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Abnormal regenerative responses and impaired axonal outgrowth after nerve crush in TDP-43 transgenic mouse models of amyotrophic lateral sclerosis.  

Science.gov (United States)

Tar DNA binding protein 43 (TDP-43) mislocalization and aggregation is a hallmark of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar dementia. Moreover, TDP-43 mRNA was found to be upregulated by ?2.5-fold in the spinal cord of sporadic ALS subjects. Here we have examined the effects of nerve injury in new transgenic mouse models overexpressing by approximately threefold wild-type or mutant (G348C) TDP-43 species. Four weeks after axonal crush of sciatic nerve, TDP-43 transgenic mice remained paralyzed at the injured limb unlike control mice, which had regained most of their normal mobility. In contrast to normal mice, TDP-43 transgenic mice exhibited sustained elevation of TDP-43 cytoplasmic levels in motor neurons after nerve crush, and the relocalization of TDP-43 to the nucleus was delayed by several weeks. After crush, peripherin and ubiquitin levels remained also significantly elevated in TDP-43 transgenic mice compared with control mice. Analysis of the sciatic nerve at 11 d after nerve crush showed that the number of regenerating axons in the distal portion of the lesion was considerably reduced in TDP-43 transgenic mice, especially in TDP-43(G348C) mice, which exhibited a reduction of ?40%. In addition, markers of neuroinflammation were detected at much higher levels in TDP-43 transgenic mice. These results suggest that a deregulation of TDP-43 expression in ALS is a phenomenon that can affect the regenerative responses to neuronal injury and regrowth potential of axons. PMID:23238732

Swarup, Vivek; Audet, Jean-Nicolas; Phaneuf, Daniel; Kriz, Jasna; Julien, Jean-Pierre

2012-12-12

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Abnormal regenerative responses and impaired axonal outgrowth after nerve crush in TDP-43 transgenic mouse models of amyotrophic lateral sclerosis.  

UK PubMed Central (United Kingdom)

Tar DNA binding protein 43 (TDP-43) mislocalization and aggregation is a hallmark of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar dementia. Moreover, TDP-43 mRNA was found to be upregulated by ?2.5-fold in the spinal cord of sporadic ALS subjects. Here we have examined the effects of nerve injury in new transgenic mouse models overexpressing by approximately threefold wild-type or mutant (G348C) TDP-43 species. Four weeks after axonal crush of sciatic nerve, TDP-43 transgenic mice remained paralyzed at the injured limb unlike control mice, which had regained most of their normal mobility. In contrast to normal mice, TDP-43 transgenic mice exhibited sustained elevation of TDP-43 cytoplasmic levels in motor neurons after nerve crush, and the relocalization of TDP-43 to the nucleus was delayed by several weeks. After crush, peripherin and ubiquitin levels remained also significantly elevated in TDP-43 transgenic mice compared with control mice. Analysis of the sciatic nerve at 11 d after nerve crush showed that the number of regenerating axons in the distal portion of the lesion was considerably reduced in TDP-43 transgenic mice, especially in TDP-43(G348C) mice, which exhibited a reduction of ?40%. In addition, markers of neuroinflammation were detected at much higher levels in TDP-43 transgenic mice. These results suggest that a deregulation of TDP-43 expression in ALS is a phenomenon that can affect the regenerative responses to neuronal injury and regrowth potential of axons.

Swarup V; Audet JN; Phaneuf D; Kriz J; Julien JP

2012-12-01

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Asparaginyl endopeptidase cleaves TDP-43 in brain.  

UK PubMed Central (United Kingdom)

TAR DNA-binding protein 43 (TDP-43) is a nuclear protein involved in RNA splicing and a major protein component in ubiquitin-positive, tau-negative inclusions of frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Under disease conditions, TDP-43 redistributes to the cytoplasm where it can be phosphorylated, ubiquitinated, and proteolytically cleaved. Enzymes responsible for TDP-43 proteolytic processing in brain remain largely unreported. Using a MS approach, we identified two truncated TDP-43 peptides, terminating C-terminal to asparagines 291 (N291) and 306 (N306). The only documented mammalian enzyme capable of cleaving C-terminal to asparagine is asparaginyl endopeptidase (AEP). TDP-43-immunoreactive fragments (~35 and 32 kDa) predicted to be generated by AEP cleavage at N291 and N306 were observed by Western blot analyses of postmortem frontotemporal lobar degeneration brain tissue and cultured human cells over-expressing TDP-43. Studies in vitro determined that AEP can directly cleave TDP-43 at seven sites, including N291 and N306. Western blots of brain homogenates isolated from AEP-null mice and wild-type littermate controls revealed that TDP-43 proteolytic fragments were substantially reduced in the absence of AEP in vivo. Taken together, we conclude that TDP-43 is cleaved by AEP in brain. Moreover, these data highlight the utility of combining proteomic strategies in vitro and in vivo to provide insight into TDP-43 biology that will fuel the design of more detailed models of disease pathogenesis.

Herskowitz JH; Gozal YM; Duong DM; Dammer EB; Gearing M; Ye K; Lah JJ; Peng J; Levey AI; Seyfried NT

2012-08-01

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

UK PubMed Central (United Kingdom)

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.

Xu YF; Prudencio M; Hubbard JM; Tong J; Whitelaw EC; Jansen-West K; Stetler C; Cao X; Song J; Zhang YJ

2013-01-01

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The Pathological Phenotypes of Human TDP-43 Transgenic Mouse Models Are Independent of Downregulation of Mouse Tdp-43  

Science.gov (United States)

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.

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

UK PubMed Central (United Kingdom)

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 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 (p.M337V, p.N345K, and p.I383V) in familial ALS patients, two of which were novel. With over 40 TARDBP mutations now discovered, there exists conclusive evidence that TDP-43 plays a direct role in neurodegeneration. The onus is now on researchers to elucidate the mechanisms by which mutant TDP-43 confers toxicity, and to exploit these findings to gain a better understanding of how TDP-43 contributes to the pathogenesis of disease. Our biochemical analysis of TDP-43 in ALS patient lymphoblastoid cell lines revealed a substantial increase in TDP-43 truncation products, including a ? 25 kDa fragment, compared to control lymphoblastoid cell lines. We discuss the putative harmful consequence of abnormal TDP-43 fragmentation, as well as highlight additional mechanisms of toxicity associated with mutant TDP-43.

Gendron TF; Rademakers R; Petrucelli L

2013-01-01

7

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 deve...

Cannon, Ashley; Yang, Baoli; Knight, Joshua; Farnham, Ian M.; Zhang, Yongjie; Wuertzer, Charles A.; D’Alton, Simon

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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.  

UK PubMed Central (United Kingdom)

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 found that a 2-5-fold increase in TDP-43 expression over the endogenous level induced death of NSC34 motor neuronal cells and primary cortical neurons. TDP-43-induced death is associated with up-regulation of Bim expression and down-regulation of Bcl-xL expression. siRNA-mediated reduction of Bim expression attenuates TDP-43-induced death. Accumulated evidence indicates that caspases are activated in neurons of ALS and FTLD-U patients, and activated caspase-mediated cleavage of TDP-43 generates CTFs of TDP-43. Here, we further found that the ER (endoplasmic reticulum) stress- or staurosporine-mediated activation of caspases leads to cleavage of TDP-43 at Asp(89) and Asp(169), generating CTF35 (TDP-43-(90-414)) and CTF27 (TDP-43-(170-414)) in cultured neuronal cells. In contrast to TDP-43, CTF27 is unable to induce death while it forms aggregates. CTF35 was weaker than full-length TDP-43 in inducing death. A cleavage-resistant mutant of TDP-43 (TDP-43-D89E/D169E) showed stronger death-inducing activity than wild-type TDP-43. These results suggest that disease-related activation of caspases may attenuate TDP-43-induced toxicity by promoting TDP-43 cleavage.

Suzuki H; Lee K; Matsuoka M

2011-04-01

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

Directory of Open Access Journals (Sweden)

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.

Xu Ya-Fei; Zhang Yong-Jie; Lin Wen-Lang; Cao Xiangkun; Stetler Caroline; Dickson Dennis W; Lewis Jada; Petrucelli Leonard

2011-01-01

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

UK PubMed Central (United Kingdom)

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 disorders; however, unknown environmental and genetic factors may regulate this TDP-43 pathology. Using transgenic mouse models of several diseases of the central nervous system, we explored whether a primary proteinopathy might secondarily drive TDP-43 proteinopathy. We found abnormal, cytoplasmic accumulation of phosphorylated TDP-43 specifically in two tau transgenic models, but TDP-43 pathology was absent in mouse models of A? deposition, ?-synucleinopathy or Huntington's disease. Though tau pathology showed considerable overlap with cytoplasmic, phosphorylated TDP-43, tau pathology generally preceded TDP-43 pathology. Biochemical analysis confirmed the presence of TDP-43 abnormalities in the tau mice, which showed increased levels of high molecular weight, soluble TDP-43 and insoluble full-length and ~35 kD TDP-43. These data demonstrate that the neurodegenerative cascade associated with a primary tauopathy in tau transgenic mice can also promote TDP-43 abnormalities. These findings provide the first in vivo models to understand how TDP-43 pathology may arise as a secondary consequence of a primary proteinopathy.

Clippinger AK; D'Alton S; Lin WL; Gendron TF; Howard J; Borchelt DR; Cannon A; Carlomagno Y; Chakrabarty P; Cook C; Golde TE; Levites Y; Ranum L; Schultheis PJ; Xu G; Petrucelli L; Sahara N; Dickson DW; Giasson B; Lewis J

2013-07-01

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Cytoplasmic Inclusions of TDP-43 in Neurodegenerative Diseases: A Potential Role for Caspases  

Science.gov (United States)

TAR DNA-binding protein-43 (TDP-43) proteinopathies are classified based upon the extent of modified TDP-43 inclusions and include a growing number of neurodegenerative diseases including amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration with ubiquitin immunoreactive, tau negative inclusions (FTLD-U) and FTLD with motor neuron disease (FTLD-MND). In addition, TDP-43 inclusions have also been identified in a number of other neurodegenerative disorders including Alzheimer's disease, corticobasal degeneration, Lewy body related diseases and Pick's disease. Current understanding suggests that in these diseases, TDP-43 is relocated from the nucleus to the cytoplasm and sequestered into inclusions that contain modified TDP-43. Major modifications of TDP-43 have been identified as being hyperphosphorylation and proteolytic cleavage by caspases. In this review a summary of the major findings regarding the proteolytic modification of TDP-43 will be discussed as well as potential toxic-gain mechanisms these fragments may cause including cytoskeletal disruptions.

Rohn, Troy T.

2009-01-01

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The ALS disease associated mutant TDP-43 impairs mitochondrial dynamics and function in motor neurons.  

UK PubMed Central (United Kingdom)

Mutations in TDP-43 lead to familial ALS. Expanding evidence suggests that impaired mitochondrial dynamics likely contribute to the selective degeneration of motor neurons in SOD1-associated ALS. In this study, we investigated whether and how TDP-43 mutations might impact mitochondrial dynamics and function. We demonstrated that overexpression of wild type TDP-43 resulted in reduced mitochondrial length and density in neurites of primary motor neurons, features further exacerbated by ALS associated TDP-43 mutants Q331 K and M337V. In contrast, suppression of TDP-43 resulted in significantly increased mitochondrial length and density in neurites, suggesting a specific role of TDP-43 in regulating mitochondrial dynamics. Surprisingly, both TDP-43 overexpression and suppression impaired mitochondrial movement. We further showed that abnormal localization of TDP-43 in cytoplasm induced substantial and widespread abnormal mitochondrial dynamics. TDP-43 co-localized with mitochondria in motor neurons and their colocalization was enhanced by ALS associated mutant. Importantly, co-expression of mitochondrial fusion protein mitofusin 2 (Mfn2) could abolish TDP-43 induced mitochondrial dynamics abnormalities and mitochondrial dysfunction. Taken together, this data suggests that mutant TDP-43 impairs mitochondrial dynamics through enhanced localization on mitochondria, which causes mitochondrial dysfunction. Therefore, abnormal mitochondrial dynamics is likely a common feature of ALS which could be potential new therapeutic targets to treat ALS.

Wang W; Li L; Lin WL; Dickson DW; Petrucelli L; Zhang T; Wang X

2013-07-01

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TDP-43 regulates its mRNA levels through a negative feedback loop  

Science.gov (United States)

TAR DNA-binding protein (TDP-43) is an evolutionarily conserved heterogeneous nuclear ribonucleoprotein (hnRNP) involved in RNA processing, whose abnormal cellular distribution and post-translational modification are key markers of certain neurodegenerative diseases, such as amyotrophic lateral sclerosis and frontotemporal lobar degeneration. We generated human cell lines expressing tagged forms of wild-type and mutant TDP-43 and observed that TDP-43 controls its own expression through a negative feedback loop. The RNA-binding properties of TDP-43 are essential for the autoregulatory activity through binding to 3? UTR sequences in its own mRNA. Our analysis indicated that the C-terminal region of TDP-43, which mediates TDP-43–hnRNP interactions, is also required for self-regulation. TDP-43 binding to its 3? UTR does not significantly change the pre-mRNA splicing pattern but promotes RNA instability. Moreover, blocking exosome-mediated degradation partially recovers TDP-43 levels. Our findings demonstrate that cellular TDP-43 levels are under tight control and it is likely that disease-associated TDP-43 aggregates disrupt TDP-43 self-regulation, thus contributing to pathogenesis.

Ayala, Youhna M; De Conti, Laura; Avendano-Vazquez, S Erendira; Dhir, Ashish; Romano, Maurizio; D'Ambrogio, Andrea; Tollervey, James; Ule, Jernej; Baralle, Marco; Buratti, Emanuele; Baralle, Francisco E

2011-01-01

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RNP2 of RNA recognition motif 1 plays a central role in the aberrant modification of TDP-43.  

UK PubMed Central (United Kingdom)

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 uncertain. Here we report that the 32 kDa C-terminal fragment of TDP-43, which lacks the RNP2 motif of RNA binding motif 1 (RRM1), formed aggregates in cultured cells, and that similar phenotypes were obtained when the RNP2 motif was either deleted from or mutated in full-length TDP-43. These aggregations were ubiquitinated, phosphorylated and truncated, and sequestered the 25 kDa C-terminal TDP-43 fragment seen in the neurons of TDP-43 proteinopathy patients. In addition, incubation with RNase decreased the solubility of TDP-43 in cell lysates. These findings suggest that the RNP2 motif of RRM1 plays a substantial role in pathological TDP-43 modifications and that it is possible that disruption of RNA binding may underlie the process of TDP-43 aggregation.

Takagi S; Iguchi Y; Katsuno M; Ishigaki S; Ikenaka K; Fujioka Y; Honda D; Niwa J; Tanaka F; Watanabe H; Adachi H; Sobue G

2013-01-01

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TDP-43 expression in mouse models of amyotrophic lateral sclerosis and spinal muscular atrophy  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Redistribution of nuclear TAR DNA binding protein 43 (TDP-43) to the cytoplasm and ubiquitinated inclusions of spinal motor neurons and glial cells is characteristic of amyotrophic lateral sclerosis (ALS) pathology. Recent evidence suggests that TDP-43 pathology is common to sporadic ALS and familial ALS without SOD1 mutation, but not SOD1-related fALS cases. Furthermore, it remains unclear whether TDP-43 abnormalities occur in non-ALS forms of motor neuron disease. Here, we characterise TDP-43 localisation, expression levels and post-translational modifications in mouse models of ALS and spinal muscular atrophy (SMA). Results TDP-43 mislocalisation to ubiquitinated inclusions or cytoplasm was notably lacking in anterior horn cells from transgenic mutant SOD1G93A mice. In addition, abnormally phosphorylated or truncated TDP-43 species were not detected in fractionated ALS mouse spinal cord or brain. Despite partial colocalisation of TDP-43 with SMN, depletion of SMN- and coilin-positive Cajal bodies in motor neurons of affected SMA mice did not alter nuclear TDP-43 distribution, expression or biochemistry in spinal cords. Conclusion These results emphasise that TDP-43 pathology characteristic of human sporadic ALS is not a core component of the neurodegenerative mechanisms caused by SOD1 mutation or SMN deficiency in mouse models of ALS and SMA, respectively.

Turner Bradley J; Bäumer Dirk; Parkinson Nicholas J; Scaber Jakub; Ansorge Olaf; Talbot Kevin

2008-01-01

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RNA binding mediates neurotoxicity in the transgenic Drosophila model of TDP-43 proteinopathy.  

UK PubMed Central (United Kingdom)

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive and selective loss of motor neurons. The discovery of mutations in the gene encoding an RNA binding protein, TAR DNA-binding protein of 43 kDa (TDP-43), in familial ALS strongly implicated abnormalities in RNA processing in the pathogenesis of ALS, although the mechanisms whereby TDP-43 leads to neurodegeneration remain elusive. To clarify the mechanism of degeneration caused by TDP-43, we generated transgenic Drosophila melanogaster expressing a series of systematically modified human TDP-43 genes in the retinal photoreceptor neurons. Overexpression of wild-type TDP-43 resulted in vacuolar degeneration of the photoreceptor neurons associated with thinning of the retina, which was significantly exacerbated by mutations of TDP-43 linked to familial ALS or disrupting its nuclear localization signal. Remarkably, these degenerative phenotypes were completely normalized by addition of a mutation or deletion of the RNA recognition motif that abolishes the RNA binding ability of TDP-43. Altogether, our results suggest that RNA-binding is key to the neurodegeneration caused by overexpression of TDP-43, and that abnormalities in RNA processing may be crucial to the pathogenesis of TDP-43 proteinopathy.

Ihara R; Matsukawa K; Nagata Y; Kunugi H; Tsuji S; Chihara T; Kuranaga E; Miura M; Wakabayashi T; Hashimoto T; Iwatsubo T

2013-06-01

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ALS/FTLD-linked TDP-43 regulates neurite morphology and cell survival in differentiated neurons.  

UK PubMed Central (United Kingdom)

Tar-DNA binding protein of 43kDa (TDP-43) has been characterized as a major component of protein aggregates in brains with neurodegenerative diseases such as frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). However, physiological roles of TDP-43 and early cellular pathogenic effects caused by disease associated mutations in differentiated neurons are still largely unknown. Here, we investigated the physiological roles of TDP-43 and the effects of missense mutations associated with diseases in differentiated cortical neurons. The reduction of TDP-43 by siRNA increased abnormal neurites and decreased cell viability. ALS/FTLD-associated missense mutant proteins (A315T, Q331K, and M337V) were partially mislocalized to the cytosol and neurites when compared to wild-type and showed abnormal neurites similar to those observed in cases of loss of TDP-43. Interestingly, cytosolic expression of wild-type TDP-43 with mutated nuclear localization signals also induced abnormal neurtie morphology and reduction of cell viability. However, there was no significant difference in the effects of cytosolic expression in neuronal morphology and cell toxicity between wild-type and missense mutant proteins. Thus, our results suggest that mislocalization of missense mutant TDP-43 may contribute to loss of TDP-43 function and affect neuronal morphology, probably via dominant negative action before severe neurodegeneration in differentiated cortical neurons.

Han JH; Yu TH; Ryu HH; Jun MH; Ban BK; Jang DJ; Lee JA

2013-08-01

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TDP-43 is recruited to stress granules in conditions of oxidative insult.  

UK PubMed Central (United Kingdom)

Transactive response DNA-binding protein 43 (TDP-43) forms abnormal ubiquitinated and phosphorylated inclusions in brain tissues from patients with amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration. TDP-43 is a DNA/RNA-binding protein involved in RNA processing, such as transcription, pre-mRNA splicing, mRNA stabilization and transport to dendrites. We found that in response to oxidative stress and to environmental insults of different types TDP-43 is capable to assemble into stress granules (SGs), ribonucleoprotein complexes where protein synthesis is temporarily arrested. We demonstrated that a specific aminoacidic interval (216-315) in the C-terminal region and the RNA-recognition motif 1 domain are both implicated in TDP-43 participation in SGs as their deletion prevented the recruitment of TDP-43 into SGs. Our data show that TDP-43 is a specific component of SGs and not of processing bodies, although we proved that TDP-43 is not necessary for SG formation, and its gene silencing does not impair cell survival during stress. The analysis of spinal cord tissue from ALS patients showed that SG markers are not entrapped in TDP-43 pathological inclusions. Although SGs were not evident in ALS brains, we speculate that an altered control of mRNA translation in stressful conditions may trigger motor neuron degeneration at early stages of the disease.

Colombrita C; Zennaro E; Fallini C; Weber M; Sommacal A; Buratti E; Silani V; Ratti A

2009-11-01

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TDP-43 is recruited to stress granules in conditions of oxidative insult.  

Science.gov (United States)

Transactive response DNA-binding protein 43 (TDP-43) forms abnormal ubiquitinated and phosphorylated inclusions in brain tissues from patients with amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration. TDP-43 is a DNA/RNA-binding protein involved in RNA processing, such as transcription, pre-mRNA splicing, mRNA stabilization and transport to dendrites. We found that in response to oxidative stress and to environmental insults of different types TDP-43 is capable to assemble into stress granules (SGs), ribonucleoprotein complexes where protein synthesis is temporarily arrested. We demonstrated that a specific aminoacidic interval (216-315) in the C-terminal region and the RNA-recognition motif 1 domain are both implicated in TDP-43 participation in SGs as their deletion prevented the recruitment of TDP-43 into SGs. Our data show that TDP-43 is a specific component of SGs and not of processing bodies, although we proved that TDP-43 is not necessary for SG formation, and its gene silencing does not impair cell survival during stress. The analysis of spinal cord tissue from ALS patients showed that SG markers are not entrapped in TDP-43 pathological inclusions. Although SGs were not evident in ALS brains, we speculate that an altered control of mRNA translation in stressful conditions may trigger motor neuron degeneration at early stages of the disease. PMID:19765185

Colombrita, Claudia; Zennaro, Eleonora; Fallini, Claudia; Weber, Markus; Sommacal, Andreas; Buratti, Emanuele; Silani, Vincenzo; Ratti, Antonia

2009-09-16

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Reduced expression of BTBD10 in anterior horn cells with Golgi fragmentation and pTDP-43-positive inclusions in patients with sporadic amyotrophic lateral sclerosis.  

UK PubMed Central (United Kingdom)

Overexpression of BTBD10 (BTB/POZ domain-containing protein 10) suppresses G93A-superoxide dismutase 1 (SOD1)-induced motor neuron death in a cell-based amyotrophic lateral sclerosis (ALS) model. In the present study, paraffin sections of spinal cords from 13 patients with sporadic ALS and 10 with non-ALS disorders were immunostained using a polyclonal anti-BTBD10 antibody. Reduced BTBD10 expression in the anterior horn cells was more frequent in spinal cords from ALS patients than in cords from patients with non-ALS disorders. We further investigated the relationship between the level of BTBD10 immunoreactivity and the morphology of the Golgi apparatus (GA) and the presence of phosphorylated TAR-DNA-binding protein 43 (pTDP-43). Mirror sections of spinal cords from five sporadic ALS cases were immunostained with antibodies against BTBD10 and trans-Golgi-network (TGN)-46 or pTDP-43. Whereas 89.7-96.5% of the neurons with normal BTBD10 immunoreactivity showed normal GA morphology and no pTDP-43 cytoplasmic aggregates, 86.2-94.3% of the neurons with reduced BTBD10 expression showed GA fragmentation and abnormal pTDP-43 aggregates. These findings suggest that reduced BTBD10 expression is closely linked to the pathogenesis of sporadic ALS.

Furuta N; Makioka K; Fujita Y; Ikeda M; Takatama M; Matsuoka M; Okamoto K

2013-08-01

 
 
 
 
21

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

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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 ...

Xu Ya-Fei; Zhang Yong-Jie; Lin Wen-Lang; Cao Xiangkun; Stetler Caroline; Dickson Dennis W; Lewis Jada; Petrucelli Leonard

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Molecular Neuropathology of TDP-43 Proteinopathies  

Directory of Open Access Journals (Sweden)

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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Chronic cerebral ischemia induces redistribution and abnormal phosphorylation of transactivation-responsive DNA-binding protein-43 in mice.  

UK PubMed Central (United Kingdom)

Transactivation-responsive DNA-binding protein 43 (TDP-43) is closely involved in the pathogenesis of frontotemporal lobar degeneration. The native form, but not phosphorylated form, of TDP-43 has been reported to redistribute during acute neuronal injuries. Here, we examined whether the expression of phosphorylated TDP-43 was altered following chronic neuronal injury. C57BL/6 mice were subjected to sham operation or bilateral common carotid artery stenosis (BCAS) using microcoils, and changes in proteolytic cleavage, phosphorylation, and subcellular redistribution of TDP-43 were examined by immunoblotting and immunohistochemistry. We also monitored the expression of importin ?, which is involved in the transport of TDP-43. Immunoblotting showed an increase in phosphorylated TDP-43 in the nuclear fraction after BCAS using microcoils. Moreover, immunoreactivity toward phosphorylated TDP-43 was observed in the neuronal cytoplasm in the cerebral cortex and hippocampus, and importin ? levels decreased after the operation. Immunoreactivity toward phosphorylated TDP-43 was partly colocalized with immunoreactivity toward caspase 3 in the neuronal cytoplasm. These results suggested that chronic cerebral ischemia induced redistribution and abnormal phosphorylation of TDP-43, which may be triggered by downregulation of importin ? and may partly result in neuronal death.

Shindo A; Yata K; Sasaki R; Tomimoto H

2013-10-01

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Chronic cerebral ischemia induces redistribution and abnormal phosphorylation of transactivation-responsive DNA-binding protein-43 in mice.  

Science.gov (United States)

Transactivation-responsive DNA-binding protein 43 (TDP-43) is closely involved in the pathogenesis of frontotemporal lobar degeneration. The native form, but not phosphorylated form, of TDP-43 has been reported to redistribute during acute neuronal injuries. Here, we examined whether the expression of phosphorylated TDP-43 was altered following chronic neuronal injury. C57BL/6 mice were subjected to sham operation or bilateral common carotid artery stenosis (BCAS) using microcoils, and changes in proteolytic cleavage, phosphorylation, and subcellular redistribution of TDP-43 were examined by immunoblotting and immunohistochemistry. We also monitored the expression of importin ?, which is involved in the transport of TDP-43. Immunoblotting showed an increase in phosphorylated TDP-43 in the nuclear fraction after BCAS using microcoils. Moreover, immunoreactivity toward phosphorylated TDP-43 was observed in the neuronal cytoplasm in the cerebral cortex and hippocampus, and importin ? levels decreased after the operation. Immunoreactivity toward phosphorylated TDP-43 was partly colocalized with immunoreactivity toward caspase 3 in the neuronal cytoplasm. These results suggested that chronic cerebral ischemia induced redistribution and abnormal phosphorylation of TDP-43, which may be triggered by downregulation of importin ? and may partly result in neuronal death. PMID:23954745

Shindo, Akihiro; Yata, Kenichiro; Sasaki, Ryogen; Tomimoto, Hidekazu

2013-08-14

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Corticobasal degeneration with olivopontocerebellar atrophy and TDP-43 pathology: an unusual clinicopathologic variant of CBD.  

Science.gov (United States)

Corticobasal degeneration (CBD) is a disorder affecting cognition and movement due to a progressive neurodegeneration associated with distinctive neuropathologic features, including abnormal phosphorylated tau protein in neurons and glia in cortex, basal ganglia, diencephalon, and brainstem, as well as ballooned neurons and astrocytic plaques. We identified three cases of CBD with olivopontocerebellar atrophy (CBD-OPCA) that did not have ?-synuclein-positive glial cytoplasmic inclusions of multiple system atrophy (MSA). Two patients had clinical features suggestive of progressive supranuclear palsy (PSP), and the third case had cerebellar ataxia thought to be due to idiopathic OPCA. Neuropathologic features of CBD-OPCA are compared to typical CBD, as well as MSA and PSP. CBD-OPCA and MSA had marked neuronal loss in pontine nuclei, inferior olivary nucleus, and Purkinje cell layer. Neuronal loss and grumose degeneration in the cerebellar dentate nucleus were comparable in CBD-OPCA and PSP. Image analysis of tau pathology showed greater infratentorial tau burden, especially in pontine base, in CBD-OPCA compared with typical CBD. In addition, CBD-OPCA had TDP-43 immunoreactive neuronal and glial cytoplasmic inclusions and threads throughout the basal ganglia and in olivopontocerebellar system. CBD-OPCA met neuropathologic research diagnostic criteria for CBD and shared tau biochemical characteristics with typical CBD. These results suggest that CBD-OPCA is a distinct clinicopathologic variant of CBD with olivopontocerebellar TDP-43 pathology. PMID:23371366

Kouri, Naomi; Oshima, Kenichi; Takahashi, Makio; Murray, Melissa E; Ahmed, Zeshan; Parisi, Joseph E; Yen, Shu-Hui C; Dickson, Dennis W

2013-01-31

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Corticobasal degeneration with olivopontocerebellar atrophy and TDP-43 pathology: an unusual clinicopathologic variant of CBD.  

UK PubMed Central (United Kingdom)

Corticobasal degeneration (CBD) is a disorder affecting cognition and movement due to a progressive neurodegeneration associated with distinctive neuropathologic features, including abnormal phosphorylated tau protein in neurons and glia in cortex, basal ganglia, diencephalon, and brainstem, as well as ballooned neurons and astrocytic plaques. We identified three cases of CBD with olivopontocerebellar atrophy (CBD-OPCA) that did not have ?-synuclein-positive glial cytoplasmic inclusions of multiple system atrophy (MSA). Two patients had clinical features suggestive of progressive supranuclear palsy (PSP), and the third case had cerebellar ataxia thought to be due to idiopathic OPCA. Neuropathologic features of CBD-OPCA are compared to typical CBD, as well as MSA and PSP. CBD-OPCA and MSA had marked neuronal loss in pontine nuclei, inferior olivary nucleus, and Purkinje cell layer. Neuronal loss and grumose degeneration in the cerebellar dentate nucleus were comparable in CBD-OPCA and PSP. Image analysis of tau pathology showed greater infratentorial tau burden, especially in pontine base, in CBD-OPCA compared with typical CBD. In addition, CBD-OPCA had TDP-43 immunoreactive neuronal and glial cytoplasmic inclusions and threads throughout the basal ganglia and in olivopontocerebellar system. CBD-OPCA met neuropathologic research diagnostic criteria for CBD and shared tau biochemical characteristics with typical CBD. These results suggest that CBD-OPCA is a distinct clinicopathologic variant of CBD with olivopontocerebellar TDP-43 pathology.

Kouri N; Oshima K; Takahashi M; Murray ME; Ahmed Z; Parisi JE; Yen SH; Dickson DW

2013-05-01

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A 43-kDa TDP-43 species is present in aggregates associated with frontotemporal lobar degeneration.  

UK PubMed Central (United Kingdom)

The transactive response DNA-binding protein (TDP-43) is a major component of the abnormal intracellular inclusions that occur in two common neurodegenerative diseases of humans: (1) a subtype of frontotemporal lobar degeneration and (2) amyotrophic lateral sclerosis. Genetics, experiments in cultured cells and animals, and analogy with other neurodegenerative diseases indicate that the process of TDP-43 aggregation is fundamental to the pathogenesis of these 2 diseases, but the process by which this aggregation occurs is not understood. Biochemical fractionation has revealed truncated, phosphorylated and ubiquitinated forms of TDP-43 in a detergent-insoluble fraction from diseased CNS tissue, while these forms are absent from controls. However, a large amount of the normally predominant 43-kDa form of TDP-43 is present in the detergent-insoluble fraction even from control brains, so it has not been possible to determine if this form of TDP-43 is part of pathological aggregates in frontotemporal lobe degeneration. We used semi-denaturing detergent-agarose gel electrophoresis to isolate high molecular weight aggregates containing TDP-43 that are present in the cerebral cortex of individuals with frontotemporal lobar degeneration but not that of controls. These aggregates include the same covalently modified forms of TDP-43 seen in detergent-insoluble extracts. In addition, aggregates include a 43-kDa TDP-43 species. This aggregated 43-kDa form of TDP-43 is absent or present only at low levels in controls. The presence of 43-kDa TDP-43 in aggregates raises the possibility that covalent modification is not a primary step in the pathogenic aggregation of TDP-43 associated with frontotemporal lobar degeneration and amyotrophic lateral sclerosis.

Bosque PJ; Boyer PJ; Mishra P

2013-01-01

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TDP-43 deposition in prospectively followed, cognitively normal elderly individuals: correlation with argyrophilic grains but not other concomitant pathologies.  

Science.gov (United States)

TAR DNA-binding protein 43 (TDP-43) has been heavily researched in recent years due to its involvement in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration. Several studies have also sought to investigate the frequency of TDP-43 deposition in other neurodegenerative diseases such as Alzheimer's and Parkinson's diseases, but there has been relatively little work focused on the prevalence, distribution and histopathological associations of abnormal TDP-43 deposits in the brains of cognitively normal elderly subjects. We screened thick, free-floating coronal sections of mesial temporal lobe from 110 prospectively followed and autopsied cognitively normal subjects (age range 71-100 years) using an immunohistochemical method for phosphorylated TDP-43. We found a 36.4 % prevalence of pathologic TDP-43, mostly in the form of neurites while perikaryal cytoplasmic neuronal inclusions were uncommon and intranuclear inclusions were rare. With respect to other concomitant pathologies commonly found in elderly individuals, cases with TDP-43 had a greater prevalence of argyrophilic grains (ARG) (40 vs. 18.6 %) and overall ARG density (moderate vs. sparse). There were no additional associations with other concomitant pathologies, including cerebral white matter rarefaction, incidental Lewy bodies, neurofibrillary tangles or amyloid plaques. These results indicate deposition of TDP-43 occurs in a substantial subset of cognitively normal elderly subjects and is more common in those with ARG, supporting some previous studies linking pathological TDP-43 deposition with ARG and other pathological tau protein deposits. PMID:23604587

Arnold, Stacy J; Dugger, Brittany N; Beach, Thomas G

2013-04-20

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CDC7 inhibition blocks pathological TDP-43 phosphorylation and neurodegeneration.  

UK PubMed Central (United Kingdom)

OBJECTIVE: Kinase hyperactivity occurs in both neurodegenerative disease and cancer. Lesions containing hyperphosphorylated aggregated TDP-43 characterize amyotrophic lateral sclerosis and frontotemporal lobar degeneration with TDP-43 inclusions. Dual phosphorylation of TDP-43 at serines 409/410 (S409/410) drives neurotoxicity in disease models; therefore, TDP-43-specific kinases are candidate targets for intervention. METHODS: To find therapeutic targets for the prevention of TDP-43 phosphorylation, we assembled and screened a comprehensive RNA interference library targeting kinases in TDP-43 transgenic Caenorhabditis elegans. RESULTS: We show CDC7 robustly phosphorylates TDP-43 at pathological residues S409/410 in C. elegans, in vitro, and in human cell culture. In frontotemporal lobar degeneration (FTLD)-TDP cases, CDC7 immunostaining overlaps with the phospho-TDP-43 pathology found in frontal cortex. Furthermore, PHA767491, a small molecule inhibitor of CDC7, reduces TDP-43 phosphorylation and prevents TDP-43-dependent neurodegeneration in TDP-43-transgenic animals. INTERPRETATION: Taken together, these data support CDC7 as a novel therapeutic target for TDP-43 proteinopathies, including FTLD-TDP and amyotrophic lateral sclerosis. Ann Neurol 2013;74:39-52.

Liachko NF; McMillan PJ; Guthrie CR; Bird TD; Leverenz JB; Kraemer BC

2013-07-01

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Identification of genetic modifiers of TDP-43 neurotoxicity in Drosophila.  

UK PubMed Central (United Kingdom)

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 sufficient to induce neurotoxicity, paralysis, and early death. Using microarrays, we have examined gene expression patterns that accompany TDP-43-induced neurotoxicity in the fly system. Constitutive expression of TDP-43 in the Drosophila compound eye elicited widespread gene expression changes, with strong upregulation of cell cycle regulatory genes and genes functioning in the Notch intercellular communication pathway. Inducible expression of TDP-43 specifically in neurons elicited significant expression differences in a more restricted set of genes. Genes that were upregulated in both paradigms included SpindleB and the Notch target Hey, which appeared to be a direct TDP-43 target. Mutations that diminished activity of Notch or disrupted the function of downstream Notch target genes extended the lifespan of TDP-43 transgenic flies, suggesting that Notch activation was deleterious in this model. Finally, we showed that mutation of the nucleoporin Nup50 increased the lifespan of TDP-43 transgenic flies, suggesting that nuclear events contribute to TDP-43-dependent neurotoxicity. The combined findings identified pathways whose deregulation might contribute to TDP-43-induced neurotoxicity in Drosophila.

Zhan L; Hanson KA; Kim SH; Tare A; Tibbetts RS

2013-01-01

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

UK PubMed Central (United Kingdom)

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.

Diaper DC; Adachi Y; Lazarou L; Greenstein M; Simoes FA; Di Domenico A; Solomon DA; Lowe S; Alsubaie R; Cheng D; Buckley S; Humphrey DM; Shaw CE; Hirth F

2013-10-01

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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

2013-05-31

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Inhibition of TDP-43 aggregation by nucleic acid binding.  

UK PubMed Central (United Kingdom)

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 on the in vitro protein expression system to evaluate the solubility change of TDP-43 in response to factors such as nucleotide binding and temperature. Our results suggest that the solubility of TDP-43 is largely influenced by its cognate single-strand DNA (ssDNA) or RNA (ssRNA) rather than hnRNP, which is known to associate with TDP-43 C-terminus. The direct interaction between the refolded TDP-43, purified from E.coli, and ssDNA were further characterized by Circular Dichroism (CD) as well as turbidity and filter binding assay. In addition, ssDNA or ssRNA failed to prevent the aggregation of the F147L/F149L double mutant or truncated TDP-43 (TDP208-414). Consistently, these two mutants form aggregates, in contrast with the wild-type TDP-43, when expressed in Neuro2a cells. Our results demonstrate an intimate relationship between the solubility of TDP-43 and its DNA or RNA binding affinity, which may shed light on the role of TDP-43 in ALS and FTLD.

Huang YC; Lin KF; He RY; Tu PH; Koubek J; Hsu YC; Huang JJ

2013-01-01

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Laminar distribution of the pathological changes in sporadic frontotemporal lobar degeneration with transactive response (TAR) DNA-binding protein of 43 kDa (TDP-43) proteinopathy: a quantitative study using polynomial curve fitting.  

UK PubMed Central (United Kingdom)

AIMS: Previous data suggest heterogeneity in laminar distribution of the pathology in the molecular disorder frontotemporal lobar degeneration (FTLD) with transactive response (TAR) DNA-binding protein of 43 kDa (TDP-43) proteinopathy (FTLD-TDP). To study this heterogeneity, we quantified the changes in density across the cortical laminae of neuronal cytoplasmic inclusions, glial inclusions, neuronal intranuclear inclusions, dystrophic neurites, surviving neurones, abnormally enlarged neurones, and vacuoles in regions of the frontal and temporal lobe. METHODS: Changes in density of histological features across cortical gyri were studied in 10 sporadic cases of FTLD-TDP using quantitative methods and polynomial curve fitting. RESULTS: Our data suggest that laminar neuropathology in sporadic FTLD-TDP is highly variable. Most commonly, neuronal cytoplasmic inclusions, dystrophic neurites and vacuolation were abundant in the upper laminae and glial inclusions, neuronal intranuclear inclusions, abnormally enlarged neurones, and glial cell nuclei in the lower laminae. TDP-43-immunoreactive inclusions affected more of the cortical profile in longer duration cases; their distribution varied with disease subtype, but was unrelated to Braak tangle score. Different TDP-43-immunoreactive inclusions were not spatially correlated. CONCLUSIONS: Laminar distribution of pathological features in 10 sporadic cases of FTLD-TDP is heterogeneous and may be accounted for, in part, by disease subtype and disease duration. In addition, the feedforward and feedback cortico-cortical connections may be compromised in FTLD-TDP.

Armstrong RA; Hamilton RL; Mackenzie IR; Hedreen J; Cairns NJ

2013-06-01

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The truncated C-terminal RNA recognition motif of TDP-43 protein plays a key role in forming proteinaceous aggregates.  

UK PubMed Central (United Kingdom)

TDP-43 is the major pathological protein identified in the cellular inclusions in amyotrophic lateral sclerosis and frontotemporal lobar degeneration. The pathogenic forms of TDP-43 are processed C-terminal fragments containing a truncated RNA-recognition motif (RRM2) and a glycine-rich region. Although extensive studies have focused on this protein, it remains unclear how the dimeric full-length TDP-43 is folded and assembled and how the processed C-terminal fragments are misfolded and aggregated. Here, using size-exclusion chromatography, pulldown assays, and small angle x-ray scattering, we show that the C-terminal-deleted TDP-43 without the glycine-rich tail is sufficient to form a head-to-head homodimer primarily via its N-terminal domain. The truncated RRM2, as well as two ?-strands within the RRM2, form fibrils in vitro with a similar amyloid-negative staining property to those of TDP-43 pathogenic fibrils in diseases. In addition to the glycine-rich region, the truncated RRM2, but not the intact RRM2, plays a key role in forming cytoplasmic inclusions in neuronal cells. Our data thus suggest that the process that disrupts the dimeric structure, such as the proteolytic cleavage of TDP-43 within the RRM2 that removes the N-terminal dimerization domain, may produce unassembled truncated RRM2 fragments with abnormally exposed ?-strands, which can oligomerize into high-order inclusions.

Wang YT; Kuo PH; Chiang CH; Liang JR; Chen YR; Wang S; Shen JC; Yuan HS

2013-03-01

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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

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ALS-linked TDP-43 mutations produce aberrant RNA splicing and adult-onset motor neuron disease without aggregation or loss of nuclear TDP-43  

Science.gov (United States)

Transactivating response region DNA binding protein (TDP-43) is the major protein component of ubiquitinated inclusions found in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) with ubiquitinated inclusions. Two ALS-causing mutants (TDP-43Q331K and TDP-43M337V), but not wild-type human TDP-43, are shown here to provoke age-dependent, mutant-dependent, progressive motor axon degeneration and motor neuron death when expressed in mice at levels and in a cell type-selective pattern similar to endogenous TDP-43. Mutant TDP-43-dependent degeneration of lower motor neurons occurs without: (i) loss of TDP-43 from the corresponding nuclei, (ii) accumulation of TDP-43 aggregates, and (iii) accumulation of insoluble TDP-43. Computational analysis using splicing-sensitive microarrays demonstrates alterations of endogenous TDP-43–dependent alternative splicing events conferred by both human wild-type and mutant TDP-43Q331K, but with high levels of mutant TDP-43 preferentially enhancing exon exclusion of some target pre-mRNAs affecting genes involved in neurological transmission and function. Comparison with splicing alterations following TDP-43 depletion demonstrates that TDP-43Q331K enhances normal TDP-43 splicing function for some RNA targets but loss-of-function for others. Thus, adult-onset motor neuron disease does not require aggregation or loss of nuclear TDP-43, with ALS-linked mutants producing loss and gain of splicing function of selected RNA targets at an early disease stage.

Arnold, Eveline S.; Ling, Shuo-Chien; Huelga, Stephanie C.; Lagier-Tourenne, Clotilde; Polymenidou, Magdalini; Ditsworth, Dara; Kordasiewicz, Holly B.; McAlonis-Downes, Melissa; Platoshyn, Oleksandr; Parone, Philippe A.; Da Cruz, Sandrine; Clutario, Kevin M.; Swing, Debbie; Tessarollo, Lino; Marsala, Martin; Shaw, Christopher E.; Yeo, Gene W.; Cleveland, Don W.

2013-01-01

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ALS-linked TDP-43 mutations produce aberrant RNA splicing and adult-onset motor neuron disease without aggregation or loss of nuclear TDP-43.  

UK PubMed Central (United Kingdom)

Transactivating response region DNA binding protein (TDP-43) is the major protein component of ubiquitinated inclusions found in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) with ubiquitinated inclusions. Two ALS-causing mutants (TDP-43(Q331K) and TDP-43(M337V)), but not wild-type human TDP-43, are shown here to provoke age-dependent, mutant-dependent, progressive motor axon degeneration and motor neuron death when expressed in mice at levels and in a cell type-selective pattern similar to endogenous TDP-43. Mutant TDP-43-dependent degeneration of lower motor neurons occurs without: (i) loss of TDP-43 from the corresponding nuclei, (ii) accumulation of TDP-43 aggregates, and (iii) accumulation of insoluble TDP-43. Computational analysis using splicing-sensitive microarrays demonstrates alterations of endogenous TDP-43-dependent alternative splicing events conferred by both human wild-type and mutant TDP-43(Q331K), but with high levels of mutant TDP-43 preferentially enhancing exon exclusion of some target pre-mRNAs affecting genes involved in neurological transmission and function. Comparison with splicing alterations following TDP-43 depletion demonstrates that TDP-43(Q331K) enhances normal TDP-43 splicing function for some RNA targets but loss-of-function for others. Thus, adult-onset motor neuron disease does not require aggregation or loss of nuclear TDP-43, with ALS-linked mutants producing loss and gain of splicing function of selected RNA targets at an early disease stage.

Arnold ES; Ling SC; Huelga SC; Lagier-Tourenne C; Polymenidou M; Ditsworth D; Kordasiewicz HB; McAlonis-Downes M; Platoshyn O; Parone PA; Da Cruz S; Clutario KM; Swing D; Tessarollo L; Marsala M; Shaw CE; Yeo GW; Cleveland DW

2013-02-01

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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

2012-01-01

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TDP-43 toxicity and the usefulness of junk.  

Science.gov (United States)

A new study shows that loss of the lariat debranching enzyme Dbr1 suppresses TDP-43 toxicity. The accumulated intronic lariat RNAs, which are normally degraded after splicing, likely act as decoys to sequester TDP-43 away from binding to and disrupting functions of other RNAs. PMID:23192178

Sun, Shuying; Cleveland, Don W

2012-12-01

 
 
 
 
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TDP-43 toxicity and the usefulness of junk.  

UK PubMed Central (United Kingdom)

A new study shows that loss of the lariat debranching enzyme Dbr1 suppresses TDP-43 toxicity. The accumulated intronic lariat RNAs, which are normally degraded after splicing, likely act as decoys to sequester TDP-43 away from binding to and disrupting functions of other RNAs.

Sun S; Cleveland DW

2012-12-01

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Kinase Inhibitor Screening Identifies Cyclin-Dependent Kinases and Glycogen Synthase Kinase 3 as Potential Modulators of TDP-43 Cytosolic Accumulation during Cell Stress.  

