Cut-off Definition Research Articles

Laboratory performance of sweat conductivity for the screening of cystic fibrosis.

There are several complementary English-language guidelines for the performance of the sweat chloride test. These guidelines also incorporate information for the collection of conductivity samples. However, recommendations for the measurement and reporting of sweat conductivity are less clear than for sweat chloride. The aim of the study was to develop an understanding of the testing and reporting practices of sweat conductivity in Australasian laboratories. A survey specifically directed at conductivity testing was sent to the 12 laboratories registered with the Royal College of Pathologists of Australasia Quality Assurance Programs. Nine (75%) laboratories participated in the survey, seven of whom used Wescor Macroduct® for collecting sweat and the Wescor SWEAT·CHEK™ for conductivity testing, and the remaining two used the Wescor Nanoduct®. There was considerable variation in frequency and staffing for this test. Likewise, criteria about which patients it was inappropriate to test, definitions of adequate collection sweat rate, cutoffs and actions recommended on the basis of the result showed variations between laboratories. Variations in sweat conductivity testing and reporting reflect many of the same issues that were revealed in sweat chloride test audits and have the potential to lead to uncertainty about the result and the proper action in response to the result. We recommend that sweat testing guidelines should include clearer statements about the use of sweat conductivity.

Definition of new cut-offs of BMI and waist circumference based on body composition and insulin resistance: differences between children, adolescents and adults.

SummaryObjectiveThis study aims to determine associations between anthropometric traits, regional fat depots and insulin resistance in children, adolescents and adults to define new cut‐offs of body mass index (BMI) or waist circumference (WC).DesignCross‐sectional data were assessed in 433 children, adolescents and adults (aged: 6–60 years, BMI: 23.6 [21.0–27.7] kg m−2). Total adipose tissue (TAT), regional subcutaneous adipose tissue (SATtotal, SATtrunk) and visceral adipose tissue (VAT) were determined by whole‐body magnetic resonance imaging, fat mass by air‐displacement plethysmography. Insulin resistance was evaluated by homeostasis model assessment of insulin resistance (HOMA‐IR). Bivariate as well as partial correlations and regression analyses were used. Cut‐off values of BMI and WC related to regional fat depots and HOMA‐IR were analysed by receiver operating characteristics curve.ResultsIn adults, TAT, SATtotal and SATtrunk increased linearly with increasing BMI and WC, whereas they followed a cubic function in children and adolescents with a steep increase at BMI and WC ≥1 standard deviation score and VAT at WC ≥2 standard deviation score. Sex differences were apparent in adults with women having higher masses of TAT and SAT and men having higher VAT. Using established BMI or WC cut‐offs, correspondent masses of TAT, SATtotal, SATtrunk and VAT increased from childhood to adulthood. In all age groups, there were positive associations between BMI, WC, SATtrunk, VAT and HOMA‐IR. When compared with normative cut‐offs of BMI or WC, HOMA‐IR‐derived cut‐offs of regional fat depots were lower in all age groups.ConclusionsAssociations between BMI, WC and regional fat depots varied between children, adolescents, young and older adults. When compared with BMI‐derived and WC‐derived values, an insulin resistance‐derived cut‐off corresponded to lower masses of regional fat depots. Thus, established BMI and WC cut‐offs are not appropriate to assess metabolic disturbances associated with obesity; therefore, new cut‐offs of BMI and WC are needed for clinical practice.

Aldosterone breakthrough in dogs with naturally occurring myxomatous mitral valve disease

