Organ Level Research Articles

A novel approach to assessing cardiotoxic effects of nanoparticles in a toxicological experiment

Introduction. The study of nanoparticles for potential cardiotoxic effects is a comprehensive multi-stage process based on an integrated approach. Along with generally accepted research methods, molecular biology techniques using modern highly sensitive equipment are being actively introduced into toxicology testing. The aim of the study was to describe a novel approach to assessing cardiotoxic effects of nanoparticles, from the molecular level to the functional response of the whole organism. Materials and methods. Our new approach to assessing cardiotoxic effects of nanoparticles in rats included the examination of changes at the molecular (e.g., the ratio of myosin heavy chains), subcellular (by electron microscopy), cellular and tissue (by histological testing), system and organ (by non-invasive recording of electrocardiogram and blood pressure parameters and biochemical testing of blood serum) levels. We have tested the proposed approach by evaluating lead (PBO) and cadmium oxide (CdO) nanoparticles in rats. Results. Hypotension observed after PbO and/or CdO nanoparticle exposure indicates to the damage to the vascular bed due to penetration and accumulation of the nanoparticles in vascular cells, as well as direct damage to the endothelium, increased oxidative stress, and inflammation. In accordance with the system for assessing nanoparticle-induced cardiotoxicity developed on the basis of toxicology test results, lead and cadmium oxides, both separately and combined, have a pronounced cardiotoxic effect. Limitations. Our work was limited to examining the main indicators of the cardiotoxic effects of nanoparticles in a toxicological experiment on one animal species (rats). Conclusion. The data analysis revealed varying degrees of manifestation of nanoparticle cardiotoxicity, both at the molecular level and at the intracellular, cellular, tissue, organ, and body levels. The use of this approach will allow a better in-depth assessing effects of nano-sized particles on the heart and blood vessels for identification of risks for cardiovascular disease.

Assessment of disorders of metabolic process on the base of quantification of target proteins under aerogenic influence of aluminum oxide in children

Introduction. The study of the mechanisms of the occurrence of adverse events at the molecular level, followed by the study of biological processes at the cellular, tissue, and organ level, allows further investigating the mechanism of the toxic action of chemicals to predict the development of adverse effects in humans. The purpose of the study is to evaluate disturbances in the signal-transporter pathways of metabolic processes on the base of the quantification of identified target proteins under aerogenic exposure to aluminum oxide in children. Materials and methods. Using the methods of chemical-analytical, statistical, proteomic, biochemical, histological research, and bioinformation analysis, the adverse effects were assessed in 4–7 years children and Wistar rats exposed to aerogenic and inhalation exposure to aluminum oxide (Al2O3). A comparative analysis of the results obtained in the experiment and natural conditions was carried out. Results. In children under conditions of long-term aerogenic exposure to Al2O3 at the level of 0.1–1.0 RfC, the urine concentration of the studied substance was found to be up to 2 times higher than the average comparison value and the reference value. In the experiment with chronic inhalation of Al2O3 at a dose equivalent to the real one, the aluminum urine content in rats was 3.5 times higher than the control value. Comparative analysis of proteomic maps in children revealed 23 significantly different protein spots, 8 of which had a relationship between intensity changes and an increase in Al urine concentration. In rats, 15 significantly different protein spots were found between the groups, 13 of which had a reliable relationship with the exposure marker. In natural and experimental studies, only two identical proteins were found: apolipoprotein A-I and transthyretin; increased levels of ALAT, ASAT and alkaline phosphatase, total and direct bilirubin, gamma-aminobutyric and glutamic acids, lipid hydroperoxide in the blood serum; increased MDA and decreased AOA in the blood plasma. Pathomorphological changes in the tissues of the brain, heart, and liver were confirmed in the experiment. Limitations. The research conducted allows drawing a conclusion about the effect of Al2O3 on the body only through the aerogenic route of entry. Conclusion. Based on the bioinformational analysis of the results obtained and assessment of the cause-and-effect relationships of the transformation of the proteomic profile of blood plasma under natural conditions, verified in the experiment, the leading molecular-cellular events in the development of adverse effects in the form of oxidation, imbalance of lipoprotein and neurotransmitter metabolism, and decreased neurogenesis activity were identified. Metabolic disorders are predicted in the tissues of the heart, blood vessels, liver, and brain under continued conditions of aerogenic exposure to Al2O3. The study of the cascade of events of adverse responses (from the molecular to the organ level) expands knowledge about the pathogenetic mechanisms of metabolic processes of dysregulation of signal-transporter pathways in the human body in response to the influence of a chemical factor, including Al2O3. This increases the effectiveness of early prediction of the occurrence of the disease and the development of targeted measures to prevent adverse consequences.

