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

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

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

Our most significant finding, however, is that a 43-kDa species of TDP-43 is present in FTLD-associated TDP-43 aggregates. The aggregated 43-kDa species migrates in SDS-PAGE identically to the physiologically predominant TDP-43 species, and lacks obvious covalent modifications. Like the physiologically predom- inant species, it is not phosphorylated at the 409/410 sites. Known sites of TDP-43 phosphorylation in FTLD include serines at codons 379, 403, 404, 409 and 410 [18]. Our studies do not conclusively exclude phosphorylation at sites other than 409 and 410. However, we note that a phosphorylated 43-kDa species was not identified in the sarcosyl-insoluble fraction from FTLD or ALS tissue using antibodies against TDP-43 phosphorylation sites at S409, S410, S409/410, S403/404, or S379 [18]. Many types of covalent modifications of protein are known to occur, and some might not alter the migration of the protein in SDS-PAGE [29]. Without large amounts of purified protein it is not possible to exclude most such modifications. Nevertheless, our data suggest that the covalent structure of the 43-kDa species in FTLD- associated aggregates is identical to that of the normally predominant physiological form of brain TDP-43. The mass-yield of this species in our preparations is small, so it has not been possible to determine its structure by mass spectrometry. Figure 3. The method for analyzing components of aggregates isolated on SDD-AGE gels. (A) An unstained large format SDD-AGE gel after a typical preparative electrophoretic run. The dashed boxes depict the regions that were excised for concentration and analysis of proteins. The upper box labeled ‘‘A’’ circumscribes the ‘‘aggregate region’’ in which the major portion of aggregated TDP-43 associated with FTLD-TDP is found. The lower box labeled M contains the presumably monomeric TDP-43 forms found in both controls and FTLD cases (‘‘monomer region’’). The wide dark band at the bottom of the gel is bromphenol blue
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Dement. neuropsychol.  vol.11 número3

Dement. neuropsychol. vol.11 número3

ABSTRACT. Introduction: TDP-43 is an intranuclear protein involved in many cellular processes. When altered, it shows a change in pattern of distribution, as well as in functioning, throughout the Central Nervous System structures. Frontotemporal Lobar Degeneration (FTLD) and Amyotrophic Lateral Sclerosis (ALS) are examples of TDP-43 proteinopathy. These disorders form a clinical spectrum, with some patients having a pure cognitive disorder while others also exhibit motor features. Methods: We studied two donated brains from patients with a diagnosis of Frontotemporal Dementia (FTD), one of which was associated with ALS (ALS-FTD). After fixation and macroscopic examinations, sample analyses were performed. Specific regions were chosen for the application of immunohistochemistry (IHC) with anti-Aβ, AT8, anti-α-synuclein and anti-phospho-TDP-43. Results: Both brains presented anti-phospho-TDP-43 positivity, but this was not equally distributed throughout the encephalic zones. In the FTD case, the studied brain presented phosphorylated TDP-43- in the frontal cortex, hippocampus, entorhinal cortex and mesencephalon; in the ALS-FTD case, the abnormal protein was also seen in the pons and medulla oblongata. The brain in the ALS-FTD case presented Aβ and AT8 positivity in the hippocampus and entorhinal cortex (Braak I and II). Discussion: The hypothesis supported by scientific literature that these neurodegenerative diseases can have the same etiology with distinct encephalic region involvement is corroborated by the present study.
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Transcriptomic Changes Due to Cytoplasmic TDP-43 Expression Reveal Dysregulation of Histone Transcripts and Nuclear Chromatin.

Transcriptomic Changes Due to Cytoplasmic TDP-43 Expression Reveal Dysregulation of Histone Transcripts and Nuclear Chromatin.

