Electro-Oxidation and Simultaneous Determination of Indole-3-Acetic Acid and Salicylic Acid on Graphene Hydrogel Modified Electrode.

A selective and sensitive electrochemical sensor was developed for simultaneous detection of phytohormones indole-3-acetic acid (IAA) and salicylic acid (SA). The sensing interface was fabricated on a porous, three-dimensional networked graphene hydrogel (GH) modified glassy carbon electrode (GCE). The electrocatalytic behavior of IAA and SA on the surface of the modified electrode (GH/GCE) was investigated by cyclic voltammetry and linear sweep voltammetry. Results show that the oxidation reactions of IAA and SA occur at different potentials, which enable their simultaneous detection at the sensing interface. Under optimal conditions, the GH/GCE exhibited good selectivity and stability and its response, unaffected by various interferents, was linear in the range of 4 to 200 µM of IAA and SA. The limit of detection (S/N = 3) achieved were 1.42 µM for IAA and 2.80 µM for SA. The sensor performance was validated by measuring for IAA and SA in real vegetable samples with satisfactory results.

Update on HLA-B*15:02 allele associated with adverse drug reactions.

HLA alleles, part of the major histocompatibility complex, are strongly associated with adverse drug reactions (ADRs). This review focuses on HLA-B*15:02 and explores its association with ADRs in various ethnic populations and with different drugs, aiming to provide insights into the safe clinical use of drugs and minimize the occurrence of ADRs. Furthermore, the review explores the potential mechanisms by which HLA-B*15:02 may be associated with ADRs, aiming to gain new insights into drug modification and identification of haptens. In addition, it analyzes the frequency of the HLA-B*15:02, genotyping methods, cost-effectiveness and treatment measures for adverse reactions, thereby providing a theoretical basis for formulating clinical treatment plans.

Accurate identification of HLA-B*15:02 allele by two-dimensional polymerase chain reaction.

BACKGROUND: HLA-B*15:02 is highly associated with carbamazepine-induced SJS/TEN; however, there is no rapid and accurate detecting method. Here, we present a method to distinguish HLA-B*15:02 from 16 highly homologous HLA-B*15 alleles. METHODS: The high-throughput two-dimensional polymerase chain reaction (2D-PCR) technology was employed to identify HLA-B*15:02 in two-tube reaction. And, 2D-PCR accuracy was verified by PCR-sequence based typing (PCR-SBT). RESULTS: HLA-B*15:02 heterozygotes were identified by 14 melting valleys in the first tube reaction and none in the second, or by 13 melting valleys in the first tube reaction and one in the second. HLA-B*15:02 homozygote was identified by 13 melting valleys in the first tube reaction and none in the second. Three (0.16%) HLA-B*15:02 homozygotes and 84 (4.59%) HLA-B*15:02 heterozygotes were detected in 1830 samples of clinical general population without detecting 16 highly homologous alleles to HLA-B*15:02. The kappa test showed 100% coincidence between the 2D-PCR and PCR-SBT. CONCLUSIONS: 2D-PCR in two-tube reaction method for identifying HLA-B*15:02 was successfully established. Identification of HLA-B*15:02 is necessary prior to taking CBZ based on HLA-B*15:02 allele frequency.

Clusters of Body Fat and Nutritional Parameters are Strongly Associated with Diabetic Kidney Disease in Adults with Type 2 Diabetes.

INTRODUCTION: Diabetic kidney disease (DKD) has become the leading cause of chronic kidney disease and end-stage renal failure in most developed and many developing countries. Strategies aimed at identifying potential modifiable risk factors for DKD are urgently needed. Here, we investigated the association between clusters of body fat and nutritional parameters with DKD in adults with type 2 diabetes mellitus (T2DM). METHODS: This was a cross-sectional study of 184 participants with T2DM. Biochemical parameters including fasting blood glucose, hemoglobin A1c, hemoglobin, albumin, creatinine, and urinary albumin-to-creatinine ratio (UACR) were measured. The data for percentage of body fat mass (PBF), visceral fat area (VFA), phase angle at 50 kHz (PA50), and body cell mass (BCM) were obtained by bioelectrical impedance analysis (BIA). DKD was diagnosed by UACR and estimated glomerular filtration rate. Factor analysis was used for dimensionality reduction clustering among variables. The association of clusters with the presence of DKD was assessed using binary logistic regression analysis. RESULTS: Factor analysis identified two clusters which were interpreted as a body fat cluster with positive loadings of VFA, body mass index, waist circumstance, and PBF and a nutritional parameters cluster with positive loadings of PA50, hemoglobin, BCM, and albumin. Participants were divided into the four groups based on the sex-specific cutoff value (median) of each cluster score calculated using the cluster weights and the original variable values. Only participants with high body fat and poor nutritional parameters (OR 3.43, 95% CI 1.25-9.42) were associated with increased odds of having DKD. CONCLUSION: Body fat and nutritional parameters were strongly associated with and considerably contributed to the presence of DKD, suggesting that body fat and nutrition might be promising markers representing metabolic state in pathogenesis of DKD and clinical utility of BIA might provide valuable recommendations to patients with T2DM.

