Blood Plasma Biomarkers Research Articles

New diagnostic markers in basal cell carcinoma.

Background:The clinical significance of plasma lipoprotein levels in the diagnosis and prognosis of certain diseases is known fact. Some studies have reported different and contradictory levels of blood lipoproteins in cancer patients. Therefore, we decided to compare lipid profiles in patients with basal cell carcinoma (BCC) and healthy individuals.Materials and Methods:In this case-control study, lipid biomarkers in blood plasma of 64 patients with BCC compared with biomarkers in 64 healthy subjects. According to including criteria's both group selected. 5 cc blood samples taken after 8–12 h of fasting. Ultimately, the diagnosis of biopsy samples confirmed by the dermatopathologist. The information data entered into the PASS II software and then characterized by a descriptive and analytical statistics of the lipid profile. The role of the related factors was assessed by two sample t-test power analysis.Results:In this research, mean age of patient group and the healthy one was 67.13 ± 11.33 and 64.31 ± 6.98, respectively. The average amount of triglyceride, cholesterol, high density lipoprotein (HDL) and low density lipoprotein in the BCC patients were 147.97 ± 87.11 mg/dl, 188.25 ± 38.90 mg/dl, 61.98 ± 18.61 mg/dl and 95.98 ± 31.69 mg/dl, respectively, whereas these amounts in the control group were 137.34 ± 61.41 mg/dl, 173.22 ± 38.79 mg/dl, 42.34 ± 7.83 mg/dl and 106.44 ± 35.17 mg/dl, respectively. There were statistically significant differences between cholesterol and HDL in patients with BCC and healthy controls, respectively (P = 0.030) (P < 0.001).Conclusion:It seems that changes in lipid profile can help as a diagnostic marker for detecting cancer like BCC. Although this change could be different in lipid markers and also among different types of cancer.

The Role of Inflammation after Surgery for Elders (RISE) study: Examination of [11C]PBR28 binding and exploration of its link to post-operative delirium

Major surgery is associated with a systemic inflammatory cascade that is thought, in some cases, to contribute to transient and/or sustained cognitive decline, possibly through neuroinflammatory mechanisms. However, the relationship between surgery, peripheral and central nervous system inflammation, and post-operative cognitive outcomes remains unclear in humans, primarily owing to limitations of in vivo biomarkers of neuroinflammation which vary in sensitivity, specificity, validity, and reliability. In the present study, [11C]PBR28 positron emission tomography, cerebrospinal fluid (CSF), and blood plasma biomarkers of inflammation were assessed pre-operatively and 1-month post-operatively in a cohort of patients (N = 36; 30 females; ≥70 years old) undergoing major orthopedic surgery under spinal anesthesia. Delirium incidence and severity were evaluated daily during hospitalization. Whole-brain voxel-wise and regions-of-interest analyses were performed to determine the magnitude and spatial extent of changes in [11C]PBR28 uptake following surgery. Results demonstrated that, compared with pre-operative baseline, [11C]PBR28 binding in the brain was globally downregulated at 1 month following major orthopedic surgery, possibly suggesting downregulation of the immune system of the brain. No significant relationship was identified between post-operative delirium and [11C]PBR28 binding, possibly due to a small number (n = 6) of delirium cases in the sample. Additionally, no significant relationships were identified between [11C]PBR28 binding and CSF/plasma biomarkers of inflammation. Collectively, these results contribute to the literature by demonstrating in a sizeable sample the effect of major surgery on neuroimmune activation and preliminary evidence identifying no apparent associations between [11C]PBR28 binding and fluid inflammatory markers or post-operative delirium.

Muscle-tendon weakness contributes to chronic fatigue syndrome in Gaucher\u2019s disease

