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1.
Phytopathology ; 110(1): 80-84, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31631804

RESUMO

The pepper cultivar Yellow Lantern, one of the spiciest pepper varieties, is a local germplasm of Capsicum chinense, cultivated exclusively on Hainan Island, China. However, this variety is susceptible to viral diseases that severely affect its production. In this study, we report that pepper veinal mottle virus (PVMV) is associated with foliar chlorosis and rugosity symptoms in Yellow Lantern. To verify this correlation, we constructed a full-length cDNA clone of a PVMV isolate named HNu. The virus progeny derived from the cDNA clone replicated and moved systemically in the pepper, inducing the same symptoms as those induced by PVMV-HNu in Yellow Lantern peppers in the field. The results support that PVMV-HNu is the causal agent of foliar chlorosis and rugosity disease in Yellow Lantern. This knowledge will help in the diagnosis and prevention of disease caused by PVMV. Furthermore, the cDNA clone serves as a reverse genetic tool to study the molecular pathogenesis of PVMV.


Assuntos
Capsicum , Doenças das Plantas , Potyvirus/genética , Capsicum/virologia , China , Clonagem de Organismos , DNA Complementar/genética , Doenças das Plantas/virologia , Potyvirus/fisiologia
2.
J Virol Methods ; : 113795, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31809783

RESUMO

Two reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed for the detection of areca palm necrotic ringspot virus (ANRSV) and areca palm necrotic spindle-spot virus (ANSSV), respectively. These two emerging viruses both induce necrotic symptoms in areca palms. The coat protein (CP) gene of ANRSV and the 9 K gene of ANSSV were used to design the respective RT-LAMP primers for the assays. Each set of four primers designed for each of these viruses was found to be highly specific in the detection of the respective targeted virus. The optimal incubation conditions for the RT-LAMP assays were 63 °C for 40 min for ANRSV and at 61 °C for 40 min for ANSSV. The sensitivity of the RT-LAMP method for each of these viruses was 10-fold greater than that of the corresponding conventional reverse-transcription polymerase chain reaction (RT-PCR). The RT-LAMP assays may be useful for the rapid early detection of ANSSV and ANRSV in commercial areca palm production.

3.
Annu Rev Virol ; 6(1): 255-274, 2019 09 29.
Artigo em Inglês | MEDLINE | ID: mdl-31299166

RESUMO

Potyviridae is the largest family of plant-infecting RNA viruses, encompassing over 30% of known plant viruses. The family is closely related to animal picornaviruses such as enteroviruses and belongs to the picorna-like supergroup. Like all other picorna-like viruses, potyvirids employ polyprotein processing as a gene expression strategy and have single-stranded, positive-sense RNA genomes, most of which are monopartite with a long open reading frame. The potyvirid polyproteins are highly conserved in the central and carboxy-terminal regions. In contrast, the N-terminal region is hypervariable and contains position-specific mutations resulting from transcriptional slippage during viral replication, leading to translational frameshift to produce additional viral proteins essential for viral infection. Some potyvirids even lack one of the N-terminal proteins P1 or helper component-protease and have a genus-specific or species-specific protein instead. This review summarizes current knowledge about the conserved and divergent features of potyvirid genomes and biological relevance and discusses future research directions.

4.
Arch Virol ; 164(8): 2209-2213, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31161389

RESUMO

The complete genome of a double-stranded RNA (dsRNA) mycovirus, Phoma matteuccicola partitivirus 1 (PmPV1) was sequenced. It consists of two dsRNA segments, 1664 bp (dsRNA-1) and 1383 bp (dsRNA-2) in length, each containing a single open reading frame (ORF) potentially encoding a 46.78-kDa protein and a 40.92-kDa protein, respectively. dsRNA-1 encodes a putative polypeptide with a conserved RNA-dependent RNA polymerase (RdRp) domain that shows sequence similarity to the corresponding proteins of partitiviruses. The protein encoded by dsRNA-2 has no significant similarity to the typical coat proteins (CPs) of partitiviruses, but structure analysis nevertheless suggested that it might function as a coat protein. Purified viral particles of PmPV1 were isometric and approximately 29 nm in diameter. Phylogenetic analysis showed that PmPV1 is closely related to members of the genus Gammapartitivirus within the family Partitiviridae but forms a separate branch with Colletotrichum acutatum RNA virus 1 and Ustilaginoidea virens partitivirus 2. This is the first report of the full-length nucleotide sequence of a novel virus of the genus Gammapartitivirus infecting P. matteuccicola strain LG915, the causal agent of leaf blight of Curcuma wenyujin.


