Disease Models Research Articles

Isoflurane Titration Improves Detection of Hippocampal Lactate by 1H-MRS

Abstract Lactate has increasingly been recognized as both an important fuel source and signaling molecule within the brain. Alterations in brain lactate levels are associated with various neurological diseases. Thus, there is great interest in the in vivo detection and measurement of cerebral lactate levels in animals used for investigation of normal brain function and models of disease. Proton magnetic resonance spectroscopy (1H-MRS) is a non-invasive technique used to measure lactate and other metabolites within the brain. However, lactate can be difficult to detect with conventional 1H-MRS due to its low abundance and spectral overlap with lipids. In addition, volatile anesthetics used during image acquisition increase lactate production, potentially masking any subtle physiological changes in lactate levels. Here we made use of a transgenic mouse model in which expression of lactate dehydrogenase A (Ldha), the rate-limiting enzyme of lactate production, was induced within cortical and hippocampal neurons. Unexpectedly, 1H-MRS analysis under typical isoflurane-induced anesthesia of 4% induction followed by 1.6-2% maintenance, revealed no significant elevation of hippocampal lactate levels in neuronal Ldha induction mice compared to control mice. In contrast, 1H-MRS analysis using an isoflurane titration protocol in which mice were sequentially exposed to 1.6%, 2%, then finally 3% isoflurane, revealed significantly higher hippocampal lactate levels in Ldha transgenic mice compared to controls. In addition, significantly fewer mice were required to detect differences in lactate levels using the isoflurane titration protocol compared to conventional isoflurane-induced anesthesia. Our findings highlight the importance of controlling for the effects of anesthesia when detecting changes in hippocampal lactate levels in vivo and offer a novel protocol for enhanced cerebral lactate detection.

Proteomic analysis of the SMN complex reveals conserved and etiologic connections to the proteostasis network

IntroductionMolecular chaperones and co-chaperones are highly conserved cellular components that perform a variety of duties related to the proper three-dimensional folding of the proteome. The web of factors that carries out this essential task is called the proteostasis network (PN). Ribonucleoproteins (RNPs) represent an underexplored area in terms of the connections they make with the PN. The Survival Motor Neuron (SMN) complex is an assembly chaperone and serves as a paradigm for studying how specific RNAs are identified and paired with their client substrate proteins to form RNPs. SMN is the eponymous component of a large complex, required for the biogenesis of uridine-rich small nuclear ribonucleoproteins (U-snRNPs), that localizes to distinct membraneless organelles in both the nucleus and cytoplasm of animal cells. SMN protein forms the oligomeric core of this complex, and missense mutations in the human SMN1 gene are known to cause Spinal Muscular Atrophy (SMA). The basic framework for understanding how snRNAs are assembled into U-snRNPs is known. However, the pathways and mechanisms used by cells to regulate their biogenesis are poorly understood.MethodsGiven the importance of these processes to normal development as well as neurodegenerative disease, we set out to identify and characterize novel SMN binding partners. We carried out affinity purification mass spectrometry (AP-MS) of Drosophila SMN complexes using fly lines exclusively expressing either wildtype or SMA-causing missense alleles.ResultsBioinformatic analyses of the pulldown data, along with comparisons to proximity labeling studies carried out in human cells, revealed conserved connections to at least two other major chaperone systems including heat shock folding chaperones (HSPs) and histone/nucleosome assembly chaperones. Notably, we found that heat shock cognate protein Hsc70-4 and other HspA family members preferentially associated with SMA-causing alleles of SMN.DiscussionHsc70-4 is particularly interesting because its mRNA is aberrantly sequestered by a mutant form of TDP-43 in mouse and Drosophila ALS (Amyotrophic Lateral Sclerosis) disease models. Most important, a missense allele of Hsc70-4 (HspA8 in mammals) was recently identified as a bypass suppressor of the SMA phenotype in mice. Collectively, these findings suggest that chaperone-related dysfunction lies at the etiological root of both ALS and SMA.

