Non-invasive Methodology Research Articles

P–240 Human extracellular vesicles (EVs) secreted by aneuploid embryos potentiate development of non-invasive PGT-A RNA biomarkers and stimulate MUC1 up-regulation in primary endometrial stromal cells (ESCs)

Abstract Study question Could EVs secreted by aneuploid embryos a) serve for development of RNA biomarkers for PGT-A and b) elicit a relevant transcriptomic response in decidualised ESCs? Summary answer Aneuploid embryo EVs a) contain PPM1J, LINC00561, ANKRD34C and TMED10 in differential abundance from euploid EVs and b) induce up-regulation of MUC1 in decidualised ESCs. What is known already Embryo aneuploidy accounts for approximately 50% of all recurrent implantation failures in women >35 years old. PGT-A identifies euploid embryos to increase implantation probability but the technology is controversial as it requires an invasive embryo biopsy with an elusive long-term biosafety. The development of non-invasive methods to screen out aneuploid embryos is paramount. It is also critical to decode the embryo-endometrial dialog underlying implantation failure. We have previously reported that IVF embryos secrete EVs that can be internalised by ESCs, conceptualising that successful implantation to the endometrium is facilitated by EVs, which may additionally serve as biomarkers of ploidy status. Study design, size, duration Embryos destined for biopsy on days 5–7 for PGT-A were grown under standard conditions. Spent media (30μl) were collected from euploid (n = 175) and aneuploid embryos (n = 145) at both cleavage (days 1–3) and blastocyst (days 3–5) stage. Media samples from n = 35 cleavage embryos were pooled in order to obtain five euploid and four aneuploidy pools. Blastocyst media were pooled to create one euploid and one aneuploid pool. ESCs were obtained from five women undergoing diagnostic laparoscopy. Participants/materials, setting, methods The study was realised at a research hospital. EVs were isolated from euploid and aneuploid Day3 pools with differential ultracentrifugation and EV-RNA sequencing was performed following the SMARTer Stranded Total RNA-Seq approach. ESCs were decidualised (E2:10nM, P4:1uM, cAMP:0.5 mM twice every 48 hours) and treated for 24 hours with 50 ng/ml euploid or aneuploid EVs extracted from blastocyst media. RNA sequencing was performed on ESCs following the Truseq RNAseq protocol. Main results and the role of chance Aneuploid cleavage stage embryos (n = 4) secreted EVs that were less abundant in RNA fragments originating from the genes PPM1J (log2fc=–5.13, p = 0.011), LINC00561 (log2fc=–7.87, p = 0.010) and ANKRD34C (log2fc=–7.30, p = 0.017) and more abundant in TMED10 (log2fc=1.63 p = 0.025) compared to EVs (n = 5) from euploid embryos. Decidualisation per se induced downregulation of MUC1 (log2FC=–0.54, p = 0.0028) in ESCs as prerequisite for the establishment of receptive endometrium. The expression of MUC1 transcript in decidualised ESCs was significantly increased following treatment with aneuploid compared to euploid embryo-secreted EVs (log2FC=0.85, p = 0.0201). Limitations, reasons for caution The findings of the study may require validation utilising a second cohort of EVs samples. Wider implications of the findings: This discovery that the RNA cargo of EVs secreted from aneuploid cleavage stage embryos is diverse from that of euploid embryos potentiates the development of non-invasive methodology for PGT-A. The upregulation of MUC1 in decidualised ESCs following aneuploid embryo EV treatment proposes a new mechanism underlying implantation failure. Trial registration number NA

P-240 Human extracellular vesicles (EVs) secreted by aneuploid embryos potentiate development of non-invasive PGT-A RNA biomarkers and stimulate MUC1 up-regulation in primary endometrial stromal cells (ESCs)

