CSF1R-mediated myeloid cell depletion shifts the ratio of motor cortical excitatory to inhibitory neurons in a multiple system atrophy model.

Motor cortical circuit functions depend on the coordinated fine-tuning of two functionally diverse neuronal populations: glutamatergic pyramidal neurons providing synaptic excitation and GABAergic interneurons adjusting the response of pyramidal neurons through synaptic inhibition. Microglia are brain resident macrophages which dynamically refine cortical circuits by monitoring perineuronal extracellular matrix and remodelling synapses. Previously, we showed that colony-stimulating factor 1 receptor (CSF1R)-mediated myeloid cell depletion extended the lifespan, but impaired motor functions of MBP29 mice, a mouse model for multiple system atrophy. In order to better understand the mechanisms underlying these motor deficits we characterized the microglial involvement in the cortical balance of GABAergic interneurons and glutamatergic pyramidal neurons in 4-months-old MBP29 mice following CSF1R inhibition for 12 weeks. Lack of myeloid cells resulted in a decreased number of COUP TF1 interacting protein 2-positive (CTIP2+) layer V pyramidal neurons, however in a proportional increase of calretinin-positive GABAergic interneurons in MBP29 mice. While myeloid cell depletion did not alter the expression of important presynaptic and postsynaptic proteins, the loss of cortical perineuronal net area was attenuated by CSF1R inhibition in MBP29 mice. These cortical changes may restrict synaptic plasticity and potentially modify parvalbumin-positive perisomatic input. Collectively, this study suggests, that the lack of myeloid cells shifts the neuronal balance toward an increased inhibitory connectivity in the motor cortex of MBP29 mice thereby potentially deteriorating motor functions.

Genetic deletion of Nogo/Rtn4 ameliorates behavioral and neuropathological outcomes in amyloid precursor protein transgenic mice.

The cognitive impairment in Alzheimer's disease (AD) is associated with synaptic loss, neuritic sprouting and altered neuroplasticity. Compensatory neuritic sprouting might be beneficial, while aberrant sprouting could contribute to the neurodegenerative process. Nogo (or Rtn4) is a major myelin-derived inhibitor of axonal sprouting in adult CNS. Recent evidence has implicated both the Reticulon family of proteins and a receptor for Nogo, NgR, in reducing amyloid-beta production, a key step in AD pathogenesis. To test the hypothesis that Nogo, as an inhibitor of axonal sprouting, modulates disease progression in a mouse model of AD, we introduced an APP transgene (a human APP minigene carrying the Swedish and Indiana mutations under the platelet-derived growth factor subunit B (PDGFB) promoter) into a Nogo null background and characterized the behavioral and neuropathological consequences. We found that deleting Nogo ameliorates learning and memory deficits of APP transgenic mice in the Morris water maze at an early/intermediate stage of the disease. Furthermore, deleting Nogo restored the expression levels of markers for synapto-dendritic complexity and axonal sprouting including synaptophysin, MAP2, GAP43 and neurofilament that are otherwise reduced in APP transgenic mice. Other aspects of disease progression including neuronal loss, astrogliosis, microgliosis and, importantly, Abeta levels and amyloid deposits were not significantly altered by Nogo deletion. These data support the hypothesis that Nogo-mediated inhibition of neuritic sprouting contributes to the disease progression in an APP transgenic model of AD in a way that is mechanistically distinct from what has been proposed for Rtn3 or NgR.

Amelioration of the cerebrovascular amyloidosis in a transgenic model of Alzheimer's disease with the neurotrophic compound cerebrolysin.

Increased production and reduced clearance of amyloid beta (Abeta) plays a central role in the pathogenesis of Alzheimer's disease (AD). We have recently shown that the neurotrophic peptide mixture Cerebrolysin (Cbl) has the ability of improving synaptic functioning and reducing amyloid deposition in a transgenic (tg) animal model of Alzheimer's disease (AD). Since in AD, potentially toxic Abeta aggregates accumulate not only around neurons but also in the blood vessels, then it is important to investigate whether bioactive compounds such as Cbl might have the capacity to ameliorate the age-related cerebral amyloid angiopathy (CAA) in tg models. To this end, tg mice expressing mutant human amyloid precursor protein (APP) under the Thy1 promoter were treated with Cbl or saline alone starting at 7 or 12 months of age for a total of three months. Neuropathological analysis with an antibody against Abeta showed that Cbl decreased amyloid deposition around the blood vessels in a time dependent manner. These effects were accompanied by a reduction in perivascular microgliosis and astrogliosis and increased expression of markers of vascular fitness such as CD31 and ZO-1. No lymphocytic infiltration was observed associated with Abeta in the vessels. Consistent with these findings, ultrastructural analysis showed that while in tg mice treated with saline alone there was an abundant accumulation of amyloid fibers in the vascular wall accompanied by thickening of the basal membrane and endothelial cell damage, in Cbl-treated mice there was considerable reduction in the subcellular alterations of endothelial and smooth muscle cells with preservation of basal membranes and intercellular junctions. Taken together, these results suggest that Cbl treatment might have beneficial effects in patients with cognitive impairment due to cerebrovascular amyloidosis by reducing Abeta accumulation and promoting the preservation of the cerebrovasculature.

