Effects of P301L-TAU on post-translational modifications of microtubules in human iPSC-derived cortical neurons and TAU transgenic mice

TAU is a microtubule-associated protein that promotes microtubule assembly and stability in the axon.TAU is missorted and aggregated in an array of diseases known as tauopathies.Microtubules are essential for neuronal function and regulated via a complex set of post-translational modifications,changes of which affect microtubule stability and dynamics,microtubule interaction with other proteins and cellular structures,and mediate recruitment of microtubule-severing enzymes.As impairment of microtubule dynamics causes neuronal dysfunction,we hypothesize cognitive impairment in human disease to be impacted by impairment of microtubule dynamics.We therefore aimed to study the effects of a disease-causing mutation of TAU(P301L)on the levels and localization of microtubule post-translational modifications indicative of microtubule stability and dynamics,to assess whether P301L-TAU causes stability-changing modifications to microtubules.To investigate TAU localization,phosphorylation,and effects on tubulin post-translational modifications,we expressed wild-type or P301L-TAU in human MAPT-KO induced pluripotent stem cell-derived neurons(i Neurons)and studied TAU in neurons in the hippocampus of mice transgenic for human P301L-TAU(p R5 mice).Human neurons expressing the longest TAU isoform(2N4R)with the P301L mutation showed increased TAU phosphorylation at the AT8,but not the p-Ser-262 epitope,and increased polyglutamylation and acetylation of microtubules compared with endogenous TAU-expressing neurons.P301L-TAU showed pronounced somatodendritic presence,but also successful axonal enrichment and a similar axodendritic distribution comparable to exogenously expressed 2N4R-wildtype-TAU.P301L-TAU-expressing hippocampal neurons in transgenic mice showed prominent missorting and tauopathy-typical AT8-phosphorylation of TAU and increased polyglutamylation,but reduced acetylation,of microtubules compared with non-transgenic littermates.In sum,P301L-TAU results in changes in microtubule PTMs,suggestive of impairment of microtubule stability.This is accompanied by missorting and aggregation of TAU in mice but not in i Neurons.Microtubule PTMs/impairment may be of key importance in tauopathies.

Investigating Müller glia reprogramming in mice: a retrospective of the last decade, and a look to the future

Müller glia,as prominent glial cells within the retina,plays a significant role in maintaining retinal homeostasis in both healthy and diseased states.In lower vertebrates like zebrafish,these cells assume responsibility for spontaneous retinal regeneration,wherein endogenous Müller glia undergo proliferation,transform into Müller glia-derived progenitor cells,and subsequently regenerate the entire retina with restored functionality.Conversely,Müller glia in the mouse and human retina exhibit limited neural reprogramming.Müller glia reprogramming is thus a promising strategy for treating neurodegenerative ocular disorders.Müller glia reprogramming in mice has been accomplished with remarkable success,through various technologies.Advancements in molecular,genetic,epigenetic,morphological,and physiological evaluations have made it easier to document and investigate the Müller glia programming process in mice.Nevertheless,there remain issues that hinder improving reprogramming efficiency and maturity.Thus,understanding the reprogramming mechanism is crucial toward exploring factors that will improve Müller glia reprogramming efficiency,and for developing novel Müller glia reprogramming strategies.This review describes recent progress in relatively successful Müller glia reprogramming strategies.It also provides a basis for developing new Müller glia reprogramming strategies in mice,including epigenetic remodeling,metabolic modulation,immune regulation,chemical small-molecules regulation,extracellular matrix remodeling,and cell-cell fusion,to achieve Müller glia reprogramming in mice.

AAV mediated carboxyl terminus of Hsp70 interacting protein overexpression mitigates the cognitive and pathological phenotypes of APP/PS1 mice