Mechanism of Coronary Microcirculation Obstruction after Acute Myocardial Infarction and Cardioprotective Strategies.

ST-segment elevation myocardial infarction patients are best treated with emergency percutaneous coronary intervention (PCI), while coronary microvascular dysfunction and obstruction (CMVO) are indicated by the absence or slowing of antegrade epicardial flow on angiography, resulting in suboptimal myocardial perfusion despite the lack of mechanical vascular obstruction. CMVO occurs in up to half of patients who undergo PCI for the first time and is associated with poor outcomes. This review summarizes the complex mechanisms leading to CMVO and elaborates on the changes observed at the organism, tissue, organ, cellular, and molecular levels. It also describes the current diagnostic methods and comprehensive treatment methods for CMVO.

NAD+-boosting agent nicotinamide mononucleotide potently improves mitochondria stress response in Alzheimer’s disease via ATF4-dependent mitochondrial UPR

Extensive studies indicate that mitochondria dysfunction is pivotal for Alzheimer’s disease (AD) pathogenesis; while cumulative evidence suggests that increased mitochondrial stress response (MSR) may mitigate neurodegeneration in AD, explorations to develop a MSR-targeted therapeutic strategy against AD are scarce. We combined cell biology, molecular biology, and pharmacological approaches to unravel a novel molecular pathway by which NAD+-boosting agent nicotinamide mononucleotide (NMN) regulates MSR in AD models. Here, we report dyshomeostasis plasma UPRmt-mitophagy-mediated MSR profiles in AD patient samples. NMN restores NAD+ metabolic profiles and improves MSR through the ATF4-dependent UPRmt pathway in AD-related cross-species models. At the organismal level, NAD+ repletion with NMN supplementation ameliorates mitochondrial proteotoxicity, decreases hippocampal synaptic disruption, decreases neuronal loss, and brain atrophy in mice model of AD. Remarkably, omics features of the hippocampus with NMN show that NMN leads to transcriptional changes of genes and proteins involved in MSR characteristics, principally within the astrocyte unit rather than microglia and oligodendrocytes. In brief, our work provides evidence that MSR has an active role in the pathogenesis of AD, as reducing mitochondrial homeostasis via atf4 depletion in AD mice aggravates the hallmarks of the disease; conversely, bolstering mitochondrial proteostasis by NMN decreases protein aggregation, restores memory performance, and delays disease progression, ultimately translating to increased healthspan.

A concentric 3D-printed centrifuge rotor to study the effects of hypergravity on small organisms at three different but concurrent radial accelerations