UK PubMed Central (United Kingdom)

Abnormal processing of TAR DNA binding protein 43 (TDP-43) has been identified as a major factor in neuronal degeneration during amyotrophic lateral sclerosis (ALS) or frontotemporal lobar degeneration (FTLD). It is unclear how changes to TDP-43, including nuclear to cytosolic translocation and subsequent accumulation, are controlled in these diseases. TDP-43 is a member of the heterogeneous ribonucleoprotein (hnRNP) RNA binding protein family and is known to associate with cytosolic RNA stress granule proteins in ALS and FTLD. hnRNP trafficking and accumulation is controlled by the action of specific kinases including members of the mitogen-activated protein kinase (MAPK) pathway. However, little is known about how kinase pathways control TDP-43 movement and accumulation. In this study, we used an in vitro model of TDP-43-positve stress granule formation to screen for the effect of kinase inhibitors on TDP-43 accumulation. We found that while a number of kinase inhibitors, particularly of the MAPK pathways modulated both TDP-43 and the global stress granule marker, human antigen R (HuR), multiple inhibitors were more specific to TDP-43 accumulation, including inhibitors of cyclin-dependent kinases (CDKs) and glycogen synthase kinase 3 (GSK3). Close correlation was observed between effects of these inhibitors on TDP-43, hnRNP K and TIAR, but often with different effects on HuR accumulation. This may indicate a potential interaction between TDP-43, hnRNP K and TIAR. CDK inhibitors were also found to reverse pre-formed TDP-43-positive stress granules and both CDK and GSK3 inhibitors abrogated the accumulation of C-terminal TDP-43 (219-414) in transfected cells. Further studies are required to confirm the specific kinases involved and whether their action is through phosphorylation of the TDP-43 binding partner hnRNP K. This knowledge provides a valuable insight into the mechanisms controlling abnormal cytoplasmic TDP-43 accumulation and may herald new opportunities for kinase modulation-based therapeutic intervention in ALS and FTLD.

Moujalled D; James JL; Parker SJ; Lidgerwood GE; Duncan C; Meyerowitz J; Nonaka T; Hasegawa M; Kanninen KM; Grubman A; Liddell JR; Crouch PJ; White AR

2013-01-01

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Expression of TDP-43 C-terminal fragments in vitro recapitulates pathological features of TDP-43 proteinopathies  

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The disease protein in frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U) and amyotrophic lateral sclerosis (ALS) was identified recently as the TDP-43 (TAR DNA-binding protein 43), thereby providing a molecular link between these two disorders. In FTLD-U and ALS, TDP-43 i...

Igaz, L M; Kwong, L K; Chen-Plotkin, A; Winton, M J; Unger, T L; Xu, Y; Neumann, M; Trojanowski, J Q; Lee, V M Y

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Characterizing TDP-43 interaction with its RNA targets.  

UK PubMed Central (United Kingdom)

One of the most important functional features of nuclear factor TDP-43 is its ability to bind UG-repeats with high efficiency. Several cross-linking and immunoprecipitation (CLIP) and RNA immunoprecipitation-sequencing (RIP-seq) analyses have indicated that TDP-43 in vivo can also specifically bind loosely conserved UG/GU-rich repeats interspersed by other nucleotides. These sequences are predominantly localized within long introns and in the 3'UTR of various genes. Most importantly, some of these sequences have been found to exist in the 3'UTR region of TDP-43 itself. In the TDP-43 3'UTR context, the presence of these UG-like sequences is essential for TDP-43 to autoregulate its own levels through a negative feedback loop. In this work, we have compared the binding of TDP-43 with these types of sequences as opposed to perfect UG-stretches. We show that the binding affinity to the UG-like sequences has a dissociation constant (Kd) of ?110 nM compared with a Kd of 8 nM for straight UGs, and have mapped the region of contact between protein and RNA. In addition, our results indicate that the local concentration of UG dinucleotides in the CLIP sequences is one of the major factors influencing the interaction of these RNA sequences with TDP-43.

Bhardwaj A; Myers MP; Buratti E; Baralle FE

2013-05-01

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Premature death of TDP-43 (A315T) transgenic mice due to gastrointestinal complications prior to development of full neurological symptoms of amyotrophic lateral sclerosis.  

UK PubMed Central (United Kingdom)

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). Transgenic mouse lines overexpressing wild-type or mutant TDP-43 exhibit ALS-like symptom, motor abnormalities and early paralysis followed by death. Reports on lifespan and phenotypic behaviour in Prp-TDP-43 (A315T) vary, and these animals are not fully characterized. Although it has been proposed that the approximate 20% loss of motor neurons at end stage is responsible for the severe weakness and death in TDP-43 mice, this degree of neurologic damage appears insufficient to cause death. Hence we studied these mice to further characterize and determine the reason for the death. Our characterization of TDP-43 transgenic mice showed that these mice develop ALS-like symptoms that later become compounded by gastrointestinal (GI) complications that resulted in death. This is the first report of a set of pathological evidence in the GI track that is strong indicator for the cause of death of Prp-hTDP-43 (A315T) transgenic mice.

Esmaeili MA; Panahi M; Yadav S; Hennings L; Kiaei M

2013-02-01

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Asymmetric TDP-43 distribution in primary progressive aphasia with progranulin mutation.  

UK PubMed Central (United Kingdom)

OBJECTIVE: Primary progressive aphasia (PPA) results from an asymmetric degeneration of the language dominant (usually left) hemisphere and can be associated with the pathology of Alzheimer disease (AD) or frontotemporal lobar degeneration (FTLD). This study aimed to investigate whether the anatomic distribution of TDP-43 inclusions displayed a corresponding leftward asymmetry in a patient with PPA with a mutation in the progranulin gene and FTLD pathology. METHODS: Brain tissue from a 65-year-old patient with PPA and progranulin mutation was analyzed using immunohistochemical methods for TDP-43. Analysis was performed in the superior temporal gyrus, inferior temporal gyrus, inferior parietal lobule, orbitofrontal cortex, entorhinal cortex, and dentate gyrus. Neuronal intranuclear inclusions, neuronal cytoplasmic inclusions, and dystrophic neurites were quantified using modified stereologic analysis. Analysis of variance was used to determine significant effects. RESULTS: All 3 types of inclusions predominated on the left side of analyzed cortical regions. They were also more frequent in language areas than in memory-related areas. CONCLUSION: These results demonstrate a phenotypically concordant distribution of abnormal TDP-43 inclusions in primary progressive aphasia (PPA). This contrasts with PPA cases with Alzheimer pathology where no consistent leftward asymmetry of neurofibrillary degeneration or amyloid deposition has been demonstrated despite the leftward asymmetry of the atrophy, and where neurofibrillary tangles show a greater density in memory than language areas despite the predominantly aphasic phenotype. This case suggests that the TDP-43 inclusions in PPA-frontotemporal lobar degeneration are more tightly linked to neuronal death and dysfunction than neurofibrillary and amyloid deposits in PPA-Alzheimer disease.

Gliebus G; Bigio EH; Gasho K; Mishra M; Caplan D; Mesulam MM; Geula C

2010-05-01

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Overexpression of ALS-associated p.M337V human TDP-43 in mice worsens disease features compared to wild-type human TDP-43 mice.  

UK PubMed Central (United Kingdom)

Mutations in TAR DNA-binding protein 43 (TDP-43) are associated with familial forms of amyotrophic lateral sclerosis (ALS), while wild-type TDP-43 is a pathological hallmark of patients with sporadic ALS and frontotemporal lobar degeneration (FTLD). Various in vitro and in vivo studies have also demonstrated toxicity of both mutant and wild-type TDP-43 to neuronal cells. To study the potential additional toxicity incurred by mutant TDP-43 in vivo, we generated mutant human TDP-43 (p.M337V) transgenic mouse lines driven by the Thy-1.2 promoter (Mt-TAR) and compared them in the same experimental setting to the disease phenotype observed in wild-type TDP-43 transgenic lines (Wt-TAR) expressing comparable TDP-43 levels. Overexpression of mutant TDP-43 leads to a worsened dose-dependent disease phenotype in terms of motor dysfunction, neurodegeneration, gliosis, and development of ubiquitin and phosphorylated TDP-43 pathology. Furthermore, we show that cellular aggregate formation or accumulation of TDP-43 C-terminal fragments (CTFs) are not primarily responsible for development of the observed disease phenotype in both mutant and wild-type TDP-43 mice.

Janssens J; Wils H; Kleinberger G; Joris G; Cuijt I; Ceuterick-de Groote C; Van Broeckhoven C; Kumar-Singh S

2013-08-01

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TDP-43 and FUS RNA-binding proteins bind distinct sets of cytoplasmic messenger RNAs and differently regulate their post-transcriptional fate in motoneuron-like cells.  

UK PubMed Central (United Kingdom)

The RNA-binding proteins TDP-43 and FUS form abnormal cytoplasmic aggregates in affected tissues of patients with amyotrophic lateral sclerosis and frontotemporal lobar dementia. TDP-43 and FUS localize mainly in the nucleus where they regulate pre-mRNA splicing, but they are also involved in mRNA transport, stability, and translation. To better investigate their cytoplasmic activities, we applied an RNA immunoprecipitation and chip analysis to define the mRNAs associated to TDP-43 and FUS in the cytoplasmic ribonucleoprotein complexes from motoneuronal NSC-34 cells. We found that they bind different sets of mRNAs although converging on common cellular pathways. Bioinformatics analyses identified the (UG)(n) consensus motif in 80% of 3'-UTR sequences of TDP-43 targets, whereas for FUS the binding motif was less evident. By in vitro assays we validated binding to selected target 3'-UTRs, including Vegfa and Grn for TDP-43, and Vps54, Nvl, and Taf15 for FUS. We showed that TDP-43 has a destabilizing activity on Vegfa and Grn mRNAs and may ultimately affect progranulin protein content, whereas FUS does not affect mRNA stability/translation of its targets. We also demonstrated that three different point mutations in TDP-43 did not change the binding affinity for Vegfa and Grn mRNAs or their protein level. Our data indicate that TDP-43 and FUS recognize distinct sets of mRNAs and differently regulate their fate in the cytoplasm of motoneuron-like cells, therefore suggesting complementary roles in neuronal RNA metabolism and neurodegeneration.

Colombrita C; Onesto E; Megiorni F; Pizzuti A; Baralle FE; Buratti E; Silani V; Ratti A

2012-05-01

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TDP-43 and FUS RNA-binding Proteins Bind Distinct Sets of Cytoplasmic Messenger RNAs and Differently Regulate Their Post-transcriptional Fate in Motoneuron-like Cells*  

Science.gov (United States)

The RNA-binding proteins TDP-43 and FUS form abnormal cytoplasmic aggregates in affected tissues of patients with amyotrophic lateral sclerosis and frontotemporal lobar dementia. TDP-43 and FUS localize mainly in the nucleus where they regulate pre-mRNA splicing, but they are also involved in mRNA transport, stability, and translation. To better investigate their cytoplasmic activities, we applied an RNA immunoprecipitation and chip analysis to define the mRNAs associated to TDP-43 and FUS in the cytoplasmic ribonucleoprotein complexes from motoneuronal NSC-34 cells. We found that they bind different sets of mRNAs although converging on common cellular pathways. Bioinformatics analyses identified the (UG)n consensus motif in 80% of 3?-UTR sequences of TDP-43 targets, whereas for FUS the binding motif was less evident. By in vitro assays we validated binding to selected target 3?-UTRs, including Vegfa and Grn for TDP-43, and Vps54, Nvl, and Taf15 for FUS. We showed that TDP-43 has a destabilizing activity on Vegfa and Grn mRNAs and may ultimately affect progranulin protein content, whereas FUS does not affect mRNA stability/translation of its targets. We also demonstrated that three different point mutations in TDP-43 did not change the binding affinity for Vegfa and Grn mRNAs or their protein level. Our data indicate that TDP-43 and FUS recognize distinct sets of mRNAs and differently regulate their fate in the cytoplasm of motoneuron-like cells, therefore suggesting complementary roles in neuronal RNA metabolism and neurodegeneration.

Colombrita, Claudia; Onesto, Elisa; Megiorni, Francesca; Pizzuti, Antonio; Baralle, Francisco E.; Buratti, Emanuele; Silani, Vincenzo; Ratti, Antonia

2012-01-01

50

TDP-43 and FUS RNA-binding proteins bind distinct sets of cytoplasmic messenger RNAs and differently regulate their post-transcriptional fate in motoneuron-like cells.  

Science.gov (United States)

The RNA-binding proteins TDP-43 and FUS form abnormal cytoplasmic aggregates in affected tissues of patients with amyotrophic lateral sclerosis and frontotemporal lobar dementia. TDP-43 and FUS localize mainly in the nucleus where they regulate pre-mRNA splicing, but they are also involved in mRNA transport, stability, and translation. To better investigate their cytoplasmic activities, we applied an RNA immunoprecipitation and chip analysis to define the mRNAs associated to TDP-43 and FUS in the cytoplasmic ribonucleoprotein complexes from motoneuronal NSC-34 cells. We found that they bind different sets of mRNAs although converging on common cellular pathways. Bioinformatics analyses identified the (UG)(n) consensus motif in 80% of 3'-UTR sequences of TDP-43 targets, whereas for FUS the binding motif was less evident. By in vitro assays we validated binding to selected target 3'-UTRs, including Vegfa and Grn for TDP-43, and Vps54, Nvl, and Taf15 for FUS. We showed that TDP-43 has a destabilizing activity on Vegfa and Grn mRNAs and may ultimately affect progranulin protein content, whereas FUS does not affect mRNA stability/translation of its targets. We also demonstrated that three different point mutations in TDP-43 did not change the binding affinity for Vegfa and Grn mRNAs or their protein level. Our data indicate that TDP-43 and FUS recognize distinct sets of mRNAs and differently regulate their fate in the cytoplasm of motoneuron-like cells, therefore suggesting complementary roles in neuronal RNA metabolism and neurodegeneration. PMID:22427648

Colombrita, Claudia; Onesto, Elisa; Megiorni, Francesca; Pizzuti, Antonio; Baralle, Francisco E; Buratti, Emanuele; Silani, Vincenzo; Ratti, Antonia

2012-03-16

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Prion-like properties of pathological TDP-43 aggregates from diseased brains.  

UK PubMed Central (United Kingdom)

TDP-43 is the major component protein of ubiquitin-positive inclusions in brains of patients with frontotemporal lobar degeneration (FTLD-TDP) or amyotrophic lateral sclerosis (ALS). Here, we report the characterization of prion-like properties of aggregated TDP-43 prepared from diseased brains. When insoluble TDP-43 from ALS or FTLD-TDP brains was introduced as seeds into SH-SY5Y cells expressing TDP-43, phosphorylated and ubiquitinated TDP-43 was aggregated in a self-templating manner. Immunoblot analyses revealed that the C-terminal fragments of insoluble TDP-43 characteristic of each disease type acted as seeds, inducing seed-dependent aggregation of TDP-43 in these cells. The seeding ability of insoluble TDP-43 was unaffected by proteinase treatment but was abrogated by formic acid. One subtype of TDP-43 aggregate was resistant to boiling treatment. The insoluble fraction from cells harboring TDP-43 aggregates could also trigger intracellular TDP-43 aggregation. These results indicate that insoluble TDP-43 has prion-like properties that may play a role in the progression of TDP-43 proteinopathy.

Nonaka T; Masuda-Suzukake M; Arai T; Hasegawa Y; Akatsu H; Obi T; Yoshida M; Murayama S; Mann DM; Akiyama H; Hasegawa M

2013-07-01

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Characterization of ?-domains in C-terminal fragments of TDP-43 by scanning tunneling microscopy.  

UK PubMed Central (United Kingdom)

The TAR DNA-binding protein 43 (TDP-43) has been identified as a critical player in a range of neurodegenerative diseases, including frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). Recent discoveries demonstrate the important role of carboxyl-terminal fragments of TDP-43 in its proteinopathy. Herein, we report the characterization of ?-domains in the C-terminal fragments of TDP-43 using scanning tunneling microscopy (STM). Careful comparison of the wild-type TDP-43 (Wt) and the three mutant TDP-43 peptides: an ALS-related mutant peptide: phosphorylated A315T mutant TDP-43 (A315T(p)) and two model peptides: A315T mutant TDP-43 (A315T), A315E mutant TDP-43 (A315E) reveals that A315T(p) has a longer core region of the ?-domain than Wt. A315E possesses the longest core region of the ?-domain and A315T(p) mutant TDP-43 has the second longest core region of the ?-domain. The core regions of the ?-domains for A315T and Wt TDP-43 have the same length. This observation provides a supportive evidence of a higher tendency in beta-sheet formation of A315T(p) containing TDP-43 fragment, and structural mechanism for the higher cytotoxicity and accelerated fibril formation of the A315T(p) mutation-containing TDP-43 peptide as compared with Wt TDP-43.

Xu M; Zhu L; Liu J; Yang Y; Wu JY; Wang C

2013-01-01

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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 scle...

Zhang, Yong-Jie; Xu, Ya-Fei; Cook, Casey; Gendron, Tania F.; Roettges, Paul; Link, Christopher D.; Lin, Wen-Lang

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

Directory of Open Access Journals (Sweden)

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

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Identification of RNA bound to the TDP-43 ribonucleoprotein complex in the adult mouse brain.  

UK PubMed Central (United Kingdom)

Cytoplasmic inclusions containing TDP-43 are a pathological hallmark of several neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. TDP-43 is an RNA binding protein involved in gene regulation through control of RNA transcription, splicing and transport. However, the function of TDP-43 in the nervous system is largely unknown and its role in the pathogenesis of ALS is unclear. The aim of this study was to identify genes in the central nervous system that are regulated by TDP-43. RNA-immunoprecipitation with anti-TDP-43 antibody, followed by microarray analysis (RIP-chip), was used to isolate and identify RNA bound to TDP-43 protein from mouse brain. This analysis produced a list of 1839 potential TDP-43 gene targets, many of which overlap with previous studies and whose functions include RNA processing and synaptic function. Immunohistochemistry demonstrated that the TDP-43 protein could be found at the presynaptic membrane of axon terminals in the neuromuscular junction in mice. In conclusion, the finding that TDP-43 binds to RNA that codes for genes related to synaptic function, together with the localization of TDP-43 protein at axon terminals, suggests a role for TDP-43 in the transport of synaptic mRNAs into distal processes.

Narayanan RK; Mangelsdorf M; Panwar A; Butler TJ; Noakes PG; Wallace RH

2013-05-01

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Identification of RNA bound to the TDP-43 ribonucleoprotein complex in the adult mouse brain.  

Science.gov (United States)

Cytoplasmic inclusions containing TDP-43 are a pathological hallmark of several neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. TDP-43 is an RNA binding protein involved in gene regulation through control of RNA transcription, splicing and transport. However, the function of TDP-43 in the nervous system is largely unknown and its role in the pathogenesis of ALS is unclear. The aim of this study was to identify genes in the central nervous system that are regulated by TDP-43. RNA-immunoprecipitation with anti-TDP-43 antibody, followed by microarray analysis (RIP-chip), was used to isolate and identify RNA bound to TDP-43 protein from mouse brain. This analysis produced a list of 1839 potential TDP-43 gene targets, many of which overlap with previous studies and whose functions include RNA processing and synaptic function. Immunohistochemistry demonstrated that the TDP-43 protein could be found at the presynaptic membrane of axon terminals in the neuromuscular junction in mice. In conclusion, the finding that TDP-43 binds to RNA that codes for genes related to synaptic function, together with the localization of TDP-43 protein at axon terminals, suggests a role for TDP-43 in the transport of synaptic mRNAs into distal processes. PMID:23134510

Narayanan, Ramesh K; Mangelsdorf, Marie; Panwar, Ajay; Butler, Tim J; Noakes, Peter G; Wallace, Robyn H

2012-10-24

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

Directory of Open Access Journals (Sweden)

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; Ferris Jacob; Gao Fen-Biao

2009-01-01

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Ubiquilin-2 (UBQLN2) binds with high affinity to the C-terminal region of TDP-43 and modulates TDP-43 levels in H4 cells: characterization of inhibition by nucleic acids and 4-aminoquinolines.  

UK PubMed Central (United Kingdom)

Recently, it was reported that mutations in the ubiquitin-like protein ubiquilin-2 (UBQLN2) are associated with X-linked amyotrophic lateral sclerosis (ALS), and that both wild-type and mutant UBQLN2 can co-localize with aggregates of C-terminal fragments of TAR DNA binding protein (TDP-43). Here, we describe a high affinity interaction between UBQLN2 and TDP-43 and demonstrate that overexpression of both UBQLN2 and TDP-43 reduces levels of both exogenous and endogenous TDP-43 in human H4 cells. UBQLN2 bound with high affinity to both full length TDP-43 and a C-terminal TDP-43 fragment (261-414 aa) with KD values of 6.2nM and 8.7nM, respectively. Both DNA oligonucleotides and 4-aminoquinolines, which bind to TDP-43, also inhibited UBQLN2 binding to TDP-43 with similar rank order affinities compared to inhibition of oligonucleotide binding to TDP-43. Inhibitor characterization experiments demonstrated that the DNA oligonucleotides noncompetitively inhibited UBQLN2 binding to TDP-43, which is consistent with UBQLN2 binding to the C-terminal region of TDP-43. Interestingly, the 4-aminoquinolines were competitive inhibitors of UBQLN2 binding to TDP-43, suggesting that these compounds also bind to the C-terminal region of TDP-43. In support of the biochemical data, co-immunoprecipitation experiments demonstrated that both TDP-43 and UBQLN2 interact in human neuroglioma H4 cells. Finally, overexpression of UBQLN2 in the presence of overexpressed full length TDP-43 or C-terminal TDP-43 (170-414) dramatically lowered levels of both full length TDP-43 and C-terminal TDP-43 fragments (CTFs). Consequently, these data suggest that UBQLN2 enhances the clearance of TDP-43 and TDP-43 CTFs and therefore may play a role in the development of TDP-43 associated neurotoxicity.

Cassel JA; Reitz AB

2013-06-01

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Neuronal-specific overexpression of a mutant valosin-containing protein associated with IBMPFD promotes aberrant ubiquitin and TDP-43 accumulation and cognitive dysfunction in transgenic mice.  

UK PubMed Central (United Kingdom)

Mutations in valosin-containing protein (VCP) cause a rare, autosomal dominant disease called inclusion body myopathy associated with Paget disease of bone and frontotemporal dementia (IBMPFD). One-third of patients with IBMPFD develop frontotemporal dementia, characterized by an extensive neurodegeneration in the frontal and temporal lobes. Neuropathologic hallmarks include nuclear and cytosolic inclusions positive to ubiquitin and transactive response DNA-binding protein 43 (TDP-43) in neurons and glial activation in affected regions. However, the pathogenic mechanisms by which mutant VCP triggers neurodegeneration remain unknown. Herein, we generated a mouse model selectively overexpressing a human mutant VCP in neurons to study pathogenic mechanisms of mutant VCP-mediated neurodegeneration and cognitive impairment. The overexpression of VCPA232E mutation in forebrain regions produced significant progressive impairments of cognitive function, including deficits in spatial memory, object recognition, and fear conditioning. Although overexpressed or endogenous VCP did not seem to focally aggregate inside neurons, TDP-43 and ubiquitin accumulated with age in transgenic mouse brains. TDP-43 was also found to co-localize with stress granules in the cytosolic compartment. Together with the appearance of high-molecular-weight TDP-43 in cytosolic fractions, these findings demonstrate the mislocalization and accumulation of abnormal TDP-43 in the cytosol of transgenic mice, which likely lead to an increase in cellular stress and cognitive impairment. Taken together, these results highlight an important pathologic link between VCP and cognition.

Rodriguez-Ortiz CJ; Hoshino H; Cheng D; Liu-Yescevitz L; Blurton-Jones M; Wolozin B; LaFerla FM; Kitazawa M

2013-08-01

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Heteronemin, a marine sponge terpenoid, targets TDP-43, a key factor in several neurodegenerative disorders.  

UK PubMed Central (United Kingdom)

Trans-activation response DNA-binding protein of 43 kDa (TDP-43), a key factor in several neurodegenerative conditions, was discovered as a novel target of heteronemin by chemical proteomics. Combining bio-physical orthogonal approaches with biological analysis, heteronemin was found to influence the binding of TDP-43-cognate nucleic acids and to modulate the TDP-43 aggregation state and its cellular localization.

Cassiano C; Esposito R; Tosco A; Zampella A; D'Auria MV; Riccio R; Casapullo A; Monti MC

2013-08-01

 
 
 
 
61

Pathological mechanisms underlying TDP-43 driven neurodegeneration in FTLD-ALS spectrum disorders.  

UK PubMed Central (United Kingdom)

Aggregation of misfolded TAR DNA-binding protein 43 (TDP-43) is a striking hallmark of neurodegenerative processes that are observed in several neurological disorders, and in particular in most patients diagnosed with frontotemporal lobar degeneration (FTLD) or amyotrophic lateral sclerosis (ALS). A direct causal link with TDP-43 brain proteinopathy was provided by the identification of pathogenic mutations in TARDBP, the gene encoding TDP-43, in ALS families. However, TDP-43 proteinopathy has also been observed in carriers of mutations in several other genes associated with both ALS and FTLD demonstrating a key role for TDP-43 in neurodegeneration. To date, and despite substantial research into the biology of TDP-43, its functioning in normal brain and in neurodegeneration processes remains largely elusive. Nonetheless, breakthroughs using cellular and animal models have provided valuable insights into ALS and FTLD pathogenesis. Accumulating evidence has redirected the research focus towards a major role for impaired RNA metabolism and protein homeostasis. At the same time, the concept that toxic TDP-43 protein aggregates promote neurodegeneration is losing its credibility. This review aims at highlighting and discussing the current knowledge on TDP-43 driven pathomechanisms leading to neurodegeneration as observed in TDP-43 proteinopathies. Based on the complexity of the associated neurological diseases, a clear understanding of the essential pathological modifications will be crucial for further therapeutic interventions.

Janssens J; Van Broeckhoven C

2013-10-01

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TDP-43 skeins show properties of amyloid in a subset of ALS cases.  

UK PubMed Central (United Kingdom)

Aggregation of TDP-43 proteins to form intracellular inclusions is the primary pathology in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) with TDP-43 inclusions (FTLD-TDP). Histologically, in the cerebral cortex and limbic regions of affected ALS and FTLD-TDP patients, these pathologies occur as a variety of cytoplasmic, neuritic and intranuclear TDP-43 inclusions. In the spinal cord and lower brainstem of ALS patients, the lesions form cytoplasmic dashes or complex filamentous and spherical profiles in addition to skein-like inclusions (SLI). Ultrastructurally, the morphology of TDP-43 inclusions is heterogeneous but mainly composed of loose bundles of 10- to 20-nm-diameter straight filaments associated with electron-dense granular material. All of these TDP-43 inclusions are generally described as disordered amorphous aggregations unlike the amyloid fibrils that characterize protein accumulations in neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. We here report that Thioflavin-S positive SLI are present in a subset of ALS cases, while TDP-43 inclusions outside the spinal cord lack the chemical properties of amyloid. Further, we examine the differential enrichment of fibrillar profiles in SLI of ALS cases by TDP-43 immuno-electron microscopy (immuno-EM). The demonstration that pathological TDP-43 can be amyloidogenic in situ suggests the following conclusions: (1) the conformational changes associated with TDP-43 aggregation are more complex than previously thought; (2) Thioflavin-S positive SLI may be composed primarily of filamentous ultrastructures.

Robinson JL; Geser F; Stieber A; Umoh M; Kwong LK; Van Deerlin VM; Lee VM; Trojanowski JQ

2013-01-01

63

TDP-43 associates with stalled ribosomes and contributes to cell survival during cellular stress.  

UK PubMed Central (United Kingdom)

TAR DNA-binding protein 43 (TDP-43) has emerged as an important contributor to amyotrophic lateral sclerosis and frontotemporal lobar degeneration. To understand the physiological roles of TDP-43 in the complex translational regulation mechanisms, we exposed cultured cells to oxidative stress induced by sodium arsenite (ARS) for different periods of time, leading to non-lethal or sublethal injury. Polysome profile analysis revealed that ARS-induced stress caused the association of TDP-43 with stalled ribosomes via binding to mRNA, which was not found under the steady-state condition. When the cells were exposed to short-term/non-lethal stress, TDP-43 associating with ribosomes localized to stress granules (SGs); this association was transient because it was immediately dissolved by the removal of the stress. In contrast, when the cells were exposed to long-term/sublethal stress, TDP-43 was excluded from SGs and shifted to the heavy fractions independent of any binding to mRNA. In these severely stressed cells, biochemical alterations of TDP-43, such as increased insolubility and disulfide bond formation, were irreversible. TDP-43 was finally phosphorylated via the ARS-induced c-jun N-terminal kinase pathway. In TDP-43-silenced cells, stalled mRNA and poly (A)(+) RNA stability was disturbed and cytotoxicity increased under sublethal stress. Thus, TDP-43 associates with stalled ribosomes and contributes to cell survival during cellular stress.

Higashi S; Kabuta T; Nagai Y; Tsuchiya Y; Akiyama H; Wada K

2013-07-01

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

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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

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

UK PubMed Central (United Kingdom)

Abstract 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 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.

Hosokawa M; Arai T; Yamashita M; Tsuji H; Nonaka T; Masuda-Suzukake M; Tamaoka A; Hasegawa M; Akiyama H

2013-09-01

66

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

Science.gov (United States)

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 bridge this gap in our knowledge, we utilized novel cell culture and computer-assisted models to evaluate which region(s) of TDP-43 regulate its folding, self-interaction, biological activity and aggregation. We determined that the extreme N-terminus of TDP-43, specifically the first 10 residues, regulates folding of TDP-43 monomers necessary for proper homodimerization and TDP-43-regulated splicing. Despite such beneficial functions, we discovered an interesting dichotomy: full-length TDP-43 aggregation, which is believed to be a pathogenic process, also requires the extreme N-terminus of TDP-43. As such, we provide new insight into the structural basis for TDP-43 function and aggregation, and we suggest that stabilization of TDP-43 homodimers, the physiologically active form of TDP-43, may be a promising therapeutic strategy for ALS and FTLD-TDP.

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

67

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

UK PubMed Central (United Kingdom)

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 bridge this gap in our knowledge, we utilized novel cell culture and computer-assisted models to evaluate which region(s) of TDP-43 regulate its folding, self-interaction, biological activity and aggregation. We determined that the extreme N-terminus of TDP-43, specifically the first 10 residues, regulates folding of TDP-43 monomers necessary for proper homodimerization and TDP-43-regulated splicing. Despite such beneficial functions, we discovered an interesting dichotomy: full-length TDP-43 aggregation, which is believed to be a pathogenic process, also requires the extreme N-terminus of TDP-43. As such, we provide new insight into the structural basis for TDP-43 function and aggregation, and we suggest that stabilization of TDP-43 homodimers, the physiologically active form of TDP-43, may be a promising therapeutic strategy for ALS and FTLD-TDP.

Zhang YJ; Caulfield T; Xu YF; Gendron TF; Hubbard J; Stetler C; Sasaguri H; Whitelaw EC; Cai S; Lee WC; Petrucelli L

2013-08-01

68

Neuroprotection by Monocarbonyl Dimethoxycurcumin C: Ameliorating the Toxicity of Mutant TDP-43 via HO-1.  

UK PubMed Central (United Kingdom)

Mutation of TAR DNA-binding protein-43 (TDP-43) was detected in familiar and sporadic amyotrophic lateral sclerosis, and pathological TDP-43 was identified in the frontotemporal lobar degeneration. The neuroprotective functions of curcumin derivatives were assessed in motor neurons transfected with mutant TDP-43. We found that curcumin derivatives reduced the levels of TDP-43 fragments. Furthermore, we evaluated these compounds on the cellular model that the cells were transfected with TDP-25. We found that the expression level and aggregate formation of TDP-25 were significantly reduced by monocarbonyl dimethoxycurcumin C (Compound C). To study on the neuroprotective functions of curcumin derivatives, the neuroblastoma-spinal cord-34 cells transfected with mutant TDP-43 were assessed by the level of lactate dehydrogenase (LDH) and malondialdehyde bisdimethyl acetal (MDA) that were involved in the oxidative stress. We found that Compound C ameliorated the damage of mutant TDP-43 by reducing the level of MDA and LDH. Furthermore, heme oxygenase-1 (HO-1) was induced by Compound C significantly higher than other compounds. Znpp, which is known an inhibitor of HO-1, dramatically interfered with the function of Compound C. In addition, Compound C was tested in vivo, and HO-1 was significantly upregulated at the hippocampus. These findings suggest that Compound C, which degrades TDP-43 fragment and strengthens the antioxidant ability by HO-1, is a promising agent for TDP-43 proteinopathy.

Duan W; Guo Y; Xiao J; Chen X; Li Z; Han H; Li C

2013-08-01

69

TDP-43 Regulates the Microprocessor Complex Activity During In Vitro Neuronal Differentiation.  

UK PubMed Central (United Kingdom)

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.

Di Carlo V; Grossi E; Laneve P; Morlando M; Dini Modigliani S; Ballarino M; Bozzoni I; Caffarelli E

2013-10-01

70

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

UK PubMed Central (United Kingdom)

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.

Armakola M; Higgins MJ; Figley MD; Barmada SJ; Scarborough EA; Diaz Z; Fang X; Shorter J; Krogan NJ; Finkbeiner S; Farese RV Jr; Gitler AD

2012-12-01

71

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

2012-10-28

72

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.

Meyerowitz Jodi; Parker Sarah J; Vella Laura J; Ng Dominic CH; Price Katherine A; Liddell Jeffrey R; Caragounis Aphrodite; Li Qiao-Xin; Masters Colin L; Nonaka Takashi; Hasegawa Masato; Bogoyevitch Marie A; Kanninen Katja M; Crouch Peter J; White Anthony R

2011-01-01

73

Reduction of polyglutamine toxicity by TDP-43, FUS and progranulin in Huntington's disease models.  

UK PubMed Central (United Kingdom)

The DNA/RNA binding proteins TAR DNA-binding protein 43 (TDP-43) and fused-in-sarcoma (FUS) are genetically linked to amyotrophic lateral sclerosis and frontotemporal lobar dementia, while the inappropriate cytoplasmic accumulations of TDP-43 and FUS are observed in a growing number of late-onset pathologies including spinocerebellar ataxia 3, Alzheimer's and Huntington's diseases (HD). To investigate if TDP-43 and FUS contribute to neurodegenerative phenotypes, we turned to a genetically accessible Caenorhabditis elegans model of polyglutamine toxicity. In C. elegans, we observe that genetic loss-of-function mutations for nematode orthologs of TDP-43 or FUS reduced behavioral defects and neurodegeneration caused by huntingtin exon-1 with expanded polyglutamines. Furthermore, using striatal cells from huntingtin knock-in mice we observed that small interfering ribonucleic acid (siRNA) against TDP-43 or FUS reduced cell death caused by mutant huntingtin. Moreover, we found that TDP-43 and the survival factor progranulin (PGRN) genetically interact to regulate polyglutamine toxicity in C. elegans and mammalian cells. Altogether our data point towards a conserved function for TDP-43 and FUS in promoting polyglutamine toxicity and that delivery of PGRN may have therapeutic benefits.

Tauffenberger A; Chitramuthu BP; Bateman A; Bennett HP; Parker JA

2013-02-01

74

The JNK/c-Jun signaling axis contributes to the TDP-43-induced cell death.  

UK PubMed Central (United Kingdom)

Dysregulation of transactive response DNA-binding protein-43 (TDP-43) is closely linked to the pathogenesis of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U). The contribution of the upregulation of TDP-43 expression to the pathogenesis has been strongly suggested by the observation that the level of TDP-43 expression is increased in both ALS and FTLD-U patients. We previously found that the low-grade (twice to five times more than the endogenous level) overexpression of TDP-43 induces neuronal cell death through the upregulation of Bim and CHOP expression and the downregulation of Bcl-xL expression. In this study, we further show that the low-grade overexpression of TDP-43 increases the level of phosphorylated c-Jun N-terminal kinase (JNK) and the co-incubation with a JNK inhibitor, the expression of a dominant-negative JNK, or the expression of a dominant-negative c-Jun inhibited the TDP-43-induced death in NSC34 motor neuronal cells. These data together suggest that the JNK/c-Jun signaling axis contributes to the TDP-43-induced cell death.

Suzuki H; Matsuoka M

2013-01-01

75

Pharmacological reduction of ER stress protects against TDP-43 neuronal toxicity in vivo.  

UK PubMed Central (United Kingdom)

C. elegans and D. rerio expressing mutant TAR DNA Binding Protein 43 (TDP-43) are powerful in vivo animal models for the genetics and pharmacology of amyotrophic lateral sclerosis (ALS). Using these small-animal models of ALS, we previously identified methylene blue (MB) as a potent suppressor of TDP-43 toxicity. Consequently here we investigated how MB might exert its neuroprotective properties and found that it acts through reduction of the endoplasmic reticulum (ER) stress response. We tested other compounds known to be active in the ER unfolded protein response in worms and zebrafish expressing mutant human TDP-43 (mTDP-43). We identified three compounds: salubrinal, guanabenz and a new structurally related compound phenazine, which also reduced paralysis, neurodegeneration and oxidative stress in our mTDP-43 models. Using C. elegans genetics, we showed that all four compounds act as potent suppressors of mTDP-43 toxicity through reduction of the ER stress response. Interestingly, these compounds operate through different branches of the ER unfolded protein pathway to achieve a common neuroprotective action. Our results indicate that protein-folding homeostasis in the ER is an important target for therapeutic development in ALS and other TDP-43-related neurodegenerative diseases.

Vaccaro A; Patten SA; Aggad D; Julien C; Maios C; Kabashi E; Drapeau P; Parker JA

2013-07-01

76

TDP-43, an ALS Linked Protein, Regulates Fat Deposition and Glucose Homeostasis  

Science.gov (United States)

The identification of proteins which determine fat and lean body mass composition is critical to better understanding and treating human obesity. TDP-43 is a well-conserved RNA-binding protein known to regulate alternative splicing and recently implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS). While TDP-43 knockout mice show early embryonic lethality, post-natal conditional knockout mice show weight loss, fat depletion, and rapid death, suggesting an important role for TDP-43 in regulating energy metabolism. Here we report, that over-expression of TDP-43 in transgenic mice can result in a phenotype characterized by increased fat deposition and adipocyte hypertrophy. In addition, TDP-43 over-expression in skeletal muscle results in increased steady state levels of Tbc1d1, a RAB-GTPase activating protein involved in Glucose 4 transporter (Glut4) translocation. Skeletal muscle fibers isolated from TDP-43 transgenic mice show altered Glut4 translocation in response to insulin and impaired insulin mediated glucose uptake. These results indicate that levels of TDP-43 regulate body fat composition and glucose homeostasis in vivo.

Stallings, Nancy R.; Puttaparthi, Krishna; Dowling, Katherine J.; Luther, Christina M.; Burns, Dennis K.; Davis, Kathryn; Elliott, Jeffrey L.

2013-01-01

77

TDP-43, an ALS linked protein, regulates fat deposition and glucose homeostasis.  

UK PubMed Central (United Kingdom)

The identification of proteins which determine fat and lean body mass composition is critical to better understanding and treating human obesity. TDP-43 is a well-conserved RNA-binding protein known to regulate alternative splicing and recently implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS). While TDP-43 knockout mice show early embryonic lethality, post-natal conditional knockout mice show weight loss, fat depletion, and rapid death, suggesting an important role for TDP-43 in regulating energy metabolism. Here we report, that over-expression of TDP-43 in transgenic mice can result in a phenotype characterized by increased fat deposition and adipocyte hypertrophy. In addition, TDP-43 over-expression in skeletal muscle results in increased steady state levels of Tbc1d1, a RAB-GTPase activating protein involved in Glucose 4 transporter (Glut4) translocation. Skeletal muscle fibers isolated from TDP-43 transgenic mice show altered Glut4 translocation in response to insulin and impaired insulin mediated glucose uptake. These results indicate that levels of TDP-43 regulate body fat composition and glucose homeostasis in vivo.

Stallings NR; Puttaparthi K; Dowling KJ; Luther CM; Burns DK; Davis K; Elliott JL

2013-01-01

78

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.

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

2011-01-01

79

[TDP-43 proteinopathies - from frontotemporal lobar degeneration to inclusion body myositis].  

UK PubMed Central (United Kingdom)

TDP-43, a newly described neurodegenerative protein, is of great interest to both neurologists and geneticists. At the beginning, its dysfunction was recognized in sporadic amyotrophic lateral sclerosis, frontotemporal lobar degeneration with ubiquitinated inclusions and in mixed forms. However, it was also proved that TDP-43 inclusions are in addition present in many other diseases, for example in inclusion body myositis. Furthermore, many genes and different loci may be involved in pathological TDP-43 accumulation in cells and tissues. Mutations in the TARDPB gene, progranulin gene (PGRNVCP) as well as a gene on chromosome 9p were found. The present paper is a summary on possible involvement of TDP-43 in various neurodegenerative disorders.

Kierdaszuk B; Berdy?ski M; Zekanowski C; Kami?ska A

2012-07-01

80

TDP-43 proteinopathy and motor neuron disease in chronic traumatic encephalopathy.  

Science.gov (United States)

Epidemiological evidence suggests that the incidence of amyotrophic lateral sclerosis is increased in association with head injury. Repetitive head injury is also associated with the development of chronic traumatic encephalopathy (CTE), a tauopathy characterized by neurofibrillary tangles throughout the brain in the relative absence of ?-amyloid deposits. We examined 12 cases of CTE and, in 10, found a widespread TAR DNA-binding protein of approximately 43kd (TDP-43) proteinopathy affecting the frontal and temporal cortices, medial temporal lobe, basal ganglia, diencephalon, and brainstem. Three athletes with CTE also developed a progressive motor neuron disease with profound weakness, atrophy, spasticity, and fasciculations several years before death. In these 3 cases, there were abundant TDP-43-positive inclusions and neurites in the spinal cord in addition to tau neurofibrillary changes, motor neuron loss, and corticospinal tract degeneration. The TDP-43 proteinopathy associated with CTE is similar to that found in frontotemporal lobar degeneration with TDP-43 inclusions, in that widespread regions of the brain are affected. Akin to frontotemporal lobar degeneration with TDP-43 inclusions, in some individuals with CTE, the TDP-43 proteinopathy extends to involve the spinal cord and is associated with motor neuron disease. This is the first pathological evidence that repetitive head trauma experienced in collision sports might be associated with the development of a motor neuron disease. PMID:20720505

McKee, Ann C; Gavett, Brandon E; Stern, Robert A; Nowinski, Christopher J; Cantu, Robert C; Kowall, Neil W; Perl, Daniel P; Hedley-Whyte, E Tessa; Price, Bruce; Sullivan, Chris; Morin, Peter; Lee, Hyo-Soon; Kubilus, Caroline A; Daneshvar, Daniel H; Wulff, Megan; Budson, Andrew E

2010-09-01

 
 
 
 
81

TDP-43 proteinopathy and motor neuron disease in chronic traumatic encephalopathy.  

UK PubMed Central (United Kingdom)

Epidemiological evidence suggests that the incidence of amyotrophic lateral sclerosis is increased in association with head injury. Repetitive head injury is also associated with the development of chronic traumatic encephalopathy (CTE), a tauopathy characterized by neurofibrillary tangles throughout the brain in the relative absence of ?-amyloid deposits. We examined 12 cases of CTE and, in 10, found a widespread TAR DNA-binding protein of approximately 43kd (TDP-43) proteinopathy affecting the frontal and temporal cortices, medial temporal lobe, basal ganglia, diencephalon, and brainstem. Three athletes with CTE also developed a progressive motor neuron disease with profound weakness, atrophy, spasticity, and fasciculations several years before death. In these 3 cases, there were abundant TDP-43-positive inclusions and neurites in the spinal cord in addition to tau neurofibrillary changes, motor neuron loss, and corticospinal tract degeneration. The TDP-43 proteinopathy associated with CTE is similar to that found in frontotemporal lobar degeneration with TDP-43 inclusions, in that widespread regions of the brain are affected. Akin to frontotemporal lobar degeneration with TDP-43 inclusions, in some individuals with CTE, the TDP-43 proteinopathy extends to involve the spinal cord and is associated with motor neuron disease. This is the first pathological evidence that repetitive head trauma experienced in collision sports might be associated with the development of a motor neuron disease.

McKee AC; Gavett BE; Stern RA; Nowinski CJ; Cantu RC; Kowall NW; Perl DP; Hedley-Whyte ET; Price B; Sullivan C; Morin P; Lee HS; Kubilus CA; Daneshvar DH; Wulff M; Budson AE

2010-09-01

82

TDP-43 pathology in polyglutamine diseases: With reference to amyotrphic lateral sclerosis.  

UK PubMed Central (United Kingdom)

A nuclear protein, transactivation response (TAR) DNA binding protein 43?kDa (TDP-43), is the major component of neuronal cytoplasmic inclusions (NCIs) in frontotemporal lobar degeneration with ubiquitin inclusions (FTLD-U) and sporadic amyotrophic lateral sclerosis (SALS). While initially thought to be relatively specific to FTLD-U and ALS, TDP-43 pathology has now been detected in a number of other neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease. In such tauopathies and ?-synucleinopathies, occurrence of TDP-43-positive neuronal cytoplasmic inclusions may be associated with other distinct molecular pathologic processes primarily involving their own pathological proteins, tau and ?-synuclein, respectively (secondary TDP-43 proteinopathies). On the other hand, in several polyglutamine (polyQ) diseases, TDP-43 appears to play an important pathomechanistic role. Interestingly, intermediate-length polyQ expansions (27-33 Qs) in ataxin 2, the causative gene of spinocerebellar ataxia type 2, have recently been reported to be a genetic risk factor for SALS. Here, with a review of the literature, we discuss the relationship between ALS and polyQ diseases from the viewpoint of TDP-43 neuropathology.

Toyoshima Y; Takahashi H

2013-07-01

83

Accumulation of phosphorylated TDP-43 in the CNS of a patient with Cockayne syndrome.  

UK PubMed Central (United Kingdom)

Here, we report a case of Cockayne syndrome (CS) in a Japanese man who displayed a unique pathology of phosphorylated trans-activation response (TAR) DNA-binding protein 43 (pTDP-43) with abundant Rosenthal fibers. Many round pTDP-43-positive structures were detected throughout the CNS; however, most of them were located in two regions that also exhibited neuronal depletion: the cerebellar cortex and the inferior olivary nucleus. To a lesser extent, these aggregates were also present in the cerebellar white matter, around the subependymal regions in the brain stem, and in the spinal cord. Intraneuronal pTDP-43 inclusions were only observed in a small number of neurons in the inferior olivary nucleus. Double-label immunofluorescence revealed that many of the aggregates were localized to astrocytes. The observed distribution and the morphology of the pTDP-43-positive structures were unique and have not yet been reported. Therefore, a pTDP-43-related pathology may be implicated in CS as well as in other neurodegenerative diseases such as frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Whether the pathology of these diseases reflects a primary neurodegenerative process or a secondary reaction is not known.

Sakurai A; Makioka K; Fukuda T; Takatama M; Okamoto K

2013-04-01

84

Wild type TDP-43 induces neuro-inflammation and alters APP metabolism in lentiviral gene transfer models.  

UK PubMed Central (United Kingdom)

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 determine whether there is a relationship between TDP-43 and APP metabolism. We generated gene transfer models using lentiviral delivery of human TDP-43 and A?(1-42) into the rat primary motor cortex and examined their role 2 weeks post-injection. Expression of TDP-43 and/or A?(1-42) increase pro-inflammatory markers, including Interleukin (IL)-6, tumor necrosis factor (TNF-?), glial neurofibrillary proteins (GFAP) and ionized calcium binding adaptor molecule 1 (IBA-1). Lentiviral A?(1-42) up-regulates endogenous TDP-43 and promotes its phosphorylation, aggregation and cleavage into 35 kDa fragments. Inversely, lentiviral TDP-43 expression increases the levels and activity of ?-secretase (BACE), accelerating production of APP C-terminal fragments (C99) and A?(1-40). Here we show that TDP-43 up-regulates APP metabolism and suggest a mechanistic link between TDP-43 and BACE.