TAR DNA-binding protein 43 (TDP-43) is normally a nuclear RNA-binding protein that exhibits a range of functions including regulation of alternative splicing, RNA trafficking, and RNA stability. However, in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with TDP-43 inclusions (FTLD-TDP), TDP-43 is abnormally phosphorylated, ubiquitinated, and cleaved, and is mislocalized to the cytoplasm where it forms distinctive aggregates. We previously developed a mouse model expressing human TDP-43 with a mutation in its nuclear localization signal (ΔNLS-hTDP-43) so that the protein preferentially localizes to the cytoplasm. These mice did not exhibit a significant number of cytoplasmic aggregates, but did display dramatic changes in gene expression as measured by microar- ray, suggesting that cytoplasmic TDP-43 may be associated with a toxic gain-of-function. Here, we analyze new RNA-sequencing data from the ΔNLS-hTDP-43 mouse model, together with published RNA-sequencing data obtained previously from TDP-43 antisense oligonucleotide (ASO) knockdown mice to investigate further the dysregulation of gene expression in the ΔNLS model. This analysis reveals that the transcriptomic effects of the overexpression of the ΔNLS-hTDP-43 transgene are likely due to a gain of cytoplasmic function. Moreover, cytoplasmic TDP-43 expression alters transcripts that regulate chroma- tin assembly, the nucleolus, lysosomal function, and histone 3’ untranslated region (UTR) processing. These transcriptomic alterations correlate with observed histologic abnormali- ties in heterochromatin structure and nuclear size in transgenic mouse and human brains.
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Inhibition of TDP-43 aggregation by nucleic acid binding.

Inhibition of TDP-43 aggregation by nucleic acid binding.

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 (TDP 208–414 ). Consistently, these two mutants form aggregates, in
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Análise prognóstica da imunoexpressão de osteopontina no microambiente dos carcinomas mamários esporádicos de cadelas

Análise prognóstica da imunoexpressão de osteopontina no microambiente dos carcinomas mamários esporádicos de cadelas

Several proteins have been identified to be differentially expressed in large groups of benign and malignant tumors, and immunohistochemical analysis of biomarkers has a key role in this evaluation. Previous studies on the supplementation of the histopathologic prognosis of the clinical outcome of canine mammary tumors showed the need for the establishment of immunohistochemical markers capable of providing useful data (Klopfleisch et al., 2010). In humans, osteopontin (OPN) – a secreted glycophosphoprotein that has been implicated in a number of different normal physiologic and pathologic processes (Tuck et al., 2007) such as increase in metastatic potential (Kadar et al, 2002) – was demonstrated to be expressed in a variety of tumors (Brown et al. 1994; Coppola et al., 2004). The mechanism of OPN overexpression and its role in human breast cancer is not clear but it was established that OPN interacts with various cell surface receptors such as EGF receptor (Yarden and Sliwkowski, 2001). EGF receptor subfamily genes encode 4 structurally related tyrosine kinase receptors (TKR): epidermal growth factor receptor (ERBB1 gene, EGFR/HER1 protein); ERBB2 gene, erbB2/Neu/HER2 protein; ERBB3 gene, HER3 protein; and ERBB4 gene, HER4 protein, all of them belonging to the type I TKR class. They play an important role in mediating growth factor signaling (Yarden and Sliwkowski, 2001).
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Dysplastic nevus associated with seborrheic keratosis

Dysplastic nevus associated with seborrheic keratosis

present criteria determining it as melanocytic, the remaining lesion characteristics, such as bluish veil and granulation justified excisional biopsy. In the his- tological study, comedo-like opening corresponded to invagination of the epidermis filled by keratin and milia-like cysts to small intra-epidermal cyst filled with keratin. 4 The nests of nevus cells did not present

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Estratégias de aprendizagem em língua estrangeira: investigando a prática da sala de aula

Estratégias de aprendizagem em língua estrangeira: investigando a prática da sala de aula

Nestes termos, de que as estratégias facilitam, aceleram, motivam e direcionam o aprendizado/aquisição de uma nova língua, e ainda, paralelamente, auxiliam na construç[r]

