Clinical response to biologicals for severe asthma: any relevance for sex in different age ranges?

Background: Whether sex can influence the clinical response to biological treatment in patients with severe asthma has not been fully addressed. Aims and methods: The aim of this study was to investigate in patients with severe asthma undergoing biological treatment the individual evolution of lung function measurements and patient-reported asthma control scores over a 12-month follow-up period, in relation to patients' sex, in different age ranges. Second, the change in the administered dose of oral corticosteroids (OCS) before and after 12 months of treatment was investigated. Results: 64 patients (58% female and 42% male) with a median age of 52 years were enrolled in the study. There were no relevant differences between sexes in terms of lung function, patient-reported asthma control, exacerbation rate and daily OCS dose within the study timeframe. A separate sub-analysis by biological treatment confirmed the same finding. Stratifying individuals by age, we showed that older men had lower lung function parameter values (forced expiratory volume in 1 s (FEV1) % predicted and FEV1/forced vital capacity index) than older women, whereas an opposite trend was observed in terms of Asthma Control Test score. No other relevant differences were detected after age stratification. Conclusion: According to our findings, sex does not act as a determinant of treatment response to biologicals in people with severe asthma. Although to be confirmed in larger studies, our data suggest that neither sex nor age should limit biological treatment prescription, once the eligibility criteria for that therapy are satisfied.

Patient Safety in the Eyes of Aspiring Healthcare Professionals: A Systematic Review of Their Attitudes.

A culture of safety is important for the delivery of safe, high-quality care, as well as for healthcare providers' wellbeing. This systematic review aimed to describe and synthesize the literature on patient safety attitudes of the next generation of healthcare workers (health professional students, new graduates, newly registered health professionals, resident trainees) and assess potential differences in this population related to years of study, specialties, and gender. We screened four electronic databases up to 20 February 2020 and additional sources, including weekly e-mailed search alerts up to 18 October 2020. Two independent reviewers conducted the search, study selection, quality rating, data extraction, and formal narrative synthesis, involving a third reviewer in case of dissent. We retrieved 6606 records, assessed 188 full-texts, and included 31 studies. Across articles, healthcare students and young professionals showed overwhelmingly positive patient safety attitudes in some areas (e.g., teamwork climate, error inevitability) but more negative perceptions in other domains (e.g., safety climate, disclosure responsibility). Women tend to report more positive attitudes. To improve safety culture in medical settings, health professions educators and institutions should ensure education and training on patient safety.

Prevalence of SARS-CoV-2, Verona, Italy, April-May 2020.

We used random sampling to estimate the prevalence of severe acute respiratory syndrome coronavirus 2 infection in Verona, Italy. Of 1,515 participants, 2.6% tested positive by serologic assay and 0.7% by reverse transcription PCR. We used latent class analysis to estimate a 3.0% probability of infection and 2.0% death rate.

Prevalence of asymptomatic SARS-CoV-2-positive individuals in the general population of northern Italy and evaluation of a diagnostic serological ELISA test: a cross-sectional study protocol.

INTRODUCTION: As of 30 April 2020, the novel betacoronavirus SARS-CoV-2 had infected more than 3 172 000 individuals, killing over 224 000 people and spreading to more than 200 countries. Italy was the most affected country in Europe and the third most affected in the world in terms of the number of cases. Therefore, the aims of this study are: (1) to estimate the prevalence of asymptomatic SARS-CoV-2-positive individuals among the general population of Verona; (2) to assess the accuracy (sensitivity, specificity and predictive values) of an ELISA serological test for the screening of SARS-CoV-2. METHODS AND ANALYSIS: The study will be carried out on a random sample of subjects aged at least 10 years from the general population of Verona. Participants will undergo the measurement of vital parameters (oxygen saturation measured by oximeter, respiratory rate and body temperature detected by laser thermometer), the administration of a COVID-19-related symptoms questionnaire, the collection of a blood sample and a nasopharyngeal swab. Our evaluation will include the statistical technique of Latent Class Analysis, which will be the basis for the estimation of prevalence. ETHICS AND DISSEMINATION: The study protocol has been approved by the Ethics Committee of Verona and Rovigo provinces on 15 April 2020 (internal protocol number 2641CESC). The study results will be submitted for publication in international, peer-reviewed journals and the complete dataset will be deposited in a public repository. Most relevant data will be made available to policy-makers as well as disseminated to stakeholders and to the community.