MscI restriction enzyme cooperating recombinase-aided isothermal amplification for the ultrasensitive and rapid detection of low-abundance EGFR mutations on microfluidic chip.

The detection of low-abundance mutation genes of the epidermal growth factor receptor (EGFR) exon 21 (EGFR L858R) plays a crucial role in the diagnosis of non-small cell lung cancer (NSCLC), as it enables early cancer detection and facilitates the development of treatment strategies. A detection platform was developed by combining the MscI restriction enzyme with the recombinase-aided isothermal amplification (RAA) technique (MRE-RAA). During the RAA process, "TGG^CCA" site of the wild-type genes was cleaved by the MscI restriction enzyme, while only the low-abundance mutation genes underwent amplification. Notably, when the RAA product was combined with CRISPR-Cas system, the sensitivity of detecting the EGFR L858R mutation increased by up to 1000-fold for addition of the MscI restriction enzyme. This achievement marked the first instance of attaining an analytical sensitivity of 0.001%. Furthermore, a disk-shaped microfluidic chip was developed to automate pretreatment while concurrently analyzing four blood samples. The microfluidic features of the chip include DNA extraction, MRE-RAA, and CRISPR-based detection. The fluorescence signal is employed for detection in the microfluidic chip, which is visible to the naked eye upon exposure to blue light irradiation. Furthermore, this platform has the capability to facilitate early diagnosis for various types of cancer by enabling high-sensitivity detection of low-abundance mutation genes.

Elimination of subtelomeric repeat sequences exerts little effect on telomere essential functions in Saccharomyces cerevisiae.

Telomeres, which are chromosomal end structures, play a crucial role in maintaining genome stability and integrity in eukaryotes. In the baker's yeast Saccharomyces cerevisiae, the X- and Y'-elements are subtelomeric repetitive sequences found in all 32 and 17 telomeres, respectively. While the Y'-elements serve as a backup for telomere functions in cells lacking telomerase, the function of the X-elements remains unclear. This study utilized the S. cerevisiae strain SY12, which has three chromosomes and six telomeres, to investigate the role of X-elements (as well as Y'-elements) in telomere maintenance. Deletion of Y'-elements (SY12YΔ), X-elements (SY12XYΔ+Y), or both X- and Y'-elements (SY12XYΔ) did not impact the length of the terminal TG1-3 tracks or telomere silencing. However, inactivation of telomerase in SY12YΔ, SY12XYΔ+Y, and SY12XYΔ cells resulted in cellular senescence and the generation of survivors. These survivors either maintained their telomeres through homologous recombination-dependent TG1-3 track elongation or underwent microhomology-mediated intra-chromosomal end-to-end joining. Our findings indicate the non-essential role of subtelomeric X- and Y'-elements in telomere regulation in both telomerase-proficient and telomerase-null cells and suggest that these elements may represent remnants of S. cerevisiae genome evolution. Furthermore, strains with fewer or no subtelomeric elements exhibit more concise telomere structures and offer potential models for future studies in telomere biology.

Urinary metabolites associate with the presence of diabetic kidney disease in type 2 diabetes and mediate the effect of inflammation on kidney complication.

