Human cytomegalovirus (HCMV) leukodnaemia correlates more closely with clinical symptoms than antigenemia and viremia in heart and heart-lung transplant recipients with primary HCMV infection.

BACKGROUND: In the last few years, human cytomegalovirus (HCMV) viremia, pp65 antigenemia, and leuko- and plasma-DNAemia have been developed to quantitate virus in blood of immunocompromised patients with HCMV infection. However, thus far, no conclusive studies have been performed to define the correlation of each of the different assays with clinical symptoms in primary HCMV infections. METHODS: This correlation was investigated in a population of 20 heart and heart-lung transplant recipients with primary HCMV infection using standardized virological methods. RESULTS: Median peak HCMV viremia, antigenemia, and leukoDNAemia levels were 110 (0-2,000) p72-positive fibroblasts, 450 (27-2,000) pp65-positive leukocytes, and >10,000 (1,358-10,000) genome equivalents (GE) in the 14 symptomatic patients and 18 (1-130) p72-positive fibroblasts, 86.5 (5-350) pp65-positive leukocytes, and 248 (10-863) GE in the six asymptomatic patients, respectively. The difference was statistically significant for antigenemia (P=0.009) and leukoDNAemia (P<0.0001). However, on an individual basis, unlike viremia and antigenemia, all DNA peaks of the 6 asymptomatic patients were below the DNA range of the 14 symptomatic patients (<1,000 GE), while all the 14 symptomatic patients had DNA peaks higher than those of asymptomatic patients (>1,000 GE). Follow-up confirmed these results, showing that 1,000-2,000 GE was the threshold zone for emergence of clinical symptoms. Symptoms were never observed in patients with secondary DNA peaks, except for one patient suffering from an HCMV organ localization (HCMV gastritis). CONCLUSIONS: LeukoDNAemia is the viral parameter of choice for monitoring of primary HCMV infections and antiviral treatment in heart and heart-lung transplant recipients. In this patient population, antigenemia-guided preemptive therapy could be replaced by leukoDNAemia-based antiviral therapy.

Circulating cytomegalic endothelial cells are associated with high human cytomegalovirus (HCMV) load in AIDS patients with late-stage disseminated HCMV disease.

The prevalence of circulating cytomegalic endothelial cells, detected currently by the pp65-antigenemia assay and described previously in blood of transplanted and AIDS patients with disseminated human cytomegalovirus (HCMV) infection, was found to be 2.9% in the AIDS population and 6.5% in the fraction of the AIDS population with HCMV in blood. Cytomegalic endothelial cells increased to 39.7% and 48.4%, respectively, in AIDS patients with very high levels of antigenemia and viremia, while an end organ disease reached an incidence of 76.4%. Positive and negative predictive values of cytomegalic endothelial cell detection for diagnosis of HCMV end organ disease were 73.1% and 21.4% with antigenemia levels > 1,000, respectively. On the other hand, in a selected group of 38 cytomegalic endothelial cell-positive AIDS patients with < 50 CD4+ T cells/microliter and late-stage HCMV disease, who were followed-up for variable periods of time, the prevalence of high level antigenemia was 95.3%, that of viremia 86.0% and that of L-DNAemia 92.7%, while the incidence of HCMV end organ disease was 84.2%. In this population, it was shown that cytomegalic endothelial cell presence was associated with lack of (56.0% of episodes) or insufficient (4.0%) anti-HCMV treatment or emergence of HCMV drug-resistant strains (17.3%) or short-term response to antiviral treatment (22.7%); was determined in the same patient by different conditions during follow-up. Longitudinal observations indicated that cytomegalic endothelial cells were detected often in blood at least 3 months later than end organ disease suggesting that the duration of end organ disease was a cofactor associated with the appearance of cytomegalic endothelial cells.

Comparative quantification of human cytomegalovirus DNA in blood of immunocompromised patients by PCR and Murex Hybrid Capture System.

