BACKGROUND： Neural stem cell （NSC） survival is closely associated with cell apoptosis in ischemic-hypoxic regions following transplantation. Numerous studies have revealed that X-box binding protein 1 （XBP1） is a...BACKGROUND： Neural stem cell （NSC） survival is closely associated with cell apoptosis in ischemic-hypoxic regions following transplantation. Numerous studies have revealed that X-box binding protein 1 （XBP1） is a transcription factor during endoplasmic reticulum unfolded protein response and is essential for cell survival, differentiation, and anti-apoptotic effects. OBJECTIVE： To determine the effects of the XBP1 gene on NSC proliferation and apoptosis under hypoxic conditions following XBP1 gene transfection into rat embryonic hippocampal NSCs using recombinant adenovirus vector. DESIGN, TIME AND SETTING： In vitro experiments were performed at the Laboratory of Cell Biology of Jilin University and Laboratory of Proteomics, Department of Neurology, Jilin University China from September 2008 to November 2009. MATERIALS： Recombinant adenovirus package XBP1 gene and Ad-XBPl-enhanced green fluorescent protein plasmid （Guangzhou Easywin BioMed Technology, China）, rabbit anti-XBP1 and its target gene estrogen receptor degradation-enhancing a-mannosidase-like protein （EDEM） glucose-regulated protein 78 （GRP78）, anti-apoptotic molecule Bcl-2 and proapoptotic molecule Bax polyclonal antibody （Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA）, and COCI2 （Sigma, St. Louis, MO, USA） were used in the present study. METHODS： Hippocampi from embryonic, Sprague Dawley rats on gestational day 16 were harvested for NSC isolation and cloning, followed by immunofluorescence for Nestin and sub-culturing. The recombinant adenovirus Ad-XBPl-enhanced green fluorescent protein plasmid was transfected into rat embryonic hippocampal NSCs, and then CoCl2 was applied to induce hypoxia. MAIN OUTCOME MEASURES： Cell quantification and 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide colorimetric assay were utilized to detect proliferation in XBPl-transfected NSCs for 7 consecutive days. Western blot assay was utilized to quantify XBP1 GRP78, EDEM, Bcl-2, and Bax expression. Flow cytometry was used to measure apoptosis. RESULTS： NSC proliferation was significantly enhanced following XBP1 gene transfection （P 〈 0.05）. Under hypoxic conditions, GRP78, EDEM, and Bcl-2 levels increased, but Bax levels decreased. In addition, NSC apoptosis decreased following transfection （P 〈 0.05）. CONCLUSION： The XBP1 gene was successfully transfected into rat embryonic hippocampal NSCs using a recombinant adenovirus vector. NSC proliferation following transfection, as well as anti-apoptotic effects under hypoxia, was significantly increased.展开更多
AIM:To evaluate aspects of cognition impacted by individuals with and without normal tension glaucoma.METHODS:Fifty normal tension glaucoma(NTG)and 50 control patients≥50 y of age were recruited from the UCSF Departm...AIM:To evaluate aspects of cognition impacted by individuals with and without normal tension glaucoma.METHODS:Fifty normal tension glaucoma(NTG)and 50 control patients≥50 y of age were recruited from the UCSF Department of Ophthalmology.Demographic data and glaucoma parameters were extracted from electronic medical records for both groups.Tests of executive function[Executive Abilities:Measures and Instruments for Neurobehavioral Evaluation and Research(EXAMINER)]and learning and memory[California Verbal Learning Test–Second Edition(CVLT-II)]were administered to both NTG and controls.Race,handedness,best-corrected visual acuity,maximum intraocular pressure,optic nerve cup-todisc ratio,visual field and optic nerve optical coherence tomography parameters,and a measure of general health(Charlson Comorbidity Index)were compared between NTG and controls as well as within NTG subgroups.Multivariate linear regression was used to compare group performances on the EXAMINER battery and CVLT-II while controlling for age,sex,and years of education.RESULTS:NTG and controls were comparable with respect to age,sex,race,education,handedness,and the Charlson Comorbidity Index(P>0.05 for all).Performance on the EXAMINER composite score and the CVLT-II did not differ between NTG and controls(P>0.05 for both).CONCLUSION:This is the first prospective study in which the cognitive function of subject with NTG were evaluated using a comprehensive,computerized neurocognitive battery.Subjects with NTG do not perform worse than unaffected controls on tests of executive function,learning,and memor y.Results do not suppor t the hypothesis that individuals with NTG are at higher risk for cognitive dysfunction and/or dementia.展开更多
OBJECTIVE Dopamine receptors(DRs) are involved in the development and treatment of many neuropsychiatric disorders.Currently available dopaminergic drugs modulate both DRD2 and DRD3,leading to side effects and uncerta...OBJECTIVE Dopamine receptors(DRs) are involved in the development and treatment of many neuropsychiatric disorders.Currently available dopaminergic drugs modulate both DRD2 and DRD3,leading to side effects and uncertainty as to the roles each DR subtype plays physiologically.Our lab employed high throughput screening paradigms to discover highly selective modulators for the DRD3.METHODS The NIH Molecular Libraries Program 400,000 + small molecule library was screened using the Discove Rx Path Hunter?β-arrestin assay for compounds that activate the DRD3 without effects on the DRD2.Confirmation and counter-screens assessed selectivity and mechanisms of action.We identified 62 potential agonists,and chose the most promising to perform a structure-activity relationship(SAR) study to increase potency while maintaining selectivity.The lead compound identified through this process,ML417,was also characterized using bioluminescence resonance energy transfer(BRET)-based β-arrestin recruitment and G-protein activation assays as well as p-ERK assays.Potential neuroprotective properties of this compound were assessed using a SHSY5 Y neuronal cell model.RESULTS ML417 displays potent,DRD3-selective agonist activity in multiple functional assays.Binding and functional GPCR screens(>165 receptors) show ML417 has limited cross-reactivity with other GPCRs.ML417 also displays superior(compared to the reference compound pramipexole),dose-dependent protection against a decrease in neurite length induced by 10 μmol·L^(-1) of the neurotoxin,6-hydroxydopamine,in the SHSY5 Y cell model.CONCLUSION We have discovered and characterized ML417,a potent and highly selective DRD3 agonist.This compound will be useful as a research tool,and may prove useful as a therapeutic drug lead.展开更多
Described is a patient with concurrent discrete gliomas: a pleomorphic xanthoastrocytoma with anaplastic features and ananaplastic oligoastrocytoma. The distinct and morphologically dissimilar tumors demonstrated simi...Described is a patient with concurrent discrete gliomas: a pleomorphic xanthoastrocytoma with anaplastic features and ananaplastic oligoastrocytoma. The distinct and morphologically dissimilar tumors demonstrated similar genetic abnormalities by loss ofheterozygosity and comparative genome hybridization. Clonality and proteomic analyses highlighted an independent origin for thetwo tumors. Proteomic methods may prove useful in cases where the differential diagnosis and pathogenetic origin of tumors areuncertain, as well as more globally for its ability to provide insight into specific expression of proteins that may serve as uniquemarkers of tumorigenesis or as novel targets of therapy.展开更多
