Conventional Serum Markers Research Articles

Synthesis, Characterization, and Potential Usefulness in Liver Function Assessment of Novel Bile Acid Derivatives with Near-Infrared Fluorescence (NIRBAD).

Conventional serum markers often fail to accurately detect cholestasis accompanying many liver diseases. Although elevation in serum bile acid (BA) levels sensitively reflects impaired hepatobiliary function, other factors altering BA pool size and enterohepatic circulation can affect these levels. To develop fluorescent probes for extracorporeal noninvasive hepatobiliary function assessment by real-time monitoring methods, 1,3-dipolar cycloaddition reactions were used to conjugate near-infrared (NIR) fluorochromes with azide-functionalized BA derivatives (BAD). The resulting compounds (NIRBADs) were chromatographically (FC and PTLC) purified (>95%) and characterized by fluorimetry, 1H NMR, and HRMS using ESI ionization coupled to quadrupole TOF mass analysis. Transport studies using CHO cells stably expressing the BA carrier NTCP were performed by flow cytometry. Extracorporeal fluorescence was detected in anesthetized rats by high-resolution imaging analysis. Three NIRBADs were synthesized by conjugating alkynocyanine 718 with cholic acid (CA) at the COOH group via an ester (NIRBAD-1) or amide (NIRBAD-3) spacer, or at the 3α-position by a triazole link (NIRBAD-2). NIRBADs were efficiently taken up by cells expressing NTCP, which was inhibited by taurocholic acid (TCA). Following i.v. administration of NIRBAD-3 to rats, liver uptake and consequent release of NIR fluorescence could be extracorporeally monitored. This transient organ-specific handling contrasted with the absence of release to the intestine of alkynocyanine 718 and the lack of hepatotropism observed with other probes, such as indocyanine green. NIRBAD-3 administration did not alter serum biomarkers of hepatic and renal toxicity. NIRBADs can serve as probes to evaluate hepatobiliary function by noninvasive extracorporeal methods.

Circ_0006949 as a potential non-invasive diagnosis biomarker promotes the proliferation of NSCLC cells via miR-4673/GLUL axis

The 5-year survival for non-small cell lung cancer (NSCLC) remains <20 %, primarily due to the early symptoms of lung cancer are inconspicuous. Prompt identification and medical intervention could serve as effective strategies for mitigating the death rate. We therefore set out to identify biomarkers to help diagnose NSCLC. CircRNA microarray and qRT-PCR reveal that sputum circ_0006949 is a potential biomarker for the early diagnosis and therapy of NSCLC, which can enhance the proliferation and clone formation, regulate the cell cycle, and accelerate the migration and invasion of NSCLC cells. Circ_0006949 and miR-4673 are predominantly co-localized in the cytoplasm of NSCLC cell lines and tissues; it upregulates GLUL by adsorption of miR-4673 through competing endogenous RNAs mechanism. The circ_0006949/miR-4673/GLUL axis exerts pro-cancer effects in vitro and in vivo. Circ_0006949 can boost GLUL catalytic activity, and they are highly expressed in NSCLC tissues and correlate with poor prognosis. In summary, circ_0006949 is a potential biomarker for the early diagnosis and therapy of NSCLC. This novel sputum circRNA is statistically more predictive than conventional serum markers for NSCLC diagnosis. Non-invasive detection of patients with early-stage NSCLC using sputum has shown good potential for routine diagnosis and possible screening.

Significance of Serum Leucine-rich Alpha-2 Glycoprotein as a Diagnostic Marker in Pediatric Inflammatory Bowel Disease.

Serum leucine-rich alpha-2 glycoprotein (LRG) has been utilized for adult inflammatory bowel disease (IBD); however, its efficacy in pediatric IBD remains unknown. The aim of this study was to compare the diagnostic accuracy of serum LRG for pediatric IBD with that of current inflammatory markers, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). This retrospective case-control study included pediatric patients, aged <16 years, who underwent colonoscopy and/or esophagogastroduodenoscopy between April 2017 and March 2022. All eligible patients were divided into two groups: patients with IBD, diagnosed with ulcerative colitis and Crohn's disease, and non-IBD controls. The optimal cut-off value of serum LRG for IBD diagnosis was determined from receiver operating characteristic analysis, and diagnostic accuracy of serum LRG was compared to serum ESR and CRP. A total of 53 patients (24 with IBD and 29 non-IBD controls) met the inclusion criteria. The cut-off value of serum LRG for IBD diagnosis was determined to be 19.5 μg/ml. At this cut-off value, serum LRG had a positive predictive value (PPV) of 0.80 and negative predictive value (NPV) of 0.88. In contrast, PPV and NPV were 0.78 and 0.70 for serum ESR and 0.82 and 0.72 for serum CRP, respectively. Serum LRG can be a potential diagnostic marker for pediatric IBD, with higher diagnostic accuracy than that of the conventional serum markers ESR and CRP.

