It is a remarkable feature for a retrovirus that an infection with feline leukemia virus (FeLV) can result in various outcomes. Whereas some cats contain the infection and show a regressive course, others stay viremic and succumb to the infection within a few years. We hypothesized, that differences in the infection outcome might be causally linked to the viral RNA and provirus loads within the host and these loads therefore may give additional insight into the pathogenesis of the virus. Thus, the goals of the present study were to follow-up on experimentally infected cats and investigate tissues from cats with different infection outcomes using sensitive, specific TaqMan real-time PCR and reverse transcriptase (RT)-PCR. Nineteen experimentally FeLV-A/Glasgow-1-infected cats were categorized into having regressive, progressive or reactivated FeLV infection according to follow-up of FeLV p27 antigen detection in the blood. Remarkably, regressively infected cats showed detectable provirus and viral RNA loads in almost all of the 27 tested tissues, even many years after virus exposure. Moreover, some regressively infected cats reactivated the infection, and these cats had intermediate to high viral RNA and provirus tissue loads. The highest loads were found in viremic cats, independent of their health status. Tissues that represented sites of virus replication and shedding revealed the highest viral RNA and provirus loads, while the lowest loads were present in muscle and nerve tissues. A supplementary analysis of 20 experimentally infected cats with progressive infection revealed a median survival time of 3.1 years (range from 0.6 to 6.5 years); 70% (n=14) of these cats developed lymphoma, while leukemia and nonregenerative anemia were observed less frequently. Our results demonstrate that the different infection outcomes are associated with differences in viral RNA and provirus tissue loads. Remarkably, no complete clearance of FeLV viral RNA or provirus was detected in cats with regressive infection, even up to 12 years after exposure. In several cases FeLV reactivation could be observed. Thus, retroviruses integrated as provirus into the host's genome, could not be eliminated completely by the host and maintained their full potential for replication and reactivation. DOI: https://doi.org/10. 1016/j.virusres.2014.12.025 Posted at the Zurich Open Repository and Archive, University of Zurich ZORA URL: https://doi.org/10.5167/uzh-105626 Accepted Version Originally published at: Helfer-Hungerbuehler, A Katrin; Widmer, Stefan; Kessler, Yvonne; Riond, Barbara; Boretti, Felicitas S; Grest, Paula; Lutz, Hans; Hofmann-Lehmann, Regina (2015). Long-term follow up of feline leukemia virus infection and characterization of viral RNA loads using molecular methods in tissues of cats with different infection outcomes. Virus Research,
Quantitative real-time PCR (qPCR) is broadly used to detect and quantify nucleic acid targets. In order to determine cell copy number and genome equivalents, a suitable reference gene that is present in a defined number in the genome is needed, preferably as a single copy gene. For most organisms, a variable number of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) pseudogenes have been reported. However, it has been suggested that a single-copy of the GAPDH pseudogene is present in the feline genome and that a GAPDH assay can therefore be used to quantify feline genomic DNA (gDNA). The aim of this study was to determine whether one or more GAPDH pseudogenes are present in the feline genome and to provide a suitable alternative qPCR system for the quantification of feline cell copy number and genome equivalents. Bioinformatics and sequencing results revealed that not just one but several closely related GAPDH-like sequences were present in the cat genome. We thus identified, developed, optimized, and validated an alternative reference gene assay using feline albumin (fALB). Our data emphasize the need for an alternative reference gene, apart from the GAPDH pseudogene, for the normalization of gDNA levels. We recommend using the fALB qPCR assay for future studies.
The currently accepted intravitreal use of amikacin may cause retinal toxicity.
LMG is a rare but highly malignant Epstein-Barr virus associated NK/T-cell lymphoma that can occur in healthy, immune competent Caucasians. This is the first reported case of an LMG in an immune-competent Caucasian patient with primary ocular manifestation. The LMG has a high mortality rate despite systemic treatment and can be lethal within a few months or even weeks.
Cognitive abilities such as attention or working memory can support older adults during speech perception. However, cognitive abilities as well as speech perception decline with age, leading to the expenditure of effort during speech processing. This longitudinal study therefore investigated age-related differences in electrophysiological processes during speech discrimination and assessed the extent of enhancement to such cognitive auditory processes through repeated auditory exposure. For that purpose, accuracy and reaction time were compared between 13 older adults (62-76 years) and 15 middle-aged (28-52 years) controls in an active oddball paradigm which was administered at three consecutive measurement time points at an interval of 2 wk, while EEG was recorded. As a standard stimulus, the nonsense syllable / 0 a:Sa/was used, while the nonsense syllable / 0 a:sa/ and a morphing between / 0 a:Sa/ and / 0 a:sa/ served as deviants. N2b and P3b ERP responses were evaluated as a function of age, deviant, and measurement time point using a data-driven topographical microstate analysis. From middle age to old age, age-related decline in attentive perception (as reflected in the N2b-related microstates) and in memory updating and attentional processes (as reflected in the P3b-related microstates) was found, as indicated by both lower neural responses and later onsets of the respective cortical networks, and in age-related changes in frontal activation during attentional stimulus processing. Importantly, N2b-and P3b-related microstates changed as a function of repeated stimulus exposure in both groups. This research therefore suggests that experience with auditory stimuli can support auditory neurocognitive processes in normal hearing adults into advanced age.
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