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[PMID]: 25592246
[Au] Autor:Muñoz-Gutiérrez JF; Schneider DA; Baszler TV; Greenlee JJ; Nicholson EM; Stanton JB
[Ad] Address:Department of Microbiology and Pathology, College of Veterinary Medicine, Washington State University, PO Box 64700, Pullman, WA 99164-7010, United States. Electronic address: jmunoz@vetmed.wsu.edu....
[Ti] Title:hTERT-immortalized ovine microglia propagate natural scrapie isolates.
[So] Source:Virus Res;198:35-43, 2015 Feb 16.
[Is] ISSN:1872-7492
[Cp] Country of publication:Netherlands
[La] Language:eng
[Ab] Abstract:Ex vivo propagation of natural prion isolates (i.e., propagated solely in the natural host) is crucial for the characterization and study of transmissible spongiform encephalopathies (TSEs). Several well-established, prion-permissive cell culture systems are available; however, only a few cell lines are permissive to natural prion isolates and these cells are not pathophysiologically relevant (e.g., renal epithelium and fibroblast-like cells). Therefore, a pathophysiologically relevant cell line derived from a natural TSE host could be used for propagation of natural prion isolates. In this study, ovine brain macrophages (microglia) were immortalized by transfection with the human telomerase reverse transcriptase (hTERT) gene to identify cell lines (hTERT-microglia) permissive to natural scrapie prion isolates. Following transfection, hTERT-microglia were passaged up to 100 times and their lifespan was significantly longer compared to parental cells (Fisher's exact test, P<0.001). Multiple sublines were permissive to cell culture-adapted prions; two sublines were also permissive to natural scrapie isolates (i.e., derived from brain homogenates of sheep infected with scrapie). Prion infectivity and partial protease resistance of the prion protein were maintained in hTERT-microglia. Comparisons between scrapie-permissive and non-permissive hTERT-microglia sublines revealed that overall quantity of the normal cellular prion protein was not associated with prion permissiveness. The use of hTERT-microglia in future TSE studies may be more germane to the characterization of the cellular and subcellular pathophysiology of natural scrapie prion isolates and to investigate host-specific factors involved in prion replication.
[Pt] Publication type:JOURNAL ARTICLE
[Em] Entry month:1502
[Js] Journal subset:IM
[St] Status:In-Data-Review

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[PMID]: 25281761
[Au] Autor:Kleppa E; Holmen SD; Lillebø K; Kjetland EF; Gundersen SG; Taylor M; Moodley P; Onsrud M
[Ad] Address:Norwegian Centre for Imported and Tropical Diseases, Department of Infectious Diseases, Oslo University Hospital Ullevaal, Oslo, Norway Faculty of Medicine, University of Oslo, Oslo, Norway....
[Ti] Title:Cervical ectopy: associations with sexually transmitted infections and HIV. A cross-sectional study of high school students in rural South Africa.
[So] Source:Sex Transm Infect;91(2):124-9, 2015 Mar.
[Is] ISSN:1472-3263
[Cp] Country of publication:England
[La] Language:eng
[Ab] Abstract:OBJECTIVES: It has been hypothesised that ectopy may be associated with increased susceptibility to sexually transmitted infections (STIs). In this cross-sectional study, we wanted to explore the association between STIs (including HIV) and cervical ectopy. METHODS: We included 700 sexually active young women attending randomly selected high schools in a rural district in KwaZulu-Natal, South Africa. The district is endemic of HIV and has a high prevalence of STIs. We did computer-assisted measurements of the ectocervical area covered by columnar epithelium (ectopy) in colposcopic images and STI analyses on cervicovaginal lavage and serum samples. All participating women answered a questionnaire about sexual behaviour and use of contraceptives. RESULTS: The mean age was 19.1 years. Ectopy was found in 27.2%, HIV in 27.8%, chlamydia in 25.3% and gonorrhoea in 15.6%. We found that age, parity, chlamydia and gonorrhoea, years since menarche, years since sexual debut and number of sexual partners were associated with ectopy. In multivariate analysis with chlamydia infection as the dependent variable, women with ectopy had increased odds of having chlamydia infection (adjusted OR 1.78, p=0.033). In women under 19 years of age, we found twofold higher odds of being HIV-positive for those with ectopy (OR 2.19, p=0.014). CONCLUSIONS: In conclusion, cervical ectopy is associated with Chlamydia trachomatis infection and HIV in the youngest women.
[Pt] Publication type:JOURNAL ARTICLE
[Em] Entry month:1502
[Js] Journal subset:IM
[St] Status:In-Data-Review
[do] DOI:10.1136/sextrans-2014-051674