BackgroundChronic fatigue (CFg) is a prevalent symptom in Gaucher disease (GD) at diagnosis (79%) and remains in a quarter of patients after years of therapy. Bone abnormalities are present in over 70% and peripheral neuropathy in about 11% of the patients, which contributes to the disabling and debilitating complications. Our hypothesis is that other factors such as muscle-tendinous weakness could have influence in the development of CFg.MethodsWe have evaluated the fiber structure and elasticity of muscle-tendinous unit by strain-elastography (S-ELA) and analyzed their influence in the CFg. S-ELA study was performed in Achilles tendon in 25 type 1 and two type 3 GD patients, all of them with fatigue and were on enzymatic replacement therapy for mean 13 years; simultaneously, bone marrow burden by MRI and calcaneus ultrasound densitometry were evaluated. Blood cell counts, plasma biomarkers, GBA1 genotyping, and SF36 quality of life scale (QoL) were also performed. Statistical analysis: descriptive and comparative test.ResultsAll patients showed a normal Achilles tendinous structure. Abnormal stiff grade 2–3 was found in 17/27 (62.9%); in 11/27 (40.7%) of patients, the alteration was bilateral. There were no correlations between the S-ELA results to other variables; nevertheless, a significant correlation between the degree of tendon hardness and the low score on the QoL scales (p = 0.0035) was found. The S-ELA is a sensitive painless, fast, and low cost method to detect muscle-tendinous subclinical dysfunction that could contribute to CFg in GD. The identification of subclinical tendon alteration would be a sign of alarm, focused on the risk of development of bone complications.ConclusionIntratendinous alteration in strain-elastography is an independent variable in GD patients with persistent fatigue.

Assessing the seasonal and intrinsic variability of neurotoxic and cyto-genotoxic biomarkers in blood of free-living Eleonoras’ falcons

In the present study we investigated seasonal and intrinsic variability of neurotoxic and cyto-genotoxic biomarkers in blood plasma and erythrocytes of free-living Eleonoras’ falcons, captured during the pre-breeding (May of 2017 and 2018) and breeding period (September of 2017) on the Antikythira Island (Greece). Specifically, blood samples of captured birds were prepared for the determination of cholinesterase (ChEs, i.e. acetylcholinesterase/AChE and butyrylcholinesterase/ BChE) activity, as well as the formation of nuclear (i.e. the formation of micronuclei into the cells/MN, binucleated cells/BN and others), and cellular/cytoplasmic (i.e. echinocytes/EC, acanthocytes/AC and notched cells/NC) abnormalities in blood plasma and erythrocytes, respectively. Our results indicated that birds sampled in late May had higher ChE and BChE activity levels, as well as higher frequency of total nuclear abnormalities. The latter were also higher in second calendar year (2cy) birds. Cellular/cytoplasmic abnormalities were less frequent in falcons having better body condition, sampled in late May, as well as in light-morph falcons. The observed ChEs activities, as well as nuclear and cellular/cytoplasmic abnormalities revealed that Eleonora’s falcons are likely to be exposed to chemical agents with neurotoxic and cyto-genotoxic potential year round, while different aspects of their biology and ecology, such as their reproductive and nutritional status, could mediate their levels. Although we encourage more sampling campaigns to verify the identified seasonal and intrinsic sources of variation in biomarkers tested, the current study enriches the existing knowledge about their usefulness in the environmental monitoring and risk assessment of migratory birds, like Eleonoras’ falcon.

Paper-Based Lateral Flow Immunoassay for Detection of Biomarkers in Human Fluids

Traumatic brain injuries (TBI) patients suffer from neurological deficits and increased risk of neurological disease, so there is a need for accurate, high throughput, and rapid diagnosis. Currently, TBI severity is detected and assessed using neurological examination and CT imaging, which require expensive hospital and doctors office visits. Protein biomarkers have emerged as an objective means to diagnose TBI; however, proteins are typically detected using time consuming and expensive methods such as ELISA and Western blot. In this work, a paper-based lateral flow assay (PLFA) has been developed for a rapid, low-cost means for detecting TBI biomarkers in blood plasma. PLFAs have received increasing attention due to their low cost and ease-of-use. However, these devices suffer from poor sensitivity and interference from the blood plasma matrix. In this study, surface-enhanced Raman scattering (SERS) was applied into a PLFA as the signal transducer to improve sensitivity and overcome biological interference. These advantages are attributed to the optical excitation in the near-infrared biological transparency window and SERS nanostructure design. The PLFA is designed using a plasmonic nanoarray chip and a blood plasma separation unit for highly sensitive TBI biomarker detection.