Cyclin-dependent kinase 5（Cdk5） is a member of the serine-threonine kinase family of cyclin-dependent kinases. Cdk5 is critical to normal mammalian nervous system development and plays important regulatory roles in ...Cyclin-dependent kinase 5（Cdk5） is a member of the serine-threonine kinase family of cyclin-dependent kinases. Cdk5 is critical to normal mammalian nervous system development and plays important regulatory roles in multiple cellular functions. Recent evidence indicates that Cdk5 is inappropriately activated in several neurodegenerative conditions, including Parkinson＇s disease（PD）. PD is a chronic neurodegenerative disorder characterized by the loss of dopamine neurons in the substantia nigra, decreased striatal dopamine levels, and consequent extrapyramidal motor dysfunction. During neurotoxicity, p35 is cleaved to form p25. Binding of p25 with Cdk5 leads deregulation of Cdk5 resulting in number of neurodegenerative pathologies. To date, strategies to specifically inhibit Cdk5 hyperactivity have not been successful without affecting normal Cdk5 activity. Here we show that inhibition of p25/Cdk5 hyperactivation through TFP5/TP5, truncated 24-aa peptide derived from the Cdk5 activator p35 rescues nigrostriatal dopaminergic neurodegeneration induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP/MPP＋） in a mouse model of PD. TP5 peptide treatment also blocked dopamine depletion in the striatum and improved gait dysfunction after MPTP administration. The neuroprotective effect of TFP5/TP5 peptide is also associated with marked reduction in neuroinflammation and apoptosis. Here we show inhibition of Cdk5/p25-hyperactivation by TFP5/TP5 peptide, which identifies Cdk5/p25 as a potential therapeutic target to reduce neurodegeneration in PD.展开更多
The goal of this work was to investigate how the MEG signal amplitude correlates with that of BOLD fMRI. To investigate the correlation between fMRI and macroscopic electrical activity, BOLD fMRI and MEG was performed...The goal of this work was to investigate how the MEG signal amplitude correlates with that of BOLD fMRI. To investigate the correlation between fMRI and macroscopic electrical activity, BOLD fMRI and MEG was performed on the same subjects (n=5). A visual flicker stimulus of varying temporal frequency was used to elicit neural responses in early visual areas. A strong similarity was observed in frequency tuning curves between both modalities. Although, averaged over subjects, the BOLD tuning curve was somewhat broader than MEG, both BOLD and MEG had maxima at a flicker frequency of 10 Hz. Also, we measured the first and second harmonic components as the stimuli frequency by MEG. In the low stimuli frequency (less than 6 Hz), the second harmonic has comparable amplitude with the first harmonic, which implies that neural frequency response is nonlinear and has more nonlinear components in low frequency than in high frequency.展开更多
Sleep is a widely expressed behavior across the animal kingdom. In addition to the numerous health benefits that are associated with sleep, it is believed that sleep plays a pivotal role in mental processes such as le...Sleep is a widely expressed behavior across the animal kingdom. In addition to the numerous health benefits that are associated with sleep, it is believed that sleep plays a pivotal role in mental processes such as learning and memory. Indeed, it has been demonstrated that learning and memory benefit from sleep, whereas sleep loss causes cognitive impairment(Rasch and Born, 2013). Changing the strength of synapses.展开更多
Recently there have been exciting research advances in neuroprotective therapies for ischemic stroke. In the past, the search for neu- roprotective agents has been fraught with failure at the clinical trials stage due...Recently there have been exciting research advances in neuroprotective therapies for ischemic stroke. In the past, the search for neu- roprotective agents has been fraught with failure at the clinical trials stage due to numerous factors, including subject heterogeneity and improper therapeutic windows （Tymianski, 2017）. Moreover, it is becoming clearer that the complex and evolving pathobiology of stroke requires multimodal therapeutic approaches capable of modulating the numerous axes that contribute to ischemia/reperfusion damage, rather than targeting a single axis （Bernstock et al., 2018a）. With the success of recent endovascular thrombectomy （EVT） trials, it has been suggested that clinical trials of EVT with adjunct neuroprotection can overcome past difficulties and maximize the effect size by using imaging to reduce patient heterogeneity （i. e., selecting those with large vessel occlusions, small ischemic cores, and good collateral circulation）, restoring perfusion using better EVT devices, and enrolling patients in the correct therapeutic window （i.e., when they still have salvageable brain tissue） （Tymianski, 2017）. Considering the opportunity that this represents for new, better clinical trials of neuroprotective agents, the search is on for high-potential compounds that may be investigated in these future studies.展开更多
Acetylcholine is an essential neurotransmitter found throughout the nervous system. Its action on postsynaptic receptors is regulated through hydrolysis by various carboxylesterases, especially cholinesterases (ChEs)....Acetylcholine is an essential neurotransmitter found throughout the nervous system. Its action on postsynaptic receptors is regulated through hydrolysis by various carboxylesterases, especially cholinesterases (ChEs). The acute toxicity of organophosphate (OP) compounds is directly linked to their action as inhibitors of ChE. One widely used assay for evaluating ChE activity is a spectrophotometric method developed by Ellman et al. When the enzyme source is from tissues or, in particular, blood, hemoglobin displays a spectrophotometric peak at the same wave-length used to analyze cholinergic activity. This creates a substantial background that interferes with the Ellman’s assay and must be overcome in order to accurately monitor cholinesterase activity. Herein, we directly compare blood processing methods: classical method (1.67 ± 0.30 U/mL) and HemogloBindTM treatment (1.51 ± 0.17 U/mL), and clearly demonstrate that pretreatment of blood samples with HemoglobindTM is both a sufficient and rapid sample preparation method for the assessment of ChE activity using the Ellman’s method.展开更多
Introduction Patients who had a mild ischaemic stroke who present with subtle or resolving symptoms sometimes go undiagnosed,are excluded from treatment and in some cases clinically worsen.Circulating immune cells are...Introduction Patients who had a mild ischaemic stroke who present with subtle or resolving symptoms sometimes go undiagnosed,are excluded from treatment and in some cases clinically worsen.Circulating immune cells are potential biomarkers that can assist with diagnosis in ischaemic stroke.Understanding the transcriptomic changes of each cell population caused by ischaemic stroke is critical because they work closely in a complicated relationship.In this study,we investigated peripheral blood mononuclear cells(PBMCs)transcriptomics of patients who had a stroke using a single-cell RNA sequencing to understand peripheral immune response after mild stroke based on the gene expression in an unbiased way.Methods Transcriptomes of PBMCsfrom 10 patients who had an acute ischaemic stroke within 24 hours after stroke onset were compared with 9 race-matched/age-matched/gender-matched controls.Individual PBMCs were prepared with ddSeqTM(Illumina-BioRad)and sequenced on the Illumina NovaSeq 6000 platform.Results Notable population changes were observed in patients who had a stroke,especially in NK cells and CD14+monocytes.The number of NK cells was increased,which was further confirmed by flow cytometry.Functional analysis implied that the activity of NK cells also is enhanced in patients who had a stroke.CD14+monocytes were clustered into two groups;dendritic cell-related CD14+monocytes and NK cell-related CD14+monocytes.We found CD14+monocyte subclusters were dramatically reduced in patients who had a stroke.Discussion This is the first study demonstrating the increased number of NK cells and new monocyte subclusters of mild ischaemic stroke based on the transcriptomic analysis.Our findings provide the dynamics of circulating immune response that could assist diagnosis and potential therapeutic development of mild ischaemic stroke.展开更多
