Daniela Riggio

Typing of human papillomavirus in women with cervical lesions: Prevalence and distribution of different genotypes

Human papillomavirus (HPV) are distributed widely and persistent infection with high‐risk (HR) HPV is recognized as a necessary cause of cervical cancer. The aim of this study was to evaluate the distribution of different HR‐HPV genotypes in 199 women with cervical pre‐invasive lesions undergoing conservative treatment. A Linear Array HPV Genotyping Test was used to identify individual HPV genotypes in cervical samples. It was observed that the most prevalent HPV genotypes were HPV 16 (52.6%), HPV 51 (13.5%), and HPV 31 (10.9%); HPV 18 was found in 7.3% of the patients. Stratifying the different HPV genotypes according to the severity of the cervical lesion, a strong association between the increasing severity of the histological diagnosis and the detection of more carcinogenic HR‐HPV type was found, and in all but one cervical intraepithelial neoplasia of grade 3 the presence of at least one HR‐HPV could be detected, with more than 70% of cervical intraepithelial neoplasia of grade 3 patients bearing HPV 16. Multiple infections, comprising between 2 and 6 HPV types, were found in 43% of patients; however, the presence of more than 1 HR‐HPV type was not associated with an increased risk of high grade lesions. In conclusion, this data show that HPV 16, 51, 31, 52, and 18 were the prevalent types found in patients with cervical lesion undergoing conservative treatment, with a high prevalence of HPV 16 in cervical intraepithelial neoplasia of grade 3 patients. No association between multiple infection and severity of the lesion could be found. J. Med. Virol. 81:271–277, 2009.

Comparison of the clinical performance of carcinogenic HPV typing of the Linear Array and Papillocheck® HPV-screening assay.

BACKGROUND HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 66 are considered carcinogenic for human beings. DNA-chip technology, Papillocheck HPV-screening (Greiner) and reverse dot blot, Linear Array (LA) (Roche) are tools to assess the distribution of HPV genotypes. OBJECTIVES The aim of the study was to compare the clinical performance of Papillocheck and LA assays using a clinical cut-off of CIN2+. The secondary aim was to comparatively assess the distribution of HPV types using these two assays. STUDY DESIGN The study population comprised 239 women referred for colposcopy and histology. Papillocheck, LA, and Hybrid Capture II (HCII) tests were done on all samples. RESULTS All tests showed good sensitivity and NPV (greater than 90%). None of the comparisons of sensitivities, specificities, PPVs, and NPVs showed statistically relevant differences between tests. High-risk HPV positivity rate was similar for all tests (Papillocheck 75%, LA 77%, and HCII 73%). Agreement between tests was good. The concordance levels between HCII and Papillocheck and between HCII and LA were 93% (k=0.82) and 92% (k=0.80), respectively. Papillocheck and LA tests showed a high overall concordance rate of 96% (k=0.90). HPV16 was the most detected type (45% with Papillocheck, and 47% with LA), and HPV31 was the second most detected type (13% with Papillocheck, and 14% with LA). CONCLUSIONS The Papillocheck HPV-screening test and LA test have a good clinical sensitivity to detect HPV types in CIN2+ patients. These assays allow, in the same experiment, to detect and determine the virus type. Our study showed that HPV types 16 and 31/33 are the most prevalent.

Interchangeability of measurements of CA 19-9 in serum with four frequently used assays : an update

Abstract Background: CA 19-9 is a marker principally related to pancreatic and gall bladder cancer. Although its determination has no value in screening for these malignancies, it is used in post-operative monitoring and during chemotherapeutic treatment of confirmed disease. Measurements during follow-up must be comparable and must be performed with standard, validated methods. Methods: We compared four routinely used analytical systems for CA 19-9 determination: the Architect i2000 and AxSYM systems from Abbott Laboratories, the Elecsys 1010 from Roche Diagnostics, and the KRYPTOR system from Brahms Diagnostics. We evaluated the analytical performance of the four systems and compared measurements of CA 19-9 values, which covered the whole analytical range. Results: The analytical performance and accuracy of the four systems were fairly good, but Passing-Bablok regression and mountain plots showed significant differences in CA 19-9 values measured with the four platforms. Conclusions: Our data indicate that during tumor follow-up, the use of the same system is appropriate to avoid the risk of a variation due to the method rather than the disease. Moreover, whenever a change in analytical equipment is required, careful analysis of CA 19-9 results must be undertaken. Clin Chem Lab Med 2007;45:100–4.

Interference of antibiotic therapy on blood cultures time-to-positivity: analysis of a 5-year experience in an oncological hospital.