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A PROTO-HISTÓRIA DOS DIREITOS HUMANOS FUNDAMENTAIS

A PROTO-HISTÓRIA DOS DIREITOS HUMANOS FUNDAMENTAIS

Em sentido parecido é a lição de Klaus Stern citado por Sarlet, que sintetiza o devir histórico dos direitos humanos fundamentais até o seu reconhecimento nas primeiras constituições escritas, da seguinte forma: “a) uma pré-história, que se estende até o século XVI; b) uma fase intermediária, aqui corresponde ao período de elaboração da doutrina jusnaturalista e da afirmação dos direitos naturais do homem; c) a fase da constitucionalização, iniciada em 1776, com as sucessivas declarações de direitos dos novos Estados americanos.” (STERN, Klaus. Das staatsrecht der bundes republik deustchland. vol. III/1. Munchen: C. H. Beck, 1984, p. 56 apud SARLET, Ingo Wolfgang. Ob. cit., 2008, p. 43).
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Arq. NeuroPsiquiatr.  vol.59 número1

Arq. NeuroPsiquiatr. vol.59 número1

Neuropsycological examination was limited by poor cooperation. However, the positive findings showed a behaviour that was negativist and irritable, impulsive and inappropiate, with an exaggerated and melodramatic emotional display. His executive-functions such as planning, sequencing, and judgement were compromised. The patient did not initiate conversation, and his responses were brief and often elicited only by continual encouragement. He was fully disoriented regarding both time and space. The clinical findings suggested that he did not have true amnesia, but there was relative preservation of recognition memory compared to free recall. This refleted a general retrieval deficit with difficulty retrieving established memories wi- thout a temporal gradient, that is, there was equal impairment across all life periods. He had retention of concrete material in the face of short-term memory deficits. Lexical retrieval was also profoundly impaired. Language showed spontaneous speech with stereotyped phrases. He did not show very formal difficulty in comprehension, al- though his interpretation of the Token Test indicated severe aphasia of the global type. There was many word finding difficulty and severe anomia. He could not read or write. He had great difficulty carrying out mental calculations. Per- ceptual and spatial skills were not intact, as measured by his poor ability to identify successive groups of objects and block constructions.
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Paradigmenwechsel bei der Alzheimer-Krankheit: Intrazelluläre Aggregation von Amyloid-beta verursacht den Zellverlust unabhängig von extrazellulären Plaques

Paradigmenwechsel bei der Alzheimer-Krankheit: Intrazelluläre Aggregation von Amyloid-beta verursacht den Zellverlust unabhängig von extrazellulären Plaques

Wir konnten zeigen, dass APP/PS1KI-Mäuse einen schwer- wiegenden neurologischen Phänotyp aufweisen, der unter an- derem durch Gewichtsverlust, verzögertes Wachstum, Gang- störungen und die Entwicklung einer Kyphose, die eine spezi- elle Wirbelsäulendeformation darstellt, gekennzeichnet ist [27]. Dieser Phänotyp resultiert vermutlich aus einer progres- siven motorischen Dysfunktion, die auf eine altersabhängige axonale Degeneration in Hirn und Rückenmark zurückzufüh- ren ist. Darüber hinaus beginnt bei 6 Monate alten APP/ PS1KI-Mäusen eine altersabhängige Abnahme des Arbeits- gedächtnisses, was mit einer Reihe unterschiedlicher Verhal- tenstests gezeigt werden konnte. Diese Tests waren so ge- wählt, dass beginnende motorische Defizite keinen Einfluss darstellten [27]. Beeinträchtigungen des Arbeitsgedächtnisses konnten zuvor auch in anderen transgenen Alzheimer-Maus- modellen wie z. B. TgCRND8-Mäusen [40] oder den kürzlich publizierten 5xFAD-Mäusen gezeigt werden, die bereits im Alter von 4–5 Monaten eine deutliche Beeinträchtigung im sogenannten „Y-Maze“, einem Y-förmigen Labyrinth, zeigen. Dies ist von besonderem Interesse, da diese Mäuse ebenfalls hohe Spiegel intrazellulär akkumulierter Aβ-Peptide aufwei- sen [15]. Die Abnahme der Arbeitsgedächtnisleistung im APP/PS1KI-Modell zeigt eine deutliche Koinzidenz mit dem Beginn des Nervenzellverlustes in der CA1-Region des Hippokampus im Alter von 6 Monaten, die sich zu einem 50%-igen Zellverlust in dieser Region im Alter von 10 Mona- ten ausweitet [22].
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Retinoic acid protects from experimental cerebral infarction by upregulating GAP-43 expression