Abstract Study question Could EVs secreted by aneuploid embryos a) serve for development of RNA biomarkers for PGT-A and b) elicit a relevant transcriptomic response in decidualised ESCs? Summary answer Aneuploid embryo EVs a) contain PPM1J, LINC00561, ANKRD34C and TMED10 in differential abundance from euploid EVs and b) induce up-regulation of MUC1 in decidualised ESCs. What is known already Embryo aneuploidy accounts for approximately 50% of all recurrent implantation failures in women >35 years old. PGT-A identifies euploid embryos to increase implantation probability but the technology is controversial as it requires an invasive embryo biopsy with an elusive long-term biosafety. The development of non-invasive methods to screen out aneuploid embryos is paramount. It is also critical to decode the embryo-endometrial dialog underlying implantation failure. We have previously reported that IVF embryos secrete EVs that can be internalised by ESCs, conceptualising that successful implantation to the endometrium is facilitated by EVs, which may additionally serve as biomarkers of ploidy status. Study design, size, duration Embryos destined for biopsy on days 5-7 for PGT-A were grown under standard conditions. Spent media (30μl) were collected from euploid (n = 175) and aneuploid embryos (n = 145) at both cleavage (days 1-3) and blastocyst (days 3-5) stage. Media samples from n = 35 cleavage embryos were pooled in order to obtain five euploid and four aneuploidy pools. Blastocyst media were pooled to create one euploid and one aneuploid pool. ESCs were obtained from five women undergoing diagnostic laparoscopy. Participants/materials, setting, methods The study was realised at a research hospital. EVs were isolated from euploid and aneuploid Day3 pools with differential ultracentrifugation and EV-RNA sequencing was performed following the SMARTer Stranded Total RNA-Seq approach. ESCs were decidualised (E2:10nM, P4:1uM, cAMP:0.5 mM twice every 48 hours) and treated for 24 hours with 50 ng/ml euploid or aneuploid EVs extracted from blastocyst media. RNA sequencing was performed on ESCs following the Truseq RNAseq protocol. Main results and the role of chance Aneuploid cleavage stage embryos (n = 4) secreted EVs that were less abundant in RNA fragments originating from the genes PPM1J (log2fc=-5.13, p = 0.011), LINC00561 (log2fc=-7.87, p = 0.010) and ANKRD34C (log2fc=-7.30, p = 0.017) and more abundant in TMED10 (log2fc=1.63 p = 0.025) compared to EVs (n = 5) from euploid embryos. Decidualisation per se induced downregulation of MUC1 (log2FC=-0.54, p = 0.0028) in ESCs as prerequisite for the establishment of receptive endometrium. The expression of MUC1 transcript in decidualised ESCs was significantly increased following treatment with aneuploid compared to euploid embryo-secreted EVs (log2FC=0.85, p = 0.0201). Limitations, reasons for caution The findings of the study may require validation utilising a second cohort of EVs samples. Wider implications of the findings This discovery that the RNA cargo of EVs secreted from aneuploid cleavage stage embryos is diverse from that of euploid embryos potentiates the development of non-invasive methodology for PGT-A. The upregulation of MUC1 in decidualised ESCs following aneuploid embryo EV treatment proposes a new mechanism underlying implantation failure. Trial registration number NA

Not All Lesioned Tissue Is Equal: Identifying Pericavitational Areas in Chronic Stroke With Tissue Integrity Gradation via T2w T1w Ratio.

Stroke-related tissue damage within lesioned brain areas is topologically non-uniform and has underlying tissue composition changes that may have important implications for rehabilitation. However, we know of no uniformly accepted, objective non-invasive methodology to identify pericavitational areas within the chronic stroke lesion. To fill this gap, we propose a novel magnetic resonance imaging (MRI) methodology to objectively quantify the lesion core and surrounding pericavitational perimeter, which we call tissue integrity gradation via T2w T1w ratio (TIGR). TIGR uses standard T1-weighted (T1w) and T2-weighted (T2w) anatomical images routinely collected in the clinical setting. TIGR maps are analyzed with relation to subject-specific gray matter and cerebrospinal fluid thresholds and binned to create a false colormap of tissue damage within the stroke lesion, and these are further categorized into low-, medium-, and high-damage areas. We validate TIGR by showing that the cerebral blood flow within the lesion reduces with greater tissue damage (p = 0.005). We further show that a significant task activity can be detected in pericavitational areas and that medium-damage areas contain a significantly lower magnitude of hemodynamic response function than the adjacent damaged areas (p < 0.0001). We also demonstrate the feasibility of using TIGR maps to extract multivariate brain–behavior relationships (p < 0.05) and show general agreement in location compared to binary lesion, T1w-only, and T2w-only maps but that the extent of brain behavior maps may depend on signal sensitivity as denoted by the sparseness coefficient (p < 0.0001). Finally, we show the feasibility of quantifying TIGR in early and late subacute stroke phases, where higher-damage areas were smaller in size (p = 0.002) and that lesioned voxels transition from lower to higher damage with increasing time post-stroke (p = 0.004). We conclude that TIGR is able to (1) identify tissue damage gradient within the stroke lesion across different post-stroke timepoints and (2) more objectively delineate lesion core from pericavitational areas wherein such areas demonstrate reasonable and expected physiological and functional impairments. Importantly, because T1w and T2w scans are routinely collected in the clinic, TIGR maps can be readily incorporated in clinical settings without additional imaging costs or patient burden to facilitate decision processes related to rehabilitation planning.

Enabling Wearable Pulse Transit Time-Based Blood Pressure Estimation for Medically Underserved Areas and Health Equity: Comprehensive Evaluation Study.