IntroductionAldosterone breakthrough (ABT) is the condition in which angiotensin converting enzyme inhibitors (ACEIs) and/or angiotensin receptor blockers fail to effectively suppress the activity of the renin angiotensin aldosterone system. The objective of this study was to determine if ABT occurs in dogs with naturally occurring myxomatous mitral valve disease receiving an ACEI, using the urine aldosterone to creatinine ratio (UAldo:C) as a measure of renin angiotensin aldosterone system activation. Animals, Materials and MethodsThis study includes 39 dogs with myxomatous mitral valve disease. A UAldo:C cut-off definition (derived from a normal population of healthy, adult, and client-owned dogs) was used to determine the prevalence of ABT in this population. Spearman analysis and univariate logistic regression were used to evaluate the relationship between UAldo:C and ABT (yes/no) and eight variables (age, serum K+ concentration, serum creatinine concentration, ACEI therapy duration and ACEI dosage, furosemide therapy duration and furosemide dosage, and urine sample storage time). Finally, the UAldo:C in dogs receiving spironolactone, as part congestive heart failure (CHF) therapy, was compared to dogs with CHF that were not receiving spironolactone. ResultsThe prevalence of ABT was 32% in dogs with CHF and 30% in dogs without CHF. There was no relationship between either the UAldo:C or the likelihood of ABT and the eight variables. Therapy with spironolactone lead to a significant elevation of the UAldo:C. DiscussionUsing the UAldo:C and a relatively stringent definition of ABT, it appears that incomplete RAAS blockade is common in dogs with MMVD receiving an ACEI. The prevalence of ABT in this canine population mirrors that reported in humans. While the mechanism of ABT is likely multifactorial and still poorly understood, the proven existence of ABT in dogs offers the potential to improve the prognosis for MMVD with the addition of a mineralocorticoid receptor blocker to current therapeutic regimens. ConclusionsApproximately 30% of dogs being treated for heart disease and CHF satisfied the definition of ABT. Identifying patient subpopulations experiencing ABT may help guide future study design and clinical decision-making.

Hyperglycemia and survival in solid tumors: A systematic review and meta-analysis.

e18158 Background: Diabetes is associated with adverse cancer outcomes. However, the effect of hyperglycemia independent of diabetes is unclear. Here we report on a meta-analysis exploring the effect of hyperglycemia on outcomes of solid tumors, and the influence of clinical factors on this association. Methods: A systematic search of electronic databases identified publications exploring the effect of hyperglycemia on overall (OS), disease-free (DFS) or progression-free survival (PFS). Definitions of hyperglycemia (fasting blood glucose, random blood glucose or HbA1c) and cut-offs varied between studies. Data from studies reporting a hazard ratio (HR) and 95% confidence interval (CI) or a p -value were pooled in a meta-analysis using generic inverse-variance and random effects modeling. Subgroup analyses were conducted based on method of hyperglycemia measurement (HbA1c, other) and tumor stage (early, advanced, mixed). Meta-regression was performed to evaluate the influence of clinical characteristics including baseline diabetes status on the HR for OS. All statistical tests were two-sided. Results: Eight studies comprising a total of 4342 patients were included. All studies reported HRs for OS. Two studies reported DFS outcomes, and two reported PFS. Hyperglycemia was associated with worse OS (HR 2.07, 95% CI = 1.70 - 2.52; P < 0.001) and DFS (HR 1.61, 95% CI = 1.04 - 2.49; P < 0.001), but did not decrease PFS (HR 1.08, 95% CI = 0.72 - 1.62; P = 0.71). The association with worse OS maintained in subgroups based on method of hyperglycemia measurement (subgroup difference P = 0.65) and tumor stage ( P= 0.47). Meta-regression analyses did not identify any factors significantly altering the magnitude of association between hyperglycemia and OS (see Table). Conclusions: Hyperglycemia is associated with adverse OS and DFS in patients with cancer, and the therapeutic role of optimal glycemic control warrants further investigation. [Table: see text]

Late-night salivary cortisol: cut-off definition and diagnostic value in Cushing's syndrome

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Assessment of Adherence to the STARD Statement for the Quality of Reports on Diagnostic Studies Published on the Iranian Journal of Radiology

Background: Standards for the Reporting of diagnostic accuracy studies (STARD) statement was developed to improve the accuracy and completeness of reporting of studies of diagnostic accuracy. It allows readers evaluate the internal and external validity and enable readers to assess the potential for bias in the study and the generalizability of the results. Objectives: Up to date, few data exist about the quality of diagnostic accuracy studies in Iran. Therefore, we evaluated the extent of adherence to the STARD for the quality of reports on diagnostic accuracy studies in the Iranian journal of radiology. Methods: In this cross-sectional descriptive study, we searched Iranian journal of radiology and retrieved all diagnostic accuracy studies for recent 3 years and evaluated their adherence to STARD. We used STARD checklist (version January 2003) available at www.stard-statement.org Results: We identified about 14 potential diagnostic accuracy studies meeting our inclusion criteria. In this preliminary study 50% identified the article as diagnostic accuracy study in the title or abstract (MeSH heading sensitivity and specificity). 4% reported details of blinding. 3% had a flow diagram. 38% described the number, training and expertise of the persons reading the index tests and the reference standard. 25% reported definition of cutoffs and/or categories of the results of the index tests and the reference standard. Only 40% of the reports of diagnostic evaluations reported precision for the estimates of diagnostic accuracy. The occurrence of uninterpretable, indeterminate and intermediate test results were 30%. The report time interval from the index tests to the reference standard was 40%. Only 40% of articles reported on more than 50% of STARD items, while no articles reported on more than 85%. A flow chart was presented in 3 articles. Assessment of reporting on individual items of the STARD statement revealed wide variation, with some items described in 15% of articles and others in 100%. Mean STARD score (0 - 25 points available) was 14.8 (range, 5.5 19.5). Conclusions: These findings show non-adherence to the STARD. The authors, reviewers, and editors should pay more attention to reporting by checking STARD statement items and including a flow diagram to represent study design and patient flow.