Enhancer of zeste 2 polycomb repressive complex 2 subunit polymorphisms in melanoma skin cancer risk.

Melanoma is the most deadly skin cancer, and its incidence is growing. EZH2, a member of the Polycomb Group (PcGs) proteins family, plays an important biological role in the occurrence and development of melanoma. EZH2 germline genetic polymorphisms have not been yet evaluated in melanoma predisposition. Three hundred thirty sporadic Italian melanoma patients and 333 healthy volunteers were genotyped to analyse the association between EZH2 variants rs6950683, rs2302427, rs3757441, rs2072408 and melanoma risk. The functionality of rs6950683 alleles was investigated in keratinocytes (HaCat), melanoma cells (A375) and human embryonic kidney cells (HEK293), using promoter-reporter assays. Genotype distribution of SNPs showed that rs6950683T and rs3757441C alleles were positively associated with melanoma risk (P = .003 and .004, respectively). Haplotype analysis revealed that TCCA and CCCG haplotypes were associated with a higher risk of melanoma (P = .02 and .04, respectively). Functional assays demonstrated that allele rs6950683T reduce promoter activity in the three cell lines analysed compared to C allele. rs6950683T and rs3757441C alleles in the EZH2 gene appear positively associated with melanoma risk in the analysed population. In addition, we demonstrated for the first time the functional role of rs6950683 upstream polymorphism on EZH2 gene expression regulation.

Attitudes of Nursing Home Staff Towards Influenza Vaccination: Opinions and Factors Influencing Hesitancy.

Seasonal influenza is recognized to be a significant public health problem and a cause of death, especially in fragile persons. In nursing homes (NHs), vaccination for both residents and staff is the best preventive strategy. However, professionals' immunization rates are far from reaching the international recommended values. This study aims to describe the adherence and attitudes of NH staff towards flu vaccination and to explore staff hesitancy. A questionnaire was developed based on a literature review and on the 3Cs (confidence, complacency, convenience) of the WHO framework and administered among the staff of four NHs of a province in the northeast of Italy. Results demonstrated a low adherence towards annual vaccination (i.e., only 3% declared getting the flu vaccination each year). Complacency, confidence and convenience all showed a significant impact on the attitude towards vaccination both in univariate and multivariable analysis, with complacency being the most strongly associated area. The area of confidence resulted in strongly challenging factors. Only 24.8% of interviewees appeared trustful towards the efficacy of receiving immunization and 34% declared safety issues. Insights from the study can support the implementation of effective interventions to improve vaccination adherence in NHs. Specifically, increasing complacency by raising awareness related to the risks of influenza appears to be an essential strategy to effectively promote vaccination uptake.

Screening for hemodialysis graft stenosis and short-term thrombosis risk: A comparison of the available tools.

INTRODUCTION: Guidelines recommend hemodialysis graft screening to identify and repair significant (>50%) stenosis at high risk of thrombosis, but there is insufficient evidence to prefer one or other screening tool due to the lack of studies comparing all available options. METHODS: Seeking an optimal screening approach, we compared the performance of all currently used tools (duplex ultrasound to detect significant stenosis (StD) and measure access blood flow (QaD), ultrasound dilution access blood flow (QaU), static venous intra-access pressure ratio (VAPR), dynamic arterial and venous pressures measurement, and monitoring) for diagnosing significant angiography-proven stenosis (StA) and predicting incipient thrombosis (occurring within 4 months) in 62 grafts. All thrombotic and symptomatic acute hypotension episodes were recorded during follow-up. RESULTS: VAPR > 0.70 and QaU < 1600 mL/min were the best indicators to angiography for those aiming to identify the majority of StA (91% sensitivity) and QaU < 1000 mL/min or StD for those aiming to avoid unnecessary angiograms (95%-93% positive predictive value). At Cox's analysis, the only significant thrombosis predictors were acute hypotension episodes (relative risk = 4.4 (95% confidence interval = 2.2-8.8), p < 0.0001) and QaU or QaD (14% (95% confidence interval = 8-21) or 16% (95% confidence interval = 6-25) increased risk per 100 mL/min drop in Qa, p < 0.003). Thrombosis risk (adjusted for acute hypotension) became significantly higher at QaU = 1000-700 mL/min (relative risk = 3.6 (95% confidence interval = 1.6-8.2), p < 0.001) and QaD = 1300-1000 mL/min (relative risk = 3.1 (95% confidence interval = 1.1-12.8), p = 0.031). The proportion of thromboses attributable to acute hypotension was 40% (95% confidence interval = 24-57). CONCLUSIONS: Our comparative study showed that an effective screening for graft stenosis and short-term thrombosis risk can rely on Qa surveillance alone, and suggested that avoiding acute hypotension and correcting stenosis at QaU < 1000 mL/min or QaD < 1300 mL/min can contain thrombosis risk.