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[PMID]: 25428941
[Au] Autor:Isik AÜ; Arslan S; Arslan E; Baykal S
[Ad] Address:M.D., Department of Otorhinolaryngology, School of Medicine, Karadeniz Technical University, Turkey....
[Ti] Title:A giant frontoethmoid mucocele with intracranial extension.
[So] Source:Scott Med J;60(1):e1-3, 2015 Feb.
[Is] ISSN:0036-9330
[Cp] Country of publication:Scotland
[La] Language:eng
[Ab] Abstract:Mucoceles are mucus-containing cysts lined by epithelium. Although benign, they may show expansive growth and remain undiagnosed until symptoms due to compression of surrounding structures arise. We report a rare case of frontoethmoid mucocele with intracranial extension in an 80-year-old woman with complaints of headache, right diplopia and proptosis. A right frontoorbital craniotomy was performed, and a mucocele in the frontal sinus extending into the frontal lobe and orbit was totally removed. The patient was successfully treated without any complication. The two-year follow-up results were satisfactory. Magnetic resonance imaging excluded any recurrence of the mucocele. Combined intranasal and transcranial approach is necessary to treat giant frontoetmoid mucoceles with intracranial extension.
[Pt] Publication type:JOURNAL ARTICLE
[Em] Entry month:1502
[Js] Journal subset:IM
[St] Status:In-Data-Review
[do] DOI:10.1177/0036933014561949

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[PMID]: 25008036
[Au] Autor:Doumas S; Paterson JC; Norris PM; Tighe JV; Newman L; Bisase BS; Kolokotronis AE; Barrett AW
[Ad] Address:Queen Victoria Hospital NHSF Trust, East Grinstead, West Sussex, UK.
[Ti] Title:Fractalkine (CX3CL1) and fractalkine receptor (CX3CR1) in squamous cell carcinoma of the tongue: Markers of nerve invasion?
[So] Source:Oral Maxillofac Surg;19(1):61-4, 2015 Mar.
[Is] ISSN:1865-1569
[Cp] Country of publication:Germany
[La] Language:eng
[Ab] Abstract:PURPOSE: Perineural invasion (PNI) in oral squamous cell carcinoma (SCC) is an independent predictor of poor prognosis. As PNI is not always identified with routine histology, a surrogate marker of PNI would improve detection and better inform treatment planning. The chemokines fractalkine (CX3CL1) and its receptor (CX3CR1) have shown such potential in other cancers, but have yet to be investigated with respect to PNI in oral SCC. METHODS: Thirty SCCs of the tongue in which PNI was identified histologically, and 30 in which it was not, were stained for fractalkine and fractalkine receptor using polyclonal antibodies and an immunoperoxidase technique. Tumours were assessed as either positive or negative; no attempt was made to subjectively assess staining intensity or extent. RESULTS: Both markers labelled myofibroblasts in the stroma surrounding the tumour, various neural components, leucocytes, endothelium and salivary myoepithelial cells. Fractalkine also labelled salivary ductal epithelium, vascular smooth muscle and 12/30 SCC which showed PNI. Eight of 30 positive SCCs in which PNI was not identified were also positive for this marker. There was no statistically significant association between fractalkine staining and PNI (p = 0.273). No SCC was positive for fractalkine receptor, but immune dendritic cells within tumour islands were strongly positive, as was striated muscle. CONCLUSIONS: Neither fractalkine nor fractalkine receptor is a reliable surrogate marker of PNI in lingual SCC.
[Pt] Publication type:JOURNAL ARTICLE
[Em] Entry month:1502
[Js] Journal subset:D; IM
[St] Status:In-Data-Review
[do] DOI:10.1007/s10006-014-0455-4