BackgroundNoninvasive and cuffless approaches to monitor blood pressure (BP), in light of their convenience and accuracy, have paved the way toward remote screening and management of hypertension. However, existing noninvasive methodologies, which operate on mechanical, electrical, and optical sensing modalities, have not been thoroughly evaluated in demographically and racially diverse populations. Thus, the potential accuracy of these technologies in populations where they could have the greatest impact has not been sufficiently addressed. This presents challenges in clinical translation due to concerns about perpetuating existing health disparities.ObjectiveIn this paper, we aim to present findings on the feasibility of a cuffless, wrist-worn, pulse transit time (PTT)–based device for monitoring BP in a diverse population.MethodsWe recruited a diverse population through a collaborative effort with a nonprofit organization working with medically underserved areas in Georgia. We used our custom, multimodal, wrist-worn device to measure the PTT through seismocardiography, as the proximal timing reference, and photoplethysmography, as the distal timing reference. In addition, we created a novel data-driven beat-selection algorithm to reduce noise and improve the robustness of the method. We compared the wearable PTT measurements with those from a finger-cuff continuous BP device over the course of several perturbations used to modulate BP.ResultsOur PTT-based wrist-worn device accurately monitored diastolic blood pressure (DBP) and mean arterial pressure (MAP) in a diverse population (N=44 participants) with a mean absolute difference of 2.90 mm Hg and 3.39 mm Hg for DBP and MAP, respectively, after calibration. Meanwhile, the mean absolute difference of our systolic BP estimation was 5.36 mm Hg, a grade B classification based on the Institute for Electronics and Electrical Engineers standard. We have further demonstrated the ability of our device to capture the commonly observed demographic differences in underlying arterial stiffness.ConclusionsAccurate DBP and MAP estimation, along with grade B systolic BP estimation, using a convenient wearable device can empower users and facilitate remote BP monitoring in medically underserved areas, thus providing widespread hypertension screening and management for health equity.

Biodistribution of 68/67Ga-Radiolabeled Sphingolipid Nanoemulsions by PET and SPECT Imaging.

Background and PurposeNon-invasive imaging methodologies, especially nuclear imaging techniques, have undergone an extraordinary development over the last years. Interest in the development of innovative tracers has prompted the emergence of new nanomaterials with a focus on nuclear imaging and therapeutical applications. Among others, organic nanoparticles are of the highest interest due to their translational potential related to their biocompatibility and biodegradability. Our group has developed a promising new type of biocompatible nanomaterials, sphingomyelin nanoemulsions (SNs). The aim of this study is to explore the potential of SNs for nuclear imaging applications.MethodsReady-to-label SNs were prepared by a one-step method using lipid derivative chelators and characterized in terms of their physicochemical properties. Stability was assessed under storage and after incubation with human serum. Chelator-functionalized SNs were radiolabeled with 67Ga and 68Ga, and the radiochemical yield (RCY), radiochemical purity (RCP) and radiochemical stability (RCS) were determined. Finally, the biodistribution of 67/68Ga-SNs was evaluated in vivo and ex vivo.ResultsHere, we describe a simple and mild one-step method for fast and efficient radiolabeling of SNs with 68Ga and 67Ga radioisotopes. In vivo experiments showed that 67/68Ga-SNs can efficiently and indistinctly be followed up by PET and SPECT. Additionally, we proved that the biodistribution of the 67/68Ga-SNs can be conveniently modulated by modifying the surface properties of different hydrophilic polymers, and therefore the formulation can be further adapted to the specific requirements of different biomedical applications.ConclusionThis work supports 67/68Ga-SNs as a novel probe for nuclear imaging with tunable biodistribution and with great potential for the future development of nanotheranostics.

A global non-invasive methodology for the phenotyping of potato under water deficit conditions using imaging, physiological and molecular tools

BackgroundDrought is a major consequence of global heating that has negative impacts on agriculture. Potato is a drought-sensitive crop; tuber growth and dry matter content may both be impacted. Moreover, water deficit can induce physiological disorders such as glassy tubers and internal rust spots. The response of potato plants to drought is complex and can be affected by cultivar type, climatic and soil conditions, and the point at which water stress occurs during growth. The characterization of adaptive responses in plants presents a major phenotyping challenge. There is therefore a demand for the development of non-invasive analytical techniques to improve phenotyping.ResultsThis project aimed to take advantage of innovative approaches in MRI, phenotyping and molecular biology to evaluate the effects of water stress on potato plants during growth. Plants were cultivated in pots under different water conditions. A control group of plants were cultivated under optimal water uptake conditions. Other groups were cultivated under mild and severe water deficiency conditions (40 and 20% of field capacity, respectively) applied at different tuber growth phases (initiation, filling). Water stress was evaluated by monitoring soil water potential. Two fully-equipped imaging cabinets were set up to characterize plant morphology using high definition color cameras (top and side views) and to measure plant stress using RGB cameras. The response of potato plants to water stress depended on the intensity and duration of the stress. Three-dimensional morphological images of the underground organs of potato plants in pots were recorded using a 1.5 T MRI scanner. A significant difference in growth kinetics was observed at the early growth stages between the control and stressed plants. Quantitative PCR analysis was carried out at molecular level on the expression patterns of selected drought-responsive genes. Variations in stress levels were seen to modulate ABA and drought-responsive ABA-dependent and ABA-independent genes.ConclusionsThis methodology, when applied to the phenotyping of potato under water deficit conditions, provides a quantitative analysis of leaves and tubers properties at microstructural and molecular levels. The approaches thus developed could therefore be effective in the multi-scale characterization of plant response to water stress, from organ development to gene expression.