This study performed a retrospective analysis on the relationship between blood culture time-to-positivity (TP) and type of isolated microorganism, antibiotic administration, and immunological status of the patients. We analyzed the data related to 1,218 positive blood cultures. When compared to Gram positive bacteraemia, the percentage of Gram negative growth was higher and the mean TP significantly shorter (p < 0.0001). In patients receiving antibiotics, median and mean TPs of blood culture were different for Gram positive bacteraemia (log-rank p = 0.0022, Wilcoxon p < 0.0001) but not for Gram negative (log-rank p = 0.4011, Wilcoxon p = 0.1585). No statistically significant effect on TP was found for sampling site, interaction between sampling site and antibiotic administration, and immunological status of the patient. In conclusion, TP is independent of antibiotic therapy in cases of Gram negative bacteraemia, while for Gram positive bacteraemia a prolongation of TP occurs.

Atrial fibrillation after thoracic surgery for lung cancer: use of a single cut-off value of N-terminal pro-B type natriuretic peptide to identify patients at risk

Postoperative atrial fibrillation (AF) is a well-known complication occurring after thoracic surgery. B-type natriuretic peptide has recently been investigated as a predictive marker of postoperative AF after cardiac surgery. The aim of this study was to evaluate a definite cut-off for N-terminal pro-B type natriuretic peptide (NT-proBNP) in predicting postoperative AF in lung cancer patients. NT-proBNP was determined before and after surgery in 400 patients. Cardiac function was monitored by continuous postoperative ECG and clinical cardiological evaluation. AF occurred in 18% of the patients. Receiver operating characteristic curve analyses identified a cut-off of 182.3 ng l−1 as the one with the highest sensitivity and specificity. Perioperative increased levels of NT-proBNP seem to predict postoperative AF in patients undergoing thoracic surgery, and a single cut-off of 182.3 ng l−1 can be used to select high-risk patients who could receive preventive therapy, leading to a considerable decrease in the total costs associated with the management of this complication.

MiR-320a as a Potential Novel Circulating Biomarker of Arrhythmogenic CardioMyopathy

Diagnosis of Arrhythmogenic CardioMyopathy (ACM) is challenging and often late after disease onset. No circulating biomarkers are available to date. Given their involvement in several cardiovascular diseases, plasma microRNAs warranted investigation as potential non-invasive diagnostic tools in ACM. We sought to identify circulating microRNAs differentially expressed in ACM with respect to Healthy Controls (HC) and Idiopathic Ventricular Tachycardia patients (IVT), often in differential diagnosis. ACM and HC subjects were screened for plasmatic expression of 377 microRNAs and validation was performed in 36 ACM, 53 HC, 21 IVT. Variable importance in data partition was estimated through Random Forest analysis and accuracy by Receiver Operating Curves. Plasmatic miR-320a showed 0.53 ± 0.04 fold expression difference in ACM vs. HC (p < 0.01). A similar trend was observed when comparing ACM (n = 13) and HC (n = 17) with athletic lifestyle, a ACM precipitating factor. Importantly, ACM patients miR-320a showed 0.78 ± 0.05 fold expression change vs. IVT (p = 0.03). When compared to non-invasive ACM diagnostic parameters, miR-320a ranked highly in discriminating ACM vs. IVT and it increased their accuracy. Finally, miR-320a expression did not correlate with ACM severity. Our data suggest that miR-320a may be considered a novel potential biomarker of ACM, specifically useful in ACM vs. IVT differentiation.

B-type natriuretic peptide levels in patients with pericardial effusion undergoing pericardiocentesis

OBJECTIVES Pericardial effusion is characterized by progressive accumulation of fluid within the pericardial space, resulting in increased intra-pericardial pressure and compression of the heart. As B-type natriuretic peptide (BNP) is secreted by the ventricles in response to increased myocardial stretch, we hypothesized that pericardial effusion, as well as its resolution, might influence BNP plasma levels. METHODS We prospectively measured, in 146 consecutive patients with pericardial effusion, BNP plasma levels at baseline, soon after, and 24h after pericardiocentesis. A scoring system based on 7 clinical and echocardiographic parameters was developed, and patients were classified according to the number of variables as having low (0-2), intermediate (3-4), or high (5-7) severity score. RESULTS Out of the 146 patients, 42 (29%) had normal values (<100pg/ml), whereas 104 (71%) had high BNP values at baseline. In the whole population, baseline BNP levels significantly decreased as the severity score increased (r=-0.21; P=0.01). 24h after pericardiocentesis, a significant increase in BNP was observed in patients with intermediate (P=0.004) score and with high (P<0.001) severity score; no increase occurred in low score patients (P=0.56). The higher was the severity score, the steeper was the increase in BNP through the three time-points considered (P=0.04). CONCLUSIONS The results of the present study show that BNP plasma levels are suppressed in the presence of severe pericardial effusion, and that they rise after pericardiocentesis. Future studies should investigate the role of BNP in assisting clinicians in the decision-making process of pericardial fluid drainage.