Hypertonia is a neurological dysfunction associated with a number of central nervous system disorders, including cerebral palsy, Parkinson＇s disease, dystonia, and epilepsy. Genetic studies have identified a homozygo...Hypertonia is a neurological dysfunction associated with a number of central nervous system disorders, including cerebral palsy, Parkinson＇s disease, dystonia, and epilepsy. Genetic studies have identified a homozygous truncation mutation in Trakl that causes hypertonia in mice. Moreover, elevated Trakl protein expression is associated with several types of cancers and variants in Trakl are linked to childhood absence epilepsy in humans. Despite the importance of Trakl in health and disease, the mechanisms of Trakl action remain unclear and the pathogenic effects of Trakl mutation are unknown. Here we report that Trakl has a crucial function in regulation of mitochondrial fusion. Depletion of Trakl inhibits mitochondrial fusion, result- ing in mitochondrial fragmentation, whereas overex- pression of Trakl elongates and enlarges mitochondria. Our analyses revealed that Trakl interacts and colocal- izes with mitofusins on the outer mitochondrial mem- brane and functions with mitofusins to promote mitochondrial tethering and fusion. Furthermore, Trakl is required for stress-induced mitochondrial hyperfu- sion and pro-survival response. We found that hyper- tonia-associated mutation impairs Trakl mitochondrial localization and its ability to facilitate mitochondrial tethering and fusion. Our findings uncover a novel function of Trakl as a regulator of mitochondrial fusion and provide evidence linking dysregulated mitochon- drial dynamics to hypertonia pathogenesis.展开更多
Central nervous system(CNS)neurons typically fail to regenerate their axons after injury leading to neurological impairment.Axonal regeneration is a highly energy-demanding cellular program that requires local mitocho...Central nervous system(CNS)neurons typically fail to regenerate their axons after injury leading to neurological impairment.Axonal regeneration is a highly energy-demanding cellular program that requires local mitochondria to supply most energy within injured axons.Recent emerging lines of evidence have started to reveal that injury-triggered acute mitochondrial damage and local energy crisis contribute to the intrinsic energetic restriction that accounts for axon regeneration failure in the CNS.Characterizing and reprogramming bioenergetic signaling and mitochondrial maintenance after axon injury-ischemia is fundamental for developing therapeutic strategies that can restore local energy metabolism and thus facilitate axon regeneration.Therefore,establishing reliable and reproduc-ible neuronal model platforms is critical for assessing axonal energetic metabolism and regeneration capacity after injury-ischemia.In this focused methodology article,we discuss recent advances in applying cutting-edge microflu-idic chamber devices in combination with state-of-the-art live-neuron imaging tools to monitor axonal regeneration,mitochondrial transport,bioenergetic metabolism,and local protein synthesis in response to injury-ischemic stress in mature CNS neurons.展开更多
Tendon and ligament injuries are the most common musculoskeletal injuries,which not only impact the quality of life but result in a massive economic burden.Surgical interventions for tendon/ligament injuries utilize b...Tendon and ligament injuries are the most common musculoskeletal injuries,which not only impact the quality of life but result in a massive economic burden.Surgical interventions for tendon/ligament injuries utilize biological and/or engineered grafts to reconstruct damaged tissue,but these have limitations.Engineered matrices confer superior physicochemical properties over biological grafts but lack desirable bioactivity to promote tissue healing.While incorporating drugs can enhance bioactivity,large matrix surface areas and hydrophobicity can lead to uncontrolled burst release and/or incomplete release due to binding.To overcome these limitations,we evaluated the delivery of a peptide growth factor(exendin-4;Ex-4)using an enhanced nanofiber matrix in a tendon injury model.To overcome drug surface binding due to matrix hydrophobicity of poly(caprolactone)(PCL)-which would be expected to enhance cell-material interactions-we blended PCL and cellulose acetate(CA)and electrospun nanofiber matrices with fiber diameters ranging from 600 to 1000 nm.To avoid burst release and protect the drug,we encapsulated Ex-4 in the open lumen of halloysite nanotubes(HNTs),sealed the HNT tube endings with a polymer blend,and mixed Ex-4-loaded HNTs into the polymer mixture before electrospinning.This reduced burst release from~75%to~40%,but did not alter matrix morphology,fiber diameter,or tensile properties.We evaluated the bioactivity of the Ex-4 nanofiber formulation by culturing human mesenchymal stem cells(hMSCs)on matrix surfaces for 21 days and measuring tenogenic differentiation,compared with nanofiber matrices in basal media alone.Strikingly,we observed that Ex-4 nanofiber matrices accelerated the hMSC proliferation rate and elevated levels of sulfated glycosaminoglycan,tendon-related genes(Scx,Mkx,and Tnmd),and ECM-related genes(Col-Ⅰ,Col-Ⅲ,and Dcn),compared to control.We then assessed the safety and efficacy of Ex-4 nanofiber matrices in a full-thickness rat Achilles tendon defect with histology,marker expression,functional walking track analysis,and mechanical testing.Our analysis confirmed that Ex-4 nanofiber matrices enhanced tendon healing and reduced fibrocartilage formation versus nanofiber matrices alone.These findings implicate Ex-4 as a potentially valuable tool for tendon tissue engineering.展开更多
We present results of real-time and sensitive MR Thermometry (MRT) using a paramagnetic lanthanide complex thulium 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetramethyl-1,4,7,10-tetraa-cetate (Tm-DOTMA) to study radio f...We present results of real-time and sensitive MR Thermometry (MRT) using a paramagnetic lanthanide complex thulium 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetramethyl-1,4,7,10-tetraa-cetate (Tm-DOTMA) to study radio frequency (RF) heating induced by a copper wire and a titanium bone screw in an agarose gel phantom. The temperature dependent chemical shift coefficient (TDCSC) of the methyl resonance was found to be 0.7 ± 0.03 ppm/°;C in agarose gel. The methyl protons of Tm-DOTMA were imaged using 2D chemical shift imaging (CSI) and 3D phase mapping methods (PMM), approximately 7 sec long, and compared with conventional water proton resonance frequency (PRF) method. Two RF-induced heating approaches were tested: 1) using a prescan before the MRT;or 2) using the heating caused by the imaging pulse during continuous imaging. Both approaches allowed detection of temperature changes which are less than 1°;C and continuously mapping temperature changes around the copper wire. Using a heating pre-scan, the Tm-DOTMA 2D-CSI allowed better qualitative visualization of the temperature changes around the titanium screw compared with water phase shift thermometry. Numerical electromagnetic field simulations were also conducted for the evaluation of orientation dependency using the copper wire in 4.7 T (200 MHz). Thermometry approach using Tm-DOTMA can detect smaller temperature changes with decreased scanning time resulting in real-time and sensitive temperature mapping.展开更多
