Tool For Clinical Research Research Articles

Reliability, validity and responsiveness of the Dutch version of the AOSpine PROST (Patient Reported Outcome Spine Trauma)

PurposeTo validate the Dutch version of AOSpine PROST (Patient Reported Outcome Spine Trauma).MethodsPatients were recruited from two level-1 trauma centers from the Netherlands. Next to the AOSpine PROST, patients also filled out SF-36 for concurrent validity. Descriptive statistics were used to analyze the characteristics. Content validity was assessed by evaluating the number of inapplicable or missing questions. Also floor and ceiling effects were analyzed. Internal consistency was assessed by calculating Cronbach’s α and item-total correlation coefficients (itcc). Spearman correlation tests were performed within AOSpine PROST items and in correlation with SF-36. Test–retest reliability was analyzed using Intraclass Correlation Coefficients (ICC). Responsiveness was assessed by calculating effect sizes (ES) and standardized response mean (SRM). Factor analysis was performed to explore any dimensions within AOSpine PROST.ResultsOut of 179 enrolled patients, 163 (91.1%) were included. Good results were obtained for content validity. No floor or ceiling effects were seen. Internal consistency was excellent (Cronbach’s α = 0.96, itcc 0.50–0.86), with also good Spearman correlations (0.25–0.79). Compared to SF-36, the strongest correlation was seen for physical functioning (0.79; p < .001). Also test–retest reliability was excellent (ICC = 0.92). Concerning responsiveness analysis, very good results were seen with ES = 1.81 and SRM = 2.03 (p < 0.001). Factor analysis revealed two possible dimensions (Eigenvalues > 1), explaining 65.4% of variance.ConclusionsVery satisfactory results were obtained for reliability, validity and responsiveness of the Dutch version of AOSpine PROST. Treating surgeons are encouraged to use this novel and validated tool in clinical setting and research to contribute to evidence-based and patient-centered care.

High-throughput and Deep-proteome Profiling by 16-plex Tandem Mass Tag Labeling Coupled with Two-dimensional Chromatography and Mass Spectrometry.

Isobaric tandem mass tag (TMT) labeling is widely used in proteomics because of its high multiplexing capacity and deep proteome coverage. Recently, an expanded 16-plex TMT method has been introduced, which further increases the throughput of proteomic studies. In this manuscript, we present an optimized protocol for 16-plex TMT-based deep-proteome profiling, including protein sample preparation, enzymatic digestion, TMT labeling reaction, two-dimensional reverse-phase liquid chromatography (LC/LC) fractionation, tandem mass spectrometry (MS/MS), and computational data processing. The crucial quality control steps and improvements in the process specific for the 16-plex TMT analysis are highlighted. This multiplexed process offers a powerful tool for profiling a variety of complex samples such as cells, tissues, and clinical specimens. More than 10,000 proteins and posttranslational modifications such as phosphorylation, methylation, acetylation, and ubiquitination in highly complex biological samples from up to 16 different samples can be quantified in a single experiment, providing a potent tool for basic and clinical research.

Development and testing of the eye-pointing classification scale for children with cerebral palsy

Purpose The aim of this study was to develop and test a new classification scale to describe looking behaviours (gaze fixations and gaze shifts) in relation to eye-pointing. Methods The Eye-pointing Classification Scale (EpCS) was developed and tested following established procedures for the construction and evaluation of equivalent scales, and involved 2 phases: Drawing on research literature, Phase 1 involved initial drafting of the scale through a series of multi-disciplinary group discussions; evaluation of the scale through a survey procedure, and subsequent expert group evaluation. Phase 2, was an examination of scale reliability and relationships between child characteristics and level of EpCS classification. Results In Phase 1, an initial draft of the scale was developed and then evaluated by 52 participants in 10 countries, leading to its refinement. Subsequent expert evaluation of content, style and structure indicated that no further refinement was required. In Phase 2, the scale achieved excellent levels of reliability in clinical testing. A significant relationship was identified between level of child motor ability and EpCS classification, and level of child language understanding and EpCS classification. Implications for rehabilitation Non-speaking children with severe bilateral cerebral palsy who have limited upper limb movement may communicate by using controlled looking behaviours to point to objects and people, referred to as eye-pointing. However, there is little consensus as to which looking behaviours represent eye-pointing and which do not. The Eye-pointing Classification Scale (EpCS) was developed to describe looking behaviours related to eye-pointing in this population of children The EpCS provides a new robust tool for clinical management and research with children with cerebral palsy.