Herman AM; Khandelwal PJ; Rebeck GW; Moussa CE

2012-05-01

85

Motor neurons and glia exhibit specific individualized responses to TDP-43 expression in a Drosophila model of amyotrophic lateral sclerosis.  

UK PubMed Central (United Kingdom)

Amyotrophic lateral sclerosis (ALS) is a fatal disease characterized by complex neuronal and glial phenotypes. Recently, RNA-based mechanisms have been linked to ALS via RNA-binding proteins such as TDP-43, which has been studied in vivo using models ranging from yeast to rodents. We have developed a Drosophila model of ALS based on TDP-43 that recapitulates several aspects of pathology, including motor neuron loss, locomotor dysfunction and reduced survival. Here we report the phenotypic consequences of expressing wild-type and four different ALS-linked TDP-43 mutations in neurons and glia. We show that TDP-43-driven neurodegeneration phenotypes are dose- and age-dependent. In motor neurons, TDP-43 appears restricted to nuclei, which are significantly misshapen due to mutant but not wild-type protein expression. In glia and in the developing neuroepithelium, TDP-43 associates with cytoplasmic puncta. TDP-43-containing RNA granules are motile in cultured motor neurons, although wild-type and mutant variants exhibit different kinetic properties. At the neuromuscular junction, the expression of TDP-43 in motor neurons versus glia leads to seemingly opposite synaptic phenotypes that, surprisingly, translate into comparable locomotor defects. Finally, we explore sleep as a behavioral readout of TDP-43 expression and find evidence of sleep fragmentation consistent with hyperexcitability, a suggested mechanism in ALS. These findings support the notion that although motor neurons and glia are both involved in ALS pathology, at the cellular level they can exhibit different responses to TDP-43. In addition, our data suggest that individual TDP-43 alleles utilize distinct molecular mechanisms, which will be important for developing therapeutic strategies.

Estes PS; Daniel SG; McCallum AP; Boehringer AV; Sukhina AS; Zwick RA; Zarnescu DC

2013-05-01

86

Parkin ubiquitinates Tar-DNA binding protein-43 (TDP-43) and promotes its cytosolic accumulation via interaction with histone deacetylase 6 (HDAC6).  

UK PubMed Central (United Kingdom)

The importance of E3 ubiquitin ligases, involved in the degradation of misfolded proteins or promotion of protein-protein interaction, is increasingly recognized in neurodegeneration. TDP-43 is a predominantly nuclear protein, which regulates the transcription of thousands of genes and binds to mRNA of the E3 ubiquitin ligase Parkin to regulate its expression. Wild type and mutated TDP-43 are detected in ubiquitinated forms within the cytosol in several neurodegenerative diseases. We elucidated the mechanisms of TDP-43 interaction with Parkin using transgenic A315T mutant TDP-43 (TDP43-Tg) mice, lentiviral wild type TDP-43, and Parkin gene transfer rat models. TDP-43 expression increased Parkin mRNA and protein levels. Lentiviral TDP-43 increased the levels of nuclear and cytosolic protein, whereas Parkin co-expression mediated Lys-48 and Lys-63-linked ubiquitin to TDP-43 and led to cytosolic co-localization of Parkin with ubiquitinated TDP-43. Parkin and TDP-43 formed a multiprotein complex with HDAC6, perhaps to mediate TDP-43 translocation. In conclusion, Parkin ubiquitinates TDP-43 and facilitates its cytosolic accumulation through a multiprotein complex with HDAC6.

Hebron ML; Lonskaya I; Sharpe K; Weerasinghe PP; Algarzae NK; Shekoyan AR; Moussa CE

2013-02-01

87

A High-Content Screen Identifies Novel Compounds That Inhibit Stress-Induced TDP-43 Cellular Aggregation and Associated Cytotoxicity.  

UK PubMed Central (United Kingdom)

TDP-43 is an RNA binding protein found to accumulate in the cytoplasm of brain and spinal cord from patients affected with amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Nuclear TDP-43 protein regulates transcription through several mechanisms, and under stressed conditions, it forms cytoplasmic aggregates that co-localize with stress granule (SG) proteins in cell culture. These granules are also found in the brain and spinal cord of patients affected with ALS and FTLD. The mechanism through which TDP-43 might contribute to neurodegenerative diseases is poorly understood. To investigate the pathophysiology of TDP-43 aggregation and to isolate potential therapeutic targets, we screened a chemical library of 75,000 compounds using high-content analysis with PC12 cells that inducibly express human TDP-43 tagged with green fluorescent protein (GFP). The screen identified 16 compounds that dose-dependently decreased the TDP-43 inclusions without significant cellular toxicity or changes in total TDP-43 expression levels. To validate the effect, we tested compounds by Western blot analysis and in a Caenorhabditis elegans model that replicates some of the relevant disease phenotypes. The hits from this assay will be useful for elucidating regulation of TDP-43, stress granule response, and possible ALS therapeutics.

Boyd JD; Lee-Armandt JP; Feiler MS; Zauur N; Liu M; Kraemer B; Concannon JB; Ebata A; Wolozin B; Glicksman MA

2013-09-01

88

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

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...

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

89

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

Science.gov (United States)

TDP-43 is a critical RNA-binding factor associated with pre-mRNA splicing in mammals. Its expression is tightly autoregulated, with loss of this regulation implicated in human neuropathology. We demonstrate that TDP-43 overexpression in humans and mice activates a 3? untranslated region (UTR) intron, resulting in excision of the proximal polyA site (PAS) pA1. This activates a cryptic PAS that prevents TDP-43 expression through a nuclear retention mechanism. Superimposed on this process, overexpression of TDP-43 blocks recognition of pA1 by competing with CstF-64 for PAS binding. Overall, we uncover complex interplay between transcription, splicing, and 3? end processing to effect autoregulation of TDP-43.

Avendano-Vazquez, S. Erendira; Dhir, Ashish; Bembich, Sara; Buratti, Emanuele; Proudfoot, Nicholas; Baralle, Francisco E.

2012-01-01

90

TDP-43 is a culprit in human neurodegeneration, and not just an innocent bystander  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In 2006 the protein TDP-43 was identified as the major ubiquitinated component deposited in the inclusion bodies found in two human neurodegenerative diseases, amyotrophic lateral sclerosis and frontotemporal lobar degeneration. The pathogenesis of both disorders is unclear, although they are relate...

Banks, Gareth T.; Kuta, Anna; Isaacs, Adrian M.; Fisher, Elizabeth M. C.

91

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

Directory of Open Access Journals (Sweden)

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 Anaďs; Stabile Stéphanie; Vande Velde Christine

2012-01-01

92

Co-localization of Bunina bodies and TDP-43 inclusions in lower motor neurons in amyotrophic lateral sclerosis.  

UK PubMed Central (United Kingdom)

Amyotrophic lateral sclerosis (ALS) is characterized by motor neuron involvement with Bunina bodies (BBs) and transactivation response DNA protein 43 (TDP-43) inclusions. We examined the spinal cord (n?=?20), hypoglossal nucleus (n?=?6) and facial nucleus (n?=?5) from ALS patients to elucidate the relationship between BBs and TDP-43 inclusions. BBs were found in the anterior horn in 16 of 20 cases, in the hypoglossal nucleus in all six cases and in the facial nucleus in four out of five cases. TDP-43 inclusions were found in each region of all the cases. Co-localization of BBs and TDP-43 inclusions was found in 15.2% of total neurons in the anterior horn, 29.2% in the hypoglossal nucleus and 17.3% in the facial nucleus. The frequency of TDP-43 inclusions was significantly higher in neurons with BBs than in those without in each region. Ultrastructurally, TDP-43-positive filamentous structures were intermingled with BBs. These findings suggest that there is a close relationship in the occurrence between BBs and TDP-43 inclusions.

Mori F; Kakita A; Takahashi H; Wakabayashi K

2013-05-01

93

The FTD/ALS-associated RNA-binding protein TDP-43 regulates the robustness of neuronal specification through microRNA-9a in Drosophila.  

UK PubMed Central (United Kingdom)

TDP-43 is an evolutionarily conserved RNA-binding protein currently under intense investigation for its involvement in the molecular pathogenesis of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). TDP-43 is normally localized in the nucleus, but translocated to the cytoplasm in diseased neurons. The endogenous functions of TDP-43 in the nervous system remain poorly understood. Here, we show that the loss of Drosophila TDP-43 (dTDP-43) results in an increased production of sensory bristles and sensory organ precursor (SOP) cells on the notum of some but not all flies. The location of ectopic SOPs varies among mutant flies. The penetrance of this novel phenotype is dependent on the gender and sensitive to environmental influences. A similar SOP phenotype was also observed on the wing and in the embryos. Overexpression of dTDP-43 causes both loss and ectopic production of SOPs. Ectopic expression of ALS-associated mutant human TDP-43 (hTDP-43(M337V) and hTDP-43(Q331K)) produces a less severe SOP phenotype than hTDP-43(WT), indicating a partial loss of function of mutant hTDP-43. In dTDP-43 mutants, miR-9a expression is significantly reduced. Genetic interaction studies further support the notion that dTDP-43 acts through miR-9a to control the precision of SOP specification. These findings reveal a novel role for endogenous TDP-43 in neuronal specification and suggest that the FTD/ALS-associated RNA-binding protein TDP-43 functions to ensure the robustness of genetic control programs.

Li Z; Lu Y; Xu XL; Gao FB

2013-01-01

94

Accumulation of C-terminal fragments of transactive response DNA-binding protein 43 leads to synaptic loss and cognitive deficits in human TDP-43 transgenic mice.  

UK PubMed Central (United Kingdom)

Accumulation of the transactive response DNA-binding protein 43 (TDP-43) is a major hallmark of several neurodegenerative disorders, collectively known as TDP-43 proteinopathies. The most common TDP-43 proteinopathies, frontotemporal lobar degeneration with TDP-43-positive inclusions, and amyotrophic lateral sclerosis, share overlapping neuropathological and clinical phenotypes. The development and detailed analysis of animal models of TDP-43 proteinopathies are critical for understanding the pathogenesis of these disorders. Transgenic mice overexpressing mutant human TDP-43 (herein referred to as hTDP-43) are characterized by neurodegeneration and reduced life span. However, little is known about the behavioral phenotype of these mice. Here we report the novel finding that hTDP-43 mice develop deficits in cognition, motor performance, and coordination. We show that these behavioral deficits are associated with the accumulation of nuclear and cytosolic TDP-43 C-terminal fragments, a decrease in endogenous TDP-43 levels, and synaptic loss. Our findings provide critical insights into disease pathology, and will help guide future preclinical studies aimed at testing the effects of potential therapeutic agents on the onset and progression of TDP-43 proteinopathies.

Medina DX; Orr ME; Oddo S

2014-01-01

95

Structural transformation of the amyloidogenic core region of TDP-43 protein initiates its aggregation and cytoplasmic inclusion.  

UK PubMed Central (United Kingdom)

TDP-43 (TAR DNA-binding protein of 43 kDa) is a major deposited protein in amyotrophic lateral sclerosis and frontotemporal dementia with ubiquitin. A great number of genetic mutations identified in the flexible C-terminal region are associated with disease pathologies. We investigated the molecular determinants of TDP-43 aggregation and its underlying mechanisms. We identified a hydrophobic patch (residues 318-343) as the amyloidogenic core essential for TDP-43 aggregation. Biophysical studies demonstrated that the homologous peptide formed a helix-turn-helix structure in solution, whereas it underwent structural transformation from an ?-helix to a ?-sheet during aggregation. Mutation or deletion of this core region significantly reduced the aggregation and cytoplasmic inclusions of full-length TDP-43 (or TDP-35 fragment) in cells. Thus, structural transformation of the amyloidogenic core initiates the aggregation and cytoplasmic inclusion formation of TDP-43. This particular core region provides a potential therapeutic target to design small-molecule compounds for mitigating TDP-43 proteinopathies.

Jiang LL; Che MX; Zhao J; Zhou CJ; Xie MY; Li HY; He JH; Hu HY

2013-07-01

96

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

Directory of Open Access Journals (Sweden)

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.

Caccamo Antonella; Magrí Andrea; Oddo Salvatore

2010-01-01

97

Similar dose-dependence of motor neuron cell death caused by wild type human TDP-43 and mutants with ALS-associated amino acid substitutions  

Science.gov (United States)

Background TDP-43, a multi-functional DNA/ RNA-binding protein encoded by the TARDBP gene, has emerged as a major patho-signature factor of the ubiquitinated intracellular inclusions (UBIs) in the diseased cells of a range of neurodegenerative diseases. Mutations in at least 9 different genes including TARDBP have been identified in ALS with TDP-43 (+)-UBIs. Thus far, the pathogenic role(s) of the more than 30 ALS-associated mutations in the TARDBP gene has not been well defined. Results By transient DNA transfection studies, we show that exogenously expressed human TDP-43 (hTDP-43), either wild type (WT) or 2 different ALS mutant (MT) forms, could cause significantly higher apoptotic death rate of a mouse spinal motor neuron-like cell line (NSC34) than other types of cells, e.g. mouse neuronal Neuro2a and human fibroblast HEK293T cells. Furthermore, at the same plasmid DNA dose(s) used for transfection, the percentages of NSC34 cell death caused by the 2 exogenously expressed hTDP-43 mutants are all higher than that caused by the WT hTDP-43. Significantly, the above observations are correlated with higher steady-state levels of the mutant hTDP-43 proteins as well as their stabilities than the WT. Conclusions Based on these data and previous transgenic TDP-43 studies in animals or cell cultures, we suggest that one major common consequence of the different ALS-associated TDP-43 mutations is the stabilization of the hTDP-43 polypeptide. The resulting elevation of the steady state level of hTDP-43 in combination with the relatively low tolerance of the spinal motor neurons to the increased amount of hTDP-43 lead to the neurodegeneration and pathogenesis of ALS, and of diseases with TDP-43 proteinopathies in general.

2013-01-01

98

Similar dose-dependence of motor neuron cell death caused by wild type human TDP-43 and mutants with ALS-associated amino acid substitutions.  

UK PubMed Central (United Kingdom)

BACKGROUND: TDP-43, a multi-functional DNA/ RNA-binding protein encoded by the TARDBP gene, has emerged as a major patho-signature factor of the ubiquitinated intracellular inclusions (UBIs) in the diseased cells of a range of neurodegenerative diseases. Mutations in at least 9 different genes including TARDBP have been identified in ALS with TDP-43 (+)-UBIs. Thus far, the pathogenic role(s) of the more than 30 ALS-associated mutations in the TARDBP gene has not been well defined. RESULTS: By transient DNA transfection studies, we show that exogenously expressed human TDP-43 (hTDP-43), either wild type (WT) or 2 different ALS mutant (MT) forms, could cause significantly higher apoptotic death rate of a mouse spinal motor neuron-like cell line (NSC34) than other types of cells, e.g. mouse neuronal Neuro2a and human fibroblast HEK293T cells. Furthermore, at the same plasmid DNA dose(s) used for transfection, the percentages of NSC34 cell death caused by the 2 exogenously expressed hTDP-43 mutants are all higher than that caused by the WT hTDP-43. Significantly, the above observations are correlated with higher steady-state levels of the mutant hTDP-43 proteins as well as their stabilities than the WT. CONCLUSIONS: Based on these data and previous transgenic TDP-43 studies in animals or cell cultures, we suggest that one major common consequence of the different ALS-associated TDP-43 mutations is the stabilization of the hTDP-43 polypeptide. The resulting elevation of the steady state level of hTDP-43 in combination with the relatively low tolerance of the spinal motor neurons to the increased amount of hTDP-43 lead to the neurodegeneration and pathogenesis of ALS, and of diseases with TDP-43 proteinopathies in general.

Wu LS; Cheng WC; Shen CK

2013-01-01

99

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

UK PubMed Central (United Kingdom)

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, subcellular mislocalization of TDP-43, and decreased cell survival. We then performed coculture experiments to evaluate the effects of M337V astrocytes on the survival of wild-type and M337V TDP-43 motor neurons, showing that mutant TDP-43 astrocytes do not adversely affect survival of cocultured neurons. These observations reveal a significant and previously unrecognized glial cell-autonomous pathological phenotype associated with a pathogenic mutation in TDP-43 and show that TDP-43 proteinopathies do not display an astrocyte non-cell-autonomous component in cell culture, as previously described for SOD1 ALS. This study highlights the utility of induced pluripotent stem cell-based in vitro disease models to investigate mechanisms of disease in ALS and other TDP-43 proteinopathies.

Serio A; Bilican B; Barmada SJ; Ando DM; Zhao C; Siller R; Burr K; Haghi G; Story D; Nishimura AL; Carrasco MA; Phatnani HP; Shum C; Wilmut I; Maniatis T; Shaw CE; Finkbeiner S; Chandran S

2013-03-01

100

Coaggregation of RNA-binding proteins in a model of TDP-43 proteinopathy with selective RGG motif methylation and a role for RRM1 ubiquitination.  

UK PubMed Central (United Kingdom)

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 following overexpression, using stable isotope-labeled (SILAC) internal standards and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). We also searched for differential post-translational modification (PTM) sites of ubiquitination. Four sites of ubiquitin conjugation to TDP-43 or TDP-S6 were confirmed by dialkylated GST-TDP-43 external reference peptides, occurring on or near RNA binding motif (RRM) 1. RRM-containing proteins co-enriched in cytoplasmic granular structures in HEK-293 cells and primary motor neurons with insoluble TDP-S6, including cytoplasmic stress granule associated proteins G3BP, PABPC1, and eIF4A1. Proteomic evidence for TDP-43 co-aggregation with paraspeckle markers RBM14, PSF and NonO was also validated by western blot and by immunocytochemistry in HEK-293 cells. An increase in peptides from methylated arginine-glycine-glycine (RGG) RNA-binding motifs of FUS/TLS and hnRNPs was found in the detergent-insoluble fraction of TDP-overexpressing cells. Finally, TDP-43 and TDP-S6 detergent-insoluble species were reduced by mutagenesis of the identified ubiquitination sites, even following oxidative or proteolytic stress. Together, these findings define some of the aggregation partners of TDP-43, and suggest that TDP-43 ubiquitination influences TDP-43 oligomerization.

Dammer EB; Fallini C; Gozal YM; Duong DM; Rossoll W; Xu P; Lah JJ; Levey AI; Peng J; Bassell GJ; Seyfried NT

2012-01-01

 
 
 
 
101

Selective forelimb impairment in rats expressing a pathological TDP-43 25?kDa C-terminal fragment to mimic amyotrophic lateral sclerosis.  

UK PubMed Central (United Kingdom)

Pathological inclusions containing transactive response DNA-binding protein 43?kDa (TDP-43) are common in several neurodegenerative diseases including amyotrophic lateral sclerosis (ALS). TDP-43 normally localizes predominantly to the nucleus, but during disease progression, it mislocalizes to the cytoplasm. We expressed TDP-43 in rats by an adeno-associated virus (AAV9) gene transfer method that transduces neurons throughout the central nervous system (CNS). To mimic the aberrant cytoplasmic TDP-43 found in disease, we expressed a form of TDP-43 with mutations in the nuclear localization signal sequence (TDP-NLS). The TDP-NLS was detected in both the cytoplasm and the nucleus of transduced neurons. Unlike wild-type TDP-43, expression of TDP-NLS did not induce mortality. However, the TDP-NLS induced disease-relevant motor impairments over 24 weeks. We compared the TDP-NLS to a 25?kDa C-terminal proaggregatory fragment of TDP-43 (TDP-25). The clinical phenotype of forelimb impairment was pronounced with the TDP-25 form, supporting a role of this C-terminal fragment in pathogenesis. The results advance previous rodent models by inducing cytoplasmic expression of TDP-43 in the spinal cord, and the non-lethal phenotype enabled long-term study. Approaching a more relevant disease state in an animal model that more closely mimics underlying mechanisms in human disease could unlock our ability to develop therapeutics.

Dayton RD; Gitcho MA; Orchard EA; Wilson JD; Wang DB; Cain CD; Johnson JA; Zhang YJ; Petrucelli L; Mathis JM; Klein RL

2013-07-01

102

Transcriptome-wide analysis of TDP-43 binding small RNAs identifies miR-NID1, a novel miRNA that represses NRXN1 expression.  

UK PubMed Central (United Kingdom)

The Tar DNA-binding protein 43 (TDP-43) regulates RNA processing and miRNA biogenesis and is known to be involved in neurodegeneration. Messenger RNA (mRNA) targets of TDP-43 have recently been systematically identified, but small RNAs (sRNAs) bound by TDP-43 have not been studied in details. Here, we reexamine cross-linking, immunoprecipitation and sequencing (CLIP-seq) data, and identify pre-miRNAs, miRNAs and piRNAs bound by TDP-43 in human and mouse brains. Subsequent analysis of TDP-43 binding miRNAs suggests that target genes are enriched in functions involving synaptic activities. We further identify a novel miRNA (miR-NID1) processed from the intron 5 of human neurexin 1, NRXN1, and show that miR-NID1 represses NRXN1 expression by binding to TDP-43. Our results are in accordance with previously published data indicating TDP-43 through binding of specific miRNAs to play roles in neurodevelopmental activities and neurological disorders and further our understanding of TDP-43 function.

Fan Z; Chen X; Chen R

2013-07-01

103

Loss of ALS-associated TDP-43 in zebrafish causes muscle degeneration, vascular dysfunction, and reduced motor neuron axon outgrowth.  

UK PubMed Central (United Kingdom)

Mutations in the Tar DNA binding protein of 43 kDa (TDP-43; TARDBP) are associated with amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with TDP-43(+) inclusions (FTLD-TDP). To determine the physiological function of TDP-43, we knocked out zebrafish Tardbp and its paralogue Tardbp (TAR DNA binding protein-like), which lacks the glycine-rich domain where ALS- and FTLD-TDP-associated mutations cluster. tardbp mutants show no phenotype, a result of compensation by a unique splice variant of tardbpl that additionally contains a C-terminal elongation highly homologous to the glycine-rich domain of tardbp. Double-homozygous mutants of tardbp and tardbpl show muscle degeneration, strongly reduced blood circulation, mispatterning of vessels, impaired spinal motor neuron axon outgrowth, and early death. In double mutants the muscle-specific actin binding protein Filamin Ca is up-regulated. Strikingly, Filamin C is similarly increased in the frontal cortex of FTLD-TDP patients, suggesting aberrant expression in smooth muscle cells and TDP-43 loss-of-function as one underlying disease mechanism.

Schmid B; Hruscha A; Hogl S; Banzhaf-Strathmann J; Strecker K; van der Zee J; Teucke M; Eimer S; Hegermann J; Kittelmann M; Kremmer E; Cruts M; Solchenberger B; Hasenkamp L; van Bebber F; Van Broeckhoven C; Edbauer D; Lichtenthaler SF; Haass C

2013-03-01

104

The ALS-associated proteins FUS and TDP-43 function together to affect Drosophila locomotion and life span.  

UK PubMed Central (United Kingdom)

The fatal adult motor neuron disease amyotrophic lateral sclerosis (ALS) shares some clinical and pathological overlap with frontotemporal dementia (FTD), an early-onset neurodegenerative disorder. The RNA/DNA-binding proteins fused in sarcoma (FUS; also known as TLS) and TAR DNA binding protein-43 (TDP-43) have recently been shown to be genetically and pathologically associated with familial forms of ALS and FTD. It is currently unknown whether perturbation of these proteins results in disease through mechanisms that are independent of normal protein function or via the pathophysiological disruption of molecular processes in which they are both critical. Here, we report that Drosophila mutants in which the homolog of FUS is disrupted exhibit decreased adult viability, diminished locomotor speed, and reduced life span compared with controls. These phenotypes were fully rescued by wild-type human FUS, but not ALS-associated mutant FUS proteins. A mutant of the Drosophila homolog of TDP-43 had similar, but more severe, deficits. Through cross-rescue analysis, we demonstrated that FUS acted together with and downstream of TDP-43 in a common genetic pathway in neurons. Furthermore, we found that these proteins associated with each other in an RNA-dependent complex. Our results establish that FUS and TDP-43 function together in vivo and suggest that molecular pathways requiring the combined activities of both of these proteins may be disrupted in ALS and FTD.

Wang JW; Brent JR; Tomlinson A; Shneider NA; McCabe BD

2011-10-01

105

Loss of ALS-associated TDP-43 in zebrafish causes muscle degeneration, vascular dysfunction, and reduced motor neuron axon outgrowth.  

Science.gov (United States)

Mutations in the Tar DNA binding protein of 43 kDa (TDP-43; TARDBP) are associated with amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with TDP-43(+) inclusions (FTLD-TDP). To determine the physiological function of TDP-43, we knocked out zebrafish Tardbp and its paralogue Tardbp (TAR DNA binding protein-like), which lacks the glycine-rich domain where ALS- and FTLD-TDP-associated mutations cluster. tardbp mutants show no phenotype, a result of compensation by a unique splice variant of tardbpl that additionally contains a C-terminal elongation highly homologous to the glycine-rich domain of tardbp. Double-homozygous mutants of tardbp and tardbpl show muscle degeneration, strongly reduced blood circulation, mispatterning of vessels, impaired spinal motor neuron axon outgrowth, and early death. In double mutants the muscle-specific actin binding protein Filamin Ca is up-regulated. Strikingly, Filamin C is similarly increased in the frontal cortex of FTLD-TDP patients, suggesting aberrant expression in smooth muscle cells and TDP-43 loss-of-function as one underlying disease mechanism. PMID:23457265

Schmid, Bettina; Hruscha, Alexander; Hogl, Sebastian; Banzhaf-Strathmann, Julia; Strecker, Katrin; van der Zee, Julie; Teucke, Mathias; Eimer, Stefan; Hegermann, Jan; Kittelmann, Maike; Kremmer, Elisabeth; Cruts, Marc; Solchenberger, Barbara; Hasenkamp, Laura; van Bebber, Frauke; Van Broeckhoven, Christine; Edbauer, Dieter; Lichtenthaler, Stefan F; Haass, Christian

2013-03-01

106

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

UK PubMed Central (United Kingdom)

?-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.

Muńoz-Saez E; de Munck E; Arahuetes RM; Solas MT; Martínez AM; Miguel BG

2013-01-01

107

Divergent roles of ALS-linked proteins FUS/TLS and TDP-43 intersect in processing long pre-mRNAs.  

Science.gov (United States)

FUS/TLS (fused in sarcoma/translocated in liposarcoma) and TDP-43 are integrally involved in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. We found that FUS/TLS binds to RNAs from >5,500 genes in mouse and human brain, primarily through a GUGGU-binding motif. We identified a sawtooth-like binding pattern, consistent with co-transcriptional deposition of FUS/TLS. Depletion of FUS/TLS from the adult nervous system altered the levels or splicing of >950 mRNAs, most of which are distinct from RNAs dependent on TDP-43. Abundance of only 45 RNAs was reduced after depletion of either TDP-43 or FUS/TLS from mouse brain, but among these were mRNAs that were transcribed from genes with exceptionally long introns and that encode proteins that are essential for neuronal integrity. Expression levels of a subset of these were lowered after TDP-43 or FUS/TLS depletion in stem cell-derived human neurons and in TDP-43 aggregate-containing motor neurons in sporadic ALS, supporting a common loss-of-function pathway as one component underlying motor neuron death from misregulation of TDP-43 or FUS/TLS. PMID:23023293

Lagier-Tourenne, Clotilde; Polymenidou, Magdalini; Hutt, Kasey R; Vu, Anthony Q; Baughn, Michael; Huelga, Stephanie C; Clutario, Kevin M; Ling, Shuo-Chien; Liang, Tiffany Y; Mazur, Curt; Wancewicz, Edward; Kim, Aneeza S; Watt, Andy; Freier, Sue; Hicks, Geoffrey G; Donohue, John Paul; Shiue, Lily; Bennett, C Frank; Ravits, John; Cleveland, Don W; Yeo, Gene W

2012-09-30

108

Divergent roles of ALS-linked proteins FUS/TLS and TDP-43 intersect in processing long pre-mRNAs.  

UK PubMed Central (United Kingdom)

FUS/TLS (fused in sarcoma/translocated in liposarcoma) and TDP-43 are integrally involved in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. We found that FUS/TLS binds to RNAs from >5,500 genes in mouse and human brain, primarily through a GUGGU-binding motif. We identified a sawtooth-like binding pattern, consistent with co-transcriptional deposition of FUS/TLS. Depletion of FUS/TLS from the adult nervous system altered the levels or splicing of >950 mRNAs, most of which are distinct from RNAs dependent on TDP-43. Abundance of only 45 RNAs was reduced after depletion of either TDP-43 or FUS/TLS from mouse brain, but among these were mRNAs that were transcribed from genes with exceptionally long introns and that encode proteins that are essential for neuronal integrity. Expression levels of a subset of these were lowered after TDP-43 or FUS/TLS depletion in stem cell-derived human neurons and in TDP-43 aggregate-containing motor neurons in sporadic ALS, supporting a common loss-of-function pathway as one component underlying motor neuron death from misregulation of TDP-43 or FUS/TLS.

Lagier-Tourenne C; Polymenidou M; Hutt KR; Vu AQ; Baughn M; Huelga SC; Clutario KM; Ling SC; Liang TY; Mazur C; Wancewicz E; Kim AS; Watt A; Freier S; Hicks GG; Donohue JP; Shiue L; Bennett CF; Ravits J; Cleveland DW; Yeo GW

2012-11-01

109

TDP-43 high throughput screening analyses in neurodegeneration: Advantages and pitfalls.  

Science.gov (United States)

Dysfunctions in RNA processing and in particular the aberrant regulation of RNA binding proteins (RBPs) have recently been shown to play a fundamental role in the pathogenesis of neurodegenerative diseases. Understanding the pathogenic mechanisms involved will require the elucidation of the role(s) played by these RBPs in the general cell metabolism and neuronal survival in particular. In the past, the preferred approach has been to determine first of all the functional properties of the factor(s) of interest and then use this knowledge to determine targets in biologically relevant events. More recently, novel experimental approaches such as microarrays, RNA-seq and CLIP-seq have also become very popular to study RBPs. The advantage of these approaches, collectively known as high throughput screening (HTS), is their ability to determine gene expression changes or RNA/protein targets at a global cellular level. In theory, HTS strategies should be ideal for uncovering novel functional roles/targets of any RBP inside the cell. In practice, however, there are still difficulties in getting a coherent picture from all the huge amount of data they generate, frequently not validated experimentally and thus of unknown value. They may even act unfavorably towards a specific increase of knowledge of RBP functions, as the incomplete results are taken as solid data. In this work we will illustrate as an example the use of the HTS methodologies to characterize the interactions of a specific RBP: TDP-43. The multiple functions of this protein in RNA processing and its involvement in the pathogenesis of several forms of amyotrophic lateral sclerosis, frontotemporal lobar degeneration and other neurodegenerative diseases make it an excellent substrate for our analysis of the various advantages and limitations of different HTS experimental approaches. PMID:23500590

Buratti, Emanuele; Romano, Maurizio; Baralle, Francisco E

2013-03-14

110

TDP-43 high throughput screening analyses in neurodegeneration: Advantages and pitfalls.  

UK PubMed Central (United Kingdom)

Dysfunctions in RNA processing and in particular the aberrant regulation of RNA binding proteins (RBPs) have recently been shown to play a fundamental role in the pathogenesis of neurodegenerative diseases. Understanding the pathogenic mechanisms involved will require the elucidation of the role(s) played by these RBPs in the general cell metabolism and neuronal survival in particular. In the past, the preferred approach has been to determine first of all the functional properties of the factor(s) of interest and then use this knowledge to determine targets in biologically relevant events. More recently, novel experimental approaches such as microarrays, RNA-seq and CLIP-seq have also become very popular to study RBPs. The advantage of these approaches, collectively known as high throughput screening (HTS), is their ability to determine gene expression changes or RNA/protein targets at a global cellular level. In theory, HTS strategies should be ideal for uncovering novel functional roles/targets of any RBP inside the cell. In practice, however, there are still difficulties in getting a coherent picture from all the huge amount of data they generate, frequently not validated experimentally and thus of unknown value. They may even act unfavorably towards a specific increase of knowledge of RBP functions, as the incomplete results are taken as solid data. In this work we will illustrate as an example the use of the HTS methodologies to characterize the interactions of a specific RBP: TDP-43. The multiple functions of this protein in RNA processing and its involvement in the pathogenesis of several forms of amyotrophic lateral sclerosis, frontotemporal lobar degeneration and other neurodegenerative diseases make it an excellent substrate for our analysis of the various advantages and limitations of different HTS experimental approaches.

Buratti E; Romano M; Baralle FE

2013-09-01

111

Adenoviral expression of TDP-43 and FUS genes and shRNAs for protein degradation pathways in rodent motoneurons in vitro and in vivo.  

UK PubMed Central (United Kingdom)

Formation of cytoplasmic aggregates in neuronal and glial cells is one of the pathological hallmarks of amyotrophic lateral sclerosis (ALS). Mutations in two genes encoding transactivation response (TAR) DNA-binding protein 43 (TDP-43) and fused in sarcoma (FUS), both of which are main constituents of cytoplasmic aggregates, have been identified in patients with familial and sporadic ALS. Impairment of protein degradation machineries has also been recognized to participate in motoneuron degeneration in ALS. In the present study, we produced recombinant adenovirus vectors encoding wild type and mutant TDP-43 and FUS, and those encoding short hairpin RNAs (shRNAs) for proteasome (PSMC1), autophagy (ATG5), and endosome (VPS24) systems to investigate whether the coupled gene transductions in motoneurons by these adenoviruses elicit ALS pathology. Cultured neurons, astrocytes and oligodendrocytes differentiated from adult rat neural stem cells and motoneurons derived from mouse embryonic stem cells were successfully infected with these adenoviruses showing cytoplasmic aggregate formation. When these adenoviruses were injected into the facial nerves of adult rats, exogenous TDP-43 and FUS proteins were strongly expressed in facial motoneurons by a retrograde axonal transport of the adenoviruses. Co-infections of adenovirus encoding shRNA for PSMC1, ATG5 or VPS24 with TDP-43 or FUS adenovirus enhanced cytoplasmic aggregate formation in facial motoneurons, suggesting that impairment of protein degradation pathways accelerates formation of TDP-43 and FUS-positive aggregates in ALS.

Watabe K; Akiyama K; Kawakami E; Ishii T; Endo K; Yanagisawa H; Sango K; Tsukamoto M

2013-08-01

112

Downregulation of MicroRNA-9 in iPSC-Derived Neurons of FTD/ALS Patients with TDP-43 Mutations  

Science.gov (United States)

Transactive response DNA-binding protein 43 (TDP-43) is a major pathological protein in frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). There are many disease-associated mutations in TDP-43, and several cellular and animal models with ectopic overexpression of mutant TDP-43 have been established. Here we sought to study altered molecular events in FTD and ALS by using induced pluripotent stem cell (iPSC) derived patient neurons. We generated multiple iPSC lines from an FTD/ALS patient with the TARDBP A90V mutation and from an unaffected family member who lacked the mutation. After extensive characterization, two to three iPSC lines from each subject were selected, differentiated into postmitotic neurons, and screened for relevant cell-autonomous phenotypes. Patient-derived neurons were more sensitive than control neurons to 100 nM straurosporine but not to other inducers of cellular stress. Three disease-relevant cellular phenotypes were revealed under staurosporine-induced stress. First, TDP-43 was localized in the cytoplasm of a higher percentage of patient neurons than control neurons. Second, the total TDP-43 level was lower in patient neurons with the A90V mutation. Third, the levels of microRNA-9 (miR-9) and its precursor pri-miR-9-2 decreased in patient neurons but not in control neurons. The latter is likely because of reduced TDP-43, as shRNA-mediated TDP-43 knockdown in rodent primary neurons also decreased the pri-miR-9-2 level. The reduction in miR-9 expression was confirmed in human neurons derived from iPSC lines containing the more pathogenic TARDBP M337V mutation, suggesting miR-9 downregulation might be a common pathogenic event in FTD/ALS. These results show that iPSC models of FTD/ALS are useful for revealing stress-dependent cellular defects of human patient neurons containing rare TDP-43 mutations in their native genetic contexts.

Zhang, Zhijun; Almeida, Sandra; Lu, Yubing; Nishimura, Agnes L.; Peng, Lingtao; Sun, Danqiong; Wu, Bei; Karydas, Anna M.; Tartaglia, Maria C.; Fong, Jamie C.; Miller, Bruce L.; Farese, Robert V.; Moore, Melissa J.; Shaw, Christopher E.; Gao, Fen-Biao

2013-01-01

113

Calcium channel agonists protect against neuromuscular dysfunction in a genetic model of TDP-43 mutation in ALS.  

UK PubMed Central (United Kingdom)

TAR DNA binding protein (TDP-43, encoded by the TARDBP gene) has recently been shown to be associated with amyotrophic lateral sclerosis (ALS), but the early pathophysiological deficits causing impairment in motor function are unknown. Here we expressed the wild-type human gene (wtTARDBP) or the ALS mutation G348C (mutTARDBP) in zebrafish larvae and characterized their motor (swimming) activity and the structure and function of their neuromuscular junctions (NMJs). Of these groups only mutTARDBP larvae showed impaired swimming and increased motoneuron vulnerability with reduced synaptic fidelity, reduced quantal transmission, and more orphaned presynaptic and postsynaptic structures at the NMJ. Remarkably, all behavioral and cellular features were stabilized by chronic treatment with either of the L-type calcium channel agonists FPL 64176 or Bay K 8644. These results indicate that expression of mutTARDBP results in defective NMJs and that calcium channel agonists could be novel therapeutics for ALS.

Armstrong GA; Drapeau P

2013-01-01

114

TDP-43 pathology in a patient carrying G2019S LRRK2 mutation and a novel p.Q124E MAPT.  

Science.gov (United States)

Leucine-rich repeat kinase 2 (LRRK2) mutation is the most common cause of genetic-related parkinsonism and is usually associated with Lewy body pathology; however, tau, ?-synuclein, and ubiquitin pathologies have also been reported. We report the case of a patient carrying the LRRK2 G2019S mutation and a novel heterozygous variant c.370C>G, p.Q124E in exon 4 of the microtubule-associated protein tau (MAPT). The patient developed parkinsonism with good levodopa response in her 70s. Neuropathological analysis revealed nigral degeneration and Alzheimer-type tau pathology without Lewy bodies. Immunohistochemical staining using phospho-TDP-43 antibodies identified occasional TDP-43 pathology in the hippocampus, temporal neocortex, striatum, and substantia nigra. However, TDP-43 pathology was not identified in another 4 archival LRRK2 G2019S cases with Lewy body pathology available in the Queen Square Brain Bank. Among other published cases of patients carrying LRRK2 G2019S mutation, only 3 were reportedly evaluated for TDP-43 pathology, and the results were negative. The role of the MAPT variant in the clinical and pathological manifestation in LRRK2 cases remains to be determined. PMID:23664753

Ling, Helen; Kara, Eleanna; Bandopadhyay, Rina; Hardy, John; Holton, Janice; Xiromerisiou, Georgia; Lees, Andrew; Houlden, Henry; Revesz, Tamas

2013-05-09

115

TDP-43 pathology in a patient carrying G2019S LRRK2 mutation and a novel p.Q124E MAPT.  

UK PubMed Central (United Kingdom)

Leucine-rich repeat kinase 2 (LRRK2) mutation is the most common cause of genetic-related parkinsonism and is usually associated with Lewy body pathology; however, tau, ?-synuclein, and ubiquitin pathologies have also been reported. We report the case of a patient carrying the LRRK2 G2019S mutation and a novel heterozygous variant c.370C>G, p.Q124E in exon 4 of the microtubule-associated protein tau (MAPT). The patient developed parkinsonism with good levodopa response in her 70s. Neuropathological analysis revealed nigral degeneration and Alzheimer-type tau pathology without Lewy bodies. Immunohistochemical staining using phospho-TDP-43 antibodies identified occasional TDP-43 pathology in the hippocampus, temporal neocortex, striatum, and substantia nigra. However, TDP-43 pathology was not identified in another 4 archival LRRK2 G2019S cases with Lewy body pathology available in the Queen Square Brain Bank. Among other published cases of patients carrying LRRK2 G2019S mutation, only 3 were reportedly evaluated for TDP-43 pathology, and the results were negative. The role of the MAPT variant in the clinical and pathological manifestation in LRRK2 cases remains to be determined.

Ling H; Kara E; Bandopadhyay R; Hardy J; Holton J; Xiromerisiou G; Lees A; Houlden H; Revesz T

2013-12-01

116

Endogenous progesterone levels and frontotemporal dementia: modulation of TDP-43 and Tau levels in vitro and treatment of the A315T TARDBP mouse model.  

Science.gov (United States)

Frontotemporal dementia (FTD) is associated with motor neurone disease (FTD-MND), corticobasal syndrome (CBS) and progressive supranuclear palsy syndrome (PSPS). Together, this group of disorders constitutes a major cause of young-onset dementia. One of the three clinical variants of FTD is progressive nonfluent aphasia (PNFA), which is focused on in this study. The steroid hormone progesterone (PROG) is known to have an important role as a neurosteroid with potent neuroprotective and promyelination properties. In a case-control study of serum samples (39 FTD, 91 controls), low serum PROG was associated with FTD overall. In subgroup analysis, low PROG levels were significantly associated with FTD-MND and CBS, but not with PSPS or PNFA. PROG levels of >195 pg/ml were significantly correlated with lower disease severity (frontotemporal dementia rating scale) for individuals with CBS. In the human neuroblastoma SK-N-MC cell line, exogenous PROG (9300-93,000 pg/ml) had a significant effect on overall Tau and nuclear TDP-43 levels, reducing total Tau levels by ?1.5-fold and increasing nuclear TDP-43 by 1.7- to 2.0-fold. Finally, elevation of plasma PROG to a mean concentration of 5870 pg/ml in an Ala315Thr (A315T) TARDBP transgenic mouse model significantly reduced the rate of loss of locomotor control in PROG-treated, compared with placebo, mice. The PROG treatment did not significantly increase survival of the mice, which might be due to the limitation of the transgenic mouse to accurately model TDP-43-mediated neurodegeneration. Together, our clinical, cellular and animal data provide strong evidence that PROG could be a valid therapy for specific related disorders of FTD. PMID:23798570

Dang, Theresa N T; Dobson-Stone, Carol; Glaros, Elias N; Kim, Woojin S; Hallupp, Marianne; Bartley, Lauren; Piguet, Olivier; Hodges, John R; Halliday, Glenda M; Double, Kay L; Schofield, Peter R; Crouch, Peter J; Kwok, John B J

2013-06-20

117

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; Kouri Naomi; Baker Matt; Finch NiCole; DeJesus-Hernandez Mariely; Gonzalez John; Chidamparam Kumaravel; Josephs Keith A; Boeve Bradley F; Graff-Radford Neill R; Crook Julia; Dickson Dennis W; Rademakers Rosa

2011-01-01

118

Intra-familial clinical heterogeneity due to FTLD-U with TDP-43 proteinopathy caused by a novel deletion in progranulin gene (PGRN).  

UK PubMed Central (United Kingdom)

Frontotemporal dementia (FTD) is one of the commonest forms of early-onset dementia, accounting for up to 20% of all dementia patients. Recently, it has been shown that mutations in progranulin gene (PGRN) cause many familial cases of FTD. Members of a family affected by FTD spectrum disorders were ascertained in Poland and Canada. Clinical, radiological, molecular, genetic, and pathological studies were performed. A sequencing analysis of PGRN exons 1-13 was performed in the proband. Genotyping of the identified PGRN mutation and pathological analysis was carried out in the proband's brother. The onset of symptoms of FTD in the proband included bradykinesia, apathy, and somnolence followed by changes in personality, cognitive deficits, and psychotic features. The proband's clinical diagnosis was FTD and parkinsonism (FTDP). DNA sequence analysis of PGRN revealed a novel, heterozygous mutation in exon 11 (g.2988_2989delCA, P439_R440fsX6). The mutation introduced a premature stop codon at position 444. The proband's brother with the same mutation had a different course first presenting as progressive non-fluent aphasia, and later evolving symptoms of behavioral variant of FTD. He also developed parkinsonism late in the disease course evolving into corticobasal syndrome. Pathological analysis in the brother revealed Frontotemporal Lobar Degeneration-Ubiquitin (FTLD-U)/TDP-43 positive pathology. The novel PGRN mutation is a disease-causing mutation and is associated with substantial intra-familial clinical heterogeneity. Although presenting features were different, rapid and substantial deterioration in the disease course was observed in both family members.

Gabryelewicz T; Masellis M; Berdynski M; Bilbao JM; Rogaeva E; St George-Hyslop P; Barczak A; Czyzewski K; Barcikowska M; Wszolek Z; Black SE; Zekanowski C

2010-01-01

119

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  

Directory of Open Access Journals (Sweden)

Full Text Available 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 frontotemporal dementia, associated with either clinically possible or definite MND. Neuropathological examination revealed hallmarks of FTLD-TDP with major involvement of subcortical and, in particular, mesencephalic structures. These cases differed in onset and progression of clinical manifestations as well as distribution of histopathological changes in the brain and spinal cord. Two cases were sporadic, whereas the third case had a pathological variation in the progranulin gene 102 delC. Conclusions Association of a "progressive supranuclear palsy-like" syndrome with marked visuospatial impairment, motor neuron disease and early behavioral disturbances may represent a clinically distinct phenotype of FTLD-TDP. Our observations further support the concept that TDP-43 proteinopathies represent a spectrum of disorders, where preferential localization of pathogenetic inclusions and neuronal cell loss defines clinical phenotypes ranging from frontotemporal dementia with or without motor neuron disease, to corticobasal syndrome and to a progressive supranuclear palsy-like syndrome.

Rusina Robert; Kovacs Gabor G; Fiala Jind?ich; Hort Jakub; Ridzo? Petr; Holmerová Iva; Ströbel Thomas; Mat?j Radoslav

2011-01-01

120

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

Directory of Open Access Journals (Sweden)

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; Gendron Tania F; Xu Ya-Fei; Ko Li-Wen; Yen Shu-Hui; Petrucelli Leonard

2010-01-01

 
 
 
 
121

TDP-43 frontotemporal lobar degeneration and autoimmune disease.  

UK PubMed Central (United Kingdom)

BACKGROUND: The aetiology and pathogenesis of non-genetic forms of frontotemporal dementia (FTD) is unknown and even with the genetic forms of FTD, pathogenesis remains elusive. Given the association between systemic inflammation and other neurodegenerative processes, links between autoimmunity and FTD need to be explored. OBJECTIVE: To describe the prevalence of systemic autoimmune disease in semantic variant primary progressive aphasia (svPPA), a clinical cohort, and in progranulin (PGRN) mutation carriers compared with neurologically healthy normal controls (NC) and Alzheimer's disease (AD) as dementia controls. DESIGN: Case control. SETTING: Academic medical centres. PARTICIPANTS: 129 svPPA, 39 PGRN, 186 NC and 158 AD patients underwent chart review for autoimmune conditions. A large subset of svPPA, PGRN and NC cohorts underwent serum analysis for tumour necrosis factor ? (TNF-?) levels. OUTCOME MEASURES: ?(2) Comparison of autoimmune prevalence and follow-up logistic regression. RESULTS: There was a significantly increased risk of autoimmune disorders clustered around inflammatory arthritides, cutaneous disorders and gastrointestinal conditions in the svPPA and PGRN cohorts. Elevated TNF-? levels were observed in svPPA and PGRN compared with NC. CONCLUSIONS: svPPA and PGRN are associated with increased prevalence of specific and related autoimmune diseases compared with NC and AD. These findings suggest a unique pattern of systemic inflammation in svPPA and PGRN and open new research avenues for understanding and treating disorders associated with underlying transactive response DNA-binding protein 43 aggregation.