AIMS: Diabetic kidney disease (DKD) is the one of the leading causes of end-stage kidney disease. Unraveling novel biomarker signatures capable to identify patients with DKD is favorable for tackle the burden. Here, we investigated the possible association between urinary metabolites and the presence of DKD in type 2 diabetes (T2D), and further, whether the associated metabolites improve discrimination of DKD and mediate the effect of inflammation on kidney involvement was evaluated. METHODS: Two independent cohorts comprising 192 individuals (92 DKD) were analyzed. Urinary metabolites were analyzed by targeted metabolome profiling  and inflammatory cytokine IL-18 were measured by ELISA. Differentially expressed metabolites were selected and mediation analysis was carried out. RESULTS: Seven potential metabolite biomarkers (i.e., S-Adenosyl-L-homocysteine, propionic acid, oxoadipic acid, leucine, isovaleric acid, isobutyric acid, and indole-3-carboxylic acid) were identified using the discovery and validation design. In the pooled analysis, propionic acid, oxoadipic acid, leucine, isovaleric acid, isobutyric acid, and indole-3-carboxylic acid were markedly and independently associated with DKD. The composite index of 7 potential metabolite biomarkers (CMI) mediated 32.99% of the significant association between the inflammatory IL-18 and DKD. Adding the metabolite biomarkers improved the discrimination of DKD. CONCLUSIONS: In T2D, several associated urinary metabolites were identified to improve the prediction of DKD. Whether interventions aimed at reducing CMI also reduce the risk of DKD especially in patients with high IL-18 needs further investigations.

Survey and toxigenic abilities of Aspergillus, Fusarium, and Alternaria fungi from wheat and paddy grains in Shanghai, China.

A systematic study was carried out on 638 wheat and paddy grains (including fresh and stored samples) collected in 2021 from Shanghai, China, to identify the major mycobiota and their toxigenic abilities. A total of 349 fungi, namely, 252 Fusarium, 53 Aspergillus, and 44 Alternaria, were characterized by morphological and molecular identification. Fusarium and Aspergillus were more frequently isolated in paddy with Fusarium sambucinum species complex and Aspergillus section flavi as the predominant species, respectively. The genus Alternaria was the most frequently isolated fungal species in wheat. The toxin-producing potentials of the identified fungi were further evaluated in vitro. Deoxynevalenol (DON) was produced by 34.5% of Fusarium isolates and zearalenone (ZEN) was produced by 47.6% of them, and one isolate also processed the abilities for fumonisin B1 (FB1), B2 (FB2), and B3 (FB3) productions. Aflatoxin B1 (AFB1), B2 (AFB2), and G1 (AFG1) were only generated by Aspergillus section flavi, with the production rate of 65.5%, 27.6%, and 13.8%, respectively. Alternariol (AOH) was the most prevalent Alternaria toxin, which could be produced by 95.5% of the isolates, followed by alternariol monomethyl ether (AME) (72.7%), altenuene (ALT) (52.3%), tenuazonic acid (TeA) (45.5%), tentoxin (TEN) (29.5%), and altenusin (ALS) (4.5%). A combinational analysis of mycobiota and toxigenic ability allowed us to provide comprehensive information about the production mechanisms of mycotoxins in wheat and paddy in a specific geographic area, and will be helpful for developing efficient prevention and control programs.

Urinary IL-18 is associated with arterial stiffness in patients with type 2 diabetes.

Objective: Diabetic kidney disease (DKD) has been shown to be associated with an excess risk of cardiovascular death. Inflammation has been considered central to type 2 diabetes (T2D) pathophysiology, and inflammation markers have been linked to cardiovascular disease. The serum and urinary IL-18 levels were significantly elevated in patients with T2D; however, whether interleukin 18 (IL-18) are associated with the severity of arterial stiffness remains to be determined. This study examined the relationship of IL-18 levels with pulse wave velocity (PWV) as a reflector for arterial stiffness in patients with T2D. Methods: A total of 180 participants with T2D who had undergone PWV examination were enrolled. Serum and urinary IL-18 levels were measured using sandwich enzyme linked immunosorbent assay (ELISA) kits. Arterial stiffness was determined by carotid-femoral PWV (cf-PWV) and carotid-radial PWV (cr-PWV). Results: The urinary IL-18 levels correlated positively with cf-PWV in patients with T2D with DKD (r = 0.418, p < 0.001); however, we found no significant correlation between urinary IL-18 and cf-PWV in diabetic subjects without DKD. In addition, we found no significant correlation between urinary IL-18 and cr-PWV in participants with T2D with or without DKD. Moreover, the association remained significant when controlling for arterial stiffness risk factors, urinary albumin-to-creatinine ratio and estimated glomerular filtration rate. cf-PWV was greater in the higher group of urinary IL-18 than in the lower group. Nevertheless, we found no significant correlation between serum IL-18 and cf-PWV in participants with T2D. Conclusion: The urinary IL-18 levels appear to be associated with greater cf-PWV, suggesting the link between urinary IL-18 and arterial stiffness in patients with T2D.

