Loss Of Plasticity Research Articles

Codonopsis pilosula polysaccharide attenuates Aβ toxicity and cognitive defects in APP/PS1 mice.

Codonopsis pilosula Polysaccharides (CPPs), a traditional Chinese medicine used for thousands of years, is a potential neuroprotective polysaccharide via a relatively poorly understood mechanism. We previously reported that CPPs attenuated tau pathology in hTau transfected mice and therefore in the current work investigated the effect of CPPs on Aβ toxicity and cognitive defects in APP/PS1 mice model. It was found that one-month intragastric administration of CPPs significantly ameliorated cognitive defects in APP/PS1 mice. In addition, CPPs treatment mitigated the loss of the synaptic plasticity and increased the synaptic proteins including synaptotagmin and PSD95. The expression of Aβ42 and Aβ40 was remarkably decreased in the hippocampus of APP/PS1 mice after CPPs treatment. We also found that CPPs coincubation significantly reduced the amount of APPβ and Aβ42 expression in cells. Intriguingly, the activity of BACE1 was decreased following CPPs treatment in both the hippocampus of APP/PS1 mice and in vitro experiments. Collectively, these results indicated that CPPs attenuated Aβ pathology in APP/PS1 mice, and down-regulating BACE1 might be the underlaying mechanism which could be a therapeutic target for alleviating cognitive defects in AD pathology.

Long-term durability assessment of PVC-P waterproofing geomembranes through laboratory tests

Waterproofing heavily influences the operation and maintenance costs of underground structures. Currently, the most commonly used technology for tunnel waterproofing is plasticized polyvinyl chloride (PVC-P) geomembranes. However, not much is known about the long-term durability of these geomembranes, especially in relation to the long expected lifespan of new tunnels (i.e. 100–150 years). Therefore, in this paper, the durability of two commercially available PVC-P geomembranes is studied with the help of a specifically designed accelerated ageing device in addition to mechanical and absorption tests. The degradation resulting from plasticizer loss is extrapolated to the long term, and a threshold value for the end-of-life of the PVC-P geomembrane is estimated from the mechanical tests.

Metabolic Communication and Healthy Aging: Where Should We Focus Our Energy?

Aging is associated with a loss of metabolic homeostasis and plasticity, which is causally linked to multiple age-onset pathologies. The majority of the interventions-genetic, dietary, and pharmacological-that have been found to slow aging and protect against age-related disease in various organisms do so by targeting central metabolic pathways. However, targeting metabolic pathways chronically and ubiquitously makes it difficult to define the downstream effects responsible for lifespan extension and often results in negative effects on growth and health, limiting therapeutic potential. Insight into how metabolic signals are relayed between tissues, cells, and organelles opens up new avenues to target metabolic regulators locally rather than globally for healthy aging. In this review, we discuss the pro-longevity effects of targeting metabolic pathways in specific tissues and how these interventions communicate with distal cells to modulate aging. These studies may be crucial in designing interventions that promote longevity without negative health consequences.

Strength and deformability of different types of geosynthetic barriers

Introduction. Water engineering facilities utilize geosynthetic barriers of various materials, but the most common are polyvinylchloride (PVC) and polyethylene (PE) considered to be the most efficient. However, the utilization of thermoset geosynthetics (PVC and PE) as watertight elements of the geosynthetic barriers of earth dams has also a number of drawbacks. In the course of time, PVC can lose its properties due to plasticizer loss, whereas PE is susceptible to cracking at thermal impacts. That is why it is necessary to consider geosynthetic barriers of other types as alternatives. For this purpose, we studied the deformability and the strength of geosynthetic barriers of thermoset material (chlorosulfonated polyethylene — CSPE), as well as of a rubber-type synthetic material (ethylene-propylene diene monomer — EPDM). Materials and methods. Sample testing of geosynthetic barrier materials for monoaxial extension was conducted by means of a tensile-testing machine, which records during the experiment the changes of the tensile force and the longitudinal extension. The changes of the cross-section area of the samples were measured by means of a digital side caliper. Results. It was determined that of the considered geosynthetic barrier types (PE, PVC, CSPE and EPDM) the strongest one is the reinforced CSPE type and the most deformable is the EPDM barrier. Reinforced CSPE geosynthetic barrier has a tensile strength above 100 MPa, it is approximately 5 times higher than that of PVC and PE barriers. By deformability, reinforced CSPE barrier is similar to PE barriers. An EPDM geosynthetic barrier has the linear deformation modulus around 1 MPa, it is capable of a multiple extension without loss of strength. Conclusions. By strength vs. deformability ratio, EPDM geosynthetic barriers can compete with PVC barriers.

