Human Fluid Samples Research Articles

Development of a concentration method for simple and sensitive detection of SARS-CoV-2 in saliva using a magnetic nanoparticle kit

BackgroundWhen diagnosing viral infections in humans and animals, the presence of virus in a sample in trace amounts that are below the analytical sensitivity of the detection system may cause false negative results and inaccurate diagnosis. We previously reported the development of a simple virion concentration technique using 12ml large-volume samples that can dramatically improve diagnostic sensitivity by increasing analytical sensitivity by 100-fold over conventional methods. The present study was conducted to further improve the simplicity and versatility of this method. We constructed a simple and highly sensitive method for the detection of SARS-CoV-2 in human saliva after concentration using a magnetic nanoparticle conjugated with polyethylene glycol (PEG). ResultsPerformance of the method was evaluated by comparing a combination of automated nucleic acid extraction and RT-qPCR or triplex RT-LAM detection in a spiked sample of 20ml saliva collected from healthy humans. The method theoretically achieved 300-fold concentration of spiked SARS-CoV-2 in saliva, enabling 10- to 1000-fold higher analytical sensitivity for detection compared to conventional RNA extraction methods. ConclusionsThis newly developed method allows for easy and reliable concentration of the virion in less than 60min, improving the analytical sensitivity of the SARS-CoV-2 test. Further, the method allows for easy and reliable enrichment of the virus in less than 60min, improving the analytical sensitivity of the SARS-CoV-2 test. This method is easily used for highly sensitive virus detection from a variety of human oral fluid samples and may also be applied to rapid and labor-saving screening tests by pooling a large number of samples.

Characterisation of colloidal structures and their solubilising potential for BCS class II drugs in fasted state simulated intestinal fluid

A suite of fasted state simulated intestinal fluid (SIF), based on variability observed in a range of fasted state human intestinal fluid (HIF) samples was used to study the solubility of eight poorly soluble drugs (three acidic drugs (naproxen, indomethacin and phenytoin), two basic drugs (carvedilol and tadalafil) and three neutral drugs (felodipine, fenofibrate, griseofulvin)). Particle size of the colloidal structures formed in these SIF in the presence and absence of drugs was measured using dynamic light scattering and nanoparticle tracking analysis.Results indicate that drug solubility tends to increase with increasing total amphiphile concentration (TAC) in SIF with acidic drugs proving to be more soluble than basic or neutral drug in the media evaluated. Dynamic light scattering showed that as the amphiphile concentration increased, the hydrodynamic diameters of the structures decreased. The scattering distribution confirmed the polydispersity of the simulated intestinal fluids compared to the monodisperse distribution observed for FaSSIF v1). There was a large difference in the size of the structures found based on the composition of the SIF, for example, the diameter of the structures measured in felodipine in the minimum TAC media was measured to be 170 ± 5 nm which decreased to 5.1 ± 0.2 nm in the maximum TAC media point. The size measured of the colloidal structures of felodipine in the FaSSIF v1 was 86 ± 1 nm. However, there was no simple correlation between solubility and colloidal size.Nanoparticle tracking analysis was used for the first time to characterise colloidal structures within SIF and the results were compared to those obtained by dynamic light scattering. The particle size measured by dynamic light scattering was generally greater in media with a lower concentration of amphiphiles and smaller in media of a higher concentration of amphiphiles, compared to that of the data yielded by nanoparticle tracking analysis.This work shows that the colloidal structures formed vary depending on the composition of SIF which affects the solubility. Work is ongoing to determine the relationship between colloidal structure and solubility.

Comparative genomic analysis of an emerging Pseudomonadaceae member, Thiopseudomonas alkaliphila.

