, analyses, allowed to cut down by approximately 45-fold the time required to analyze the entire dataset and to longitudinally follow fibrosis evolution and response to the human-use-approved drug Nintedanib, using both inspiratory and expiratory μCT. Particularly significant advantages of this µCT-DL approach, are: (i) its reduced experimental variability, due to the fact that each animal acts as its own control and the measured, operator bias-free biomarkers can be quantitatively compared across experiments; (ii) its ability to monitor longitudinally the spatial distribution of fibrotic lesions, thus eliminating potential confounding effects associated with the more severe fibrosis observed in the apical region of the left lung and the compensatory effects taking place in the right lung; (iii) the animal sparing afforded by its non-invasive nature and high reliability; and (iv) the fact that it can be integrated into different drug discovery pipelines with a substantial increase in both the speed and robustness of the evaluation of new candidate drugs. The µCT-DL approach thus lends itself as a powerful new tool for the precision preclinical monitoring of BLM-induced lung fibrosis and other disease models as well. Its ease of operation and use of standard imaging instrumentation make it easily transferable to other laboratories and to other experimental settings, including clinical diagnostic applications.

Methods to Identify Cognitive Alterations from Animals to Humans: A Translational Approach.

The increasing prevalence of cognitive dysfunction and dementia in developed countries, associated with population aging, has generated great interest in characterizing and quantifying cognitive deficits in these patients. An essential tool for accurate diagnosis is cognitive assessment, a lengthy process that depends on the cognitive domains analyzed. Cognitive tests, functional capacity scales, and advanced neuroimaging studies explore the different mental functions in clinical practice. On the other hand, animal models of human diseases with cognitive impairment are essential for understanding disease pathophysiology. The study of cognitive function using animal models encompasses multiple dimensions, and deciding which ones to investigate is necessary to select the most appropriate and specific tests. Therefore, this review studies the main cognitive tests for assessing cognitive deficits in patients with neurodegenerative diseases. Cognitive tests, the most commonly used functional capacity scales, and those resulting from previous evidence are considered. In addition, the leading behavioral tests that assess cognitive functions in animal models of disorders with cognitive impairment are highlighted.

Feline myocardial transcriptome in health and in hypertrophic cardiomyopathy-A translational animal model for human disease.

Hypertrophic cardiomyopathy (HCM) is the most common heart disease in cats, characterized by primary left ventricular hypertrophy. Feline HCM closely resembles human HCM and is suggested as translational animal model for the human disease. A genetic cause is established in humans and suspected for cats, but little is known about the gene expression and pathways involved in the pathogenesis of HCM. To investigate the myocardial transcriptome changes in HCM, RNA sequencing was conducted on left ventricle (LV) and left atrium (LA) samples of healthy cats and cats with HCM (each n = 5; 20 samples). Ingenuity Pathway Analysis was used to determine functional pathways, regulators, and networks. Distinct gene expression profiles were identified in the LV and LA of the feline healthy and HCM myocardium. Analysis of differentially expressed mRNAs (>2 fold; FDR < 0.01) found chamber-specific (LV vs. LA) expression in both healthy and HCM groups, with higher transcriptional activity in the LA. Genes that contribute to the distinct structure and function of each chamber in health and HCM were identified in the regional comparison. The gene expression profiles of HCM compared to healthy hearts revealed disease related genes, including THBS4 and KLHL33 (LV), FAM177B and THRSP (LA), the latter 3 have not been reported for the myocardium so far, as the top differently expressed genes in the HCM heart. Differently expressed genes and functional pathways found in the HCM heart are associated with cardiac remodeling and fibrosis, inflammation, microvascular changes, calcium signaling and cardiac metabolism, with some regional differences. RhoGDI-RhoGTPase signaling, integrin and ILK signaling pathways, the LXR/RXR pathway in the LA, and the PPARα/RXRα, HIF1α and CXCR4 pathways in the LV might be of particular importance in the HCM disease process. This study identified region-specific myocardial gene transcription patterns as well as novel genes and pathways associated with HCM.

Effect of Parental Age, Parity, and Pairing Approach on Reproduction in Strain 13/N Guinea Pigs (Cavia porcellus)

Guinea pigs are important animal models for human disease, and both outbred and inbred lines are utilized in biomedical research. The optimal maintenance of guinea pig colonies, commercially and in research settings, relies on robust informed breeding programs, however, breeding data on specialized inbred strains are limited. Here, we investigated the effects of parental age, parity, and pairing approaches on mean total fetus count, percentage of female pups in the litter, and pup survival rate after 10 days in strain 13/N guinea pigs. Our analysis of colony breeding data indicates that the average litter size is 3.3 pups, with a 25.2% stillbirth rate, a failure-to-thrive outcome in 5.1% of pups, and a 10 day survival rate of 69.7%. The only variable to significantly affect the reproductive outcomes examined was parental age (p < 0.05). In comparison to adults, both juvenile and geriatric sows had lower total fetus counts; juvenile boars had a higher percentage of females in litters, and geriatric boars had a lower 10 day survival rate of pups. These studies provide valuable information regarding the reproductive characteristics of strain 13/N guinea pigs, and support a variety of breeding approaches without significant effects on breeding success.