The Role of urinary kidney injury molecule 1 (KIM-1) in monitoring the treatment response in Egyptian Lupus Nephritis Patients

Lupus nephritis (LN) affects almost two-thirds of systemic lupus erythematosus (SLE) patients. Despite initial aggressive therapy, up to 25% of patients with LN will progress to permanent renal damage. Conventional serum markers for LN lack the sensitivity of an ideal biomarker. Urinary kidney injury molecule-1 (UKIM-1) is an excellent biomarker for early diagnosis of acute kidney injury and predicting renal outcomes. This study intended to determine the predictive performance of UKIM-1 among LN patients in response to induction therapy by assessing and correlating its levels with renal disease activity. The study included 60 SLE patients divided into 20 SLE patients with active lupus nephritis, 20 SLE patients with inactive lupus nephritis, and 20 lupus patients without nephritis as controls. The study was completed after six months from induction treatment. UKIM-1 was measured by an enzyme linked immunosorbent assay at baseline, three-month follow-up and after complete induction therapy. At baseline, the mean serum creatinine and mean UKIM-1 were 1.7 ±0.7 mg/dL and 10.3 ±1.2 ng/dL, respectively in active LN patients. The mean UKIM-1 levels of complete response and partial response groups were 9.8 ±0.9 ng/mL and 11.3 ±1.0 ng/mL respectively. Based on receiver operating characteristics curve analysis, we found a better diagnostic performance of UKIM-1 to predict response to induction treatment, outperforming conventional biomarkers. The sensitivity and specificity were 84.6% and 85.7 %, respectively at an area under the curve of 0.896 and the best cut-off level was ≤10.6 ng/mL. In conclusion, UKIM-1 performed better than conventional biomarkers in predicting response to treatment of active LN.

Preliminary study of the interactive effects of coronary heart disease and lacunar infarction on renal function in patients with type 2 diabetes mellitus by gender

BackgroundCoronary heart disease (CHD) and lacunar infarction (LI) are the most common cardio- cerebrovascular complications of type 2 diabetes mellitus (T2DM) and a recognized risk factor for renal injury. Although a unidirectional association of CHD or LI with T2DM or the kidney has been demonstrated, however, it remains unknown whether there is an interactive effect of the coexistence of CHD and LI on renal function in T2DM patients. The aim of our study was to investigate the interaction between CHD and LI on renal function in gender-specific patients with T2DM and the association between cardio-cerebrovascular disease-related conventional serum markers and the estimated glomerular filtration rate (eGFR). MethodsWe conducted a cross-sectional study in Beijing and Tianjin from April 2019 to August 2021. Participants with T2DM aged ≥18 years were asked to complete a one-to-one questionnaire and physical examination. ResultsIn this study, 389 eligible patients with T2DM were included, with a mean age of 63.04 ± 9.41 years, of whom 200 (51.41 %) were male. The proportions of patients with CHD, LI, and both CHD and LI were 28.53 %, 24.42 %, and 11.05 %, respectively. Compared to T2DM patients without either CHD or LI, those with both CHD and LI were found to have a significantly greater risk of reduced eGFR (OR: 12.82, 95 % CI 5.06–32.52, P < 0.001) than those with CHD alone (OR: 2.42, 95 % CI 1.37–3.00, P = 0.004) or LI alone (OR: 1.15, 95 % CI 0.61–2.18, P = 0.664). The combined presence of CHD and LI is associated with a significantly greater risk of decreased eGFR in female T2DM patients compared to their male counterparts. We found both multiplicative and additive effects in all T2DM patients; however, when stratified by sex, only multiplicative effects were observed. After controlling for interference from CHD, LI, and age, we found that total cholesterol (TC) was negatively correlated with eGFR in females (r = −0.156, P = 0.034), and low-density lipoprotein cholesterol (LDL-C) was negatively correlated with eGFR in males (r = −0.229, P = 0.001). ConclusionThis study provides novel evidence that the synergistic effect of CHD and LI on renal injury in patients with T2DM is significantly greater than their individual effects. Women with T2DM who have both CHD and LI are at a 4.85-fold higher risk of decreased eGFR than men. Therefore, increased clinical attention should be given to preventing and treating vascular complications in T2DM patients, as well as aggressively reducing lipid levels, particularly TC and LDL-C, to delay or prevent renal dysfunction in T2DM patients.