Sustainable Micro and Nano Additives for Controlling the Migration of a Biobased Plasticizer from PLA-Based Flexible Films.

Plasticized poly(lactic acid) (PLA)/poly(butylene succinate) (PBS) blend-based films containing chitin nanofibrils (CN) and calcium carbonate were prepared by extrusion and compression molding. On the basis of previous studies, processability was controlled by the use of a few percent of a commercial acrylic copolymer acting as melt strength enhancer and calcium carbonate. Furthermore, acetyl n-tributyl citrate (ATBC), a renewable and biodegradable plasticizer (notoriously adopted in PLA based products) was added to facilitate not only the processability but also to increase the mechanical flexibility and toughness. However, during the storage of these films, a partial loss of plasticizer was observed. The consequence of this is not only correlated to the change of the mechanical properties making the films more rigid but also to the crystallization and development of surficial oiliness. The effect of the addition of calcium carbonate (nanometric and micrometric) and natural nanofibers (chitin nanofibrils) to reduce/control the plasticizer migration was investigated. The prediction of plasticizer migration from the films’ core to the external surface was carried out and the diffusion coefficients, obtained by regression of the experimental migration data plotted as the square root of time, were evaluated for different blends compositions. The results of the diffusion coefficients, obtained thanks to migration tests, showed that the CN can slow the plasticizer migration. However, the best result was achieved with micrometric calcium carbonate while nanometric calcium carbonate results were less effective due to favoring of some bio polyesters’ chain scission. The use of both micrometric calcium carbonate and CN was counterproductive due to the agglomeration phenomena that were observed.

Caveolin-1 regulates medium spiny neuron structural and functional plasticity.

Caveolin-1 (CAV1) is a structural protein critical for spatial organization of neuronal signaling molecules. Whether CAV1 is required for long-lasting neuronal plasticity remains unknown. We sought to examine the effects of CAV1 knockout (KO) on functional plasticity and hypothesized that CAV1 deficiency would impact drug-induced long-term plasticity in the nucleus accumbens (NAc). We first examined cell morphology of NAc medium spiny neurons in a striatal/cortical co-culture system before moving in vivo to study effects of CAV1 KO on cocaine-induced plasticity. Whole-cell patch-clamp recordings were performed to determine effects of chronic cocaine (15 mg/kg) on medium spiny neuron excitability. To test for deficits in behavioral plasticity, we examined the effect of CAV1 KO on locomotor sensitization. Disruption of CAV1 expression leads to baseline differences in medium spiny neuron (MSN) structural morphology, such that MSNs derived from CAV1 KO animals have increased dendritic arborization when cultured with cortical neurons. The effect was dependent on phospholipase C and cell-type intrinsic loss of CAV1. Slice recordings of nucleus accumbens shell MSNs revealed that CAV1 deficiency produces a loss of neuronal plasticity. Specifically, cocaine-induced firing rate depression was absent in CAV1 KO animals, whereas baseline electrophysiological properties were similar. This was reflected by a loss of cocaine-mediated behavioral sensitization in CAV1 KO animals, with unaffected baseline locomotor responsiveness. This study highlights a critical role for nucleus accumbens CAV1 in plasticity related to the administration of drugs of abuse.