An antiaggregation gene therapy strategy for Lewy body disease utilizing beta-synuclein lentivirus in a transgenic model.

Current experimental gene therapy approaches for Parkinson's disease (PD) and dementia with Lewy bodies (DLB) include the use of viral vectors expressing antiapoptosis genes, neurotrophic factors and dopaminergic system enzymes. However, since increasing evidence favors a role for alpha-synuclein accumulation in the pathogenesis of these disorders, an alternative therapy might require the transfer of genes that might block alpha-synuclein accumulation. beta-Synuclein, the nonamyloidogenic homologue of alpha-synuclein, has recently been identified as a potential candidate. Thus, in vivo transfer of genes encoding beta-synuclein might provide a novel approach to the development of experimental treatments for PD and DLB. To assess this possibility and to better understand the mechanisms involved, a lentiviral vector expressing human (h) beta-synuclein (lenti-beta-synuclein) was tested in a transgenic (tg) mouse model of halpha-synuclein aggregation. This study showed that unilateral intracerebral injection of lenti-beta-synuclein reduced the formation of halpha-synuclein inclusions and the accumulation of halpha-synuclein in synapses and ameliorated the neurodegenerative alterations in the tg mice. Both in vivo and in vitro coimmunoprecipitation and immunoblot experiments show that the mechanisms of beta-synuclein neuroprotection involve binding of this molecule to halpha-synuclein and Akt, resulting in the decreased aggregation and accumulation of halpha-synuclein in the synaptic membrane. Together, these data further support a role for beta-synuclein in regulating the conformational state of alpha-synuclein and suggest that this gene transfer approach might have potential for the development of alternative therapies for PD and DLB.

The neuroprotective effects of Cerebrolysin in a transgenic model of Alzheimer's disease are associated with improved behavioral performance.

Cerebrolysin is a peptide mixture with neurotrophic effects that might have the ability of both reducing amyloid burden and improving synaptic plasticity in Alzheimer's disease (AD). In order to determine if Cerebrolysin is capable of ameliorating the neurodegenerative and behavioral alterations associated with amyloid beta (A beta) production; transgenic (tg) mice expressing mutant human amyloid precursor protein (APP) under the Thy1 promoter were treated with Cerebrolysin or saline alone starting at 3 or 6 months of age for a total of three months. Animals were then tested behaviorally (at 6 and 9 months of age respectively) in the water maze and then analyzed neuropathologically for amyloid burden, synaptic density, astrogliosis and apoptosis. Performance analysis in the water maze showed that in the younger tg mice cohort, Cerebrolysin treatment significantly ameliorated the performance deficits. In the older cohort, there was a trend toward improved performance in the learning curve. Neuropathological examination showed that in both age/treatment groups, Cerebrolysin promoted synaptic regeneration, and reduced the proportion of neurons displaying DNA fragmentation by the (TdT)-mediated dUTP-biotin nick-end labeling (TUNEL) method. Moreover, Cerebrolysin treatment reduced A beta burden by 43% in the young group and by 27% in the older group. Taken together, these results suggest that Cerebrolysin treatment might have beneficial effects in patients with cognitive impairment by reducing A beta accumulation and promoting the preservation of synaptic terminals.

Effects of Cerebrolysin on amyloid-beta deposition in a transgenic model of Alzheimer's disease.