A 12 Gene Signature Accurately Predicts Multiple Myeloma Progression from Monoclonal Gammopathy of Undetermined Significance

Introduction: More than 176,000 individuals are diagnosed with multiple myeloma (MM) worldwide per year. MM is always preceded by an asymptomatic phase, monoclonal gammopathy of undetermined significance (MGUS) or smoldering multiple myeloma (SMM). MGUS is found in 2.4% of individuals ≥ 50 years. The results of long-term retrospective follow-up studies have shown a 1.0% annual risk of MGUS progressing to MM. At present, using conventional serum markers, MGUS patients can be stratified into clinical risk groups. However, these models were based on retrospective analyses of cohorts tested for a limited set of clinical variables which did not incorporate genomic properties and are inconsistent as suggested by recent studies. Our previous work has established that the GEP70 signature can identify high-risk MM patients with median survival is about 2.5 years. A robust high-risk molecular signature predicting MGUS progression is still lacking. Methods: A total of 268 MGUS patients with gene expression profiles (GEPs) were included in this study. Twenty-six progressed to MM within 10 years and were considered the high-risk group, while 242 MGUS patients not progressing to MM within 10 years were referred to as the stable group. Plasma cells were enriched from bone marrow aspirates to >85% purity using anti-CD138 immunomagnetic beads. GEP was performed with the Affymetrix U133 Plus 2.0 microarray platform, using 54,613 gene probes. Affymetrix signals MAS5.0 normalized and transformed by log-base 2 for each sample. We used 3-fold cross-validation to identify the number of genes at the top of this list, probes were ranked by q-values, which collectively maximized the concordance between risk score and MGUS progression. Gene scores were computed by subtracting the average of the expression of down-regulated genes from the average of the expression of up-regulated genes. The overall accuracy of using a gene score to predict the outcome of progression was analyzed by a receiver operating characteristic curve (ROC) and its area under the curve (AUC). MGUS progression was assessed by Kaplan-Meier curves comparing the different groups. Cox proportional hazards regression analyses were applied to evaluate the significance of the gene signature for MGUS progression. Results: Of the 54,613 probes, using q statistics at <0.01, 81 probes were significantly associated with MGUS progression. After the 3-fold cross-validation analysis, the top twelve probes that appeared in each validation and maximized the concordance between risk score and MGUS progression were included in the gene signature 12 (GS12). The up-regulated genes were MTRR (5p15.31) and BAG1 (9p13.3). The down-regulated genes were ECHDC2(1p32.3), NLRP7(19q13.42), BIRC3(11q22.2), PLEKHF1(19q12), PTPN1(20q13.13), CKAP2(13q14.3), RNF5(6p21.32), LIMA1(12q13.12), CEACAM3(19q13.2) and LOC100128079(16p12.1). The ROC analysis showed that the GS12 could accurately predict the MGUS progression in all 268 MGUS patients (AUC = 0.920). An optimal cut-point for risk of progression by the GS12 score was found to be 2.73, which identified a subset of 23 (8.6%) patients with a 10-year progression probability of 73.9%. The remainder of the 245 patients (91.4%) had a probability of progression of only 3.7%. The sensitivity and specificity were 65.4% and 97.5% respectively. Multivariate analysis showed the GS12 to be an independent risk factor for the progression of MGUS to MM. Comparing GS12 with GEP70 showed that GS12 is better at predicting progression to MM than GEP70. GEP70-defined high-risk MGUS predicted risk of progression to MM of 18% while GS12-defined high-risk MGUS raised that estimate to 73.9%. Conclusion: A gene score based upon twelve highly differentiating genes showed excellent accuracy in predicting 10-year MM progression from MGUS. Integration of gene signatures into the routine management of MGUS patients may improve the application of risk-adapted approaches to prevent progression to overt MM. We are in the process of collecting more samples and incorporating RNA-seq data to determine whether this model can be adapted to predict MGUS progression on a larger scale with current state-of-the-art technologies.