The E3 ubiquitin ligase,carboxyl terminus of heat shock protein 70(Hsp70)interacting protein(CHIP),also functions as a co-chaperone and plays a crucial role in the protein quality control system.In this study,we aimed to investigate the neuroprotective effect of overexpressed CHIP on Alzheimer’s disease.We used an adeno-associated virus vector that can cross the blood-brain barrier to mediate CHIP overexpression in APP/PS1 mouse brain.CHIP overexpression significantly ameliorated the performance of APP/PS1 mice in the Morris water maze and nest building tests,reduced amyloid-βplaques,and decreased the expression of both amyloid-βand phosphorylated tau.CHIP also alleviated the concentration of microglia and astrocytes around plaques.In APP/PS1 mice of a younger age,CHIP overexpression promoted an increase in ADAM10 expression and inhibitedβ-site APP cleaving enzyme 1,insulin degrading enzyme,and neprilysin expression.Levels of HSP70 and HSP40,which have functional relevance to CHIP,were also increased.Single nuclei transcriptome sequencing in the hippocampus of CHIP overexpressed mice showed that the lysosomal pathway and oligodendrocyte-related biological processes were up-regulated,which may also reflect a potential mechanism for the neuroprotective effect of CHIP.Our research shows that CHIP effectively reduces the behavior and pathological manifestations of APP/PS1 mice.Indeed,overexpression of CHIP could be a beneficial approach for the treatment of Alzheimer’s disease.

Treadmill exercise in combination with acousto-optic and olfactory stimulation improves cognitive function in APP/PS1 mice through the brain-derived neurotrophic factor-and Cygb-associated signaling pathways

A reduction in adult neurogenesis is associated with behavioral abnormalities in patients with Alzheimer's disease.Consequently,enhancing adult neurogenesis represents a promising therapeutic approach for mitigating disease symptoms and progression.Nonetheless,nonpharmacological interventions aimed at inducing adult neurogenesis are currently limited.Although individual non-pharmacological interventions,such as aerobic exercise,acousto-optic stimulation,and olfactory stimulation,have shown limited capacity to improve neurogenesis and cognitive function in patients with Alzheimer's disease,the therapeutic effect of a strategy that combines these interventions has not been fully explored.In this study,we observed an age-dependent decrease in adult neurogenesis and a concurrent increase in amyloid-beta accumulation in the hippocampus of amyloid precursor protein/presenilin 1 mice aged 2-8 months.Amyloid deposition became evident at 4 months,while neurogenesis declined by 6 months,further deteriorating as the disease progressed.However,following a 4-week multifactor stimulation protocol,which encompassed treadmill running(46 min/d,10 m/min,6 days per week),40 Hz acousto-optic stimulation(1 hour/day,6 days/week),and olfactory stimulation(1 hour/day,6 days/week),we found a significant increase in the number of newborn cells(5'-bromo-2'-deoxyuridine-positive cells),immature neurons(doublecortin-positive cells),newborn immature neurons(5'-bromo-2'-deoxyuridine-positive/doublecortin-positive cells),and newborn astrocytes(5'-bromo-2'-deoxyuridine-positive/glial fibrillary acidic protein-positive cells).Additionally,the amyloid-beta load in the hippocampus decreased.These findings suggest that multifactor stimulation can enhance adult hippocampal neurogenesis and mitigate amyloid-beta neuropathology in amyloid precursor protein/presenilin 1 mice.Furthermore,cognitive abilities were improved,and depressive symptoms were alleviated in amyloid precursor protein/presenilin 1 mice following multifactor stimulation,as evidenced by Morris water maze,novel object recognition,forced swimming test,and tail suspension test results.Notably,the efficacy of multifactor stimulation in consolidating immature neurons persisted for at least 2weeks after treatment cessation.At the molecular level,multifactor stimulation upregulated the expression of neuron-related proteins(NeuN,doublecortin,postsynaptic density protein-95,and synaptophysin),anti-apoptosis-related proteins(Bcl-2 and PARP),and an autophagyassociated protein(LC3B),while decreasing the expression of apoptosis-related proteins(BAX and caspase-9),in the hippocampus of amyloid precursor protein/presenilin 1 mice.These observations might be attributable to both the brain-derived neurotrophic factor-mediated signaling pathway and antioxidant pathways.Furthermore,serum metabolomics analysis indicated that multifactor stimulation regulated differentially expressed metabolites associated with cell apoptosis,oxidative damage,and cognition.Collectively,these findings suggest that multifactor stimulation is a novel non-invasive approach for the prevention and treatment of Alzheimer's disease.

