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The pathological phenotypes of human TDP-43 transgenic mouse models are independent of downregulation of mouse Tdp-43.  

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Tar DNA binding protein 43 (TDP-43) is the major component of pathological deposits in frontotemporal lobar degeneration with TDP-43 inclusions (FTLD-TDP) and in amyotrophic lateral sclerosis (ALS). It has been reported that TDP-43 transgenic mouse models expressing human TDP-43 wild-type or ALS-associated mutations recapitulate certain ALS and FTLD pathological phenotypes. Of note, expression of human TDP-43 (hTDP-43) reduces the levels of mouse Tdp-43 (mTdp-43). However, it remained unclear whether the mechanisms through which TDP-43 induces ALS or FTLD-like pathologies resulted from a reduction in mTdp-43, an increase in hTDP-43, or a combination of both. In elucidating the role of mTdp-43 and hTDP-43 in hTDP-43 transgenic mice, we observed that reduction of mTdp-43 in non-transgenic mice by intraventricular brain injection of AAV1-shTardbp leads to a dramatic increase in the levels of splicing variants of mouse sortilin 1 and translin. However, the levels of these two abnormal splicing variants are not increased in hTDP-43 transgenic mice despite significant downregulation of mTdp-43 in these mice. Moreover, further downregulation of mTdp-43 in hTDP-43 hemizygous mice, which are asymptomatic, to the levels equivalent to that of mTdp-43 in hTDP-43 homozygous mice does not induce the pathological phenotypes observed in the homozygous mice. Lastly, the number of dendritic spines and the RNA levels of TDP-43 RNA targets critical for synapse formation and function are significantly decreased in symptomatic homozygous mice. Together, our findings indicate that mTdp-43 downregulation does not lead to a loss of function mechanism or account for the pathological phenotypes observed in hTDP-43 homozygous mice because hTDP-43 compensates for the reduction, and associated functions of mTdp-43. Rather, expression of hTDP-43 beyond a certain threshold leads to abnormal metabolism of TDP-43 RNA targets critical for neuronal structure and function, which might be responsible for the ALS or FTLD-like pathologies observed in homozygous hTDP-43 transgenic mice. PMID:23922830

Xu, Ya-Fei; Prudencio, Mercedes; Hubbard, Jaime M; Tong, Jimei; Whitelaw, Ena C; Jansen-West, Karen; Stetler, Caroline; Cao, Xiangkun; Song, John; Zhang, Yong-Jie

2013-01-01

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Loss of murine TDP-43 disrupts motor function and plays an essential role in embryogenesis  

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Abnormal TDP-43 aggregation is a prominent feature in the neuropathology of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration. Mutations in TARDBP, the gene encoding TDP-43, cause some cases of ALS. The normal function of TDP-43 remains incompletely understood. To better understand TDP-43 biology, we generated mutant mice carrying a genetrap disruption of Tardbp. Mice homozygous for loss of TDP-43 are not viable. TDP-43 deficient embryos die about day 7.5 of embryonic ...

Kraemer, Brian C.; Schuck, Theresa; Wheeler, Jeanna M.; Robinson, Linda C.; Trojanowski, John Q.; Lee, Virginia M. Y.; Schellenberg, Gerard D.

2010-01-01

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Neurotoxic effects of TDP-43 overexpression in C. elegans  

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RNA-binding protein TDP-43 has been associated with multiple neurodegenerative diseases, including amyotrophic lateral sclerosis and frontotemporal lobar dementia. We have engineered pan-neuronal expression of human TDP-43 protein in Caenorhabditis elegans, with the goal of generating a convenient in vivo model of TDP-43 function and neurotoxicity. Transgenic worms with the neuronal expression of human TDP-43 exhibit an ‘uncoordinated’ phenotype and have abnormal motorneuron synapses. Cae...

Ash, Peter E. A.; Zhang, Yong-jie; Roberts, Christine M.; Saldi, Tassa; Hutter, Harald; Buratti, Emanuele; Petrucelli, Leonard; Link, Christopher D.

2010-01-01

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TDP-43 protein in plasma may index TDP-43 brain pathology in Alzheimer’s disease and frontotemporal lobar degeneration  

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Autopsy studies have shown that about 55% of patients with frontotemporal lobar degeneration (FTLD) and 25% of patients with Alzheimer’s disease (AD) harbour TDP-43 immunoreactive pathological changes in their brains. Using ELISA, we investigated whether we could detect the presence, or increased amounts, of TDP-43 in plasma of patients with FTLD and AD compared to normal control subjects. We detected elevated levels of TDP-43 protein in plasma of 46% patients with FTLD with clinical fronto...

Foulds, Penelope; Mcauley, Erica; Gibbons, Linda; Davidson, Yvonne; Pickering-brown, Stuart M.; Neary, David; Snowden, Julie S.; Allsop, David; Mann, David M. A.

2008-01-01

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Novel monoclonal antibodies to normal and pathologically altered human TDP-43 proteins.  

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The RNA/DNA-binding protein, TDP-43, is the key component of ubiquitinated inclusions characteristic of amyotrophic lateral sclerosis (ALS) and the majority of frontotemporal lobar degeneration (FTLD-TDP) referred to collectively as TDP-43 proteinopathies. To further elucidate mechanisms of pathological TDP-43 processing and identify TDP-43 epitopes that could be useful as potential biomarkers of TDP-43 proteinopathies, we developed a panel of novel monoclonal antibodies (MAbs) directed at regions extending across the length of TDP-43. Here, we confirm previous observations that there is no or minimal accumulation of TDP-43 N-terminal domains in neocortical inclusions in human TDP-43 proteinopathy tissues and we identify a subset of these MAbs that are specific for human versus mouse TDP-43. Notably, one of these MAbs recognized an epitope that preferentially detected pathological TDP-43 inclusions with negligible reactivity for normal nuclear TDP-43 resembling anti-phospho-TDP-43 specific antibodies that only bind pathological TDP-43. Hence, we infer that this new MAb recognizes a phosphorylation independent but disease-specific pathologic conformation in abnormal TDP-43. These data suggest that the novel MAbs reported here will be useful for patient-oriented research as well as for studies of animal and cell-based models of TDP-43 proteinopathies including ALS and FTLD-TDP. PMID:24690345

Kwong, Linda K; Irwin, David J; Walker, Adam K; Xu, Yan; Riddle, Dawn M; Trojanowski, John Q; Lee, Virginia M Y

2014-01-01

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Expression of mutant TDP-43 induces neuronal dysfunction in transgenic mice  

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Full Text Available Abstract Background Abnormal distribution, modification and aggregation of transactivation response DNA-binding protein 43 (TDP-43 are the hallmarks of multiple neurodegenerative diseases, especially frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U and amyotrophic lateral sclerosis (ALS. Researchers have identified 44 mutations in the TARDBP gene that encode TDP-43 as causative for cases of sporadic and familial ALS http://www.molgen.ua.ac.be/FTDMutations/. Certain mutant forms of TDP-43, such as M337V, are associated with increased low molecular weight (LMW fragments compared to wild-type (WT TDP-43 and cause neuronal apoptosis and developmental delay in chick embryos. Such findings support a direct link between altered TDP-43 function and neurodegeneration. Results To explore the pathogenic properties of the M337V mutation, we generated and characterized two mouse lines expressing human TDP-43 (hTDP-43M337V carrying this mutation. hTDP-43M337V was expressed primarily in the nuclei of neurons in the brain and spinal cord, and intranuclear and cytoplasmic phosphorylated TDP-43 aggregates were frequently detected. The levels of TDP-43 LMW products of ~25 kDa and ~35 kDa species were also increased in the transgenic mice. Moreover, overexpression of hTDP-43M337V dramatically down regulated the levels of mouse TDP-43 (mTDP-43 protein and RNA, indicating TDP-43 levels are tightly controlled in mammalian systems. TDP-43M337V mice displayed reactive gliosis, widespread ubiquitination, chromatolysis, gait abnormalities, and early lethality. Abnormal cytoplasmic mitochondrial aggregates and abnormal phosphorylated tau were also detected in the mice. Conclusion Our novel TDP-43M337V mouse model indicates that overexpression of hTDP-43M337V alone is toxic in vivo. Because overexpression of hTDP-43 in wild-type TDP-43 and TDP-43M337V mouse models produces similar phenotypes, the mechanisms causing pathogenesis in the mutant model remain unknown. However, our results suggest that overexpression of the hTDP-43M337V can cause neuronal dysfunction due to its effect on a number of cell organelles and proteins, such as mitochondria and TDP-43, that are critical for neuronal activity. The mutant model will serve as a valuable tool in the development of future studies designed to uncover pathways associated with TDP-43 neurotoxicity and the precise roles TDP-43 RNA targets play in neurodegeneration.

Dickson Dennis W

2011-10-01

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Spatial patterns of TDP-43 neuronal cytoplasmic inclusions (NCI) in fifteen cases of frontotemporal lobar degeneration with TDP-43 proteinopathy (FTLD-TDP)  

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Neuronal cytoplasmic inclusions (NCI) immunoreactive for transactive response DNA-binding protein (TDP-43) are the pathological hallmark of frontotemporal lobar degeneration with TDP-43 proteinopathy (FTLD-TDP). We studied the spatial patterns of the TDP-43 immunoreactive NCI in the frontal and temporal cortex of 15 cases of FTLD-TDP. The NCI were distributed parallel to the tissue boundary predominantly in regular clusters 50–400 ?m in diameter. In five cortical areas, the size of the clu...

Armstrong, Richard A.; Cairns, Nigel J.

2011-01-01

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RNP2 of RNA Recognition Motif 1 Plays a Central Role in the Aberrant Modification of TDP-43  

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Phosphorylated and truncated TAR DNA-binding protein-43 (TDP-43) is a major component of ubiquitinated cytoplasmic inclusions in neuronal and glial cells of two TDP-43 proteinopathies, amyotrophic lateral sclerosis and frontotemporal lobar degeneration. Modifications of TDP-43 are thus considered to play an important role in the pathogenesis of TDP-43 proteinopathies. However, both the initial cause of these abnormal modifications and the TDP-43 region responsible for its aggregation remain u...

Takagi, Shinnosuke; Iguchi, Yohei; Katsuno, Masahisa; Ishigaki, Shinsuke; Ikenaka, Kensuke; Fujioka, Yusuke; Honda, Daiyu; Niwa, Jun-ichi; Tanaka, Fumiaki; Watanabe, Hirohisa; Adachi, Hiroaki; Sobue, Gen

2013-01-01

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TDP-43 toxicity in yeast.  

Science.gov (United States)

The budding yeast Saccharomyces cerevisiae is an emerging tool for investigating the molecular pathways that underpin several human neurodegenerative disorders associated with protein misfolding. Amyotrophic lateral sclerosis (ALS) is a devastating adult onset neurodegenerative disease primarily affecting motor neurons. The protein TDP-43 has recently been demonstrated to play an important role in the disease, however, the mechanisms by which TDP-43 contributes to pathogenesis are unclear. To explore the mechanistic details that result in aberrant accumulation of TDP-43 and to discover potential strategies for therapeutic intervention, we employed a yeast TDP-43 proteinopathy model system. These studies allowed us to determine the regions of TDP-43 required for aggregation and toxicity and to define the effects of ALS-linked mutant forms of TDP-43. We have also been able to harness the power of yeast genetics to identify potent modifiers of TDP-43 toxicity using high-throughput yeast genetic screens. Here, we describe the methods and approaches that we have used in order to gain insight into TDP-43 biology and its role in disease. These approaches are readily adaptable to other neurodegenerative disease proteins. PMID:21115123

Armakola, Maria; Hart, Michael P; Gitler, Aaron D

2011-03-01

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Accumulation of TAR DNA Binding Protein-43 (TDP-43) in Mild Cognitive Impairment and Alzheimer Disease  

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TAR DNA binding protein-43 (TDP-43) plays a central role in the neuropathology of frontotemporal lobar degeneration (FTLD-TDP) and amyotrophic lateral sclerosis, but the relationship between TDP-43 abnormalities and Alzheimer disease (AD) remains unclear. To determine whether TDP-43 can serve as a neuropathological marker of AD, we performed biochemical characterization and quantification of TDP-43 in homogenates from parietal neocortex of subjects with a clinical diagnosis of no cognitive im...

Tremblay, Cyntia; St-amour, Isabelle; Schneider, Julie; Bennett, David A.; Calon, Fre?de?ric

2011-01-01

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Neuronal sensitivity to TDP-43 overexpression is dependent on timing of induction  

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Ubiquitin-immunoreactive neuronal inclusions composed of TAR DNA binding protein of 43 kDa (TDP-43) are a major pathological feature of frontotemporal lobar degeneration (FTLD-TDP). In vivo studies with TDP-43 knockout mice have suggested that TDP-43 plays a critical, although undefined role in development. In the current report, we generated transgenic mice that conditionally express wild-type human TDP-43 (hTDP-43) in the forebrain and established a paradigm to examine the sensitivity of n...

Cannon, Ashley; Yang, Baoli; Knight, Joshua; Farnham, Ian M.; Zhang, Yongjie; Wuertzer, Charles A.; D’alton, Simon; Lin, Wen-lang; Castanedes-casey, Monica; Rousseau, Linda; Scott, Brittany; Jurasic, Michael; Howard, John; Yu, Xin; Bailey, Rachel

2012-01-01

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TDP-43 Redistribution Is an Early Event in Sporadic Amyotrophic Lateral Sclerosis  

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Abstract Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder consisting of progressive loss of motor neurons. TDP-43 has been identified as a component of ubiquitin-immunoreactive inclusions of motor neurons in ALS. We focused on the diffuse cytoplasmic TDP-43 immunoreactivity in ALS neurons, and quantitatively assessed it in comparison with skein/round TDP-43 and ubiquitin immunostaining in motor neurons of 30 sporadic ALS cases. The percentage of spinal motor neurons with cy...

Giordana, Maria Teresa; Magistrello, Michela; Buccinna, Barbara; Piccinini, Marco; Grifoni, Silvia; Lupino, Elisa; Rinaudo, Maria Teresa; Vercellino, Marco

2010-01-01

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Autophagy induction enhances TDP43 turnover and survival in neuronal ALS models.  

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Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) have distinct clinical features but a common pathology-cytoplasmic inclusions rich in transactive response element DNA-binding protein of 43 kDa (TDP43). Rare TDP43 mutations cause ALS or FTD, but abnormal TDP43 levels and localization may cause disease even if TDP43 lacks a mutation. Here we show that individual neurons vary in their ability to clear TDP43 and are exquisitely sensitive to TDP43 levels. To measure TDP43 clearance, we developed and validated a single-cell optical method that overcomes the confounding effects of aggregation and toxicity and discovered that pathogenic mutations shorten TDP43 half-life. New compounds that stimulate autophagy improved TDP43 clearance and localization and enhanced survival in primary murine neurons and in human stem cell-derived neurons and astrocytes harboring mutant TDP43. These findings indicate that the levels and localization of TDP43 critically determine neurotoxicity and show that autophagy induction mitigates neurodegeneration by acting directly on TDP43 clearance. PMID:24974230

Barmada, Sami J; Serio, Andrea; Arjun, Arpana; Bilican, Bilada; Daub, Aaron; Ando, D Michael; Tsvetkov, Andrey; Pleiss, Michael; Li, Xingli; Peisach, Daniel; Shaw, Christopher; Chandran, Siddharthan; Finkbeiner, Steven

2014-08-01

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TDP-43 toxicity in yeast  

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The budding yeast Saccharomyces cerevisiae is an emerging tool for investigating the molecular pathways that underpin several human neurodegenerative disorders associated with protein misfolding. Amyotrophic lateral sclerosis (ALS) is a devastating adult onset neurodegenerative disease primarily affecting motor neurons. The protein TDP-43 has recently been demonstrated to play an important role in the disease, however the mechanisms by which TDP-43 contributes to pathogenesis are unclear. To ...

Armakola, Maria; Hart, Michael P.; Gitler, Aaron D.

2011-01-01

15

A Drosophila model for TDP-43 proteinopathy  

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Neuropathology involving TAR DNA binding protein-43 (TDP-43) has been identified in a wide spectrum of neurodegenerative diseases collectively named as TDP-43 proteinopathy, including amyotrophic lateral sclerosis (ALS) and frontotemporal lobar dementia (FTLD). To test whether increased expression of wide-type human TDP-43 (hTDP-43) may cause neurotoxicity in vivo, we generated transgenic flies expressing hTDP-43 in various neuronal subpopulations. Expression in the fly eyes of the full-lengt...

Li, Yan; Ray, Payal; Rao, Elizabeth J.; Shi, Chen; Guo, Weirui; Chen, Xiaoping; Woodruff, Elvin A.; Fushimi, Kazuo; Wu, Jane Y.

2010-01-01

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TDP-43 interaction with the intracellular domain of amyloid precursor protein induces p53-associated apoptosis.  

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TAR DNA-binding protein 43 (TDP-43), an essential pathological protein in both amyotrophic later sclerosis (ALS) and frontotemporal lobar degeneration (FTLD), is expressed abnormally in Alzheimer's disease (AD). However, whether and how TDP-43 contributes the pathogenesis of AD remains unknown. We have shown here a colocalization between TDP-43 and the intracellular domain of APP (AICD) in the nucleus. Coimmunoprecipitation analysis showed an interaction between TDP-43 and AICD. Overexpression of TDP-43 in COS7 cells enhanced the transactivation of AICD in an APP-Gal4 luciferase reporter system. Real-time PCR analysis showed that cotransfection of TDP-43 and AICD in HEK293 cells increased P53 mRNA levels compared to either TDP-43-transfected or AICD-transfected cells. Moreover, cotransfection of TDP-43 and AICD in either N2a or COS7 cells showed increased numbers of apoptotic cells compared to either TDP-43-transfected or AICD-transfected cells, indicating that TDP-43 enhances AICD-mediated apoptosis in N2a or COS7 cells. Thus, TDP-43 may play a role in AD pathology through interaction with AICD. PMID:24721672

Wang, Jing; Yan, Ke; Wu, Zhi-Qiang; Zheng, Chuan-Yi; Xu, Ru-Xiang; Chen, Li-Hua; Wen, Zhong-Min; Zhao, He-Qing; Ma, Quan-Hong

2014-05-21

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Altered distributions of Gemini of coiled bodies and mitochondria in motor neurons of TDP-43 transgenic mice  

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TAR DNA-binding protein-43 (TDP-43), a DNA/RNA-binding protein involved in RNA transcription and splicing, has been associated with the pathophysiology of neurodegenerative diseases, including ALS. However, the function of TDP-43 in motor neurons remains undefined. Here we use both gain- and loss-of-function approaches to determine roles of TDP-43 in motor neurons. Mice expressing human TDP-43 in neurons exhibited growth retardation and premature death that are characterized by abnormal intra...

Shan, Xiu; Chiang, Po-min; Price, Donald L.; Wong, Philip C.

2010-01-01

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TDP-43 functions and pathogenic mechanisms implicated in TDP-43 proteinopathies  

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Given the critical role for TDP-43 in diverse neurodegenerative diseases including amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD-TDP), there has been a recent surge in efforts to understand the normal functions of TDP-43 and the molecular basis of dysregulation that occurs in TDP-43 proteinopathies. Here, we highlight recent findings examining TDP-43 molecular functions with particular emphasis on stress-mediated regulation of TDP-43 localization, putative do...

Cohen, Todd J.; Lee, Virginia M. Y.; Trojanowski, John Q.

2011-01-01

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TDP-43 Variants of Frontotemporal Lobar Degeneration  

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It has been only 5 years since the identification of TDP-43 as the major protein component of the ubiquitinated inclusions in FTLD-U. At that time, there were approximately a dozen papers about TDP-43; today, a “TDP-43” search reveals almost 600 papers. It is now clear that the majority of FTLD cases containing tau- and alpha-synuclein-negative, ubiquitin-positive inclusions (FTLD-U) are FTLD-TDP. The spectrum of TDP-43 proteinopathies includes FTLD-TDP with or without ALS, with or withou...

Bigio, Eileen H.

2011-01-01

20

Divergent Phenotypes in Mutant TDP-43 Transgenic Mice Highlight Potential Confounds in TDP-43 Transgenic Modeling  

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The majority of cases of frontotemporal lobar degeneration and amyotrophic lateral sclerosis are pathologically defined by the cleavage, cytoplasmic redistribution and aggregation of TAR DNA binding protein of 43 kDa (TDP-43). To examine the contribution of these potentially toxic mechanisms in vivo, we generated transgenic mice expressing human TDP-43 containing the familial amyotrophic lateral sclerosis-linked M337V mutation and identified two lines that developed neurological phenotypes of differing severity and progression. The first developed a rapid cortical neurodegenerative phenotype in the early postnatal period, characterized by fragmentation of TDP-43 and loss of endogenous murine Tdp-43, but entirely lacking aggregates of ubiquitin or TDP-43. A second, low expressing line was aged to 25 months without a severe neurodegenerative phenotype, despite a 30% loss of mouse Tdp-43 and accumulation of lower molecular weight TDP-43 species. Furthermore, TDP-43 fragments generated during neurodegeneration were not C-terminal, but rather were derived from a central portion of human TDP-43. Thus we find that aggregation is not required for cell loss, loss of murine Tdp-43 is not necessarily sufficient in order to develop a severe neurodegenerative phenotype and lower molecular weight TDP-43 positive species in mouse models should not be inherently assumed to be representative of human disease. Our findings are significant for the interpretation of other transgenic studies of TDP-43 proteinopathy.

D'Alton, Simon; Altshuler, Marcelle; Cannon, Ashley; Dickson, Dennis W.; Petrucelli, Leonard; Lewis, Jada

2014-01-01

 
 
 
 
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ALS and FTLD: two faces of TDP-43 proteinopathy  

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Major discoveries have been made in the recent past in the genetics, biochemistry and neuropathology of frontotemporal lobar degeneration (FTLD). TAR DNA-binding protein 43 (TDP-43), encoded by the TARDBP gene, has been identified as the major pathological protein of FTLD with ubiquitin-immunoreactive (ub-ir) inclusions (FTLD-U) with or without amyotrophic lateral sclerosis (ALS) and sporadic ALS. Recently, mutations in the TARDBP gene in familial and sporadic ALS have been reported which dem...

Liscic, R. M.; Grinberg, L. T.; Zidar, J.; Gitcho, M. A.; Cairns, N. J.

2008-01-01

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TDP-43-induced Death Is Associated with Altered Regulation of BIM and Bcl-xL and Attenuated by Caspase-mediated TDP-43 Cleavage*  

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Abnormal aggregates of transactive response DNA-binding protein-43 (TDP-43) and its hyperphosphorylated and N-terminal truncated C-terminal fragments (CTFs) are deposited as major components of ubiquitinated inclusions in most cases of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with ubiquitinated inclusions (FTLD-U). The mechanism underlying the contribution of TDP-43 to the pathogenesis of these neurodegenerative diseases remains unknown. In this study, we foun...

Suzuki, Hiroaki; Lee, Kikyo; Matsuoka, Masaaki

2011-01-01

23

Astrocytic TDP-43 Pathology in Alexander Disease.  

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Alexander disease (AxD) is a rare neurodegenerative disorder characterized pathologically by the presence of eosinophilic inclusions known as Rosenthal fibers (RFs) within astrocytes, and is caused by dominant mutations in the coding region of the gene encoding glial fibrillary acidic protein (GFAP). GFAP is the major astrocytic intermediate filament, and in AxD patient brain tissue GFAP is a major component of RFs. TAR DNA binding protein of 43 kDa (TDP-43) is the major pathological protein in almost all cases of the neurodegenerative disease amyotrophic lateral sclerosis (ALS) and ?50% of frontotemporal lobar degeneration (FTLD), designated as FTLD-TDP. In ALS and FTLD-TDP, TDP-43 becomes insoluble, ubiquitinated, and pathologically phosphorylated and accumulates in cytoplasmic inclusions in both neurons and glia of affected brain and spinal cord regions. Previously, TDP-43 was detected in RFs of human pilocytic astrocytomas; however, involvement of TDP-43 in AxD has not been determined. Here we show that TDP-43 is present in RFs in AxD patient brains, and that insoluble phosphorylated full-length and high molecular weight TDP-43 accumulates in white matter of such brains. Phosphorylated TDP-43 also accumulates in the detergent-insoluble fraction from affected brain regions of Gfap(R236H/+) knock-in mice, which harbor a GFAP mutation homologous to one that causes AxD in humans, and TDP-43 colocalizes with astrocytic RF pathology in Gfap(R236H/+) mice and transgenic mice overexpressing human wild-type GFAP. These findings suggest common pathogenic mechanisms in ALS, FTLD, and AxD, and this is the first report of TDP-43 involvement in a neurological disorder primarily affecting astrocytes. PMID:24806671

Walker, Adam K; Daniels, Christine M Lapash; Goldman, James E; Trojanowski, John Q; Lee, Virginia M-Y; Messing, Albee

2014-05-01

24

Molecular Neuropathology of TDP-43 Proteinopathies  

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Full Text Available The identification of TDP-43 as the major component of the pathologic inclusions in most forms of sporadic and familial frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U and amyotrophic lateral sclerosis (ALS resolved a long-standing enigma concerning the nature of the ubiquitinated disease protein under these conditions. Anti-TDP-43 immunohistochemistry and the recent development of novel tools, such as phosphorylation-specific TDP-43 antibodies, have increased our knowledge about the spectrum of pathological changes associated with FTLD-U and ALS and moreover, facilitated the neuropathological routine diagnosis of these conditions. This review summarizes the recent advances in our understanding on the molecular neuropathology and pathobiology of TDP-43 in FTLD and ALS.

Manuela Neumann

2009-01-01

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TARDBP mutation analysis in TDP-43 proteinopathies and deciphering the toxicity of mutant TDP-43  

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The identification of TAR DNA-binding protein 43 (TDP-43) as the major disease protein in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with ubiquitin inclusions (FTLD-U) has defined a new class of neurodegenerative conditions: the TDP-43 proteinopathies. This breakthrough was quickly followed by mutation analysis of TARDBP, the gene encoding TDP-43. Herein, we provide a review of our previously published efforts that led to the identification of 3 TARDBP mutations...

Gendron, Tania F.; Rademakers, Rosa; Petrucelli, Leonard

2013-01-01

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Rodent Models of TDP-43 Proteinopathy: Investigating the Mechanisms of TDP-43-Mediated Neurodegeneration  

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Since the identification of phosphorylated and truncated transactive response DNA-binding protein 43 (TDP-43) as a primary component of ubiquitinated inclusions in amyotrophic lateral sclerosis and frontotemporal lobar degeneration with ubiquitin-positive inclusions, much effort has been directed towards ascertaining how TDP-43 contributes to the pathogenesis of disease. As with other protein misfolding disorders, TDP-43-mediated neuronal death is likely caused by both a toxic gain and loss o...

Gendron, Tania F.; Petrucelli, Leonard

2011-01-01

27

Targeting TDP-43 in neurodegenerative diseases.  

Science.gov (United States)

Introduction: TAR DNA-binding protein-43 (TDP-43) is a ubiquitously expressed RNA-binding protein belonging to the hnRNP family of nuclear proteins. In human disease, its aberrant aggregation in brains has been shown to play a causative role in several neurodegenerative diseases, especially ALS and FTLD. Areas covered: In this work, we have highlighted what could be the most promising avenues that could be exploited in a profitable manner to modulate TDP-43 pathology. These range from its protein-protein interactions, RNA-protein interactions and its aberrant aggregation process. Recently published articles on these subjects have been reviewed in the writing up of this manuscript. Expert opinion: Targeting aberrant TDP-43 aggregation in neurodegenerative diseases should be considered both a challenge and an opportunity. The challenge is represented by the central role played by TDP-43 in the general cellular and developmental processes of higher proteins. This characteristic makes it difficult to target this protein in a generalized manner. In addition, and mostly because of this reason, we still lack reliable disease model systems that can reproduce most, if not all, characteristics of the human disease. Nonetheless, recent research is finally starting to provide potential therapeutic targets based on new findings that regard TDP-43 biology and functions. PMID:24649927

Budini, Mauricio; Baralle, Francisco E; Buratti, Emanuele

2014-06-01

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Widespread RNA metabolism impairment in sporadic inclusion body myositis TDP43-proteinopathy?  

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TDP43 protein mislocalization is a hallmark of the neurodegenerative diseases amyotrophic lateral sclerosis and frontotemporal dementia, and mutations in the gene encoding TDP43 cause both disorders, further highlighting its role in disease pathogenesis. TDP43 is a heterogenous ribonucleoprotein, therefore suggesting that alterations in RNA metabolism play a role in these disorders, although direct evidence in patients is lacking. Sporadic inclusion body myositis (sIBM) is the most common acquired myopathy occurring in adults aged older than 50 years and abnormal cytoplasmic accumulations of TDP43 have been consistently described in sIBM myofibers. Here, we exploit high quality RNA from frozen sIBM muscle biopsies for transcriptomic studies on TDP43-proteinopathy patient tissue. Surprisingly, we found widespread sIBM-specific changes in the RNA metabolism pathways themselves. Consistent with this finding, we describe novel RNA binding proteins to mislocalize in the cytoplasm of sIBM myofibers and splicing changes in MAPT, a gene previously shown to play a role in sIBM. Our data indicate widespread alterations of RNA metabolism are a novel aspect of disease pathogenesis in sIBM. These findings also document an association, in TDP43-proteinopathy patients, between heterogenous ribonucleoprotein pathology and RNA metabolism alterations and carry importance for neurodegenerative diseases, such as amyotrophic lateral sclerosis and frontotemporal dementia.

Cortese, Andrea; Plagnol, Vincent; Brady, Stefen; Simone, Roberto; Lashley, Tammaryn; Acevedo-Arozena, Abraham; de Silva, Rohan; Greensmith, Linda; Holton, Janice; Hanna, Michael G.; Fisher, Elizabeth M.C.; Fratta, Pietro

2014-01-01

29

Mitochondrial Dysfunction and Decrease in Body Weight of a Transgenic Knock-in Mouse Model for TDP-43.  

Science.gov (United States)

The majority of amyotrophic lateral sclerosis (ALS) cases as well as many patients suffering from frontotemporal lobar dementia (FTLD) with ubiquitinated inclusion bodies show TDP-43 pathology, the protein encoded by the TAR DNA-binding protein (Tardbp) gene. We used recombinase-mediated cassette exchange to introduce an ALS patient cDNA into the mouse Tdp-43 locus. Expression levels of human A315T TDP-43 protein were 300% elevated in heterozygotes, whereas the endogenous mouse Tdp-43 was decreased to 20% of wild type levels as a result of disturbed feedback regulation. Heterozygous TDP-43(A315TKi) mutants lost 10% of their body weight and developed insoluble TDP-43 protein starting as early as 3 months after birth, a pathology that was exacerbated with age. We analyzed the splicing patterns of known Tdp-43 target genes as well as genome-wide gene expression levels in different tissues that indicated mitochondrial dysfunction. In heterozygous mutant animals, we observed a relative decrease in expression of Parkin (Park2) and the fatty acid transporter CD36 along with an increase in fatty acids, HDL cholesterol, and glucose in the blood. As seen in transmission electron microscopy, neuronal cells in motor cortices of TDP-43(A315TKi) animals had abnormal neuronal mitochondrial cristae formation. Motor neurons were reduced to 90%, but only slight motoric impairment was detected. The observed phenotype was interpreted as a predisease model, which might be valuable for the identification of further environmental or genetic triggers of neurodegeneration. PMID:24515116

Stribl, Carola; Samara, Aladin; Trümbach, Dietrich; Peis, Regina; Neumann, Manuela; Fuchs, Helmut; Gailus-Durner, Valerie; Hrab? de Angelis, Martin; Rathkolb, Birgit; Wolf, Eckhard; Beckers, Johannes; Horsch, Marion; Neff, Frauke; Kremmer, Elisabeth; Koob, Sebastian; Reichert, Andreas S; Hans, Wolfgang; Rozman, Jan; Klingenspor, Martin; Aichler, Michaela; Walch, Axel Karl; Becker, Lore; Klopstock, Thomas; Glasl, Lisa; Hölter, Sabine M; Wurst, Wolfgang; Floss, Thomas

2014-04-11

30

Identification of Genetic Modifiers of TDP-43 Neurotoxicity in Drosophila  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Cytosolic aggregation of the nuclear RNA-binding protein TDP-43 is a histopathologic signature of degenerating neurons in amyotrophic lateral sclerosis (ALS), and mutations in the TARDBP gene encoding TDP-43 cause dominantly inherited forms of this condition. To understand the relationship between TDP-43 misregulation and neurotoxicity, we and others have used Drosophila as a model system, in which overexpression of either wild-type TDP-43 or its ALS-associated mutants in neurons is sufficien...

Zhan, Lihong; Hanson, Keith A.; Kim, Sang Hwa; Tare, Apeksha; Tibbetts, Randal S.

2013-01-01

31

Inhibition of TDP-43 Aggregation by Nucleic Acid Binding  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The aggregation of TAR DNA-binding protein (TDP-43) has been shown as a hallmark of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) since 2006. While evidence has suggested that mutation or truncation in TDP-43 influences its aggregation process, nevertheless, the correlation between the TDP-43 aggregation propensity and its binding substrates has not been fully established in TDP-43 proteinopathy. To address this question, we have established a platform based...

Huang, Yi-chen; Lin, Ku-feng; He, Ruei-yu; Tu, Pang-hsien; Koubek, Jiri; Hsu, Yin-chih; Huang, Joseph Jen-tse

2013-01-01

32

TDP-43 Aggregation In Neurodegeneration: Are Stress Granules The Key?  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The RNA-binding protein TDP-43 is strongly linked to neurodegeneration. Not only are mutations in the gene encoding TDP-43 associated with ALS and FTLD, but this protein is also a major constituent of pathological intracellular inclusions in these diseases. Recent studies have significantly expanded our understanding of TDP-43 physiology. TDP-43 is now known to play important roles in neuronal RNA metabolism. It binds to and regulates the splicing and stability of numerous RNAs encoding prote...

Dewey, Colleen M.; Cenik, Basar; Sephton, Chantelle F.; Johnson, Brett A.; Herz, Joachim; Yu, Gang

2012-01-01

33

A90V TDP-43 Variant Results in the Aberrant Localization of TDP-43 In Vitro  

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TAR DNA-binding protein-43 (TDP-43) is a highly conserved, ubiquitously expressed nuclear protein that was recently identified as the disease protein in frontotemporal lobar degeneration with ubiquitin positive inclusions (FTLD-U) and amyotrophic lateral sclerosis (ALS). Pathogenic TDP-43 gene (TARDBP) mutations have been identified in familial ALS kindreds, and here we report a TARDBP variant (A90V) in a FTLD/ALS patient with a family history of dementia. Significantly, A90V is located betwe...

Winton, Matthew J.; Deerlin, Vivianna M.; Kwong, Linda K.; Yuan, Wuxing; Wood, Elisabeth Mccarty; Yu, Chang-en; Schellenberg, Gerard D.; Rademakers, Rosa; Caselli, Richard; Karydas, Anna; Trojanowski, John Q.; Miller, Bruce L.; Lee, Virginia M. -y

2008-01-01

34

Curcumin abolishes mutant TDP-43 induced excitability in a motoneuron-like cellular model of ALS.  

Science.gov (United States)

Mutation of TAR DNA-binding protein-43 (TDP-43) is detected in familial and sporadic amyotrophic lateral sclerosis (FALS and SALS). TDP-43-positive cytoplasmic inclusions are present in both neuron and glia of ALS, although not in mutant Cu/Zn-superoxide dismutase (mSOD1)-related or RNA binding protein Fused in sarcoma (FUS)-related ALS. Previous studies have established that cortical hyper-excitability is common to both FALS and SALS patients. Much of our current understanding of neuron excitability has come from studying the subtype of mSOD1-related ALS. Thus, we evaluated the excitable capability through analyzing properties of action potentials (APs) and voltage-gated sodium (Nav) channels on the cellular model, motoneuron-like cell lines that were steadily transfected with mutant Q331K and wild-type TDP-43. We found that wild-type TDP-43 increased firing frequency of APs, but the presence of mutant Q331K TDP-43 enhanced firing frequency and decreased the threshold of APs to a higher level. Further, we observed that mutant Q331K and wild-type TDP-43 induced more rapid speed of recovery from fast and slow inactivation of Nav channels and resulted in a reduction of voltage dependency of slow inactivation. These results provide evidence for high excitability that resulted from the presence of mutant and wild-type TDP-43, and more toxicity of mutant TDP-43 than wild-type. Other studies suggest that Nav channel activity can be increased directly by different oxidative species and, we have shown previously that oxidative stress and mitochondrial dysfunction occurs simultaneously in the cellular model of mutant TDP-43 and can be ameliorated by dimethoxy curcumin (DMC), a safe and effective antioxidant. In the present study we found that the abnormities of APs and Nav channels were significantly ameliorated when treated with DMC (15?M) for 24h, suggesting a dropping-excitability state. Taken together, mutant Q331K TDP-43 induces high excitability in a motoneuron-like cellular model, and this abnormal state is rescued by DMC which may act through alleviation of oxidative stress and mitochondrial dysfunction. PMID:24785678

Dong, H; Xu, L; Wu, L; Wang, X; Duan, W; Li, H; Li, C

2014-07-11

35

Drosophila TDP-43 dysfunction in glia and muscle cells cause cytological and behavioural phenotypes that characterize ALS and FTLD.  

Science.gov (United States)

Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are neurodegenerative disorders that are characterized by cytoplasmic aggregates and nuclear clearance of TAR DNA-binding protein 43 (TDP-43). Studies in Drosophila, zebrafish and mouse demonstrate that the neuronal dysfunction of TDP-43 is causally related to disease formation. However, TDP-43 aggregates are also observed in glia and muscle cells, which are equally affected in ALS and FTLD; yet, it is unclear whether glia- or muscle-specific dysfunction of TDP-43 contributes to pathogenesis. Here, we show that similar to its human homologue, Drosophila TDP-43, Tar DNA-binding protein homologue (TBPH), is expressed in glia and muscle cells. Muscle-specific knockdown of TBPH causes age-related motor abnormalities, whereas muscle-specific gain of function leads to sarcoplasmic aggregates and nuclear TBPH depletion, which is accompanied by behavioural deficits and premature lethality. TBPH dysfunction in glia cells causes age-related motor deficits and premature lethality. In addition, both loss and gain of Drosophila TDP-43 alter mRNA expression levels of the glutamate transporters Excitatory amino acid transporter 1 (EAAT1) and EAAT2. Taken together, our results demonstrate that both loss and gain of TDP-43 function in muscle and glial cells can lead to cytological and behavioural phenotypes in Drosophila that also characterize ALS and FTLD and identify the glutamate transporters EAAT1/2 as potential direct targets of TDP-43 function. These findings suggest that together with neuronal pathology, glial- and muscle-specific TDP-43 dysfunction may directly contribute to the aetiology and progression of TDP-43-related ALS and FTLD. PMID:23727833

Diaper, Danielle C; Adachi, Yoshitsugu; Lazarou, Luke; Greenstein, Max; Simoes, Fabio A; Di Domenico, Angelique; Solomon, Daniel A; Lowe, Simon; Alsubaie, Rawan; Cheng, Daryl; Buckley, Stephen; Humphrey, Dickon M; Shaw, Christopher E; Hirth, Frank

2013-10-01

36

ALS-Associated TDP-43 Induces Endoplasmic Reticulum Stress, Which Drives Cytoplasmic TDP-43 Accumulation and Stress Granule Formation  

Science.gov (United States)

In amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration, TAR DNA binding protein 43 (TDP-43) accumulates in the cytoplasm of affected neurons and glia, where it associates with stress granules (SGs) and forms large inclusions. SGs form in response to cellular stress, including endoplasmic reticulum (ER) stress, which is induced in both familial and sporadic forms of ALS. Here we demonstrate that pharmacological induction of ER stress causes TDP-43 to accumulate in the cytoplasm, where TDP-43 also associates with SGs. Furthermore, treatment with salubrinal, an inhibitor of dephosphorylation of eukaryotic initiation factor 2-?, a key modulator of ER stress, potentiates ER stress-mediated SG formation. Inclusions of C-terminal fragment TDP-43, reminiscent of disease-pathology, form in close association with ER and Golgi compartments, further indicating the involvement of ER dysfunction in TDP-43-associated disease. Consistent with this notion, over-expression of ALS-linked mutant TDP-43, and to a lesser extent wildtype TDP-43, triggers several ER stress pathways in neuroblastoma cells. Similarly, we found an interaction between the ER chaperone protein disulphide isomerase and TDP-43 in transfected cell lysates and in the spinal cords of mutant A315T TDP-43 transgenic mice. This study provides evidence for ER stress as a pathogenic pathway in TDP-43-mediated disease.

Walker, Adam K.; Soo, Kai Y.; Sundaramoorthy, Vinod; Parakh, Sonam; Ma, Yi; Farg, Manal A.; Wallace, Robyn H.; Crouch, Peter J.; Turner, Bradley J.; Horne, Malcolm K.; Atkin, Julie D.

2013-01-01

37

Aberrant cleavage of TDP-43 enhances aggregation and cellular toxicity  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Inclusions of TAR DNA-binding protein-43 (TDP-43), a nuclear protein that regulates transcription and RNA splicing, are the defining histopathological feature of frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-Us) and sporadic and familial forms of amyotrophic lateral sclerosis (ALS). In ALS and FTLD-U, aggregated, ubiquitinated, and N-terminally truncated TDP-43 can be isolated from brain tissue rich in neuronal and glial cytoplasmic inclusions. The loss of TDP-43 ...

Zhang, Yong-jie; Xu, Ya-fei; Cook, Casey; Gendron, Tania F.; Roettges, Paul; Link, Christopher D.; Lin, Wen-lang; Tong, Jimei; Castanedes-casey, Monica; Ash, Peter; Gass, Jennifer; Rangachari, Vijayaraghavan; Buratti, Emanuele; Baralle, Francisco; Golde, Todd E.

2009-01-01

38

Regulation of Autophagy by Neuropathological Protein TDP-43*  

Digital Repository Infrastructure Vision for European Research (DRIVER)

TDP-43 is a DNA/RNA-binding protein with multicellular functions. As a pathosignature protein of a range of neurodegenerative diseases, TDP-43 is also the major component of the polyubiquitinated inclusions in the pathological cellular samples of these diseases. In normal cells, TDP-43 is processed and degraded by both autophagy and the ubiquitin-proteasome systems. We have found, by microarray hybridization and RT-PCR analyses, that the level of the mRNA encoding the major autophagy componen...

Bose, Jayarama Krishnan; Huang, Chi-chen; Shen, C. -k James

2011-01-01

39

Progranulin and TDP-43: Mechanistic Links and Future Directions  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Loss-of-function mutations in the multifunctional growth factor progranulin (GRN) cause frontotemporal lobar degeneration (FTLD) with TDP-43 protein accumulation. Nuclear TDP-43 protein with key roles in RNA metabolism is also aggregated in amyotrophic lateral sclerosis (ALS), suggesting that ALS and FTLD constitute a broad disease continuum. However, the fact that mutations in GRN are associated with FTLD, while mutations in TDP-43 cause a preferential loss of motor neurons resulting in ALS-...

Kumar-singh, Samir

2011-01-01

40

Does a loss of TDP-43 function cause neurodegeneration?  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract In 2006, TAR-DNA binding protein 43 kDa (TDP-43 was discovered to be in the intracellular aggregates in the degenerating cells in amyotrophic lateral sclerosis (ALS and frontotemporal lobar degeneration (FTLD, two fatal neurodegenerative diseases [1,2]. ALS causes motor neuron degeneration leading to paralysis [3,4]. FTLD causes neuronal degeneration in the frontal and temporal cortices leading to personality changes and a loss of executive function [5]. The discovery triggered a flurry of research activity that led to the discovery of TDP-43 mutations in ALS patients and the widespread presence of TDP-43 aggregates in numerous neurodegenerative diseases. A key question regarding the role of TDP-43 is whether it causes neurotoxicity by a gain of function or a loss of function. The gain-of-function hypothesis has received much attention primarily based on the striking neurodegenerative phenotypes in numerous TDP-43-overexpression models. In this review, I will draw attention to the loss-of-function hypothesis, which postulates that mutant TDP-43 causes neurodegeneration by a loss of function, and in addition, by exerting a dominant-negative effect on the wild-type TDP-43 allele. Furthermore, I will discuss how a loss of function can cause neurodegeneration in patients where TDP-43 is not mutated, review the literature in model systems to discuss how the current data support the loss-of-function mechanism and highlight some key questions for testing this hypothesis in the future.

Xu Zuo-Shang

2012-06-01

 
 
 
 
41

TDP-43 in ALS: stay on target…almost there.  

Science.gov (United States)

ALS is associated with RNA processing impairments involving the RNA-binding protein TDP-43. Pioneering a novel RNA beacon to illuminate RNA trafficking in neurons, Alami et al. (2014) discover a cytoplasmic function for TDP-43, suggesting a new disease mechanism. PMID:24507183

Jovi?i?, Ana; Gitler, Aaron D

2014-02-01

42

TDP-43 is a component of ubiquitin-positive tau-negative inclusions in frontotemporal lobar degeneration and amyotrophic lateral sclerosis  

International Nuclear Information System (INIS)

Ubiquitin-positive tau-negative neuronal cytoplasmic inclusions and dystrophic neurites are common pathological features in frontotemporal lobar degeneration (FTLD) with or without symptoms of motor neuron disease and in amyotrophic lateral sclerosis (ALS). Using biochemical and immunohistochemical analyses, we have identified a TAR DNA-binding protein of 43 kDa (TDP-43), a nuclear factor that functions in regulating transcription and alternative splicing, as a component of these structures in FTLD. Furthermore, skein-like inclusions, neuronal intranuclear inclusions, and glial inclusions in the spinal cord of ALS patients are also positive for TDP-43. Dephosphorylation treatment of the sarkosyl insoluble fraction has shown that abnormal phosphorylation takes place in accumulated TDP-43. The common occurrence of intracellular accumulations of TDP-43 supports the hypothesis that these disorders represent a clinicopathological entity of a single disease, and suggests that they can be newly classified as a proteinopathy of TDP-43

2006-12-22

43

Transposable elements in TDP-43-mediated neurodegenerative disorders.  

Science.gov (United States)

Elevated expression of specific transposable elements (TEs) has been observed in several neurodegenerative disorders. TEs also can be active during normal neurogenesis. By mining a series of deep sequencing datasets of protein-RNA interactions and of gene expression profiles, we uncovered extensive binding of TE transcripts to TDP-43, an RNA-binding protein central to amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Second, we find that association between TDP-43 and many of its TE targets is reduced in FTLD patients. Third, we discovered that a large fraction of the TEs to which TDP-43 binds become de-repressed in mouse TDP-43 disease models. We propose the hypothesis that TE mis-regulation contributes to TDP-43 related neurodegenerative diseases. PMID:22957047

Li, Wanhe; Jin, Ying; Prazak, Lisa; Hammell, Molly; Dubnau, Josh

2012-01-01

44

Multiplex SILAC analysis of a cellular TDP-43 proteinopathy model reveals protein inclusions associated with SUMOylation and diverse polyubiquitin chains.  

Science.gov (United States)

Transactive response (TAR) DNA-binding protein 43 (TDP-43) is a major protein component within ubiquitin-positive inclusions of frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Although TDP-43 is a nuclear DNA/RNA-binding protein, in pathological conditions, TDP-43 has been reported to redistribute to the cytoplasm where it is cleaved and forms insoluble, ubiquitinated, and phosphorylated inclusions. Here we present a cellular model in which full-length human TDP-43 or a splicing isoform (TDP-S6) that lacks the C terminus is overexpressed in a human cell line and mouse primary neurons. Whereas recombinant and endogenous TDP-43 was primarily localized in the nucleus, the shorter TDP-S6 formed highly insoluble cytoplasmic and nuclear inclusions reminiscent of disease-specific pathology. Western blot analysis of detergent-insoluble extracts showed an increase in high molecular weight immunoreactive species for TDP-S6 compared with TDP-43, consistent with ubiquitination or ubiquitin-like modifications. We used a multiplex stable isotope labeling with amino acids in cell culture approach to compare the detergent-insoluble proteome from mock-, TDP-43-, and TDP-S6-transfected cells. TDP-S6 overexpression caused a concomitant increase in both ubiquitin (Ub) and the small Ub-like modifier-2/3 (SUMO-2/3) within the insoluble proteome. Similarly, full-length TDP-43 overexpression also resulted in the elevation of SUMO-2/3. Immunofluorescence showed strong co-localization of endogenous Ub with both cytoplasmic and nuclear TDP-S6 inclusions, whereas SUMO-2/3 was co-localized mainly with the nuclear inclusions. Quantitative mass spectrometry further revealed that mixed Lys-48 and Lys-63 polyUb linkages were associated with the TDP insoluble fractions. Together our data indicate that expression of a TDP-43 splice variant lacking a C terminus recapitulates many of the cellular and biochemical features associated with disease pathology and that the interplay of ubiquitination and SUMOylation may have an important role in TDP-43 regulation. PMID:20047951

Seyfried, Nicholas T; Gozal, Yair M; Dammer, Eric B; Xia, Qiangwei; Duong, Duc M; Cheng, Dongmei; Lah, James J; Levey, Allan I; Peng, Junmin

2010-04-01

45

An ALS-associated mutation affecting TDP-43 enhances protein aggregation, fibril formation and neurotoxicity  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Mutations in TARDBP, encoding TAR DNA-binding protein-43 (TDP-43), are associated with TDP-43 proteinopathies, including amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). We compared wild-type TDP-43 and an ALS-associated mutant TDP-43 in vitro and in vivo. The A315T mutant enhances neurotoxicity and the formation of aberrant TDP-43 species, including protease-resistant fragments. The C terminus of TDP-43 shows sequence similarity to prion proteins. Synthetic p...

Guo, Weirui; Chen, Yanbo; Zhou, Xiaohong; Kar, Amar; Ray, Payal; Chen, Xiaoping; Rao, Elizabeth J.; Yang, Mengxue; Ye, Haihong; Zhu, Li; Liu, Jianghong; Xu, Meng; Yang, Yanlian; Wang, Chen; Zhang, David

2011-01-01

46

ALS-associated mutations in TDP-43 increase its stability and promote TDP-43 complexes with FUS/TLS  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Dominant mutations in two functionally related DNA/RNA-binding proteins, trans-activating response region (TAR) DNA-binding protein with a molecular mass of 43 KDa (TDP-43) and fused in sarcoma/translocation in liposarcoma (FUS/TLS), cause an inherited form of ALS that is accompanied by nuclear and cytoplasmic aggregates containing TDP-43 or FUS/TLS. Using isogenic cell lines expressing wild-type or ALS-linked TDP-43 mutants and fibroblasts from a human patient, pulse-chase radiolabeling of n...

Ling, Shuo-chien; Albuquerque, Claudio P.; Han, Joo Seok; Lagier-tourenne, Clotilde; Tokunaga, Seiya; Zhou, Huilin; Cleveland, Don W.

2010-01-01

47

TDP-43 promotes microRNA biogenesis as a component of the Drosha and Dicer complexes  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Although aberrant microRNA (miRNA) expression is linked to human diseases including cancer, the mechanisms that regulate the expression of each individual miRNA remain largely unknown. TAR DNA-binding protein-43 (TDP-43) is homologous to the heterogeneous nuclear ribonucleoproteins (hnRNPs), which are involved in RNA processing, and its abnormal cellular distribution is a key feature of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD), two neurodegenerative dis...

Kawahara, Yukio; Mieda-sato, Ai

2012-01-01

48

TDP-1/TDP-43 Regulates Stress Signaling and Age-Dependent Proteotoxicity in Caenorhabditis elegans  

Digital Repository Infrastructure Vision for European Research (DRIVER)

TDP-43 is a multifunctional nucleic acid binding protein linked to several neurodegenerative diseases including Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Dementia. To learn more about the normal biological and abnormal pathological role of this protein, we turned to Caenorhabditis elegans and its orthologue TDP-1. We report that TDP-1 functions in the Insulin/IGF pathway to regulate longevity and the oxidative stress response downstream from the forkhead transcription factor DAF-...

Vaccaro, Alexandra; Tauffenberger, Arnaud; Ash, Peter E. A.; Carlomagno, Yari; Petrucelli, Leonard; Parker, J. Alex

2012-01-01

49

Therapeutic modulation of eIF2? phosphorylation rescues TDP-43 toxicity in amyotrophic lateral sclerosis disease models.  

Science.gov (United States)

Amyotrophic lateral sclerosis (ALS) is a fatal, late-onset neurodegenerative disease primarily affecting motor neurons. A unifying feature of many proteins associated with ALS, including TDP-43 and ataxin-2, is that they localize to stress granules. Unexpectedly, we found that genes that modulate stress granules are strong modifiers of TDP-43 toxicity in Saccharomyces cerevisiae and Drosophila melanogaster. eIF2? phosphorylation is upregulated by TDP-43 toxicity in flies, and TDP-43 interacts with a central stress granule component, polyA-binding protein (PABP). In human ALS spinal cord neurons, PABP accumulates abnormally, suggesting that prolonged stress granule dysfunction may contribute to pathogenesis. We investigated the efficacy of a small molecule inhibitor of eIF2? phosphorylation in ALS models. Treatment with this inhibitor mitigated TDP-43 toxicity in flies and mammalian neurons. These findings indicate that the dysfunction induced by prolonged stress granule formation might contribute directly to ALS and that compounds that mitigate this process may represent a novel therapeutic approach. PMID:24336168

Kim, Hyung-Jun; Raphael, Alya R; LaDow, Eva S; McGurk, Leeanne; Weber, Ross A; Trojanowski, John Q; Lee, Virginia M-Y; Finkbeiner, Steven; Gitler, Aaron D; Bonini, Nancy M

2014-02-01

50

Phosphorylation promotes neurotoxicity in a C. elegans model of TDP-43 proteinopathy  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Neurodegenerative disorders characterized by neuronal and glial lesions containing aggregated pathological TDP-43 protein in the cytoplasm, nucleus, or neurites are collectively referred to as TDP-43 proteinopathies. Lesions containing aggregated TDP-43 protein are a hallmark of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with ubiquitinated inclusions (FTLD-U). In addition, mutations in human TDP-43 cause ALS. We have developed a C. elegans model of TDP-43 protei...