Assuntos
Ascomicetos/virologia , Micovírus/genética , Genoma Viral/genética , Sequência de Aminoácidos , Sequência de Bases , Proteínas do Capsídeo/genética , Curcuma/virologia , Genômica/métodos , Fases de Leitura Aberta/genética , Filogenia , Doenças das Plantas/virologia , RNA Replicase/genética , Vírus de RNA/genética , RNA de Cadeia Dupla/genética , RNA Viral/genética , Análise de Sequência de DNA/métodos
5.
Phytopathology ; 109(5): 887-894, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30133353

RESUMO

Areca palm (Areca catechu), one of the two most important commercial crops in Hainan, China, has been severely damaged by a variety of pathogens and insects. Here, we report a new disease, tentatively referred to as areca palm necrotic ringspot disease (ANRSD), which is highly epidemic in the main growing regions in Hainan. Transmission electron microscopy observation and small RNA deep sequencing revealed the existence of a viral agent of the family Potyviridae in a diseased areca palm plant (XC1). The virus was tentatively named areca palm necrotic ringspot virus (ANRSV). Subsequently, the positive-sense single-stranded genome of ANRSV isolate XC1 was completely determined. The genome annotation revealed the existence of two cysteine proteinases in tandem (HC-Pro1 and HC-Pro2) in the genomic 5' terminus of ANRSV. Sequence comparison and phylogenetic analysis suggested the taxonomic classification of ANRSV into the recently proposed genus Arepavirus in the family Potyviridae. Given the close relationship of ANRSV with another newly reported arepavirus (areca palm necrotic spindle-spot virus), the exact taxonomic status of ANRSV needs to be further investigated. In this study, a reverse transcription polymerase chain reaction assay for ANRSV-specific detection was developed and a close association between ANRSV and ANRSD was found.


Assuntos
Areca/virologia , Filogenia , Doenças das Plantas/virologia , Potyviridae/patogenicidade , China , Genoma Viral , Potyviridae/classificação , RNA Viral
7.
Arch Virol ; 163(12): 3471-3475, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30136252

RESUMO

A novel virus, tentatively named "areca palm necrotic spindle-spot virus" (ANSSV), was identified in Areca catechu L. in Hainan, China, and its complete genomic sequence was determined. Its positive-sense single-stranded RNA genome is comprised of 9,437 nucleotides (nt), excluding the poly (A) tail, and contains one large open reading frame encoding a polyprotein of 3,019 amino acids (aa). A Blastp search showed that the polyprotein of ANSSV shared a maximum of 31%-32% aa sequence identity (with 86%-95% coverage) with all seven known macluraviruses. Nucleotide sequence comparison of the ORF of ANSSV to those of macluraviruses revealed identities ranging from 41.0% to 44.6%, which is less than the inter-genus identity values for the family Potyviridae. Phylogenetic analysis based on either the aa or nt sequence of the polyprotein did not cluster ANSSV into any established or unassigned genus of the family Potyviridae. Therefore, we suggest that ANSSV is the first member of a previously unrecognized genus of the family Potyviridae.