Development of convolutional neural networks for recognition of tenogenic differentiation based on cellular morphology

Background and objective: The use of automated systems for image recognition is highly preferred for regenerative medicine applications to evaluate stem cell differentiation early in the culturing state with non-invasive methodologies instead of invasive counterparts. Bone marrow-derived mesenchymal stem cells (BMSCs) are able to differentiate into desired cell phenotypes, and thereby promise a proper cell source for tendon regeneration. The therapeutic success of stem cell therapy requires cellular characterization prior to the implantation of cells. The foremost problem is that traditional characterization techniques require cellular material which would be more useful for cell therapy, complex laboratory procedures, and human expertise. Convolutional neural networks (CNNs), a class of deep neural networks, have recently made great improvements in image-based classifications, recognition, and detection tasks. We, therefore, aim to develop a potential CNN model in order to recognize differentiated stem cells by learning features directly from image data of unlabelled cells.Methods: The differentiation of bone marrow mesenchymal stem cells (BMSCs) into tenocytes was induced with the treatment of bone morphogenetic protein-12 (BMP-12). Following the treatment and incubation step, the phase-contrast images of cells were obtained and immunofluorescence staining has been applied to characterize the differentiated state of BMSCs. CNN models were developed and trained with the phase-contrast cell images. The comparison of CNN models was performed with respect to prediction performance and training time. Moreover, we have evaluated the effect of image enhancement method, data augmentation, and fine-tuning training strategy to increase classification accuracy of CNN models. The best model was integrated into a mobile application.Results: All the CNN models can fit the biological data extracted from immunofluorescence characterization. CNN models enable the cell classification with satisfactory accuracies. The best result in terms of accuracy and training time is achieved by the model proposed based on Inception-ResNet V2 trained from scratch using image enhancement and data augmentation strategies (96.80%, 434.55 sec).Conclusion: Our study reveals that the CNN models show good performance by identifying stem cell differentiation. Importantly this technique provides a faster and real-time tool in comparison to traditional methods enabling the adjustment of culture conditions during cultivation to improve the yield of therapeutic stem cells.

Raman Spectroscopy Technique: A Non-Invasive Tool in Celiac Disease Diagnosis.

Celiac disease (CD) is diagnosed by a combination of specific serology and typical duodenal lesions. The histological confirmation of CD, mandatory in the majority of patients with suspected CD, is based on invasive and poorly tolerated procedures, such as upper gastrointestinal endoscopy. In this study we propose an alternative and non-invasive methodology able to confirm the diagnosis of CD based on the analysis of serum samples using the Raman spectroscopy technique. Three different bands centered at 1650, 1450 and 1003 cm−1 have been considered and the A1450/A1003 and A1650/A1003 ratios have been computed to discriminate between CD and non-CD subjects. The reliability of the methodology was validated by statistical analysis using receiver operating characteristic (ROC) curves. The Youden index was also determined to obtain optimal cut-off points. The obtained results highlighted that the proposed methodology was able to distinguish between CD and non-CD subjects with 98% accuracy. The optimal cut-off points revealed, for both the A1450/A1003 and A1650/A1003 ratios, high values of sensitivity and specificity (>95.0% and >92.0% respectively), confirming that Raman spectroscopy may be considered a valid alternative to duodenal biopsy and demonstrates spectral changes in the secondary structures of the protein network.

Non-Invasive Technical Investigation of English Portrait Miniatures Attributed to Nicholas Hilliard and Isaac Oliver

This study presents the results of the technical investigation carried out on several English portrait miniatures painted in the 16th and 17th century by Nicholas Hilliard and Isaac Oliver, two of the most famous limners working at the Tudor and Stuart courts. The 23 objects chosen for the analysis, spanning almost the entire career of the two artists, belong to the collections of the Victoria and Albert Museum (London) and the Fitzwilliam Museum (Cambridge). A non-invasive scientific methodology, comprising of stereo and optical microscopies, Raman microscopy, and X-ray fluorescence spectroscopy, was required for the investigation of these small-scale and fragile objects. The palettes and working techniques of the two artists were characterised, focusing in particular on the examination of flesh tones, mouths, and eyes. These findings were also compared to the information written in the treatises on miniature painting circulating during the artists’ lifetime. By identifying the materials and techniques most widely employed by the two artists, this study provides information about similarities and differences in their working methods, which can help to understand their artistic practice as well as contribute to matters of attribution.