Polyisocyanide Hydrogels as a Tunable Platform for Mammary Gland Organoid Formation.

In the last decade, organoid technology has developed as a primary research tool in basic biological and clinical research. The reliance on poorly defined animal‐derived extracellular matrix, however, severely limits its application in regenerative and translational medicine. Here, a well‐defined, synthetic biomimetic matrix based on polyisocyanide (PIC) hydrogels that support efficient and reproducible formation of mammary gland organoids (MGOs) in vitro is presented. Only decorated with the adhesive peptide RGD for cell binding, PIC hydrogels allow MGO formation from mammary fragments or from purified single mammary epithelial cells. The cystic organoids maintain their capacity to branch for over two months, which is a fundamental and complex feature during mammary gland development. It is found that small variations in the 3D matrix give rise to large changes in the MGO: the ratio of the main cell types in the MGO is controlled by the cell–gel interactions via the cell binding peptide density, whereas gel stiffness controls colony formation efficiency, which is indicative of the progenitor density. Simple hydrogel modifications will allow for future introduction and customization of new biophysical and biochemical parameters, making the PIC platform an ideal matrix for in depth studies into organ development and for application in disease models.

Quantitative determination of agglutination based on the automatic hematology analyzer and the clinical significance of the erythrocyte-specific antibody

ObjectiveThe objective of this work was to explore the similarities and differences between the automatic hematology analyzer method and the traditional slide method in the detection of red blood cell (RBC) agglutination, and demonstrate that the automatic hematology analyzer is more intuitive and reliable for the detection of RBC agglutination. A further objective was to establish a new method to facilitate new ideas for clinical research. MethodsType A serum was selected and diluted 1:2, 1:4, 1:8, 1:16, and 1:32, to react with the type B cells and the normal saline group was used as the control group. An RBC count was performed using an automatic hematology analyzer, after incubation in a warm bath for 30 min. The degree of agglutination on the glass slide was also recorded. A positive serum of antinuclear antibody (ANA) was collected and RBC agglutination between RBC-O and ANA positive serum was determined using the automatic hematology analyzer method. ResultThe relationship between the results from the automatic hematology analyzer and the agglutination strength using the glass slide method was determined. There was a significant difference between the serum of ANA positive patients and the normal control group (P < 0.05). ConclusionA new method for detecting RBC agglutination using an automatic hematology analyzer has been established and is a valid tool for clinical research.

