Mammalian Tissues Research Articles

Investigating the effects of a FAAH inhibitor in the laterodorsal tegmental nucleus using a new ex vivo mouse preparation

As part of the brainstem reticular activating system, the laterodorsal tegmentum (LDT) is a central player in regulating sleep, arousal, and motivated behaviors including addiction. The neuronal lipids within the LDT that could regulate neuronal activity and LDT signaling are not fully characterized due, in part, to the complication of the presence of fatty acid amide hydrolases (FAAH). To determine the lipids in the LDT, mouse brain slices at different stages of brain development (16, 26 and 66 days) were exposed ex vivo to the FAAH inhibitor oleyl trifluoromethyl ketone (OTMK). Metabolomics and proteomic analyses were conducted on matched samples. Metabolomics analysis revealed differences between OTMK treated and untreated LDT. Furthermore, a distinct phenotype (proteomic profile) as a function of OTMK treatment was observed in LDT from adolescent (66 days) mice indicating an effect of treatment with OTMK at later stages of brain development. Our data indicate that this ex vivo preparation could facilitate screening of different FAAH inhibitors in mammalian tissues, and more importantly, this preparation should allow a deeper characterization of global mass spectrometry-based omics profiles within the LDT.

Ecological and toxicological features of the effect of stable cesium on striated muscle tissue (myocardium and skeletal muscles) of mammals

Recently, the risks of radiation danger have significantly increased. This is connected with the start of a full-scale war against Ukraine and the seizure of the Zaporizhzhya NPP by the Russian Federation, which has the second most powerful arsenal of nuclear weapons in the world. The majority of studies devoted to the effect of radioactive radiation on the human body and animals absolutely do not take into account the ecotoxic effect of the carrier and a chemical element. At the same time, the body does not distinguish between radioactive and non-radioactive isotopes of chemical elements and interacts with them during exchange processes. The aim of the work was to study the influence of stable cesium on the microscopic structure of striated muscle tissue, both skeletal and cardiac ones. The results of our research revealed that intensive accumulation of cesium for 24 days occurred in all the examined organs and tissues. In fact, the most intensive accumulative process appeared to be in the heart by 214.9 times and by 695.3 times with skeletal muscles, with a simultaneous decrease in potassium content by 19.2% and 29.1%, respectively, compared to intact rats. We can talk about the presence of two mechanisms of action of cesium on tissues, including striated muscles: as an antagonist of potassium, which concerns the transport system of the latter and as a toxic element that initiates peroxide processes with subsequent modification of membranes and their potassium channels. Functional disturbances in the work of potassium channels of excitable tissues in the repolarization phase occur in the conditions of an increase in intracellular cesium in striated muscles. These disorders can proceed according to the type of acquired "potassium" channelopathies, which in the myocardium can manifest in the form of rhythm disorders, prolonged QT syndrome, etc. This leads to the appearance of significant violations of the histological structure of the examined tissues. Cesium has pronounced damaging effects on both skeletal and cardiac striated muscle tissue, with morphological changes in skeletal and cardiac striated muscle tissue being generally similar. Dystrophic changes in the nuclei and cytoplasm of cardiomyocytes and striated muscles, signs of impaired blood circulation in the microcirculatory channel with the formation of vascular wall edema, destruction of endothelial cells, various types of hemorrhages and lymphohistiocytic infiltration of the stroma of fibers are the consequences of the toxic effect of cesium on tissues. Thus, the results of changes in the histological structure of striated muscles under the action of cesium chloride obtained by us indicate a fairly high cytocidal activity of stable cesium.

Isomer-Resolved Mass Spectrometry Imaging of Acidic Phospholipids.

