Extensive experimental and clinical data show that the ultrasonic image conveys information on the biochemical composition of the atherosclerotic plaque, ie, the relative content of lipids (hypoechoic), fibrous tissue (hyperechoic), and calcific deposits (very echogenic with shadowing). A more dishomogeneous echo structure of the plaque is also more often associated with clinically complicated carotid plaques. To date, however, the assessment of plaque density and homogeneity by transcutaneous B-mode imaging remains subjective and qualitative. The aim of this study was to assess whether plaque echodensity and homogeneity might be established on a more objective and quantitative basis by description of the spatial distribution of echo amplitude (referred to as tissue texture) applied to digitized images, obtained with commercially available B-mode transcutaneous imaging systems. A total of 47 B-mode images derived from echotomographic studies in 10 patients were digitized off line. For each region of interest, a set of first-order (mean gray level, standard deviation, skewness, kurtosis: mathematical descriptors of the shape of the frequency distribution of gray-level histogram) and of second-order (entropy, angular moment: mathematical descriptors of the spatial distribution of gray levels within the region of interest) textural parameters were evaluated. The visual, concordant reading by two independent, experienced observers assigned the plaques on the basis of qualitatively assessed echodensity to three groups: "soft" (n = 18), "fibrotic" (n = 20), "calcific" (n = 9). Regarding spatial gray-level distribution, 46 plaques would be separated into "homogenous" (n = 17) and "dishomogeneous" (n = 29). On digitized images, the normalized mean gray level was the most effective first-order textural parameter for distinguishing soft (24.2 +/- 12.4 arbitrary units in a zero to 255 scale) from fibrotic (64.5 +/- 16.4) and calcific plaques (125.3 +/- 24.5), P < 0.01 for all intergroup differences. "Homogeneous" plaques were separated from "heterogeneous" ones on the basis of entropy (5 +/- 1 vs 7.9 +/- 9.7; P < 0.01), whereas the values of angular second moment overlapped (1.542E-3 + 1.334E-3 vs 5.181E-4 +/- 2.5615E-4, P = ns). In conclusion, quantitative texture analysis of ultrasonic images derived from transcutaneous, high-resolution, commercially available B-scan systems is feasible in man and provides a quantitative operator-independent assessment of plaque echodensity and homogeneity.

Human cytomegalovirus (HCMV) causes congenital neurological lifelong disabilities. To date, the neuropathogenesis of brain injury related to congenital HCMV (cCMV) infection is poorly understood. This study evaluates the characteristics and pathogenetic mechanisms of encephalic damage in cCMV infection. Ten HCMV-infected human fetuses at 21 weeks of gestation were examined. Specifically, tissues from different brain areas were analyzed by: (i) immunohistochemistry (IHC) to detect HCMV-infected cell distribution, (ii) hematoxylin–eosin staining to evaluate histological damage and (iii) real-time PCR to quantify tissue viral load (HCMV-DNA). The differentiation stage of HCMV-infected neural/neuronal cells was assessed by double IHC to detect simultaneously HCMV-antigens and neural/neuronal markers: nestin (a marker of neural stem/progenitor cells), doublecortin (DCX, marker of cells committed to the neuronal lineage) and neuronal nuclei (NeuN, identifying mature neurons). HCMV-positive cells and viral DNA were found in the brain of 8/10 (80%) fetuses. For these cases, brain damage was classified as mild (n = 4, 50%), moderate (n = 3, 37.5%) and severe (n = 1, 12.5%) based on presence and frequency of pathological findings (necrosis, microglial nodules, microglial activation, astrocytosis, and vascular changes). The highest median HCMV-DNA level was found in the hippocampus (212 copies/5 ng of human DNA [hDNA], range: 10–7,505) as well as the highest mean HCMV-infected cell value (2.9 cells, range: 0–23), followed by that detected in subventricular zone (1.7 cells, range: 0–19). These findings suggested a preferential viral tropism for both neural stem/progenitor cells and neuronal committed cells, residing in these regions, confirmed by the expression of DCX and nestin in 94% and 63.3% of HCMV-positive cells, respectively. NeuN was not found among HCMV-positive cells and was nearly absent in the brain with severe damage, suggesting HCMV does not infect mature neurons and immature neural/neuronal cells do not differentiate into neurons. This could lead to known structural and functional brain defects from cCMV infection. Graphical Abstract

Despite the effectiveness of the currently available antiviral drugs in treating cytomegalovirus (CMV) infection, high rates of adverse effects are associated with their use. Moreover, a problem of increasing importance is the emergence of drug-resistant CMV infection. Here, we describe the first case of off-label use of letermovir (LMV) as preemptive antiviral therapy, in a pediatric allogeneic peripheral blood stem cell transplant recipient with ganciclovir-resistant CMV infection who was intolerant to foscarnet and unable to achieve viral clearance after seven doses of cidofovir. After the administration of LMV, a gradual reduction in viral load was observed and within 6 weeks of LMV treatment, after more than 6 months of positive CMV-DNAemia, the patient cleared the infection. No adverse effects associated with LMV were observed during treatment. In this pediatric study case, the offlabel use of LMV for the treatment of CMV infection has been well tolerated and proved to be effective in leading to the suppression of viral replication.

Introduction:Most infants at risk for cytomegalovirus (CMV)-associated sensorineural hearing loss (SNHL) are unrecognized because of the absence of a universal neonatal CMV screening. The search of CMV-DNA by molecular methods in salivary swabs was demonstrated to be a reliable approach. This study describes the results obtained by carrying out a universal screening for congenital CMV (cCMV) infection including all live-born newborns in three Italian sites, as well as the therapeutic interventions and clinical outcome of the CMV-infected neonates. Moreover, CMV maternal infection's characteristics were evaluated.MethodsTo confirm or exclude cCMV infection, a CMV-DNA-positive result on a first salivary swab was followed by repeated saliva and urine samples collected within 21 days of age. Breast milk samples were also collected. The search of CMV-DNA was performed with a single automated quantitative commercial real-time PCR assay, regardless of the type of samples used.ResultsA total of 3,151 newborns were enrolled; 21 (0.66%) of them were congenitally infected (median saliva viral load at screening, 6.65 [range, 5.03–7.17] log10 IU/ml). Very low/low viral load in screening saliva samples (median value, 1.87 [range, 1.14–2.59] log10 IU/ml) was associated with false-positive results (n = 54; 1.7%). CMV-DNA was detected in almost half of the breast milk samples of mother–infant pairs with a false-positive result, suggesting that contamination from breast milk may not be the only explanation in the study population. cCMV infection confirmation with the search of CMV-DNA in a urine sample proved to be the gold standard strategy, since false-positive results were observed in 4/54 (7.5%) of the repeated saliva samples. Symptomatic cCMV infection was observed in 3/21 (14.3%) infants; notably, one (4.7%) developed moderate unilateral SNHL at 5 months after birth. Finally, two symptomatic cCMV infections were associated with primary maternal infection acquired in the first trimester of gestation; one newborn with severe cCMV symptoms was born to a mother with no CMV checkups in pregnancy.ConclusionWithout universal neonatal CMV screening, some infected infants who develop late neurological sequelae may not be recognized and, consequently, they are not able to benefit early from instrumental and therapeutic interventions to limit and/or treat CMV disease.

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