A methodological primer of extracellular vesicles isolation and characterization via different techniques

Biology Methods and Protocols, Год журнала: 2024, Номер 9(1)

Опубликована: Янв. 1, 2024

Abstract We present four different protocols of varying complexity for the isolation cell culture-derived extracellular vesicles (EVs)/exosome-enriched fractions with objective providing researchers easily conducted methods that can be adapted many uses in various laboratory settings and locations. These are primarily based on polymer precipitation, filtration and/or ultracentrifugation, as well size-exclusion chromatography (SEC) include: (i) polyethylene glycol sodium chloride supplementation conditioned medium followed by low-speed centrifugation; (ii) ultracentrifugation medium; (iii) media through a 100-kDa exclusion filter; (iv) using standard commercial kit. techniques further purification sucrose density gradient centrifugation, or SEC if needed equipment is available. HEK293 SH-SY5Y cultures were used to generate containing exosomes. This was then depleted cells debris, filtered 0.2-µM filter, supplemented protease RNAse inhibitors prior exosomal isolation. The purified EVs immediately stably stored at 4°C (up week imaging intact EVS downstream) −80°C extended periods biochemical study. Our aim not compare these methodologies but them descriptors so choose “best method” their work under individual conditions.

Язык: Английский

---

Salivary Biomarkers for Parkinson’s Disease: A Systematic Review with Meta-Analysis

Cells, Год журнала: 2024, Номер 13(4), С. 340 - 340

Опубликована: Фев. 14, 2024

Parkinson’s Disease (PD) is a common neurodegenerative disease which manifests with motor features, such as bradykinesia, resting tremor, rigidity, and postural instability. Using the non-invasive technique of saliva collection, we designed systematic review to answer question “Are salivary biomarkers reliable for diagnosis Disease?”. Following inclusion exclusion criteria, 30 studies were included in this (according PRISMA statement guidelines). Mostly proteins reported potential saliva. Based on meta-analysis, PD patients, levels total alpha-synuclein significantly decreased, those oligomeric increased. Also, according pooled AUC, heme oxygenase-1 demonstrated significant predictive value saliva-based diagnosis. In conclusion, some biomarkers, especially alpha-synuclein, can be altered could reliably useful early differentiating other synucleopathies.

Язык: Английский

---

Comparative Analysis of Data‐Driven Rescoring Platforms for Improved Peptide Identification in HeLa Digest Samples

PROTEOMICS, Год журнала: 2025, Номер unknown

Опубликована: Фев. 2, 2025

ABSTRACT Mass spectrometry is a critical tool to understand complex changes in biological processes. Despite significant advances search engine technology, many spectra remain unassigned. This research evaluates the performance of three rescoring platforms, Oktoberfest, MS 2 Rescore, and inSPIRE, using MaxQuant output. The results indicated substantial increase identifications at peptide level (40%–53%) PSM (64%–67%). However, some peptides were lost due limitations processing posttranslational modifications (PTMs)—with up 75% exhibiting PTMs. Each platform displayed distinct strengths weaknesses. For instance, inSPIRE performed best terms unique peptides, while Rescore better for PSMs higher FDR values. Differences stemmed from different sources: original feature selection, type ion series predicted, retention time predictor, PTMs compatibility. Overall, showed superior ability harness results. Taken all together, platforms clearly outperformed results; however, they demanded additional computation (up 77%) manual adjustments. findings here underline necessity integrating into current proteomics pipelines but also address challenges their implementation optimization. Future integrated may help enhance adoption.

Язык: Английский

---

Metabolomic and Proteomic Profiling of Serum-Derived Extracellular Vesicles from Early-Stage Amyotrophic Lateral Sclerosis Patients

Journal of Molecular Neuroscience, Год журнала: 2025, Номер 75(1)

Опубликована: Фев. 15, 2025

Язык: Английский

---

Systematic review and meta-analysis on microRNAs in amyotrophic lateral sclerosis

Brain Research Bulletin, Год журнала: 2023, Номер 194, С. 82 - 89

Опубликована: Янв. 18, 2023

MicroRNAs (miRNAs) exhibit a crucial role in the pathogenesis and progress of neurodegenerative disorders. Recent studies have shown abnormal levels miRNA expression patients with amyotrophic lateral sclerosis (ALS). Clinical data also confirmed that miRNAs these are inconsistent across studies. A comprehensive systematic review meta-analysis current can help recognize important roles during ALS development. Therefore, we initially aimed to perform literature on muscle or serum healthy individuals. Subsequently, quantitatively summarized clinical individuals using meta-analytical technique. 11 comprising 281 244 control (HC) controls were identified from PubMed Web Science for meta-analysis. revealed closely associated occurrence disease. The most relevant either increased decreased. random-effects indicated miR-206, miR-133b, miR-338–3p significantly elevated than HC subjects. By contrast, there was no significant differences miR-133a between Collectively, our outcomes demonstrated ALS. We speculated miR-133b potential promising biomarkers

Язык: Английский

---

Discovery of Biomarkers for Amyotrophic Lateral Sclerosis from Human Cerebrospinal Fluid Using Mass-Spectrometry-Based Proteomics