The balance between the serum levels of IL-6 and IL-10 cytokines discriminates mild and severe acute pneumonia.

BackgroundTo identify markers for earlier diagnosis of severe pneumonia, we assess the correlation between serum cytokine profile of children with different pneumonia severity.MethodsIn 25 hospitalized children, 7 with mild pneumonia and 18 with severe pneumonia, the serum concentration of 11 cytokines in three sampling times were dosed. Statistical analysis included parametric and non-parametric tests, Pearson correlation and ROC curve for cut-off definition of cytokines.ResultsAt admission, IL-6 serum levels were high in mild or severe pneumonia, and was associated to vomiting (P = 0.019) in both groups; and also to dyspnea (P = 0.012) and white blood cell count (P = 0.045) in patients with severe pneumonia. IL-10 levels were also high in patients with pneumonia and were associated to lymphocytosis (P = 0.025). The ROC curve of the IL-6:IL-10 serum levels ratio discriminated severe pneumonia cases at admission, and persistence of infection in the third day of antibiotic therapy, with positive predictive values of 93% and 89%, respectively.ConclusionsThe balance between IL-6 and IL-10 serum levels showed to be a more discriminative marker for severity definition and evaluation of recovery in patients with pneumonia.Electronic supplementary materialThe online version of this article (doi:10.1186/s12890-016-0324-z) contains supplementary material, which is available to authorized users.

Basophil Reactivity as Biomarker in Immediate Drug Hypersensitivity Reactions-Potential and Limitations.

Immediate drug hypersensitivity reactions (DHRs) resemble typical immunoglobulin E (IgE)-mediated symptoms. Clinical manifestations range from local skin reactions, gastrointestinal and/or respiratory symptoms to severe systemic involvement with potential fatal outcome. Depending on the substance group of the eliciting drug the correct diagnosis is a major challenge. Skin testing and in vitro diagnostics are often unreliable and not reproducible. The involvement of drug-specific IgE is questionable in many cases. The culprit substance (parent drug or metabolite) and potential cross-reacting compounds are difficult to identify, patient history and drug provocation testing often remain the only means for diagnosis. Hence, several groups proposed basophil activation test (BAT) for the diagnosis of immediate DHRs as basophils are well-known effector cells in allergic reactions. However, the usefulness of BAT in immediate DHRs is highly variable and dependent on the drug itself plus its capacity to spontaneously conjugate to serum proteins. Stimulation with pure solutions of the parent drug or metabolites thereof vs. drug-protein conjugates may influence sensitivity and specificity of the test. We thus, reviewed the available literature about the use of BAT for diagnosing immediate DHRs against drug classes such as antibiotics, radio contrast media, neuromuscular blocking agents, non-steroidal anti-inflammatory drugs, and biologicals. Influencing factors like the selection of stimulants or of the identification and activation markers, the stimulation protocol, gating strategies, and cut-off definition are addressed in this overview on BAT performance. The overall aim is to evaluate the suitability of BAT as biomarker for the diagnosis of immediate drug-induced hypersensitivity reactions.