Thiopseudomonas alkaliphila, an organism recently classified within the Pseudomonadaceae family, has been detected in diverse sources such as human tissues, animal guts, industrial fermenters, and decomposition environments, suggesting a diverse ecological role. However, a large knowledge gap exists in how T. alkaliphila functions. In this comparative genomic analysis, adaptations indicative of habitat specificity among strains and genomic similarity to known opportunistic pathogens are revealed. Genomic investigation reveals a core metabolic utilization of multiple oxidative and non-oxidative catabolic pathways, suggesting adaptability to varied environments and carbon sources. The genomic repertoire of T. alkaliphila includes secondary metabolites, such as antimicrobials and siderophores, indicative of its involvement in microbial competition and resource acquisition. Additionally, the presence of transposases, prophages, plasmids, and Clustered Regularly Interspaced Short Palindromic Repeats-Cas systems in T. alkaliphila genomes suggests mechanisms for horizontal gene transfer and defense against viral predation. This comprehensive genomic analysis expands our understanding on the ecological functions, community interactions, and potential virulence of T. alkaliphila, while emphasizing its adaptability and diverse capabilities across environmental and host-associated ecosystems.IMPORTANCEAs the microbial world continues to be explored, new organisms will emerge with beneficial and/or pathogenetic impact. Thiopseudomonas alkaliphila is a species originally isolated from clinical human tissue and fluid samples but has not been attributed to disease. Since its classification, T. alkaliphila has been found in animal guts, animal waste, decomposing remains, and biogas fermentation reactors. This is the first study to provide an in-depth view of the metabolic potential of publicly available genomes belonging to this species through a comparative genomics and draft pangenome calculation approach. It was found that T. alkaliphila is metabolically versatile and likely adapts to diverse energy sources and environments, which may make it useful for bioremediation and in industrial settings. A range of virulence factors and antibiotic resistances were also detected, suggesting T. alkaliphila may operate as an undescribed opportunistic pathogen.

Dual-mode photothermal/chemiluminescence vertical flow assay for sensitive point-of-care detection of carcinoembryonic antigen using Cu2-xAgxS@liposome on a filter membrane

Conventional lateral flow assays based on colorimetry and fluorescence still have shortages in sensitivity and selectivity due to the severe background interference from complex human fluid sample matrices. In this work, Cu2-xAgxS nanocrystals with high photothermal conversion efficiency and good peroxidase-like activity were synthesized and applied in the construction of a dual-mode near-infrared-photothermal/chemiluminescence (CL) vertical flow assay of carcinoembryonic antigen (CEA). These two-mode principles showed nearly zero background and the synthesized Cu2-xAgxS exhibited a high photothermal conversion efficiency of 75.23%, enabling the luminol-H2O2 CL system to have over 4 min of chemiluminescence. By combining filter membrane enrichment, Cu2-xAgxS@liposome encapsulation amplification, and nanozyme catalysis, a dual-mode photothermal/CL portable assay was constructed for sensitive and accurate detection of CEA in serum, with linear ranges of 0.02–40 and 0.001–30 ng mL−1, and detection limits of 0.0023 and 0.00029 ng mL−1, respectively. Furthermore, a smartphone application and a 3D printing device were combined for point-of-care testing. This assay can be completed within 20 min, with simple operation and no need for large instruments. It exhibited good sensitivity, selectivity, and stability, and is expected to be used in early diagnosis and prevention of relevant diseases in resource-limited areas.

Identification of chronic non-atrophic gastritis and intestinal metaplasia stages in the Correa's cascade through machine learning analyses of SERS spectral signature of non-invasively-collected human gastric fluid samples

The progression of gastric cancer involves a complex multi-stage process, with gastroscopy and biopsy being the standard procedures for diagnosing gastric diseases. This study introduces an innovative non-invasive approach to differentiate gastric disease stage using gastric fluid samples through machine-learning-assisted surface-enhanced Raman spectroscopy (SERS). This method effectively identifies different stages of gastric lesions. The XGBoost algorithm demonstrates the highest accuracy of 96.88% and 91.67%, respectively, in distinguishing chronic non-atrophic gastritis from intestinal metaplasia and different subtypes of gastritis (mild, moderate, and severe). Through blinded testing validation, the model can achieve more than 80% accuracy. These findings offer new possibilities for rapid, cost-effective, and minimally invasive diagnosis of gastric diseases.

