Insight into the structure-elastic property relationship of calcium silicate glasses: a multi-length scale approach.

Based on a combination of molecular dynamics simulations, and Raman and Brillouin light scattering spectroscopies, we investigate the structure and elastic properties relationship in an archetypical calcium silicate glass system. From molecular dynamics and Raman spectroscopy, we show that the atomic structure at the short and intermediate length scales is made up of long polymerized silicate chains, which adjusts itself by closing the Si-O-Si angles and leaving more space to [CaO]n edge shared polyhedra to strengthen the glass. Using Brillouin spectroscopy, we observe an increase of elastic constants of the glass with the calcium content, as the cohesion of the glass structure is enhanced through an increase of the binding between the cross-linked calcium-silicate frameworks. This result, albeit being simple in its nature, illustrates for the first time the implication of the calcium framework in the elastic behavior of the glass and will contribute substantially to the understanding of the composition-structure-property relationships in multi-component industrial glasses.

SHIRPA as a Neurological Screening Battery in Mice.

The SmithKline, Harwell, Imperial College, Royal Hospital, Phenotype Assessment (SHIRPA) is a rapid battery of tests comprising 42 measurements of motor activity, coordination, postural control, muscle tone, autonomic functions, and emotional reactivity, as well as reflexes dependent on visual, auditory, and tactile modalities. Individual scores in SHIRPA are sensitive in detecting phenotypes of several experimental models of neural disease, especially cerebellar degeneration and Alzheimer disease, and combined subscores have been useful in estimating the impact of vascular anomalies and exposure to infectious agents. In cerebellar degeneration, weak forelimb grip, impaired wire maneuver and air righting, and negative geotaxis appear as prevalent features. Most of the measures in the battery are susceptible to change after gene modifications or physiological alterations. SHIRPA can be used both in adult mice and mice in the preweaning period to screen for sensorimotor function and emotional reactivity, not selective attention or memory. © 2021 Wiley Periodicals LLC Basic Protocol: Step-by-step procedure for SHIRPA.

Betacellulin regulates schwann cell proliferation and myelin formation in the injured mouse peripheral nerve.

When a nerve fiber is cut or crushed, the axon segment that is separated from the soma degenerates distal from the injury in a process termed Wallerian degeneration (WD). C57BL/6OlaHsd-WldS (WldS ) mutant mice exhibit significant delays in WD. This results in considerably delayed Schwann cell and macrophage responses and thus in impaired nerve regenerations. In our previous work, thousands of genes were screened by DNA microarrays and over 700 transcripts were found to be differentially expressed in the injured sciatic nerve of WldS compared with wild-type (WT) mice. One of these transcripts, betacellulin (Btc), was selected for further analysis since it has yet to be characterized in the nervous system, despite being known as a ligand of the ErbB receptor family. We show that Btc mRNA is strongly upregulated in immature and dedifferentiated Sox2+ Schwann cells located in the sciatic nerve distal stump of WT mice, but not WldS mutants. Transgenic mice ubiquitously overexpressing Btc (Tg-Btc) have increased numbers of Schmidt-Lantermann incisures compared with WT mice, as revealed by Coherent anti-Stokes Raman scattering (CARS). Tg-Btc mice also have faster nerve conduction velocity. Finally, we found that deficiency in Btc reduces the proliferation of myelinating Schwann cells after sciatic nerve injury, while Btc overexpression induces Schwann cell proliferation and improves recovery of locomotor function. Taken together, these results suggest a novel regulatory role of Btc in axon-Schwann cell interactions involved in myelin formation and nerve repair. GLIA 2017 GLIA 2017;65:657-669.

Tissue-Plasminogen Activator Attenuates Alzheimer's Disease-Related Pathology Development in APPswe/PS1 Mice.