Synovial calprotectin is a reliable biomarker for periprosthetic joint infections in acute-phase inflammation — a prospective cohort study

PurposeDiagnosing periprosthetic joint infections (PJI) are challenging and may be hampered by the presence of other causes of local inflammation. Conventional synovial and serum markers are not reliable under these circumstances. Synovial calprotectin has been recently shown as a promising biomarker for PJI in total hip (THA) and total knee arthroplasty (TKA). The aim of this study is to investigate if calprotectin is reliable for PJI diagnosis in cases with accompanying inflammation due to recent surgery, dislocation or implant breakage in primary and revision TKA and THA.MethodsThirty-three patients were included in this prospective study between July 2019 and October 2021 (17 patients undergoing surgery < 9 months, 11 dislocations, five implant breakage, respectively). Synovial white blood cell count (WBC), percentage of polymorphonuclear neutrophils (PMC), serum C-reactive protein (CRP) and synovial calprotectin, using a lateral-flow-assay, were analysed. These parameters were tested against a modified European-Bone-and-Joint-Infection-Society (EBJIS) definition with adjusted thresholds to account for the local inflammation. Statistic quality criteria were calculated and compared using a binary classification test.ResultsSeventeen patients were classified as confirmed infections according to the modified EBJIS definition (13 THA and 4 TKA). The calprotectin assay yielded a sensitivity of 0.88 (0.64, 0.99), a specificity of 0.81 (0.54, 0.96), a positive predictive value (PPV) of 0.83 (0.59, 0.96) and a negative predictive value (NPV) of 0.87 (0.60, 0.98).ConclusionsEven in the presence of local inflammation due to other, non-infectious causes, calprotectin is a reliable diagnostic parameter for the detection of a PJI in primary and revision THA and TKA.

Urinary Neutrophil Gelatinase-Associated Lipocalin as a Novel Predictor in Treatment of Active Lupus Nephritis

Abstract Introduction Lupus nephritis (LN) affects almost two-thirds of Systemic lupus erythematosus (SLE) patients. Despite initial aggressive therapy, up to 25% of patients with LN will progress to end stage renal disease (ESRD). Conventional serum markers for LN lack the sensitivity of an ideal biomarker. Urinary Neutrophil gelatinase-associated lipocalin (UNGAL) is an excellent biomarker for early diagnosis of acute kidney injury and predicting renal outcomes. Objectives To measure UNGAL among LN patients correlating its levels with renal disease activity to investigate its predictive performance in response to induction therapy. Patients and Methods 40 SLE patients with biopsy-proven LN, class III, IV, or V were randomly selected. The study was conducted in internal medicine department and Outpatient clinic in Ain Shams University Hospitals on and completed after six months. UNGAL was measured at baseline and after complete induction therapy. Results In LN patients at baseline; mean UNGAL levels of complete response, partial response, and no response groups were 14.48 ±2.99 ng/ml, 34.49 ±4.09, and 62.07 ±14.44 ng/ml respectively. Based on ROC curve, we found better performance of baseline UNGAL to discriminate complete response group from partial and non-response groups to predict response to induction, outperforming conventional biomarkers. The area under the curve was 0.943 (92.31% sensitivity, 88.89% specificity), and the best cut-off level was 26.5 ng/ml. Conclusion UNGAL performed better than conventional biomarkers in predicting response to treatment of active LN.

Laboratory Assessment of Disease Activity in Pediatric Patients with Inflammatory Bowel Disease: What’s New?

Laboratory tests are an integral part of both the diagnostic and follow-up algorithm of patients with inflammatory bowel disease (IBD). Their advantages over other non-invasive methods for assessing disease activity are greater objectivity than clinical activity indices and imaging studies. This review aims to analyze shortly the most common laboratory tests used to assess disease activity in pediatric patients with IBD. In addition to the conventional blood and serum markers that are not specific for gut inflammation, although routinely used, we also reviewed the established fecal markers such as calprotectin, lactoferrin, M2-pyruvate kinase, osteoprotegerin, HMGB1, chitinase 3-like 1, and the promising non-coding microRNA. In conclusion, neither marker is unique to the pediatric IBD. More clinical data are required to assess biomarkers’ full potential for diagnosis, management, and follow-up of pediatric IBD patients.

Value of Urinary Neutrophil Gelatinase-Associated Lipocalin versus Conventional Biomarkers in Predicting Response to Treatment of Active Lupus Nephritis.

Introduction Lupus nephritis (LN) affects almost two-thirds of systemic lupus erythematosus (SLE) patients. Despite initial aggressive therapy, up to 25% of patients with LN will progress to permanent renal damage. Conventional serum markers for LN lack the sensitivity of an ideal biomarker. Urinary neutrophil gelatinase-associated lipocalin (UNGAL) is an excellent biomarker for early diagnosis of acute kidney injury and predicting renal outcomes. Objective To measure UNGAL among LN patients to correlate its levels with renal disease activity and to investigate its predictive performance in response to induction therapy. Patients and Methods. 40 SLE patients with biopsy-proven LN class III, IV, or V were randomly selected. The study was conducted in the internal medicine department and outpatient clinic in Ain Shams University Hospitals and completed after six months. UNGAL was measured at baseline, three-month follow-up, and after complete induction therapy. Results In LN patients at baseline, the mean serum creatinine was 2.57 ± 0.96 mg/dL and the mean UNGAL was 33.50 ± 18.34 ng/dL. Mean UNGAL levels of complete response, partial response, and nonresponse groups were 14.48 ± 2.99 ng/mL, 34.49 ± 4.09 ng/mL, and 62.07 ± 14.44 ng/mL, respectively. Based on the ROC curve, we found a better performance of baseline UNGAL to discriminate the complete response group from partial and nonresponse groups to predict response to induction, outperforming conventional biomarkers. The area under the curve was 0.943, and the best cutoff level was 26.5 ng/mL (92.31% sensitivity and 88.89% specificity). Conclusion UNGAL performed better than conventional biomarkers in predicting response to treatment of active LN.

