Lithium treatment reverses irradiation-induced changes in rodent neural progenitors and rescues cognition.

Cranial radiotherapy in children has detrimental effects on cognition, mood, and social competence in young cancer survivors. Treatments harnessing hippocampal neurogenesis are currently of great relevance in this context. Lithium, a well-known mood stabilizer, has both neuroprotective, pro-neurogenic as well as antitumor effects, and in the current study we introduced lithium treatment 4 weeks after irradiation. Female mice received a single 4 Gy whole-brain radiation dose on postnatal day (PND) 21 and were randomized to 0.24% Li2CO3 chow or normal chow from PND 49 to 77. Hippocampal neurogenesis was assessed on PND 77, 91, and 105. We found that lithium treatment had a pro-proliferative effect on neural progenitors, but neuronal integration occurred only after it was discontinued. Also, the treatment ameliorated deficits in spatial learning and memory retention observed in irradiated mice. Gene expression profiling and DNA methylation analysis identified two novel factors related to the observed effects, Tppp, associated with microtubule stabilization, and GAD2/65, associated with neuronal signaling. Our results show that lithium treatment reverses irradiation-induced loss of hippocampal neurogenesis and cognitive impairment even when introduced long after the injury. We propose that lithium treatment should be intermittent in order to first make neural progenitors proliferate and then, upon discontinuation, allow them to differentiate. Our findings suggest that pharmacological treatment of cognitive so-called late effects in childhood cancer survivors is possible.

Frequency of the two mating types in the soil under productive and non-productive trees in five French orchards of the Périgord black truffle (Tuber melanosporum Vittad.).

The Périgord black truffle (Tuber melanosporum Vittad.) is an ectomycorrhizal fungus forming edible fructifications. The production of T. melanosporum relies mainly on man-made plantations. T. melanosporum is a heterothallic species requiring the meeting of two partners of opposite mating types to fruit. It is common to have productive and non-productive trees in the same orchard. The aim of our study was to assess the distribution of T. melanosporum mating types in soil under productive and non-productive trees to test whether the presence or absence of one or two mating types could be an indicator of productivity. To achieve this aim, five orchards were selected in various French regions. Soils were harvested under productive and non-productive Quercus pubescens; soil characteristics and the distribution of the mating types in the soil were investigated. No significant differences between productive and non-productive soils according to soil parameters were detected. The total content of T. melanosporum DNA in the soil was significantly higher under productive trees compared with non-productive trees, and it was positively correlated only with soil available phosphorous. Under productive trees, it was more frequent to find both mating types than under non-productive trees. Soils with only one mating type were more frequent under non-productive trees than under productive ones. Moreover, no mating type was detected in the soil of 22% of the non-productive trees. These results suggest that the detection of T. melanosporum mating types in soil could be a tool to optimise the management of truffle orchards (e.g. by spore inoculation).

Aggravated brain injury after neonatal hypoxic ischemia in microglia-depleted mice.

BACKGROUND: Neuroinflammation plays an important role in neonatal hypoxic-ischemic encephalopathy (HIE). Although microglia are largely responsible for injury-induced inflammatory response, they play beneficial roles in both normal and disease states. However, the effects of microglial depletion on neonatal HIE remain unclear. METHODS: Tamoxifen was administered to Cx3cr1CreER/+Rosa26DTA/+ (microglia-depleted model) and Cx3cr1CreER/+Rosa26DTA/- (control) mice at P8 and P9 to assess the effect of microglial depletion. The density of microglia was quantified using Iba-1 staining. Moreover, the proportion of resident microglia after the HI insult was analyzed using flow cytometric analysis. At P10, the HI insult was conducted using the Rice-Vannucci procedure at P10. The infarct size and apoptotic cells were analyzed at P13. Cytokine analyses were performed using quantitative polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA) at P13. RESULTS: At P10, tamoxifen administration induced > 99% microglial depletion in DTA+ mice. Following HI insult, there was persisted microglial depletion over 97% at P13. Compared to male DTA- mice, male DTA+ mice exhibited significantly larger infarct volumes; however, there were no significant differences among females. Moreover, compared to male DTA- mice, male DTA+ mice had a significantly higher density of TUNEL+ cells in the caudoputamen, cerebral cortex, and thalamus. Moreover, compared to female DTA- mice, female DTA+ mice showed a significantly greater number of TUNEL+ cells in the hippocampus and thalamus. Compared to DTA- mice, ELISA revealed significantly lower IL-10 and TGF-ß levels in both male and female DTA+ mice under both normal conditions and after HI (more pronounced). CONCLUSION: We established a microglial depletion model that aggravated neuronal damage and apoptosis after the HI insult, which was predominantly observed in males.