Hotspot identification in the clothing industry using social life cycle assessment—opportunities and challenges of input-output modelling

A cradle-to-gate, input/output-based social life cycle assessment (SLCA) was conducted using the Swedish clothing consumption as a case study. The aim was to investigate the influence of the cut-off rule and the definition of "hotspots" in social hotspot assessment. A second aim was to identify social hotspots of Swedish clothing on a national level. The case study was based on the SLCA methodology provided in the Guidelines for Social Life Cycle Assessment of Products (BenoIt and Mazijn 2009). An input/output model was used to define the product system from cradle to gate. The negative social hotspots were evaluated for a set of social indicators that were selected by consumers. The impact assessment was conducted on a sector and country level by using the Social Hotspots Database. The identified sectors of the economy with high and very high levels of risk were listed for each social indicator. The results pinpointed some hotspots throughout the supply chain for Swedish clothing consumption. Some unexpected sectors such as commerce and business services in Bangladesh were identified as important hotspots as well as main sectors in the production phase such as plant fibres, textiles and garments that would be expected also on the bases of a traditional process analysis. A sensitivity analysis on different cut-off values showed the extent to which the choice of cut-off rule can directly affect the results via influence over the number of country-specific sectors (CSSs) in the product system. The influence of the hotspot definition was investigated by evaluating the working hour intensity for low- and medium-risk levels for three different indicators. The results show that for child labour, 92 % of the share of working hours was associated with low- and medium-risk levels. Therefore, the evaluation of risk levels other than high and very high can provide a more complete picture of the hotspots. The application of input/output-based SLCA on the clothing production supply chain provided a more complete picture of the social hotspots than with traditional process-based SLCA. Some unexpected sectors related to commerce and business appeared as social hotspots in the clothing industry. The study explored some important parameters in applying an input/output-based SLCA. The results show that the cut-off values and definition of hotspots in relation to risk levels can directly influence the results.

Sensitivity Analysis of the MGMT-STP27 Model and Impact of Genetic and Epigenetic Context to Predict the MGMT Methylation Status in Gliomas and Other Tumors

The methylation status of the O(6)-methylguanine-DNA methyltransferase (MGMT) gene is an important predictive biomarker for benefit from alkylating agent therapy in glioblastoma. Our model MGMT-STP27 allows prediction of the methylation status of the MGMT promoter using data from the Illumina's Human Methylation BeadChips (HM-27K and HM-450K) that is publically available for many cancer data sets. Here, we investigate the impact of the context of genetic and epigenetic alterations and tumor type on the classification and report on technical aspects, such as robustness of cutoff definition and preprocessing of the data. The association between gene copy number variation, predicted MGMT methylation, and MGMT expression revealed agene dosage effect on MGMT expression in lower grade glioma (World Health Organization gradeII/III) that in contrast to glioblastoma usually carry two copies of chromosome 10 on which MGMT resides (10q26.3). This implies some MGMT expression, potentially conferring residual repair function blunting the therapeutic effect of alkylating agents. A sensitivity analyses corroborated the performance of the original cutoff for various optimization criteria and for most data preprocessing methods. Finally, we propose an R package mgmtstp27 that allows prediction of the methylation status of the MGMT promoter and calculation of appropriate confidence and/or prediction intervals. Overall, MGMT-STP27 is a robust model for MGMT classification that is independent of tumor type and is adapted for single sample prediction.

The "ABC" of Virus-Specific T Cell Immunity in Solid Organ Transplantation.

Transplant patients are at increased risk of viral complications due to impaired control of viral replication, resulting from HLA mismatching between graft and host and the immunosuppression needed to avert alloimmune reactions. In the past decade, quantitative viral load measurements have become widely available to identify patients at risk and to inform treatment decisions with respect to immunosuppressive drugs and antiviral therapies. Because viral loads are viewed as the result of viral replication and virus-specific immune control, virus-specific T cell monitoring has been explored to optimize management of adenovirus, BK polyomavirus and cytomegalovirus ("ABC") in transplant patients. Although most studies are descriptive using different technologies, the overall results show that the quantity and quality of virus-specific T cells inversely correlate with viral replication, whereby strong cellular immune responses are associated with containment of viral replication. The key obstacles to the introduction of assays for virus-specific T cells into clinical practice is the definition of reliable cutoffs for clinical decision making, the poor negative predictive value of some assays, and the absence of interventional trials justifying changes of antiviral treatment or immunosuppression. More clinical research is needed using optimized assays and targets before standardization and commutability can be envisaged as achieved for viral load testing.

