Journal: Emerging Infectious Diseases

Article Title: Streptococcus gallolyticus and Bacterial Endocarditis in Swine, United States, 2015–2020

doi: 10.3201/eid2801.210998

Figure Lengend Snippet: Lesions associated with swine vegetative endocarditis, United States, 2015–2020. A) Macroscopic findings of vegetative growth on the left atrioventricular heart valve leaflets. B) Histopathologic findings of inflammation characterized by necrotic leukocytes (N), fibrin (F), mineralization (M), and myriad bacterial colonization (yellow outline) along the surface of the heart valve (hematoxylin and eosin staining); original magnification ×40. Higher magnification image (inset) shows cocci bacteria in clusters and long chains; original magnification ×1,000. C) Streptococcus gallolyticus directly detected (red) on the surface of the heart valve by RNA in situ hybridization with a probe targeting the helix-hairpin helix domain–containing protein, ComEC/Rec2, and DNA pol III subunit delta genes specific to S. gallolyticus ; original magnification ×40. Higher magnification image (inset) shows the bacteria labeled by the in situ hybridization probe; original magnification ×1,000.

Article Snippet: For selected cases from which pure growth of S. gallolyticus was isolated from the heart valves, we performed bacterial biochemical analyses, sequenced 16S ribosomal RNA, and localized S. gallolyticus within lesions via RNA in situ hybridization (ISH) (RNAscope; TriStar Technology LLC, https://tristargroup.us ).

Techniques: Staining, Bacteria, RNA In Situ Hybridization, Labeling, In Situ Hybridization

Journal: Neurobiology of aging

Article Title: Recombinant adeno-associated virus 2/5-mediated gene transfer is reduced in the aged rat midbrain

doi: 10.1016/j.neurobiolaging.2014.07.047

Figure Lengend Snippet: (a-b) Overview and timeline of the three cohorts of rats used in these studies. Young adult (3 months) and aged (20 months) Sprague Dawley and Fischer344 rats were all injected in the substantia nigra (SN) with the rAAV2/5 GFP viral vector and were sacrificed at indicated time points to harvest tissue for various outcome measures. (c) Recombinant adeno-associated virus (rAAV) genomic map of rAAV2/5 expressing GFP used in all experiments.

Article Snippet: Values are displayed as the mean linearized GFP 2 -ΔCT values ± percent change from young adult ΔCT values (Young Adult, n=6; Aged, n=6). (b) Representative images using RNAscope in situ hybridization for GFP in the Sprague Dawley (SD) midbrain 11 days following rAAV2/5 vector injection.

Techniques: Injection, Plasmid Preparation, Recombinant, Virus, Expressing

Journal: Neurobiology of aging

Article Title: Recombinant adeno-associated virus 2/5-mediated gene transfer is reduced in the aged rat midbrain

doi: 10.1016/j.neurobiolaging.2014.07.047

Figure Lengend Snippet: (a) Representative montage of GFP immunoreactivity within individual midbrain sections 11 days following rAAV2/5 GFP injection to either young adult or aged rats. GFP immunoreactive cells are generally observed spanning the entire AP axis in both young adult and aged rats, with aged rats generally exhibiting smaller areas of transgene expression within individual coronal sections. The total volume of transgene expression in the young adult brain was 205,524,000μm3, or 10.47% of the mesencephalon, whereas the total volume of transgene expression in the aged brain was 135,517,000μm3, or 6.62% of the mesencephalon. (b) Stereological estimates of the number of GFP immunoreactive cells per individual mesencephalic coronal section indicate a general trend of fewer GFP immunoreactive cells within each aged brain section as compared to the comparable section in the young adult. (c) Stereological estimates of the number of GFP immunoreactive cells within the rostral, central, and caudal regions of the area displaying transgene expression. Aged rats possess a trend for fewer GFP immunoreactive cells in the rostral and caudal regions with significantly fewer GFP immunoreactive cells in the central region (*p<0.05) than young adult rats. Values are expressed as the mean ± SEM for each group (Young Adult, n=6; Aged, n=5).

Article Snippet: Values are displayed as the mean linearized GFP 2 -ΔCT values ± percent change from young adult ΔCT values (Young Adult, n=6; Aged, n=6). (b) Representative images using RNAscope in situ hybridization for GFP in the Sprague Dawley (SD) midbrain 11 days following rAAV2/5 vector injection.

Techniques: Injection, Expressing

Journal: Neurobiology of aging

Article Title: Recombinant adeno-associated virus 2/5-mediated gene transfer is reduced in the aged rat midbrain