Serum miR-223: A Validated Biomarker for Detection of Early-Stage Non-Small Cell Lung Cancer.

BACKGROUND: The published circulating miRNA signatures proposed for early-stage non-small cell lung cancer (NSCLC) detection are inconsistent and difficult to replicate. Reproducibility and validation of an miRNA simple signature of NSCLC are prerequisites for translation to clinical application. METHODS: The serum level of miR-223 and miR-29c, emerging from published studies, respectively, as a highly sensitive and a highly specific biomarker of early-stage NSCLC, was measured with droplet digital PCR (ddPCR) technique in an Italian cohort of 75 patients with stage I-II NSCLC and 111 tumor-free controls. By ROC curve analysis we evaluated the miR-223 and miR-29c performance in discerning NSCLC cases from healthy controls. RESULTS: Reproducibility and robust measurability of the two miRNAs using ddPCR were documented. In a training set (40 stage I-II NSCLCs and 56 controls), miR-223 and miR-29c, respectively, showed an AUC of 0.753 [95% confidence interval (CI), 0.655-0.836] and 0.632 (95% CI, 0.527-0.729) in identifying NSCLC. Combination of miR-223 with miR-29c yielded an AUC of 0.750, not improved over that of miR-223 alone. Furthermore, in an independent blind set (35 stage I-II NSCLCs and 55 controls), we validated serum miR-223 as an effective biomarker of stage I-II NSCLC (AUC = 0.808; 95% CI, 0.712-0.884), confirming the miR-223 diagnostic performance reported by others in Chinese cohorts. CONCLUSIONS: Using ddPCR technology, miR-223 was externally validated as a reproducible, effective serum biomarker of early-stage NSCLC in ethnically different subjects. Combination with miR-29c did not improve the miR-223 diagnostic performance. IMPACT: Serum miR-223 determination may be proposed as a tool for refining NSCLC risk stratification, independent of smoking habit and age.

Knowledge and use of e-cigarettes among nursing students: results from a cross-sectional survey in north-eastern Italy.

BACKGROUND: Data on electronic cigarette (e-cigarette) use among health professional students, who can play a central role in promoting healthy habits and smoking cessation, are sparse. Moreover, the association between e-cigarettes and smoking habits is still debated. The present study aimed to investigate the diffusion of e-cigarette use among nursing students in north-eastern Italy and explore its association with tobacco smoking. METHODS: In 2015, a questionnaire focused on e-cigarette use and tobacco smoking habits was anonymously administered to 2020 students attending nursing courses held by Verona University in 5 different centres. Of these students, 1463 (72.4%) answered the questionnaire. The influence of e-cigarette ever use on both tobacco smoking initiation in all subjects and smoking cessation among ever smokers was investigated by multivariable logistic models. RESULTS: Most responders were female (77.1%), and the mean (SD) age was 23.2 (4.2) years. Nearly all students (94.7%) had heard about e-cigarettes. Approximately one-third (30.3, 95% CI 27.9-32.7%) had ever used e-cigarettes, but only 2.1% (1.5-3.0%) had used e-cigarettes in the last month. Very few (2.1%) of those responders who had never used e-cigarettes were willing to try them. Prevalence values were much higher for tobacco smoking: 40.9% of responders reported being current tobacco smokers, and 10.1% reported being past smokers. Ever use and current use of e-cigarettes were reported by 57.2 and 4.4% of current tobacco smokers and by 12.0 and 0.6% of never or past smokers, respectively (p < 0.001). In multivariable analysis, students who ever used e-cigarettes had 13 times greater odds of being an ever tobacco smoker than never users, whereas they had three times lower odds of being a former smoker. Only 26 students were currently using both electronic and tobacco cigarettes, and most declared that they used e-cigarettes to stop or reduce tobacco smoking. Of note, only three students reported that they had completely stopped smoking thanks to e-cigarette use. CONCLUSION: Use of e-cigarettes seemed to be rather rare among Italian nursing students and was mainly restricted to current smokers. E-cigarette use was not associated with smoking cessation in nursing students.