Assuntos
Areca/virologia , Genoma Viral , Doenças das Plantas/virologia , Potyviridae/genética , Potyviridae/isolamento & purificação , Sequência de Bases , China , Dados de Sequência Molecular , Fases de Leitura Aberta , Filogenia , Potyviridae/classificação , Análise de Sequência de DNA
8.
Arch Virol ; 163(9): 2579-2582, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29786122

RESUMO

A macluravirus, tentatively named alpinia oxyphylla mosaic virus (AloMV), was identified in Alpinia oxyphylla, and its complete genomic sequence determined. The positively single-stranded RNA genome is comprised of 8213 nucleotides excluding the poly (A) tail, and contains one large open reading frame encoding a polyprotein of 2,626 amino acids. Blastp search showed that the polyprotein of AloMV shared 48%~68% aa sequence identities with other reported macluraviruses. Phylogenetic analysis based on the nucleotide sequence of the polyprotein showed that AloMV, together with all other macluraviruses, clustered into the same group most closely related to cardamom mosaic virus, sharing 66.3% nt and 68% aa sequence identities, respectively. These data above suggest that AloMV represents an isolate of a putative new member within the genus Macluravirus.


Assuntos
Alpinia/virologia , Genoma Viral , Doenças das Plantas/virologia , Potyviridae/genética , RNA Viral/genética , Proteínas Virais/genética , Sequência de Bases , Mapeamento Cromossômico , Tamanho do Genoma , Fases de Leitura Aberta , Filogenia , Potyviridae/classificação , Potyviridae/isolamento & purificação , Homologia de Sequência do Ácido Nucleico , Sequenciamento Completo do Genoma
9.
Arch Virol ; 163(9): 2583-2586, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29789942

RESUMO

The complete genomic sequence of a telosma mosaic virus (TeMV) isolate (named PasFru), identified in passion fruit in China, was determined. The entire RNA genome of PasFru comprises 10,049 nucleotides (nt) excluding the poly(A) tail and encodes a polyprotein of 3,173 amino acids (aa), flanked by 5' and 3' untranslated regions (UTR) of 276 and 251 nt, respectively. Compared with the previous TeMV isolate Hanoi from Telosma cordata, the only documented isolate with the entire genome sequence annotated, PasFru had an extra 87 nt and 89 aa residues at the 3'-end of 5'UTR and the N-terminus of the P1 protein, respectively, which contributed to the genome size difference between PasFru and Hanoi (10,049 nt versus 9,689 nt). Pairwise sequence comparisons showed that PasFru shares 73.6% nt and 80.9% aa sequence identity with the Hanoi isolate at the whole-genome and polyprotein level, respectively, and these values are below the corresponding threshold values for species demarcation in the family Potyviridae. These data suggest that TeMV-PasFru should be classified as a new member of the genus Potyvirus.


Assuntos
Genoma Viral , Passiflora/virologia , Doenças das Plantas/virologia , Potyviridae/genética , RNA Viral/genética , Proteínas Virais/genética , Sequência de Aminoácidos , Sequência de Bases , Mapeamento Cromossômico , Tamanho do Genoma , Fases de Leitura Aberta , Filogenia , Potyviridae/classificação , Potyviridae/isolamento & purificação , Homologia de Sequência do Ácido Nucleico , Sequenciamento Completo do Genoma
10.
Cancer Manag Res ; 10: 1069-1078, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29780261

RESUMO

Purpose: To report the clinical features, imaging manifestations, histopathology, and immunohistochemical features of several cases of orbital solitary fibrous tumors (SFTs) in a Chinese tertiary hospital, and to undertake a literature review of this rare disease. Methods: A non-comparative retrospective review of clinical presentations, imaging manifestations, histopathology, and immunohistochemical features as well as the management and disease outcomes of patients with orbital SFT was conducted along with a review of orbital SFT cases in the literature. Results: This study includes two male and two female patients, with an average age of 53 years. Common presentations among these patients included a palpable subcutaneous mass, swelling of the eyelid, proptosis, diplopia, and vision disturbance. Three patients (cases 2-4) underwent imaging scans. All patients had complete surgical excisions and the tissue was subjected to pathological analysis. One patient (Case 4) experienced a recurrence with malignant transformation and received a re-excision surgery and postoperative radiotherapy. All patients remain alive and well after a minimum follow-up of 12 months (range 12-34 months). Conclusion: Despite its rare occurrence, we suggest that the possibility of orbital SFTs needs to be considered when a painless, slowly growing orbital mass is identified. Typical characteristic magnetic resonance imaging features of orbital SFTs are iso- or hypointense signals on T1 and T2-weighted images, with marked enhancement. A positive cluster of differentiation 34 (CD34) staining is an important diagnostic clue favoring SFT. Some orbital SFTs are infiltrating, aggressive, or recur with malignant transformation. Therefore, regular long-term follow-up after complete excision is mandatory.