Personalized Nutrition for Obesity Management: A Mini-Review

Abstract Obesity is a pro-inflammatory state and a chronic health condition, which falls within the spectrum of immune-mediated inflammatory diseases (IMIDs). It is caused mainly by the consumption of excess calories and egregious food substances. A Personalized Food Avoidance Dietary Approach for Management of IMIDs (PFA-DAMI) addresses primary and secondary immune intolerances of different individuals to dietary constituents. Exposure to toxic food substances is responsible for inflammatory immune dysfunction that mediates obesity. Hence, a PFA-DAMI is promising for the management of obesity. T cell dysfunction disease mediating models describing inflammatory disease processes underlying IMIDs exist. However, they do not highlight inflammatory processes underlying the diseases concerning toxin-mediated epigenetic T cell dysfunction. Online searches were conducted on databases, such as Google Scholar, PubMed, Biomed Central, and SciELO. Articles were reviewed using keywords, such as obesity, personalized nutrition, immune optimization/dysfunction, T lymphocyte activation/dysfunction, cytokines, and adipokines. There is a putative T cell toxin-mediated dysfunction disease model for IMIDs, which may apply to other diseases. The putative disease model may highlight the actual inflammatory immune dysfunctional processes underlying T cell disease mediation in inflammatory diseases, which may be validated by multiomic studies. Validation of the putative disease model using obesity as an example should pave the way for a better understanding of the role of personalized nutrition in obesity management.

Analysis of Brain Protein Stability Changes in a Mouse Model of Alzheimer's Disease.

The stability of proteins from rates of oxidation (SPROX), thermal proteome profiling (TPP), and limited proteolysis (LiP) techniques were used to profile the stability of ∼2500 proteins in hippocampus tissue cell lysates from 2- and 8-months-old wild-type (C57BL/6J; n = 7) and transgenic (5XFAD; n = 7) mice with five Alzheimer's disease (AD)-linked mutations. Approximately 200-500 protein hits with AD-related stability changes were detected by each technique at each age point. The hit overlap from technique to technique was low, and all of the techniques generated protein hits that were more numerous and largely different from those identified in protein expression level analyses, which were also performed here. The hit proteins identified by each technique were enriched in a number of the same pathways and biological processes, many with known connections to AD. The protein stability hits included 25 high-value conformation biomarkers with AD-related stability changes detected using at least 2 techniques at both age points. Also discovered were subunit- and age-specific AD-related stability changes in the proteasome, which had reduced function at both age points. The different folding stability profiles of the proteasome at the two age points are consistent with a different mechanism for proteasome dysfunction at the early and late stages of AD.

Survival benefit of liver transplantation utilizing marginal donor organ according to ABO blood type.

The current liver transplantation (LT) allocation policy focuses on the Model for End-Stage Liver Disease (MELD) scores, often overlooking factors like blood type and survival benefits. Understanding blood types' impact on survival benefits is crucial for optimizing the MELD 3.0 classification. This study used the United Network for Organ Sharing national registry database (2003-2020) to identify LT characteristics per ABO blood type and to determine the optimal MELD 3.0 scores for each blood type, based on survival benefits. The study included LT candidates aged 18 years or older listed for LT (total N=150,815; A:56,546, AB:5,841, B:18,500, O:69,928). Among these, 87,409 individuals (58.0%) underwent LT (A:32,156, AB:4,362, B:11,786, O:39,105). Higher transplantation rates were observed in AB and B groups, with lower median MELD 3.0 scores at transplantation (AB:21, B:24 vs. A/O:26, p<0.01) and shorter waiting times (AB:101 days, B:172 days vs. A:211 days, O:201 days, p<0.01). A preference for Donation after Cardiac Death (DCD) was seen in A and O recipients. Survival benefit analysis indicated that B blood type required higher MELD 3.0 scores for transplantation than A and O (Donation after Brain Death transplantation: ≥15 in B vs. ≥11 in A/O; DCD transplantation: ≥21 in B vs. ≥11 in A, ≥15 in O). The study suggests revising the allocation policy to consider blood type for improved post-LT survival. This calls for personalized LT policies, recommending higher MELD 3.0 thresholds, particularly for individuals with type B blood.

Synergistic association of Aβ and tau pathology with cortical neurophysiology and cognitive decline in asymptomatic older adults.