Background: Chronic, hypointense black holes (BHs) are recognized as a sign of permanent damage in patients with multiple sclerosis. Although the effects of interferon beta-1b in reducing the formation of new BHs are ...Background: Chronic, hypointense black holes (BHs) are recognized as a sign of permanent damage in patients with multiple sclerosis. Although the effects of interferon beta-1b in reducing the formation of new BHs are established, it is not clear whether the drug may reduce BH duration after these lesions are formed. Objective: To analyze the effects of interferon beta-1b in reducing the duration of T1 BHs in patients with multiple sclerosis. Design: Patients were clinically assessed and imaged monthly over a 36-month natural history phase and 36-month therapy phase. Numbers of contrast-enhanced lesions and newly formed BHs were counted on each scan. Each BH was counted until it was no longer seen. Setting: Outpatient service of the Neuroimmunology Branch at the National Institutes of Health, Bethesda, Md. Patients: Six patients with relapsing-remitting multiple sclerosis were included. One patient did not form any BHs during the therapy phase. Analyses were performed on the remaining 5 individuals. Interventions: Interferon beta-1b at the dosage of 8 million international units every other day. Main Outcome Measures: Number and duration (in months) of newly formed BHs. Res ults: Rate of BH accumulation decreased with treatment (P=.01), but Kaplan-Meier models revealed that the duration of BHs did not shorten (=2.47, P=.12). Conclusions: Interferon beta-1b reduces the frequency of new BH formation but does not appear to decrease their duration in time. Analyses with larger patient cohorts are needed to confirm these preliminary findings.展开更多
Background. Fabry disease is an under-diagnosed, treatable, X-linked, multisystem disorder. Objectives. To test the hypothesis that quality of life and sweating are decreased among pediatric patients with Fabry diseas...Background. Fabry disease is an under-diagnosed, treatable, X-linked, multisystem disorder. Objectives. To test the hypothesis that quality of life and sweating are decreased among pediatric patients with Fabry disease, compared with control subjects, and to provide quantitative natural history data and novel clinical end points for therapeutic trials. Design. Prospective, cross-sectional, observational study. Setting. Referral to the National Institutes of Health. Participants. Twenty-five male childrenwith Fabry disease (mean age: 12.3±3.5 years) and 21 age-matched control subjects. Main Outcome Measures. Quality of life (measured with the Child Health Questionnaire) and sweating (assessed with the quantitative sudomotor axon reflex test). Results. Quality of life scores for pediatric patients < 10 years of age with Fabry disease, compared with published normative values, were 55±17 vs 83±19 for bodily pain and 62±19 vs 80±13 for mental health. Bodily pain scores for patients ≥10 years of age were 54±22 vs 74±23. Sweat volume in the Fabry disease group was 0.41±0.46 μL/mm2, compared with 0.65±0.44 μL/mm2 in the control group. Renal function, urinary protein excretion, and cardiac function and structure were normal for the majority of patients. The 3 patients with residual α-galactosidase A activity ≥1.5%of normal values were free of cornea verticillata and had normal serum and urinary globotriaosylceramide levels. All other children had glycolipid levels comparable to those of adult patients with Fabry disease. Acroparesthesia and cardiac abnormalities were generally present before anhidrosis and proteinuria. Mapping of the missense mutations on the crystallographic structure of α-galactosidase A revealed that the mutations were partially surface-exposed and distal to the active site among individuals with residual enzyme activity. Mutations associated with left ventricular hypertrophy (defined as left ventricular mass index of > 51 g/m2) were localized near the catalytic site of the enzyme. Conclusions. Despite the absence of major organ dysfunction, Fabry disease demonstrates significant morbidity already in childhood. We have identified important, potentially correctable or preventable, outcome measures for future therapeutic trials. Prevention of complications involving major organs should be the goal for long-term specific therapy.展开更多
Purpose::The pathogenesis of gastrinomas is largely unknown,and there is a lack of reliable genetic determinants that are useful to distinguish malignant and benign forms of this tumor or predict the prognosis of pati...Purpose::The pathogenesis of gastrinomas is largely unknown,and there is a lack of reliable genetic determinants that are useful to distinguish malignant and benign forms of this tumor or predict the prognosis of patients with this disease.Loss of heterozygosity(LOH)on chromosome 3p is reported to occur in pancreatic neuroendocrine tumors(PNETs)as well as in non-PNETs and its presence is reported to correlate with tumor prognosis in non-endocrine tumors.However,little data are available from prospective studies on gastrinomas.Experimental design::We assessed occurrence of 3p LOH in 24 gastrinomas and correlated its presence with tumor biological behavior and other clinicopathological features of gastrinomas.Results::Either 3p LOH or microsatellite instability involving 3p occurred in 11 of 24 tumors(46%).Seven(29%)gastrinomas had 3p LOH.Of the 7 gastrinomas with 3p LOH,5(71%)had 3p12 LOH with the marker D3S2406,which was the shortest region of highest overlap(SRO).Chromosome 3p LOH was not associated with aggressive biological behavior of gastrinomas or with poor prognosis of patients with gastrinoma.Similarly,3p12 LOH(SRO)was not correlated with aggressive growth of tumors and/or liver metastases.Conclusion::Gastrinomas have a relative high frequency of 3p12 LOH suggesting this area may harbor putative tumor suppressor gene(s),which may play a role in the tumorigenesis,but not aggressiveness,of a subset of these tumors.展开更多
Parkinson’s disease(PD)diagnosis is based solely on clinical presentation[1].Therefore,diagnostic and prog-nostic biomarkers are needed.In recent years,seeding aggregation assays(SAA)have demonstrated the ability to ...Parkinson’s disease(PD)diagnosis is based solely on clinical presentation[1].Therefore,diagnostic and prog-nostic biomarkers are needed.In recent years,seeding aggregation assays(SAA)have demonstrated the ability to discriminate PD from controls,showing also potential as a predictive marker[2].Furthermore,the development of modern proteomic techniques such as the proxim-ity extension assay(PEA)has enabled high throughput,highly sensitive multiplexing studies to become more prevalent.In the past year,several such studies identi-fied DOPA decarboxylase(DDC)as an analyte of par-ticular interest,demonstrating consistent elevation in Lewy body diseases and promising diagnostic potential.展开更多
Neuronal synapses are functional nodes in neural circuits.Their organization and activity define an individual＇s level of intelligence,emotional state and mental health.Changes in the structure and efficacy of synaps...Neuronal synapses are functional nodes in neural circuits.Their organization and activity define an individual＇s level of intelligence,emotional state and mental health.Changes in the structure and efficacy of synapses are the biological basis of learning and memory.However,investigation of the molecular architecture of synapses has been impeded by the lack of efficient techniques with sufficient resolution.Recent developments in state-of-the-art nano-imaging techniques have opened up a new window for dissecting the molecular organization of neuronal synapses with unprecedented resolution.Here,we review recent technological advances in nano-imaging techniques as well as their applications to the study of synapses,emphasizing super-resolution light microscopy and 3-dimensional electron tomography.展开更多