For life to reach space, it must first escape Earth's gravity—hence exposure to hypergravity during launch, and interplanetary acceleration and deceleration. Long-term colonisation of space requires the establishment of sustainable, in situ, food production systems. Earthworms may be involved as a regolith/soil ameliorant, a source of protein, and for organic waste conversion. Earthworms, however, have hydrostatic skeletons that may be susceptible to hypergravity exposure. We investigated whether earthworms (Eisenia andrei) would survive prolonged centrifugal hypergravity using a table-top centrifuge. Our intention was to concurrently test three different hypergravity treatments (plus a 1 G control). Our concentric rotor concept, with appropriate design adaptations, allows concurrent testing of three different gravity treatments on small- and microorganisms, cell cultures, plants, microcosm, and chemical/biochemical reactions for extended periods of hours to months. Our concentric concept therefore is easily adaptable and may facilitate other terrestrial-based research on chemical, molecular, and organismal levels.•We designed a 3D-printed centrifuge rotor with three concentric rings, each ring with four separate 125 mL compartments.•This design was tested with an 8-day pilot run with four young worms per container at 1.7 G, 2.5 G, and 3.1 G, with a 1 G control.•All worms at all treatments gained body mass, the mean increases of which were almost indistinguishable between treatments, illustrating the utility of the design.

Functionally Modified Graphene Oxide as an Alternative Nanovehicle for Enhanced dsRNA Delivery in Improving RNAi-Based Insect Pest Control.

RNA interference (RNAi) has shown substantial promise as a sustainable pest management solution. However, the efficacy of RNAi-based insecticides heavily relies on advanced nanocarrier-mediated delivery systems. In this study, we modified raw graphene oxide into positively charged nanocarriers (GONs) tailored to bind with double-stranded RNA (dsRNA). The resulting GONs@dsRNA complexes demonstrated a small particle size (106 nm) and maintained stability under various conditions, including insect gut extracts, extreme pH, and extreme temperature. Furthermore, GONs efficiently transported dsRNA molecules into Drosophila S2 cells and Lepidoptera Sf9 cells, leading to an enhanced target transcript knockdown. Targeting the vacuolar ATPase gene, vha26, induced significant mortality and target transcript knockdown in D. suzukii adults but not in S. exigua. Finally, GONs@dsRNA complexes exhibited negligible cytotoxicity at both the cellular and organismal levels. This study demonstrates the potential of GONs as a biosafe nanovehicle for efficient dsRNA delivery into insects, presenting an alternative strategy for advancing RNAi applications in fundamental studies and pest control.

Dissection of Mosquito Ovaries, Midgut, and Salivary Glands for Microbiome Analyses at the Organ Level.

The global burden of mosquito-transmitted diseases, including malaria, dengue, West Nile, Zika, Usutu, and yellow fever, continues to increase, posing a significant public health threat. With the rise of insecticide resistance and the absence of effective vaccines, new strategies are emerging that focus on the mosquito's microbiota. Nevertheless, the majority of symbionts remain resistant to cultivation. Characterizing the diversity and function of bacterial genomes in mosquito specimens, therefore, relies on metagenomics and subsequent assembly and binning strategies. The obtention and analysis of Metagenome-Assembled Genomes (MAGs) from separated organs can notably provide key information about the specific role of mosquito-associated microbes in the ovaries (the reproductive organs), the midgut (key for food digestion and immunity), or the salivary glands (essential for the transmission of vector-borne diseases as pathogens must colonize them to enter the saliva and reach the bloodstream during a blood meal). These newly reconstructed genomes can then pave the way for the development of novel vector biocontrol strategies. To this aim, it is required to isolate mosquito organs while avoiding cross-contamination between them or with microorganisms present in other mosquito organs. Here, we describe an optimized and contamination-free dissection protocol for studying mosquito microbiome at the organ level.

Lipid Droplets Big and Small: Basic Mechanisms That Make Them All.

Lipid droplets (LDs) are dynamic storage organelles with central roles in lipid and energy metabolism. They consist of a core of neutral lipids, such as triacylglycerol, which is surrounded by a monolayer of phospholipids and specialized surface proteins. The surface composition determines many of the LD properties, such as size, subcellular distribution, and interaction with partner organelles. Considering the diverse energetic and metabolic demands of various cell types, it is not surprising that LDs are highly heterogeneous within and between cell types. Despite their diversity, all LDs share a common biogenesis mechanism. However, adipocytes have evolved specific adaptations of these basic mechanisms, enabling the regulation of lipid and energy metabolism at both the cellular and organismal levels. Here, we discuss recent advances in the understanding of both the general mechanisms of LD biogenesis and the adipocyte-specific adaptations controlling these fascinating organelles.