C. Halicacabum derived “turn-on-off” non-invasive molecularly imprinted polymer for selective profiling of 2-(3,4-dihydroxy phenyl) ethylamine in human fluid samples

A rationally designed “Turn on-off” molecularly imprinted fluorescent probe SCDs-MPs serves as a robust platform for the specific identification of aberrant dopamine (DA) levels. In this study, carbon dots (SCDs) were synthesized usingCardiospermum Halicacabum leaf extract as a carbon source via a step hydrothermal process. The synergistic integration of the synthesized fluorophore SCDs (QY–27.6 %) with molecularly imprinted elements via reverse micro-emulsion technique forms novel C. Halicacabum derived SCDs-MPs nanocomposite. The synthesized SCDs-MPs possess various inimitable properties such as hydrophilicity (WCA of 12.9°) suitability for detection in aqueous media, the narrow bandgap of 3.45 eV attributed to the electron-donating nature, and photostability. Fluorescence lifetime, from 6.38 ns to 6.17 ns after adding dopamine to SCDs-MPs, and zeta potential studies (–32.9 mV to –23.9 mV) validate the static fluorescence quenching mechanism in SCDs-MPs. Under various modifications and experimental conditions, SCDs-MPs consistently achieved a low dopamine detection limit of 6.4 pM within the range of 0–50 µM. Excellent recovery experiments in human fluid samples (98–102 %) confirm the fluorescent probe’s empirical reliability. The MTT assay confirms the superior biocompatibility of SCDs-MPs, with cell viability remaining at ± 24.03 % at a concentration of 200 µg/ml suggesting the safe usage for invitro administration. This opens new possibilities for utilizing C. Halicacabum derived SCDs-MPs material in biomedical contexts.

Tandem Mass Tag LC-MS/MS of Aqueous Humor From Individuals With Type 2 Diabetes Without Retinopathy Reveals Early Dysregulation of Synaptic Proteins.

An early neurodegenerative component of diabetic retinal disease (DRD) that precedes the vascular findings of clinically diagnosed diabetic retinopathy (DR) is increasingly being recognized. However, the relevant molecular mechanisms and biomarkers for early DRD are poorly defined. The purpose of this study was to uncover novel potential mediators of early diabetic retinal neuronal dysfunction through analysis of the aqueous fluid proteome in preclinical DR. Aqueous fluid was collected from subjects with type 2 diabetes mellitus (DM) but no clinical DR and from nondiabetic controls undergoing routine cataract surgery. Preoperative spectral-domain optical coherence tomography of the macula was obtained. Tandem mass tag LC-MS/MS was performed to identify proteins differentially present in diabetic and control aqueous fluid, and proteins with >50% change and P < 0.05 were considered significant. Selected results were validated with western blot of human aqueous fluid samples. We identified decreased levels of proteins implicated in neuronal synapse formation and increased levels of inflammatory proteins in the aqueous fluid from patients with type 2 DM but no DR compared with controls. Of the differentially present synaptic proteins that we identified and confirmed with western blot, the majority have not previously been linked with DRD. The proteomic profile of aqueous fluid from individuals with type 2 DM but no DR suggests that retinal neuronal dysfunction and inflammation represent very early events in the pathophysiology of DRD. These findings support the concept that diabetic retinal neurodegeneration precedes vascular pathology and reveal novel potential mediators and/or biomarkers warranting further investigation.

Immunoaffinity-free chromatographic purification of ovarian cancer biomarker CA125 (MUC16) from blood serum enables mass spectrometry characterization.

The enrichment of trace proteins from human fluid samples is of great importance in diverse clinical and industrial applications. In clinical diagnostics, such enrichment may enable detection of trace proteins that serve as biomarkers of disease. Affinity-based approaches, such as immunoaffinity pulldown, are widely used to enrich trace proteins, but this strategy relies on the availability and performance of antibodies that act on all proteoforms in an unbiased manner. Our prior work to characterize MUC16 (the mucin protein that carries the ovarian cancer biomarker CA125) by mass spectrometry successfully overcame the reliance on affinity-based enrichment and was used to enrich this biomarker from ascites of individual ovarian cancer patients, however, this strategy was not demonstrated on clinically relevant volumes of serum, a biofluid that is more accessible than ascites. The present work developed a non-affinity-based chromatographic method to enrich MUC16 from serum. The enriched MUC16 sample was further processed using a Midi Top 14 abundant protein depletion column. Peptides identified using bottom-up proteomics yielded 1-8% coverage of MUC16. Additionally, MUC16 was detected in samples containing less than the clinical cut-off level of CA125 (35 U mL-1), suggesting that this strategy of enrichment and bottom-up proteomics can enable analysis of CA125 from the serum of individuals with early-stage ovarian cancer and those whose tumors express CA125 (MUC16) at low levels.

Detection of gluten peptides in human duodenal fluids with immuno-liquid chromatography-mass spectrometry.