Laboratory-based management of microbiological alerts: effects of an automated system on the surveillance and treatment of nosocomial infections in an oncology hospital.

Background: Prevention and surveillance programs are key to contain Nosocomial Infections (Nis). At the European Institute of Oncology, surveillance based on ex-post data collection has been done since the inception of hospital activity; laboratory-based surveillance of microbiological alert was not standardized. This study describes the issues related to the recent introduction into the hospital routine of a laboratory-based automated surveillance system and its clinical impact on monitoring and treatment of Nis. Methods: An interdisciplinary team defined the alerts and the actions to be taken in response; recipients of the alert messages were identified and software was programmed. Program features were created so their employment would generate a prompt notification of clinically critical results. After a training period, the program was introduced in the hospital routine. Results: There were a total of 150 generated alerts; the main alert related to microorganisms requiring prompt patient isolation and/or public notification. Clinical use of the program was relevant in detection and immediate notification of Cytomegalovirus active infection in stem cell recipients and central venous catheter related candidemia: the prompt administration of adequate treatment was possible hours earlier compared to the previous approach. Conclusions: A laboratory-based automated surveillance system is effective in facilitating the management of Nis; its clinical employment also leads to important clinical advantages in patient care.

ST2 and B-type natriuretic peptide kinetics during exercise in severe heart failure: ST2 and B-type natriuretic peptide kinetics during exercise in severe heart failure

In chronic heart failure (HF), risk stratification is one of the most challenging issues, with a plethora of biomarkers available for this purpose. The key role is played by B-type natriuretic peptide (BNP) and/or its amino-terminal fragment NT-proBNP.1 Accordingly, all new biomarkers should be compared with BNP and/or should be characterized by prognostic capability in selected HF populations. Soluble ST2 has recently been introduced among the new biomarkers. The presence of high levels of ST2 is related to the severity of HF and to an increased risk of complications, such as arrhythmias, acute decompensation, and death, independently of natriuretic peptides.2 Indeed, a prognostic role of ST2 has been suggested in low-risk populations,3 in patients with chronic,4 advanced, and acute2 HF. Regardless, at present, the most defined role of ST2 is to provide a prognostic value additive to that of NT-proBNP within a multiparametric prognostic approach.5 The changes in ST2 in response to an acute haemodynamic event are presently unknown, and specifically it is unknown whether ST2 response to an acute haemodynamic event has the same magnitude and time frame of that reported for BNP. Indeed, in patients with severe HF, BNP increases even during a 10 min maximal exercise, probably mirroring acute pulmonary haemodynamic worsening.6 We studied 30 (67±10 years, 28 male) consecutive patients with chronic severe HF (left ventricular ejection fraction 29± 9%) who belonged to an HF population regularly followed at our HF unit. Inclusion criteria were peak oxygen uptake (VO2) <12 mL/kg/min, optimized HF therapy, stable clinical conditions, left ventricular ejection fraction (echocardiography) <45%, and capability to perform spirometry and lung diffusion test. The locally appointed ethics committee approved the research protocol (R115/14-CCM), and informed consent was obtained from all patients. Patients underwent a 10 min cycleergometer ramp cardiopulmonary exercise test protocol. Immediately before exercise and at peak exercise, venous blood was sampled for BNP and ST2 determination. At the same time, lung diffusion (DLCO) and its two components membrane diffusion (DM) and capillary volume (Vcap) were measured. DLCO was measured by the single-breath constant expiratory flow technique (Sensor Medics 2200, Yorba Linda, CA, USA). DLCO subcomponents, Vcap and DM, were calculated applying the Roughton and Forster method. Soluble ST2 levels were assessed using a highly sensitive sandwich monoclonal immunoassay (Presage® ST2 Assay, Critical Diagnostics, San Diego, CA, USA). BNP was measured using the chemiluminescent microparticle immunoassay on the Architect i2000 analyser (Abbott Diagnostics, Abbott Park, IL, USA). Data confirmed severe exercise limitation, since peak VO2 was 855± 224 mL/min, equivalent to 10.8±1.6 mL/kg/min and to 45± 8% of the predicted value, VO2 at anaerobic threshold was 8.0±1.4 mL/kg/min, and ventilation/carbon dioxide production relationship slope was 38±10. DLCO, BNP and ST2 at rest and peak exercise are reported in Table 1. BNP (rest–peak +15±16%, P< 0.05), but not ST2 (rest–peak 1±12%), significantly increased at peak exercise, showing that a transitory haemodynamic impairment