Animal and computational models of Alzheimer's disease (AD) indicate that early amyloid-β (Aβ) deposits drive neurons into a hyperactive regime, and that subsequent tau depositions manifest an opposite, suppressive effect as behavioral deficits emerge. Here we report analogous changes in macroscopic oscillatory neurophysiology in the human brain. We used positron emission tomography and task-free magnetoencephalography to test the effects of Aβ and tau deposition on cortical neurophysiology in 104 cognitively unimpaired older adults with a family history of sporadic AD. In these asymptomatic individuals, we found that Aβ depositions colocalize with accelerated neurophysiological activity. In those also presenting medial-temporal tau pathology, linear mixed effects of Aβ and tau depositions indicate a shift toward slower neurophysiological activity, which was also linked to cognitive decline. We conclude that early Aβ and tau depositions relate synergistically to human cortical neurophysiology and subsequent cognitive decline. Our findings provide insight into the multifaceted neurophysiological mechanisms engaged in the preclinical phases of AD.

The p53 target DRAM1 modulates calcium homeostasis and ER stress by promoting contact between lysosomes and the ER through STIM1

It is well established that DNA Damage Regulated Autophagy Modulator 1 (DRAM1), a lysosomal protein and a target of p53, participates in autophagy. The cellular functions of DRAM1 beyond autophagy remain elusive. Here, we show p53-dependent upregulation of DRAM1 in mitochondrial damage-induced Parkinson's disease (PD) models and exacerbation of disease phenotypes by DRAM1. We find that the lysosomal location of DRAM1 relies on its intact structure including the cytosol-facing C-terminal domain. Excess DRAM1 disrupts endoplasmic reticulum (ER) structure, triggers ER stress, and induces protective ER-phagy. Mechanistically, DRAM1 interacts with stromal interacting molecule 1 (STIM1) to tether lysosomes to the ER and perturb STIM1 function in maintaining intracellular calcium homeostasis. STIM1 overexpression promotes cellular health by restoring calcium homeostasis, ER stress response, ER-phagy, and AMP-activated protein kinase (AMPK)-Unc-51 like autophagy activating kinase 1 (ULK1) signaling in cells with excess DRAM1. Thus, by promoting organelle contact between lysosomes and the ER, DRAM1 modulates ER structure and function and cell survival under stress. Our results suggest that DRAM1 as a lysosomal protein performs diverse roles in cellular homeostasis and stress response. These findings may have significant implications for our understanding of the role of the p53/DRAM1 axis in human diseases, from cancer to neurodegenerative diseases.

PROBLEMS OF MANAGING THE IMMUNE RESPONSE UNDER CONDITIONS OF COMPETITIVE ADSORPTION, DIFFUSION PERTURBATIONS AND TEMPERATURE RESPONSE OF THE ORGANISM

Analysis of the problems associated with the rapid spread of infectious diseases indicates the need for new approaches to the diagnosis and develop- ment of effective personalized treatment programs. One of the important aspects of solving this problem is the creation of a mathematical modeling toolkit for predicting the dynamics of infectious diseases, taking into account spatial effects, concentrated effects, various mechanisms of body protec- tion in the conditions of the application of various types of therapeutic methods of treatment. An important component of complex methods of treat- ment of a wide range of infectious diseases is the use of adsorption drugs, which, along with detoxification of the body, are able to provide an addi- tional mechanism of neutralization of viral elements. In the process of using such means, not only pathogenic elements and toxins will be adsorbed, but also various types of immune agents, which will affect the dynamics of the immune response. In the paper the infectious disease model is generalized to take into account the features of competitive adsorption of antigens and immune agents in the conditions of diffusion perturbations, concentrated influences and temperature reaction of the body. By synthesizing the ideas of step-by-step procedures, asymptotic and numerical methods, an effective computational technology of step-by-step approximation of the solution of the original model singularly perturbed problem was built. The results of the computer experiments presented in the paper illustrate the features of reducing the predicted concentration of viral elements during their competi- tive adsorption together with immune agents, in particular, the effect of reducing the efficiency of the use of non-specific adsorbents. It is emphasized that taking into account the features of the action of competitive adsorption is important for making decisions regarding the formation of rational treatment programs with the complex use of adsorbing substances and immunological drugs.