Extending the provision of cell-free DNA screening for trisomy21.

It is now 12 years since the discovery that fetal cfDNA fragments in maternal plasma can be used to detect Down syndrome. This single marker has a detection rate above 99% and a false-positive rate below 0.2%, a performance greatly exceeding conventional multiple serum and ultrasound marker screening tests. Nevertheless, there has not been widespread replacement of conventional screening by cfDNA. Commercial providers report annual cfDNA testing totaling millions but this covers only a few percent of pregnancies worldwide.

Serum markers alone can define high-risk smouldering multiple myeloma: new insights from the Czech Myeloma Group model.

The overall risk of developing symptomatic multiple myeloma (MM) from the asymtomatic precursor state, smoldering multiple myeloma (SMM), is 10% per year, but varies greatly with the presence of certain risk factors.1 The heterogeneous clinical course of SMM has led the International Myeloma Working Group (IMWG) to propose that patients with an 80% risk of progression at two years after an SMM diagnosis should be regarded as symptomatic MM patients and offered treatment.2 Therefore, the updated diagnostic MM criteria included three biomarker sassociated with a very high risk of progression; BMPC% ≥60, sFLCr ≥100 and/or more than one focal lesion on MRI.2 Within the last decades, several variables have been identified as associated with an increased risk of SMM progression to MM. Most risk-models are composed of a combination of serum markers3, 4 or a combination of serum markers along with bone marrow biopsy5, 6 flow-cytometry,7 imaging8 or molecular markers.9, 10 However, there is still no international consensus on which factors should be used as the standard risk stratification. Furthermore, one study found poor consistency between two widely used risk-models (PETHEMA and Mayo Clinic) when applied to the same patient cohort.11 Also, in the Danish study we found that the two-year progression-risk of having an abnormal FLC ratio ≥100 was only 30% and thus nowhere near the >80% risk as shown in the two studies which formed the basis of the IMWG guidelines.4 There are two important problems with the different SMM risk-models published so far: (i) the models are usually based on single-centre observations, and (ii) not all models consist of easily accessible markers for risk stratification. Together, this increases the risk of bias and to a certain degree compromises the ability of the risk-models to be used in the broader SMM population. It seems reasonable to assume that the lack of reproducibility of SMM high-risk models is also influenced by the biological heterogeneity of precursor diseases. A study analysing the whole genome sequence of pared tumour samples from 10 patients progressing from SMM to MM showed that the complexity of the genomic profile is generally not associated with a shorter time-to-progression (TTP) and that both a static and spontaneous sub-clonal evolution was found among the tested patients.12 How conventional serum and/or bone marrow markers serve as the correct proxy for the underlying genomic profile of the individual SMM patient is unknown. In this issue of the journal, Hájek et al. presents a new risk-model based on the data from the Czech Myeloma Group (CMG) and Heidelberg group. Based on data from the nationwide CMG registry, the authors first identified a high-risk subpopulation of SMM patients with a 78·7% two-year progression risk using immunoparesis, M-protein level ≥2·3 g/l and a FLC ratio >30 risk factors only. The risk-model was subsequently tested externally in the Heidelberg cohort, which arguably represents a specialised MM centre. Interestingly, the authors found a remarkably similar two-year progression risk (81·3%) when the model was tested in the Heidelberg population. Furthermore, Hájek et al. used the same data for a post-hoc comparison with other published risk-models and found that the CMG data did not meet the 80% progression rate threshold, indicating that the CMG model is more robust than previously proposed risk-models. Do we need more risk-models for SMM? With the QuiRedex study we saw that upfront treatment with lenalidomide-dexamethasone of high-risk patients does prolong TTP and overall survival (OS).13 Side effects were fairly modest with few grade 3 events.13 Arguably, if patients have a better OS and side effects are manageable, then the risk of doing more harm than good when starting anti-MM treatment is low. However, if high-risk SMM patients with high-risk features are to be considered as symptomatic MM patients, standard treatment should arguably involve three-drug induction treatment followed by ASCT for younger as well as fit elderly patients. Recently, daratumumab monotherapy for high-risk SMM patients was evaluated in the phase 2 Centaurus trial.14 The study did not meet the co-primary endpoint of CR >15%, thus indicating that a more aggressive multi-drug regimen is probably needed for disease eradication. Several clinical trials using different risk-models are underway to evaluate intensive treatment approaches in SMM. A real strength with the new CMG model is the external validation and use of accessible serum risk factors which makes this model practical for inclusion in future clinical trials or for the evaluation of SMM patients in everyday clinical practice.