JNK/AP1 Pathway Regulates MYC Expression and BCR Signaling through Ig Enhancers in Burkitt Lymphoma Cells.

In Burkitt lymphoma (BL), a chromosomal translocation by which the MYC gene is fused to an immunoglobulin (Ig) gene locus is frequently found. The translocated MYC gene is overexpressed, which is the major driver of BL tumorigenesis. Studies have shown that Ig enhancers are essential for MYC overexpression, but the involved mechanisms are not fully understood. In addition, the survival of BL cells relies on B-cell receptor (BCR) signaling, which is determined by the levels of Ig molecules expressed on the cell surface. However, whether MYC has any impact on Ig expression and its functional relevance in BL has not been investigated. Herein, we show that MYC upregulates Ig kappa (Igκ) expression in BL cells through two Igκ enhancers, the intronic enhancer (Ei) and the 3' enhancer (E3'). Mechanistically, by activating the JNK pathway, MYC induces the phosphorylation of c-Fos/c-Jun and their recruitment to AP1 binding sites in the Igκ enhancers, leading to the activation of the enhancers and subsequent Igκ upregulation. The AP1-mediated activation of the Igκ enhancers is also required for the expression of the translocated MYC gene, indicating positive feedback for the MYC overexpression in BL cells. Importantly, interrupting the JNK pathway inhibits both Igκ and MYC gene expression and suppresses BL cell proliferation. Our study not only reveals a novel mechanism underlying MYC overexpression in BL but also suggests that targeting the JNK pathway may provide a unique strategy to suppress BL tumorigenesis.

Gold nanoparticle-doped three-dimensional reduced graphene hydrogel modified electrodes for amperometric determination of indole-3-acetic acid and salicylic acid.

Three-dimensional (3D) networked nanomaterials have attracted great interest because of their unique porous and 3D-networked structures. In this work, a series of gold nanoparticle (AuNP) doped graphene hydrogel nanocomposites (AuNP-GHs) were synthesized through hydrothermal reaction under various conditions. X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS) were used to characterize the AuNP-GH. The AuNP-GH was used to modify the glassy carbon electrode (GCE) for the detection of indole-3-acetic acid (IAA) and salicylic acid (SA) using chronoamperometric measurements. Under optimum conditions, the AuNP-GH/GCE exhibited linear response to IAA in the ranges of 0.8-4 µM and 4-128 µM, and to SA in the ranges of 0.8-8.4 µM and 8.4-188.4 µM. The detection limits (S/N = 3) were calculated to be 0.21 µM for IAA and 0.22 µM for SA. The proposed sensor showed good sensitivity and stability and hence it was applied in the detection of IAA and SA in spiked samples with satisfactory results.

miR-27a-containing Exosomes Secreted by Irradiated Skin Keratinocytes Delayed the Migration of Unirradiated Skin Fibroblasts.

Radiation-induced bystander effect (RIBE), e.g. the biological response occurring in unirradiated cells when their neighboring cells are irradiated, is the consequence of intercellular communication between irradiated and unirradiated cells and intracellular signal transduction of these two cell populations. Although several miRNAs have been found to play an important role in RIBEs, the evidence for the regulatory effects of miRNAs on RIBEs is still limited. In this study, by using a two cell-line co-culture system, we first found that the migration of unirradiated bystander WS1 skin fibroblasts was inhibited after co-culture with irradiated HaCaT skin keratinocytes. Further study revealed that HaCaT cells exposed to α-particles and X-rays quickly showed an elevated miR-27a expression, which was essential for the induction of the bystander effect, resulting in the secretion of miR-27a-containing exosomes as a major RIBE signaling factor. Upon uptake of these exosomes, the recipient unirradiated WS1 cells displayed oxidative stress and increased miR-27a levels. Elevated levels of miR-27a that targets MMP2 in the recipient WS1 cells then led to slowed cell migration, which was dependent upon the redox status of WS1 cells. To summarize, the present study has revealed a critical role of miR-27a in every step of the induction of bystander migration inhibition of unirradiated WS1 fibroblasts co-cultured with irradiated HaCaT keratinocytes, confirming the important regulatory effects of miRNAs in RIBEs. Additionally, we provided direct evidence that RIBEs could affect wound healing.