Inhibiting Ca2+ channels in Alzheimer's disease model mice relaxes pericytes, improves cerebral blood flow and reduces immune cell stalling and hypoxia.

Early in Alzheimer's disease (AD), pericytes constrict capillaries, increasing their hydraulic resistance and trapping of immune cells and, thus, decreasing cerebral blood flow (CBF). Therapeutic approaches to attenuate pericyte-mediated constriction in AD are lacking. Here, using in vivo two-photon imaging with laser Doppler and speckle flowmetry and magnetic resonance imaging, we show that Ca2+ entry via L-type voltage-gated calcium channels (CaVs) controls the contractile tone of pericytes. In AD model mice, we identifed pericytes throughout the capillary bed as key drivers of an immune reactive oxygen species (ROS)-evoked and pericyte intracellular calcium concentration ([Ca2+]i)-mediated decrease in microvascular flow. Blocking CaVs with nimodipine early in disease progression improved CBF, reduced leukocyte stalling at pericyte somata and attenuated brain hypoxia. Amyloid β (Aβ)-evoked pericyte contraction in human cortical tissue was also greatly reduced by CaV block. Lowering pericyte [Ca2+]i early in AD may, thus, offer a therapeutic strategy to enhance brain energy supply and possibly cognitive function in AD.

Validation, Diagnostic, and Monitoring Roles of Multiomic Studies in Putative Immunotoxiepigenetic Models of Immune-Mediated Inflammatory Diseases

Abstract Immune-mediated inflammatory diseases (IMIDs) are mediated by immune dysfunction. Putative models of T-cell-mediated immune dysfunction in IMIDs exist and are yet to be validated by multiomics studies. Prompt diagnosis and monitoring are crucial for IMID management. The aim of this review was to discuss the putative T-cell dysfunction model of IMIDs and the role of multiomics studies for validation of the disease models as well as its diagnostic and monitoring potential in IMIDs. Online searches on databases such as Google Scholar, PubMed, Biomed Central, and SciELO were carried out. An attempt was made to review articles with keywords such as genomics, proteomics, transcriptomics, metabolomics, T cell immunopathogenesis/dysfunction, immune dysfunction, and IMIDs. The putative T-cell dysfunction model of IMIDs and its validation with multiomics studies were discussed. The diagnostic and monitoring roles of multiomic studies for management of IMIDs were also discussed. The review concluded that multiomic studies have validating potential for the putative T-cell dysfunction model of IMIDs as well as diagnostic and monitoring roles in IMIDs.

Tet1-mediated activation of the Ampk signaling by Trpv1 DNA hydroxymethylation exerts neuroprotective effects in a rat model of Parkinson's disease.

Epigenetic regulation plays a role in Parkinson's disease (PD), and ten-eleven translocation methylcytosine dioxygenase 1 (TET1) catalyzes the first step in DNA demethylation by converting 5-methylcytosine to 5-hydroxymethylcytosine. We investigated whether TET1 binds to the promoter of the transient receptor potential cation channel subfamily V member 1 (TRPV1) and regulates its expression, thereby controlling oxidative stress in PD. TRPV1 was identified as an oxidative stress-associated gene in the GSE20186 dataset including substantia nigra from 14 patients with PD and 14 healthy controls and the Genecards database. Lentiviral vectors were used to manipulate Trpv1 expression in rats, followed by 6-hydroxydopamine hydrochloride (6-OHDA) injection for modeling. Behavioral tests, immunofluorescence, Nissl staining, western blot assays, DHE fluorescent probe, biochemical analysis, and ELISA were conducted to assess oxidative stress and neurotoxicity. Trpv1 expression was significantly reduced in the brain tissues of 6-OHDA-treated Parkinsonian rats. Trpv1 alleviated behavioral dysfunction, oxidative stress, and dopamine neuron loss in rats. TET1 mediated TRPV1 hydroxymethylation to promote its expression, and Trpv1 inhibition reversed the mitigating effect of Tet1 on oxidative stress and behavioral dysfunction in PD. TRPV1 activated the AMPK signaling by promoting AMPK phosphorylation to alleviate neurotoxicity and oxidative stress in SH-SY5Y cells. Tet1-mediated Trpv1 hydroxymethylation modification promotes the Ampk signaling activation, thereby eliciting neuroprotection in 6-OHDA-treated Parkinsonian rats. These findings provide experimental evidence that targeting the TET1/TRPV1 axis may be neuroprotective for PD by acting on the AMPK signaling.