Using the polymeric circulating tumor cell chip to capture circulating tumor cells in blood samples of patients with colorectal cancer.

The current study clarified the accuracy of a circulating tumor cell (CTC) detection system to diagnose colorectal cancer using blood samples. The system uses the ‘polymeric CTC-chip,’ (CTC-chip), which is a microfluidic device that is used for CTC isolation. CTCs are considered sensitive diagnostic biomarkers. However, their concentration in the peripheral blood is low and requires highly sensitive and specific capturing techniques. The capture efficiency of the polymeric CTC-chip was first assessed using cell suspensions of the colorectal cancer cell line HCT-116, which was reported as 90.9% in a phosphate-buffered saline suspension and 65.0% in the blood. The CTC-chip was then used to detect CTCs in blood samples obtained from 13 patients with stage II–IV colorectal cancer. On average, the CTCs/ml was lower in patients with stages II and III colorectal cancer (3.3±2.3) than in those with stage IV (7.0±6.2). In patients with stages II–IV, 92% had ≥1 CTC per ml, which was significantly higher than the positive rate (15%) detected using the carbohydrate antigen 19-9 test (CA19-9). Furthermore, CTCs were detected in all patients with stage II and III colorectal cancer, including a number of patients with negative results for the carcinoembryonic antigen (CEA) and CA19-9 tests. With the polymeric CTC-chip detection system, CTCs can be effective cancer markers, particularly for patients with stage II and III colorectal cancer who often exhibit negative conventional serum marker test results. The CTC-chip system may also facilitate the detection of cancer progression based on CTC concentration.

Fungal Infections of the Central Nervous System.

Fungal central nervous system infections present in a myriad of clinical manifestation and remain a significant diagnostic challenge. Presenting symptoms, cerebrospinal fluid analysis, conventional fungal serum markers and imaging correlates of disease are seldom sensitive or specific; fungal culture growth or histopathologic analysis are often required for definitive diagnoses but limited by feasibility. Novel diagnostic tools (such as the cryptococcal antigen and next-generation sequencing) have increased diagnostic potency when available but require further studies to define their utility.Invasive fungal infections are an emerging health threat in the setting of increased immunomodulatory treatments, advancements in transplant medicine and increased world-wide travel. Therefore, strong clinical suspicion from epidemiologic clues, clinical progression and presence of CNS dissemination risk factors must be exercised to pursue broad diagnostic workups and rapidly initiate medical and surgical management. This article will describe the epidemiology, clinical presentations, diagnostic approach and therapeutic interventions for fungal infections in the central nervous system.

Lipoprotein-associated phospholipase A2 in relation to lipid markers: A pan-India correlation study

Background: Conventional serum markers are routinely recommended to assess heart health. However, many new biochemical markers like lipoprotein-associated phospholipase A2 (Lp-PLA2) has been studied as an efficient cardiac risk marker. Our report is an attempt to present correlation between traditional lipid markers and Lp-PLA2 in a pan-India cohort.&#x0D; Materials and Methods: The study cohort included a total of 10347 pan-Indians including 5945 males and 4402 females respectively. The serum lipid parameters estimated include triglycerides, LDL- and HDL-cholesterol as well as total cholesterol by the technology of photometry.&#x0D; Results: The frequency of high Lp-PLA2 detected in our study was 6.4% and the frequency between males at 9.3% and females at 2.5% was detected to be significant at p&lt;0.0001. Linear regression analysis detected significant positive association of high Lp-PLA2 with LDL-cholesterol and total cholesterol at p&lt;0.05, while the trend with HDL-cholesterol was also significant. However, Lp-PLA2 as an independent marker was not detected to be significant in relation to the lipid indices studied.&#x0D; Conclusion: Though Lp-PLA2 has been associated with risk factor for acute coronary crisis, its clinical value remains controversial. Our report though detected significant association of Lp-PLA2 with various lipid parameters, independent value remains questionable.