LTP-like cortical plasticity predicts conversion to dementia in patients with memory impairment

BackgroundNew diagnostic criteria consider Alzheimer’s disease (AD) as a clinico-biological entity identifiable in vivo on the presence of specific patterns of CSF biomarkers. ObjectiveHere we used transcranial magnetic stimulation to investigate the mechanisms of cortical plasticity and sensory-motor integration in patients with hippocampal-type memory impairment admitted for the first time in the memory clinic stratified according to CSF biomarkers profile. MethodsSeventy-three patients were recruited and divided in three groups according to the new diagnostic criteria: 1) Mild Cognitive Impaired (MCI) patients (n = 21); Prodromal AD (PROAD) patients (n = 24); AD with manifest dementia (ADD) patients (n = 28). At time of recruitment all patients underwent CSF sampling for diagnostic purposes. Repetitive and paired-pulse transcranial magnetic stimulation protocols were performed to investigate LTP-like and LTD-like cortical plasticity, short intracortical inhibition (SICI) and short afferent inhibition (SAI). Patients were the followed up during three years to monitor the clinical progression or the conversion to dementia. ResultsMCI patients showed a moderate but significant impairment of LTP-like cortical plasticity, while ADD and PROAD groups showed a more severe loss of LTP-like cortical plasticity. No differences were observed for LTD-like cortical plasticity, SICI and SAI protocols. Kaplan-Meyer analyses showed that PROAD and MCI patients converting to dementia had weaker LTP-like plasticity at time of first evaluation. ConclusionLTP-like cortical plasticity could be a novel biomarker to predict the clinical progression to dementia in patients with memory impairment at prodromal stages of AD identifiable with the new diagnostic criteria based on CSF biomarkers.

Exploring Sphingolipid Implications in Neurodegeneration.

Over the past decade, it was found that relatively simple sphingolipids, such as ceramide, sphingosine, sphingosine-1-phosphate, and glucosylceramide play important roles in neuronal functions by regulating rates of neuronal growth and differentiation. Homeostasis of membrane sphingolipids in neurons and myelin is essential to prevent the loss of synaptic plasticity, cell death and neurodegeneration. In our review we summarize data about significant brain cell alterations of sphingolipids in different neurodegenerative diseases such as Alzheimer's disease, Parkinson disease, Amyotrophic Lateral Sclerosis, Gaucher's, Farber's diseases, etc. We reported results obtained in brain tissue from both animals in which diseases were induced and humans in autopsy samples. Moreover, attention was paid on sphingolipids in biofluids, liquor and blood, from patients. In Alzheimer's disease sphingolipids are involved in the processing and aggregation of β-amyloid and in the transmission of the cytotoxic signal β-amyloid and TNFα-induced. Recently, the gangliosides metabolism in transgenic animals and the relationship between blood sphingolipids changes and cognitive impairment in Alzheimer's disease patients have been intensively studied. Numerous experiments have highlighted the involvement of ceramide and monohexosylceramide metabolism in the pathophysiology of the sporadic forms of Parkinson's disease. Moreover, gene mutations of the glucocerebrosidase enzyme were considered as responsible for Parkinson's disease via transition of the monomeric form of α-synuclein to an oligomeric, aggregated toxic form. Disturbances in the metabolism of ceramides were also associated with the appearance of Lewy's bodies. Changes in sphingolipid metabolism were found as a manifestation of Amyotrophic Lateral Sclerosis, both sporadic and family forms, and affected the rate of disease development. Currently, fingolimod (FTY720), a sphingosine-1-phosphate receptor modulator, is the only drug undergoing clinical trials of phase II safety for the treatment of Amyotrophic Lateral Sclerosis. The use of sphingolipids as new diagnostic markers and as targets for innovative therapeutic strategies in different neurodegenerative disorders has been included.

Nutrition-responsive gene expression and the developmental evolution of insect polyphenism.

