Experimental Biology and Medicine最新文献

The identification of epidermal growth factor receptor (EGFR) tyrosine kinase (TK) domain mutations in non-small cell lung cancer (NSCLC) patients is crucial for therapeutic decision-making and monitoring EGFR-tyrosine kinase inhibitor (TKI) resistance. Liquid biopsy has emerged as a promising alternative for patients ineligible for invasive tissue sampling. This study investigated the clinical utility of a novel chip-based digital PCR (dPCR) platform for detecting two important EGFR mutations - exon 19 deletions (19del) and threonine-methionine amino acid substitution at position 790 (T790M) - in serum samples, while exploring potential serum biomarkers for mutation prediction. The collection of 350 serum samples were conducted on patients diagnosed with NSCLC at Huashan Hospital between August 2023 and February 2024. Cell-free deoxyribonucleic acid (cfDNA) was extracted from serum and was analyzed for EGFR mutations using dPCR. The serum tumor marker levels were quantified. The dPCR assay demonstrated positive predictive values of 73.33% for 19del and 28.57% for T790M. Biomarker analysis revealed a carbohydrate antigen (CA) 199 cutoff of 11.75 U/mL (AUC = 0.707, 95% CI: 0.573-0.841, P = 0.005) for 19del detection, while progastrin-releasing peptide (ProGRP) showed a cutoff of 45.15 pg/mL (AUC = 0.628, 95% CI: 0.521-0.735, P = 0.028) for T790M identification. Variant rate exhibited significant positive correlations with biomarker concentrations: 19del variant rates significantly associated with CA125 levels (r = 0.624, P = 0.010), while T790M correlated with both carcinoembryonic antigen (CEA) (r = 0.531, P = 0.004) and ProGRP (r = 0.395, P = 0.041) in mutation-positive cohorts. These findings indicate that serum-based dPCR liquid biopsy demonstrates potential clinical utility as a supplementary approach to tissue biopsy for NSCLC genotyping. Notably, elevated serum tumor marker levels correlate with enhanced mutation detection rates in liquid biopsy, implying their potential supplementary value in prioritizing patients for molecular profiling.

The fatal neurodegenerative disease, amyotrophic lateral sclerosis (ALS), leads to the degeneration of motor neurons in the brain and spinal cord. Many different genetic variants are known to increase the risk of developing ALS, however much of the disease heritability is still to be identified. To identify novel genetic factors, we characterised SINE-VNTR-Alu (SVA) presence/absence variation in 4403 genomes from the New York Genome Center (NYGC) ALS consortium. SVAs are a type of retrotransposon able to mobilise in the human genome generating new insertions that can modulate gene expression and mRNA splicing and to date 33 insertions are known to cause a range of genetic diseases. In the NYGC ALS consortium sequence data 2831 non-reference genome SVAs were identified and 95% of these insertions were rare with an insertion allele frequency of less than 0.01. Association analysis of the common SVAs with ALS risk, age at onset and survival did not identify any SVAs that survived correction for multiple testing. However, there were three different rare SVA insertions in the ALS associated gene NEK1 identified in four different individuals with ALS. The frequency of these rare insertions in NEK1 was significantly higher in the individuals with ALS from the NYGC ALS consortium compared to the gnomAD SV non-neuro controls (p = 0.0002). This study was the first to characterise non-reference SVA presence/absence variation in a large cohort of ALS individuals identifying insertions as potential candidates involved in disease development for further investigation.

Clinically, reliably restoring meaningful peripheral sensory and motor nerve function across peripheral nerve gaps is limited. Thus, although autografts are the clinical "gold standard" repair technique for bridging nerve gaps, even under relatively good conditions, <50% of patients recover meaningful function. Due to this low recovery rate, many patients are not even provided repair surgery and, consequently, suffer permanent loss of function. This paper examines intrinsic and extrinsic changes associated with injured neurons and Schwann cells that reduce the extent of axon regeneration and recovery. It also examines how these changes can be reversed, leading to enhanced regeneration and recovery. It next examines the efficacy of platelet-rich plasma (PRP) in promoting axon regeneration and two novel techniques involving bridging nerve gaps with an autograft within a platelet-rich (PRP) collagen tube or only a PRP-filled collagen tube, which induce meaningful recovery under conditions where autografts alone are not effective. Finally, it looks at potential mechanisms by which platelet-released factors may enhance axon regeneration and recovery. This review shows that although there are many limitations to restoring meaningful function following peripheral nerve trauma, there are a number of ways these can be overcome. Presently, the most promising techniques involve using PRP.