82TiP - SCAN-B-rec: Infrastructure, technology and clinical research platform to profile and monitor metastatic breast cancer

Background: Great advances have been made in the molecular characterisation of advanced breast cancer (ABC). Still, clinical practice has not changed in terms of assessment of response to therapy. Radiologic imaging and clinical examination help determine disease burden and treatment efficacy. However, imaging heavily relies on the expertise of the radiologist and oncologist and has high intra- and inter-reader variability. Circulating tumor cells (CTCs) enumeration with CellSearchâ has confirmed independent prognostic value compared to conventional radiology and serum markers such as CA15-3. Similarly, quantification of circulating tumor DNA (ctDNA) has also prognostic value in ABC. ctDNA analysis further provides critical information on the mutational status of specific genes which can predict sensitivity to specific targeted therapeutics. Trial design: The SCAN-B-rec (NCT03758976) is a prospective multi-center observational study that aims to explore the value provided by ctDNA analysis to determine treatment efficacy when compared to CTCs and conventional radiological imaging in a “real-world” setting. A total of 350 patients with pathologically-confirmed ABC will be recruited. Plasma ctDNA will be extracted and analysed before treatment start (T0) and after three months of therapy (T1). The relative change in mutant allele frequency (MAF) between these two time-points will be calculated. Response to treatment will be defined using pre-specified cut-offs. A comparison with response to treatment according to conventional radiological imaging (RECIST criteria) and CTC enumeration at the same time points will be performed. The primary objective is to establish the prognostic value of ctDNA to predict overall survival. The study will also explore the added value provided by ctDNA mutational analysis to determine biomarkers of sensitivity and resistance to targeted therapeutics approved to treat ABC or other solid tumors that could be used “off-label”. We expect that ctDNA quantification will give a more accurate read-out of treatment response, providing better prognostic information in ABC as well as information on drug sensitivity/resistance in a large proportion of ABC patients. Clinical trial identification: SCAN-B-rec (NCT03758976). Legal entity responsible for the study: Lund University Hospital (Region Skane). Funding: BioCARE, Cancer Fonden, ALF (Region Skåne) and Svenska Läkaresällskapet. Disclosure: A. Bosch: Advisory board meetings: Pfizer, Novartis in Sweden. These have been non-remunarated. L. Saal: CEO: AGA Diagnostics. All other authors have declared no conflicts of interest.

Whole-body Imaging of Cell Death Provides a Systemic, Minimally Invasive, Dynamic, and Near-real Time Indicator for Chemotherapeutic Drug Toxicity.

Response to toxicity in chemotherapies varies considerably from tissue to tissue and from patient to patient. An ability to monitor the tissue damage done by chemotherapy may have a profound impact on treatment and prognosis allowing for a proactive management in understanding and mitigating such events. For the first time, we investigated the feasibility of using whole-body imaging to map chemotherapeutic drug-induced toxicity on an individual basis. In a preclinical proof-of-concept, rats were treated with a single clinical dose of cyclophosphamide, methotrexate, or cisplatin. In vivo whole-body imaging data were acquired using 99mTc-duramycin, which identifies dead and dying cells as an unambiguous marker for tissue injury in susceptible organs. Imaging results were cross-validated using quantitative ex vivo measurements and histopathology and compared with standard blood and serum panels for toxicology. The in vivo whole-body imaging data detected widespread changes, where spatially heterogeneous toxic effects were identified across different tissues, within substructures of organs, as well as among different individuals. The signal changes were consistent with established toxicity profiles of these chemotherapeutic drugs. Apart from generating a map of susceptible tissues, this in vivo imaging approach was more sensitive compared with conventional blood and serum markers used in toxicology. Also, repeated imaging during the acute period after drug treatment captured different kinetics of tissue injury among susceptible organs in males and females. This novel and highly translational imaging approach shows promise in optimizing therapeutic decisions by detecting and managing drug toxicity on a personalized basis.Toxicity to normal tissues is a significant limitation in chemotherapies. This work demonstrated an in vivo imaging-based approach for characterizing toxicity-induced tissue injury in a systemic, dynamic, and near-real time fashion. This novel approach shows promise in optimizing therapeutic decisions by monitoring drug toxicity on a personalized basis.

Relationship between serum vascular cell adhesion molecule-1 and endothelin-1 levels with organ involvement and disease activity in systemic lupus erythematosus patients.