Nutrition-responsive development is a ubiquitous and highly diversified example of phenotypic plasticity, yet its underlying molecular and developmental mechanisms and modes of evolutionary diversification remain poorly understood. We measured genome-wide transcription in three closely related species of horned beetles exhibiting strikingly diverse degrees of nutrition responsiveness in the development of male weaponry. We show that (1) counts of differentially expressed genes between low- and high-nutritional backgrounds mirror species-specific degrees of morphological nutrition responsiveness; (2) evolutionary exaggeration of morphological responsiveness is underlain by both amplification of ancestral nutrition-responsive gene expression and recruitment of formerly low nutritionally responsive genes; and (3) secondary loss of morphological responsiveness to nutrition coincides with a dramatic reduction in gene expression plasticity. Our results further implicate genetic accommodation of ancestrally high variability of gene expression plasticity in both exaggeration and loss of nutritional plasticity, yet reject a major role of taxon-restricted genes in the developmental regulation and evolution of nutritional plasticity.

Cigarette smoke exposure has region-specific effects on GDAP1 expression in mouse hippocampus

Early detection markers for substance use disorders are urgently needed. Recently, an association between the methylation of Ganglioside-induced differentiation-associated protein 1 (GDAP1) and alcohol addiction was found in a US and German population. In this study, we investigate whether GDAP1 expression might be affected by cigarette smoke as well and thus might be a marker of substance addiction in general. 11 adult female C57BL/6 J mice (6 wildtype and 5 lacking the NO-sensitive guanylyl cyclase1 (NO-GC1 KO)) were exposed to cigarette smoke over a period of 5 weeks, their brains immunohistochemically stained and compared to 11 non exposed mice (5 WT and 6 KO). The deletion of NO-GC1 results in a complete loss of synaptic plasticity, therefore, addiction-related alterations might become more obvious. Co-staining of anti-GDAP1 and DAPI revealed protein in the stratum granulare of the hippocampus. Three randomized frames for dentate gyrus (DG) and three for Cornu Ammonis region 1 (CA1) were used to count GDAP1. Cigarette smoke exposure significantly influenced GDAP1 expression depending on the hippocampal region but was not influenced by guanyl cyclase. In conclusion, cigarette smoke exposure alone had an effect on GDAP1 amount in both regions. Therewith, GDAP1might be a biomarker for substance addiction in general.

Neuroprotective effects of cerium oxide nanoparticles on experimental stress-induced depression in male rats

Introduction: Inflammation and oxidative/nitrative stress induced by chronic psychosocial or physical stress play fundamental roles in the pathogenesis of depression, and lead to the loss of neurotrophic support, decreased neurogenesis and synaptic plasticity in the hippocampus. Findings have shown inhibition of inflammation waterfall may offer new approaches to the treatment of depression, especially for patients with treatment-resistant depression (TRD). Cerium Oxide (CeO2) is the oxide form of cerium, a rare earth element in the lanthanide series. Cerium oxide nanoparticles (CeO2NPs) are potent regenerative antioxidant and anti-inflammatory agents. We evaluated the neuroprotective and the neuronal plasticity activities of CeO2NPs in the stress-induced model of depression. Methods: We have analyzed the effects of single-dose intrahippocampal and intracerebroventricular (ICV) injections of CeO2NPs treatment on immobility behavior (forced swimming test), hippocampal IL-6 and malondialdehyde (Elisa), cell survival (Nissl staining), dentate gyrus BrdU-positive cells (Immunohistofluorescence) and growth associated protein-43 (GAp-43) expression (Immunohistofluorescence) of the hippocampal CA3 region. Results: In our study, single-dose CeO2NPs treatment could effectively suppress inflammatory and oxidative markers level induced by unpredictable chronic mild stress (UCMS) and improved immobility behavior, increased hippocampal cell proliferation, cell survival and neurite outgrowth, and thereby, prevented the progression of hippocampal destruction and dysfunction. Conclusion: This study demonstrates a new prospect for the treatment of depression. Our study gives us the confidence to continue the investigation of CeO2 NPs as a novel therapy, especially, for TRD that characterized by chronic and persistent inflammation.