Liachko, Nicole F.; Guthrie, Chris R.; Kraemer, Brian C.

2010-01-01

51

Interaction with Polyglutamine Aggregates Reveals a Q/N-rich Domain in TDP-43*  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The identification of pathologic TDP-43 aggregates in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration, followed by the discovery of dominantly inherited point mutations in TDP-43 in familial ALS, have been critical insights into the mechanism of these untreatable neurodegenerative diseases. However, the biochemical basis of TDP-43 aggregation and the mechanism of how mutations in TDP-43 lead to disease remain enigmatic. In efforts to understand how TDP-43 alters its ...

Fuentealba, Rodrigo A.; Udan, Maria; Bell, Shaughn; Wegorzewska, Iga; Shao, Jieya; Diamond, Marc I.; Weihl, Conrad C.; Baloh, Robert H.

2010-01-01

52

Inhibition of RNA lariat debranching enzyme suppresses TDP-43 toxicity in ALS disease models  

Digital Repository Infrastructure Vision for European Research (DRIVER)

ALS is a devastating neurodegenerative disease primarily affecting motor neurons. Mutations in TDP-43 cause some forms of the disease, and cytoplasmic TDP-43 aggregates accumulate in degenerating neurons of most ALS patients. Thus, strategies aimed at targeting the toxicity of cytoplasmic TDP-43 aggregates may be effective. Here we report results from two genome-wide loss-of-function TDP-43 toxicity suppressor screens in yeast. The strongest suppressor of TDP-43 toxicity was deletion of Dbr1,...

Armakola, Maria; Higgins, Matthew J.; Figley, Matthew D.; Barmada, Sami J.; Scarborough, Emily A.; Diaz, Zamia; Fang, Xiaodong; Shorter, James; Krogan, Nevan J.; Finkbeiner, Steven; Farese, Robert V.; Gitler, Aaron D.

2012-01-01

53

Wild-type human TDP-43 expression causes TDP-43 phosphorylation, mitochondrial aggregation, motor deficits and early mortality in transgenic mice  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Transactivation response DNA-binding protein 43 (TDP-43) is a principal component of ubiquitinated inclusions in frontotemporal lobar degeneration with ubiquitin-positive inclusions and in amyotrophic lateral sclerosis (ALS). Mutations in TARDBP, the gene encoding TDP-43, are associated with sporadic and familial ALS, yet multiple neurodegenerative diseases exhibit TDP-43 pathology without known TARDBP mutations. While TDP-43 has been ascribed a number of roles in normal biology, including mR...

Xu, Ya-fei; Gendron, Tania F.; Zhang, Yong-jie; Lin, Wen-lang; D’alton, Simon; Sheng, Hong; Casey, Monica Castanedes; Tong, Jimei; Knight, Joshua; Yu, Xin; Rademakers, Rosa; Boylan, Kevin; Hutton, Mike; Mcgowan, Eileen; Dickson, Dennis W.

2010-01-01

54

TDP-43 Is a Transcriptional Repressor: THE TESTIS-SPECIFIC MOUSE acrv1 GENE IS A TDP-43 TARGET IN VIVO*  

Digital Repository Infrastructure Vision for European Research (DRIVER)

TDP-43 is an evolutionarily conserved ubiquitously expressed DNA/RNA-binding protein. Although recent studies have shown its association with a variety of neurodegenerative disorders, the function of TDP-43 remains poorly understood. Here we address TDP-43 function using spermatogenesis as a model system. We previously showed that TDP-43 binds to the testis-specific mouse acrv1 gene promoter in vitro via two GTGTGT-motifs and that mutation of these motifs led to premature transcription in spe...

Lalmansingh, Avin S.; Urekar, Craig J.; Reddi, Prabhakara P.

2011-01-01

55

"Structural characterization of the minimal segment of TDP-43 competent for aggregation".  

Science.gov (United States)

TDP-43 is a nuclear protein whose abnormal aggregates are implicated in ALS and FTLD. Recently, an Asn/Gln rich C-terminal segment of TDP-43 has been shown to produce aggregation in vitro and reproduce most of the protein's pathological hallmarks in cells, but little is known about this segment's structure. Here, CD and 2D heteronuclear NMR spectroscopies provide evidence that peptides corresponding to the wild type and mutated sequences of this segment adopt chiefly disordered conformations that, in the case of the wild type sequence, spontaneously forms a ?-sheet rich oligomer. Moreover, MD simulation provides evidence for a structure consisting of two ?-strands and a well-defined, yet non-canonical structural element. Furthermore, MD simulations of four pathological mutations (Q343R, N345K, G348V and N352S) occurring in this segment predict that all of them could affect this region's structure. In particular, the Q343R variant tends to stabilize disordered conformers, N345K permits the formation of longer, more stable ?-strands, and G348V tends to shorten and destabilize them. Finally, N352S acts to alter the ?-stand register and when S352 is phosphorylated, it induces partial unfolding. Our results provide a better understanding of TDP-43 aggregation process and will be useful to design effectors capable to modulate its progression. PMID:24440310

Mompeán, Miguel; Buratti, Emanuele; Guarnaccia, Corrado; Brito, Rui M M; Chakrabartty, Avijit; Baralle, Francisco E; Laurents, Douglas V

2014-03-01

56

TDP-43 accumulation in IBM muscle suggests a common pathogenic mechanism with Frontotemporal dementia  

Digital Repository Infrastructure Vision for European Research (DRIVER)

TDP-43 is found in ubiquitinated inclusions (UBIs) in some frontotemporal dementias (FTD-U). One form of FTD-U, due to mutations in VCP, occurs with an inclusion body myopathy (IBMPFD). Since IBMPFD brain has TDP-43 in UBIs, we looked for TDP-43 inclusions in IBMPFD muscle. In normal muscle TDP-43 is present in nuclei. In IBMPFD muscle TDP-43 is additionally present as large inclusions within UBIs in muscle cytoplasm. TDP-43 inclusions were also found in 78% of sIBM muscles. In IBMPFD and sIB...

Weihl, Conrad C.; Temiz, Peyker; Miller, Sara E.; Watts, Giles; Smith, Charles; Forman, Mark; Hanson, Phyllis I.; Kimonis, Virginia; Pestronk, Alan

2008-01-01

57

Partial loss of TDP-43 function causes phenotypes of amyotrophic lateral sclerosis.  

Science.gov (United States)

Amyotrophic lateral sclerosis (ALS) is a fatal neurological disease that causes motor neuron degeneration, progressive motor dysfunction, paralysis, and death. Although multiple causes have been identified for this disease, >95% of ALS cases show aggregation of transactive response DNA binding protein (TDP-43) accompanied by its nuclear depletion. Therefore, the TDP-43 pathology may be a converging point in the pathogenesis that originates from various initial triggers. The aggregation is thought to result from TDP-43 misfolding, which could generate cellular toxicity. However, the aggregation as well as the nuclear depletion could also lead to a partial loss of TDP-43 function or TDP-43 dysfunction. To investigate the impact of TDP-43 dysfunction, we generated a transgenic mouse model for a partial loss of TDP-43 function using transgenic RNAi. These mice show ubiquitous transgene expression and TDP-43 knockdown in both the periphery and the central nervous system (CNS). Strikingly, these mice develop progressive neurodegeneration prominently in cortical layer V and spinal ventral horn, motor dysfunction, paralysis, and death. Furthermore, examination of splicing patterns of TDP-43 target genes in human ALS revealed changes consistent with TDP-43 dysfunction. These results suggest that the CNS, particularly motor neurons, possess a heightened vulnerability to TDP-43 dysfunction. Additionally, because TDP-43 knockdown predominantly occur in astrocytes in the spinal cord of these mice, our results suggest that TDP-43 dysfunction in astrocytes is an important driver for motor neuron degeneration and clinical phenotypes of ALS. PMID:24616503

Yang, Chunxing; Wang, Hongyan; Qiao, Tao; Yang, Bin; Aliaga, Leonardo; Qiu, Linghua; Tan, Weijia; Salameh, Johnny; McKenna-Yasek, Diane M; Smith, Thomas; Peng, Lingtao; Moore, Melissa J; Brown, Robert H; Cai, Huaibin; Xu, Zuoshang

2014-03-25

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A Nonsense Mutation in Mouse Tardbp Affects TDP43 Alternative Splicing Activity and Causes Limb-Clasping and Body Tone Defects  

Science.gov (United States)

Mutations in TARDBP, encoding Tar DNA binding protein-43 (TDP43), cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Attempts to model TDP43 dysfunction in mice have used knockouts or transgenic overexpressors, which have revealed the difficulties of manipulating TDP43, whose level is tightly controlled by auto-regulation. In a complementary approach, to create useful mouse models for the dissection of TDP43 function and pathology, we have identified a nonsense mutation in the endogenous mouse Tardbp gene through screening an N-ethyl-N-nitrosourea (ENU) mutant mouse archive. The mutation is predicted to cause a Q101X truncation in TDP43. We have characterised TardbpQ101X mice to investigate this mutation in perturbing TDP43 biology at endogenous expression levels. We found the TardbpQ101X mutation is homozygous embryonic lethal, highlighting the importance of TDP43 in early development. Heterozygotes (Tardbp+/Q101X) have abnormal levels of mutant transcript, but we find no evidence of the truncated protein and mice have similar full-length TDP43 protein levels as wildtype littermates. Nevertheless, Tardbp+/Q101X mice have abnormal alternative splicing of downstream gene targets, and limb-clasp and body tone phenotypes. Thus the nonsense mutation in Tardbp causes a mild loss-of-function phenotype and behavioural assessment suggests underlying neurological abnormalities. Due to the role of TDP43 in ALS, we investigated potential interactions with another known causative gene, mutant superoxide dismutase 1 (SOD1). Tardbp+/Q101X mice were crossed with the SOD1G93Adl transgenic mouse model of ALS. Behavioural and physiological assessment did not reveal modifying effects on the progression of ALS-like symptoms in the double mutant progeny from this cross. In summary, the TardbpQ101X mutant mice are a useful tool for the dissection of TDP43 protein regulation, effects on splicing, embryonic development and neuromuscular phenotypes. These mice are freely available to the community.

Fratta, Pietro; de Oliveira, Hugo M.; Kent, Rosie; Phatak, Vinaya; Brandner, Sebastian; Blanco, Gonzalo; Greensmith, Linda; Acevedo-Arozena, Abraham; Fisher, Elizabeth M. C.

2014-01-01

59

Heat-shock protein dysregulation is associated with functional and pathological TDP-43 aggregation  

Science.gov (United States)

Conformational disorders are involved in various neurodegenerative diseases. Reactive oxygen species (ROS) are the major contributors to neurodegenerative disease; however, ROS that affect the structural changes in misfolded disease proteins have yet to be well characterized. Here we demonstrate that the intrinsic propensity of TDP-43 to aggregate drives the assembly of TDP-43-positive stress granules and soluble toxic TDP-43 oligomers in response to a ROS insult via a disulfide crosslinking-independent mechanism. Notably, ROS-induced TDP-43 protein assembly correlates with the dynamics of certain TDP-43-associated chaperones. The heat-shock protein (HSP)-90 inhibitor 17-AAG prevents ROS-induced TDP-43 aggregation, alters the type of TDP-43 multimers and reduces the severity of pathological TDP-43 inclusions. In summary, our study suggests that a common mechanism could be involved in the pathogenesis of conformational diseases that result from HSP dysregulation.

Chang, Hsiang-Yu; Hou, Shin-Chen; Way, Tzong-Der; Wong, Chi-Huey; Wang, I.-Fan

2013-11-01

60

TDP-43 accumulation in IBM muscle suggests a common pathogenic mechanism with Frontotemporal dementia  

Science.gov (United States)

TDP-43 is found in ubiquitinated inclusions (UBIs) in some frontotemporal dementias (FTD-U). One form of FTD-U, due to mutations in VCP, occurs with an inclusion body myopathy (IBMPFD). Since IBMPFD brain has TDP-43 in UBIs, we looked for TDP-43 inclusions in IBMPFD muscle. In normal muscle TDP-43 is present in nuclei. In IBMPFD muscle TDP-43 is additionally present as large inclusions within UBIs in muscle cytoplasm. TDP-43 inclusions were also found in 78% of sIBM muscles. In IBMPFD and sIBM muscle TDP-43 migrated with an additional band on immunoblot similar to that reported in FTD-U brains. This study adds sIBM and hereditary inclusion body myopathies to the growing list of TDP-43 positive inclusion diseases.

Weihl, Conrad C.; Temiz, Peyker; Miller, Sara E.; Watts, Giles; Smith, Charles; Forman, Mark; Hanson, Phyllis I.; Kimonis, Virginia; Pestronk, Alan

2008-01-01

 
 
 
 
61

Regulation of nuclear TDP-43 by NR2A-containing NMDA receptors and PTEN  

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The dysfunction of TAR DNA-binding protein-43 (TDP-43) is implicated in neurodegenerative diseases. However, the function of TDP-43 is not fully elucidated. Here we show that the protein level of endogenous TDP-43 in the nucleus is increased in mouse cortical neurons in the early stages, but return to basal level in the later stages after glutamate accumulation-induced injury. The elevation of TDP-43 results from a downregulation of phosphatase and tensin homolog (PTEN). We further demonstrat...

Zheng, Mei; Liao, Mingxia; Cui, Tianyuan; Tian, Honglin; Fan, Dong-sheng; Wan, Qi

2012-01-01

62

Rapamycin Rescues TDP-43 Mislocalization and the Associated Low Molecular Mass Neurofilament Instability  

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TDP-43 is a nuclear protein involved in exon skipping and alternative splicing. Recently, TDP-43 has been identified as the pathological signature protein in frontotemporal lobar degeneration with ubiquitin-positive inclusions and in amyotrophic lateral sclerosis. In addition, TDP-43-positive inclusions are present in Parkinson disease, dementia with Lewy bodies, and 30% of Alzheimer disease cases. Pathological TDP-43 is redistributed from the nucleus to the cytoplasm, where it accumulates. A...

Caccamo, Antonella; Majumder, Smita; Deng, Janice J.; Bai, Yidong; Thornton, Fiona B.; Oddo, Salvatore

2009-01-01

63

TDP-43 Phosphorylation by casein kinase I? promotes oligomerization and enhances toxicity in vivo.  

Science.gov (United States)

Dominant mutations in transactive response DNA-binding protein-43 (TDP-43) cause amyotrophic lateral sclerosis. TDP-43 inclusions occur in neurons, glia and muscle in this disease and in sporadic and inherited forms of frontotemporal lobar degeneration. Cytoplasmic localization, cleavage, aggregation and phosphorylation of TDP-43 at the Ser409/410 epitope have been associated with disease pathogenesis. TDP-43 aggregation is not a common feature of mouse models of TDP-43 proteinopathy, and TDP-43 is generally not thought to acquire an amyloid conformation or form fibrils. A number of putative TDP-43 kinases have been identified, but whether any of these functions to regulate TDP-43 phosphorylation or toxicity in vivo is not known. Here, we demonstrate that human TDP-43(Q331K) undergoes cytoplasmic localization and aggregates when misexpressed in Drosophila when compared with wild-type and M337V forms. Coexpression of Q331K with doubletime (DBT), the fly homolog of casein kinase I? (CKI?), enhances toxicity. There is at best modest basal phosphorylation of misexpressed human TDP-43 in Drosophila, but coexpression with DBT increases Ser409/410 phosphorylation of all TDP-43 isoforms tested. Phosphorylation of TDP-43 in the fly is specific for DBT, as it is not observed using the validated tau kinases GSK-3?, PAR-1/MARK2 or CDK5. Coexpression of DBT with TDP-43(Q331K) enhances the formation of high-molecular weight oligomeric species coincident with enhanced toxicity, and treatment of recombinant oligomeric TDP-43 with rat CKI strongly enhances its toxicity in mammalian cell culture. These data identify CKI? as a potent TDP-43 kinase in vivo and implicate oligomeric species as the toxic entities in TDP-43 proteinopathies. PMID:24105464

Choksi, Darshana K; Roy, Bidisha; Chatterjee, Shreyasi; Yusuff, Tanzeen; Bakhoum, Mathieu F; Sengupta, Urmi; Ambegaokar, Suren; Kayed, Rakez; Jackson, George R

2014-02-15

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Ubiquilin Modifies TDP-43 Toxicity in a Drosophila Model of Amyotrophic Lateral Sclerosis (ALS)*  

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TDP-43 (43-kDa TAR DNA-binding protein) is a major constituent of ubiquitin-positive cytosolic aggregates present in neurons of patients with amyotrophic lateral sclerosis (ALS) and ubiquitin-positive fronto-temporal lobar degeneration (FTLD-U). Inherited mutations in TDP-43 have been linked to familial forms of ALS, indicating a key role for TDP-43 in disease pathogenesis. Here, we describe a Drosophila melanogaster model of TDP-43 proteinopathy. Expression of wild-type human TDP-43 protein ...

Hanson, Keith A.; Kim, Sang Hwa; Wassarman, David A.; Tibbetts, Randal S.

2010-01-01

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Sustained Expression of TDP-43 and FUS in Motor Neurons in Rodent's Lifetime  

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Full Text Available TAR DNA-binding protein (TDP-43 and fused in sarcoma (FUS are two highly conserved ribonucleoproteins. Pathogenic mutations of the TDP-43 or the FUS gene are all linked to amyotrophic lateral sclerosis (ALS that is characterized by progressive degeneration of motor neurons. To better understand the correlation of ALS disease genes with the selectivity of chronic motor neuron degeneration, we examined the longitudinal expression of the TDP-43 and the FUS genes in C57BL6 mice and in Sprague-Dawley rats. TDP-43 and FUS were robustly and ubiquitously expressed in the postnatal mice and rats, but were markedly decreased in the adult rodents. In adulthood, TDP-43 and FUS proteins were even undetectable in peripheral organs including skeletal muscles, liver, and kidney, but were constantly expressed at substantial levels in the central nervous system. Motor neurons expressed the TDP-43 and the FUS genes at robust levels throughout rodent's lifetime. Moreover, TDP-43 and FUS were accumulated in the cytoplasm of motor neurons in aged animals. Our findings suggest that TDP-43 and FUS play an important role in development and that constant and robust expression of the genes in motor neurons may render the neurons vulnerable to pathogenic mutation of the TDP-43 or the FUS gene. To faithfully model the pathology of TDP-43- or FUS gene mutations in rodents, we must replicate the expression patterns of the TDP-43 and the FUS gene in animals.

Cao Huang, Pedro Yuxing Xia, Hongxia Zhou

2010-01-01

66

Frontotemporal dementia and amyotrophic lateral sclerosis-associated disease protein TDP-43 promotes dendritic branching  

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Full Text Available Abstract Background TDP-43 is an evolutionarily conserved RNA-binding protein implicated in the pathogenesis of frontotemporal dementia (FTD, sporadic and familial amyotrophic lateral sclerosis (ALS, and possibly other neurodegenerative diseases. In diseased neurons, TDP-43 is depleted in the nucleus, suggesting a loss-of-function pathogenic mechanism. However, the normal function of TDP-43 in postmitotic neurons is largely unknown. Results Here we demonstrate that overexpression of Drosophila TDP-43 (dTDP-43 in vivo significantly increases dendritic branching of sensory neurons in Drosophila larvae. Loss of dTDP-43 function, either in a genetic null mutant or through RNAi knockdown, decreased dendritic branching. Further genetic analysis demonstrated a cell-autonomous role for dTDP-43 in dendrite formation. Moreover, human TDP-43 (hTDP-43 promoted dendritic branching in Drosophila neurons, and this function was attenuated by mutations associated with ALS. Conclusion These findings reveal an essential role for TDP-43 in dendritic structural integrity, supporting the notion that loss of normal TDP-43 function in diseased neurons may compromise neuronal connectivity before neuronal cell loss in FTD and ALS.

Lu Yubing

2009-09-01

67

Does a loss of TDP-43 function cause neurodegeneration?  

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Abstract In 2006, TAR-DNA binding protein 43 kDa (TDP-43) was discovered to be in the intracellular aggregates in the degenerating cells in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD), two fatal neurodegenerative diseases [1,2]. ALS causes motor neuron degeneration leading to paralysis [3,4]. FTLD causes neuronal degeneration in the frontal and temporal cortices leading to personality changes and a loss of executive function [5]. The discovery tr...

Xu Zuo-Shang

2012-01-01

68

Disease causing mutants of TDP-43 nucleic acid binding domains are resistant to aggregation and have increased stability and half-life.  

Science.gov (United States)

Over the last two decades many secrets of the age-related human neural proteinopathies have been revealed. A common feature of these diseases is abnormal, and possibly pathogenic, aggregation of specific proteins in the effected tissue often resulting from inherent or decreased structural stability. An archetype example of this is superoxide dismutase-1, the first genetic factor to be linked with amyotrophic lateral sclerosis (ALS). Mutant or posttranslationally modified TAR DNA binding protein-32 (TDP-43) is also strongly associated with ALS and an increasingly large number of other neurodegenerative diseases, including frontotemporal lobar degeneration (FTLD). Cytoplasmic mislocalization and elevated half-life is a characteristic of mutant TDP-43. Furthermore, patient age at the onset of disease symptoms shows a good inverse correlation with mutant TDP-43 half-life. Here we show that ALS and FTLD-associated TDP-43 mutations in the central nucleic acid binding domains lead to elevated half-life and this is commensurate with increased thermal stability and inhibition of aggregation. It is achieved without impact on secondary, tertiary, or quaternary structure. We propose that tighter structural cohesion contributes to reduced protein turnover, increasingly abnormal proteostasis and, ultimately, faster onset of disease symptoms. These results contrast our perception of neurodegenerative diseases as misfolded proteinopathies and delineate a novel path from the molecular characteristics of mutant TDP-43 to aberrant cellular effects and patient phenotype. PMID:24591609

Austin, James A; Wright, Gareth S A; Watanabe, Seiji; Grossmann, J Günter; Antonyuk, Svetlana V; Yamanaka, Koji; Hasnain, S Samar

2014-03-18

69

Mimicking aspects of frontotemporal lobar degeneration and Lou Gehrig's disease in rats via TDP-43 overexpression.  

Science.gov (United States)

Since the discovery of neuropathological lesions made of TDP-43 and ubiquitin proteins in cases of frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS), there is a burst of effort on finding related familial mutations and developing animal models. We used an adeno-associated virus (AAV) vector for human TDP-43 expression targeted to the substantia nigra (SN) of rats. Though TDP-43 was expressed mainly in neuronal nuclei as expected, it was also expressed in the cytoplasm, and dotted along the plasma membrane of neurons. Cytoplasmic staining was both diffuse and granular, indicative of preinclusion lesions, over 4 weeks. Ubiquitin deposited in the cytoplasm, specifically in the TDP-43 group, and staining for microglia was increased dose-dependently by 1-2 logs in the TDP-43 group, while neurons were selectively obliterated. Neuronal death induced by TDP-43 was pyknotic and apoptotic. TDP-43 gene transfer caused loss of dopaminergic neurons in the SN and their axons in the striatum. Behavioral motor dysfunction resulted after TDP-43 gene transfer that was vector dose-dependent and progressive over time. The cytoplasmic expression, ubiquitination, and neurodegeneration mimicked features of the TDP-43 diseases, and the gliosis, apoptosis, and motor impairment may also be relevant to TDP-43 disease forms involving nigrostriatal degeneration. PMID:19223871

Tatom, Jason B; Wang, David B; Dayton, Robert D; Skalli, Omar; Hutton, Michael L; Dickson, Dennis W; Klein, Ronald L

2009-04-01

70

Tar DNA Binding Protein-43 (TDP-43) Associates with Stress Granules: Analysis of Cultured Cells and Pathological Brain Tissue  

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Tar DNA Binding Protein-43 (TDP-43) is a principle component of inclusions in many cases of frontotemporal lobar degeneration (FTLD-U) and amyotrophic lateral sclerosis (ALS). TDP-43 resides predominantly in the nucleus, but in affected areas of ALS and FTLD-U central nervous system, TDP-43 is aberrantly processed and forms cytoplasmic inclusions. The mechanisms governing TDP-43 inclusion formation are poorly understood. Increasing evidence indicates that TDP-43 regulates mRNA metabolism by i...

Liu-yesucevitz, Liqun; Bilgutay, Aylin; Zhang, Yong-jie; Vanderwyde, Tara; Citro, Allison; Mehta, Tapan; Zaarur, Nava; Mckee, Ann; Bowser, Robert; Sherman, Michael; Petrucelli, Leonard; Wolozin, Benjamin

2010-01-01

71

Mimicking Aspects of Frontotemporal Lobar Degeneration and Lou Gehrig's Disease in Rats via TDP-43 Overexpression  

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Since the discovery of neuropathological lesions made of TDP-43 and ubiquitin proteins in cases of frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS), there is a burst of effort on finding related familial mutations and developing animal models. We used an adeno-associated virus (AAV) vector for human TDP-43 expression targeted to the substantia nigra (SN) of rats. Though TDP-43 was expressed mainly in neuronal nuclei as expected, it was also expressed in the cyt...

Tatom, Jason B.; Wang, David B.; Dayton, Robert D.; Skalli, Omar; Hutton, Michael L.; Dickson, Dennis W.; Klein, Ronald L.

2009-01-01

72

Cytoplasmic Accumulation and Aggregation of TDP-43 upon Proteasome Inhibition in Cultured Neurons  

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Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are characterized by intraneuronal deposition of the nuclear TAR DNA-binding protein 43 (TDP-43) caused by unknown mechanisms. Here, we studied TDP-43 in primary neurons under different stress conditions and found that only proteasome inhibition by MG-132 or lactacystin could induce significant cytoplasmic accumulation of TDP-43, a histopathological hallmark in disease. This cytoplasmic accumulation was accompani...

Eersel, Janet; Ke, Yazi D.; Gladbach, Amadeus; Bi, Mian; Go?tz, Ju?rgen; Kril, Jillian J.; Ittner, Lars M.

2011-01-01

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TDP-43 A315T Mutation in Familial Motor Neuron Disease  

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To identify novel causes of familial neurodegenerative diseases, we extended our previous studies of TAR DNA-binding protein 43 (TDP-43) proteinopathies to investigate TDP-43 as a candidate gene in familial cases of motor neuron disease. Sequencing of the TDP-43 gene led to the identification of a novel missense mutation, Ala-315-Thr, which segregates with all affected members of an autosomal dominant motor neuron disease family. The mutation was not found in 1,505 healthy control subjects. T...

Gitcho, Michael A.; Baloh, Robert H.; Chakraverty, Sumi; Mayo, Kevin; Norton, Joanne B.; Levitch, Denise; Hatanpaa, Kimmo J.; White, Charles L.; Bigio, Eileen H.; Caselli, Richard; Baker, Matt; Al-lozi, Muhammad T.; Morris, John C.; Pestronk, Alan; Rademakers, Rosa

2008-01-01

74

Review: Transactive response DNA-binding protein 43 (TDP-43): mechanisms of neurodegeneration  

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Since the identification of phosphorylated and truncated transactive response DNA-binding protein 43 (TDP-43) as a primary component of ubiquitinated inclusions in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with ubiquitin-positive inclusions, and the discovery that mutations in the TDP-43 gene cause ALS, much effort has been directed towards establishing how TDP-43 contributes to the development of neurodegeneration. Although few in vivo models are presently ava...

Gendron, T. F.; Josephs, K. A.; Petrucelli, L.

2010-01-01

75

TDP-43 regulates retinoblastoma protein phosphorylation through the repression of cyclin-dependent kinase 6 expression  

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TDP-43 (for TAR DNA binding protein) is a highly conserved heterogeneous nuclear ribonucleoprotein (hnRNP) involved in specific pre-mRNA splicing and transcription events. TDP-43 recently has been identified as the main component of cytoplasmic inclusions in frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS), two neurodegenerative disorders. The cellular role of this protein remains to be identified. Here, we show that loss of TDP-43 results in dysmorphic nuclear...

Ayala, Youhna M.; Misteli, Tom; Baralle, Francisco E.

2008-01-01

76

TDP-43 Potentiates Alpha-synuclein Toxicity to Dopaminergic Neurons in Transgenic Mice  

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TDP-43 and ?-synuclein are two disease proteins involved in a wide range of neurodegenerative diseases. While TDP-43 proteinopathy is considered a pathologic hallmark of sporadic amyotrophic lateral sclerosis and frontotemporal lobe degeneration, ?-synuclein is a major component of Lewy body characteristic of Parkinson's disease. Intriguingly, TDP-43 proteinopathy also coexists with Lewy body and with synucleinopathy in certain disease conditions. Here we reported the effects of ...

Tian Tian, Cao Huang

2011-01-01

77

Global analysis of TDP-43 interacting proteins reveals strong association with RNA splicing and translation machinery  

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TDP-43 is a highly conserved and ubiquitously expressed member of the heterogeneous nuclear ribonucleoprotein (hnRNP) family of proteins. Recently, TDP-43 was shown to be a major disease protein in the ubiquitinated inclusions characteristic of most cases of amyotrophic lateral sclerosis (ALS), tau-negative frontotemporal lobar degeneration (FTLD), and inclusion body myopathy. In these diseases, TDP-43 is redistributed from its predominantly nuclear location to ubiquitin-positive, cytoplasmic...

Freibaum, Brian D.; Chitta, Raghu; High, Anthony A.; Taylor, J. Paul

2010-01-01

78

Phosphorylated TDP-43 pathology and hippocampal sclerosis in progressive supranuclear palsy  

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TDP-43 is characteristically accumulated in TDP-43 proteinopathies such as frontotemporal lobar degeneration and motor neurone disease, but is also present in some tauopathies, including Alzheimer’s disease, argyrophilic grain disease, and corticobasal degeneration (CBD). However, several studies have suggested that cases of progressive supranuclear palsy (PSP) lack TDP-43 pathology. We have therefore examined limbic regions of the brain in 19 PSP cases, as well as in 12 CBD cases, using ph...

Yokota, Osamu; Davidson, Yvonne; Bigio, Eileen H.; Ishizu, Hideki; Terada, Seishi; Arai, Tetsuaki; Hasegawa, Masato; Akiyama, Haruhiko; Sikkink, Stephen; Pickering-brown, Stuart; Mann, David M. A.

2010-01-01

79

Disease Animal Models of TDP-43 Proteinopathy and Their Pre-Clinical Applications  

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Frontotemperal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS) are two common neurodegenerative diseases. TDP-43 is considered to be a major disease protein in FTLD/ALS, but it’s exact role in the pathogenesis and the effective treatments remains unknown. To address this question and to determine a potential treatment for FTLD/ALS, the disease animal models of TDP-43 proteinopathy have been established. TDP-43 proteinopathy is the histologic feature of FTLD/ALS and is asso...

Yu-Chih Liu; Po-Min Chiang; Kuen-Jer Tsai

2013-01-01

80

TDP-43 Is Directed to Stress Granules by Sorbitol, a Novel Physiological Osmotic and Oxidative Stressor ?  

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TDP-43, or TAR DNA-binding protein 43, is a pathological marker of a spectrum of neurodegenerative disorders, including amyotrophic lateral sclerosis and frontotemporal lobar degeneration with ubiquitin-positive inclusions. TDP-43 is an RNA/DNA-binding protein implicated in transcriptional and posttranscriptional regulation. Recent work also suggests that TDP-43 associates with cytoplasmic stress granules, which are transient structures that form in response to stress. In this study, we estab...

Dewey, Colleen M.; Cenik, Basar; Sephton, Chantelle F.; Dries, Daniel R.; Mayer, Paul; Good, Shannon K.; Johnson, Brett A.; Herz, Joachim; Yu, Gang

2011-01-01

 
 
 
 
81

Sustained Expression of TDP-43 and FUS in Motor Neurons in Rodent's Lifetime  

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TAR DNA-binding protein (TDP-43) and fused in sarcoma (FUS) are two highly conserved ribonucleoproteins. Pathogenic mutations of the TDP-43 or the FUS gene are all linked to amyotrophic lateral sclerosis (ALS) that is characterized by progressive degeneration of motor neurons. To better understand the correlation of ALS disease genes with the selectivity of chronic motor neuron degeneration, we examined the longitudinal expression of the TDP-43 and the FUS genes...

Cao Huang, Pedro Yuxing Xia

2010-01-01

82

Cytoplasmic accumulation of TDP-43 in circulating lymphomonocytes of ALS patients with and without TARDBP mutations  

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TDP-43, encoded by TARDBP, is a ubiquitously expressed, primarily nuclear protein. In recent years, TDP-43 has been identified as the major pathological protein in ALS due to its mislocalisation in the cytoplasm of motor neurons of patients with and without TARDBP mutations and expression in forms that do not match its predicted molecular weight. In this study, the TDP-43 profile was investigated using western immunoblot analysis in whole lysates, nuclei and cytoplasm of circulating lymphomon...

Giordana, Maria Teresa; Moglia, Cristina; Buccinna, Barbara; Ramondetti, Cristina; Piccinini, Marco; Chio, Adriano; Lupino, Elisa; Grifoni, Silvia; Marco, Giovanni; Lomartire, Annarosa; Calvo, Andrea; Rinaudo, Maria Teresa

2011-01-01

83

Tunicamycin produces TDP-43 cytoplasmic inclusions in cultured brain organotypic slices  

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The cellular distribution of TAR DNA binding protein (TDP-43) is disrupted in several neurodegenerative disorders, including frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U subtype) and amyotrophic lateral sclerosis (ALS). In these conditions, TDP-43 is found in neuronal cytoplasmic inclusions, with loss of the normal nuclear expression. The mechanisms leading to TDP-43 redistribution and its role in disease pathophysiology remain unknown. We describe an in vitro ...

Leggett, Cadman; Mcgehee, Daniel S.; Mastrianni, James; Yang, Wenbin; Bai, Tao; Brorson, James R.

2012-01-01

84

TDP-43 Potentiates Alpha-synuclein Toxicity to Dopaminergic Neurons in Transgenic Mice  

Directory of Open Access Journals (Sweden)

Full Text Available TDP-43 and ?-synuclein are two disease proteins involved in a wide range of neurodegenerative diseases. While TDP-43 proteinopathy is considered a pathologic hallmark of sporadic amyotrophic lateral sclerosis and frontotemporal lobe degeneration, ?-synuclein is a major component of Lewy body characteristic of Parkinson's disease. Intriguingly, TDP-43 proteinopathy also coexists with Lewy body and with synucleinopathy in certain disease conditions. Here we reported the effects of TDP-43 on ?-synuclein neurotoxicity in transgenic mice. Overexpression of mutant TDP-43 (M337V substitution in mice caused early death in transgenic founders, but overexpression of normal TDP-43 only induced a moderate loss of cortical neurons in the transgenic mice at advanced ages. Interestingly, concomitant overexpression of normal TDP-43 and mutant ?-synuclein caused a more severe loss of dopaminergic neurons in the double transgenic mice as compared to single-gene transgenic mice. TDP-43 potentiated ?-synuclein toxicity to dopaminergic neurons in living animals. Our finding provides in vivo evidence suggesting that disease proteins such as TDP-43 and ?-synuclein may play a synergistic role in disease induction in neurodegenerative diseases.

Tian Tian, Cao Huang, Jianbin Tong, Ming Yang, Hongxia Zhou, Xu-Gang Xia

2011-01-01

85

Axonal transport of TDP-43 mRNA granules is impaired by ALS-causing mutations.  

Science.gov (United States)

The RNA-binding protein TDP-43 regulates RNA metabolism at multiple levels, including transcription, RNA splicing, and mRNA stability. TDP-43 is a major component of the cytoplasmic inclusions characteristic of amyotrophic lateral sclerosis and some types of frontotemporal lobar degeneration. The importance of TDP-43 in disease is underscored by the fact that dominant missense mutations are sufficient to cause disease, although the role of TDP-43 in pathogenesis is unknown. Here we show that TDP-43 forms cytoplasmic mRNP granules that undergo bidirectional, microtubule-dependent transport in neurons in vitro and in vivo and facilitate delivery of target mRNA to distal neuronal compartments. TDP-43 mutations impair this mRNA transport function in vivo and in vitro, including in stem cell-derived motor neurons from ALS patients bearing any one of three different TDP-43 ALS-causing mutations. Thus, TDP-43 mutations that cause ALS lead to partial loss of a novel cytoplasmic function of TDP-43. PMID:24507191

Alami, Nael H; Smith, Rebecca B; Carrasco, Monica A; Williams, Luis A; Winborn, Christina S; Han, Steve S W; Kiskinis, Evangelos; Winborn, Brett; Freibaum, Brian D; Kanagaraj, Anderson; Clare, Alison J; Badders, Nisha M; Bilican, Bilada; Chaum, Edward; Chandran, Siddharthan; Shaw, Christopher E; Eggan, Kevin C; Maniatis, Tom; Taylor, J Paul

2014-02-01

86

Plasma phosphorylated-TDP-43 protein levels correlate with brain pathology in frontotemporal lobar degeneration  

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In the present study, we have correlated plasma TDP-43 levels, as measured by ELISA, with the presence of TDP-43 pathological changes in the brains of 28 patients with frontotemporal lobar degeneration (FTLD) (14 with FTLD-TDP and 14 with FTLD-tau) and 24 patients with pathologically confirmed AD (8 with, and 16 without, TDP-43 pathological changes). Western blotting revealed full-length TDP-43, including a phosphorylated form, and a phosphorylated C-terminal fragment, in all samples examined...

Foulds, Penelope G.; Davidson, Yvonne; Mishra, Manjari; Hobson, David J.; Humphreys, Kirsty M.; Taylor, Mark; Johnson, Nancy; Weintraub, Sandra; Akiyama, Haruhiko; Arai, Tetsuaki; Hasegawa, Masato; Bigio, Eileen H.; Benson, Fiona E.; Allsop, David; Mann, David M. A.

2009-01-01

87

Identification of Neuronal RNA Targets of TDP-43-containing Ribonucleoprotein Complexes*?  

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TAR DNA-binding protein 43 (TDP-43) is associated with a spectrum of neurodegenerative diseases. Although TDP-43 resembles heterogeneous nuclear ribonucleoproteins, its RNA targets and physiological protein partners remain unknown. Here we identify RNA targets of TDP-43 from cortical neurons by RNA immunoprecipitation followed by deep sequencing (RIP-seq). The canonical TDP-43 binding site (TG)n is 55.1-fold enriched, and moreover, a variant with adenine in the middle, (TG)nTA(TG)m, is highly...

Sephton, Chantelle F.; Cenik, Can; Kucukural, Alper; Dammer, Eric B.; Cenik, Basar; Han, Yuhong; Dewey, Colleen M.; Roth, Frederick P.; Herz, Joachim; Peng, Junmin; Moore, Melissa J.; Yu, Gang

2011-01-01

88

Tar DNA-binding protein-43 (TDP-43) regulates axon growth in vitro and in vivo.  

Science.gov (United States)

Intracellular inclusions of the TAR-DNA binding protein 43 (TDP-43) have been reported in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD-TDP). Rare mutations in TARDBP have been linked to both ALS and FTD-TDP suggesting that TDP-43 dysfunction is mechanistic in causing disease. TDP-43 is a predominantly nuclear protein with roles in regulating RNA transcription, splicing, stability and transport. In ALS, TDP-43 aberrantly accumulates in the cytoplasm of motor neurons where it forms aggregates. However it has until recently been unclear whether the toxic effects of TDP-43 involve recruitment to motor axons, and what effects this might have on axonal growth and integrity. Here we use chick embryonic motor neurons, in vivo and in vitro, to model the acute effects of TDP-43. We show that wild-type and two TDP-43 mutant proteins cause toxicity in chick embryonic motor neurons in vivo. Moreover, TDP-43 is increasingly mislocalised to axons over time in vivo, axon growth to peripheral targets is truncated, and expression of neurofilament-associated antigen is reduced relative to control motor neurons. In primary spinal motor neurons in vitro, a progressive translocation of TDP-43 to the cytoplasm occurs over time, similar to that observed in vivo. This coincides with the appearance of cytoplasmic aggregates, a reduction in the axonal length, and cellular toxicity, which was most striking for neurons expressing TDP-43 mutant forms. These observations suggest that the capacity of spinal motor neurons to produce and maintain an axon is compromised by dysregulation of TDP-43 and that the disruption of cytoskeletal integrity may play a role in the pathogenesis of ALS and FTD-TDP. PMID:24423647

Tripathi, Vineeta Bhasker; Baskaran, Pranetha; Shaw, Christopher E; Guthrie, Sarah

2014-05-01

89

Tar DNA-binding protein-43 (TDP-43) regulates axon growth in vitro and in vivo?  

Science.gov (United States)

Intracellular inclusions of the TAR-DNA binding protein 43 (TDP-43) have been reported in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD-TDP). Rare mutations in TARDBP have been linked to both ALS and FTD-TDP suggesting that TDP-43 dysfunction is mechanistic in causing disease. TDP-43 is a predominantly nuclear protein with roles in regulating RNA transcription, splicing, stability and transport. In ALS, TDP-43 aberrantly accumulates in the cytoplasm of motor neurons where it forms aggregates. However it has until recently been unclear whether the toxic effects of TDP-43 involve recruitment to motor axons, and what effects this might have on axonal growth and integrity. Here we use chick embryonic motor neurons, in vivo and in vitro, to model the acute effects of TDP-43. We show that wild-type and two TDP-43 mutant proteins cause toxicity in chick embryonic motor neurons in vivo. Moreover, TDP-43 is increasingly mislocalised to axons over time in vivo, axon growth to peripheral targets is truncated, and expression of neurofilament-associated antigen is reduced relative to control motor neurons. In primary spinal motor neurons in vitro, a progressive translocation of TDP-43 to the cytoplasm occurs over time, similar to that observed in vivo. This coincides with the appearance of cytoplasmic aggregates, a reduction in the axonal length, and cellular toxicity, which was most striking for neurons expressing TDP-43 mutant forms. These observations suggest that the capacity of spinal motor neurons to produce and maintain an axon is compromised by dysregulation of TDP-43 and that the disruption of cytoskeletal integrity may play a role in the pathogenesis of ALS and FTD-TDP.

Tripathi, Vineeta Bhasker; Baskaran, Pranetha; Shaw, Christopher E.; Guthrie, Sarah

2014-01-01

90

TDP-43 regulates the microprocessor complex activity during in vitro neuronal differentiation.  

Science.gov (United States)

TDP-43 (TAR DNA-binding protein 43) is an RNA-binding protein implicated in RNA metabolism at several levels. Even if ubiquitously expressed, it is considered as a neuronal activity-responsive factor and a major signature for neurological pathologies, making the comprehension of its activity in the nervous system a very challenging issue. TDP-43 has also been described as an accessory component of the Drosha-DGCR8 (DiGeorge syndrome critical region gene 8) microprocessor complex, which is crucially involved in basal and tissue-specific RNA processing events. In the present study, we exploited in vitro neuronal differentiation systems to investigate the TDP-43 demand for the microprocessor function, focusing on both its canonical microRNA biosynthetic activity and its alternative role as a post-transcriptional regulator of gene expression. Our findings reveal a novel role for TDP-43 as an essential factor that controls the stability of Drosha protein during neuronal differentiation, thus globally affecting the production of microRNAs. We also demonstrate that TDP-43 is required for the Drosha-mediated regulation of Neurogenin 2, a master gene orchestrating neurogenesis, whereas post-transcriptional control of Dgcr8, another Drosha target, resulted to be TDP-43-independent. These results implicate a previously uncovered contribution of TDP-43 in regulating the abundance and the substrate specificity of the microprocessor complex and provide new insights into TDP-43 as a key player in neuronal differentiation. PMID:24113842

Di Carlo, Valerio; Grossi, Elena; Laneve, Pietro; Morlando, Mariangela; Dini Modigliani, Stefano; Ballarino, Monica; Bozzoni, Irene; Caffarelli, Elisa

2013-12-01

91

Parkin reverses TDP-43-induced cell death and failure of amino acid homeostasis.  

Science.gov (United States)

The E3 ubiquitin ligase Parkin plays a central role in the pathogenesis of many neurodegenerative diseases. Parkin promotes specific ubiquitination and affects the localization of transactivation response DNA-binding protein 43 (TDP-43), which controls the translation of thousands of mRNAs. Here we tested the effects of lentiviral Parkin and TDP-43 expression on amino acid metabolism in the rat motor cortex using high frequency ¹³C NMR spectroscopy. TDP-43 expression increased glutamate levels, decreased the levels of other amino acids, including glutamine, aspartate, leucine and isoleucine, and impaired mitochondrial tricarboxylic acid cycle. TDP-43 induced lactate accumulation and altered the balance between excitatory (glutamate) and inhibitory (GABA) neurotransmitters. Parkin restored amino acid levels, neurotransmitter balance and tricarboxylic acid cycle metabolism, rescuing neurons from TDP-43-induced apoptotic death. Furthermore, TDP-43 expression led to an increase in 4E-BP levels, perhaps altering translational control and deregulating amino acid synthesis; while Parkin reversed the effects of TDP-43 on the 4E-BP signaling pathway. Taken together, these data suggest that Parkin may affect TDP-43 localization and mitigate its effects on 4E-BP signaling and loss of amino acid homeostasis. PMID:24298989

Hebron, Michaeline; Chen, Wenqiang; Miessau, Matthew J; Lonskaya, Irina; Moussa, Charbel E-H

2014-04-01

92

Inhibition of RNA lariat debranching enzyme suppresses TDP-43 toxicity in ALS disease models.  

Science.gov (United States)

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease primarily affecting motor neurons. Mutations in the gene encoding TDP-43 cause some forms of the disease, and cytoplasmic TDP-43 aggregates accumulate in degenerating neurons of most individuals with ALS. Thus, strategies aimed at targeting the toxicity of cytoplasmic TDP-43 aggregates may be effective. Here, we report results from two genome-wide loss-of-function TDP-43 toxicity suppressor screens in yeast. The strongest suppressor of TDP-43 toxicity was deletion of DBR1, which encodes an RNA lariat debranching enzyme. We show that, in the absence of Dbr1 enzymatic activity, intronic lariats accumulate in the cytoplasm and likely act as decoys to sequester TDP-43, preventing it from interfering with essential cellular RNAs and RNA-binding proteins. Knockdown of Dbr1 in a human neuronal cell line or in primary rat neurons is also sufficient to rescue TDP-43 toxicity. Our findings provide insight into TDP-43-mediated cytotoxicity and suggest that decreasing Dbr1 activity could be a potential therapeutic approach for ALS. PMID:23104007

Armakola, Maria; Higgins, Matthew J; Figley, Matthew D; Barmada, Sami J; Scarborough, Emily A; Diaz, Zamia; Fang, Xiaodong; Shorter, James; Krogan, Nevan J; Finkbeiner, Steven; Farese, Robert V; Gitler, Aaron D

2012-12-01

93

The N-terminus of TDP-43 promotes its oligomerization and enhances DNA binding affinity  

Energy Technology Data Exchange (ETDEWEB)

Highlights: Black-Right-Pointing-Pointer The N-terminus of TDP-43 contains an independently folded structural domain (NTD). Black-Right-Pointing-Pointer The structural domains of TDP-43 are arranged in a beads-on-a-string fashion. Black-Right-Pointing-Pointer The NTD promotes TDP-43 oligomerization in a concentration-dependent manner. Black-Right-Pointing-Pointer The NTD may assist nucleic acid-binding activity of TDP-43. -- Abstract: TDP-43 is a DNA/RNA-binding protein associated with different neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD-U). Here, the structural and physical properties of the N-terminus on TDP-43 have been carefully characterized through a combination of nuclear magnetic resonance (NMR), circular dichroism (CD) and fluorescence anisotropy studies. We demonstrate for the first time the importance of the N-terminus in promoting TDP-43 oligomerization and enhancing its DNA-binding affinity. An unidentified structural domain in the N-terminus is also disclosed. Our findings provide insights into the N-terminal domain function of TDP-43.

Chang, Chung-ke [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Wu, Tzong-Huah [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China); Chemical Biology and Molecular Biophysics Program, Taiwan International Graduate Program, Institute of Biochemistry, Academia Sinica, Taipei 115, Taiwan (China); Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 300, Taiwan (China); Wu, Chu-Ya [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China); Graduate Institute of Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan (China); Chiang, Ming-hui; Toh, Elsie Khai-Woon [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Hsu, Yin-Chih; Lin, Ku-Feng [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China); Liao, Yu-heng [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Huang, Tai-huang, E-mail: bmthh@gate.sinica.edu.tw [Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan (China); Department of Physics, National Taiwan Normal University, Taipei 106, Taiwan (China); Huang, Joseph Jen-Tse, E-mail: jthuang@chem.sinica.edu.tw [Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan (China)

2012-08-24

94

C-Jun N-terminal kinase controls TDP-43 accumulation in stress granules induced by oxidative stress  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background TDP-43 proteinopathies are characterized by loss of nuclear TDP-43 expression and formation of C-terminal TDP-43 fragmentation and accumulation in the cytoplasm. Recent studies have shown that TDP-43 can accumulate in RNA stress granules (SGs in response to cell stresses and this could be associated with subsequent formation of TDP-43 ubiquinated protein aggregates. However, the initial mechanisms controlling endogenous TDP-43 accumulation in SGs during chronic disease are not understood. In this study we investigated the mechanism of TDP-43 processing and accumulation in SGs in SH-SY5Y neuronal-like cells exposed to chronic oxidative stress. Cell cultures were treated overnight with the mitochondrial inhibitor paraquat and examined for TDP-43 and SG processing. Results We found that mild stress induced by paraquat led to formation of TDP-43 and HuR-positive SGs, a proportion of which were ubiquitinated. The co-localization of TDP-43 with SGs could be fully prevented by inhibition of c-Jun N-terminal kinase (JNK. JNK inhibition did not prevent formation of HuR-positive SGs and did not prevent diffuse TDP-43 accumulation in the cytosol. In contrast, ERK or p38 inhibition prevented formation of both TDP-43 and HuR-positive SGs. JNK inhibition also inhibited TDP-43 SG localization in cells acutely treated with sodium arsenite and reduced the number of aggregates per cell in cultures transfected with C-terminal TDP-43 162-414 and 219-414 constructs. Conclusions Our studies are the first to demonstrate a critical role for kinase control of TDP-43 accumulation in SGs and may have important implications for development of treatments for FTD and ALS, targeting cell signal pathway control of TDP-43 aggregation.

Masters Colin L

2011-08-01

95

Multiplex SILAC Analysis of a Cellular TDP-43 Proteinopathy Model Reveals Protein Inclusions Associated with SUMOylation and Diverse Polyubiquitin Chains*  

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Transactive response (TAR) DNA-binding protein 43 (TDP-43) is a major protein component within ubiquitin-positive inclusions of frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Although TDP-43 is a nuclear DNA/RNA-binding protein, in pathological conditions, TDP-43 has been reported to redistribute to the cytoplasm where it is cleaved and forms insoluble, ubiquitinated, and phosphorylated inclusions. Here we present a cellular model in which full-length human TDP-43 or a s...

Seyfried, Nicholas T.; Gozal, Yair M.; Dammer, Eric B.; Xia, Qiangwei; Duong, Duc M.; Cheng, Dongmei; Lah, James J.; Levey, Allan I.; Peng, Junmin

2010-01-01

96

TDP-43 Is Intrinsically Aggregation-prone, and Amyotrophic Lateral Sclerosis-linked Mutations Accelerate Aggregation and Increase Toxicity*  

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Non-amyloid, ubiquitinated cytoplasmic inclusions containing TDP-43 and its C-terminal fragments are pathological hallmarks of amyotrophic lateral sclerosis (ALS), a fatal motor neuron disorder, and frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U). Importantly, TDP-43 mutations are linked to sporadic and non-SOD1 familial ALS. However, TDP-43 is not the only protein in disease-associated inclusions, and whether TDP-43 misfolds or is merely sequestered by other agg...

Johnson, Brian S.; Snead, David; Lee, Jonathan J.; Mccaffery, J. Michael; Shorter, James; Gitler, Aaron D.

2009-01-01

97

The dual functions of the extreme N-terminus of TDP-43 in regulating its biological activity and inclusion formation  

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TAR DNA-binding protein-43 (TDP-43) is the principal component of ubiquitinated inclusions in amyotrophic lateral sclerosis (ALS) and the most common pathological subtype of frontotemporal dementia—frontotemporal lobar degeneration with TDP-43-positive inclusions (FTLD-TDP). To date, the C-terminus of TDP-43, which is aggregation-prone and contains almost all ALS-associated mutations, has garnered much attention while the functions of the N-terminus of TDP-43 remain largely unknown. To brid...

Zhang, Yong-jie; Caulfield, Thomas; Xu, Ya-fei; Gendron, Tania F.; Hubbard, Jaime; Stetler, Caroline; Sasaguri, Hiroki; Whitelaw, Ena C.; Cai, Shuyi; Lee, Wing Cheung; Petrucelli, Leonard

2013-01-01

98

Reduced cellular Ca(2+) availability enhances TDP-43 cleavage by apoptotic caspases.  