The biological functions of lipids are entirely dependent on their molecular structures with even small changes in structure─such as different sites of unsaturation─providing critical markers for changes in the underlying metabolism. Conventional mass spectrometry imaging (MSI) approaches, however, face the twin challenges of mixture and structural complexity and are typically unable to differentiate lipid isomers that differ only in the position(s) of carbon-carbon double bonds. Recent coupling of ozone-induced dissociation (OzID) with matrix-assisted laser desorption/ionization (MALDI)-MSI has demonstrated the potential to map changes in individual double-bond isomers, thus enabling visualization of the modulation in lipid desaturation in adjacent tissue types. This has, to date, only been performed in positive-ion mode due to a generally higher abundance of phosphatidylcholines (PC) in mammalian tissues and the efficient desorption/ionization of this lipid subclass. Many other glycerophospholipids (GPLs), however, are better detected in negative-ion mode as deprotonated anions. Recently, OzID has been implemented on a traveling-wave ion-mobility mass spectrometer (Waters, SYNAPT G2-Si) that provides a 50-fold increase in the rate of the gas-phase reaction between ionized lipids and ozone and a commensurate increase in sensitivity for isomer-resolved mass spectrometry. These gains are exploited here to interrogate the distributions of anionic GPL isomers in biological tissues, covering the subclasses phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylglycerol (PG), and phosphatidic acid (PA). Exploiting both ozone- and collision-induced dissociation in a single acquisition simultaneously identifies sites of unsaturation and acyl chain composition from the same mass spectrum.

Universal DNA methylation age across mammalian tissues

Aging, often considered a result of random cellular damage, can be accurately estimated using DNA methylation profiles, the foundation of pan-tissue epigenetic clocks. Here, we demonstrate the development of universal pan-mammalian clocks, using 11,754 methylation arrays from our Mammalian Methylation Consortium, which encompass 59 tissue types across 185 mammalian species. These predictive models estimate mammalian tissue age with high accuracy (r > 0.96). Age deviations correlate with human mortality risk, mouse somatotropic axis mutations and caloric restriction. We identified specific cytosines with methylation levels that change with age across numerous species. These sites, highly enriched in polycomb repressive complex 2-binding locations, are near genes implicated in mammalian development, cancer, obesity and longevity. Our findings offer new evidence suggesting that aging is evolutionarily conserved and intertwined with developmental processes across all mammals.

Host-microbiome interactions in nicotinamide mononucleotide (NMN) deamidation.

The nicotinamide adenine dinucleotide (NAD+ ) precursor nicotinamide mononucleotide (NMN) is a proposed therapy for age-related disease, whereby it is assumed that NMN is incorporated into NAD+ through the canonical recycling pathway. During oral delivery, NMN is exposed to the gut microbiome, which could modify the NAD+ metabolome through enzyme activities not present in the mammalian host. We show that orally delivered NMN can undergo deamidation and incorporation in mammalian tissue via the de novo pathway, which is reduced in animals treated with antibiotics to ablate the gut microbiome. Antibiotics increased the availability of NAD+ metabolites, suggesting the microbiome could be in competition with the host for dietary NAD+ precursors. These findings highlight new interactions between NMN and the gut microbiome.

Myc beyond Cancer: Regulation of Mammalian Tissue Regeneration.

Myc is one of the most well-known oncogenes driving tumorigenesis in a wide variety of tissues. From the brain to blood, its deregulation derails physiological pathways that grant the correct functioning of the cell. Its action is carried out at the gene expression level, where Myc governs basically every aspect of transcription. Indeed, in addition to its role as a canonical, chromatin-bound transcription factor, Myc rules RNA polymerase II (RNAPII) transcriptional pause-release, elongation and termination and mRNA capping. For this reason, it is evident that minimal perturbations of Myc function mirror malignant cell behavior and, consistently, a large body of literature mainly focuses on Myc malfunctioning. In healthy cells, Myc controls molecular mechanisms involved in pivotal functions, such as cell cycle (and proliferation thereof), apoptosis, metabolism and cell size, angiogenesis, differentiation and stem cell self-renewal. In this latter regard, Myc has been found to also regulate tissue regeneration, a hot topic in the research fields of aging and regenerative medicine. Indeed, Myc appears to have a role in wound healing, in peripheral nerves and in liver, pancreas and even heart recovery. Herein, we discuss the state of the art of Myc's role in tissue regeneration, giving an overview of its potent action beyond cancer.

Batokine in Central Nervous System Diseases.

Brown adipose tissue (BAT) is a special type of fat tissue in mammals and is also a key endocrine organ in the human body. Batokine, the endocrine effector of BAT, plays a neuroprotective role and improves the prognosis by exerting anti-apoptotic and anti-inflammatory effects, as well as by improving vascular endothelial function and other mechanisms in nerve injury diseases. The present article briefly reviewed several types of batokines related to central nervous system (CNS) diseases. Following this, the potential therapeutic value and future research direction of batokines for CNS diseases were chiefly discussed from the aspects of protective mechanism and signaling pathway.