Developing non-invasive bladder cancer screening methodology through potentiometric multisensor urine analysis

We report on the feasibility study exploring the potential of a simple electrochemical multisensor system as a tool for distinguishing between urine samples from patients with confirmed bladder cancer (36 samples) and healthy volunteers (51 samples). The potentiometric sensor responses obtained in urine samples were employed as the input data for various machine learning classification algorithms (logistic regression, random forest, extreme gradient boosting classifier, support vector machine, and voting classifier). The performance metrics of the classifiers were evaluated via Monte-Carlo cross-validation. The best model combining all the acquired data from the people aged 19–88 with different tumor grades and malignancies, including patients with recurrent bladder cancer, yielded 72% accuracy, 71% sensitivity, and 58% specificity. It was found that these metrics can be improved to 76% accuracy, 80% sensitivity, and 75% specificity when only a limited age group (50–88 years of age) is considered. Taking into account the simplicity of the proposed screening method, this technique appears to be a promising tool for further research.

Infrared thermography: A new approach for the characterization and management of activated carbons applied in water treatment

A new non-invasive methodology based on infrared thermography (IRT) to evaluate the exhaustion level of granular activated carbon (GAC) at different depths of a target industrial water treatment filter is studied. IRT results are discussed in terms of adsorption capacity and correlated with dedicated analytical methods: N2 gas adsorption, TGA, TD-GC/MS and NMR. Findings revealed that the surface temperature and surface thermal density parameters are contingent with the exhaustion level as well as the surface area and pore volumes of the GAC samples. Surface temperature decreased with 80% and the surface thermal density with 35% comparing exhausted Top-GAC and virgin GAC. Linear correlation was found between IRT parameters and the exhaustion profile of 98%. During GAC exploitation the surface area was reduced to 85% and the total volume of pore to 80%. The IRT method demonstrated to be simple and accurate and suitable for GAC management of water treatment.

Medical Parameters of a Sensing Device for Colorectal Cancer Preventive Screening through Fecal Odor Analysis

Colorectal cancer (CRC) is a worldwide diffused pathology for both men and women and it is the third most common cancer for both genders, in the United States[1] However, if promptly diagnosed, CRC is also one of the most curable tumor types (90% curability rate at stage I), therefore prevention is fundamental to avoid degeneration. Here the clinical validation outcomes of a preventive screening method based on a patented device, SCENT A1, composed by a core of nanostructured MOX sensors [2]. SCENT A1 is capable of detecting the presence of CRC from fecal odor with an in-vitro, inexpensive and non-invasive methodology. Fecal odor can be affected by the presence of CRC-biomarkers produced by membrane peroxidation and metabolic alterations [3,4]. The screening analysis adopted by NHS of Italy and of other countries is fecal occult blood test (FOBT). In Italy, FOBT is performed every two years on subjects from 50 to 69 years old and shows a huge percentage of false positives (about 65% according to data). FOBT-positives are then invited to undergo colonoscopy. SCENT A1 is capable of reducing FOBT false positives and so the total number of non-operative colonoscopies with related risks (e.g. bowel perforation). The device, presently undergoing instrumental certification, is composed by a core of two chemoresistive MOX sensors, chosen and calibrated for this specific aim [5-11]. Sensors (Figure 1) have been synthesized at Sensor Laboratory of the University of Ferrara and are composed by a nanostructured semiconductor film screen-printed onto an alumina substrate and a platinum heater to modulate operating temperature. The first sensor is composed by Iron and Samarium oxides while the second one by Tin and Titanium oxides. The data collection and analysis software employs support vector machine (SVM) [9]. The clinical validation protocol of this device started in May 2016 and ended in July 2019, in collaboration with Hospital St. Anna of Ferrara and Department of Public Health of Ferrara. A total of 398 fecal samples of FOBT-positive subjects have been analyzed by SCENT A1 system with the k-fold cross validation method. Samples have been grouped into two macro categories, depending on the gold-standard (colonoscopy) outcomes: 260 healthy subjects (negative to colonoscopy); 138 colorectal adenomas and carcinomas (positive to colonoscopy), 54 low-risk adenomas and 84 high-risk adenomas and carcinomas. The system has been capable of distinguishing among these two categories with a sensitivity (TPR: true positive rate) and specificity (TNR: true negative rate) respectively of 84,1% and 82,4%, defined as follows: TPR=TP/P=TP/(FN+TP);TN/N=TN/(FP+TN); where TP, TN, FP and FN are the abbreviations that indicate respectively true positives, true negatives, false positives and false negatives. The choice to group carcinomas, high- and low-risk adenomas into a single class and not into two diverse classes as before [10], has simplified the test and ensured greater doctor protection, despite a small loss in specificity. Low-risk adenomas are, in fact, often similar to healthy tissues and have a risk of evolving into cancer comparable to the risk of colonoscopy complications. As already emerging from a previous study [10-11], in fact, the major error of classification (only 57%) concerned just low-risk tumors that emit gases halfway between healthy and tumoral samples. What emerges is that, if SCENT A1 test is carried out on FOBT-positives subjects as a second check, it would guarantee a reduction of about two thirds of the colonoscopies performed on healthy patients. A new collaboration, involving private clinic Quisisana, started in 2020 with some laboratory tests on sensors reproducibility [12] will continue in January 2021 with tests on patients directly inside the clinic, in order to test the new shape of the device under certification. Moreover, if employed by the NHS of countries without screening system, this method will significantly reduce CRC-mortality rate.[1] [https://www.cancer.org/cancer/colon-rectal-cancer/about/key-statistics.html [2] Italian #: RM2014A000595, European #: 3210013 (Germany, UK); [3] B. Szachowicz-Petelska et al., NMR in Biomedicine V5,226–233, 1992; [4] T. G. de Meij et al., International Journal of Cancer (2014), 134, 1132–1138; [5] C. Malagù et al., Sensors, 14, 18982-18992, 2014; [6] G. Zonta et al., Sensors and Actuators B, 218, 289-295, 2015; [7] N. Landini et al., Scholars' Press, ISBN-13: 978-3-639-76538-0; [8] G. Zonta et al., Sensors and Actuators B, 238, 1098–110, 2016; [9] G. Zonta et al., Sensors and Actuators B 262 (2018) 884–891; [10] G. Zonta et al., Sensors and Actuators B: Chemical, Volume 301, 12 December 2019, 127062;[11] G. Zonta et al., Cancers 2020, 12(6), 1471; [12] G. Zonta et al., Ceramics International Volume 46, Issue 5, 1 April 2020, Pages 6847-6855.Figure 1: a) external view of the patented prototype SCENT A1, composed by flowmeters, a flow diverter and a sample box; b) one of the chemoresistive sensors composing the core of the instrument; c) magnificated scheme of the sensor. Figure 1