Science.gov (United States)

Accumulation of transactive response DNA binding protein (TDP-43) fragments in motor neurons is a post mortem hallmark of different neurodegenerative diseases. TDP-43 fragments are the products of the apoptotic caspases-3 and -7. Either excessive or insufficient cellular Ca(2+) availability is associated with activation of apoptotic caspases. However, as far as we know, it is not described whether activation of caspases, due to restricted intracellular Ca(2+), affects TDP-43 cleavage. Here we show that in various cell lineages with restricted Ca(2+) availability, TDP-43 is initially cleaved by caspases-3 and -7 and then, also by caspases-6 and -8 once activated by caspase-3. Furthermore, we disclose the existence of a TDP-43 caspase-mediated fragment of 15kDa, in addition to the well-known fragments of 35 and 25kDa. Interestingly, with respect to the other two fragments this novel fragment is the major product of caspase activity on murine TDP-43 whereas in human cell lines the opposite occurs. This outcome should be considered when murine models are used to investigate TDP-43 proteinopathies. PMID:24440855

De Marco, Giovanni; Lomartire, Annarosa; Mandili, Giorgia; Lupino, Elisa; Buccinnà, Barbara; Ramondetti, Cristina; Moglia, Cristina; Novelli, Francesco; Piccinini, Marco; Mostert, Michael; Rinaudo, Maria Teresa; Chiò, Adriano; Calvo, Andrea

2014-04-01

99

TDP-43 knockdown impairs neurite outgrowth dependent on its target histone deacetylase 6  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Trans-activation response element (TAR DNA binding protein of 43kDa (TDP-43 is causally related to the neurodegenerative diseases frontotemporal dementia and amyotrophic lateral sclerosis being the hallmark protein in the disease-characteristic neuropathological lesions and via genetic linkage. Histone deacetylase 6 (HDAC6 is an established target of the RNA-binding protein TDP-43. HDAC6 is an unusual cytosolic deacetylase enzyme, central for a variety of pivotal cellular functions including aggregating protein turnover, microtubular dynamics and filopodia formation. All these functions are important in the context of neurodegenerative proteinopathies involving TDP-43. We have previously shown in a human embryonic kidney cell line that TDP-43 knockdown significantly impairs the removal of a toxic, aggregating polyQ ataxin-3 fusion protein in an HDAC6-dependent manner. Here we investigated the influence of TDP-43 and its target HDAC6 on neurite outgrowth. Results Human neuroblastoma SH-SY5Y cells with stably silenced TDP-43 showed a significant reduction of neurite outgrowth induced by retinoic acid and brain-derived neurotrophic factor. Re-transfection with TDP-43 as well as HDAC6 rescued retinoic acid-induced neurite outgrowth. In addition, we show that silencing of HDAC6 alone is sufficient to reduce neurite outgrowth of in vitro differentiated SH-SY5Y cells. Conclusions TDP-43 deficiency leads to impairment of neurite growth in an HDAC6-dependent manner, thereby contributing to neurodegenerative events in TDP-43 diseases.

Weber Stephanie S

2011-08-01

100

Amyotrophic lateral sclerosis and frontotemporal lobar degeneration: A spectrum of TDP-43 proteinopathies  

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It is now established that pathological transactive response DNA-binding protein with a Mr of 43 kD (TDP-43) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis is the major disease protein in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) with ubiquitin-positive inclusions (now known as FTLD-TDP). In fact, the discovery of pathological TDP-43 solidified the idea that these disorders are multi-system diseases and this led to the concept of a TDP-43 p...

Geser, Felix; Lee, Virginia M. -y; Trojanowski, John Q.

2010-01-01

 
 
 
 
101

Understanding the role of TDP-43 and FUS/TLS in ALS and beyond  

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Dominant mutation in two DNA/RNA binding proteins, TDP-43 and FUS/TLS, are causes of inherited Amyotrophic Lateral Sclerosis (ALS). TDP-43 and FUS/TLS have striking structural and functional similarities, implicating alterations in RNA processing as central in ALS. TDP-43 has binding sites within a third of all mouse and human mRNAs in brain and this binding influences the levels and splicing patterns of at least 20% of those mRNAs. Disease modeling in rodents of the first known cause of inhe...

2011-01-01

102

Differential roles of the ubiquitin proteasome system and autophagy in the clearance of soluble and aggregated TDP-43 species  

Science.gov (United States)

ABSTRACT TAR DNA-binding protein (TDP-43, also known as TARDBP) is the major pathological protein in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Large TDP-43 aggregates that are decorated with degradation adaptor proteins are seen in the cytoplasm of remaining neurons in ALS and FTD patients post mortem. TDP-43 accumulation and ALS-linked mutations within degradation pathways implicate failed TDP-43 clearance as a primary disease mechanism. Here, we report the differing roles of the ubiquitin proteasome system (UPS) and autophagy in the clearance of TDP-43. We have investigated the effects of inhibitors of the UPS and autophagy on the degradation, localisation and mobility of soluble and insoluble TDP-43. We find that soluble TDP-43 is degraded primarily by the UPS, whereas the clearance of aggregated TDP-43 requires autophagy. Cellular macroaggregates, which recapitulate many of the pathological features of the aggregates in patients, are reversible when both the UPS and autophagy are functional. Their clearance involves the autophagic removal of oligomeric TDP-43. We speculate that, in addition to an age-related decline in pathway activity, a second hit in either the UPS or the autophagy pathway drives the accumulation of TDP-43 in ALS and FTD. Therapies for clearing excess TDP-43 should therefore target a combination of these pathways.

Scotter, Emma L.; Vance, Caroline; Nishimura, Agnes L.; Lee, Youn-Bok; Chen, Han-Jou; Urwin, Hazel; Sardone, Valentina; Mitchell, Jacqueline C.; Rogelj, Boris; Rubinsztein, David C.; Shaw, Christopher E.

2014-01-01

103

Differential roles of the ubiquitin proteasome system and autophagy in the clearance of soluble and aggregated TDP-43 species.  

Science.gov (United States)

TAR DNA-binding protein (TDP-43, also known as TARDBP) is the major pathological protein in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Large TDP-43 aggregates that are decorated with degradation adaptor proteins are seen in the cytoplasm of remaining neurons in ALS and FTD patients post mortem. TDP-43 accumulation and ALS-linked mutations within degradation pathways implicate failed TDP-43 clearance as a primary disease mechanism. Here, we report the differing roles of the ubiquitin proteasome system (UPS) and autophagy in the clearance of TDP-43. We have investigated the effects of inhibitors of the UPS and autophagy on the degradation, localisation and mobility of soluble and insoluble TDP-43. We find that soluble TDP-43 is degraded primarily by the UPS, whereas the clearance of aggregated TDP-43 requires autophagy. Cellular macroaggregates, which recapitulate many of the pathological features of the aggregates in patients, are reversible when both the UPS and autophagy are functional. Their clearance involves the autophagic removal of oligomeric TDP-43. We speculate that, in addition to an age-related decline in pathway activity, a second hit in either the UPS or the autophagy pathway drives the accumulation of TDP-43 in ALS and FTD. Therapies for clearing excess TDP-43 should therefore target a combination of these pathways. PMID:24424030

Scotter, Emma L; Vance, Caroline; Nishimura, Agnes L; Lee, Youn-Bok; Chen, Han-Jou; Urwin, Hazel; Sardone, Valentina; Mitchell, Jacqueline C; Rogelj, Boris; Rubinsztein, David C; Shaw, Christopher E

2014-03-15

104

Differential diagnosis of amyotrophic lateral sclerosis from Guillain-Barré syndrome by quantitative determination of TDP-43 in cerebrospinal fluid.  

Science.gov (United States)

The aim of this study was to investigate whether an increased level of TAR DNA-binding protein 43 (TDP-43) in the cerebrospinal fluid (CSF) could be a biomarker for amyotrophic lateral sclerosis (ALS) and facilitate differential diagnosis of ALS from peripheral motor neuropathy. TDP-43 is the major constituent of neuronal and glial inclusions that neuropathologically characterize both ALS and tau-negative frontotemporal lobar degeneration. Recent discoveries of various missense mutations in the TDP-43 gene in familial ALS indicate a pivotal role of the aberrant accumulation of TDP-43 in neurodegeneration. Increased TDP-43 in the CSF could be a hallmark of ALS and other TDP-43 proteinopathy. Sandwich enzyme-linked immunosorbent assay (ELISA) was established to measure the concentration of TDP-43 in biological fluids. Culture supernatants of cells transfected with various TDP-43 constructs were used to confirm that the ELISA detected TDP-43. TDP-43 in the culture supernatant of TDP-43 transfected cells was detected by immunoprecipitation with subsequent immunoblotting and concentrations were successfully measured by sandwich ELISA. We then measured TDP-43 concentrations in the CSF of patients with ALS and Guillain-Barré syndrome (GBS). TDP-43 concentrations in CSF were significantly higher in ALS than in GBS (p = 0.016). The sensitivity of the diagnostic test was 71.4% and the specificity was 84.6%. Quantitative determination of TDP-43 concentrations in the CSF by sandwich ELISA is a potential laboratory test for differentiating ALS from peripheral motor neuropathies such as GBS. PMID:24066851

Hosokawa, Masato; Arai, Tetsuaki; Yamashita, Makiko; Tsuji, Hiroshi; Nonaka, Takashi; Masuda-Suzukake, Masami; Tamaoka, Akira; Hasegawa, Masato; Akiyama, Haruhiko

2014-05-01

105

Robust cytoplasmic accumulation of phosphorylated TDP-43 in transgenic models of tauopathy  

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Frontotemporal lobar degeneration (FTLD) has been subdivided based on the main pathology found in the brains of affected individuals. When the primary pathology is aggregated, hyperphosphorylated tau, the pathological diagnosis is FTLD-tau. When the primary pathology is cytoplasmic and/or nuclear aggregates of phosphorylated TAR-DNA-binding protein (TDP-43), the pathological diagnosis is FTLD-TDP. Notably, TDP-43 pathology can also occur in conjunction with a number of neurodegenerative disor...

Clippinger, Amy K.; D’alton, Simon; Lin, Wen-lang; Gendron, Tania F.; Howard, John; Borchelt, David R.; Cannon, Ashley; Carlomagno, Yari; Chakrabarty, Paramita; Cook, Casey; Golde, Todd E.; Levites, Yona; Ranum, Laura; Schultheis, Patrick J.; Xu, Guilian

2013-01-01

106

TDP-43 knockdown impairs neurite outgrowth dependent on its target histone deacetylase 6  

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Abstract Background Trans-activation response element (TAR) DNA binding protein of 43kDa (TDP-43) is causally related to the neurodegenerative diseases frontotemporal dementia and amyotrophic lateral sclerosis being the hallmark protein in the disease-characteristic neuropathological lesions and via genetic linkage. Histone deacetylase 6 (HDAC6) is an established target of the RNA-binding protein TDP-43. HDAC6 is an unusual cytosolic deacetylase enzyme, central for a variety ...

Fiesel Fabienne C; Schurr Christine; Weber Stephanie S; Kahle Philipp J

2011-01-01

107

Different 8-hydroxyquinolines protect models of TDP-43 protein, ?-synuclein, and polyglutamine proteotoxicity through distinct mechanisms.  

Science.gov (United States)

No current therapies target the underlying cellular pathologies of age-related neurodegenerative diseases. Model organisms provide a platform for discovering compounds that protect against the toxic, misfolded proteins that initiate these diseases. One such protein, TDP-43, is implicated in multiple neurodegenerative diseases, including amyotrophic lateral sclerosis and frontotemporal lobar degeneration. In yeast, TDP-43 expression is toxic, and genetic modifiers first discovered in yeast have proven to modulate TDP-43 toxicity in both neurons and humans. Here, we describe a phenotypic screen for small molecules that reverse TDP-43 toxicity in yeast. One group of hit compounds was 8-hydroxyquinolines (8-OHQ), a class of clinically relevant bioactive metal chelators related to clioquinol. Surprisingly, in otherwise wild-type yeast cells, different 8-OHQs had selectivity for rescuing the distinct toxicities caused by the expression of TDP-43, ?-synuclein, or polyglutamine proteins. In fact, each 8-OHQ synergized with the other, clearly establishing that they function in different ways. Comparative growth and molecular analyses also revealed that 8-OHQs have distinct metal chelation and ionophore activities. The diverse bioactivity of 8-OHQs indicates that altering different aspects of metal homeostasis and/or metalloprotein activity elicits distinct protective mechanisms against several neurotoxic proteins. Indeed, phase II clinical trials of an 8-OHQ has produced encouraging results in modifying Alzheimer disease. Our unbiased identification of 8-OHQs in a yeast TDP-43 toxicity model suggests that tailoring 8-OHQ activity to a particular neurodegenerative disease may be a viable therapeutic strategy. PMID:22147697

Tardiff, Daniel F; Tucci, Michelle L; Caldwell, Kim A; Caldwell, Guy A; Lindquist, Susan

2012-02-01

108

Understanding the role of TDP-43 and FUS/TLS in ALS and beyond.  

Science.gov (United States)

Dominant mutations in two DNA/RNA binding proteins, TDP-43 and FUS/TLS, are causes of inherited Amyotrophic Lateral Sclerosis (ALS). TDP-43 and FUS/TLS have striking structural and functional similarities, implicating alterations in RNA processing as central in ALS. TDP-43 has binding sites within a third of all mouse and human mRNAs in brain and this binding influences the levels and splicing patterns of at least 20% of those mRNAs. Disease modeling in rodents of the first known cause of inherited ALS-mutation in the ubiquitously expressed superoxide dismutase (SOD1)-has yielded non-cell autonomous fatal motor neuron disease caused by one or more toxic properties acquired by the mutant proteins. In contrast, initial disease modeling for TDP-43 and FUS/TLS has produced highly varied phenotypes. It remains unsettled whether TDP-43 and FUS/TLS mutants provoke disease from a loss of function or gain of toxicity or both. TDP-43 or FUS/TLS misaccumulation seems central not just to ALS (where it is found in almost all instances of disease), but more broadly in neurodegenerative disease, including frontal temporal lobular dementia (FTLD-U) and many examples of Alzheimer's or Huntington's disease. PMID:21813273

Da Cruz, Sandrine; Cleveland, Don W

2011-12-01

109

TDP-43 neurotoxicity and protein aggregation modulated by heat shock factor and insulin/IGF-1 signaling  

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TAR DNA-binding protein 43 (TDP-43) plays a key role in the neurodegenerative diseases including amyotrophic lateral sclerosis and frontotemporal lobar degeneration. The nature of the TDP-43-mediated neurotoxicity associated with these diseases is not yet understood. Here, we have established transgenic Caenorhabditis elegans models that express human TDP-43 variants in the nervous system, including the full-length wild-type (WT) and mutant proteins and a pathologic C-terminal fragment. The C...

Zhang, Tao; Mullane, Patrick C.; Periz, Goran; Wang, Jiou

2011-01-01

110

Genetic strategies to study TDP-43 in rodents and to develop preclinical therapeutics for amyotrophic lateral sclerosis  

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The neuropathological hallmark of the majority of amyotrophic lateral sclerosis (ALS) and a class of frontotemporal lobar degeneration is ubiquitinated cytoplasmic aggregates composed of transactive response DNA binding protein 43 kDa (TDP-43). Genetic manipulation of TDP-43 in animal models has been used to study the protein's role in pathogenesis. Transgenic rodents for TDP-43 have recapitulated key aspects of ALS such as paralysis, loss of spinal motor neurons and muscle atrophy. Viral vec...

Wang, David B.; Gitcho, Michael A.; Kraemer, Brian C.; Klein, Ronald L.

2011-01-01

111

Dysregulation of the ALS-associated gene TDP-43 leads to neuronal death and degeneration in mice  

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Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are characterized by cytoplasmic protein aggregates in the brain and spinal cord that include TAR-DNA binding protein 43 (TDP-43). TDP-43 is normally localized in the nucleus with roles in the regulation of gene expression, and pathological cytoplasmic aggregates are associated with depletion of nuclear protein. Here, we generated transgenic mice expressing human TDP-43 with a defective nuclear localization signa...

Igaz, Lionel M.; Kwong, Linda K.; Lee, Edward B.; Chen-plotkin, Alice; Swanson, Eric; Unger, Travis; Malunda, Joe; Xu, Yan; Winton, Matthew J.; Trojanowski, John Q.; Lee, Virginia M. -y

2011-01-01

112

TDP-43 inclusion bodies formed in bacteria are structurally amorphous, non-amyloid and inherently toxic to neuroblastoma cells.  

Science.gov (United States)

Accumulation of ubiquitin-positive, tau- and ?-synuclein-negative intracellular inclusions of TDP-43 in the central nervous system represents the major hallmark correlated to amyotrophic lateral sclerosis and frontotemporal lobar degeneration with ubiquitin-positive inclusions. Such inclusions have variably been described as amorphous aggregates or more structured deposits having an amyloid structure. Following the observations that bacterial inclusion bodies generally consist of amyloid aggregates, we have overexpressed full-length TDP-43 and C-terminal TDP-43 in E. coli, purified the resulting full-length and C-terminal TDP-43 containing inclusion bodies (FL and Ct TDP-43 IBs) and subjected them to biophysical analyses to assess their structure/morphology. We show that both FL and Ct TDP-43 aggregates contained in the bacterial IBs do not bind amyloid dyes such as thioflavin T and Congo red, possess a disordered secondary structure, as inferred using circular dichroism and infrared spectroscopies, and are susceptible to proteinase K digestion, thus possessing none of the hallmarks for amyloid. Moreover, atomic force microscopy revealed an irregular structure for both types of TDP-43 IBs and confirmed the absence of amyloid-like species after proteinase K treatment. Cell biology experiments showed that FL TDP-43 IBs were able to impair the viability of cultured neuroblastoma cells when added to their extracellular medium and, more markedly, when transfected into their cytosol, where they are at least in part ubiquitinated and phosphorylated. These data reveal an inherently high propensity of TDP-43 to form amorphous aggregates, which possess, however, an inherently high ability to cause cell dysfunction. This indicates that a gain of toxic function caused by TDP-43 deposits is effective in TDP-43 pathologies, in addition to possible loss of function mechanisms originating from the cellular mistrafficking of the protein. PMID:24497973

Capitini, Claudia; Conti, Simona; Perni, Michele; Guidi, Francesca; Cascella, Roberta; De Poli, Angela; Penco, Amanda; Relini, Annalisa; Cecchi, Cristina; Chiti, Fabrizio

2014-01-01

113

TDP-43 Inclusion Bodies Formed in Bacteria Are Structurally Amorphous, Non-Amyloid and Inherently Toxic to Neuroblastoma Cells  

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Accumulation of ubiquitin-positive, tau- and ?-synuclein-negative intracellular inclusions of TDP-43 in the central nervous system represents the major hallmark correlated to amyotrophic lateral sclerosis and frontotemporal lobar degeneration with ubiquitin-positive inclusions. Such inclusions have variably been described as amorphous aggregates or more structured deposits having an amyloid structure. Following the observations that bacterial inclusion bodies generally consist of amyloid aggregates, we have overexpressed full-length TDP-43 and C-terminal TDP-43 in E. coli, purified the resulting full-length and C-terminal TDP-43 containing inclusion bodies (FL and Ct TDP-43 IBs) and subjected them to biophysical analyses to assess their structure/morphology. We show that both FL and Ct TDP-43 aggregates contained in the bacterial IBs do not bind amyloid dyes such as thioflavin T and Congo red, possess a disordered secondary structure, as inferred using circular dichroism and infrared spectroscopies, and are susceptible to proteinase K digestion, thus possessing none of the hallmarks for amyloid. Moreover, atomic force microscopy revealed an irregular structure for both types of TDP-43 IBs and confirmed the absence of amyloid-like species after proteinase K treatment. Cell biology experiments showed that FL TDP-43 IBs were able to impair the viability of cultured neuroblastoma cells when added to their extracellular medium and, more markedly, when transfected into their cytosol, where they are at least in part ubiquitinated and phosphorylated. These data reveal an inherently high propensity of TDP-43 to form amorphous aggregates, which possess, however, an inherently high ability to cause cell dysfunction. This indicates that a gain of toxic function caused by TDP-43 deposits is effective in TDP-43 pathologies, in addition to possible loss of function mechanisms originating from the cellular mistrafficking of the protein.

Capitini, Claudia; Conti, Simona; Perni, Michele; Guidi, Francesca; Cascella, Roberta; De Poli, Angela; Penco, Amanda; Relini, Annalisa; Cecchi, Cristina; Chiti, Fabrizio

2014-01-01

114

Endogenous TDP-43, but not FUS, contributes to stress granule assembly via G3BP  

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Full Text Available Abstract Amyotrophic lateral sclerosis (ALS is a fatal neurodegenerative disease characterized by the selective loss of upper and lower motor neurons, a cell type that is intrinsically more vulnerable than other cell types to exogenous stress. The interplay between genetic susceptibility and environmental exposures to toxins has long been thought to be relevant to ALS. One cellular mechanism to overcome stress is the formation of small dense cytoplasmic domains called stress granules (SG which contain translationally arrested mRNAs. TDP-43 (encoded by TARDBP is an ALS-causative gene that we have previously implicated in the regulation of the core stress granule proteins G3BP and TIA-1. TIA-1 and G3BP localize to SG under nearly all stress conditions and are considered essential to SG formation. Here, we report that TDP-43 is required for proper SG dynamics, especially SG assembly as marked by the secondary aggregation of TIA-1. We also show that SG assembly, but not initiation, requires G3BP. Furthermore, G3BP can rescue defective SG assembly in cells depleted of endogenous TDP-43. We also demonstrate that endogenous TDP-43 and FUS do not have overlapping functions in this cellular process as SG initiation and assembly occur normally in the absence of FUS. Lastly, we observe that SG assembly is a contributing factor in the survival of neuronal-like cells responding to acute oxidative stress. These data raise the possibility that disruptions of normal stress granule dynamics by loss of nuclear TDP-43 function may contribute to neuronal vulnerability in ALS.

Aulas Anaïs

2012-10-01

115

Coaggregation of RNA-Binding Proteins in a Model of TDP-43 Proteinopathy with Selective RGG Motif Methylation and a Role for RRM1 Ubiquitination  

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TAR DNA-binding protein 43 (TDP-43) is a major component within ubiquitin-positive inclusions of a number of neurodegenerative diseases that increasingly are considered as TDP-43 proteinopathies. Identities of other inclusion proteins associated with TDP-43 aggregation remain poorly defined. In this study, we identify and quantitate 35 co-aggregating proteins in the detergent-resistant fraction of HEK-293 cells in which TDP-43 or a particularly aggregate prone variant, TDP-S6, were enriched f...

Dammer, Eric B.; Fallini, Claudia; Gozal, Yair M.; Duong, Duc M.; Rossoll, Wilfried; Xu, Ping; Lah, James J.; Levey, Allan I.; Peng, Junmin; Bassell, Gary J.; Seyfried, Nicholas T.

2012-01-01

116

The ALS-Linked Gene TDP-43 Regulates \\(IFN\\beta\\) Expression through a Novel Mechanism of 3' UTR-Mediated Promoter cis-Regulation  

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The TAR DNA-binding protein (TDP-43) is a heterogeneous nuclear ribonucleprotein that is involved in multiple stages of RNA processing. Mutations in the TDP-43 gene and mislocalization of TDP-43 protein have been implicated in a growing number of neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Here, we show that TDP-43 negatively regulates innate immune gene expression in response to RNA virus sensing. Perturbation of TDP...

Deering, Raquel Payzant

2012-01-01

117

On the development of markers for pathological TDP-43 in amyotrophic lateral sclerosis with and without dementia.  

LENUS (Irish Health Repository)

Pathological 43-kDa transactive response sequence DNA-binding protein (TDP-43) has been recognized as the major disease protein in amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration with ubiquitin positive, tau and ?-synuclein negative inclusions (FTLD-U) and the transitional forms between these multisystem conditions. In order to develop TDP-43 into a successful ALS biomarker, the natural history of TDP-43 pathology needs to be characterized and the underlying pathophysiology established. Here we propose a spatial and temporal "two-axes" model of central nervous system vulnerability for TDP-43 linked degeneration and review recent studies on potential biomarkers related to pathological TDP-43 in the cerebrospinal fluid (CSF), blood, and skeletal muscle. The model includes the following two arms: Firstly, a "motor neuron disease" or "spinal cord\\/brainstem to motor cortex" axis (with degeneration possibly ascending from the lower motor neurons to the upper motor neurons); and secondly, a "dementia" or "corticoid\\/allocortex to neocortex" axis (with a probable spread of TDP-43 linked degeneration from the mediotemporal lobe to wider mesocortical and neocortical brain areas). At the cellular level, there is a gradual disappearance of normal TDP-43 in the nucleus in combination with the formation of pathological aggregates in the cell body and cellular processes, which can also be used to identify the stage of the disease process. Moreover, TDP-43 lesions in subpial\\/subependymal or perivascular localizations have been noted, and this might account for increased CSF and blood TDP-43 levels through mechanisms that remain to be elucidated.

Geser, F

2011-12-01

118

Autoregulation of TDP-43 mRNA levels involves interplay between transcription, splicing, and alternative polyA site selection  

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Implicated in many neuropathologies, TDP-43 autoregulates its expression via a negative feedback loop. Baralle and colleagues now elucidate the interesting mechanism underlying this autoregulation. TDP-43 binding to its pre-mRNA 3? UTR blocks recognition of the main pA site and also causes Pol II pausing downstream from its binding site, leading to excision of the main pA site via splicing of a 3? UTR intron. This dual attack rapidly reduces levels of correctly processed TDP-43 mRNA.

Avendan?o-va?zquez, S. Ere?ndira; Dhir, Ashish; Bembich, Sara; Buratti, Emanuele; Proudfoot, Nicholas; Baralle, Francisco E.

2012-01-01

119

FUS pathology defines the majority of tau- and TDP-43-negative frontotemporal lobar degeneration  

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Through an international consortium, we have collected 37 tau- and TAR DNA-binding protein 43 (TDP-43)-negative frontotemporal lobar degeneration (FTLD) cases, and present here the first comprehensive analysis of these cases in terms of neuropathology, genetics, demographics and clinical data. 92% (34/37) had fused in sarcoma (FUS) protein pathology, indicating that FTLD-FUS is an important FTLD subtype. This FTLD-FUS collection specifically focussed on aFTLD-U cases, one of three recently de...

Urwin, Hazel; Josephs, Keith A.; Rohrer, Jonathan D.; Mackenzie, Ian R.; Neumann, Manuela; Authier, Astrid; Seelaar, Harro; Swieten, John C.; Brown, Jeremy M.; Johannsen, Peter; Nielsen, Jorgen E.; Holm, Ida E.; Dickson, Dennis W.; Rademakers, Rosa; Graff-radford, Neill R.

2010-01-01

120

Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation.  

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TDP-43 is a nuclear protein involved in many aspects of RNA metabolism. To ensure cellular viability, its expression levels within cells must be tightly regulated. We have previously demonstrated that TDP-43 autoregulation occurs through the activation of a normally silent intron in its 3'-UTR sequence that results in the use of alternative polyadenylation sites. In this work, we analyse which is the dominant event in autoregulation: the recognition of the splice sites of 3'-UTR intron 7 or the intrinsic quality of the alternative polyadenylation sites. A panel of minigene constructs was tested for autoregulation functionality, protein production and subcellular messenger RNA localization. Our data clearly indicate that constitutive spliceosome complex formation across intron 7 does not lead to high protein production but, on the contrary, to lower TDP-43 messenger RNA and protein levels. This is due to altered nucleocytoplasmic distribution of the RNA that is mostly retained in the nucleus and degraded. This study provides a novel in-depth characterization of how RNA binding proteins can autoregulate their own levels within cells, an essential regulatory process in maintaining cellular viability. PMID:24369426

Bembich, Sara; Herzog, Jeremias S; De Conti, Laura; Stuani, Cristiana; Avendaño-Vázquez, S Eréndira; Buratti, Emanuele; Baralle, Marco; Baralle, Francisco E

2014-03-01

 
 
 
 
121

Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation  

Science.gov (United States)

TDP-43 is a nuclear protein involved in many aspects of RNA metabolism. To ensure cellular viability, its expression levels within cells must be tightly regulated. We have previously demonstrated that TDP-43 autoregulation occurs through the activation of a normally silent intron in its 3?-UTR sequence that results in the use of alternative polyadenylation sites. In this work, we analyse which is the dominant event in autoregulation: the recognition of the splice sites of 3?-UTR intron 7 or the intrinsic quality of the alternative polyadenylation sites. A panel of minigene constructs was tested for autoregulation functionality, protein production and subcellular messenger RNA localization. Our data clearly indicate that constitutive spliceosome complex formation across intron 7 does not lead to high protein production but, on the contrary, to lower TDP-43 messenger RNA and protein levels. This is due to altered nucleocytoplasmic distribution of the RNA that is mostly retained in the nucleus and degraded. This study provides a novel in-depth characterization of how RNA binding proteins can autoregulate their own levels within cells, an essential regulatory process in maintaining cellular viability.

Bembich, Sara; Herzog, Jeremias S.; De Conti, Laura; Stuani, Cristiana; Avendano-Vazquez, S. Erendira; Buratti, Emanuele; Baralle, Marco; Baralle, Francisco E.

2014-01-01

122

Wild Type TDP-43 Induces Neuro-Inflammation and Alters APP Metabolism in Lentiviral Gene Transfer Models  

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The transactivation DNA-binding protein (TDP-43) pathology is associated with Fronto-Temporal Lobar Dementia (FTLD) with ubiquitinated inclusions and some cases of Alzheimer's disease (AD). Proteolytic fragments of ?-amyloid precursor protein (?APP) are detected in AD as well as the cerebrospinal fluid (CSF) from FTLD and Amyotrophic Lateral Sclerosis (ALS) patients, suggesting alteration in APP processing. Because of the overlap in TDP-43 pathology between FTLD and AD, we sought to determi...

Herman, Alexander M.; Khandelwal, Preeti J.; Rebeck, G. William; Moussa, Charbel E-h

2012-01-01

123

Cytoplasmic mislocalization of TDP-43 is toxic to neurons and enhanced by a mutation associated with familial ALS  

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Mutations in the gene encoding TDP-43 — the major protein component of neuronal aggregates characteristic of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with ubiquitin-positive inclusion bodies (FTLDu) — have been linked to familial forms of both disorders. Aggregates of TDP-43 in cortical and spinal motoneurons in ALS, or in neurons of the frontal and temporal cortices in FTLD, are closely linked to neuron loss and atrophy in these areas. However, the mechan...

Barmada, Sami J.; Skibinski, Gaia; Korb, Erica; Rao, Elizabeth J.; Wu, Jane Y.; Finkbeiner, Steven

2010-01-01

124

Age-dependent changes in TDP-43 levels in a mouse model of Alzheimer disease are linked to A? oligomers accumulation  

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Full Text Available Abstract Background Transactive response DNA-binding protein 43 (TDP-43 is the pathological protein found in frontotemporal lobar degeneration with ubiquitin positive inclusions and in amyotrophic lateral sclerosis. In diseased tissue, TDP-43 translocates from its physiological nuclear location into the cytoplasm, where it accumulates. Additionally, C-terminal fragments of TDP-43 accumulate in affected brain regions and are sufficient to cause TDP-43 mislocalization and cytoplasmic accumulation in vitro. TDP-43 also accumulates in 30% of Alzheimer disease (AD cases, a finding that has been highly reproducible. The role of TDP-43 in AD and its relation with A? and tau pathology, the two neuropathological hallmarks of AD, remains to be elucidated. Results Here we show that levels of TDP-43 and its ~35 kDa C-terminal fragment are significantly increased in the 3×Tg-AD mice, an animal model of AD that develops an age-dependent cognitive decline linked to the accumulation of A? and tau. We also report that the levels of TDP-43 and its C-terminal fragment correlate with the levels of soluble A? oligomers, which play a key role in AD pathogenesis. Notably, genetically reducing A?42 production restores the levels of TDP-43 and its ~35 kDa C-terminal fragment to control levels. Conclusions These data suggest a possible relation between A? oligomers and TDP-43.

Oddo Salvatore

2010-11-01

125

Neurotoxic 43-kDa TAR DNA-binding Protein (TDP-43) Triggers Mitochondrion-dependent Programmed Cell Death in Yeast*  

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Pathological neuronal inclusions of the 43-kDa TAR DNA-binding protein (TDP-43) are implicated in dementia and motor neuron disorders; however, the molecular mechanisms of the underlying cell loss remain poorly understood. Here we used a yeast model to elucidate cell death mechanisms upon expression of human TDP-43. TDP-43-expressing cells displayed markedly increased markers of oxidative stress, apoptosis, and necrosis. Cytotoxicity was dose- and age-dependent and was potentiated upon expres...

Braun, Ralf J.; Sommer, Cornelia; Carmona-gutierrez, Didac; Khoury, Chamel M.; Ring, Julia; Bu?ttner, Sabrina; Madeo, Frank

2011-01-01

126

Astrocyte pathology and the absence of non-cell autonomy in an induced pluripotent stem cell model of TDP-43 proteinopathy  

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Glial proliferation and activation are associated with disease progression in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar dementia. In this study, we describe a unique platform to address the question of cell autonomy in transactive response DNA-binding protein (TDP-43) proteinopathies. We generated functional astroglia from human induced pluripotent stem cells carrying an ALS-causing TDP-43 mutation and show that mutant astrocytes exhibit increased levels of TDP-43, subcellu...

Serio, Andrea; Bilican, Bilada; Barmada, Sami J.; Ando, Dale Michael; Zhao, Chen; Siller, Rick; Burr, Karen; Haghi, Ghazal; Story, David; Nishimura, Agnes Lumi; Carrasco, Monica A.; Phatnani, Hemali P.; Shum, Carole; Wilmut, Ian; Maniatis, Tom

2013-01-01

127

Cognitive Decline Typical of Frontotemporal Lobar Degeneration in Transgenic Mice Expressing the 25-kDa C-Terminal Fragment of TDP-43  

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Transactive response DNA-binding protein 43 (TDP-43) is the pathological signature protein in several neurodegenerative disorders, including the majority of frontotemporal lobar degeneration cases (FTLD-TDP), motor neuron disease, and amyotrophic lateral sclerosis. Pathological TDP-43 is mislocalized from its nuclear location to the cytoplasm, where it accumulates and is proteolytically cleaved to form C-terminal fragments. Although the 25-kDa C-terminal fragment of TDP-43 (TDP-25) accumulate...

Caccamo, Antonella; Majumder, Smita; Oddo, Salvatore

2012-01-01

128

TDP-43 Regulates Drosophila Neuromuscular Junctions Growth by Modulating Futsch/MAP1B Levels and Synaptic Microtubules Organization  

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TDP-43 is an evolutionarily conserved RNA binding protein recently associated with the pathogenesis of different neurological diseases. At the moment, neither its physiological role in vivo nor the mechanisms that may lead to neurodegeneration are well known. Previously, we have shown that TDP-43 mutant flies presented locomotive alterations and structural defects at the neuromuscular junctions. We have now investigated the functional mechanism leading to these phenotypes by screening several factors known to be important for synaptic growth or bouton formation. As a result we found that alterations in the organization of synaptic microtubules correlate with reduced protein levels in the microtubule associated protein futsch/MAP1B. Moreover, we observed that TDP-43 physically interacts with futsch mRNA and that its RNA binding capacity is required to prevent futsch down regulation and synaptic defects.

Romano, Maurizio; Appocher, Chiara; Klima, Raffaella; Buratti, Emanuele; Baralle, Francisco E.; Feiguin, Fabian

2011-01-01

129

The crystal structure of TDP-43 RRM1-DNA complex reveals the specific recognition for UG- and TG-rich nucleic acids.  

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TDP-43 is an important pathological protein that aggregates in the diseased neuronal cells and is linked to various neurodegenerative disorders. In normal cells, TDP-43 is primarily an RNA-binding protein; however, how the dimeric TDP-43 binds RNA via its two RNA recognition motifs, RRM1 and RRM2, is not clear. Here we report the crystal structure of human TDP-43 RRM1 in complex with a single-stranded DNA showing that RRM1 binds the nucleic acid extensively not only by the conserved ?-sheet residues but also by the loop residues. Mutational and biochemical assays further reveal that both RRMs in TDP-43 dimers participate in binding of UG-rich RNA or TG-rich DNA with RRM1 playing a dominant role and RRM2 playing a supporting role. Moreover, RRM1 of the amyotrophic lateral sclerosis-linked mutant D169G binds DNA as efficiently as the wild type; nevertheless, it is more resistant to thermal denaturation, suggesting that the resistance to degradation is likely linked to TDP-43 proteinopathies. Taken together all the data, we suggest a model showing that the two RRMs in each protomer of TDP-43 homodimer work together in RNA binding and thus the dimeric TDP-43 recognizes long clusters of UG-rich RNA to achieve high affinity and specificity. PMID:24464995

Kuo, Pan-Hsien; Chiang, Chien-Hao; Wang, Yi-Ting; Doudeva, Lyudmila G; Yuan, Hanna S

2014-04-01

130

Evolutionarily Conserved Heterogeneous Nuclear Ribonucleoprotein (hnRNP) A/B Proteins Functionally Interact with Human and Drosophila TAR DNA-binding Protein 43 (TDP-43).  

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Human TDP-43 represents the main component of neuronal inclusions found in patients with neurodegenerative diseases, especially frontotemporal lobar degeneration and amyotrophic lateral sclerosis. In vitro and in vivo studies have shown that the TAR DNA-binding protein 43 (TDP-43) Drosophila ortholog (TBPH) can biochemically and functionally overlap the properties of the human factor. The recent direct implication of the human heterogeneous nuclear ribonucleoproteins (hnRNPs) A2B1 and A1, known TDP-43 partners, in the pathogenesis of multisystem proteinopathy and amyotrophic lateral sclerosis supports the hypothesis that the physical and functional interplay between TDP-43 and hnRNP A/B orthologs might play a crucial role in the pathogenesis of neurodegenerative diseases. To test this hypothesis and further validate the fly system as a useful model to study this type of diseases, we have now characterized human TDP-43 and Drosophila TBPH similarity in terms of protein-protein interaction pathways. In this work we show that TDP-43 and TBPH share the ability to associate in vitro with Hrp38/Hrb98DE/CG9983, the fruit fly ortholog of the human hnRNP A1/A2 factors. Interestingly, the protein regions of TDP-43 and Hrp38 responsible for reciprocal interactions are conserved through evolution. Functionally, experiments in HeLa cells demonstrate that TDP-43 is necessary for the inhibitory activity of Hrp38 on splicing. Finally, Drosophila in vivo studies show that Hrp38 deficiency produces locomotive defects and life span shortening in TDP-43 with and without animals. These results suggest that hnRNP protein levels can play a modulatory role on TDP-43 functions. PMID:24492607

Romano, Maurizio; Buratti, Emanuele; Romano, Giulia; Klima, Raffaella; Del Bel Belluz, Lisa; Stuani, Cristiana; Baralle, Francisco; Feiguin, Fabian

2014-03-01

131

Astrocytes expressing mutant SOD1 and TDP43 trigger motoneuron death that is mediated via sodium channels and nitroxidative stress  

Science.gov (United States)

Amyotrophic lateral sclerosis (ALS) is a fatal paralytic disorder caused by dysfunction and degeneration of motor neurons. Multiple disease-causing mutations, including in the genes for SOD1 and TDP-43, have been identified in ALS. Astrocytes expressing mutant SOD1 are strongly implicated in the pathogenesis of ALS: we have shown that media conditioned by astrocytes carrying mutant SOD1G93A contains toxic factor(s) that kill motoneurons by activating voltage-sensitive sodium (Nav) channels. In contrast, a recent study suggests that astrocytes expressing mutated TDP43 contribute to ALS pathology, but do so via cell-autonomous processes and lack non-cell-autonomous toxicity. Here we investigate whether astrocytes that express diverse ALS-causing mutations release toxic factor(s) that induce motoneuron death, and if so, whether they do so via a common pathogenic pathway. We exposed primary cultures of wild-type spinal cord cells to conditioned medium derived from astrocytes (ACM) that express SOD1 (ACM-SOD1G93A and ACM-SOD1G86R) or TDP43 (ACM-TDP43A315T) mutants; we show that such exposure rapidly (within 30–60 min) increases dichlorofluorescein (DCF) fluorescence (indicative of nitroxidative stress) and leads to extensive motoneuron-specific death within a few days. Co-application of the diverse ACMs with anti-oxidants Trolox or esculetin (but not with resveratrol) strongly improves motoneuron survival. We also find that co-incubation of the cultures in the ACMs with Nav channel blockers (including mexiletine, spermidine, or riluzole) prevents both intracellular nitroxidative stress and motoneuron death. Together, our data document that two completely unrelated ALS models lead to the death of motoneuron via non-cell-autonomous processes, and show that astrocytes expressing mutations in SOD1 and TDP43 trigger such cell death through a common pathogenic pathway that involves nitroxidative stress, induced at least in part by Nav channel activity.

Rojas, Fabiola; Cortes, Nicole; Abarzua, Sebastian; Dyrda, Agnieszka; van Zundert, Brigitte

2013-01-01

132

Amyotrophic Lateral Sclerosis-associated Proteins TDP-43 and FUS/TLS Function in a Common Biochemical Complex to Co-regulate HDAC6 mRNA*  

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Amyotrophic lateral sclerosis (ALS) is an incurable neurodegenerative disease that preferentially targets motor neurons. It was recently found that dominant mutations in two related RNA-binding proteins, TDP-43 (43-kDa TAR DNA-binding domain protein) and FUS/TLS (fused in sarcoma/translated in liposarcoma) cause a subset of ALS. The convergent ALS phenotypes associated with TDP-43 and FUS/TLS mutations are suggestive of a functional relationship; however, whether or not TDP-43 and FUS/TLS ope...

Kim, Sang Hwa; Shanware, Naval P.; Bowler, Michael J.; Tibbetts, Randal S.

2010-01-01

133

Sensory and motor neuronopathy in a patient with the A382P TDP-43 mutation  

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Full Text Available Abstract Patients with TARDBP mutations have so far been classified as ALS, sometimes with frontal lobe dysfunction. A 66-year-old patient progressively developed a severe sensory disorder, followed by a motor disorder, which evolved over nine years. Symptoms started in the left hand and slowly involved the four limbs. Investigations were consistent with a mixed sensory and motor neuronopathy. A heterozygous change from an alanine to a proline at amino acid 382 was identified in exon 6 of the TARDPB gene (p.A382P. This case expands the phenotypic spectrum associated with mutations in the TARDBP gene and shows that sensory neurons can be severely damaged early in the course of the disease, following a propagating process, with an orderly progression from a focal starting point. A combination of severe sensory and motor neuronopathy is rarely encountered in clinical practice. The possibility of an A382P TDP-43 mutation should be considered in patients with such an association.

Rouleau Guy A

2011-02-01

134

?-N-methylamino-L-alanine induces changes in both GSK3 and TDP-43 in human neuroblastoma.  

Science.gov (United States)

?-N-methylamino-L-alanine (L-BMAA) is a neurotoxic amino acid produced by most cyanobacteria, which are extensively distributed in different environments all over the world. L-BMAA has been linked to a variety of neurodegenerative diseases. This work aims to analyze the toxicological action of L-BMAA related to alterations observed in different neurodegenerative illness as Alzheimer disease and amyotrophic lateral sclerosis. Our results demonstrate that neuroblastoma cells treated with L-BMAA show an increase in glycogen synthase kinase 3 ? (GSk3?) and induce accumulation of TAR DNA-binding protein 43 (TDP-43) truncated forms (C-terminal fragments), phosphorylated  and high molecular weight forms of TDP-43, that appears frequently in some neurodegenerative diseases. PMID:23665941

Muñoz-Saez, Emma; de Munck, Estefanía; Arahuetes, Rosa M; Solas, M Teresa; Martínez, Ana M; Miguel, Begoña G

2013-01-01

135

The C-Terminal TDP-43 Fragments Have a High Aggregation Propensity and Harm Neurons by a Dominant-Negative Mechanism  

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TAR DNA binding protein 43 KD (TDP-43) is an essential gene that regulates gene transcription, mRNA splicing and stability. In amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two fatal neurodegenerative diseases, TDP-43 is fragmented, generating multiple fragments that include the C-terminal fragment of ?25 KD. The role of these fragments in the pathogenesis of ALS and FTD is not clear. Here we investigated the aggregation propensity in various polypeptide regions of ...

Yang, Chunxing; Tan, Weijia; Whittle, Catheryne; Qiu, Linghua; Cao, Lucheng; Akbarian, Schahram; Xu, Zuoshang

2010-01-01

136

Astrocytes expressing mutant SOD1 and TDP43 trigger motoneuron death that is mediated via sodium channels and nitroxidative stress  

Directory of Open Access Journals (Sweden)

Full Text Available Amyotrophic lateral sclerosis (ALS is a fatal paralytic disorder caused by dysfunction and degeneration of motor neurons. Multiple disease-causing mutations, including in the genes for SOD1 and TDP-43, have been identified in ALS. Astrocytes expressing mutant SOD1 are strongly implicated in the pathogenesis of ALS: we have shown that media conditioned by astrocytes carrying mutant SOD1G93A contains toxic factor(s that kill motoneurons by activating voltage-sensitive sodium (Nav channels. In contrast, a recent study suggests that astrocytes expressing mutated TDP43 contribute to ALS pathology, but do so via cell-autonomous processes and lack non-cell-autonomous toxicity. Here we investigate whether astrocytes that express diverse ALS-causing mutations release toxic factor(s that induce motoneuron death, and if so, whether they do so via a common pathogenic pathway. We exposed primary cultures of wild-type spinal cord cells to conditioned medium derived from astrocytes (ACM that express SOD1 (ACM-SOD1G93A and ACM-SOD1G86R or TDP43 (ACM-TDP43A315T mutants; we show that such exposure rapidly (within 30-60 minutes increases dichlorofluorescein (DCF fluorescence (indicative of nitroxidative stress and leads to extensive motoneuron-specific death within a few days. Co-application of the diverse ACMs with anti-oxidants Trolox or esculetin (but not with resveratrol strongly improves motoneuron survival. We also find that co-incubation of the cultures in the ACMs with Nav channel blockers (including mexiletine, spermidine or riluzole prevents both intracellular nitroxidative stress and motoneuron death. Together, our data document that two completely unrelated ALS models lead to the death of motoneuron via non-cell-autonomous processes, and show that astrocytes expressing mutations in SOD1 and TDP43 trigger such cell death through a common pathogenic pathway that involves nitroxidative stress, induced at least in part by Nav channel activity.

BrigitteA.J.Van Zundert

2014-02-01

137

Self-assembling properties of peptides derived from TDP-43 C-terminal fragment.  

Science.gov (United States)

Two highly fibrillogenic peptide sequences (MNFGAFSINP and EDLIIKGISV) were previously reported in the C-terminal fragment (CTF) of TDP-43 (220-414), a protein recently implicated in neuro-degenerative diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD-U). It was observed that the sequences MNFGAFS and EDLIIKG harbor their respective fibrillogenic domains. Here, the self-assembling properties of peptides obtained by systematic deletion of residues from these two sequences were investigated with the help of light scattering, thioflavin T fluorescence, transmission electron microscopy, and circular dichroism spectroscopy. It was found that the pentapeptide NFGAF and the tetrapeptide DLII are the shortest fibrillogenic sequences from MNFGAFS and EDLIIKG, respectively. Structure function studies revealed that self-assembly of the peptides is largely governed by hydrophobic interactions. Both NFGAF and DLII formed hydrogels based on a complex fibrillar network, at relatively low concentrations, and of remarkable strength and stability. Of particular interest was DLII, a rare aliphatic tetrapeptide that formed a hydrogel at a concentration of 1 mg/mL in less than an hour. Interestingly, various other tetrapeptides based on DLII (YLII, KLII, NLII, and LIID) also formed hydrogels of comparable physical properties, suggesting that an amphipathic peptide design based on the hydrophobic LII motif and a single residue polar terminus is highly favorable for hydrogelation. Peptides discovered in this study, especially DLII and its variants, are some of the shortest ever reported to show such structural and functional features, suggesting that they can be useful templates for the design of peptide-based soft materials. PMID:24559403

Saini, Akash; Chauhan, Virander S

2014-04-01

138

Increased metal content in the TDP-43(A315T) transgenic mouse model of frontotemporal lobar degeneration and amyotrophic lateral sclerosis.  

Science.gov (United States)

Disrupted metal homeostasis is a consistent feature of neurodegenerative disease in humans and is recapitulated in mouse models of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS) and neuronal ceriod lipofuscinosis. While the definitive pathogenesis of neurodegenerative disease in humans remains to be fully elucidated, disease-like symptoms in the mouse models are all driven by the presence or over-expression of a putative pathogenic protein, indicating an in vivo relationship between expression of these proteins, disrupted metal homeostasis and the symptoms of neuronal failure. Recently it was established that mutant TAR DNA binding protein-43 (TDP-43) is associated with the development of frontotemporal lobar degeneration and ALS. Subsequent development of transgenic mice that express human TDP-43 carrying the disease-causing A315T mutation has provided new opportunity to study the underlying mechanisms of TDP-43-related neurodegenerative disease. We assessed the cognitive and locomotive phenotype of TDP-43 (A315T) mice and their wild-type littermates and also assessed bulk metal content of brain and spinal cord tissues. Metal levels in the brain were not affected by the expression of mutant TDP-43, but zinc, copper, and manganese levels were all increased in the spinal cords of TDP-43 (A315T) mice when compared to wild-type littermates. Performance of the TDP-43 (A315T) mice in the Y-maze test for cognitive function was not significantly different to wild-type mice. By contrast, performance of the TDP-43 (A315T) in the rotarod test for locomotive function was consistently worse than wild-type mice. These preliminary in vivo data are the first to show that expression of a disease-causing form of TDP-43 is sufficient to disrupt metal ion homeostasis in the central nervous system. Disrupted metal ion homeostasis in the spinal cord but not the brain may explain why the TDP-43 (A315T) mice show symptoms of locomotive decline and not cognitive decline. PMID:24575040

Dang, Theresa N T; Lim, Nastasia K H; Grubman, Alexandra; Li, Qiao-Xin; Volitakis, Irene; White, Anthony R; Crouch, Peter J

2014-01-01

139

Increased metal content in the TDP-43A315T transgenic mouse model of frontotemporal lobar degeneration and amyotrophic lateral sclerosis  

Directory of Open Access Journals (Sweden)

Full Text Available Disrupted metal homeostasis is a consistent feature of neurodegenerative disease in humans and is recapitulated in mouse models of Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis (ALS and neuronal ceriod lipofuscinosis. While the definitive pathogenesis of neurodegenerative disease in humans remains to be fully elucidated, disease-like symptoms in the mouse models are all driven by the presence or over-expression of a putative pathogenic protein, indicating an in vivo relationship between expression of these proteins, disrupted metal homeostasis and the symptoms of neuronal failure. Recently it was established that mutant TAR DNA binding protein-43 (TDP-43 is associated with the development of frontotemporal lobar degeneration (FTLD and ALS. Subsequent development of transgenic mice that express human TDP-43 carrying the disease-causing A315T mutation has provided new opportunity to study the underlying mechanisms of TDP-43-related neurodegenerative disease. We assessed the cognitive and locomotive phenotype of TDP-43A315T mice and their wild-type littermates and also assessed bulk metal content of brain and spinal cord tissues. Metal levels in the brain were not affected by the expression of mutant TDP-43, but Zn, Cu and Mn levels were all increased in the spinal cords of TDP-43A315T mice when compared to wild-type littermates. Performance of the TDP-43A315T mice in the Y-maze test for cognitive function was not significantly different to wild-type mice. By contrast, performance of the TDP-43A315T in the rotarod test for locomotive function was consistently worse than wild-type mice. These preliminary in vivo data are the first to show that expression of a disease-causing form of TDP-43 is sufficient to disrupt metal ion homeostasis in the central nervous system. Disrupted metal ion homeostasis in the spinal cord but not the brain may explain why the TDP-43A315T mice show symptoms of locomotive decline and not cognitive decline.

PeterCrouch

2014-02-01

140

Increased metal content in the TDP-43A315T transgenic mouse model of frontotemporal lobar degeneration and amyotrophic lateral sclerosis  

Science.gov (United States)

Disrupted metal homeostasis is a consistent feature of neurodegenerative disease in humans and is recapitulated in mouse models of Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis (ALS) and neuronal ceriod lipofuscinosis. While the definitive pathogenesis of neurodegenerative disease in humans remains to be fully elucidated, disease-like symptoms in the mouse models are all driven by the presence or over-expression of a putative pathogenic protein, indicating an in vivo relationship between expression of these proteins, disrupted metal homeostasis and the symptoms of neuronal failure. Recently it was established that mutant TAR DNA binding protein-43 (TDP-43) is associated with the development of frontotemporal lobar degeneration and ALS. Subsequent development of transgenic mice that express human TDP-43 carrying the disease-causing A315T mutation has provided new opportunity to study the underlying mechanisms of TDP-43-related neurodegenerative disease. We assessed the cognitive and locomotive phenotype of TDP-43 A315T mice and their wild-type littermates and also assessed bulk metal content of brain and spinal cord tissues. Metal levels in the brain were not affected by the expression of mutant TDP-43, but zinc, copper, and manganese levels were all increased in the spinal cords of TDP-43 A315T mice when compared to wild-type littermates. Performance of the TDP-43 A315T mice in the Y-maze test for cognitive function was not significantly different to wild-type mice. By contrast, performance of the TDP-43 A315T in the rotarod test for locomotive function was consistently worse than wild-type mice. These preliminary in vivo data are the first to show that expression of a disease-causing form of TDP-43 is sufficient to disrupt metal ion homeostasis in the central nervous system. Disrupted metal ion homeostasis in the spinal cord but not the brain may explain why the TDP-43 A315T mice show symptoms of locomotive decline and not cognitive decline.

Dang, Theresa N. T.; Lim, Nastasia K. H.; Grubman, Alexandra; Li, Qiao-Xin; Volitakis, Irene; White, Anthony R.; Crouch, Peter J.

2014-01-01

 
 
 
 
141

ApoE and TDP-43 neuropathology in two siblings with familial FTLD-motor neuron disease  

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Frontotemporal lobar degeneration with motor neuron disease (FTLD-MND) is characterized by neuronal cytoplasmic inclusions containing TDP-43. Apolipoprotein E4 (apoE4), derived from the apoE ?4 allele, enhances brain atrophy in FTLD through unknown mechanisms. Here, we studied two siblings with C9ORF72-linked familial FTLD-MND, an apoE ?4 homozygote and an apoE ?3 homozygote. The apoE ?4 homozygote had more cognitive-behavioral symptoms, fronto-insulo-temporal atrophy, and apoE fragments ...

2013-01-01

142

The RNA-binding Protein TDP-43 Selectively Disrupts MicroRNA-1/206 Incorporation into the RNA-induced Silencing Complex*?  

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MicroRNA (miRNA) maturation is regulated by interaction of particular miRNA precursors with specific RNA-binding proteins. Following their biogenesis, mature miRNAs are incorporated into the RNA-induced silencing complex (RISC) where they interact with mRNAs to negatively regulate protein production. However, little is known about how mature miRNAs are regulated at the level of their activity. To address this, we screened for proteins differentially bound to the mature form of the miR-1 or miR-133 miRNA families. These muscle-enriched, co-transcribed miRNA pairs cooperate to suppress smooth muscle gene expression in the heart. However, they also have opposing roles, with the miR-1 family, composed of miR-1 and miR-206, promoting myogenic differentiation, whereas miR-133 maintains the progenitor state. Here, we describe a physical interaction between TDP-43, an RNA-binding protein that forms aggregates in the neuromuscular disease, amyotrophic lateral sclerosis, and the miR-1, but not miR-133, family. Deficiency of the TDP-43 Drosophila ortholog enhanced dmiR-1 activity in vivo. In mammalian cells, TDP-43 limited the activity of both miR-1 and miR-206, but not the miR-133 family, by disrupting their RISC association. Consistent with TDP-43 dampening miR-1/206 activity, protein levels of the miR-1/206 targets, IGF-1 and HDAC4, were elevated in TDP-43 transgenic mouse muscle. This occurred without corresponding Igf-1 or Hdac4 mRNA increases and despite higher miR-1 and miR-206 expression. Our findings reveal that TDP-43 negatively regulates the activity of the miR-1 family of miRNAs by limiting their bioavailability for RISC loading and suggest a processing-independent mechanism for differential regulation of miRNA activity.