doi: 10.1016/j.neurobiolaging.2014.07.047

Figure Lengend Snippet: (a) Representative image of TH immunoreactive (THir) neurons in the substantia nigra pars compacta (SNpc) of young adult (Young) and aged (Aged) SD brains at 11 days post-injection. TH immunoreactivity appears equal between hemispheres and age groups. Scale bar = 1000 μm. (b) Stereological quantification of THir neurons in young adult and aged SD rats in the rAAV2/5 GFP injected and uninjected SNpc. No significant reductions in tyrosine hydroxylase immunoreactive (THir) neurons are observed in aged (n=5) as compared to young adult (n=6) rats or in the injected as compared to the uninjected hemisphere 11 days following rAAV2/5 GFP injection. (c) Representative samples of GFP immunoreactivity in the Young and Aged SD rat brain 11 days post-injection. GFP immunoreactivity is observed throughout the midbrain ipsilateral to the injection site (left) with dense cellular staining observed at higher magnfications. GFP immunoreactivity in the midbrain of aged rats is visibly less abundant than that of young adult rats. Scale bars = 1000μm in top panels, 100μm in lower panels and 25μm in the high magnification inserts. (d) Stereological quantification of the total number of GFP immunoreactive cells for young adult (n=6) and aged (n=5) rats revealed a statistically significant (*p<0.05) 40% decrease in GFP immunoreactive cells in the aged brain as compared to the young adult brain. (e-j) GFP co-expression within THir SN neurons in young adult (e-g) and aged (h-j). Numerous THir neurons and non-THir cells expressing GFP are apparent in both age groups. Scale bar = 50μm. (k) Quantification of the number of THir neurons expressing GFP in three sections containing a readily identifiable SNpc in young adult and aged rats. No significant differences were observed in the number of GFPir/THir neurons at these levels of the mesencephalon. (l) Quantification of the percentage THir and non-THir cells that colocalize with GFP expression. No significant differences were observed between young adult and aged rats in the percent of THir neurons or the percent of non-THir cells expressing GFP, although both age groups exhibited significantly more GFPir non-THir cells than GFPir/THir neurons (Aged, n=5; Young Adult, n=6). All values are expressed as the mean ± SEM for each group (Aged, n=5; Young Adult, n=6).

Article Snippet: Values are displayed as the mean linearized GFP 2 -ΔCT values ± percent change from young adult ΔCT values (Young Adult, n=6; Aged, n=6). (b) Representative images using RNAscope in situ hybridization for GFP in the Sprague Dawley (SD) midbrain 11 days following rAAV2/5 vector injection.

Techniques: Injection, Staining, Expressing

Journal: Neurobiology of aging

Article Title: Recombinant adeno-associated virus 2/5-mediated gene transfer is reduced in the aged rat midbrain

doi: 10.1016/j.neurobiolaging.2014.07.047

Figure Lengend Snippet: (a) Representative western blot of GFP immunodetection in striatal samples of young adult and aged Sprague Dawley (SD) rats 11 days post-injection and Fischer344 (F344) rats 12 days post-injection. Young adult rats consistently express more GFP protein than aged rats. (b) Quantification of striatal GFP protein levels in young adult (n=6) and aged (n=5) SD rats 11 days post-injection. Aged rats exhibited significantly lower levels of GFP protein (*p<0.05) than young adult rats. (c) Quantification of striatal GFP protein levels in young adult (n=6) and aged (n=6) F344 rats at 12 days post-injection; aged rats display significantly less GFP protein (**p<0.01) than young adult rats. (d) Quantification of GFP protein levels in the striatum of young adult (n=5) and aged (n=5) SD rats at 3 months post-injection. Aged rats exhibit significantly less GFP protein (*p<0.05) than young adult rats. Values are expressed as the mean optical density scores, normalized to tubulin controls ± SEM for each group. rAAV, recombinant adeno-associated virus.

Article Snippet: Values are displayed as the mean linearized GFP 2 -ΔCT values ± percent change from young adult ΔCT values (Young Adult, n=6; Aged, n=6). (b) Representative images using RNAscope in situ hybridization for GFP in the Sprague Dawley (SD) midbrain 11 days following rAAV2/5 vector injection.

Techniques: Western Blot, Immunodetection, Injection, Recombinant, Virus

Journal: Neurobiology of aging

Article Title: Recombinant adeno-associated virus 2/5-mediated gene transfer is reduced in the aged rat midbrain

doi: 10.1016/j.neurobiolaging.2014.07.047

Figure Lengend Snippet: (a) GFP mRNA quantitation using qPCR of the Fischer344 (F344) substantia nigra (SN). GFP mRNA expression in the aged F344 SN was significantly lower (approximately 4-fold less) than expression levels in the young adult SN (***p<0.001) 12 days following rAAV2/5 vector injection. Values are displayed as the mean linearized GFP 2-ΔCT values ± percent change from young adult ΔCT values (Young Adult, n=6; Aged, n=6). (b) Representative images using RNAscope in situ hybridization for GFP in the Sprague Dawley (SD) midbrain 11 days following rAAV2/5 vector injection. GFP mRNA expression in the young adult rat is observed throughout the midbrain ipsilateral to the injection site, with dense staining observed at higher magnifications. Scale bar = 1000μm in top panel of lower magnification images, 200μm in lower panel images and 50μm in lower panel inserts. GFP mRNA expression in the midbrain of aged rats is visually less abundant than that of young adult rats with robust staining observed at higher magnifications.

Article Snippet: Values are displayed as the mean linearized GFP 2 -ΔCT values ± percent change from young adult ΔCT values (Young Adult, n=6; Aged, n=6). (b) Representative images using RNAscope in situ hybridization for GFP in the Sprague Dawley (SD) midbrain 11 days following rAAV2/5 vector injection.

Techniques: Quantitation Assay, Expressing, Plasmid Preparation, Injection, RNAscope, In Situ Hybridization, Staining