Comparative Study on Hydrogen Embrittlement Susceptibility in Heat-Affected Zone of TP321 Stainless Steel

TP321 stainless steel is widely used in hydrogenation refining pipes owing to its excellent performance of creep stress resistance and high-temperature resistance. In this study, thermal simulation tests were carried out on the welding heat-affected zone (HAZ) of TP321 stainless steel at temperatures of 1300 °C, 1100 °C, and 850°C using a Gleeble 3800 testing machine. Slow strain tensile tests were conducted under the condition of electrolytic hydrogen charging (EHC) and the metallographic microstructure of cracks as well as the morphology of fractures were analyzed in detail. The result shows that hydrogen can change the fracture mode of tensile specimen and the cracks initiated from and near the specimen surface after EHC. Hydrogen significantly decreases the plastic deformation capability of HAZ in TP321 stainless steel. The reduction of area after the fracture decreases by 58%, 41%, and 45% for HAZ at 1300 °C, 1100 °C, and 850 °C, respectively. The existence of δ ferrite was considered to be the main reason for the aggravation of hydrogen-induced plasticity loss.

Chronic Presence of Oligomeric Aβ Differentially Modulates Spine Parameters in the Hippocampus and Cortex of Mice With Low APP Transgene Expression.

Alzheimer’s disease is regarded as a synaptopathy with a long presymptomatic phase. Soluble, oligomeric amyloid-β (Aβ) is thought to play a causative role in this disease, which eventually leads to cognitive decline. However, most animal studies have employed mice expressing high levels of the Aβ precursor protein (APP) transgene to drive pathology. Here, to understand how the principal neurons in different brain regions cope with moderate, chronically present levels of Aβ, we employed transgenic mice expressing equal levels of mouse and human APP carrying a combination of three familial AD (FAD)-linked mutations (Swedish, Dutch, and London), that develop plaques only in old age. We analyzed dendritic spine parameters in hippocampal and cortical brain regions after targeted expression of EGFP to allow high-resolution imaging, followed by algorithm-based evaluation of mice of both sexes from adolescence to old age. We report that Aβ species gradually accumulated throughout the life of APPSDL mice, but not the oligomeric forms, and that the amount of membrane-associated oligomers decreased at the onset of plaque formation. We observed an age-dependent loss of thin spines under most conditions as an indicator of a loss of synaptic plasticity in older mice. We further found that hippocampal pyramidal neurons respond to increased Aβ levels by lowering spine density and shifting spine morphology, which reached significance in the CA1 subfield. In contrast, the spine density in cortical pyramidal neurons of APPSDL mice was unchanged. We also observed an increase in the protein levels of PSD-95 and Arc in the hippocampus and cortex, respectively. Our data demonstrated that increased concentrations of Aβ have diverse effects on dendritic spines in the brain and suggest that hippocampal and cortical neurons have different adaptive and compensatory capacity during their lifetime. Our data also indicated that spine morphology differs between sexes in a region-specific manner.

High post-anthesis temperature effects on bread wheat (Triticum aestivum L.) grain transcriptome during early grain-filling