King, Isabelle N.; Yartseva, Valeria; Salas, Donaldo; Kumar, Abhishek; Heidersbach, Amy; Ando, D. Michael; Stallings, Nancy R.; Elliott, Jeffrey L.; Srivastava, Deepak; Ivey, Kathryn N.

2014-01-01

143

ER-mitochondria associations are regulated by the VAPB-PTPIP51 interaction and are disrupted by ALS/FTD-associated TDP-43  

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Mitochondria and the endoplasmic reticulum (ER) form tight structural associations and these facilitate a number of cellular functions. However, the mechanisms by which regions of the ER become tethered to mitochondria are not properly known. Understanding these mechanisms is not just important for comprehending fundamental physiological processes but also for understanding pathogenic processes in some disease states. In particular, disruption to ER–mitochondria associations is linked to some neurodegenerative diseases. Here we show that the ER-resident protein VAPB interacts with the mitochondrial protein tyrosine phosphatase-interacting protein-51 (PTPIP51) to regulate ER–mitochondria associations. Moreover, we demonstrate that TDP-43, a protein pathologically linked to amyotrophic lateral sclerosis and fronto-temporal dementia perturbs ER–mitochondria interactions and that this is associated with disruption to the VAPB–PTPIP51 interaction and cellular Ca2+ homeostasis. Finally, we show that overexpression of TDP-43 leads to activation of glycogen synthase kinase-3? (GSK-3?) and that GSK-3? regulates the VAPB–PTPIP51 interaction. Our results describe a new pathogenic mechanism for TDP-43.

Stoica, Radu; De Vos, Kurt J.; Paillusson, Sebastien; Mueller, Sarah; Sancho, Rosa M.; Lau, Kwok-Fai; Vizcay-Barrena, Gema; Lin, Wen-Lang; Xu, Ya-Fei; Lewis, Jada; Dickson, Dennis W.; Petrucelli, Leonard; Mitchell, Jacqueline C.; Shaw, Christopher E.; Miller, Christopher C. J.

2014-01-01

144

The RNA-binding motif 45 (RBM45) protein accumulates in inclusion bodies in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with TDP-43 inclusions (FTLD-TDP) patients  

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RNA-binding protein pathology now represents one of the best characterized pathologic features of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration patients with TDP-43 or FUS pathology (FTLD-TDP and FTLD-FUS). Using liquid chromatography tandem mass spectrometry, we identified altered levels of the RNA-binding motif 45 (RBM45) protein in the cerebrospinal fluid (CSF) of ALS patients. This protein contains sequence similarities to TAR DNA-binding protein 43 (TDP-43) an...

2012-01-01

145

An early onset progressive motor neuron disorder in Scyl1-deficient mice is associated with mislocalization of TDP-43.  

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The molecular and cellular bases of motor neuron diseases (MNDs) are still poorly understood. The diseases are mostly sporadic, with ~10% of cases being familial. In most cases of familial motor neuronopathy, the disease is caused by either gain-of-adverse-effect mutations or partial loss-of-function mutations in ubiquitously expressed genes that serve essential cellular functions. Here we show that deletion of Scyl1, an evolutionarily conserved and ubiquitously expressed gene encoding the COPI-associated protein pseudokinase SCYL1, causes an early onset progressive MND with characteristic features of amyotrophic lateral sclerosis (ALS). Skeletal muscles of Scyl1(-/-) mice displayed neurogenic atrophy, fiber type switching, and disuse atrophy. Peripheral nerves showed axonal degeneration. Loss of lower motor neurons (LMNs) and large-caliber axons was conspicuous in Scyl1(-/-) animals. Signs of neuroinflammation were seen throughout the CNS, most notably in the ventral horn of the spinal cord. Neural-specific, but not skeletal muscle-specific, deletion of Scyl1 was sufficient to cause motor dysfunction, indicating that SCYL1 acts in a neural cell-autonomous manner to prevent LMN degeneration and motor functions. Remarkably, deletion of Scyl1 resulted in the mislocalization and accumulation of TDP-43 (TAR DNA-binding protein of 43 kDa) and ubiquilin 2 into cytoplasmic inclusions within LMNs, features characteristic of most familial and sporadic forms of ALS. Together, our results identify SCYL1 as a key regulator of motor neuron survival, and Scyl1(-/-) mice share pathological features with many human neurodegenerative conditions. PMID:23175812

Pelletier, Stephane; Gingras, Sebastien; Howell, Sherie; Vogel, Peter; Ihle, James N

2012-11-21

146

The voltage-gated calcium channel blocker lomerizine is neuroprotective in motor neurons expressing mutant SOD1, but not TDP-43.  

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Excitotoxicity and disruption of Ca(2+) homeostasis have been implicated in amyotrophic lateral sclerosis (ALS) and limiting Ca(2+) entry is protective in models of ALS caused by mutation of SOD1. Lomerizine, an antagonist of L- and T-type voltage-gated calcium channels and transient receptor potential channel 5 transient receptor potential channels, is well tolerated clinically, making it a potential therapeutic candidate. Lomerizine reduced glutamate excitotoxicity in cultured motor neurons by reducing the accumulation of cytoplasmic Ca(2+) and protected motor neurons against multiple measures of mutant SOD1 toxicity: Ca(2+) overload, impaired mitochondrial trafficking, mitochondrial fragmentation, formation of mutant SOD1 inclusions, and loss of viability. To assess the utility of lomerizine in other forms of ALS, calcium homeostasis was evaluated in culture models of disease because of mutations in the RNA-binding proteins transactive response DNA-binding protein 43 (TDP-43) and Fused in Sarcoma (FUS). Calcium did not play the same role in the toxicity of these mutant proteins as with mutant SOD1 and lomerizine failed to prevent cytoplasmic accumulation of mutant TDP-43, a hallmark of its pathology. These experiments point to differences in the pathogenic pathways between types of ALS and show the utility of primary culture models in comparing those mechanisms and effectiveness of therapeutic strategies. Calcium sensitivity is a factor in motor neuron vulnerability in ALS. The voltage-gated calcium channel blocker lomerizine normalized [Ca(2+) ] and reduced toxicity of mutant Cu/Zn-superoxide dismutase (SOD1) causing familial ALS1. Calcium homeostasis was not disrupted in motor neurons expressing ALS-associated mutants of TAR DNA-binding protein 43 (TDP-43) or FUS, nor was lomerizine protective, affirming differences in pathogenic mechanism and therapeutic efficacy in the forms of ALS. PMID:24716897

Tran, Luan T; Gentil, Benoit J; Sullivan, Kathleen E; Durham, Heather D

2014-08-01

147

Common variation in the miR-659 binding-site of GRN is a major risk factor for TDP43-positive frontotemporal dementia  

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Loss-of-function mutations in progranulin (GRN) cause ubiquitin- and TAR DNA-binding protein 43 (TDP-43)-positive frontotemporal dementia (FTLD-U), a progressive neurodegenerative disease affecting ?10% of early-onset dementia patients. Here we expand the role of GRN in FTLD-U and demonstrate that a common genetic variant (rs5848), located in the 3?-untranslated region (UTR) of GRN in a binding-site for miR-659, is a major susceptibility factor for FTLD-U. In a series of pathologically co...

Rademakers, Rosa; Eriksen, Jason L.; Baker, Matt; Robinson, Todd; Ahmed, Zeshan; Lincoln, Sarah J.; Finch, Nicole; Rutherford, Nicola J.; Crook, Richard J.; Josephs, Keith A.; Boeve, Bradley F.; Knopman, David S.; Petersen, Ronald C.; Parisi, Joseph E.; Caselli, Richard J.

2008-01-01

148

A Novel GRN Mutation (GRN c.708+6_+9delTGAG) in Frontotemporal Lobar Degeneration With TDP-43-Positive Inclusions: Clinicopathologic Report of 6 Cases.  

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Understanding of frontotemporal lobar degeneration, the underlying pathology most often linked to the clinical diagnosis of frontotemporal dementia, is rapidly increasing. Mutations in 7 known genes (MAPT, GRN, C9orf72, VCP, CHMP2B, and, rarely, TARDBP and FUS) are associated with frontotemporal dementia, and the pathologic classification of frontotemporal lobar degeneration has recently been modified to reflect these discoveries. Mutations in one of these genes (GRN), which encodes progranulin, have been implicated in up to a quarter of cases of frontotemporal lobar degeneration with TDP-43 (TAR DNA-binding protein 43)-positive inclusions; currently, there are more than 60 known pathogenic mutations of the gene. We present the clinical, pathologic, and genetic findings on 6 cases from 4 families, 5 of which were shown to have a novel GRN c.708+6_+9delTGAG mutation. PMID:24709683

Bit-Ivan, Esther N; Suh, Eunran; Shim, Hyung-Sub; Weintraub, Sandra; Hyman, Bradley T; Arnold, Steven E; McCarty-Wood, Elisabeth; Van Deerlin, Viviana M; Schneider, Julie A; Trojanowski, John Q; Frosch, Matthew P; Baker, Matt C; Rademakers, Rosa; Mesulam, Marsel; Bigio, Eileen H

2014-05-01

149

FTLD-TDP with motor neuron disease, visuospatial impairment and a progressive supranuclear palsy-like syndrome: broadening the clinical phenotype of TDP-43 proteinopathies. A report of three cases  

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Abstract Background Frontotemporal lobar degeneration with ubiquitin and TDP-43 positive neuronal inclusions represents a novel entity (FTLD-TDP) that may be associated with motor neuron disease (FTLD-MND); involvement of extrapyramidal and other systems has also been reported. Case presentation We present three cases with similar clinical symptoms, including Parkinsonism, supranuclear gaze palsy, visuospatial impairment and a behavioral variant of frontotempora...

2011-01-01

150

Altered microRNA expression in frontotemporal lobar degeneration with TDP-43 pathology caused by progranulin mutations  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Frontotemporal lobar degeneration (FTLD is a progressive neurodegenerative disorder that can be triggered through genetic or sporadic mechanisms. MicroRNAs (miRNAs have become a major therapeutic focus as their pervasive expression and powerful regulatory roles in disease pathogenesis become increasingly apparent. Here we examine the role of miRNAs in FTLD patients with TAR DNA-binding protein 43 pathology (FTLD-TDP caused by genetic mutations in the progranulin (PGRN gene. Results Using miRNA array profiling, we identified the 20 miRNAs that showed greatest evidence (unadjusted P PGRN mutations when compared to 32 FTLD-TDP patients with no apparent genetic abnormalities. Quantitative real-time PCR (qRT-PCR analyses provided technical validation of the differential expression for 9 of the 20 miRNAs in frontal cortex. Additional qRT-PCR analyses showed that 5 out of 9 miRNAs (miR-922, miR-516a-3p, miR-571, miR-548b-5p, and miR-548c-5p were also significantly dysregulated (unadjusted P PGRN mutation carriers, consistent with a systemic reduction in PGRN levels. We developed a list of gene targets for the 5 candidate miRNAs and found 18 genes dysregulated in a reported FTLD mRNA study to exhibit anti-correlated miRNA-mRNA patterns in affected cortex and cerebellar tissue. Among the targets is brain-specific angiogenesis inhibitor 3, which was recently identified as an important player in synapse biology. Conclusions Our study suggests that miRNAs may contribute to the pathogenesis of FTLD-TDP caused by PGRN mutations and provides new insight into potential future therapeutic options.

Kocerha Jannet

2011-10-01

151

Common variation in the miR-659 binding-site of GRN is a major risk factor for TDP43-positive frontotemporal dementia  

Science.gov (United States)

Loss-of-function mutations in progranulin (GRN) cause ubiquitin- and TAR DNA-binding protein 43 (TDP-43)-positive frontotemporal dementia (FTLD-U), a progressive neurodegenerative disease affecting ?10% of early-onset dementia patients. Here we expand the role of GRN in FTLD-U and demonstrate that a common genetic variant (rs5848), located in the 3?-untranslated region (UTR) of GRN in a binding-site for miR-659, is a major susceptibility factor for FTLD-U. In a series of pathologically confirmed FTLD-U patients without GRN mutations, we show that carriers homozygous for the T-allele of rs5848 have a 3.2-fold increased risk to develop FTLD-U compared with homozygous C-allele carriers (95% CI: 1.50–6.73). We further demonstrate that miR-659 can regulate GRN expression in vitro, with miR-659 binding more efficiently to the high risk T-allele of rs5848 resulting in augmented translational inhibition of GRN. A significant reduction in GRN protein was observed in homozygous T-allele carriers in vivo, through biochemical and immunohistochemical methods, mimicking the effect of heterozygous loss-of-function GRN mutations. In support of these findings, the neuropathology of homozygous rs5848 T-allele carriers frequently resembled the pathological FTLD-U subtype of GRN mutation carriers. We suggest that the expression of GRN is regulated by miRNAs and that common genetic variability in a miRNA binding-site can significantly increase the risk for FTLD-U. Translational regulation by miRNAs may represent a common mechanism underlying complex neurodegenerative disorders.

Rademakers, Rosa; Eriksen, Jason L.; Baker, Matt; Robinson, Todd; Ahmed, Zeshan; Lincoln, Sarah J.; Finch, Nicole; Rutherford, Nicola J.; Crook, Richard J.; Josephs, Keith A.; Boeve, Bradley F.; Knopman, David S.; Petersen, Ronald C.; Parisi, Joseph E.; Caselli, Richard J.; Wszolek, Zbigniew K.; Uitti, Ryan J.; Feldman, Howard; Hutton, Michael L.; Mackenzie, Ian R.; Graff-Radford, Neill R.; Dickson, Dennis W.

2008-01-01

152

Abnormal distribution of heterogeneous nuclear ribonucleoproteins in sporadic inclusion body myositis.  

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Previous histopathologic studies of sporadic inclusion body myositis (sIBM) identified sarcoplasmic aggregation and myonuclear depletion of the predominantly nuclear heterogeneous nuclear ribonucleoprotein (hnRNP) TDP-43 in sIBM myofibers. Here, we examined sIBM muscle for abnormalities in two other hnRNPs hnRNPA1 and hnRNPA2B1, mutations in which cause multisystem proteinopathy associated with rimmed-vacuolar myopathies. Muscle biopsy specimens from 13 patients with sIBM and 13 patients without sIBM (dermatomyositis N=3, polymyositis N=3, muscular dystrophy N=3, motor neuron disease N=2, non-neuromuscular disease N=2) underwent immunohistochemistry for hnRNPA1, hnRNPA2B1, and TDP-43. Muscle transcriptional microarray data from 27 patients with sIBM and 12 patients without neuromuscular disease was analyzed. Depletion of hnRNPA1 and hnRNPA2B1 was present in 15% and 7% of sIBM myonuclei, respectively, compared with 1% and 0% of myonuclei in non-sIBM muscle. Sarcoplasmic aggregates of hnRNPA1 and hnRNPA2B1 distinct from TDP-43 aggregates were also found in sIBM. hnRNPA1 and hnRNPA2B1, as well as other hnRNPs, gene expression was unaltered in sIBM compared to normal muscle. Along with TDP-43, other hnRNPs, including hnRNPA1 and hnRNPA2B1, are depleted from sIBM myonuclei at the protein but not transcript level. The depletion of multiple hnRNPs from sIBM myonuclei together with their sarcoplasmic aggregation suggests that one aspect of sIBM pathophysiology may involve abnormal RNA metabolism that includes hyperassembly of ribonucleoprotein granules mediated by prion-like domains in hnRNPs, evolving into pathological aggregates. PMID:24857366

Pinkus, Jack L; Amato, Anthony A; Taylor, J Paul; Greenberg, Steven A

2014-07-01

153

Phosphorylation regulates proteasomal-mediated degradation and solubility of TAR DNA binding protein-43 C-terminal fragments  

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Full Text Available Abstract Background Inclusions of TAR DNA binding protein-43 (TDP-43 are the defining histopathological feature of several neurodegenerative diseases collectively referred to as TDP-43 proteinopathies. These diseases are characterized by the presence of cellular aggregates composed of abnormally phosphorylated, N-terminally truncated and ubiquitinated TDP-43 in the spinal cord and/or brain. Recent studies indicate that C-terminal fragments of TDP-43 are aggregation-prone and induce cytotoxicity. However, little is known regarding the pathways responsible for the degradation of these fragments and how their phosphorylation contributes to the pathogenesis of disease. Results Herein, we established a human neuroblastoma cell line (M17D3 that conditionally expresses an enhanced green fluorescent protein (GFP-tagged caspase-cleaved C-terminal TDP-43 fragment (GFP-TDP220-414. We report that expression of this fragment within cells leads to a time-dependent formation of inclusions that are immunoreactive for both ubiquitin and phosphorylated TDP-43, thus recapitulating pathological hallmarks of TDP-43 proteinopathies. Phosphorylation of GFP-TDP220-414 renders it resistant to degradation and enhances its accumulation into insoluble aggregates. Nonetheless, GFP-TDP220-414 inclusions are reversible and can be cleared through the ubiquitin proteasome system. Moreover, both Hsp70 and Hsp90 bind to GFP-TDP220-414 and regulate its degradation. Conclusions Our data indicates that inclusions formed from TDP-43 C-terminal fragments are reversible. Given that TDP-43 inclusions have been shown to confer toxicity, our findings have important therapeutic implications and suggest that modulating the phosphorylation state of TDP-43 C-terminal fragments may be a promising therapeutic strategy to clear TDP-43 inclusions.

Zhang Yong-Jie

2010-08-01

154

A Seeding Reaction Recapitulates Intracellular Formation of Sarkosyl-insoluble Transactivation Response Element (TAR) DNA-binding Protein-43 Inclusions*?  

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The transactivation response element (TAR) DNA-binding protein-43 (TDP-43) is a nuclear protein that normally regulates transcription and splicing. Abnormal accumulation of insoluble inclusions containing TDP-43 has been recently reported in the affected tissues of amyotrophic lateral sclerosis (ALS) patients. Here, we show that intracellular aggregation of TDP-43 can be triggered by transduction of fibrillar aggregates prepared from in vitro functional TDP-43. Sarkosyl is found to be incapab...

Furukawa, Yoshiaki; Kaneko, Kumi; Watanabe, Shoji; Yamanaka, Koji; Nukina, Nobuyuki

2011-01-01

155

Reduction in brain immunoreactive corticotropin-releasing factor (CRF) in spontaneously hypertensive rats  

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The brain CRF concentration of spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY) was examined by rat CRF radioimmunoassay. Anti-CRF serum was developed by immunizing rabbits with synthetic rat CRF. Synthetic rat CRF was also used as tracer and standard. The displacement of /sup 125/I-rat CRF by serially diluted extracts of male Wistar rats hypothalamus, thalamus, midbrain, pons, medulla oblongata, cerebral cortex, cerebellum and neurointermediate lobe was parallel to the displacement of synthetic rat CRF. In both WKY and SHR the highest levels of CRF immunoreactivity were shown by the hypothalamus and neurointermediate lobe, and considerable CRF immunoreactivity was also detected in other brain regions. The CRF immunoreactivity in the hypothalamus, neurointermediate lobe, midbrain, medulla oblongata and cerebral cortex was significantly reduced in SHR and it may suggest that CRF abnormality may be implicated in the reported abnormalities in the pituitary-adrenal axis, autonomic response and behavior of SHR.

Hashimoto, K.; Hattori, T.; Murakami, K.; Suemaru, S.; Kawada, Y.; Kageyama, J.; Ota, Z.

1985-02-18

156

Endogenous TDP-43, but not FUS, contributes to stress granule assembly via G3BP  

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Abstract Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the selective loss of upper and lower motor neurons, a cell type that is intrinsically more vulnerable than other cell types to exogenous stress. The interplay between genetic susceptibility and environmental exposures to toxins has long been thought to be relevant to ALS. One cellular mechanism to overcome stress is the formation of small dense cytoplasmic domains called stress granul...

Aulas Anaïs; Stabile Stéphanie; Vande Velde Christine

2012-01-01

157

Immunoreactivity of specific epitopes of PrPSc is enhanced by pretreatment in a hydrated autoclave.  

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An abnormal protein (PrPSc) accumulates in animals affected with scrapie. Immunoblotting procedures have been used widely to detect PrPSc. Blotted membranes were subjected to pretreatment in a hydrated autoclave, and the subsequent immunoreactivity of PrPSc was examined. The immunoreactivity of PrPSc to antisera against the synthetic peptides of the mouse PrP amino acid sequences 199 to 208 and 213 to 226 was enhanced by the pretreatment. However, the reactivity to antisera of peptide sequenc...

Yokoyama, T.; Momotani, E.; Kimura, K.; Yuasa, N.

1996-01-01

158

Meiotic abnormalities  

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Chapter 19, describes meiotic abnormalities. These include nondisjunction of autosomes and sex chromosomes, genetic and environmental causes of nondisjunction, misdivision of the centromere, chromosomally abnormal human sperm, male infertility, parental age, and origin of diploid gametes. 57 refs., 2 figs., 1 tab.

NONE

1993-12-31

159

Walking abnormalities  

Science.gov (United States)

... include: Arthritis of the leg or foot joints Conversion disorder (a psychological disorder) Foot problems (such as a ... injuries. For an abnormal gait that occurs with conversion disorder, counseling and support from family members are strongly ...

160

Polyamine-like immunoreactivity in rat neurons.  

Science.gov (United States)

The localization of polyamine (PA) pools in motor, sensory, and autonomic neurons and in the nerve cells of the hypothalamo-hypophysial system of rats was examined by immunocytochemical method using the monoclonal antibody ASPM-29 specific to spermine (Spm) and spermidine (Spd) fixed in situ. Strong PA immunoreactivity was found in the cytoplasm and dendrites of the large perikaryon of motor neurons in the anterior spinal column, in the Purkinje cells of the cerebellum, in the pyramidal cells of the cerebrum, in the nerve cells of the paraventricular and supraoptic nuclei in the hypothalamus, and in the nerve cells of the spinal and autonomic ganglions. No PA immunoreactivity was seen in the nucleus and nerve terminals of the neurons. The PA immunoreactivities in the motor and sensory neurons were characterized by clustered masses and blocks of immunoreactive cells. Irrespective of location, small and medium-sized neurons were weakly PA-positive. The glia cells, some stellite cells, and Schwann cells were almost completely PA-negative. These results may suggest that in neurons PAs are not transported axonally, but are located in conjunction with Nissl bodies (the rough endoplasmic reticulum), specified as sites for protein synthesis within cells. PMID:9365032

Fujiwara, K; Bai, G; Kitagawa, T

1997-08-29

 
 
 
 
161

Effects of Static Magnetic Field on Growth of Leptospire, Leptospira interrogans serovar canicola: Immunoreactivity and Cell Division  

CERN Document Server

The effects of the exposure of the bacterium, Leptospira interrogans serovar canicola to a constant magnetic field with magnetic flux density from a permanent ferrite magnet = 140 mT were studied. Changes in Leptospira cells after their exposure to the field were determined on the basis of changes in their growth behavior and agglutination immunoreactivity with a homologous antiserum using darkfield microscopy together with visual imaging. The data showed that the exposed Leptospira cells have lower densities and lower agglutination immunoreactivity than the unexposed control group. Interestingly, some of the exposed Leptospira cells showed abnormal morphologies such as large lengths. We discussed some of the possible reasons for these observations.

Triampo, W; Triampo, D; Wong-Ekkabut, J; Tang, I M; Triampo, Wannapong; Doungchawee, Galayanee; Triampo, Darapond; Wong-Ekkabut, Jirasak

2004-01-01

162

Human lymphocyte production of immunoreactive thyrotropin.  

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Interferon-alpha inducers were previously shown to cause human lymphocyte production of a corticotropin (ACTH)-like peptide. Thyrotropin (TSH) was not produced under these conditions. In contrast, this report shows that a T-cell mitogen (staphylococcal enterotoxin A), which does not induce the ACTH-like peptide, caused human lymphocyte production of an immunoreactive (ir) TSH. Lymphocyte synthesis of the ir TSH was first detectable at 24 hr, peaked at 48 hr, and thereafter declined. NaDodSO4/...

Smith, E. M.; Phan, M.; Kruger, T. E.; Coppenhaver, D. H.; Blalock, J. E.

1983-01-01

163

DCLK1 immunoreactivity in colorectal neoplasia  

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Giuseppe Gagliardi1, Monica Goswami1, Roberto Passera2, Charles F Bellows11Department of Surgery and Pathology, Tulane University, New Orleans, LA, USA; 2Division of Nuclear Medicine Azienda Ospedaliero-Universitaria San Giovanni Battista, Turin, ItalyIntroduction: Microtubule-associated doublecortin and CaM kinase-like-1 (DCLK1) is a novel candidate marker for intestinal stem cells. The aim of our study was to assess DCLK1 immunoreactivity in colorectal carcinogenesis and its correlation wit...

2012-01-01

164

Immunoreactive dynorphin in pituitary and brain.  

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Distribution of the potent opioid peptide dynorphin has been determined in pituitary gland (pig, beef, rat), in the various regions of rat brain, and in rat spinal cord, by using a highly specific antiserum. By gel permeation chromatography in 4 M guanidine, the porcine pituitary immunoreactivity is found in a major peak of apparent molecular weight about 1700 and a minor peak of about 3400. Similar peaks are found in rat pituitary extracts, whereas rat brain contains, in addition, two peaks ...

Goldstein, A.; Ghazarossian, V. E.

1980-01-01

165

Pericellular Innervation of Neurons Expressing Abnormally Hyperphosphorylated Tau in the Hippocampal Formation of Alzheimer's Disease Patients  

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Neurofibrillary tangles (NFT) represent one of the main neuropathological features in the cerebral cortex associated with Alzheimer's disease (AD). This neurofibrillary lesion involves the accumulation of abnormally hyperphosphorylated or abnormally phosphorylated microtubule-associated protein tau into paired helical filaments (PHF-tau) within neurons. We have used immunocytochemical techniques and confocal microscopy reconstructions to examine the distribution of PHF-tau-immunoreactive (ir)...

Blazquez-llorca, Lidia; Garcia-marin, Virginia; Felipe, Javier

2010-01-01

166

Perineurium talin immunoreactivity decreases in diabetic neuropathy.  

Science.gov (United States)

We studied the immunolocalization of Dp116 (a 116 kDa protein product of the dystrophin gene), vinculin, talin, vimentin, desmin, spectrin and titin in the sural nerve biopsies of 25 patients with peripheral neuropathies of different origin. 4 patients presented with HMSN type 1, 4 with HMSN type 2, 2 with HNPP, 4 with CIDP, 5 with chronic axonal neuropathy of unknown origin, 3 with vasculitic neuropathy, 3 with diabetic neuropathy. Expression and localization of Dp116, vinculin, vimentin, desmin, spectrin and titin did not differ from normal control cases. Spectrin and titin immunoreactivities were absent and desmin was occasionally found in few epineurial vessels. A thin rim of Dp116 binding surrounded the outermost layer of myelin sheaths. Perineurium and epineurial vessels stained deeply for vinculin. Vimentin immunoreactivity was seen in all endoneurial, perineurial and epineurial cells. Immunoreactivity for talin was normally found at endoneurial and epineurial vessel walls, perineurial cells and epineurial fibroblasts in all the sural nerves except diabetic nerves. In the latter, whereas talin binding was normal in the vessel walls and epineurial fibroblasts, it was markedly reduced in the perineurium. On immunoblot, two bands at 235 and 190 kDa were found in the sural nerves with the antibody anti-talin, and both were reduced only in the patients with diabetic neuropathy. We postulate that decreased perineurium talin in diabetic polyneuropathy may be related to the known alterations of the tight junctions of the perineurial cells, which have been proposed to be a contributory factor to impaired permeability barrier properties. PMID:9077489

Mazzeo, A; Rodolico, C; Monici, M C; Migliorato, A; Aguennouz, M; Vita, G

1997-02-27

167

Methods for concentration of urinary immunoreactive insulin.  

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Insulin is readily concentrated from 10 to 50 ml of urine with better than 75% recovery using octadecylsilyl (ODS) silica columns (C18Sep-Pak cartridge) and can then be measured by radioimmunoassay. Fractionation on Sephadex G50 gel filtration reveals that the apparent immunoreactivity corresponds for the most part to 6000 dalton insulin. Renal clearance of insulin in 5 normal subjects does not appear to differ in the fasted or fed state and ranged from 0.34 to 0.58 ml/min with an average of ...

Du, B. H.; Eng, J.; Yalow, R. S.

1986-01-01

168

Immunoreactivity assay for ?-particle emitting monoclonal antibody constructs  

International Nuclear Information System (INIS)

Clinical trials using ?-particle radiolabeled antibodies require a rapid and reproducible assay of the immunoreactivity of drugs. While live cell assays are typically used to determine the immunoreactive drug fraction, a fixed cell assay may replace the traditional live cell assay and offer the advantages of rapidity, easy availability and consistency for qualifying drugs for preclinical or clinical studies. We have identified optimal cell fixation and immunoreactivity assay conditions and have validated them by performing the fixed-cell assay in clinical trials

2006-04-01

169

Immunoreactive trypsin and neonatalscreening for cystic fibrosis  

International Nuclear Information System (INIS)

Immunoreactive trypsin (IRT) was measured in dried blood spots from 160.822 five-day-old babies as a part of a regionwide neonatal screening program for cystic fibrosis. A second test was performed for 492 babies in whom blood IRT levels were found greater than 900 ?g/l; retesting revealed persistent elevation in 55. Sweat testing confirmed cystic fibrosis in 43 babies, but results were normal in 12. During the course of this study, a total of 51 cystic fibrosis babies were identified: 43 by newborn screening, 6 because they had meconium ileus; so, early diagnosis was achieved in 49 cases out of 51. Two newborn babies did not have elevated IRT and they were missed by the screening test. Our results confirm that elevated blood IRT is characteristic of newborn babies with cystic fibrosis and show that this test has an excellent specificity (99.7%) and a good sensitivity (95%) when used as a neonatal screening test

1988-01-01

170

Immunoreactive trypsin and neonatalscreening for cystic fibrosis  

Energy Technology Data Exchange (ETDEWEB)

Immunoreactive trypsin (IRT) was measured in dried blood spots from 160.822 five-day-old babies as a part of a regionwide neonatal screening program for cystic fibrosis. A second test was performed for 492 babies in whom blood IRT levels were found greater than 900 ..mu..g/l; retesting revealed persistent elevation in 55. Sweat testing confirmed cystic fibrosis in 43 babies, but results were normal in 12. During the course of this study, a total of 51 cystic fibrosis babies were identified: 43 by newborn screening, 6 because they had meconium ileus; so, early diagnosis was achieved in 49 cases out of 51. Two newborn babies did not have elevated IRT and they were missed by the screening test. Our results confirm that elevated blood IRT is characteristic of newborn babies with cystic fibrosis and show that this test has an excellent specificity (99.7%) and a good sensitivity (95%) when used as a neonatal screening test.

Travert, G.; Laroche, D.; Blandin, C.; Pasquet, C.

1988-01-01

171

FMRFamide-like immunoreactivity in the nervous system of Hydra  

DEFF Research Database (Denmark)

FMRFamide-like immunoreactivity has been localized in different parts of the hydra nervous system. Immunoreactivity occurs in nerve perikarya and processes in the ectoderm of the lower peduncle region near the basal disk, in the ectoderm of the hypostome and in the ectoderm of the tentacles. The immunoreactive nerve perikarya in the lower peduncle region form ganglion-like structures. Radioimmunoassays of extracts of hydra gave displacement curves parallel to standard FMRFamide and values of at least 8 pmol/gram wet weight of FMRFamide-like immunoreactivity. The immunoreactive material eluted from Sephadex G-50 in several components emerging shortly before or after position of authentic FMRFamide. The presence of FMRFamide-like material in coelenterates shows that this family of peptides is of great antiquity.

Grimmelikhuijzen, C J; Dockray, G J

1982-01-01

172

AMYGDALOID KINDLING INCREASES ENKEPHALIN-LIKE IMMUNOREACTIVITY BUT DECREASES DYNORPHIN-A-LIKE IMMUNOREACTIVITY IN RAT HIPPOCAMPUS  

Science.gov (United States)

The effects of amygdaloid kindling on the regional levels and distribution of enkephalin-like immunoreactivity (ELI) and dynorphin A-like immunoreactivity (DNLI) were examined. One day after completion of kindling, radioimmunoassay revealed that there was a 70 percent decrease in...

173

TDP-43 mediates degeneration in a novel Drosophila model of disease caused by mutations in VCP/p97  

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Inclusion body myopathy associated with Paget's disease of bone and frontotemporal dementia (IBMPFD) is a dominantly inherited degenerative disorder caused by mutations in the valosin-containing protein (VCP7) gene. VCP (p97 in mouse, TER94 in Drosophila melanogaster, and CDC48 in Saccharomyces cerevisiae) is a highly conserved AAA(+) (ATPases associated with multiple cellular activities) ATPase that regulates a wide array of cellular processes. The mechanism of IBMPFD pathogenesis is unknown...

Ritson, G. P.; Custer, S. K.; Freibaum, B. D.; Guinto, J. B.; Geffel, D.; Moore, J.; Tang, W.; Winton, M. J.; Neumann, M.; Trojanowski, J. Q.; Lee, V. M. Y.; Forman, M. S.; Taylor, J. P.

2010-01-01

174

Diffusion tensor imaging analysis of sequential spreading of disease in amyotrophic lateral sclerosis confirms patterns of TDP-43 pathology.  

Science.gov (United States)

Diffusion tensor imaging can identify amyotrophic lateral sclerosis-associated patterns of brain alterations at the group level. Recently, a neuropathological staging system for amyotrophic lateral sclerosis has shown that amyotrophic lateral sclerosis may disseminate in a sequential regional pattern during four disease stages. The objective of the present study was to apply a new methodological diffusion tensor imaging-based approach to automatically analyse in vivo the fibre tracts that are prone to be involved at each neuropathological stage of amyotrophic lateral sclerosis. Two data samples, consisting of 130 diffusion tensor imaging data sets acquired at 1.5 T from 78 patients with amyotrophic lateral sclerosis and 52 control subjects; and 55 diffusion-tensor imaging data sets at 3.0 T from 33 patients with amyotrophic lateral sclerosis and 22 control subjects, were analysed by a tract of interest-based fibre tracking approach to analyse five tracts that become involved during the course of amyotrophic lateral sclerosis: the corticospinal tract (stage 1); the corticorubral and the corticopontine tracts (stage 2); the corticostriatal pathway (stage 3); the proximal portion of the perforant path (stage 4); and two reference pathways. The statistical analyses of tracts of interest showed differences between patients with amyotrophic lateral sclerosis and control subjects for all tracts. The significance level of the comparisons at the group level was lower, the higher the disease stage with corresponding involved fibre tracts. Both the clinical phenotype as assessed by the amyotrophic lateral sclerosis functional rating scale-revised and disease duration correlated significantly with the resulting staging scheme. In summary, the tract of interest-based technique allowed for individual analysis of predefined tract structures, thus making it possible to image in vivo the disease stages in amyotrophic lateral sclerosis. This approach can be used not only for individual clinical work-up purposes, but enlarges the spectrum of potential non-invasive surrogate markers as a neuroimaging-based read-out for amyotrophic lateral sclerosis studies within a clinical context. PMID:24736303

Kassubek, Jan; Müller, Hans-Peter; Del Tredici, Kelly; Brettschneider, Johannes; Pinkhardt, Elmar H; Lulé, Dorothée; Böhm, Sarah; Braak, Heiko; Ludolph, Albert C

2014-06-01

175

Ataxin-2 intermediate-length polyglutamine expansions are associated with increased risk for ALS  

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Amyotrophic lateral sclerosis (ALS) is a devastating human neurodegenerative disease. The causes of ALS are poorly understood, although the protein TDP-43 has been suggested to play a critical role in disease pathogenesis. Here we show that Ataxin-2, a polyglutamine (polyQ) protein mutated in spinocerebellar ataxia type 2 (SCA2), is a potent modifier of TDP-43 toxicity in animal and cellular models. The proteins associate in a complex that depends on RNA. Ataxin-2 is abnormally localized in s...

Elden, Andrew C.; Kim, Hyung-jun; Hart, Michael P.; Chen-plotkin, Alice S.; Johnson, Brian S.; Fang, Xiaodong; Armakola, Maria; Geser, Felix; Greene, Robert; Lu, Min Min; Padmanabhan, Arun; Clay, Dana; Mccluskey, Leo; Elman, Lauren; Juhr, Denise

2010-01-01

176

Serum immunoreactive calcitonin concentration in hepatocellular carcinoma  

International Nuclear Information System (INIS)

Having found raised serum calcitonin concentrations is 94% of patients with hepatocellular carcinoma when using a dextran-coated-charcoal radio-immunoassay, we have now repeated the study, using a double-antibody radio-immunoassay, in 102 further patients with hepatocellular carcinoma and 35 matched controls. Serum immunoreactive calcitonin concentrations (iCT) in the controls ranged from 10 to 310 pg/ml (mean 154,6 pg/ml). Values in the tumour patients ranged from 10 to 1 650 pg/ml (mean 302,6 pg/ml). The mean figures were significantly higher in the tumour patients (P smaller than 0,001), 35,5% of them having values above 310 pg/ml. In 65 of the patients serum iCT concentrations were also determined by dextran-coated-charcoal radio-immunoassay. Values ranged from 10 to 10780 pg/ml (mean 2 179 pg/ml). If 1 000 pg/ml is taken as the upper limit of normal, 69% of the patients had raised iCT concentrations. There was a good correlation (r=0,67; P smaller than 0,001) between serum iCT values measured with both methods in 50 patients. If measured by the double-antibody radio-immunoassay method, the serum calcitonin value is not useful as a marker for hepatocellular carcinoma

1982-08-21

177

DCLK1 immunoreactivity in colorectal neoplasia  

Directory of Open Access Journals (Sweden)

Full Text Available Giuseppe Gagliardi1, Monica Goswami1, Roberto Passera2, Charles F Bellows11Department of Surgery and Pathology, Tulane University, New Orleans, LA, USA; 2Division of Nuclear Medicine Azienda Ospedaliero-Universitaria San Giovanni Battista, Turin, ItalyIntroduction: Microtubule-associated doublecortin and CaM kinase-like-1 (DCLK1 is a novel candidate marker for intestinal stem cells. The aim of our study was to assess DCLK1 immunoreactivity in colorectal carcinogenesis and its correlation with prognosis.Methods: DCLK1 immunostaining was performed in colorectal tissue from 71 patients, including 18 adenomatous polyps, 40 primary adenocarcinomas, and 14 metastatic lesions. Each case was evaluated by a combined scoring method based on the intensity of staining (score 0–3 and the percentage of tissue staining positive (score 0–3. Immunoexpression for DCLK1 was considered as positive when the combined score was 2–6 and negative with a score of 0–1.Results: Overall, 14/18 (78% of polyps, 30/40 (75% of primary adenocarcinomas, and 7/14 (50% of distant metastases were positive for DCLK1. In adenomatous polyps and primary cancer there was no association between DCLK1 staining score and tumor pathology. However, after curative colorectal cancer resection, patients whose tumor had a high (?5 combined staining score had increased cancer-specific mortality compared to patients with low (0–4 staining score (hazard ratio 5.89; 95% confidence interval: 1.22–28.47; P = 0.027.Conclusion: We found that DCLK1 is frequently expressed in colorectal neoplasia and may be associated with poor prognosis. Further studies are necessary to validate the use of DCLK1 as a prognostic marker.Keywords: DCLK1, DCAMKL-1, gastrointestinal stem cell, cancer stem cell, adenomatous polyps, liver metastasis, immunohistochemistry

Bellows CF

2012-04-01

178

Growth-hormone-releasing factor immunoreactivity in human endocrine tumors.  

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Seventy-three human tumors and adjacent nonneoplastic tissues were analyzed immunohistochemically for the presence of growth-hormone-releasing factor (GRF). Four of 9 pancreatic endocrine tumors, 2 of 3 appendiceal carcinoids, and 1 of 5 cecal carcinoids were immunoreactive for GRF. One of the GRF-containing pancreatic tumors was associated with acromegaly. Histologically, the growth patterns of these tumors were variable, and the distribution of immunoreactive cells was patchy and irregular....

Bostwick, D. G.; Quan, R.; Hoffman, A. R.; Webber, R. J.; Chang, J. K.; Bensch, K. G.

1984-01-01

179

Isolation and Characterization of Immunoreactive Somatostatin from Fish Pancreatic Islets  

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Using a radioimmunoassay with labeled synthetic tetradecapeptide somatostatin, a large amount of immunoreactive somatostatin was found in the principal pancreatic islet of the channel catfish (Ictalurus punctata). The purpose of these experiments was to isolate and characterize the somatostatin-like material. Extracts of islets were chromatographed on a Bio-Gel P-30 column, and over 90% of the immunoreactive somatostatin migrated with proteins at least twice the size of synthetic tetradecapep...

1980-01-01

180

Temperature-dependent immunoreactive assay to screen for digoxin-like immunoreactive factor(s).  

Science.gov (United States)

Endogenous circulating digoxin-like immunoreactive factors (DLIF) are known to cross-react with antibodies to digoxin and to inhibit Na+/K(+)-transporting ATPase (Na+K+ATPase; EC 3.6.1.37). Moreover, increasing the immunoassay temperature from 4 to 37 degrees C markedly decreases DLIF from human cord serum. We tested several compounds, including hormonal steroids, bile salts, lipids, and methionine-enkephalin, for their ability to cross-react with two commercially available 125I digoxin RIAs, to inhibit porcine Na+K+ATPase, and to see whether they present the same incubation temperature dependence as human cord serum. Except for methionine-enkephalin, all compounds were inhibitors of Na+K+ATPase in the range of 1-10 mmol/L. Progesterone exhibited the highest cross-reactivity in the two RIAs. The apparent digoxin immunoreactivity for the majority of the cross-reacting steroids, bile salts, and linoleic acid was markedly decreased by increasing the incubation temperature from 4 to 37 degrees C, whereas estriol, pregnanediol, and nonspecific compounds (e.g., ethanol, human serum albumin) did not appear to be temperature-sensitive. Both lysophosphatidyl lipids gave an increased apparent digoxin concentration with increasing incubation temperature. Our data suggest that numerous weakly cross-reactive compounds can parallel the response of human cord serum. However, the temperature-dependent effect could be an additional criterion for identifying DLIF. PMID:1718632

Guédeney, X; Chanez, C; Grenier, A; Scherrmann, J M

1991-11-01

 
 
 
 
181

Skeletal limb abnormalities  

Science.gov (United States)

Skeletal limb abnormalities refer to a variety of bone structure problems in the arms or legs (limbs). ... Skeletal limb abnormalities are most often used to describe defects in the legs or arms that are ...

182

Ataxin-2 intermediate-length polyglutamine expansions are associated with increased risk for ALS.  

Science.gov (United States)

The causes of amyotrophic lateral sclerosis (ALS), a devastating human neurodegenerative disease, are poorly understood, although the protein TDP-43 has been suggested to have a critical role in disease pathogenesis. Here we show that ataxin 2 (ATXN2), a polyglutamine (polyQ) protein mutated in spinocerebellar ataxia type 2, is a potent modifier of TDP-43 toxicity in animal and cellular models. ATXN2 and TDP-43 associate in a complex that depends on RNA. In spinal cord neurons of ALS patients, ATXN2 is abnormally localized; likewise, TDP-43 shows mislocalization in spinocerebellar ataxia type 2. To assess the involvement of ATXN2 in ALS, we analysed the length of the polyQ repeat in the ATXN2 gene in 915 ALS patients. We found that intermediate-length polyQ expansions (27-33 glutamines) in ATXN2 were significantly associated with ALS. These data establish ATXN2 as a relatively common ALS susceptibility gene. Furthermore, these findings indicate that the TDP-43-ATXN2 interaction may be a promising target for therapeutic intervention in ALS and other TDP-43 proteinopathies. PMID:20740007

Elden, Andrew C; Kim, Hyung-Jun; Hart, Michael P; Chen-Plotkin, Alice S; Johnson, Brian S; Fang, Xiaodong; Armakola, Maria; Geser, Felix; Greene, Robert; Lu, Min Min; Padmanabhan, Arun; Clay-Falcone, Dana; McCluskey, Leo; Elman, Lauren; Juhr, Denise; Gruber, Peter J; Rüb, Udo; Auburger, Georg; Trojanowski, John Q; Lee, Virginia M-Y; Van Deerlin, Vivianna M; Bonini, Nancy M; Gitler, Aaron D

2010-08-26

183

Novel neuritic clusters with accumulations of amyloid precursor protein and amyloid precursor-like protein 2 immunoreactivity in brain regions damaged by thiamine deficiency.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Experimental thiamine deficiency (TD) is a classical model of a nutritional deficit associated with a generalized impairment of oxidative metabolism and selective cell loss in the brain. In rats, TD-induced cell degeneration is accompanied by an accumulation of amyloid precursor protein (APP)/amyloid precursor-like protein 2 (APLP2) immunoreactivity in abnormal neurites and perikarya along the periphery of, or scattered within, the lesion. Prompted by these data and our previous findings of a...

Calingasan, N. Y.; Gandy, S. E.; Baker, H.; Sheu, K. F.; Smith, J. D.; Lamb, B. T.; Gearhart, J. D.; Buxbaum, J. D.; Harper, C.; Selkoe, D. J.; Price, D. L.; Sisodia, S. S.; Gibson, G. E.

1996-01-01

184

Urine - abnormal color  

Science.gov (United States)

The usual color of urine is straw-yellow. Abnormally colored urine may be cloudy, dark, or blood-colored. ... Abnormal urine color may be caused by infection, disease, medicines, or food you eat. Cloudy or milky urine is a sign ...

185

Normal Merkel cells express a synaptophysin-like immunoreactivity.  

Science.gov (United States)

Synaptophysin (SY), a specific component of the membrane of presynaptic vesicles, has been reported as a novel marker for neurons, certain neuroendocrine cells and their neoplasms including neuroendocrine carcinomas of the skin. The origin of the Merkel cells (MC) being far from clear, this study was performed to establish if normal MC express SY. It is demonstrated by immunofluorescence and immunoelectron microscopy using a monoclonal antibody SY38 to this glycoprotein that normal MC in man, rabbit and pigs express an SY-like reactivity. Although immunoblotting identification of the immunoreactive material gave negative results, it is likely that normal MC contain SY. By immunoelectron microscopy, the staining was located at the surface of cytoplasmic vesicles. In view of the possible involvement of SY in the Ca2+-dependent neurotransmitter release, the observation of an SY-like immunoreactivity in MC supports the view that they are epithelial neuroendocrine cells and that they may possess a neurosecretory function. PMID:3141225

Ortonne, J P; Petchot-Bacque, J P; Verrando, P; Pisani, A; Pautrat, G; Bernerd, F

1988-01-01

186

Insulin-like immunoreactive substances in the rat  

International Nuclear Information System (INIS)

Chromatography on G50 or G100 sephadex column of rat plasma or serum divides up the insulin-like immunoreactive material into three peaks: monomere insulin, proinsulin and a fraction of molecular weight between 50 and 100,000. This fraction is virtually absent (less than 1%) from immunoreactive material extracted from the pancreas. Comparison of the results obtained by methods using double or simple antibodies (charcoal dextran) and study of fixation in vitro of labelled insulin, taken up by various plasma proteins, suggest that the high molecular weight material includes insulin more or less broken down and linked to proteins. Furthermore, when a double antibody method is used, the alpha globulins and albumin in the rat present also an insulin-like reactivity. This disadvantage does not occur with the charcoal dextran method which is more specific

1975-12-01

187

Distribution and chromatographic analysis of galanin immunoreactivity in the heart.  

Science.gov (United States)

The presence and distribution of the biologically active neuropeptide galanin (GAL), in the rat heart as well as in mouse, guinea pig, rabbit, cat, and dog heart, were analyzed. With some minor variations in the overall distribution, extractable GAL-like immunoreactivity (-LI) was present in all major portions of the heart. In the rat heart, GAL-immunoreactive (GAL-IR) nerve fibers were present in the atria as well as in the ventricles; thin GAL-IR fibers were present in the myocardium as well as around some cardiac blood vessels. A few larger GAL-IR nerve fiber bundles were also present on the surface of the heart. Characterization of extractable GAL-LI in the rat heart, using HPLC, revealed one GAL-IR form, coeluting with synthetic rat GAL. Our findings suggest that galanin is of importance in the control of certain cardiac functions and/or of circulation. PMID:7536325

Xu, Y; Johansson, O; Rökaeus, A

1995-01-01

188

Kinetics of Antibody Response to Ehrlichia canis Immunoreactive Proteins  

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Immunoreactive proteins of Ehrlichia canis and Ehrlichia chaffeensis that have been characterized include a family of 28-kDa major outer membrane proteins (p28) and two large antigenically divergent surface glycoprotein orthologs. We previously demonstrated that recombinant E. canis p28 and the 140- and 200-kDa glycoproteins gp140 and gp200, respectively, react strongly with serum antibodies from suspect canine ehrlichiosis cases that were positive for E. canis by immunofluorescent antibody t...

2003-01-01

189

Chromosomal Abnormalities and Schizophrenia  

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Schizophrenia is a common and serious psychiatric illness with strong evidence for genetic causation, but no specific loci yet identified. Chromosomal abnormalities associated with schizophrenia may help to understand the genetic complexity of the illness. This paper reviews the evidence for associations between chromosomal abnormalities and schizophrenia and related disorders. The results indicate that 22q11.2 microdeletions detected by fluorescence in-situ hybridization (FISH) are significa...

Bassett, Anne S.; Chow, Eva W. C.; Weksberg, Rosanna

2000-01-01

190

Localization of Neuropeptide Y1 Receptor Immunoreactivity in the Rat Retina and the Synaptic Connectivity of Y1 Immunoreactive Cells  

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Neuropeptide Y (NPY), an inhibitory neuropeptide expressed by a moderately dense population of wide-field amacrine cells in the rat retina, acts through multiple (Y1–y6) G-protein–coupled receptors. This study determined the cellular localization of Y1 receptors and the synaptic connectivity of Y1 processes in the inner plexiform layer (IPL) of the rat retina. Specific Y1 immunoreactivity was localized to horizontal cell bodies in the distal inner nuclear layer and their processes in the ...

D Angelo, Iona; Oh, Su-ja; Chun, Myung-hoon; Brecha, Nicholas C.

2002-01-01

191

Immunoreactivity examination of patients with testicular tumours treated with radiotherapy  

International Nuclear Information System (INIS)

Results of the immunoreactivity study of 72 patients receiving radiotherapy are presented. Tuberculin and DNCB (2,4 dinitrochlorobenzol) reactivity tests were performed before, during and 3 years after the radiation therapy and at the time when metastases appeared. The number of positive reactions decreased slightly in both tuberculin and DNCB groups, though not significantly. Metastatic patients showed a significant decrease of reactivity against DNCB as compared with the results obtained before the treatment. In 5,6% of patients herpes zoster was registered. No other infections occured. It was found that immunosuppression caused by the radiation treatment does not influence the later fate of patients with testicular tumours. (author)

1985-02-01

192

Serotonin immunoreactivity in the carotid body of adult humans.  

Science.gov (United States)

The distribution of serotonin immunoreactivity (-IR) was studied in adult human carotid bodies, obtained at post-mortem, using both the peroxidase-antiperoxidase method on paraffin sections and a double-labelling immunofluorescence on frozen sections. Antibodies against synaptophysin and protein gene product (PGP) 9.5 were used for identification of serotonin-IR cells. Serotonin-IR was demonstrable in the carotid bodies of adult humans and it was coexpressed mostly with synaptophysin or PGP 9.5 in type I cells. Some serotonin immunopositive type I cells were located in close proximity to capillaries. Serotonin-IR was also observed in a few endothelial cells. PMID:8075479

Habeck, J O; Pallot, D J; Kummer, W

1994-04-01

193

The value of immunoreactive lipase in acute pancreatitis.  

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We have evaluated a new agglutination test for serum immunoreactive lipase in 24 patients with abdominal pain and hyperamylasaemia. On admission all 20 patients with acute pancreatitis had a positive lipase test, 3 of the 4 patients who did not have pancreatitis had a negative lipase test. The sensitivity of the lipase test on day 1 is 100%, the specificity 96% and predictive value of a positive test is 95.2% compared to 83% for amylase. A negative test excludes pancreatitis. In addition, the...

Hemingway, D. M.; Johnson, I.; Tuffnell, D. J.; Croton, R. S.

1988-01-01

194

Immunoreactivity examination of patients with testicular tumours treated with radiotherapy  

Energy Technology Data Exchange (ETDEWEB)

Results of the immunoreactivity study of 72 patients receiving radiotherapy are presented. Tuberculin and DNCB (2,4 dinitrochlorobenzol) reactivity tests were performed before, during and 3 years after the radiation therapy and at the time when metastases appeared. The number of positive reactions decreased slightly in both tuberculin and DNCB groups, though not significantly. Metastatic patients showed a significant decrease of reactivity against DNCB as compared with the results obtained before the treatment. In 5,6% of patients herpes zoster was registered. No other infections occured. It was found that immunosuppression caused by the radiation treatment does not influence the later fate of patients with testicular tumours. 41 refs.

Stefanits, K.; Kuhn, E.; Csere, T.

1985-02-01

195

Isolation and characterization of immunoreactive somatostatin from fish pancreatic islets.  