Soft X-rays radiation damage on plunge-frozen and freeze-dried maize roots evaluated by FTIR spectromicroscopy

The radiation damage issue consequent to soft X-rays’ exposure is still an important aspect to be contemplated in soft X-ray microscopy. The work presented here is part of a more extended investigation on the topic and targets the effects of soft X-rays exposure on plant tissues.The aftermaths of soft X-rays’ exposure were evaluated by FTIR spectromicroscopy by comparing irradiated and non-irradiated areas on similar plant structural features, one day and ten days after irradiation, to highlight also a possible time-dependent behavior consequent to air exposure.Our results show partial degradation of lignocellulosic complex and oxidation of cellulose and most probably of hemicellulose; lignin moiety of cell walls in vascular tissue however remains stable.Comparing the current study with our previous works it appears that, as expected, plant tissues seem less prone to soft X-rays radiation damage compared to mammalian cells or tissues.

Structure-property relationships of velar bone tissue from the energy absorbing horncore of bighorn sheep rams.

Velar bone is the material that fills the horncore of bighorn sheep rams. The architectural dimensions of velar bone are orders of magnitude larger than trabecular bone, and velae are more sail-like compared to strut-like trabeculae. Velar bone is important for energy absorption and reduction of brain cavity accelerations during high energy head impacts, but velar bone material properties were previously unknown. It was hypothesized that velar bone tissue would have properties that are beneficial for increased energy absorption at the material level. Solid velar bone beams were tested using dynamic mechanical analysis and three-point bending to quantify mechanical properties. Additionally, the porosity, osteon population density, and mineral content of the solid velar sails were quantified. The velar bone damping factor (∼0.03 - 0.06) and modulus of toughness (3.9 ± 0.4 MJ/m3) were lower than other mammalian cortical bone tissues. The solid bony sails have a bending modulus (8.6 ± 0.5 GPa) that lies within the range of bending moduli values previously reported for individual trabecular struts and cortical bone tissue. The solid velar bone sails had porosity (6.7 ± 0.9 %) and bone mineral content (66 ± 1 %) in the range of cortical bone values. Interestingly, velar sails contained osteons, which are rarely found in trabecular struts. The velar bone osteon population density (5.8 ± 0.9 osteons/mm2) is in the low end of the range of values reported for cortical bone in other mammals. STATEMENT OF SIGNIFICANCE: Bighorn sheep rams sustain high energy head impacts during intraspecific combat without overt signs of brain injury. Previous studies have shown that the bony horncore plays a critical role in energy absorption and reduction of brain cavity accelerations post impact, which has implications for concussion prevention in humans. However, the material properties of the horncore velar bone were previously unknown. This study quantified the material properties and structure-property relationships of the horncore velar bone at the tissue level. Results from this study will improve our understanding of how bighorn sheep mitigate brain injury during head-to-head impacts and may inspire the design of novel materials for energy absorption applications (i.e., helmets materials that reduce concussion occurrence in humans).

Solvent and solid‐phase extraction of per‐ and polyfluoroalkyl substances (PFASs) from mammalian tissues homogenates for UPLC‐MS/MS analysis

Solvent and solid‐phase extraction of per‐ and polyfluoroalkyl substances (PFASs) from mammalian tissues homogenates for UPLC‐MS/MS analysis

Smart biocathodic double signal amplification design empowered self-powered bioplatform ultrasensitive detection of tumor biomarker

MicroRNA-21 (miRNA-21) is widely expressed in various organs and tissues of mammals, including the heart, lungs, brain, pancreas, small intestine, colon, breast, and kidneys, showing high sensitivity and specificity in the diagnosis of multiple types of tumors. It can serve as a new clinical early diagnosis and prognostic biomarker for tumors. This study proposes a self-powered bioplatform, which employs a biological cathode double signal amplification design to construct a high-performance enzyme biofuel cell for the ultra-sensitive detection of tumor biomarker miRNA-21. The biocathodic double signal mechanism involves the catalyzed hairpin self-assembly introduction signal I bilirubin oxidase and a tetrahedral DNA nanostructure that produces a double-helical DNA chain to adsorb signal II electron acceptor [Ru(NH3)6]3+. This approach not only improves the stability of the sensing system, but also synergistically enhances the open circuit voltage. The developed assay exhibits a wide linear range from 0.1 fM to 1000 pM, with a low detection limit of 0.04 fM (S/N = 3). This study provides a strategy for designing biosensors with improved sensitivity and potential application in medical diagnosis field.

