Late-onset cytomegalovirus (CMV) disease commonly occurs after discontinuation of antiviral prophylaxis. We determined the utility of testing CD8+ T-cell response against CMV as a predictor of late-onset CMV disease after a standard course of antiviral prophylaxis. Transplant patients at high-risk for CMV disease were enrolled. CD8+ T-cell-mediated immunity (CMI) was tested using the QuantiFERON-CMV assay at baseline, 1, 2 and 3 months posttransplant by measurement of interferon-c response to whole blood stimulation with a 21-peptide pool. The primary outcome was the ability of CMI testing to predict CMV disease in the first 6 months posttransplant. There were 108 evaluable patients (D+/R+ n = 39; D-/R+ n = 34; D+/R-n = 35) of whom 18 (16.7%) developed symptomatic CMV disease. At the end of prophylaxis, CMI was detectable in 38/108 (35.2%) patients (cutoff 0.1 IU/mL interferonc ). CMV disease occurred in 2/38 (5.3%) patients with a detectable interferon-c response versus 16/70 (22.9%) patients with a negative response; p = 0.038. In the subgroup of D+/R-patients, CMV disease occurred in 1/10 (10.0%) patients with a detectable interferon-c response (cutoff 0.1 IU/mL) versus 10/25 (40.0%) patients with a negative CMI, p = 0.12. Monitoring of CMI may be useful for predicting late-onset CMV disease.

The authors note, "Figs. 2, 3, and 5 were not in compliance with PNAS figure preparation policies because some gel lanes from noncontiguous experiments were spliced together. The corrected figures shown below have white spaces inserted to indicate noncontiguous experiments." Fig. 2. In vitro transcription/translation and immunoprecipitation of translated products using antiserum raised against the scRYMV ORF-encoded protein.(A) Lane 2 shows 19-kDa and 16-kDa proteins from reaction using pET52 DNA, whereas lane 3 shows only the 19 kDa from reaction using the truncated pETdimer DNA. Lane 1: Negative control reaction using pET29 (empty vector). Arrows on left indicate molecular size of proteins, from top to bottom: 36 kDa, 19 kDa, and 16 kDa. (B) In vitro translation reaction depicting the 16-kDa protein from the scRYMV circular RNA (purified from denaturing polyacrylamide gels) is shown in lane 3, whereas that of the reaction from total RNA of RYMV-infected rice is shown in lane 2. Lane 5 demonstrates the 16-kDa product from reaction using total viral RNA extracted from RYMV virus particles, and lane 4 shows the enhanced 16-kDa signal from reaction using RYMV total viral RNA but supplemented with the same amount of gel-purified circular RNA as that used in the reaction of lane 3. Lanes 1 and 6 represent negative control reactions using healthy rice and the endogenous empty lysate, respectively. Arrows on right depict the position of molecular weight markers 25, 16, and 14 kDa from top to bottom, respectively. (C) Northern analysis to detect the nature of scRYMV RNA species. Denaturing 4-20% PAGE in presence of 8 M urea was carried out. Lanes 2 and 3 demonstrate presence of linear (marked as "L"), circular ("C"), dimer ("D"), and trimer ("T") forms of the scRYMV RNA in total RNA preparations from RYMV-infected rice and RYMV particles, respectively. Lane 1 shows the negative control of total RNA from healthy rice. Lane 4: 7 M urea-PAGE stained with ethidium bromide and showing the purified circular RNA extracted from the band corresponding to circular RNA (shown in lane 3). This purified circular RNA is used for in vitro translation. Arrows indicate the positions of RNA size markers: 220 (marked as "L" or "C"), 440 ("D"), and 660 ("T") nt.

Glaucoma is characterized by loss of retinal nerve fibers, structural changes to the optic nerve, and an associated change in visual function. The major risk factor for glaucoma is an increase in intraocular pressure (IOP). However, it has been demonstrated that a subset of glaucoma patients exhibit optic neuropathy despite a normal range of IOP. It has been proposed that primary open angle glaucoma could be associated with structural abnormalities and/or functional dysregulation of the vasculature supplying the optic nerve and surrounding retinal tissue. Under normal conditions, blood flow is autoregulated, i.e., maintained at a relatively constant level, in the retina and ONH, irrespective of variation in ocular perfusion pressure. A number of factors released by the vascular endothelium, including endothelin-1 and nitric oxide, are suggested to play an important role in the regulation of local perfusion in the retina and ONH. Most work to-date has investigated homeostatic hemodynamic parameters in glaucoma, rather than the measurement of the hemodynamic response to a provocation. Future work should comprehensively assess blood flow in all the ocular vascular beds and blood vessels supplying the eye in response to standardized stimuli in order to better understand the pathophysiology of glaucomatous optic neuropathy.

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