Alzheimer's disease (AD) is the leading cause of dementia among elderly population. AD is characterized by the accumulation of beta-amyloid (Aß) peptides, which aggregate over time to form amyloid plaques in the brain. Reducing soluble Aß levels and consequently amyloid plaques constitute an attractive therapeutic avenue to, at least, stabilize AD pathogenesis. The brain possesses several mechanisms involved in controlling cerebral Aß levels, among which are the tissue-plasminogen activator (t-PA)/plasmin system and microglia. However, these mechanisms are impaired and ineffective in AD. Here we show that the systemic chronic administration of recombinant t-PA (Activase rt-PA) attenuates AD-related pathology in APPswe/PS1 transgenic mice by reducing cerebral Aß levels and improving the cognitive function of treated mice. Interestingly, these effects do not appear to be mediated by rt-PA-induced plasmin and matrix metalloproteinases 2/9 activation. We observed that rt-PA essentially mediated a slight transient increase in the frequency of patrolling monocytes in the circulation and stimulated microglia in the brain to adopt a neuroprotective phenotype, both of which contribute to Aß elimination. Our study unraveled a new role of rt-PA in maintaining the phagocytic capacity of microglia without exacerbating the inflammatory response and therefore might constitute an interesting approach to stimulate the key populations of cells involved in Aß clearance from the brain.

Neurobehavioral Anomalies in the Pitx3/ak Murine Model of Parkinson's Disease and MPTP.

Pitx3/ak null mutants are characterized by basal ganglia pathology in a manner resembling Parkinson's disease (PD), with decline in substantia nigra cell numbers as well as striatal tyrosine hydroxylase expression. Although young adult Pitx3/ak mutants were deficient in motor coordination tests, they were more active than non-transgenic controls in the open-field, unlike PD-related bradykinesia. On the SHIRPA primary screen, the mutants displayed body tremor, hyperactivity in the viewing jar, anomalies in eye morphology as well as a higher degree of hindlimb clasping and myoclonic jumping. Increased hindlimb clasping time and rotorod deficits seen in mutants were also exhibited by mice injected with MPTP, indicating an influence of dopamine on these behaviors.

Effect of charge polydispersity and charge residence time on the dynamics of a micellar system.

We use dynamic light scattering to investigate the effects of charge polydispersity and charge residence time on the dynamics of a micellar system. While in the corresponding uncharged system only one exponential relaxation is observed, two relaxation modes are seen when charging the micelles by adding charged co-surfactant molecules with a long residence time. We attribute the existence of these two relaxation modes to the combined effect of size polydispersity and charge polydispersity, i.e. frozen fluctuations of the number of charges per micelle. Further support to this scenario is provided by control experiments on a similar charged system, but where the charge residence time is short compared to the time scales probed by dynamic light scattering. Here, charge polydispersity is effectively suppressed due to the rapid exchange of charged molecules between micelles and only one single relaxation mode is seen, thereby demonstrating the key role of frozen charge fluctuations in the complex dynamics of our micellar system.

Memory formation and retention are affected in adult miR-132/212 knockout mice.

The miR-132/212 family is thought to play an important role in neural function and plasticity, while its misregulation has been observed in various neurodegenerative disorders. In this study, we analyzed 6-month-old miR-132/212 knockout mice in a battery of cognitive and non-cognitive behavioral tests. No significant changes were observed in reflexes and basic sensorimotor functions as determined by the SHIRPA primary screen. Accordingly, miR-132/212 knockout mice did not differ from wild-type controls in general locomotor activity in an open-field test. Furthermore, no significant changes of anxiety were measured in an elevated plus maze task. However, the mutant mice showed retention phase defects in a novel object recognition test and in the T-water maze. Moreover, the learning and probe phases in the Barnes maze were clearly altered in knockout mice when compared to controls. Finally, changes in BDNF, CREB, and MeCP2 were identified in the miR-132/212-deficient mice, providing a potential mechanism for promoting memory loss. Taken together, these results further strengthen the role of miR-132/212 in memory formation and retention, and shed light on the potential consequences of its deregulation in neurodegenerative diseases.

Sensorimotor skills in Fxn KO/Mck mutants deficient for frataxin in muscle.

Friedreich ataxia is the most common autosomal recessive disorder of the cerebellum, causing degeneration of spinal sensory neurons and spinocerebellar tracts. The disease is caused by severely reduced levels of frataxin, a mitochondrial protein involved in iron metabolism. An experimental model has been generated by crossing mice homozygous for a conditional allele of the Fxn gene with mice heterozygous for a deleted exon 4 of Fxn carrying a tissue-specific Cre transgene under control of the muscle creatine kinase promoter. Relative to wild-type, Fxn null mutants were impaired on tests of motor coordination comprising horizontal bar, vertical pole, and the rotorod as well as displaying gait anomalies and the hindlimb clasping response. The Fxn KO/Mck model reproduces some key features of patients with Friedreich ataxia and provides an opportunity of ameliorating their symptoms with experimental therapies.