"Hands-On, Minds-On, and Science-Up": A Concept-Based Learning Laboratory With a Taste of Research Experience for an Undergraduate Biomedical Engineering Course.

In this paper, we bridged faculty research expertise with concept-based learning pedagogy to design and implement a unique laboratory experience for biomedical engineering undergraduate students enrolled in the biomechanics of tissues course at the University of Calgary. This laboratory aimed to increase student engagement, facilitate deeper understanding of course content, and provide an opportunity for accelerated undergraduate research through "hands-on," "minds-on," and "science-up" learning components, respectively. The laboratory exercise involves testing aortic tissues using a novel miniaturized planar biaxial machine. This type of machine is normally reserved for use in the context of research. The relevance of the proposed laboratory as a teaching tool was assessed using student feedback. Results indicate an overall valuable and positive learning experience for students.

Ericoid mycorrhizal fungi and their genomes: another side to the mycorrhizal symbiosis?

Contents Summary 1141 I. Introduction 1141 II. The ericoid mycorrhizal lifestyle 1141 III. Lessons from the mycorrhizal fungal genomes 1142 IV. ERM fungi: a discordant voice in the mycorrhizal choir 1143 V. An endophytic niche for ERM fungi 1144 VI. Specialised vs unspecialised mycorrhizal fungi? 1145 VII. Conclusions and perspectives 1145 Acknowledgements 1146 References 1146 SUMMARY: The genome of an organism bears the signature of its lifestyle, and organisms with similar life strategies are expected to share common genomic traits. Indeed, ectomycorrhizal and arbuscular mycorrhizal fungi share some genomic traits, such as the expansion of gene families encoding taxon-specific small secreted proteins, which are candidate effectors in the symbiosis, and a very small repertoire of plant cell wall-degrading enzymes. A large gene family coding for candidate effectors was also revealed in ascomycetous ericoid mycorrhizal (ERM) fungi, but these fungal genomes are characterised by a very high number of genes encoding degradative enzymes, mainly acting on plant cell wall components. We suggest that the genomic signature of ERM fungi mirrors a versatile life strategy, which allows them to occupy several ecological niches.

Comparative genomics and transcriptomics depict ericoid mycorrhizal fungi as versatile saprotrophs and plant mutualists.

Some soil fungi in the Leotiomycetes form ericoid mycorrhizal (ERM) symbioses with Ericaceae. In the harsh habitats in which they occur, ERM plant survival relies on nutrient mobilization from soil organic matter (SOM) by their fungal partners. The characterization of the fungal genetic machinery underpinning both the symbiotic lifestyle and SOM degradation is needed to understand ERM symbiosis functioning and evolution, and its impact on soil carbon (C) turnover. We sequenced the genomes of the ERM fungi Meliniomyces bicolor, M. variabilis, Oidiodendron maius and Rhizoscyphus ericae, and compared their gene repertoires with those of fungi with different lifestyles (ecto- and orchid mycorrhiza, endophytes, saprotrophs, pathogens). We also identified fungal transcripts induced in symbiosis. The ERM fungal gene contents for polysaccharide-degrading enzymes, lipases, proteases and enzymes involved in secondary metabolism are closer to those of saprotrophs and pathogens than to those of ectomycorrhizal symbionts. The fungal genes most highly upregulated in symbiosis are those coding for fungal and plant cell wall-degrading enzymes (CWDEs), lipases, proteases, transporters and mycorrhiza-induced small secreted proteins (MiSSPs). The ERM fungal gene repertoire reveals a capacity for a dual saprotrophic and biotrophic lifestyle. This may reflect an incomplete transition from saprotrophy to the mycorrhizal habit, or a versatile life strategy similar to fungal endophytes.

Three-dimensional thoracic aorta principal strain analysis from routine ECG-gated computerized tomography: feasibility in patients undergoing transcatheter aortic valve replacement.