The plasma peptides of sepsis

BackgroundA practical strategy to discover sepsis specific proteins may be to compare the plasma peptides and proteins from patients in the intensive care unit with and without sepsis. The aim was to discover proteins and/or peptides that show greater observation frequency and/or precursor intensity in sepsis. The endogenous tryptic peptides of ICU-Sepsis were compared to ICU Control, ovarian cancer, breast cancer, female normal, sepsis, heart attack, Alzheimer’s and multiple sclerosis along with their institution-matched controls, female normals and normal samples collected directly onto ice.MethodsEndogenous tryptic peptides were extracted from individual sepsis and control EDTA plasma samples in a step gradient of acetonitrile for random and independent sampling by LC–ESI–MS/MS with a set of robust and sensitive linear quadrupole ion traps. The MS/MS spectra were fit to fully tryptic peptides within proteins using the X!TANDEM algorithm. The protein observation frequency was counted using the SEQUEST algorithm after selecting the single best charge state and peptide sequence for each MS/MS spectra. The protein observation frequency of ICU-sepsis versus ICU Control was subsequently tested by Chi square analysis. The average protein or peptide log10 precursor intensity was compared across disease and control treatments by ANOVA in the R statistical system.ResultsPeptides and/or phosphopeptides of common plasma proteins such as ITIH3, SAA2, SAA1, and FN1 showed increased observation frequency by Chi square (χ2 > 9, p < 0.003) and/or precursor intensity in sepsis. Cellular gene symbols with large Chi square values from tryptic peptides included POTEB, CTNNA1, U2SURP, KIF24, NLGN2, KSR1, GTF2H1, KIT, RPS6KL1, VAV2, HSPA7, SMC2, TCEB3B, ZNF300, SUPV3L1, ADAMTS20, LAMB4, MCCC1, SUPT6H, SCN9A, SBNO1, EPHA1, ABLIM2, cB5E3.2, EPHA10, GRIN2B, HIVEP2, CCL16, TKT, LRP2 and TMF1 amongst others showed increased observation frequency. Similarly, increased frequency of tryptic phosphopeptides were observed from POM121C, SCN8A, TMED8, NSUN7, SLX4, MADD, DNLZ, PDE3B, UTY, DEPDC7, MTX1, MYO1E, RXRB, SYDE1, FN1, PUS7L, FYCO1, USP26, ACAP2, AHI1, KSR2, LMAN1, ZNF280D and SLC8A2 amongst others. Increases in mean precursor intensity in peptides from common plasma proteins such as ITIH3, SAA2, SAA1, and FN1 as well as cellular proteins such as COL24A1, POTEB, KANK1, SDCBP2, DNAH11, ADAMTS7, MLLT1, TTC21A, TSHR, SLX4, MTCH1, and PUS7L among others were associated with sepsis. The processing of SAA1 included the cleavage of the terminal peptide D/PNHFRPAGLPEKY from the most hydrophilic point of SAA1 on the COOH side of the cystatin C binding that was most apparent in ICU-Sepsis patients compared to all other diseases and controls. Additional cleavage of SAA1 on the NH2 terminus side of the cystatin binding site were observed in ICU-Sepsis. Thus there was disease associated variation in the processing of SAA1 in ICU-Sepsis versus ICU controls or other diseases and controls.ConclusionSpecific proteins and peptides that vary between diseases might be discovered by the random and independent sampling of multiple disease and control plasma from different hospital and clinics by LC–ESI–MS/MS for storage in a relational SQL Server database and analysis with the R statistical system that will be a powerful tool for clinical research. The processing of SAA1 may play an unappreciated role in the inflammatory response to Sepsis.

A COUPLED EXPERIMENTAL AND NUMERICAL APPROACH TO CHARACTERIZE THE ANISOTROPIC MECHANICAL BEHAVIOR OF AORTIC TISSUES

Nowadays, the investigation of aortic wall biomechanics is a fundamental tool in clinical research and vascular prosthesis design. This study aims at analyzing the biomechanical behavior of aortic tissues using a coupled experimental and computational approach. Considering the typical fiber-reinforced configuration of aortic tissues, uni-axial tensile tests along six different loading directions were performed on specimens from pig aorta. Starting from the obtained experimental data, a suitable constitutive framework was defined and a methodology for the identification of the constitutive parameters was developed using the inverse analysis of mechanical tests. Transversal stretch versus loading stretch and nominal stress versus loading stretch curves were evaluated, showing the anisotropic and nonlinear mechanical behavior determined by tissue conformation with fibers distributed along preferential directions. In detail, experimental data showed different mechanical responses between longitudinal and circumferential directions, with a greater tissue stiffness along the longitudinal one. The reliability of the developed constitutive framework was evaluated by the comparison between experimental data and model results. The mentioned analysis can be considered as a useful tool for the development of reliable computational models, which allow a better understanding of the pathophysiology of cardiovascular diseases and can be applied for a proper planning of surgical procedures.

Role of RS-1 derivatives in homology-directed repair at the human genome ATG5 locus.

Genome editing is a useful tool in basic and clinical research. Among the several approaches used in genome editing, the CRISPR-Cas9 system using clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) along with a guide RNA has been developed recently. The CRISPR/Cas9 system induces site-specific double-stranded DNA breaks, which result in DNA repair via non-homologous end joining (NHEJ) or homology-directed repair (HDR). However, HDR efficiency is lower than that of NHEJ and accordingly poses a challenge in genome modification studies. Several chemical compounds including RS-1 have been shown to enhance the HDR knock-in process by two- to six-fold in HEK 293 cells and rabbit embryos. Based on this finding, we developed an antibiotic resistance system to screen RS-1 chemical derivatives, which may promote efficient HDR. In this study, we report several chemical compounds with high knock-in efficiency at the ATG5 gene locus, using HeLa cell-based assays.