There has been a growing appreciation for freezing of gait as a disabling symptom that causes a significant burden in Parkinson’s disease. Previous research has highlighted some of the key components that underlie th...There has been a growing appreciation for freezing of gait as a disabling symptom that causes a significant burden in Parkinson’s disease. Previous research has highlighted some of the key components that underlie the phenomenon, but these reductionist approaches have yet to lead to a paradigm shift resulting in the development of novel treatment strategies. Addressing this issue will require greater integration of multi-modal data with complex computational modeling, but there are a number of critical aspects that need to be considered before embarking on such an approach. This paper highlights where the field needs to address current gaps and shortcomings including the standardization of definitions and measurement, phenomenology and pathophysiology, as well as considering what available data exist and how future studies should be constructed to achieve the greatest potential to better understand and treat this devastating symptom.展开更多
文摘BACKGROUND： Neural stem cell （NSC） survival is closely associated with cell apoptosis in ischemic-hypoxic regions following transplantation. Numerous studies have revealed that X-box binding protein 1 （XBP1） is a transcription factor during endoplasmic reticulum unfolded protein response and is essential for cell survival, differentiation, and anti-apoptotic effects. OBJECTIVE： To determine the effects of the XBP1 gene on NSC proliferation and apoptosis under hypoxic conditions following XBP1 gene transfection into rat embryonic hippocampal NSCs using recombinant adenovirus vector. DESIGN, TIME AND SETTING： In vitro experiments were performed at the Laboratory of Cell Biology of Jilin University and Laboratory of Proteomics, Department of Neurology, Jilin University China from September 2008 to November 2009. MATERIALS： Recombinant adenovirus package XBP1 gene and Ad-XBPl-enhanced green fluorescent protein plasmid （Guangzhou Easywin BioMed Technology, China）, rabbit anti-XBP1 and its target gene estrogen receptor degradation-enhancing a-mannosidase-like protein （EDEM） glucose-regulated protein 78 （GRP78）, anti-apoptotic molecule Bcl-2 and proapoptotic molecule Bax polyclonal antibody （Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA）, and COCI2 （Sigma, St. Louis, MO, USA） were used in the present study. METHODS： Hippocampi from embryonic, Sprague Dawley rats on gestational day 16 were harvested for NSC isolation and cloning, followed by immunofluorescence for Nestin and sub-culturing. The recombinant adenovirus Ad-XBPl-enhanced green fluorescent protein plasmid was transfected into rat embryonic hippocampal NSCs, and then CoCl2 was applied to induce hypoxia. MAIN OUTCOME MEASURES： Cell quantification and 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide colorimetric assay were utilized to detect proliferation in XBPl-transfected NSCs for 7 consecutive days. Western blot assay was utilized to quantify XBP1 GRP78, EDEM, Bcl-2, and Bax expression. Flow cytometry was used to measure apoptosis. RESULTS： NSC proliferation was significantly enhanced following XBP1 gene transfection （P 〈 0.05）. Under hypoxic conditions, GRP78, EDEM, and Bcl-2 levels increased, but Bax levels decreased. In addition, NSC apoptosis decreased following transfection （P 〈 0.05）. CONCLUSION： The XBP1 gene was successfully transfected into rat embryonic hippocampal NSCs using a recombinant adenovirus vector. NSC proliferation following transfection, as well as anti-apoptotic effects under hypoxia, was significantly increased.
基金Supported by the Core Grant for Vision Research and the Research to Prevent Blindness Unrestricted Grant to the UCSF Department of Ophthalmology(No.NIH-NEI EY002162)。
文摘AIM:To evaluate aspects of cognition impacted by individuals with and without normal tension glaucoma.METHODS:Fifty normal tension glaucoma(NTG)and 50 control patients≥50 y of age were recruited from the UCSF Department of Ophthalmology.Demographic data and glaucoma parameters were extracted from electronic medical records for both groups.Tests of executive function[Executive Abilities:Measures and Instruments for Neurobehavioral Evaluation and Research(EXAMINER)]and learning and memory[California Verbal Learning Test–Second Edition(CVLT-II)]were administered to both NTG and controls.Race,handedness,best-corrected visual acuity,maximum intraocular pressure,optic nerve cup-todisc ratio,visual field and optic nerve optical coherence tomography parameters,and a measure of general health(Charlson Comorbidity Index)were compared between NTG and controls as well as within NTG subgroups.Multivariate linear regression was used to compare group performances on the EXAMINER battery and CVLT-II while controlling for age,sex,and years of education.RESULTS:NTG and controls were comparable with respect to age,sex,race,education,handedness,and the Charlson Comorbidity Index(P>0.05 for all).Performance on the EXAMINER composite score and the CVLT-II did not differ between NTG and controls(P>0.05 for both).CONCLUSION:This is the first prospective study in which the cognitive function of subject with NTG were evaluated using a comprehensive,computerized neurocognitive battery.Subjects with NTG do not perform worse than unaffected controls on tests of executive function,learning,and memor y.Results do not suppor t the hypothesis that individuals with NTG are at higher risk for cognitive dysfunction and/or dementia.
基金supported by National Institute of Neurological Disorders and Stroke Intramural Research Program
文摘OBJECTIVE Dopamine receptors(DRs) are involved in the development and treatment of many neuropsychiatric disorders.Currently available dopaminergic drugs modulate both DRD2 and DRD3,leading to side effects and uncertainty as to the roles each DR subtype plays physiologically.Our lab employed high throughput screening paradigms to discover highly selective modulators for the DRD3.METHODS The NIH Molecular Libraries Program 400,000 + small molecule library was screened using the Discove Rx Path Hunter?β-arrestin assay for compounds that activate the DRD3 without effects on the DRD2.Confirmation and counter-screens assessed selectivity and mechanisms of action.We identified 62 potential agonists,and chose the most promising to perform a structure-activity relationship(SAR) study to increase potency while maintaining selectivity.The lead compound identified through this process,ML417,was also characterized using bioluminescence resonance energy transfer(BRET)-based β-arrestin recruitment and G-protein activation assays as well as p-ERK assays.Potential neuroprotective properties of this compound were assessed using a SHSY5 Y neuronal cell model.RESULTS ML417 displays potent,DRD3-selective agonist activity in multiple functional assays.Binding and functional GPCR screens(>165 receptors) show ML417 has limited cross-reactivity with other GPCRs.ML417 also displays superior(compared to the reference compound pramipexole),dose-dependent protection against a decrease in neurite length induced by 10 μmol·L^(-1) of the neurotoxin,6-hydroxydopamine,in the SHSY5 Y cell model.CONCLUSION We have discovered and characterized ML417,a potent and highly selective DRD3 agonist.This compound will be useful as a research tool,and may prove useful as a therapeutic drug lead.
文摘Described is a patient with concurrent discrete gliomas: a pleomorphic xanthoastrocytoma with anaplastic features and ananaplastic oligoastrocytoma. The distinct and morphologically dissimilar tumors demonstrated similar genetic abnormalities by loss ofheterozygosity and comparative genome hybridization. Clonality and proteomic analyses highlighted an independent origin for thetwo tumors. Proteomic methods may prove useful in cases where the differential diagnosis and pathogenetic origin of tumors areuncertain, as well as more globally for its ability to provide insight into specific expression of proteins that may serve as uniquemarkers of tumorigenesis or as novel targets of therapy.