Gamma-glutamyl transferase 5 overexpression in cerebrovascular endothelial cells improves brain pathology,cognition,and behavior in APP/PS1 mice

In patients with Alzheimer’s disease,gamma-glutamyl transferase 5(GGT5)expression has been observed to be downregulated in cerebrovascular endothelial cells.However,the functional role of GGT5 in the development of Alzheimer’s disease remains unclear.This study aimed to explore the effect of GGT5 on cognitive function and brain pathology in an APP/PS1 mouse model of Alzheimer’s disease,as well as the underlying mechanism.We observed a significant reduction in GGT5 expression in two in vitro models of Alzheimer’s disease(Aβ_(1-42)-treated hCMEC/D3 and bEnd.3 cells),as well as in the APP/PS1 mouse model.Additionally,injection of APP/PS1 mice with an adeno-associated virus encoding GGT5 enhanced hippocampal synaptic plasticity and mitigated cognitive deficits.Interestingly,increasing GGT5 expression in cerebrovascular endothelial cells reduced levels of both soluble and insoluble amyloid-βin the brains of APP/PS1 mice.This effect may be attributable to inhibition of the expression ofβ-site APP cleaving enzyme 1,which is mediated by nuclear factor-kappa B.Our findings demonstrate that GGT5 expression in cerebrovascular endothelial cells is inversely associated with Alzheimer’s disease pathogenesis,and that GGT5 upregulation mitigates cognitive deficits in APP/PS1 mice.These findings suggest that GGT5 expression in cerebrovascular endothelial cells is a potential therapeutic target and biomarker for Alzheimer’s disease.

Serum interleukin-1 receptor antagonist is an early indicator of colitis onset in Gαi2-deficient mice

AIM： To study the serum concentration of IL-1β, IL-1 receptor antagonist （IL-1Ra） and IL-18 in Gαi2-deficient mice at the age of 6 （healthy）, 12 （pre-colitic） and 24 wk （coUtic） and in healthy control mice. METHODS： At the time of killing, serum samples were collected and IL-1β, IL-1Ra and IL-18 levels were measured using enzyme-linked immunosorbent assays. RESULTS： Serum concentration of IL-1Ra was significantly increased in pre-colitic （median： 524 ng/L; P= 0.02） and colitic （450 ng/L; P= 0.01）, but not in healthy （196 ng/L） Gαi2-deficient mice as compared with controls （217 ng/L）. Serum concentrations of IL-1β did not differ between Gαi2-deficient mice and their controls, irrespective of age, IL-18 was significantly increased in colitic, but not in pre-colitic mice compared with controls （510 ng/L vs 190 ng/L; P= 0.05）. CONCLUSION： The increased serum concentrations of IL-18 and IL-1Ra in established diseases are suggested as markers of ongoing colitis. Interestingly, the significantly increased serum concentration of IL-1Ra in pre-colitic mice is found to be an early marker of disease progression.

Assessment of the Effect of Environmental Factors on the Antagonism of <i>Bacillus amyloliquefaciens</i>and <i>Trichoderma harzianum</i>to <i>Colletotrichum acutatum</i>

The effect of temperature (18°C - 30°C), water activity (0.85 - 1) and pH (4 - 9) was studied by dual culture technique on the antagonism of Bacillus amyloliquefaciens and Trichoderma harzianum to Colletotrichum acutatum, responsible of strawberry (Fragaria x ananassa (Weston) Duchesne ex Rozier) anthracnose. The antagonistic bacteria’s strains behave significantly and differently according to the parameters studied. These results reveal useful information about the applicability of their biocontrol in agricultural culture with the change of environmental factors.

Studies on Rhizospheric Mycoflora of Tea (Camellia sinensis): In vitro Antagonism with Dominant Bacteria

Species of Trichoderma, Penicillium, Aspergillus and Mucor were found to dominate the rhizosphere of tea bushes in different regions of the Indian Himalayas. The Himachal Himalayas were dominated by Monilia humiscola (45.08%) and Mucor hiemalis (33.11%). In the Uttaranchal Himalayas, Aspergillus clavatum dominated the rhizosphere (54.01%), followed by A. flavous (20.07%). Although the fungal isolates require a mesophillic temperature (15 ℃ to 35 ℃), the tea rhizospheric isolates tolerate a wide range of temperatures (5 ℃ to 45 ℃). When tested for pH tolerance, these fungi showed a wide range (2.0 to 13.0) of pH tolerance under in vitro conditions. Seasonal variations also influenced the colony forming units (cfu) of the rhizospheric fungi, with the maximum forming in November and March, and the minimum in December^February. During this period, the populations of the fungi were found inversely proportional to the bacterial populations. In addition, the dominant bacteria in the tea rhizosphere, i.e., Bacillus subtilis and B. mycoides, showed antagonistic activity against fungal isolates by inhibiting the growth and causing structural abnormalities in mycelium. Tab 5, Ref