Evolution of Repetitive Elements, Their Roles in Homeostasis and Human Disease, and Potential Therapeutic Applications.

Repeating sequences of DNA, or repetitive elements (REs), are common features across both prokaryotic and eukaryotic genomes. Unlike many of their protein-coding counterparts, the functions of REs in host cells remained largely unknown and have often been overlooked. While there is still more to learn about their functions, REs are now recognized to play significant roles in both beneficial and pathological processes in their hosts at the cellular and organismal levels. Therefore, in this review, we discuss the various types of REs and review what is known about their evolution. In addition, we aim to classify general mechanisms by which REs promote processes that are variously beneficial and harmful to host cells/organisms. Finally, we address the emerging role of REs in cancer, aging, and neurological disorders and provide insights into how RE modulation could provide new therapeutic benefits for these specific conditions.

Effects of Aerobic Exercise on Vascular Wall Thickness: A Meta-Analysis of Pre-Clinical Trials on Rat Model.

Exercise instigates adaptations in structural as well as functional organisation of the body at tissue and organ levels. However, the effect of exercise on vascular wall morphology is still under investigation. The objective of the review is to provide a meta-analysis of data from the previously published research to provide compiled analysis to determine the effect of exercise on the thickness of large-sized vessel walls. This meta-analysis was executed at the Ziauddin College of Rehabilitation Sciences with an extensive literature search using databases and search engines such as Pubmed, CINAHL, Web of Science, Scopus, and Google Scholar between September 2022 and January 2023. Studies were appraised according to an estimate of mean difference and pooled effect size was calculated using a random effect model. The risk of bias was also calculated using SYRCLE's risk of bias tool and CAMARADES. Overall, five eligible pre-clinical trials using rat models with pertinent data to determine the effect of exercise on vessel wall thickness were included in the meta-analysis. The results showed that aerobic exercise significantly reduced the vessel wall thickness (SMD -0.854 5%; CI: -1.365 to -0.344; p <0.001). Moreover, no significant publication bias was found through funnel plot and statistical test (Egger's p = 0.276; Begg's p = 0.624). However, an extensive review of the literature currently available on the topic has shown mixed findings that are consistent with the study and vice versa. Therefore, it indicated the need for sub-group analysis with different types of exercises for more peculiar clinical approaches. Key Words: Morphometry, Animal studies meta-analysis, Media thickness, Physical activity, Vascular morphology.

Myogenic Classical Endocannabinoids, Their Targets and Activity.

This review focuses on the recently discovered specific action of two classical endocannabinoids(ECs), 2-arachidonoylglycerol(2-AG) and arachidonoyl ethanolamide(AEA), in the case of their synthesis and degradation in skeletal muscles; in other words, this review is dedicated to properties and action of the myoendocannabinoid (myoEC) pool. Influence of this pool is considered at three different levels: at the level of skeletal muscles, motor synapses, and also at the level of the whole organism, including central nervous system. Special attention is paid to the still significantly underestimated and intriguing ability of ECs to have positive effect on energy exchange and contractile activity of muscle fibers, as well as on transmitter secretion in motor synapses. Role of muscle contractions in regulation of activity balance between the enzymes catalyzing synthesis and degradation of myoECs and, therefore, in the release of myoECs and exertion of their specific effects is thoroughly considered. Increasingly popular hypotheses about the prominent role of myoECs (AEA and/or 2-AG) in the rise of the overall level of ECs in the blood during muscle exercise and the development of "runner's high" and about the role of myoECs in the correction of a number of psychophysiological conditions (pain syndrome, stress, etc.) are discussed here. Thus, this review presents information about the myoEC pool from a totally new viewpoint, underlining its possible independent and non-trivial regulatory role in the body, in contrast to the traditional and well-known activity of neurogenic ECs.