Miller ZA; Rankin KP; Graff-Radford NR; Takada LT; Sturm VE; Cleveland CM; Criswell LA; Jaeger PA; Stan T; Heggeli KA; Hsu SC; Karydas A; Khan BK; Grinberg LT; Gorno-Tempini ML; Boxer AL; Rosen HJ; Kramer JH; Coppola G; Geschwind DH; Rademakers R; Seeley WW; Wyss-Coray T; Miller BL

2013-09-01

122

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

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 PrPS...

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

123

Circulating and tissue endothelin immunoreactivity in hypercholesterolemic pigs.  

UK PubMed Central (United Kingdom)

BACKGROUND: Hypercholesterolemia is characterized by a coronary vasoconstrictive response to the endothelium-dependent vasodilator acetylcholine. This abnormality may be due to reduced synthesis of endothelium-derived relaxing factor and/or enhanced synthesis and release of an endothelium-derived contracting factor. Endothelin is an endothelium-derived vasoconstrictor and mitogenic peptide that is present in normal plasma, and its circulating concentrations are elevated in disease states that are characterized by abnormal endothelium-dependent relaxation to acetylcholine. The current studies were designed to test the hypotheses that experimental hypercholesterolemia results in elevation of plasma and tissue endothelin immunoreactivity and that the abnormal acetylcholine-evoked coronary vasoconstriction in the hypercholesterolemic animals is associated with further elevation of plasma endothelin. METHODS AND RESULTS: Plasma concentrations and molecular forms of endothelin immunoreactivity were determined following 2% cholesterol diet for 4 months in pigs and during intracoronary acetylcholine administration. Second, we assessed the presence of endothelin in the coronary vascular wall by using immunohistochemistry. Hypercholesterolemia elevated plasma endothelin concentration and enhanced coronary artery tissue endothelin immunoreactivity. The endothelium-dependent vasodilator acetylcholine further increases plasma endothelin in hypercholesterolemia in association with coronary vasoconstriction. The predominant molecular form of endothelin in hypercholesterolemia is the biological active endothelin-1. CONCLUSIONS: This study suggests a role for endothelin as an early participant and a marker for the endothelial dysfunction in hypercholesterolemia as well as a participant in the atherogenic process.

Lerman A; Webster MW; Chesebro JH; Edwards WD; Wei CM; Fuster V; Burnett JC Jr

1993-12-01

124

Antibodies immunoreactive with mutant hydroxypenylpyruvatedioxygenase  

UK PubMed Central (United Kingdom)

Antibodies immunoreactive to mutant Pseudomonas HPPD are provided, and in an embodiment the mutant HPPD is one in which the wild-type HPPD is substituted at residue 336 with tryptophan for glycine. Also provided are hybridomas producing the antibodies, as well as methods of making and using the antibodies.

SEKAR VAITHILINGAM; HELD BRUCE; CHUNG KYU; RUSSELL JR PAUL F

125

[Parkinson disease and amyotrophic lateral sclerosis. Tauopathies, TDP-43 and SOD mutations].  

Science.gov (United States)

In addition to a large number of clinical descriptions of atypical cases, recent pathological, biochemical and genetic studies challenge the view that amyotrophic lateral sclerosis (ALS) is a disorder restricted to the pyramidal motor system. Relations between ALS, Parkinson disease, fronto-temporal dementia, progressive supranuclear paralysis, and cortico-basal degeneration have now been identified. We propose a review of the topic and discuss the contribution of various clinical and pathological features leading to consider motoneuron diseases as neurodegenerative processes included in a broad spectrum of tauopathies. PMID:18808763

Le Forestier, N; Lacomblez, L; Meininger, V

2008-06-24

126

[Parkinson disease and amyotrophic lateral sclerosis. Tauopathies, TDP-43 and SOD mutations].  

UK PubMed Central (United Kingdom)

In addition to a large number of clinical descriptions of atypical cases, recent pathological, biochemical and genetic studies challenge the view that amyotrophic lateral sclerosis (ALS) is a disorder restricted to the pyramidal motor system. Relations between ALS, Parkinson disease, fronto-temporal dementia, progressive supranuclear paralysis, and cortico-basal degeneration have now been identified. We propose a review of the topic and discuss the contribution of various clinical and pathological features leading to consider motoneuron diseases as neurodegenerative processes included in a broad spectrum of tauopathies.

Le Forestier N; Lacomblez L; Meininger V

2009-01-01

127

TDP-43 pathology in familial frontotemporal dementia and motor neuron disease without Progranulin mutations  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Frontotemporal dementia is accompanied by motor neuron disease (FTD + MND) in ?10% of cases. There is accumulating evidence for a clinicopathological overlap between FTD and MND based on observations of familial aggregation and neuropathological findings of ubiquitin-positive neuronal cytoplasmatic ...

Seelaar, H.; Jurgen Schelhaas, H.; Azmani, A.; Küsters, B.; Rosso, S.M.; Majoor-Krakauer, D.F.; Rijik, M.C. de; Rizzu, P.

128

Loss of TDP-43 causes age-dependent progressive motor neuron degeneration.  

UK PubMed Central (United Kingdom)

Amyotrophic lateral sclerosis is a devastating, progressive neurodegenerative disease that affects upper and lower motor neurons. Although several genes are identified as the cause of familial cases, the pathogeneses of sporadic forms, which account for 90% of amyotrophic lateral sclerosis, have not been elucidated. Transactive response DNA-binding protein 43 a nuclear protein regulating RNA processing, redistributes to the cytoplasm and forms aggregates, which are the histopathological hallmark of sporadic amyotrophic lateral sclerosis, in affected motor neurons, suggesting that loss-of-function of transactive response DNA-binding protein 43 is one of the causes of the neurodegeneration. To test this hypothesis, we assessed the effects of knockout of transactive response DNA-binding protein 43 in mouse postnatal motor neurons using Cre/loxp system. These mice developed progressive weight loss and motor impairment around the age of 60 weeks, and exhibited degeneration of large motor axon, grouped atrophy of the skeletal muscle, and denervation in the neuromuscular junction. The spinal motor neurons lacking transactive response DNA-binding protein 43 were not affected for 1 year, but exhibited atrophy at the age of 100 weeks; whereas, extraocular motor neurons, that are essentially resistant in amyotrophic lateral sclerosis, remained preserved even at the age of 100 weeks. Additionally, ultra structural analysis revealed autolysosomes and autophagosomes in the cell bodies and axons of motor neurons of the 100-week-old knockout mice. In summary, the mice in which transactive response DNA-binding protein 43 was knocked-out specifically in postnatal motor neurons exhibited an age-dependent progressive motor dysfunction accompanied by neuropathological alterations, which are common to sporadic amyotrophic lateral sclerosis. These findings suggest that transactive response DNA-binding protein 43 plays an essential role in the long term maintenance of motor neurons and that loss-of-function of this protein seems to contribute to the pathogenesis of amyotrophic lateral sclerosis.

Iguchi Y; Katsuno M; Niwa J; Takagi S; Ishigaki S; Ikenaka K; Kawai K; Watanabe H; Yamanaka K; Takahashi R; Misawa H; Sasaki S; Tanaka F; Sobue G

2013-05-01

129

Nail abnormalities  

Science.gov (United States)

Beau's lines; Fingernail abnormalities; Spoon nails; Onycholysis; Leukonychia; Koilonychia; Brittle nails ... lines can occur after illness, injury to the nail, and when you are malnourished . Brittle nails are ...

130

Meiotic abnormalities  

Energy Technology Data Exchange (ETDEWEB)

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

131

Increased Ki-67 immunoreactivity in the white matter in hemimegalencephaly.  

UK PubMed Central (United Kingdom)

Hemimegalencephaly (HMG) is a developmental brain disorder characterized by an enlarged unilateral hemisphere with cortical malformation comprising abnormal hypertrophic cells. To address the proliferative status of HMG, Ki-67 immunoreactivity was investigated in HMG specimens obtained during epilepsy surgery. Nine HMG tissues were stained with a Ki-67 antibody and Ki-67 labeling index in the malformed cortex, and the underlying white matter was measured separately and compared with tissues from focal cortical dysplasias and normal brains from autopsy. In HMG tissues, Ki-67-positive cells were scattered in both the gray and white matter, with a significantly higher Ki-67 labeling index in the white matter compared with gray matter. No dysmorphic neuron or balloon cell was stained for Ki-67. As Ki-67 immunoreactivity overlapped with that of ionized calcium-binding adaptor protein-1, Ki-67-positive cells were identified as microglia. In HMG, microglia were activated and entered into a proliferative status with higher distribution in the white matter, implying an ongoing neuroinflammatory process involving the white matter.

Munakata M; Watanabe M; Otsuki T; Itoh M; Uematsu M; Saito Y; Honda R; Kure S

2013-08-01

132

Chromosomal abnormalities  

International Nuclear Information System (INIS)

Cytogenetic studies from the peripheral blood of a patient with malignant lymphoma and rhematoid arthritis who was treated with intra-articular gold Au 198 revealed mosaicism with a normal female metaphase and a 43-chromosome metaphase. The abnormal cell line showed six missing normal chromosomes and three morphologically abnormal chromosomes. The trypsin-digested G-banding metaphases showed that the marker chromosomes were an isochromosome of the long arm of chromosome 17, a translocated chromosome that involved the long arm of chromosome 4 and a chromosome 16, and a translocated chromosome that involved the long arm of chromosome 4 and a chromosome 5. It is tempting to conclude that these abnormalities were due to the gold Au 198 treatment, but we cannot exclude other possibilities.

1980-01-01

133

ANTIBODIES IMMUNOREACTIVE WITH MUTANT HYDROXYPENYLPYRUVATE DIOXYGENASE  

UK PubMed Central (United Kingdom)

Antibodies immunoreactive to mutant Pseudomonas HPPD are provided, and in an embodiment the mutant HPPD is one in which the wild-type HPPD is substituted at residue 336 with tryptophan for glycine. Also provided are hybridomas producing the antibodies, as well as methods of making and using the antibodies.

SEKAR VAITHILINGAM; HELD BRUCE; CHUNG KYU; RUSSELL JR PAUL

134

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

CERN Multimedia

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

135

Canine and feline pancreatic lipase immunoreactivity.  

UK PubMed Central (United Kingdom)

The diagnosis of pancreatitis in dogs and cats can be challenging. Several diagnostic tests have been evaluated over the years, but the majority have been shown to be of limited utility owing to poor performance or limited availability or because invasive procedures are required. Assays for the measurement of pancreatic lipase immunoreactivity (cPLI for dogs and fPLI for cats) were first developed over a decade ago and now include Spec cPL and SNAP cPL for dogs and Spec fPL and SNAP fPL for cats. Owing to their high sensitivity and specificity for pancreatitis compared with those of other serum tests, concentrations of cPLI and fPLI have been demonstrated to be the serum tests of choice for evaluation of dogs and cats, respectively, suspected of having pancreatitis. False-positive and false-negative results can occur, and recognition of the limitations of pancreatic lipase immunoreactivity assays is important. As there is currently no gold standard for antemortem diagnosis of pancreatitis in dogs and cats, the combination of a complete history and physical examination, measurement of pancreatic lipase immunoreactivity, and ultrasonographic examination of the pancreas is the best approach for an accurate noninvasive diagnosis of pancreatitis.

Xenoulis PG; Steiner JM

2012-09-01

136

High affinity choline transporter immunoreactivity in rat ileum myenteric nerves.  

Science.gov (United States)

Recently, an antibody against the choline transporter (CHT), an essential molecule involved in ACh uptake, was used to label cholinergic nerves in the central nervous system; however, the enteric nervous system (ENS) was not examined. The present study localised CHT immunoreactivity (CHT-IR) within the rat ileum ENS and determined whether it colocalised with immunoreactivity for markers of cholinergic, tachykinergic and nitrergic circuitry. Segments of rat ileum were fixed, prepared for sectioning or whole-mounts and incubated with anti-CHT antisera followed by a fluorescent secondary antibody. Samples were double-labelled with antibodies to nitric oxide synthase, substance P (SP), common choline acetyltransferase (cChAT) and vesicular acetylcholine transporter (VAChT). CHT-IR was present in varicosities of nerve fibres in the myenteric plexus and muscle layers of rat ileum. In the myenteric ganglia, CHT-IR was found in nerve fibres and the cytoplasm of some nerve cell bodies. In the myenteric ganglia, no CHT/cChAT-immunoreactive neurons were present. A small number of CHT/SP-immunoreactive neurons and CHT/SP-immunoreactive nerve fibres clustered around unlabelled neurons. CHT-IR colocalised with VAChT-IR in the myenteric plexus but only half of the CHT-immunoreactive myenteric nerve fibres were VAChT-immunoreactive and half of VAChT-immunoreactive fibres were CHT-immunoreactive. In the circular muscle, 75% of CHT-immunoreactive fibres were VAChT-immunoreactive. Thus, the anti-CHT antiserum labels neurons and nerve fibres in the rat ENS. It does not label cholinergic cChAT-immunoreactive neurons, although it does immunostain cholinergic VAChT-immunoreactive nerve fibres and a population of nerves that are not VAChT-immunoreactive. PMID:17093920

Harrington, Andrea M; Hutson, J M; Southwell, Bridget R

2006-11-09

137

Immunoreactive trypsin and neonatalscreening for cystic fibrosis  

International Nuclear Information System (INIS)

[en] 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[fr] La trypsine immunoreactive a ete mesuree, dans le sang seche sur papier filtre, chez 160.822 nouveau-nes de 5 jours dans le cadre d'un programme regional de depistage neonatal systematique de la mucoviscidose. Un second test a ete realise chez 492 nouveau-nes pour lesquels le premier resultat etait superieur a 900 ?g/l; l'elevation du taux de trypsine a ete confirmee dans 55 cas. Les tests de la sueur ont confirme la mucoviscidose chez 43 nourrissons, mais ils etaient normaux dans 12 cas. Au cours de cette etude, 51 cas de mucoviscidose ont ete reperes au total: 43 par le depistage, 6 par la presence d'un ileus meconial; le diagnostic precoce a donc ete obtenu dans 49 cas sur 51. Deux nouveau-nes qui avaient un taux de trypsine immunoreactive strictement normal n'ont pu etre identifies par le test de depistage

1988-01-01

138

PACAP immunoreactivity in human malignant tumor samples and cardiac diseases.  

UK PubMed Central (United Kingdom)

Pituitary adenylate cyclase activating polypeptide (PACAP) is a pleiotropic and multifunctional neuropeptide having important roles in various physiological processes. Recent trends in PACAP research point to the clinical introduction of PACAP or its analogs/fragments possibly in the near future. Recently, we have shown the presence of PACAP in human plasma, milk, placenta, and follicular fluid samples. However, relatively few data are available on PACAP in human tissues from patients with different disorders. The aim of the present study was to determine, by radioimmunoassay, the tissue level of PACAP38-like immunoreactivity (LI) and PACAP27-LI in different primary non-small cell lung cancer, colon tumor samples, and in cardiac muscle samples from patients suffering from ischemic heart disease and valvular disorders. We also labeled the PAC1 receptors in human cardiac cells. All samples showed significantly higher PACAP38-LI compared with PACAP27-LI. We found significantly lower levels of PACAP38-LI and PACAP27-LI in tumoral and peripheral samples compared with normal healthy tissue in both lung and colon cancers. Further investigations are necessary to describe the exact function of PACAP in oncogenesis. We showed that PACAP38-LI and PACAP27-LI are significantly higher in ischemic heart diseases compared with valvular abnormalities, suggesting that PACAP might play a role in ischemic heart disorders.

Szanto Z; Sarszegi Z; Reglodi D; Nemeth J; Szabadfi K; Kiss P; Varga A; Banki E; Csanaky K; Gaszner B; Pinter O; Szalai Z; Tamas A

2012-11-01

139

PACAP immunoreactivity in human malignant tumor samples and cardiac diseases.  

Science.gov (United States)

Pituitary adenylate cyclase activating polypeptide (PACAP) is a pleiotropic and multifunctional neuropeptide having important roles in various physiological processes. Recent trends in PACAP research point to the clinical introduction of PACAP or its analogs/fragments possibly in the near future. Recently, we have shown the presence of PACAP in human plasma, milk, placenta, and follicular fluid samples. However, relatively few data are available on PACAP in human tissues from patients with different disorders. The aim of the present study was to determine, by radioimmunoassay, the tissue level of PACAP38-like immunoreactivity (LI) and PACAP27-LI in different primary non-small cell lung cancer, colon tumor samples, and in cardiac muscle samples from patients suffering from ischemic heart disease and valvular disorders. We also labeled the PAC1 receptors in human cardiac cells. All samples showed significantly higher PACAP38-LI compared with PACAP27-LI. We found significantly lower levels of PACAP38-LI and PACAP27-LI in tumoral and peripheral samples compared with normal healthy tissue in both lung and colon cancers. Further investigations are necessary to describe the exact function of PACAP in oncogenesis. We showed that PACAP38-LI and PACAP27-LI are significantly higher in ischemic heart diseases compared with valvular abnormalities, suggesting that PACAP might play a role in ischemic heart disorders. PMID:22648511

Szanto, Z; Sarszegi, Zs; Reglodi, D; Nemeth, J; Szabadfi, K; Kiss, P; Varga, A; Banki, E; Csanaky, K; Gaszner, B; Pinter, O; Szalai, Zs; Tamas, A

2012-05-31

140

Immunoreactivity and expression of amylin in gastroenteropancreatic endocrine tumors.  

UK PubMed Central (United Kingdom)

Amylin was isolated from human insulinomas, but there has been only preliminary data regarding whether this peptide can also be detected in other types of gastroenteropancreatic endocrine tumors. In the present study, immunohistochemical staining of 87 gastroenteropancreatic endocrine tumors demonstrated amylin immunoreactivity in 21.8% of the neoplasmas. Thirteen of 15 insulinomas, three of 21 gastrinomas, two of 29 nonfunctioning tumors, and one of 18 carcinoids were amylin-immunoreactive. Seventeen of the 19 amylin-immunoreactive tumors were primarily located in the pancreas, but two tumors were found in the intestine. Measurements of amylin messenger RNA expression in a few tumors revealed amylin synthesis in these tumors. Amylin immunoreactivity did not correlate with invasion and metastasis. However, the rate of curative resections was significantly higher in amylin-immunoreactive tumors. These results demonstrate for the first time that amylin immunoreactivity is not restricted to insulinomas and can also occur rarely in endocrine tumors of the intestine.

Eissele R; Neuhaus C; Trautmann ME; Funk A; Arnold R; Höfler H

1993-07-01

 
 
 
 
141

Immunoreactivity and expression of amylin in gastroenteropancreatic endocrine tumors.  

Science.gov (United States)

Amylin was isolated from human insulinomas, but there has been only preliminary data regarding whether this peptide can also be detected in other types of gastroenteropancreatic endocrine tumors. In the present study, immunohistochemical staining of 87 gastroenteropancreatic endocrine tumors demonstrated amylin immunoreactivity in 21.8% of the neoplasmas. Thirteen of 15 insulinomas, three of 21 gastrinomas, two of 29 nonfunctioning tumors, and one of 18 carcinoids were amylin-immunoreactive. Seventeen of the 19 amylin-immunoreactive tumors were primarily located in the pancreas, but two tumors were found in the intestine. Measurements of amylin messenger RNA expression in a few tumors revealed amylin synthesis in these tumors. Amylin immunoreactivity did not correlate with invasion and metastasis. However, the rate of curative resections was significantly higher in amylin-immunoreactive tumors. These results demonstrate for the first time that amylin immunoreactivity is not restricted to insulinomas and can also occur rarely in endocrine tumors of the intestine. PMID:8317551

Eissele, R; Neuhaus, C; Trautmann, M E; Funk, A; Arnold, R; Höfler, H

1993-07-01

142

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 cere...

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

143

Progressive nonfluent aphasia: a rare clinical subtype of FTLD-TDP in Japan.  

UK PubMed Central (United Kingdom)

Progressive nonfluent aphasia (PNFA) is a clinical subtype of frontotemporal lobar degeneration (FTLD). FTLD with tau accumulation (FTLD-tau) and FTLD with TDP-43 accumulation (FTLD-TDP) both cause PNFA. We reviewed clinical records of 29 FTLD-TDP cases in the brain archive of our institute and found only one case of PNFA. The patient was an 81-year-old male at death. There was no family history of dementia or aphasia. He presented with slow, labored and nonfluent speech at age 75. Behavioral abnormality and movement disorders were absent. MRI at age 76 demonstrated atrophy of the perisylvian regions, including the inferior frontal gyrus, insular gyrus and superior temporal gyrus. The atrophy was more severe in the left hemisphere than the right. On post mortem examinations, neuronal loss was evident in these regions as well as in the substantia nigra. There were abundant TDP-43-immunoreactive neuronal cytoplasmic inclusions and round or irregular-shaped structures in the affected cerebral cortices. A few dystrophic neurites and neuronal intranuclear inclusions were also seen. FTLD-TDP showing PNFA seems to be rare but does exist in Japan, similar to that in other countries.

Aoki N; Tsuchiya K; Kobayashi Z; Arai T; Togo T; Miyazaki H; Kondo H; Ishizu H; Uchikado H; Katsuse O; Hirayasu Y; Akiyama H

2012-06-01

144

Distribution of parvalbumin immunoreactivity in the vertebrate retina.  

UK PubMed Central (United Kingdom)

Parvalbumin, a calcium-binding protein thought to buffer intracellular calcium, is expressed in selected neuronal and non-neuronal cell populations. We used a well-characterized antibody directed against parvalbumin to investigate the distribution of parvalbumin in the retina of twelve vertebrate species to evaluate patterns of cellular expression for recurrent functional features. Parvalbumin immunoreactivity was displayed by subpopulations of ganglion, amacrine, bipolar and horizontal cells in different species-specific combinations. In the pigeon retina, subpopulations of amacrine, ganglion and bipolar cells were immunoreactive for parvalbumin. Parvalbumin immunoreactive bipolar cells in this species were mostly confined to the temporal dorsal region of the retina. In the owl, no immunoreactive amacrine cells were found, but many bipolar cells displayed parvalbumin immunoreactivity. In the teleost retina, amacrine and ganglion cells were found to be immunoreactive for parvalbumin. A high degree of species-specific variation was encountered in the mammalian retina. The most consistent finding within this class was that subpopulations of parvalbumin-immunoreactive amacrine cells were consistently observed in every species. In the rabbit, horizontal and ganglion cells displaying parvalbumin immunoreactivity were also seen. In rodents (hamster, ground squirrel), parvalbumin immunoreactivity was displayed by subpopulations of amacrine cells and, in the squirrel, by some ganglion cells as well. In the cat and in the baboon retina, parvalbumin immunoreactivity was found in horizontal cells, ganglion cells and a subpopulation of amacrine cells. The distribution of parvalbumin immunoreactive neurons in the vertebrate retinae studied showed no systematic correlation with phylogenetic proximity. The expression of parvalbumin within the systems of retinal neurons may therefore reflect the functional needs of different visual behaviors.

Sanna PP; Keyser KT; Celio MR; Karten HJ; Bloom FE

1993-01-01

145

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

146

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

147

Mapping of neurokinin-like immunoreactivity in the human brainstem  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Using an indirect immunoperoxidase technique, we have studied the distribution of immunoreactive fibers and cell bodies containing neurokinin in the adult human brainstem with no prior history of neurological or psychiatric disease. Results Clusters of immunoreactive cell bodies and high densities of neurokinin-immunoreactive fibers were located in the periaqueductal gray, the dorsal motor nucleus of the vagus and in the reticular formation of the medulla, pons and mesencephalon. Moreover, immunoreactive cell bodies were found in the inferior colliculus, the raphe obscurus, the nucleus prepositus hypoglossi, and in the midline of the anterior medulla oblongata. In general, immunoreactive fibers containing neurokinin were observed throughout the whole brainstem. In addition to the nuclei mentioned above, the highest densities of such immunoreactive fibers were located in the spinal trigeminal nucleus, the lateral reticular nucleus, the nucleus of the solitary tract, the superior colliculus, the substantia nigra, the nucleus ambiguus, the gracile nucleus, the cuneate nucleus, the motor hypoglossal nucleus, the medial and superior vestibular nuclei, the nucleus prepositus hypoglossi and the interpeduncular nucleus. Conclusion The widespread distribution of immunoreactive structures containing neurokinin in the human brainstem indicates that neurokinin might be involved in several physiological mechanisms, acting as a neurotransmitter and/or neuromodulator.

Coveńas Rafael; Martin Francisco; Belda Magdalena; Smith Victor; Salinas Pablo; Rivada Eva; Diaz-Cabiale Zaida; Narvaez Jose; Marcos Pilar; Tramu Gerard; Gonzalez-Baron Salvador

2003-01-01

148

Serum immunoreactive calcitonin concentration in hepatocellular carcinoma  

International Nuclear Information System (INIS)

[en] 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

149

DCLK1 immunoreactivity in colorectal neoplasia.  

UK PubMed Central (United Kingdom)

INTRODUCTION: 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.

Gagliardi G; Goswami M; Passera R; Bellows CF

2012-01-01

150

Bombesin-like immunoreactivity in the nervous system of hydra  

DEFF Research Database (Denmark)

With immunocytochemical methods, nerve cells have been detected in Hydra attenuata containing bombesin-like immunoreactivity. These nerve cells are located in ectoderm of all body regions of the animal and are especially abundant in basal disk and tentacles. Radioimmunoassay of extracts of hydra demonstrated at least 0.2 pmol/g wet weight of bombesin-like immunoreactivity. The immunoreactive material elutes from Sephadex G-50 in a similar position to synthetic bombesin. The data show that bombesin-like peptides are among the phylogenetically oldest neuropeptides found so far.

Grimmelikhuijzen, C J; Dockray, G J

1981-01-01

151

Evidence for white matter abnormalities in schizophrenia.  

UK PubMed Central (United Kingdom)

PURPOSE OF REVIEW: The purpose of this review is to highlight important recent imaging, histological, and genetic findings relevant to white matter abnormalities in schizophrenia. It is cast within the context of research findings conducted over the last 5 years, where we analyze their importance in understanding schizophrenia, as well as discuss future directions for research. RECENT FINDINGS: White matter abnormalities have long been hypothesized in schizophrenia, although only recently has it become possible to investigate them more closely. This has come about as a result of advances in neuroimaging, including new imaging techniques sensitive to white matter structure, as well as advances in computer science, with new analysis techniques making it possible to evaluate several interconnected brain regions at a time. Postmortem studies, with advances such as fluoroscopy and electron microscopy, have also led to quantifying populations of different brain cells, including myelin-forming oligodendrocytes. Moreover, molecular studies enable examination of immunoreactivity of proteins that are responsible for building myelin sheaths. Additionally, microarray genetic studies allow us to investigate myelin-related genes in schizophrenia. Taken together, these technological advances bring us closer to understanding white matter pathology in schizophrenia. SUMMARY: Advances in new imaging techniques likely account for the renewed interest in investigating white matter abnormalities in schizophrenia, with over 30 new articles published on this topic in the last 12 months, compared with 11 the year before. We review recent imaging, histological, and genetic findings that suggest white matter abnormalities in schizophrenia.

Kubicki M; McCarley RW; Shenton ME

2005-03-01

152

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

Gagliardi G; Goswami M; Passera R; Bellows CF

2012-01-01

153

Rapid renal clearance of immunoreactive canine plasma myoglobin  

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Rates and mechanisms of myoglobin removal from plasma were examined in closed-chest dogs, using disappearance patterns of immunoreactivity and radioactivity after i.v. canine myoglobin radiolabeled with 125I. Arterial immunoreactive myoglobin concentration decreased monoexponentially over a 2-decade range, with rate constants of disappearance averaging -0.080 +/- 0.014 min-1 (+/- SD). Renal arteriovenous difference in immunoreactive myoglobin concentration documented extraction of the parent molecule, with extraction ratios averaging 0.36 +/- 0.06. Renal venous specific activity increased a few minutes after myoglobin administration, consistent with discharge from the kidney of nonimmunoreactive radiolabeled peptides of the parent molecule. Arterial disappearance of 125I was subsequently delayed in relation to immunoreactive myoglobin. Urinary recoveries of immunoreactive parent molecule and radiolabeled constituents were limited, averaging 2.5 +/- 1.1% and 12 +/- 1.1% over a 6-hour period. Arterial rate, constants of disappearance of immunoreactive myoglobin decreased markedly with decreases in renal perfusion produced by obstruction of renal arterial inflow. Researchers conclude that myoglobin entering the vascular space is normally cleared rapidly by renal catabolism. Seven myoglobin concentration-time patterns during acute myocardial infarction directly reflect patterns of protein entry into the vascular space after release from injured tissue.

Klocke, F.J.; Copley, D.P.; Krawczyk, J.A.; Reichlin, M.

1982-06-01

154

Mechanism of action of cysteamine in depleting prolactin immunoreactivity  

International Nuclear Information System (INIS)

The thiol reagent cysteamine (CSH) depletes anterior pituitary cells of immunoreactive PRL both in vivo and in vitro. The authors examined the hypothesis that CSH affects either the solubility or immunoreactivity of PRL through a mechanism involving thiol-disulfide exchange. Adult female rats were treated with either CSH (300 mg/kg, sc) or an equimolar dose of ethanolamine as a control. Anterior pituitary glands were extracted in 0.1 M sodium borate buffer, pH 9.0. Treatment of pituitary extracts with beta-mercaptoethanol (BME) destroys the immunoreactivity of PRL. However, extraction in the presence of reduced glutathione or CSH of pituitaries of rats treated with CSH restores immunoreactive PRL to control levels. Extracts were also subjected to polyacrylamide gel electrophoresis (PAGE). On gels of pituitary extracts of CSH-treated rats, the band that comigrates with purified PRL is diminished compared to that in ethanolamine-treated controls. However, extraction of the pituitaries in sodium dodecyl sulfate-containing buffer followed by chemical reduction with BME restores the PRL band. Therefore, CSH acts on PRL through a thiol-related mechanism to yield a product that is poorly soluble in aqueous buffer at pH 9 and is poorly immunoreactive. Dispersed anterior pituitary cells in tissue culture were incubated with L-[35S]methionine to radiolabel newly synthesized peptides. PAGE followed by autoradiography confirmed the above results obtained in vivo

1985-01-01

155

Mechanism of action of cysteamine in depleting prolactin immunoreactivity  

Energy Technology Data Exchange (ETDEWEB)

The thiol reagent cysteamine (CSH) depletes anterior pituitary cells of immunoreactive PRL both in vivo and in vitro. The authors examined the hypothesis that CSH affects either the solubility or immunoreactivity of PRL through a mechanism involving thiol-disulfide exchange. Adult female rats were treated with either CSH (300 mg/kg, sc) or an equimolar dose of ethanolamine as a control. Anterior pituitary glands were extracted in 0.1 M sodium borate buffer, pH 9.0. Treatment of pituitary extracts with beta-mercaptoethanol (BME) destroys the immunoreactivity of PRL. However, extraction in the presence of reduced glutathione or CSH of pituitaries of rats treated with CSH restores immunoreactive PRL to control levels. Extracts were also subjected to polyacrylamide gel electrophoresis (PAGE). On gels of pituitary extracts of CSH-treated rats, the band that comigrates with purified PRL is diminished compared to that in ethanolamine-treated controls. However, extraction of the pituitaries in sodium dodecyl sulfate-containing buffer followed by chemical reduction with BME restores the PRL band. Therefore, CSH acts on PRL through a thiol-related mechanism to yield a product that is poorly soluble in aqueous buffer at pH 9 and is poorly immunoreactive. Dispersed anterior pituitary cells in tissue culture were incubated with L-(TVS)methionine to radiolabel newly synthesized peptides. PAGE followed by autoradiography confirmed the above results obtained in vivo.

Sagar, S.M.; Millard, W.J.; Martin, J.B.; Murchison, S.C.

1985-08-01

156

Immunoreactive glucagon responses to intravenous tolbutamide in chronic pancreatitis.  

UK PubMed Central (United Kingdom)

The effects of tolbutamide infusion (1 gm. over forty minutes) on plasma pancreatic glucagon-like immunoreactivity (PGLI), serum insulin, and blood glucose were studied in six patients with chronic pancreatitis and six matched controls.asal PGLI levels were significantly higher in the patients, despite higher fasting glucose concentrations. Tolbutamide infusion had no significant effect on mean PGLI levels in controls but was associated with significant elevation in pancreatitis patients, despite higher circulating glucose levels in the latter. The data suggest that chronic calcific pancreatitis patients hypersecrete immunoreactive glucagon, possibly from a nonpancreatic source and that this immunocreactive material may be stimulated by sulfonylureas.

Kalk WJ; Vinik AI; Paul M; Leller P; Jackson WP

1975-09-01

157

ABNORMAL PROTEIN REMOVING METHOD  

UK PubMed Central (United Kingdom)

A method for treating symptoms or diseases associated with accumulation of abnormal protein in the body includes administrating to a subject desiring such treatment a composition containing silybin and soybean saponin in an amount effective to reduce or remove abnormal protein produced in the body.

MIYATA SATOSHI; UMINO YUKARI

158

Structurally abnormal human autosomes  

Energy Technology Data Exchange (ETDEWEB)

Chapter 25, discusses structurally abnormal human autosomes. This discussion includes: structurally abnormal chromosomes, chromosomal polymorphisms, pericentric inversions, paracentric inversions, deletions or partial monosomies, cri du chat (cat cry) syndrome, ring chromosomes, insertions, duplication or pure partial trisomy and mosaicism. 71 refs., 8 figs.

NONE

1993-12-31

159

Abnormal rubbing and keratectasia.  

UK PubMed Central (United Kingdom)

PURPOSE: Hypotheses for the varied pathogenesis of the different forms of keratoconus have been outlined. Against this background, the possibility that abnormal rubbing causes or contributes to the development or progression of some forms of keratoconus has been examined. METHODS: Circumstantial evidence that shows an association between abnormal rubbing and keratoconus is reviewed, and a wide range of different forms of abnormal rubbing is described. Also examined is evidence of several processes whereby the cornea appears to be, or could be, adversely affected by mechanical trauma caused by rubbing. Conditions that may increase susceptibility to mechanical rubbing trauma have been discussed. RESULTS: Evidence of a role for inflammatory mediators in the pathogenesis of keratoconus appears to void the description of keratoconus as a noninflammatory condition. When vigorous knuckle-rubbing forces are located on the normal peripheral cornea, the thinner or weakened cone apex may be exposed to high intraocular pressure distending forces that may tend to promote ectasia. CONCLUSIONS: It appears reasonable to conclude that abnormal rubbing is a cause of some types of keratoconus, not because all abnormal rubbing, or only abnormal rubbing, leads to the development of some types of keratoconus, but because abnormal rubbing may increase the likelihood of the development of some forms of keratoconus. Abnormal rubbing habits may commence or continue after routine contact lens wear is established. Any associated rubbing or contact lens trauma may contribute to the progression of keratoconus. The abnormal rubbing-ectasia association in keratoconus may extend to other forms of keratectasia, including that seen after laser in situ keratomileusis, for which a contributory abnormal rubbing hypothesis may be appropriate.

McMonnies CW

2007-11-01

160

Neurotensin-like immunoreactivity after intestinal resection in the rat.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Neurotensin is a tridecapeptide located mainly in the distal small intestine. The present study was carried out in order to investigate the neurotensin response after proximal small intestinal resection in the rat. After resection, the median plasma concentration of neurotensin like immunoreactivity...

Olsen, P S; Pedersen, J H; Poulsen, S S; Yamashita, Y; Kirkegaard, P

 
 
 
 
161

Immunoreactivity of 125I-papain labelled by different methods  

International Nuclear Information System (INIS)

[en] Three different methods of papain iodination (with chloramine-T, lactoperoxidase and conjugation with Bolton-Hunter reagent) have been compared. The highest yield of 125I-papain could be obtained using lactoperoxidase which enabled to achieve the highest immunoreactivity. 125I-papain, labelled this way, is suitable for the radioimmunoassay of papain. (author)

1984-01-31

162

Cloning and Expression of Immunoreactive Antigens from Mycobacterium tuberculosis  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Four immunoreactive proteins, B.4, B.6, B.10, and B.M, with molecular weights ranging from 16,000 to 58,000, were observed from immunoblots of Mycobacterium tuberculosis total lysates screened with sera from individuals with active tuberculosis. These proteins were identified from microsequence anal...

Hong Lim, Renee Lay; Kiang Tan, Li; Fun Lau, Wai; Ching Ming , Maxey; Chung; Dunn, Roseanne; Phon Too, Heng; Chan, Lily

163

Studies in abnormal pressures  

CERN Document Server

When Fertl's first book, Abnormal Formation Pressures, was published by Elsevier in 1976, the topic was relatively new in book form. In the years that followed, his book became the standard work for petroleum engineers and drillers. The list of major petroleum provinces with abnormally high pore pressures has grown steadily over the years, and with it has grown our knowledge and experience. There have also been technological advances. A new book was required, but no longer could the topic be covered adequately by one person. The problems of abnormally high formation pressures encount

Fertl, WH; Hotz, RF

1994-01-01

164

Plasma gastrin immunoreactivity in dogs with acute gastric dilatation-volvulus.  

UK PubMed Central (United Kingdom)

Plasma gastrin immunoreactivity was measured by radioimmunoassay in 45 dogs with acute gastric dilatation-volvulus (GDV). Significant increases (P less than 0.05) were found in dogs with acute GDV and in the fasted state after surgical treatment and recovery. The data suggested that dogs that have had GDV may have preexisting high plasma gastrin immunoreactivity. In dogs with acute GDV, plasma gastrin immunoreactivity was not found to be helpful in formulating prognosis. Circumcostal gastropexy did not affect plasma gastrin immunoreactivity.

Leib MS; Wingfield WE; Twedt DC; Bottoms GD

1984-07-01

165

[Immunoreactive parathyroid hormone in samples from thyroid veins (author's transl)  

UK PubMed Central (United Kingdom)

Blood samples were obtained from 16 patients with primary or secondary hyperparathyroidism by selectively catheterising cervical and thyroid veins. Immunoreactive parathyroid hormone was measured with two aminoterminal (anti-N)- and three carboxyl-terminal (anti-C)-specific antisera. Two anti-C sera and one anti-N serum were useful in localising the parathyroid tumors. In addition, the parathyroid hormone fragments were separated by gelfiltration and the immunoreactivity in the effluent was estimated with anti-N and anti-C sera. The results suggest that the usefulness of antisera for parathyroid tissue localisation is determined by their affinity for the intact hormone, regardless of their anti-N or anti-C qualities.

Offermann G; Sörensen R; Häring R

1978-10-01

166

Increased caspase-3 immunoreactivity of erythrocytes in STZ diabetic rats.  

UK PubMed Central (United Kingdom)

Eryptosis is a term to define apoptosis of erythrocytes. Oxidative stress and hyperglycemia, both of which exist in the diabetic intravascular environment, can trigger eryptosis of erythrocytes. In this experimental study, it is presented that the majority of erythrocytes shows caspase-3 immunoreactivity in streptozocin- (STZ)-induced diabetic rats. Besides that, caspase-3 positive erythrocytes are aggregated and attached to vascular endothelium. In conclusion, these results may start a debate that eryptosis could have a role in the diabetic complications.

F?rat U; Kaya S; Cim A; Büyükbayram H; Gökalp O; Dal MS; Tamer MN

2012-01-01

167

Inhibin-A immunoreactivity in nervous system lesions.  

UK PubMed Central (United Kingdom)

To evaluate inhibin-A immunoreactivity and its utility in the differential diagnosis of nervous system neoplasms and non-neoplastic lesions. An immunohistochemical study of 252 central and peripheral nervous system tumors and 40 non-neoplastic lesions was undertaken. Brain lesions included the basic spectrum of astrocytic, oligodendroglial, and ependymal neoplasms, as well as glioneuronal, pineal parenchymal, choroid plexus, and embryonal. Meningeal neoplasms, basic peripheral nerve tumors, and uncommon sellar lesions were also assessed. Non-neoplastic lesions included demyelinating disease, progressive multifocal leukoencephalopathy, organizing infarct, and reactive gliosis. Diffuse cytoplasmic, membranous, and perinuclear cytoplasmic staining patterns were observed. Significant immunoreactivity was noted in glioblastoma (12 of 20), pleomorphic xanthoastrocytoma (6 of 10), ganglioglioma (8 of 10), meningioma (14 of 20), and hemangioblastoma (10 of 10). Peripheral nerve and sellar tumors as well as non-neoplastic lesions were entirely immunonegative. In our study that investigated the inhibin-A immunoreactivity in a broad spectrum of nervous system lesions, inhibin-A positivity was established in various low-grade and high-grade central nervous system tumors. Thus, inhibin-A is not a specific marker of hemangioblastoma and may be of limited utility in the differential diagnosis of astrocytic and meningothelial neoplasms. Its pathophysiologic role in these various tumors remains to be determined. Further evaluation of the possible significance of staining patterns and degrees of reactivity relative to pathobiology and/or prognosis significance is required.

Gurses I; Scheithauer BW

2012-05-01

168

Inhibin-A immunoreactivity in nervous system lesions.  

Science.gov (United States)

To evaluate inhibin-A immunoreactivity and its utility in the differential diagnosis of nervous system neoplasms and non-neoplastic lesions. An immunohistochemical study of 252 central and peripheral nervous system tumors and 40 non-neoplastic lesions was undertaken. Brain lesions included the basic spectrum of astrocytic, oligodendroglial, and ependymal neoplasms, as well as glioneuronal, pineal parenchymal, choroid plexus, and embryonal. Meningeal neoplasms, basic peripheral nerve tumors, and uncommon sellar lesions were also assessed. Non-neoplastic lesions included demyelinating disease, progressive multifocal leukoencephalopathy, organizing infarct, and reactive gliosis. Diffuse cytoplasmic, membranous, and perinuclear cytoplasmic staining patterns were observed. Significant immunoreactivity was noted in glioblastoma (12 of 20), pleomorphic xanthoastrocytoma (6 of 10), ganglioglioma (8 of 10), meningioma (14 of 20), and hemangioblastoma (10 of 10). Peripheral nerve and sellar tumors as well as non-neoplastic lesions were entirely immunonegative. In our study that investigated the inhibin-A immunoreactivity in a broad spectrum of nervous system lesions, inhibin-A positivity was established in various low-grade and high-grade central nervous system tumors. Thus, inhibin-A is not a specific marker of hemangioblastoma and may be of limited utility in the differential diagnosis of astrocytic and meningothelial neoplasms. Its pathophysiologic role in these various tumors remains to be determined. Further evaluation of the possible significance of staining patterns and degrees of reactivity relative to pathobiology and/or prognosis significance is required. PMID:22505009

Gurses, Iclal; Scheithauer, Bernd W

2012-05-01

169

Abnormal Gait Recognition  

Directory of Open Access Journals (Sweden)

Full Text Available Due to increasing crime rate identification using biometrics has become an important field of research. When it is not possible to take snapshot, to read iris, to take finger prints etc then identification using gait may be proved an effective tool to identify a person. This paper presents a method which distinguishs between normal and abnormal gait. A person having abnormal gait may be categorize as suspicious and alarming actions may be taken. Experiments have been done on real world data and system has been trained for normal walk for real world subjects.

Naveen Rohila; Prof. Brijesh Kumar,; Naresh Chauhan

2010-01-01

170

Models of Abnormal Scarring.  

UK PubMed Central (United Kingdom)

Keloids and hypertrophic scars are thick, raised dermal scars, caused by derailing of the normal scarring process. Extensive research on such abnormal scarring has been done; however, these being refractory disorders specific to humans, it has been difficult to establish a universal animal model. A wide variety of animal models have been used. These include the athymic mouse, rats, rabbits, and pigs. Although these models have provided valuable insight into abnormal scarring, there is currently still no ideal model. This paper reviews the models that have been developed.

Seo BF; Lee JY; Jung SN

2013-01-01

171

Consanguinity and chromosomal abnormality  

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Full Text Available BACKGROUND : Consanguinity is defined as the marriage between close relatives. The deleterious effects associated with consanguinity may be caused by the expression of rare recessive genes inherited from common ancestors. AIMS AND OBJECTIVES : The present study was undertaken to analyze the effect of consanguinity on chromosomal abnormality (CA). METHODS AND MATERIALS : During last 6 years period, a total of 1465 cases with suspected genetic etiology like bad obstetric history, mental retardation, multiple congenital anomalies, Down syndrome, primary amenorrhea and primary infertility was referred to Division of Human Genetics for karyotyping and genetic counseling. The information regarding consanguinity was obtained through pedigree analyzes up to three generations from all the patients. Chi-square test was applied to test the significance. RESULTS : Consanguinity was seen in 427 cases (29.14%), 305 cases were confirmed to have CA, among them 240 (78.7%) had numerical abnormality and 65 (21.3%) had structural abnormality. The presence of consanguinity in CA was seen in 53 cases (17%), including 43 (81.1%) with numerical and 10 (18.9%) with structural abnormality. CONCLUSION : The effect of consanguinity on CA was almost significant ( P < 0.001), whereas the effect was not significant for the type of CA. It may be because of the pooled types of consanguinity as well as the CA. Further information is needed to state categorically that there could be the effect of consanguinity on CA.

Amudha S; Aruna N; Rajangam S

2005-01-01

172

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

173

Abnormality, rationality, and sanity.  

UK PubMed Central (United Kingdom)

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.

Hertwig R; Volz KG

2013-09-01

174

Gamma-aminobutyric acid B receptor immunoreactivity in the mouse adrenal medulla.  

UK PubMed Central (United Kingdom)

The present study examined gamma-aminobutyric acid B (GABAB ) receptor, GABA, choline acetyltransferase (ChAT), and neuronal nitric oxide synthase (nNOS) immunoreactivities in the mouse adrenal medulla. GABAB receptor immunoreactivity was seen in numerous chromaffin cells and in a few ganglion cells of the adrenal medulla. By using a formaldehyde-induced fluorescence (FIF) method, GABAB receptor immunoreactivity was observed in numerous adrenaline (A) cells, but not in noradrenaline (NA) cells showing blue-white fluorescence. This suggests that GABAB receptors may be present in the A cells and be related to the secretory activity of A cells but not NA cells in the mouse adrenal medulla. GABAB receptor immunoreactive ganglion cells were shown to be nNOS immunopositive by using a double immunostaining method. Weak GABA immunoreactivity was visible in some chromaffin cells and in the numerous nerve fibers of the medulla. By using the FIF method, weak GABA-immunoreactive chromaffin cells were shown to be in the NA cells showing blue-white fluorescence. GABA-immunoreactive nerve fibers were in dense contact in A cells, but not NA cells. GABA-immunoreactive nerve fibers closely contacted a few ganglion cells. Numerous GABA-immunoreactive nerve fibers in the medulla showed ChAT immunoreactive. This result suggests that GABA and acetylcholine may be released from the same nerve fibers and may have a secretory effect on the A cells of the medulla.

Oomori Y; Murabayashi H; Kuramoto H; Kawano H; Kato K; Nakagawa C; Sasaki M; Kitamura N; Ishikawa K; Tanaka K

2013-06-01

175

Pathologic and clinical features of pituitary adenomas showing TSH immunoreactivity.  