BACKGROUND: Functional impairment of the aorta is a recognized complication of aortic and aortic valve disease. Aortic strain measurement provides effective quantification of mechanical aortic function, and 3-dimenional (3D) approaches may be desirable for serial evaluation. Computerized tomographic angiography (CTA) is routinely performed for various clinical indications, and offers the unique potential to study 3D aortic deformation. We sought to investigate the feasibility of performing 3D aortic strain analysis in a candidate population of patients undergoing transcatheter aortic valve replacement (TAVR). METHODS: Twenty-one patients with severe aortic valve stenosis (AS) referred for TAVR underwent ECG-gated CTA and echocardiography. CTA images were analyzed using a 3D feature-tracking based technique to construct a dynamic aortic mesh model to perform peak principal strain amplitude (PPSA) analysis. Segmental strain values were correlated against clinical, hemodynamic and echocardiographic variables. Reproducibility analysis was performed. RESULTS: The mean patient age was 81±6 years. Mean left ventricular ejection fraction was 52±14%, aortic valve area (AVA) 0.6±0.3 cm2 and mean AS pressure gradient (MG) 44±11 mmHg. CTA-based 3D PPSA analysis was feasible in all subjects. Mean PPSA values for the global thoracic aorta, ascending aorta, aortic arch and descending aorta segments were 6.5±3.0, 10.2±6.0, 6.1±2.9 and 3.3±1.7%, respectively. 3D PSSA values demonstrated significantly more impairment with measures of worsening AS severity, including AVA and MG for the global thoracic aorta and ascending segment (p<0.001 for all). 3D PSSA was independently associated with AVA by multivariable modelling. Coefficients of variation for intra- and inter-observer variability were 5.8 and 7.2%, respectively. CONCLUSIONS: Three-dimensional aortic PPSA analysis is clinically feasible from routine ECG-gated CTA. Appropriate reductions in PSSA were identified with increasing AS hemodynamic severity. Expanded study of 3D aortic PSSA for patients with various forms of aortic disease is warranted.

Collagen structural alterations contribute to stiffening of tissue after split-thickness skin grafting.

The gold standard treatment for full thickness injuries of the skin is autologous split-thickness skin grafting. This involves harvesting the epidermis and superficial dermis from healthy skin and transplanting it onto the prepared wound bed. The donor site regenerates spontaneously, but the appendages and cellular components from the dermal layer are excluded from the graft. As a result, the new tissue is inferior; the healed graft site is dry/itchy, has decreased elasticity, increased fragility, and altered sensory function. Because this dermal layer is composed of collagen and other extracellular matrix proteins, the aim was to characterize the changes in the dermal collagen after split thickness grafting that could contribute to a deficit in functionality. This will serve as a baseline for future studies designed to improve skin function using pharmacological or cell-based therapies for skin repair. A xenograft model whereby human split-thickness grafts were implanted into full-thickness defects on immunocompromised (athymic Nu/Nu) mice was used. The grafts were harvested 4 and 8 weeks later. The collagen microstructure was assessed with second harmonic generation with dual-photon microscopy and light polarization analysis. Collagen fiber stiffness and engagement stretch were estimated by fitting the results of biaxial mechanical tensile tests to a histo-mechanical constitutive model. The stiffness of the collagen fibril-proteoglycan complex increased from 682 ± 226 kPa/sr to 1016 ± 324 kPa/sr between 4 and 8 weeks postgrafting. At the microstructural level there were significant decreases in both thickness of collagen fibers (3.60 ± 0.34 µm vs. 2.10 ± 0.27 µm) and waviness ratio (2.04 ± 0.17 vs. 1.43 ± 0.08) of the collagen fibers postgrafting. The decrease of the macroscopic engagement stretch from 1.19 ± 0.11 to 1.09 ± 0.08 over time postgrafting mirrored the decrease in waviness measured at the microscopic level. This suggested that the integrity of the collagen fibers was compromised and contributed to the functional deficit of the skin postgrafting.

Model systems to unravel the molecular mechanisms of heavy metal tolerance in the ericoid mycorrhizal symbiosis.