Identification and Antagonism Study of a Novel Chitinase-producing Bacterium Burkholderia Sp.C3 against Phytopathogenic Fungi

Through a modified agar well diffusion assay,antagonism of a novel chitinase-producing strain C3 against the phy- topathogenic fungi including Phoma wasabiae Yokogi,Cochlibolus Heterostrophus,Exserohilum Turcicum,Curuvularia Lunata (Walk)Boed,Thantephorus cucumris,Fusarium graminearum was tested.The data showed that the crude extracts of strain C3 had stable antifungal activity in the range of pH 5.0 to pH 8.0.The active components were heat labile and sensitive to proteinase K.A series of experiments supported that the compound responsible for inhibitory aetivity appeared to be chitinase.The 16s rDNA analysis indicated that C3 was subject to genus Burkholderia.Phenotypic characterization of C3 was also consisted with the result of molecular identification.

Identification of A Strain HN-1 against Banana Wilt Disease and Determination of Its Antagonism

[Objective] The paper was to identify strain HN-1 against banana wilt disease and to determine its antagonism. [Method] The strain HN-1 was ob- tained from the soil in fields heavily infected by Fusarium oxysporum f. sp. cubense （FOC）. Antagonism of the strain against F. oxysporum was tested via dual-cul- ture and inhibition test on spore germination. [Result] HN-1 effectively inhibited mycelial growth and spore germination of F. oxysporum, Strain HN-1 was identi- fied as BrevibaciUus brevis according to its characteristics in morphology, physiology and biochemistry and its 16S rDNA sequence. The strain showed high inhibition effect on 15 species of fungal pathogens in the dual-culture trials with fungal pathogens. [ Conclusion] The study provides theoretical basis for application of strain HN-1 in agricultural fields.

Growth and Antagonism of <i>Trichoderma</i>spp. and Conifer Pathogen <i>Heterobasidion annosum</i>s.l. <i>in Vitro</i>at Different Temperatures

Variations in the radial growth rate of 24 isolates belonging to ten species of Trichoderma, three isolates of conifer pathogen Heterobasidion annosum s.s. and four isolates of H. parviporum were evaluated by incubation on a solid malt extract medium at a temperature of 4℃, 15℃ and 21℃. Trichoderma antagonism against Heterobasidion was investigated in dual culture in vitro. The slowest rate of growth was referable to all seven strains of Heterobasidion spp. All Heterobasidion spp. strains were overgrown by 63% of Trichoderma spp. strains after two weeks at 21℃ and by 33% of strains at 15℃. 21% of Trichoderma strains did not grow and only four strains belonging to T. koningii, T. viride and T. viridescens demonstrated the ability to completely overgrow Heterobasidion spp. after two weeks incubation at 4℃. According to the antagonistic efficiency, Trichoderma strains were divided into five groups with an Euclidean distance of 25. The groups contained isolates from different species. It was suggested that selected psychrotrophic fast growing T. viride, T. koningii and T. viridescens strains could be examined in different substrate conditions as suitable antagonist agents for the control of H. annosum and H. parviporum.

Effect of Synergism and Antagonism between Metals on Toxicity in Soils

Synergism and antagonism of cadmium (Cd), copper (Cu) and selenium (Se) to biological toxicities in red soil, yellow brown soil and black soil were evaluated by MICROTOX method. The relation between forms of the tested metals in soil and the synergism or antagonism between them was also studied.Results showed that owing to the difference of soil chemical properties, toxicity of these metals in soils was different. In red soil with acid reaction and low in cation exchange capacity, antagonism occurred significantly between metals when they coexisted at high concentrations, while synergism occurred only under low concentrations. It is indicated that in red soil, toxicity of metals affected by synergism or antagonism depends on concentration of the metals present. For yellow brown soil and black soil with larger cation exchange capacity and lower exchangeable aluminium (A1), no toxicity of metals was observed even if metals were added to soil in high concentrations. Synergism and antagonism between Cd, Cu and Se were controlled by the forms of metals present. The amount of water-soluble metals was the most important factor in determining synergism and antagonism.In this paper, comparisons of synergism and antagonism between metals in soils and in water solutions were made. There occurred the synergism of metal toxicity in water solutions when the concentration of coexisting metals was high. This is just opposite to the case in soils.