文摘Cyclin-dependent kinase 5（Cdk5） is a member of the serine-threonine kinase family of cyclin-dependent kinases. Cdk5 is critical to normal mammalian nervous system development and plays important regulatory roles in multiple cellular functions. Recent evidence indicates that Cdk5 is inappropriately activated in several neurodegenerative conditions, including Parkinson＇s disease（PD）. PD is a chronic neurodegenerative disorder characterized by the loss of dopamine neurons in the substantia nigra, decreased striatal dopamine levels, and consequent extrapyramidal motor dysfunction. During neurotoxicity, p35 is cleaved to form p25. Binding of p25 with Cdk5 leads deregulation of Cdk5 resulting in number of neurodegenerative pathologies. To date, strategies to specifically inhibit Cdk5 hyperactivity have not been successful without affecting normal Cdk5 activity. Here we show that inhibition of p25/Cdk5 hyperactivation through TFP5/TP5, truncated 24-aa peptide derived from the Cdk5 activator p35 rescues nigrostriatal dopaminergic neurodegeneration induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP/MPP＋） in a mouse model of PD. TP5 peptide treatment also blocked dopamine depletion in the striatum and improved gait dysfunction after MPTP administration. The neuroprotective effect of TFP5/TP5 peptide is also associated with marked reduction in neuroinflammation and apoptosis. Here we show inhibition of Cdk5/p25-hyperactivation by TFP5/TP5 peptide, which identifies Cdk5/p25 as a potential therapeutic target to reduce neurodegeneration in PD.
文摘The goal of this work was to investigate how the MEG signal amplitude correlates with that of BOLD fMRI. To investigate the correlation between fMRI and macroscopic electrical activity, BOLD fMRI and MEG was performed on the same subjects (n=5). A visual flicker stimulus of varying temporal frequency was used to elicit neural responses in early visual areas. A strong similarity was observed in frequency tuning curves between both modalities. Although, averaged over subjects, the BOLD tuning curve was somewhat broader than MEG, both BOLD and MEG had maxima at a flicker frequency of 10 Hz. Also, we measured the first and second harmonic components as the stimuli frequency by MEG. In the low stimuli frequency (less than 6 Hz), the second harmonic has comparable amplitude with the first harmonic, which implies that neural frequency response is nonlinear and has more nonlinear components in low frequency than in high frequency.
基金the NIH/NINDS Intramural Research Program (to WL)the Starting Grant of Excellent Talents from Xuzhou Medical University (to KW)。
文摘Sleep is a widely expressed behavior across the animal kingdom. In addition to the numerous health benefits that are associated with sleep, it is believed that sleep plays a pivotal role in mental processes such as learning and memory. Indeed, it has been demonstrated that learning and memory benefit from sleep, whereas sleep loss causes cognitive impairment(Rasch and Born, 2013). Changing the strength of synapses.
文摘Recently there have been exciting research advances in neuroprotective therapies for ischemic stroke. In the past, the search for neu- roprotective agents has been fraught with failure at the clinical trials stage due to numerous factors, including subject heterogeneity and improper therapeutic windows （Tymianski, 2017）. Moreover, it is becoming clearer that the complex and evolving pathobiology of stroke requires multimodal therapeutic approaches capable of modulating the numerous axes that contribute to ischemia/reperfusion damage, rather than targeting a single axis （Bernstock et al., 2018a）. With the success of recent endovascular thrombectomy （EVT） trials, it has been suggested that clinical trials of EVT with adjunct neuroprotection can overcome past difficulties and maximize the effect size by using imaging to reduce patient heterogeneity （i. e., selecting those with large vessel occlusions, small ischemic cores, and good collateral circulation）, restoring perfusion using better EVT devices, and enrolling patients in the correct therapeutic window （i.e., when they still have salvageable brain tissue） （Tymianski, 2017）. Considering the opportunity that this represents for new, better clinical trials of neuroprotective agents, the search is on for high-potential compounds that may be investigated in these future studies.
文摘Acetylcholine is an essential neurotransmitter found throughout the nervous system. Its action on postsynaptic receptors is regulated through hydrolysis by various carboxylesterases, especially cholinesterases (ChEs). The acute toxicity of organophosphate (OP) compounds is directly linked to their action as inhibitors of ChE. One widely used assay for evaluating ChE activity is a spectrophotometric method developed by Ellman et al. When the enzyme source is from tissues or, in particular, blood, hemoglobin displays a spectrophotometric peak at the same wave-length used to analyze cholinergic activity. This creates a substantial background that interferes with the Ellman’s assay and must be overcome in order to accurately monitor cholinesterase activity. Herein, we directly compare blood processing methods: classical method (1.67 ± 0.30 U/mL) and HemogloBindTM treatment (1.51 ± 0.17 U/mL), and clearly demonstrate that pretreatment of blood samples with HemoglobindTM is both a sufficient and rapid sample preparation method for the assessment of ChE activity using the Ellman’s method.
基金supported by the Intramural Research Program of the National Institutes of Health/National Institute of Neurological Disorders and Stroke and National Institute of Nursing Research(NIH ZIANS003043 and NIH ZIANR000015).
文摘Introduction Patients who had a mild ischaemic stroke who present with subtle or resolving symptoms sometimes go undiagnosed,are excluded from treatment and in some cases clinically worsen.Circulating immune cells are potential biomarkers that can assist with diagnosis in ischaemic stroke.Understanding the transcriptomic changes of each cell population caused by ischaemic stroke is critical because they work closely in a complicated relationship.In this study,we investigated peripheral blood mononuclear cells(PBMCs)transcriptomics of patients who had a stroke using a single-cell RNA sequencing to understand peripheral immune response after mild stroke based on the gene expression in an unbiased way.Methods Transcriptomes of PBMCsfrom 10 patients who had an acute ischaemic stroke within 24 hours after stroke onset were compared with 9 race-matched/age-matched/gender-matched controls.Individual PBMCs were prepared with ddSeqTM(Illumina-BioRad)and sequenced on the Illumina NovaSeq 6000 platform.Results Notable population changes were observed in patients who had a stroke,especially in NK cells and CD14+monocytes.The number of NK cells was increased,which was further confirmed by flow cytometry.Functional analysis implied that the activity of NK cells also is enhanced in patients who had a stroke.CD14+monocytes were clustered into two groups;dendritic cell-related CD14+monocytes and NK cell-related CD14+monocytes.We found CD14+monocyte subclusters were dramatically reduced in patients who had a stroke.Discussion This is the first study demonstrating the increased number of NK cells and new monocyte subclusters of mild ischaemic stroke based on the transcriptomic analysis.Our findings provide the dynamics of circulating immune response that could assist diagnosis and potential therapeutic development of mild ischaemic stroke.
文摘Hypertonia is a neurological dysfunction associated with a number of central nervous system disorders, including cerebral palsy, Parkinson＇s disease, dystonia, and epilepsy. Genetic studies have identified a homozygous truncation mutation in Trakl that causes hypertonia in mice. Moreover, elevated Trakl protein expression is associated with several types of cancers and variants in Trakl are linked to childhood absence epilepsy in humans. Despite the importance of Trakl in health and disease, the mechanisms of Trakl action remain unclear and the pathogenic effects of Trakl mutation are unknown. Here we report that Trakl has a crucial function in regulation of mitochondrial fusion. Depletion of Trakl inhibits mitochondrial fusion, result- ing in mitochondrial fragmentation, whereas overex- pression of Trakl elongates and enlarges mitochondria. Our analyses revealed that Trakl interacts and colocal- izes with mitofusins on the outer mitochondrial mem- brane and functions with mitofusins to promote mitochondrial tethering and fusion. Furthermore, Trakl is required for stress-induced mitochondrial hyperfu- sion and pro-survival response. We found that hyper- tonia-associated mutation impairs Trakl mitochondrial localization and its ability to facilitate mitochondrial tethering and fusion. Our findings uncover a novel function of Trakl as a regulator of mitochondrial fusion and provide evidence linking dysregulated mitochon- drial dynamics to hypertonia pathogenesis.