Analyzing the dynamics and optimal control of a vector-borne model with dual vertical transmission and multiple serotypes

Abstract Transmission and contact patterns have an important influence on the intensity and trend of disease transmission. Taking into account two different serotypes and the vertical transmission of the disease in both humans and mosquitoes, this paper proposes a novel vector-borne disease model. In order to explore the evolution mechanisms of the model and the impacts of contact pattern on disease transmission. The basic regeneration number of the proposed model is obtained via the next-generation matrix approach. Subsequently, we analyze the conditions of the stabilities and existences of the equilibrium points. Further, the sensitivity of the parameters is studied using numerical simulations. It is observe that the increase of vertical transmission rate of mosquitoes leads to the increase of the basic reproduction number. In addition, we develop vaccination and insecticide control strategies and derive the existence and expression of the optimal solution. Our numerical results show that vaccination has a little effect on preventing the spread of disease among mosquitoes. However, insecticide spraying is effective in reducing the spread of disease between humans and mosquitoes.&amp;#xD;

Advances in melt electrowriting for cardiovascular applications

Melt electrowriting (MEW) is an electric-field-assisted additive biofabrication technique that has brought significant advancements to bioinspired scaffold design for soft tissue engineering and beyond. Owing to its targeted microfiber placement, MEW has become a powerful platform technology for the fabrication of in vitro disease models up to functional biohybrid constructs that are investigated in vivo to reach clinical translation soon. This work provides a concise overview of this rapidly evolving field by highlighting the key contributions of MEW to cardiovascular tissue engineering. Specifically, we i) pinpoint the methods to introduce microvascular networks in thick 3D constructs benefitting from (sacrificial) MEW microfibers, ii) report MEW-based concepts for small-diameter vascular grafts and stents, iii) showcase how contracting cardiac tissues can profit from the tunable structure–property relationship of MEW scaffolds, and iv) address how complete regenerative heart valves can be built on complex fiber scaffold architectures that recapitulate J-shaped tensile properties and tissue heterogeneity. Lastly, we touch on novel biomaterial advancements and discuss the technological challenges of MEW to unlock the full potential of this transformative technology.

Maternal Ketone Supplementation Throughout Gestation Improves Neonatal Cardiac Dysfunction Caused by Perinatal Iron Deficiency.

Iron deficiency (ID) is common during gestation and in early infancy and has been shown to adversely affect cardiac development and function, which could lead to lasting cardiovascular consequences. Ketone supplementation has been shown to confer cardioprotective effects in numerous disease models. Here we tested the hypothesis that maternal ketone supplementation during gestation would mitigate cardiac dysfunction in ID neonates. Female Sprague Dawley rats were fed an iron-restricted or iron-replete diet before and throughout pregnancy. Throughout gestation, iron-restricted dams were given either a daily subcutaneous injection of ketone solution (containing β-hydroxybutyrate [βOHB]) or saline (vehicle). Neonatal offspring cardiac function was assessed by echocardiography at postnatal days (PD)3 and 13. Hearts and livers were collected post-mortem for assessments of mitochondrial function and gene expression profiles of markers oxidative stress and inflammation. Maternal iron restriction caused neonatal anemia and asymmetric growth restriction at all time points assessed, and maternal βOHB treatment had no effect on these outcomes. Echocardiography revealed reduced ejection fraction despite enlarged hearts (relative to body weight) in ID offspring, resulting in impaired oxygen delivery, which was attenuated by maternal βOHB supplementation. Further, maternal ketone supplementation affected biochemical markers of mitochondrial function, oxidative stress and inflammation in hearts of neonates, implicating these pathways in the protective effects conferred by βOHB. In summary, βOHB supplementation confers protection against cardiac dysfunction in ID neonates, and could have implications for the treatment of anemic babies.