11.
Plant J ; 92(5): 846-861, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28941316

RESUMO

Plasmodesmata (PD), unique to the plant kingdom, are structurally complex microchannels that cross the cell wall to establish symplastic communication between neighbouring cells. Viral intercellular movement occurs through PD. To better understand the involvement of PD in viral infection, we conducted a quantitative proteomic study on the PD-enriched fraction from Nicotiana benthamiana leaves in response to infection by Turnip mosaic virus (TuMV). We report the identification of a total of 1070 PD protein candidates, of which 100 (≥2-fold increase) and 48 (≥2-fold reduction) are significantly differentially accumulated in the PD-enriched fraction, when compared with protein levels in the corresponding healthy control. Among the differentially accumulated PD protein candidates, we show that an α-expansin designated NbEXPA1, a cell wall loosening protein, is PD-specific. TuMV infection downregulates NbEXPA1 mRNA expression and protein accumulation. We further demonstrate that NbEXPA1 is recruited to the viral replication complex via the interaction with NIb, the only RNA-dependent RNA polymerase of TuMV. Silencing of NbEXPA1 inhibits plant growth and TuMV infection, whereas overexpression of NbEXPA1 promotes viral replication and intercellular movement. These data suggest that NbEXPA1 is a host factor for potyviral infection. This study not only generates a PD-proteome dataset that is useful in future studies to expound PD biology and PD-mediated virus-host interactions but also characterizes NbEXPA1 as the first PD-specific cell wall loosening protein and its essential role in potyviral infection.


Assuntos
Doenças das Plantas/virologia , Proteínas de Plantas/metabolismo , Plasmodesmos/metabolismo , Potyvirus/metabolismo , Tabaco/microbiologia , Potyvirus/fisiologia , Proteômica , Tabaco/metabolismo , Replicação Viral
12.
Plant Biotechnol J ; 15(3): 344-356, 2017 03.
Artigo em Inglês | MEDLINE | ID: mdl-27565765

RESUMO

RNA silencing is a powerful technology for molecular characterization of gene functions in plants. A commonly used approach to the induction of RNA silencing is through genetic transformation. A potent alternative is to use a modified viral vector for virus-induced gene silencing (VIGS) to degrade RNA molecules sharing similar nucleotide sequence. Unfortunately, genomic studies in many allogamous woody perennials such as peach are severely hindered because they have a long juvenile period and are recalcitrant to genetic transformation. Here, we report the development of a viral vector derived from Prunus necrotic ringspot virus (PNRSV), a widespread fruit tree virus that is endemic in all Prunus fruit production countries and regions in the world. We show that the modified PNRSV vector, harbouring the sense-orientated target gene sequence of 100-200 bp in length in genomic RNA3, could efficiently trigger the silencing of a transgene or an endogenous gene in the model plant Nicotiana benthamiana. We further demonstrate that the PNRSV-based vector could be manipulated to silence endogenous genes in peach such as eukaryotic translation initiation factor 4E isoform (eIF(iso)4E), a host factor of many potyviruses including Plum pox virus (PPV). Moreover, the eIF(iso)4E-knocked down peach plants were resistant to PPV. This work opens a potential avenue for the control of virus diseases in perennial trees via viral vector-mediated silencing of host factors, and the PNRSV vector may serve as a powerful molecular tool for functional genomic studies of Prunus fruit trees.