Reducing interruptions during medication preparation and administration.

PURPOSE: According to literature, interruptions during drug administration lead to a significant proportion of medication errors. Evidence on the effectiveness of interventions to reduce interruption is still limited. The purpose of this paper is to explore main reasons for interruptions during drug administration rounds in a geriatric ward of an Italian secondary hospital and test the effectiveness of a combined intervention. DESIGN/METHODOLOGY/APPROACH: This is a pre and post-intervention observational study based on direct observation. All nurse staff (24) participated to the study that lead to observe a total of 44 drug dispensing rounds with 945 drugs administered to 491 patients in T0 and 994 drugs to 506 patients in T1. FINDINGS: A significant reduction of raw number of interruptions (mean per round from 17.31 in T0 to 9.09 in T1, p<0.01), interruptions/patient rate (from 0.78 in T0 to 0.40 in T1, p<0.01) and interruptions/drugs rate (from 0.44 in T0 to 0.22 in T1, p<0.01) were observed. Needs for further improvements were elicited (e.g. a greater involvement of support staff). PRACTICAL IMPLICATIONS: Nurse staff should be adequately trained on the risks related to interruptions during drug administration since routine activity is at high risk of distractions due to its repetitive and skill-based nature. ORIGINALITY/VALUE: A strong involvement of both MB and leadership, together with the frontline staff, helped to raise staff motivation and guide a bottom-up approach, able to identify tailored interventions and serve concurrently as training instrument tool.

Lack of association of metastasis-associated lung adenocarcinoma transcript 1 variants with melanoma skin cancer risk.

The long noncoding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) has been implicated in melanoma. Polymorphisms in MALAT1 may play a vital role in the progress of melanoma by its regulative function. However, potential genetic variants in MALAT1 affecting the risk of melanoma onset have not been explored. In this study, two single nucleotide polymorphisms (rs3200401 and rs619586) in MALAT1 were selected for genotyping of 334 melanoma patients and 291 cancer-free controls in an Italian population. The results showed that MALAT1 rs3200401 and rs619586 were not associated with melanoma risk. A further breakdown analysis by sex stratification also indicated a lack of association between these polymorphisms and melanoma. In addition, we tested 450 bp of the proximal 5´ flanking region of the gene for the presence of polymorphisms that could be associated with melanoma risk and found no variants in 96 melanoma patients. In conclusion, our results suggest that there is no contribution of MALAT1 rs3200401 and rs619586 polymorphisms or polymorphisms in the core promoter that could be associated with the risk of melanoma skin cancer in this specific study setting. Further validation will be required in larger studies involving different settings/larger populations in order to reach conclusive results.

Pro: Vascular access surveillance in mature fistulas: is it worthwhile?

Guidelines recommend regular screening of mature arteriovenous fistulas (AVFs) for preemptive repair of significant stenosis (≥50% lumen reduction) at high risk of thrombosis, identifiable from clinical signs of access dysfunction (monitoring) or by measuring access blood flow (Qa surveillance), which also enables stenosis detection in functional accesses. To compare the value of Qa surveillance versus monitoring, a meta-analysis was performed on the randomized controlled trials (RCTs) comparing the two screening strategies. It emerged that correcting stenosis identified by Qa surveillance significantly halved the risk of thrombosis [relative risk (RR) = 0.51, 95% confidence interval (CI) 0.35-0.73] and access loss (RR = 0.47, 95% CI 0.28-0.80) in comparison with intervention prompted by clinical signs of access dysfunction. One small RCT aiming to identify an optimal Qa threshold showed that stenosis repair at Qa >500 mL/min produced a significant 3-fold reduction in the risk of thrombosis (RR = 0.37, 95% CI 0.12-0.97) and access loss (RR = 0.36, 95% CI 0.09-0.99) in comparison with intervening when Qa dropped to <400 mL/min as per guidelines. To test the real-world benefits of Qa surveillance, the expected RCT-based thrombosis and access loss rates with Qa surveillance were compared with the rates with monitoring reported in observational studies: the expected thrombosis and access loss rates with surveillance were only lower than with monitoring when a Qa >500 mL/min was considered (2.4, 95% CI 1.0-4.6 and 2.2, 95% CI 0.7-5.0 versus 9.4, 95% CI 7.4-11.3 and 10.3, 95% CI 7.7-13.4 events per 100 AVFs-year, P ≤ 0.024), suggesting that in clinical practice adopting Qa surveillance may only be worthwhile at centres with high thrombosis and access loss rates associated with monitoring, and adopting Qa thresholds >500 mL/min for elective stenosis repair.