Retinoic acid protects from experimental cerebral infarction by upregulating GAP-43 expression

respectively. The primers were designed and synthesized by Takara biotechnology Co., Ltd. (China). The results were normalized by the ratio of the target’s relative concentration to that of GAPDH in the same sample. Real-time PCR cycles were conducted for the amplification of GAP-43 and GAPDH cDNA using a thermal cycler (MJ Research Com- pany, DFC-3200, USA). Real-time PCR was conducted using 2 mL of cDNA in a final reaction volume of 20 mL according to the protocol of the Real Master Mix kit (Tiangen Biotech Co., Ltd., China). The PCR cycling program was set for 1 cycle of pre-denaturation at 95°C for 90 s, then 39 cycles at 95°C for 10 s, 60°C for 30 s, and an 80°C plate reading for 3 s; melting curve data were acquired at every 0.2°C from 55°C to 95°C, and there was a 1-s hold between reads. Because the target and refer- ence (glyceraldehyde 3-phosphate dehydrogenase, GAPDH) gene have different sequences and amplicon lengths, it is probable that they show different PCR efficiencies. Results are reported as the target/reference ratio.
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The recombinant protein rSP03B is a valid antigen for screening dog exposure to Phlebotomus perniciosus across foci of canine leishmaniasis

The recombinant protein rSP03B is a valid antigen for screening dog exposure to Phlebotomus perniciosus across foci of canine leishmaniasis

This study follows the canine longitudinal study conducted in southern Italy by Kostalova et al. (2015), which described the dynamics and diagnostic potential of antibodies recogniz- ing Phlebotomus perniciosus (Larroussius subgenus) salivary recombinant proteins in dogs following natural exposure to sandflies over 2 years. Factors such as salivary antigens, age and expected sandfly dynamics were considered as variables and were therefore carefully evaluated. The most reactive and reproducible antigen was found to be the 43-kDa yellow-related recombinant protein (rSP03B) from P. perniciosus saliva. In view of these promising results, the rSP03B antigen was tested in canine sera samples collected cross-sectionally in canine leish- maniasis (CanL) endemic settings in Italy and Portugal. The study evaluated levels of individual canine antigenic responses to P. perniciosus rSP03B compared with P. perniciosus SGH, and the degree of similarity in these antigenic associations, across endemic canine populations in Portuguese and Italian foci in order to assess the universal use of rSP03B as a marker of natural sandfly exposure. Previous research had confirmed the presence of two native yellow-related proteins in P. perniciosus salivary gland transcriptome and proteome (Anderson et al., 2006). Therefore, the antigenic similarity of rSP03B to its native form was studied and the specificity of the anti-rSP03B immunoglobulin G (IgG) antibody response was confirmed by the inclusion of 42-kDa yellow-related recombinant protein (rSP03).
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Análise da presença de mutação no gene TARDBP em pacientes com degeneração lobar...

Análise da presença de mutação no gene TARDBP em pacientes com degeneração lobar...