Assuntos
Genoma de Planta/genética , Doenças das Plantas/genética , Doenças das Plantas/virologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/virologia , Vírus Eruptivo da Ameixa/patogenicidade , Prunus/genética , Prunus/virologia , Resistência à Doença/genética , Proteínas de Plantas/genética , Interferência de RNA
13.
Onco Targets Ther ; 9: 5281-6, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27601919

RESUMO

PURPOSE: Olfactory neuroblastoma (ONB) is a rare neuroectodermal malignant neoplasm that originates from the olfactory sensory epithelium. In reported cases involving the orbit, the primary neoplasms could invade the orbits through bone destruction or cranio-orbital communication anatomical structure. The present study reports a rare case of orbital metastatic tumor arising from an intracranial ONB in a Chinese patient, which is the first case demonstrating an uncommon route of ONB involving the orbit. PATIENT AND METHODS: A 62-year-old man with a right orbital tumor was diagnosed as having a secondary tumor arising from intracranial ONB. Clinical examination, including ocular imaging examination, and histopathological and immunohistochemical studies were performed. RESULTS: In the ocular examination, a focal, firm, nontender swelling with ill-defined margins in the right orbit was found on palpation. Proptosis, limited eyeball movement, and visual loss were also observed. The unenhanced computed tomography scan showed a homogeneous, isolated, ill-defined soft tissue neoplasm in the right orbit. The histopathological study revealed irregularly shaped nests of tumor cells surrounded by avascularized fibrous stroma. The monomorphic malignant tumor cells are small round blue cells that are slightly larger than mature lymphocytes, with a very high nuclear to cytoplasmic ratio. The immunohistochemical staining for tumor cells was positive for synaptophysin, chromogranin A, cluster of differentiation 56 (CD56), cytokeratin, vimentin, S-100 protein, and negative for epithelial membrane antigen, and cluster of differentiation (CD99). CONCLUSION: This is so far the first Chinese case of orbital metastasis from intracranial ONB that has ever been reported. Metastasis might be another route of ONB invading the orbit besides direct invasion.

14.
J Virol ; 90(10): 5119-5131, 2016 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-26962227

RESUMO

UNLABELLED: The potyviral RNA genome encodes two polyproteins that are proteolytically processed by three viral protease domains into 11 mature proteins. Extensive molecular studies have identified functions for the majority of the viral proteins. For example, 6K2, one of the two smallest potyviral proteins, is an integral membrane protein and induces the endoplasmic reticulum (ER)-originated replication vesicles that target the chloroplast for robust viral replication. However, the functional role of 6K1, the other smallest protein, remains uncharacterized. In this study, we developed a series of recombinant full-length viral cDNA clones derived from a Canadian Plum pox virus (PPV) isolate. We found that deletion of any of the short motifs of 6K1 (each of which ranged from 5 to 13 amino acids), most of the 6K1 sequence (but with the conserved sequence of the cleavage sites being retained), or all of the 6K1 sequence in the PPV infectious clone abolished viral replication. The trans expression of 6K1 or the cis expression of a dislocated 6K1 failed to rescue the loss-of-replication phenotype, suggesting the temporal and spatial requirement of 6K1 for viral replication. Disruption of the N- or C-terminal cleavage site of 6K1, which prevented the release of 6K1 from the polyprotein, either partially or completely inhibited viral replication, suggesting the functional importance of the mature 6K1. We further found that green fluorescent protein-tagged 6K1 formed punctate inclusions at the viral early infection stage and colocalized with chloroplast-bound viral replicase elements 6K2 and NIb. Taken together, our results suggest that 6K1 is required for viral replication and is an important viral element of the viral replication complex at the early infection stage. IMPORTANCE: Potyviruses account for more than 30% of known plant viruses and consist of many agriculturally important viruses. The genomes of potyviruses encode two polyproteins that are proteolytically processed into 11 mature proteins, with the majority of them having been at least partially functionally characterized. However, the functional role of a small protein named 6K1 remains obscure. In this study, we showed that deletion of 6K1 or a short motif/region of 6K1 in the full-length cDNA clones of plum pox virus abolishes viral replication and that mutation of the N- or C-terminal cleavage sites of 6K1 to prevent its release from the polyprotein greatly attenuates or completely inhibits viral replication, suggesting its important role in potyviral infection. We report that 6K1 forms punctate structures and targets the replication vesicles in PPV-infected plant leaf cells at the early infection stage. Our data reveal that 6K1 is an important viral protein of the potyviral replication complex.