Investigation of the chemical composition and anti-inflammatory effects of the crude methanolic extract and fractions of Artocarpus heterophyllus (Fam. Moraceae)

Inflammation is a local response of living tissues in mammals towards injury and is normally linked to the pathogenesis of several diseases. In this study, we investigated the anti-inflammatory activity of the methanol extract and the n-hexane, ethyl acetate and butanol fractions of the Jack fruit leaves. Methanolic cold maceration was carried out to get the extract, followed by liquid-liquid fractionation. Two models, viz. xylene-induced ear edema in mice and egg albumin-induced paw edema in rats were used to evaluate them. The anti-inflammatory activity of the extract and fractions was screened for anti-inflammatory activity using xylene-induced ear edema in mice and egg albumin-induced paw edema in rats. The chemical constituents of the extract and fractions were detected by dereplication using HPLC-DAD. Results showed that for the xylene-induced topical edema model, only the 250µg/ear n-butanol fraction exhibited good anti-inflammatory activity at 21.21% when compared with the piroxicam (positive control) at 32.21%. In the egg albumin-induced paw edema both the methanol extract and n-butanol fraction at 250mg/kg and 500mg/kg showed good anti-inflammatory activity after 2hours post-administration, with maximum effect at 6 hours. Specifically, at 6 hours the methanol extract at 250 mg/kg exhibited 41.21% inhibition; the butanol fraction at the same dose exhibited 43.63% inhibition when compared with piroxicam (500 mg/kg) with 38.39% inhibition. The HPLC-DAD analysis of the extract and fractions revealed the presence of six (6) compounds: catechin (1), septicine (2), 5-isoprunetin 8C glucoside (3), naamine (4), protocatechuic acid (5), and vitexin (6). These compounds have been reported to display anti-inflammatory activity. These findings suggest that the leaf extract possess an anti-inflammatory phytocompounds which authenticates their ethno-medicinal usage claim by local people in different parts of Nigeria.

A Simple, Quick, and Partially Automated Protocol for the Isolation of Single Nuclei from Frozen Mammalian Tissues for Single Nucleus Sequencing.

Single-cell and single-nucleus RNA sequencing have become common laboratory applications due to the wealth of transcriptomic information that they provide. Single nucleus RNA sequencing, particularly, is useful for investigating gene expression in difficult-to-dissociate tissues. Furthermore, this approach is also compatible with frozen (archival) material. Here, we describe a protocol to isolate high-quality single nuclei from frozen mammalian tissues for downstream single nucleus RNA sequencing in a partially-automated manner using commercially available instruments and reagents. Specifically, a robotic dissociator is used to automate and standardize tissue homogenization, followed by an optimized chemical gradient to filter the nuclei. Lastly, we accurately and automatically count the nuclei using an automated fluorescent cell counter. The performance of this protocol is demonstrated on mouse brain, rat kidney, and cynomolgus liver and spleen tissue. This protocol is straightforward, rapid, and readily adaptable to various mammalian tissues without requiring extensive optimization and provides good quality nuclei for downstream single nuclei RNA sequencing.

Autologous blood clots: a natural biomaterial for wound healing

Repair after injury in mammalian tissue involves a complex cascade of events, with the formation of local blood clots being essential for the initial phases of wound healing. As a result, emerging research has sought to harness this biological activity to generate a pro-regenerative biomaterial to speed up wound healing. According to recent studies, “blood clots” created in vitro can be employed as an orthobiologic-based biomaterial for promoting tissue regeneration. Even though such research is still in its early phases, numerous studies show encouraging results that suggest autologous blood clots created in vitro might be a valuable treatment for soft tissue and orthopedic injuries. In this article, we discuss the function of blood clots in physiologic healing, how exogenous material can affect this process, and the most recent clinical research that proposes the use of autologous blood clots as a therapeutically beneficial biomaterial.