Ericoid mycorrhizal plants dominate in harsh environments where nutrient-poor, acidic soil conditions result in a higher availability of potentially toxic metals. Although metal-tolerant plant species and ecotypes are known in the Ericaceae, metal tolerance in these plants has been mainly attributed to their association with ericoid mycorrhizal fungi. The mechanisms underlying plant protection by the fungal symbiont are poorly understood, whereas some insights have been achieved regarding the molecular mechanisms of heavy metal tolerance in the fungal symbiont. This review will briefly introduce the general features of heavy metal tolerance in mycorrhizal fungi and will then focus on the use of "omics" approaches and heterologous expression in model organisms to reveal the molecular bases of fungal response to heavy metals. Functional complementation in Saccharomyces cerevisiae has allowed the identification of several ericoid mycorrhizal fungi genes (i.e., antioxidant enzymes, metal transporters, and DNA damage repair proteins) that may contribute to metal tolerance in a metal-tolerant ericoid Oidiodendron maius isolate. Although a powerful system, the use of the yeast complementation assay to study metal tolerance in mycorrhizal symbioses has limitations. Thus, O. maius has been developed as a model system to study heavy metal tolerance mechanisms in mycorrhizal fungi, thanks to its high metal tolerance, easy handling and in vitro mycorrhization, stable genetic transformation, genomics, transcriptomic and proteomic resources.

New mutualistic fungal endophytes isolated from poplar roots display high metal tolerance.

This study aimed to isolate, identify, and characterise metal-tolerant fungi colonising poplar roots at a metal-contaminated phytoremediation site. Poplar roots were colonised by arbuscular mycorrhizal, ectomycorrhizal, and endophytic fungi, and the species were determined by ITS molecular analyses. Eight different isolates were successfully isolated into pure culture. Three isolates belonging to the Helotiales (P02, P06) and the Serendipita vermifera species (P04) were highly tolerant to metals (Cd, Zn, Pb, and Cu) compared to the mycorrhizal Hebeloma isolates. The three isolates degraded complex carbohydrates, such as xylan and cellulose, indicating that they could partially degrade root cell walls and penetrate into cells. This hypothesis was confirmed by further in vitro re-synthesis experiments, which showed that the three isolates colonised root tissues of poplar plantlets whereas two of them formed microsclerotia-like structures. Taken together, these results suggest an endophytic lifestyle of these isolates. This is the first evidence of S. vermifera as a root endophyte of poplar. A new endophytic putative species belonging to the Helotiales and closely related to Leohumicola is also reported. Interestingly, and when compared to mock-inoculated plants, both P06 and P04 isolates increased the number of root tips of inoculated poplar plantlets in vitro. Moreover, the S. vermifera P04 isolate also increased the shoot biomass. The results are discussed in relation to the potential use of endophytic strains for tree-based phytoremediation of metal-contaminated sites.

Brain Maturation as a Fundamental Factor in Immune-Neurovascular Interactions in Stroke.

Injuries in the developing brain cause significant long-term neurological deficits. Emerging clinical and preclinical data have demonstrated that the pathophysiology of neonatal and childhood stroke share similar mechanisms that regulate brain damage, but also have distinct molecular signatures and cellular pathways. The focus of this review is on two different diseases-neonatal and childhood stroke-with emphasis on similarities and distinctions identified thus far in rodent models of these diseases. This includes the susceptibility of distinct cell types to brain injury with particular emphasis on the role of resident and peripheral immune populations in modulating stroke outcome. Furthermore, we discuss some of the most recent and relevant findings in relation to the immune-neurovascular crosstalk and how the influence of inflammatory mediators is dependent on specific brain maturation stages. Finally, we comment on the current state of treatments geared toward inducing neuroprotection and promoting brain repair after injury and highlight that future prophylactic and therapeutic strategies for stroke should be age-specific and consider gender differences in order to achieve optimal translational success.

Insight in Quinazoline-based HDAC Inhibitors as Anti-cancer Agents.

Cancer remains a primary cause of death globally, and effective treatments are still limited. While chemotherapy has notably enhanced survival rates, it brings about numerous side effects. Consequently, the ongoing challenge persists in developing potent anti-cancer agents with minimal toxicity. The versatile nature of the quinazoline moiety has positioned it as a pivotal component in the development of various antitumor agents, showcasing its promising role in innovative cancer therapeutics. This concise review aims to reveal the potential of quinazolines in creating anticancer medications that target histone deacetylases (HDACs).