UK PubMed Central (United Kingdom)

Patients with thyrotropin-secreting pituitary adenomas may present with mass effect, hypopituitarism, and/or hyperthyroidism. The spectrum of pathologic and clinical features of patients whose tumors demonstrate ?-thyrotropin immunoreactivity (?-TSH IR) has not been characterized. To characterize the phenotype of patients with pituitary adenomas with positive ?-TSH IR, we conducted a retrospective analysis of patient records of all adult patients (n = 1,223) undergoing pituitary surgery in our institution over one decade (1999-2009). The search identified 166 adults with tumors which had ?-TSH IR. These patients were individually matched to 166 patients whose tumors revealed no ?-TSH IR. Clinical, pathological, imaging and biochemical data were extracted. 332 patients, aged 51.4 ± 15.1 years [150 women (45 %) and 182 men (55 %)], with pituitary adenomas (mean tumor diameter ± SD: 22.7 ± 9.0 mm) were studied. The degree of ?-TSH IR was associated with the presence of central hyperthyroidism (p < 0.0001) or goiter (p = 0.0217). Patients whose tumors expressed more extensive ?-TSH IR were less likely to develop pituitary apoplexy than those without ?-TSH IR (p = 0.0428). In addition, the degree of ?-TSH IR correlated with the presence of immunoreactivity for ?-FSH (p < 0.0001), ?-LH (p < 0.0001), alpha subunit (p < 0.0001), and GH (p = 0.0036). Conclusions: Pituitary adenomas expressing ?-TSH IR were more likely to demonstrate immunoreactivity for ?-FSH, ?-LH, GH or alpha subunit. Patients with such tumors were more likely to exhibit hyperthyroidism and goiter, but less likely to develop pituitary apoplexy than patients without ?-TSH IR. These findings suggest that ?-TSH IR is associated with specific phenotypic features in patients with pituitary adenomas.

Tritos NA; Eppakayala S; Swearingen B; Hedley-Whyte ET; Miller KK; Nachtigall LB; Grinspoon SK; Biller BM; Klibanski A

2013-09-01

176

Enzyme immunoassay of immunoreactive trypsin in serum and blood spots  

International Nuclear Information System (INIS)

[en] An enzyme immunoassay method for the assay of serum immunoreactive trypsin (IRT) is described. The method is a two site binding assay carried out on microtitre plates as the solid phase. Wells were coated with affinity purified anti-human trypsin and bioinylated anti-trypsin and avidin-?-galactosidase were used as the second antibody and detection system respectively. The assay was sensitive enough to determine IRT concentrations in either serum or dried blood spots. A good correlation was obtained when the method was compared with the Hoechst radioimmunoassay method. (Author)

1988-01-01

177

Immunoreactivity examination of patients with testicular tumours treated with radiotherapy  

International Nuclear Information System (INIS)

[en] 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)

1985-01-01

178

Liver abnormalities in pregnancy.  

UK PubMed Central (United Kingdom)

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.

Than NN; Neuberger J

2013-08-01

179

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

180

Small skeletal abnormalities  

International Nuclear Information System (INIS)

[en] To date, the skeletal imaging capabilities of digital radiography with storage phosphors have been poorly investigated, and the diagnostic accuracy ofthis technique has not been thoroughly assessed. To evaluate the performance of storage phosphor digital radiography we compared 66 conventional and 66 digital radiographs of small abnormalities of the extremities (fractures, erosions, calcifications). Conventional images were obtained with a low-speed screen-film system while digital ones were acquired with high resolution (5lp/mm max) phosphors and laser-printed on a 8'x10' film. Two experienced radiologist defined the gold standard (389 abnormalities) and four radiologist scored the findings (1,556 observations) on a five-point discrete scale. ROC analysis indicated film and storage radiography to be equally effective in the overall detection of abnormalities. No difference was found in the individual performances of the four readers in the site subclasses (wrist, hand) and in the specific detection of fractures and erosions. Digital radiography proved to be superior to conventional radiography in the detection of calcifications in all sites and particularly in the wrist (p

1991-01-01

 
 
 
 
181

Presence and distribution of serotonin immunoreactivity in the cyprids of the barnacle Balanus amphitrite  

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Full Text Available In this work, the presence and distribution of serotonin in the cyprid of the barnacle Balanus amphitrite were investigated by immunohistochemical methods. Serotonin-like immunoreactive neuronal cell bodies were detected in the central nervous system only. Various clusters of immunoreactive neuronal cell bodies are distributed in the brain (protocerebrum, deutocerebrum, optical lobes), and at least, four pairs of neuronal cell bodies were detected in the centrally positioned neuropil of the posterior ganglion. Rich plexuses of immunoreactive nerve fibers in the neuropil area were also observed. Furthermore, bundles of strongly immunoreactive nerve fibers surrounding the gut wall were localized, and immunoreactive nerve terminals in the antennules and compound eyes were observed. These data demonstrate the presence of a serotonin-like immunoreactive substance in the barnacle cyprids; furthermore, its immunolocalization in the cephalic nerve terminals allows us to postulate the involvement of this bioactive molecule in substrate recognition during the settlement process.

L Gallus; P Ramoino; M Faimali; V Piazza; G Maura; M Marcoli

2005-01-01

182

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

Directory of Open Access Journals (Sweden)

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; Özlem CANÖZ; Özlem ER; Kemal DEN?Z; Serdar SOYUER

2002-01-01

183

Immunoreactivity of anti-gelsolin antibodies: implications for biomarker validation  

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Full Text Available Abstract Background Proteomic-based discovery of biomarkers for disease has recently come under scrutiny for a variety of issues; one prominent issue is the lack of orthogonal validation for biomarkers following discovery. Validation by ELISA or Western blot requires the use of antibodies, which for many potential biomarkers are under-characterized and may lead to misleading or inconclusive results. Gelsolin is one such biomarker candidate in HIV-associated neurocognitive disorders. Methods Samples from human (plasma and CSF), monkey (plasma), monocyte-derived macrophage (supernatants), and commercial gelsolin (recombinant and purified) were quantitated using Western blot assay and a variety of anti-gelsolin antibodies. Plasma and CSF was used for immunoaffinity purification of gelsolin which was identified in eight bands by tandem mass spectrometry. Results Immunoreactivity of gelsolin within samples and between antibodies varied greatly. In several instances, multiple bands were identified (corresponding to different gelsolin forms) by one antibody, but not identified by another. Moreover, in some instances immunoreactivity depended on the source of gelsolin, e.g. plasma or CSF. Additionally, some smaller forms of gelsolin were identified by mass spectrometry but not by any antibody. Recombinant gelsolin was used as reference sample. Conclusions Orthogonal validation using specific monoclonal or polyclonal antibodies may reject biomarker candidates from further studies based on misleading or even false quantitation of those proteins, which circulate in various forms in body fluids.

Haverland Nicole; Pottiez Gwënaël; Wiederin Jayme; Ciborowski Pawel

2010-01-01

184

Neurodevelopmental abnormalities in ADHD.  

UK PubMed Central (United Kingdom)

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 factors, such as symptom remission/persistence and exposure to stimulant treatment. Findings from functional imaging studies point to multiple loci of abnormalities that are not limited to frontal-striatal circuitry, which is important for executive and motivational function, but also include parietal, temporal and motor cortices, and the cerebellum. However, a definitive conclusion about maturational delays or alternate trajectories cannot be drawn from this work as activation patterns are influenced by task-specific factors that may induce variable performance levels and strategies across development. In addition, no studies have implemented cross-sectional or longitudinal designs, without which the developmental origin of differences in activation cannot be inferred. Thus, current task-evoked functional imaging provides information about dynamic or state-dependent differences rather than fixed or trait-related differences. In the future, task-free functional imaging holds promise for revealing neurodevelopmental information that is minimally influenced by performance/strategic differences. Further, studies using longitudinal designs that identify sources of phenotypic heterogeneity in brain maturation and characterize the relationship between brain function and underlying structural properties are needed to provide a comprehensive view of neurodevelopmental abnormalities in ADHD.

Vaidya CJ

2012-01-01

185

Russia: An Abnormal Country  

Directory of Open Access Journals (Sweden)

Full Text Available Andrei Shleifer and Daniel Treisman recently rendered a summary verdict on the post Soviet Russian transition experience finding that the Federation had become a normal country with the west's assistance, and predicting that it would liberalize and develop further like other successful nations of its type. This essay demonstrates that they are mistaken on the first count, and are likely to be wrong on the second too. It shows factually, and on the norms elaborated by Pareto, Arrow and Bergson that Russia is an abnormal political economy unlikely to democratize, westernize or embrace free enterprise any time soon

Steven Rosefielde

2005-01-01

186

Minicolumnar abnormalities in autism.  

UK PubMed Central (United Kingdom)

Autism is characterized by qualitative abnormalities in behavior and higher order cognitive functions. Minicolumnar irregularities observed in autism provide a neurologically sound localization to observed clinical and anatomical abnormalities. This study corroborates the initial reports of a minicolumnopathy in autism within an independent sample. The patient population consisted of six age-matched pairs of patients (DSM-IV-TR and ADI-R diagnosed) and controls. Digital micrographs were taken from cortical areas S1, 4, 9, and 17. The image analysis produced estimates of minicolumnar width (CW), mean interneuronal distance, variability in CW (V (CW)), cross section of Nissl-stained somata, boundary length of stained somata per unit area, and the planar convexity. On average CW was 27.2 microm in controls and 25.7 microm in autistic patients (P = 0.0234). Mean neuron and nucleolar cross sections were found to be smaller in autistic cases compared to controls, while neuron density in autism exceeded the comparison group by 23%. Analysis of inter- and intracluster distances of a Delaunay triangulation suggests that the increased cell density is the result of a greater number of minicolumns, otherwise the number of cells per minicolumns appears normal. A reduction in both somatic and nucleolar cross sections could reflect a bias towards shorter connecting fibers, which favors local computation at the expense of inter-areal and callosal connectivity.

Casanova MF; van Kooten IA; Switala AE; van Engeland H; Heinsen H; Steinbusch HW; Hof PR; Trippe J; Stone J; Schmitz C

2006-09-01

187

Minicolumnar abnormalities in autism.  

Science.gov (United States)

Autism is characterized by qualitative abnormalities in behavior and higher order cognitive functions. Minicolumnar irregularities observed in autism provide a neurologically sound localization to observed clinical and anatomical abnormalities. This study corroborates the initial reports of a minicolumnopathy in autism within an independent sample. The patient population consisted of six age-matched pairs of patients (DSM-IV-TR and ADI-R diagnosed) and controls. Digital micrographs were taken from cortical areas S1, 4, 9, and 17. The image analysis produced estimates of minicolumnar width (CW), mean interneuronal distance, variability in CW (V (CW)), cross section of Nissl-stained somata, boundary length of stained somata per unit area, and the planar convexity. On average CW was 27.2 microm in controls and 25.7 microm in autistic patients (P = 0.0234). Mean neuron and nucleolar cross sections were found to be smaller in autistic cases compared to controls, while neuron density in autism exceeded the comparison group by 23%. Analysis of inter- and intracluster distances of a Delaunay triangulation suggests that the increased cell density is the result of a greater number of minicolumns, otherwise the number of cells per minicolumns appears normal. A reduction in both somatic and nucleolar cross sections could reflect a bias towards shorter connecting fibers, which favors local computation at the expense of inter-areal and callosal connectivity. PMID:16819561

Casanova, Manuel F; van Kooten, Imke A J; Switala, Andrew E; van Engeland, Herman; Heinsen, Helmut; Steinbusch, Harry W M; Hof, Patrick R; Trippe, Juan; Stone, Janet; Schmitz, Christoph

2006-07-04

188

[Penile congenital abnormalities].  

UK PubMed Central (United Kingdom)

INTRODUCTION: Congenital abnormalities of the penis are usually diagnosed at birth and pose aesthetic and functional problems sometimes requiring surgical management. METHODS: A literature review was conducted on Medline considering the articles listed until January 2012. RESULTS: Hypospadias is the most common malformation (1 in 250 boys. Familial forms: 7%). The causes remain hypothetical but the doubling of the incidence in 30 years could be linked to fetal exposure to endocrine disruptors "estrogen-like" used in the food industry in particular. Surgical treatment is usually intended to improve the aesthetic appearance but sometimes, in case of significant curvature or posterior meatus, necessary for normal sexual life and fertility. Other malformations (epispades, buried penis, transpositions, twists and preputial abnormalities) as well as management for functional or aesthetic consequences of these malformations in adulthood require complex surgical care in a specialized environment. CONCLUSION: The improvement of surgical techniques and pediatric anesthesia allows an early and effective specialized surgical approach of penile malformations. Management of sequelae in adulthood must be discussed and requires experience of surgical techniques on pediatric and adult penis.

Boillot B; Teklali Y; Moog R; Droupy S

2013-07-01

189

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

190

[Immunoreaction and blood transfusion--chairmen's introductory remarks].  

UK PubMed Central (United Kingdom)

Although blood transfusion is an extremely important therapeutic procedure that usually proceeds without complications, there are some risks associated with donated blood. Investigations into the causes of transfusion reactions and their prevention are important issues for transfusion therapy. In addition to nucleic acid amplification testing (NAT) for infectious diseases and the irradiation of blood to prevent post-transfusion GVHD, prestorage leukocyte reduction and diversion of the first part of the donation of blood were recently introduced into transfusion therapy. This symposium, entitled "Immunoreaction and blood transfusion", reviewed the immune responses associated with blood transfusion, which is probably the most frequent medical procedure performed in allogeneic organ transplantation, with four themes provided by the four featured invited speakers: transfusion-related acute lung injury (TRALI) and transfusion-associated circulatory overload (TACO), high-dose intravenous immunoglobulin therapy for chronic inflammatory demyelinating polyradiculoneuropathy, transfusion-transmitted infectious disease surveillance, and transfusion-related immunomodulation.

Kawabe T; Matsushita T

2013-05-01

191

Exocrine secretion of immunoreactive erythropoietin from the rat submaxillary gland.  

UK PubMed Central (United Kingdom)

The physiological role of immunoreactive erythropoietin (iEp) in rodent submaxillary glands (SMG) is largely unknown. We studied in vivo the effects of cholinergic and adrenergic agents in male rats with respect to exocrine secretion of iEp into saliva. Intravenous administration of metacholine (20 micrograms/kg), norepinephrine (30 micrograms/kg), and isoproterenol (30 micrograms/kg) resulted in equal volumes of saliva over 1 h. None of the drugs altered circulating plasma levels and kidney concentrations of iEp. Salivary secretions induced by either norepinephrine or isoproterenol, both adrenergic agonists, contained high levels of iEp and a significant depletion of gland content was observed, suggesting that SMG exocrine iEp secretion is mediated by adrenergic receptors. In contrast, metacholine-stimulated glands retained their full iEp content and iEp was undetectable in saliva, indicating that cholinergic activity is not associated with exocrine secretion of iEp from SMGs.

Clemons GK; Elverdin JC; Allippi RM; Barcelo AC; Stefano FJ; Bozzini CE

1988-02-01

192

Parvalbumin-immunoreactive neurons in the human claustrum.  

UK PubMed Central (United Kingdom)

The morphology and distribution of parvalbumin-immunoreactive neurons (PV-ir) were studied in the human claustrum. PV-ir neurons were observed throughout the claustrum, with the highest numbers noted in the central (broadest) portion as compared with the dorsal and ventral aspects. Reaction product was evident in the neuronal perikarya, dendritic processes, and spines. In the majority of these labeled neurons, the cytoplasm was devoid of lipofuscin pigment. Cell bodies varied widely in both shape and size, ranging from oval and small, to multipolar and large. PV-ir neurons were classified into two groups, primarily based on dendritic morphology: spiny neurons with long and straight dendrites, and aspiny neurons with thin and curving dendritic processes. PV-ir fibers were seen throughout the neuropil, with many immuno-positive puncta noted.

Hinova-Palova DV; Edelstein L; Landzhov BV; Braak E; Malinova LG; Minkov M; Paloff A; Ovtscharoff W

2013-07-01

193

Digoxin-like immunoreactivity in human body fluids  

International Nuclear Information System (INIS)

[en] The clinical and chemical characteristics of a solid-phase radioimmunoassay (RIA) for routine digoxin determination has been studied with the aim to confirm our previous observation of the presence of digoxin-like immunoreactive substance (DLIS) in serum (plasma) and urine of normal subjects not under digoxin treatment. The sensitivity of the assay was 2.1±0.6 pg/tube and the reproducibility, tested with two different urine pools in terms of digoxin-equivalents (d.e.), was 12.5% (285.6±35.7 pg/ml d.e., n=19) and 20.6% (123.8±25.5 pg/ml d.e., n=19), respectively. The mean DLIS concentration in the blood of 32 normal subjects was 15.6±8.0 pg/ml d.e. (range 0-60 pg/ml d.e.). The mean DLIS concentration in urine of 37 normal subjects (overnight collection) was 160.0±52.3 pg/ml d.e. (range 70-350 pg/ml d.e.), while the mean 24-hour DLIS excretion of 10 normal subjects was 97.3±39.7 ng d.e. Two urine pools were extracted with organic solvents. Good recoveries (80-100%) were obtained with methanol, while poor recoveries were obtained with methylene chloride, hexane and petroleum ether. The present study indicates that DLIS is not a large charged molecule, neither salt, nor fatty acid, which are considered the most frequent non-specific interferences in RIA systems. Urine samples may be more useful for pathophysiological studies on digoxin-like immunoreactivity in human body fluids, because of their higher DLIS concentrations (4-10 times the concetration in blood)

1985-01-01

194

Parvalbumin immunoreactivity during the development of the cerebellum of the rainbow trout  

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The distribution of parvalbumin immunoreactivity in the developing cerebellum ofthe rainbow trout was studied by using a specific monoclonal antibody and the avidin-biotin peroxidase method. Parvalbumin immunoreactivity was absent during the embryonic development of the cerebellum. The first immunor...

Porteros Herrero, Ángel; Arévalo, Rosario; Brińón, Jesús G.; Crespo, Carlos; Aijón Noguera, José; Alonso Peńa, José-Ramón

195

Calcitonin gene-related peptide immunoreactive nerve fibers in the rat conjunctiva.  

Science.gov (United States)

Calcitonin gene-related peptide (CGRP) immunoreactive nerve fibers were studied in the rat conjunctiva by using indirect immunohistochemistry. Their origin was evaluated in a series of experiments where the animals were denervated by electrocoagulating the two first branches of the trigeminal nerve or by surgically extirpating the superior cervical ganglion. The CGRP-immunoreactive nerve fibers were seen mainly as thin varicose fibers in the epithelium and in the stroma. Many of the stromal fibers showed no apparent destination. However, CGRP-immunoreactive fibers were commonly found in association with stromal blood vessels, the smooth muscle of Müller, and the meibomian glands. Approximately 40% of the ganglion cells in the trigeminal ganglion were immunoreactive to CGRP. In the superior cervical ganglion, a few CGRP-immunoreactive fibers were seen although the ganglion cells were negative. After trigeminal denervation, all the epithelial and most of the stromal CGRP-immunoreactive nerve fibers disappeared. Sympathectomy had no effect on the presence of the CGRP-immunoreactive fibers. These observations indicate that most of the CGRP-immunoreactive nerve fibers in the rat conjunctiva are sensory nerves originating in the trigeminal ganglion. A few of the demonstrated fibers are, however, resistant to the sensory denervation and may be parasympathetic in their origin. PMID:2001937

Luhtala, J; Palkama, A; Uusitalo, H

1991-03-01

196

Calcitonin gene-related peptide immunoreactive nerve fibers in the rat conjunctiva.  

UK PubMed Central (United Kingdom)

Calcitonin gene-related peptide (CGRP) immunoreactive nerve fibers were studied in the rat conjunctiva by using indirect immunohistochemistry. Their origin was evaluated in a series of experiments where the animals were denervated by electrocoagulating the two first branches of the trigeminal nerve or by surgically extirpating the superior cervical ganglion. The CGRP-immunoreactive nerve fibers were seen mainly as thin varicose fibers in the epithelium and in the stroma. Many of the stromal fibers showed no apparent destination. However, CGRP-immunoreactive fibers were commonly found in association with stromal blood vessels, the smooth muscle of Müller, and the meibomian glands. Approximately 40% of the ganglion cells in the trigeminal ganglion were immunoreactive to CGRP. In the superior cervical ganglion, a few CGRP-immunoreactive fibers were seen although the ganglion cells were negative. After trigeminal denervation, all the epithelial and most of the stromal CGRP-immunoreactive nerve fibers disappeared. Sympathectomy had no effect on the presence of the CGRP-immunoreactive fibers. These observations indicate that most of the CGRP-immunoreactive nerve fibers in the rat conjunctiva are sensory nerves originating in the trigeminal ganglion. A few of the demonstrated fibers are, however, resistant to the sensory denervation and may be parasympathetic in their origin.

Luhtala J; Palkama A; Uusitalo H

1991-03-01

197

[Algodystrophy and metabolic abnormalities  

UK PubMed Central (United Kingdom)

The French Society of Rheumatology national study of reflex dystrophy revealed a serum glucose greater than 1.20 g/l in 9.09 p. cent of cases, a serum uric acid greater than 70 mg/l in 25 p. cent of men and greater than 60 mg/l in 14 p. cent of women. Serum cholesterol was normal in males, but higher than the mean + 2 sigma in 23 p. cent of females. Serum triglycerides were higher than m + 2 sigma in 55 p. cent of men and 56 p. cent of women. From a group of 80 patients, 54 (67.5 p. cent) had at least one of the three metabolic abnormalities and 49 (61.25 p. cent) were hypertriglyceridaemic. This hypertriglyceridaemia is the most frequent abnormality found. When hyperglycaemia or hyperuricaemia are present it is almost always in association with hypertriglyceridaemia. Hypertriglyceridaemia is more common in algoneurodystrophy of the lower limbs (54/78, 69 p. cent) than in algoneurodystrophy of the upper limbs (5/22, 22.7 p. cent). Algoneurodystrophy is more often primary, when it occurs in the lower limbs and post-traumatic when it occurs in the upper limbs. A prospective study comparing several parameters of lipid metabolism in 24 patients with algoneurodystrophy and 15 matched controls showed a significant drop in the HDL Chol/HDL P1 ratio and in the DHL - HDL Chol/TG ratio in patients with algoneurodystrophy. Serum insulin was comparable in the 2 groups. Lecithin cholesterol acyl transferase (LCAT), SGOT, SGPT and GGT were normal. The essential role of hypertriglyceridaemia in the genesis of the characteristic bony lesions of algoneurodystrophy is discussed.

Amor B; Tallet F; Raichvarg D; Guenee B; de Gery A; Damak A; Kharrat A; Ekindjian OG

1983-06-01

198

The distribution of sensory fibers immunoreactive for the TRPV1 (capsaicin) receptor in the human prostate.  

UK PubMed Central (United Kingdom)

OBJECTIVES: To determine the distribution of sensory fibers immunoreactive to the pain receptor TRPV1 in the human prostate. METHODS: Eight prostates were harvested from cadaver transplant donors and immediately immersion fixed. Longitudinal and transverse 20 microm sections were cut on a cryostat and immunoreacted with two anti-human TPRV1 antibodies. RESULTS: TRPV1-immunoreactive nerve fibers were distributed throughout the prostatic urethral mucosa, verumontanum, ejaculatory ducts and periurethral prostatic acini. In the urethral mucosa, TRPV1-immunoreactive fibers penetrated the epithelial layer up to its surface. In the transitional and peripheral zones of the gland no TRPV1-immunoreactive nerve fibers were detected. CONCLUSIONS: The existence of a rich TRPV1 sensory innervation in the human prostate may open new therapeutic perspectives for the treatment of pain in patients with chronic prostatitis (Chronic Pelvic Pain Syndrome).

Dinis P; Charrua A; Avelino A; Nagy I; Quintas J; Ribau U; Cruz F

2005-07-01

199

Heterogeneity of motilin-immunoreactive cells in the duodenum and pyloric region of several avian species.  

UK PubMed Central (United Kingdom)

Immunohistochemical characterizations of motilin-immunoreactive cells were examined in gastric and duodenal mucosae of nine species of birds from seven orders using five different region-specific motilin antisera. Motilin-immunoreactive cells appeared as open-type cells in the mucosal epithelium and showed varying immunoreactivities to antisera used in all the birds examined except for the cormorant and penguin, which did not show any kinds of immunoreactivity to motilin. Motilin-immunoreactive cells of the emu duodenum were detected by all the motilin-antisera used. The present results suggest that there is a wide range of heterogeneity between motilin molecules among avian species, or perhaps alternatively the existence of a family of motilin-like peptide. Furthermore, the present results should prove useful for a molecular biological study on the evolution of avian motilin.

Yamada J; Arita M; Kitamura N; Yamashita T; Yanaihara N; Richardson KC

1993-08-01

200

Protein aggregation and defective RNA metabolism as mechanisms for motor neuron damage.  

Science.gov (United States)

The presence of protein inclusions within the central nervous system is a characteristic of most neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS). Aggregates may induce cell death trough several mechanisms, such as sequestration of essential cellular components, clogging of the proteasome system, and/or disruption of axonal transport. The neuropathological signature of ALS is represented by the presence of ubiquitinated inclusions immunoreactive for the protein TDP-43 in the cytoplasm of motor neurons. Recent studies demonstrated that a significant percentage of familial ALS cases are caused by pathogenic mutations in the TAR DNA binding protein and fused in sarcoma/translocated in liposarcoma genes encoding, respectively, for TDP-43 and FUS proteins. Both TDP-43 and FUS are DNA/RNA-binding proteins involved in transcriptional regulation and splicing, shuttling, maturation and transport of mRNA molecules. Mutations in the two genes seem to induce a nucleo-cytoplasmic redistribution of FUS and TDP-43, possibly promoting aggregate formation and/or disrupting their physiological nuclear functions or their interactions with specific RNA targets. Those findings collectively suggest that alterations in cellular RNA metabolism may trigger motor neuron degeneration. PMID:20406182

Ticozzi, N; Ratti, A; Silani, V

2010-07-01

 
 
 
 
201

Protein aggregation and defective RNA metabolism as mechanisms for motor neuron damage.  

UK PubMed Central (United Kingdom)

The presence of protein inclusions within the central nervous system is a characteristic of most neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS). Aggregates may induce cell death trough several mechanisms, such as sequestration of essential cellular components, clogging of the proteasome system, and/or disruption of axonal transport. The neuropathological signature of ALS is represented by the presence of ubiquitinated inclusions immunoreactive for the protein TDP-43 in the cytoplasm of motor neurons. Recent studies demonstrated that a significant percentage of familial ALS cases are caused by pathogenic mutations in the TAR DNA binding protein and fused in sarcoma/translocated in liposarcoma genes encoding, respectively, for TDP-43 and FUS proteins. Both TDP-43 and FUS are DNA/RNA-binding proteins involved in transcriptional regulation and splicing, shuttling, maturation and transport of mRNA molecules. Mutations in the two genes seem to induce a nucleo-cytoplasmic redistribution of FUS and TDP-43, possibly promoting aggregate formation and/or disrupting their physiological nuclear functions or their interactions with specific RNA targets. Those findings collectively suggest that alterations in cellular RNA metabolism may trigger motor neuron degeneration.

Ticozzi N; Ratti A; Silani V

2010-07-01

202

Evaluation of nail abnormalities.  

UK PubMed Central (United Kingdom)

Knowledge of the anatomy and function of the nail apparatus is essential when performing the physical examination. Inspection may reveal localized nail abnormalities that should be treated, or may provide clues to an underlying systemic disease that requires further workup. Excessive keratinaceous material under the nail bed in a distal and lateral distribution should prompt an evaluation for onychomycosis. Onychomycosis may be diagnosed through potassium hydroxide examination of scrapings. If potassium hydroxide testing is negative for the condition, a nail culture or nail plate biopsy should be performed. A proliferating, erythematous, disruptive mass in the nail bed should be carefully evaluated for underlying squamous cell carcinoma. Longitudinal melanonychia (vertical nail bands) must be differentiated from subungual melanomas, which account for 50 percent of melanomas in persons with dark skin. Dystrophic longitudinal ridges and subungual hematomas are local conditions caused by trauma. Edema and erythema of the proximal and lateral nail folds are hallmark features of acute and chronic paronychia. Clubbing may suggest an underlying disease such as cirrhosis, chronic obstructive pulmonary disease, or celiac sprue. Koilonychia (spoon nail) is commonly associated with iron deficiency anemia. Splinter hemorrhages may herald endocarditis, although other causes should be considered. Beau lines can mark the onset of a severe underlying illness, whereas Muehrcke lines are associated with hypoalbuminemia. A pincer nail deformity is inherited or acquired and can be associated with beta-blocker use, psoriasis, onychomycosis, tumors of the nail apparatus, systemic lupus erythematosus, Kawasaki disease, and malignancy.

Tully AS; Trayes KP; Studdiford JS

2012-04-01

203

Evaluation of nail abnormalities.  

Science.gov (United States)

Knowledge of the anatomy and function of the nail apparatus is essential when performing the physical examination. Inspection may reveal localized nail abnormalities that should be treated, or may provide clues to an underlying systemic disease that requires further workup. Excessive keratinaceous material under the nail bed in a distal and lateral distribution should prompt an evaluation for onychomycosis. Onychomycosis may be diagnosed through potassium hydroxide examination of scrapings. If potassium hydroxide testing is negative for the condition, a nail culture or nail plate biopsy should be performed. A proliferating, erythematous, disruptive mass in the nail bed should be carefully evaluated for underlying squamous cell carcinoma. Longitudinal melanonychia (vertical nail bands) must be differentiated from subungual melanomas, which account for 50 percent of melanomas in persons with dark skin. Dystrophic longitudinal ridges and subungual hematomas are local conditions caused by trauma. Edema and erythema of the proximal and lateral nail folds are hallmark features of acute and chronic paronychia. Clubbing may suggest an underlying disease such as cirrhosis, chronic obstructive pulmonary disease, or celiac sprue. Koilonychia (spoon nail) is commonly associated with iron deficiency anemia. Splinter hemorrhages may herald endocarditis, although other causes should be considered. Beau lines can mark the onset of a severe underlying illness, whereas Muehrcke lines are associated with hypoalbuminemia. A pincer nail deformity is inherited or acquired and can be associated with beta-blocker use, psoriasis, onychomycosis, tumors of the nail apparatus, systemic lupus erythematosus, Kawasaki disease, and malignancy. PMID:22534387

Tully, Amber S; Trayes, Kathryn P; Studdiford, James S

2012-04-15

204

Mapping of alpha-neo-endorphin- and neurokinin B-immunoreactivity in the human brainstem.  

UK PubMed Central (United Kingdom)

We have studied the distribution of alpha-neo-endorphin- or neurokinin B-immunoreactive fibres and cell bodies in the adult human brainstem with no prior history of neurological or psychiatric disease. A low density of alpha-neo-endorphin-immunoreactive cell bodies was only observed in the medullary central gray matter and in the spinal trigeminal nucleus (gelatinosa part). Alpha-neo-endorphin-immunoreactive fibres were moderately distributed throughout the human brainstem. A high density of alpha-neo-endorphin-immunoreactive fibres was found only in the solitary nucleus (caudal part), in the spinal trigeminal nucleus (caudal part), and in the gelatinosa part of the latter nucleus. Neurokinin B-immunoreactive cell bodies (low density) were found in the periventricular central gray matter, the reticular formation of the pons and in the superior colliculus. The distribution of the neurokinin-immunoreactive fibres was restricted. In general, for both neuropeptides the density of the immunoreactive fibres was low. In the human brainstem, the proenkephalin system was more widely distributed than the prodynorphin system, and the preprotachykinin A system (neurokinin A) was more widely distributed than the preprotachykinin B system (neurokinin B).

Duque E; Mangas A; Salinas P; Díaz-Cabiale Z; Narváez JA; Coveńas R

2013-01-01

205

Myelin glycosphingolipid immunoreactivity and CSF levels in multiple sclerosis.  

UK PubMed Central (United Kingdom)

OBJECTIVES: Patients with multiple sclerosis were reported to harbour antibodies not only against proteins and glycoproteins but also against glycolipids, including sulfatide and galactosylceramide (GalCer), the two major glycosphingolipids of myelin. However, previous results were inconsistent concerning glycosphingolipid levels, antibody type, dominance of serum or Cerebrospinal fluid compartments and relationship to the multiple sclerosis (MS) course. RESULTS: We hereby report that the cerebrospinal fluid levels of sulfatide were increased in patients with MS (n = 46) compared with controls (n = 50, P < 0.001). In addition, patients had higher serum IgM anti-glycosphingolipid titres than controls (P = 0.03 for sulfatide, <0.001 for GalCer), while the anti-glycosphingolipid IgM antibodies in the cerebrospinal fluid were essentially normal. However, in seven of 46 patients cerebrospinal fluid IgG antibodies against GalCer (P = 0.004) could be detected, which was not found in any of the control individuals, and this finding might mirror the occurrence of more specific B-cell clones behind the blood-brain barrier. CONCLUSIONS: The IgM immunoreactivity in serum did not show any relationship to the type of course or severity of MS, arguing against a phenomenon secondary to myelin damage. Thus, the IgM antibody findings are compatible with an early antigen challenge or autoimmunity associated with natural antibodies.

Haghighi S; Lekman A; Nilsson S; Blomqvist M; Andersen O

2012-01-01

206

Deposition of immunoreactants in a cutaneous allergic drug reaction  

Directory of Open Access Journals (Sweden)

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; Billie L. Jackson; Michael S. Howard

2009-01-01

207

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

208

Sleep deprivation reduces neuroglobin immunoreactivity in the rat brain.  

UK PubMed Central (United Kingdom)

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.

Melgarejo-Gutiérrez M; Acosta-Peńa E; Venebra-Muńoz A; Escobar C; Santiago-García J; Garcia-Garcia F

2013-02-01

209

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

210

Immunoreactivity of normal and neoplastic human tissue mast cells.  

UK PubMed Central (United Kingdom)

Immunoreactivity of human tissue mast cells (TMCs) was studied in one case of solitary mastocytoma of the skin, three cases of malignant mastocytosis, and in six lymph nodes with reactive intrasinusoidal increase of TMCs. Immunohistochemically, TMCs reacted positively to antisera against vimentin, common leukocyte antigen (CLA), lysozyme, alpha 1-antitrypsin (alpha 1-AT), and alpha 1-antichymotrypsin (alpha 1-ACT) and to a monoclonal antibody (KiB3) that detects preferentially B-lymphocytes. Additionally, strong positive reactions to polyclonal antisera against adrenocorticotropic hormone (ACTH) and human peptide histidine isoleucine (PHI) and weaker reactions to antisera against leu-enkephalin and met-enkephalin were observed; all other antisera tested yielded negative results. Positive stainings for vimentin, CLA, alpha 1-AT, alpha 1-ACT, and lysozyme further support the hypothesis that human TMCs may be related to the myeloid-monocytic system. The positive reactivity of TMCs to antisera against ACTH, PHI, leu-enkephalin, and met-enkephalin has not been reported previously. These findings suggest that TMCs are able to store and/or produce regulatory peptides in addition to many other well-known, granule-bound mediators.

Horny HP; Reimann O; Kaiserling E

1988-03-01

211

Systemic abnormalities in liver disease  

Directory of Open Access Journals (Sweden)

Full Text Available Systemic abnormalities often occur in patients with liver disease. In particular, cardiopulmonary or renal diseases accompanied by advanced liver disease can be serious and may determine the quality of life and prognosis of patients. Therefore, both hepatologists and non-hepatologists should pay attention to such abnormalities in the management of patients with liver diseases.

Masami Minemura, Kazuto Tajiri, Yukihiro Shimizu

2009-01-01

212

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

213

Calcitonin gene-related peptide immunoreactivity in prurigo nodularis: a comparative study with neurodermatitis circumscripta.  

UK PubMed Central (United Kingdom)

Eight patients with intensely pruritic lesions of chronic idiopathic prurigo nodularis and three patients with neurodermatitis circumscripta were investigated using the indirect immunofluorescence method. Results showed similarities in epidermal hyperplasia but not in nerve proliferation and neuropeptide immunoreactivity. Increased numbers of calcitonin gene-related peptide (CGRP) and substance P immunoreactive nerve fibre bundles were detected in specimens taken from prurigo nodularis lesions, but no increased immunoreactivity could be seen in specimens taken from patients having neurodermatitis circumscripta compared to normal skin. The neuropeptides, CGRP and substance P, may be responsible for the intense itching of prurigo nodularis lesions.

Vaalasti A; Suomalainen H; Rechardt L

1989-05-01

214

Distribution and chromatographic characterization of peptide tyrosine leucine amide (PYLa) immunoreactivity in mammalian tissues.  

UK PubMed Central (United Kingdom)

Using a polyclonal antiserum raised against the mid-region of the abundant amphibian skin peptide PYLa (peptide tyrosine leucine amide) we have shown that immunoreactive PYLa is present in a wide variety of rat tissues, being particularly abundant in the liver, spleen and gut. In each tissue the predominant molecular form corresponds to a moderately hydrophobic, basic peptide of about 10 amino acids. Using the same antiserum in immunohistochemistry we have demonstrated that immunoreactive PYLa was present in endocrine cells, but not neurons, in the ileal mucosa of rat and man. These findings are consistent with an hormonal or local tissue regulatory role for immunoreactive PYLa in the intestine.

Giraud AS; Furness JB; Parker L

1991-01-01

215

Distribution and chromatographic characterization of peptide tyrosine leucine amide (PYLa) immunoreactivity in mammalian tissues.  

Science.gov (United States)

Using a polyclonal antiserum raised against the mid-region of the abundant amphibian skin peptide PYLa (peptide tyrosine leucine amide) we have shown that immunoreactive PYLa is present in a wide variety of rat tissues, being particularly abundant in the liver, spleen and gut. In each tissue the predominant molecular form corresponds to a moderately hydrophobic, basic peptide of about 10 amino acids. Using the same antiserum in immunohistochemistry we have demonstrated that immunoreactive PYLa was present in endocrine cells, but not neurons, in the ileal mucosa of rat and man. These findings are consistent with an hormonal or local tissue regulatory role for immunoreactive PYLa in the intestine. PMID:2052495

Giraud, A S; Furness, J B; Parker, L

216

Immunoreactive oxytocin and vasopressin in the non-pregnant human uterus and oviductal isthmus  

DEFF Research Database (Denmark)

The regional distribution of immunoreactive OT and AVP in the human uterus was investigated. Specimens of non-pregnant human uterus and oviduct were homogenized and extracted. The tissue levels exceeded the plasma concentrations of the peptides. The largest quantities of both peptides were found in the cervix and oviductal isthmus. The amounts found in the uterine fundus and isthmus were, however, not significantly different. Only 23% of immunoreactive OT eluted in the position of standard peptide on high-performance liquid chromatography. All immunoreactive AVP eluted with standard AVP after additional ether extraction of octadecasilyl extracts. We conclude that the human uterus contains materials immunologically and chromatographically identical to oxytocin and vasopressin.

Lundin, S; Forman, Axel

1989-01-01

217

Demonstration of GABA immunoreactive cells in the inferior olive of baboons (Papio papio and Papio anubis).  

UK PubMed Central (United Kingdom)

The distribution of gamma-aminobutyric acid (GABA)-like immunoreactivity was studied in semithin sections through the inferior olivary complex in two baboon species. About 5% of the olivary neurons were GABA-immunoreactive. The GABA-immunoreactive neurons differed from the large majority of olivary neurons by their smaller size and their lower contents of aspartate, as judged by analysis of alternate sections labelled with an aspartate antiserum. The present observations raise the possibility that in primates the GABAergic modulation of the activity of the climbing fibre system is effected not only by the previously described input from the cerebellar nuclei and other extrinsic sources, but that intrinsic neurons also participate.

Walberg F; Ottersen OP

1989-06-01

218

Calretinin immunoreactivity in the developing olfactory system of the rainbow trout  

Digital Repository Infrastructure Vision for European Research (DRIVER)

olfactory system of the rainbow trout was studied by using an indirect immunocytochemical method. Calretinin immunoreactivity was firstly detected at 150 day-degrees in the olfactory placode, where labeled primordial cells were observ...

Porteros Herrero, Ángel; Arévalo Arévalo, Rosario; Weruaga, Eduardo; Crespo, Carlos; Brińón, Jesús G.; Alonso Peńa, José-Ramón

219

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

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 immunoflu...

Cole, G T; Seshan, K R; Franco, M; Bukownik, E; Sun, S H; Hearn, V M

220

Intense cytoplasmic ezrin immunoreactivity predicts poor survival in colorectal cancer.  

Science.gov (United States)

Ezrin is a membrane-cytoskeleton anchor, which, in experimental models, regulates tumor cell invasion and metastatic ability. We carried out immunohistochemical analysis of ezrin in 74 advanced colorectal cancer patients and correlated it to clinicopathologic variables and disease outcome. In contrast to the predominantly membraneous immunoreactivity of normal colorectal epithelium, ezrin expression in the colorectal cells was typically cytoplasmic. Altogether, 16.2% (12/74) of the tumors showed negative/weak ezrin staining, 35.1% (26/74) had moderate staining, and 48.6% (36/74) had intense staining. The expression was more intense in colon than in rectal carcinomas (P = .003). Increased ezrin expression was associated with adverse outcome, that is, shorter disease-specific survival; 48.3 months and 36.6 months for negative-weak versus intense expression (P = .041) as well as shorter survival with metastases at 36 months (P = .030); the metastases(36) rates in ezrin(neg/weak), ezrin(moderate), ezrin(intense) are 58.3%, 25.0%, and 18.4%, respectively. In univariate survival analysis, dichotomized (negative/weak versus moderate/strong) ezrin expression significantly predicted both the 5-year disease specific survival (P = .035) and 5-year metastases (P = .018) but lost this predictive power in multivariate (Cox) analysis. High ezrin expression was also related to high E-cadherin (cytoplasmic) expression, DNA aneuploidy, and high thymidylate synthase expression (P = .046, P = .042, P = .046, respectively). These results suggest that ezrin may play a role in colorectal cancer progression and that ezrin expression might provide clinically valuable information in predicting the biological behavior of colorectal cancer. PMID:18701134

Elzagheid, Adam; Korkeila, Eija; Bendardaf, Riyad; Buhmeida, Abdelbaset; Heikkilä, Suvi; Vaheri, Antti; Syrjänen, Kari; Pyrhönen, Seppo; Carpén, Olli

2008-08-12

 
 
 
 
221

Comparison of immunoreactive serum trypsinogen and lipase in Cystic Fibrosis  

International Nuclear Information System (INIS)

[en] 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

1984-01-01

222

Comparison of immunoreactive serum trypsinogen and lipase in Cystic Fibrosis  

Energy Technology Data Exchange (ETDEWEB)

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.

Lloyd-Still, J.D.; Weiss, S.; Wessel, H.; Fong, L.; Conway, J.J.

1984-01-01

223

Jejunum abnormalities at MR enteroclysis  

International Nuclear Information System (INIS)

Objective: MR enteroclysis has become an important tool to visualize the complete small bowel wall and extramural structures. In many centers, this technique is rapidly becoming the first-line technique for small bowel visualization. MR enteroclysis yields a diagnosis of thickened jejunal loops in some patients. In this paper, we describe an MR enteroclysis protocol and review the literature on jejunum abnormalities with several sample cases. Conclusion: Jejunum abnormalities are not uncommon. These abnormalities can be self-limiting, but some patients suffer from infectious and other pathologic conditions of the small bowel necessitating intervention.

2008-01-01

224

Jejunum abnormalities at MR enteroclysis  

Energy Technology Data Exchange (ETDEWEB)

Objective: MR enteroclysis has become an important tool to visualize the complete small bowel wall and extramural structures. In many centers, this technique is rapidly becoming the first-line technique for small bowel visualization. MR enteroclysis yields a diagnosis of thickened jejunal loops in some patients. In this paper, we describe an MR enteroclysis protocol and review the literature on jejunum abnormalities with several sample cases. Conclusion: Jejunum abnormalities are not uncommon. These abnormalities can be self-limiting, but some patients suffer from infectious and other pathologic conditions of the small bowel necessitating intervention.

Wiarda, Bart M. [Department of Radiology, Medical Center Alkmaar, Wilhelminalaan 12, 1815 JD Alkmaar (Netherlands)], E-mail: b.m.wiarda@mca.nl; Heine, Dimitri G.N. [Department of Gastroenterology, Medical Center Alkmaar (Netherlands); Rombouts, Marieke C. [Department of Radiology, Bronovo Hospital, P.O. Box 96900, NL-2509 JH The Hague (Netherlands); Kuipers, Ernst J. [Department of Gastroenterology and Hepatology, Erasmus University Medical Center, P.O. Box 2400, NL-3000 CA Rotterdam (Netherlands); Stoker, Jaap [Department of Radiology, Academic Medical Center Amsterdam, P.O. Box 22660, NL-1100 DD (Netherlands)

2008-07-15

225

A simple method to determine the immunoreactivity of radiolabelled monoclonal antibodies to the TAG-72 antigen  

International Nuclear Information System (INIS)

A simple method has been developed for determining the immunoreactivity of radiolabelled monoclonal antibodies to the TAG-72 antigen. The method involves binding of a constant small amount of the antibody to increasing concentrations of bovine submaxillary mucin. The immunoreactive fraction (IRF) is then determined by linear extrapolation of binding to infinite antigen excess. Using this assay, the IRF of radioiodinated anti-TAG-72 antibodies ranged from 0.22-0.48. (author).

1993-01-01

226

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

227

Characteristics of galanin and vasoactive intestinal peptide immunoreactivity in the rat amygdala complex  

Directory of Open Access Journals (Sweden)

Full Text Available Introduction Morphological features and morphometric parameters of galanin (GAL) and vasoactive intestinal peptide (VIP) immunoreactive neurons and neuronal fibres were studied in all nuclei of adult male rat amygdala. Material and methods After perfusion and fixation, rat brains were immunohistochemically stained with antibodies against GAL and VIP and then visualized by avidin-biotin-peroxidase complex. Results and Discussion The greatest number of galanin-immunoreactive neurons were identified in the medial part of the central nucleus and in the dorsal part of the medial nucleus. In the first case, most neurons were bipolar (37%), and in the second, they were ovoid (45%). GAL-immunoreactive fibers were identified in the medial nucleus, "bed nucleus" of the accessory olfactory tract, frontal cortical nucleus, amygdalo-hippocampal area and basolateral nucleus. VIP-immunoreactive neurons were diffusely distributed in more nuclei than the previous, mostly in the lateral, basolateral, and basomedial nucleus. They were mostly ovoid (40%). VIP-immunoreactive fibers were observed in the lateral part of the central nucleus, while long and radially oriented fibers were present in the frontal and dorsal cortical nucleus. Conclusion By distribution analysis of GAL and VIP immunoreactive neurons and fibers, and according to literature data, it can be assumed that the medial part of the central nucleus receives VIP fibers from other parts of the amygdaloid body, and then sends GAL fibers to the medial nucleus.

Puškaš Laslo; Puškaš Nela; Malobabi? Slobodan; Krivoku?a Dragan; Stankovi? Gordana; Radonji? Vidosava

2007-01-01

228

Detection of Abnormalities in MANETs.  

Science.gov (United States)

Abnormalities in MANETs can be malicious attacks or selfish nodes which can affect network architecture and network operation significantly. Clearly, there are two distinct objectives: 1) To design/examine attacks and develop countermeasures and 2) design...