A role for organ level dynamics in morphogenesis of the C. elegans hermaphrodite distal tip cell.

The morphology of cells in vivo can arise from a variety of mechanisms. In the Caenorhabditis elegans hermaphrodite gonad, the distal tip cell (DTC) elaborates into a complex plexus over a relatively short developmental time period, but the mechanisms underlying this change in cell morphology are not well defined. We correlated the time of DTC elaboration with the L4-to-adult molt, but ruled out a relevant heterochronic pathway as a cue for DTC elaboration. Instead, we found that the timing of gonad elongation and aspects of underlying germline flux influence DTC elaboration. We propose a 'hitch and tow' aspect of organ-level dynamics that contributes to cellular morphogenesis, whereby germline flux drags the flexible DTC cell cortex away from its stationary cell body. More broadly, we speculate that this mechanism may contribute to cell shape changes in other contexts with implications for development and disease.

Neuropeptide signaling network of Caenorhabditis elegans: from structure to behavior.

Neuropeptides are abundant signaling molecules that control neuronal activity and behavior in all animals. Owing in part to its well-defined and compact nervous system, Caenorhabditis elegans has been one of the primary model organisms used to investigate how neuropeptide signaling networks are organized and how these neurochemicals regulate behavior. We here review recent work that has expanded our understanding of the neuropeptidergic signaling network in C. elegans by mapping the evolutionary conservation, the molecular expression, the receptor-ligand interactions, and the system-wide organization of neuropeptide pathways in the C. elegans nervous system. We also describe general insights into neuropeptidergic circuit motifs and the spatiotemporal range of peptidergic transmission that have emerged from in vivo studies on neuropeptide signaling. With efforts ongoing to chart peptide signaling networks in other organisms, the C. elegans neuropeptidergic connectome can serve as a prototype to further understand the organization and the signaling dynamics of these networks at organismal level.

Factors Associated with the Outcomes of Patients with Hospital-Acquired Pneumonia (HAP) at Dr. Moewardi General Hospital, Surakarta

Introduction: Hospital-acquired pneumonia (HAP) is a common infection with a poor prognosis. Previous studies on factors influencing HAP outcomes have yielded inconsistent findings. Therefore, further research is needed to determine risk factors that affect HAP outcomes. This study evaluated the factors associated with HAP outcomes to enable timely interventions to reduce mortality, costs, and length of stay (LOS). Methods: This cross-sectional study was conducted at Dr. Moewardi General Hospital, Surakarta, using medical record data from January to December 2022. The data included age, gender, malnutrition, anemia, level of consciousness, comorbidities, bacterial culture, and multidrug-resistant organisms (MDROs). The analyses were performed using the Chi-squared and Mann-Whitney U tests, followed by multiple logistic and linear regression tests to determine the correlation between risk factors and outcomes (recovery, death, and LOS). Results: This study included 102 patients with HAP, the majority being males (64.7%) aged 60 years old and above. Additionally, most patients did not exhibit malnutrition (87.3%) or anemia (96.1%), had normal levels of consciousness (67.6%) and mild comorbidities (60.8%), were not infected with MDROs (66.7%), and were predominantly infected by Klebsiella pneumoniae (21.6%). The multiple logistic regression test revealed that decreased consciousness was significantly associated with increased mortality (p &lt; 0.001). Meanwhile, age 60 years old and above was significantly associated with a shorter LOS (p = 0.05). Conclusion: The majority of HAP cases occurred in men aged 60 years and above. Furthermore, there was a significant relationship between decreased consciousness and increased mortality, as well as between the age of 60 years and above and a shorter LOS.

RRAGD variants cause cardiac dysfunction in a zebrafish model.