Brazil is one of the developing countries that are undergoing a process of demographic transition in which the elderly represents a significant proportion of the total population. Neurodegenerative illnesses most commonly appear at such ages. Frontotemporal lobar degeneration (FTLD) and Alzheimer ’s disease (AD) are the most frequent causes for dementia. The investigation of risk factors for degenerative dementia is a relevant subject of neurosciences and the evaluation of the nature of genetic risk factors has produced the most important contributions. Mutations in TARDBP gene, the encoder of the TDP-43 nuclear protein, appear as the most frequent genetic occurrences for FTLD, whereas, in DA, the ε4 allele of the apolipoprotein E (APOE) is the major genetic risk factor. Patients with clinical diagnosis of FTLD types of families and sporadic (n=47) and probable AD (n=30) from the ambulatory of Cognitive Neurology Group and Behavior (CNGB) of Neurological Clinic of HC-FMUSP were invited to participate in this study. Blood samples were collected for lymphocytic DNA extraction. The APOE gene polymorphisms are being determined through the real time PCR technique. The 1-6 exons of TARDBP gene were amplified by PCR and their products were sequenced in automated sequencer. The TDP-43 gene analysis in patients with FTLD showed the presence of one mutation in the region of exon 6 TARDBP gene in a male patient of 54 years old, with diagnoses of semantic dementia. Regarding DA patients genotyping, the real time methodology has been shown as an efficient approach to detect APOE polymorphisms, presenting data similar to those observed in other Brazilian studies.
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Arnopheles darlingi is a mosquito species closely associated with man in the

Arnopheles darlingi is a mosquito species closely associated with man in the

Seis isoenzimas de esterase foram estudadas durante o desenvolvimento de Anopheles darlingi usando eletroforese em gel de poliacrilamida e dois substratos diferentes,.. a-naftilacetato e[r]

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Experimental host range of Citrus leprosis virus C (CiLV-C)

Experimental host range of Citrus leprosis virus C (CiLV-C)

relationship is of the circulative type (Kitajima & Alberti, 2010b). The entire CiLV-C genome has been sequenced (Locali-Fabris et al., 2006; Pascon et al., 2006), and is distinct from those of other known viruses. Thus it was placed in the new genus Cilevirus (Locali-Fabris et al., 2012). Symptoms of infection are localized lesions on leaves, fruits and stems (Rodrigues et al., 2003; Bastianel et al., 2010). For a long time, CiLV-C was considered to be restricted to Citrus spp., with sweet oranges considered highly susceptible, mandarins (C. reticulata Blanco) and grapefruits (C. paradisi Macfad.) moderately susceptible, and lemons [C. limon (L.) Osbeck] practically immune (Bastianel et al., 2010). However, mechanical transmission assays have demonstrated that some herbaceous hosts, such as Chenopodium quinoa Willd., C. amaranticolor H.J. Coste & A. Reyn. and Gomphrena globosa L. are susceptible to CiLV-C, responding to exposure with localized lesions (Colariccio et al., 1995). The first case of natural infection of a non-Citrus plant by CiLV-C was found in Swinglea glutinosa (Blanco) Merr. (Rutaceae), used in hedgerow around citrus orchards in Villavicenzo, Colombia (León et al., 2008). More recently, Commelina benghalensis L. (Commelinaceae) plants growing spontaneously in an organic sweet orange orchard in Borborema, SP, Brazil, were found to be naturally infested with B. phoenicis and infected by CiLV-C (Nunes et al., 2012a). Experimental
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當今美國圖書館學校之解析 An Analysis of American Library Association Accredited Degree Programs

當今美國圖書館學校之解析 An Analysis of American Library Association Accredited Degree Programs