W. Wang

2007-01-01

229

TMI abnormal waste project plan  

Energy Technology Data Exchange (ETDEWEB)

This report discusses plans for the TMI Abnormal Waste Project, which is part of the EPICOR and Waste Research and Disposition Program and funded by the US Department of Energy. The sequence proposed for disposition of Three Mile Island (TMI) abnormal wastes includes: (a) packaging at TMI, (b) shipment to the Idaho National Engineering Laboratory (INEL), (c) storage at INEL for up to 30 years, (d) processing for disposal, and (e) final disposal. Some wastes may be disposable immediately and would be processed and disposed without storage. Potentially, 930 ft/sup 3/ of cartridge filters, Submerged Demineralizer System filters, sludges, ion-exchange resins, and miscellaneous plant equipment may be classified as abnormal waste. Some wastes may be deleted and others added as cleanup progresses at TMI. The first waste classified as abnormal is Makeup and Purification Demineralizer resin. This report outlines storage plans, procedures, project management, costs, and schedules for placement of those resins.

Ayers, A.L. Jr.

1984-06-01

230

Chromosomal abnormalities in B-CLL.  

UK PubMed Central (United Kingdom)

Amongst 141 patients with B-CLL, 53% had an abnormal karyotype. Treatment free survival was shorter in those with karyotypic abnormalities, and especially in those with multiple abnormalities. Multiple abnormalities tended to be associated with progressive disease. Trisomy 12 alone carried no worse a prognosis than a normal karyotype. 49% of patients with stage AO disease had an abnormal karyotype.

Oscier DG; Fitchett M; Hamblin TJ

1988-01-01

231

Development of a canine trypsin-like immunoreactivity assay system using monoclonal antibodies.  

Science.gov (United States)

The radioimmunoassay (RIA) for trypsin-like immunoreactivity (TLI) is one of the most sensitive and specific tests for detecting exocrine pancreatic insufficiency (EPI). An abnormally low serum TLI concentration (<2.5 ng/ml) indicates end-stage EPI. Although RIA methods can be used to detect canine serum TLI, these procedures are beyond the capabilities of most veterinary clinics and general laboratories. Using monoclonal antibodies (mAbs), we developed an enzyme-linked immunosorbent assay (ELISA) for canine TLI and incorporated it into an immunochromatographic test (ICT) for the diagnosis of EPI. The ELISA was linear over TLI concentrations of 1-100 ng/ml. Levels of intra-assay coefficients of variance (CVs) were 1.8-6.1%, inter-assay CVs were 5.1-9.8%, and the recovery of TLI added to two samples of canine serum ranged from 89 to 111 and 93 to 108%, respectively. Good correlation (correlation coefficient, 0.974) occurred between the TLI values obtained by the ELISA method and those by RIA from 56 clinical samples. Serum TLI values in clinically healthy dogs ranged from 7.8 to 29.2 ng/ml by ELISA, and those from dogs with EPI were 0.0-0.6 ng/ml. The values were 0.0-287.4 ng/ml for dogs with pancreatitis, and those from dogs with gastrointestinal disease were 5.5-58.9 ng/ml. The only statistically significant difference (P<0.01) occurred between the TLI level of healthy dogs and those with EPI. The ICT kit showed high reproducibility, and the TLI values yielding negative results differed significantly (P<0.01) from those returning positive results. The ICT kit yielded negative results (indicating EPI) from clinical serum samples with TLI concentrations of 0.0-4.1 ng/ml by ELISA. Both the ELISA and ICT kit are useful tools in the diagnosis of canine EPI. PMID:12052341

Waritani, Takaki; Okuno, Yoko; Ashida, Yoshinori; Hisasue, Masaharu; Tsuchiya, Ryo; Kobayashi, Kosaku; Yamada, Takatsugu

2002-08-01

232

SPERM ABNORMALITIES AND ITS TREATMENT  

Directory of Open Access Journals (Sweden)

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.; Solanki Anil S.

2011-01-01

233

Ciliary abnormalities in respiratory disease.  

Science.gov (United States)

One hundred and sixty seven children, ranging in age from 5 weeks to 16 years, with chronic upper or lower respiratory tract problems, or both, were investigated for ciliary dyskinesia. Abnormal ciliary function was found in 18 cases all of whom had chronic lower respiratory disease and most of whom also had upper respiratory problems. Fifteen of the 18 cases had reduced ciliary beat frequencies (less than 10 Hz) associated with dyskinesia and the other three showed apparent absence of ciliated cells. Of the 15 cases with reduced ciliary beat frequencies, ciliary ultrastructure was normal in seven cases but abnormal with missing dynein arms and occasional abnormalities of microtubular arrangement in eight. Respiratory symptoms in the perinatal period were more common in children with abnormal ciliary function and present in all those with ultrastructural abnormalities or absence of ciliated cells compared with 34 (26%) of 132 children, in whom symptoms were recorded, with normal ciliary function. This study would suggest that all children with unexplained chronic respiratory disease, in particular those with symptoms starting in the perinatal period, should be investigated for ciliary dyskinesia. PMID:3355203

Buchdahl, R M; Reiser, J; Ingram, D; Rutman, A; Cole, P J; Warner, J O

1988-03-01

234

Ciliary abnormalities in respiratory disease.  

UK PubMed Central (United Kingdom)

One hundred and sixty seven children, ranging in age from 5 weeks to 16 years, with chronic upper or lower respiratory tract problems, or both, were investigated for ciliary dyskinesia. Abnormal ciliary function was found in 18 cases all of whom had chronic lower respiratory disease and most of whom also had upper respiratory problems. Fifteen of the 18 cases had reduced ciliary beat frequencies (less than 10 Hz) associated with dyskinesia and the other three showed apparent absence of ciliated cells. Of the 15 cases with reduced ciliary beat frequencies, ciliary ultrastructure was normal in seven cases but abnormal with missing dynein arms and occasional abnormalities of microtubular arrangement in eight. Respiratory symptoms in the perinatal period were more common in children with abnormal ciliary function and present in all those with ultrastructural abnormalities or absence of ciliated cells compared with 34 (26%) of 132 children, in whom symptoms were recorded, with normal ciliary function. This study would suggest that all children with unexplained chronic respiratory disease, in particular those with symptoms starting in the perinatal period, should be investigated for ciliary dyskinesia.

Buchdahl RM; Reiser J; Ingram D; Rutman A; Cole PJ; Warner JO

1988-03-01

235

Differences between unipolar and bipolar I depression in the quantitative analysis of glutamic acid decarboxylase-immunoreactive neuropil.  

UK PubMed Central (United Kingdom)

Alterations in GABAergic neurotransmission are assumed to play a crucial role in the pathophysiology of mood disorders. Glutamic acid decarboxylase (GAD) is the key enzyme in GABA synthesis. This study aimed to differentiate between unipolar and bipolar I depression using quantitative evaluation of GAD-immunoreactive (GAD-ir) neuropil in several brain regions known to be involved in the pathophysiology of mood disorders. Immunohistochemical staining of GAD 65/67 was performed in the orbitofrontal, anterior cingulate and dorsolateral prefrontal cortex (DLPFC), the entorhinal cortex, the hippocampal formation and the medial dorsal and lateral dorsal (LD) thalamic nuclei, with a quantitative densitometric analysis of GAD-ir neuropil. The study was performed on paraffin-embedded brains from 9 unipolar and 12 bipolar I depressed patients (8 and 6 suicidal patients, respectively) and 18 matched controls. In unipolar patients, compared with controls, only the increased relative density of GAD-ir neuropil in the right LD was different from the previous results in depressed suicides from the same cohort (Gos et al. in J Affect Disord 113:45-55, 2009). On the other hand, the left DLPFC was the only area where a significant decrease was observed, specific for bipolar I depression. Significant differences between both diagnostic groups were found in these regions. By revealing abnormalities in the relative density of GAD-ir neuropil in brain structures, our study suggests a diathesis of the GABAergic system in mood disorders, which may differentiate the pathophysiology of unipolar from that of bipolar I depression.

Gos T; Steiner J; Bielau H; Dobrowolny H; Günther K; Mawrin C; Krzy?anowski M; Hauser R; Brisch R; Bernstein HG; Jankowski Z; Braun K; Bogerts B

2012-12-01

236

Distribution of APGWamide-immunoreactivity in the brain and reproductive organs of adult pygmy squid, Idiosepius pygmaeus.  

Science.gov (United States)

The neuropeptide APGWamide is involved in the control of the reproductive behavior in molluscs. Using immunocytochemistry, we investigated the distribution of APGWa-immunoreactive neurons in the brain and reproductive organs of adult male and female specimen of Idiosepius pygmaeus. The study showed that the APGWamide-immunoreactive neurons and fibers are localized in the dorsal basal and vertical lobes of the supraesophageal mass, the palliovisceral lobe of the posterior subesophageal mass and olfactory lobe of the optic tract in male brains, with the highest number of APGWamide-immunoreactive neurons in the palliovisceral and olfactory lobes. In females, only the palliovisceral and olfactory lobes contained APGWa-immunoreactive neurons. The number of APGWamide-immunoreactive neurons in male I. pygmaeus brain is significantly higher than in females. Furthermore, APGWamide-immunoreactive fibers are localized exclusively in male reproductive organs and mantle muscles. Together these data suggest a role for APGW-amide in the control of male reproduction. PMID:21915688

Sirinupong, P; Suwanjarat, J; van Minnen, J

2011-09-14

237

Distribution of APGWamide-immunoreactivity in the brain and reproductive organs of adult pygmy squid, Idiosepius pygmaeus.  

UK PubMed Central (United Kingdom)

The neuropeptide APGWamide is involved in the control of the reproductive behavior in molluscs. Using immunocytochemistry, we investigated the distribution of APGWa-immunoreactive neurons in the brain and reproductive organs of adult male and female specimen of Idiosepius pygmaeus. The study showed that the APGWamide-immunoreactive neurons and fibers are localized in the dorsal basal and vertical lobes of the supraesophageal mass, the palliovisceral lobe of the posterior subesophageal mass and olfactory lobe of the optic tract in male brains, with the highest number of APGWamide-immunoreactive neurons in the palliovisceral and olfactory lobes. In females, only the palliovisceral and olfactory lobes contained APGWa-immunoreactive neurons. The number of APGWamide-immunoreactive neurons in male I. pygmaeus brain is significantly higher than in females. Furthermore, APGWamide-immunoreactive fibers are localized exclusively in male reproductive organs and mantle muscles. Together these data suggest a role for APGW-amide in the control of male reproduction.

Sirinupong P; Suwanjarat J; van Minnen J

2011-12-01

238

Electrocardiographic abnormalities in neurological diseases  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english OBJECTIVE: To evaluate electrocardiographic abnormalities in patients with neurologic diseases. METHODS: We studied 161 patients with neurologic disorders by analyzing the 12-lead electrocardiogram during the pathological process. An expert who did not know anything about the patients evaluated the traces. RESULTS: Neurological process included brain tumor (41%), stroke (27.3%), cerebral aneurysm (15.5%), subarachnoid hemorrhage (6.8%), subdural hemorrhage (5%), and head (more) injury (4.4%). Electrocardiograms were normal in 61% of cases, and the most frequent abnormality was ventricular repolarization (23.7%). The presence of T waves (4.6%) and prolonged QT intervals (8.8%) was the most characteristic of brain injuries. CONCLUSION: We observed a lower incidence of electrocardiographic abnormalities than that described in the literature.

Póvoa, Rui; Cavichio, Luciano; Almeida, Ana Lúcia de; Viotti, Danielle; Ferreira, Celso; Galvăo, Luciane; Pimenta, Joăo

2003-04-01

239

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

240

Serotonin-immunoreactive sensory neurons in the antenna of the cockroach Periplaneta americana.  

UK PubMed Central (United Kingdom)

Antennae of insects contain of a vast array of sensory neurons that process olfactory, gustatory, mechanosensory, hygrosensory and thermosensory information. Except those with multimodal functions, most sensory neurons use acetylcholine as a neurotransmitter. Using immunohistochemistry combined with retrograde staining of antennal sensory neurons in the cockroach Periplaneta americana, we found serotonin-immunoreactive sensory neurons in the antenna. These were selectively distributed in chaetic and scolopidial sensilla, and in the scape, the pedicel and first 15 segments of the flagellum. A single serotonin-immunoreactive sensory neuron co-habited a chaetic sensillum with up to four serotonin-negative sensory neurons. Based on their morphological features, serotonin-immunopositive and -negative sensory neurons might process mechanosensory and contact chemosensory modalities, respectively. Scolopidial sensilla constitute the chordotonal and Johnston's organs within the pedicel, and process antennal vibrations. Immuno-electron microscopy clearly revealed that serotonin-immunoreactivities selectively localize to a specific type of mechanosensory neuron, called type 1 sensory neuron. In a chordotonal scolopidial sensillum, a serotonin-immunoreactive type 1 neuron always paired with a serotonin-negative type 1 neuron. Conversely, serotonin-immunopositive and -negative type 1 neurons were randomly distributed in Johnston's organ. In the deutocerebrum, serotonin-immunoreactive sensory neuron axons formed three different sensory tracts and those from distinct types of sensilla terminated in distinct brain regions. Our findings indicate that a biogenic amine, serotonin, may act as a neurotransmitter in peripheral mechanosensory neurons. J. Comp. Neurol., 2013. © 2013 Wiley Periodicals, Inc.

Watanabe H; Shimohigashi M; Yokohari F

2013-07-01

 
 
 
 
241

Urocortin-like immunoreactivity in the primary lymphoid organs of the duck (Anas platyrhynchos)  

Directory of Open Access Journals (Sweden)

Full Text Available Urocortin (UCN) is a 40 aminoacid peptide which belongs to corticotropin-releasing factor (CRF) family. This family of peptides stimulates the secretion of proopiomelanocortin (POMC)-derived peptides, adrenocorticotropic hormone (ACTH), b-endorphin and melanocyte-stimulating hormone (MSH) in the pituitary gland. In the present study, using Western blotting and immunohistochemistry, the distribution of UCN in the primary lymphoid organs of the duck was investigated at different ages. In the cloacal burse and thymus, Western blot demonstrated the presence of a peptide having a molecular weight compatible with that of the mammalian UCN. In the cloacal burse, immunoreactivity was located in the medullary epithelial cells and in the follicular associated and cortico-medullary epithelium. In the thymus, immunoreactivity was located in single epithelial cells. Double labelling immunofluorescence studies showed that UCN immunoreactivity completely colocalised with cytokeratin immunoreactivity in both the thymus and cloacal burse. Statistically significant differences in the percentage of UCN immunoreactivity were observed between different age periods in the cloacal burse. The results suggest that, in birds, urocortin has an important role in regulating the function of the immune system.

A. De Luca; C. Squillacioti; M.E. Pero; S. Paino; E. Langella; N. Mirabella

2009-01-01

242

FMRFamide immunoreactivity in the nervous system of the medusa Polyorchis penicillatus  

DEFF Research Database (Denmark)

Three different antisera to the molluscan neuropeptide Phe-Met-Arg-Phe-amide (FMRFamide) and two different antisera to the fragment RFamide were used to stain sections or whole mounts of the hydrozoan medusa Polyorchis penicillatus. All antisera stained the same neuronal structures. Strong immunoreactivity was found in neurons of the ectodermal nerve nets of the manubrium and tentacles, in neurons of the sensory epithelium, and in neurons at the periphery of the sphincter muscle. Strong immunoreactivity was also present in processes and perikarya of the whole outer nerve ring, in the ocellar nerves, and in nerve cells lying at the periphery of the ocellus. The inner nerve ring contained a moderate number of immunoreactive processes and perikarya, which were distinct from the swimming motor neurons. In contrast to the situation in the hydrozoan polyp Hydra attenuata, no immunoreactivity was found with several antisera to oxytocin/vasopressin and bombesin/gastrin-releasing peptide. The morphology and location of most FMRFamide-immunoreactive neurons in Polyorchis coincides with two identified neuronal systems, which have been recently discovered from neurophysiological studies.

Grimmelikhuijzen, C J; Spencer, A N

1984-01-01

243

Hereditary tyrosinemia type I: lack of correlation between clinical findings and amount of immunoreactive fumarylacetoacetase protein.  

UK PubMed Central (United Kingdom)

Immunoblot analyses with bovine fumarylacetoacetase antibodies have been performed in fibroblast extracts from 28 patients with hereditary tyrosinemia of various clinical phenotypes, in one healthy individual homozygous for a "pseudodeficiency" gene for fumarylacetoacetase, and in three tyrosinemia families in which one or both parents are compound heterozygotes for the tyrosinemia and pseudodeficiency genes. Liver extracts from two chronic patients were also investigated. None of the patients with the acute type of tyrosinemia had detectable immunoreactive protein in fibroblast extracts. Only two of seven patients with typical chronic tyrosinemia had definite immunoreactivity in fibroblasts. In liver tissue, one of the patients had cross-reactive material and the other had no immunoreactivity. Four of 13 patients with intermediate clinical findings showed immunoreactivity in fibroblasts. There was no relationship between severity of symptoms and amount of cross-reactive material in this group. The pseudodeficiency gene product gave almost no detectable immunoreactivity in fibroblasts. The results indicate that chronic tyrosinemia may be due to at least two protein variants, and immunoblotting does not classify tyrosinemia patients according to clinical findings.

Kvittingen EA; Rootwelt H; van Dam T; van Faassen H; Berger R

1992-01-01

244

Comparative measurements of immunoreactive renin, plasma renin activity and angiotensin II in human plasma.  

UK PubMed Central (United Kingdom)

In human plasma samples we compared the values of renin activity, determined with a conventional enzymatic assay, with those of immunoreactive renin, determined with a new, direct immunoradiometric assay which employs highly specific monoclonal antibodies, and with those of angiotensin II; the comparative measurements of renin were carried out also in trypsin activated samples of nephric and anephric subjects. We found that, overall, there was a close relationship between renin activity and immunoreactive renin; however, this relationship was absent when the statistical analysis was restricted to plasmas with low or very low renin. We also found that, within a rather wide range of values, angiotensin II was more closely correlated with immunoreactive renin than with renin activity. Trypsin activation increased to a similar extent immunoreactive renin and renin activity in plasma of nephric and anephric subjects and, overall, the values of total renin obtained with the two assays were significantly correlated. The results of these comparative determinations indicate that, in general, the measurement of immunoreactive renin represents a valid alternative to that of renin activity and a reliable index of the activity of the renin-angiotensin system. In addition, studies with trypsin activation suggest that even in the anephric state human plasma contains an inactive enzyme convertible into an active form which has immunological properties similar to those of active renin.

Morganti A; Turolo L; Pulazzini E; Zanchetti A

1987-01-01

245

Comparative measurements of immunoreactive renin, plasma renin activity and angiotensin II in human plasma.  

Science.gov (United States)

In human plasma samples we compared the values of renin activity, determined with a conventional enzymatic assay, with those of immunoreactive renin, determined with a new, direct immunoradiometric assay which employs highly specific monoclonal antibodies, and with those of angiotensin II; the comparative measurements of renin were carried out also in trypsin activated samples of nephric and anephric subjects. We found that, overall, there was a close relationship between renin activity and immunoreactive renin; however, this relationship was absent when the statistical analysis was restricted to plasmas with low or very low renin. We also found that, within a rather wide range of values, angiotensin II was more closely correlated with immunoreactive renin than with renin activity. Trypsin activation increased to a similar extent immunoreactive renin and renin activity in plasma of nephric and anephric subjects and, overall, the values of total renin obtained with the two assays were significantly correlated. The results of these comparative determinations indicate that, in general, the measurement of immunoreactive renin represents a valid alternative to that of renin activity and a reliable index of the activity of the renin-angiotensin system. In addition, studies with trypsin activation suggest that even in the anephric state human plasma contains an inactive enzyme convertible into an active form which has immunological properties similar to those of active renin. PMID:3308196

Morganti, A; Turolo, L; Pulazzini, E; Zanchetti, A

1987-01-01

246

CXCL14-like immunoreactivity in growth hormone-containing cells of urodele pituitaries.  

UK PubMed Central (United Kingdom)

Immunohistochemical techniques were employed to investigate the distribution of a chemokine, namely, CXCL14-like immunoreactivity in the axolotl (Ambystoma mexicanum) and Japanese black salamander (Hynobius nigrescens) pituitaries. CXCL14-immunoreactive cells concentrated at an area of the pars distalis adjacent to the pars intermedia. We found that these cells correspond to the cells immunoreactive to an antibody against rat growth hormone (GH). Immunoelectron microscopy indicated that the CXCL14-like substance and GH coexisted on the secretory granules in the axolotl pituitary. Western blot analysis of axolotl pituitary extracts revealed the anti-human CXCL14 antibody labeled an approximately 16.6-kDa band that was not labeled by the anti-GH antibody. The CXCL14-like substance in the pars distalis may participate in GH functions in these species.

Suzuki H; Itoh M; Yamamoto T

2012-02-01

247

Galanin immunoreactivity in the brain of the desert lizard Uromastyx acanthinura during activity season.  

UK PubMed Central (United Kingdom)

The distribution of galanin immunoreactive perikarya and nerve fibers in the brain of the desert lizard U. acanthinura was studied by means of immunofluorescence using an antiserum against rabbit galanin. The animals were captured during the activity season in March (wet season) just before reproduction period and in June (arid season) after ovulation period. Immunoreactive neurons were mostly detected in the mediobasal and the infundibular recess nuclei, the nucleus of the paraventricular organ, the paraventricular organ, the periventricular nucleus and in the anterior hypothalamus at the level of the periventricular nucleus, the paraventricular nucleus and the supraoptic nucleus. The differences in brain galanin expression between animals collected under both sets of environmental conditions indicated changes which occur during the annual and reproductive cycles. The wide hypothalamic and extrahypothalamic distribution of galanin immunoreactive fibers suggests that this peptide may have hypophysiotropic, neuromodulator and neurotransmitter roles in the lizard U. acanthinura.

Hammouche SB; Bennis M

2013-01-01

248

Drebrin immunoreactivity in the striatum of a rat model of levodopa-induced dyskinesia.  

UK PubMed Central (United Kingdom)

Levodopa-induced dyskinesia has been suggested to result from maladaptive plasticity at corticostriatal synapses. Synaptic plasticity is based upon morphologic changes of dendritic spines. To elucidate whether the morphologic changes of spines occur in the striatum of rat models of levodopa-induced dyskinesia, we examined immunoreactivity of drebrin, an actin-binding protein localized in dendritic spines of excitatory synapses, using 6-hydroxydopamine-lesioned rats repeatedly treated with levodopa. The cross-sectional area of drebrin-immunoreactive organelles, putative spines, in the dopamine-denervated striatum of the levodopa-induced dyskinesia model was greater than that of the Parkinson's disease model. Immunoelectron microscopic examinations confirmed that drebrin-immunoreactive spines became enlarged in the dopamine-denervated striatum of the levodopa-induced dyskinesia model, but not in the Parkinson's disease model. These results suggest that the development of levodopa-induced dyskinesia is associated with enlargement of dendritic spines at corticostriatal excitatory synapses.

Nishijima H; Arai A; Kimura T; Mori F; Yamada J; Migita K; Wakabayashi K; Baba M; Ueno S; Tomiyama M

2013-08-01

249

beta-Casomorphin immunoreactive materials in cows' milk incubated with various bacterial species.  

UK PubMed Central (United Kingdom)

To obtain information about the possible release of beta-casomorphins from beta-casein under in vitro conditions, cows' milk was incubated with 13 strains of Gram-negative or Gram-positive bacterial species isolated from bovine milk. After incubation periods of 1-24 d, milk samples were assayed for beta-casomorphin-4, -5, -6 or -7 immunoreactive materials. In general, no beta-casomorphin immunoreactive material was found in samples incubated with non-caseolytic strains, e.g. Pseudomonas putida or Streptococcus faecalis subsp. faecalis; however, high levels were found in several samples incubated with caseolytic strains, e.g. Ps. aeruginosa or Bacillus cereus. When such materials were present, the amounts of beta-casomorphin-4, -6 or -7 greatly exceeded beta-casomorphin-5 immunoreactivity. It was concluded that while some of these materials probably represented authentic beta-casomorphins, others were more likely to be beta-casomorphin precursors.

Hamel U; Kielwein G; Teschemacher H

1985-02-01

250

Demonstration of GABA immunoreactive cells in the inferior olive of baboons (Papio papio and Papio anubis).  

Science.gov (United States)

The distribution of gamma-aminobutyric acid (GABA)-like immunoreactivity was studied in semithin sections through the inferior olivary complex in two baboon species. About 5% of the olivary neurons were GABA-immunoreactive. The GABA-immunoreactive neurons differed from the large majority of olivary neurons by their smaller size and their lower contents of aspartate, as judged by analysis of alternate sections labelled with an aspartate antiserum. The present observations raise the possibility that in primates the GABAergic modulation of the activity of the climbing fibre system is effected not only by the previously described input from the cerebellar nuclei and other extrinsic sources, but that intrinsic neurons also participate. PMID:2671810

Walberg, F; Ottersen, O P

1989-06-19

251

Congenital abnormalities and multiple sclerosis  

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Background: There is a strong maternal parent-of-origin effect in determining susceptibility to multiple sclerosis (MS). One hypothesis is that an abnormal intrauterine milieu leading to impaired fetal development could plausibly also result in increased susceptibility to MS. A possible marker for t...

Ramagopalan, Sreeram V.; Guimond, Colleen; Criscuoli, Maria; Dyment, David A.

252

Chromosomal abnormalities associated with omphalocele.  

Science.gov (United States)

Fetuses with omphalocele have an increased risk for chromosomal abnormalities. The risk varies with maternal age, gestational age at diagnosis, association with umbilical cord cysts, complexity of associated anomalies, and the contents of omphalocele. There is considerable evidence that genetics contributes to the etiology of omphalocele. This article provides an overview of chromosomal abnormalities associated with omphalocele and a comprehensive review of associated full aneuploidy such as trisomy 18, trisomy 13, triploidy, trisomy 21, 45,X, 47,XXY, and 47,XXX, partial aneuploidy such as dup (3q), dup (11p), inv (11), dup (1q), del (1q), dup (4q), dup (5p), dup (6q), del (9p), dup (15q), dup(17q), Pallister-Killian syndrome with mosaic tetrasomy 12p and Miller-Dieker lissencephaly syndrome with deletion of 17p13.3, and uniparental disomy (UPD) such as UPD 11 and UPD 14. Omphalocele is a prominent marker for chromosomal abnormalities. Perinatal identification of omphalocele should alert chromosomal abnormalities and familial unbalanced translocations, and prompt thorough cytogenetic investigations and genetic counseling. PMID:17389182

Chen, Chih-Ping

2007-03-01

253

Ciliary abnormalities in respiratory disease.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

One hundred and sixty seven children, ranging in age from 5 weeks to 16 years, with chronic upper or lower respiratory tract problems, or both, were investigated for ciliary dyskinesia. Abnormal ciliary function was found in 18 cases all of whom had chronic lower respiratory disease and most of whom...

Buchdahl, R M; Reiser, J; Ingram, D; Rutman, A; Cole, P J; Warner, J O

254

Pap Test Results (Abnormal) -- Cancer  

Medline Plus

Full Text Available English - Abnormal Pap Test Results - Cancer 1 min 35 sec To Listen to the Audio or Read/Print/Save the Handout, Click on a Picture Below To ... A Health Guide for Women” by the National Cancer Institute

255

Oxytocin-immunoreactive innervation of identified neurons in the rat dorsal vagal complex.  

UK PubMed Central (United Kingdom)

BACKGROUND: Oxytocin (OXT) has been implicated in reproduction and social interactions and in the control of digestion and blood pressure. OXT-immunoreactive axons occur in the dorsal vagal complex (DVC; nucleus tractus solitarius, NTS, dorsal motor nucleus of the vagus, DMV, and area postrema, AP), which contains neurons that regulate autonomic homeostasis. The aim of the present work is to provide a systematic investigation of the OXT-immunoreactive innervation of dorsal motor nucleus of the vagus (DMV) neurons involved in the control of gastrointestinal (GI) function. METHODS: We studied DMV neurons identified by (i) prior injection of retrograde tracers in the stomach, ileum, or cervical vagus or (ii) induction of c-fos expression by glucoprivation with 2-deoxyglucose. Another subgroup of DMV neurons was identified electrophysiologically by stimulation of the cervical vagus and then juxtacellularly labeled with biotinamide. We used two- or three-color immunoperoxidase labeling for studies at the light microscopic level. KEY RESULTS: Close appositions from OXT-immunoreactive varicosities were found on the cell bodies, dendrites, and axons of DMV neurons that projected to the GI tract and that responded to 2-deoxyglucose and juxtacellularly labeled DMV neurons. Double staining for OXT and choline acetyltransferase revealed that OXT innervation was heavier in the caudal and lateral DMV than in other regions. OXT-immunoreactive varicosities also closely apposed a small subset of tyrosine hydroxylase-immunoreactive NTS and DMV neurons. CONCLUSIONS & INFERENCES: Our results provide the first anatomical evidence for direct OXT-immunoreactive innervation of GI-related neurons in the DMV.

Llewellyn-Smith IJ; Kellett DO; Jordan D; Browning KN; Travagli RA

2012-03-01

256

Distribution of high affinity choline transporter immunoreactivity in the primate central nervous system.  

UK PubMed Central (United Kingdom)

A mouse monoclonal antibody (clone 62-2E8) raised against a human recombinant high-affinity choline transporter (CHT)-glutathione-S-transferase fusion protein was used to determine the distribution of immunoreactive profiles containing this protein in the monkey central nervous system (CNS). Within the monkey telencephalon, CHT-immunoreactive perikarya were found in the striatum, nucleus accumbens, medial septum, vertical and horizontal limb nuclei of the diagonal band, nucleus basalis complex, and the bed nucleus of the stria terminalis. Dense fiber staining was observed within the islands of Calleja, olfactory tubercle, hippocampal complex, amygdala; moderate to light fiber staining was seen in iso- and limbic cortices. CHT-containing fibers were also present in sensory and limbic thalamic nuclei, preoptic and hypothalamic areas, and the floccular lobe of the cerebellum. In the brainstem, CHT-immunoreactive profiles were observed in the pedunculopontine and dorsolateral tegmental nuclei, the Edinger-Westphal, oculomotor, trochlear, trigeminal, abducens, facial, ambiguus, dorsal vagal motor, and hypoglossal nuclei. In the spinal cord, CHT-immunoreactive ventral horn motoneurons were seen in close apposition to intensely immunoreactive C-terminals at the level of the cervical spinal cord. CHT immunostaining revealed a similar distribution of labeled profiles in the aged human brain and spinal cord. Dual fluorescent confocal microscopy revealed that the majority of CHT immunoreactive neurons contained the specific cholinergic marker, choline acetyltransferase, at all levels of the monkey CNS. The present observations indicate that the present CHT antibody labels cholinergic structures within the primate CNS and provides an additional marker for the investigation of cholinergic neuronal function in aging and disease.

Kus L; Borys E; Ping Chu Y; Ferguson SM; Blakely RD; Emborg ME; Kordower JH; Levey AI; Mufson EJ

2003-08-01

257

Distribution of high affinity choline transporter immunoreactivity in the primate central nervous system.  

Science.gov (United States)

A mouse monoclonal antibody (clone 62-2E8) raised against a human recombinant high-affinity choline transporter (CHT)-glutathione-S-transferase fusion protein was used to determine the distribution of immunoreactive profiles containing this protein in the monkey central nervous system (CNS). Within the monkey telencephalon, CHT-immunoreactive perikarya were found in the striatum, nucleus accumbens, medial septum, vertical and horizontal limb nuclei of the diagonal band, nucleus basalis complex, and the bed nucleus of the stria terminalis. Dense fiber staining was observed within the islands of Calleja, olfactory tubercle, hippocampal complex, amygdala; moderate to light fiber staining was seen in iso- and limbic cortices. CHT-containing fibers were also present in sensory and limbic thalamic nuclei, preoptic and hypothalamic areas, and the floccular lobe of the cerebellum. In the brainstem, CHT-immunoreactive profiles were observed in the pedunculopontine and dorsolateral tegmental nuclei, the Edinger-Westphal, oculomotor, trochlear, trigeminal, abducens, facial, ambiguus, dorsal vagal motor, and hypoglossal nuclei. In the spinal cord, CHT-immunoreactive ventral horn motoneurons were seen in close apposition to intensely immunoreactive C-terminals at the level of the cervical spinal cord. CHT immunostaining revealed a similar distribution of labeled profiles in the aged human brain and spinal cord. Dual fluorescent confocal microscopy revealed that the majority of CHT immunoreactive neurons contained the specific cholinergic marker, choline acetyltransferase, at all levels of the monkey CNS. The present observations indicate that the present CHT antibody labels cholinergic structures within the primate CNS and provides an additional marker for the investigation of cholinergic neuronal function in aging and disease. PMID:12820166

Kus, Laura; Borys, Ewa; Ping Chu, Ya; Ferguson, Shawn M; Blakely, Randy D; Emborg, Marina E; Kordower, Jeffrey H; Levey, Allan I; Mufson, Elliott J

2003-08-25

258

Comparative anatomy of serotonin-like immunoreactive neurons in isopods: putative homologues in several species.  

Science.gov (United States)

It is now commonly accepted that the arthropod nervous system has evolved only once, and so homologies between crustacean and insect nervous systems can be meaningfully sought. To do this, we have examined the distribution of serotonin (5-hydroxytryptamine)-like immunoreactive neurons in the central nervous system (CNS) of four common British isopods. Two species of terrestrial woodlouse, Oniscus asellus and Armadillidium vulgare, the littoral sea slater, Ligia oceanica, and the aquatic water hoglouse, Asellus meridianus, all possess approximately 40 pairs of serotonin-like immunoreactive neurons, distributed throughout the CNS in a very similar pattern. Interspecific homology is clearly suggested. Serotonin-like immunoreactive neurons in the first (T1) and fourth (T4) thoracic ganglia are particularly prominent in each of the four species studied. Whole-mount immunohistochemistry shows that the pair of T1 neurons have large dorsolateral cell bodies and prominent neurites that project medially and then anteriorly, whereas the pair of T4 neurons have ventrolateral cell bodies and neurites that bifurcate to form a thin axon projecting anteriorly to terminate in T3 and a thick medial axon that projects posteriorly into the abdominal neuromeres of the terminal ganglion. Intracellular cobalt staining of these neurons reveals more of their arborizations: the T1 neurons send three processes anteriorly, which arborize in the brain and exist from the CNS via peripheral nerves, whereas the T4 neurons contribute considerably to the extensive pattern of serotonin-like immunoreactive fibres in T3-T6 ganglia. The overall pattern of serotonin-like immunoreactive neurons in the isopods is similar to that in decapod crustacea, and a number of putative homologies can be assigned. It is more difficult to homologize the isopod serotonin-like immunoreactive neurons with those in the insect CNS, but some stained brain and thoracic neurons share common cell body positions and axon trajectories in isopods, decapods, and insects and may therefore be homologous. PMID:7814675

Thompson, K S; Zeidler, M P; Bacon, J P

1994-09-22

259

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

Directory of Open Access Journals (Sweden)

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; Nikoli? Valentina; ?uleji? Vuk; Opri? Dejan; Rasuli? Lukas; Puškaš Laslo

2008-01-01

260

CT diagnosis of urachal abnormalities  

International Nuclear Information System (INIS)

Objective: To analyze CT manifestations of urachal abnormalities and to evaluate its clinical diagnostic value. Methods: CT findings of 23 cases of urachal abnormalities proven by surgery and pathology were retrospectively reviewed, the emphasis was focused on its location, shape, size, enhancement-pattern and radiological features in Retzius space. Results: All urachal abnormalities were located in the median line of Retzius space alone the course of the urachus (n=23). There was no positive CT finding in 1 case of patent urachus (n=1). Simple urachal cyst appeared as a homogeneous fluid-filled cavity with thin wall and no enhancement after the intravenous administration of contrast agency (n=2). Five cases of infected urachal cyst manifested as cysts with thickening and enhanced wall (n=5), 3 cases as multi-cyst lesion (n=3) and 2 cases as homogeneous mass (n=2). Some patch and strip appeared in Retzius space surrounding the lesions in these cases (n=10). The urachus neoplasms in 7 cases manifested as irregular soft-tissue masses in the midline of the apex of the bladder,with marked enhancement in 4 cases (n=4), mild enhancement in 2 cases (n=2) and no enhancement in 1 case (n=1). Three cases of urachal diverticulum accompanied with calculus appeared as intramural high-density lesions protruding into the bladder and the Retzius space simultaneously, 1 case with infected urachal cyst below umbilicus. Conclusion: Due to its special anatomic position, urachal abnormalities could be easily identified on CT preoperatively, but urachal cyst might mimic urachal tumor when infected sometimes. Familiarity with the anatomical basis and the CT features of urachal abnormalities would facilitate to establish the preoperative diagnosis correctly. (authors)

2005-01-01

 
 
 
 
261

Pulmonary enteric adenocarcinoma with villin brush border immunoreactivity: a case report and literature review.  

UK PubMed Central (United Kingdom)

Pulmonary enteric adenocarcinoma, a rare histologic type of primary pulmonary adenocarcinoma with only 16 cases reported to date, has to be differentiated from metastatic colorectal carcinoma. Here we report a case of pulmonary enteric adenocarcinoma which shows villin immunoreactivity in the brush border of tumor cells. As a marker for gastrointestinal adenocarcinoma, villin has been rarely found positive like this pattern in pulmonary adenocarcinomas. This case suggests brush border immunoreactivity of villin is possible in some cases of pulmonary enteric adenocarcinomas. We suggest pathological practitioners pay attention to it.

Lin D; Zhao Y; Li H; Xing X

2013-02-01

262

Substance P-like immunoreactivity in the nervous system of hydra  

DEFF Research Database (Denmark)

Using immunocytochemistry we find substance P-like material in nerve cells of hydra. These nerve cells are situated in the ectoderm of the basal disk and tentacles. Radioimmunoassay of hydra extracts gives dilution curves parallel to that of synthetic substance P, from which it can be calculated that one animal contains at least 0.6 fmol substance P-like immunoreactivity. After chromatography on Biogel P-100, the substance P-like immunoreactivity elutes as a peak in the void volume and a peak at the position of synthetic substance P.

Grimmelikhuijzen, C J; Balfe, A

1981-01-01

263

Immunoreactivity for high-affinity choline transporter colocalises with VAChT in human enteric nervous system.  

UK PubMed Central (United Kingdom)

Cholinergic nerves are identified by labelling molecules in the ACh synthesis, release and destruction pathway. Recently, antibodies against another molecule in this pathway have been developed. Choline reuptake at the synapse occurs via the high-affinity choline transporter (CHT1). CHT1 immunoreactivity is present in cholinergic nerve fibres containing vesicular acetylcholine transporter (VAChT) in the human and rat central nervous system and rat enteric nervous system. We have examined whether CHT1 immunoreactivity is present in nerve fibres in human intestine and whether it is colocalised with markers of cholinergic, tachykinergic or nitrergic circuitry. Human ileum and colon were fixed, sectioned and processed for fluorescence immunohistochemistry with antibodies against CHT1, class III beta-tubulin (TUJ1), synaptophysin, common choline acetyl-transferase (cChAT), VAChT, nitric oxide synthase (NOS), substance P (SP) and vasoactive intestinal peptide (VIP). CHT1 immunoreactivity was present in many nerve fibres in the circular and longitudinal muscle, myenteric and submucosal ganglia, submucosa and mucosa in human colon and ileum and colocalised with immunoreactivity for TUJ1 and synaptophysin confirming its presence in nerve fibres. In nerve fibres in myenteric ganglia and muscle, CHT1 immunoreactivity colocalised with immunoreactivity for VAChT and cChAT. Some colocalisation occurred with SP immunoreactivity, but little with immunoreactivity for VIP or NOS. In the mucosa, CHT1 immunoreactivity colocalised with that for VIP and SP in nerve fibres and was also present in vascular nerve fibres in the submucosa and on epithelial cells on the luminal border of crypts. The colocalisation of CHT1 immunoreactivity with VAChT immunoreactivity in cholinergic enteric nerves in the human bowel thus suggests that CHT1 represents another marker of cholinergic nerves.

Harrington AM; Lee M; Ong SY; Yong E; Farmer P; Peck CJ; Chow CW; Hutson JM; Southwell BR

2010-07-01

264

Endocrine abnormalities in anorexia nervosa.  

Science.gov (United States)

Anorexia nervosa (AN) is a psychiatric disease associated with notable medical complications and increased mortality. Endocrine abnormalities, including hypogonadotropic hypogonadism, hypercortisolemia, growth hormone resistance and sick euthyroid syndrome, mediate the clinical manifestations of this disease. Alterations in anorexigenic and orexigenic appetite-regulating pathways have also been described. Decreases in fat mass result in adipokine abnormalities. Although most of the endocrine changes that occur in AN represent physiologic adaptation to starvation, some persist after recovery and might contribute to susceptibility to AN recurrence. In this Review, we summarize key endocrine alterations in AN, with a particular focus on the profound bone loss that can occur in this disease. Although AN is increasingly prevalent among boys and men, the disorder predominantly affects girls and women who are, therefore, the focus of this Review. PMID:18542109

Lawson, Elizabeth A; Klibanski, Anne

2008-06-10

265

Endocrine abnormalities in anorexia nervosa.  

UK PubMed Central (United Kingdom)

Anorexia nervosa (AN) is a psychiatric disease associated with notable medical complications and increased mortality. Endocrine abnormalities, including hypogonadotropic hypogonadism, hypercortisolemia, growth hormone resistance and sick euthyroid syndrome, mediate the clinical manifestations of this disease. Alterations in anorexigenic and orexigenic appetite-regulating pathways have also been described. Decreases in fat mass result in adipokine abnormalities. Although most of the endocrine changes that occur in AN represent physiologic adaptation to starvation, some persist after recovery and might contribute to susceptibility to AN recurrence. In this Review, we summarize key endocrine alterations in AN, with a particular focus on the profound bone loss that can occur in this disease. Although AN is increasingly prevalent among boys and men, the disorder predominantly affects girls and women who are, therefore, the focus of this Review.

Lawson EA; Klibanski A

2008-07-01

266

Circuit for eliminating abnormal sound  

UK PubMed Central (United Kingdom)

A circuit for eliminating abnormal sound includes a video/audio processor, a D/A converter, a front-stage mute circuit, a changing delay device, a front-stage amplifier, a back-stage amplifier, a back-stage mute circuit, and a micro control unit. The circuit is mounted in a display device connected to a DVD player. When DVD player plays the video/audio signal having HDMI standard, the display device will generate an abnormal sound (pop voice or declining voice) during performing an operation such as pause, stop or play. Therefore, this circuit uses the changing delay device, the micro control unit, and the mute circuit to filter out the noise for providing a better outputting quality.

TSENG SHIH-HUA; YANG WEN-CHANG

267

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

268

Human serum dopamine beta-hydroxylase: correlation of enzymatic activity with immunoreactive protein in genetically defined samples.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

An antibody against human adrenal dopamine beta-hydroxylase (DBH) was used to quantitate immunoreactive DBH protein in human serum by an immunoprecipitation technique. A significant correlation was found between DBH enzyme activity and immunoreactive DBH protein in randomly selected serum samples (r...

Dunnette, J; Weinshilboum, R

269

Detection of FMRFamide-like immunoreactivities in the sea scallop Placopecten magellanicus by immunohistochemistry and western blot analysis.  

Science.gov (United States)

FMRFamide-like immunoreactivity was detected histochemically in the sea scallop Placopecten magellanicus. Most immunoreactivity was concentrated in the cerebral, pedal, and parietovisceral ganglia, particularly in the cortical cell bodies and in their fibers which extend into the central neuropile. Whole-mount immunofluorescence studies were used to localize concentrations of immunoreactive cells on the dorsal and ventral surfaces of each ganglion. Immunoreactivity was also detected in nerves emanating from the ganglia. Strong immunoreactivity was localized in peripheral organs, including the gut and gills of juvenile and adult scallops. Weak immunoreactivity was detected in the gonads, heart, and adductor muscle of the adults. A broad FMRFamide-like immunoreactive band of 2.5-8.2 kDa was detected by Western blotting of acetone extracts of the parietovisceral ganglia. In the presence of protease inhibitors, two FMRFamide-like immunoreactive bands (7.2-8.2 kDa and > 17 kDa) were obtained. Neither of these bands comigrated with the FMRFamide standard. It is concluded that peptides of the FMRFamide family are probably regulators of numerous central and peripheral functions in P. magellanicus. PMID:7648623

Too, C K; Croll, R P

1995-08-01

270

Detection of FMRFamide-like immunoreactivities in the sea scallop Placopecten magellanicus by immunohistochemistry and western blot analysis.  

UK PubMed Central (United Kingdom)

FMRFamide-like immunoreactivity was detected histochemically in the sea scallop Placopecten magellanicus. Most immunoreactivity was concentrated in the cerebral, pedal, and parietovisceral ganglia, particularly in the cortical cell bodies and in their fibers which extend into the central neuropile. Whole-mount immunofluorescence studies were used to localize concentrations of immunoreactive cells on the dorsal and ventral surfaces of each ganglion. Immunoreactivity was also detected in nerves emanating from the ganglia. Strong immunoreactivity was localized in peripheral organs, including the gut and gills of juvenile and adult scallops. Weak immunoreactivity was detected in the gonads, heart, and adductor muscle of the adults. A broad FMRFamide-like immunoreactive band of 2.5-8.2 kDa was detected by Western blotting of acetone extracts of the parietovisceral ganglia. In the presence of protease inhibitors, two FMRFamide-like immunoreactive bands (7.2-8.2 kDa and > 17 kDa) were obtained. Neither of these bands comigrated with the FMRFamide standard. It is concluded that peptides of the FMRFamide family are probably regulators of numerous central and peripheral functions in P. magellanicus.

Too CK; Croll RP

1995-08-01

271

Characteristics of abnormal glow discharges  

Energy Technology Data Exchange (ETDEWEB)

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

272

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

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...

Kolb, Helga; Zhang, Li; Dekorver, Laura; Cuenca Navarro, Nicolás

273

Distribution of Neuropeptide F-Like Immunoreactivity in the Eastern Subterranean Termite, Reticulitermes flavipes  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The nervous system and gut of worker, soldier and alate castes of the eastern subterranean termite, Reticulitermes flavipes Kollar (Isoptera: Rhinotermitidae) were examined for immunoreactivity to an antiserum to Helicoverpa zea (Boddie) (Leipidoptera: Noctuidae) MP-I (QAARPRF-NH2), a truncated form...

Nuss, Andrew B.; Forschler, Brian T.; Crim, Joe W.; Brown, Mark R.

274

The distribution and origin of substance P immunoreactive nerve fibres in the rat conjunctiva.  

UK PubMed Central (United Kingdom)

Indirect immunohistochemistry was used to examine the presence and origin of substance P immunoreactive nerve fibres in the rat conjunctiva. Fluorescent substance P immunoreactive nerve fibres were visualized both in the epithelium and stroma, their number being higher in the stroma where they were associated with blood vessels, the smooth muscle of Müller and the Meibomian glands. Ten to twenty percent of the ganglion cells in the trigeminal ganglion were immunoreactive to substance P, most of them being small in size. Sensory denervation by electrocoagulating the ophthalmic and maxillary branches of the trigeminal nerve caused a complete disappearance of the epithelial fibres and greatly reduced the number of the stromal fibres. These results indicate that the majority of the demonstrated fibres are sensory nerves originating from the trigeminal ganglion. Since sympathectomy had no detectable effect on the number or distribution of substance P immunoreactive nerve fibres, and since some of the fibres remained after sensory denervation it is suggested that at least some of the remaining fibres could be of parasympathetic origin.