Gluten proteins are storage proteins in wheat that exhibit a certain resistance to gastrointestinal digestion. To explore solutions to cope with accidental ingestion of gluten in individuals suffering from gluten-related disorders, it is essential to monitor the fate of gluten peptides in biological samples, i.e., gastrointestinal juices, blood plasma or urine. In this work, we aimed at developing a mass spectrometry (MS)-based method for measuring gluten peptides in human duodenal fluids. Seven gluten peptides, including the well-documented 33-mer gluten peptide (LQLQPFPQPQLPYPQPQLPYPQPQLPYPQPQPF), were selected after a literature review and characterization of a gluten-containing product. Isotopically labelled peptides were used as references and a targeted liquid chromatography (LC) MS assay based on high resolution parallel reaction monitoring (PRM) was designed. Despite iterative and fine tuning of the LC-PRM-MS method, the low level of endogenous gluten peptides in human duodenal fluid samples precluded their direct detection. Thus, an initial immunoprecipitation (IP) step was included. Several antibodies were tested, and one proved reliable for the enrichment of the 33-mer gluten peptide as well as a few additional gluten peptides. Figures-of-merits of the immuno-LC-PRM-MS assay were assessed with a focus on quantification trueness and precision. We have developed an MS-based method for measuring the 33-mer gluten peptide in human duodenal fluids. Based on isotopic dilution, the method relies on the combination of IP and LC-PRM-MS analysis. Measurements were shown to be sensitive, quantitative, and reproducible.

The first LC-MS/MS stereoselective bioanalytical methods to quantitatevely detect 9R- and 9S-hexahydrocannabinols and their metabolites in human blood, oral fluid and urine

A sensitive LC-MS/MS method for the simultaneous quantification of the (9R)- and (9S)- hexahydrocannabinols (HHCs), and their metabolites, in human urine, oral fluid (OF) and blood samples were developed, validated and used to the biological samples of volunteers. The analytes were extracted from 100µL human samples. An isocratic elution mode with methanol was used for chromatographic separation of (9R)- and (9S)-HHC on an immobilized amylose tris(3-chloro-5-methylphenylcarbamate)-based chiral column Lux i-Amylose-3. The flow-rate of the mobile phase was 0.5mL/min. An isocratic elution mode of methanol and water (80/20, v/v) was used for chromatographic separation of metabolites of (9R)- and (9S)-HHC on a Lux AMP chiral column (with a proprietary chiral selector) at a flow rate of 0.5mL/min. MS/MS analysis was performed in positive ionization mode for HHC epimers, while in negative ionization mode was used for metabolites of HHCs. The calibration curves for HHCs and their metabolites in human samples ranged from 0.25- 240ngmL-1 and 1 - 100ngmL-1, respectively, with determination coefficients (r2) of ≥ 0.99. All analytes were stable at room temperature, 4 °C, in the autosampler (+10°C) and −20 °C for 24h, after three freeze/thaw cycles, and when stored at −20 °C up to one week after quality control (QC) sample preparation (concentration differences less than 20% with respect to time zero response), in blood, urine and OF.

Regulation of microRNA with food. Part 1. Food of plant origin

Тhe scientific review presents the mechanisms of microRNA regulation of biological processes in the human body with the help of food products of plant origin. To write the article, information was searched using Scopus, Web of Science, MedLine, PubMed, Google Scholar, EMBASE, Global Health, The Cochrane Library databases. The obtained information on dietary microRNAs is organized into the Dietary MicroRNA Databases (http://sbbi.unl.edu/dmd/), which is a repository for archiving and analyzing structural and functional microRNAs that enter the human body with food. It is stated that exogenous plant miRNAs entering the body with food are present in blood serum, tissues of humans and various animals, and regulate the expression of target genes. More than 50 miRNAs of plant origin have been identified in human blood serum using next-generation sequencing. The authors list some representatives from the multitude of dietary plant miRNAs. It is emphasized that several miRNAs show their functional activity both in plants and in mammals and easily overcome physiological barriers (miR-155, miR-168 and miR-854). Scientists believe that the ambiguous effect of plant products on the expression of human microRNA was shown by the results of a study conducted in vegetarians. Thus, with a comprehensive review using the latest information search databases, it was found that in modern scientific literature, the authors determine the horizontal transfer of numerous microRNA molecules from plants to the human body. Exogenous xenogenic xenomiRs are identified in most samples of human tissues and biological fluids. Entering the body together with food, they can significantly change the structure of the human transcriptome. Food products not only bring xenogeneic miRNAs, but also act as modulators of the endogenous miRNA generation by cells of the human body.