University of Washington 的 Information School 提供學位較多樣化 ,計有下列五 種:Bachelors of Science in Information;Master of Library and Information Science; Master of Library and Information Science—Law;Master of Science in Information Management;Ph.D in Information Science。其成立的宗旨在培養資訊專業人才 ,包 括圖書館學與資訊科學 ,課程強調人與知識連結的重要性以及資訊的取得是基本人 權。課程主要分成三大領域: ㈠資訊與生活品質;㈡人們的資訊能量;㈢以人為中 心的資訊系統與服務的設計。資訊 、人與技術是三大領域之共同核心元素 (註 4) 。
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Concepções e repercuções da avaliação da aprendizagem no ensino jurídico: uma análise a partir da visão dos atores envolvidos no processo

Concepções e repercuções da avaliação da aprendizagem no ensino jurídico: uma análise a partir da visão dos atores envolvidos no processo

and students view of the UFOPA law school (remaining Class UFPA / Campus of Santarém). Conceptualized to learning assessment analysis in legal education and applied a questionnaire to investigated, in order to collect detailed data on the questioning under study. We opted for the literature review, which supported and guided the analysis of the study, aiming at a systematic interpretation. It was observed that the peculiarities of the traditional teaching are perpetuated and that the manifestations of changes were solely curricular nature. Graphics print the opinion of students and teachers about the experienced teaching-learning process, importance of evaluation, to know the assessment of changes or adjustments to traditional methods, techniques and teaching resources, clarification of the objectives of the tests applied, and returning the evaluation result. We conclude that, although the views of teachers and students are conflicting [in part] to overcome the compartmentalized and fragmented knowledge is as it requires face to interdisciplinarity as a tool in the construction of a new pedagogical approach to legal education.
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O olho da barbárie, Marildo Menegat

O olho da barbárie, Marildo Menegat

No artigo Civilização em excesso será usada exatamente essa expressão, retirada do Manifesto Comunista, para demonstrar como a dinâmica avassaladora de desenvolvimento das [r]

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Metadata與OAI-PMH在新聞數位典藏之整合應用 Integrated Application of Metadata and OAI-PMH to Digital News Archive

Metadata與OAI-PMH在新聞數位典藏之整合應用 Integrated Application of Metadata and OAI-PMH to Digital News Archive

This paper studies the metadata integration problem between the Taiwan Baseball News Database and the Union Catalog of National Digital Archives Program. First, we design a metadata transformation system which can transform the news articles to four popular metadata formats: DC, NITF, RSS and DAC. The system not only supports metadata transformation but also RSS subscription and batch exportation for Union Catalog. Secondly, based on the OAI-PMH architecture, the digital news archives plays a role as Repository, and then we extend the system to be a Data Pro- vider. It can be queried by any Service Provider, parse the Verbs, and then respond the Record Sets to the Service Provider with certain metadata format. As a result, the digital news can be delivered more eficient.
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江南製造局翻譯館翻譯西書方法考 Research on the Method of Western Science Translation in the Translation Department of the Jiangnan Arsenal

江南製造局翻譯館翻譯西書方法考 Research on the Method of Western Science Translation in the Translation Department of the Jiangnan Arsenal

為了避免譯名前後不一的現象 ,江南製造局翻譯館先後編寫出幾種名目表。其 中重要的有「化學材料中西名目表」(英文書名 Vocabulary of Names of Substance of Occurring in Various Words on Chemistry: Chie fly in Bloxam’s Chemistry),「西藥大成 藥品中西名目表」(英文書名 Vocabulary of Names of Material Medica Occurring in the Translation of Royle’s Manual of Material Medica and Therapeutics with Lists of Name and Places Occurring in the Same Work, and in Various Treatise on Chemistry and Allied Subjects) ,「金石中西名目表」(英文書名 Vocabulary of Mineralogical Terms Occurring in the Manual by J.D.Dana A.M.)和「汽機中西名目表」 (英文書名為 Vocabulary of Terms Relating to the Steam Engine) 。這些譯名對照表 ,雖未在全國範圍內遵行 ,卻 成為規範中西文翻譯的工具 ,對于後來譯名的統一有著很大影響。這也是江南製造 局翻譯館對後世的貢獻之一。
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