A single dialysis session of hemodiafiltration with sorbent-regenerated endogenous ultrafiltrate reinfusion (HFR) removes hepcidin more efficiently than bicarbonate hemodialysis: a new approach to containing hepcidin burden in dialysis patients?

BACKGROUND: Most hemodialysis patients have high Hepcidin-25 levels, which may be involved in the pathogenesis of several uremic complications related to an altered iron biology. The hemodialysis procedure itself can influence Hepcidin-25 levels by removing Hepcidin-25 and maybe stimulating its production due to a pro-inflammatory effect. METHODS: To assess the relationship between dialysis-related inflammation and intradialysis changes in Hepcidin-25, we performed a crossover trial in 28 hemodialysis patients to compare the effects on serum levels of Hepcidin-25 and inflammatory markers activated during dialysis [Tumor Necrosis Factor-α (TNF-α), Interleukin-6, C-reactive protein (CRP), Pentraxin-3] of a single dialysis session using a technique capable of reducing inflammation, HFR (Hemo Filtrate Reinfusion: a hemodiafiltration system combining convection, diffusion and adsorption) or bicarbonate-dialysis using either the same low-flux membrane as in the diffusion stage of HFR (LFBD) or a high-flux membrane (HFBD). RESULTS: HFR achieved a greater reduction in Hepcidin-25 levels than both LFBD [-72% (95% CI: -11 to -133), p = 0.022] and HFBD [-137% (95% CI: -2 to -272), p = 0.047], conceivably due to both a greater removal (because of its convective/adsorptive component) and a lower inflammation-related Hepcidin-25 production. HFR also led to a greater decrease in TNF-α than LFBD [-277% (95% CI: -59 to -494), p = 0.014], while the two methods induced similar changes in Interleukin-6, CRP and Pentraxin-3 levels. CONCLUSIONS: Our findings suggest that a single bicarbonate-dialysis session can upregulate Hepcidin-25 synthesis and that HFR can fully overcome this effect, enabling a greater Hepcidin-25 removal during dialysis. Adequately-designed studies are needed, however, to establish whether the beneficial effect of HFR emerging from our study could reduce Hepcidin-25 (and TNF-α) burden and improve clinically-relevant outcomes. TRIAL REGISTRATION: ISRCTN15957905.

Systematic review and critique of circulating miRNAs as biomarkers of stage I-II non-small cell lung cancer.

Selected circulating microRNAs (miRNAs) have been suggested for non-invasive screening of non-small cell lung cancer (NSCLC), however the numerous proposed miRNA signatures are inconsistent. Aiming to identify miRNAs suitable specifically for stage I-II NSCLC screening in serum/plasma samples, we searched the databases "Pubmed", "Medline", "Scopus", "Embase" and "WOS" and systematically reviewed the publications reporting quantitative data on the efficacy [sensitivity, specificity and/or area under the curve (AUC)] of circulating miRNAs as biomarkers of NSCLC stage I and/or II. The 20 studies fulfilling the search criteria included 1110 NSCLC patients and 1009 controls, and were of medium quality according to Quality Assessment of Diagnostic Accuracy Studies checklist. In these studies, the patient cohorts as well as the control groups were heterogeneous for demographics and clinicopathological characteristics; moreover, numerous pre-analytical and analytical variables likely influenced miRNA determinations, and potential bias of hemolysis was often underestimated. We identified four circulating miRNAs scarcely influenced by hemolysis, each featuring high sensitivity (> 80%) and AUC (> 0.80) as biomarkers of stage I-II NSCLC: miR-223, miR-20a, miR-448 and miR-145; four other miRNAs showed high specificity (> 90%): miR-628-3p, miR-29c, miR-210 and miR-1244. In a model of two-step screening for stage I-II NSCLC using first the above panel of serum miRNAs with high sensitivity and high AUC, and subsequently the panel with high specificity, the estimated overall sensitivity is 91.6% and overall specificity is 93.4%. These and other circulating miRNAs suggested for stage I-II NSCLC screening require validation in multiple independent studies before they can be proposed for clinical application.

Sex-specific effect of RNASEL rs486907 and miR-146a rs2910164 polymorphisms' interaction as a susceptibility factor for melanoma skin cancer.