Granzyme B promotes matrix metalloproteinase-1 (MMP-1) release from gingival fibroblasts in a PAR1- and Erk1/2-dependent manner: A novel role in periodontal inflammation.

To gain insights into how proteases signal to connective tissues cells in the periodontium. The connective tissue degradation observed in periodontitis is largely due to matrix metalloproteinase (MMP) release by gingival fibroblasts. Granzyme B (GzmB) is a serine protease whose role in periodontitis is undefined. Human gingival crevicular fluid (GCF) samples were obtained from sites with periodontal disease and healthy control sites. GzmB was quantified in the GCF ([GzmB]GCF ) by ELISA. Gingival fibroblasts (GF) were cultured in the presence or absence of recombinant GzmB. Culture supernatants were analyzed by ELISA to quantify GzmB-induced release of interstitial collagenase (MMP-1). In some experiments, cells were pre-treated with the inhibitor PD98059 to block MEK/ERK signaling. The protease-activated receptor-1 (PAR-1) was blocked with ATAP-2 neutralizing antibody prior to GzmB stimulation. Systemic MMP-1 levels were measured in plasma from wild-type (WT) and granzyme-B-knockout (GzmB-/- ) mice. The [GzmB]GCF in human samples was ~4-5 fold higher at sites of periodontal disease (gingivitis/periodontitis) compared to healthy control sites, suggesting an association between GzmB and localized matrix degradation. GzmB induced a ~4-5-fold increase in MMP-1 secretion by cultured fibroblasts. GzmB induced phosphorylation of Erk1/2, which was abrogated by PD98059. GzmB-induced upregulation of MMP-1 secretion was also reduced by PD98059. Blockade of PAR-1 function by ATAP-2 abrogated the increase in MMP-1 secretion by GF. Circulating MMP-1 was similar in WT and GzmB-/- mice, suggesting that GzmB's effects on MMP-1 release are not reflected systemically. These data point to a novel GzmB-driven signaling pathway in fibroblasts in which MMP-1 secretion is upregulated in a PAR1- and Erk1/2-dependent manner.

Covalent organic frameworks based hierarchical porous hybrid monolithic capillary: Synthesis, characterization, and applications in trace metals analysis

The preparation of hierarchical porous monolithic column with high covalent organic frameworks (COF) loading and micropores accessibility is challenging due to the easy aggregability and sedimentation of COFs. Herein, a novel strategy based on high internal phase emulsion (HIPE) polymerization was proposed for preparing COF hybrid capillary monolithic column with hierarchical porosity. COFs with different frameworks including imine COFs (COF-OMe, COF-F and COF-SH), triazine COF (CTF-1) and boron-based COF (COF-5) were selected to investigate the universality of the preparation strategy. The presence of COF in the monolithic capillary was confirmed by scanning electron microscope, X-ray diffraction and fourier transform infrared spectroscopy. Nitrogen adsorption/desorption experiments and thermogravimetric analysis showed that the prepared COF hybrid monolithic capillary exhibited high COF loading (e.g., 28.7% for COF-SH) and accessibility (e.g., 98.5% for COF-SH), mainly due to the thin walls of void-window structures originated from polymerization of HIPE. The successful preparation of water-stable COF-F, COF-OMe, COF-SH and CTF-1 hybrid monolithic columns demonstrated the proposed synthesis strategy is universal to water-stable COF without tedious optimization of dispersion system, effectively avoiding the sedimentation of COF in pre-polymerization solution. Then, the sulfhydryl-modified COF hybrid polymer (poly(COF-SH-HIPE)) monolithic column was evaluated for the extraction of heavy metal ions, and a method based on poly(COF-SH-HIPE) monolithic capillary microextraction on-line coupled with inductively coupled plasma mass spectrometry detection was developed for analysis of trace Cd, Hg and Pb in human fluid samples.