Assuntos
Doenças das Plantas/virologia , Vírus Eruptivo da Ameixa/genética , Vírus Eruptivo da Ameixa/fisiologia , Prunus persica/virologia , Tabaco/virologia , Proteínas Virais/metabolismo , Replicação Viral , Canadá , Proteínas de Fluorescência Verde , Folhas de Planta/virologia , Vírus Eruptivo da Ameixa/química , Poliproteínas/genética , Processamento de Proteína Pós-Traducional , Proteólise , Deleção de Sequência , Proteínas Virais/química , Proteínas Virais/genética , Proteínas Virais/isolamento & purificação
15.
Curr Opin Virol ; 17: 19-24, 2016 04.
Artigo em Inglês | MEDLINE | ID: mdl-26651024

RESUMO

As an organelle only found in plant cells and some protists, the chloroplast is not only the main metabolic energy originator, but also the abiotic/biotic stress sensor and defense signal generator. For a long time, chloroplasts have been recognized as a common target by many plant viruses. Viruses may directly modify chloroplast membranes to assemble their replication complex for viral genome replication. Viruses may downregulate chloroplast-related and photosynthesis-related genes via an as yet unknown mechanism to support their infection. Viruses may also interrupt functionality of the photosynthetic machinery through protein-protein interactions. This review briefly summarizes current knowledge about modifications of the photosynthetic machinery by plant viruses, highlights the important role of chloroplasts in the infection process and discusses chloroplast-associated pathogenesis.


Assuntos
Cloroplastos/metabolismo , Cloroplastos/virologia , Vírus de Plantas/patogenicidade , Replicação Viral , Cloroplastos/genética , Cloroplastos/ultraestrutura , Metaboloma , Modelos Biológicos , Fotossíntese , Células Vegetais/metabolismo , Doenças das Plantas/virologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Transcriptoma , Proteínas Virais/metabolismo
16.
J Zhejiang Univ Sci B ; 16(11): 904-13, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26537208

RESUMO

OBJECTIVE: Injury and deficiency of the lacrimal duct epithelium (LDE) can lead to a variety of lacrimal diseases. The purpose of this study was to characterize potential candidate cells for constructing a tissue-engineered LDE. METHODS: Different areas of the conjunctiva and lacrimal duct tissue were removed from male adult New Zealand white rabbits for histological evaluation. Hematoxylin and eosin staining and immunohistochemical staining of cytokeratin AE1+AE3, cytokeratin 4, Ki-67, and MUC5AC were observed by light microscopy. The surface morphologies of different epithelial tissues and cellular structures were examined using field-emission scanning electron microscopy and transmission electron microscopy. Epithelial cells were isolated from tissues and identified by specific markers. In vitro, proliferative ability and Western blot analyses of the proliferating cell nuclear antigen (PCNA) of different epithelial cells cultured in identical environments were investigated and compared. RESULTS: Histologically, the epithelial specific markers, cytokeratin AE1+AE3 and cytokeratin 4, were expressed in the conjunctiva epithelium and the LDE. Notably, highly proliferative cells stained with Ki-67 were concentrated under the epithelium in a dome structure of the posterior palpebral conjunctiva. Differentiated goblet cells were also found to a lesser extent in this region. Primary palpebral and fornical conjunctival epithelial cells (PFCECs), bulbar conjunctival epithelial cells (BCECs), and lacrimal duct epithelial cells (LDECs) were successfully separated from tissues. In vitro, rabbit PFCECs and LDECs grew faster and expressed more PCNA than BCECs. CONCLUSIONS: PFCECs are anatomically similar to LDECs. They also have similar morphological characteristics, immune phenotypes, and proliferation features. PFCECs are therefore potential candidate cells to replace LDECs in tissue engineering to treat lacrimal duct diseases.