PB1975 CHRONIC FATIGUE, ANAEMIA AND MUSCULOSKELETAL AFFECTATION IN GAUCHER DISEASE

Background:Chronic fatigue is the most prevalent symptom in type 1 Gaucher disease(GD1) (79%) and persists in spite of prolonged therapy which usually achieves the general goals (24.5%). It is mainly attributed to hypermetabolism, myopathy or increase in plasma cytokine concentration. There are several factors such as anaemia, pain, depression, anxiety, sleep disturbances, emotional distress, activity level or medication side effects that could be involved. Bone abnormalities are present in over 70% of the patients and peripheral neuropathy in about 11% of them, which contributes to the disabling and debilitating complications. Our hypothesis is that other factors including muscle‐tendinous weakness could have influence in the development of fatigue.Aims:The aim of the study is to evaluate for first time the study the fiber structure in muscle‐tendinous unit and compromised elasticity usefulness of real‐time tissue Strain‐Elastography in the assessment of the Achilles tendon fibres in GD patients with or without bone marrow infiltration by MRI and their influence in the chronic fatigue.Methods:We have study the fibrillar structure (rigidity) of muscletendinous tissues in Achilles tendon applying a noninvasive technique based on Strain‐Elastography (Hitachi system EUB‐8500, L54 M transducer, frequency 6‐13 MHz) and combined this exam with bone marrow burden study using Spanish‐MRI score and calcaneus ultrasound densitometry for bone mineral density evaluation. We have selected 21 GD1 and 2 type 3 patients (mean age: 46.9 y, 18‐65, 12 females) with chronic fatigue, on enzymatic replacement therapy (mean time 12.0 years, 2‐25). Non history of diabetes, endocrine diseases or concomitant steroid therapy were present. Control group: 20 healthy subjects matched by gender and age. Correlation study with blood cell counts, plasma biomarkers, genotype, concomitant diseases and SF36 quality of life scale (QoL) were performed. A longitudinal and axial scans was performed in each Achilles tendon in proximal, middle and distal third.Results:Mean Hb (g/dL) in females: 13.3 (12.2‐14.1), in males: 14.8 (13.0‐17.3), advanced bone disease 10 patients (43.5%), S‐MRI (5.1 points; 0‐18), Osteopenia 8 patients (34.7%), T‐score (mean ‐0.76, ‐2.68‐1.51), Z‐score (mean ‐0.53, ‐2.43‐1.45), genotype N370S/L444P (47.8%), N370S/other (34.7%), L444P/other (17.4%), Ferritin: 310.7 (30‐2000), Chitotriosidase (1,813 nmol/mL/h 50‐6,575). CCL18/PARC (256.3 ng/mL 55‐553), Lyso‐Gb1 (32.9 ng/mL 0‐144). All patientes have a normal tendinous structure (mode B). Intratendinuous alteration in Strain‐elastography (S‐ELA) grade 2‐3 was found in 14 /23 (61%) patients and bilateral in 9/23 (21.7%). There are not correlation between S‐ELA grade to other variables. QoL evaluation showed a low score in physical and emotional roles and general health and high in pain. There are a significant correlation between the degree of tendon hardness and the low score on the QoL scales, (p = 0.0035).Summary/Conclusion:In Achilles tendon Strain‐elastography is a sensitive method to detect subclinical disfunction of muscle‐tendinous that could contribute to fatigue symptom in GD. Intratendinuous alteration is one independent variable in GD patients. The cause of this disfunction probably has a mechanical origen and can be a risk factor for the development of skeletal complications. A project based on physical activity has been developed to Increase the functional strength of the lower limbs achieve greater stability, power and improvement in physical activities, as well as the reduction of fatigue.

Identification of potential endometriosis biomarkers in peritoneal fluid and blood plasma via shotgun lipidomics

Endometriosis is a recurrent and benign gynecological disorder, defined by the ectopic presence of endometrium. About 10% of reproductive-aged women suffer from endometriosis. There are no non-invasive or minimally invasive tests available in clinical practice to accurately diagnose endometriosis today.Here, we present our efforts to determine the diagnostic accuracy of biomarkers in peritoneal fluid and blood plasma using flow injection analysis with electrospray ionization tandem mass spectrometry (ESI-MS/MS) in 70 women with endometriosis and 20 women from a control group. The presence of endometriosis was confirmed by surgical findings and post-operative pathological examination. A qualitative and quantitative evaluation of the lipids in peritoneal fluids and blood plasma was carried out using electrospray ionization mass spectrometry (ESI-MS). The analysis revealed more than 140 molecular species of lipids, most of which pertained to five classes: phosphatidylcholines, phosphatidylethanolamines, sphingomyelins, di- and triglycerides. The data were analyzed using a statistical multifactorial method (i.e., PLS-DA). It was found that 9 potential biomarkers of endometriosis (LPC 16:0, PE O-20:0, PE O 34:1, PC 36:2, PC 36:4, PC 36:5, PC 38:4, PC 38:6 and SM 34:1) are common in blood plasma and peritoneal fluid, supporting connection with the pathological process. The sensitivity of the method developed for plasma was 93% with a specificity of 95%; for peritoneal fluid, the sensitivity was 90% and the specificity 95%. Accordingly, plasma is the most suitable biological fluid for clinical diagnostics of endometriosis. Further validation of these lipids as serologic biomarkers may enhance non-invasive diagnostic tools for patients with suspected endometriosis and reduce the frequency of diagnostic laparoscopy.