Science.gov (United States)

Using a radioimmunoassay with labeled synthetic tetradecapeptide somatostatin, a large amount of immunoreactive somatostatin was found in the principal pancreatic islet of the channel catfish (Ictalurus punctata). The purpose of these experiments was to isolate and characterize the somatostatin-like material. Extracts of islets were chromatographed on a Bio-Gel P-30 column, and over 90% of the immunoreactive somatostatin migrated with proteins at least twice the size of synthetic tetradecapeptide somatostatin. This fraction was further purified by ion-exchange chromatography on carboxymethyl-cellulose and DEAE-cellulose columns. Two peptides were obtained with identical immunoreactivity, which was approximately 25% that of the synthetic somatostatin. Each peptide was judged to be >95% pure by thin-layer electrophoresis, polyacrylamide gel electrophoresis at pH 8.9, and highpressure liquid chromatography. Further criteria of purity included amino-terminal analysis of fraction IV yielding only aspartic acid. A total of 1.3 mg of fraction II, and 3.8 mg of fraction IV somatostatin-like peptides were obtained from 10 g of fresh frozen islets. Characterization of the two peptides revealed both peptides slightly more acidic than synthetic tetradecapeptide somatostatin. Fraction II had an isoelectric point of 8.0-8.3, and fraction IV 8.3-9.0. Molecular weight estimation by sodium dodecyl sulfate-urea polyacrylamide gel electrophoresis revealed similar mobility of both peptides, between pancreatic polypeptide (mol wt 4,500) and glucagon (mol wt 3,500). The mobility was not altered by reduction, and was approximately twice the size of synthetic tetradecapeptide somatostatin (mol wt 1,800). This confirmed that the peptides were single polypeptide chains and not aggregates, or somatostatin bound to larger proteins. Molecular weight determination by gel filtration chromatography on Bio-Gel P-6 in 8 M urea gave an estimated mol wt of 3,700. Amino acid analysis of the two immunoreactive somatostatins indicated that they were very similar in composition. Both pancreatic somatostatins (1 muM) had full biological activity relative to synthetic somatostatin measured as inhibition of growth hormone release from rat anterior pituitary cells. PMID:6102573

Oyama, H; Hirsch, H J; Gabbay, K H; Permutt, A

1980-05-01

196

Prognostic importance of proliferating cell nuclear antigen immunoreactivity and mitotic index in malignant mesothelioma  

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Full Text Available Aim: In this study, proliferating nuclear cell antigen (PCNA immunoreactivity and the mitotix index were searched in human malignant mesothelioma to assess their prognostic value.Material and Methods: PCNA immunoreactivity was investigated in 19 cases. The authors also compared this with mitosis counts.Results: There was no correlation between the percentage of PCNA immunoreactive cells and their mitotic counts. However, the median survival was 16.4 months for patients with less than 25% PCNA immunoreactive cells, 17.8 months for patients with less than 4 mitotic figures 10 high power fields of tumoral tissue, 10.8 months for patients with more than 25 per cent PCNA immunoreactive cells, and 14.2 months for patients with more than 4 mitotic figures in 10 high power fields of tumoral tissue.Conclusion: Our results suggest that PCNA immunoreactivity and mitotic count may have prognostic values in malignant mesothelioma.

I??n SOYUER

2002-09-01

197

CT of pleural abnormalities  

International Nuclear Information System (INIS)

Briefly discussed were CT diagnosis of pleural thickening, CT technique for examining the pleura or pleuro-pulmonary disease, diagnosis of pleural collections, diagnosis of pleural fluid abnormalities in patients with pneumonia, pleural neoplasms, malignant (diffuse) mesothelioma, metastases, local fibrous tumor of the pleura (benign mesothelioma) (21 refs.)

1995-06-01

198

Abnormality, rationality, and sanity.  

Science.gov (United States)

A growing body of studies suggests that neurological and mental abnormalities foster conformity to norms of rationality that are widely endorsed in economics and psychology, whereas normality stands in the way of rationality thus defined. Here, we outline the main findings of these studies, discuss their implications for experimental design, and consider how 'sane' some benchmarks of rationality really are. PMID:24055170

Hertwig, Ralph; Volz, Kirsten G

2013-11-01

199

Abnormalities of gonadal differentiation.  

Science.gov (United States)

Gonadal differentiation involves a complex interplay of developmental pathways. The sex determining region Y (SRY) gene plays a key role in testis determination, but its interaction with other genes is less well understood. Abnormalities of gonadal differentiation result in a range of clinical problems. 46,XY complete gonadal dysgenesis is defined by an absence of testis determination. Subjects have female external genitalia and come to clinical attention because of delayed puberty. Individuals with 46,XY partial gonadal dysgenesis usually present in the newborn period for the valuation of ambiguous genitalia. Gonadal histology always shows an abnormality of seminiferous tubule formation. A diagnosis of 46,XY true hermaphroditism is made if the gonads contain well-formed testicular and ovarian elements. Despite the pivotal role of the SRY gene in testis development, mutations of SRY are unusual in subjects with a 46,XY karyotype and abnormal gonadal development. 46,XX maleness is defined by testis determination in an individual with a 46,XX karyotype. Most affected individuals have a phenotype similar to that of Klinefelter syndrome. In contrast, subjects with 46,XX true hermaphroditism usually present with ambiguous genitalia. The majority of subjects with 46,XX maleness have Y sequences including SRY in genomic DNA. However, only rare subjects with 46,XX true hermaphroditism have translocated sequences encoding SRY. Mosaicism and chimaerism involving the Y chromosome can also be associated with abnormal gonadal development. However, the vast majority of subjects with 45,X/46,XY mosaicism have normal testes and normal male external genitalia. PMID:9890065

Berkovitz, G D; Seeherunvong, T

1998-04-01

200

Immunoreactive anionic and cationic trypsin in human serum.  

Science.gov (United States)

A simple method for the purification of anionic and cationic trypsinogen and trypsin from human pancreatic juice applying affinity chromatography on aprotinin coupled Sepharose is described together with the N-terminal amino acid sequences for both trypsinogens. In addition, enzyme-linked immunoabsorbent assay (ELISA) methods for the determination of anionic and cationic trypsin-like immunoreactivity (irAT and irCT) are described. Normal serum levels are 21.3 +/- 7.4 micrograms/l and 27.8 +/- 9.0 microgram/l for irAT and irCT respectively and the accuracy of these assays is 6-10%. In our population, the normal ratio between irCT and irAT in serum is 1.36 +/- 0.42. In normal serum trypsin-like immunoreactivity consists solely of trypsinogen. In acute pancreatitis there is an increase over normal of both irAT and irCT with a proportionally greater increase in irAT than irCT. Similar changes are also found in uremic patients. PMID:2598466

Kimland, M; Russick, C; Marks, W H; Borgström, A

1989-09-15

 
 
 
 
201

Liver abnormalities in pregnancy.  

Science.gov (United States)

Abnormalities of liver function (notably rise in alkaline phosphatase and fall in serum albumin) are common in normal pregnancy, whereas rise in serum bilirubin and aminotransferase suggest either exacerbation of underlying pre-existing liver disease, liver disease related to pregnancy or liver disease unrelated to pregnancy. Pregnant women appear to have a worse outcome when infected with Hepatitis E virus. Liver diseases associated with pregnancy include abnormalities associated hyperemesis gravidarum, acute fatty liver disease, pre-eclampsia, cholestasis of pregnancy and HELLP syndrome. Prompt investigation and diagnosis is important in ensuring a successful maternal and foetal outcome. In general, prompt delivery is the treatment of choice for acute fatty liver, pre-eclampsia and HELLP syndrome and ursodeoxycholic acid is used for cholestasis of pregnancy although it is not licenced for this indication. PMID:24090943

Than, Nwe Ni; Neuberger, James

2013-08-01

202

Neurodevelopmental Abnormalities in ADHD  

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Structural and functional imaging studies in subjects with attention deficit hyperactivity disorder (ADHD) are reviewed with the goal of gleaning information about neurodevelopmental abnormalities characterizing the disorder. Structural imaging studies, particularly those with longitudinal designs, suggest that brain maturation is delayed by a few years in ADHD. However, a maturational delay model alone is incomplete: alternate courses are suggested by differences associated with phenotypic f...

Vaidya, Chandan J.

2012-01-01

203

Plant abnormality detection device  

International Nuclear Information System (INIS)

When the result of judgement recognized by an operator is different from the result of judgement reported by the system, for example, in a power plant, the system of the present invention can easily compensate it by manual input at a level of threshold value. Namely, when abnormality of rotational equipments of a plant is monitored by a microphone, acoustic signals caught by the microphone are converted to a sound pressure level. Degree of abnormality confidence is calculated in order to compare this with a threshold value. On the other hand, in order to determine the threshold value, information to be provided to an operator is calculated by obtaining data from a distribution data calculation portion is calculated at a compensation section. The threshold value level is displayed while being overlaid on a probability density distribution graph on a CRT. An operator compensates the level of the threshold value while observing the CRT screen. In order to compare the threshold level with the degree of abnormality confidence, a threshold value exceeding probability calculation section calculates the probability of occurrence relative to probability density distribution. (I.S.)

1994-07-29

204

Responses of plasma cyclic AMP, serum immunoreactive insulin, C-peptide immunoreactivity and blood sugar levels to glucagon in patients with liver diseases.  

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Levels of plasma cyclic AMP, serum immunoreactive insulin (IRI), serum c-peptide immunoreactivity (CPR) and blood sugar (BS) were determined 0, 15, 30, 45 and 60 min after a glucagon injection (0.01 mg per kg body weight) in normal controls, patients with acute hepatitis and liver cirrhosis. Plasma cyclic AMP responses to glucagon in liver disease patients varied widely in peak value, and only in patients with fulminant hepatitis and decompensated liver cirrhosis with poor prognosis was th...

Shimamura, Junnosuke; Taketa, Kazuhisa; Ide, Takero; Nakata, Kenichi; Nagashima, Hideo

1985-01-01

205

Immunoreactive atrial natriuretic peptide in the guinea pig spleen  

International Nuclear Information System (INIS)

The presence of immunoreative ANP precursor-like material in the guinea pig spleen is suggested. This is based on the following experimental evidence: An acidic extract of guinea pig spleen analyzed by Sephadex G-50 gel filtration contained 4.6 pmol/g wet tissue immunoreactive atrial natriuretic peptide (IR-ANP), IR-ANP coeluting with 15 kDa synthetic ANP (2-126). Gel filtrated IR-ANP material was further submitted to reverse phase high performance liquid chromatography and monitored by radioimmunoassay employing two antisera. One antiserum recognizes the C-terminal of ANP (1-126), the second is directed against the N-terminal sequence. Both antisera revealed material eluting with synthetic ANP (2-126). Furthermore, immunohistochemical analysis suggests this ANP-like material to be localized mainly at the periphery of the white pulp of the spleen. These findings link ANP with the immune system

1989-01-01

206

Epilepsy and chromosomal abnormalities  

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Full Text Available Abstract Background Many chromosomal abnormalities are associated with Central Nervous System (CNS malformations and other neurological alterations, among which seizures and epilepsy. Some of these show a peculiar epileptic and EEG pattern. We describe some epileptic syndromes frequently reported in chromosomal disorders. Methods Detailed clinical assessment, electrophysiological studies, survey of the literature. Results In some of these congenital syndromes the clinical presentation and EEG anomalies seems to be quite typical, in others the manifestations appear aspecific and no strictly linked with the chromosomal imbalance. The onset of seizures is often during the neonatal period of the infancy. Conclusions A better characterization of the electro clinical patterns associated with specific chromosomal aberrations could give us a valuable key in the identification of epilepsy susceptibility of some chromosomal loci, using the new advances in molecular cytogenetics techniques - such as fluorescent in situ hybridization (FISH, subtelomeric analysis and CGH (comparative genomic hybridization microarray. However further studies are needed to understand the mechanism of epilepsy associated with chromosomal abnormalities.

Sorge Giovanni

2010-05-01

207

Abnormality analyzing device  

International Nuclear Information System (INIS)

The device of the present invention analyzes a steam generator of a PWR plant easily and accurately. A signal generator sends maximum length sequential signals to an adder, in which M sequence signals having a sufficiently large amplitude than that of noises of correction control signals to provide zero external disturbance in average are added in the adder and outputted. A PI controller receives the signals, conducts PI calculation and sent them as control signals containing disturbance to a feedwater control valve. The feedwater control valve opens/closes depending on the signals and keeps the water level in the same manner as in the case where there is no disturbance, namely, in a case where M sequential signals are not present for long period of time. It stores flow rate signals of a feedwater valve containing external disturbance and water level signals of the steam generator, and calculates a predetermined coefficient of a vector of approximate transmission function of the steam generator. The calculated value and the coefficient of vector of the approximate transmission function during normal operation are compared, and when the difference exceeds a predetermined value, it is determined as abnormal, and abnormal signals are outputted. It is not necessary to obtain detailed transmission function of the steam generator, but analysis can be conducted easily and accurately by the approximate transmission function. (N.H.)

1996-02-16

208

Plasma immunoreactive somatostatin is elevated in diabetic ketoacidosis and correlates with plasma non-esterified fatty acid concentration.  

Science.gov (United States)

In experimental diabetes and after the administration of beta-hydroxybutyrate and non-esterified fatty acids (NEFA), an increase in circulating immunoreactive somatostatin (IRS) has been described. Both ketones and NEFA are raised in diabetic ketoacidosis. Therefore, we decided to investigate 10 patients in diabetic ketoacidosis by measuring, on admission and throughout the initial 24 hours of therapy, circulating levels of IRS, beta-hydroxybutyrate, acetoacetate, triglycerides, blood glucose, pH and NEFA. Fluids and insulin were administered IV following a previously established protocol. Nine patients showed abnormally high levels of circulating IRS. When compared with a group of controlled insulin-dependent diabetic patients, basal IRS was high (111 +/- 15 vs 28 +/- 3 pmol/l), and remained elevated for at least 24 h despite clear improvement of metabolic status. On admission we also found elevated levels of NEFA (1.04 +/- 0.2 mmol/l), triglycerides (4.7 +/- 1.1 mmol/l), beta-hydroxybutyrate (22.1 +/- 4mmol/l), and acetoacetate (4.8 +/- 1.1 mmol/l). A significant correlation was found initially between IRS and NEFA (p less than 0.01). We conclude that circulating IRS is high in most cases of diabetic ketoacidosis. The mechanism behind this hypersomatostatinaemia could be related to the abnormalities of lipid metabolism which occur in diabetic ketoacidosis. PMID:2886248

Binimelis, J; Webb, S M; Serrano, J; Codina, M; Corcoy, R; Perez, A; Peinado, M A; Puig, M; de la Torre, W; Serrat, J

1987-01-01

209

Eye movement abnormalities.  

Science.gov (United States)

Generation and control of eye movements requires the participation of the cortex, basal ganglia, cerebellum and brainstem. The signals of this complex neural network finally converge on the ocular motoneurons of the brainstem. Infarct or hemorrhage at any level of the oculomotor system (though more frequent in the brain-stem) may give rise to a broad spectrum of eye movement abnormalities (EMAs). Consequently, neurologists and particularly stroke neurologists are routinely confronted with EMAs, some of which may be overlooked in the acute stroke setting and others that, when recognized, may have a high localizing value. The most complex EMAs are due to midbrain stroke. Horizontal gaze disorders, some of them manifesting unusual patterns, may occur in pontine stroke. Distinct varieties of nystagmus occur in cerebellar and medullary stroke. This review summarizes the most representative EMAs from the supratentorial level to the brainstem. PMID:22377853

Moncayo, Jorge; Bogousslavsky, Julien

2012-01-01

210

Abnormal Occupation REvisited  

CERN Multimedia

It is demonstrated the Fermi surface of dense neutron matter may experience a rearrangement near the onset of pion condensation, due to strong momentum dependence of the effective interaction induced by spin-isospin fluctuations. In particular, the Fermi surface may take the form of a partially hollow sphere having a spherical hole in its center (a ``Lifschitz bubble''). Thus, a second (inner) Fermi surface may form as high-momentum single-particle states are filled and low-momentum states are vacated. The influence of this phenomenon on the superfluid transition temperature of the Fermi system is characterized with the help of a separation transformation of the BCS gap equation. This work may be viewed as a revival of the search for physical realizations of abnormal occupation in infinite, homogeneous Fermi systems -- plausible instances in which the quasiparticle distribution differs from that of an ideal Fermi gas and Fermi liquid theory breaks down.

Clark, J W; Zverev, M V

2001-01-01

211

Total digoxin-like immunoreactive factor(s) in healthy population, uncomplicated term pregnancies and neonates.  

Science.gov (United States)

Free digoxin-like immunoreactive factor(s) (DLIF) which may have a homeostatic role, as documented in different physiological conditions, but is generally undetectable in plasma from normal population. Total digoxin-like immunoreactive factor(s) (protein bound and free) can be estimated after plasma is heated. In this study, total digoxin-like immunoreactive factor(s) as measured in plasma in a well defined control population and compared to healthy term pregnant women and neonates, categories known to be associated with increased free digoxin-like immunoreactive factor(s) concentrations. The mean level of this factor(s) in the control group was 706 +/- 129 pg digoxin equivalent/ml (pg/ml) and was unaffected by age and sex. Significantly increased levels of total digoxin-like immunoreactive factor(s) were found in pregnant women and neonates (928 +/- 127 and 1242 +/- 367 pg/ml, respectively). We conclude that levels of total digoxin-like immunoreactive factor(s) are increased in term pregnancies and neonates, similarly to its free form. However total digoxin-like immunoreactive factor(s) is detected in the normal population as a plasma component, contrary to its free form, which is generally undetectable. PMID:2319115

Krivoy, N; Jakobi, P; Paldi, E; Alroy, G

1990-01-01

212

Sleep deprivation reduces neuroglobin immunoreactivity in the rat brain.  

Science.gov (United States)

Neuroglobin (Ngb), a protein located in the mammal's brain, is involved in oxygen transport and free radical scavenging inside the neurons. Ngb colocalizes with choline acetyltransferase in the laterodorsal tegmental nucleus and in the pontine tegmental nucleus, both involved in the sleep-wake cycle regulation. Some studies have shown that free radicals accumulated during prolonged wakefulness are removed during sleep. Therefore, Ngb could act as a regulator of free radicals generated during prolonged wakefulness in the brain. The aim of this study was to determine whether prolonged wakefulness affects Ngb immunoreactivity because of increases in the oxidative stress induced by continuous neuronal activity. For this purpose, male adult Wistar rats were implanted with electrodes for sleep recordings and were divided into control and sleep-deprived groups. Sleep deprivation was carried out for 24 h by gentle handling of the animals. Sleep-wake activity was determined during the deprivation period or 24?h of control conditions. Subsequently, both groups of animals were killed and their brains were obtained and processed for Ngb immunohistochemical analysis and detection of lipid peroxidation. Our data found no evidence of increased oxidative stress in the brains of sleep-deprived animals compared with the controls. The number of Ngb-positive cells was decreased in the sleep-deprived animals in all analyzed areas of the brain compared with the control group. Our results suggest that Ngb could be involved in sleep regulation, independent of its role in the control of oxidative stress. PMID:23262504

Melgarejo-Gutiérrez, Montserrat; Acosta-Peña, Eva; Venebra-Muñoz, Arturo; Escobar, Carolina; Santiago-García, Juan; Garcia-Garcia, Fabio

2013-02-13

213

Serum immunoreactive erythropoietin in HIV-infected patients  

International Nuclear Information System (INIS)

Serum immunoreactive erythropoietin (SIE) and hemoglobin levels were measured in 152 patients infected with the human immunodeficiency virus. Anemia was present in 18% of asymptomatic patients who tested positive for the human immunodeficiency virus, 50% of patients with a condition related to the acquired immunodeficiency syndrome (AIDS), and 75% of patients with AIDS. The mean SIE level for untreated AIDS patients was greater than for patients who tested positive for human immunodeficiency virus or patients with an AIDS-related condition but not outside the normal range for SIE, and the incremental increase in SIE level for a given decline in hemoglobin level was much less in AIDS patients than in patients with uncomplicated iron deficiency anemia. Forty-two patients were treated with zidovudine, and the hemoglobin level fell 10 g/L or more in 48%. The data indicate that SIE level is inappropriately low in anemic AIDS patients. The ability of these patients to produce erythropoietin is intact and can be expressed with zidovudine therapy. However, even very high levels of SIE fail to stimulate erythropoiesis adequately

1989-06-02

214

Clinical applications of measurement of serum immunoreactive levels of erythropoietin  

International Nuclear Information System (INIS)

The purification of erythropoietin (Ep) in 1977 enabled investigators to more clearly define the role of this hormone in erythropoiesis in man. Radioimmunoassays were rapidly developed. Undoubtedly differences between levels of immunoreactive and biologically active Ep will be found but the resolution of these discrepancies will expand our understanding of the erythron. Recently others described a monoclonal antibody against Ep. Because of this breakthrough, large quantities of pure hormone should soon be available to a larger number of investigators than currently have access to it. The major clinical use of this hormone will probably be in the treatment of the anemia of chronic renal disease. In the relatively few years since the radioimmunoassay (RIA) was developed, measurements of the levels of this hormone have been made in several disease states as well as in normal man. Most of the findings to date confirm the predictions that have been made over the years based on studies done using the rather crude bioassay for Ep. In the present study the authors shall review and expand on what is known about subjects with chronic lung and renal disease

1985-01-01

215

Deposition of immunoreactants in a cutaneous allergic drug reaction  

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Full Text Available Context: The analysis of allergic drug reaction pathology may be difficult, especially if multiple histological reaction patterns are detected on review of hematoxylin and eosin (H&E stained sections. In this case, we emphasize the value of adding immunohistochemistry (IHC and multicolor direct immunofluorescence (DIF as tools to improve the diagnosis of these complex disorders. Patient and Methods: Our patient is a twenty-year-old Caucasian female, who presented with a sudden onset of erythematous macules on the skin following administration of amoxicillin. Lesional tissue was examined by H & E and IHC, and perilesional tissue by DIF and IHC. Results: The H&E findings revealed diffuse dermal edema, and a mild, superficial, perivascular dermatitis with a mixed inflammatory infiltrate, consistent with an allergic drug eruption. The IHC and DIF studies revealed autoreactivity to sweat glands, nerves and dermal blood vessels, as well as dermal deposits of immune reactants such as fibrinogen and complement around the inflamed areas. Conclusions: Fibrin-fibrinogen degradation products have been shown in some cases of allergic disorders; thus, we encourage the effect further testing for these immunoreactants in biopsies from patients with possible allergic drug reactions.

Ana Maria Abreu Velez

2009-09-01

216

Immunoreactive neuropeptides in the cells of human thymus  

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Full Text Available The study was designed to explore the expression of different neuropeptides, viz. vasoactive intestinal peptide (VIP, calcitonin gene related peptide (CGRP, substance P (SP, bombesin and motilin in the cells of fetal and adult human thymus. Immunohistochemical staining revealed that cortical and medullary thymocytes were labeled by all antibodies, except those specific for motilin. Immunoreactive VIP and SP were observed in the solitary epithelial cells located in the subcapsular/subtrabecular cortex, at the corticomedullary junction and in the medulla. The cells within the subcapsular/subtrabecular monolayer, rare solitary cells in the deep cortex and epithelial cell network in the medulla, were labeled with antibodies to CGRP and bombesin. Hassall’s corpuscles were labeled with all antibodies except that specific for SP. The obtained data obtained testify to the expression of different neuropeptides in human thymic lymphoid and non-lymphoid cells and suggest a role for neuroendocrine hormone-mediated mechanisms in the regulation of thymic homeostasis in humans.

Leposavi? Gordana

2011-01-01

217

Serum immunoreactive erythropoietin in HIV-infected patients  

Energy Technology Data Exchange (ETDEWEB)

Serum immunoreactive erythropoietin (SIE) and hemoglobin levels were measured in 152 patients infected with the human immunodeficiency virus. Anemia was present in 18% of asymptomatic patients who tested positive for the human immunodeficiency virus, 50% of patients with a condition related to the acquired immunodeficiency syndrome (AIDS), and 75% of patients with AIDS. The mean SIE level for untreated AIDS patients was greater than for patients who tested positive for human immunodeficiency virus or patients with an AIDS-related condition but not outside the normal range for SIE, and the incremental increase in SIE level for a given decline in hemoglobin level was much less in AIDS patients than in patients with uncomplicated iron deficiency anemia. Forty-two patients were treated with zidovudine, and the hemoglobin level fell 10 g/L or more in 48%. The data indicate that SIE level is inappropriately low in anemic AIDS patients. The ability of these patients to produce erythropoietin is intact and can be expressed with zidovudine therapy. However, even very high levels of SIE fail to stimulate erythropoiesis adequately.

Spivak, J.L.; Barnes, D.C.; Fuchs, E.; Quinn, T.C. (Johns Hopkins Univ. School of Medicine, Baltimore, MD (USA))

1989-06-02

218

Islet-1 Immunoreactivity in the Developing Retina of Xenopus laevis  

Science.gov (United States)

The LIM-homeodomain transcription factor Islet1 (Isl1) has been widely used as a marker of neuronal differentiation in the developing visual system of different classes of vertebrates, including mammals, birds, reptiles, and fish. In the present study, we analyzed the spatial and temporal distribution of Isl1-immunoreactive cells during Xenopus laevis retinal development and its relation to the formation of the retinal layers, and in combination with different markers of cell differentiation. The earliest Isl1 expression appeared at St29-30 in the cell nuclei of sparse differentiating neuroblasts located in the vitreal surface of the undifferentiated retina. At St35-36, abundant Isl1-positive cells accumulated at the vitreal surface of the neuroepithelium. As development proceeded and through the postmetamorphic juveniles, Isl1 expression was identified in subpopulations of ganglion cells and in subsets of amacrine, bipolar, and horizontal cells. These data together suggest a possible role for Isl1 in the early differentiation and maintenance of different retinal cell types, and Isl1 can serve as a specific molecular marker for the study of retinal cell specification in X. laevis.

Alvarez-Hernan, Guadalupe; Bejarano-Escobar, Ruth; Morona, Ruth; Gonzalez, Agustin; Martin-Partido, Gervasio

2013-01-01

219

Neurotensin-like immunoreactivity in the nervous system of hydra  

DEFF Research Database (Denmark)

Neurotensin-like immunoreactivity is found in nerve fibers present in all body regions of hydra. The nerve fibers are especially numerous in the ectoderm at the bases of the tentacles and in the ectoderm at a site just above the foot. Radioimmunoassays of acetic-acid extracts of hydra, using various region-specific antisera towards mammalian neurotensin, show the presence of multiple neurotensin-related peptides. The amounts of these peptides vary between 1 and 350 pmol per gram wet weight. Gel filtration on Sephadex G-25 reveals a fraction of neurotensin-like peptides that crossreact equally well with an antiserum directed against sequence 1-8 and an antiserum directed against sequence 6-13 of neurotensin. This fraction elutes also at the position of neurotensin and might closely resemble the mammalian peptide. A fraction eluting with the void volume crossreacts preferentially with antisera directed against sequences 1-8 and 10-13 of neurotensin. Several components of apparent lower molecular weight than neurotensin crossreact preferentially with an antiserum against sequence 10-13. These last peptides represent the major portion of the neurotensin-like peptides in hydra.

Grimmelikhuijzen, C J; Carraway, R E

1981-01-01

220

Inducible nitric oxide synthase immunoreactivity in healthy rat pancreas.  

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Full Text Available Nitric oxide (NO is produced by NO synthase (NOS isoforms: neuronal NOS (nNOS, endothelial NOS (eNOS and inducible NOS (iNOS. It is believed that, while nNOS and eNOS are effective in regulation of normal physiological processes, iNOS is expressed at an increasing rate especially in inflammatory process. The aim of this study was to determine the presence of iNOS immunoreactivity (iNOS-IR and, to compare the iNOS-IR in islet of Langerhans cells (LC, acinar cells (AC, centroacinar cells (CC and ductal cells (DC by immunohistochemical (IHC method in healthy rat pancreata. This study revealed the presence of iNOS-IR in all cell types except AC. Statistical analysis revealed a highly significant difference (p<0.001 with respect to iNOS-IR in comparison of all cell types. However, binary comparison of cell types revealed no significant differences between LC and DC (p=0.136, significant differences LC and CC, CC and DC (p=0.001 and 0.022, respectively and a highly significant differences LC and AC, AC and DC (P<0.001. The results of this study indicate that iNOS-IR is present in almost all LC. Thus, especially in reseach related to diabetes, it should not be disregarded that iNOS may be constitutively present in pancreatic islets.

Nurullah Keklikoglu

2008-06-01

 
 
 
 
221

Vasopressin-immunoreactive cell bodies in the bed nucleus of the stria terminalis of the rat.  

Science.gov (United States)

In the dorsal and ventral portions of the bed nucleus of the stria terminalis of the rat numerous cell bodies immunoreactive for vasopressin and neurophysin II were found after colchicin pretreatment. These cells are predominantly multipolar but sometimes also bipolar, and have a width and length of approximately 9 and 16 microns, respectively. In the homozygous Brattleboro rat, which is deficient in vasopressin, no immunoreactive vasopressin was found in these cells. Following incubation with anti-oxytocin and anti-bovine neurophysin I, only magnocellular immunoreactive cell bodies were found in the septal region. The consequences of these results concerning the vasopressin fiber pathways in the brain are discussed. PMID:6339062

van Leeuwen, F; Caffé, R

1983-01-01

222

FMRF-amide-like immunoreactivity in brain and pituitary of the hagfish Eptatretus burgeri (Cyclostomata)  

DEFF Research Database (Denmark)

Paraffin sections of brain and pituitary of the hagfish Eptatretus burgeri were immunostained with an antiserum to FMRF-amide. Immunoreactivity was visible in a large number of neurons in the posterior part of the ventromedial hypothalamus and in long neuronal processes extending cranially from the hypothalamus to the olfactory system and caudally to the medulla oblongata. FMRF-amide-like immunoreactivity was also found in cells of the adenohypophysis. These observations suggest that the hagfish possesses a brain FMRF-amide-like transmitter system and pituitary cells containing FMRF-amide-like material. Antisera to ACTH, alpha-MSH and pancreatic polypeptide gave no immunoreaction in hagfish brain or pituitary.

Jirikowski, G; Erhart, G

1984-01-01

223

Biochemical analysis of CTLA-4 immunoreactive material from human blood  

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Full Text Available Abstract Background CTLA-4 was initially described as a membrane-bound molecule that inhibited lymphocyte activation by interacting with B7.1 and B7.2 molecules on antigen presenting cells. Alternative splicing of mRNA encoding the CTLA-4 receptor leads to the production of a molecule (sCTLA-4 that lacks a membrane anchor and is therefore secreted into the extracellular space. Despite studies finding that people with autoimmune disease more frequently express high levels of sCTLA-4 in their blood than apparently healthy people, the significance of these findings is unclear. Methods Molecules isolated from blood using CTLA-4 specific antibodies were analyzed with ligand binding assays, mass spectroscopy, and biochemical fractionation in an effort to increase our understanding of CTLA-4 immunoreactive material. Results Mass spectroscopy analysis of the molecules recognized by multiple CTLA-4-specific antibodies failed to identify any CTLA-4 protein. Even though these molecules bind to the CTLA-4 receptors B7.1 and B7.2, they also exhibit properties common to immunoglobulins. Conclusion We have identified molecules in blood that are recognized by CTLA-4 specific antibodies but also exhibit properties of immunoglobulins. Our data indicates that what has been called sCTLA-4 is not a direct product of the CTLA-4 gene, and that the CTLA-4 protein is not part of this molecule. These results may explain why the relationship of sCTLA-4 to immune system activity has been difficult to elucidate.

Dennert Kate

2009-09-01

224

Clinicopathology significance of podoplanin immunoreactivity in esophageal squamous cell carcinoma  

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Backgroud and aim: Podoplanin (D2-40) is a specific marker for lymphatic endothelium. The vast majority of previous studies on podoplanin immunostaining in esophageal squamous cell carcinoma (ESCC) focused on identifying lymphatic vessel invasion (LVI) and counting lymphatic vessel density (LVD) and had contradictory results. Recent studies show podoplanin expression on cancer cells or tumor stroma in several cancers, which have specific significance; but the status in ESCC remains unclear. Therefore, the aim of this study was to further study and summarize the clinicopathological significance of podoplanin immunoreactivity in ESCC. Materials and methods: We examined podoplanin expression in tissue specimens from 107 patients with ESCC by immunohistochemistry. Podoplanin positive lymphatic vessels in intratumoral and peritumoral tissues and podoplanin positive expression in cancer cells and tumor stroma were analyzed, and correlated with clinicopathologic parameters and three-year overall and free-disease survival. Results: 34 (31.8%) and 28 (26.2%) of 107 specimens had podoplanin positive expression in cancer cells and tumor stroma, respectively. Logistic regression analysis showed high intratumoral lymphatic vessel density (I-LVD) and podoplanin positivity in cancer cells were increased risks of lymph node metastasis (LNM) (OR = 2.45, P = 0.03; OR = 0.35, P = 0.01, respectively). Survival analysis showed that I-LVD was a significant factor related to poor three-year overall and free-disease survival (P = 0.04, P = 0.03, respectively). Conclusions: Previous data and our results show that podoplanin seems to be a useful marker to predict LNM, recurrence, and worse prognosis in ESCC; in particular, LVI, high I-LVD, and podoplanin positivity in cancer cells are associated with LNM, recurrence and overall survival.

Ma, Wei; Wang, Kai; Yang, Shaoqi; Wang, Jianbo; Tan, Bingxu; Bai, Bing; Wang, Nana; Jia, Yibin; Jia, Ming; Cheng, Yufeng

2014-01-01

225

Comparison of immunoreactive serum trypsinogen and lipase in Cystic Fibrosis  

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The incidence of Cystic Fibrosis (CF) is 1 in 2,000. Early detection and treatment of CF may necessitate newborn screening with a reliable and cost-effective test. Serum immunoreactive trypsinogen (IRT) an enzyme produced by the pancreas, is detectable by radioimmunoassay (RIA) techniques. Recently, it has been shown that IRT is elevated in CF infants for the first few months of life and levels become subnormal as pancreatic insufficiency progresses. Other enzymes produced by the pancreas, such as lipase, are also elevated during this time. The author's earlier work confirmed previous reports of elevated IRT levels in CF infants. The development of a new RIA for lipase (nuclipase) has enabled comparison of these 2 pancreatic enzymes in C.F. Serum IRT and lipase determinations were performed on 2 groups of CF patients; infants under 1 year of age, and children between 1 and 18 years of age. Control populations of the same age groups were included. The results showed that both trypsin (161 +- 92 ng/ml, range 20 to 400) and lipase (167 +- 151 ng/ml, range 29 to 500) are elevated in CF in the majority of infants. Control infants had values of IRT ranging from 20 to 29.5 ng/ml and lipase values ranging from 23 to 34 ng/ml. IRT becomes subnormal in most CF patients by 8 years of age as pancreatic function insufficiency increases. Lipase levels and IRT levels correlate well in infancy, but IRT is a more sensitive indicator of pancreatic insufficiency in older patients with CF

1984-06-05

226

Induction of Fos protein immunoreactivity by spinal cord contusion  

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Full Text Available The objective of the present study was to identify neurons in the central nervous system that respond to spinal contusion injury in the rat by monitoring the expression of the nuclear protein encoded by the c-fos gene, an activity-dependent gene, in spinal cord and brainstem regions. Rats were anesthetized with urethane and the injury was produced by dropping a 5-g weight from 20.0 cm onto the exposed dura at the T10-L1 vertebral level (contusion group. The spinal cord was exposed but not lesioned in anesthetized control animals (laminectomy group; intact animals were also subjected to anesthesia (intact control. Behavioral alterations were analyzed by Tarlov/Bohlman scores, 2 h after the procedures and the animals were then perfused for immunocytochemistry. The patterns of Fos-like immunoreactivity (FLI which were site-specific, reproducible and correlated with spinal laminae that respond predominantly to noxious stimulation or injury: laminae I-II (outer substantia gelatinosa and X and the nucleus of the intermediolateral cell column. At the brain stem level FLI was detected in the reticular formation, area postrema and solitary tract nucleus of lesioned animals. No Fos staining was detected by immunocytochemistry in the intact control group. However, detection of FLI in the group submitted to anesthesia and surgical procedures, although less intense than in the lesion group, indicated that microtraumas may occur which are not detected by the Tarlov/Bohlman scores. There is both a local and remote effect of a distal contusion on the spinal cord of rats, implicating sensory neurons and centers related to autonomic control in the reaction to this kind of injury.

Del-Bel E.A.

2000-01-01

227

Induction of Fos protein immunoreactivity by spinal cord contusion  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english The objective of the present study was to identify neurons in the central nervous system that respond to spinal contusion injury in the rat by monitoring the expression of the nuclear protein encoded by the c-fos gene, an activity-dependent gene, in spinal cord and brainstem regions. Rats were anest [...] hetized with urethane and the injury was produced by dropping a 5-g weight from 20.0 cm onto the exposed dura at the T10-L1 vertebral level (contusion group). The spinal cord was exposed but not lesioned in anesthetized control animals (laminectomy group); intact animals were also subjected to anesthesia (intact control). Behavioral alterations were analyzed by Tarlov/Bohlman scores, 2 h after the procedures and the animals were then perfused for immunocytochemistry. The patterns of Fos-like immunoreactivity (FLI) which were site-specific, reproducible and correlated with spinal laminae that respond predominantly to noxious stimulation or injury: laminae I-II (outer substantia gelatinosa) and X and the nucleus of the intermediolateral cell column. At the brain stem level FLI was detected in the reticular formation, area postrema and solitary tract nucleus of lesioned animals. No Fos staining was detected by immunocytochemistry in the intact control group. However, detection of FLI in the group submitted to anesthesia and surgical procedures, although less intense than in the lesion group, indicated that microtraumas may occur which are not detected by the Tarlov/Bohlman scores. There is both a local and remote effect of a distal contusion on the spinal cord of rats, implicating sensory neurons and centers related to autonomic control in the reaction to this kind of injury.

E.A., Del-Bel; C.A.G., Borges; H.L.A., Defino; F.S., Guimarães.

228

Diurnal variation of ?-endorphin like immunoreactivity in rat brain, pituitary gland, and plasma  

International Nuclear Information System (INIS)

?-endorphin like immunoreactivity was measured in the brain, pituitary gland and plasma of rats at 2 A.M, 8 A.M, 2 P.M and 8 P.M. Values were higher in the brain and pituitary gland at 8 P.M and in the plasma at 8 A.M and 2 P.M. The findings suggest a circadian rhythm in the production and release of ?-endorphin immunoreactive material. (Author)

1984-01-01

229

Compartmental distribution of striatal cell bodies expressing [Met]enkephalin-like immunoreactivity.  

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Striatal cell bodies and fibers expressing [Met]enkephalin [( Met]Enk)-like immunoreactivity were studied with two variants of the peroxidase-antiperoxidase method in normal primates and cats and in cats pretreated with colchicine. Strikingly different patterns of [Met]Enk-like immunoreactivity were observed, both in fiber and cell body immunostaining, depending on the technical protocols followed; no single histochemical protocol fully revealed the compartmentalization present. In the dorsal...

Graybiel, A. M.; Chesselet, M. F.

1984-01-01

230

Proenkephalin, [Met]enkephalin, and oxytocin immunoreactivities are colocalized in bovine hypothalamic magnocellular neurons.  

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The distribution of proenkephalin and [Met]enkephalin immunoreactivities in the bovine hypothalamo-neurohypophyseal system was studied by use of specific antisera. Proenkephalin and [Met]enkephalin immunoreactivities were found in magnocellular neuronal cell bodies in the dorsal part of the supraoptic nuclei and in the peripheral part of the paraventricular nuclei. A densely staining network of nerve terminals was found in the external part of the median eminence and in the posterior hypophys...

Vanderhaeghen, J. J.; Lotstra, F.; Liston, D. R.; Rossier, J.

1983-01-01

231

Immunocytochemical localization of enkephalin-like immunoreactivity in the retina of the guinea pig.  

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The distribution of enkephalin-like immunoreactivity in the retina of the guinea pig was studied. Indirect immunofluorescence techniques were used on retinae with and without colchicine pretreatment. In retinae not receiving colchicine pretreatment, enkephalin-like immunoreactivity was seen in fibers in the inner plexiform layer, predominantly in laminae 1, 3, and 5. In colchicine-pretreated retinae, enkephalin immunofluorescent cell bodies were seen in the inner margin of the inner nuclear l...

Altschuler, R. A.; Mosinger, J. L.; Hoffman, D. W.; Parakkal, M. H.

1982-01-01

232

Prognostic significance of CEA immunoreactivity patterns in large bowel carcinoma tissue.  

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In order to determine the clinical value of CEA detection in large bowel cancer tissue the patterns rather than the intensity of immunoreactivity of CEA reactive antibodies were analyzed in 312 large bowel cancer patients especially in relation to patient survival. CEA immunoreactivity appeared to be distinguishable into a predominantly apical/cytoplasmic and a predominantly membranous pattern. Twenty-four (7.7%) tumours were found to be CEA negative or only focally positive. Two hundred and ...

Wiggers, T.; Arends, J. W.; Verstijnen, C.; Moerkerk, P. M.; Bosman, F. T.

1986-01-01

233

Distribution of NGF and NT-3-like protein immunoreactivity in the teleost kidney.  

Science.gov (United States)

By means of immunochemistry and immunohistochemistry, we investigated in the kidney of freshwater and marine teleostean species for the presence and localization of three neurotrophins: nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin (NT)-3. In both species studied, NGF-like and NT-3-like immunoreactivity were present in the kidney with different distribution patterns, while BDNF-like immunoreactivity was never detected. In goldfish, NGF-like and NT-3-like immunoreactivity were identified extensively in cells along part of the arterial branches adjacent to the afferent arterioles. In scorpion fish, NGF-like and NT-3-like immunoreactive cells were observed both on afferent arterioles and on adjacent secondary branches derived from renal arteries. No immunoreactivity was detected in other renal structures. A staining pattern of immunoreactivity similar to that obtained for NGF and NT-3 was detected utilizing S100 antibody as a juxtaglomerular (JG) cell marker. Double immunolabellings NGF/S100 and NT-3/S100 evidenced the coexistence of neurotrophin-like proteins and S100-like protein in the same immunoreactive cells, thus identifying them as juxtaglomerular cells. Western blot analysis revealed the presence of molecules immunoreactive to NGF and NT-3, whose molecular weights were very similar to those of the corresponding mammalian neurotrophins. These findings extend the presence and distribution of NGF-like and NT-3-like IR in the kidney to teleost species, suggesting a probable participation of these proteins in the renal functions of freshwater and marine teleosts. PMID:15816034

Arcamone, Nadia; Lucini, Carla; Borzacchiello, Giuseppe; Castaldo, Luciana; Gargiulo, Giuliana; De Girolamo, Paolo

2005-01-01

234

Decreased serotonin transporter immunoreactivity in the human hypothalamic infundibular nucleus of overweight subjects  

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Full Text Available Context: That serotonin plays a role in the regulation of feeding behavior and energy metabolism has been known for a long time. Serotonin transporters (SERT play a crucial role in serotonin signalling by regulating its availability in the synaptic cleft. The neuroanatomy underlying serotonergic signalling in humans is largely unknown, and until now, SERT immunoreactivity in relation to body weight has not been investigated.Objective: To clarify the distribution of SERT immunoreactivity throughout the human hypothalamus and to compare SERT immunoreactivity in the infundibular nucleus (IFN, the human equivalent of the arcuate nucleus, in lean and overweight subjects. Design: First, we investigated the distribution of serotonin transporters (SERT over the rostro-caudal axis of six postmortem hypothalami by means of immunohistochemistry. Second, we estimated SERT immunoreactivity in the IFN of lean and overweight subjects. Lastly, double-labelling of SERT with Neuropeptide Y (NPY and melanocortin cell populations was performed to further identify cells showing basket-like SERT staining. Results: SERT-immunoreactivity was ubiquitously expressed in fibers throughout the hypothalamus and was the strongest in the IFN. Immunoreactivity in the IFN was lower in overweight subjects (p=0.036. Basket-like staining in the IFN was highly suggestive of synaptic innervation. A very small minority of cells showed SERT double labelling with NPY, agouti-related protein and ??melanocyte stimulating hormone. Conclusions: SERT is ubiquitously expressed in the human hypothalamus. Strong SERT immunoreactivity, was observed in the IFN a region important for appetite regulation, in combination with lower SERT immunoreactivity in the IFN of overweight and obese subjects, may point towards a role for hypothalamic SERT in human obesity.

AnnekeAlkemade

2014-05-01

235

Levels of immunoreactive inhibin-like material in urine during the menstrual cycle  

International Nuclear Information System (INIS)

Using a specific and sensitive radioimmunoassay, the authors determined levels of inhibinlike material in the urine of eight healthy women with normal menstrual cycle length of 28 +- 4 days. The results revealed a cyclic variation in urinary immunoreactive inhibin levels during the menstrual cycles, with a sharp rise in levels three to four days prior to luteinizing hormone (LH) and follicle-stimulating hormone (FSH) peaks. These levels of immunoreactive inhibin may thus serve as a parameter to detect impending LH surge. (author)

1983-12-01

236

The development of somatostatin immunoreactivity in the interpeduncular nucleus of the cat.  

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Somatostatin (SS) immunoreactivity was localized in a population of neurons in the interpeduncular nucleus (IPN) of the developing and adult cat. SS-immunoreactive perikarya and fibers were found in several subdivisions of the IPN, but were most heavily concentrated in the central nucleus (IPC). Some immunoreactive fibers appeared to be associated with the ventral surface of the IPN where they may come in contact with the interpeduncular cistern. Bundles of immunoreactive fibers also appeared to leave the rostral pole of the IPN bilaterally. Still other SS-positive perikarya and fibers were associated with the tegmentopeduncular tract and the medial division of the dorsal tegmental nucleus. SS was present in both perikarya and fibers of the IPN at birth. The concentration of immunoreactivity increased dramatically during the first 100 days postnatal, but by 180 days the overall intensity of immunoreactivity had decreased to adult levels. These data indicate that SS may be present in the cat IPN in a specific population of neurons that is most active during early postnatal development. The data are consistent with previous suggestions that SS plays a special role in the development of certain neuronal systems and may be particularly important in the integration of behaviors during the neonatal period. PMID:2862961

Morley, B J; Spangler, K M; Javel, E

1985-06-01

237

Cardiac abnormalities in liver cirrhosis.  

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Cirrhosis is associated with several circulatory abnormalities. A hyperkinetic circulation characterized by increased cardiac output and decreased arterial pressure and peripheral resistance is typical. Despite this hyperkinetic circulation, some patients with alcoholic cirrhosis have subclinical cardiomyopathy with evidence of abnormal ventricular function unmasked by physiologic or pharmacologic stress. Florid congestive alcoholic cardiomyopathy develops in a small percentage, but the concu...

Lee, S. S.

1989-01-01

238

Orthopaedic abnormalities in primary myopathies.  

Science.gov (United States)

Orthopaedic abnormalities are frequently recognised in patients with myopathy but are hardly systematically reviewed with regard to type of myopathy, type of orthopaedic problem, and orthopaedic management. This review aims to summarize recent findings and current knowledge about orthopaedic abnormalities in these patients, their frequency, and possible therapeutic interventions. A MEDLINE search for the combination of specific terms was carried out and appropriate articles were reviewed for the type of myopathy, types of orthopaedic abnormalities, frequency of orthopaedic abnormalities, and possible therapeutic interventions. Orthopaedic abnormalities in myopathies can be most simply classified according to the anatomical location into those of: the spine, including dropped head, camptocormia, scoliosis, hyperlordosis, hyperkyphosis, or rigid spine; the thorax, including pectus excavatum (cobbler's chest), anterior/posterior flattening, or pectus carinatum (pigeon's chest); the limb girdles, including scapular winging and pelvic deformities; and the extremities, including contractures, hyperlaxity of joints, and hand or foot deformities. These orthopaedic abnormalities can be most frequently found in arthrogryposis, muscular dystrophies, congenital myopathies, myofibrillar myopathies, and myotonic dystrophies. Occasionally, they also occur in metabolic myopathies or other types of myopathy. Most of the orthopaedic abnormalities are sufficiently accessible to conservative or surgical orthopaedic treatment. Orthopaedic abnormalities have major implications in the management and outcome of myopathy patients; they should be closely monitored and treated on time. PMID:22187829

Finsterer, Josef; Strobl, Walter

2011-10-01

239

Slc4a11 gene disruption in mice: cellular targets of sensorineuronal abnormalities.  

Science.gov (United States)

NaBC1 (the SLC4A11 gene) belongs to the SLC4 family of sodium-coupled bicarbonate (carbonate) transporter proteins and functions as an electrogenic sodium borate cotransporter. Mutations in SLC4A11 cause either corneal abnormalities (corneal hereditary dystrophy type 2) or a combined auditory and visual impairment (Harboyan syndrome). The role of NaBC1 in sensory systems is poorly understood, given the difficulty of studying patients with NaBC1 mutations. We report our findings in Slc4a11(-/-) mice generated to investigate the role of NaBC1 in sensorineural systems. In wild-type mice, specific NaBC1 immunoreactivity was detected in fibrocytes of the spiral ligament, from the basal to the apical portion of the cochlea. NaBC1 immunoreactivity was present in the vestibular labyrinth, in stromal cells underneath the non-immunoreactive sensory epithelia of the macula utricle, sacule, and crista ampullaris, and the membranous vestibular labyrinth was collapsed. Both auditory brain response and vestibular evoked potential waveforms were significantly abnormal in Slc4a11(-/-) mice. In the cornea, NaBC1 was highly expressed in the endothelial cell layer with less staining in epithelial cells. However, unlike humans, the corneal phenotype was mild with a normal slit lamp evaluation. Corneal endothelial cells were morphologically normal; however, both the absolute height of the corneal basal epithelial cells and the relative basal epithelial cell/total corneal thickness were significantly increased in Slc4a11(-/-) mice. Our results demonstrate for the first time the importance of NaBC1 in the audio-vestibular system and provide support for the hypothesis that SLC4A11 should be considered a potential candidate gene in patients with isolated sensorineural vestibular hearing abnormalities. PMID:19586905

Lopez, Ivan A; Rosenblatt, Mark I; Kim, Charles; Galbraith, Gary C; Jones, Sherri M; Kao, Liyo; Newman, Debra; Liu, Weixin; Yeh, Stacey; Pushkin, Alexander; Abuladze, Natalia; Kurtz, Ira

2009-09-25

240

Chromosomal abnormalities in human sperm  

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The ability to analyze human sperm chromosome complements after penetration of zona pellucida-free hamster eggs provides the first opportunity to study the frequency and type of chromosomal abnormalities in human gametes. Two large-scale studies have provided information on normal men. We have studied 1,426 sperm complements from 45 normal men and found an abnormality rate of 8.9%. Brandriff et al. (5) found 8.1% abnormal complements in 909 sperm from 4 men. The distribution of numerical and structural abnormalities was markedly dissimilar in the 2 studies. The frequency of aneuploidy was 5% in our sample and only 1.6% in Brandriff's, perhaps reflecting individual variability among donors. The frequency of 24,YY sperm was low: 0/1,426 and 1/909. This suggests that the estimates of nondisjunction based on fluorescent Y body data (1% to 5%) are not accurate. We have also studied men at increased risk of sperm chromosomal abnormalities. The frequency of chromosomally unbalanced sperm in 6 men heterozygous for structural abnormalities varied dramatically: 77% for t11;22, 32% for t6;14, 19% for t5;18, 13% for t14;21, and 0% for inv 3 and 7. We have also studied 13 cancer patients before and after radiotherapy and demonstrated a significant dose-dependent increase of sperm chromosome abnormalities (numerical and structural) 36 months after radiation treatment.

Martin, R.H.

1985-01-01

 
 
 
 
241

Plasma immunoreactive neuropeptide Y in congestive heart failure at rest and during exercise.  

DEFF Research Database (Denmark)

The purpose of the study described here was to study plasma immunoreactive Neuropeptide Y (NPY) at rest and during exercise in patients with congestive heart failure (CHF) and in healthy subjects. Thirty-five patients, mean age 64 years, with CHF in optimal treatment and with a mean ejection fraction of 32%, were studied at rest and during exercise. Twelve age and sex matched healthy subjects were compared for resting values. Another nine healthy subjects were studied at rest and during exercise at a constant low load of 75W and at a high load defined as 80% of their individual maximal capacity. In patients with congestive heart failure mean plasma immunoreactive NPY at rest was 10.3 pmol l-1 and was not significantly different from the control group. No differences between patients with slight and severe CHF were found and there was no correlation between plasma immunoreactive NPY and left ventricular ejection fraction. Mean maximal exercise time was on average 6.3 min. Only three patients exercised more than 10 min. At maximal exercise mean plasma immunoreactive NPY was 10.6 pmol l-1 the same as at rest. Plasma noradrenaline was increased in CHF patients compared to healthy subjects, and rose further during exercise. In healthy subjects plasma immunoreactive NPY rose significantly on both workloads, but more on the high load (p < 0.05), when the rise was first significant after 10 min. Plasma immunoreactive NPY at rest and during exercise was not increased in CHF patients in optimal medical treatment. Consequently plasma immunoreactive NPY is not a useful marker of the severity of CHF in these patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Madsen, B K; Husum, D

1993-01-01

242

Pregnancy Complications: Umbilical Cord Abnormalities  

Science.gov (United States)

... defects . These tests may include a detailed ultrasound, amniocentesis (to check for chromosomal abnormalities) and in some ... the provider may recommend additional tests, such as amniocentesis and a detailed ultrasound, to diagnose or rule ...

243

The abnormal proximal tibiofibular joint.  

Science.gov (United States)

Abnormalities of the proximal tibiofibular joint are infrequently encountered. Mostly instability occurs as a result of trauma. Four types of instability are distinguished: subluxation, anterolateral, posteromedial and superior dislocation. Four radiological methods designed to visualize abnormalities of the proximal tibiofibular joint are discussed. Special notice is given to the clinical relevance of a new method. Instability was demonstrated in 19 patients; 16 of these were treated by an operation. History, data obtained by physical examination and ways of treatment are discussed. PMID:6703874

Veth, R P; Kingma, L M; Nielsen, H K

1984-01-01

244

Calbindin, calretinin and parvalbumin immunoreactivity in the retina of the chameleon (Chamaeleo chamaeleon).  

