Saliva diagnostics have become increasingly popular due to their non-invasive nature and patient-friendly collection process. Various collection methods are available, yet these are not always well standardized for either quantitative or qualitative analysis. In line, the objective of this study was to evaluate if measured levels of various biomarkers in the saliva of healthy individuals were affected by three distinct saliva collection methods: 1) unstimulated saliva, 2) chew stimulated saliva, and 3) oral rinse. Saliva samples from 30 healthy individuals were obtained by the three collection methods. Then, the levels of various salivary biomarkers such as proteins and ions were determined. It was found that levels of various biomarkers obtained from unstimulated saliva were comparable to those in chew stimulated saliva. The levels of potassium, sodium, and amylase activity differed significantly among the three collection methods. Levels of all biomarkers measured using the oral rinse method significantly differed from those obtained from unstimulated and chew-stimulated saliva. In conclusion, both unstimulated and chew-stimulated saliva provided comparable levels for a diverse group of biomarkers. However, the results obtained from the oral rinse method significantly differed from those of unstimulated and chew-stimulated saliva, due to the diluted nature of the saliva extract.
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Purpose: Exercise therapy with a focus on muscle strengthening has proven to be effective for the management of knee osteoarthritis (OA). Yet it is not known whether high-intensity resistance training (RT) is more effective in improving upper leg muscle strength and physical performance than low-intensity RT. Still, there is some controversy regarding the effectiveness of high-intensity RT and whether or not it is harmful, for instance by accelerating cartilage degeneration, osteophyte formation, or increasing synovitis. Any catabolic or anabolic response of musculoskeletal tissue to RT might first be visible on a biochemical level before changes in clinical symptoms are measurable. Serum biomarkers can objectively measure early biochemical changes and assess whether RT leads to a more anabolic or catabolic response. The aim of this study is to assess (i) whether high-intensity RT elicits a different response (e.g. catabolic) on systemic inflammation and musculoskeletal tissues in and surrounding the joint, including bone, cartilage, muscle, and synovial tissue compared to low-intensity RT; and (ii) whether there is an association between changes in serum levels of inflammatory and musculoskeletal tissue-derived biomarkers and improvements in clinical outcomes (performance-based tests and self-reported outcomes on pain and activity limitations).Methods: In a randomized controlled trial, 177 participants with knee OA conducted a high-intensity (70%-80% of the Repetition Maximum (1RM)) or low-intensity (40%-50% 1RM) RT program 3 times a week for 12 weeks. Measures of clinical outcomes and serum samples were collected at the start of RT (pre-intervention), after 3 months at the end of RT (post-intervention), and 6 months after RT (follow-up). As a reflection of systemic inflammation (CRP), synovitis (CRPM, C3M), bone turnover (OC, CTX-I), cartilage turnover (PRO-C2, C2M, huARGS), muscle turnover (PRO-C3, PRO-C6), and cell behaviour (col10neo) a total of eleven serum biomarkers were analysed. With the exception of CRP, which was determined with an immunoturbidimetric assay, ELISA assays were used to quantify serum levels of the other 10 serum biomarkers. The primary outcome measures are the changes in serum biomarker levels. Other outcome measures include upper leg muscle strength, performance-based tests, and self-reported outcomes on pain and activity limitations.Results: High-intensity RT resulted in greater improvements in muscle strength compared to low-intensity RT when measured by the estimated 1RM. No significant differences between groups were found for upper leg muscle strength (Nm/kg) when measured with an isokinetic dynamometer. Both groups showed similar improvements in pain and physical functioning. Although there is no difference between groups in clinical outcomes, except for the estimated 1RM, we expect that participants in the high-intensity RT group are more likely to have enhanced serum levels of catabolic biomarkers than participants in the low-intensity RT group. Since both the high-intensity RT group and low-intensity RT group improved over time, we expect that changes in serum biomarker levels are associated with overall improvements in clinical outcomes. Almost all participants had normal CRP values (<10 mg/L) at baseline. No significant differences between the intensity RT groups in CRP levels at baseline, at 3 months, and 6 months were found. In both groups, there was no evidence that RT influenced CRP serum levels.Conclusions: The work to date on CRP serum levels suggests that RT did not influence CRP levels. This result may be explained by the high percentage of participants with normal CRP levels (<10 mg/L). We are currently in the process of analyzing the remaining 10 neo-epitope biomarkers. We expect that our remaining 10 assays have the potential to measure changes in serum biomarker levels in response to RT. This will be the first study to investigate the effects of high-intensity versus low-intensity RT on musculoskeletal tissue turnover in individuals with knee OA. With this, we aim to determine whether high-intensity RT can improve upper leg muscle strength and physical performance without worsening systemic inflammation or causing adverse effects on musculoskeletal knee OA-related tissues.