Investigation of the cyclic aging characteristics and degradation mechanism of LiNi1/3Co1/3Mn1/3O2 batteries by non-invasive methodologies

Cyclic aging tests of LiNi1/3Co1/3Mn1/3O2 batteries are performed at 25℃, 45℃ and 60℃. Several non-invasive electrochemical techniques, such as incremental capacity analysis, differential voltage analysis and capacity difference analysis, are employed to deduce degradation mechanism. The degradation mechanism of batteries is similar at different temperatures. Loss of active positive material follows a linear function at all conditions and increases by elevating temperature. Due to the graphite's volume expansion and contraction, loss of active negative material is observed at the first 100 full cycle equivalents. Then, it disappears until the appearance of a dense lithium-containing passivation layer. This covering layer is mainly from lithium deposition on the negative electrode surface. With this lithium-impermeable covering layer's growth, the homogeneity of active lithium distribution is successively reduced, and loss of negative material increases gradually. Moreover, a worst cyclic performance is found at 45℃, where the homogeneity of electrodes reduces rapidly. All results reveal that loss of active materials and lithium inventory loss only influence capacity loss. The evolution of the covering layer on the negative electrode surface is the deciding factor for cycle life.

Biological autoluminescence for assessing oxidative processes in yeast cell cultures

Nowadays, modern medicine is looking for new, more gentle, and more efficient diagnostic methods. A pathological state of an organism is often closely connected with increased amount of reactive oxygen species. They can react with biomolecules and subsequent reactions can lead to very low endogenous light emission (biological autoluminescence—BAL). This phenomenon can be potentially used as a non-invasive and low-operational-cost tool for monitoring oxidative stress during diseases. To contribute to the understanding of the parameters affecting BAL, we analyzed the BAL from yeast Saccharomyces cerevisiae as a representative eukaryotic organism. The relationship between the BAL intensity and the amount of reactive oxygen species that originates as a result of the Fenton reaction as well as correlation between spontaneous BAL and selected physical and chemical parameters (pH, oxygen partial pressure, and cell concentration) during cell growth were established. Our results contribute to real-time non-invasive methodologies for monitoring oxidative processes in biomedicine and biotechnology.

Genome-wide circulating tumor DNA monitoring for bladder cancer treatment management and organ preservation.

e16527 Background: Bladder cancer (BC) is the 9th most commonly diagnosed cancer worldwide and each year responsible for 165,000 deaths. Neoadjuvant combination chemotherapy, followed by radical cystectomy, is used for the management of localized muscle-invasive bladder cancer. One of the critical challenges in this therapeutic regimen is monitoring the tumor load to assess therapeutic efficacy – this is typically performed by assessing pathological downstaging in the cystectomy specimen. A high frequency of patients presents with T0N0 at cystectomy (no indication of residual disease), and consequently, it is vital to investigate organ preservation approaches to identify those patients who may qualify for bladder preservation. For precision oncology, we need to develop quantitative and non-invasive diagnostic methodologies to help the oncologist tailor the treatments to individual patients and monitor them for further clinical decision-making. Methods: Cell-free DNA (cfDNA) mutation detection has shown significant promise in its ability to monitor minimal residual disease and disease relapse by detection of cancer mutations in the peripheral blood. However, the combination of low tumor fraction and limited input material obtained from a typical plasma sample restricts the probability of detecting low metastatic burden in cfDNA through current deep targeted sequencing methods. Results: Here we present results from applying whole-genome sequencing (WGS) of cfDNA. We integrate a genome-wide mutation and copy number monitoring approach coupled with advanced signal processing and Artificial Intelligence (AI) for measuring the tumor load from low-input blood samples (̃1mL of plasma) with ultra-sensitive detection. The increased sensitivity allowed clinical detection of tumor fraction down to 8*10-5 and recurrence detection sensitivity achieving &gt; 65% at the first two months post-surgery. The WGS cfDNA approach is being evaluated on a patient cohort of more than 50 bladder cancer patients with longitudinal plasma sampling during neoadjuvant chemotherapy (response measure), pre-cystectomy (complete response measure), and post-surgery (relapse monitoring). Conclusions: The results indicate the clinical potential of genome-wide mutation integration as an ultra-sensitive, non-invasive diagnostic method for bladder cancer clinical management and bladder organ preservation.