GPR84 deficiency reduces microgliosis, but accelerates dendritic degeneration and cognitive decline in a mouse model of Alzheimer's disease.

Microglia surrounds the amyloid plaques that form in the brains of patients with Alzheimer's disease (AD), but their role is controversial. Under inflammatory conditions, these cells can express GPR84, an orphan receptor whose pathophysiological role is unknown. Here, we report that GPR84 is upregulated in microglia of APP/PS1 transgenic mice, a model of AD. Without GPR84, these mice display both accelerated cognitive decline and a reduced number of microglia, especially in areas surrounding plaques. The lack of GPR84 affects neither plaque formation nor hippocampal neurogenesis, but promotes dendritic degeneration. Furthermore, GPR84 does not influence the clinical progression of other diseases in which its expression has been reported, i.e., experimental autoimmune encephalomyelitis (EAE) and endotoxic shock. We conclude that GPR84 plays a beneficial role in amyloid pathology by acting as a sensor for a yet unknown ligand that promotes microglia recruitment, a response affecting dendritic degeneration and required to prevent further cognitive decline.

Motor activity in young APPswe + PS1/A246E bigenic mice as a predicting variable for memory decline.

Reports of individuality in rodent species have been a subject of debate in pharmacology and other fields. In the current study, APPswe + PS1/A246E bigenic mice with Alzheimer's-like pathogenesis and wild-type controls were subdivided at 3 months of age into low, intermediate, and high responders in open-field activity. The mice were then evaluated longitudinally at 3 and 9 months for object recognition. Irrespective of genotype, mice with a high level of motor activity had better scores in object recognition. However, a significant correlation was established between open-field activity measured at 3 months of age and recognition memory measured at 9 months of age in the bigenic group only. These results indicate that motor activity in young mice with amyloid neuropathology may serve as a predicting variable for cognitive dysfunction in more mature mice.

Interactions between microemulsion droplets decorated with hydrophobically modified polymers: a small-angle neutron scattering study.

The shape and interactions between microemulsion droplets (R = 8.2 nm, polydispersity 20%) either decorated with PEO modified with a single hydrophobic end function (PEO-m: C12H25 - (EO)n, M(PEO) = 5.2 kg/mol), or with telechelic polymers of twice the mass (PEO-2m: C12H25 - (EO)2n - C12H25, M(PEO) = 10.4 kg/mol) have been studied by small-angle neutron scattering (SANS). The results as a function of droplet and polymer concentration have been compared to the reference case of the bare microemulsion which was shown to be unchanged using Porod representations. The interactions between bare and decorated droplets have been analyzed using the structure factor S(q), at first in a model-free way based on its low-q limit S(q → 0). This analysis provides clear evidence on the concentration-dependent repulsive or attractive nature of the contributions to the pair droplet-droplet pair potential of the polymers. Model pair potentials describing the steric repulsions and attractions by copolymer bridging are used to describe the low-q behavior of the structure factor based on an integral equation approach, giving an estimate of the range and amplitude of the potentials. Moreover, they provide an explanation for the observed transient clustering in terms of a shallow minimum of the total potential, as they establish the respective repulsive and attractive contributions of the polymer molecules.

An AAV9 coding for frataxin clearly improved the symptoms and prolonged the life of Friedreich ataxia mouse models.

Friedreich ataxia (FRDA) is a genetic disease due to increased repeats of the GAA trinucleotide in intron 1 of the frataxin gene. This mutation leads to a reduced expression of frataxin. We have produced an adeno-associated virus (AAV)9 coding for human frataxin (AAV9-hFXN). This AAV was delivered by intraperitoneal (IP) injection to young conditionally knockout mice in which the frataxin gene had been knocked-out in some tissues during embryogenesis by breeding them with mice expressing the Cre recombinase gene under the muscle creatine kinase (MCK) or the neuron-specific enolase (NSE) promoter. In the first part of the study, different doses of virus were tested from 6 × 10(11) v.p. to 6 × 10(9) v.p. in NSE-cre mice and all leading to an increase in life spent of the mice. The higher and the lower dose were also tested in MCK-cre mice. A single administration of the AAV9-hFXN at 6 × 10(11) v.p. more than doubled the life of these mice. In fact the MCK-cre mice treated with the AAV9-hFXN were sacrificed for further molecular investigations at the age of 29 weeks without apparent symptoms. Echography analysis of the heart function clearly indicated that the cardiac systolic function was better preserved in the mice that received 6 × 10(11) v.p. of AAV9-hFXN. The human frataxin protein was detected by ELISA in the heart, brain, muscles, kidney, and liver with the higher dose of virus in both mouse models. Thus, gene therapy with an AAV9-hFXN is a potential treatment of FRDA.