We investigated the potential mechanisms through which Cerebrolysin, a neuroprotective noothropic agent, might affect Alzheimer's disease pathology. Transgenic (tg) mice expressing mutant human (h) amyloid precursor protein 751 (APP751) cDNA under the Thy-1 promoter (mThy1-hAPP751) were treated for four weeks with this compound and analyzed by confocal microscopy to asses its effects on amyloid plaque formation and neurodegeneration. In this model, amyloid plaques in the brain are found much earlier (beginning at 3 months) than in other tg models. Quantitative computer-aided analysis with anti-amyloid-beta protein (A beta) antibodies, revealed that Cerebrolysin significantly reduced the amyloid burden in the frontal cortex of 5-month-old mice. Furthermore, Cerebrolysin treatment reduced the levels of A beta(1-42). This was accompanied by amelioration of the synaptic alterations in the frontal cortex of mThy1-hAPP751 tg mice. In conclusion, the present study supports the possibility that Cerebrolysin might have neuroprotective effects by decreasing the production of A beta(1-42) and reducing amyloid deposition.

Early formation of mature amyloid-beta protein deposits in a mutant APP transgenic model depends on levels of Abeta(1-42).

The main objective of the present study was to develop an alternative singly-transgenic (tg) hAPP model where amyloid deposition will occur at an earlier age. For this purpose, we generated lines of tg mice expressing hAPP751 cDNA containing the London (V717I) and Swedish (K670M/N671L) mutations under the regulatory control of the murine (m)Thy-1 gene (mThy1-hAPP751). In the brains of the highest (line 41) and intermediate (lines 16 and 11) expressers, high levels of hAPP expression were found in neurons in layers 4-5 of the neocortex, hippocampal CA1 and olfactory bulb. As early as 3-4 months of age, line 41 mice developed mature plaques in the frontal cortex, whereas at 5-7 months plaque formation extended to the hippocampus, thalamus and olfactory region. Ultrastructural and double-immunolabeling analysis confirmed that most plaques were mature and contained dystrophic neurites immunoreactive with antibodies against APP, synaptophysin, neurofilament and tau. In addition, a decrease in the number of synaptophysin-immunoreactive terminals was most prominent in the frontal cortex of mice from line 41. Mice from line 11 developed diffuse amyloid deposits at 11 months of age, whereas mice from line 16 did not show evidence of amyloid deposition. Analysis of Abeta by ELISA showed that levels of Abeta(1-40) were higher in mice that did not show any amyloid deposits (line 16), whereas Abeta(1-42) was the predominant species in tg animals from the lines showing plaque formation (lines 41 and 11). Taken together this study indicates that early onset plaque formation depends on levels of Abeta(1-42).

beta-Synuclein inhibits alpha-synuclein aggregation: a possible role as an anti-parkinsonian factor.

We characterized beta-synuclein, the non-amyloidogenic homolog of alpha-synuclein, as an inhibitor of aggregation of alpha-synuclein, a molecule implicated in Parkinson's disease. For this, doubly transgenic mice expressing human (h) alpha- and beta-synuclein were generated. In doubly transgenic mice, beta-synuclein ameliorated motor deficits, neurodegenerative alterations, and neuronal alpha-synuclein accumulation seen in halpha-synuclein transgenic mice. Similarly, cell lines transfected with beta-synuclein were resistant to alpha-synuclein accumulation. halpha-synuclein was coimmunoprecipitated with hbeta-synuclein in the brains of doubly transgenic mice and in the double-transfected cell lines. Our results raise the possibility that beta-synuclein might be a natural negative regulator of alpha-synuclein aggregation and that a similar class of endogenous factors might regulate the aggregation state of other molecules involved in neurodegeneration. Such an anti-amyloidogenic property of beta-synuclein might also provide a novel strategy for the treatment of neurodegenerative disorders.

Neurotrophic effects of Cerebrolysin in animal models of excitotoxicity.

Excitotoxicity might play an important role in neurodegenerative disorders such as Alzheimer's disease. In the mouse brain, kainic acid (KA) lesioning results in neurodegeneration patterns similar to those found in human disease. For this study, two sets of experiments were performed in order to determine if Cerebrolysin ameliorates the alterations associated with KA administration. In the first set of experiments, mice received intraperitoneal KA injections followed by Cerebrolysin administration, while in the second, mice were pretreated with Cerebrolysin for 4 weeks and then challenged with KA. Behavioral testing in the water maze and assessment of neuronal structure by laser scanning confocal microscopy showed a significant protection against KA lesions in mice pretreated with Cerebrolysin. In contrast, mice that received Cerebrolysin after KA injections did not show significant improvement. This study supports the contention that Cerebrolysin might have a neuroprotective effect in vivo against excitotoxicity.

Synaptotagmin and synaptic transmission alterations in apolipoprotein E-deficient mice.