4433 SGRQ score is associated with treatment status for patients with non-tuberculous mycobacterial lung disease

OBJECTIVES/GOALS: The Saint Georges Respiratory Questionnaire (SGRQ) is used as a patient reported outcome tool for clinical research in COPD and bronchiectasis. We established a registry and biospecimen repository of bronchiectasis patients with and without NTM and report associations between clinical phenotype and SGRQ scores. METHODS/STUDY POPULATION: Patients were recruited in a cross-sectional format from the Bronchiectasis, Cystic Fibrosis, and NTM clinics at our institution. All patients provided at least one sputum sample in the six months prior to inclusion. Clinical and epidemiologically relevant data was obtained, and blood specimens were processed and preserved. Patients were grouped based on clinical phenotype and differences in SGRQ scores were analyzed using ANOVA or Student’s t-test. Descriptive statistics are reported as means and standard deviations, p&lt;0.05 considered significant. RESULTS/ANTICIPATED RESULTS: 72 NTM patients completed the SGRQ including 39 patients not on treatment (Colonized), 29 patients on NTM directed antibiotics, and 4 patients whose infection was cured in the past year. Among patients on treatment, 14 were treatment refractory (positive cultures beyond 12 months of therapy). The mean age of all NTM patients was 59.5±17.6 and 80.5% were female. Mean SGRQ Total scores were significantly higher among patients receiving treatment compared to patients considered colonized (35.7± 22.0 colonized group versus 48.8± 15.8 treatment group, p = 0.011). The SGRQ subdomain scores including Impacts (26.2± 26.2 colonized group versus 42.5± 17.0 treatment group, p = 0.01) and Activities (41.7± 31.8 colonized group versus 59.3± 24.5 treatment group, p = 0.018) were also significantly different between groups. DISCUSSION/SIGNIFICANCE OF IMPACT: We developed a cross sectional cohort of NTM patients and assessed associations between clinical phenotype and SGRQ score. Preliminary data suggests that female sex, treatment status, and therapeutic duration are associated with higher SGRQ scores. We intend to continue to assess the potential for specific SGRQ questions to be used for quantifying disease symptom severity for NTM patients.

Assessment of brain volumes obtained from MP-RAGE and MP2RAGE images, quantified using different segmentation methods

For clinical purposes and research projects in neurological disease, it is of interest to evaluate the performance and comparability of available sequences and software packages for brain volume assessment to determine whether they provide equivalent results. This study compares cross-sectional brain volume values derived from images obtained with MP-RAGE or MP2RAGE sequences, using SIENA/X, SPM, or MorphoBox. MP-RAGE and MP2RAGE T1-weighted images were obtained from 24 healthy volunteers. Back-to-back scans were performed in 12 of them. Brain volumes, coefficients of variation, and concordance coefficients were determined. Significant differences were found for most brain volumes derived from MP-RAGE and MP2RAGE images. MP2RAGE-derived measures showed a non-significant trend to larger coefficients of variation. There were statistical differences between brain volumes determined with the three software packages, whereas coefficients of variation were comparable for most brain volumes. Correlation and concordance values were lower for CSF and brain parenchyma fraction measures. The results obtained advise caution when comparing brain volumes obtained by different sequences and software packages. Of note, for most brain volume measures, the MP2RAGE and MorphoBox coefficients of variation were similar to those obtained with MP-RAGE, SIENA/X or SPM, accepted tools for clinical research.