Micro- and nanoplastics current status: legislation, gaps, limitations and socio-economic prospects for future

The pollution caused by micro- (MP) and nanoplastics (NP) in the planet’s ecosystems has gained significant interest in recent years due to their environmental impact and effects on the health of living organisms. Given this, it is necessary to conduct a comprehensive analysis of the actions required to mitigate their impacts. This paper analyzes existing legislation across different countries and regions, including Europe, North America, China, Russia, India, Brazil, Mexico, and the global initiatives undertaken by the United Nations. Furthermore, it highlights the need for additional measures to mitigate the impact of MP/NP in future years, such as the development of technologies for the separation or degradation of these particles in water intended for human consumption and in wastewater treatment plant effluents, studying plastic particulate material in the air considering meteorological parameters, MP/NP detection protocols in human fluid samples, creating truly biodegradable polymers for use as bioplastics, and establishing institutions responsible for the management of plastic waste. The study also shows the current state of abundance (characterization and quantification) of MP/NP in different environmental matrices based on reports from recent years, and identifies key research opportunities and actions required to evaluate the risks and toxicity associated with MP/NP. Socio-economic aspects are considered, including the impact of MP/NP on different regions, by associating economic and human wellness parameters to plastic waste generation by using available data from 148 countries. As result of this analysis, both the most populated and developed countries contribute to MP/NP generation, however, they have different capacities to address this problem due to social circumstances. The solution to this problem requires efforts from authorities, industry, the scientific community, and the active participation of the population, then, resolving social, political, and economic issues between countries and regions of the world is necessary.

RNA Sequencing Analysis of CD4+ T Cells Exposed to Airway Fluid From Children With Pediatric Acute Respiratory Distress Syndrome.

To identify differentially expressed genes and networks using a novel transcriptomic reporter assay with donor CD4+ T cells exposed to the airway fluid of intubated children with mild versus severe PARDS. In vitro pilot study. Laboratory-based study using human airway fluid samples admitted to a 36-bed university-affiliated pediatric intensive care unit. Seven children with severe PARDS, nine children with mild PARDS, and four intubated children without lung injury as controls. None. We performed bulk RNA sequencing using a transcriptomic reporter assay of CD4+ T cells exposed to airway fluid from intubated children to discover gene networks differentiating severe from mild PARDS. We found that innate immunity pathways, type I (α and β), and type II (γ) interferon response and cytokine/chemokine signaling are downregulated in CD4+ T cells exposed to airway fluid from intubated children with severe PARDS compared with those with mild PARDS. We identified gene networks important to the PARDS airway immune response using bulk RNA sequencing from a novel CD4+ T-cell reporter assay that exposed CD4+ T cells to airway fluid from intubated children with severe and mild PARDS. These pathways will help drive mechanistic investigations into PARDS. Validation of our findings using this transcriptomic reporter assay strategy is needed.

NiO Nanoparticle-decorated graphene oxide nanosheets modified glassy carbon electrode for sensitive electrochemical detection of pethidine

Pethidine (PTD), a synthetic opioid prescribed frequently as a painkiller, was included in the WADA list of prohibited substances in sports. The purpose of this study is to investigate the electrochemical determination of PTD using a modified glassy carbon electrode decorated with NiO nanoparticles and decorated graphene oxide nanosheets (NiO-GO/GCE). NiO-GO nanocomposite was created using a straightforward chemical reduction technique. SEM, TEM, and XRD analysis of the structure and morphology pointed to the NiO-GO nanocomposite's effective production. Results showed that NiO-GO/GCE responded to PTD determination in a sensitive, focused, and stable manner with a relatively low detection limit value (3.8 ng/mL) and a wide linear range (0–500 μg/mL) compared to other PTD sensors. Results on the appropriateness and validation of the NiO-GO/GCE for determining PTD content in clinical and human fluid samples illustrated good recoveries between 90.00 % and 99.66 % with a relative standard deviation less than 5.41 %, which were acceptable and corroborated the appropriateness and validation of the NiO-GO/GCE results for PTD sensor in urine samples from healthy athlete volunteers.

Molecular Screening for Urothelial Cancer: How Close We Are?

Early detection of urothelial cancer offers the potential for effective and successful treatment. Despite previous efforts, currently, there is not a well-validated, recommended screening program in any country. This integrative, literature-based review provides details on how recent molecular advances may further advance early tumor detection. The minimally invasive liquid biopsy is capable of identifying tumor material in human fluid samples from asymptomatic individuals. Circulating tumor biomarkers (cfDNA, exosomes, etc.) are very promising and are attracting the interest of numerous studies for the diagnosis of early-stage cancer. However, this approach definitely needs to be refined before clinical implementation. Nevertheless, despite the variety of current obstacles that require further research, the prospect of identifying urothelial carcinoma by a single urine or blood test seems truly intriguing.