BACKGROUND: Monitoring of human cytomegalovirus (HCMV) load by quantification of antigenemia, viremia and DNAemia is helpful in the management of HCMV infections in immunocompromised patients. In fact, threshold values of these viral parameters are associated with the emergence of clinical symptoms. In addition, the response to antiviral treatment is revealed by a decrease in viral load or virus disappearance from blood. OBJECTIVES: Aim of this study was to compare HCMV DNA quantification in blood of immunocompromised patients by an 'in house' developed quantitative PCR (Q-PCR) assay and the commercially available Murex Hybrid Capture System (HCS). STUDY DESIGN: HCMV DNA was quantified in 95 blood samples from 12 heart and heart-lung transplant recipients and 27 AIDS patients using both techniques. For HCS analysis 3.5 ml whole blood were utilized, whereas Q-PCR was performed using 1 x 10(5) peripheral blood leukocytes (PBL). HCMV DNA levels obtained by HCS and Q-PCR were expressed as number of genome equivalents (GE)/ml whole blood or 1 x 10(5) PBL, respectively. Results from HCS and Q-PCR were compared and submitted to statistical analysis. In addition, HCMV DNA values were compared to levels of antigenemia and viremia. RESULTS AND CONCLUSIONS: Sensitivity of HCS, antigenemia and viremia with respect to Q-PCR were 37.2, 79.5 and 33.3%, respectively. Specificity was 100% for all techniques. On average, samples positive by Q-PCR only, contained low amounts of HCMV DNA. In particular, 45 (91.8%) out of 49 samples negative by HCS and positive by Q-PCR showed < 500 GE/1 x 10(5) PBL. A significant correlation was found between quantitative DNA levels in samples positive by both HCS and Q-PCR (n = 29, R = 0.693, P < 0.01). HCS positivity was associated to significantly higher DNA values as determined by Q-PCR as well as to significantly higher antigenemia and viremia levels. A decrease in DNAemia levels was observed using both HCS and Q-PCR after antiviral treatment. Given that the great majority of blood samples missed by HCS contain low levels of HCMV DNA which are not clinically significant, HCS seems very promising as an alternative to HCMV DNA quantification by PCR in solid organ transplant recipients and AIDS patients.

Fibrinolytic parameters in patients undergoing total hip replacement: relationship with the development of asymptomatic deep vein thrombosis and diagnostic usefulness of venous occlusion.

BACKGROUND AND OBJECTIVE: It has been suggested that impaired fibrinolytic activity contributes to deep vein thrombosis in orthopedic surgery. Studying the fibrinolytic system following venous occlusion has been proposed as a good method of detecting the risk of this postoperative complication. The objective of this work was to verify whether venous occlusion represents a reliable method of detecting an impaired fibrinolytic response after total hip replacement. METHODS: Thirty-two consecutive patients undergoing total hip replacement were studied. Citrated blood samples were taken from each patient the day before surgery and on postoperative days 1, 3, and 7, before and after venous occlusion, in order to evaluate plasma levels of tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI-1). All patients underwent bilateral phlebography 10 days after surgery. RESULTS: Seven out of 32 patients (21.9%) developed deep venous thrombosis (DVT) according to the venographic test. After surgery, an increase in t-PA antigen levels was detected both in patients who developed DVT and in those who did not, with a significant increase on the first and seventh days after surgery only in the non-DVT group. After 10-min venous occlusion, t-PA antigen levels increased at all postoperative recordings in both groups of patients, but most significantly on days 1 and 7 after surgery. PAI-1 antigen plasma levels, when measured before venous occlusion, increased only in non-DVT patients on the seventh postoperative day. After venous occlusion, a difference was found between the two groups only postoperatively on day 7 with regard to PAI-1 levels. INTERPRETATION AND CONCLUSIONS: According to our results, no impaired fibrinolytic response was found in DVT patients. In addition, venous stasis seems to give no further information with respect to basal values in the early detection of postoperative thromboembolic complications in orthopedic patients.

[Histoenzymologic data on the epithelial cells of the gastric mucosa of marmosets (Callithrix jacchus & Callithrix penicillata)]. / Dados histoenzimoógicos sobre as células epiteliais da mucosa gástrica de saguis (Callithrix jacchus & Callithrix penicillata).

The gastric mucosa of marmosets is devoid of UDPG-GT; phosphorylases; G-6-PA; F-1,6-PA; alanyl aminopeptidase and leucine aminopeptidase. Only the acid phosphatase was seen with a stronger reactivity in the chief cells. The other enzymes (LDH; G-6-PDH; 6-PGDH; NADPH2-TR; cis-aconitase; ICDH; SDH; MDH; cytochrome oxidase; NADH2-TR; a-GPDH; b-OHBDH and nonspecific esterase) showed a stronger reactivity in the parietal cells.