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[PMID]: 25695148
[Au] Autor:Kuwahara A; Ozone C; Nakano T; Saito K; Eiraku M; Sasai Y
[Ad] Address:1] Neurogenesis and Organogenesis Group, RIKEN Center for Developmental Biology, 2-2-3 Manatojima-Minamimachi, Chuo, Kobe 650-0047, Japan [2] Human Stem Cell Technology Unit, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo, Kobe 650-0047, Japan [3] Environmental Health Sci...
[Ti] Title:Generation of a ciliary margin-like stem cell niche from self-organizing human retinal tissue.
[So] Source:Nat Commun;6:6286, 2015.
[Is] ISSN:2041-1723
[Cp] Country of publication:England
[La] Language:eng
[Ab] Abstract:In the developing neural retina (NR), multipotent stem cells within the ciliary margin (CM) contribute to de novo retinal tissue growth. We recently reported the ability of human embryonic stem cells (hESCs) to self-organize stratified NR using a three-dimensional culture technique. Here we report the emergence of CM-like stem cell niches within human retinal tissue. First, we developed a culture method for selective NR differentiation by timed BMP4 treatment. We then found that inhibiting GSK3 and FGFR induced the transition from NR tissue to retinal pigment epithelium (RPE), and that removing this inhibition facilitated the reversion of this RPE-like tissue back to the NR fate. This step-wise induction-reversal method generated tissue aggregates with RPE at the margin of central-peripherally polarized NR. We demonstrate that the NR-RPE boundary tissue further self-organizes a niche for CM stem cells that functions to expand the NR peripherally by de novo progenitor generation.
[Pt] Publication type:JOURNAL ARTICLE
[Em] Entry month:1502
[Js] Journal subset:IM
[St] Status:In-Data-Review
[do] DOI:10.1038/ncomms7286

  6 / 254111 MEDLINE  
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[PMID]: 25673096
[Au] Autor:Murali K; Kenesei K; Li Y; Demeter K; Környei Z; Madarász E
[Ad] Address:Laboratory of Cellular and Developmental Neurobiology, Institute of Experimental Medicine of Hungarian Academy of Sciences, 43 Szigony u., H-1083-Budapest, Hungary. murali.kumarasamy@koki.mta.hu.
[Ti] Title:Uptake and bio-reactivity of polystyrene nanoparticles is affected by surface modifications, ageing and LPS adsorption: in vitro studies on neural tissue cells.
[So] Source:Nanoscale;7(9):4199-210, 2015 Feb 19.
[Is] ISSN:2040-3372
[Cp] Country of publication:England
[La] Language:eng
[Ab] Abstract:Because of their capacity of crossing an intact blood-brain barrier and reaching the brain through an injured barrier or via the nasal epithelium, nanoparticles have been considered as vehicles to deliver drugs and as contrast materials for brain imaging. The potential neurotoxicity of nanoparticles, however, is not fully explored. Using particles with a biologically inert polystyrene core material, we investigated the role of the chemical composition of particle surfaces in the in vitro interaction with different neural cell types. PS NPs within a size-range of 45-70 nm influenced the metabolic activity of cells depending on the cell-type, but caused toxicity only at extremely high particle concentrations. Neurons did not internalize particles, while microglial cells ingested a large amount of carboxylated but almost no PEGylated NPs. PEGylation reduced the protein adsorption, toxicity and cellular uptake of NPs. After storage (shelf-life >6 months), the toxicity and cellular uptake of NPs increased. The altered biological activity of "aged" NPs was due to particle aggregation and due to the adsorption of bioactive compounds on NP surfaces. Aggregation by increasing the size and sedimentation velocity of NPs results in increased cell-targeted NP doses. The ready endotoxin adsorption which cannot be prevented by PEG coating, can render the particles toxic. The age-dependent changes in otherwise harmless NPs could be the important sources for variability in the effects of NPs, and could explain the contradictory data obtained with "identical" NPs.
[Pt] Publication type:JOURNAL ARTICLE
[Em] Entry month:1502
[Js] Journal subset:IM
[St] Status:In-Data-Review
[do] DOI:10.1039/c4nr06849a