Science.gov (United States)

Apart from the pioneering studies of Ramon y Cajal [1893] and Rochon-Duvigneaud [1943], few studies have been devoted to the detailed study of the cytological and biochemical structure of the chameleon retina. In the present study we analyzed the expression of calbindin (CB), calretinin (CR) and parvalbumin (PV) immunoreactivities in the chameleon retina, and compared their distribution with those found in the retinas of other vertebrate species. CB immunoreactivity is dense in photoreceptors, horizontal and some lower amacrine cells. The most intense immunoreactivity was observed for calretinin; CR-ir amacrine cells are distributed throughout the inner nuclear, inner plexiform, and ganglion cell layers of the retina. Horizontal cells also display immunoreactivity to CR. A few retinal interneurons are weakly PV-ir. Double-labeling shows that all PV-ir or CB-ir cells, except the photoreceptors, are also strongly CR-ir. The distributions of these calcium-binding proteins in the chameleon retina share similarities with those observed in mammalian and avian retinas. In addition, the widespread distribution and co-localization of CB and CR reinforces the idea that these proteins play a general role in buffering the intracellular calcium levels in retinal cells. Furthermore, CB- and CR-immunoreactivities have enabled us to identify for the first time axon-bearing horizontal cells in the peripheral retina of the chameleon, very similar to those described in mammals. PMID:15687725

Bennis, M; Versaux-Botteri, C; Repérant, J; Armengol, J A

2005-01-01

245

Compartmental distribution of striatal cell bodies expressing [Met]enkephalin-like immunoreactivity.  

Science.gov (United States)

Striatal cell bodies and fibers expressing [Met]enkephalin [( Met]Enk)-like immunoreactivity were studied with two variants of the peroxidase-antiperoxidase method in normal primates and cats and in cats pretreated with colchicine. Strikingly different patterns of [Met]Enk-like immunoreactivity were observed, both in fiber and cell body immunostaining, depending on the technical protocols followed; no single histochemical protocol fully revealed the compartmentalization present. In the dorsal striatum, patches of [Met]Enk-positive neuropil, known to line up with the acetylcholinesterase-poor striatal zones called striosomes, appeared in sections treated by protocols favoring fiber immunostaining. In sections stained by procedures favoring perikaryal staining, the striosomes appeared as Enk-poor patches in a field of immunoreactive cells and neuropil. When cell-body staining was enhanced by pretreatment with colchicine, cells expressing [Met]Enk-like immunoreactivity appeared both in and out of striosomes, and the striosomal neuropil appeared Enk-rich. These results suggest that there are subtypes of Enk-positive neurons in the striatum, including a "colchicine-dependent subtype" in dorsal striosomes, and suggest that the Enk-positive striatal neuropil is also made up of different components. Immunospecificity of this dorsal striosomal system was further demonstrated by the finding that neurons expressing intense immunoreactivity to substance P and to dynorphin B were largely confined to striosomes. Images

Graybiel, A M; Chesselet, M F

1984-01-01

246

Localisation of atrial natriuretic peptide immunoreactivity in the ventricular myocardium and conduction system of the human fetal and adult heart.  

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Atrial natriuretic peptide immunoreactivity was found in ventricular and atrial tissues with specific antisera raised to the amino and carboxy terminal regions of the precursor molecule. In 13 developing human hearts (7-24 weeks' gestation) the immunoreactivity was concentrated in the atrial myocardium and ventricular conduction system but it was also detected in the early fetal ventricular myocardium. Immunoreactivity in five normal adults was largely confined to the atrial myocardium althou...

Wharton, J.; Anderson, R. H.; Springall, D.; Power, R. F.; Rose, M.; Smith, A.; Espejo, R.; Khaghani, A.; Wallwork, J.; Yacoub, M. H.

1988-01-01

247

Comparative tumour localization properties of radiolabelled monoclonal antibody preparations of defined immunoreactivities  

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The immunoreactive fraction of an anti-CEA monoclonal antibody preparation has been progressively decreased by the addition of increasing proportions of impurity in the form of immunologically inert mouse immunoglobulin. Following radioiodination, the immunoreactive fractions of the preparations were determined and their localization in a human tumour xenograft in nude mice was assessed. There was a progressive decline in tumour localization, from tumour to blood ratios of 2:1 with unadulterated antibody to 0.6:1 with preparations only 15% with respect to the initial antibody. These findings demonstrate that the immunoreactive fraction of monoclonal antibody preparations is a major limiting factor in tumour localization and this has implications for experimental and clinical applications of monoclonal antibodies.

Pimm, M.V.; Baldwin, R.W.

1987-10-01

248

Comparative tumour localization properties of radiolabelled monoclonal antibody preparations of defined immunoreactivities  

International Nuclear Information System (INIS)

The immunoreactive fraction of an anti-CEA monoclonal antibody preparation has been progressively decreased by the addition of increasing proportions of impurity in the form of immunologically inert mouse immunoglobulin. Following radioiodination, the immunoreactive fractions of the preparations were determined and their localization in a human tumour xenograft in nude mice was assessed. There was a progressive decline in tumour localization, from tumour to blood ratios of 2:1 with unadulterated antibody to 0.6:1 with preparations only 15% with respect to the initial antibody. These findings demonstrate that the immunoreactive fraction of monoclonal antibody preparations is a major limiting factor in tumour localization and this has implications for experimental and clinical applications of monoclonal antibodies. (orig.)

1987-01-01

249

Morphometric characteristics of Neuropeptide Y immunoreactive neurons of human cortical amygdaloid nucleus  

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Full Text Available Introduction Cortical amygdaloid nucleus belongs to the corticomedial part of the amygdaloid complex. In this nucleus there are neurons that produce neuropetide Y. This peptide has important roles in sleeping, learning, memory, gastrointestinal regulation, anxiety, epilepsy, alcoholism and depression. Material and methods We investigated morphometric characteristics (numbers of primary dendrites, longer and shorter diameters of cell bodies and maximal radius of dendritic arborization of NPY immunoreactive neurons of human cortical amygdaloid nucleus on 6 male adult human brains, aged 46 to 77 years, by immunohistochemical avidin-biotin technique. Results Our investigation has shown that in this nucleus there is a moderate number of NPY immunoreactive neurons. 67% of found neurons were nonpyramidal, while 33% were pyramidal. Among the nonpyramidal neurons the dominant groups were multipolar neurons (41% - of which 25% were multipolar irregular, and 16% multipolar oval. Among the pyramidal neurons the dominant groups were the neurons with triangular shape of cell body (21%. All found NPY immunoreactive neurons (pyramidal and nonpyramidal altogether had intervals of values of numbers of primary dendrites 2 to 6, longer diameters of cell bodies 13 to 38 µm, shorter diameters of cell bodies 9 to 20 µm and maximal radius of dendritic arborization 50 to 340 µm. More than a half of investigated neurons (57% had 3 primary dendrites. Discussion and conclusion The other researchers did not find such percentage of pyramidal immunoreactive neurons in this amygdaloid nucleus. If we compare our results with the results of the ather researchers we can conclude that all pyramidal NPY immunoreactive neurons found in this human amygdaloid nucleus belong to the class I of neurons, and that all nonpyramidal NPY immunoreactive neurons belong to the class II of neurons described by other researchers. We suppose that all found pyramidal neurons were projectional.

Mališ Miloš

2008-01-01

250

SPERM ABNORMALITIES AND ITS TREATMENT  

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Full Text Available The term sperm is derived from the Greek word sperma means "seed" and it refers to the male reproductive cells. In the types of sexual reproduction known as anisogamy and oogamy, there is a marked difference in the size of the gametes with the smaller one being termed the "male" or sperm cell. A uniflagellar sperm cell that is motile is referred to as a spermatozoon, whereas a non-motile sperm cell is referred to as a spermatium. Sperm cells cannot divide and have a limited life span, but after fusion with egg cells during fertilization, a new organism begins developing, starting as a totipotent zygote. Sperm morphology the size and shape of sperm is checked as part of a standard semen analysis for male infertility. Many different types of sperm abnormalities occur. A common classification scheme is based on the location of the abnormalities. Those that are located in the sperm head are classified as primary. Abnormalities associated with neck, midpiece or tail are classified as secondary abnormalities. Included in the secondary abnormalities is the presence of cytoplasmic droplets.

Prajapati Parimal M.

2011-11-01

251

[Cytoskeletal proteins abnormalities in olivopontocerebellar atrophy].  

Science.gov (United States)

A highly sensitive silver technique for glial cytoplasmic inclusions in olivopontocerebellar atrophy (OPCA) was applied to 15 subjects with neurodegenerative disorders including 4 patients with OPCA, 4 patients with Joseph disease and with 3 normal control subjects, and the argyrophilic structures in the OPCA cases were immunocytochemically examined. As a result, the argyrophilic structures were found in the OPCA cases and Alzheimer cases. The argyrophilic structures included the white matter oligodendroglia, and neurons, astroglia and oligodendroglia in the putamen, pontine nucleus and inferior olivary nucleus. The pontocerebellar tracts and the surrounding white matter of the inferior olivary nucleus contained a small number of argyrophilic nerve fibers with a hollow structure, which were interpreted as myelin. Immunocytochemistry demonstrated that the oligodendroglia in the white matter had immunoreactivities to an paired helical filament (PHF), microtubule associated protein 5 (MAP5), tau and ubiquitin antiserum, and the astroglia in the white matters had an immunoreactivity to a MAP5 antiserum. In the putamen, pontine nucleus and inferior olivary nucleus, in addition to the immunoreactivities observed in the oligodendroglia, the neurons were immunoreactive for PHF, MAP5, tau and ubiquitin antisera, and the astroglia had the same immunoreactivities as the neurons except for being tau negative in the putamen. The nerve fibers in the pontocerebellar tract and inferior olivary nucleus capsule were strongly positive for myelin basic protein and negative for PHF antiserum. These findings indicate that the Gallyas positive argyrophilia in the OPCA subjects is closely associated with PHF or tau. PMID:1591102

Kobayashi, K; Miyazu, K; Katsukawa, K; Fukutani, Y; Mukai, M; Nakamura, I; Yamaguchi, N; Matsubara, R; Isaki, K

1992-03-01

252

Craniocervical junction abnormalities in dogs.  

Science.gov (United States)

Craniocervical junction abnormality (CJA) is a term that encompasses a number of developmental anatomical aberrations at the region of the caudal occiput and first two cervical vertebrae. Chiari-like malformation appears to be the most common CJA encountered in dogs, and there has been a tremendous amount of clinical investigation into this disorder in recent years. Other abnormalities in this region include atlanto-occipital overlap, dorsal constriction at C1/C2 and atlantoaxial instability. This review article presents an overview of the current understanding of CJA in dogs, as well as medical and surgical treatment options available. PMID:23556552

Dewey, C W; Marino, D J; Loughin, C A

2013-07-01

253

Seasonal changes in beta-endorphin-like immunoreactivity in the olfactory system of the female catfish, Clarias batrachus (Linn).  

Science.gov (United States)

In the olfactory system of the catfish Clarias batrachus, beta-endorphin-like immunoreactivity was seen in several olfactory receptor neurons (ORN) and their fiber projections extending caudally over the olfactory nerve to the olfactory bulb (OB). With beta-endorphin-like immunoreactivity as a cellular marker, the olfactory system in the female fish was investigated at different stages of its annual reproductive cycle. The reproductive cycle of the fish is divisible into four distinct phases: preparatory (February-April), prespawning (May-June), spawning (July-August), and postspawning (September-January). The gonosomatic index and the immunocytochemical profile of beta-endorphin-like immunoreactivity showed distinct changes as the fish progressed from one phase to another. In the preparatory phase, limited immunoreactivity was seen in the periphery of the bulb. However, the immunoreactivity showed a robust increase as the immunolabeled fibers extended progressively deeper into the bulb toward the mitral cell layer during the prespawning and spawning phases. Significant reduction in the immunoreactivity was noticed in the olfactory nerve layer of the fish in the postspawning phase. Several granule cells showed poor to moderate immunoreactivity during the spawning phase, although no immunoreactivity was seen in the inner cell layer during the rest of the year. The beta-endorphin-like immunoreactivity in the ORN also showed season-related changes, although these were less distinct. Whereas weak immunoreactivity confined to a few ORN was noticed in the fish collected in the preparatory phase, those in the prespawning phase showed conspicuous augmentation in immunoreactivity. During the spawning phase, the sensory layer of the olfactory epithelium showed reduced, homogenous immunoreactivity. In the postspawning phase, several ORN revealed distinct granular immunoreactivity, suggesting possibilities of de novo synthesis. These annual cyclic changes in the beta-endorphin-like immunoreactivity were consistently observed over a 30-month study period that spanned three consecutive spawning phases. The results suggest that the beta-endorphin-containing ORN, their fiber projections to the OB, and several granule cells in the inner cell layer may be involved in the processing of reproduction/reproductive behavior-related signals. PMID:11482933

Sarkar, S; Subhedar, N

2001-08-01

254

Interpreting chromosomal abnormalities using Prolog.  

Science.gov (United States)

This paper describes an expert system for interpreting the standard notation used to represent human chromosomal abnormalities, namely, the International System for Human Cytogenetic Nomenclature. Written in Prolog, this program is very powerful, easy to maintain, and portable. The system can be used as a front end to any database that employs cytogenetic notation, such as a patient registry. PMID:2185921

Cooper, G; Friedman, J M

1990-04-01

255

[Alterations of immunoreactivity in cnemical production workers in dependence on dose exposure to toxicants].  

Science.gov (United States)

The immunological examination of employees working in the production of caustic soda, vinyl chloride, polyvinyl chloride and epichlorohydrin has been performed. The features of the relationship between alterations in indices of immunoreactivity and dose exposure for total years of employment in persons exposed to various chemical substances have been established. PMID:23082668

2012-01-01

256

Immunoreactive anionic and cationic trypsins in serum after experimental porcine pancreatic transplantation.  

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Immunoreactive anionic and cationic trypsins (irAT and irCT) were measured in blood samples taken daily from 20 pigs during 60 days after whole organ pancreatic transplantation. Immunosuppression was discontinued on the twenty-eighth day after transplantation. IrAT concentrations showed three distinct peaks, the first immediately after operation, the second around the seventh day after operation, and the third irAT peak was seen 2 to 9 days after discontinuation of immunosuppression therapy. IrCT levels paralleled the irAT levels only in the first peak immediately after transplantation. After this there was a dissociation between the concentrations of the two immunoreactive trypsins, with low irCT levels and high irAT levels. Increased irAT levels heralded rejection of the pancreatic allograft by 4 to 30 days (median 20 days) in 13 of the 16 rejecting animals (80%). In addition, low irCT levels preceded hyperglycemia in 13 of the 16 rejecting pigs by 2 to 47 days (median 21 days). The first postoperative peak of immunoreactive trypsins is thought to be related to the operative and storage trauma, whereas the second and third peaks are thought to reflect a rejection process. The results suggest that immunoreactive trypsins can be used as markers for pancreatic allograft rejection. PMID:3535146

Borgström, A; Marks, W H; Dafoe, D C; Campbell, D A; Turcotte, J G

1986-11-01

257

Monoclonal antibody to the rat glucocorticoid receptor. Relationship between the immunoreactive and DNA-binding domain  

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The region of the glucocorticoid receptor that reacted with a monoclonal antibody (BUGR-1) was identified. In order to identify the immunoreactive region, the rat liver glucocorticoid receptor was subjected to limited proteolysis; immunoreactive fragments were identified by Western blotting. The monoclonal antibody reacted with both the undigested Mr approximately 97,000 receptor subunit and a Mr approximately 45,000 fragment containing the steroid-binding and DNA-binding domains. Digestion by trypsin also produced two steroid-binding fragments of Mr approximately 27,000 and 31,000 which did not react with the antibody and an immunoreactive Mr approximately 16,000 fragment. This Mr approximately 16,000 fragment was shown to bind to DNA-cellulose, indicating that it contained a DNA-binding domain of the receptor. The undigested receptor must have steroid associated with it to undergo activation to a DNA-binding form. However, the Mr approximately 16,000 immunoreactive fragment binds to DNA-cellulose even if it is obtained by digestion of the steroid-free holoreceptor which does not itself bind to DNA.

Eisen, L.P.; Reichman, M.E.; Thompson, E.B.; Gametchu, B.; Harrison, R.W.; Eisen, H.J.

1985-09-25

258

[The significance of digoxin-like immunoreactive factor (DLIF) for intensive care medicine].  

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The estimation of serum digoxin is a usual method in intensive care. In a case report the detection of digoxin-like-immunoreactive-factor (DLIF) is shown, which gives false high levels. DLIF is observed in renal damage, high cardiac activity, pregnancy and newborn. PMID:2168132

Simon, H B; Behrendt, W; Stein, T

1990-06-01

259

Endogenous digoxin-like immunoreactive factors: impact on digoxin measurements and potential physiological implications.  

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Various laboratories have reported endogenous digoxin-like immunoreactive factor(s) (DLIF) in blood from patients in renal failure or liver failure, from newborn infants, and from third-trimester pregnant women. Similar immunoreactivity has been detected in amniotic fluid, in cord blood, and in urine and serum from normal subjects. The factor(s) giving rise to this immunoreactivity cross react with antibodies used in many currently available immunoassays for digoxin, sometimes causing apparent digoxin concentrations exceeding the therapeutic range obtained for exogenous digoxin, with consequent errors in measurement and in subsequent clinical interpretation of digoxin results. Here, I summarize findings in our laboratory and those of others. DLIF evidently exist in three states in serum: tightly protein-bound, weakly protein-bound, and unbound (free). In normal subjects, greater than 90% of the total DLIF in serum is tightly but reversibly bound to serum proteins and is not readily detectable by direct measurement of digoxin in serum with conventional immunoassays. However, there seems to be a redistribution of the more weakly bound and unbound components in patients with renal failure, pregnant women, and newborns. The increased values detected in these groups are ascribable to increased amounts of weakly bound and unbound DLIF rather than to increased total DLIF. Carrier proteins may play a prominent role in the transport of these factors in blood. I discuss the potential physiological and pharmacological implications of detecting endogenous immunoreactive factors that cross react with antibodies to drugs. PMID:3896570

Valdes, R

1985-09-01

260

Influence of resting tension on immunoreactive atrial natriuretic peptide secretion by rat atria superfused in vitro  

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Atrial natriuretic peptide is a potent diuretic hormone secreted by the atria in response to volume expansion. We examined the effect of resting tension on atrial natriuretic peptide secretion by rat atria superfused in vitro. Left atria were hooked between an electrode and force transducer and superfused with medium 199. The atria were studied at a pacing frequency of 0 or 3 Hz. Atrial natriuretic peptide content of the superfusate was measured by radioimmunoassay. In nonpaced and paced atria, increasing resting tension three- to five-fold caused immunoreactive atrial natriuretic peptide secretion to increase by 35 +/- 5% (mean +/- SEM, n = 6, p less than 0.01) and 30 +/- 3% (n = 4, p less than 0.01), respectively. Lowering resting tension by 50% in nonpaced and paced atria lowered immunoreactive atrial natriuretic peptide secretion by 30 +/- 3% (n = 7, p less than 0.01) and 24 +/- 3% (n = 6, p less than 0.01), respectively. To exclude the possibility that release of norepinephrine or acetylcholine from endogenous nerve endings was mediating this effect, the atria were superfused with the combination of propranolol 0.1 microM, phentolamine 1.0 microM, and atropine 10 microM. These concentrations of the antagonists were 125-fold or higher than their Kd for binding to their respective receptors. The antagonists did not block the rise in immunoreactive atrial natriuretic peptide secretion; neither did they inhibit an established rise in immunoreactive atrial natriuretic peptide secretion induced by increasing the resting tension.

Schiebinger, R.J.; Linden, J.

1986-07-01

 
 
 
 
261

Glial fibrillary acidic protein immunoreactivity in the rat suprachiasmatic nucleus: circadian changes and their seasonal dependence  

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The pacemaker of the biological clock, the suprachiasmatic nucleus (SCN) of the hypothalamus, was studied in intact male rats to determine its immunoreactivity to glial fibrillary acidic protein (GFAP), a specific marker of astrocytes. Animals were kept under 12-h light–dark cycles in synchrony with day–night periods. Immunohistochemical reactions were carried out at midday and late at night in both winter (January) and summer (July). In winter, GFAP immunoreactivity was found to be low during the day and high at night. The findings were reversed in summer, when GFAP immunoreactivity was high during the day and low at night. Increased GFAP immunoreactivity appeared in the form of an abundance of thick immunopositive fibres rather than of cell bodies. This was interpreted as a hypertrophy of pre-existing astrocytes due to alternating photic stimulation conveyed by retinofugal fibres to the SCN. The observed seasonal reversal in the direction of GFAP oscillations raises the possibility that a circannual timer exists outside the SCN.

Gerics, Balazs; Szalay, Ferenc; Hajos, Ferenc

2006-01-01

262

Reproduction-associated immunoreactive peptides in the nervous systems of prosobranch gastropods.  

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Antibodies against reproductive peptides of Aplysia and Lymnaea were used to localize homologous immunoreactive peptides in the nervous systems of three prosobranch species: Busycon canaliculatum, Concholepas concholepas, and Tegula atra. Positive control experiments in L. stagnalis demonstrated the broad species range of the anti-egg-laying hormone (anti-ELH) antibody used in this study, and showed binding of anti-alpha-caudodorsal-cell peptide (anti-alpha-CDCP) to the same cells in cerebral and buccal ganglia. Dot immunoassays with synthetic ELH confirmed the reactivity and sensitivity (< 0.1 microgram) of the anti-ELH antibody. Experiments with preadsorbed antibody or no primary antibody confirmed its specificity. In B. canaliculatum, clusters of more than 300 neuronal cell bodies immunoreactive to both anti-ELH and anti-alpha-CDCP were observed along the medial margins of left and right cerebral ganglia. Anti-alpha-CDCP reacted with additional small populations of cerebral ganglion neurons not stained by anti-ELH. Anti-ELH and anti-alpha-CDCP also reacted with overlapping but different small populations of neurons in buccal ganglia. In C. concholepas and T atra, ELH-like immunoreactivity was found in cerebral ganglia, and in T. atra in fibers in the cerebral ganglia and cerebral-pedal connectives. Thus, cerebral ganglia are the major locus of the ELH-like immunoreactivity in prosobranchs. PMID:9924774

Ram, J L; Gallardo, C S; Ram, M L; Croll, R P

1998-12-01

263

Immunoreactive glandular kallikrein in rat plasma: a radioimmunoassay for its determination  

International Nuclear Information System (INIS)

A radioimmunoassay (RIA) has been developed to measure immunoreactive glandular kallikrein in rat plasma. To prevent the binding of radioactive kallikrein to plasma inhibitors, 125I-kallikrein was inactivated with phenylmethylsulfonyl fluoride (PMSF), a procedure that maintained 125I-kallikrein immunoreactivity. Different volumes of plasma displaced 125I-PMSF-kallikrein in a parallel fashion to the kallikrein standard curve. The sensitivity of the RIA was 200 pg, and the recovery of nonradioactive active kallikrein added to plasma was 58.7%. The concentration of immunoreactive glandular kallikrein in normal rat plasma averaged 47.1 +/- 1.7 (SE) ng/ml. Bilateral nephrectomy caused a threefold increase in circulating glandular kallikrein (50 +/- 2.7 to 167 +/- 7 ng/ml; P 125I-PMSF-kallikrein as tracer prevents the interference in the RIA caused by plasma protease inhibitors. It also indicates that the submandibular gland is an important source of the immunoreactive glandular kallikrein in rat plasma and that the kidney probably participates in its metabolism. Glandular kallikrein released by the submandibular gland into the circulation may participate in regulating local blood flow before it is inactivated by plasma inhibitors

1982-01-01

264

Developmental changes in vasoactive intestinal polypeptide immunoreactivity in the human paravertebral ganglia.  

Science.gov (United States)

Vasoactive intestinal polypeptide (VIP) belongs to the glucagon-secretin family of polypeptides and possesses numerous functions. Its existence in the mammalian central and peripheral nervous system has been widely documented. However, there are no reports on the developmental aspects of VIP-like immunoreactivity (VIP-IR) in the human postganglionic sympathetic neurons. In this study the availability and distribution of vasoactive intestinal polypeptide has been localized in human stellate ganglia neurons and nerve fibers from neonates, children and adults using the immunohistochemical method. In neonatal ganglia VIP-immunoreactive postganglionic neurons were revealed in a marked population compared to others age-groups. These nerve cells are both small and large in size and are distributed in small clusters or singly in the area of ganglia sections. In children, VIP-IR in ganglionic neurons decreases. In adult stellate ganglia, VIP-immunoreactive postganglionic neurons rarely occur. In ganglia of an individual human only varicosities of VIP-positive nerve fibers were observed. These results provide the age-dependent reduction of VIP-like immunoreactivity in human stellate ganglia neurons and suggest the different role of this peptide in the function of sympathetic ganglia neurons with age. PMID:10609054

Roudenok, V; Kühnel, W; Rogov, Y; Nerovnja, A

1999-12-01

265

Amyotrophic lateral sclerosis mutant vesicle-associated membrane protein-associated protein-B transgenic mice develop TAR-DNA-binding protein-43 pathology.  

LENUS (Irish Health Repository)

Cytoplasmic ubiquitin-positive inclusions containing TAR-DNA-binding protein-43 (TDP-43) within motor neurons are the hallmark pathology of sporadic amyotrophic lateral sclerosis (ALS). TDP-43 is a nuclear protein and the mechanisms by which it becomes mislocalized and aggregated in ALS are not properly understood. A mutation in the vesicle-associated membrane protein-associated protein-B (VAPB) involving a proline to serine substitution at position 56 (VAPBP56S) is the cause of familial ALS type-8. To gain insight into the molecular mechanisms by which VAPBP56S induces disease, we created transgenic mice that express either wild-type VAPB (VAPBwt) or VAPBP56S in the nervous system. Analyses of both sets of mice revealed no overt motor phenotype nor alterations in survival. However, VAPBP56S but not VAPBwt transgenic mice develop cytoplasmic TDP-43 accumulations within spinal cord motor neurons that were first detected at 18 months of age. Our results suggest a link between abnormal VAPBP56S function and TDP-43 mislocalization.

Tudor, E L

2010-05-19

266

Direct reticular projections of trigeminal sensory fibers immunoreactive to CGRP: potential monosynaptic somatoautonomic projections  

Science.gov (United States)

Few trigeminal sensory fibers project centrally beyond the trigeminal sensory complex, with only projections of fibers carried in its sensory anterior ethmoidal (AEN) and intraoral nerves described. Fibers of the AEN project into the brainstem reticular formation where immunoreactivity against substance P and CGRP are found. We investigated whether the source of these peptides could be from trigeminal ganglion neurons by performing unilateral rhizotomies of the trigeminal root and looking for absence of label. After an 8–14 days survival, substance P immunoreactivity in the trigeminal sensory complex was diminished, but we could not conclude that the sole source of this peptide in the lateral parabrachial area and lateral reticular formation arises from primary afferent fibers. Immunoreactivity to CGRP after rhizotomy however was greatly diminished in the trigeminal sensory complex, confirming the observations of others. Moreover, CGRP immunoreactivity was nearly eliminated in fibers in the lateral parabrachial area, the caudal ventrolateral medulla, both the peri-ambiguus and ventral parts of the rostral ventrolateral medulla, in the external formation of the nucleus ambiguus, and diminished in the caudal pressor area. The nearly complete elimination of CGRP in the lateral reticular formation after rhizotomy suggests this peptide is carried in primary afferent fibers. Moreover, the arborization of CGRP immunoreactive fibers in these areas mimics that of direct projections from the AEN. Since electrical stimulation of the AEN induces cardiorespiratory adjustments including an apnea, peripheral vasoconstriction, and bradycardia similar to those seen in the mammalian diving response, we suggest these perturbations of autonomic behavior are enhanced by direct somatic primary afferent projections to these reticular neurons. We believe this to be first description of potential direct somatoautonomic projections to brainstem neurons regulating autonomic activity.

Panneton, W. Michael; Gan, Qi

2014-01-01

267

Oxaliplatin-induced loss of phosphorylated heavy neurofilament subunit neuronal immunoreactivity in rat DRG tissue  

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Full Text Available Abstract Background Oxaliplatin and related chemotherapeutic drugs cause painful chronic peripheral neuropathies in cancer patients. We investigated changes in neuronal size profiles and neurofilament immunoreactivity in L5 dorsal root ganglion (DRG tissue of adult female Wistar rats after multiple-dose treatment with oxaliplatin, cisplatin, carboplatin or paclitaxel. Results After treatment with oxaliplatin, phosphorylated neurofilament heavy subunit (pNF-H immunoreactivity was reduced in neuronal cell bodies, but unchanged in nerve fibres, of the L5 DRG. Morphometric analysis confirmed significant changes in the number (-75%; P P P = 0.82, NF-M (-1%, P = 0.96 or NF-H (0%; P = 0.93 after oxaliplatin treatment, although the sizes of parvalbumin (-29%, P = 0.047, NF-M (-11%, P = 0.038 and NF-H (-28%; P = 0.0033 immunoreactive neurons were reduced. In an independent comparison of different chemotherapeutic agents, the number of pNF-H-immunoreactive neurons was significantly altered by oxaliplatin (-77.2%; P P = 0.03 but not by carboplatin or paclitaxel, and their mean cell body area was significantly changed by oxaliplatin (-31.1%; P = 0.008 but not by cisplatin, carboplatin or paclitaxel. Conclusion This study has demonstrated a specific pattern of loss of pNF-H immunoreactivity in rat DRG tissue that corresponds with the relative neurotoxicity of oxaliplatin, cisplatin and carboplatin. Loss of pNF-H may be mechanistically linked to oxaliplatin-induced neuronal atrophy, and serves as a readily measureable endpoint of its neurotoxicity in the rat model.

Connor Bronwen

2009-11-01

268

Atrial natriuretic factor and digoxin-like immunoreactive factor in diabetic patients: their interrelation and the influence of the autonomic nervous system.  

Science.gov (United States)

In diabetic patients, several factors contribute to volume expansion and have to be counteracted by humoral and neuronal feedback control systems. We investigated N-terminal proatrial natriuretic factor (ANF1-98) and digoxin-like immunoreactive factor (DLIF), which are two counteracting hormones, and their interrelationship, with additional consideration given to autonomic nervous function in diabetic patients. ANF1-98 and DLIF were measured in 64 diabetic patients. Autonomic nervous function was assessed using nine autonomic nervous function tests. The patients were subdivided into two groups, one with four or more (group 1) and one with less than four abnormal results in autonomic function tests (group 2). Compared with group 2, group 1 demonstrated detectable DLIF levels less often (17.2 vs. 45.7, P = 0.0195) and increased levels of ANF1-98 (mean +/- SEM: 850.0 +/- 108.8 vs. 554.8 +/- 45.9 pmol/L, P = 0.0099). However, the groups did not differ in blood pressure, daily sodium, and daily potassium excretion. The number of abnormal autonomic function tests correlated significantly with ANF1-98 (P = 0.0002). In patients with detectable DLIF, DLIF correlated with ANF1-98 (P = 0.0080). These results demonstrate close interactions between the autonomic nervous system and the two natriuretic hormones. In patients with autonomic nervous dysfunction, higher levels of ANF may possibly compensate for the lack of the natriuretic DLIF to counteract hypertension and chronic volume expansion. PMID:8784101

Straub, R H; Hall, C; Krämer, B K; Elbracht, R; Palitzsch, K D; Lang, B; Schölmerich, J

1996-09-01

269

Chromosomal phenotypes and submicroscopic abnormalities  

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Abstract The finding, during the last decade, that several common, clinically delineated syndromes are caused by submicroscopic deletions or, more rarely, by duplications, has provided a powerful tool in the annotation of the human genome. Since most microdeletion/microduplication syndromes are defined by a common deleted/duplicated region, abnormal dosage of genes located within these regions can explain the phenotypic similarities among individuals with a specific syndrome. As suc...

Devriendt Koen; Vermeesch Joris R

2004-01-01

270

Is Dark Energy Abnormally Weighting?  

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We present a new interpretation of dark energy in terms of an \\textit{Abnormally Weighting Energy} (AWE). This means that dark energy does not couple to gravitation in the same way as ordinary matter, yielding a violation of the weak and strong equivalence principles on cosmological scales. The resulting cosmological mechanism accounts for the Hubble diagram of type Ia supernovae in terms of both cosmic acceleration and variation of the gravitational constant while still acc...

Fuzfa, A.; Alimi, J. -m

2006-01-01

271

Imaging of pediatric mesenteric abnormalities  

International Nuclear Information System (INIS)

The relative paucity of mesenteric fat seen in the pediatric population can make detection and localization of processes in the mesentery difficult. This pictorial essay reviews pediatric mesenteric disorders and presents criteria that help localize processes to the mesentery. Disorders are categorized by specific patterns of involvement, which can readily be identified by imaging: developmental abnormalities of mesenteric rotation, diffuse mesenteric processes, focal mesenteric masses, and multifocal mesenteric masses. (orig.)

1999-09-01

272

Imaging of pediatric mesenteric abnormalities  

Energy Technology Data Exchange (ETDEWEB)

The relative paucity of mesenteric fat seen in the pediatric population can make detection and localization of processes in the mesentery difficult. This pictorial essay reviews pediatric mesenteric disorders and presents criteria that help localize processes to the mesentery. Disorders are categorized by specific patterns of involvement, which can readily be identified by imaging: developmental abnormalities of mesenteric rotation, diffuse mesenteric processes, focal mesenteric masses, and multifocal mesenteric masses. (orig.) With 19 figs., 24 refs.

Zarewych, Z.M.; Frush, D.P.; Bisset, G.S. III [Department of Radiology, Division of Pediatric Radiology, Duke University Medical Center, Durham, NC (United States); Donnelly, L.F. [Department of Radiology, Division of Pediatric Radiology, Duke University Medical Center, Durham, NC (United States)]|[Department of Radiology, Division of Pediatric Radiology, Durham, NC (United States)

1999-09-01

273

Computed tomography abnormalities in hanging  

International Nuclear Information System (INIS)

The CT pattern of bilateral and symmetrical round low density areas in the globi pallidi has been observed in a young man who attempted suicide by hanging. These CT abnormalities are similar to those described in other conditions such as carbon monoxide, hydrogen sulfide, cyanide and methanol poisoning, hypoglycaemia, drowning and acute global central nervous system hypoperfusion.The findings appear to be correlated with acute cerebral hypoxia. (orig.)

1987-01-01

274

Mastoid abnormalities in Down syndrome  

International Nuclear Information System (INIS)

Hearing loss and otitis media are commonly associated with Down syndrome. Hypoplasia of the mastoids is seen in many affected children and sclerosis of mastoid bones is not uncommon in Down syndrome. Awareness and early recognition of mastoid abnormality may lead to appropriate and timely therapy, thereby preserving the child's hearing or compensating for hearing loss; factors which are important for learning and maximum development. (orig.)

1989-01-01

275

Mastoid abnormalities in Down syndrome  

Energy Technology Data Exchange (ETDEWEB)

Hearing loss and otitis media are commonly associated with Down syndrome. Hypoplasia of the mastoids is seen in many affected children and sclerosis of mastoid bones is not uncommon in Down syndrome. Awareness and early recognition of mastoid abnormality may lead to appropriate and timely therapy, thereby preserving the child's hearing or compensating for hearing loss; factors which are important for learning and maximum development.

Glass, R.B.J.; Yousefzadeh, D.K.; Roizen, N.J.

1989-06-01

276

Characteristics of abnormal glow discharges  

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Here the authors report the work done in case of abnormal glow discharge in air at low pressures. The breakdown voltage measurements and I-V characteristics are obtained to find the region and the partial discharge conditions corresponding to abnormal glow discharge which also tries to occupy the maximum possible volume of the chamber. The charge density, electron and ion temperature and floating potential measurements are carried out using Langmur probe along with breakdown current pulses for understanding different regions of glow discharges. The floating potential measurements show fluctuations up to 7% level near the anode and the level decreases as one goes away from the anode in positive column of the discharge. These fluctuations are related to the ion acoustic oscillations in positive glow region. The authors could obtain the maximum charge density of 10{sup 13} cm{sup {minus}3} with ion temperature in the range of 7--9 eV. It is expected that these results can help in understanding the behavior of abnormal glow discharge which shows different characteristics under different experimental conditions and has various applications.

Kulkarni, S.V.; Panchali, D.; Kumari, V.; Bora, D. [Inst. for Plasma Research, Gujarat (India)

1996-12-31

277

Enteroendocrine profile of ?-transducin immunoreactive cells in the gastrointestinal tract of the European sea bass (Dicentrarchus labrax).  

Science.gov (United States)

In vertebrates, chemosensitivity of nutrients occurs through the activation of taste receptors coupled with G-protein subunits, including ?-transducin (G(?tran)) and ?-gustducin (G(?gust)). This study was aimed at characterising the cells expressing G(?tran) immunoreactivity throughout the mucosa of the sea bass gastrointestinal tract. G(?tran) immunoreactive cells were mainly found in the stomach, and a lower number of immunopositive cells were detected in the intestine. Some G(?tran) immunoreactive cells in the stomach contained G(?gust) immunoreactivity. Gastric G(?tran) immunoreactive cells co-expressed ghrelin, obestatin and 5-hydroxytryptamine immunoreactivity. In contrast, G(?tran) immunopositive cells did not contain somatostatin, gastrin/cholecystokinin, glucagon-like peptide-1, substance P or calcitonin gene-related peptide immunoreactivity in any investigated segments of the sea bass gastrointestinal tract. Specificity of G(?tran) and G(?gust) antisera was determined by Western blot analysis, which identified two bands at the theoretical molecular weight of ~45 and ~40 kDa, respectively, in sea bass gut tissue as well as in positive tissue, and by immunoblocking with the respective peptide, which prevented immunostaining. The results of the present study provide a molecular and morphological basis for a role of taste-related molecules in chemosensing in the sea bass gastrointestinal tract. PMID:23748963

Latorre, Rocco; Mazzoni, Maurizio; De Giorgio, Roberto; Vallorani, Claudia; Bonaldo, Alessio; Gatta, Pier Paolo; Corinaldesi, Roberto; Ruggeri, Eugenio; Bernardini, Chiara; Chiocchetti, Roberto; Sternini, Catia; Clavenzani, Paolo

2013-12-01

278

Responses of plasma cyclic AMP, serum immunoreactive insulin, C-peptide immunoreactivity and blood sugar levels to glucagon in patients with liver diseases.  

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Full Text Available Levels of plasma cyclic AMP, serum immunoreactive insulin (IRI, serum c-peptide immunoreactivity (CPR and blood sugar (BS were determined 0, 15, 30, 45 and 60 min after a glucagon injection (0.01 mg per kg body weight in normal controls, patients with acute hepatitis and liver cirrhosis. Plasma cyclic AMP responses to glucagon in liver disease patients varied widely in peak value, and only in patients with fulminant hepatitis and decompensated liver cirrhosis with poor prognosis was the response suppressed. The peak response of BS was found significantly later in liver cirrhosis patients than in normal controls. IRI and CPR responses to glucagon were lower in acute hepatitis patients than in normal controls and liver cirrhosis patients. IRI levels and their sum were also lower in acute hepatitis patients, although CPR levels were not significantly different. Thus, the ratio of the sum of CPR from 0 to 60 min to that of IRI was significantly higher in acute hepatitis, indicating impaired pancreatic secretion of insulin to glucagon stimulation as well as increased uptake of insulin by the liver in acute hepatitis.

Shimamura,Junnosuke

1985-10-01

279

Sodium channel Nav1.8 immunoreactivity in painful human dental pulp  

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Full Text Available Abstract Background The tetrodotoxin-resistant voltage-gated sodium channel Nav1.8 (SNS1/PN3 is expressed by nociceptors and may play a role in pain states. Methods Using specific antibodies for immunohistochemistry, we studied Nav1.8 – immunoreactivity in human dental pulp in relation to the neuronal marker neurofilament. Human tooth pulp was extracted from teeth harvested from a total of twenty-two patients (fourteen without dental pain, eight patients with dental pain. Results Fibres immunoreactive for Nav1.8, were significantly increased on image analysis in the painful group: median (range Nav1.8 to Neurofilament % area ratio, non-painful 0.059 (0.006–0.24, painful 0.265 (0.13–0.5, P = 0.0019. Conclusion Nav1.8 sodium channels may thus represent a therapeutic target in trigeminal nerve pain states.

Tate S

2005-07-01

280

Ablation of prion protein immunoreactivity by heating in saturated calcium hydroxide  

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Full Text Available Abstract Background Prions, the infectious agents that cause transmissible spongiform encephalopathies (TSEs, are relatively resistant to destruction by physical, enzymatic, and chemical treatments. Hydrolysis in boiling saturated calcium hydroxide (limewater utilizes inexpensive chemicals to digest protein components of offal. The purpose of this work was to determine if incubating brain material from scrapie-infected sheep in near-boiling saturated calcium hydroxide solution (Ca(OH2 would abolish immunoreactivity of the infectious prion (PrPSc as determined by western blot. Findings After incubating for as few as 10 minutes in saturated calcium hydroxide at 99°C, immunoreactivity of protease resistant bands by western blot analysis is completely lost. Conclusion Boiling in limewater may offer an alternative for disposal of carcasses and enable alternative uses for rendered products from potentially infected carcasses.

Holtzapple Mark T

2008-10-01

 
 
 
 
281

Digoxinlike immunoreactive factor isolated from human pleural fluid is structurally similar to digoxin.  

Science.gov (United States)

To further define the chemical structure of human endogenous digoxinlike immunoreactive factors (DLIF) we used human pleural effusions as a source of the substance. Digoxinlike immunoreactive factor activity was detected by radioimmunoassay in the pleural fluid of each of four patients; average concentration was 0.35 ng/mL. The chemical profile of DLIF was determined by initial extraction and concentration of DLIF by ion exchange chromatography followed by reverse phase-high-pressure liquid chromatography (RP-HPLC) separation and purification. Using high-pressure liquid chromatography cochromatography of DLIF, together with several radioactively marked glycosides, we observed a single peak of DLIF activity that was chromatographically identical to digoxin. The present study further supports the recent finding that DLIF is related structurally to the cardiac glycosides, and for the first time it has been proven that DLIF is present in pleural fluids. PMID:9216437

Weinberg, U; Dolev, S; Shapiro, M S; Shilo, L; Rabinowitz, R; Shenkman, L

1997-07-01

282

MR imaging of abnormal synovial processes  

International Nuclear Information System (INIS)

MR imaging can directly image abnormal synovium. The authors reviewed over 50 cases with abnormal synovial processes. The abnormalities include Baker cysts, semimembranous bursitis, chronic shoulder bursitis, peroneal tendon ganglion cyst, periarticular abscesses, thickened synovium from rheumatoid and septic arthritis, and synovial hypertrophy secondary to Legg-Calve-Perthes disease. MR imaging has proved invaluable in identifying abnormal synovium, defining the extent and, to a limited degree, characterizing its makeup

1987-12-04

283

Laminar and subcellular heterogeneity of GLAST and GLT-1 immunoreactivity in the developing postnatal mouse hippocampus.  

Science.gov (United States)

Astrocytes express two sodium-coupled transporters, glutamate-aspartate transporter (GLAST) and glutamate transporter-1 (GLT-1), which are essential for the maintenance of low extracellular glutamate levels. We performed a comparative analysis of the laminar and subcellular expression profile of GLAST and GLT-1 in the developing postnatal mouse hippocampus by using immunohistochemistry and western blotting and employing high-resolution fluorescence microscopy. Astrocytes were identified by costaining with glial fibrillary acidic protein (GFAP) or S100?. In CA1, the density of GFAP-positive cells and GFAP expression rose during the first 2 weeks after birth, paralleled by a steady increase in GLAST immunoreactivity and protein content. Upregulation of GLT-1 was completed only at postnatal days (P) P20-25 and was thus delayed by about 10 days. GLAST staining was highest along the stratum pyramidale and was especially prominent in astrocytes at P3-5. GLAST immunoreactivity indicated no preferential localization to a specific cellular compartment. GLT-1 exhibited a laminar expression pattern from P10-15 on, with the highest immunoreactivity in the stratum lacunosum-moleculare. At the cellular level, GLT-1 immunoreactivity did not entirely cover astrocyte somata and exhibited clusters at processes. In neonatal and juvenile animals, discrete clusters of GLT-1 were also detected at perivascular endfeet. From these results, we conclude there is a remarkable subcellular heterogeneity of GLAST and GLT-1 expression in the developing hippocampus. The clustering of GLT-1 at astrocyte endfeet indicates that it might serve a specialized functional role at the blood-brain barrier during formation of the hippocampal network. PMID:23939750

Schreiner, Alexandra E; Durry, Simone; Aida, Tomomi; Stock, Martin C; Rüther, Ulrich; Tanaka, Kohichi; Rose, Christine R; Kafitz, Karl W

2014-01-01

284

Determination of immunoreactivity ofPlasmodium falciparum antigens, serum dilutions and biomaterials  

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Immunoreactivity properties of serum dilutions andPlasmodium falciparum malaria antigens were measured and compared by ELISA technique using different ELISA plates to evaluate the role of antigens and serum dilutions for optimum binding. Also effort has been made to see the effect of reaction surface and material i.e. ELISA plates for binding capacity. Serological properties were estimated by ELISA methods for detection of malaria and determination of immunological characteristics. Three Pf a...

2004-01-01

285

Isolation and morphology of an immunoreactive outer wall fraction produced by spherules of Coccidioides immitis.  

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A previously undescribed, immunoreactive, membranous spherule outer wall (SOW) fraction produced by Coccidioides immitis (strains 634 and 735) grown in culture was isolated. Both this fraction and intact spherules were reactive with sera from coccidioidomycosis patients, as demonstrated by immunofluorescence microscopy. The serological activity of SOW was also demonstrated by its reactivity with human anti-C. immitis tube precipitin in a standardized immunodiffusion assay. Extraction of SOW w...

1988-01-01

286

Nesfatin-1 immunoreactivity in rat brain and spinal cord autonomic nuclei  

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Nesfatin-1 is one of the peptide products of posttranslational processing of the nucleobindin-2 (NUCB2) gene, suggested to have physiological relevance to suppress food intake and body weight gain in rats. Nesfatin-1-immunoreactive cells have been found in distinct nuclei in the rat brain related to circuitries regulating food intake. Here, we report novel yet undescribed localization of NUCB2/nesfatin-1 at the mRNA and protein level in the rat central nervous system. Immunohistochemical stai...

Goebel, Miriam; Stengel, Andreas; Lambrecht, Nils W. G.; Wang, Lixin; Tache?, Yvette

2009-01-01

287

Immunoreactive forms of cationic trypsin in plasma and ascitic fluid of dogs in experimental pancreatitis.  

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A canine model of bile-induced pancreatitis has been employed to investigate time-dependent changes in the molecular forms of trypsin in blood and ascitic fluid in this disease. The distribution of immunoreactive trypsin as trypsinogen and trypsin bound to plasma inhibitors in ascitic fluid and plasma during the course of the disease has been investigated by means of a radioimmunoassay for canine pancreatic cationic trypsin. In addition, trypsinlike amidase activity was determined in plasma a...

1981-01-01

288

Independent prognostic value of fascin immunoreactivity in stage III–IV colonic adenocarcinoma  

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Fascin, an actin-bundling protein involved in cell motility, has been shown to be upregulated in several types of carcinomas. In this study, we investigated the expression of fascin in 228 advanced colonic adenocarcinoma patients with a long follow-up. Fascin expression was compared with several clinicopathologic parameters and survival. Overall, fascin immunoreactivity was detected in 162 (71%) tumours with a prevalence for right-sided tumours (P<0.001). Fascin correlated significantly with ...

Puppa, G.; Maisonneuve, P.; Sonzogni, A.; Masullo, M.; Chiappa, A.; Valerio, M.; Zampino, M. G.; Franceschetti, I.; Capelli, P.; Chilosi, M.; Menestrina, F.; Viale, G.; Pelosi, G.

2007-01-01

289

Distribution of galanin immunoreactivity in the respiratory tract of pig, guinea pig, rat, and dog.  

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Galanin, a newly discovered peptide isolated from porcine intestine, is known to cause contraction in rat smooth muscle preparations and to induce hyperglycaemia in dogs. By the use of radioimmunoassay and immunohistochemical techniques the concentration and distribution of galanin immunoreactivity were determined in several areas of the respiratory tract of five dogs, five guinea pigs, five rats, and two pigs. Antibodies were raised in rabbits to whole unconjugated natural porcine galanin. T...

1985-01-01

290

Immunoreactivity of Glutamate in Mouse Retina Inner Segment of Photoreceptors With In Vivo Cryotechnique  

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The purpose of this study was to clarify a previously controversial issue concerning glutamate (Glu) immunoreactivity (IR) in the inner segment (IS) of photoreceptors by using in vivo cryotechnique (IVCT) followed by freeze substitution (FS), which enabled us to analyze the cells and tissues reflecting living states. Eyeballs from anesthetized mice were directly frozen using IVCT. The frozen tissues were processed for FS fixation in acetone containing chemical fixatives, and embedded in paraf...

Terada, Nobuo; Ohno, Nobuhiko; Saitoh, Sei; Saitoh, Yurika; Ohno, Shinichi

2009-01-01

291

Distribution of Glucagon-like Peptide-1 Immunoreactivity in the Hypothalamic Paraventricular and Supraoptic Nuclei  

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Glucagon-like peptide-1 (GLP-1) plays a role in modulating neuroendocrine and autonomic function. The hypothalamic paraventricular nucleus (PVN) contains aggregations of GLP-1 fibers and expresses GLP-1 receptors, making it a likely site of action for GLP-1 signaling. The current study was designed to establish domains of GLP-1 action, focusing on axosomatic appositions on different neuroendocrine and autonomic cell populations in the PVN. The data indicate abundant GLP-1-immunoreactive termi...

Tauchi, Miyuki; Zhang, Rong; D’alessio, David A.; Stern, Javier E.; Herman, James P.

2008-01-01

292

Human seminal alpha-inhibins: detection in human pituitary, hypothalamus, and serum by immunoreactivity.  

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An antiserum generated in rabbits against human seminal alpha-inhibin-52 has been used to develop a sensitive radioimmunoassay for the detection of alpha-inhibins. The alpha-inhibin-52 antiserum reacts with alpha-inhibin-92 and alpha-inhibin-31 with equal avidity. These peptides were found to be present in human pituitary, hypothalamus, and serum. In exclusion chromatography on Sephadex G-100, the immunoreactive material eluted in a large molecular size region. Immunoblot analysis of column-d...

Ramasharma, K.; Li, C. H.

1986-01-01

293

Chromosome abnormality in early entrants  

International Nuclear Information System (INIS)

A preliminary study was performed to estimate radiation dose in 10 individuals who had entered Hiroshima city one day after the explosion and worked around the hypocenter area for a week. The numbers of cells with chromosome aberrations differed in each subject, ranging from one to 18 (median: 11 abnormalities) in 500 metaphases of lymphocytes cultured for 48 hours with phytohemagglutinin. Five individuals had more than 1.6 percent stable type aberrations and were estimated to have been exposed to 6-13 rad. The other five individuals and the controls were estimated to be less than two rad. (author)

1991-01-01

294

Heterogeneity of human plasma insulin: techniques for separating immunoreactive components and their determination by radioimmunoassay  

International Nuclear Information System (INIS)

When human plasma is filtered on Sephadex G-SO fine, insulin immunoreactivity is recovered in two peaks: 'big insulin', the higher molecular weight component and 'little insulin', the lower molecular component, having elution volumes that correspond to those of porcine proinsulin 125I and porcine insulin 125I respectively. The presence of another form of immunoreactive insulin 'big big insulin' was detected from an insuloma suspect and its elution pattern corresponding to serum albumin. The eluates correspondent to 'big' and 'little' insulin as well as 'big big' component were assayed by radioimmunoassay using crystalline human insulin as a standard, porcine insulin 125 tracer and anti insulin serum. The antibody, raised in guinea-pigs, was sensitive and potent being adequate for the assay. The reactivity of insulin and proinsulin was tested against the antibody. The relative proportions of several components of total immunoreactive insulin in plasma were studied in basal conditions in five normal subjects and in the patient JSC with pancreatic insulin-secreting tumor as well as after glucose stimuli in all tolbutamide in JSC. (author)

1977-01-01

295

Inducible nitric oxide synthase immunoreactivity in the granulomatous intestinal lesions of naturally occurring bovine Johne's disease.  

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Inducible nitric oxide synthase (iNOS) is important in the control of a number of intracellular pathogens, including mycobacteria, and is a marker of classic macrophage activation. In human granulomatous diseases such as leprosy, a spectrum of granulomatous lesions is described, ranging from the tuberculoid to lepromatous types. Tuberculoid granulomas are associated with enhanced iNOS production and improved clinical outcomes over the lepromatous types. The aim of this study is to determine whether an association exists between morphology of bovine Johne's disease granulomas and lesion macrophage effector functions. To accomplish this, we retrospectively evaluated 24 cases of bovine Johne's disease. In each case, we recorded the predominant granuloma morphology and evaluated iNOS immunoreactivity and bacterial burden by acid-fast stains and mycobacterial immunolabeling. The results of this study demonstrate that all cases had granulomas with features most similar to the lepromatous type. This morphology correlated with heavy bacterial burdens demonstrated by acid-fast staining and mycobacterial immunoreactivity. None of the cases had high expression of iNOS in mycobacterial-positive granulomas. When iNOS immunoreactivity was identified, it was usually located near the crypts and was distinct from the granulomatous foci. PMID:15872370

Hostetter, J; Huffman, E; Byl, K; Steadham, E

2005-05-01

296

Distribution of gastrin-releasing peptide immunoreactivity in the brain of the collared dove (Streptopelia decaocto).  