Long-Term Treatment with Cannabidiol-Enriched Cannabis Extract Induces Synaptic Changes in the Adolescent Rat Hippocampus.

The endocannabinoid system (eCS) is widely distributed in mammalian tissues and it is classically formed by cannabinoid receptors, endogenous bioactive lipids and its synthesis and degradation enzymes. Due to the modulatory role of eCS in synaptic activity in the Central Nervous System (CNS), phytocannabinoids have been increasingly used for the treatment of neurological disorders, even though little is known in terms of the long-term effect of these treatments on CNS development, mainly in the timeframe that comprises childhood and adolescence. Furthermore, an increased number of clinical trials using full-spectrum Cannabis extracts has been seen, rather than the isolated form of phytocannabinoids, when exploring the therapeutical benefits of the Cannabis plant. Thus, this study aims to evaluate the effect of cannabidiol (CBD)-enriched Cannabis extract on synaptic components in the hippocampus of rats from adolescence to early adulthood (postnatal day 45 to 60). Oral treatment of healthy male Wistar rats with a CBD-enriched Cannabis extract (3 mg/kg/day CBD) during 15 days did not affect food intake and water balance. There was also no negative impact on locomotor behaviour and cognitive performance. However, the hippocampal protein levels of GluA1 and GFAP were reduced in animals treated with the extract, whilst PSD95 levels were increased, which suggests rearrangement of glutamatergic synapses and modulation of astrocytic features. Microglial complexity was reduced in CA1 and CA3 regions, but no alterations in their phagocytic activity have been identified by Iba-1 and LAMP2 co-localization. Collectively, our data suggest that CBD-enriched Cannabis treatment may be safe and well-tolerated in healthy subjects, besides acting as a neuroprotective agent against hippocampal alterations related to the pathogenesis of excitatory and astrogliosis-mediated disorders in CNS.

MOF-mediated histone H4 Lysine 16 acetylation governs mitochondrial and ciliary functions by controlling gene promoters

Histone H4 lysine 16 acetylation (H4K16ac), governed by the histone acetyltransferase MOF, orchestrates gene expression regulation and chromatin interaction. However, the roles of MOF and H4K16ac in controlling cellular function and regulating mammalian tissue development remain unclear. Here we show that conditional deletion of Mof in the skin, but not Kansl1, causes severe defects in the self-renewal of basal epithelial progenitors, epidermal differentiation, and hair follicle growth, resulting in barrier defects and perinatal lethality. MOF-regulated genes are highly enriched for essential functions in the mitochondria and cilia. Genetic deletion of Uqcrq, an essential subunit for the electron transport chain (ETC) Complex III, in the skin, recapitulates the defects in epidermal differentiation and hair follicle growth observed in MOF knockout mouse. Together, this study reveals the requirement of MOF-mediated epigenetic mechanism for regulating mitochondrial and ciliary gene expression and underscores the important function of the MOF/ETC axis for mammalian skin development.

Medium-chain fatty acid oxidation is independent of l-carnitine in liver and kidney but not in heart and skeletal muscle.

Medium-chain fatty acid (MCFA) consumption confers a wide range of health benefits that are highly distinct from long-chain fatty acids (LCFAs). A major difference between the metabolism of LCFAs compared with MCFAs is that mitochondrial LCFA oxidation depends on the carnitine shuttle, whereas MCFA mitochondrial oxidation is not. Although MCFAs are said to range from 6 to 14 carbons long based on physicochemical properties in vitro, the biological cut-off length of acyl chains that can bypass the carnitine shuttle in different mammalian tissues is unknown. To define the range of acyl chain length that can be oxidized in the mitochondria independent of carnitine, we determined the oxidative metabolism of free fatty acids (FFAs) from 6 to 18 carbons long in the liver, kidney, heart, and skeletal muscle. The liver oxidized FFAs 6 to 14 carbons long, whereas the kidney oxidized FFAs from 6 to 10 carbons in length. Heart and skeletal muscle were unable to oxidize FFAs of any chain length. These data show that while the liver and kidney can oxidize MCFAs in the free form, the heart and skeletal muscle require carnitine for the oxidative metabolism of MCFAs. Together these data demonstrate that MCFA oxidation independent of carnitine is tissue-specific.NEW & NOTEWORTHY This work demonstrates that the traditional concept of mitochondrial medium-chain fatty acid oxidation as unregulated and independent of carnitine applies only to liver metabolism, and to kidney to a lesser extent, but not the heart or skeletal muscle. Thus, the benefits of dietary medium-chain fatty acids are set by liver metabolic activity and peripheral tissues are unlikely to receive direct benefits from medium-chain fatty acid metabolism, but rather metabolic byproducts of liver's medium-chain oxidative metabolism.