Hydration status and health

Water is an important component of human body and plays a variety of important physiological roles. The intake and discharge of human water is in a dynamic equilibrium. Insufficient water intake will affect the hydration status of human body, which in turn affects cognition and health. Therefore, maintaining proper hydration status is of great importance for maintaining and promoting the health of the body. There were many indicators to evaluate the hydration status of body, including urine and blood plasma biomarkers. It is of great significance to evaluate the hydration status of body, keep the body in a suitable hydration status by replenishing water in time. The lack of drinking water in different populations in China is common, but it has not yet received enough attention and needs to carry out corresponding health education.

A Nanoplasmonic Biosensor for Ultrasensitive Detection of Alzheimer's Disease Biomarker Using a Chaotropic Agent.

Blood-based diagnosis (hemodiagnosis) of Alzheimer's disease (AD) is emerging as a promising alternative to cerebrospinal-fluid-based methods because blood contains various kinds of AD biomarkers, including amyloid beta 1-40, 1-42, and τ (tau) protein. However, with current technology, the accuracy of the blood-plasma-based methods is relatively low compared to the traditional methods because the concentration of AD biomarkers in blood plasma is incredibly low, and diverse interference is present in blood plasma, which hinders precise detection. Here, we suggest a nanoplasmonic biosensor using gold nanorods with a chaotropic agent for precise ultrasensitive detecting of Alzheimer's disease biomarkers in human plasma. This nanoplasmonic biosensor is based on the localized surface plasmon resonance (LSPR), which is extremely sensitive to the point where it can respond to an insignificant change of the refractive index around the gold nanoparticles. Also, using guanidine hydrochloride as a chaotropic agent, we can overcome the obstacles of blood-based AD diagnostics. In more detail, this agent interrupts the network between water molecules and weakens the hydrophobic interactions between proteins, remarkably improving detection capabilities to target τ protein. By reducing the overlapping ranges between protein levels in an age-matched control and AD patients' plasma, this system can accurately diagnose AD patients. This platform also can analyze disease from mild cognitive impairment using standardized blood biomarker tau protein, which is related to Alzheimer's disease. As a result, our platform can be applied to clinical trials, and thus it has excellent potential in the medical field.

Abstract 63: Plasma Biomarkers of Cavernous Angioma With Symptomatic Hemorrhage (CASH)

Introduction: Cerebral cavernous angiomas with a documented symptomatic hemorrhage (CASH) are associated with an exceptionally high risk of recurrent hemorrhage and serious morbidity. It is unclear if peripheral blood plasma biomarkers implicated in disease biology can differentiate CASH, and whether the same or different biomarkers can distinguish cases who would rebleed. Methods: Eighteen plasma molecules with postulated mechanistic roles in this disease were quantified in 114 patients and 12 healthy subjects. The best biomarker combination differentiating CASH in the prior year (+/- 30 days) was derived by minimizing the Akaike Information Criterion (AIC), and was validated using statistical and machine-learning simulations. This was compared to the biomarker predicting bleeding in the subsequent year (+/- 30 days), and both explanatory and predictive biomarkers were followed longitudinally in a subset of cases. Results: Nine plasma molecules had significantly different levels expressed in CASH patients (p&lt;0·05, FDR corrected). The best biomarker (AIC=75·9) included a weighted combination of 4 of these (sCD-14, VEGF, CRP, and IL-10), and distinguished CASH patients with 76% sensitivity and 80% specificity (p=0·0003). A weighed combination of two of these compounds (sCD14 and VEGF) and two others (IL-1β and sROBO-4) predicted which patients would bleed in the subsequent year with 83% sensitivity and 93% specificity (p=0·001). The CASH biomarker increased in recovering patients (p=0·01), while the predictive biomarker decreased after a bleed (p=0·01). No significant change over time was observed in stable subjects in either biomarker. Conclusions: Our results suggest shared and unique biomarkers of recent symptomatic hemorrhage and of future bleeding in cavernous angiomas. This may be applied for risk stratification in clinical trials, and potentially developed as a tool in clinical practice.