The Ras-related GTP-binding protein D (RRAGD) gene plays a crucial role in cellular processes. Recently, RRAGD variants found in patients have been implicated in a novel disorder with kidney tubulopathy and dilated cardiomyopathy. Currently, the consequences of RRAGD variants at organismal level is unknown. Therefore, this study investigated the impact of RRAGD variants on cardiac function using zebrafish embryo model. Furthermore, the potential usage of rapamycin, an mTOR inhibitor, as a therapy was assessed in this model. Zebrafish embryos were injected with RRAGD p.S76L and p.P119R cRNA and the resulting heart phenotypes were studied. Our findings reveal that overexpression of RRAGD mutants resulted in decreased ventricular fractional shortening, ejection fraction, and pericardial swelling. In RRAGD S76L-injected embryos, lower survival and heartbeat were observed, while survival was unaffected in RRAGD P119R embryos. These observations were reversible following therapy with the mTOR inhibitor rapamycin. Moreover, no effects on electrolyte homeostasis were observed. Together, these findings indicate a crucial role of RRAGD for cardiac function. In the future, the molecular mechanisms by which RRAGD variants result in cardiac dysfunction, and if the effects of rapamycin are specific for RRAGD-dependent cardiomyopathy should be studied in clinical studies.

The Modulatory Effects and Therapeutic Potential of Cannabidiol in the Gut.

Cannabidiol (CBD) is a major non-psychotropic phytocannabinoid that exists in the Cannabis sativa plant. CBD has been found to act on various receptors, including both cannabinoid and non-cannabinoid receptors. In addition, CBD has antioxidant effects that are independent of receptors. CBD has demonstrated modulatory effects at different organ systems, such as the central nervous system, immune system, and the gastrointestinal system. Due to its broad effects within the body and its safety profile, CBD has become a topic of therapeutic interest. This literature review summarizes previous research findings with regard to the effect of CBD on the gastrointestinal (GI) system, including its effects at the molecular, cellular, organ, and whole-body levels. Both pre-clinical animal studies and human clinical trials are reviewed. The results of the studies included in this literature review suggest that CBD has significant impact on intestinal permeability, the microbiome, immune cells and cytokines. As a result, CBD has been shown to have therapeutic potential for GI disorders such as inflammatory bowel disease (IBD). Furthermore, through interactions with the gut, CBD may also be helpful in the treatment of disorders outside the GI system, such as non-alcoholic liver disease, postmenopausal disorders, epilepsy, and multiple sclerosis. In the future, more mechanistic studies are warranted to elucidate the detailed mechanisms of action of CBD in the gut. In addition, more well-designed clinical trials are needed to explore the full therapeutic potential of CBD on and through the gut.

Effects of mining activities and municipal wastewaters on element accumulation and integrated biomarker responses of the European chub (Squalius cephalus)

This study aimed to determine concentrations of 29 elements in the gills and liver as well as biomarker response in gills, liver, and blood of European chub from Pek River (exposed to long-term mining activities), and to compare these findings with individuals from Ibar River (influenced by emission of treated municipal wastewater) and Kruščica reservoir (source of drinking water) using inductively-coupled plasma optical emission spectrometry (ICP-OES). The metal pollution index (MPI) was also calculated. Supporting analyses for the detection of the municipal wastewater presence at investigated localities included analyses of microbiological indicators (total coliforms and Escherichia coli) of faecal pollution. We have assessed biomarker responses from molecular to organism level using the condition index, comet assay, micronucleus test, oxidative stress parameters, histopathological alterations, and fluorescence spectroscopy parameters. Multibiomarker analysis was summarized by Integrated Biomarker Response v2 (IBRv2). Among these locations, Kruščica exhibited the lowest, whereas the Pek River displayed the highest values for both categories of indicator bacteria. Due to the porphyry copper ores mining, individuals from Pek River had several times higher Cu concentrations in both gills and liver compared to the other localities which was confirmed by biomarker responses and IBRv2 value. On the contrary, fish from Kruščica reservoir were the least affected by elemental pollution which is also confirmed by low MPI and IBRv2 values. Responses of biomarkers correspond to the elemental accumulation in the liver and gills of the Ibar River are positioned between the Pek River and Kruščica reservoir. Of all the biomarkers analyzed in this study, the condition index was the least sensitive. The results of this study showed that fluorescence spectroscopy may be a method for fast screening of structural changes in gills caused by the pollution burden.