Luhtala J; Uusitalo H

1991-11-01

275

The distribution and origin of substance P immunoreactive nerve fibres in the rat conjunctiva.  

Science.gov (United States)

Indirect immunohistochemistry was used to examine the presence and origin of substance P immunoreactive nerve fibres in the rat conjunctiva. Fluorescent substance P immunoreactive nerve fibres were visualized both in the epithelium and stroma, their number being higher in the stroma where they were associated with blood vessels, the smooth muscle of Müller and the Meibomian glands. Ten to twenty percent of the ganglion cells in the trigeminal ganglion were immunoreactive to substance P, most of them being small in size. Sensory denervation by electrocoagulating the ophthalmic and maxillary branches of the trigeminal nerve caused a complete disappearance of the epithelial fibres and greatly reduced the number of the stromal fibres. These results indicate that the majority of the demonstrated fibres are sensory nerves originating from the trigeminal ganglion. Since sympathectomy had no detectable effect on the number or distribution of substance P immunoreactive nerve fibres, and since some of the fibres remained after sensory denervation it is suggested that at least some of the remaining fibres could be of parasympathetic origin. PMID:1720739

Luhtala, J; Uusitalo, H

1991-11-01

276

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

Energy Technology Data Exchange (ETDEWEB)

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

277

Production of immunoreactive pancreatic growth hormone-releasing factor in small cell carcinoma of the lung.  

UK PubMed Central (United Kingdom)

A specific and sensitive radioimmunoassay for human pancreatic growth hormone-releasing factor (hpGRF) was developed. Using this radioimmunoassay, it was found that immunoreactive hpGRF (which is present in hypothalamic tissues) of at least two different molecular sizes is often produced by small cell carcinoma of the lung.

Yamaguchi K; Abe K; Suzuki M; Adachi I; Kimura S; Shimada A; Ohno H; Kanai A; Kameya T

1983-12-01

278

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

UK PubMed Central (United Kingdom)

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.

Roudenok V; Kühnel W; Rogov Y; Nerovnja A

1999-12-01

279

Distribution of oxytocin-immunoreactive neuronal elements in the rat spinal cord.  

UK PubMed Central (United Kingdom)

We investigated the distribution of oxytocin in rat spinal cord using immunocytochemistry and radioimmunoassay (RIA). Each segment of the spinal cord from cervical to coccygeal contained oxytocin-immunoreactive fibers. The Rexed laminae I and II of the dorsal horn showed moderate to intense immunoreactivity. A dense network was found around the central canal where some fibers apposed the ependyma. The autonomic centers of the spinal cord at the thoracolumbar and sacral segments were heavily innervated. Few fibers were found around the motoneurons. In the white matter, the immunoreactivity was localized mainly in the dorsal part of the lateral funiculus, in the pars funicularis of the nucleus intermediolateralis and in a longitudinal network of the lateral funiculus below the spinal cord surface. Some fibers from this network entered the pia mater. RIA measurements revealed that the cervical spinal cord had lower oxytocin content than that found in either the thoracic, lumbar, sacral or coccygeal region. Our results show that the distribution of oxytocin-immunoreactive fibers in the spinal cord correlates with anatomic locations related to nociceptive, autonomic and motor functions. We assume that oxytocin-containing axons play a role in secreting oxytocin directly into the liquor space of the spinal cord.

Jójárt J; Jójárt I; Boda K; Gálfi M; Mihály A; B-Baldauf Z; Vecsernyés M

2009-12-01

280

Neonatal screening strategy for cystic fibrosis using immunoreactive trypsinogen and direct gene analysis.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

OBJECTIVE--To assess the effectiveness of a two tier neonatal screening strategy for cystic fibrosis, which combines estimation of immunoreactive trypsinogen followed by direct gene analysis in dried blood spot samples collected at age 5 days. DESIGN--Prospective study of two tier screening strategy...

Ranieri, E; Ryall, R G; Morris, C P; Nelson, P V; Carey, W F; Pollard, A C; Robertson, E F

 
 
 
 
281

Endocan immunoreactivity in the mouse brain: Method for identifying nonfunctional blood vessels.  

UK PubMed Central (United Kingdom)

Endocan is a secreted proteoglycan that has been shown to indicate angiogenic activity: remodeling in several tumor types in humans and mice. Serum endocan levels also indicate prognosis and has been proposed as a biomarker for certain cancers. Recently, monoclonal antibodies directed against mouse endocan have been developed allowing for further characterization of endocan function and potentially as a marker for angiogenesis through immunoreactivity in endothelial tip cells. The results of the current study show that endocan immunoreactivity in the mouse brain is present in blood vascular networks including but not limited to the cortex, hippocampus and paraventricular nucleus of the hypothalamus in C57BL/6J and FVB/N mice. Endocan immunoreactivity did not vary during postnatal development or by sex. Interestingly, after vascular perfusion with fluorescein isothiocyanate (FITC), endothelial cells positive for FITC were immunonegative for endocan suggesting FITC interference with the immunohistochemistry. A small number of FITC-negative blood vessels were endocan immunoreactive suggesting the identification of new blood vessels that are not yet functional. The current study shows that endocan is normally present in the mouse brain and prior vascular perfusion with FITC may provide a useful tool for identify newly forming blood vessels.

Frahm KA; Nash CP; Tobet SA

2013-09-01

282

Peripheral plasma immunoreactive 6-oxo-prostaglandin F1 alpha and gynaecological tumours.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Peripheral plasma levels of immunoreactive 6-oxo-PGF1 alpha, the stable hydrolysis product of prostacyclin, were significantly higher in female patients with tumours of the genital tract than in normal controls. In the groups with malignant tumours, these high levels declined after operation and/or ...

Alam, M.; Jogee, M.; MacGregor, W. G.; Dowdell, J. W.; Elder, M. G.; Myatt, L.

283

Implications of lipid moiety in oligomerization and immunoreactivities of GPI-anchored proteins.  

UK PubMed Central (United Kingdom)

Glycosylphosphatidylinositol (GPI) enriches GPI-anchored proteins (GPI-AP) in lipid rafts by intimate interaction of its lipid moiety with sphingolipids and cholesterol. In addition to such lipid-lipid interactions, it has been reported that GPI may interact with protein moiety linked to GPI and affect protein conformations because GPI delipidation reduced immunoreactivities of protein. Here, we report that GPI-APs that have not undergone fatty acid remodeling exhibit reduced immunoreactivities in Western blotting, similar to delipidated proteins, compared with normal remodeled GPI-APs. In contrast, immunostaining in flow cytometry and immunoprecipitation did not show significant differences between remodeled and unremodeled GPI-APs. Moreover, detection with premixed primary/secondary antibody complexes or Fab fragments eliminated this difference in Western blotting. These results indicate that normally remodeled GPI enhanced oligomerization of GPI-APs and that inefficient oligomerization of unremodeled GPI-APs was responsible for reduced immunoreactivities. Moreover, the reduction in immunoreactivities of delipidated GPI-APs was most likely caused by the same effect. Finally, by chemical cross-linking of surface proteins in living cells and cell killing assay using a pore-forming bacterial toxin, we showed that enhanced oligomerization by GPI-remodeling occurs under a physiological membrane environment. Thus, this study clarifies the significance of GPI fatty acid remodeling in oligomerization of GPI-APs and provides useful information for technical studies of these cell components.

Seong J; Wang Y; Kinoshita T; Maeda Y

2013-04-01

284

Nitric oxide synthase-immunoreactive neurons in human and porcine respiratory tract  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The presence of nitric oxide synthase (NO-synthase), the enzyme responsible for the production of nitric oxide (NO) from L-arginine, is shown immunocytochemically in the intrinsic neurons of the human and porcine respiratory tract. NO-synthase immunoreactivity is demonstrated in a subpopulation of n...

285

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

UK PubMed Central (United Kingdom)

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.

2012-07-01

286

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

International Nuclear Information System (INIS)

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

1985-09-25

287

Neuropeptide Y-immunoreactive neurons in the retina of two Australian lizards.  

Science.gov (United States)

Wholemounts and sectioned retina from adults of two lizard species, Pogona vitticeps and Varanus gouldii, were studied by immunohistochemistry for neuropeptide Y (NPY)-like immunoreactivity. In both species the morphology of two classes of amacrine cells (types A and B) were described. Cell somata were located mostly in the inner nuclear layer (INL) but were occasionally displaced into the ganglion cell layer (GCL). In the Pogona retina, type A cells had large somata and dendritic arbor that branched in sublamina (S) 1 and 2/3 of the inner plexiform layer (IPL). Type B amacrine cells had smaller somata and dendritic arbor branching mostly in S5 of the IPL. In the Varanus retina, the levels of dendritic branching of types A and B amacrine cells in the IPL were similar to those in Pogona although branching in the middle of the IPL occurred at S3. NPY-immunoreactive cells with small somata and narrow to medium sized dendritic fields were predominant. Unclassified cells also displayed NPY-like immunoreactivity; however, their dendritic morphology could not be determined due to the faint and inconsistent staining. In transverse retinal sections three bands of NPY-like immunoreactivity were evident in the IPL of both species, to which the unclassified cells also contributed. In both species type A cells were most numerous. Total NPY-immunoreactive cells were estimated to be 8,600 in Pogona and 32,860 in Varanus. In both species types A and B cells were non-uniformly distributed across the retina. The most apparent non-uniformity in distribution was observed in type A cells in Varanus. Peak cell density was found across the horizontal meridian of the retina from where cell density decreased towards the dorsal and ventral retinal margins. The results of this study provide evidence for the presence of NPY-immunoreactive amacrine cells in the lizard retina of which two types were morphologically characterized. Cross-species comparisons were also made among NPY-immunoreactive amacrine cells, and their possible function/s discussed. PMID:8068405

Straznicky, C; Hiscock, J

1994-05-01

288

Neuropeptide Y-immunoreactive neurons in the retina of two Australian lizards.  

UK PubMed Central (United Kingdom)

Wholemounts and sectioned retina from adults of two lizard species, Pogona vitticeps and Varanus gouldii, were studied by immunohistochemistry for neuropeptide Y (NPY)-like immunoreactivity. In both species the morphology of two classes of amacrine cells (types A and B) were described. Cell somata were located mostly in the inner nuclear layer (INL) but were occasionally displaced into the ganglion cell layer (GCL). In the Pogona retina, type A cells had large somata and dendritic arbor that branched in sublamina (S) 1 and 2/3 of the inner plexiform layer (IPL). Type B amacrine cells had smaller somata and dendritic arbor branching mostly in S5 of the IPL. In the Varanus retina, the levels of dendritic branching of types A and B amacrine cells in the IPL were similar to those in Pogona although branching in the middle of the IPL occurred at S3. NPY-immunoreactive cells with small somata and narrow to medium sized dendritic fields were predominant. Unclassified cells also displayed NPY-like immunoreactivity; however, their dendritic morphology could not be determined due to the faint and inconsistent staining. In transverse retinal sections three bands of NPY-like immunoreactivity were evident in the IPL of both species, to which the unclassified cells also contributed. In both species type A cells were most numerous. Total NPY-immunoreactive cells were estimated to be 8,600 in Pogona and 32,860 in Varanus. In both species types A and B cells were non-uniformly distributed across the retina. The most apparent non-uniformity in distribution was observed in type A cells in Varanus. Peak cell density was found across the horizontal meridian of the retina from where cell density decreased towards the dorsal and ventral retinal margins. The results of this study provide evidence for the presence of NPY-immunoreactive amacrine cells in the lizard retina of which two types were morphologically characterized. Cross-species comparisons were also made among NPY-immunoreactive amacrine cells, and their possible function/s discussed.

Straznicky C; Hiscock J

1994-05-01

289

Substance P-immunoreactive neurons in the retina of two lizards.  

UK PubMed Central (United Kingdom)

The dendritic morphology and retinal distribution of substance P(SP)-immunoreactive neurons was determined in two Australian lizard species Pogona vitticeps and Varanus gouldii, by using immunohistochemistry on retinal wholemounts and sectioned materials. In both species, two classes of SP-immunoreactive neurons were described in the inner nuclear layer (INL) and classified as amacrine cells (types A and B). Type A amacrine cells had large somata and wide-field, bistratified dendrites branching in sublaminas 1 and 5 of the inner plexiform layer (IPL). Their morphology and retinal distribution differed between the two species. Type B amacrine cells in both species had small somata and small-field dendritic branching. A population of SP-immunoreactive neurons with classical ganglion cell morphology were identified in the ganglion cell layer (GCL). Immunostained ganglion cells occurred in larger numbers of Varanus gouldii than in Pogona vitticeps. In both species type B SP cells were the most numerous and were estimated to be about 60,000-70,000. They were distributed non-uniformly with a high density band across the horizontal meridian of the retina, from where the density decreased towards the dorsal and ventral retinal margins. In both species type A amacrine cells occurred in small numbers distributed sparsely in the peripheral retina. The faint immunostaining of SP-immunoreactive neurons in the GCL, did not allow us to reliably determine their numbers and retinal distribution. The functional significance of SP-immunoreactive amacrine and ganglion cells in the lizard retina remains to be determined.

Hiscock J; Straznicky C

1991-07-01

290

Substance P-immunoreactive neurons in the retina of two lizards.  

Science.gov (United States)

The dendritic morphology and retinal distribution of substance P(SP)-immunoreactive neurons was determined in two Australian lizard species Pogona vitticeps and Varanus gouldii, by using immunohistochemistry on retinal wholemounts and sectioned materials. In both species, two classes of SP-immunoreactive neurons were described in the inner nuclear layer (INL) and classified as amacrine cells (types A and B). Type A amacrine cells had large somata and wide-field, bistratified dendrites branching in sublaminas 1 and 5 of the inner plexiform layer (IPL). Their morphology and retinal distribution differed between the two species. Type B amacrine cells in both species had small somata and small-field dendritic branching. A population of SP-immunoreactive neurons with classical ganglion cell morphology were identified in the ganglion cell layer (GCL). Immunostained ganglion cells occurred in larger numbers of Varanus gouldii than in Pogona vitticeps. In both species type B SP cells were the most numerous and were estimated to be about 60,000-70,000. They were distributed non-uniformly with a high density band across the horizontal meridian of the retina, from where the density decreased towards the dorsal and ventral retinal margins. In both species type A amacrine cells occurred in small numbers distributed sparsely in the peripheral retina. The faint immunostaining of SP-immunoreactive neurons in the GCL, did not allow us to reliably determine their numbers and retinal distribution. The functional significance of SP-immunoreactive amacrine and ganglion cells in the lizard retina remains to be determined. PMID:1720010

Hiscock, J; Straznicky, C

1991-07-01

291

Catecholaminergic system innervates galanin-immunoreactive neurons in the human diencephalon.  

Science.gov (United States)

Galanin released into the hypophysial portal circulation in the hypothalamus may function as a hypophysiotropic factor regulating the anterior pituitary function or it may function as a neurotransmitter/neuromodulator acting at synaptic sites regulating neuronal activity of many neurons in the brain. Catecholamines (adrenaline, noradrenaline, and dopamine) primarily regulate anterior pituitary functions indirectly via innervating hypophysiotropic neurons. The aim of the present studies was to explore with double-label immunocytochemistry if, as in rodents, catecholamines interact with galanin in the human diencephalon. Due to the long post-mortem period and subsequent lack of optimal preservation of the cell membranes in the brain, electron microscopy could not be employed to show the presence of catecholaminergic-immunoreactive synapses on galanin-immunoreactive neurons. Therefore, we used light microscopic immunocytochemistry and high-magnification microscopy with oil immersion to identify putative juxtapositions between catecholamines and galanin-utilizing antisera against key enzymes of catecholamine synthesis (tyrosine hydroxylase (TH), representing all three catecholamines; dopamine-beta-hydroxylase (DBH), representing noradrenaline; and phenylethanolamine-N-methyltransferase (PNMT), representing adrenaline) and galanin. Our studies show that among the three catecholamines, dopamine is the most abundant and the vast majority of catecholaminergic contacts on galanin-immunoreactive neurons is dopaminergic. The number of DBH-immunoreactive contacts is less and the number of PNMT-immunopositive contacts is negligible. Among the hypothalamic regions, the periventricular region above the infundibulum (infundibular or arcuate nucleus) contained the largest number of contacts. These en passant-type intimate associations between catecholamine- and galanin-immunoreactive neuronal elements may be functional synapses and may provide the morphological basis for the catecholamine-mediated galanin release. PMID:23415787

Merchenthaler, I; Rotoli, G; Peroski, M; Grignol, G; Dudas, B

2013-02-13

292

Increased total volume and dopamine ?-hydroxylase immunoreactivity of carotid body in spontaneously hypertensive rats.  

UK PubMed Central (United Kingdom)

Under hypertension, it has been reported that the carotid body (CB) is enlarged and noradrenaline (NA) content in CB is increased. Therefore, it is hypothesized that morphological and neurochemical changes in CB are induced in hypertensive animal models. In the present study, we examined the morphological features and dopamine ?-hydroxylase (DBH) immunoreactivity in CB of spontaneously hypertensive rats (SHR/Izm) and Wistar Kyoto rats (WKY/Izm). The CB of SHR/Izm was elongated in terms of the cross section of center and was enlarged in the reconstructed images compared with that of WKY/Izm, and the total volume of CB in SHR/Izm (0.048 ± 0.004 mmł) was significantly (p<0.05) increased compared with the value in WKY/Izm (0.032 ± 0.006 mmł). By immunohistochemistry, immunoreactivity for tyrosine hydroxylase in CB was mainly observed in glomus cells and the immunostaining properties were similar between WKY/Izm and SHR/Izm. On the other hand, DBH immunoreactivity was mainly observed in nerve fibers around blood vessels and observed in a few glomus cells in CB of WKY/Izm. The number of glomus cells with strong DBH immunoreactivity was increased in SHR/Izm compared with that in WKY/Izm. In conclusion, the present study exhibited the enlargement of CB as three-dimensional image and revealed the enhanced immunoreactivity for DBH of glomus cells in SHR/Izm. These results suggest that the morphology of CB is affected by the effect of sympathetic nerve and that the signal transduction from CB is regulated by NA in glomus cells under hypertensive conditions.

Kato K; Wakai J; Matsuda H; Kusakabe T; Yamamoto Y

2012-07-01

293

Analysis of metallothionein and vimentin immunoreactivity in pharyngeal squamous cell carcinoma and its microenvironment.  

UK PubMed Central (United Kingdom)

Metallothionein (MT) has been shown to have pro-proliferative anti-apoptotic activity and to be involved in microenvironment remodeling. The aim of this study has been to determine whether the changes in MT and vimentin immunoreactivity observed in cancer and its microenvironment are related to the local spread of the disease. The immunoreactivity levels of both MT and vimentin were evaluated together with CD56 and CD57 antigens in 49 tissue samples taken from patients with squamous cell carcinoma originating from the palatine tonsils and in 20 tissue samples derived from patients with chronic tonsillitis (the reference group). MT immunoreactivity levels were statistically significantly higher in the tissue samples from squamous cell carcinoma than in those of the reference group and also higher in the squamous cell carcinoma samples compared with the stromal samples. Moreover, stromal fibroblasts exhibited high vimentin and MT immunoreactivity levels. Statistically significantly higher MT immunoreactivity levels within the tumor cells were identified in patients with the presence of lymph node metastases in contrast to those patients without such metastases. Vimentin was detected in both the tumor and the stromal tissue samples and presented an interesting pattern of staining strongly expressed within the stroma and the septal architecture of the tumor. The number of CD56- and CD57-positive lymphocytes identified in tissue samples both from squamous cell carcinoma and from the stroma was statistically significantly lower than that in the reference group. MT expression by tumor cells is thus associated with an aggressive phenotype of the tumor and the ability to create metastases.

Dutsch-Wicherek M; Lazar A; Tomaszewska R; Kazmierczak W; Wicherek L

2013-05-01

294

Epidural interferon gamma-immunoreactivity: a biomarker for lumbar nerve root irritation.  

UK PubMed Central (United Kingdom)

STUDY DESIGN: Prospective observational cohort. OBJECTIVE: Correlate epidural inflammatory cytokines with the clinical response to epidural steroid injection in patients with lumbar nerve root irritation. SUMMARY OF BACKGROUND DATA: Some back pain syndromes are thought to be associated with activation of inflammatory pathways and others may be associated with primary mechanical derangements. Human studies providing detailed evidence for the primary inflammatory causation, which may be best treated with anti-inflammatory strategies, are lacking. There are currently no accurate diagnostic tests to predict the response to epidural steroid injection or surgical intervention in back pain and sciatica syndromes. METHODS.: Forty-seven consecutive patients with lumbar degenerative changes and low back and/or leg pain were prospectively enrolled. An epidural lavage was performed, followed by injection of marcaine/depo-medrol. Subjects scored their pain before and 3 months after the procedure. The immunoreactivity of an array of cytokines was measured in lavage samples and compared with clinical response to the therapeutic injection. Ten subjects underwent repeat epidural lavage sampling 3 months after the steroid injection. RESULTS: Interferon gamma (IFNgamma) was the most consistently detected cytokine. IFNgamma-immunoreactivity also highly correlated with reported reduction of pain 3-months after the epidural steroid injection. In subjects reporting significant pain relief (>50%) from the injection, mean [IFNgamma] was significantly greater compared with patients experiencing no significant relief. The IFNgamma-immunoreactivity in repeat lavage samples decreased to trace residual concentrations in patients who reported pain relief from the steroid injection. CONCLUSION: The presence of epidural IFNgamma-immunoreactivity corresponding to >10 pg/mL predicted significant pain relief after epidural steroid injection with >95% accuracy. These results suggest that IFNgamma may be part of a biochemical cascade triggering pain in sciatica; IFNgamma-immunoreactivity may aid as a biomarker for predicting the response to steroid therapy and/or surgical intervention, and may serve as a future therapeutic target.

Scuderi GJ; Cuellar JM; Cuellar VG; Yeomans DC; Carragee EJ; Angst MS

2009-10-01

295

Immunoreactivity for calretinin and keratins in desmoid fibromatosis and other myofibroblastic tumors: a diagnostic pitfall.  

UK PubMed Central (United Kingdom)

Calretinin is an intracellular calcium-binding EF-hand protein of the calmodulin superfamily. It plays a role in diverse cellular functions, including message targeting and intracellular calcium signaling. It is expressed in the mesothelium, mast cells, some neural cells, and fat cells, among others. Because of its relative specificity for mesothelial neoplasms, calretinin is widely used as one of the primary immunohistochemical markers for malignant mesothelioma and in differentiating it from adenocarcinoma. On the basis of our sporadic observation on calretinin immunoreactivity in desmoid fibromatosis, we systematically evaluated calretinin, keratin cocktail (AE1/AE3), and WT1 immunoreactivity in 268 fibroblastic/myofibroblastic neoplasms. Calretinin was observed in 75% (44/58) of desmoid fibromatosis, 50% (21/42) of proliferative fasciitis, 23% (8/35) of nodular fasciitis, 33% (13/40) of benign fibrous histiocytoma, 35% (22/62) of malignant fibrous histiocytoma, and 13% (4/31) of solitary fibrous tumors but not in normal connective tissue fibroblasts at various sites. Keratin AE1/AE3 immunoreactivity was also commonly (6/13) present in the large ganglion-like cells of proliferative fasciitis and sometimes in nodular fasciitis (3/35), solitary fibrous tumor (3/27), and malignant fibrous histiocytoma (9/62). Nuclear immunoreactivity for WT1 or keratin 5 positivity was not detected in myofibroblastic tumors. On the basis of these observations, it can be concluded that calretinin and focal keratin immunoreactivity is fairly common in benign and malignant fibroblastic and myofibroblastic lesions. Calretinin-positive and keratin-positive spindle cells in desmoid and nodular fasciitis or calretinin-positive ganglion-like cells in proliferative fasciitis should not be confused with elements of epithelioid or sarcomatoid mesothelioma. These diagnostic pitfalls can be avoided with careful observation of morphology, quantitative differences in keratin expression, and use of additional immunohistochemical markers such keratin 5 and WT1 to verify true epithelial and mesothelial differentiation typical of mesothelioma.

Barak S; Wang Z; Miettinen M

2012-09-01

296

Immunoreactivity for calretinin and keratins in desmoid fibromatosis and other myofibroblastic tumors: a diagnostic pitfall.  

Science.gov (United States)

Calretinin is an intracellular calcium-binding EF-hand protein of the calmodulin superfamily. It plays a role in diverse cellular functions, including message targeting and intracellular calcium signaling. It is expressed in the mesothelium, mast cells, some neural cells, and fat cells, among others. Because of its relative specificity for mesothelial neoplasms, calretinin is widely used as one of the primary immunohistochemical markers for malignant mesothelioma and in differentiating it from adenocarcinoma. On the basis of our sporadic observation on calretinin immunoreactivity in desmoid fibromatosis, we systematically evaluated calretinin, keratin cocktail (AE1/AE3), and WT1 immunoreactivity in 268 fibroblastic/myofibroblastic neoplasms. Calretinin was observed in 75% (44/58) of desmoid fibromatosis, 50% (21/42) of proliferative fasciitis, 23% (8/35) of nodular fasciitis, 33% (13/40) of benign fibrous histiocytoma, 35% (22/62) of malignant fibrous histiocytoma, and 13% (4/31) of solitary fibrous tumors but not in normal connective tissue fibroblasts at various sites. Keratin AE1/AE3 immunoreactivity was also commonly (6/13) present in the large ganglion-like cells of proliferative fasciitis and sometimes in nodular fasciitis (3/35), solitary fibrous tumor (3/27), and malignant fibrous histiocytoma (9/62). Nuclear immunoreactivity for WT1 or keratin 5 positivity was not detected in myofibroblastic tumors. On the basis of these observations, it can be concluded that calretinin and focal keratin immunoreactivity is fairly common in benign and malignant fibroblastic and myofibroblastic lesions. Calretinin-positive and keratin-positive spindle cells in desmoid and nodular fasciitis or calretinin-positive ganglion-like cells in proliferative fasciitis should not be confused with elements of epithelioid or sarcomatoid mesothelioma. These diagnostic pitfalls can be avoided with careful observation of morphology, quantitative differences in keratin expression, and use of additional immunohistochemical markers such keratin 5 and WT1 to verify true epithelial and mesothelial differentiation typical of mesothelioma. PMID:22531174

Barak, Stephanie; Wang, Zengfeng; Miettinen, Markku

2012-09-01

297

Analysis of metallothionein and vimentin immunoreactivity in pharyngeal squamous cell carcinoma and its microenvironment.  

Science.gov (United States)

Metallothionein (MT) has been shown to have pro-proliferative anti-apoptotic activity and to be involved in microenvironment remodeling. The aim of this study has been to determine whether the changes in MT and vimentin immunoreactivity observed in cancer and its microenvironment are related to the local spread of the disease. The immunoreactivity levels of both MT and vimentin were evaluated together with CD56 and CD57 antigens in 49 tissue samples taken from patients with squamous cell carcinoma originating from the palatine tonsils and in 20 tissue samples derived from patients with chronic tonsillitis (the reference group). MT immunoreactivity levels were statistically significantly higher in the tissue samples from squamous cell carcinoma than in those of the reference group and also higher in the squamous cell carcinoma samples compared with the stromal samples. Moreover, stromal fibroblasts exhibited high vimentin and MT immunoreactivity levels. Statistically significantly higher MT immunoreactivity levels within the tumor cells were identified in patients with the presence of lymph node metastases in contrast to those patients without such metastases. Vimentin was detected in both the tumor and the stromal tissue samples and presented an interesting pattern of staining strongly expressed within the stroma and the septal architecture of the tumor. The number of CD56- and CD57-positive lymphocytes identified in tissue samples both from squamous cell carcinoma and from the stroma was statistically significantly lower than that in the reference group. MT expression by tumor cells is thus associated with an aggressive phenotype of the tumor and the ability to create metastases. PMID:23397427

Dutsch-Wicherek, Magdalena; Lazar, Agata; Tomaszewska, Romana; Kazmierczak, Wojciech; Wicherek, Lukasz

2013-02-09

298

Carbamazepine for acute psychosis with eeg abnormalities  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Aim. To investigate the efficacy of carbamazepine as adjuvant drug therapy in acute paranoid psychosis with associated EEG abnormalities, compared to sole antipsychotic treatment. Methods. Eleven medication-naive patients diagnosed with acute paranoid psychosis with associated EEG abnormalities were...

Ivkovi? Maja; Damjanovi? Aleksandar; Marinkovi? Dragan; Paunovi? Vladimir R.

299

Cardiovascular abnormalities in Klinefelter Syndrome.  

UK PubMed Central (United Kingdom)

BACKGROUND: Several epidemiological studies have demonstrated an increased mortality from cardiovascular causes in patients with Klinefelter Syndrome (KS). Little information is available about the nature of the underlying cardiovascular abnormalities. Aim of the study was to investigate exercise performance, left ventricular architecture and function, vascular reactivity, and carotid intima-media thickness in a group of patients with KS. MATERIALS AND METHODS: Sixty-nine patients with KS and 48 age-matched controls participated in our population-controlled study. Forty-eight Klinefelter subjects were on testosterone treatment at the time of the investigation while 21 were naive and underwent a complete Doppler echocardiographic examination, a cardiopulmonary exercise test as well as a vascular study including measures of carotid intima-media thickness and endothelial function with flow-mediated dilation of the brachial artery. Patients with KS on testosterone therapy (n=48) were also matched against a population of men with treated secondary hypogonadism (n=21). RESULTS: Patients with KS exhibited a wide array of cardiovascular abnormalities including left ventricular diastolic dysfunction, reduced maximal oxygen consumption (p<0.01), increased intima-media thickness (p<0.05) (-34% and +42% vs. controls, respectively) and a high prevalence of chronotropic incompetence (55% of patients, p<0.01). No significant difference was found between treated and untreated KS in variance with men treated for secondary hypogonadism. CONCLUSION: Left ventricular diastolic dysfunction, impaired cardiopulmonary performance, chronotropic incompetence, and increased intima-media thickness suggest that cardiovascular abnormalities are a common finding in KS that is not reversed by testosterone replacement therapy and may represent the pathophysiological underpinnings of the increased risk of dying from heart disease.

Pasquali D; Arcopinto M; Renzullo A; Rotondi M; Accardo G; Salzano A; Esposito D; Saldamarco L; Isidori AM; Marra AM; Ruvolo A; Napoli R; Bossone E; Lenzi A; Baliga RR; Saccŕ L; Cittadini A

2013-09-01

300

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

 
 
 
 
301

Selective subcortical abnormalities in autism.  

UK PubMed Central (United Kingdom)

Nine physically healthy, adult autistic men, with normal or near normal intelligence, and 13 healthy male controls were examined in a CT brain scan study. CT scans were analysed with a fully automated computer-assisted program, and regional brain radiodensities were measured with careful attention to artefacts. Autistic patients revealed significantly larger third, but not lateral, ventricular size and significantly lower mean caudate, but equivalent mean frontal and thalamic, radiodensities compared to controls. The sizes of the Sylvian fissures and interhemispheric fissure were equivalent between groups. The findings are consistent with selective subcortical abnormalities in autism.

Jacobson R; Le Couteur A; Howlin P; Rutter M

1988-02-01

302

Abnormal axillary lymph nodes in rheumatoid arthritis  

International Nuclear Information System (INIS)

[en] Mammary radiography was performed in 56 patients with arthritis, predominantly rheumatoid arthritis. Definite abnormal axillary lymph nodes were found in 24 and probably abnormal nodes in 8. Abnormal nodes were characterized by increased attenuation, rounded shape and absence of fatty replacement. (Auth)

1980-01-01

303

Insulin resistance and abnormal ovarian responses to human chorionic gonadotropin in chronically anovulatory women.  

UK PubMed Central (United Kingdom)

We studied the interrelationships between insulin resistance, obesity, and abnormal ovarian androgen secretion in chronically anovulatory women with clinical or biochemical evidence of hyperandrogenism. Four groups of six subjects each were studied: (1) normal weight (within 10% ideal body weight) anovulatory, (2) obese (greater than 120% ideal body weight) anovulatory, (3) normal weight eumenorrheic, and (4) obese eumenorrheic. After dexamethasone suppression, human chorionic gonadotropin (2000 IU/1.5m2 body surface area intramuscularly) was administered to each subject. Serum testosterone levels were subsequently determined hourly for 17 hours. On a separate occasion, an oral glucose tolerance test was administered to five subjects from each group. Serum glucose and immunoreactive insulin levels were determined before and after the ingestion of a standard 100 gm glucose load. As a group, the anovulatory women had higher (p less than 0.05) basal testosterone levels (1005 +/- 97 pg/ml) than did the ovulatory women (241 +/- 21 pg/ml) (values +/- SE). Obesity per se was not associated with increased basal testosterone levels. Testosterone levels rose in response to human chorionic gonadotropin (p less than 0.005) only in obese anovulatory women, reached maximal levels after 3 hours, and subsequently remained stable. Basal immunoreactive insulin levels were elevated (p less than 0.05) only in obese anovulatory women (52.4 +/- 20 microU/ml) compared with obese eumenorrheic (8.7 +/- 1.0 microU/ml), normal weight anovulatory (5.8 +/- 2.4 microU/ml), and normal weight eumenorrheic (4.6 +/- 0.4 microU/ml) women. Similarly, maximal increases in immunoreactive insulin levels after glucose ingestion were significantly greater (p less than 0.01) in obese anovulatory women compared with other groups. Of note is the observation that maximal changes in testosterone observed within the first 3 hours after human chorionic gonadotropin and maximal changes in insulin were correlated (r = 0.91, p less than 0.01). These data suggest that (1) both insulin resistance and an abnormal acute response to human chorionic gonadotropin are seen only in obese anovulatory women and (2) the degree to which these two abnormalities are manifested is clearly correlated. The mechanism(s) responsible for this interrelationship, as well as the underlying cause(s) of these biochemical defects, remain to be elucidated.

Kustin J; Kazer RR; Hoffman DI; Chatterton RT Jr; Haan JN; Green OC; Rebar RW

1987-12-01

304

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

2013-06-01

305

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

UK PubMed Central (United Kingdom)

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.

Latorre R; Mazzoni M; De Giorgio R; Vallorani C; Bonaldo A; Gatta PP; Corinaldesi R; Ruggeri E; Bernardini C; Chiocchetti R; Sternini C; Clavenzani P

2013-06-01

306

The distribution of glutamic acid decarboxylase immunoreactivity in the diencephalon of the opossum and rabbit.  

UK PubMed Central (United Kingdom)

We have examined the distribution of neurons and terminals immunoreactive for glutamic acid decarboxylase (GAD) in the thalamus and adjacent structures of the opossum (Didelphis virginiana) and the rabbit and have compared this distribution with the distributions we described previously for the cat and bushbaby (Galago senegalensis). The significance of these experiments depends, first, on the fact that GAD is the synthetic enzyme for GABA, and therefore that GAD immunoreactivity is a marker for GABAergic inhibitory neurons, and second, on previous findings that suggest that GABAergic neurons in the dorsal thalamus are local circuit neurons. In both cat and Galago, GAD-immunoreactive neurons are distributed essentially throughout the entire thalamus. In the opossum, GAD neurons are chiefly confined to the dorsal lateral geniculate nucleus and the lateral extremity of the lateral posterior nucleus. The distribution of GAD neurons in the rabbit is intermediate between that found in the opossum on the one hand and cat and Galago on the other. Like opossum, about 25% of the neurons in the lateral geniculate nucleus of rabbit are GAD immunoreactive. Unlike opossum, however, as many as 18% of the cells in the ventral posterior nucleus of the rabbit are GAD immunoreactive, and scattered cells are also labeled in other thalamic areas, such as the medial geniculate and the lateral group. Aside from the findings in the dorsal thalamus, the chief observation is that GAD-immunoreactive neurons and/or terminals densely fill all principal targets of the optic tract, including the ventral lateral geniculate nucleus; the superficial gray layer of the superior colliculus; the anterior, posterior, and olivary pretectal nuclei; the nucleus of the optic tract; and the medial and lateral terminal nuclei of the accessory optic tract. These results support the idea first put forward by Cajal that local circuit neurons increase in number during the course of the evolution of complex mammalian brains. If we can assume that the conservative opossum retains characteristics reflecting an early stage of mammalian evolution, the results suggest that thalamic local circuit neurons arose first in the visual system and only later in evolution spread throughout the thalamus.

Penny GR; Conley M; Schmechel DE; Diamond IT

1984-09-01

307

The distribution of glutamic acid decarboxylase immunoreactivity in the diencephalon of the opossum and rabbit.  

Science.gov (United States)

We have examined the distribution of neurons and terminals immunoreactive for glutamic acid decarboxylase (GAD) in the thalamus and adjacent structures of the opossum (Didelphis virginiana) and the rabbit and have compared this distribution with the distributions we described previously for the cat and bushbaby (Galago senegalensis). The significance of these experiments depends, first, on the fact that GAD is the synthetic enzyme for GABA, and therefore that GAD immunoreactivity is a marker for GABAergic inhibitory neurons, and second, on previous findings that suggest that GABAergic neurons in the dorsal thalamus are local circuit neurons. In both cat and Galago, GAD-immunoreactive neurons are distributed essentially throughout the entire thalamus. In the opossum, GAD neurons are chiefly confined to the dorsal lateral geniculate nucleus and the lateral extremity of the lateral posterior nucleus. The distribution of GAD neurons in the rabbit is intermediate between that found in the opossum on the one hand and cat and Galago on the other. Like opossum, about 25% of the neurons in the lateral geniculate nucleus of rabbit are GAD immunoreactive. Unlike opossum, however, as many as 18% of the cells in the ventral posterior nucleus of the rabbit are GAD immunoreactive, and scattered cells are also labeled in other thalamic areas, such as the medial geniculate and the lateral group. Aside from the findings in the dorsal thalamus, the chief observation is that GAD-immunoreactive neurons and/or terminals densely fill all principal targets of the optic tract, including the ventral lateral geniculate nucleus; the superficial gray layer of the superior colliculus; the anterior, posterior, and olivary pretectal nuclei; the nucleus of the optic tract; and the medial and lateral terminal nuclei of the accessory optic tract. These results support the idea first put forward by Cajal that local circuit neurons increase in number during the course of the evolution of complex mammalian brains. If we can assume that the conservative opossum retains characteristics reflecting an early stage of mammalian evolution, the results suggest that thalamic local circuit neurons arose first in the visual system and only later in evolution spread throughout the thalamus. PMID:6090511

Penny, G R; Conley, M; Schmechel, D E; Diamond, I T

1984-09-01

308

Pituitary adenylate cyclase activating polypeptide immunoreactivity in the rat spinal cord and medulla: implication of sensory and autonomic functions.  

UK PubMed Central (United Kingdom)

Immunoreactivity to pituitary adenylate cyclase activating polypeptide-38 was detected in numerous nerve fibres in layers I and II of the dorsal horn of the rat and some of these fibres extended into the deeper layers of all segments of the spinal cord. Immunoreactivity was also detected in the lateral funiculus projecting into the intermediolateral cell column of the lower cervical and thoracic segments and in the lateral pathway terminating in the intermediate gray area of the lower lumbar and sacral segments. Neurons in the lateral horn area were not immunoreactive nor were the ventral horn motoneurons. In the medulla, numerous immunoreactive fibres were observed in the spinal trigeminal tract and superficial layers of the caudal spinal trigeminal nucleus but few in the interpolar spinal trigeminal nucleus. A prominent immunoreactive nerve bundle emanated from the caudal spinal trigeminal nucleus and projected into the solitary tract. A dense network of immunoreactive neurons and fibres was present in the nucleus raphe obscurus, lateral reticular nucleus and parvocellular lateral reticular nucleus. Immunoreactive fibres could also be detected in the solitary tract and area postrema. Labelled somata were occasionally noted in various subnuclei of the nucleus of the solitary tract and nucleus raphe pallidus. In addition, a small number of positive neurons were detected in an area between the lateral reticular nucleus and inferior olive and near the ventral surface of the medulla (parapyramidal region). A few weakly-labelled cells were occasionally seen in the dorsal motor nucleus of vagus. A population of neurons in the trigeminal, nodose and dorsal root ganglia from all segments of the spinal cord displayed low to intense immunoreactivity. The presence of immunoreactivity in nodose and dorsal root ganglia, dorsal horn, spinal autonomic nuclei, solitary tract and in certain areas of the medulla suggests that this peptide may participate in a variety of sensory and autonomic functions.

Dun NJ; Miyazaki T; Tang H; Dun EC

1996-08-01

309

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

1985-01-01

310

Identification of immunoreactive proteins of Streptococcus agalactiae isolated from cultured tilapia in China.  

UK PubMed Central (United Kingdom)

Streptococcus agalactiae (Group B streptococcus, GBS) is an important zoonotic pathogen which can cause lethal infections in humans and animals, including aquatic species. Immunoreactive proteins of the S. agalactiae strain, GD201008-001, isolated from cultured tilapia in China, were screened by immunoproteomics using hyperimmune sera, convalescent guinea pig sera and GD201008-001-infected tilapia antisera as primary detection antibodies. A total of 16 different proteins were identified including 13 novel immunoreactive proteins of S. agalactiae. Four proteins, serine-rich repeat glycoprotein 1, branched-chain alpha-keto acid dehydrogenase subunit E2, 5'-nucleotidase family protein and ornithine carbamoyltransferase, were shown to react with the three types of sera and thus were considered to represent novel S. agalactiae vaccine candidate antigens. Our findings represent the basis for vaccine development for piscine S. agalactiae and are necessary for understanding virulence factors and immunogenicity of S. agalactiae with different hosts. This article is protected by copyright. All rights reserved.

Liu G; Zhang W; Lu C

2013-08-01

311

Fos-like immunoreactivity in rat hippocampal neurons following transient global ischemia.  

UK PubMed Central (United Kingdom)

Immediate early genes are expressed following ischemia in many tissues including the brain. Using a chest compression global ischemia model that produced delayed neuronal degeneration in surviving rats, we examined the hippocampal Fos response to ischemia/reperfusion by immunohistochemistry and electron microscopy. Immunostained nuclei were seen in a few CA1 pyramidal cells 1-3 h after reperfusion while the entire dentate granular cell population was immunoreactive. By electron microscopy, subcellular Fos-like immunoreactive sites were found both in the cell nuclei and on segments of endoplasmic reticulum. These findings indicate that transient global ischemia differentially affects the early response genes in neurons of the hippocampal subfields and that such difference may be related to the adult neuroplasticity of the brain.

Tseng MT; Chan SA; Reid KH; Iyer VG

1997-02-01

312

Fos-like immunoreactivity in rat hippocampal neurons following transient global ischemia.  

Science.gov (United States)

Immediate early genes are expressed following ischemia in many tissues including the brain. Using a chest compression global ischemia model that produced delayed neuronal degeneration in surviving rats, we examined the hippocampal Fos response to ischemia/reperfusion by immunohistochemistry and electron microscopy. Immunostained nuclei were seen in a few CA1 pyramidal cells 1-3 h after reperfusion while the entire dentate granular cell population was immunoreactive. By electron microscopy, subcellular Fos-like immunoreactive sites were found both in the cell nuclei and on segments of endoplasmic reticulum. These findings indicate that transient global ischemia differentially affects the early response genes in neurons of the hippocampal subfields and that such difference may be related to the adult neuroplasticity of the brain. PMID:9090643

Tseng, M T; Chan, S A; Reid, K H; Iyer, V G

1997-02-01

313

Involvement of a serotonergic control in the regulation of plasma levels of immunoreactive beta-endorphin.  

UK PubMed Central (United Kingdom)

Beta-endorphin immunoreactivity was measured in plasma and hypothalamus of rats treated with alpha-methyl-p-tyrosine (alpha -MT) (two doses of 200 mg/kg) or p-chloro-phenyl-alanine (PCPA) (two doses of 150 mg/kg). It was also measured in plasma after a single dose (5 mg/kg) of amphetamine or after electrical stimulation of median raphe nucleus (MRN). Plasma levels of immunoreactive beta-endorphin (ir B-E) were significantly decreased by PCPA and were elevated by electrical stimulation of MRN. Alpha -MT was ineffective to modify ir B-E plasma concentration as well as amphetamine. These findings suggest a role for 5-hydroxytryptamine (5-HT) in the regulation of plasma B-E content.

Cancela LM; Fulginiti S; Ramírez OA

1985-01-01

314

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(OH)2) 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.

Greenlee Justin J; Nicholson Eric M; Hamir Amir N; Noyes Gary P; Holtzapple Mark T; Kehrli Marcus E

2008-01-01

315

Two missense mutations causing tyrosinemia type 1 with presence and absence of immunoreactive fumarylacetoacetase.  

UK PubMed Central (United Kingdom)

Hereditary tyrosinemia type 1, due to a deficiency of fumarylacetoacetase (FAH), is characterized by progressive liver damage and renal tubular dysfunction and may occur in an acute or a chronic form. An Ala 134 to Asp (GCT to GAT) transition was found in one Turkish and two Norwegian patients with chronic tyrosinemia. SphI digestion of polymerase chain reaction (PCR) amplified genomic DNA identified the mutation and showed that the patients were heterozygous. All these patients had immunoreactive FAH protein in fibroblasts. Another Norwegian patient with chronic disease, without FAH immunoreactive material in fibroblasts, had a Pro 342 to Leu mutation (CCG to CTG). This mutation was identified by MspI digestion of PCR amplified genomic DNA, and the patient was heterozygous. Northern blotting showed FAH mRNA of normal size and amounts in all patients. Site directed mutagenesis and translation in a rabbit reticulocyte lysate demonstrated that both mutations abolished FAH activity.

Rootwelt H; Chou J; Gahl WA; Berger R; Co?kun T; Brodtkorb E; Kvittingen EA

1994-06-01

316

Mapping of serotonin-immunoreactive neurons of Anastrepha obliqua Macquart larvae  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english Serotonin-immunoreactive neurons were identified in the central nervous system (CNS) of Anastrepha obliqua Macquart, 1835 wandering stage larvae. The PAP immunocytochemical method was applied to the entire CNS (whole mounts). About 90 neurons were visualized in the CNS (20 in the brain and 70 in the ventral ganglion). Both somata and axons were strongly stained. These neurons showed a segmental arrangement and bilateral symmetry. All processes presented a basic projection (more) pattern, in which the major fibres travel contralaterally. Comparison of these neurons with serotonergic neurons described in other insects suggests order-specific traits such as cerebral clusters and presence of only one 5-HT immunoreactive neuron in the 8th abdominal neuromere as well.

Boleli, Isabel Cristina; Paulino-Simőes, Zilá Luz

1999-01-01

317

Tracheal cryopreservation: caspase-3 immunoreactivity in tracheal epithelium and in mixed glands  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english Cryopreservation has an immunomodulating effect on tracheal tissue as a result of class II antigen depletion due to epithelium exfoliation. However, not all epithelium is detached. We evaluated the role of apoptosis in the remaining epithelium of 30 cryopreserved tracheal grafts. Caspase-3 immunoreactivity of tracheal epithelium was studied in canine tracheal segments cryopreserved with F12K medium, with or without subsequent storage in liquid nitrogen at -196°C for 15 d (more) ays. Loss of structural integrity of tracheal mixed glands was observed in all cryopreserved tracheal segments. Caspase-3 immunoreactivity in tracheal mucosa and in mixed glands was significantly decreased, in contrast to the control group and to cryopreserved tracheal segments in which it remained high, due to the effect of storage in liquid nitrogen (P

Sotres-Vega, A.; Baltazares-Lipp, M.; Villalba-Caloca, J.; Gaxiola-Gaxiola, M.O.; Santibańez-Salgado, J.A.; Olmos-Zúńiga, J.R.; Jasso-Victoria, R.