Identification of Novel Biomarkers of Spinal Muscular Atrophy and Therapeutic Response by Proteomic and Metabolomic Profiling of Human Biological Fluid Samples.

Spinal muscular atrophy (SMA) is a neuromuscular disease resulting from mutations or deletions in SMN1 that lead to progressive death of alpha motor neurons, ultimately leading to severe muscle weakness and atrophy, as well as premature death in the absence of treatment. Recent approval of SMN-increasing medications as SMA therapy has altered the natural course of the disease. Thus, accurate biomarkers are needed to predict SMA severity, prognosis, drug response, and overall treatment efficacy. This article reviews novel non-targeted omics strategies that could become useful clinical tools for patients with SMA. Proteomics and metabolomics can provide insights into molecular events underlying disease progression and treatment response. High-throughput omics data have shown that untreated SMA patients have different profiles than controls. In addition, patients who clinically improved after treatment have a different profile than those who did not. These results provide a glimpse on potential markers that could assist in identifying therapy responders, in tracing the course of the disease, and in predicting its outcome. These studies have been restricted by the limited number of patients, but the approaches are feasible and can unravel severity-specific neuro-proteomic and metabolic SMA signatures.

Innovative, simple, and green: A sample preparation method based on 3D printed polymers

The present study evaluates the capability of fifteen 3D printed thermoplastic polymers as novel stationary phases for the extraction of forty-three physicochemically diverse analytes from fortified human oral fluid samples. Prototype extraction devices were prepared in 96-well plate-compatible format using fused deposition modeling 3D printer. The sample preparation was performed with 5-step protocol utilizing 96-well plates and semiautomated benchtop shaker. All resulting extracts were analyzed via high-performance liquid chromatography (operated in reversed-phase gradient elution mode) and tandem mass spectrometry (with electrospray ionization and triple quadrupole mass spectrometer). Exceptionally favorable results were observed for three polymer types: polyamide 6 (reinforced with 15% carbon fiber), LAYFOMM-60 (polyurethane with water-soluble polyvinyl alcohol), and S-FLEX 90A (thermoplastic polyurethane). Furthermore, this study also introduces an automated and repeatable 3D printing method for the fast fabrication of high-throughput, and highly selective sample preparation devices, most of which are ready-to-use without any additional processing or chemical functionalization. As such, the proposed printing method represents a significant step towards the introduction of novel polymeric stationary phases for analytical sample preparation, thus providing laboratory personnel with a method that is safer and more convenient, while minimizing negative environmental impacts.

Two-dimensional sulfur-doped carbon stacked sheets electrode design for potential adrenaline screening in human fluids and pharmaceutical drugs

An active electrochemical sensor for screening adrenaline (AD) in human fluid samples and pharmaceutical drugs is needed in clinical investigations and drug quality. The synthesis of sulfur-doped carbon microspheres (S-CMS) and irregular-edge carbon stacked sheet (S-CSS) structure was controlled based on the green synthesis of biomasses (i.e., lactose). The S-CSS intrinsic features of rough surface texture and stacked sheets with irregular edges were illustrated using various characterization techniques. The S-CSS was constructed with irregular edges around the stacked round sheets with a size range of 70–100 nm and a rough outer surface with ridged ends. The sulfur doping of S-CSS associated with a nanoporous network, and the high surface area ascertained a highly active transducing element for sensitive and selective electrode surface design. The designed electrode functionality and composition provide high electron/molecular diffusion and enhanced the electrode affinity and sensitivity. Therefore, the S-CSS-based sensor exhibited a fast charge transport surface, highly efficient electrocatalytic properties, and functionalized surfaces with numerous active centers for binding/sensing AD molecules. The S-CSS-based sensor exhibited a wide detection range (0.05–1.45 µM) and detection limit of 0.005 µM. AD-concentrations in blood samples and in pharmaceutical products have been tested using our designed electrode of S-CSS and obtained high selectivity and recovery of 102%. Thus, the S-CSS can be used to monitor AD in pharmaceutical drugs and human blood serum with fast response and using highly economic materials at extremely low concentrations.