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The distribution of immunoreactivity after applying an antibody against gastrin-releasing peptide (GRP) was studied in the brain of the collared dove (Streptopelia decaocto). In the forebrain GRP-immunoreactive (GRP-ir) cells were found in the hyperstriatum accessorium, medial and lateral parts of the neostriatum, corticoidea dorsolateralis and temporoparieto-occipitalis areas, hippocampus, pre- and parahippocampal areas and prepiriform cortex. In the brainstem, GRP-ir cells were restricted mainly to the substantia nigra and ventral tegmental nucleus. Areas with densely packed GRP-ir clusters of varicosities were the medial intermediate hyperstriatum ventrale and lateral septal nucleus; dense GRP-ir neuropil was found in the parolfactory lobe, and in the dorsal half of the intermediate and caudal archistriatum. The ventral lamina medullaris contained many GRP-ir fibers. Forebrain areas devoid of immunoreactivity were the basal nucleus, ectostriatum, rostral archistriatum, most of the paleostriatum augmentatum and the lateral bed nucleus of the stria terminalis. Moderate densities of GRP-ir elements were found in the other telencephalic areas and further in, among others, the preoptic and hypothalamic region, ventral area of Tsai, cerulean nuclei, parabrachial complex, dorsal glossopharyngeal and vagus motor nuclei and medial nuclei of the solitary complex. The observations are compared with data from the literature and the implications for the definition of specific centers within the avian brain are discussed, with emphasis on systems with a role in visceral and motivational functions and in learning. PMID:10805083

Dubbeldam, J L; den Boer-Visser, A M

2000-04-01

297

Criteria for identifying endogenous compounds as digoxin-like immunoreactive factors in humans.  

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Endogenous digoxin-like immunoreactive factors (DLIF) are factors in plasma that interact with anti-digoxin antibodies. In this report we propose specific empirical criteria that must be satisfied by any group of endogenous compounds purported to account for DLIF activity in human plasma. These criteria include immunoreactive potency relative to existing physiologic concentrations as well as the biochemical and protein binding properties of these compounds. Recent studies have identified several congeners of fatty acids and phospholipids, hydrocortisone, and dehydroepiandrosterone-sulfate as compounds likely to account for DLIF activity in plasma. Using the above criteria we demonstrate that the highest reported plasma concentrations of these compounds combined account for less than 25% of DLIF reported in healthy adult subjects, less than 11% in newborns, less than 27% in pregnant women, and less than 39% in patients with renal failure. Human serum albumin at a concentration of 40 g/l completely abolished any detectable interaction of these compounds with both anti-digoxin antibodies or canine kidney Na/K-ATPase. The immunoreactive and physical properties of these compounds are also not consistent with those reported for DLIF. We conclude that these compounds do not account for the plasma DLIF concentrations measured in human subjects nor are they likely to play a role as specific endogenous regulators of Na/K-ATPase. PMID:2844442

Lau, B W; Valdes, R

1988-06-30

298

[Soluble expression, purification and immunoreactive identification of mouse zona pellucida 3 fusion protein].  

Science.gov (United States)

Mammalian zona pellucida 3(ZP3) plays an important role in the induction of capacitating sperm acrosome reaction. In this study, we obtained the soluble mZP3 fusion protein and identified its immunoreactivity. mZP3 cDNA was cloned into plasmid pMAL-p2x, and the recombinant plasmid was transformed into Escherichia coli BL21. To get the soluble mZP3 fusion protein, we tried to optimize the expression conditions, including additives, IPTG concentrations, temperatures and induction duration. Then, Western blotting and ELISA were used to identify the immunoreactivity of the purified protein. Based on the optimization experiments, we concluded that the best soluble expression conditions for the mZP3 fusion protein involved incubation to an A600 of 0.6, addition of glucose to a final concentration of 0.02 mol/L, addition of IPTG to a final concentration of 0.6 mmol/L and then further incubation for 4 h at 25 degrees C. Western blotting and ELISA showed that the mZP3 fusion protein retained immunoreactivity. The fusion protein can be used as solubility antigens for developing the immunocontraception vaccines of mZP3 and detecting the immune effects of the vaccine. PMID:19938453

Sun, Meiyu; Ma, Zhenghai; Li, Yongxin; Lü, Tao; Chen, Kaixu; Zhang, Fuchun

2009-08-01

299

Simultaneous detection of Clavibacter michiganensis subsp. nebraskensis and Pantoea stewartii subsp. stewartii based on microsphere immunoreaction.  

Science.gov (United States)

Clavibacter michiganensis subsp. nebraskensis (Cmn) and Pantoea stewartii subsp. stewartii (Pss) are two plant pathogens that can cause tremendous agricultural economic losses. This novel method based on microsphere immunoreaction was developed for the simultaneous detection of Cmn and Pss in maize. This multiplex method was constructed based on microsphere immunodetection with fluorescent labels such as quantum dots (QDs) and R-phycoerythrin (R-PE) for the detection of Cmn and Pss. Captured QDs and R-PE serve as signal reporters for fluorescent readout. The principle of this method is based on a sandwich immunoreaction. Cmn and Pss captured by the microspheres were detected using flow cytometry. The limit of detection of this method was 10 times lower than the enzyme-linked immunosorbent assay (ELISA), and its analysis time (1 h) was much shorter compared with ELISA (6-8 h). The method, which has been proven to be an effective approach to multiplex detection of plant bacteria (Cmn and Pss as models), not only increased the varieties but also improved the sensitivity. The microsphere immunoreaction provides a universal method for the multiplex determination of microbes because of its high sensitivity, specificity, and speed. In the future, the method will be more fully validated in vivo to detect diversiform bacteria. PMID:23169888

Zhang, Fan; Li, Jinfeng; Zou, Mingqiang; Chen, Yan; Wang, Yanfei; Qi, Xiaohua

2013-04-01

300

Transient abnormal Q waves during exercise electrocardiography  

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Myocardial ischaemia during exercise electrocardiography is usually manifested by ST segment depression or elevation. Transient abnormal Q waves are rare, as Q waves indicate an old myocardial infarction. The case of a patient with exercise induced transient abnormal Q waves is reported. The potential mechanisms involved in the development of such an abnormality and its clinical implications are discussed.

Alameddine, F. F.; Zafari, A. M.

2004-01-01

 
 
 
 
301

Distribution of DARPP-32 immunoreactive structures in the quail brain: anatomical relationship with dopamine and aromatase.  

Science.gov (United States)

We recently demonstrated that dopamine (DA) as well as different DA receptor agonists and antagonists are able to decrease within a few minutes the aromatase activity (AA) measured in vitro in homogenates or in explants of the quail preoptic area - hypothalamus. In addition, DA also appears to regulate AA, in vivo presumably by modifying enzyme synthesis. The cellular mechanisms and the anatomical substrate that mediate these controls of AA by DA are poorly understood. Tyrosine hydroxylase-immunoreactive (TH-ir) fibers and punctate structures have been previously observed in close vicinity of aromatase-immunoreactive (ARO-ir) cells in the quail medial preoptic nucleus (POM) and bed nucleus striae terminalis (BST) but these fibers could reflect a noradrenergic innervation. We also do not know whether aromatase cells are dopaminoceptive. The main goal of the present study was therefore to bring more information on the anatomical relationships between aromatase expressing neurons and the dopaminergic system in the quail brain. The visualization by immunocytochemistry of DA and of the D1 receptor associated protein DARPP-32 was used to address these questions. DA-ir fibers were observed in the quail forebrain and overlapped extensively with nuclei that contain high densities of ARO-ir cells such as the POM and BST. This confirms that the previously reported TH-ir innervation of ARO-ir cells is, at least in part, of dopaminergic nature. DARPP-32-immunoreactive cells were found in periventricular position throughout the hypothalamus. DARPP-32-ir cells were also observed in telencephalic and mesencephalic areas (hyperstriatum accessorium, paleostriatum, nucleus intercollicularis, optic tectum). DARPP-32-ir fibers were widespread in tel-, di-, and mes-encephalic areas. The highest densities of immunoreactive fibers were detected in the lobus parolfactorius, paleostriatum augmentatum and substantia nigra/area ventralis of Tsai. In double-labeled sections, appositions between DARPP-32 fibers and ARO-ir cells were present in the dorsolateral POM and BST but DARPP-32 immunoreactivity was not detected in the ARO-ir perikarya (no colocalization). These data confirm the presence of a dopaminoceptive structures within the main cell clusters of ARO-ir cells in the quail brain but provide no evidence that these ARO-ir cells are themselves dopaminoceptive. Because DARPP-32 is not present in all types of cells expressing DA receptors, the presence of DA receptors that would not be associated with DARPP-32 in ARO-ir cells still remains to be investigated PMID:11173218

Absil, P; Foidart, A; Hemmings, H C; Steinbusch, H W; Ball, G F; Balthazart, J

2001-01-01

302

Comparison of alpha-synuclein immunoreactivity in the spinal cord between the adult and aged beagle dog  

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Alpha-synuclein (?-syn) is a presynaptic protein that is richly expressed in the central and peripheral nervous systems of mammals, and it is related to the pathogenesis of Parkinson's disease and other neurodegenerative disorders. In the present study, we compared the distribution of the immunoreactivity of ?-syn and its related gliosis in the spinal cord of young adult (2-3 years) and aged (10-12 years) beagle dogs. We discovered that ?-syn immunoreactivity was present in many neurons in...

Ahn, Ji-hyeon; Choi, Jung-hoon; Park, Joon-ha; Yan, Bing-chun; Kim, In-hye; Lee, Jae-chul; Lee, Dae-hwan; Kim, Jin-sang; Shin, Hyung-cheul; Won, Moo-ho

2012-01-01

303

CHROMOSOMAL ABNORMALITIES IN PATIENTS WITH RECURRENT MISCARRIAGE  

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Full Text Available Chromosomal abnormalities are involved in the etiology of recurrent spontaneous pregnancy loss and sub-fertility. The purpose of this study was to determine the frequency and contribution of chromosomal abnormalities in recurrent miscarriages. The results obtained and literature review are helpful in understanding the importance of cytogenetics analysis of female infertility. To investigate the distribution of chromosomal abnormalities in the Romanian population with recurrent miscarriage, karyotype analysis by G-banding was performed from peripheral blood in 967 women infertility. Results: Chromosomal abnormalities were found to 79 women (8,17%. The percentage of chromosomal abnormalities in the studied population correlates with the data in the literature. Chromosomal abnormalities could play the important role in etiology of infertility and are more frequently detected in this group of patients compared to general population. In the infertile couples balanced chromosomal abnormalities are the main cause of spontaneous abortions.

Veronica Stoian

2012-06-01

304

Is Dark Energy Abnormally Weighting?  

CERN Multimedia

We present a new interpretation of dark energy in terms of an \\textit{Abnormally Weighting Energy} (AWE). This means that dark energy does not couple to gravitation in the same way as ordinary matter, yielding a violation of the weak and strong equivalence principles on cosmological scales. The resulting cosmological mechanism accounts for the Hubble diagram of type Ia supernovae in terms of both cosmic acceleration and variation of the gravitational constant while still accounting for the present tests of general relativity. This explanation allows to build dark energy models (i) without violation of the strong energy condition $p<-\\rho c^2/3$ (ii) with non-negligible direct couplings to gravitation and (iii) natural convergence mechanism toward general relativity.

Füzfa, A

2006-01-01

305

ECG ABNORMALITIES IN RURAL AREAS  

Directory of Open Access Journals (Sweden)

Full Text Available A survey was carried out in the summer of 1972 in east Azerbaijan, northwest Iran, to determine the prevalence of cardiovascular disease. ECG tracings were prepared from 198 women and 178 men aged 40-60. Only 45% of the tracings were completely normal. Q/QS abnormalities were found in 4.7% of tracings, left axis deviation in 4% tall R wave in 37%, ST depression in 3.9% of men and 14.1% of women and T wave inversion in 2.2% of men and 11.6% of women. Further studies are recommended to explain this high prevalence of ST depression and T wave inversion in the women of this area.

M. Daneshapjooh

1975-08-01

306

Abnormal Iron Homeostasis and Neurodegeneration  

Directory of Open Access Journals (Sweden)

Full Text Available Abnormal iron metabolism is observed in many neurodegenerative diseases, however only two have shown dysregulation of brain iron homeostasis as the primary cause of neurodegeneration. Herein, we review one of these - hereditary ferritinopathy (HF or neuroferritinopathy, which is an autosomal dominant, adult onset degenerative disease caused by mutations in the ferritin light chain (FTL gene. HF has a clinical phenotype characterized by a progressive movement disorder, behavioral disturbances, and cognitive impairment. The main pathologic findings are cystic cavitation of the basal ganglia, the presence of ferritin inclusion bodies (IBs, and substantial iron deposition. Mutant FTL subunits have altered sequence and length but assemble into soluble 24-mers that are ultrastructurally indistinguishable from those of the wild type. Crystallography shows substantial localized disruption of the normally tiny 4-fold pores between the ferritin subunits because of unraveling of the C-termini into multiple polypeptide conformations. This structural alteration causes attenuated net iron incorporation leading to cellular iron mishandling, ferritin aggregation, and oxidative damage at physiological concentrations of iron and ascorbate. A transgenic murine model parallels several features of HF, including a progressive neurological phenotype, ferritin IB formation, and misregulation of iron metabolism. These studies provide a working hypothesis for the pathogenesis of HF by implicating (1 a loss of normal ferritin function that triggers iron accumulation and overproduction of ferritin polypeptides, and (2 a gain of a toxic function through radical production, ferritin aggregation, and oxidative stress. Importantly, the finding that ferritin aggregation can be reversed by iron chelators and oxidative damage can be inhibited by radical trapping may be used for clinical investigation. This work provides new insights into the role of abnormal iron metabolism in neurodegeneration.

RubenVidal

2013-07-01

307

Immunoreactivity of a 10-kDa antigen of Mycobacterium tuberculosis.  

Science.gov (United States)

Identification of Ag of Mycobacterium tuberculosis recognized by T cells is essential to understanding the pathogenesis of tuberculosis and mechanism(s) of resistance to infection. Previous studies evaluating the immunoreactivity of nitrocellulose transfers of M. tuberculosis Ag separated by SDS-PAGE indicated that a high proportion of M. tuberculosis-reactive T cell lines proliferate in response to a 10-kDa Ag. We therefore purified this Ag from M. tuberculosis culture filtrates and evaluated its immunoreactivity in patients with tuberculous infection. Proliferative responses of PBMC to the 10-kDa Ag were similar to those induced by whole M. tuberculosis and greater than those elicited by other proteins isolated from culture filtrate. Furthermore, in patients with tuberculous pleuritis, proliferative responses to the 10-kDa Ag were higher in pleural fluid mononuclear cells than in PBMC, indicating that T cell reactivity to this Ag is enhanced at the site of disease. The first 15 amino acids of the 10-kDa Ag were identical to those defined previously for Bacillus Calmette-Guérin-a (BCG-a), and a T cell clone recognized the 10-kDa Ag and a peptide of BCG-a, indicating that the 10-kDa Ag corresponds to BCG-a. This Ag elicited IFN-gamma production by pleural fluid mononuclear cells and by PBMC from healthy tuberculin reactors, suggesting that the 10-kDa Ag can enhance macrophage activation and resistance to mycobacterial infection. Our findings indicate that the 10-kDa Ag of M. tuberculosis is highly immunoreactive and should be evaluated for its capacity to elicit protective immunity. PMID:1371791

Barnes, P F; Mehra, V; Rivoire, B; Fong, S J; Brennan, P J; Voegtline, M S; Minden, P; Houghten, R A; Bloom, B R; Modlin, R L

1992-03-15

308

Interference from digitoxin-like immunoreactive factors reduced in a new monoclonal chemiluminescent digitoxin assay.  

Science.gov (United States)

Endogenous digoxin-like immunoreactive factors (DLIF) can interfere with some digoxin immunoassays. We looked for similar interference, called digitoxin-like immunoreactive factors (DTLIF) in two digitoxin immunoassays: A new chemiluminescent assay (CLIA), processed on the automated random access immunoassay system ACS:180, and a fluorescent polarization assay (FPIA), processed on the semiautomated TDx batch analyzer. One hundred thirty-seven samples of sera were tested from nondigitalized pregnant women, patients with liver or kidney diseases, and cord blood. The CLIA digitoxin assay uses a murine monoclonal antibody and requires no sample pretreatment; the FPIA digitoxin assay uses a polyclonal rabbit antibody and requires sample precipitation. Both assays have a similar dynamic range and sensitivity and give comparable results with commercial controls and external quality control survey samples. Although the CLIA detected no digitoxin in any sample tested, the FPIA showed apparent digitoxin concentrations of more than 2.0 ng/ml for 100% and 44% among cord blood and liver disease specimens, respectively. The highest DTLIF concentration was found in serum from a patient with liver disease (18.1 ng/ml). When spiked with 32 ng/ml digitoxin, six of the samples containing DTLIF generated FPIA digitoxin values of 6% to 27.5% more than the expected digitoxin levels. Two specimens with no detectable DTLIF activity were run as controls, and when spiked with digitoxin, showed target digitoxin concentrations in the FPIA. The CLIA recovered near the target digitoxin values (32 ng/ml) in all spiked samples. It was concluded that the polyclonal FPIA digitoxin assay may give discordant digitoxin concentrations in some patient groups because of interference from digitoxin-like immunoreactive factors. The CLIA digitoxin assay is not affected by DTLIF interference. PMID:9853984

Datta, P; Dasgupta, A

1998-12-01

309

Increased immunoreactive 11-ketotestosterone concentrations in sheep feces after acth challenge.  

Science.gov (United States)

11-Oxoetiocholanolone and related substances are important metabolites of cortisol and are excreted via feces in ruminants. To investigate whether 11-ketotestosterone (11-KT) or its immunoreactive metabolites are formed and excreted in ruminant feces, an enzyme immunoassay (EIA) was developed and validated. The antibody was raised in rabbits against 11-KT-3-CMO:bovine serum albumin with biotinylated 11-KT as a label. The assay showed a sensitivity of 0.3?pg/well. To validate the assay biologically, 6 rams were injected with a synthetic analogue of the adrenocorticotropic hormone (Synacthen, 2?µg/kg body wt). An aliquot was collected of each fecal portion spontaneously defecated 8?h before Synacthen injection to 24?h after injection and stored at -20?°C until analysis. Samples (0.5?g) were extracted using 80% methanol and immunoreactive metabolites measured using the 11-KT EIA and an already established 11,17-dioxoandrostane (11,17-DOA) EIA. High-performance liquid chromatography separation revealed no peak in the same elution position as authentic 11-KT; therefore, reacting substances were referred to as 11-KT equivalents. In the case of 11-KT immunoreactive substances, the values increased from baseline (median, 136?ng/g feces) to a peak concentration (median, 424?ng/g) 10 to 14?h after Synacthen injection and declined afterwards. Concentrations of 11,17-DOA showed the same pattern, but the values were 2 to 4 times higher. From this data, the authors conclude that 11-KT-like substances, specifically C19 O3 -androgens with a 17ß-hydroxy group, were present in the feces. These substances originate from the adrenals and are most likely cortisol metabolites. PMID:23404733

Sid-Ahmed, Omer; Arias, Nino; Palme, Rupert; Möstl, Erich

2013-06-01

310

Advantage of highly immunoreactive monoclonal antibodies in radioimmunoscintigraphy for tumor detection, (1)  

International Nuclear Information System (INIS)

Immunoreactivity (IR) is the fraction of a monoclonal antibody (MoAb) preparation capable of binding to an excess of a specific antigen. One of the most important requirements for successful radioimmunoscintigraphy is to use a highly immunoreactive MoAb. To assess the effect of an antibody IR on biodistribution, a fast and simple purification method has been developed using a high performance liquid chromatography (HPLC) system equipped with a hydroxylapatite (HA) column. The column was eluted at ambient temperature with 0.12 M sodium phosphate buffer (pH 6.8). With this system, the F ab fragments from the MoAb 96.5 against the human melanoma associated p97 antigen were separated into two well-resolved peaks at retention times of 6 and 16 min. FEM-XII cells (human skin melanoma cell line) were used in a cell binding assay (CBA) to determine the maximal percent IR and the affinity constant of each HA-HPLC peak. The second peak from an 125I-F ab 96.5 showed approximately two times greater maximal binding than did the first peak, whereas the affinity constant for the two was the same. This indicated that the F ab 96.5 preparations used in this study were a mixture of more active and less active components. Moreover, prior to the HA-HPLC experiments, these preparations were analyzed with a gel filtration HPLC showing a single molecular weight peak. This suggested that the HA-HPLC separation was not based on molecular weight differences although the separation mechanism of HA has not yet been fully understood. Thereby, it is concluded that the HA-HPLC is a powerful tool to purify MoAbs into the higher immunoreactive fraction which has a potential advantage in tumor targeting. (author)

1988-01-01

311

Lithium treatment and thyroid abnormalities  

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Full Text Available Abstract Background Although the interactions between lithium treatment and thyroid function have long been recognised, their clinical relevance is still controversial. This paper sets out a review of the literature to date, considering that lithium still represents the gold standard among prophylactic treatments of manic-depression several decades after its introduction. Method PubMed database was used to search for English-language articles relating to lithium treatment and thyroid function. As the amount of relevant papers totalled several hundreds, this review refers to previous reviews, especially with regard to older literature. Moreover, the authors particularly refer to a series of studies of thyroid function performed in a cohort of patients at different stages of lithium treatment, who were followed up by their group from 1989 onwards. Results The main findings from this review included: a lithium definitely affects thyroid function as repeatedly shown by studies on cell cultures, experimental animals, volunteers, and patients; b inhibition of thyroid hormone release is the critical mechanism in the development of hypothyroidism, goitre, and, perhaps, changes in the texture of the gland which are detected by ultrasonic scanning; c compensatory mechanisms operate and prevent the development of hypothyroidism in the majority of patients; d when additional risk factors are present, either environmental (such as iodine deficiency or intrinsic (immunogenetic background, compensatory potential may be reduced and clinically relevant consequences may derive; e hypothyroidism may develop in particular during the first years of lithium treatment, in middle-aged women, and in the presence of thyroid autoimmunity; f thyroid autoimmunity is found in excess among patients suffering from affective disorders, irrespective of lithium exposure; g in patients who have been on lithium for several years, the outcome of hypothyroidism, goitre, and thyroid autoimmunity do not much differ from those observed in the general population; h hyperthyroidism and thyroid cancer are observed rarely during lithium treatment. Recommendations Thyroid function tests (TSH, free thyroid hormones, specific antibodies, and ultrasonic scanning should be performed prior to starting lithium prophylaxis. A similar panel should be repeated at one year. Thereafter, annual measurements of TSH may be sufficient to prevent overt hypothyroidism. In the presence of raised TSH or thyroid autoimmunity, shorter intervals between assessments are advisable (4–6 months. Measurement of antibodies and ultrasonic scanning may be repeated at 2-to-3-year intervals. The patient must be referred to the endocrinologist if TSH concentrations are repeatedly abnormal, and/or goitre or nodules are detected. Thyroid function abnormalities should not constitute an outright contraindication to lithium treatment, and lithium should not be stopped if a patient develops thyroid abnormalities. Decisions should be made taking into account the evidence that lithium treatment is perhaps the only efficient means of reducing the excessive mortality which is otherwise associated with affective disorders.

Bocchetta Alberto

2006-09-01

312

Chromosomal abnormalities in child psychiatric patients.  

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To determine the frequency of chromosomal abnormalities in a child psychiatric population, and to evaluate possible associations between types of abnormalities and patient's clinical characteristics, cytogenetic examination was performed on 604 patients. Demographic data, reasons for karyotyping, clinical signs, and other patient characteristics were assessed and correlated with the results from karyotyping. Chromosomal abnormalities were found in 69 patients (11.3%); these were structural in...

Hong, K. E.; Kim, J. H.; Moon, S. Y.; Oh, S. K.

1999-01-01

313

CHROMOSOMAL ABNORMALITIES IN PATIENTS WITH RECURRENT MISCARRIAGE  

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Chromosomal abnormalities are involved in the etiology of recurrent spontaneous pregnancy loss and sub-fertility. The purpose of this study was to determine the frequency and contribution of chromosomal abnormalities in recurrent miscarriages. The results obtained and literature review are helpful in understanding the importance of cytogenetics analysis of female infertility. To investigate the distribution of chromosomal abnormalities in the Romanian population with recurrent miscarriage,...

Daniela Mierla; Viorica Radoi; Veronica Stoian

2012-01-01

314

Auditory abnormalities in children with autism  

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The present study aimed to describe the characteristics of auditory abnormalities present in cases of autism. One hundred and fifty six children with autism and 141matched controls with language delay were investigated via direct observations combined with parent/caregiver reports. All of the autistic individuals demonstrated auditory abnormalities especially in the domain of hyposensitivity, compared with 33.3% of children with language delay. The auditory abnormalities in autism primarily c...

Ying-Hua Tan; Chun-Yan Xi; Shu-Ping Jiang; Bing-Xin Shi; Li-Bo Wang; Lin Wang

2012-01-01

315

Vascular abnormalities in diabetes and their treatment  

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The capillary abnormalities of diabetes include both dilatation and occlusion. Occlusion is the more important, as vision-threatening lesions are secondary to capillary occlusion. Capillary occlusion could result from abnormalities of blood coagulation and from endothelial cell disease. Evidence is presented of the predominant role of endothelial disease--abnormal coagulation is probably secondary. While there is no known treatment which can reliably prevent or reverse capillary changes, appr...

Porta, Massimo

1980-01-01

316

Endothelin-3 like immunoreactivity in plasma of patients with cirrhosis of the liver  

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A highly specific and sensitive radioimmunoassay (RIA) has been established for determination of endothelin-3 like immunoreactivity in human plasma to investigate its possible role in hemodynamic alterations due to liver disease. Crossreactivity with other endothelin isoforms was always below 4 %, the lower detection limit following extraction on Sep-Pak C18 cartridges was 0.5 pg/ml. The concentration of endothelin-3 (mean ± SEM) was 4.16 ± 0.56 pg/ml (n = 13) in plasma of patients with cir...

1992-01-01

317

Presence of dynorphin-like immunoreactivity but not opiate binding in Walker-256 tumors  

International Nuclear Information System (INIS)

Walker-256 tumor tissue was removed from rats on day 8 of tumor growth. An acidified methanol extract of the tumor tissue was assayed for immunoreactive (ir) dynorphin-A 1-17 (DYN-17) and ir-dynorphin-A (DYN-8). Levels of ir-DYN-17 and ir-DYN-8 were nearly 4- and 8-fold higher, respectively, in tumors versus normal muscle. However, tumor homogenates did not exhibit specific "3H-naloxone binding. These results indicate that although the Walker-256 carcinosarcoma may produce opioids, it is unlikely that these ectopic substances have direct opioid actions on the tumor itself. 34 references, 1 figure

1985-07-15

318

Topography and time course of changes in spinal neuropeptide Y immunoreactivity after spared nerve injury  

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We used a new computer-assisted method to precisely localize and efficiently quantify increases in NPY immunoreactivity (NPY-ir) along the mediolateral axis of the L4 dorsal horn following transection of either the tibial and common peroneal nerves (thus sparing the sural branch, spared nerve injury, SNI), the tibial nerve, or the common peroneal and sural nerves. Two weeks after SNI, NPY-ir increased within the tibial and peroneal innervation territories; however, NPY-ir in the central-later...

Intondi, A. B.; Zadina, J. E.; Zhang, X.; Taylor, B. K.

2010-01-01

319

Marked digoxin-like immunoreactive factor interference with an enzyme immunoassay.  

Science.gov (United States)

A case in which digoxin-like immunoreactive factors (DLIF) interfered with an enzyme immunoassay in a patient with renal insufficiency is reported. A 79-year-old woman was found to have a serum digoxin concentration (SDC) determined by enzyme immunoassay of 5.0 ng/ml. Although all subsequent SDC determined by the enzyme immunoassay system were elevated, identical samples run on a fluorescence polarization immunoassay revealed SDC within the therapeutic range. Marked DLIF-related assay interference has been reported to occur with some digoxin assays; however, the enzyme immunoassay methods have never been reported to cross-react to the magnitude seen in this case. PMID:3063480

Karboski, J A; Godley, P J; Frohna, P A; Horton, M W; Reitmeyer, W J

1988-09-01

320

Influence of digoxin-like immunoreactive factor on late complications in patients with diabetes mellitus  

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The aim of this study was to compare the intensity of typical late complications in diabetic patients (n = 65, 28 type I, 37 type II) who were not on glycoside drugs with low vs. high serum levels of digoxin-like immunoreactive factor (DLIF: group I, n = 42, DLIF < or = the detection limit of 0.2 ng ml-1; and group II, n = 23, mean +/- SEM: 1.17 +/- 0.31 [0.25-4.96] ng ml-1). For detection of nephropathy, urinary albumin excretion (24 h) and creatinine clearance tests were used. For coronary ...

1994-01-01

 
 
 
 
321

Endogenous digoxin-like immunoreactivity in follicular fluid and in vitro fertilization.  

Science.gov (United States)

Plasma digoxin-like immunoreactive factor (DLIF) has been detected in various pathophysiological conditions associated with volume expansion. In this study, using radioimmunoassay, we confirmed the existence of high levels of DLIF in the stimulated follicular fluid, a rapidly volume-expanding biological model. The concentration of the various fractions of DLIF in follicular fluid was 2-9 times higher than in plasma, suggesting local concentration or production. No difference in concentration was observed between follicles containing fertilized oocytes and follicles with unfertilized oocytes. The role of DLIF in follicular homeostasis remains to be further investigated. PMID:1663910

Jakobi, P; Krivoy, N; Eibschitz, I; Ziskind, G

1991-01-01

322

Rapid radioimmunoassay for human immunoreactive pancreatic phospholipase A2 and its normal values  

International Nuclear Information System (INIS)

A rapid radioimmunoassay was developed for measuring immunoreactive pancreatic phospholipase A2 (IR-P-PLA2) in human sera. All analytic procedures could be accomplished within 3 hours. The assay was shown to be reproducible, sensitive and specific. The IR-P-PLA2 values were 7.39 ± 2.86 ng/ml in 111 healthy subjects. In the patients with chronic renal failure, the IR-P-PLA2 contents were significantly higher and did not decrease by blood dialysis

1990-01-01

323

Transforming growth factor alpha-immunoreactivity in neural tissues of the rat stomach.  

Science.gov (United States)

We report TGF alpha immunoreactivity in neurons of the myenteric plexus and in nerve fibers in the muscle and submucosal layers of the rat stomach. Association of TGF alpha staining nerve fibers to vessels and smooth muscle cells gives morphological evidence that EGF/TGF alpha's actions to increase mucosal blood flow and gastric motility may be mediated by TGF alpha derived from neural structures. These data suggest that TGF alpha plays a role in the neural control of the gastric function. PMID:9226395

Hoffmann, P; Zeeh, J M; Lakshmanan, J; Liu, L; Cooray, D; Barajas, L; Eysselein, V E

1997-04-30

324

A new look at calretinin-immunoreactive amacrine cell types in the monkey retina  

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We have examined amacrine cells that are calretinin-immunoreactive (-IR) in the macaque monkey retina with the aim of classifying them into morphological and functional subtypes. There are calretinin-IR cells in the fovea and throughout the retina. Their highest density is reached at 1.0 mm from the foveal pit (10,500 cells/mm2) and falls to 2,600/mm2 by 10 mm of eccentricity. Nearest-neighbor statistics for the calretinin-IR cell body distribution indicate a nonregular pattern, with a regula...

Kolb, Helga; Zhang, Li; Dekorver, Laura; Cuenca Navarro, Nicola?s

2002-01-01

325

Modulation of glial fibrillary acidic protein immunoreactivity during Sparus aurata L. development. A preliminary report  

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[EN] A band of approximately 50-5 1 kDa, corresponding to glial fibrillary acidic protein (GFAP), was detected by immunoblot techniques in the brain tissue of Sparus aurata L. The levels of GFAP immunoreactivity were determined at three different stages of development: 6 month old, 18 month old and adults reared in an aquaculture facility for 3-4 years. Our results indicated a decrease of GFAP levels during S. aurata development from six month old to adult individuals. An explanation for this...

Busquets, Xavier; Morales-nin, Beatriz; Ventayol, Pedro

1996-01-01

326

Effect of early oral calcium supplementation on serum calcium and immunoreactive calcitonin concentration in preterm infants.  

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Oral calcium supplements (80 mg/kg per 24 h) were given to 23 preterm infants, and the course of serum calcium, magnesium, immunoreactive calcitonin, and gastrin was compared with a control group of 23 matched infants. In the supplemented group, serum calcium concentrations remained at the baseline level (2.31 mmol/l +/- 0.18 SD) while a fall (from 2.27 +/- 0.18 to 1.91 +/- 0.24 mmol/l) was observed at 12-16 hours of age in the control group, with 4 values < 1.75 mmol/l. There was no change i...

Sann, L.; David, L.; Chayvialle, J. A.; Lasne, Y.; Bethenod, M.

1980-01-01

327

Ventricular somatostatin-like immunoreactivity in patients with basal ganglia disease.  

Science.gov (United States)

The concentrations of somatostatin-like immunoreactivity (SLI) in lateral ventricular fluid of patients with extrapyramidal motor disease were determined by specific radio-immunoassay. Mean SLI levels were significantly lower in patients with Parkinson's disease (mean +/- SEM); 42.9 +/- 2.9 fmol/ml) and in patients with dystonic syndromes (39.4 +/- 3.2) than in patients with benign essential tremor (65.3 +/- 9.7). The lowest levels were found in patients with athetosis (34.7 +/- 5.4). In parkinsonian patients somatostatin levels correlated with the degree of akinesia, rigidity and autonomic disturbances. PMID:2864402

Cramer, H; Wolf, A; Rissler, K; Weigel, K; Ostertag, C

1985-01-01

328

Fetal facial abnormalities identified during obstetric sonography.  

Science.gov (United States)

The orbits and, if readily accessible, the facial profile were visualized as part of a brief fetal anatomy survey during approximately 7100 low-risk and high-risk obstetric sonographic examinations. This examination identified 17 facial abnormalities in 11 fetuses. There were seven abnormalities of the eyes, including three instances of absence of both eyes, two of hypertelorism, one of proptosis, and one of relative prominence. There were seven abnormalities of the nose, including three instances of marked flattening, one of absence, one of a proboscis, one of a single nostril, and one of a sunken appearance. There were two abnormally small chins. There was one marked cleft that involved the nose, lip, and palate. At least two fetuses with abnormal faces were missed entirely and coexistent facial abnormalities were missed in another three fetuses. Of the 11 fetuses with facial abnormalities identified, eight had other structural abnormalities as well, and the other three had either polyhydramnios or a history of teratogen exposure. A brief facial examination done as part of the fetal anatomy survey helps to identify abnormalities in high-risk fetuses but is considerably less productive in low-risk fetuses. PMID:3543386

Hegge, F N; Prescott, G H; Watson, P T

1986-12-01

329

Radiologic atlas of pulmonary abnormalities in children  

International Nuclear Information System (INIS)

This book is an atlas about thoracic abnormalities in infants and children. The authors include computed tomographic, digital subtraction angiographic, ultrasonographic, and a few magnetic resonance (MR) images. They recognize and discuss how changes in the medical treatment of premature infants and the management of infection and pediatric tumors have altered some of the appearances and considerations in these diseases. Oriented toward all aspects of pulmonary abnormalities, the book starts with radiographic techniques and then discusses the normal chest, the newborn, infections, tumors, and pulmonary vascular diseases. There is comprehensive treatment of mediastinal abnormalities and a discussion of airway abnormalities

1988-01-01

330

Sensorial abnormalities: Smell and taste  

Directory of Open Access Journals (Sweden)

Full Text Available Introduction: Taste and smell abnormalities have proven to be an extremely more complex subject than previously regarded. Wide-ranging nosologic entities arise along with smell and taste alterations, and they can be congenital or acquired. Objective: Analyze the main features of smell and taste dysfunctions. Method: Automated databases were used to collect data, by searching keywords like 'alteration', 'smell', and 'taste'. A non-systematic search was also made in scientific printings and medical books. Literature Review: Smell and taste dysfunctions have a vast etiology, the most significant of which are obstructive nasal and sinusal disease, infections of the upper respiratory tract, cranioencephalic trauma, aging, exposure to toxics and some drugs, nasal or intracranial neoplasias, psychiatric and neurological pathologies, iatrogenic disease, idiopathic and congenital causes. A detailed anamnesis, a careful physical examination and supplementary evaluations are important for the diagnosis of these alterations. Conclusion: As a rule, smell and taste dysfunctions occur in a combined way. The early discovery of such dysfunctions can lead to a more efficient treatment, making the progress of diseases causing them retard and the symptoms less severe. In many cases, treating these alterations is not easy and there needs to be a multidisciplinary cooperation among the otorhinolaryngologist, endocrinologist, neurologist, psychiatrist, among others.

Palheta Neto, Francisco Xavier

2011-07-01

331

Characterization of immunoreactive trypsin as a means of differentiating graft pancreatitis and allograft rejection after porcine pancreatic transplantation.  

Science.gov (United States)

Graft pancreatitis and allograft rejection were both accompanied by increased serum levels of immunoreactive anionic trypsin (irAT) in a porcine pancreatic allograft transplantation model. Characterization of this immunoreactivity by gel filtration revealed different elution profiles in these conditions that can be helpful in the differentiation between them. During graft pancreatitis, a major part of the immunoreactivity was found within the high-molecular-weight fraction corresponding to the formation of complexes between trypsin and protease inhibitors. During allograft rejection, virtually all serum irAT increase could be attributed to the release of anionic trypsinogen without any evidence of activation. Since this transplantation model includes urinary diversion of the exocrine secretions, irAT and immunoreactive cationic trypsin (irCT) can also be measured in the urine. Characterization of this immunoreactivity showed that most of both irAT and irCT was found as active trypsin but a minor part was probably complexed with some protease inhibitor (possibly pancreatic secretory trypsin inhibitor [PSTI]). PMID:1733080

Källén, R; Borgström, A

1992-01-01

332

Blood plasma magnesium, potassium, glucose, and immunoreactive insulin changes in cows moved abruptly from barn feeding to early spring pasture  

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Cations and immunoreactive insulin in plasma were measured in 35 lactating cows moved abruptly to early spring pasture. After change of cows from grass-clover hay to fescue-bluegrass pasture containing 22 to 31 g potassium/kg dry matter, immunoreactive insulin of 5 Holstein cows increased 30% in 5 days and averaged 45% above prepasture concentrations for 40 days. Magnesium averaged 44% below prepasture content of plasma during this period and was correlated negatively with potassium -.17 and immunoreactive insulin -.37. Thirty Hereford cows were changed from corn silage and grass-clover hay to wheat-rye pasture containing 3.06% potassium in the dry matter. Each day on pasture, 10 cows each were fed 2.3 kg cornmeal, 10 were given 30 g magnesium oxide by capsule, and 10 were given no supplement. After unsupplemented cows were moved to pasture, immunoreactive insulin rose 51% in 8 days and plasma magnesium fell 24%. Both supplements reduced immunoreactive insulin, but magnesium was maintained higher by magnesium oxide than by cornmeal. Injection of two Holstein cows with insulin (2 IU/kg body weight) reduced plasma concentrations of both potassium and mgnesium 20% below that of two cows injected with only physiological saline. Whether elevated plasma insulin may accelerate development of hypomagnesemia in cattle on spring pasture with relatively high potassium content has not been established.

Miller, J.K. (Comparative Animal Research Lab., Oak Ridge, TN); Madsen, F.C.; Lentz, D.E.; Wong, W.O.; Ramsey, N.; Tysinger, C.E.; Hansard, S.L.

1980-07-01

333

Distribution of glucagon-like peptide (GLP)-2-immunoreactive cells in the chicken small intestine: antigen retrieval immunohistochemistry.  

Science.gov (United States)

An antigen retrieval method for immunohistochemical staining of glucagon-like peptide (GLP)-2-immunoreactive cells was investigated in the chicken small intestine. GLP-2-immunoreactive cells were observed as open-typed endocrine cells in the villous epithelium and crypts on both antigen retrieval agent-treated and untreated preparations. No obvious differences were detected in morphological features of GLP-2-immunoreactive cells between treated and untreated preparations. The frequencies of occurrence of GLP-2-immunoreactive cells, however, were significantly different in treated and untreated preparations: in the proximal and distal regions of jejunum and ileum obtained from untreated preparations, the frequencies of occurrence were 0.5 ± 0.2, 0.7 ± 0.1, 0.9 ± 0.2 and 1.5 ± 0.3, respectively (cell numbers per mucosal area: cells/mm(2), mean ± SD), whereas those from treated sections were 14.7 ± 2.3, 19.8 ± 2.3, 23.5 ± 4.7 and 34.6 ± 4.9 cells/mm(2), respectively. These data indicate that this antigen retrieval method is able to make immunoreactive GLP-2 available for detection and that GLP-2 may act as one of the common hormones secreted by L cells in the chicken small intestine. PMID:24334814

Monir, Mohammad M; Hiramatsu, Kohzy; Nishimura, Kei; Takemoto, Chihiro; Watanabe, Takafumi

2014-04-01

334

Immunohistochemical detection of ganglia in the rat stomach serosa, containing neurons immunoreactive for gastrin-releasing peptide and vasoactive intestinal peptide  

DEFF Research Database (Denmark)

Ganglia, not previously described, were identified in the rat stomach serosa along the minor curvature. The ganglia consisted of varying number of cell bodies lying in clusters along or within nerve bundles. The ganglia were shown to contain GRP and VIP immunoreactive nerve fibers and cell bodies and also some NPY immunoreactive fibers, whereas they were devoid of somatostatin immunoreactivity. Nerve ligation experiments indicated that the ganglia are intrinsic to the stomach.

Poulsen, Steen Seier; Holst, J J

1987-01-01

335

Does the in vitro testing of the immunoreactivity of an antibody reflect its in vivo behavior  

International Nuclear Information System (INIS)

It has been assumed that radioimmunoassays (RIA) can be used as an in vitro test to predict the integrity of radiolabeled antibody molecules. the effects of radioiodination of antibody molecules on their in vitro immunoreactivity and in vivo biodistribution in normal and tumor bearing animals. Several immunoglobulins (IgG) were radiolabeled with I125 in the presence of chloramine T (CT), and the molar iodine to antibody ratios were varied from 1:1 to 35:1. One of the IgG was also internally radiolabeled using Se75-selenomethionine. While no differences in immunoreactivity were observed in vitro for iodine: antibody ratios of up to 25:1, the in vivo biodistribution of the antibodies was dependent on the number of iodine atoms per molecule. Therefore, in vitro quality testing of a radiolabeled antibody does not always predict in vivo behavior. The results indicate that there is a need for identifying the variables involved in current radioimmunoassays and for developing other in vitro assay systems that can predict in vivo behavior

1988-01-01

336

Elevated serum levels of immunoreactive anionic trypsin (but not cationic trypsin) signals pancreatic disease.  

Science.gov (United States)

Pancreatic juice from most studied species contains two major forms of trypsin, one with anionic electrophoretic mobility and one with cationic mobility. They are referred to as anionic and cationic trypsin(ogen). The purpose of this study was to measure immunoreactive anionic trypsin (irAT) and immunoreactive cationic trypsin (irCT) in sera from patients with pancreatic cancer (n = 39) and chronic pancreatitis (n = 32) using two specific ELISA methods. Sera from 72 healthy persons were used as controls. Patients with pancreatic cancer showed significantly elevated serum levels of irAT median level 39 vs 20.5 micrograms/L in the control group (p irAT and irCT in serum was significantly increased (p irAT and irCT levels, but no significant differences compared to the control group. The ratio between irAT and irCT was, however, significantly increased also in this group of patients. The results suggest a nonparallel secretion of anionic and cationic trypsinogen in pancreatic disease. This is a pattern that has been observed in experimental forms of chronic "hyperCCKemia." PMID:8708393

Borgström, A; Andrén-Sandberg, A

1995-12-01

337

Depletion of somatostatin-like immunoreactivity in the rat central nervous system by cysteamine  

International Nuclear Information System (INIS)

Selective neurotoxins have been of value in providing a means for specifically interfering with the actions of endogenous neurotransmitter candidates. Others have shown cysteamine (CSH) to deplete the gastrointestinal tract and hypothalamus of rats of immunoreactive somatostatin, suggesting a toxic action of that compound directed against somatostatin-containing cells. The present study further defines the actions of cysteamine on somatostatin in the central nervous system. (CNS). Cysteamine hydrochloride administered subcutaneously results in a depletion of somatostatin-like immunoreactivity (SLI) in the retina, brain, and cervical spinal cord of rats. The effect is demonstrable at doses of 30 mg/kg of body weight and above, occurs within 2 to 4 hr of a single injection of the drug, and is largely reversible within 1 week. The mean depletion of SLI observed within the CNS varies from 38% in cerebral cortex to 65% in cervical spinal cord 24 hr following administration of CSH, 300 mg/kg of body weight, s.c. By gel permeation chromatography, all molecular weight forms of SLI are affected, with the largest reductions in those forms that co-chromatograph with synthetic somatostatin-14 and somatostatin-28. These results indicate that CSH has a generalized, rapid, and largely reversible effect in depleting SLI from the rat CNS

1982-01-01

338

Colocalization of immunoreactive oxytocin, vasopressin and interleukin-1 in human thymic epithelial neuroendocrine cells.  

Science.gov (United States)

Monoclonal antibodies to oxytocin (OT) and vasopressin (VP) revealed some positively staining stromal cells in the subcapsular cortex and in the medulla of the human thymus. We further demonstrated that these cells are a subset of epithelial endocrine cells and also contain immunoreactive interleukin-1 together with the neuropeptides. In addition, the thymic cells stained by monoclonal antibodies directed to the cyclic part of oxytocin or vasopressin also contained some immunoreactive neurophysins. These data support the concept of intrathymic synthesis of neurohypophyseal-like peptides fitting the hypothalamic model. However, we observed that, contrary to the situation in the brain, OT- and VP-like peptides colocalized in the same thymic cells. Furthermore, one monoclonal antibody, specific for the tail part of oxytocin, did not label thymic cells. Therefore, thymic neuropeptide(s) could be related to, but distinct from, authentic OT and VP. These observations suggest some molecular differences between hypothalamic and thymic oxytocin biosynthetic pathways which need to be further investigated. PMID:1712650

Robert, F; Geenen, V; Schoenen, J; Burgeon, E; De Groote, D; Defresne, M P; Legros, J J; Franchimont, P

1991-03-01

339

Egg-laying-hormone immunoreactivity in the neural ganglia and ovary of Haliotis asinina Linnaeus.  

Science.gov (United States)

Immunoreactivity against the abalone egg-laying hormone (aELH) was detected in the fine granules of type 1 and 2 neurosecretory (NS) cells, neurites in the neuropil, and blood sinuses in the connective tissue sheath of the cerebral, pleuropedal, and visceral ganglia of the tropical abalone, Haliotis asinina Linnaeus. The number of positive NS cells, and the intensity of staining in the ganglia, varied and might be related to the stage of ovarian cycle. At any stage, positive cells were most numerous in the pleuropedal, and least numerous in the visceral ganglion. In addition, several cells of the statocyst and associated nerves also exhibited the immunoreactivity. In the ovary, the most intense reactivity was detected in the follicular and granular cells adjacent to mature oocytes, in the trabeculae and the ovarian capsule. The cytoplasm of mature oocytes was also moderately stained. The results indicate that the cerebral, pleuropedal, and visceral ganglia are the main sites of aELH-producing cells. The ovary may also produce aELH locally. PMID:15803317

Saitongdee, P; Apisawetakan, S; Anunruang, N; Poomthong, T; Hanna, P; Sobhon, P

2005-11-01

340

Initiation of chemical studies on the immunoreactive glycolipids of adult Ascaris suum.  

Science.gov (United States)

There is a general lack of basic information concerning one class of glycoconjugate, the glycolipids, from parasitic nematodes. As the prototype, the neutral glycolipid fraction derived from adult males of Ascaris suum was investigated as to its chromatographic, differential chemical staining, antigenic and chemical properties. The thin-layer chromatography-resolved neutral fraction glycolipids could be classified into components of fast and slow migrating band groups. Immunoreactivity was restricted to the latter as detected by IgG and IgM anti-neutral fraction glycolipid antibody levels in serial infection sera of mice. Similarities of chromatography, antigenicity and serological cross-reactivity have been extended to the neutral glycolipid fractions of other parasitic nematodes: Litomosoides carinii and Nippostrongylus brasiliensis. Chemical, differential chemical staining and enzymatic analyses identified the Ascaris suum antigenic, slow migrating band group of components as amphoteric glycosphingolipids, and not the originally hypothesized glycoglycerolipids or glycosylphosphatidylinositols, that contained typical neutral monosaccharide constituents and a zwitterionic phosphodiester linkage, most probably phosphocholine. Glycosphingolipid-immunoreactivity is eliminated on cleavage of the zwitterionic phosphodiester linkage by hydrofluoric acid treatment. PMID:7596644

Dennis, R D; Baumeister, S; Smuda, C; Lochnit, C; Waider, T; Geyer, E

1995-06-01

 
 
 
 
341

Stability and immunoreactivity of the monoclonal anticytokeratin antibody TS1 after different degrees of iodination  

Energy Technology Data Exchange (ETDEWEB)

The immunoreactivity, stability and in vivo kinetics of an anticytokeratin 8 monoclonal antibody, TS1, were investigated following different degrees of labeling with {sup 125}I (0.2, 1 and 2-3 {sup 125}I/TS1 MAb). By testing with ELISA, it was demonstrated that a high degree of iodination, i.e. > 2 {sup 125}I/TS1, caused a rapid decrease in immunoreactivity to almost zero within 10 days. Furthermore, a complete degradation to low molecular weight fragments and free iodine was seen, as shown by SDS PAGE and autoradiography. The differently labeled radionuclide conjugates were injected into nude mice inoculated with HeLa Hep2 cells and tumor doses (estimated by MIRD formalism), tumor:non-tumor dose ratios, % I.D./gram tissue, Gy/MBq and in vivo kinetics of the differently labeled MAbs were determined. Despite the in vitro instability of the highest iodinated radionuclide conjugate, it was possible to deliver high doses to the tumors if the conjugate was injected into the animal immediately after completion of the iodination procedure. Increases from 1.4 Gy to 15.2 Gy delivered tumor dose were obtained with a tenfold increase in the specific activity, without alterations in the tumor:non-tumor tissue dose ratios. There is room for significant improvements in efficacy at radioimmunotherapy, which can be gained by optimizing the degree of iodination. For therapeutical applications a high degree of iodination may be an advantage. (orig.)

Johansson, A.; Sandstroem, P.; Ullen, A.; Stigbrand, T. [Umeaa Univ. (Sweden). Dept. of Immunology; Erlandsson, A.; Sundstroem, B. [Vaermland University Coll. of Health and Caring Sciences, Karlstad (Sweden). Dept. of Biomedical Laboratory Science; Riklund Aahlstroem, K.; Hietala, S.O. [Umeaa Univ. (Sweden). Dept. of Diagnostic Radiology; Johansson, L. [Umeaa Univ. (Sweden). Dept. of Radiation Physics

1999-11-01

342

Stability and immunoreactivity of the monoclonal anticytokeratin antibody TS1 after different degrees of iodination  

International Nuclear Information System (INIS)

The immunoreactivity, stability and in vivo kinetics of an anticytokeratin 8 monoclonal antibody, TS1, were investigated following different degrees of labeling with 125I (0.2, 1 and 2-3 125I/TS1 MAb). By testing with ELISA, it was demonstrated that a high degree of iodination, i.e. > 2 125I/TS1, caused a rapid decrease in immunoreactivity to almost zero within 10 days. Furthermore, a complete degradation to low molecular weight fragments and free iodine was seen, as shown by SDS PAGE and autoradiography. The differently labeled radionuclide conjugates were injected into nude mice inoculated with HeLa Hep2 cells and tumor doses (estimated by MIRD formalism), tumor:non-tumor dose ratios, % I.D./gram tissue, Gy/MBq and in vivo kinetics of the differently labeled MAbs were determined. Despite the in vitro instability of the highest iodinated radionuclide conjugate, it was possible to deliver high doses to the tumors if the conjugate was injected into the animal immediately after completion of the iodination procedure. Increases from 1.4 Gy to 15.2 Gy delivered tumor dose were obtained with a tenfold increase in the specific activity, without alterations in the tumor:non-tumor tissue dose ratios. There is room for significant improvements in efficacy at radioimmunotherapy, which can be gained by optimizing the degree of iodination. For therapeutical applications a high degree of iodination may be an advantage. (orig.)

1999-01-01

343

Purification and characterization of endogenous digoxin-like immunoreactive factors in chicken blood.  

Science.gov (United States)

Studies have been performed to determine whether an endogenous material capable of binding to digoxin antibodies is present in the chicken plasma. In the blood of 12 chickens without feed control, endogenous digoxin-like immunoreactive factors (DLIF) binding of digoxin antibodies in enzyme immunoassays amounted to 866 / 302 pg digoxin equivalents/mL of plasma (mean +/- SEM). Immunoreactivity of DLIF increased to 1848***331 pg/mL with a double value of control after boiling and acid pretreating the plasma. The major purification steps employed in this report were gel filtration column chromatography, high performance liquid chromatography (HPLC) and isoelectric focusing (IEF). Using HPLC for the separation, at least 10 chicken DLIFs with different molecular weight (MW) have been found. The MW of the smallest is 300 daltons (Da) while the largest is 100 kDa. The value of the isoelectric point of the most abundant type of DLIF from untreated chicken plasma is 6.3 as determined by IEF. The partially purified DLIF inhibits Na+, K(+)-ATPase from a porcine cerebral cortex as well as three human red blood cell membrane preparations in a dose-response fashion. PMID:8913327

Wei, J S; Cheng, H C; Tsai, K J; Liu, D H; Lee, H H; Chiu, D T; Liu, T Z

1996-01-01

344

Melatonin immunoreactivity in the photosynthetic prokaryote Rhodospirillum rubrum: implications for an ancient antioxidant system.  

Science.gov (United States)

Rhodospirillum rubrum is a spiral anoxygenic photosynthetic bacterium that can exist under either aerobic or anaerobic conditions. The organism thrives in the presence of light or complete darkness and represents one of the oldest species of living organisms, possibly 2-3.5 billion years old. The success of this prokaryotic species may be attributed to the evolution of certain indole compounds that offer protection against life-threatening oxygen radicals produced by an evolutionary harsh environment. Melatonin, N-acetyl-5-methoxytryptamine, is an indolic highly conserved molecule that exists in protists, plants, and animals. This study was undertaken to determine the presence of an immunoreactive melatonin in the kingdom Monera and particularly in the photosynthetic bacterium, R. rubrum, under conditions of prolonged darkness or prolonged light. Immunoreactive melatonin was measured during both the extended day and extended night. Significantly more melatonin was observed during the scotophase than the photophase. This study marks the first demonstration of melatonin in a bacterium. The high level of melatonin observed in bacteria may provide on-site protection of bacterial DNA against free radical attack. PMID:8867786

Manchester, L C; Poeggeler, B; Alvares, F L; Ogden, G B; Reiter, R J

1995-01-01

345

p53 immunoreactivity correlates with Ki-67 and bcl-2 expression in renal cell carcinoma.  