Protein phosphatase 2A family members (PP2A and PP6) associate with U1 snRNP and the spliceosome during pre-mRNA splicing.

Protein phosphorylation and dephosphorylation are both important for multiple steps in the splicing pathway. Members of the PP1 and PP2A subfamilies of phospho-serine/threonine phosphatases play essential but redundant roles in the second step of the splicing reaction. PP6, a member of the PP2A subfamily, is the mammalian homolog of yeast Sit4p and ppe1, which are involved in cell cycle regulation; however, the involvement of PP6 in the splicing pathway remains unclear. Here we show that PP2A family members physically associate with the spliceosome throughout the splicing reaction. PP2A holoenzyme and PP6 were found stably associated with U1 snRNP. Together our findings indicate that these phosphatases regulate splicing catalysis involving U1 snRNP and suggest an important evolutionary conserved role of PP2A family phosphatases in pre-mRNA splicing.

Toll-like receptor 4 stimulation with the detoxified ligand monophosphoryl lipid A improves Alzheimer's disease-related pathology.

Alzheimer's disease (AD) is the most common cause of dementia worldwide. The pathogenesis of this neurodegenerative disease, currently without curative treatment, is associated with the accumulation of amyloid ß (Aß) in brain parenchyma and cerebral vasculature. AD patients are unable to clear this toxic peptide, leading to Aß accumulation in their brains and, presumably, the pathology associated with this devastating disease. Compounds that stimulate the immune system to clear Aß may therefore have great therapeutic potential in AD patients. Monophosphoryl lipid A (MPL) is an LPS-derived Toll-like receptor 4 agonist that exhibits unique immunomodulatory properties at doses that are nonpyrogenic. We show here that repeated systemic injections of MPL, but not LPS, significantly improved AD-related pathology in APP(swe)/PS1 mice. MPL treatment led to a significant reduction in Aß load in the brain of these mice, as well as enhanced cognitive function. MPL induced a potent phagocytic response by microglia while triggering a moderate inflammatory reaction. Our data suggest that the Toll-like receptor 4 agonist MPL may be a treatment for AD.

The effects of subchronic d-serine on left-right discrimination learning, social interaction, and exploratory activity in APPswe/PS1 mice.

Glutamatergic neurotransmission is crucially involved in memory and cognition and severely affected patients with Alzheimer's disease. Modulation of NMDA receptors with agonists may reverse their late-stage symptoms. The effects of subchronic treatment of the NMDA receptor agonist, d-serine, were evaluated in APPswe/PS1 mutant mice harboring Aß plaques in brain, regarding spatial discrimination learning, open-field activity, and social interaction in a three-chambered apparatus. d-serine (50mg/kg, i.p.) was superior to placebo in mutant mice during the reversal phase of left-right discrimination learning without affecting acquisition. The drug caused weaker effects in counteracting open-field hyperactivity and low social interaction with an unfamiliar stimulus mouse. These results indicate a favorable action of an NMDA receptor agonist on reversal learning in transgenic mice with amyloid pathology.

Cognitive and non-cognitive behaviors in the triple transgenic mouse model of Alzheimer's disease expressing mutated APP, PS1, and Mapt (3xTg-AD).

3xTg-AD mutant mice are characterized by parenchymal Aß plaques and neurofibrillary tangles resembling those found in patients with Alzheimer's disease. The mutants were compared with non-transgenic controls in sensorimotor and learning tests. 3xTg-AD mutants were deficient in T-maze reversal, object recognition, and passive avoidance learning. In addition, the mutants showed hypoactivity in two open-field tests, fewer fecal boli in an observation jar, and reduced enclosed arm entries and head-dipping in the elevated plus-maze. On the contrary, the mutants did not differ from controls in pain thresholds, nest-building, and various reflexes determined by the SHIRPA primary screen and were even better on the rotorod test of motor coordination.