Pharmacovigilance is essential for protecting patient health by monitoring and managing medication-related risks. Traditional methods like spontaneous reporting systems and clinical trials are valuable for identifying adverse drug events, but face delays in data access. Social media platforms, with their real-time data, offer a novel avenue for pharmacovigilance by providing a wealth of user-generated content on medication usage, adverse drug events, and public sentiment. However, the unstructured nature of social media content presents challenges in data analysis, including variability and potential biases. Advanced techniques like natural language processing and machine learning are increasingly being employed to extract meaningful information from social media data, aiding in early adverse drug event detection and real-time medication safety monitoring. Ensuring data reliability and addressing ethical considerations are crucial in this context. This review examines the existing literature on the use of social media data for drug safety analysis, highlighting the platforms involved, methodologies applied, and research questions explored. It also discusses the challenges, limitations, and future directions of this emerging field, emphasizing the need for ethical principles, transparency, and interdisciplinary collaboration to maximize the potential of social media in enhancing pharmacovigilance efforts.

The Acadian variant of Fanconi Syndrome (AVFS) is a rare genetic disorder characterized by renal deficiencies. AVFS is caused by a mutation to NDUFAF6 encoding a complex I assembly factor, and leading to metabolic alterations. We confirmed that fibroblasts derived from AVFS patients have lower complex I activity, mitochondrial membrane potential and cellular respiration. These mitochondrial defects were accompanied by higher levels of 8-hydroxy-2'deoxyguanosine, malondialdehyde and carbonyl, which are markers of oxidative damage to DNA, lipids and proteins, respectively. Thus, we hypothesized that the antioxidant N-Acetyl-L-cysteine (NAC) would reduce oxidative stress and mitochondrial defects in AVFS fibroblasts. Treatment with NAC during 5 days partially restored complex I activity, mitochondrial membrane potential and cellular respiration in AVFS fibroblasts. NAC also prevented oxidative damage in AVFS fibroblasts. This work shows for the first time that the physiopathology of AVFS includes high oxidative stress. It also reveals that NAC and other antioxidant-based strategies might represent an effective pharmacological treatment for AVFS.

Malaria remains a significant cause of childhood morbidity and mortality, with Plasmodium falciparum Histidine-Rich Protein 2 (PfHRP2)-based malaria rapid diagnostic tests (mRDTs) widely used in endemic regions where microscopy is sometimes not feasible. While these tests offer high sensitivity, persistent PfHRP2 antigenemia and gene deletions can cause false-positive and false-negative results, compromising their accuracy for malaria case management and surveillance. This study evaluated the diagnostic performance and antigen persistence of PfHRP2-mRDTs using data from a longitudinal birth cohort of 750 children followed monthly from birth to 36 months in a holoendemic region of Kenya. Malaria diagnosis was performed using both microscopy and mRDTs, with a total of 15,006 clinical events recorded from 573 children between 2017 and 2023. Data from an independent acute febrile cohort of 937 children (<5 years) followed for 14 days was analyzed to validate the findings. The mRDT showed a high sensitivity of 97.27% but a moderate specificity of 65.00% in acute febrile illness, indicating frequent false-positive results. The positive predictive value was low (35.10%), suggesting that confirmatory testing is needed, while the negative predictive value was high (98.89%), reinforcing the reliability of mRDTs in ruling out malaria. Persistent PfHRP2 antigenemia was observed, with a median antigen clearance time of 51.14 days, respectively. Higher initial parasite densities (>50,000/μL) were associated with a slower antigen decay rate (p = 0.001), highlighting the challenge of interpreting positive mRDT results after treatment. Validation using the acute febrile cohort showed that mRDT specificity exceeded 95% at initial diagnosis and follow-up. Overall, PfHRP2-based mRDTs remain valuable for frontline malaria diagnosis but are limited by antigen persistence, leading to false positives in follow-up testing. Where feasible, integration of confirmatory diagnostic methods, such as microscopy or molecular assays, could improve the performance of malaria case management and clinical decision making, particularly in high-transmission settings.

Exosomes are the smallest extracellular vesicles secreted from cells, carrying different cargos, including nucleic acids, proteins and others which transfer from cells to cells. The properties of exosomes depend on the donor cells. Hypoxia, referring to a sublethal and insufficient oxygen supply, reportedly influences exosome secretion of hypoxic cells. In the present study, we focused on the effects of hypoxia on exosomes obtained from CTX-TNA2 astrocyte cells exposed to different durations of hypoxia followed by normoxia as a model of hypoxic preconditioning. To evaluate the functions of exosomes, primary cultured cortical neurons were treated with hemin, a potent neurotoxin. Our sulforhodamine B assay showed that incubation of hemin (30 μM) consistently induced neuronal death. Co-incubation of exosomes from CTX-TNA2 cells subjected to 2 hr-hypoxia plus 6 hr-renormoxia (2H/6R exosomes), but not 12 hr-hypoxia plus 24 hr-renormoxia (12H/24R exosomes), attenuated hemin-induced cell death and reduction in growth associated protein 43 level (a biomarker of neurite outgrowth). Western blot assay demonstrated that 2H/6R exosomes attenuated hemin-induced elevations in inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) levels (two proinflammatory biomarkers) as well as heme oxygenase-1 (HO-1). In contrast, 12H/24R exosomes did not alter hemin-induced elevation in HO-1 but further augmented hemin-induced increases in iNOS and COX-2. Moreover, 2H/6R exosomes attenuated hemin-induced reduction in glutathione hydroperoxidase 4 (a biomarker of ferroptosis) and elevation in active caspase 3 (a biomarker of apoptosis) while 12H/24R exosomes did not effectively alter hemin-induced programed cell death. In conclusion, our study showed that 2H/6R exosomes possessed neuroprotective activities while 12H/24R exosomes had mild pro-inflammatory activities, suggesting that different hypoxic preconditionings influenced CTX-TNA2 cells which then secreted exosomes with differential biological activities. These findings highlight a double-edged role of hypoxia on exosome functions.