Potential therapeutic role of spermine via Rac1 in osteoporosis:Insights from zebrafish and mice

Osteoporosis(OP)is a prevalent metabolic bone disease.While drug therapy is essential to prevent bone loss in osteoporotic patients,current treatments are limited by side effects and high costs,necessitating the development of more effective and safer targeted therapies.Utilizing a zebrafish(Danio rerio)larval model of osteoporosis,we explored the influence of the metabolite spermine on bone homeostasis.Results showed that spermine exhibited dual activity in osteoporotic zebrafish larvae by increasing bone formation and decreasing bone resorption.Spermine not only demonstrated excellent biosafety but also mitigated prednisolone-induced embryonic neurotoxicity and cardiotoxicity.Notably,spermine showcased protective attributes in the nervous systems of both zebrafish embryos and larvae.At the molecular level,Rac1 was identified as playing a pivotal role in mediating the antiosteoporotic effects of spermine,with P53 potentially acting downstream of Rac1.These findings were confirmed using mouse(Mus musculus)models,in which spermine not only ameliorated osteoporosis but also promoted bone formation and mineralization under healthy conditions,suggesting strong potential as a bonestrengthening agent.This study underscores the beneficial role of spermine in osteoporotic bone homeostasis and skeletal system development,highlighting pivotal molecular mediators.Given their efficacy and safety,human endogenous metabolites like spermine are promising candidates for new anti-osteoporotic drug development and daily bone-fortifying agents.

Effects of Rosa roxburghii&edible fungus fermentation broth on immune response and gut microbiota in immunosuppressed mice

With the rise of probiotics fermentation in food industry,fermented foods have attracted worldwide attention.In this study,protective effects of Rosa roxburghii&edible fungus fermentation broth(REFB)on immune function and gut health in Cyclophosphamide induced immunosuppressed mice were investigated.Results showed that REFB could improve the immune organ index,and promote the proliferation and differentiation of splenic T lymphocytes.In addition,it attenuated intestinal mucosal damage and improved intestinal cellular immunity.REFB administration also up-regulated the expression of IL-4,INF-γ,TNF-α,T-bet and GATA-3 mRNA in small intestine.Furthermore,administration of REFB modulated gut microbiota composition and increased the relative abundance of beneficial genus,such as Bacteroides.It also increased the production of fecal short-chain fatty acids.These indicate that REFB has the potential to improve immunity,alleviate intestinal injury and regulate gut microbiota in immunosuppressed mice.

Antagonism of local isolates of Trichoderma spp. on citrus root rot disease by Fusarium solani in the mekong delta of vietnam

The local isolates of Trichoderma spp. and Fusarium solani were colected from citrus orchards in the Mekong delta of Vietnam and isolated on PDA, PDB and TSM medium for antagonism and Koch’s postulate testing. The results showed that the high chitinolytic enzymes content of Trichoderma isolates can antagonise with Fusarium solani isolates by preventing the germination of Fusarium macroconidia in in-vitro condition. There are five promising isolates of Trichoderma spp. having high antagonism with Fusarium solani. These Trichoderma isolates also grew well in rice straws, maize stems, weeds and water hyacinth biowaste materials. These results supply the promising trend for biological control of root rot disease on citrus orchards of the Mekong delta.

WORLD MILITARY SITUATION 2016: ANTAGONISM INTENSIFIES BUT CRISES REMAIN CONTROLLABLE

The world military situation in 2016was complex,military strategies of major countries undergoing profound readjustment,contest in emerging fields increasing and a new round of populism cropping up worldwide which resulted in'Black swan events'happening one after another,major-country contest and geopolitical conflicts piling up and local

The Thromboxane Receptor Antagonist S18886 influence the stability of atherosclerosis in ApoE Null Mice