Anopheline mosquito saliva contains bacteria that are transferred to a mammalian host through blood feeding.

Malaria transmission occurs when Plasmodium sporozoites are transferred from the salivary glands of anopheline mosquitoes to a human host through the injection of saliva. The need for better understanding, as well as novel modes of inhibiting, this key event in transmission has driven intense study of the protein and miRNA content of saliva. Until now the possibility that mosquito saliva may also contain bacteria has remained an open question despite the well documented presence of a rich microbiome in salivary glands. Using both 16S rRNA sequencing and MALDI-TOF approaches, we characterized the composition of the saliva microbiome of An. gambiae and An. stephensi mosquitoes which respectively represent two of the most important vectors for the major malaria-causing parasites P. falciparum and P. vivax. To eliminate the possible detection of non-mosquito-derived bacteria, we used a transgenic, fluorescent strain of one of the identified bacteria, Serratiamarcescens, to infect mosquitoes and detect its presence in mosquito salivary glands as well as its transfer to, and colonization of, mammalian host tissues following a mosquito bite. We also showed that Plasmodium infection modified the mosquito microbiota, increasing the presence of Serratia while diminishing the presence of Elizabethkingia and that both P. berghei and Serratia were transferred to, and colonized mammalian tissues. These data thus document the presence of bacteria in mosquito saliva, their transfer to, and growth in a mammalian host as well as possible interactions with Plasmodium transmission. Together they raise the possible role of mosquitoes as vectors of bacterial infection and the utility of commensal mosquito bacteria for the development of transmission-blocking strategies within a mammalian host.

The Role of S100A6 in Human Diseases: Molecular Mechanisms and Therapeutic Potential.

S100A6, also known as calcyclin, is a low-molecular-weight Ca2+-binding protein from the S100 family that contains two EF-hands. S100A6 is expressed in a variety of mammalian cells and tissues. It is also expressed in lung, colorectal, pancreatic, and liver cancers, as well as other cancers such as melanoma. S100A6 has many molecular functions related to cell proliferation, the cell cycle, cell differentiation, and the cytoskeleton. It is not only involved in tumor invasion, proliferation, and migration, but also the pathogenesis of other non-neoplastic diseases. In this review, we focus on the molecular mechanisms and potential therapeutic targets of S100A6 in tumors, nervous system diseases, leukemia, endometriosis, cardiovascular disease, osteoarthritis, and other related diseases.

Tracing the elimination of reentry spiral waves in defibrillation: Temperature effects

The presence of spiral waves in cardiac tissue causes ventricular fibrillation (VF). Clinical studies have demonstrated that reducing the blood temperature to hypothermia can improve the success rate of electrical defibrillation when VF occurs. In this paper, the Hodgkin-Huxley-style model is used to simulate the effects of temperature and electric shock on the spiral waves present in mammalian tissues, and the phase singularity (PS) identification methods are used to track the wave behaviors in network, finally the Luo-Rudy model is used to test the applicability of our results for cardiac tissues. The results show that temperature modulates the optimal parameters for defibrillation: (1) Arnold's tongue structure shows that an alternating electric field close to the tissue natural frequency (e.g.: cardiac sinus rhythm) allows the nodes to be phase-locked or PS directionally drift at a smaller intensity, thus eliminating spiral waves more effectively than a constant electric field. (2) The average node frequency increases with temperature whether the network is synchronized or present spiral waves, and the statistics show that the robustness of electric defibrillation can be enhanced when the tissue is at moderate hypothermia. (3) More interestingly, we define the defibrillation score (DFS) and find that moderate hypothermia is a balance to improves defibrillation success, which may provide some insights for practical defibrillation studies.