Journal of biological methods最新文献

Background: Cardiovascular diseases remain the leading cause of mortality globally, driven largely by modifiable risk factors, such as hyperlipidemia, hypertension, diabetes, and oxidative stress. As conventional pharmaceutical interventions often carry adverse effects and economic burdens, there is growing interest in alternative therapies.

Objective: This narrative review explores vinegar, a traditional remedy with historical and cross-cultural usage, as a potential adjunct in cardiovascular health management. Vinegar contains a rich array of bioactive compounds, including acetic acid, polyphenols, amino acids, melanoidins, and tetramethylpyrazine, which may exert antioxidant, anti-inflammatory, lipid-lowering, antihypertensive, and antithrombotic effects. Preclinical studies in animal models and limited human trials indicate promising outcomes in improving lipid profiles, blood pressure regulation, glycemic control, and endothelial function. Despite these benefits, there is a paucity of large-scale, high-quality human studies to validate vinegar's clinical efficacy and safety.

Conclusion: This review underscores the potential of vinegar as a functional food with cardioprotective properties and advocates for further rigorous research to establish standardized dosing, isolate active constituents, and assess long-term outcomes in human populations.

Background: The ability to test samples for the presence of specific tissues is useful for numerous forensic applications. More specifically, the identification of vital organ remains, such as the brain, in a crime scene or battlefield, can assist in determining the death of a missing person. In many cases, tissue samples are of insufficient quality or quantity for the application of histological methods, leaving forensic labs mostly restricted to immunological and catalytic assays designed to identify blood, semen, and saliva. Recent studies have suggested expression profiling-based methods for tissue and bodily fluid identification.

Objective: We present a methylation-based assay for the detection of brain tissue in forensic samples.

Methods: Genome-wide methylation data from 12 human tissues were analyzed to identify CpG sites uniquely methylated in brain tissue. Four candidate regions were selected based on high inter-tissue specificity and low intra-tissue variability. Targeted assays were developed using bisulfite conversion, polymerase chain reaction amplification, and next-generation sequencing, and validated based on reference tissues, mixtures, and environmentally degraded DNA samples.

Results: Four regions displayed consistent brain-specific methylation with >94% single-read accuracy and complete sample-level discrimination at ≥5% brain DNA. The assay retained diagnostic performance in mixed and degraded samples, demonstrating robustness under typical forensic conditions.

Conclusion: This study presents a sensitive and specific methylation-based assay for brain tissue identification. The approach enables reliable detection in degraded or composite materials and supports future integration of epigenetic biomarkers into forensic workflows for organ-source attribution.

Background: Prostate biopsy, while essential, often causes discomfort that can affect patient experience and adherence to follow-up procedures.

Objective: This study aimed to identify factors associated with pain during fusion prostate biopsy to optimize the experience of prostate cancer diagnosis and monitoring. The primary goal was to assess the relationship between pain during viscous lidocaine (lido) instillation and periprostatic nerve block with the overall pain experienced by patients undergoing prostate biopsy.

Methods: We queried our database for patients who underwent transrectal magnetic resonance imaging-ultrasound fusion prostate biopsy from March 2020 to July 2023 and had complete pain scores (1-10) recorded during lido instillation, periprostatic nerve block, biopsy, and overall.

Results: A total of 779 patients were included. The mean pain scores during lido instillation, periprostatic block, biopsy, and overall were 0.11, 2.8, 3.5, and 3.6, respectively. Multivariable analysis revealed that patients with a pain score during lido instillation of >2 (odds ratio [OR] = 10.28; p=0.027) patients with periprostatic block of >2 (OR = 7.49; p<0.001), black patients (OR = 2.838; p<0.001), and patients on active surveillance (OR = 1.648; p=0.003) were more likely to experience the upper quartile (UQ) of overall pain. Men with abnormal digital rectal examination (DRE) findings were less likely to develop the UQ of overall pain than men with normal DRE findings (OR: 0.586; p=0.004). This finding suggests that digital rectal examination during the initial clinic visit can help identify patients who may benefit from sedation during prostate biopsy, potentially improving patient comfort and procedural experience.