Analytical Approach for the Study of Teotihuacan Mural Paintings from the Techinantitla Complex

Techinantitla building complex, in the Amanalco neighborhood of the ancient city of Teotihuacan, is famous for the iconography and quality of the mural paintings found in this site. A significant part of this heritage has been lost due to looting. In recent years, an interdisciplinary research project was developed to study the limited patrimony that was left. As part of this study, we first employed geophysical techniques to reconstruct the architectural pattern of the compound’s remaining walls, where other paintings may still be found. Then, we applied a non-invasive methodology to characterize a large set of fragments recovered in the 1980s and to gain information on their pigments and manufacturing techniques. This methodology included False Color Infrared Imaging, X-ray Fluorescence and Fiber-Optics Reflectance Spectroscopy, and led to the identification of hematite, calcite, malachite, azurite and an unidentified blue pigment. The results were compared with a previous study performed on a set of Techinantitla mural paintings looted in the 1960s. A broader comparison with contemporary mural paintings from other Teotihuacan complexes shows good agreement in the materials used. These results may suggest a standardization in the making of Teotihuacan mural painting during the Xolapan period (350 to 550 AD).

Can perioperative hemodilution be monitored with non-invasive measurement of blood hemoglobin?

BackgroundTrends in non-invasive measurements of blood hemoglobin (Hb) may be useful for identifying the need for transfusion in the perioperative period.MethodsCrystalloid fluid (5–20 mL/kg) was administered intravenously or by mouth to 30 volunteers and 33 surgical patients in five non-randomized clinical studies where Hb was measured on 915 occasions by non-invasive (Radical-7™) and invasive methodology. The hemodilution curves were compared by volume kinetic analysis and linear regression, with the slope and scattering of the data as key outcome measures.ResultsThe slope was 1.0, indicating unity between the two modes of measuring Hb when crystalloid fluid was infused in volunteers; however, only 40–45% of the variability in the non-invasive Hb could be explained by the invasive Hb. Patients undergoing major surgery, who showed the most pronounced hemodilution (median 24 g/L); non-invasive Hb explained 72% of the variability but indicated only half the magnitude of the invasive Hb changes (slope 0.48, P < 0.001 versus the volunteers). Simulations based on volume kinetic parameters from the volunteers showed 25% less plasma volume expansion after infusion when based on non-invasive as compared to invasive Hb, while no difference was found during infusion.ConclusionsIn volunteers the non-invasive Hb had good accuracy (low bias) but poor precision. In surgical patients the non-invasive Hb had good precision but systematically underestimated the hemodilution. Despite severe limitations, the non-invasive technology can be used to follow Hb trends during surgery if supported by occasional invasive measurements to assure acceptable quality of the hemodilution curve.Trial registrationsControlledTrials.gov NCT01195025, NCT01062776, NCT01458678, NCT03848507, and NCT01360333 on September 3, 2010, February 4, 2010, October 25, 2011, February 20, 2019, and May 25, 2011, respectively.

252 Non-Invasive Quantification of Human Brain Lactate Concentrations Across Sleep-Wake Cycles

Abstract Introduction Cellular mechanisms underlying changes in small animal brain lactate concentrations have been investigated for more than 70 years and report sharp reductions in lactate (12-35%) during sleep or anesthesia relative to wakefulness. The goal of this study was to investigate alterations in human cerebral lactate concentrations across sleep-wake cycles. Toward this goal, we developed a novel non-invasive methodology, quantified changes in human cerebral lactate during sleep stages, and investigated potential mechanisms associated with changes in lactate. Methods Nine subjects (four females, five males; 21-27 y-o, mean age 24.2 ±2) were sleep deprived overnight, and underwent (5:45~11:00 am) experiments combining simultaneous MR-spectroscopy (MRS) and polysomnography (PSG) in a 3 T MR instrument using a 64-channel head/neck coil. A single voxel MRS (1H-MRS) acquired signals from a volume of interest (12~24 cm3) for every 7.5-s for 88~180-min. Lactate signal intensity was determined from each 7.5-s spectrum, normalized to corresponding water signal, and averaged over 30-s for each PSG epochs. Artifact corrected PSG data were scored for each 30-s epoch using the standard criteria and classified into one of four stages: W, N1, N2 and N3. Group mean lactate levels were quantified using LCModel. Three subjects returned for lactate diffusivity measurements using diffusion-sensitized PRESS MRS sequence. Results Compared to W, group mean lactate levels within each sleep stage showed a reduction of [4.9 ± 4.9] % in N1, [10.4 ± 5.2] % in N2, and [24.0 ± 5.8] % in N3. We observed a significant decrease in lactate apparent diffusion coefficient (ADC) accompanied by reduced brain lactate in sleep compared to wake (P&amp;lt;0.002). There were no differences in ADC values between wake and sleep for H2O, NAA, tCr, or Cho. Conclusion This is the first in-vivo report of alterations in human brain lactate concentrations across sleep-wake cycles. Observed decline in lactate levels during sleep compared to wakefulness is consistent with, and extends results from invasive small animal brain studies first reported more than 70 years ago, and support the notion of altered lactate metabolism and/or increased glymphatic activity in sleeping human brain. Support (if any) The Paul. G. Allen Family Foundation funded the study.