基金This work was supported by grants from the“Young Talent Support Plan”of Xi’an Jiaotong University(71211222010704)to N.Huangthe Intramural Research Program of NINDS,NIH(ZIA NS003029 and ZIA NS002946)to Z.-H.Sheng.
文摘Central nervous system(CNS)neurons typically fail to regenerate their axons after injury leading to neurological impairment.Axonal regeneration is a highly energy-demanding cellular program that requires local mitochondria to supply most energy within injured axons.Recent emerging lines of evidence have started to reveal that injury-triggered acute mitochondrial damage and local energy crisis contribute to the intrinsic energetic restriction that accounts for axon regeneration failure in the CNS.Characterizing and reprogramming bioenergetic signaling and mitochondrial maintenance after axon injury-ischemia is fundamental for developing therapeutic strategies that can restore local energy metabolism and thus facilitate axon regeneration.Therefore,establishing reliable and reproduc-ible neuronal model platforms is critical for assessing axonal energetic metabolism and regeneration capacity after injury-ischemia.In this focused methodology article,we discuss recent advances in applying cutting-edge microflu-idic chamber devices in combination with state-of-the-art live-neuron imaging tools to monitor axonal regeneration,mitochondrial transport,bioenergetic metabolism,and local protein synthesis in response to injury-ischemic stress in mature CNS neurons.
文摘Tendon and ligament injuries are the most common musculoskeletal injuries,which not only impact the quality of life but result in a massive economic burden.Surgical interventions for tendon/ligament injuries utilize biological and/or engineered grafts to reconstruct damaged tissue,but these have limitations.Engineered matrices confer superior physicochemical properties over biological grafts but lack desirable bioactivity to promote tissue healing.While incorporating drugs can enhance bioactivity,large matrix surface areas and hydrophobicity can lead to uncontrolled burst release and/or incomplete release due to binding.To overcome these limitations,we evaluated the delivery of a peptide growth factor(exendin-4;Ex-4)using an enhanced nanofiber matrix in a tendon injury model.To overcome drug surface binding due to matrix hydrophobicity of poly(caprolactone)(PCL)-which would be expected to enhance cell-material interactions-we blended PCL and cellulose acetate(CA)and electrospun nanofiber matrices with fiber diameters ranging from 600 to 1000 nm.To avoid burst release and protect the drug,we encapsulated Ex-4 in the open lumen of halloysite nanotubes(HNTs),sealed the HNT tube endings with a polymer blend,and mixed Ex-4-loaded HNTs into the polymer mixture before electrospinning.This reduced burst release from~75%to~40%,but did not alter matrix morphology,fiber diameter,or tensile properties.We evaluated the bioactivity of the Ex-4 nanofiber formulation by culturing human mesenchymal stem cells(hMSCs)on matrix surfaces for 21 days and measuring tenogenic differentiation,compared with nanofiber matrices in basal media alone.Strikingly,we observed that Ex-4 nanofiber matrices accelerated the hMSC proliferation rate and elevated levels of sulfated glycosaminoglycan,tendon-related genes(Scx,Mkx,and Tnmd),and ECM-related genes(Col-Ⅰ,Col-Ⅲ,and Dcn),compared to control.We then assessed the safety and efficacy of Ex-4 nanofiber matrices in a full-thickness rat Achilles tendon defect with histology,marker expression,functional walking track analysis,and mechanical testing.Our analysis confirmed that Ex-4 nanofiber matrices enhanced tendon healing and reduced fibrocartilage formation versus nanofiber matrices alone.These findings implicate Ex-4 as a potentially valuable tool for tendon tissue engineering.
文摘We present results of real-time and sensitive MR Thermometry (MRT) using a paramagnetic lanthanide complex thulium 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetramethyl-1,4,7,10-tetraa-cetate (Tm-DOTMA) to study radio frequency (RF) heating induced by a copper wire and a titanium bone screw in an agarose gel phantom. The temperature dependent chemical shift coefficient (TDCSC) of the methyl resonance was found to be 0.7 ± 0.03 ppm/°;C in agarose gel. The methyl protons of Tm-DOTMA were imaged using 2D chemical shift imaging (CSI) and 3D phase mapping methods (PMM), approximately 7 sec long, and compared with conventional water proton resonance frequency (PRF) method. Two RF-induced heating approaches were tested: 1) using a prescan before the MRT;or 2) using the heating caused by the imaging pulse during continuous imaging. Both approaches allowed detection of temperature changes which are less than 1°;C and continuously mapping temperature changes around the copper wire. Using a heating pre-scan, the Tm-DOTMA 2D-CSI allowed better qualitative visualization of the temperature changes around the titanium screw compared with water phase shift thermometry. Numerical electromagnetic field simulations were also conducted for the evaluation of orientation dependency using the copper wire in 4.7 T (200 MHz). Thermometry approach using Tm-DOTMA can detect smaller temperature changes with decreased scanning time resulting in real-time and sensitive temperature mapping.
文摘Background: Chronic, hypointense black holes (BHs) are recognized as a sign of permanent damage in patients with multiple sclerosis. Although the effects of interferon beta-1b in reducing the formation of new BHs are established, it is not clear whether the drug may reduce BH duration after these lesions are formed. Objective: To analyze the effects of interferon beta-1b in reducing the duration of T1 BHs in patients with multiple sclerosis. Design: Patients were clinically assessed and imaged monthly over a 36-month natural history phase and 36-month therapy phase. Numbers of contrast-enhanced lesions and newly formed BHs were counted on each scan. Each BH was counted until it was no longer seen. Setting: Outpatient service of the Neuroimmunology Branch at the National Institutes of Health, Bethesda, Md. Patients: Six patients with relapsing-remitting multiple sclerosis were included. One patient did not form any BHs during the therapy phase. Analyses were performed on the remaining 5 individuals. Interventions: Interferon beta-1b at the dosage of 8 million international units every other day. Main Outcome Measures: Number and duration (in months) of newly formed BHs. Res ults: Rate of BH accumulation decreased with treatment (P=.01), but Kaplan-Meier models revealed that the duration of BHs did not shorten (=2.47, P=.12). Conclusions: Interferon beta-1b reduces the frequency of new BH formation but does not appear to decrease their duration in time. Analyses with larger patient cohorts are needed to confirm these preliminary findings.