Mass cytometry enables high-throughput characterization of heterogeneous cell populations at single-cell resolution, using metal isotopes to capture cellular signals and avoiding the spectral overlap common in flow cytometry. Despite advancements, conventional data analysis often focuses on manual gating or clustering within specific samples, overlooking disparities across subjects or biological samples. To address this gap, we propose a novel framework that treats the cell-by-protein matrix as a high-dimensional distribution, using Quantized Optimal Transport (QOT) to quantify distances between samples based on their cellular protein expression profiles. This approach allows for a direct comparison of distributions without relying on predefined gating strategies, capturing subtle variations in the data. We validated our method through two experiments using real-world time-series Coronavirus Disease 2019 (COVID-19) cytometry data. First, we conducted a leave-one-out analysis to identify immunologically unstable proteins over time, revealing CD3 and CD45 as the proteins changing the most during the vaccine response. Second, we aimed to capture individual immune fingerprints over time by calculating pairwise Wasserstein distances between samples and applying hierarchical clustering. Using silhouette scores to evaluate clustering effectiveness, we identified optimal combinations of immunological markers that effectively grouped samples from the same participant across different time points. Our findings demonstrate that the QOT framework provides a robust and flexible tool for cohort-level analysis of mass cytometry data, enabling the identification of unstable immunological markers and capturing immune response heterogeneity among vaccinated cohorts.

Despite the highly effective impact of antiretroviral therapy (ART) in reducing mother-to-child transmission of human immunodeficiency virus (HIV), there are concerns of long-term impacts of ART on the health of the offspring. The implications of perinatal exposure to antiviral drugs on the gut bacterial population and metabolic function in the offspring is unclear but may influence health outcomes given the various reported effects of the microbiome in human health. This study aims to gain insight into the potential effect of in utero and lactational exposure to ART on gut microbiota populations and short-chain fatty acids (SCFAs) production in aged rat offspring. Pregnant rats were administered a combination of antiretroviral drugs (abacavir/dolutegravir/lamivudine) at two different dose levels during gestation and throughout lactation, and the fecal bacterial abundance and SCFA levels of the offspring were analyzed when they reached 12 months of age. Our results showed dose-dependent and sex-based differences in fecal microbial abundance at various taxonomic levels. Specifically, we found a decline in Firmicutes in males, and an increase in Actinobacteria among males and females. Furthermore, a sex-specific distribution reorganization of Lactobacillus, Bifidobacterium, and Akkermansia was identified. No significant difference in the concentration of prominent SCFAs and IgA levels were identified. These findings provide preliminary information indicating the need to evaluate perinatal effects of ART more comprehensively on the gut bacterial and metabolic function in future studies, and their potential role in offspring health outcomes.

Cannabis and cannabinoid mixtures have been linked to a variety of health benefits including pain mitigation, suppression of nausea produced by chemotherapeutic agents, anti-inflammatory effects, and effects on energy homeostasis, glucose, and lipid metabolism. The latter properties have led to the suggestion that these products could have therapeutic effects on the development of metabolic dysfunction-associated steatohepatitis (MASH) - a severe type of liver pathology in obese and diabetic patients. However, varying agonist and antagonistic properties of different cannabinoids on the endogenous cannabinoid system make prediction regarding hepatic effects and diet interactions difficult. The current study was designed to examine hepatic pathology following chronic administration of a cannabinoid mixture (NEPE14) at a dose equivalent to one previously demonstrating antihyperalgesic effects in rats. The effects of NEPE14 were investigated in a mouse model of MASH produced by feeding a Western diet rich in fat and simple sugars. After 24 weeks of NEPE14 administration, there was no hepatotoxicity in mice receiving the control diet and no significant exacerbation of MASH in mice receiving the Western diet. In conclusion, no chronic liver toxicity was observed, but there was also no evidence for protection against MASH by this product.