3D-Visualization of Neurovascular Compression at the Ventrolateral Medulla in Patients with Arterial Hypertension

PurposeControversy exists on the association of arterial hypertension (HTN) and neurovascular compression (NVC) at the ventrolateral medulla (VLM). No standardized and reproducible technique has been introduced yet for detection of NVC in HTN. This study aimed to generate, analyze and compare different results of exact reproducible anatomical 3D-representations of the VLM in patients with HTN, based on magnetic resonance imaging (MRI).MethodsA 3T scanner provided MRI (T2-constructive interference in steady state (CISS) high resolution imaging and three-dimensional Time-of-flight (3D-TOF) angiography) from the posterior fossa of 44 patients with clinical treatment-resistant HTN. Image processing consists of segmentation of the CISS data, registration and fusion of the CISS and TOF data and visualization. For each patient two 3D-visualizations (before and after fusion) were obtained. The reproduction quality of the vessels, flow-related signal variability and pulsation artifacts were analyzed and compared, using a ranking score.ResultsIntegrating vascular information from TOF into CISS data reduced artifacts in 3D-visualizations of exclusively processed CISS data. The quality of 3D-visualization of the vessels near the brain stem was significantly improved (p = 0.004). The results were reproducible and reliable. The quality of the 3D-presentations of neurovascular relationships at the VLM improved significantly (p < 0.001).ConclusionThe 3D-visualization of fused image data provides an excellent overview of the relationship between cranial nerves and vessels at the VLM and simplifies the detection of NVC in HTN. It provides a powerful tool for future clinical and scientific research. Although microvascular decompression (MVD) in treatment resistant HTN is not a standard procedure, it can be discussed in selected patients with intractable severe HTN.

The key points in the pre-analytical procedures of blood and urine samples in metabolomics studies.

Metabolomics provides measurement of numerous metabolites in human samples, which can be a useful tool in clinical research. Blood and urine are regarded as preferred subjects of study because of their minimally invasive collection and simple preprocessing methods. Adhering to standard operating procedures is an essential factor in ensuring excellent sample quality and reliable results. In this review, we summarize the studies about the impacts of various preprocessing factors on metabolomics studies involving clinical blood and urine samples in order to provide guidance for sample collection and preprocessing. Clinical information is important for sample grouping and data analysis which deserves attention before sample collection. Plasma and serum as well as urine samples are appropriate for metabolomics analysis. Collection tubes, hemolysis, delay at room temperature, and freeze-thaw cycles may affect metabolic profiles of blood samples. Collection time, time between sampling and examination, contamination, normalization strategies, and storage conditions may alter analysis results of urine samples. Taking these collection and preprocessing factors into account, this review provides suggestions of standard sample preprocessing.

A live cell tracking NIR bioprobe for lung cancer cell metastasis monitoring

Developing molecule imaging technology to visualize the metastasis of the cancer cells is important to facilitate an enhanced understanding of tumor biology. Staining lysosomes for the purpose of tracing cells can minimize potential interference to the cellular activities. In this study, we designed and synthesised a lysosome-targeted live cell tracking bioprobe with excellent optical properties and superior biocompatibility, which could enrich and stabilize in lysosomes to eliminate efflux after staining the cells. This bioprobe (Cy-Lyso) consists of cyanine near-infrared (NIR) fluorophore and p-fluorobenzenesulfonyl group. In the present work, p-fluorobenzenesulfonyl group was proved to be a lysosomal targeting motif independent of pH response. The cyanine dye can realize the transformation from mitochondrial targeting to lysosomal targeting after connecting with p-fluorobenzenesulfonyl, and thereby long term fluorescence in living cells. This probe was utilized for sustained fluorescence labeling of living cells, successful located the position of lung cancer cells A549 in co-culture status. We further utilized this probe to label A549 cells and observe the metastasis way of A549 in vivo, and revealed that the liver and intestine are the main metastasis organs of A549 cells. In summary, this study provides an effective tool for basic research and clinical research on single cell behavior.

Use of Immersive Virtual Reality in the Assessment and Treatment of Alzheimer's Disease: A Systematic Review.