BackgroundHigh post-anthesis (p.a) temperatures reduce mature grain weights in wheat and other cereals. However, the causes of this reduction are not entirely known. Control of grain expansion by the maternally derived pericarp of the grain has previously been suggested, although this interaction has not been investigated under high p.a. temperatures. Down-regulation of pericarp localised genes that regulate cell wall expansion under high p.a. temperatures may limit expansion of the encapsulated endosperm due to a loss of plasticity in the pericarp, reducing mature grain weight. Here the effect of high p.a. temperatures on the transcriptome of the pericarp and endosperm of the wheat grain during early grain-filling was investigated via RNA-Seq and is discussed alongside grain moisture dynamics during early grain development and mature grain weight.ResultsHigh p.a. temperatures applied from 6-days after anthesis (daa) and until 18daa reduced the grain’s ability to accumulate water, with total grain moisture and percentage grain moisture content being significantly reduced from 14daa onwards. Mature grain weight was also significantly reduced by the same high p.a. temperatures applied from 6daa for 4-days or more, in a separate experiment. Comparison of our RNA-Seq data from whole grains, with existing data sets from isolated pericarp and endosperm tissues enabled the identification of subsets of genes whose expression was significantly affected by high p.a. temperature and predominantly expressed in either tissue. Hierarchical clustering and gene ontology analysis resulted in the identification of a number of genes implicated in the regulation of cell wall expansion, predominantly expressed in the pericarp and significantly down-regulated under high p.a. temperatures, including endoglucanase, xyloglucan endotransglycosylases and a β-expansin. An over-representation of genes involved in the ‘cuticle development’ functional pathway that were expressed in the pericarp and affected by high p.a. temperatures was also observed.ConclusionsHigh p.a. temperature induced down-regulation of genes involved in regulating pericarp cell wall expansion. This concomitant down-regulation with a reduction in total grain moisture content and grain weight following the same treatment period, adds support to the theory that high p.a. temperatures may cause a reduction in mature grain weight as result of decreased pericarp cell wall expansion.

Low-dose X-ray imaging may increase the risk of neurodegenerative diseases

The hypothesis presented here explores the possibility that X-ray imaging commonly used in dental practices may be a shared risk factor for sporadic dementias and motor-neuron diseases. As the evidence will suggest, the brain is ill-equipped to manage the intrusion of low-dose ionizing radiation (IR) beyond that which is naturally occurring. When the brain’s antioxidant defenses are overwhelmed by IR, it produces an abundance of reactive oxygen species (ROS) that can lead to oxidative stress, mitochondrial dysfunction, loss of synaptic plasticity, altered neuronal structure and microvascular impairment that have been identified as early signs of neurodegeneration in Alzheimer’s disease, Parkinson’s, amyotrophic lateral sclerosis, vascular dementia and other diseases that progressively damage the brain and central nervous system. Although genes play a role in all outcomes, the focus here will be on the non-genetic processes at work. Common assumptions regarding the risks of low-dose IR will be addressed, such as: 1) comparing rapid, repeated bursts of man-made IR sent exclusively into the head to equivalent amounts of head-to-toe background IR over longer periods of time; 2) whether epidemiological studies that dismiss concerns regarding low-dose IR due to lack of evidence it causes cancer, heritable mutations or shortened life spans also apply to neurodegeneration; and 3) why even radiation-resistant neurons can be severely impacted by IR exposure, due to IR-induced injury to the processes they need to function. Also considered will be the difficulty of distinguishing the effects of dental X-ray exposure from similarly low amounts of background IR and where to find the evidence that they may, in fact, be responsible for neurodegeneration. Finally, the long-standing belief that whatever risks are inherent in dental radiography must be undertaken for the sake of oral health is challenged on two counts: 1) while dentists continue to drape their patients in lead-lined aprons, the most effective IR safety precautions are often ignored; and 2) there is an alternative dental imaging technology that does not use IR. While the thrust of this article will be on dental radiation and will touch on how age, gender, X-ray equipment and protocols may increase risk, chiropractic radiographs also will be considered because they focus exclusively on the central nervous system. If X-ray imaging is found to be associated with neurodegeneration, the risk-versus-benefit must be reevaluated, every means of reducing exposure implemented and imaging protocols revised.