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Objectives: Pulmonary hypertension is one of the leading causes of death in systemic sclerosis. Early detection and treatment of pulmonary hypertension in systemic sclerosis is crucial. Nailfold capillaroscopy microscopy, vascular autoantibodies AT1R and ETAR, and several candidate-biomarkers have the potential to serve as noninvasive tools to identify systemic sclerosis patients at risk for developing pulmonary hypertension. Here, we explore the classifying potential of nailfold capillaroscopy microscopy characteristics and serum levels of selected candidate-biomarkers in a sample of systemic sclerosis patients with and without different forms of pulmonary hypertension.Methods: A total of 81 consecutive systemic sclerosis patients were included, 40 with systemic sclerosis pulmonary hypertension and 41 with no pulmonary hypertension. In each group, quantitative and qualitative nailfold capillaroscopy microscopy characteristics, vascular autoantibodies AT1R and ETAR, and serum levels of 24 soluble serum factors were determined. For evaluation of the nailfold capillaroscopy microscopy characteristics, linear regression analysis accounting for age, sex, and diffusing capacity of the lungs for carbon monoxide percentage predicted was used. Autoantibodies and soluble serum factor levels were compared using two-sample t test with equal variances.Results: No statistically significant differences were observed in quantitative or qualitative nailfold capillaroscopy microscopy characteristics, or vascular autoantibody ETAR and AT1R titer between systemic sclerosis-pulmonary hypertension and systemic sclerosis-no pulmonary hypertension. In contrast, several serum levels of soluble factors differed between groups: Endostatin, sVCAM, and VEGFD were increased, and CXCL4, sVEGFR2, and PDGF-AB/BB were decreased in systemic sclerosis-pulmonary hypertension. Random forest classification identified Endostatin and CXCL4 as the most predictive classifiers to distinguish systemic sclerosispulmonary hypertension from systemic sclerosis-no pulmonary hypertension.Conclusion: This study shows the potential for several soluble serum factors to distinguish systemic sclerosis-pulmonary hypertension from systemic sclerosis-no pulmonary hypertension. We found no classifying potential for qualitative or quantitative nailfold capillaroscopy microscopy characteristics, or vascular autoantibodies.
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Cell-based production processes in bioreactors and fermenters need to be carefully monitored due to the complexity of the biological systems and the growth processes of the cells. Critical parameters are identified and monitored over time to guarantee product quality and consistency and to minimize over-processing and batch rejections. Sensors are already available for monitoring parameters such as temperature, glucose, pH, and CO2, but not yet for low-concentration substances like proteins and nucleic acids (DNA). An interesting critical parameter to monitor is host cell DNA (HCD), as it is considered an impurity in the final product (downstream process) and its concentration indicates the cell status (upstream process). The Molecular Biosensing group at the Eindhoven University of Technology and Helia Biomonitoring are developing a sensor for continuous biomarker monitoring, based on Biosensing by Particle Motion. With this consortium, we want to explore whether the sensor is suitable for the continuous measurement of HCD. Therefore, we need to set-up a joint laboratory infrastructure to develop HCD assays. Knowledge of how cells respond to environmental changes and how this is reflected in the DNA concentration profile in the cell medium needs to be explored. This KIEM study will enable us to set the first steps towards continuous HCD sensing from cell culture conditions controlling cell production processes. It eventually generates input for machine learning to be able to automate processes in bioreactors and fermenters e.g. for the production of biopharmaceuticals. The project entails collaboration with new partners and will set a strong basis for subsequent research projects leading to scientific and economic growth, and will also contribute to the human capital agenda.