Assessment of Steroid Hormones in Both Saliva and Blood During a Phase 2 Clinical Trial for the Use of Tildacerfont in Adults With Classic Congenital Adrenal Hyperplasia

Background: Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency is an autosomal recessive disorder characterized by an inability to synthesize endogenous cortisol resulting in overproduction of 17-hydroxyprogesterone (17-OHP) and androgens such as androstenedione (A4). Serum-based methods to measure steroid hormones typically require clinic visits and are not amenable to frequent, serial measurements when patient daily glucocorticoid doses are adjusted. Reliable, non-invasive new methodologies that can easily be adopted in a clinical trial setting are needed to complement existing measures of disease control. Here we report on the correlation between salivary and serum steroid hormone data from a Phase 2 proof-of-concept study for tildacerfont, a second generation corticotropin-releasing factor-1 (CRF1) receptor antagonist, which previously showed an ability to reduce excess adrenocorticotropic hormone (ACTH), 17-OHP, and A4 concentrations. Methods: Subjects with CAH under evaluation for tildacerfont underwent concurrent salivary and serum concentration measurements of androstenedione (A4), 17-hydroxyprogesterone (17-OHP) and testosterone (T) at approximately 8 am every 2 weeks for up to 6 weeks. Both serum and saliva samples were measured using liquid chromatography-tandem mass spectrometry. Results: 25 subjects (16 females) with a median age of 31 years (range 19-67) participated; median body mass index (BMI) was 27.8 kg/m2 (22-62 kg/m2). The number of matched samples across time points were: n=106 (17-OHP), n=106 (A4) and n=98 (T). 17-OHP and A4 were well correlated between serum and salivary assessments, r=0.80 (p<0.001) and r=0.80 (p<0.001), respectively, using kendall rank tests. T measurements had a lower correlation, r=0.67 (p<0.001). Over the duration of treatment, correlations between saliva and serum for 17-OHP and A4 were generally stable with only small differences across sex and clinic visit. Correlations between saliva and serum for T showed a moderate degree variability in women and a high degree of variability in men across visits. While correlations were high for A4 and 17-OHP, the magnitude of change, as a percentage of baseline across time points, was less pronounced with saliva as compared to serum. Conclusions: The ability to employ salivary steroid hormone measurements in CAH patients was demonstrated in a clinical trial setting. These data show that the measurement of the hormones 17-OHP and A4 in saliva may offer a promising, non-invasive approach to more frequently assessing response to therapy in patients with CAH.

Ultra high-frequency ultrasound with seventy-MHz transducer in hair disorders: Development of a novel noninvasive diagnostic methodology

BackgroundUltra high-frequency ultrasound (uHFUS) is a recently developed diagnostic technology. Despite its potential usefulness, no study has assessed its advantage in diagnosis and evaluation of hair disorders in comparison with other diagnostic methods. ObjectivesTo assess the practicability of uHFUS in diagnosing hair disorders and propose a diagnostic methodology. MethodsUltrasonographic images of scalp and forehead from patients with hair disorders (n = 103) and healthy controls (n = 40) were obtained by uHFUS and analyzed by both descriptive and numerical parameters. Furthermore, the data were compared with trichoscopic and histopathological findings. ResultsThe pattern of inflammation and fibrosis, hair cycle abnormality, and the findings in subcutis were detected by uHFUS. Significant differences were noted in the numerical parameters associated with the number of hair shafts and follicles, hair diameters and their diversity, and dermal echogenicity in both cicatricial and non-cicatricial hair disorders. Findings in uHFUS were associated with those observed in trichoscopy and scalp biopsy but uHFUS was able to detect pathological findings associated with hair cycle, inflammation, fibrosis, and subcutaneous abnormalities, which are hardly assessable by trichoscopy. ConclusionThe findings of this study highlighted usefulness of uHFUS in diagnosing hair disorders, while overcoming the weaknesses and limitations of other diagnostic tools.