Functional recovery after peripheral nerve injury is dependent on the pro-inflammatory cytokines IL-1ß and TNF: implications for neuropathic pain.

IL-1ß and TNF are potential targets in the management of neuropathic pain after injury. However, the importance of the IL-1 and TNF systems for peripheral nerve regeneration and the mechanisms by which these cytokines mediate effects are to be fully elucidated. Here, we demonstrate that mRNA and protein levels of IL-1ß and TNF are rapidly upregulated in the injured mouse sciatic nerve. Mice lacking both IL-1ß and TNF, or both IL-1 type 1 receptor (IL-1R1) and TNF type 1 receptor (TNFR1), showed reduced nociceptive sensitivity (mechanical allodynia) compared with wild-type littermates after injury. Microinjecting recombinant IL-1ß or TNF at the site of sciatic nerve injury in IL-1ß- and TNF-knock-out mice restored mechanical pain thresholds back to levels observed in injured wild-type mice. Importantly, recovery of sciatic nerve function was impaired in IL-1ß-, TNF-, and IL-1ß/TNF-knock-out mice. Notably, the infiltration of neutrophils was almost completely prevented in the sciatic nerve distal stump of mice lacking both IL-1R1 and TNFR1. Systemic treatment of mice with an anti-Ly6G antibody to deplete neutrophils, cells that play an essential role in the genesis of neuropathic pain, did not affect recovery of neurological function and peripheral axon regeneration. Together, these results suggest that targeting specific IL-1ß/TNF-dependent responses, such as neutrophil infiltration, is a better therapeutic strategy for treatment of neuropathic pain after peripheral nerve injury than complete blockage of cytokine production.

Sensorimotor and cognitive functions in a SOD1(G37R) transgenic mouse model of amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a neurological disorder involving degeneration of motor neurons in brain and spinal cord, leading to progressive atrophy of skeletal muscles and, ultimately, paralysis and death. Copper-mediated oxidative damage is proposed to play a critical role in the pathogenesis of Cu/Zn superoxide dismutase (SOD1) - linked hereditary amyotrophic lateral sclerosis. To understand more clearly the pathogenesis of sensorimotor dysfunction and to find the most appropriate methods for early detection of symptoms and for monitoring them across time, a murine model was assessed at three time points (5, 8, and 11 months). Transgenic mice with the G37R mutation of human SOD1 exhibited earliest signs of dysfunction at 8 months in terms of a pathological hindpaw clasping reflex, as well as slowed movement time on a suspended bar, anomalies in footprint patterns, weaker grip strength, raised somatosensory thresholds, and deficits in passive avoidance learning, yielding a margin of 3-4 months before death to test experimental therapies.

Anomalies in social behaviors and exploratory activities in an APPswe/PS1 mouse model of Alzheimer's disease.

Alzheimer's disease is characterized by deficits in social communication, associated with generalized apathy or agitation, as well as social memory. To assess social behaviors in 6-month-old male APPswe/PS1 bigenics relative to non-transgenic controls, the 3-chamber test was used, together with open-field and elevated plus-maze tests of exploration. APPswe/PS1 mice were less willing to engage in social interaction than wild-type, avoiding an unfamiliar stimulus mouse, probably not due to generalized apathy because in both tests of exploratory activity the mutants were hyperactive. This study reveals reduced "sociability" combined with hyperactivity in an APPswe/PS1 mouse model of Alzheimer dementia.

Subchronic memantine administration on spatial learning, exploratory activity, and nest-building in an APP/PS1 mouse model of Alzheimer's disease.

Glutamate neurotoxicity has been proposed to be involved in Alzheimer pathogenesis, with clinical data supporting successful treatment with the NMDA receptor antagonist memantine. In the present study, the effects of subchronic memantine administration were assessed on spatial and non-spatial learning as well as exploratory activity and nest-building in APP/PS1 mutant mice. Memantine (10 mg/kg, i.p.) was better than placebo during the reversal phase of left-right discrimination, though equivalent to saline for Morris water maze and passive avoidance learning. The drug had no effect on non-learned behaviors in elevated plus-maze exploration and nest-building. These results support a specific action of the NMDA receptor antagonist on behavioral flexibility in mutant mice with amyloid pathology.