Objectives To investigate whether thromboxane receptor antagonist S18886 inhibits infiltrating macrophages to vessel wall and influence the morphology of atherosclerotic plaque; The effective of S18886 compared to clopidegrol on the development of atherosclerosis, accumulation of lipid- filled macropha-ges in apoE null mice. Methods All mice were done cuffed common carotid artery and fed a Western-type atherogenic diet for 6 weeks from the day of surgery, at same time the therapy group mice were gavaged S18886 5 mg/Kg/day and clopidegrol respec- tively, the same volume water were gavaged as the placebo group. Results profound inhibition of lesion area growth after cuff of the right common carotid artery in mice with 5 mg/kg of S18886, markdely reduce intima to media ratio and intima to total wall area compare with clopidegrol or blank group; Macrophage infiltration into sites of arterial plaque was also markedly attenuated by ICAM-1 deficiency in the S18886 group, whereas inside the arterial wall plaque of placebo apoE null mice α-smooth muscle actin markedly attenuated. Treatment with 25 mg/kg/day clopidegrol reduced the level of ICAM-1 stai ning, both S18886 and clopidegrol didn't influence the α-smooth muscle actin inside plaque. Conclusions It was considered that the novel anti-thrombotic drug significant reduce macrophage infiltration in the sites of arterial plaque by ICAM-1 deficiency, S18886 not only reduce the size, but also stabilized the plaque.

Application of transgenic mice to the molecular pathogenesis of cataract

One of the most prevalent disorders that cause blindness worldwide is cataract,and its essence is the visual disorder caused by the opacity of the lens.The significant degree of variation in cataracts and the fact that a variety of factors can impact a patient’s lens transparency make it especially crucial to investigate the pathogenesis of cataracts at the molecular level.It has been found that more than 60 genes are linked to the formation of cataracts,and the construction of a transgenic mouse model of cataract similar to the selection of human lens clouding due to a variety of causes has become an important means of studying the pathogenesis of cataract.Therefore,the research on the application of transgenic mice to the molecular pathogenesis of cataracts will be the main topic of this review of the literature.

Skeletal phenotypes and molecular mechanisms in aging mice

Aging is an inevitable physiological process,often accompanied by age-related bone loss and subsequent bone-related diseases that pose serious health risks.Research on skeletal diseases caused by aging in humans is challenging due to lengthy study durations,difficulties in sampling,regional variability,and substantial investment.Consequently,mice are preferred for such studies due to their similar motor system structure and function to humans,ease of handling and care,low cost,and short generation time.In this review,we present a comprehensive overview of the characteristics,limitations,applicability,bone phenotypes,and treatment methods in naturally aging mice and prematurely aging mouse models(including SAMP6,POLG mutant,LMNA,SIRT6,ZMPSTE24,TFAM,ERCC1,WERNER,and KL/KL-deficient mice).We also summarize the molecular mechanisms of these aging mouse models,including cellular DNA damage response,senescence-related secretory phenotype,telomere shortening,oxidative stress,bone marrow mesenchymal stem cell(BMSC)abnormalities,and mitochondrial dysfunction.Overall,this review aims to enhance our understanding of the pathogenesis of aging-related bone diseases.

Textile dyeing wastewater negatively influences the hematological profile and reproductive health of male Swiss albino mice

Objective:To determine the effects of textile dyeing industrial wastewater on the hematological parameters and reproductive health including histoarchitecture of male gonad(testes)of mice.Methods:Twenty-four Swiss albino mice at 4-weeks old were divided into four groups(n=6 per group).Mice of group 1 supplied with normal drinking water were served as the control group.Mice of group 2,3 and 4 were supplied normal drinking water mixed with textile dyeing wastewater at 5%,10% and 20% concentration,respectively.After completing 24 weeks of treatment,different hematological profile,weight of testes,gonadosomatic index(GSI),sperm concentration and morphology were measured.Moreover,histopathological changes in testes were examined.Results:Hematocrit value and hemoglobin concentrations were decreased in all groups of wastewater-treated mice compared to the control group.Likewise,weight of testes,GSI and sperm concentration were decreased significantly in wastewater-treated mice in comparison to the control group.The percentage of morphologically healthy epididymal sperm was significantly reduced in wastewater-treated mice.Histopathological examination revealed degenerative changes in seminiferous tubules,a smaller number of spermatogenic cells,elongation of seminiferous tubules and degenerative changes of seminiferous tubules in wastewater-treated mice.Conclusions:Textile dyeing wastewater has harmful effects on hematological profile and reproductive health of male mice.