Background:Immersive virtual reality (iVR) allows seamless interaction with simulated environments and is becoming an established tool in clinical research. It is unclear whether iVR is acceptable to people with Alzheimer’s disease (AD) dementia or useful in their care. We explore whether iVR is a viable research tool that may aid the detection and treatment of AD.Objectives:This review examines the use of iVR in people with AD or mild cognitive impairment (MCI).Methods:Medline, PsycINFO, Embase, CINAHL, and Web of Science databases were searched from inception. PRISMA guidelines were used with studies selected by at least two researchers.Results:Nine studies were eligible for inclusion. None reported any issues with iVR tolerability in participants with MCI and AD on assessment or treatment tasks. One study demonstrated capability for detecting prodromal AD and correlated with neuroanatomical substrates. Two studies showed iVR to have high accuracy in differentiating participants with AD from controls but were not hypothesis driven or with adequate controls measures. In a small validation study and two longitudinal case studies, iVR cognitive training was positively rated but did not demonstrate reliable benefit.Conclusion:iVR is emerging as a viable method of assessing older adults and people with AD. Strongest benefits were seen when closely integrated with theoretical models of neurodegeneration and existing screening methods. Further randomized controlled trials integrated with clinical populations are required. This will consolidate the power of iVR for assessment of MCI and clarify treatment efficacy beyond current applications in physical rehabilitation.

A Novel, Reliable Protocol to Objectively Assess Scar Stiffness Using Shear Wave Elastography

The aim of this research was to investigate the use of shear wave elastography as a novel tool to quantify and visualize scar stiffness after a burn. Increased scar stiffness is indicative of pathologic scarring which is associated with persistent pain, chronic itch and restricted range of movement. Fifty-five participants with a total of 96 scars and 69 contralateral normal skin sites were evaluated. A unique protocol was developed to enable imaging of the raised and uneven burn scars. Intra-rater and inter-rater reliability was excellent (intra-class correlation coefficient >0.97), and test–retest reliability was good (intra-class correlation coefficient >0.85). Shear wave elastography was able to differentiate between normal skin, pathologic scars and non-pathologic scars, with preliminary cutoff values identified. Significant correlations were found between shear wave velocity and subjective clinical scar assessment (r = 0.66). Shear wave elastography was able to provide unique information associated with pathologic scarring and shows promise as a clinical assessment and research tool.

Coronavirus Anxiety Scale: A brief mental health screener for COVID-19 related anxiety

Mental health concerns of people impacted by the coronavirus pandemic have not been adequately addressed. The objective of this study was to develop and evaluate the properties of the Coronavirus Anxiety Scale (CAS), which is a brief mental health screener to identify probable cases of dysfunctional anxiety associated with the COVID-19 crisis. This 5-item scale, which was based on 775 adults with anxiety over the coronavirus, demonstrated solid reliability and validity. Elevated CAS scores were found to be associated with coronavirus diagnosis, impairment, alcohol/drug coping, negative religious coping, extreme hopelessness, suicidal ideation, as well as attitudes toward President Trump and Chinese products. The CAS discriminates well between persons with and without dysfunctional anxiety using an optimized cut score of ≥ 9 (90% sensitivity and 85% specificity). These results support the CAS as an efficient and valid tool for clinical research and practice.

Holographic virtual staining of individual biological cells

Many medical and biological protocols for analyzing individual biological cells involve morphological evaluation based on cell staining, designed to enhance imaging contrast and enable clinicians and biologists to differentiate between various cell organelles. However, cell staining is not always allowed in certain medical procedures. In other cases, staining may be time-consuming or expensive to implement. Staining protocols may be operator-sensitive, and hence may lead to varying analytical results, as well as cause artificial imaging artifacts or false heterogeneity. We present a deep-learning approach, called HoloStain, which converts images of isolated biological cells acquired without staining by holographic microscopy to their virtually stained images. We demonstrate this approach for human sperm cells, as there is a well-established protocol and global standardization for characterizing the morphology of stained human sperm cells for fertility evaluation, but, on the other hand, staining might be cytotoxic and thus is not allowed during human in vitro fertilization (IVF). After a training process, the deep neural network can take images of unseen sperm cells retrieved from holograms acquired without staining and convert them to their stainlike images. We obtained a fivefold recall improvement in the analysis results, demonstrating the advantage of using virtual staining for sperm cell analysis. With the introduction of simple holographic imaging methods in clinical settings, the proposed method has a great potential to become a common practice in human IVF procedures, as well as to significantly simplify and radically change other cell analyses and techniques such as imaging flow cytometry.