Elaboration of a method for the determination of low-molecular-weight mustard gas biomarkers in biological samples

The relevance of the development and the improvement of methods for the determination of metabolites (biomarkers) of toxic substances in biological fluids (blood, urine, etc.) is associated with the need to establish facts of humans and animals exposure to toxic chemicals. The need for such analysis arises within the framework of monitoring of the compliance with the provisions of the Chemical Weapons Convention (CWC), when conducting investigations into the alleged use of chemical agents, as well as within the framework of the official proficiency tests carried out by the Organization for the Prohibition of Chemical Weapons (OPCW). Nowadays there exists the problem of the determination of low-molecular-weight mustard gas biomarkers in biological samples with the use of gas chromatography methods with mass-selective detection. The low-molecular-weight biomarkers of sulfur mustard (SM) are thiodiglycol, oxides and sulfoxides. The identification and quantitative assessment of SM markers in blood and urine is carried out according to the methods, based on the displacement of thiodiglycol and its derivatives from protein conjugates with titanium trichloride, solid-phase extraction, concentration in ethyl acetate solution, derivatization with pentafluorobenzoyl chloride, heptafluorobutyrylimidazole, heptafluorobutyric acid anhydride or chloride, subsequent stripping of the derivative into the appropriate solvent and GH-MS analysis (chemical ionization technique with methane as a reagent gas with the registration of negative ions). After the sample preparation, the limits of detection of the minimum values ​​of ultra-low concentrations of the analyzed biomarkers of SM in urine and blood plasma were determined. After the chromatographic analysis, the corresponding graphs of indicators have been constructed, based on the concentrations of biomarkers in urine and blood plasma. Later on they have been used for the elaboration of the methods for the determination of SM biomarkers in human urine and blood.

Raman spectroscopic techniques to detect ovarian cancer biomarkers in blood plasma

Robust diagnosis of ovarian cancer is crucial to improve patient outcomes. The lack of a single and accurate diagnostic approach necessitates the advent of novel methods in the field. In the present study, two spectroscopic techniques, Raman and surface-enhanced Raman spectroscopy (SERS) using silver nanoparticles, have been employed to identify signatures linked to cancer in blood. Blood plasma samples were collected from 27 patients with ovarian cancer and 28 with benign gynecological conditions, the majority of which had a prolapse. Early ovarian cancer cases were also included in the cohort (n = 17). The derived information was processed to account for differences between cancerous and healthy individuals and a support vector machine (SVM) algorithm was applied for classification. A subgroup analysis using CA-125 levels was also conducted to rule out that the observed segregation was due to CA-125 differences between patients and controls. Both techniques provided satisfactory diagnostic accuracy for the detection of ovarian cancer, with spontaneous Raman achieving 94% sensitivity and 96% specificity and SERS 87% sensitivity and 89% specificity. For early ovarian cancer, Raman achieved sensitivity and specificity of 93% and 97%, respectively, while SERS had 80% sensitivity and 94% specificity. Five spectral biomarkers were detected by both techniques and could be utilised as a panel of markers indicating carcinogenesis. CA-125 levels did not seem to undermine the high classification accuracies. This minimally invasive test may provide an alternative diagnostic and screening tool for ovarian cancer that is superior to other established blood-based biomarkers.