Background Endothelial dysfunction plays an important role in pathogenesis of systemic lupus erythematosus (SLE). Considering the importance of serum soluble vascular cell adhesion molecule-1 as the most abundant of the circulating adhesion molecules increased as a result of endothelial dysfunction and the role of endothelin-1 in pathophysiology of SLE, this study aimed to evaluate serum soluble vascular cell adhesion molecule-1 and endothelin-1 levels in SLE patients compared to healthy subjects. Methods In this cross-sectional study, 60 SLE patients according to the Systemic Lupus International Collaborating Clinics classification criteria for SLE and 40 age and sex-matched healthy controls were included. In patients, clinical examination was performed and SLE disease activity index was assessed. Serum endothelin-1 and soluble vascular cell adhesion molecule-1 levels were measured using ELISA kits. Results The mean ± standard deviation age of patients and controls was 31.91 ± 7.66 and 33.20 ± 10.08 years, respectively. Compared to healthy controls, serum soluble vascular cell adhesion molecule-1 (1023.8 ± 352.96 vs. 866.06 ± 109.91) and endothelin-1 (77.83 ± 16.27 vs. 54.45 ± 12.01) was significantly higher in SLE patients ( P = 0.003 and P < 0.001, respectively). The most common organs involved in patients were skin, joint and kidney. There were no significant differences in serum soluble vascular cell adhesion molecule-1 and endothelin-1 levels according to organ involvement, activity of disease and the conventional serum markers of disease activity ( P > 0.05). There was no significant correlation between disease activity, organ involvement and negative or positivity of autoantibodies as well as serum complement with endothelin-1 and vascular cell adhesion molecule-1 levels ( P > 0.05). Conclusions Although our study revealed higher serum soluble vascular cell adhesion molecule-1 and endothelin-1 levels in SLE patients compared to healthy controls, there were no significant correlations between their serum levels with organ involvement and disease activity.

Non-invasive prediction of NAFLD severity: a comprehensive, independent validation of previously postulated serum microRNA biomarkers

Liver biopsy is currently the only reliable method to establish nonalcoholic fatty liver disease (NAFLD) severity. However, this technique is invasive and occasionally associated with severe complications. Thus, non-invasive diagnostic markers for NAFLD are needed. Former studies have postulated 18 different serum microRNA biomarkers with altered levels in NAFLD patients. In the present study, we have re-examined the predictive value of these serum microRNAs and found that 9 of them (miR-34a, -192, -27b, -122, -22, -21, -197, -30c and -16) associated to NAFLD severity in our independent cohort. Moreover, miR-192, -27b, -22, -197 and -30c appeared specific for NAFLD, when compared with patients with drug-induced liver injury. Preliminary serum RNAseq analysis allowed identifying novel potential miRNA biomarkers for nonalcoholic steatohepatitis (NASH). The classification performance of validated miRNAs (and their ratios) for NASH was better than that reached by AST, whereas for advanced fibrosis prediction miRNAs did not perform better than the FIB-4 algorithm. Cross-validated models combining both clinical and miRNA variables showed enhanced predictivity. In conclusion, the circulating microRNAs validated demonstrate a better diagnostic potential than conventional serum markers to identify NASH patients and could complement and improve current fibrosis prediction algorithms. The research in this field is still open.

Serum p53 antibody as a potential tumor marker in extrahepatic cholangiocarcinoma.

Only a few studies have evaluated the clinicopathological significance of the p53 protein expression and s-p53-Abs level in patients with cholangiocarcinoma. We therefore analyzed the clinicopathological and prognostic significance of s-p53-Abs in patients with extrahepatic cholangiocarcinoma. We prospectively evaluated s-p53-Abs levels before and after surgery in 61 patients with extrahepatic cholangiocarcinoma to determine the relationship between clinicopathological factors and the prognostic significance of s-p53-Abs. Among a total of 61 primary extrahepatic cholangiocarcinoma cases, 23% were positive for s-p53-Abs. Combination of s-p53-Abs with the conventional serum markers carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) significantly increased the rate of positive extrahepatic cholangiocarcinoma cases (57% for CEA and/or CA19-9 vs. 75% for CEA and/or CA19-9 and/or s-p53-Abs, P=0.035). There were no significant differences in clinicopathological factors between the p53-seropositive and p53-seronegative patients. An immunohistochemical analysis showed the presence of significant associations between the intensity (P=0.003) and extent (P=0.001) of p53 immunoreactivity and p53-seropositivitly. Although s-p53-Abs was not a significant prognostic factor for the survival in either univariate or multivariate analyses, p53 immunoreactivity was independently associated with a poor survival. Among patients positive for s-p53-Abs before surgery, the s-p53-Abs levels were reduced after surgery in most. These findings suggested that s-p53-Abs might be associated with p53 immunoreactivity. In addition, s-p53-Abs may be useful for a diagnosis, but was not useful for predicting tumor recurrence or the survival. This study was registered as UMIN000014530.