Cells that actively transcribe HIV-1 have been defined as the “active viral reservoir” in HIV-infected individuals. However, important technical limitations have precluded the characterization of this specific viral reservoir during both treated and untreated HIV-1 infections. Here, we used a novel single-cell RNA fluorescence in situ hybridization-flow cytometry (FISH-flow) assay that requires only 15 million unfractionated peripheral blood mononuclear cells (PBMCs) to characterize the specific cell subpopulations that transcribe HIV RNA in different subsets of CD4+ T cells. In samples from treated and untreated HIV-infected patients, effector memory CD4+ T cells were the main cell population supporting HIV RNA transcription. The number of cells expressing HIV correlated with the plasma viral load, intracellular HIV RNA, and proviral DNA quantified by conventional methods and inversely correlated with the CD4+ T cell count and the CD4/CD8 ratio. We also found that after ex vivo infection of unstimulated PBMCs, HIV-infected T cells upregulated the expression of CD32. In addition, this new methodology detected increased numbers of primary cells expressing viral transcripts and proteins after ex vivo viral reactivation with latency reversal agents. This RNA FISH-flow technique allows the identification of the specific cell subpopulations that support viral transcription in HIV-1-infected individuals and has the potential to provide important information on the mechanisms of viral pathogenesis, HIV persistence, and viral reactivation.
The medical records of 468 horses that underwent 490 exploratory laparotomies for the correction of gastrointestinal diseases were reviewed to search for differences between Andalusian horses and other breeds. The seasonal distribution of surgical colics and their outcome and complications were also investigated. Bivariant analysis was used to compare the horses' age, gender and breed with the type of surgery, the bowel affected and the type of colic, and all these variables were compared in relation to euthanasia during surgery, complications, short-term survival and seasonal distribution. A total of 405 horses survived the surgery and 329 were discharged from the hospital. Horses less than one year old had better short-term survival than older horses. Andalusian horses suffered more inguinal hernias than the other breeds and were more prone to suffer laminitis as a complication. Colic surgery and inguinal hernias were also more common in the summer.
Acute syphilitic posterior placoid chorioretinopathy (ASPPC) is one of the rarest ocular manifestations of syphilis. The pathophysiology of this entity is still unknown. We report the outer retinal findings on en face optical coherence tomography (OCT) and the alteration of choriocapillaris flow findings on OCT angiography in a patient with ASPPC at the time of presentation, after penicillin treatment completion and during follow-up.
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