Acute Exercise-Induced Circulating Haematopoietic Stem and Progenitor Cells in Cardiac Patients — A Case Series

Exercise-induced circulating haematopoietic stem and progenitor cell (HPC) number has been discussed in the context of regeneration in heart disease patients. The aim of this pilot study was to compare the effect of different exercise protocols usually applied in cardiac rehabilitation on the number of acute, exercise-induced HPCs, related to potential mediators, e.g. biomarkers of sympathetic and oxidative stress, and inflammation. This is a case series comprising seven patients suffering from coronary heart disease (CHD) undertaken at the Center for Ambulant Cardiac Rehabilitation. Patients (n=6) performed two exercise modes (constant-load, CLE; high-intensity interval, HIIE) in randomised order. Venous blood was drawn before and immediately after each test to assess CD34+/CD45+ HPC number by flow cytometry and biomarkers in blood plasma. The primary outcome was the change in HPC number, the secondary outcomes were changes in sympathetic/oxidative stress and markers of inflammation. Both exercise modes resulted in a non-significant increase in HPC number after exercise, even when the results of both tests were combined. Overall, free norepinephrine increased significantly and was positively related to exercise-induced HPC number (r=0.70, p<0.05). Markers of sympathetic activation (fNE), oxidative stress (myeloperoxidase) and inflammation (interleukin-6) significantly increased after CLE and HIIE with no difference between tests. Interestingly, acute CLE and HIIE did not stimulate significant HPC mobilisation in CHD, although both exercise modes elevated circulating concentrations of sympathetic activation. Haematopoietic stem and progenitor cell mobilisation could be blunted due to disease-related bone-marrow exhaustion.

Raman spectral analysis for rapid screening of dengue infection

Infection with the dengue virus is currently clinically detected according to different biomarkers in human blood plasma, commonly measured by enzyme linked immunosorbent assays, including non-structural proteins (Ns1), immunoglobulin M (IgM) and immunoglobulin G (IgG). However, there is little or no mutual correlation between the biomarkers, as demonstrated in this study by a comparison of their levels in samples from 17 patients. As an alternative, the label free, rapid screening technique, Raman spectroscopy has been used for the characterisation/diagnosis of healthy and dengue infected human blood plasma samples. In dengue positive samples, changes in specific Raman spectral bands associated with lipidic and amino acid/protein content are observed and assigned based on literature and these features can be considered as markers associated with dengue development. Based on the spectroscopic analysis of the current, albeit limited, cohort of samples, Principal Components Analysis (PCA) coupled Factorial Discriminant Analysis, yielded values of 97.95% sensitivity and 95.40% specificity for identification of dengue infection. Furthermore, in a comparison of the normal samples to the patient samples which scored low for only one of the biomarker tests, but high or medium for either or both of the other two, PCA-FDA demonstrated a sensitivity of 97.38% and specificity of 86.18%, thus providing an unambiguous screening technology.

The effects of a 12-week worksite physical activity intervention on anthropometric indices, blood pressure indices, and plasma biomarkers of cardiovascular disease risk among university employees

BackgroundTo determine the effectiveness of a low-cost 12-week worksite physical activity intervention targeting a goal of 10,000 steps per day on reducing anthropometric indices, blood pressure indices, and plasma biomarkers of cardiovascular disease (CVD) risk among the employees of a major university.MethodsFifty university employees (n = 43 female, n = 7 male; mean age = 48 ± 10 years) participated in the 12-week physical activity intervention (60 min, 3 day/week). Each session included both aerobic (cardiorespiratory endurance) and muscle-strengthening (resistance) physical activity using existing university facilities and equipment. Anthropometric indices, blood pressure indices, and plasma biomarkers of CVD risk assessed included those for obesity (body mass index), hypertension (systolic blood pressure, SBP; diastolic blood pressure, DBP), dyslipidemia (high-density lipoprotein, HDL; low-density lipoprotein, LDL; total serum cholesterol), and prediabetes (impaired fasting glucose, IFG). Steps per day were assessed using a wrist-worn activity monitor. Participants were given the goal of 10,000 steps per day and categorized as either compliers (≥ 10,000 steps per day on average) or non-compliers (< 10,000 steps per day on average) based on their ability to achieve this goal.ResultsOverall, 34% of participants at baseline were already at an elevated risk of CVD due to age. On average, 28% of participants adhered to the goal of 10,000 steps per day. After 12-weeks, participants in both groups (compliers and non-compliers) had lower BMI scores (p < 0.001), lower HDL scores (p < 0.034), and higher IFG scores (p < 0.001). The non-compliers had a greater reduction of BMI scores than the compliers (p = 0.003). Participants at risk for CVD had greater reductions than those not at risk for several risk factors, including SBP (p = 0.020), DBP (p = 0.028), IFG (p = 0.002), LDL (p = 0.006), and total serum cholesterol (p = 0.009).ConclusionWhile the physical activity intervention showed mixed results overall with both favorable changes in anthropometric indices yet unfavorable changes in plasma biomarkers, it was particularly beneficial in regards to both blood pressure indices and plasma biomarkers among those already at risk of CVD.Trial registration ClinicalTrials.gov NCT03385447; retrospectively registered