1. Aged apoE-deficient mice and age-matched controls were tested for cognitive alterations in the Morris water maze. 2. Water maze results were correlated with in vivo electrophysiology and expression of the synaptic protein synaptotagmin (p65). 3. Compared to age-matched controls, apolipoprotein E-deficient mice displayed significant performance impairment accompanied by in vivo electrophysiological alterations in the dentate gyrus. 4. Apolipoprotein E-deficient mice also showed a significant increase in the synaptic protein, synaptotagmin, a synaptic calcium sensor involved in neurotransmitter release. 5. Cognitive impairments in these animals may be associated with decreased synaptic excitability in hippocampal neurons and the regulatory role of apolipoprotein E in synaptic function might be mediated by modulation of the expression of calcium sensor proteins.

Differential neurotrophic effects of apolipoprotein E in aged transgenic mice.

The present study seeked to determine whether the neurodegenerative and cognitive alterations in aged apolipoprotein E-deficient mice are differentially reversed by transgenic overexpression of human apolipoprotein-E3 vs. apolipoprotein-E4 in the background of deficient endogenous apolipoprotein E. These studies showed dendritic alterations in pyramidal neurons of apolipoprotein-E4 transgenic mice, similar to the ones observed in apolipoprotein E-deficient mice. However, these mice had a preserved density of synaptophysin-immunoreactive presynaptic terminals. In contrast, mice overexpressing apolipoprotein-E3 showed no synapto-dendritic alterations. Analysis of behavioral performance in the Morris water maze showed that while apolipoprotein E-deficient mice performed poorly, overexpression of apolipoprotein-E3 and, to a lower extent apolipoprotein-E4, resulted in an improved performance. This study supports the contention that, compared with apolipoprotein-E4, apolipoprotein-E3 might have a greater neurotrophic in vivo effect in aged mice.

Softening patterns of light cured glass ionomer cements.

OBJECTIVE: The aim of this study was to determine the effects of commonly used food simulating solutions and sodium hydroxide on the softening of light cured glass ionomer cements. METHODS: Four types of light cured glass ionomers (classified on the basis of the liquid component) as follows: (1) materials that combine a polymerizable monomer and polyalkenoic acid (PMPA); (2) use of a polymerizable polyalkenoic acid (PPA); (3) acid monomer (AM) in place of the polyalkenoic acid; and (4) replacement of polyalkenoic acid with polymerizable monomer (PMPR). A traditional glass ionomer and a microfil composite were used as controls. Disc-shaped specimens aged for a week at 37 degrees C and 100% relative humidity were stored in water, ethanol, heptane and 0.1 M sodium hydroxide for a period of 28 days. Barcol hardness measurements were made before immersion as well as at intervals of 24 h, 3 days, 7 days and 28 days after immersion. RESULTS: In general the softening effect was lowest on the resin composite control. Hardness could not be measured for the traditional glass ionomer after 24 h due to breakage and dissolution of samples. The different solutions had varying effects on the different classes of light cured glass ionomers. The change in hardness after 28 days ranged from an increase of +6.7% for PMPA material in heptane to a complete disintegration of PPA amd PMPR in NaOH at 60 degrees C. SIGNIFICANCE: The softening effect of food simulating solutions is dependent on the formulation of light cured glass ionomers. In clinical use, the role of softening in wear will consequently vary.

The effect of noble metals on the mechanical properties of dispersed phase dental amalgam.

The effects of noble metals added as part of a Ag-Cu dispersant on the compressive strength and creep of dental amalgams was determined. The Ag-Cu eutectic used in high copper dispersant alloy L(0), was altered by adding 15 per cent by weight (wt%) of noble metal in place of Ag. In L(1) the noble metal was Pd. In L(2), Au and Pd were combined in equal proportions. In L(3), the noble metal content was Pd and Pt in equal proportions. A low copper lathe-cut amalgam, Aristalloy was used as the matrix. Amalgams S(0) and S(1) had the same composition as L(0) and L(1) except that the the low copper alloy matrix was Spheralloy, a low copper spherical amalgam. The compressive strength and creep were measured according to American Dental Association specification No. 1 and the results analysed by ANOVA. The addition of noble metals to the dispersant significantly lowered the 1-h compressive strength of amalgams. Compared to the control amalgam L(0), the 24-h compressive strength was increased for L(1) and L(2) but lowered for L(3). The ADA creep values were significantly lowered by addition of all combinations of noble metals. Reported microstructural changes such as an increase in unreacted particles, and a slowed setting reaction may account for the findings.

Neurodegeneration and cognitive impairment in apoE-deficient mice is ameliorated by infusion of recombinant apoE.