The genetics of melanoma is complex and, in addition to environmental influences, numerous genes are involved or contribute toward melanoma predisposition. In this study, we evaluated the possible interaction between miR-146a and one of its putative targets ribonuclease L (RNASEL) in the risk of sporadic melanoma. Polymorphisms rs2910164 in miR-146a and rs486907 in the RNASEL gene have both independently been associated with the risk of different cancers, and an interaction between them has been observed in nonmelanoma skin cancer. Polymorphisms rs2910164 G/C and rs486907 A/G were genotyped by restriction fragment length polymorphism analysis in 304 sporadic melanoma patients and 314 control individuals. Genotype distribution between cases and controls for each of the two polymorphisms was compared using Fisher's exact test. Epistasis between the two polymorphisms was tested by a logistic regression model. In the present study, we observed a sex-specific effect of the miR-146a rs2910164 C allele restricted to individuals carrying the RNASEL rs486907 A allele as well. Men carrying this allelic combination have the highest risk of melanoma, whereas it seems to have no effect or even an opposite relationship to melanoma risk in the female population. The results reported in the present study suggest a sex-specific interaction between miR-146a and RNASEL genes in melanoma skin cancer susceptibility, and could account for possible discordant results in association studies when stratification according to sex is not performed.

Comparison of multiple techniques for endobronchial ultrasound-transbronchial needle aspiration specimen preparation in a single institution experience.

BACKGROUND: The optimal method for specimen preparation of endobronchial ultrasound-transbronchial needle aspiration (EBUS-TBNA) is still controversial. This study aims to compare several techniques available for EBUS-TBNA specimen acquisition and processing, in order to identify the best performing technique. METHODS: We retrospectively reviewed the data of 199 consecutive patients [male, 73%; median age, 64 years (IQR: 52-74 years)] undergoing EBUS-TBNA at our institution from 2012 through 2014 for diagnosis of hilar-mediastinal lymph node enlargement suspect of neoplastic (n=139) or granulomatous (n=60) disease. All procedures were performed by two experienced bronchoscopists, under conscious sedation and local anaesthesia, using 21/22-Gauge (G) needle, without rapid on-site evaluation (ROSE). Five specimen-processing techniques were used: cytology slides in 42 cases (21%); cell-block in 25 (13%); core-tissue in 60 (30%); combination of cytology slides and core-tissue in 51 (26%); combination of cytology slides and cell-block in 21 (10%). To assess the diagnostic accuracy of each tissue-processing technique we compared the EBUS-TBNA results to those obtained with surgical lymphadenectomy, or 1-year follow-up in non-operated patients. RESULTS: Diagnostic yield, accuracy and area under the curve (AUC) were as follows. Cytology slides: 81%, 80%, 0.90; cell-block: 48%, 33%, 0.67; core-tissue: 87%, 99%, 0.96; cytology slides + core-tissue: 80%, 100%, 1.00; cytology slides + cell-block: 86%, 100%, 1.00. Cytology slides and core-tissue method showed non-significantly different diagnostic yield (P=0.435) and AUC (P=0.152). CONCLUSIONS: In our single-institution experience, cytology slides and core-tissue preparations demonstrated high and similar diagnostic performance. Cytology slides combination with core-tissue or cell-block showed the highest performance, however these combination methods were more resource-consuming.

A comparison between quantitative PCR and droplet digital PCR technologies for circulating microRNA quantification in human lung cancer.

BACKGROUND: Selected microRNAs (miRNAs) that are abnormally expressed in the serum of patients with lung cancer have recently been proposed as biomarkers of this disease. The measurement of circulating miRNAs, however, requires a highly reliable quantification method. Quantitative real-time PCR (qPCR) is the most commonly used method, but it lacks reliable endogenous reference miRNAs for normalization of results in biofluids. When used in absolute quantification, it must rely on the use of external calibrators. Droplet digital PCR (ddPCR) is a recently introduced technology that overcomes the normalization issue and may facilitate miRNA measurement. Here we compared the performance of absolute qPCR and ddPCR techniques for quantifying selected miRNAs in the serum. RESULTS: In the first experiment, three miRNAs, proposed in the literature as lung cancer biomarkers (miR-21, miR-126 and let-7a), were analyzed in a set of 15 human serum samples. Four independent qPCR and four independent ddPCR amplifications were done on the same samples and used to estimate the precision and correlation of miRNA measurements obtained with the two techniques. The precision of the two methods was evaluated by calculating the Coefficient of Variation (CV) of the four independent measurements obtained with each technique. The CV was similar or smaller in ddPCR than in qPCR for all miRNAs tested, and was significantly smaller for let-7a (p = 0.028). Linear regression analysis of the miRNA values obtained with qPCR and ddPCR showed strong correlation (p < 0.001). To validate the correlation obtained with the two techniques in the first experiment, in a second experiment the same miRNAs were measured in a larger cohort (70 human serum samples) by both qPCR and ddPCR. The correlation of miRNA analyses with the two methods was significant for all three miRNAs. Moreover, in our experiments the ddPCR technique had higher throughput than qPCR, at a similar cost-per-sample. CONCLUSIONS: Analyses of serum miRNAs performed with qPCR and ddPCR were largely concordant. Both qPCR and ddPCR can reliably be used to quantify circulating miRNAs, however, ddPCR revealed similar or greater precision and higher throughput of analysis.