Performance of 14 rubella IgG immunoassays on samples with low positive or negative haemagglutination inhibition results

BackgroundRubella IgG testing is routinely done in prenatal care and seroepidemiological studies. Recently concern was raised that seropositivity rates were decreasing questioning vaccination policies. Manufacturers of rubella IgG assays and authors of seroepidemiological studies use different cut-offs for the definition of seropositivity. As rubella virus circulation is reduced since many years, seronegativity rates might be overestimated using an inappropriate cut-off. ObjectivesUsing different cut-off definitions we compared fourteen current rubella IgG immunoassays for sensitivity and qualitative result concordance in samples with low positive or negative haemagglutination inhibition (HI) titre. Study design150 clinical samples from patients and health care workers were included in the study. All samples were measured in 14 different rubella IgG immunoassays. Seropositivity was defined using recombinant rubella IgG immunoblot as reference standard. ResultsThe concordance of qualitative results using the manufacturers cut-off definitions was 56.4% if grey-zone results were analysed separately and 69.8% if grey-zone results were defined as positive. Using universal cut-offs of 10IU/ml or 15IU/ml the concordance was 70% and 61.4% respectively. Using the different cut-off definitions up to 71 out of the 124 immunoblot-positive samples tested negative in the immunoassays. The mean coefficient of variation (CV) of quantitative results in positive samples was 51% (range 19–113%). ConclusionsDetermination of rubella immunity by measurement of rubella-IgG in a population with high vaccination coverage with current assays leads to a high number of false negative results. The value of routine rubella antibody testing in countries with high vaccination coverage should be discussed.

Interlaboratory variability of MIB1 staining in well-differentiated pancreatic neuroendocrine tumors.

Neuroendocrine tumors (NET) are routinely graded and staged to judge prognosis. Proliferation index using MIB1 staining has been introduced to assess grading. There are vivid discussions on cutoff definitions, automated counting, and interobserver variability. However, no data exist regarding interlaboratory reproducibility for low proliferation indices which are of importance to discriminate between G1 and G2 NET. We performed MIB1 staining in three different university hospital-based pathology laboratories on a tissue micro array (TMA) of a well-characterized patient cohort, containing pancreatic NET of 61 patients. To calculate the proliferation index, number of positive tumor nuclei was divided by the total number of tumor nuclei. Labeling index was compared to mitotic counts in whole tissue sections and to clinical outcome. Linear regression analysis, intraclass comparison, and log-rank analysis were performed. Intraclass correlation showed moderate-to-fair agreement. Especially low proliferating tumors were affected by interlaboratory differences. Log-rank analysis was performed for each lab and resulted in three different cutoffs (5.0, 3.0, and 0.5 %). Every calculated cutoff stratified the patient cohort to a significant extent for the underlying stain (p < 0.001, <0.001, and <0.001) but showed no or lesser significance when applied to the other stains. Significant and relevant interlab differences for MIB1 exist. Since the MIB1 proliferation index influences grading, local cutoffs or external standardization should urgently be introduced to achieve reliability and reproducibility.

Solvation station: microsolvation for modeling vibrational sum-frequency spectra of acids at aqueous interfaces.

Vibrational sum-frequency spectra of a pair of poly(methacrylic acid) isomers at an oil/water interface and glutaric acid at an air/water interface were calculated in the carbonyl stretching region. Orientational, conformational, and solvation information was determined using classical molecular dynamics (MD), while second-order susceptibility vibrational response tensors were determined for a set of density functional theory (DFT) structures. The DFT structures were microsolvated with water molecules corresponding to the major solvation states present in the MD calculations. The inclusion of the microsolvating waters incorporates solvation effects important to the carboxylic acid stretching modes in the studied spectral region. The calculated spectra strongly agree with experimental spectra when a cutoff of 1.975 Å is used to define a hydrogen bond in the MD trajectories. With the chosen cutoff, the most common solvation state of the carboxylic acid moieties involves a single hydrogen bond to the carbonyl oxygen and a single hydrogen bond to the carboxylic acid hydrogen. The sensitivity of the spectra to the hydrogen bond cutoff definition and the included DFT structures was investigated. Moderate changes in the relative intensities of the contributing peaks were found in both cases. Shortening the hydrogen bond cutoff definition predictably leads to a decrease in the relative intensity of peaks corresponding to well-solvated structures, while altering the set of DFT solvation structures results in more complex behavior that is dependent on the specific structures included.

Clinical Neuropathology practice guide 5-2015: MGMT methylation pyrosequencing in glioblastoma: unresolved issues and open questions.