Assuntos
Células Epiteliais/citologia , Aparelho Lacrimal/citologia , Engenharia Tecidual , Animais , Proliferação de Células , Células Cultivadas , Células Epiteliais/ultraestrutura , Imuno-Histoquímica , Aparelho Lacrimal/ultraestrutura , Masculino , Microscopia Eletroquímica de Varredura , Coelhos
17.
Virology ; 485: 439-51, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26351203

RESUMO

Double-stranded RNA (dsRNA) is an important type of RNA that plays essential roles in diverse cellular processes in eukaryotic organisms and a hallmark in infections by positive-sense RNA viruses. Currently, no in vivo technology has been developed for visualizing dsRNA in living cells. Here, we report a dsRNA binding-dependent fluorescence complementation (dRBFC) assay that can be used to efficiently monitor dsRNA distribution and dynamics in vivo. The system consists of two dsRNA-binding proteins, which are fused to the N- and C-terminal halves of the yellow fluorescent protein (YFP). Binding of the two fusion proteins to a common dsRNA brings the split YFP halves in close proximity, leading to the reconstitution of the fluorescence-competent structure and restoration of fluorescence. Using this technique, we were able to visualize the distribution and trafficking of the replicative RNA intermediates of positive-sense RNA viruses in living cells.


Assuntos
Proteínas Luminescentes/metabolismo , RNA de Cadeia Dupla/metabolismo , Proteínas de Ligação a RNA/metabolismo , Rastreamento de Células/métodos , Espaço Intracelular/metabolismo , Microscopia Confocal , Modelos Biológicos , Ligação Proteica , Transporte Proteico , RNA Viral/genética , RNA Viral/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo
18.
Onco Targets Ther ; 8: 1017-24, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25999735

RESUMO

PURPOSE: Primary ductal adenocarcinoma (PDA) of the lacrimal gland is a rare malignant epithelial tumor, and its clinicopathological characteristics are still unclear. This study aimed to report a novel case of PDA of the lacrimal gland in the People's Republic of China, as well as to determine its histopathological and immunohistochemical characteristics to support early diagnosis and direct further therapy. PATIENT AND METHODS: Clinical data (including ocular examination, computed tomography, magnetic resonance imaging, positron emission tomography, mammography, and serum tumor marker examination) and treatment of a 49-year-old woman with a left lacrimal gland mass, which was diagnosed as PDA of the lacrimal gland, were reported. Histopathological and immunohistochemical studies were performed. Eleven papers regarding this uncommon neoplasm were reviewed. RESULTS: Histopathologically, most of the tumor cells featured abundant granular eosinophilic cytoplasm, while few of them had a frothy appearance. The mass showed breast ductal carcinoma-like structural features, which most commonly demonstrated central necrosis, while less of these features showed cord-like infiltration. Immunohistochemically, the tumor cells were positive for GCDFP-15, CK 18 (++), AR (90%), Her-2 (+++), P53 (100%), and Ki-67 (with a proliferation index approximately 60%), while they were negative for ER, PR, P63, calponin, and CD 117. CONCLUSION: This was the first Chinese case of PDA of the lacrimal gland that had been reported. We suggested that GCDFP-15, AR, and Her-2 should be tested as biomarkers for ductal adenocarcinoma of lacrimal gland to confirm diagnosis, guide therapy, and further predict prognosis.

19.
Int J Clin Exp Med ; 8(10): 19524-9, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26770604

RESUMO

Ocular sebaceous adenocarcinoma is a rare malignant tumor. We report an unusual case of orbital sebaceous adenocarcinoma in a 48-year-old woman. The patient underwent F-18 2-fluoro-2-deoxy-glucose positron emission tomography/computed tomography ((18)F-FDG-PET/CT) to determine the initial clinical stage. (18)F-FDG-PET/CT showed strong uptake in the orbital lesion, but neither regional lymphatic nor distant metastases were seen. (18)F-FDG-PET/CT plays a role in the diagnosis, staging, restaging, and follow-up of ocular sebaceous carcinoma, although its application has some limitations in other primary and ocular malignancies due to the rarity of ocular sebaceous carcinoma.

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