Comparison of quantification algorithms for circulating cell-free DNA methylation biomarkers in blood plasma from cancer patients

BackgroundSHOX2 and SEPT9 methylation in circulating cell-free DNA (ccfDNA) in blood are established powerful and clinically valuable biomarkers for diagnosis, staging, prognosis, and monitoring of cancer patients. The aim of the present study was to evaluate different quantification algorithms (relative quantification, absolute quantification, quasi-digital PCR) with regard to their clinical performance.MethodsMethylation analyses were performed in a training cohort (141 patients with head and neck squamous cell carcinoma [HNSCC], 170 control cases) and a testing cohort (137 HNSCC cases, 102 controls). DNA was extracted from plasma samples, bisulfite-converted, and analyzed via quantitative real-time PCR. SHOX2 and SEPT9 methylations were assessed separately and as panel [meanSEPT9/SHOX2] using the ΔCT method for absolute quantification and the ΔΔCT-method for relative quantification. Quasi-digital PCR was defined as the number of amplification-positive PCR replicates. The diagnostic (sensitivity, specificity, area under the curve (AUC) of the receiver operating characteristic (ROC)) and prognostic accuracy (hazard ratio (HR) from Cox regression) were evaluated.ResultsSporadic methylation in control samples necessitated the introduction of cutoffs resulting in 61–63% sensitivity/90–92% specificity (SEPT9/training), 53–57% sensitivity/87–90% specificity (SHOX2/training), and 64–65% sensitivity/90–91% specificity (meanSEPT9/SHOX2/training). Results were confirmed in a testing cohort with 54–56% sensitivity/88–90% specificity (SEPT9/testing), 43–48% sensitivity/93–95% specificity (SHOX2/testing), and 49–58% sensitivity/88–94% specificity (meanSEPT9/SHOX2/testing). All algorithms showed comparable cutoff-independent diagnostic accuracy with largely overlapping 95% confidence intervals (SEPT9: AUCtraining = 0.79–0.80; AUCtesting = 0.74–0.75; SHOX2: AUCtraining = 0.78–0.81, AUCtesting = 0.77–0.79; meanSEPT9/SHOX2: AUCtraining = 0.81–0.84, AUCtesting = 0.80). The accurate prediction of overall survival was possible with all three algorithms (training cohort: HRSEPT9 = 1.23-1.90, HRSHOX2 = 1.14-1.85, HRmeanSEPT9/SHOX2 =1.19-1.89 ; testing cohort: HRSEPT9 =1.22-1.67, HRSHOX2 = 1.15-1.71, HRmeanSEPT9/SHOX2 = 1.12-1.77).ConclusionThe concordant clinical performance based on different quantification algorithms allows for the application of various diagnostic platforms for the analysis of ccfDNA methylation biomarkers.

Metabolomics reveals novel blood plasma biomarkers associated to the BRCA1-mutated phenotype of human breast cancer

Hereditary breast and ovarian cancer syndrome (HBOC) is partly due to the presence of mutations in the BRCA genes. Triple-negative (TN) breast cancer (BC) shares histological characteristics with germline BRCA1 mutation-associated tumours. We have investigated the metabolic profiles of human breast cancer (BC) cell lines carrying BRCA1 pathogenic mutations by non-targeted liquid chromatography coupled to mass spectrometry technology. Based on our in vitro results, we performed a targeted metabolomic analysis of plasma samples from TN HBOC patients taking into account their BRCA1 genotype. BRCA1 promoter hypermethylation and the BRCAness phenotype of BC cell lines were also studied. The purpose of this study was to determine the metabolic signature of HBOC syndrome and TNBC patients and to evaluate the potential contribution of the metabolites identified to the genetic diagnosis of breast cancer. The present results show the existence of a differential metabolic signature for BC cells based on the BRCA1 functionality. None of the studied BC cell lines presented hypermethylation of the BRCA1 promoter region. We provide evidence of the existence of free methylated nucleotides capable of distinguishing plasma samples from HBOC patients as BRCA1-mutated and BRCA1 non-mutated, suggesting that they might be considered as BRCA1-like biomarkers for TNBC and HBOC syndrome.