Effect of sulfated galactans from red seaweed Gracilaria fisheri on extracellular matrix production in human dermal fibroblast

Fibroblasts are derived from mesenchymal stem cells that play an important role in produce the extracellular matrix (ECM). ECM creates a tissue microenvironment for regulation of cell signaling. The loss of ECM plasticity is associated with several pathologies, especially those involving chronic inflammation, therefore, the ECM represents a potential therapeutic target for certain conditions. Sulfated galactans was extracted from red seaweed Gracilaria fisheri that previous studies have been reported about its activities on anti‐oxidant, anti‐tumour, anti‐melanogenesis, and anti‐bacteria. Therefore, the aim of this study was to investigate the effect of potential of sulfated galactans from red seaweed G. fisheri on enhancing the ECM production. Cytotoxicity was tested following determination of ECM‐related gene and protein expression on human dermal fibroblast (HDF) by quantitative RT‐PCR and enzyme‐linked immunosorbent assay (ELISA), respectively. The result showed that sulfated galactans not effects to survival rate of HDF. Additionally, we found that sulfated galactans was able to modulate ECM production by regulating the collagen, elastin, hyaluronic acid, MMP‐1, and TIMP production. The present results demonstrating the effects of a natural multi‐component compound on ECM composition, highlighted the possibility of pharmacologically modulating ECM molecules. The recovery and the maintenance of ECM homeostasis might be considered as a potential therapeutic goal to ameliorate pathological conditions.Support or Funding InformationThis work was financially supported by Navamindradhiraj University Research Fund.

Maturity-dependent cartilage cell plasticity and sensitivity to external perturbation

ObjectiveArticular cartilage undergoes biological and morphological changes throughout maturation. The prevalence of osteoarthritis in the aged population suggests that maturation predisposes cartilage to degradation and/or impaired regeneration, but this process is not fully understood. Therefore, the objective of this study was to characterize the cellular and genetic profile of cartilage, as well as biological plasticity in response to mechanical and culture time stimuli, as a function of animal maturity. Methods/designPorcine articular cartilage explants were harvested from stifle joints of immature (2–4 weeks), adolescent (5–6 months), and mature (1–5 years) animals. Half of all samples were subjected to a single compressive mechanical load. Loaded samples were paired with unloaded controls for downstream analyses. Expression of cartilage progenitor cell markers CD105, CD44, and CD29 were determined via flow cytometry. Expression of matrix synthesis genes Col1, Col2, Col10, ACAN, and SOX9 were determined via qPCR. Tissue morphology and matrix content were examined histologically. Post-loading assays were performed immediately and following 7 days in culture. ResultsCD105 and CD29 expression decreased with maturity, while CD44 expression was upregulated in cartilage from mature animals. Expression of matrix synthesis genes were generally upregulated in cartilage from mature animals, and adolescent animals showed the lowest expression of several matrix synthesizing genes. Culture time and mechanical loading analyses revealed greater plasticity to mechanical loading and culture time in cartilage from younger animals. Histology confirmed distinct structural and biochemical profiles across maturity. ConclusionThis study demonstrates differential, nonlinear expression of chondroprogenitor markers and matrix synthesis genes as a function of cartilage maturity, as well as loss of biological plasticity in aged tissue. These findings have likely implications for age-related loss of regeneration and osteoarthritis progression.

Highly dense cellulose acetate specimens with superior mechanical properties produced by fused filament fabrication

The extrusion based Fused Filament Fabrication (FFF) enables to build up components of thermoplastic materials by using a layer-by-layer strategy. Typically, the layered structure results in an internal porosity limiting the mechanical performance. In this work, using a commercial 3D-printer specimens of the polymer cellulose acetate (CA) were produced having porosities as low as 1–3 %. Quasi-static tensile tests demonstrate that these highly dense specimens reach similar or even higher strength values in comparison to their injection molded counterparts. The influence of the temperature management was studied by using different hot-ends, namely a standard one and a novel hot-end having a modified heat-block. Using the latter one, a homogeneous internal structure was achieved as observed with scanning electron microscopy (SEM). The very low porosities can be assigned to the elevated specimen temperature maintained during printing which is slightly above the materials glass transition temperature. Processing of CA at such temperatures leads to a slight loss of plasticizer as proved with NMR spectroscopy. This favorable change in material composition during printing additionally contributes to the mechanical strength of the test specimens. We conclude that the porosity is the key parameter (which has to be measured and minimized) to realize FFF components with good mechanical performance. Whereas we demonstrated FFF for the first time for CA as an industrially relevant thermoplastic material, our results can be easily applied to amorphous polymers in general.