  7 / 254111 MEDLINE  
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[PMID]: 25213606
[Au] Autor:Mastorakos P; Kambhampati SP; Mishra MK; Wu T; Song E; Hanes J; Kannan RM
[Ad] Address:Center for Nanomedicine at the Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. krangar1@jhmi.edu.
[Ti] Title:Hydroxyl PAMAM dendrimer-based gene vectors for transgene delivery to human retinal pigment epithelial cells.
[So] Source:Nanoscale;7(9):3845-56, 2015 Feb 19.
[Is] ISSN:2040-3372
[Cp] Country of publication:England
[La] Language:eng
[Ab] Abstract:Ocular gene therapy holds promise for the treatment of numerous blinding disorders. Despite the significant progress in the field of viral and non-viral gene delivery to the eye, significant obstacles remain in the way of achieving high-level transgene expression without adverse effects. The retinal pigment epithelium (RPE) is involved in the pathogenesis of retinal diseases and is a key target for a number of gene-based therapeutics. In this study, we addressed the inherent drawbacks of non-viral gene vectors and combined different approaches to design an efficient and safe dendrimer-based gene-delivery platform for delivery to human RPE cells. We used hydroxyl-terminated polyamidoamine (PAMAM) dendrimers functionalized with various amounts of amine groups to achieve effective plasmid compaction. We further used triamcinolone acetonide (TA) as a nuclear localization enhancer for the dendrimer-gene complex and achieved significant improvement in cell uptake and transfection of hard-to-transfect human RPE cells. To improve colloidal stability, we further shielded the gene vector surface through incorporation of PEGylated dendrimer along with dendrimer-TA for DNA complexation. The resultant complexes showed improved stability while minimally affecting transgene delivery, thus improving the translational relevance of this platform.
[Pt] Publication type:JOURNAL ARTICLE
[Em] Entry month:1502
[Js] Journal subset:IM
[St] Status:In-Data-Review
[do] DOI:10.1039/c4nr04284k

  8 / 254111 MEDLINE  
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[PMID]: 25697537
[Au] Autor:Rossmiller BP; Ryals RC; Lewin AS
[Ad] Address:Department of Opthalmology, University of Florida, Box 100284, Gainesville, FL, 32610-0284, USA.
[Ti] Title:Gene therapy to rescue retinal degeneration caused by mutations in rhodopsin.
[So] Source:Methods Mol Biol;1271:391-410, 2015.
[Is] ISSN:1940-6029
[Cp] Country of publication:United States
[La] Language:eng
[Ab] Abstract:Retinal gene therapy has proven safe and at least partially successful in clinical trials and in numerous animal models. Gene therapy requires characterization of the progression of the disease and understanding of its genetic cause. Testing gene therapies usually requires an animal model that recapitulates the key features of the human disease, though photoreceptors and cells of the retinal pigment epithelium produced from patient-derived stem cells may provide an alternative test system for retinal gene therapy. Gene therapy also requires a delivery system that introduces the therapeutic gene to the correct cell type and does not cause unintended damage to the tissue. Current systems being tested in the eye are nanoparticles, pseudotyped lentiviruses, and adeno-associated virus (AAV) of various serotypes. Here, we describe the techniques of AAV vector design as well as the in vivo and ex vivo tests necessary for assessing the efficacy of retinal gene therapy to treat retinal degeneration caused by mutations in the rhodopsin gene.
[Pt] Publication type:JOURNAL ARTICLE
[Em] Entry month:1502
[Js] Journal subset:IM
[St] Status:In-Data-Review
[do] DOI:10.1007/978-1-4939-2330-4_25