O6-methylguanine-methyltransferase (MGMT) promoter methylation status has prognostic and, in the subpopulation of elderly patients, predictive value in newly diagnosed glioblastoma. Therefore, knowledge of the MGMT promoter methylation status is important for clinical decision-making. So far, MGMT testing has been limited by the lack of a robust test with sufficiently high analytical performance. Recently, one of several available pyrosequencing protocols has been shown to be an accurate and robust method for MGMT testing in an intra- and interlaboratory ring trial. However, some uncertainties remain with regard to methodological issues, cut-off definitions, and optimal use in the clinical setting. In this article, we highlight and discuss several of these open questions. The main unresolved issues are the definition of the most relevant CpG sites to analyze for clinical purposes and the determination of a cut-off value for dichotomization of quantitative MGMT pyrosequencing results into “MGMT methylated” and “MGMT unmethylated” patient subgroups as a basis for further treatment decisions.

Research progress on the clinical value of Ki-67 in breast cancer and its cut-off definition

Ki-67 has an important application value in clinical practice. However, it is still a little tough in clinical application because of the debate on the cut-off definition of Ki-67 index. This review summarizes most studies on the prognostic and predictive value of Ki-67, analyzes the reasons for the discrepancies among the studies cited, and presents the necessity and clinical significance of scientifically defining the cut-off of Ki-67 index, providing a theoretical basis for Ki-67 in clinical application.

Prognostic value of abnormally expressed long non-coding RNAs in patients with osteosarcoma: a meta-analysis

// Fashuai Wu 1, * , Deyao Shi 1, * , Shidai Mu 2 , Yu Huang 3 , Feng Gao 1 , Xiangcheng Qing 1 and Zengwu Shao 1 1 Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China 2 Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China 3 Department of Otorhinolaryngology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China * These authors contributed equally to this work Correspondence to: Zengwu Shao, email: profszw1962@163.com Keywords: lncRNA; osteosarcoma; prognosis; biomarker; meta-analysis Received: May 23, 2017 Accepted: December 26, 2017 Published: January 12, 2018 ABSTRACT Long non-coding RNAs (lncRNAs) contribute to progression of various cancers including osteosarcoma through abnormal regulation of cancer-related cellular processes. Recent researches have shown that various lncRNAs are abnormally expressed in osteosarcoma and associated with the prognosis. With the aim of gaining a better insight into the association between expression level of lncRNAs and prognosis of osteosarcoma, multiple databases including PubMed, Embase, Cochrane Library and Web of Science were carefully searched for available studies up to March 14, 2017. Finally, 19 publications with 1298 patients matched our inclusion criteria and were evaluated in our meta-analysis. The quality of each study was scored using the Newcastle-Ottawa Scale and studies not reaching a minimum threshold were excluded. Results of the meta-analysis demonstrated that abnormal expression level of lncRNAs predicted poor overall survival (pooled HR 3.064, 95% CI: 2.487–3.775) and event-free survival (pooled HR 2.642, 95% CI 1.759–3.970) in osteosarcoma and subgroup analysis showed consistent prognostic value. Furthermore, combining data of Cox multivariable analysis indicated that abnormal expression level of lncRNAs was an independent prognostic marker for overall survival (pooled HR 2.864, 95% CI: 2.246–3.651) in osteosarcoma patients. The clinicopathological parameters analysis further showed that abnormal expression level of lncRNAs was correlated with tumor size, tumor stage, metastasis and differentiation grade of osteosarcoma. Limitations of the meta-analysis included variation of cut-off value definition, Chinese provenance of most studies, publication bias and so on. In conclusion, this meta-analysis suggested that abnormal expression level of lncRNAs has a promising future for predicting the prognosis of patients with osteosarcoma.

Unpredictable susceptibility of emerging clinical moulds to tri-azoles: review of the literature and upcoming challenges for mould identification.

Tri-azoles represent the front-line drugs for the treatment of mould diseases; nevertheless, some emerging moulds, such as Fusarium spp., Scedosporium spp., Mucorales and others, may be less susceptible or resistant to these antifungals. A review of the literature was conducted on the susceptibility of rare moulds to the tri-azoles itraconazole, posaconazole and voriconazole. Particular attention was paid to isolates identified by molecular analyses. The range of susceptibility values described for the three tri-azoles was frequently large (from 0.06 to >16), and a high variability was found within each species; isolates were rarely reported as entirely susceptible to all tri-azoles. In addition, the susceptibility of 76 emerging moulds from our collection (including Hypocreales, Dothideomycetes, Scedosporium spp., Mucorales and rare Aspergillus spp.) to itraconazole and voriconazole was determined by the Clinical and Laboratory Standards Institute (CLSI) M38-A2 and European Committee for Antimicrobial Susceptibility Testing (EUCAST) methods. Susceptibility discrepancies (of two dilutions) were found comparing CLSI and EUCAST for Dothideomycetes; the values for the remaining moulds were similar. More practical, faster and inexpensive susceptibility tools are welcome for testing emerging moulds, as these tests still represent a critical tool to support clinicians on the selection of proper antifungal treatment. The susceptibility of emerging moulds to tri-azoles cannot be predicted exclusively following mould identification, as the isolates' susceptibilities showed highly variable values. Some emerging moulds still remain very difficult to identity, even following standard molecular analyses which result in complex fungal collections. This fact limits the definition of epidemiological cut-offs and clinical breakpoints that are still imperative for emerging moulds.