Wetenschappelijk gezondheidsonderzoek is tot op heden met name uitgevoerd met mannen. Daarmee is de kennis rondom vrouwengezondheid achtergebleven. Er ligt een grote opgave om deze kennis te vergroten. Een van de uitdagingen is het versnellen van diagnose voor vrouwspecifieke aandoeningen. Een aandoening die aandacht vraagt voor versnelde diagnose is endometriose. Op dit moment wordt de diagnose endometriose gemiddeld pas na 8 jaar gesteld. Endometriose kan voor ondragelijke pijnklachten en onvruchtbaarheid zorgen wat grote impact heeft op het dagelijks functioneren en mentaal welzijn van vrouwen. Dit leidt ook tot verzuim. Het ontwikkelen van nieuwe diagnostische methodieken voor vrouwspecifieke aandoeningen is dus noodzakelijk om kwaliteit van leven en deelname aan de maatschappij te verbeteren. Er zijn aanwijzingen in de literatuur dat menstruatiebloed een bron van informatie kan zijn om de gezondheid van vrouwen te bepalen. Menstruatiebloed wordt momenteel vooral gezien als een afvalproduct. Hoewel dit mogelijk informatie bevat over de gezondheid van de baarmoeder, menstruatiegezondheid, hormonale gezondheid, of algehele gezondheid van vrouwen. Het is echter nog onvoldoende bekend wat de samenstelling van menstrueel bloed is. Dit onderzoek focust daarom op het onderzoeken van de samenstelling van menstruatiebloed. Deze kennis dient als verkenning voor vervolgonderzoek naar de mogelijkheid voor diagnose van gynaecologische aandoeningen via menstruatiebloed. Hiervoor is het eerst van belang om te achterhalen of er stabiele markers in menstruatiebloed aanwezig zijn die kunnen dienen als referentie voor diagnose. Daarom is de onderzoeksvraag van dit onderzoek: Welke stabiele factoren kunnen geïdentificeerd worden in menstruatiebloed, die als referentiemarker kunnen dienen voor diagnose? In dit onderzoek wordt hiervoor (1) een protocol ontwikkelt voor hygiënisch transport van menstruatiebloed van vrouw naar laboratorium én (2) geanalyseerd of er stabiele markers aanwezig zijn in menstruatiebloed. Beiden kunnen in vervolgonderzoek worden ingezet ter referentie aan aandoening-specifieke biomarkers.
Urineweginfecties behoren tot de meest voorkomende infecties waarvoor de huisarts wordt bezocht. Bij de diagnostiek van urineweginfecties wordt uitgegaan van een anamnese van klachten en vervolgens kunnen verschillende point-of-care testen worden ingezet. Deze testen zijn echter niet geschikt om de diagnose urineweginfectie te bevestigen, maar alleen (indien negatief) om de diagnose uit te sluiten. Daarnaast is het op basis van deze testen niet mogelijk om direct een gerichte therapie in te zetten. Er wordt regelmatig gestart met een antibioticumkuur zonder dat de verwekker bekend is, hetgeen kan leiden tot toename van de antibioticaresistentie. Er is grote behoefte aan nieuwe diagnostische benaderingen voor urineweginfecties. In het RAAK publiek project “Sneldiagnostiek van urineweginfecties in de eerstelijn” (RAAK.PUB04.050) is de inzet van single-cell MALDI-TOF massaspectrometrie onderzocht. Deze technologie blijkt in staat om zeer snel (in enkele minuten) de verwekker van een potentiële urineweginfectie te identificeren. De massaspectrometer levert unieke eiwitspectra van de bacteriecellen op. Regelmatig werd ook een additioneel laagmoleculair eiwitpatroon gevonden in de urine van patiënten met een urineweginfectie. Om die reden werd het onderzoek uitgebreid met de analyse van ruim 250 controlemonsters, urines van mensen zonder verdenking op een urineweginfectie. Uit dit onderzoek bleek dat de gevonden laag moleculaire peptiden in urine mogelijk gebruikt kunnen worden als biomarker voor een urineweginfectie. In het kader van het Top-up project willen we meer bekendheid geven aan de potentiële biomarker door middel van een wetenschappelijke publicatie. Daarnaast willen we nagaan of het mogelijk is om een pointof- care test te ontwikkelen op basis van de biomarker. Voor de klinische validatie van deze test wordt het netwerk gevormd en een onderzoeksplan voor een vervolgproject opgesteld.