Is chest X-ray screening for lung cancer in smokers cost-effective? Evidence from a population-based study in Italy.

BACKGROUND: After implementation of the PREDICA annual chest X-ray (CXR) screening program in smokers in the general practice setting of Varese-Italy a significant reduction in lung cancer-specific mortality (18 %) was observed. The objective of this study covering July 1997 through December 2006 was to estimate the cost-effectiveness of this intervention. METHODS: We examined detailed information on lung cancer (LC) cases that occurred among smokers invited to be screened in the PREDICA study (Invitation-to-screening Group, n = 5815 subjects) to estimate costs and quality-adjusted life-years (QALYs) from LC diagnosis until death. The control group consisted of 156 screening-eligible smokers from the same area, uninvited and unscreened, who developed LC and were treated by usual care. We calculated the incremental net monetary benefit (INMB) by comparing LC management in screening participants (n = 1244 subjects) and in the Invitation-to-screening group versus control group. RESULTS: The average number of QALYs since LC diagnosis was 1.7, 1.49 and 1.07, respectively, in screening participants, the invitation-to-screening group, and the control group. The average total cost (screening + management) per LC case was higher in screening participants (€17,516) and the Invitation-to-screening Group (€16,167) than in the control group (€15,503). Assuming a maximum willingness to pay of €30,000/QALY, we found that the intervention was cost-effective with high probability: 79 % for screening participation (screening participants vs. control group) and 95 % for invitation-to-screening (invitation-to-screening group vs. control group). CONCLUSIONS: Based on the PREDICA study, annual CXR screening of high-risk smokers in a general practice setting has high probability of being cost-effective with a maximum willingness to pay of €30,000/QALY.

How has peritoneal dialysis changed over the last 30 years: experience of the Verona dialysis center.

BACKGROUND: The last decade has witnessed considerable improvement in dialysis technology and changes in clinical management of patients in peritoneal dialysis (PD) with a significant impact on long term clinical outcomes. However, the identification of factors involved in this process is still not complete. METHODS: Therefore, to assess this objective, we retrospectively analyzed clinical records of 260 adult patients who started PD treatment from 1983 to 2012 in our renal unit. For the analysis, we divided them into three groups according to the time of starting dialysis: GROUP A (n: 62, 1983-1992), GROUP B (n: 66, 1993-2002) and GROUP C (n: 132, 2003 to 2012). RESULTS: Statistical analysis revealed that patients included in the GROUP C showed a reduction in mean patients' age (p = 0.03), smoking habit (p = 0.001), mean systolic blood pressure (p < 0.0001) and an increment in hemoglobin levels (p < 0.0001) and residual diuresis (p = 0.016) compared to the other two study groups. Additionally, patients included in GROUP C, mainly treated with automated peritoneal dialysis, showed a reduced risk of all-causes mortality and a decreased risk to develop acute myocardial infarction and cerebrovascular disease. Patients' age, diabetes mellitus and smoking habit were all positively associated with a significant increased risk of mortality in our PD patients, while serum albumin levels and residual diuresis were negatively correlated. CONCLUSIONS: Therefore, the present study, revealed that in the last decade there has been a growth of our PD program with a concomitant modification of our patients' characteristics. These changes, together with the evident technical advances, have caused a significant improvement of patients' survival and a decrement of the rate of hospitalization. Moreover, it reveals that our pre-dialysis care, modifying the above-mentioned factors, has been a major cause of these clinical improvements.