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[PMID]: 25697534
[Au] Autor:Perusek L; Maeda A; Maeda T
[Ad] Address:Department of Ophthalmology and Visual Sciences, School of Medicine, Case Western Reserve University, Adelbert Road 2085, Cleveland, OH, 44106, USA.
[Ti] Title:Supplementation with vitamin a derivatives to rescue vision in animal models of degenerative retinal diseases.
[So] Source:Methods Mol Biol;1271:345-62, 2015.
[Is] ISSN:1940-6029
[Cp] Country of publication:United States
[La] Language:eng
[Ab] Abstract:The perception of light begins when photons reach retinal tissue located at the back of the eye and photoisomerize the visual chromophore 11-cis-retinal to all-trans-retinal within photoreceptor cells. Isomerization of 11-cis-retinal activates the protein rhodopsin located in photoreceptor outer segments, thereby inducing a phototransduction cascade leading to visual perception. To maintain vision, 11-cis-retinal is regenerated in the retinal pigmented epithelium (RPE) via the visual cycle and delivered back to the photoreceptor cells where it may again bind to rhodopsin. Distinct pathological mechanisms have been observed to contribute to inherited retinal degenerative diseases including severe delay in 11-cis-retinal regeneration and delayed clearance of all-trans-retinal, which leads to the accumulation of harmful retinoid by-products. In the last decade, our group has conducted several proof-of-concept (POC) studies with retinoid derivatives aimed at developing treatments for retinal degenerative diseases caused by an impaired visual cycle. Here, we will introduce experimental procedures, which have been developed for POC studies involving retinoid biology.
[Pt] Publication type:JOURNAL ARTICLE
[Em] Entry month:1502
[Js] Journal subset:IM
[St] Status:In-Data-Review
[do] DOI:10.1007/978-1-4939-2330-4_22

  10 / 254111 MEDLINE  
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[PMID]: 25697533
[Au] Autor:Adler L; Boyer NP; Chen C; Koutalos Y
[Ad] Address:Departments of Ophthalmology and Neurosciences, Medical University of South Carolina, 167 Ashley Ave., Charleston, SC, 29425, USA.
[Ti] Title:Kinetics of Rhodopsin's Chromophore Monitored in a Single Photoreceptor.
[So] Source:Methods Mol Biol;1271:327-43, 2015.
[Is] ISSN:1940-6029
[Cp] Country of publication:United States
[La] Language:eng
[Ab] Abstract:Absorption of light isomerizes the retinyl chromophore of the photoreceptor pigment rhodopsin from 11-cis to all-trans, generating the photoactivated rhodopsin form. The photoisomerization of the chromophore however destroys rhodopsin, and its regeneration requires the removal of the all-trans and the supply of fresh 11-cis chromophore. The all-trans chromophore is removed through a series of steps beginning with its release from photoactivated rhodopsin in the form of all-trans-retinal, leaving behind the apoprotein opsin. All-trans-retinal is then reduced to all-trans-retinol, which is transported out of the photoreceptor. Rhodopsin is regenerated from opsin and fresh 11-cis-retinal arriving to the photoreceptor from the retinal pigment epithelium. Both all-trans and 11-cis-retinal can form precursors of lipofuscin, a pigment that accumulates with age in the lysosomal compartment of the retinal pigment epithelium. All-trans-retinal, all-trans-retinol, and lipofuscin precursors all emit significant and distinct fluorescence signals, allowing their monitoring in single photoreceptor cells with fluorescence imaging. Here we describe the procedures for measuring these fluorophores in single mouse rod photoreceptors.
[Pt] Publication type:JOURNAL ARTICLE
[Em] Entry month:1502
[Js] Journal subset:IM
[St] Status:In-Data-Review
[do] DOI:10.1007/978-1-4939-2330-4_21


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