Palmatine ameliorates N-methyl-N'-nitrosoguanidine-induced chronic atrophic gastritis through the STAT1/CXCL10 axis.

Chronic atrophic gastritis (CAG) is a prevalent preneoplastic condition of the stomach. Palmatine (PAL), an isoquinoline alkaloid isolated from Rhizoma Coptidis (RC), has significant anti-inflammatory properties and is often used to treat gastrointestinal disorders. However, the mechanism of PAL on CAG remains unclear. In this study, N-methyl-N'-nitrosoguanidine (MNNG) was used to induce CAG inflammatory disease models invivo and invitro. The efficacy of five alkaloids in RC and the dose-dependent effects of the most effective PAL in CAG mice were evaluated in two animal experiments. RNA-seq and western blot revealed that PAL significantly improved IL-17, TNF, and NF-kappa B inflammation-related signaling pathways. Further hub gene prediction and experimental validation revealed that PAL modulated the STAT1/CXCL10 axis, thereby exerting attenuation of CAG through the regulation of IL-17, TNF-α, and p-p65 expression. In conclusion, PAL was proposed to mitigate MNNG-induced CAG, potentially through the inhibition of oxidative stress and inflammatory responses via the STAT1/CXCL10 axis. This approach is an effective complement to the use of PAL in the treatment of CAG.

Melatonin Ameliorates Abnormal Sleep-Wake Behavior via Facilitating Lipid Metabolism in a Zebrafish Model of Parkinson's Disease.

Sleep-wake disorder is one of the most common nonmotor symptoms of Parkinson's disease (PD). Melatonin has the potential to improve sleep-wake disorder, but its mechanism of action is still unclear. Our data showed that melatonin only improved the motor and sleep-wake behavior of a zebrafish PD model when melatonin receptor 1 was present. Thus, we explored the underlying mechanisms by applying a rotenone model. After the PD zebrafish model was induced by 10 nmol/L rotenone, the motor and sleep-wake behavior were assessed. In situ hybridization and real-time quantitative PCR were used to detect the expression of melatonin receptors and lipid-metabolism-related genes. In the PD model, we found abnormal lipid metabolism, which was reversed by melatonin. This may be one of the main pathways for improving PD sleep-wake disorder.

Therapeutic Potential of Crocin and Nobiletin in a Mouse Model of Dry Eye Disease: Modulation of the Inflammatory Response and Protection of the Ocular Surface

Background: Dry eye disease (DED) is a multifactorial condition characterized by ocular surface inflammation, tear film instability, and corneal epithelial damage. Current treatments often provide temporary relief without addressing the underlying inflammatory mechanisms. Objectives: This study examined the therapeutic potential of crocin and nobiletin, two naturally derived compounds with well-known antioxidant and anti-inflammatory properties, in a mouse model of DED induced by lacrimal gland excision (ELG). Methods: Thirty female Balb/c mice were divided into five groups (n = 6 each): Control (sham surgery), untreated DED, nobiletin-treated DED (32.75 µM), crocin-treated DED (34 µM), and 1% betamethasone-treated DED. Treatments were administered three times daily for 28 days. Ocular tissues were evaluated using Hematoxylin and Eosin (H&amp;E) staining and fluorescein staining. Conjunctival inflammatory cytokines, including interleukin-6 (IL-6), interleukin-1 beta (IL-1β), and tumor necrosis factor-alpha (TNF-α), were measured by enzyme-linked immunosorbent assay (ELISA). Results: Histological analysis showed that the crocin and nobiletin treatment groups exhibited reduced epithelial disruption, keratinization, and inflammatory cell infiltration compared to the untreated DED group. The ELISA assay revealed that both compounds efficiently inhibited the production of the pro-inflammatory cytokines IL-6, TNF-α, and IL-1β, which are key mediators of DED pathogenesis. Fluorescein staining further confirmed the protective impact of crocin and nobiletin on corneal epithelial integrity. Moreover, the anti-inflammatory and epithelial-preserving effects of these compounds were comparable to those of the corticosteroid betamethasone. Conclusions: Overall, these findings suggest that crocin and nobiletin have therapeutic potential for DED management by modulating inflammatory responses and enhancing ocular surface healing. These naturally derived compounds offer promising avenues for the development of safer and more effective treatments for this challenging condition. However, further investigations, including clinical trials, are essential to elucidate the underlying mechanisms of action and optimize therapeutic approaches.