2009-12-01

318

Mapping of serotonin-immunoreactive neurons of Anastrepha obliqua Macquart larvae  

Directory of Open Access Journals (Sweden)

Full Text Available Serotonin-immunoreactive neurons were identified in the central nervous system (CNS) of Anastrepha obliqua Macquart, 1835 wandering stage larvae. The PAP immunocytochemical method was applied to the entire CNS (whole mounts). About 90 neurons were visualized in the CNS (20 in the brain and 70 in the ventral ganglion). Both somata and axons were strongly stained. These neurons showed a segmental arrangement and bilateral symmetry. All processes presented a basic projection pattern, in which the major fibres travel contralaterally. Comparison of these neurons with serotonergic neurons described in other insects suggests order-specific traits such as cerebral clusters and presence of only one 5-HT immunoreactive neuron in the 8th abdominal neuromere as well.

Isabel Cristina Boleli; Zilá Luz Paulino-Simőes

1999-01-01

319

Abnormal Returns and Contrarian Strategies  

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Full Text Available We test the hypothesis that strategies which are long on portfolios of looser stocks and short on portfolios of winner stocks generate abnormal returns in Brazil. This type of evidence for the US stock market was interpreted by The Bondt and Thaler (1985) as reflecting systematic evaluation mistakes caused by investors overreaction to news related to the firm performance. We found evidence of contrarian strategies profitability for horizons from 3 months to 3 years in a sample of stock returns from BOVESPA and SOMA from 1986 to 2000. The strategies are more profitable for shorter horizons. Therefore, there was no trace of the momentum effect found by Jagadeesh and Titman (1993) for the same horizons with US data. There are remaing unexplained positive returns for contrarian strategies after accounting for risk, size, and liquidity. We also found that the strategy profitability is reduced after the Real Plan, which suggests that the Brazilian stock market became more efficient after inflation stabilization.

Marco Bonomo; Ivana Dall'Agnol

2003-01-01

320

Microvascular abnormalities in Rett syndrome.  

UK PubMed Central (United Kingdom)

Rett syndrome (RTT) is a post-natal neurological disorder that represents the second most common cause for mental retardation. The presence of cold hands and feet, and blue, a feature frequently observed in these patients, is one of the non-neurological phenotypes that characterizes RTT, up to now not well explained. We have performed videocapillaroscopy in subjects affected by Rett syndrome. We have observed ramified and bushy capillaries, characteristic features of neoangiogenic capillaries, dilated capillaries and an irregular and chaotic microvascular pattern. To quantify these features and to evaluate the microvascular pattern complexity, we have performed a fractal analysis. Fractal dimension and Lempel-Ziv indexes resulted higher in Rett females than in age-matched healthy females (p < 0.001; p < 0.001). Our findings indicate the presence of previously unrecognized microvascular abnormalities in Rett syndrome.

Bianciardi G; Acampa M; Lamberti I; Sartini S; Servi M; Biagi F; Bocchi V; Hayek J; Pastorelli M

2013-01-01

 
 
 
 
321

Congenital Abnormalities and Multiple Sclerosis  

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Full Text Available Abstract Background There is a strong maternal parent-of-origin effect in determining susceptibility to multiple sclerosis (MS). One hypothesis is that an abnormal intrauterine milieu leading to impaired fetal development could plausibly also result in increased susceptibility to MS. A possible marker for this intrauterine insult is the presence of a non-fatal congenital anomaly. Methods We investigated whether or not congenital anomalies are associated with MS in a population-based cohort. We identified 7063 MS index cases and 2655 spousal controls with congenital anomaly information from the Canadian Collaborative Project on Genetic Susceptibility to MS (CCPGSMS). Results The frequency of congential anomalies were compared between index cases and controls. No significant differences were found. Conclusions Congenital anomalies thus do not appear to be associated with MS. However, we did not have complete data on types and severity of congenital anomalies or on maternal birth history and thus this study should be regarded as preliminary.

Ramagopalan Sreeram V; Guimond Colleen; Criscuoli Maria; Dyment David A; Orton Sarah-Michelle; Yee Irene M; Ebers George C; Sadovnick Dessa

2010-01-01

322

Mammaglobin and S-100 immunoreactivity in salivary gland carcinomas other than mammary analogue secretory carcinoma.  

Science.gov (United States)

Mammary analogue secretory carcinoma (MASC) is a recently described salivary gland tumor that has morphologic features similar to secretory carcinoma of the breast and that also harbors the same ETV6 translocation. Diffuse mammaglobin and S-100 immunoreactivity are used to differentiate MASC from its morphologic mimics, especially acinic cell carcinoma and adenocarcinoma, not otherwise specified. However, the combination of mammaglobin and S-100 immunoreactivity has not been well studied in other types of salivary gland carcinomas that may have focal areas reminiscent of MASC. Here we evaluated mammaglobin and S-100 immunoreactivity in 15 cases each of polymorphous low-grade adenocarcinoma, adenoid cystic carcinoma and mucoepidermoid carcinoma, and also in 2 cases of adenocarcinoma, not otherwise specified, and 1 mucinous adenocarcinoma. Cases with significant co-expression of mammaglobin and S-100 (moderate or strong immunoreactivity in >25% of tumor cells) were further analyzed by fluorescence in situ hybridization using the ETV6 (12p13) break-apart probe. Nine cases (60%) of polymorphous low-grade adenocarcinoma and two (13.3%) of adenoid cystic carcinoma met the criteria for significant co-expression of mammaglobin and S-100. All were negative for the ETV6 translocation by fluorescence in situ hybridization. Although mammaglobin and S-100 positivity was seen in the majority of polymorphous low-grade adenocarcinomas and a minority of adenoid cystic carcinomas, none were positive for the ETV6 translocation characteristic of MASC. This indicates a need for caution in the use of immunohistochemistry for diagnosing MASC, especially in the absence of cytogenetic confirmation. PMID:24029710

Patel, Kalyani R; Solomon, Isaac H; El-Mofty, Samir K; Lewis, James S; Chernock, Rebecca D

2013-09-10

323

Localization of met-enkephalin like immunoreactivity in the glabrous skin of the cat rhinarium.  

UK PubMed Central (United Kingdom)

The presence of met-enkephalin like immunoreactivity (MEL IR) was investigated immunohisto-chemically in the glabrous skin of the cat rhinarium using the peroxidase-antiperoxidase Sternberger's method. Neither sensory corpuscles nor nerve bundles show MEL IR. MEL IR was found in the epidermal Merkel cells, as well as in Langerhans cells and/or melanocytes. In dermal papillae the reaction results positive in a number of cells which could be identified as Schwann or pigmentary cells.

Vega JA; Hernández LC; Del Valle ME; Dubovy P; Bengoechea ME; Pérez-Casas A

1990-01-01

324

Mammaglobin and S-100 immunoreactivity in salivary gland carcinomas other than mammary analogue secretory carcinoma.  

UK PubMed Central (United Kingdom)

Mammary analogue secretory carcinoma (MASC) is a recently described salivary gland tumor that has morphologic features similar to secretory carcinoma of the breast and that also harbors the same ETV6 translocation. Diffuse mammaglobin and S-100 immunoreactivity are used to differentiate MASC from its morphologic mimics, especially acinic cell carcinoma and adenocarcinoma, not otherwise specified. However, the combination of mammaglobin and S-100 immunoreactivity has not been well studied in other types of salivary gland carcinomas that may have focal areas reminiscent of MASC. Here we evaluated mammaglobin and S-100 immunoreactivity in 15 cases each of polymorphous low-grade adenocarcinoma, adenoid cystic carcinoma and mucoepidermoid carcinoma, and also in 2 cases of adenocarcinoma, not otherwise specified, and 1 mucinous adenocarcinoma. Cases with significant co-expression of mammaglobin and S-100 (moderate or strong immunoreactivity in >25% of tumor cells) were further analyzed by fluorescence in situ hybridization using the ETV6 (12p13) break-apart probe. Nine cases (60%) of polymorphous low-grade adenocarcinoma and two (13.3%) of adenoid cystic carcinoma met the criteria for significant co-expression of mammaglobin and S-100. All were negative for the ETV6 translocation by fluorescence in situ hybridization. Although mammaglobin and S-100 positivity was seen in the majority of polymorphous low-grade adenocarcinomas and a minority of adenoid cystic carcinomas, none were positive for the ETV6 translocation characteristic of MASC. This indicates a need for caution in the use of immunohistochemistry for diagnosing MASC, especially in the absence of cytogenetic confirmation.

Patel KR; Solomon IH; El-Mofty SK; Lewis JS Jr; Chernock RD

2013-09-01

325

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

UK PubMed Central (United Kingdom)

Astrocytes express two sodium-coupled transporters, GLAST and GLT-1, which are essential for the maintenance of low extracellular glutamate levels. Here, we performed a comparative analysis of the laminar and subcellular expression profile of GLAST and GLT-1 in the developing postnatal mouse hippocampus using immunohistochemistry and western blotting and employing high-resolution fluorescence microscopy. Astrocytes were identified by co-staining with GFAP or S100ß. In CA1, the density of GFAP-positive cells and GFAP expression rose during the first two weeks after birth, paralleled by a steady increase in GLAST immunoreactivity and protein content. Up-regulation of GLT-1 was completed only at postnatal days (P) P20-25 and thus delayed by about ten days. GLAST staining was highest along 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 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 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. J. Comp. Neurol., 2013. © 2013 Wiley Periodicals, Inc.

Schreiner AE; Durry S; Aida T; Stock MC; Rüther U; Tanaka K; Rose CR; Kafitz KW

2013-08-01

326

Nerve growth factor receptor immunoreactivity in the new world monkey (Cebus apella) and human cerebellum.  

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The present study used the NGFR-5 monoclonal antibody raised against human nerve growth factor receptor (NGFR) to determine the extent of NGFR immunoreactivity within the embryonic and young adult Cebus apella cerebellum as well as the human cerebellum. Immunohistochemically processed tissue revealed NGFR expressing Purkinje cell somata, axons, and dendrites, the latter being observed within the molecular layer of both adult species. Within all regions of the cerebellum we observed both darkly and lightly immunostained Purkinje cells. The proximal axons of these cells, which were visualized for short distances within the granular cell layer, appeared to contain bulbous aggregates of reaction product. In sagittal sections, the full extent of the Purkinje cell dendritic tree was observed in the more lightly stained portions of the cerebellum. In situ hybridization experiments revealed NGFR mRNA within Purkinje cells in a pattern similar to that seen with immunohistochemistry. The distribution of NGFR immunoreactivity within the cerebellum exhibits a general topographic organization with the heaviest and most consistent staining occurring within the archi- and neocerebellum and weaker staining within the paleocerebellum. In fetal Cebus monkey cerebellum obtained at gestational day 50 and 70, NGFR immunoreactivity was observed as a band composed of developing Purkinje cell neurites. These profiles were seen in the paleo- and neocerebellum, but not the archicerebellum. The present investigation is the first demonstration of NGFR immunoreactive profiles in the adult monkey and human cerebellum. These findings suggest that nerve growth factor may influence locomotor and vestibular behaviors that are mediated by cerebellar circuity. The precise mode of action for the NGF/NGFR system within the cerebellum remains to be determined. PMID:1650799

Mufson, E J; Higgins, G A; Kordower, J H

1991-06-22

327

Nerve growth factor receptor immunoreactivity in the new world monkey (Cebus apella) and human cerebellum.  

UK PubMed Central (United Kingdom)

The present study used the NGFR-5 monoclonal antibody raised against human nerve growth factor receptor (NGFR) to determine the extent of NGFR immunoreactivity within the embryonic and young adult Cebus apella cerebellum as well as the human cerebellum. Immunohistochemically processed tissue revealed NGFR expressing Purkinje cell somata, axons, and dendrites, the latter being observed within the molecular layer of both adult species. Within all regions of the cerebellum we observed both darkly and lightly immunostained Purkinje cells. The proximal axons of these cells, which were visualized for short distances within the granular cell layer, appeared to contain bulbous aggregates of reaction product. In sagittal sections, the full extent of the Purkinje cell dendritic tree was observed in the more lightly stained portions of the cerebellum. In situ hybridization experiments revealed NGFR mRNA within Purkinje cells in a pattern similar to that seen with immunohistochemistry. The distribution of NGFR immunoreactivity within the cerebellum exhibits a general topographic organization with the heaviest and most consistent staining occurring within the archi- and neocerebellum and weaker staining within the paleocerebellum. In fetal Cebus monkey cerebellum obtained at gestational day 50 and 70, NGFR immunoreactivity was observed as a band composed of developing Purkinje cell neurites. These profiles were seen in the paleo- and neocerebellum, but not the archicerebellum. The present investigation is the first demonstration of NGFR immunoreactive profiles in the adult monkey and human cerebellum. These findings suggest that nerve growth factor may influence locomotor and vestibular behaviors that are mediated by cerebellar circuity. The precise mode of action for the NGF/NGFR system within the cerebellum remains to be determined.

Mufson EJ; Higgins GA; Kordower JH

1991-06-01

328

Abnormal iron homeostasis and neurodegeneration  

Science.gov (United States)

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 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.

Muhoberac, Barry B.; Vidal, Ruben

2013-01-01

329

Abnormal iron homeostasis and neurodegeneration.  

Science.gov (United States)

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 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. PMID:23908629

Muhoberac, Barry B; Vidal, Ruben

2013-07-30

330

Abnormal iron homeostasis and neurodegeneration.  

UK PubMed Central (United Kingdom)

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 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.

Muhoberac BB; Vidal R

2013-01-01

331

Na,K-ATPase ? Subunit-like Immunoreactive Proteins in Human Body Fluids  

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Full Text Available Na,K-ATPase was recently considered a target for breast cancer treatment since its potent inhibitors ouabain and related digitalis compounds that were reported circulating in body fluids before, were found to possess potent anti-breast cancer activity. Based on this finding, it is hypothesized that Na,K-ATPase derivatives could be circulating in the body and potential candidate biomarkers for early disease detection. Western blotting analysis was employed to investigate the possible presence of Na,K-ATPase-like proteins in cerebrospinal fluid (CSF) and plasma using previously prepared antisera against Na,K-ATPase α�subunit and its formic acid-derived fragments. Results from this analysis show that human Na,K-ATPase α��subunit-like immunoreactive proteins with apparent molecular weights of 97, 68, 66, 57 and 55 kDa in the CSF, as well as 68, 66, 57 and 55 kDa in the plasma have been identified. By combining immunoaffininty binding and electroelution, these immunoreactive proteins were purified from human body fluids. This is the first report of the presence in the body fluids of immunoreactive proteins cross-reacted with anti-human Na,K-ATPase catalytic α subunit antibodies. These results suggest that these proteins are probably Na,K-ATPase derivatives resulting from degradation and released in vivo during cellular metabolism.

F. Peng Jeng-Hsiung

2006-01-01

332

Protein gene product 9.5-immunoreactive nerve fibres and cells in human skin.  

UK PubMed Central (United Kingdom)

Sections of human skin were processed according to the indirect immunofluorescence technique with a rabbit antiserum against human protein gene product 9.5 (PGP 9.5). Immunoreactivity was detected in intraepidermal and dermal nerve fibres and cells. The intraepidermal nerves were varicose or smooth with different diameters, running as single processes or branched, straight or bent, projecting in various directions and terminating in the stratum basale, spinosum or granulosum. The density of the intraepidermal nerves varied between the different skin areas investigated. PGP 9.5-containing axons of the lower dermis were found in large bundles. They separated into smaller axon bundles within the upper dermis, entering this portion of the skin perpendicular to the surface. Then they branched into fibres mainly arranged parallel to the epidermal-dermal junctional zone. However, the fibres en route to the epidermis traversed the upper dermis more or less perpendicularly. Furthermore, immunoreactive dermal nerve fibres were found in the Meissner corpuscles, the arrector pili muscles, hair follicles, around the eccrine and apocrine sweat glands and around certain blood vessels. Such fibres were also observed around most subcutaneous blood vessels, sometimes heavily innervating these structures. Numerous weakly-to-strongly PGP 9.5-immunoreactive cells were found both in the epidermis and in the dermis.

Wang L; Hilliges M; Jernberg T; Wiegleb-Edström D; Johansson O

1990-07-01

333

Immunoreactivity of thymosin beta 4 in human foetal and adult genitourinary tract  

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Full Text Available Thymosins beta 4 (T?4) is a member of the beta-thymosins family, a family of peptides playing essential roles in many cellular functions. Our recent studies suggested T?4 plays a key role in the development of human salivary glands and the gastrointestinal tract. The aim of this study was to analyse the presence of T?4 in the human adult and foetal genitourinary tract. Immunolocalization of T?4 was studied in autoptic samples of kidney, bladder, uterus, ovary, testicle and prostate obtained from four human foetuses and four adults. Presence of the peptide was observed in cells of different origin: in surface epithelium, in gland epithelial cells and in the interstitial cells. T?4 was mainly found in adult and foetal bladder in the transitional epithelial cells; in the adult endometrium, glands and stromal cells were immunoreactive for the peptide; T?4 was mainly localized in the glands of foetal prostate while, in the adults a weak T?4 reactivity was restricted to the stroma. In adult and foetal kidney, T?4 reactivity was restricted to ducts and tubules with completely spared glomeruli; a weak positivity was observed in adult and foetal oocytes; immunoreactivity was mainly localized in the interstitial cells of foetal and adult testis. In this study, we confirm that T?4 could play a relevant role during human development, even in the genitourinary tract, and reveal that immunoreactivity for this peptide may change during postnatal and adult life.

S. Nemolato; T. Cabras; M.U. Fanari; F. Cau; D. Fanni; C. Gerosa; B. Manconi; I. Messana; M. Castagnola; G. Faa

2010-01-01

334

Technetium-99m-labeled monoclonal antibody with preserved immunoreactivity and high in vivo stability  

International Nuclear Information System (INIS)

Recent availability of monoclonal antibodies (MoAb) and their radiolabeling through the use of the bifunctional chelating agents (BCA) have become an alternative procedure for in vivo radioimmunodetection. Using a newly synthesized BCA, a p-carboxyethylphenylglyoxal-di(N-methylthiosemicarbazone) (CE-DTS), the coupling and technetium-99m (/sup 99m/Tc) labeling of monoclonal IgG against hCG were carried out. In the system presented, factors affecting stability and immunoreactivity were examined. Immunoreactivity of the original IgG (56C) was preserved by conjugating one CE-DTS molecule per molecule of IgG (56C) using the phosphorylazide method, however, /sup 99m/Tc labeling pH affected the immunoreactivity and limited the /sup 99m/Tc labeling reaction between pH 4.5 and 6.2. A screening of labeling conditions, such as pH, reaction time, and reducing agent system were then carried out. Technetium-99m-labeled IgG (56C), [/sup 99m/Tc]CE-DTS-IgG (56C), showed good stability upon incubation with mice sera and comparable mice biodistribution to that of indium-111 (111In) DTPA-IgG (56C). Thus, these results indicate the excellent potential of CE-DTS as a BCA for labeling MoAb with /sup 99m/Tc

1987-01-01

335

Technetium-99m-labeled monoclonal antibody with preserved immunoreactivity and high in vivo stability  

Energy Technology Data Exchange (ETDEWEB)

Recent availability of monoclonal antibodies (MoAb) and their radiolabeling through the use of the bifunctional chelating agents (BCA) have become an alternative procedure for in vivo radioimmunodetection. Using a newly synthesized BCA, a p-carboxyethylphenylglyoxal-di(N-methylthiosemicarbazone) (CE-DTS), the coupling and technetium-99m (/sup 99m/Tc) labeling of monoclonal IgG against hCG were carried out. In the system presented, factors affecting stability and immunoreactivity were examined. Immunoreactivity of the original IgG (56C) was preserved by conjugating one CE-DTS molecule per molecule of IgG (56C) using the phosphorylazide method, however, /sup 99m/Tc labeling pH affected the immunoreactivity and limited the /sup 99m/Tc labeling reaction between pH 4.5 and 6.2. A screening of labeling conditions, such as pH, reaction time, and reducing agent system were then carried out. Technetium-99m-labeled IgG (56C), (/sup 99m/Tc)CE-DTS-IgG (56C), showed good stability upon incubation with mice sera and comparable mice biodistribution to that of indium-111 (/sup 111/In) DTPA-IgG (56C). Thus, these results indicate the excellent potential of CE-DTS as a BCA for labeling MoAb with /sup 99m/Tc.

Arano, Y.; Yokoyama, A.; Furukawa, T.; Horiuchi, K.; Yahata, T.; Saji, H.; Sakahara, H.; Nakashima, T.; Koizumi, M.; Endo, K.

1987-06-01

336

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

Science.gov (United States)

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

337

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

UK PubMed Central (United Kingdom)

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.

Dubbeldam JL; den Boer-Visser AM

2000-04-01

338

Usefulness of cytokeratin immunoreactivity pattern for distinction of Barrett's esophagus from intestinal metaplasia of the stomach.  

UK PubMed Central (United Kingdom)

The histological distinction between Barrett's esophagus involving the distal esophagus and intestinal metaplasia of the stomach has important clinical implications and can be difficult even with the use of histochemical stains. Cytokeratin (CK) 7 and 20 are cytoplastic structural proteins that show restricted expression in normal and malignant epithelia of the gastrointestinal tract. CK7 and 20 immunostaining were performed on a 67-year-old male with cardiac cancer with reflux esophagitis due to sliding hernia. The CK7/20 immunoreactivity pattern of cancer and reflux esophagitis in this case showed superficial CK20 staining and strong CK7 staining of both superficial and deep glands. In intestinal metaplasia of the stomach, strong CK20 immunostaining in superficial and deep glands and absent CK7 immunoreactivity were noted. Neither CK7 nor CK20 immunoreactivity was noted in squamous cell epithelium. Therefore, we concluded that in this patient intestinal metaplasia of the esophagus was BE. The CK7/20 reactivity pattern is useful for identifying the intestinal metaplasia of the esophagus from the stomach using histological materials from biopsy and surgically resected specimens.

Iwata T; Kurita N; Nishioka M; Hidenori M; Wakatsuki S; Sano T; Shimada M

2007-09-01

339

Usefulness of cytokeratin immunoreactivity pattern for distinction of Barrett's esophagus from intestinal metaplasia of the stomach.  

Science.gov (United States)

The histological distinction between Barrett's esophagus involving the distal esophagus and intestinal metaplasia of the stomach has important clinical implications and can be difficult even with the use of histochemical stains. Cytokeratin (CK) 7 and 20 are cytoplastic structural proteins that show restricted expression in normal and malignant epithelia of the gastrointestinal tract. CK7 and 20 immunostaining were performed on a 67-year-old male with cardiac cancer with reflux esophagitis due to sliding hernia. The CK7/20 immunoreactivity pattern of cancer and reflux esophagitis in this case showed superficial CK20 staining and strong CK7 staining of both superficial and deep glands. In intestinal metaplasia of the stomach, strong CK20 immunostaining in superficial and deep glands and absent CK7 immunoreactivity were noted. Neither CK7 nor CK20 immunoreactivity was noted in squamous cell epithelium. Therefore, we concluded that in this patient intestinal metaplasia of the esophagus was BE. The CK7/20 reactivity pattern is useful for identifying the intestinal metaplasia of the esophagus from the stomach using histological materials from biopsy and surgically resected specimens. PMID:18019700

Iwata, Takashi; Kurita, Nobuhiro; Nishioka, Masanori; Hidenori, Miyamoto; Wakatsuki, Shingo; Sano, Toshiaki; Shimada, Mitsuo

2007-09-01

340

Two peptidergic drugs increase the synaptophysin immunoreactivity in brains of 6-week-old rats.  

Science.gov (United States)

An increase of synaptic density has been found in the hippocampus, the dendate gyrus and in the entorhinal cortex of 6-week-old rats after 7 days of treatment with the peptidergic drug Cerebrolysin, its peptide preparation E021 and the diluted peptide preparation E021dil. Rats received drugs on postnatal days 1-7 (2.5 ml/kg, each day). Controls received saline. The animals were sacrificed on days 42-48 of their life, after they had undergone behavioural testing in a Morris water maze. Slices of brain were stained immunohistochemically with anti-synaptophysin, a specific marker of presynaptic terminals. The synaptophysin-immunoreactivity of presynaptic terminals was quantified using light microscopy and a computerised image analysis system. Our results showed that rats benefit from the treatment with both drugs. A significant increase in the number of synaptophysin-immunoreactive presynaptic terminals was found in the entorhinal cortex and the hippocampal subfields CA1, CA2, CA3 stratum radiatum and CA3 stratum lucidum. The increased immunoreactive presynaptic terminals found in the present study are in accordance with the positive effects of the drugs on spatial learning and memory in young rats (Gschanes & Windisch 1999). PMID:10816071

Windholz, E; Gschanes, A; Windisch, M; Fachbach, G

2000-02-01

 
 
 
 
341

Two peptidergic drugs increase the synaptophysin immunoreactivity in brains of 6-week-old rats.  

UK PubMed Central (United Kingdom)

An increase of synaptic density has been found in the hippocampus, the dendate gyrus and in the entorhinal cortex of 6-week-old rats after 7 days of treatment with the peptidergic drug Cerebrolysin, its peptide preparation E021 and the diluted peptide preparation E021dil. Rats received drugs on postnatal days 1-7 (2.5 ml/kg, each day). Controls received saline. The animals were sacrificed on days 42-48 of their life, after they had undergone behavioural testing in a Morris water maze. Slices of brain were stained immunohistochemically with anti-synaptophysin, a specific marker of presynaptic terminals. The synaptophysin-immunoreactivity of presynaptic terminals was quantified using light microscopy and a computerised image analysis system. Our results showed that rats benefit from the treatment with both drugs. A significant increase in the number of synaptophysin-immunoreactive presynaptic terminals was found in the entorhinal cortex and the hippocampal subfields CA1, CA2, CA3 stratum radiatum and CA3 stratum lucidum. The increased immunoreactive presynaptic terminals found in the present study are in accordance with the positive effects of the drugs on spatial learning and memory in young rats (Gschanes & Windisch 1999).

Windholz E; Gschanes A; Windisch M; Fachbach G

2000-02-01

342

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

International Nuclear Information System (INIS)

[en] 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

343

Alteration in NCX-3 immunoreactivity within the gerbil hippocampus following spontaneous seizures.  

UK PubMed Central (United Kingdom)

Although NCX-3 is highly expressed in the brain, the distribution of NCX-3 in the epileptic hippocampus is still controversial. Therefore, to assess the distribution and pattern of NCX-3 expression in epileptic hippocampus, we performed a comparative analysis of NCX-3 immunoreactivities in the hippocampus of seizure-resistant (SR) and seizure-sensitive (SS) gerbils. In SR gerbils, NCX-3 immunoreactivity was higher than pre-seizure SS gerbils, particularly in the pavalbumin (PV)-positive interneurons. Three h post-ictal, NCX-3 immunoreactivity in the SS gerbil hippocampus was markedly elevated to the level of SR gerbils. Six h post-ictal, the expression of NCX-3 was reduced to the level of pre-seizure SS gerbils. Therefore, the results of the present study suggest that down-regulation of NCX-3 expression in the SS gerbil hippocampus may be involved in the hyperexcitability of SS gerbils due to an imbalance of intracellular Na(+)/Ca(2+) homeostasis and Ca(2+) concentration.

Park DK; Park KH; Ko JS; Kim DS

2011-05-01

344

FBXO7 immunoreactivity in ?-synuclein-containing inclusions in Parkinson disease and multiple system atrophy.  

Science.gov (United States)

Mutations in the gene encoding the F-box only protein 7 (FBXO7) cause PARK15, an autosomal recessive form of juvenile parkinsonism. Although the brain pathology in PARK15 patients remains unexplored, in vivo imaging displays severe loss of nigrostriatal dopaminergic terminals. Understanding the pathogenesis of PARK15 might therefore illuminate the mechanisms of the selective dopaminergic neuronal degeneration, which could also be important for understanding idiopathic Parkinson disease (PD). The expression of FBXO7 in the human brain remains poorly characterized, and its expression in idiopathic PD and different neurodegenerative diseases has not been investigated. Here, we studied FBXO7 protein expression in brain samples of normal controls (n = 9) and from patients with PD (n = 13), multiple system atrophy (MSA) (n = 5), Alzheimer disease (AD) (n = 5), and progressive supranuclear palsy (PSP) (n = 5) using immunohistochemistry with 2 anti-FBXO7 antibodies. We detected widespread brain FBXO7 immunoreactivity, with the highest levels in neurons of the cerebral cortex, putamen, and cerebellum. There were no major differences between normal and PD brains overall, but FBXO7 immunoreactivity was detected in large proportions of ?-synuclein-positive inclusions (Lewy bodies, Lewy neurites, glial cytoplasmic inclusions), where it colocalized with ?-synuclein in PD and MSA cases. By contrast, weak FBXO7 immunoreactivity was occasionally detected in tau-positive inclusions in AD and PSP. These findings suggest a role for FBXO7 in the pathogenesis of the synucleinopathies. PMID:23656991

Zhao, Tianna; Severijnen, Lies-Anne; van der Weiden, Marcel; Zheng, Ping Pin; Oostra, Ben A; Hukema, Renate K; Willemsen, Rob; Kros, Johan M; Bonifati, Vincenzo

2013-06-01

345

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

2012-11-19

346

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

UK PubMed Central (United Kingdom)

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.

Zhang F; Li J; Zou M; Chen Y; Wang Y; Qi X

2013-04-01

347

Tracheal cryopreservation: caspase-3 immunoreactivity in tracheal epithelium and in mixed glands  

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Full Text Available Cryopreservation has an immunomodulating effect on tracheal tissue as a result of class II antigen depletion due to epithelium exfoliation. However, not all epithelium is detached. We evaluated the role of apoptosis in the remaining epithelium of 30 cryopreserved tracheal grafts. Caspase-3 immunoreactivity of tracheal epithelium was studied in canine tracheal segments cryopreserved with F12K medium, with or without subsequent storage in liquid nitrogen at -196°C for 15 days. Loss of structural integrity of tracheal mixed glands was observed in all cryopreserved tracheal segments. Caspase-3 immunoreactivity in tracheal mucosa and in mixed glands was significantly decreased, in contrast to the control group and to cryopreserved tracheal segments in which it remained high, due to the effect of storage in liquid nitrogen (P < 0.05, ANOVA and Tukey test). We conclude that apoptosis can be triggered in epithelial cells during tracheal graft harvesting even prior to cryopreservation, and although the epithelial caspase-3 immunoreactivity is reduced in tracheal cryopreservation, this could be explained by increased cell death. Apoptosis cannot be stopped during tracheal cryopreservation.

A. Sotres-Vega; M. Baltazares-Lipp; J. Villalba-Caloca; M.O. Gaxiola-Gaxiola; J.A. Santibańez-Salgado; J.R. Olmos-Zúńiga; R. Jasso-Victoria

2009-01-01

348

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; Loviselli Andrea

2006-01-01

349

Lysosomal hydrolases of different classes are abnormally distributed in brains of patients with Alzheimer disease.  

UK PubMed Central (United Kingdom)

beta-Amyloid formation requires multiple abnormal proteolytic cleavages of amyloid precursor protein (APP), including one within its intramembrane domain. Lysosomes, which contain a wide variety of proteases (cathepsins) and other acid hydrolases, are major sites for the turnover of membrane proteins and other cell constituents. Using immunocytochemistry, immunoelectron microscopy, and enzyme histochemistry, we studied the expression and cellular distributions of 10 lysosomal hydrolases, including 4 cathepsins, in neocortex from patients with Alzheimer disease and control (non-Alzheimer-disease) individuals. In control brains, acid hydrolases were localized exclusively to intracellular lysosome-related compartments, and 8 of the 10 enzymes predominated in neurons. In Alzheimer disease brains, strongly immunoreactive lysosomes and lipofuscin granules accumulated markedly in the perikarya and proximal dendrites of many cortical neurons, some of which were undergoing degeneration. More strikingly, these same hydrolases were present in equally high or higher levels in senile plaques in Alzheimer disease, but they were not found extracellularly in control brains, including those from Parkinson or Huntington disease patients. At the ultrastructural level, hydrolase immunoreactivity in senile plaques was localized to extracellular lipofuscin granules similar in morphology to those within degenerating neurons. Two cathepsins that were undetectable in neurons were absent from senile plaques. These results show that lysosome function is altered in cortical neurons in Alzheimer disease. The presence of a broad spectrum of acid hydrolases in senile plaques indicates that lysosomes and their contents may be liberated from cells, principally neurons and their processes, as they degenerate. Because cathepsins can cleave polypeptide sites on APP relevant for beta-amyloid formation, their abnormal extracellular localization and dysregulation in Alzheimer disease can account for the multiple hydrolytic events in beta-amyloid formation. The actions of membrane-degrading acid hydrolases could also explain how the intramembrane portion of APP containing the C terminus of beta-amyloid becomes accessible to proteases.

Cataldo AM; Paskevich PA; Kominami E; Nixon RA

1991-12-01

350

Lysosomal hydrolases of different classes are abnormally distributed in brains of patients with Alzheimer disease.  

Science.gov (United States)

beta-Amyloid formation requires multiple abnormal proteolytic cleavages of amyloid precursor protein (APP), including one within its intramembrane domain. Lysosomes, which contain a wide variety of proteases (cathepsins) and other acid hydrolases, are major sites for the turnover of membrane proteins and other cell constituents. Using immunocytochemistry, immunoelectron microscopy, and enzyme histochemistry, we studied the expression and cellular distributions of 10 lysosomal hydrolases, including 4 cathepsins, in neocortex from patients with Alzheimer disease and control (non-Alzheimer-disease) individuals. In control brains, acid hydrolases were localized exclusively to intracellular lysosome-related compartments, and 8 of the 10 enzymes predominated in neurons. In Alzheimer disease brains, strongly immunoreactive lysosomes and lipofuscin granules accumulated markedly in the perikarya and proximal dendrites of many cortical neurons, some of which were undergoing degeneration. More strikingly, these same hydrolases were present in equally high or higher levels in senile plaques in Alzheimer disease, but they were not found extracellularly in control brains, including those from Parkinson or Huntington disease patients. At the ultrastructural level, hydrolase immunoreactivity in senile plaques was localized to extracellular lipofuscin granules similar in morphology to those within degenerating neurons. Two cathepsins that were undetectable in neurons were absent from senile plaques. These results show that lysosome function is altered in cortical neurons in Alzheimer disease. The presence of a broad spectrum of acid hydrolases in senile plaques indicates that lysosomes and their contents may be liberated from cells, principally neurons and their processes, as they degenerate. Because cathepsins can cleave polypeptide sites on APP relevant for beta-amyloid formation, their abnormal extracellular localization and dysregulation in Alzheimer disease can account for the multiple hydrolytic events in beta-amyloid formation. The actions of membrane-degrading acid hydrolases could also explain how the intramembrane portion of APP containing the C terminus of beta-amyloid becomes accessible to proteases. PMID:1837142

Cataldo, A M; Paskevich, P A; Kominami, E; Nixon, R A

1991-12-15

351

Clinicopathologic features of autosomal recessive amyotrophic lateral sclerosis associated with optineurin mutation.  

UK PubMed Central (United Kingdom)

We performed clinicopathological analyses of two amyotrophic lateral sclerosis (ALS) patients with homozygous Q398X?optineurin (OPTN) mutation. Clinically, both patients presented signs of upper and lower motor neuron degeneration, but only Patient 1 showed gradual frontal dysfunction and extrapyramidal signs, and temporal lobe and motor cortex atrophy. Neuropathological examination of Patient 1 revealed extensive cortical and spinal motor neuron degeneration and widespread degeneration of the basal ganglia. Bilateral corticospinal tracts exhibited degeneration. Loss of spinal anterior horn cells (AHCs) and gliosis were observed, whereas posterior columns, Clarke's columns, intermediate lateral columns, and the Onuf's nucleus were spared. In the brainstem, moderate neuronal loss and gliosis were noted in the hypoglossal and facial motor nuclei. No Bunina bodies were found in the surviving spinal and brainstem motor neurons. Transactivation response (TAR) DNA-binding protein 43 (TDP-43)-positive neuronal and glial cytoplasmic inclusions were observed throughout the central nervous system. The Golgi apparatus in motor neurons of the brainstem and spinal cord was often fragmented. Immunoreactivity for OPTN was not observed in the brain and spinal cord, consistent with nonsense-mediated mRNA decay of OPTN. The TDP-43 pathology of Q398X was similar to that of an autosomal dominant E478G mutation. This result suggests that the loss-of-function, but not the proteinopathy itself, of OPTN results in TDP-43 deposits in neuronal and glial cytoplasm and Golgi apparatus fragmentation, leading to multisystem neurodegeneration.

Kamada M; Izumi Y; Ayaki T; Nakamura M; Kagawa S; Kudo E; Sako W; Maruyama H; Nishida Y; Kawakami H; Ito H; Kaji R

2013-07-01

352

Neuropeptide Y-like immunoreactivity in rat cranial parasympathetic neurons: coexistence with vasoactive intestinal peptide and choline acetyltransferase  

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Neuropeptide Y (NPY) is widely distributed in the sympathetic nervous system, where it is colocalized with norepinephrine. The authors report here that NPY-immunoreactive neurons are also abundant in three cranial parasympathetic ganglia, the otic, sphenopalatine, and ciliary, in the rat measured by radioimmunoassay. High-performance liquid chromatographic analysis of the immunoreactive material present in the otic ganglion indicates that this material is very similar to porcine NPY and indistinguishable from the NPY-like immunoreactivity present in rat sympathetic neurons. These findings raise the possibility that NPY acts as a neuromodulator in the parasympathetic as well as the sympathetic nervous system. In contrast to what had been observed for sympathetic neurons, NPY-immunoreactive neurons in cranial parasympathetic ganglia do not contain detectable catecholamines or tyrosine hydroxylase immunoreactivity, and many do contain immunoreactivity for vasoactive intestinal peptide and/or choline acetyltransferase. These findings suggest that there is no simple rule governing coexpression of NPY with norepinephrine, acetylcholine, or vasoactive intestinal peptide in autonomic neurons. Further, while functional studies have indicated that NPY exerts actions on the peripheral vasculature which are antagonistic to those of acetylcholine and vasoactive intestinal peptide, the present results raise the possibility that these three substances may have complementary effects on other target tissues.

Leblanc, G.C.; Trimmer, B.A.; Landis, S.C.

1987-05-01

353

Apoptosis, cell proliferation and serotonin immunoreactivity in gut of Liza aurata from natural heavy metal polluted environments: preliminary observations  

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Full Text Available In the present paper, the effect of natural environment nonlethal heavy metal concentration on cell renewal of Liza aurata intestinal epithelium, was studied by the TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling) method and anti-PCNA (proliferating cell nuclear antigen) immunohistochemistry, in order to detect, respectively, apoptosis and cell proliferation. In addition, the presence and distribution of the cell renewal regulator, serotonin, was immunohistochemically investigated. In order to reduce variability, only immature specimens were considered. The results indicated that in the control specimens from non-polluted areas, the PCNA immunoreactive nuclei of the proximal intestinal epithelium were only located at the bottom of the intestinal folds, together with a few TUNEL-positive nuclei, and goblet mucous differentiated cells. In the specimens from polluted areas, the number of PCNA immunoreactive cells was greatly enhanced, and they extended along the mid portion of the intestinal folds; the number of TUNEL-positive nuclei was enhanced as well, but they were almost exclusively detected in the third apical portion of the intestinal folds. Serotonin immunoreactive nerve elements were more frequently detected in the intestinal wall of L. aurata specimens from polluted areas, and besides that, some serotonin immunoreactive endocrine cells were also present. Variations in distribution and frequency of TUNEL-positive nuclei, PCNA immunoreactive nuclei, and serotonin immunoreactivity put in evidence an alteration of cell renewal with an enhancement of cell proliferation, probably leading to morphological intestinal fold changes.

S Ferrando; T Ferrando; L Girosi; A Mauceri; S Fasulo; G Tagliafierro

2005-01-01

354

Odontodysplasia, gingival manifestations, and accompanying abnormalities.  

UK PubMed Central (United Kingdom)

Regional odontodysplasia is an uncommon developmental dental disorder that may occasionally be accompanied by other abnormalities. A case is described in which the chief report was of a gingival enlargement arising in a female patient who also had dolichocephaly, thin calvarium, clinodactyly and transverse grooving of her fingernails, and a history of abnormal hair. Previously suggested etiologic factors and cases reported in association with other abnormalities are reviewed.

Fanibunda KB; Soames JV

1996-01-01

355

Odontodysplasia, gingival manifestations, and accompanying abnormalities.  

Science.gov (United States)

Regional odontodysplasia is an uncommon developmental dental disorder that may occasionally be accompanied by other abnormalities. A case is described in which the chief report was of a gingival enlargement arising in a female patient who also had dolichocephaly, thin calvarium, clinodactyly and transverse grooving of her fingernails, and a history of abnormal hair. Previously suggested etiologic factors and cases reported in association with other abnormalities are reviewed. PMID:8850490

Fanibunda, K B; Soames, J V

1996-01-01

356

Abnormal fat distribution in PMM2-CDG.  

UK PubMed Central (United Kingdom)

We hypothesize that abnormal fat distribution, a common feature of PMM2-CDG, is associated with abnormal perinatal hormone regulation. We assessed 32 cases with PMM2-CDG, for the comorbidity of hypoglycemia/hyperinsulinism and fat pads. Ninety percent of patients with hypoketotic hypoglycemia and/or hyperinsulinism had abnormal fat distribution, while normoglycemic patients showed this feature in 50% of the cases. This statistically significant difference suggests an etiological role of the insulin receptor in developing abnormal fat distribution in PMM2-CDG.

Wolthuis DF; van Asbeck EV; Kozicz T; Morava E

2013-11-01

357

The XXXXY sex chromosome abnormality.  

UK PubMed Central (United Kingdom)

The most common sex chromosome complex in sex chromatin-positive males with Klinefelter's syndrome is XXY. When the complex is XXYY or XXXY, the clinical findings do not seem to differ materially from those seen in XXY subjects, although more patients with these intersexual chromosome complements need to be studied to establish possible phenotypical expressions of the chromosomal variants.Two male children with an XXXXY sex chromosome abnormality are described. The data obtained from the study of these cases and five others described in the literature suggest that the XXXXY patient is likely to have congenital defects not usually seen in the common form of the Klinefelter syndrome. These include a triad of (1) skeletal anomalies (including radioulnar synostosis), (2) hypogenitalism (hypoplasia of penis and scrotum, incomplete descent of testes and defective prepubertal development of seminiferous tubules), and (3) greater risk of severe mental deficiency.That the conclusions are based on data from a small number of patients is emphasized, together with the need for a cytogenetic survey of a large control or unselected population.

BARR ML; CARR DH; POZSONYI J; WILSON RA; DUNN HG; JACOBSON TS; MILLER JR; LEWIS M; CHOWN B

1962-10-01

358

[Renal abnormalities in ankylosing spondylitis].  

UK PubMed Central (United Kingdom)

We will study the epidemiologic, clinical, biological, therapeutic, prognostic characteristics and predictive factors of development of nephropathy in ankylosing spondylitis patients. We retrospectively reviewed the medical record of 32 cases with renal involvement among 212 cases of ankylosing spondylitis followed in our service during the period spread out between 1978 and 2006. The renal involvement occurred in all patients a mean of 12 years after the clinical onset of the rheumatic disease. Thirty-two patients presented one or more signs of renal involvement: microscopic hematuria in 22 patients, proteinuria in 23 patients, nephrotic syndrome in 11 patients and decreased renal function in 24 patients (75%). Secondary renal amyloidosis (13 patients), which corresponds to a prevalence of 6,1% and tubulointerstitial nephropathy (7 patients) were the most common cause of renal involvement in ankylosing spondylitis followed by IgA nephropathy (4 patients). Seventeen patients evolved to the end stage renal disease after an average time of 29.8 ± 46 months. The average follow-up of the patients was 4,4 years. By comparing the 32 patients presenting a SPA and renal disease to 88 with SPA and without nephropathy, we detected the predictive factors of occurred of nephropathy: tobacco, intense inflammatory syndrome, sacroileite stage 3 or 4 and presence of column bamboo. The finding of 75% of the patients presented a renal failure at the time of the diagnosis of renal involvement suggests that evidence of renal abnormality involvement should be actively sought in this disease.

Samia B; Hazgui F; Abdelghani KB; Hamida FB; Goucha R; Hedri H; Taarit CB; Maiz HB; Kheder A

2012-07-01

359

[Renal abnormalities in ankylosing spondylitis].  

Science.gov (United States)

We will study the epidemiologic, clinical, biological, therapeutic, prognostic characteristics and predictive factors of development of nephropathy in ankylosing spondylitis patients. We retrospectively reviewed the medical record of 32 cases with renal involvement among 212 cases of ankylosing spondylitis followed in our service during the period spread out between 1978 and 2006. The renal involvement occurred in all patients a mean of 12 years after the clinical onset of the rheumatic disease. Thirty-two patients presented one or more signs of renal involvement: microscopic hematuria in 22 patients, proteinuria in 23 patients, nephrotic syndrome in 11 patients and decreased renal function in 24 patients (75%). Secondary renal amyloidosis (13 patients), which corresponds to a prevalence of 6,1% and tubulointerstitial nephropathy (7 patients) were the most common cause of renal involvement in ankylosing spondylitis followed by IgA nephropathy (4 patients). Seventeen patients evolved to the end stage renal disease after an average time of 29.8 ± 46 months. The average follow-up of the patients was 4,4 years. By comparing the 32 patients presenting a SPA and renal disease to 88 with SPA and without nephropathy, we detected the predictive factors of occurred of nephropathy: tobacco, intense inflammatory syndrome, sacroileite stage 3 or 4 and presence of column bamboo. The finding of 75% of the patients presented a renal failure at the time of the diagnosis of renal involvement suggests that evidence of renal abnormality involvement should be actively sought in this disease. PMID:22520483

Samia, Barbouch; Hazgui, Faiçal; Abdelghani, Khaoula Ben; Hamida, Fethi Ben; Goucha, Rym; Hedri, Hafedh; Taarit, Chokri Ben; Maiz, Hedi Ben; Kheder, Adel

2012-04-18

360

Hemostatic abnormalities in liver cirrhosis  

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Full Text Available In this study, 44 patients with liver cirrhosis were investigated for hemostatic parameters. Patients with spontaneous bacterial peritonitis, hepatocellular carcinoma, hepatorenal syndrome and cholestatic liver diseases were excluded. Patients were classified by Child-Pugh criterion and according to this 4 patients were in Class A, 20 in Class B and 20 in C. Regarding to these results, it was aimed to investigate the haematological disturbances in liver cirrhotic patients.In the result there was a correlation between activated partial thromboplastin time, serum iron, ferritin, transferrin, haptoglobin and Child-Pugh classification. Besides there was no correlation between prothrombin time, factor 8 and 9, protein C and S, anti-thrombin 3, fibrinogen, fibrin degradation products, serum iron binding capacity, hemoglobin, leukocyte, mean corpuscular volume and Child-Pugh classification.There were significant difference, in terms of AST, ferritin, haptoglobulin, sex and presence of ascites between groups (p0.05). In the summary, we have found correlation between hemostatic abnormalities and disease activity and clinical prognosis in patients with liver cirrhosis which is important in the management of these patients. This is also important for identification of liver transplant candidiates earlier.

Kendal YALÇIN; Orhan AYYILDIZ

2009-01-01