Biomedicines, Год журнала: 2023, Номер 11(5), С. 1250 - 1250

Опубликована: Апрель 23, 2023

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by the loss of upper and lower motor neurons, which eventually may lead to death. Critical mission developing effective therapies for ALS discovery biomarkers that can illuminate mechanisms neurodegeneration have diagnostic, prognostic, or pharmacodynamic value. Here, we merged unbiased discovery-based approaches targeted quantitative comparative analyses identify proteins are altered in cerebrospinal fluid (CSF) from patients with ALS. Mass spectrometry (MS)-based proteomic employing tandem mass tag (TMT) quantification methods 40 CSF samples comprising 20 healthy control (HC) individuals identified 53 differential between two groups after fractionation. Notably, these included both previously ones, validating our approach, novel ones potential expanding biomarker repertoire. The were subsequently examined using parallel reaction monitoring (PRM) MS on 61 unfractionated 30 31 HC individuals. Fifteen (APOB, APP, CAMK2A, CHI3L1, CHIT1, CLSTN3, ERAP2, FSTL4, GPNMB, JCHAIN, L1CAM, NPTX2, SERPINA1, SERPINA3, UCHL1) showed significant differences control. Taken together, this study multiple ALS, providing foundation new

Язык: Английский

---

A Review of the Common Neurodegenerative Disorders: Current Therapeutic Approaches and the Potential Role of Bioactive Peptides

Current Protein and Peptide Science, Год журнала: 2024, Номер 25(7), С. 507 - 526

Опубликована: Апрель 2, 2024

Abstract:: Neurodegenerative disorders, which include Alzheimer's disease (AD), Parkinson's (PD), Huntington's (HD), and amyotrophic lateral sclerosis (ALS), represent a significant growing global health challenge. Current therapies predominantly focus on symptom management rather than altering progression. In this review, we discuss the major therapeutic strategies in practice for these highlighting their limitations. For AD, mainstay treatments are cholinesterase inhibitors N-methyl-D-aspartate (NMDA) receptor antagonists. PD, dopamine replacement therapies, including levodopa, commonly used. HD is managed primarily with symptomatic treatments, reusable extends survival ALS. However, none of halts or substantially slows neurodegenerative process. contrast, review highlights emerging research into bioactive peptides as potential agents. These naturally occurring synthetically designed molecules can interact specific cellular targets, potentially modulating processes. Preclinical studies suggest that may mitigate oxidative stress, inflammation, protein misfolding, common pathological features diseases. Clinical trials using neurodegeneration limited but show promising initial results. instance, hemiacetal, γ-secretase inhibitor peptide, has shown AD by reducing amyloid-beta production, though its development was discontinued due to side effects. Despite advancements, many challenges remain, identifying optimal peptides, confirming mechanisms action, overcoming obstacles related delivery brain. Future should prioritize discovery novel improve our understanding pharmacokinetics pharmacodynamics. Ultimately, approach lead more effective moving beyond modify course devastating

Язык: Английский

---

Prospect of extracellular vesicles in tumor immunotherapy

Frontiers in Immunology, Год журнала: 2025, Номер 16

Опубликована: Фев. 26, 2025

Extracellular vesicles (EVs), as cell-derived small vesicles, facilitate intercellular communication within the tumor microenvironment (TME) by transporting biomolecules. EVs from different sources have varied contents, demonstrating differentiated functions that can either promote or inhibit cancer progression. Thus, regulating formation, secretion, and intake of becomes a new strategy for intervention. Advancements in EV isolation techniques spurred interest EV-based therapies, particularly immunotherapy. This review explores multifaceted various immunotherapy, highlighting their potential vaccines adoptive cell therapy. Furthermore, we explore nanoparticle delivery systems Finally, discuss current state clinical settings future directions, aiming to provide crucial information advance development application treatment.

Язык: Английский

---

The Need for Biomarkers in the ALS–FTD Spectrum: A Clinical Point of View on the Role of Proteomics

Proteomes, Год журнала: 2023, Номер 11(1), С. 1 - 1

Опубликована: Янв. 9, 2023

Frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS) are severely debilitating progressive neurodegenerative disorders. A distinctive pathological feature of several diseases, including ALS FTD, is the deposition aberrant protein inclusions in neuronal cells, which leads to cellular dysfunction damage loss. Despite this, date, biological process behind developing these must be better clarified, making development disease-modifying treatment impossible until this done. Proteomics a powerful tool characterize expression, structure, functions, interactions, modifications proteins tissue fluid, plasma, serum, cerebrospinal fluid. This protein-profiling characterization aims identify disease-specific alteration or specific pathology-based mechanisms may used as markers conditions. Our narrative review highlight need for biomarkers potential use proteomics clinical practice ALS-FTD spectrum disorders, considering emerging rationale new drug development. Certainly, data will emerge near future regard support clinicians personalized medicine.

Язык: Английский

---

A Comprehensive Review of the Role of Biomarkers in Early Diagnosis of Parkinson’s Disease

Cureus, Год журнала: 2024, Номер unknown

Опубликована: Фев. 16, 2024

Parkinson’s disease (PD) is a complex neurological, degenerative clinical condition depicted by the advancing loss of dopaminergic neurons in substantia nigra pars compacta, which manifests itself as myriad sensorimotor and non-motor signs patients. The occurs due to reduced levels neurotransmitter dopamine brain, primarily associated with functional characteristics regarding mobility cognition. basal ganglion mainly involved generation cognitive functions therefore most significantly area PD. Since classical diagnosis assessment PD depends majorly on appearance motor characteristics, only arise when ~60-80% neuronal cell death has already occurred, it imperative we focus identifying biomarkers that can help us assess diagnose earlier stages progression, thus providing better prognosis for This review article will different are currently available use, divided under headings clinical, biological, imaging, genetic biomarkers, their specificity sensitivity toward an early Parkinson's patients future preclinical diagnostics using molecular biomarkers. affects over 1% population worldwide ranks second Alzheimer's context its incidence consequent socioeconomic burden. While recent breakthroughs have dramatically improved patients' odds survival prognosis, still remains symptomatic diagnostic tool. It research requires creating more advanced approaches diagnosing early, involving diagnostics, neuroimaging technology, biology collaborations provide highest degree care quality life patient deserves.

Язык: Английский

---