Resident Participation in Environmental Governance of Sustainable Tourism in Rural Destination

The rapid development of rural tourism has placed significant pressure on the rural environment, and relying solely on the government and market forces is insufficient for effective governance. It is urgent to integrate endogenous rural forces into environmental governance. The development of social behavior theory offers new insights into exploring sustainable approaches for resident participation in environmental governance in rural tourism areas. This paper, based on the Stimulus-Organism-Response (SOR) theoretical framework from social behavior theory, outlines the entire process of rural tourism residents transitioning from individual stimuli to psychological responses and ultimately to participation in environmental governance. This study combines the Motivation-Opportunity-Ability (MOA) model to analyze the stimulus factors affecting local residents and jointly constructs a path mechanism model for resident participation in environmental governance in rural tourism areas. A total of 462 valid questionnaires were collected through a survey, and the Partial Least Squares Structural Equation Modeling (PLS-SEM) method was used for empirical testing to determine the path coefficients between variables. On this basis, a system dynamics model was constructed to simulate the dynamic evolution of the relationships between variables. This study found the following: (1) In the process of rural tourism, residents’ participation in environmental governance relies on motivational factors at the stimulus level to play a core leading role; opportunity factors act as catalysts, and ability factors serve as auxiliaries; participation willingness at the organism level plays a crucial role. (2) Material pursuit and formal institutions are the strongest single sustainable factors for residents’ participation in environmental governance in rural tourism areas; combinations of variables such as local attachment, governance knowledge, and governance identity have significant effects. It is recommended that future rural tourism leverage government guidance, coordinate interests, and adhere to a collaborative development approach to ensure the sustainable development of the rural tourism environment.

Single Cell Sequencing and Spatial Transcriptomics in Kidney Transplantation

Abstract Purpose of Review Progress in biology and medicine relies on technological advances. The ability to examine gene expression from single cells in tissues with complex architecture is now possible with the advent of single cell sequencing and spatial transcriptomics methods. This review will outline how these methods have been applied to kidney transplantation and their inherent challenges and limitations. Recent Findings The transcriptomes of single cells can now be defined in health and disease allowing for the differences to be investigated. Advanced in situ sequencing and capture methodologies now add spatial resolution to single cell transcriptomics. However, such powerful methods bring challenges, most notably the associated computational burden that is required to make the resulting high-dimensional output data interpretable. A second challenge is that of cost. Thankfully, as with many new technologies’ costs will drop over time making these methods available to more research groups. Summary The sequencing and analysis of gene expression at the single cell level with spatial information retained is now feasible. These methods are major advances for biomedical research and bring huge potential with regards to the better understanding of disease at cellular and organ levels.

Extracellular Matrix and Hepatic Wound Healing before Fibrosis.

The hepatic extracellular matrix (ECM) is most accurately depicted as a dynamic compartment that comprises a diverse range of players that work bidirectionally with hepatic cells to regulate overall homeostasis. Although the classic meaning of the ECM referred to only proteins directly involved in generating the ECM structure, such as collagens, proteoglycans, and glycoproteins, the definition of the ECM is now broader and includes all components associated with this compartment. The ECM is critical in mediating phenotype at the cellular, organ, and even organismal levels. The purpose of this review is to summarize the prevailing mechanisms by which ECM mediates hepatic phenotype and discuss the potential or established role of this compartment in the response to hepatic injury in the context of steatotic liver disease.