Recent studies suggest that apolipoprotein E (apoE) might play a neurotrophic function in the central nervous system and that altered functioning of this molecule could result in neurodegeneration. The main objective of this study was to determine if neurodegenerative and cognitive alterations in apoE-deficient mice are reversible by infusion of recombinant apoE into the lateral ventricles. ApoE-deficient mice treated with either apoE3 or apoE4 showed a significant improvement in their learning capacity in the Morris water maze compared to saline-infused apoE-deficient mice. While this improved performance was associated with restoration of neuronal structure, the poor learning ability of apoE-deficient mice treated with saline correlated with the disrupted synapto-dendritic structure. This study supports the contention that apoE might play a neurotrophic effect in vivo and suggests that apoE might have a potential therapeutic role.

Marginal leakage of combinations of glass-ionomer and composite resin restorations.

This study compared the microleakage of four different combinations of glass-ionomer and composite resin restorations. Forty class V cavities were prepared in permanent posterior teeth. The gingival margin of the cavities were placed in cementum and the occlusal wall in enamel. The cavities were restored as either 1) light-cured glass-ionomer cement (LC-GIC); 2) chemically cured glass-ionomer cement (CC-GIC); 3) composite resin (with dentin bonding agent); or 4) light-cured glass-ionomer and composite resin (sandwich). Marginal microleakage was assessed by methylene blue dye penetration after thermocycling between 5 degrees C and 60 degrees C for 300 cycles. Microleakage of LC-GIC restorations and the dentin-bonded composite resin restorations were significantly lower than the CC-GIC and the LC-GIC sandwich technique restorations. There was no significant difference in the microleakage between the LC-GIC and the dentin-bonded composite resin restorations. The degree of microleakage of the sandwich restorations was significantly lower than the CC-GIC restorations. Light-cured glass-ionomer and dentin bonded composite resin restorations exhibited a lower degree of leakage than the glass-ionomer/composite sandwich and chemically cured glass-ionomer restorations at the gingival margins of class V restorations.

Softening patterns of post-cure heat-treated dental composites.

The softening of post-cure, heat-treated dental composites in various solutions was evaluated by Knoop hardness measurements. The samples were heated at 120 degrees C for 7 min immediately after curing and immersed in various solutions, water, ethanol, heptane and 0.1 N NaOH. Significant increases in hardness were observed for heated samples when compared to unheated samples. The various solutions softened both heated and unheated composites but heated samples were softened to a lesser degree. Plots of hardness of heated and unheated samples were nearly parallel, indicating that mechanisms such as penetration of the resin matrix, degradation of the silane coupling agent and fillers may be involved in the softening of both heated and unheated composites. Post-cure heat treatment improved the resistance to softening of dental composites.

[Pneumatosis cystoides intestinalis revisited]. / La pneumatose kystique intestinale reconsidérée.

Two cases of pneumatosis cystoides coli are reported. The authors present an anatomic and physiopathologic review of the entity and give the list of etiologies with the worse prognosis. The radiologic signs are described and their contribution to an accurate diagnosis is stressed, as it may avoid useless surgical interventions.

[Post-traumatic gas in children. Apropos of a case]. / Aéroportie post-traumatique de l'enfant. A propos d'un cas.

The authors report a case of post-traumatic gas in the portal vein in a child. They stress the rarity of this lesion in such circumstances and recall the radiological signs together with the differential diagnoses and the principal aetiologies.

Juxtapapillary duodenal diverticula and diverticula of the colon: is there a general "gastrointestinal diverticular disease"?

Both juxtapapillary duodenal diverticula and colon diverticula are acquired lesions, the pathogenesis of which is believed to involve the influence of high intraluminal pressure on loci minoris resistentiae in the gastrointestinal wall. We wanted to investigate whether juxtapapillary duodenal diverticula and colon diverticula occur independently, or whether they are part of a hypothetical general "gastrointestinal diverticular disease". 239 patients with juxtapapillary duodenal diverticula were identified in 2231 patients undergoing ERCP. Complete radiology data were available in 119/239 patients. Double contrast barium enema had been performed in 28/119 patients. In these patients, colon diverticula were present in 9/20 women and 1/8 men. The frequency of colon diverticula in these patients was compared with randomly chosen age- and sex-matched controls, for whom barium enema results were available. In these controls, 9/20 women and 1/8 men also had colon diverticula (n.s.). We conclude that after stratification for age and sex, the occurrence of colon diverticula is not higher in patients with juxtapapillary duodenal diverticula than in the general population. Juxtapapillary duodenal diverticula and colon diverticula thus occur independently. These data are not in favor of the existence of a general gastrointestinal diverticular disease.