Sex hormone-binding globulin (SHBG) is a potential early diagnostic biomarker for gastric cancer.

The use of blood plasma biomarkers in gastric cancer (GC) management is limited due to a lack of reliable biomarkers. An LC‐MS/MS assay and a bioinformatic analysis were performed to identify blood plasma biomarkers in a GC discovery cohort. The data obtained were verified and validated by western blotting and an ELISA in an independent study cohort. A label‐free quantification analysis of the MS data using PEAKS7 software found that four plasma proteins of apolipoprotein C‐1, gelsolin, sex hormone‐binding globulin (SHBG), and complement component C4‐A were significantly overexpressed in GC patients. A western blot assay of these plasma proteins showed that only SHBG was consistently overexpressed in the patient group. ELISA measurement of SHBG blood plasma levels confirmed that the patient group had significantly higher SHBG levels than the control group. SHBG levels in the patient group remained significantly higher after being stratified by gender, age, and disease stage. These findings show that LC‐MS/MS is powerful and highly sensitive for plasma biomarker discovery, and SHBG could be a potential plasma biomarker for GC management.

Blood plasma biomarkers of citrinin and ochratoxin A exposure in young adults in Bangladesh.

Citrinin (CIT) and Ochratoxin A (OTA) are nephrotoxic mycotoxins which can co-occur in food commodities, resulting in internal exposure. Studies in many countries reported on the presence of OTA in human blood; however, such biomonitoring data for CIT is still scarce. This study was conducted to characterize both CIT and OTA biomarker levels in plasma of volunteers since food analysis data are insufficient to assess human exposure in Bangladesh. In total 104 blood samples were collected from university students in 2013 (sampling 1: n = 64, midsummer) and 2014 (sampling 2: n = 40, end winter) for analysis of CIT and OTA and their metabolites HO-CIT and OTα by LC-MS/MS and HPLC-FD techniques, respectively. CIT and HO-CIT were detected in 90% (max 2.70ng/mL) and 85% (max 1.44ng/mL) of all samples. Mean levels in sampling 2 (CIT 0.47ng/mL; HO-CIT 0.40ng/mL) were higher than in sampling 1 (0.25ng/mL; 0.37ng/mL) indicative of variable CIT exposure. OTA was present in all (max 6.63ng/mL) and OTα in 98% (max 0.99ng/mL) of the samples. In sampling 1, mean OTA (0.85ng/mL) was higher than in sampling 2 (0.51ng/mL); the reverse situation was found for OTα mean levels. The calculated dietary OTA intake among the students (mean 9.9; max 91.7ng/kg bw/week) was lower than the tolerable weekly intake for this mycotoxin (120ng/kg bw/week) set by EFSA. But frequent co-exposure to CIT should be considered, and the results of this study indicate the necessity to identify major sources of CIT and OTA intake in the Bangladeshi population.

Blood Lead, Bone Turnover, and Survival in Amyotrophic Lateral Sclerosis.

Blood lead and bone turnover may be associated with the risk of amyotrophic lateral sclerosis (ALS). We aimed to assess whether these factors were also associated with time from ALS diagnosis to death through a survival analysis of 145 ALS patients enrolled during 2007 in the National Registry of Veterans with ALS. Associations of survival time with blood lead and plasma biomarkers of bone resorption (C-terminal telopeptides of type I collagen (CTX)) and bone formation (procollagen type I amino-terminal peptide (PINP)) were estimated using Cox models adjusted for age at diagnosis, diagnostic certainty, diagnostic delay, site of onset, and score on the Revised ALS Functional Rating Scale. Hazard ratios were calculated for each doubling of biomarker concentration. Blood lead, plasma CTX, and plasma PINP were mutually adjusted for one another. Increased lead (hazard ratio (HR) = 1.38; 95% confidence interval (CI): 1.03, 1.84) and CTX (HR = 2.03; 95% CI: 1.42, 2.89) were both associated with shorter survival, whereas higher PINP was associated with longer survival (HR = 0.59; 95% CI: 0.42, 0.83), after ALS diagnosis. No interactions were observed between lead or bone turnover and other prognostic indicators. Lead toxicity and bone metabolism may be involved in ALS pathophysiology.