Flexible lab-tailored cut-offs for suitability of formalin-fixed tumor samples for diagnostic mutational analyses.

The selection of proper tissues from formalin-fixed and paraffin-embedded tumors before diagnostic molecular testing is responsibility of the pathologist and represents a crucial step to produce reliable test results. The international guidelines suggest two cut-offs, one for the percentage and one for the number of tumor cells, in order to enrich the tumor content before DNA extraction. The aim of the present work was two-fold: to evaluate to what extent a low percentage or absolute number of tumor cells can be qualified for somatic mutation testing; and to determine how assay sensitivities can guide pathologists towards a better definition of morphology-based adequacy cut-offs. We tested 1797 tumor specimens from melanomas, colorectal and lung adenocarcinomas. Respectively, their BRAF, K-RAS and EGFR genes were analyzed at specific exons by mutation-enriched PCR, pyrosequencing, direct sequencing and real-time PCR methods. We demonstrate that poorly cellular specimens do not modify the frequency distribution of either mutated or wild-type DNA samples nor that of specific mutations. This observation suggests that currently recommended cut-offs for adequacy of specimens to be processed for molecular assays seem to be too much stringent in a laboratory context that performs highly sensitive routine analytical methods. In conclusion, new cut-offs are needed based on test sensitivities and documented tumor heterogeneity.

C-reactive protein (CRP), interferon gamma-inducible protein 10 (IP-10), and lipopolysaccharide (LPS) are associated with risk of tuberculosis after initiation of antiretroviral therapy in resource-limited settings.

ObjectiveThe association between pre-antiretroviral (ART) inflammation and immune activation and risk for incident tuberculosis (TB) after ART initiation among adults is uncertain.DesignNested case-control study (n = 332) within ACTG PEARLS trial of three ART regimens among 1571 HIV-infected, treatment-naïve adults in 9 countries. We compared cases (participants with incident TB diagnosed by 96 weeks) to a random sample of controls (participants who did not develop TB, stratified by country and treatment arm).MethodsWe measured pre-ART C-reactive protein (CRP), EndoCab IgM, ferritin, interferon gamma (IFN-γ), interleukin 6 (IL-6), interferon gamma-inducible protein 10 (IP-10), lipopolysaccharide (LPS), soluble CD14 (sCD14), tumor necrosis factor alpha (TNF-α), and CD4/DR+/38+ and CD8/DR+/38+ T cells. Markers were defined according to established cutoff definitions when available, 75th percentile of measured values when not, and detectable versus undetectable for LPS. Using logistic regression, we measured associations between biomarkers and incident TB, adjusting for age, sex, study site, treatment arm, baseline CD4 and log10 viral load. We assessed the discriminatory value of biomarkers using receiver operating characteristic (ROC) analysis.ResultsSeventy-seven persons (4.9%) developed incident TB during follow-up. Elevated baseline CRP (aOR 3.25, 95% CI: 1.55–6.81) and IP-10 (aOR 1.89, 95% CI: 1.05–3.39), detectable plasma LPS (aOR 2.39, 95% CI: 1.13–5.06), and the established TB risk factors anemia and hypoalbuminemia were independently associated with incident TB. In ROC analysis, CRP, albumin, and LPS improved discrimination only modestly for TB risk when added to baseline routine patient characteristics including CD4 count, body mass index, and prior TB.ConclusionIncident TB occurs commonly after ART initiation. Although associated with higher post-ART TB risk, baseline CRP, IP-10, and LPS add limited value to routine patient characteristics in discriminating who develops active TB. Besides determining ideal cutoffs for these biomarkers, additional biomarkers should be sought that predict TB disease in ART initiators.