Conclusion: This finding suggests that digital rectal examination during the initial clinic visit can help identify patients who may benefit from sedation during prostate biopsy, potentially improving patient comfort and procedural experience.

Background: Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by deficits in social communication and the presence of restricted or repetitive behaviors. Although its underlying pathophysiological mechanisms remain unclear, growing evidence indicates that neuroinflammation plays a significant role, especially in children.

Objective: This study aims to explore neuroinflammatory pathways in children aged 12 and under, with a focus on potential therapeutic opportunities through drug repositioning.

Methods: We conducted a systematic computational analysis using data from 27 studies and bioinformatics resources such as DrugBank and PubChem, identifying over 8,000 potential drug candidates from the initial 29 treatments retrieved from the literature.

Results: Key compounds such as cannabidiol, fluoxetine, and risperidone were highlighted for their broad therapeutic potential. In addition, emerging treatments, including cell-based therapies and dietary interventions, were explored.

Conclusion: Our findings support drug repositioning as an effective strategy for developing new ASD treatments during critical developmental periods, emphasizing the need for further research to validate these pathways and the efficacy of innovative therapies.

Background: Chronic pain poses a significant challenge to the general population, especially in older adults. As a result, there is a growing emphasis on developing novel treatment options and expanding the use of emerging technologies.

Objective: This comprehensive review aims to guide practitioners in chronic pain management by extending the current application of vagus nerve stimulation (VNS). The review examined peer-reviewed studies, including multicenter cohort studies and clinical trials, published within the past 25 years. VNS has shown the most promising results in pain reduction for headache and migraine, with less relief in inflammatory and neuropathic pain as compared to current first-line treatment options.

Conclusion: Overall, this review seeks to compile and critically analyze the effect and efficacy of VNS on different forms of chronic pain using the most current studies and research applicable.

Background: Endometriosis represents a predominant gynecological disease that globally impacts 10% of women in their reproductive years, often leading to pelvic pain, infertility, and other complications.

Objective: This review intended to impart a comprehensive understanding of the pathophysiology, inherent genetic susceptibility, and the role of stem cells in the progression of endometriosis. It explores the diagnostic challenges posed by the diverse presentation of lesions and the involvement of genetic factors, including genes related to inflammation, immune response, steroidogenesis, neo-angiogenesis, and DNA repair. In addition, the role of adult stem cells, particularly from bone marrow and the endometrium is highlighted as an important aspect of disease progression. The review also examined how environmental factors, including early menarche, heavy menstrual flow, and Müllerian anomalies, contribute to endometriosis development. The impact on fertility is discussed concerning pelvic anatomical distortions, which affect egg release, sperm motility, and embryo transit. Furthermore, the review addressed complications, such as chronic pelvic pain, dysmenorrhea, dyspareunia, and potential obstetric issues, including an increased risk of certain cancers. Finally, it emphasized the need for improved diagnostic techniques and targeted therapies, focusing on early detection, innovative treatments, and fertility preservation.

Conclusion: Advancements in genomic and molecular research are crucial to understanding the genetic basis of endometriosis and ultimately enhance the quality of life for those affected.

Background: Protein A affinity chromatography represents the most extensively used technique for initial product capture in antibody purification. The ligands of commercial Protein A resins from different, or even the same, vendors may exhibit distinct binding specificity. For example, MabSelect SuRe LX and MabSelect PrismA bind the antibody's fragment crystallizable region and both the fragment crystallizable and variable heavy chain 3 (VH3) regions, respectively, while MabSelect VH3, a newly launched Protein A resin, binds the VH3 region exclusively.

Objective: This study aimed to compare the capabilities of three Protein A resins-namely, MabSelect SuRe LX, MabSelect PrismA, and MabSelect VH3-in removing byproducts associated with a variable domain of heavy chain-only antibody-based trispecific antibody (TsAb).