An alternative conformation of the N-terminal loop of human dihydroorotate dehydrogenase drives binding to a potent antiproliferative agent.

Over the years, human dihydroorotate dehydrogenase (hDHODH), which is a key player in the de novo pyrimidine-biosynthesis pathway, has been targeted in the treatment of several conditions, including autoimmune disorders and acute myelogenous leukaemia, as well as in host-targeted antiviral therapy. A molecular exploration of its inhibitor-binding behaviours yielded promising candidates for innovative drug design. A detailed description of the enzymatic pharmacophore drove the decoration of well-established inhibitory scaffolds, thus gaining further in vitro and in vivo efficacy. In the present work, using X-ray crystallography, an atypical rearrangement was identified in the binding pose of a potent inhibitor characterized by a polar pyridine-based moiety (compound 18). The crystal structure shows that upon binding compound 18 the dynamics of a protein loop involved in a gating mechanism at the cofactor-binding site is modulated by the presence of three water molecules, thus fine-tuning the polarity/hydrophobicity of the binding pocket. These solvent molecules are engaged in the formation of a hydrogen-bond mesh in which one of them establishes a direct contact with the pyridine moiety of compound 18, thus paving the way for a reappraisal of the inhibition of hDHODH. Using an integrated approach, the thermodynamics of such a modulation is described by means of isothermal titration calorimetry coupled with molecular modelling. These structural insights will guide future drug design to obtain a finer Kd/logD7.4 balance and identify membrane-permeable molecules with a drug-like profile in terms of water solubility.

Inflammatory, metabolic, and sex-dependent gene-regulatory dynamics of microglia and macrophages in neonatal hippocampus after hypoxia-ischemia.

Neonatal hypoxia-ischemia (HI) is a major cause of perinatal death and long-term disabilities worldwide. Post-ischemic neuroinflammation plays a pivotal role in HI pathophysiology. In the present study, we investigated the temporal dynamics of microglia (CX3CR1GFP/+) and infiltrating macrophages (CCR2RFP/+) in the hippocampi of mice subjected to HI at postnatal day 9. Using inflammatory pathway and transcription factor (TF) analyses, we identified a distinct post-ischemic response in CCR2RFP/+ cells characterized by differential gene expression in sensome, homeostatic, matrisome, lipid metabolic, and inflammatory molecular signatures. Three days after injury, transcriptomic signatures of CX3CR1GFP/+ and CCR2RFP/+ cells isolated from hippocampi showed a partial convergence. Interestingly, microglia-specific genes in CX3CR1GFP/+ cells showed a sexual dimorphism, where expression returned to control levels in males but not in females during the experimental time frame. These results highlight the importance of further investigations on metabolic rewiring to pave the way for future interventions in asphyxiated neonates.

OmZnT1 and OmFET, two metal transporters from the metal-tolerant strain Zn of the ericoid mycorrhizal fungus Oidiodendron maius, confer zinc tolerance in yeast.

Two full-length cDNAs (OmZnT1 and OmFET) encoding membrane transporters were identified by yeast functional screening in the heavy metal tolerant ericoid mycorrhizal isolate Oidiodendron maius Zn. OmZnT1 belongs to the Zn-Type subfamily of the cation diffusion facilitators, whereas OmFET belongs to the family of iron permeases. Their properties were investigated in yeast by functional complementation of mutants affected in metal uptake and metal tolerance. Heterologous expression of OmZnT1 and OmFET in a Zn-sensitive yeast mutant restored the wild-type phenotype. Additionally, OmZnT1 expression also restored cobalt tolerance in a Co-sensitive mutant. A GFP fusion protein revealed that OmZnT1 was targeted to the endoplasmic reticulum membrane, a result consistent with a function for OmZnT1 in metal sequestration. Similarly to other iron permeases, OmFET-GFP was localized on the plasma membrane. OmFET restored the growth of uptake-defective strains for iron and zinc. Zinc-sensitive yeast mutants expressing OmFET specifically accumulated magnesium, as compared to cells transformed with the empty vector. We suggest that OmFET may counteract zinc toxicity by increasing entry of magnesium into the cell.

Morphological and mechanical examination of the atrial 'intima'.