文摘Background. Fabry disease is an under-diagnosed, treatable, X-linked, multisystem disorder. Objectives. To test the hypothesis that quality of life and sweating are decreased among pediatric patients with Fabry disease, compared with control subjects, and to provide quantitative natural history data and novel clinical end points for therapeutic trials. Design. Prospective, cross-sectional, observational study. Setting. Referral to the National Institutes of Health. Participants. Twenty-five male childrenwith Fabry disease (mean age: 12.3±3.5 years) and 21 age-matched control subjects. Main Outcome Measures. Quality of life (measured with the Child Health Questionnaire) and sweating (assessed with the quantitative sudomotor axon reflex test). Results. Quality of life scores for pediatric patients < 10 years of age with Fabry disease, compared with published normative values, were 55±17 vs 83±19 for bodily pain and 62±19 vs 80±13 for mental health. Bodily pain scores for patients ≥10 years of age were 54±22 vs 74±23. Sweat volume in the Fabry disease group was 0.41±0.46 μL/mm2, compared with 0.65±0.44 μL/mm2 in the control group. Renal function, urinary protein excretion, and cardiac function and structure were normal for the majority of patients. The 3 patients with residual α-galactosidase A activity ≥1.5%of normal values were free of cornea verticillata and had normal serum and urinary globotriaosylceramide levels. All other children had glycolipid levels comparable to those of adult patients with Fabry disease. Acroparesthesia and cardiac abnormalities were generally present before anhidrosis and proteinuria. Mapping of the missense mutations on the crystallographic structure of α-galactosidase A revealed that the mutations were partially surface-exposed and distal to the active site among individuals with residual enzyme activity. Mutations associated with left ventricular hypertrophy (defined as left ventricular mass index of > 51 g/m2) were localized near the catalytic site of the enzyme. Conclusions. Despite the absence of major organ dysfunction, Fabry disease demonstrates significant morbidity already in childhood. We have identified important, potentially correctable or preventable, outcome measures for future therapeutic trials. Prevention of complications involving major organs should be the goal for long-term specific therapy.
文摘Purpose::The pathogenesis of gastrinomas is largely unknown,and there is a lack of reliable genetic determinants that are useful to distinguish malignant and benign forms of this tumor or predict the prognosis of patients with this disease.Loss of heterozygosity(LOH)on chromosome 3p is reported to occur in pancreatic neuroendocrine tumors(PNETs)as well as in non-PNETs and its presence is reported to correlate with tumor prognosis in non-endocrine tumors.However,little data are available from prospective studies on gastrinomas.Experimental design::We assessed occurrence of 3p LOH in 24 gastrinomas and correlated its presence with tumor biological behavior and other clinicopathological features of gastrinomas.Results::Either 3p LOH or microsatellite instability involving 3p occurred in 11 of 24 tumors(46%).Seven(29%)gastrinomas had 3p LOH.Of the 7 gastrinomas with 3p LOH,5(71%)had 3p12 LOH with the marker D3S2406,which was the shortest region of highest overlap(SRO).Chromosome 3p LOH was not associated with aggressive biological behavior of gastrinomas or with poor prognosis of patients with gastrinoma.Similarly,3p12 LOH(SRO)was not correlated with aggressive growth of tumors and/or liver metastases.Conclusion::Gastrinomas have a relative high frequency of 3p12 LOH suggesting this area may harbor putative tumor suppressor gene(s),which may play a role in the tumorigenesis,but not aggressiveness,of a subset of these tumors.
基金supported in part by the Intramural Research Program of the National Institute on Aging(NIA),the Center for Alzheimer’s and Related Dementias(CARD),within the Intramural Research Program of the National Institute on Aging and the National Institute of Neurological Disorders and Stroke(AG000546).
文摘Parkinson’s disease(PD)diagnosis is based solely on clinical presentation[1].Therefore,diagnostic and prog-nostic biomarkers are needed.In recent years,seeding aggregation assays(SAA)have demonstrated the ability to discriminate PD from controls,showing also potential as a predictive marker[2].Furthermore,the development of modern proteomic techniques such as the proxim-ity extension assay(PEA)has enabled high throughput,highly sensitive multiplexing studies to become more prevalent.In the past year,several such studies identi-fied DOPA decarboxylase(DDC)as an analyte of par-ticular interest,demonstrating consistent elevation in Lewy body diseases and promising diagnostic potential.
基金supported by grants from the National Natural Science Foundation of China (30725017 and 30928003)MOST (2009CB941300)
文摘Neuronal synapses are functional nodes in neural circuits.Their organization and activity define an individual＇s level of intelligence,emotional state and mental health.Changes in the structure and efficacy of synapses are the biological basis of learning and memory.However,investigation of the molecular architecture of synapses has been impeded by the lack of efficient techniques with sufficient resolution.Recent developments in state-of-the-art nano-imaging techniques have opened up a new window for dissecting the molecular organization of neuronal synapses with unprecedented resolution.Here,we review recent technological advances in nano-imaging techniques as well as their applications to the study of synapses,emphasizing super-resolution light microscopy and 3-dimensional electron tomography.
基金a National Health and Medical Research Council Leadership Fellowship(1195830)has received research funding from The Michael J.Fox Foundation and the Australian Research Council.S.A.F.was supported by The Sartain Lanier Family Foundation.He has been a consultant for Lundbeck,Sunovion,Biogen,Impel,Acorda,and CereSpir.He has received education and research grants from Medtronic,Boston Scientific,Sun Pharmaceuticals Advanced Research Company,Biohaven,Impax,Lilly,US World Meds,Sunovion Therapeutics,Neurocrine,Vaccinex,Voyager,Jazz Pharmaceuticals,CHDI Foundation,The Michael J.Fox Foundation,National Institutes of Health(NIH),and Parkinson’s Foundation.He receives royalties from Demos,Blackwell Futura,Springer for textbooks,and UpToDate.Other support was from Signant(Bracket Global LLC)and CNS Ratings LLC.N.G.serves as consultant to Sionara,NeuroDerm,Pharma2B,Denali,Neuron23,Sanofi-Genzyme,Biogen,and AbbVie.He receives royalties from Lysosomal Therapeutics(LTI)and payment for lectures at AbbVie,Sanofi-Genzyme,and Movement Disorder Society.He received research support from The Michael J.Fox Foundation,the National Parkinson Foundation,European Union,and Israel Science Foundation,as well as from Teva NNE program,Biogen,and Ionis.He receives support from the Sieratzki Family Foundation and the Aufzien Academic Center in Tel-Aviv University.A.N.received funding from the European Commission,Research Foundation Flanders,King Baudouin Foundation,The Michael J.Fox Foundation,Jacques and Gloria Gossweiler Foundation,and KU Leuven Internal Research Funds.M.H.is an inventor of patents held by the NIH for an immunotoxin for the treatment of focal movement disorders and the H-coil for magnetic stimulationin relation to the latter,he has received license-fee payments from the NIH(from Brainsway).He is on the Medical Advisory Boards of CALA Health and Brainsway(both unpaid positions).He is on the editorial board of approximately 15 journals and receives royalties and/or honoraria from publishing from Cambridge University Press,Oxford University Press,Springer,Wiley,Wolters Kluwer,and Elsevier.He has research grants from Medtronic,Inc.for a study of deep brain stimulation for dystonia and CALA Health for studies of a device to suppress tremor.
文摘There has been a growing appreciation for freezing of gait as a disabling symptom that causes a significant burden in Parkinson’s disease. Previous research has highlighted some of the key components that underlie the phenomenon, but these reductionist approaches have yet to lead to a paradigm shift resulting in the development of novel treatment strategies. Addressing this issue will require greater integration of multi-modal data with complex computational modeling, but there are a number of critical aspects that need to be considered before embarking on such an approach. This paper highlights where the field needs to address current gaps and shortcomings including the standardization of definitions and measurement, phenomenology and pathophysiology, as well as considering what available data exist and how future studies should be constructed to achieve the greatest potential to better understand and treat this devastating symptom.