Methods: Clarified culture harvest containing the TsAb was processed separately using MabSelect SuRe LX, MabSelect PrismA, and MabSelect VH3 columns. For each column, byproduct separation was monitored by analyzing relevant elution fractions with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size-exclusion chromatography-high-performance liquid chromatography.

Results: MabSelect VH3 demonstrated superior byproduct removal performance compared with the other two Protein A resins.

Conclusion: MabSelect VH3 is particularly suitable for the purification of bispecific antibodies and TsAbs, where the product and byproducts contain different numbers of VH3 domains. For multispecific antibody purification, screening different affinity resins with distinct binding specificity is recommended, as this can help identify the most effective option for separation.

Background: Blood is central to immune defense, rendering accurate assessment of its immunoreactivity vital for medical and biotechnological applications.

Objective: This study presented a novel whole blood immunoreactivity assay (WBIA) designed to mimic natural physiological conditions, preserving essential cell-cell and cell-cytokine interactions for ex vivo immunological analysis.

Methods: Fresh whole blood (with or without heparin) was stimulated with lipopolysaccharide (LPS), concanavalin A (Con A), or both, activating innate and adaptive immunity. Cytokine levels were measured through enzyme-linked immunosorbent assay after incubation.

Results: Coagulation enhanced secretion of interleukin (IL)-2 and vascular endothelial growth factor (VEGF) in mitogen-stimulated samples. LPS induced tumor necrosis factor (TNF)-α, IL-6, and VEGF, while LPS + Con A co-stimulation produced the highest levels of interferon (IFN)-γ, IL-2, and IL-10. Peak cytokine concentrations were reached at 18 h, declining by 48-72 h. In 18 h LPS + Con A-stimulated serum blood samples from 30 healthy donors (19 women, 11 men, aged 30-55), cytokine levels (pg/mL, mean ± standard error of the mean) were as follows: IL-1β at (521 ± 62), IL-2 (24 ± 4), IL-6 (569 ± 43), IL-8 (277 ± 28), IL-10 (198 ± 35), IL-18 (293 ± 19), IFN-γ (227 ± 108), TNF-α (930 ± 126), and VEGF (655 ± 55).

Conclusion: The WBIA provides a reliable, physiologically relevant model for evaluating immune responses to stimuli. Its high fidelity to in vivo conditions makes it a valuable tool for testing immunomodulatory drugs and monitoring immune status in clinical settings.

Background: In vitro cell culture is essential for elucidating various signaling mechanisms and screening pharmacological agents to assess their safety and efficacy. However, cell proliferation and survival in culture can be significantly influenced by variations in the composition of the culture medium. For instance, variations in glucose and fetal bovine serum (FBS) concentrations can impact cell viability. Despite this, only a few studies have examined the impact of varied FBS and glucose concentrations in culture media on cell viability.

Objective: This study investigated the mechanisms and cellular effects of glucose and FBS deprivation in glioblastoma cell lines.

Methods: We systematically evaluated the impact of FBS and glucose deprivation on the proliferation and survival of rat C6 and human U-87 MG glioblastoma cell lines.

Results: Glucose deprivation (0 mg/dL) significantly reduced the viability of C6 cells and moderately lowered the viability of U-87 MG cells, with partial recovery upon glucose supplementation (100 mg/dL, 400 mg/dL). Notably, FBS deprivation (0%) exerted a more profound effect, inducing the accumulation of reactive oxygen species and extensive cell death in both cell lines. Restoration of FBS (1, 2, 4, 6, 8, and 10%) recovered cell viability and reduced oxidative stress. Furthermore, both glucose and FBS deprivation altered antioxidant enzyme expression and mitochondrial function. Glucose and FBS deprivation also differentially affected protein kinase B phosphorylation, suggesting metabolic stress-induced signaling modulation.

Conclusion: These findings highlight the differential responses of glioblastoma cells to glucose and FBS deprivation and underscore the importance of standardizing culture conditions, especially serum and glucose levels, when designing experiments involving glioblastoma cells.