An LC/MS/MS method for analyzing the steroid metabolome with high accuracy and from small serum samples

Analyzing global steroid metabolism in humans can shed light on the etiologies of steroid-related diseases. However, existing methods require large amounts of serum and lack the evaluation of accuracy. Here, we developed an LC/MS/MS method for the simultaneous quantification of 12 steroid hormones: testosterone, pregnenolone, progesterone, androstenedione, corticosterone, 11-deoxycortisol, cortisol, 17-hydroxypregnenolone, 17-hydroxyprogesterone, dehydroepiandrosterone, estriol, and estradiol. Steroids and spiked internal standards in 100 μl serum were extracted by protein precipitation and liquid-liquid extraction. The organic phase was dried by evaporation, and isonicotinoyl chloride was added for steroid derivatization, followed by evaporation under nitrogen and redissolution in 50% methanol. Chromatographic separation was performed on a reverse-phase PFP column, and analytes were detected on a triple quadrupole mass spectrometer with ESI. The lower limits of quantification ranged from 0.005 ng/ml for estradiol to 1 ng/ml for cortisol. Apparent recoveries of steroids at high, medium, and low concentrations in quality control samples were between 86.4% and 115.0%. There were limited biases (−10.7% to 10.5%) between the measured values and the authentic values, indicating that the method has excellent reliability. An analysis of the steroid metabolome in pregnant women highlighted the applicability of the method in clinical serum samples. We conclude that the LC/MS/MS method reported here enables steroid metabolome analysis with high accuracy and reduced serum consumption, indicating that it may be a useful tool in both clinical and scientific laboratory research.

The TOPAS tool for particle simulation, a Monte Carlo simulation tool for physics, biology and clinical research

PurposeThis paper covers recent developments and applications of the TOPAS TOol for PArticle Simulation and presents the approaches used to disseminate TOPAS. Materials and methodsFundamental understanding of radiotherapy and imaging is greatly facilitated through accurate and detailed simulation of the passage of ionizing radiation through apparatus and into a patient using Monte Carlo (MC). TOPAS brings Geant4, a reliable, experimentally validated MC tool mainly developed for high energy physics, within easy reach of medical physicists, radiobiologists and clinicians. Requiring no programming knowledge, TOPAS provides all of the flexibility of Geant4. ResultsAfter 5 years of development followed by its initial release, TOPAS was subsequently expanded from its focus on proton therapy physics to incorporate radiobiology modeling. Next, in 2018, the developers expanded their user support and code maintenance as well as the scope of TOPAS towards supporting X-ray and electron therapy and medical imaging. Improvements have been achieved in user enhancement through software engineering and a graphical user interface, calculational efficiency, validation through experimental benchmarks and QA measurements, and either newly available or recently published applications. A large and rapidly increasing user base demonstrates success in our approach to dissemination of this uniquely accessible and flexible MC research tool. ConclusionsThe TOPAS developers continue to make strides in addressing the needs of the medical community in applications of ionizing radiation to medicine, creating the only fully integrated platform for four-dimensional simulation of all forms of radiotherapy and imaging with ionizing radiation, with a design that promotes inter-institutional collaboration.

Rapid Prostate Cancer Noninvasive Biomarker Screening Using Segmented Flow Mass Spectrometry-Based Untargeted Metabolomics.

Spectrometric methods with rapid biomarker detection capacity through untargeted metabolomics are becoming essential in the clinical cancer research. Liquid chromatography-mass spectrometry (LC-MS) is a rapidly developing metabolomic-based biomarker technique due to its high sensitivity, reproducibility, and separation efficiency. However, its translation to clinical diagnostics is often limited due to long data acquisition times (∼20 min/sample) and laborious sample extraction procedures when employed for large-scale metabolomics studies. Here, we developed a segmented flow approach coupled with high-resolution mass spectrometry (SF-HRMS) for untargeted metabolomics, which has the capability to acquire data in less than 1.5 min/sample with robustness and reproducibility relative to LC-HRMS. The SF-HRMS results demonstrate the capability for screening metabolite-based urinary biomarkers associated with prostate cancer (PCa). The study shows that SF-HRMS-based global metabolomics has the potential to evolve into a rapid biomarker screening tool for clinical research.