AIMS: To examine morphology and mechanical properties of the atrial 'intima', which we defined as the tissue interposed between atrial endocardium and myocardium, in patients without known cardiovascular disease. METHODS AND RESULTS: Post-mortem right and left atrial tissue was obtained from male infants (<1 year, n = 4), children (10-19 years, n = 4), and adults (58-69 years, n = 7). Using light microscopy and an ocular micrometer, atrial intimal (AIT) thickness was measured. Intimal collagen bundle thickness was measured using electron microscopy. Passive atrial wall stiffness was measured using a planar biaxial testing device. Among infants, left AIT (0.2 ± 0.2 mm) and right (0.2 ± 0.1 mm) AIT were not significantly different (P = 0.84). Among children, left AIT (0.6 ± 0.2 mm) was significantly greater than right (0.2 ± 0.1 mm) AIT (P = 0.03), and left AIT was marginally greater than in infants (P = 0.07). Among adults, with the exception of the appendage region, left AIT (1.0 ± 0.2 mm) was markedly greater than right AIT (0.3 ± 0.1 mm; P < 0.05), and left AIT was significantly greater than that in other age groups (P < 0.05). There were no differences in right AIT among age groups. Left intimal collagen bundle thickness was greater in adults (0.0512 ± 0.0056 µm) than infants (0.0432 ± 0.0071 µm) or children (0.0435 ± 0.0013 µm), and bundles were less organized. Wall stiffness was attributable primarily to the intima (1245 ± 132, vs. 260 ± 45 N/m(2) for the remaining atrial wall). CONCLUSION: The left atrial intima, but not the right, thickens with age, becomes more disorganized ultrastructurally, and is responsible for the majority of atrial wall stiffness.

A feature-based morphing methodology for computationally modeled biological structures applied to left atrial fiber directions.

To properly simulate the behavior of biological structures through computer modeling, there exists a need to describe parameters that vary locally. These parameters can be obtained either from literature or from experimental data and they are often assigned to regions in the model as lumped values. Furthermore, parameter values may be obtained on a representative case and may not be available for each specific modeled organ. We describe a semiautomated technique to assign detailed maps of local tissue properties to a computational model of a biological structure. We applied the method to the left atrium of the heart. The orientation of myocytes in the tissue as obtained from histologic analysis was transferred to the 3D model of a porcine left atrium. Finite element method (FEM) dynamic simulations were performed by using an isotropic, neo-Hookean, constitutive model first, then adding an anisotropic, cardiomyocyte oriented, Fung-type component. Results showed higher stresses for the anisotropic material model corresponding to lower stretches in the cardiomyocyte directions. The same methodology can be applied to transfer any map of parameters onto a discretized finite element model.

AI-powered assessment of biomarkers for growth prediction of abdominal aortic aneurysms.

Objective: The purpose of this study was to employ biomechanics-based biomarkers to locally characterize abdominal aortic aneurysm (AAA) tissue and investigate their relation to local aortic growth by means of an artificial intelligence model. Methods: The study focused on a population of 36 patients with AAAs undergoing serial monitoring with electrocardiogram-gated multiphase computed tomography angiography acquisitions. The geometries of the aortic lumen and wall were reconstructed from the baseline scans and used for the baseline assessment of regional aortic weakness with three functional biomarkers, time-averaged wall-shear stress, in vivo principal strain, and intra-luminal thrombus thickness. The biomarkers were encoded as regional averages on axial and circumferential sections perpendicularly to the aortic centerline. Local diametric growth was obtained as difference in diameter between baseline and follow-up at the level of each axial section. An artificial intelligence model was developed to predict accelerated aneurysmal growth with the Extra Trees algorithm used as a binary classifier where the positive class represented regions that grew more than 2.5 mm/year. Additional clinical biomarkers, such as maximum aortic diameter at baseline, were also investigated as predictors of growth. Results: The area under the curve for the constructed receiver operating characteristic curve for the Extra Trees classifier showed a very good performance in predicting relevant aortic growth (area under the curve = 0.92), with the three biomechanics-based functional biomarkers being objectively selected as the main predictors of growth. Conclusions: The use of features based on the functional and local characterization of the aortic tissue resulted in a superior performance in terms of growth prediction when compared with models based on geometrical assessments. With rapid growth linked to increasing risk for patients with AAAs, the ability to access functional information related to tissue weakening and disease progression at baseline has the potential to support early clinical decisions and improve disease management.