Progress in the Study of Animal Models of Metabolic Dysfunction-Associated Steatotic Liver Disease

Metabolic dysfunction-associated steatotic liver disease (MASLD) has recently been proposed as an alternative term to NAFLD. MASLD is a globally recognized chronic liver disease that poses significant health concerns and is frequently associated with obesity, insulin resistance, and hyperlipidemia. To better understand its pathogenesis and to develop effective treatments, it is essential to establish suitable animal models. Therefore, attempts have been made to establish modelling approaches that are highly similar to human diet, physiology, and pathology to better replicate disease progression. Here, we reviewed the pathogenesis of MASLD disease and summarised the used animal models of MASLD in the last 7 years through the PubMed database. In addition, we have summarised the commonly used animal models of MASLD and describe the advantages and disadvantages of various models of MASLD induction, including genetic models, diet, and chemically induced models, to provide directions for research on the pathogenesis and treatment of MASLD.

Sex-specific molecular signature of mouse podocytes in homeostasis and in response to pharmacological challenge with rapamycin

BackgroundSex differences exist in the prevalence and progression of major glomerular diseases. Podocytes are the essential cell-type in the kidney which maintain the physiological blood-urine barrier, and pathological changes in podocyte homeostasis are critical accelerators of impairment of kidney function. However, sex-specific molecular signatures of podocytes under physiological and stress conditions remain unknown. This work aimed at identifying sexual dimorphic molecular signatures of podocytes under physiological condition and pharmacologically challenged homeostasis with mechanistic target of rapamycin (mTOR) inhibition. mTOR is a crucial regulator involved in a variety of physiological and pathological stress responses in the kidney and inhibition of this pathway may therefore serve as a general stress challenger to get fundamental insights into sex differences in podocytes.MethodsThe genomic ROSAmT/mG-NPHS2 Cre mouse model was used which allows obtaining highly pure podocyte fractions for cell-specific molecular analyses, and vehicle or pharmacologic treatment with the mTOR inhibitor rapamycin was performed for 3 weeks. Subsequently, deep RNA sequencing and proteomics were performed of the isolated podocytes to identify intrinsic sex differences. Studies were supplemented with metabolomics from kidney cortex tissues.ResultsAlthough kidney function and morphology remained normal in all experimental groups, RNA sequencing, proteomics and metabolomics revealed strong intrinsic sex differences in the expression levels of mitochondrial, translation and structural transcripts, protein abundances and regulation of metabolic pathways. Interestingly, rapamycin abolished prominent sex-specific clustering of podocyte gene expression and induced major changes only in male transcriptome. Several sex-biased transcription factors could be identified as possible upstream regulators of these sexually dimorphic responses. Concordant to transcriptomics, metabolomic changes were more prominent in males. Remarkably, high number of previously reported kidney disease genes showed intrinsic sexual dimorphism and/or different response patterns towards mTOR inhibition.ConclusionsOur results highlight remarkable intrinsic sex-differences and sex-specific response patterns towards pharmacological challenged podocyte homeostasis which might fundamentally contribute to sex differences in kidney disease susceptibilities and progression. This work provides rationale and an in-depth database for novel targets to be tested in specific kidney disease models to advance with sex-specific treatment strategies.

Synthetic Human Embryos, Embryo Models and Embryo-like Structures in Islam

ABSTRACT A major breakthrough in developmental biology is the ex vivo generation of synthetic human embryos from stem cells. A comprehensive, in-depth bioethical analysis from a Sunni Islamic perspective reveals that the reproductive applications of synthetic human embryos contravene Islamic precepts of preserving lineage integrity (Hifz al-Nasl) due to disruption and confusion of kinship and familial relationships, similar to human cloning with somatic cell nuclear transfer. However, their non-reproductive applications in generating replacement tissues/organs, serving as in vitro experimental models of human development and disease, and testing platforms for evaluating pharmaceuticals and biomedical devices appear to align with Islamic principles.