Science.gov (United States)

This study assessed the relation of proliferation, inhibition of apoptosis, and the p53 tumor suppressor protein expression in clear renal cell carcinoma (RCC). Archival pathological specimens from 43 patients treated for RCC were obtained. Median follow-up for the patients was 52 months (range 2.5 months to 178 months). Immunostaining of paraffin tissue sections was carried out for four different markers: a) Ki-67, a marker for cellular proliferation; b) p53/DO7, c) p53/pAb240, antibodies for the p53 protein; and d) bcl-2, a marker for inhibition of apoptosis (programmed cell death). One thousand cells were counted per slide at 400x magnification. Staining of >/=10% of cells was considered positive and <10% negative. Fisher exact contingency tables were used for correlation between markers, tumor grade and stage. A significant correlation was found between Ki-67 and p53 immunoreactivity samples, P=0.0001. Interestingly, a significant association was found if Ki-67 and bcl-2 scores were combined and correlated with p53, P=0.009. Results showed no correlation between any of the immunohistochemical markers and grade or stage. In addition, Kaplan-Meier survival curves demonstrated no significant difference between patients' tumors that was scored immunoreactive negative vs. positive for Ki-67, p53, or bcl-2. This study indicates that p53 expression correlates with proliferation, and inhibition of programmed cell death in RCC. PMID:11166623

Olumi, A F.; Weidner, N; Presti, J C.

2001-03-01

346

Neuropeptide Y-immunoreactive neurons in the cerebral cortex of humans and other haplorrhine primates.  

Science.gov (United States)

We examined the distribution of neurons immunoreactive for neuropeptide Y (NPY) in the posterior part of the superior temporal cortex (Brodmann's area 22 or area Tpt) of humans and nonhuman haplorrhine primates. NPY has been implicated in learning and memory and the density of NPY-expressing cortical neurons and axons is reduced in depression, bipolar disorder, schizophrenia, and Alzheimer's disease. Due to the role that NPY plays in both cognition and neurodegenerative diseases, we tested the hypothesis that the density of cortical and interstitial neurons expressing NPY was increased in humans relative to other primate species. The study sample included great apes (chimpanzee and gorilla), Old World monkeys (pigtailed macaque, moor macaque, and baboon) and New World monkeys (squirrel monkey and capuchin). Stereologic methods were used to estimate the density of NPY-immunoreactive (-ir) neurons in layers I-VI of area Tpt and the subjacent white matter. Adjacent Nissl-stained sections were used to calculate local densities of all neurons. The ratio of NPY-ir neurons to total neurons within area Tpt and the total density of NPY-ir neurons within the white matter were compared among species. Overall, NPY-ir neurons represented only an average of 0.006% of the total neuron population. While there were significant differences among species, phylogenetic trends in NPY-ir neuron distributions were not observed and humans did not differ from other primates. However, variation among species warrants further investigation into the distribution of this neuromodulator system. PMID:23042407

Raghanti, Mary Ann; Conley, Tiffini; Sudduth, Jessica; Erwin, Joseph M; Stimpson, Cheryl D; Hof, Patrick R; Sherwood, Chet C

2013-05-01

347

Estrophilin immunoreactivity versus estrogen receptor binding activity in meningiomas: evidence for multiple estrogen binding sites  

International Nuclear Information System (INIS)

The existence of estrogen receptors in human meningiomas has long been a controversial issue. This may be explained, in part, by apparent heterogeneity of estrogen binding sites in meningioma tissue. In this study, estrogen receptors were determined in 58 meningiomas with an enzyme immunoassay using monoclonal antibodies against human estrogen receptor protein (estrophilin) and with a sensitive radioligand binding assay using 125I-labeled estradiol (125I-estradiol) as radioligand. Low levels of estrophilin immunoreactivity were found in tumors from 62% of patients, whereas radioligand binding activity was demonstrated in about 46% of the meningiomas examined. In eight (14%) tissue samples multiple binding sites for estradiol were observed. The immunoreactive binding sites correspond to the classical, high affinity estrogen receptors: the Kd for 125I-estradiol binding to the receptor was approximately 0.2 nM and the binding was specific for estrogens. The second, low affinity class of binding sites considerably influenced measurement of the classical receptor even at low ligand concentrations. The epidemiological and clinical data from patients with meningiomas, and the existence of specific estrogen receptors confirmed by immunochemical detection, may be important factors in a theory of oncogenesis

1987-01-01

348

Increase in GAP-43 and GFAP immunoreactivity in the rat hippocampus subsequent to perforant path kindling  

DEFF Research Database (Denmark)

Kindling is an animal model of epilepsy which is accompanied by morphological and biochemical changes in the brain, including sprouting of fibers and increased transmitter release. Here we have examined the immunocytochemical expression of 1) GAP-43, a growth-associated protein, which is a neuron-specific PKC substrate, particularly expressed in development and regeneration and 2) glial fibrillary acidic protein (GFAP), part of the astrocytic cytoskeleton, after perforant path kindling. Subsequent to kindling, GAP-43 immunoreactivity was increased in CA1 stratum lacunosum-moleculare and the inner and outer molecular layer of the fascia dentata. Other hippocampal subregions showed a lower increase. GFAP immunoreactivity was increased in the entire hippocampus, but especially in stratum lacunosum-moleculare of the CA1 and the hilus of fascia dentata. The difference between the number of GFAP-positive profiles in the hippocampus of control rats and in fully kindled rats was found to be non-significant. We interpret these findings as being related to both plastic neuronal changes and possible neuronal degeneration.

Dalby, Nils Ole; Rondouin, G

1995-01-01

349

Irisin-immunoreactivity in neural and non-neural cells of the rodent.  

Science.gov (United States)

Irisin is a recently identified myokine secreted from the muscle in response to exercise. In the rats and mice, immunohistochemical studies with an antiserum against irisin peptide fragment (42-112), revealed that irisin-immunoreactivity (irIRN) was detected in three types of cells; namely, skeletal muscle cells, cardiomyocytes, and Purkinje cells of the cerebellum. Tissue sections processed with irisin antiserum pre-absorbed with the irisin peptide (42-112) (1 ?g/ml) showed no immunoreactivity. Cerebellar Purkinje cells were also immunolabeled with an antiserum against fibronectin type II domain containing 5 (FNDC5), the precursor protein of irisin. Double-labeling of cerebellar sections with irisin antiserum and glutamate decarboxylase (GAD) antibody showed that nearly all irIRN Purkinje cells were GAD-positive. Injection of the fluorescence tracer Fluorogold into the vestibular nucleus of the rat medulla retrogradely labeled a population of Purkinje cells, some of which were also irIRN. Our results provide the first evidence of expression of irIRN in the rodent skeletal and cardiac muscle, and in the brain where it is present in GAD-positive Purkinje cells of the cerebellum. Our findings together with reports by others led us to hypothesize a novel neural pathway, which originates from cerebellum Purkinje cells, via several intermediary synapses in the medulla and spinal cord, and regulates adipocyte metabolism. PMID:23470775

Dun, S L; Lyu, R-M; Chen, Y-H; Chang, J-K; Luo, J J; Dun, N J

2013-06-14

350

Advances in understanding the molecular basis of frontotemporal dementia  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Frontotemporal dementia (FTD) is a clinical syndrome with a heterogeneous molecular basis. Until recently, the underlying cause was known in only a minority of cases that were associated with abnormalities of the tau protein or gene. In 2006, however, mutations in the progranulin gene were discovered as another important cause of familial FTD. That same year, TAR DNA-binding protein 43 (TDP-43) was identified as the pathological protein in the most common subtypes of FTD and amyotrophic later...

2012-01-01

351

Recent advances in the molecular basis of frontotemporal dementia  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Frontotemporal dementia (FTD) is a clinical syndrome with heterogeneous molecular basis. Until recently, our knowledge was limited to a minority of cases associated with abnormalities of the tau protein or gene (MAPT). However, in 2006, mutations in progranulin (GRN) were discovered as another important cause of familial FTD. That same year, TAR DNA binding protein 43 (TDP-43) was identified as the pathological protein in the most common subtypes of FTD and ALS. Since then, significant effort...

2012-01-01

352

Abnormal Mitochondrial Dynamics and Neurodegenerative Diseases  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Abstract Mitochondrial dysfunction is a prominent feature of various neurodegenerative diseases. A deeper understanding of the remarkably dynamic nature of mitochondria, characterized by a delicate balance of fission and fusion, has helped to fertilize a recent wave of new studies demonstrating abnormal mitochondrial dynamics in neurodegenerative diseases. This review highlights mitochondrial dysfunction and abnormal mitochondrial dynamics in Alzheimer disease, Parkinson disease, a...

2009-01-01

353

Abnormal Mitochondrial Dynamics and Neurodegenerative Diseases  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Mitochondrial dysfunction is a prominent feature of various neurodegenerative diseases. A deeper understanding of the remarkably dynamic nature of mitochondria, characterized by a delicate balance of fission and fusion, has helped to fertilize a recent wave of new studies demonstrating abnormal mitochondrial dynamics in neurodegenerative diseases. This review highlights mitochondrial dysfunction and abnormal mitochondrial dynamics in Alzheimer disease, Parkinson disease, amyotrophic lateral s...

Su, Bo; Wang, Xinglong; Zheng, Ling; Perry, George; Smith, Mark A.; Zhu, Xiongwei

2010-01-01

354

A new look at abnormal uterine bleeding.  

Science.gov (United States)

New universal terminology, classifications, and definitions recommended by the International Federation of Gynecology and Obstetrics and supported by the American College of Obstetricians and Gynecologists to describe abnormal uterine bleeding abnormalities in reproductive women are presented. Identification and management of anovulatory and ovulatory uterine bleeding are explored. PMID:24177024

Twiss, Janice J

2013-12-10

355

Losses of immunoreactive parvalbumin amacrine and immunoreactive alphaprotein kinase C bipolar cells caused by methylmercury chloride intoxication in the retina of the tropical fish Hoplias malabaricus  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english To quantify the effects of methylmercury (MeHg) on amacrine and on ON-bipolar cells in the retina, experiments were performed in MeHg-exposed groups of adult trahiras (Hoplias malabaricus) at two dose levels (2 and 6 µg/g, ip). The retinas of test and control groups were processed by mouse anti-parv [...] albumin and rabbit anti-alphaprotein kinase C (alphaPKC) immunocytochemistry. Morphology and soma location in the inner nuclear layer were used to identify immunoreactive parvalbumin (PV-IR) and alphaPKC (alphaPKC-IR) in wholemount preparations. Cell density, topography and isodensity maps were estimated using confocal images. PV-IR was detected in amacrine cells in the inner nuclear layer and in displaced amacrine cells from the ganglion cell layer, and alphaPKC-IR was detected in ON-bipolar cells. The MeHg-treated group (6 µg/g) showed significant reduction of the ON-bipolar alphaPKC-IR cell density (mean density = 1306 ± 393 cells/mm²) compared to control (1886 ± 892 cells/mm²; P

Bonci, D.M.O.; Lima, S.M.A. de; Grötzner, S.R.; Oliveira Ribeiro, C.A.; Hamassaki, D.E.; Ventura, D.F..

356

Abnormality diagnostic system for reactor recycling pump  

International Nuclear Information System (INIS)

The system of the present invention diagnosing the abnormality of a recycling pump in a BWR type reactor objectively and appropriately. That is, in a system for estimating a position where the abnormality occurred by analyzing vibrations of the recycling pump, the abnormality position is estimated by a neural network having vibration intensities on every predetermined frequency regions as the input. In the abnormality diagnosing systems, it has been important how to determine a causal matrix but the causal matrix has been determined by expert's subjective judgment. Further, since the system was not adaptable, it was difficult for the addition of observation data to the data base. However, in the system of the present invention, since the position where the abnormality occurred is estimated by the neural network, the subjective judgment can be excluded. Further, it can also cope with the addition of the data to the data bases. (I.S.)

1989-11-13

357

Abnormal Event Detection Using Local Sparse Representation  

DEFF Research Database (Denmark)

We propose to detect abnormal events via a sparse subspace clustering algorithm. Unlike most existing approaches, which search for optimized normal bases and detect abnormality based on least square error or reconstruction error from the learned normal patterns, we propose an abnormality measurement based on the difference between the normal space and local space. Specifically, we provide a reasonable normal bases through repeated K spectral clustering. Then for each testing feature we first use temporal neighbors to form a local space. An abnormal event is found if any abnormal feature is found that satisfies: the distance between its local space and the normal space is large. We evaluate our method on two public benchmark datasets: UCSD and Subway Entrance datasets. The comparison to the state-of-the-art methods validate our method's effectiveness.

Ren, Huamin; Moeslund, Thomas B.

2014-01-01

358

Report to Congress on abnormal occurrences  

International Nuclear Information System (INIS)

Section 208 of the Energy Reorganization Act of 1974 identifies an abnormal occurrence as an unscheduled incident or event that the Nuclear Regulatory Commission determines to be significant from the standpoint of public health and safety and requires a quarterly report of such events to be made to Congress. This report covers the period January through March 1993. There is one abnormal occurrence at a nuclear power plant disposed in this report that involved a steam generator tube rupture at Palo Verde Unit 2, and none for fuel cycle facilities. Three abnormal occurrences involving medical misadminstrations (two therapeutic and one diagnostic) at NRC-licensed facilities are also discussed in this report. No abnormal occurrences were reported by NRC's Agreement States. The report also contains information updating previously reported abnormal occurrences

1993-01-01

359

Desensitization and capsaicin-induced release of substance P-like immunoreactivity from guinea-pig ureter in vitro.  

Science.gov (United States)

Substance P-like immunoreactivity was assayed in superfusates of guinea-pig ureters following stimulation of afferent fibres with capsaicin, potassium chloride and the calcium channel agonist Bay K 8644. Capsaicin-evoked release of substance P-like immunoreactivity was calcium-dependent but unaffected by cobalt. Under appropriate conditions release was dose-related (ED50 = 610 nM) and reproducible. Selective desensitization to capsaicin could be demonstrated following prolonged exposure to different doses of capsaicin. No desensitization to capsaicin was observed following afferent fibre stimulation with a combination of Bay K 8644 and K+, which released a similar amount of substance P-like immunoreactivity as a desensitizing capsaicin stimulus. These data suggest that depletion of releasable substance P-like reactivity is unlikely to account for selective desensitization of ureteric primary afferent fibres to capsaicin. PMID:2477771

Dray, A; Hankins, M W; Yeats, J C

1989-01-01

360

Nesfatin-1/nucleobindin-2 like immunoreactivity in the olfactory system, brain and pituitary of frog, Microhyla ornata.  

Science.gov (United States)

Nesfatin-1 is a recently discovered anorectic protein derived from the precursor nucleobindin-2 (NUCB2). While nesfatin-1 has been widely studied in mammals, and goldfish, there are no reports of nesfatin-1 in amphibians. Using immunohistochemistry and Western blot analysis, this study assessed the distribution of NUCB2/nesfatin-1 in the brain of frog Microhyla ornata. NUCB2/nesfatin-1 like immunoreactivity was found in the olfactory receptor neurons, Bowman's glands and in the olfactory epithelium of medial diverticulum. In the brain, immunoreactive perikarya were seen in the anterior preoptic area, magnocellular nucleus, suprachiasmatic nucleus, ventromedial thalamic nucleus, posterior thalamic nucleus, nucleus infundibularis ventralis and dorsalis, and the cerebellar nucleus. NUCB2/nesfatin-1like immunoreactivity was also detected in the pineal and pituitary glands of frog. The presence of NUCB2/nesfatin-1 in the key brain regions suggest possible roles for this protein in the regulation of physiological processes in frogs. PMID:24768694

Senejani, A G; Gaupale, Tekchand C; Unniappan, Suraj; Bhargava, Shobha

2014-06-01

 
 
 
 
361

Homogeneous MGMT Immunoreactivity Correlates with an Unmethylated MGMT Promoter Status in Brain Metastases of Various Solid Tumors  

Science.gov (United States)

The O6-methylguanine-methyltransferase (MGMT) promoter methylation status is a predictive parameter for the response of malignant gliomas to alkylating agents such as temozolomide. First clinical reports on treating brain metastases with temozolomide describe varying effects. This may be due to the fact that MGMT promoter methylation of brain metastases has not yet been explored in depth. Therefore, we assessed MGMT promoter methylation of various brain metastases including those derived from lung (n?=?91), breast (n?=?72) kidney (n?=?49) and from malignant melanomas (n?=?113) by methylation-specific polymerase chain reaction (MS-PCR) and MGMT immunoreactivity. Fifty-nine of 199 brain metastases (29.6%) revealed a methylated MGMT promoter. The methylation rate was the highest in brain metastases derived from lung carcinomas (46.5%) followed by those from breast carcinoma (28.8%), malignant melanoma (24.7%) and from renal carcinoma (20%). A significant correlation of homogeneous MGMT-immunoreactivity (>95% MGMT positive tumor cells) and an unmethylated MGMT promoter was found. Promoter methylation was detected in 26 of 61 (43%) tumors lacking MGMT immunoreactivity, in 17 of 63 (27%) metastases with heterogeneous MGMT expression, but only in 5 of 54 brain metastases (9%) showing a homogeneous MGMT immunoreactivity. Our results demonstrate that a significant number of brain metastases reveal a methylated MGMT-promoter. Based on an obvious correlation between homogeneous MGMT immunoreactivity and unmethylated MGMT promoter, we hypothesize that immunohistochemistry for MGMT may be a helpful diagnostic tool to identify those tumors that probably will not benefit from the use of alkylating agents. The discrepancy between promoter methylation and a lack of MGMT immunoreactivity argues for assessing MGMT promoter methylation both by immunohistochemical as well as by molecular approaches for diagnostic purposes.

Ingold, Barbara; Schraml, Peter

2009-01-01

362

Isolation of digoxin-like immunoreactive factors from mammalian adrenal cortex.  

Science.gov (United States)

Endogenous digoxin-like immunoreactive factors (DLIF) are present in serum and tissues of humans and animals. To date, a tissue source for these factors has not been rigorously defined nor have these factors been isolated to identifiable homogeneity. In this study, we define the distribution of DLIF in mammalian tissues, demonstrate the adrenal cortex to be the principal source of this factor in bovine, and isolate DLIF to chromatographic homogeneity using high performance liquid chromatography (HPLC). DLIF concentrations in tissue extracts from rats measured as follows: adrenal glands, 44.3; serum, 6.3; liver, 5.2; kidney, 1.2; heart, brain, or lungs, less than 1.4 ng of digoxin-equivalent per g of protein. Human tissues showed similar results. In dogs, the ratio of the DLIF concentration in lumbar vein serum to that in infrarenal inferior vena cava serum was 3.3 +/- 0.4 (mean +/- S.E., n = 4). Bovine adrenal cortex contained 7 times more DLIF per g of tissue than the adrenal medulla. 70 +/- 4% (n = 7) of the total bovine cortical DLIF activity (6,159 pg of digoxin-equivalent) applied to a reverse phase HPLC column eluted as one definitive fraction. 60% of the digoxin-like immunoreactivity extracted from bovine serum also co-eluted with DLIF from adrenal. None of the 14 steroid molecules or 7 cardiac glycoside congeners co-eluted with the major DLIF activity. Our data indicate that 947 pmol of DLIF is equivalent to 1 pmol of digoxin-equivalent immunoreactivity. Preliminary mass spectral analysis suggests that purified DLIF has a molecular mass of 780 daltons comprised of one 390-dalton aglycone component plus several sugar moieties. This study establishes a definitive link between DLIF in serum and the adrenal cortex as a source tissue. We also demonstrate a method for purifying DLIF to chromatographic homogeneity with an extraction capacity of 1.2 nmol of DLIF per g of adrenal cortex. PMID:1856201

Shaikh, I M; Lau, B W; Siegfried, B A; Valdes, R

1991-07-25

363

Pericellular innervation of neurons expressing abnormally hyperphosphorylated tau in the hippocampal formation of Alzheimer's disease patients  

Directory of Open Access Journals (Sweden)

Full Text Available Neurofibrillary tangles (NFT represent one of the main neuropathological features in the cerebral cortex associated with Alzheimer’s disease (AD. This neurofibrillary lesion involves the accumulation of abnormally hyperphosphorylated or abnormally phosphorylated microtubule-associated protein tau into paired helical filaments (PHF-tau within neurons. We have used immunocytochemical techniques and confocal microscopy reconstructions to examine the distribution of PHF-tau-immunoreactive (ir cells, and their perisomatic GABAergic and glutamatergic innervations in the hippocampal formation and adjacent cortex of AD patients. Furthermore, correlative light and electron microscopy was employed to examine these neurons and the perisomatic synapses. We observed two patterns of staining in PHF-tau-ir neurons, pattern I (without NFT and pattern II (with NFT, the distribution of which varies according to the cortical layer and area. Furthermore, the distribution of both GABAergic and glutamatergic terminals around the soma and proximal processes of PHF-tau-ir neurons does not seem to be altered as it is indistinguishable from both control cases and from adjacent neurons that did not contain PHF-tau. At the electron microscope level, a normal looking neuropil with typical symmetric and asymmetric synapses was observed around PHF-tau-ir neurons. These observations suggest that the synaptic connectivity around the perisomatic region of these PHF-tau-ir neurons was apparently unaltered.

LidiaBlazquez-Llorca

2010-06-01

364

Immunoreactivity and Radioimmunoscintigraphy of 4-Lysine Single Chain (Fv) Lym-1 Antibody for the Radiometal Chelation  

Energy Technology Data Exchange (ETDEWEB)

Small size of recombinant scFv, composed of VH and VL region of IgG, has many advantages such as faster blood clearance, improved tumor localization and reduced human anti-mouse antibody (HAMA) response. On the other hand, owing to small size, number of amino group, which was not involved in binding site, of ScFv lym-1 was insufficient in conjugation with CITC-DTPA chelator for radio metal labeling. The goal of this study is to introduce 4-lysine tag to the end of ScFv lym-1 sequence for radio metal conjugation and to evaluate the immunoreactivity and radioimmunoscintigraphy of chelator conjugated 4-lysine taq scFv lym-1 (4-lys scFv)

Jung, Jae Ho; Choi, Tae Hyun; Lee, Tae Sup; Uh, Gwang Sun; Chung, Wee Sup; Kim, Eun Jung; Cheon, Gi Jeong; Choi, Chang Woon; Lim, Sang Moo [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

2007-07-01

365

Immunoreactivity and Radioimmunoscintigraphy of 4-Lysine Single Chain (Fv) Lym-1 Antibody for the Radiometal Chelation  

International Nuclear Information System (INIS)

Small size of recombinant scFv, composed of VH and VL region of IgG, has many advantages such as faster blood clearance, improved tumor localization and reduced human anti-mouse antibody (HAMA) response. On the other hand, owing to small size, number of amino group, which was not involved in binding site, of ScFv lym-1 was insufficient in conjugation with CITC-DTPA chelator for radio metal labeling. The goal of this study is to introduce 4-lysine tag to the end of ScFv lym-1 sequence for radio metal conjugation and to evaluate the immunoreactivity and radioimmunoscintigraphy of chelator conjugated 4-lysine taq scFv lym-1 (4-lys scFv)

2007-05-10

366

Cogeneration of retrogradely labeled corticocortical projection and GABA-immunoreactive local circuit neurons in cerebral cortex.  

Science.gov (United States)

The times of origin of cortico-cortical projection neurons and local circuit neurons in rat visual cortex were determined. The birthdates of the projection neurons were assessed using a technique that combined retrograde labeling with lectin-bound horseradish peroxidase and tritiated thymidine autoradiography. The birthdates of some cortical local circuit neurons were determined by combining GABA immunocytochemistry with [3H]thymidine autoradiography. Double-labeled neurons (those with retrograde or immunoreactive label in their perikarya and autoradiographic silver grains over their nuclei) were born during the third week of gestation. Projection and local circuit neurons born on gestational day 14, 15, 17, 19 or 20 were located primarily in layer VIb, VIa, V, III or II, respectively. Thus, both populations of neurons are generated by parallel and concurrent inside-to-outside patterns. PMID:3910166

Miller, M W

1985-12-01

367

Immunoreactive atrial natriuretic factor is increased in ovine model of endotoxemia  

International Nuclear Information System (INIS)

A bolus of Escherichia coli endotoxin (1.5 ?g/kg) was administered to chronically instrumented sheep. Immunoreactive atrial natriuretic factor (IR-ANF) was measured in extracted plasma by radioimmunoassay. There was a thirteenfold increase in IR-ANF 2 h after endotoxin administration, and IR-ANF levels remained significantly elevated during the first 6 h. A marked diuresis and natriuresis occurred between 4 and 6 h. ANF not only affects renal function but is also associated with decreased cardiac output, increased peripheral resistance (in sheep), and decreased capillary absorption (in rats). These renal and hemodynamic changes are also characteristic of the early (first 6 h) response to endotoxin. Therefore ANF should be considered as a potential mediator of renal and hemodynamic changes induced by sepsis. It is difficult to determine if ANF elevation is an epiphenomenon or a causative factor, because no antagonist of ANF is currently available

1988-01-01

368

Immunoreactive atrial natriuretic factor is increased in ovine model of endotoxemia  

Energy Technology Data Exchange (ETDEWEB)

A bolus of Escherichia coli endotoxin (1.5 {mu}g/kg) was administered to chronically instrumented sheep. Immunoreactive atrial natriuretic factor (IR-ANF) was measured in extracted plasma by radioimmunoassay. There was a thirteenfold increase in IR-ANF 2 h after endotoxin administration, and IR-ANF levels remained significantly elevated during the first 6 h. A marked diuresis and natriuresis occurred between 4 and 6 h. ANF not only affects renal function but is also associated with decreased cardiac output, increased peripheral resistance (in sheep), and decreased capillary absorption (in rats). These renal and hemodynamic changes are also characteristic of the early (first 6 h) response to endotoxin. Therefore ANF should be considered as a potential mediator of renal and hemodynamic changes induced by sepsis. It is difficult to determine if ANF elevation is an epiphenomenon or a causative factor, because no antagonist of ANF is currently available.

Lubbesmeyer, H.J.; Woodson, L.; Traber, L.D.; Flynn, J.T.; Herndon, D.N.; Traber, D.L. (Univ. of Texas Medical Branch, Galveston (USA) Thomas Jefferson Medical College, Philadelphia, PA (USA) Westfaelian Wilhelms Univ., Muenster (West Germany))

1988-04-01

369

Immunoreactive forms of caldesmon in cultivated human vascular smooth muscle cells.  

Science.gov (United States)

150 kDa caldesmon was shown to be characteristic of vascular smooth muscle cells in normal tissue rather than in subculture. Subcultured smooth muscle cells from human aorta contained only the 70 kDa immunoreactive form of caldesmon. During the course of primary culture the amount of 150 kDa caldesmon as well as metavinculin decreased significantly whilst 70 kDa caldesmon became the predominant form, and by the onset of cell division the 150 kDa form was practically substituted by 70 kDa caldesmon. The data show that the predominance of 150 kDa caldesmon is characteristic of contractile smooth muscle cells, while in proliferating cells 70 kDa caldesmon is expressed. PMID:3109949

Glukhova, M A; Kabakov, A E; Ornatsky, O I; Vasilevskaya, T D; Koteliansky, V E; Smirnov, V N

1987-06-29

370

Decreased immunoreactivities of neocortical AMPA receptor subunits correlate with motor disability in Lewy body dementias.  

Science.gov (United States)

Dementia with Lewy bodies and Parkinson's disease dementia are different clinical phenotypes of Lewy body dementias differentiated by the temporal relationship between parkinsonism and dementia onset. At present, it is unclear whether the glutamatergic system is affected in these disorders. In this study, we measured ?-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor GluA subunits in the postmortem neocortex of a cohort of prospectively studied Lewy body dementia cases, as well as age-matched controls by immunoblotting. We found losses of GluA2/3/4 immunoreactivities in Lewy body dementias which correlated with higher pre-death Hoehn and Yahr scores and with longer Parkinson's disease duration before dementia onset, but not with dementia severity, cortical Lewy body burden, or amyloid plaque and neurofibrillary tangle burden. Our study suggests that GluA2/3/4 losses may be a neurochemical marker of motor disability in Lewy body dementias. PMID:23846847

Mohamed, Nur-Ezan; Howlett, David R; Ma, Lu; Francis, Paul T; Aarsland, Dag; Ballard, Clive G; McKeith, Ian G; Chen, Christopher P; Lai, Mitchell K P

2014-01-01

371

Plasma beta-endorphin-like immunoreactivity and its variations in baboons  

International Nuclear Information System (INIS)

This paper determines the level of beta-endorphin-like immunoreactivity (beta-elir) in the blood plasma of baboons and studies its changes in certain situations. For radioimmunoassay of beta-ELIR in the blood plasma, a standard kit and the appropriate technique were used. The background plasma beta-ELIR level of the baboons, in a state of quiet wakefulness, was 0.0 = 1.0 fmoles/ml. The total level of b-ELIR was 134 plus or minus 24 pg/ml. The data show that elevation of the plasma b-ELIR level accompanies stress formation, including the development of a state of shock in baboons. A definite role in the regulation of the plasma b-endorphin level may be played by the paraventricular-perifornical region of the hypothalamus

1986-04-01

372

Plasma levels of immunoreactive erythropoietin after acute blood loss in man  

Energy Technology Data Exchange (ETDEWEB)

Plasma levels of immunoreactive erythropoietin (Ep) were measured after acute blood loss (50, 75, 200 and 450 ml) in man in order to determine the volume of blood loss required to trigger an Ep response as well as to define the reticulocyte response. There was a highly significant (P<0.01) linear increase in reticulocyte count after 200 and 450 ml of blood loss. Analysis of trends also showed a highly significant (P<0.01) linear response of haematocrit and Ep after a 450 ml blood loss. The reticulocyte count increases were not dependent on a prior increase in Ep level indicating that at least two mechanisms are operative in restoring the red cell mass to normal after blood loss.

Miller, M.E. (Brookhaven National Lab., Upton, NY); Cronkite, E.P.; Garcia, J.F.

1982-01-01

373

Endogenous digoxin-like immunoreactive factor is elevated in advanced chronic respiratory failure.  

Science.gov (United States)

Digoxin-like immunoreactive factor (DLIF) is an endogenous substance with natriuretic and diuretic activity. Elevated plasma levels of DLIF are found in various clinical states characterized by water and sodium retention. Chronic respiratory failure, particularly of an advanced stage, also is frequently associated with water and sodium retention. In order to determine whether elevated plasma levels of DLIF are present in chronic respiratory failure, we measured plasma DLIF levels in seven patients (four with COPD [two of whom had associated sleep apnea disturbance] and three with kyphoscoliosis) suffering from advanced chronic respiratory failure with severe hypoxemia and hypercapnia. We found that in these patients plasma levels of DLIF were significantly higher than in healthy control subjects. We conclude that patients with advanced chronic respiratory failure respond with increased levels of DLIF. This may represent an attempt at homeostasis of water and sodium metabolism which is frequently deranged in this clinical condition. PMID:1309496

Varsano, S; Shilo, L; Bruderman, I; Dolev, S; Shenkman, L

1992-01-01

374

Digoxin-like immunoreactive factor(s) in human gonadotropin stimulated follicular fluid.  

Science.gov (United States)

Plasma digoxin-like immunoreactive factor(s) (DLIF) have been reported in various pathophysiological conditions associated with volume expansion and linked to the regulation of blood volume and pressure. We hypothesized that DLIF might be present in rapidly expanding gonadotropin-stimulated ovarian follicles. The mean total and free DLIF concentrations in the follicles (n = 9) studied were 4925 nmol/L and 1885 nmol/L, respectively. These concentrations were substantially higher than the plasma total and free DLIF levels in these women: 1216 nmol/L and 158 nmol/L, respectively (p less than 0.0001). The plasma DLIF levels in the gonadotropin-treated women were comparable to those in term pregnant women, which are known to be higher than those in non-pregnant women. The ovary thus may be a source of DLIF in the plasma of gonadotropin-treated women, and DLIF may have a role in ovarian follicular fluid homeostasis. PMID:2499591

Jakobi, P; Krivoy, N; Eibschitz, I; Ziskind, G; Barzilai, D; Paldi, E

1989-07-01

375

Fos-like immunoreactivity in locus coeruleus after classical conditioning of the rabbit's nictitating membrane response.  

Science.gov (United States)

We investigated the changes in Fos-like immunoreactivity in the locus coeruleus (LC) after classical conditioning of the rabbit's nictitating membrane. Specifically, we compared unpaired versus paired presentations of a tone conditioned stimulus (CS) and a tactile unconditioned stimulus (US; near the eye). After two training sessions, only paired presentations resulted in acquisition of a conditioned response. This was associated with comparatively less LC Fos expression than with unpaired presentations. Similar observations have been reported for the ventrolateral medulla which is a major source of afferents to LC. The present results are consistent with a role of LC in attention and learning: activity increases as the animal attends to the CS and US. When the relationship between CS and US has been established, LC activity decreases. PMID:9058416

Carrive, P; Kehoe, E J; Macrae, M; Paxinos, G

1997-02-14

376

Acute effect of ischemia on adrenomedullin immunoreactivity in the rat heart: an immunocytochemical study.  

Science.gov (United States)

We investigated by immunocytochemistry (ICC) the acute effects of ischemia on the distribution in the rat heart of adrenomedullin (AM), a potent hypotensive peptide which is expressed in the cardiovascular system, where it is known to play a major regulatory and protective role. Hearts, collected from adult male Sprague-Dawley rats, were perfused with the Langendorff technique, and "global" ischemia was obtained by stopping perfusion for 20 min. Hearts were frozen, and ICC was performed using a specific anti-rat AM1-50 antibody and secondary peroxidase-conjugated antibodies. ICC demonstrated AM-immunoreactivity (IR) in cardiomyocytes and especially in the wall of coronary vessels. Quantitative densitometry showed that acute ischemia significantly decreased AM-IR in coronary arterioles, thereby suggesting that it markedly stimulates AM release. The conclusion is drawn that acute ischemia and ensuing hypoxia activate in the rat heart the release of AM, which by its coronarodilatory action may enhance heart blood flow. PMID:15202018

Belloni, Anna S; Guidolin, Diego; Ceretta, Silvia; Bova, Sergio; Nussdorfer, Gastone G

2004-07-01

377

Basic FGF-like immunoreactivity in the developing and adult rat brainstem.  

Science.gov (United States)

Although a variety of in vitro and in vivo actions of basic fibroblast growth factor (bFGF) on neuronal cells have been documented, the physiological role of this protein in the nervous system is still contested. Since the distribution of a molecule in the nervous system may provide cues for an understanding of its possible roles, we have begun to study its cellular localization in the central and peripheral nervous system using immunocytochemistry with an anti-bFGF-specific antibody. Here we provide an account on the distribution of bFGF-like immunoreactivity (bFGF-IR) in the brainstem of the developing and adult rat. Basic FGF-IR was found to be widely distributed in motor and sensory nuclei. In all nuclei examined, only subpopulations of neurons were stained. Different staining patterns were found. For example, in the red nucleus weakly or unstained perikarya were surrounded by numerous immunoreactive fibers, often in close contact with the neuronal surface. In the reticular formation and facial nerve, many neuronal cell bodies showed a strong IR that extended into the processes. Glial cells were consistently unstained. During early postnatal development changes of the distribution of bFGF IR were found. From this wide distribution pattern of bFGF-IR, we conclude that bFGF may have more general and, possibly, diverse functions rather than a restricted role for a particular subset of neurons. Variations in the staining pattern of nerve cell bodies in a single nucleus may suggest a function related to neuronal activity. PMID:2026791

Grothe, C; Zachmann, K; Unsicker, K

1991-03-01

378

Decreased nucleotide excision repair in steatotic livers associates with myeloperoxidase-immunoreactivity  

International Nuclear Information System (INIS)

Chronic inflammation is characterized by the influx of neutrophils and is associated with an increased production of reactive oxygen species that can damage DNA. Oxidative DNA damage is generally thought to be involved in the increased risk of cancer in inflamed tissues. We previously demonstrated that activated neutrophil mediated oxidative stress results in a reduction in nucleotide excision repair (NER) capacity, which could further enhance mutagenesis. Inflammation and oxidative stress are critical factors in the progression of nonalcoholic fatty liver disease that is linked with enhanced liver cancer risk. In this report, we therefore evaluated the role of neutrophils and the associated oxidative stress in damage recognition and DNA repair in steatotic livers of 35 severely obese subjects with either nonalcoholic steatohepatitis (NASH) (n = 17) or steatosis alone (n = 18). The neutrophilic influx in liver was assessed by myeloperoxidase (MPO) staining and the amount of oxidative DNA damage by measuring M1dG adducts. No differences in M1dG adduct levels were observed between patients with or without NASH and also not between individuals with high or low MPO immunoreactivity. However, we found that high expression of MPO in the liver, irrespective of disease status, reduced the damage recognition capacity as determined by staining for histone 2AX phosphorylation (?H2AX). This reduction in ?H2AX formation in individuals with high MPO immunoreactivity was paralleled by a significant decrease in NER capacity as assessed by a functional repair assay, and was not related to cell proliferation. Thus, the observed reduction in NER capacity upon hepatic inflammation is associated with and may be a consequence of reduced damage recognition. These findings suggest a novel mechanism of liver cancer development in patients with nonalcoholic fatty liver disease.

2012-08-01

379

Putaminal upregulation of FosB/?FosB-like immunoreactivity in Parkinson's disease patients with dyskinesia.  

Science.gov (United States)

The transcription factor ?FosB is a mediator of maladaptive neuroplasticity in animal models of Parkinson's disease (PD) and L-DOPA-induced dyskinesia. Using an antibody that recognizes all known isoforms of FosB and ?FosB, we have examined the expression of these proteins in post-mortem basal ganglia sections from PD patients. The patient cases were classified as being dyskinetic or non-dyskinetic based on their clinical records. Sections from neurologically healthy controls were also included in the study. Compared to both controls and non-dyskinetic cases, the dyskinetic group showed a higher density of FosB/?FosB-immunopositive cells in the posterior putamen, which represents the motor region of the striatum in primates. In contrast, the number of FosB/?FosB-positive cells did not differ significantly among the groups in the caudate, a region primarily involved with the processing of cognitive and limbic-related information. Only sparse FosB/?FosB immunoreactivity was found in the in the pallidum externum and internum, and no significant group differences were detected in these nuclei. The putaminal elevation of FosB/?FosB-like immunoreactivity in patients who had been affected by L-DOPA-induced dyskinesia is consistent with results from both rat and non-human primate models of this movement disorder. The present findings support the hypothesis of an involvement of ?FosB-related transcription factors in the molecular mechanisms of L-DOPA-induced dyskinesia. PMID:23933656

Lindgren, H S; Rylander, D; Iderberg, H; Andersson, M; O'Sullivan, S S; Williams, D R; Lees, A J; Cenci, M A

2011-01-01

380

Decreased nucleotide excision repair in steatotic livers associates with myeloperoxidase-immunoreactivity  

Energy Technology Data Exchange (ETDEWEB)

Chronic inflammation is characterized by the influx of neutrophils and is associated with an increased production of reactive oxygen species that can damage DNA. Oxidative DNA damage is generally thought to be involved in the increased risk of cancer in inflamed tissues. We previously demonstrated that activated neutrophil mediated oxidative stress results in a reduction in nucleotide excision repair (NER) capacity, which could further enhance mutagenesis. Inflammation and oxidative stress are critical factors in the progression of nonalcoholic fatty liver disease that is linked with enhanced liver cancer risk. In this report, we therefore evaluated the role of neutrophils and the associated oxidative stress in damage recognition and DNA repair in steatotic livers of 35 severely obese subjects with either nonalcoholic steatohepatitis (NASH) (n = 17) or steatosis alone (n = 18). The neutrophilic influx in liver was assessed by myeloperoxidase (MPO) staining and the amount of oxidative DNA damage by measuring M{sub 1}dG adducts. No differences in M{sub 1}dG adduct levels were observed between patients with or without NASH and also not between individuals with high or low MPO immunoreactivity. However, we found that high expression of MPO in the liver, irrespective of disease status, reduced the damage recognition capacity as determined by staining for histone 2AX phosphorylation ({gamma}H2AX). This reduction in {gamma}H2AX formation in individuals with high MPO immunoreactivity was paralleled by a significant decrease in NER capacity as assessed by a functional repair assay, and was not related to cell proliferation. Thus, the observed reduction in NER capacity upon hepatic inflammation is associated with and may be a consequence of reduced damage recognition. These findings suggest a novel mechanism of liver cancer development in patients with nonalcoholic fatty liver disease.

Schults, Marten A.; Nagle, Peter W. [Department of Toxicology, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Rensen, Sander S. [Department of Surgery, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Godschalk, Roger W. [Department of Toxicology, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Munnia, Armelle; Peluso, Marco [Cancer Risk Factor Branch, ISPO Cancer Prevention and Research Institute, Via Cosimo il Vecchio 2, 50139 Florence (Italy); Claessen, Sandra M. [Department of Toxicogenomics, GROW-School for Oncology and Developmental Biology, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Greve, Jan W. [Department of Surgery, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Driessen, Ann [Department of Pathology, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Verdam, Froukje J.; Buurman, Wim A. [Department of Surgery, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Schooten, Frederik J. van [Department of Toxicology, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands); Chiu, Roland K., E-mail: r.k.chiu@med.umcg.nl [Department of Toxicology, NUTRIM-School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre, PO Box 616, 6200 MD Maastricht (Netherlands)

2012-08-01

 
 
 
 
381

Production of the recombinant single chain anti-B cell lymphoma antibody and evaluation of immunoreactivity  

Energy Technology Data Exchange (ETDEWEB)

Recombinant ScFv lym-1 was produced, using pET vector system for large scale production. ScFv lym-1 gene inserted pET-22b (+) vector, was expressed in E. coli BL-21 strain. ScFv lym-1 antibody extracted from periplasm, was purified with His-Taq column. To evaluated immunoreactivity with Raji cell, ScFv lym-1 was labeled with I-125 and I-125 ScFv lym-1 was purified with desalting column. Raji cell was injected into the C57BR/cdJ SCID mice. Gamma camera imaging were taken time point at 1, 8, 24 and 48 hr with 8 mm pinhole collimator. An active scFv lym-1 could be produced in E. coli with soluble from using pET vector system. Immunoreactivity and affinity constant of lgG lym-1 were 54% and 1.83 x 10{sup 9} M{sup -1}, respectively, and those of scFv lym-1 were 53.7% and 1.46 x 10{sup 9} M{sup -1}, respectively. Biodistribution of I-125 scFv lym-1 antibody showed faster clearance in blood, spleen, kidney and than I-125 lgG lym-1 antibody. Gamma camera image of I-125 scFv lym-1 antibody showed faster clearance and tumor targeting liver than I-125 lgG lym-1 antibody. In vitro properties of scFv lym-1 were similar to those of lgG lym-1. ScFv lym-1 showed faster blood clearance than lgG lym-1. These results suggest that scFv lym-1 antibody can be useful for tumor imaging agent.

Jung, Jae Ho; Choi, Tae Hyun; Woo, Kang Sun; Chung, Wee Sup; Kim, Soo Gwan; Cheon, Gi Jeong; Choi, Chang Woon; Lim, Sang Moo [Korea Institute of Radiological and Medical Science, Seoul (Korea, Republic of)

2006-08-15

382

Electron microscopic examination of the endomorphin 2-like immunoreactive neurons in the rat hypothalamus.  

Science.gov (United States)

Endomorphins are endogenous opioid peptides with high affinity and selectivity for the mu-opioid receptor. In the present study, we examined the morphology of the endomorphin 2-like immunoreactive (EM2-LI) neurons in the hypothalamus at the light and electron microscopic levels. At the light microscopic level, EM2-LI neurons were found mostly distributed in the regions between the dorsomedial and ventromedial hypothalamic nuclei and the region near the third ventricle. At the electron microscopic level, EM2-LI perikarya could be divided into two groups. Type I perikarya contained relatively undeveloped endoplasmic reticulum and Golgi apparatus while type II perikarya contained well-developed rough-surfaced endoplasmic reticulum and Golgi apparatus. Both type I and type II neurons contained numerous EM2-LI dense-cored vesicles. Type II perikarya and dendrites received synapses and showed immunoreactivity in the endoplasmic reticulum and Golgi apparatus. EM2-LI axon terminals formed synapses with both immunonegative and immunopositive dendrites. In some cases, the axon terminals contained both immunonegative and immunopositive dense-cored vesicles. EM2-LI neurons often had synaptic relationships with neurons containing immunonegative dense-cored vesicles. Myelinated axon shafts containing EM2-LI were also found. This first demonstration of the ultrastructure and synaptic relationships of EM2-LI neurons in the hypothalamus provides morphological evidence that suggests (1) endomorphin 2-containing neurons modulate physiological function through synaptic relationships; (2) endomorphin 2 may coexist with other neurotransmitters in the same neurons; and (3) endomorphin 2-containing neurons could modulate other endomorphin 2-containing neurons as well as those containing other neurotransmitters. PMID:12676373

Wang, Q-P; Zadina, J E; Guan, J-L; Kastin, A J; Shioda, S

2003-04-18

383

Identification of initially appearing glycine-immunoreactive neurons in the embryonic zebrafish brain.  

Science.gov (United States)

Glycine is a major inhibitory neurotransmitter in the central nervous system of vertebrates. Here, we report the initial development of glycine-immunoreactive (Gly-ir) neurons and fibers in zebrafish. The earliest Gly-ir cells were found in the hindbrain and rostral spinal cord by 20 h post-fertilization (hpf). Gly-ir cells in rhombomeres 5 and 6 that also expressed glycine transporter 2 (glyt2) mRNA were highly stereotyped; they were bilaterally located and their axons ran across the midline and gradually turned caudally, joining the medial longitudinal fascicles in the spinal cord by 24 hpf. Gly-ir neurons in rhombomere 5 were uniquely identified, since there was one per hemisegment, whereas the number of Gly-ir neurons in rhombomere 6 were variable from one to three per hemisegment. Labeling of these neurons by single-cell electroporation and tracing them until the larval stage revealed that they became MiD2cm and MiD3cm, respectively. The retrograde labeling of reticulo-spinal neurons in Tg(glyt2:gfp) larva, which express GFP in Gly-ir cells, and a genetic mosaic analysis with glyt2:gfp DNA construct also supported this notion. Gly-ir cells were also distributed widely in the anterior brain by 27 hpf, whereas glyt2 was hardly expressed. Double staining with anti-glycine and anti-GABA antibodies demonstrated distinct distributions of Gly-ir and GABA-ir cells, as well as the presence of doubly immunoreactive cells in the brain and placodes. These results provide evidence of identifiable glycinergic (Gly-ir/glyt2-positive) neurons in vertebrate embryos, and they can be used in further studies of the neurons' development and function at the single-cell level. © 2013 Wiley Periodicals, Inc. Develop Neurobiol 74: 616-632, 2014. PMID:24318965

Moly, Pricila Khan; Ikenaga, Takanori; Kamihagi, Chihiro; Islam, A F M Tariqul; Hatta, Kohei

2014-06-01

384

Substance P Immunoreactivity Exhibits Frequent Colocalization with Kisspeptin and Neurokinin B in the Human Infundibular Region  

Science.gov (United States)

Neurons synthesizing neurokinin B (NKB) and kisspeptin (KP) in the hypothalamic arcuate nucleus represent important upstream regulators of pulsatile gonadotropin-releasing hormone (GnRH) neurosecretion. In search of neuropeptides co-expressed in analogous neurons of the human infundibular nucleus (Inf), we have carried out immunohistochemical studies of the tachykinin peptide Substance P (SP) in autopsy samples from men (21-78 years) and postmenopausal (53-83 years) women. Significantly higher numbers of SP-immunoreactive (IR) neurons and darker labeling were observed in the Inf of postmenopausal women than in age-matched men. Triple-immunofluorescent studies localized SP immunoreactivity to considerable subsets of KP-IR and NKB-IR axons and perikarya in the infundibular region. In postmenopausal women, 25.1% of NKB-IR and 30.6% of KP-IR perikarya contained SP and 16.5% of all immunolabeled cell bodies were triple-labeled. Triple-, double- and single-labeled SP-IR axons innervated densely the portal capillaries of the infundibular stalk. In quadruple-labeled sections, these axons formed occasional contacts with GnRH-IR axons. Presence of SP in NKB and KP neurons increases the functional complexity of the putative pulse generator network. First, it is possible that SP modulates the effects of KP and NKB in axo-somatic and axo-dendritic afferents to GnRH neurons. Intrinsic SP may also affect the activity and/or neuropeptide release of NKB and KP neurons via autocrine/paracrine actions. In the infundibular stalk, SP may influence the KP and NKB secretory output via additional autocrine/paracrine mechanisms or regulate GnRH neurosecretion directly. Finally, possible co-release of SP with KP and NKB into the portal circulation could underlie further actions on adenohypophysial gonadotrophs.

Hrabovszky, Erik; Borsay, Beata A.; Racz, Kalman; Herczeg, Laszlo; Ciofi, Philippe; Bloom, Stephen R.; Ghatei, Mohammad A.; Dhillo, Waljit S.; Liposits, Zsolt

2013-01-01

385

Microalbuminuria, indicated by total versus immunoreactive urinary albumins, in acute ischemic stroke patients.  

Science.gov (United States)

Microalbuminuria, assessed by measuring immunoreactive albumin, is common in patients with cerebrovascular disease and is associated with increased risk of stroke. Total urinary albumin (t-uAlb) comprises both immunoreactive albumin (ir-uAlb) and nonimmunoreactive albumin (nir-uAlb). We hypothesized that t-uAlb is a more sensitive indicator of microalbuminuria than ir-uAlb, and that measurement of t-uAlb will increase the prevalence of microalbuminuria in ischemic stroke patients compared with measurement of ir-uAlb and will show a stronger correlation with the severity of stroke and oxidative stress. In urine samples from 98 patients with ischemic stroke, the albumin-to-creatinine ratios t-uAlb/uCreat and ir-uAlb/uCreat were measured by high-performance liquid chromatography (HPLC) and immunoturbidimetry (IT), and the nir-uAlb/uCreat ratio was calculated. Urinary ortho-tyrosine (o-Tyr/uCreat), an indicator of oxidative stress, was measured by HPLC. The severity of stroke was scored based on the National Institutes of Health Stroke Scale (NIHSS). The prevalence of microalbuminuria detected by HPLC was significantly higher than that detetcted by IT (66.3 vs 36.7%). Although all forms of albumin showed significant correlation with stroke severity (t-uAlb: r = 0.24, P stroke severity (B = 0.20, ? = 0.35, P .05). Our data suggest that in acute ischemic stroke patients, t-uAlb is a more sensitive indicator of microalbuminuria than the presently used ir-uAlb. Future studies should aim to elucidate the underlying mechanisms for the relationship among urinary albumins and cerebrovascular diseases and the role of urinary albumins in risk stratification for stroke. PMID:20813547

Toth, Peter; Koller, Akos; Pusch, Gabriella; Bosnyak, Edit; Szapary, Laszlo; Komoly, Samuel; Marko, Lajos; Nagy, Judit; Wittmann, Istvan

2011-11-01

386

Production of the recombinant single chain anti-B cell lymphoma antibody and evaluation of immunoreactivity  

International Nuclear Information System (INIS)

Recombinant ScFv lym-1 was produced, using pET vector system for large scale production. ScFv lym-1 gene inserted pET-22b (+) vector, was expressed in E. coli BL-21 strain. ScFv lym-1 antibody extracted from periplasm, was purified with His-Taq column. To evaluated immunoreactivity with Raji cell, ScFv lym-1 was labeled with I-125 and I-125 ScFv lym-1 was purified with desalting column. Raji cell was injected into the C57BR/cdJ SCID mice. Gamma camera imaging were taken time point at 1, 8, 24 and 48 hr with 8 mm pinhole collimator. An active scFv lym-1 could be produced in E. coli with soluble from using pET vector system. Immunoreactivity and affinity constant of lgG lym-1 were 54% and 1.83 x 109 M-1, respectively, and those of scFv lym-1 were 53.7% and 1.46 x 109 M-1, respectively. Biodistribution of I-125 scFv lym-1 antibody showed faster clearance in blood, spleen, kidney and than I-125 lgG lym-1 antibody. Gamma camera image of I-125 scFv lym-1 antibody showed faster clearance and tumor targeting liver than I-125 lgG lym-1 antibody. In vitro properties of scFv lym-1 were similar to those of lgG lym-1. ScFv lym-1 showed faster blood clearance than lgG lym-1. These results suggest that scFv lym-1 antibody can be useful for tumor imaging agent

2006-08-01

387

Loss of Calbindin immunoreactivity in the dentate gyrus distinguishes Alzheimer's disease from other neurodegenerative dementias.  

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Calbindin (Cb) is one of the major Ca(2+) binding proteins exhibiting neuromodulatory functions such as long-term potentiation (LTP), synaptic plasticity, and memory functions. It is expressed in hippocampal interneurons, pyramidal cells and granule cells of the dentate gyrus (DGCs). Cb mRNA levels remain stable during normal ageing, but decrease in Alzheimer's, Huntington, and Parkinson's disease. A recent study suggested a link between A?-induced Alzheimer's disease (AD)-related cognitive deficits and neuronal depletion of Cb. To evaluate whether this is specific for AD, we performed a comparative study of Cb immunoreactivity of DGCs in cases with AD-related neuropathologic change (49), grouped according to the stages of Braak and Braak, BB), Creutzfeldt-Jakob-disease (16), FTLD-tau Pick's disease type (PiD; 5), argyrophilic grain disease (8), and FTLD-TDP types A and B (6). The group of AD cases with BB stages V and VI showed the highest proportion of Cb negative cells in the DGC when compared to all other groups except PiD. The ratio of negative cells correlated significantly with the BB stages. While the total number of DGCs decreased with age in our series, loss of Cb immunoreactivity was shown to be age-dependent only in PiD and FTLD-TDP. We conclude, that late stage AD-neuropathologic change (BB V and VI stages) associates with significantly higher ratios of Cb negative DGCs and this correlates with advanced BB stage. This might suggest an accumulative effect of an epilepsy-