Partial Metal Ion Saturation of C2 Domains Primes Synaptotagmin 1-Membrane Interactions

Synaptotagmin 1 (Syt1) is an integral membrane protein whose phospholipid-binding tandem C2 domains, C2A and C2B, act as Ca2+ sensors of neurotransmitter release. Our objective was to understand the role of individual metal-ion binding sites of these domains in the membrane association process. We used Pb2+, a structural and functional surrogate of Ca2+, to generate the protein states with well-defined protein-metal ion stoichiometry. NMR experiments revealed that binding of one divalent metal ion per C2 domain results in loss of conformational plasticity of the loop regions, potentially pre-organizing them for additional metal-ion and membrane-binding events. In C2A, a divalent metal ion in site 1 is sufficient to drive its weak association with phosphatidylserine-containing membranes, whereas in C2B, it enhances the interactions with the signaling lipid phosphatidylinositol-4,5-bisphosphate. In full-length Syt1, both Pb2+-complexed C2 domains associate with phosphatidylserine-containing membranes. Electron paramagnetic resonance experiments show that the extent of membrane insertion correlates with the occupancy of the C2 metal ion sites. Together, our results indicate that upon partial metal ion saturation of the intra-loop region, Syt1 adopts a dynamic, partially membrane-bound state. The properties of this state, such as conformationally restricted loop regions and positioning of C2 domains in close proximity to anionic lipid headgroups, “prime” Syt1 for cooperative binding of a full complement of metal ions and deeper membrane insertion.

Half-Sib Lines of Pedunculate Oak (Quercus robur L.) Respond Differently to Drought Through Biometrical, Anatomical and Physiological Traits

Quercus robur L. is one of the most valued tree species of deciduous temperate forests. However, in the last decade, serious oak declines and loss of adaptation plasticity have been reported throughout Europe as a consequence of drought. Therefore, the aim of the present study was to define the adaptation potential of five Q. robur half-sib lines from the UNESCO Biosphere Reserve Mura-Drava-Danube to drought, using physiological, anatomical and biometrical traits. Half-sib lines that exhibited drought tolerance had particular suites of trait expression regarding biometrical traits (seedling height, root length, root to shoot ratio of dry mass and specific leaf area), leaf stomatal traits (stomatal density per mm2, stomata guard cell length and width, stomatal aperture length and width) and leaf structural traits (adaxial epidermis thickness, palisade parenchyma thickness, spongy parenchyma thickness, lamina thickness). All of the observed parameters of chlorophyll a fluorescence were shown to be good indicators of short-term and severe drought. For the selection of drought-tolerant half-sib lines, all studied chlorophyll a fluorescence parameters associated with the heat dissipation of light energy (coefficient of non-photochemical quenching, quantum yield of regulated energy dissipation, Stern-Volmer type non-photochemical fluorescence quenching) and one parameter related to photochemical dissipation of light energy (effective quantum yield (efficiency) of PS II photochemistry) were proven to be suitable. On the other hand, the coefficient of photochemical quenching, coefficient of photochemical fluorescence quenching assuming interconnected photosystem II antennae and electron transport rate were not suitable for distinguishing the different responses of the studied half-sib lines under drought. The importance of results of the present study is in the selection of drought-tolerant Q. robur half-sib lines for future reforestation programs, particularly in protected areas with sensitive forest management and restricted activities for mitigation of the adverse effects of climate changes.