Search

In mild cases, it is difficult to diagnose pancreatic exocrine insufficiency (PEI). There is no gold standard method for the diagnosis of PEI. A reliable method is needed for preliminary diagnosis of PEI. The PEI-TEST was applied to the patients with nonspecific gastrointestinal complaints. Serum amylase, lipase, serum trypsinogen, and fecal elastase 1 (FE-1) were analyzed from each patient. According to the PEI-TEST, PEI was present in 42 (47.7%) and PEI was not present in 46 (52.3%) patients. No significant difference was observed between the 2 groups with regard to age, gender and amylase, lipase, serum trypsinogen, and FE-1. When an FE-1 value of <200 µg/dL was considered as indicating PEI, the sensitivity and specificity of the test were found to be 47.4% and 52.2%, respectively. Although it is promising that PEI-TEST is a validated test in our country and suitable for our society, it is not suitable for pediatric patients. [ABSTRACT FROM AUTHOR]

Background: The oral cavity is a complex environment which harbours the second largest and most diverse microflora after the gastrointestinal tract. The bacteriome in the oral cavity plays a pivotal role in promoting the health and well-being of human beings. Gingivitis, an inflammation of the gingival tissue, arises due to plaque accumulation on the teeth, often leads to periodontitis. Progression of periodontitis resulting in clinical attachment loss, bone loss and eventually the tooth loss is poorly understood. The present study explores the transitions in microbioata, oxidative stress and polyamine levels during the disease evolution which can contribute to developing effective therapeutic approaches. Methods: Saliva samples were collected from seventy-two individuals after procuring informed consent who were either healthy, gingivitis or stage-specific periodontitis patients. Periodontitis stage was confirmed by clinical and radiographic analysis. Microbiota analysis was carried out by 16S rRNA sequencing on the Nanopore PromethIONsystem platform of Oxford Nanopore technologies. Polyamine levels were quantified with fluorescence spectrophotometer. Ornithine decarboxylase quantification was evaluated by ELISA method. Antioxidant levels of the salivary samples were measured by DPPH, SOD, and catalase assays. Autophagy was measured by acid phosphatase assay. Result: The salivary microbiota exhibited significant changes in their abundance and diversity between healthy individuals and those with conditions such as gingivitis, and chronic periodontitis. A significant increase in polyamines and ornithine decarboxylase was found in gingivitis and various stages of periodontitis. Elevated oxidative stress observed in gingivitis and periodontitis could have resulted in cell death. Conclusion: The current study shows the role of salivary microbiota and polyamines in gingivitis and different periodontitis stages. The progressive elevation of Streptococcus levels from gingivitis to periodontitis, coupled with polyamine concentrations, may serve as a promising identification marker for assessing the severity of periodontal disease. Insight into the oral bacterial flora and associated physiological changes provide a foundation for targeted therapeutic interventions in gingivitis and periodontitis diseases emphasising the importance of personalised oral health management strategies. [ABSTRACT FROM AUTHOR]

Background: Periodontal diseases are chronic inflammatory conditions that require early screening for effective long‐term management. Oral neutrophil counts (ONCs) correlate with periodontal inflammation. This study investigates a point‐of‐care test using a neutrophil enzyme activity (NEA) colorimetric strip for measuring periodontal inflammation. Methods: This prospective study had two phases. Phase 1 validated the relationship between ONCs and periodontal inflammation with 90 participants. Phase 2 examined the test's applicability in a real‐world setting through a multicentre clinical trial with 375 participants at four sites. ONCs were quantified in oral rinses using laboratory‐based methods, and the NEA strip was used for ONC stratification. Clinical measures included bleeding on probing (BoP), probing depth (PD) and clinical attachment loss (CAL). Results: ONCs were significantly elevated in patients with Grade B periodontitis and deep periodontal pockets (PD ≥ 5 mm, CAL ≥ 5 mm). The NEA strip accurately classified patients into high or low ONC categories, showing 80% sensitivity, 82.5% specificity and an AUC of 0.89. It also assessed the effectiveness of periodontal therapy in reducing ONC and inflammation. The test was user‐friendly, with no reported discomfort among patients. Conclusion: The NEA strip is a user‐friendly and rapid screening tool for detecting high ONCs associated with periodontal inflammation and for evaluating the effectiveness of periodontal therapy. [ABSTRACT FROM AUTHOR]

Simple Summary: PTEN is an essential tumor suppressor gene whose alterations are causative of cancer and neurodevelopmental disorders, grouped as PTEN hamartoma tumor syndrome (PHTS). Here, we present translational readthrough as a potential therapeutic approach to restore the function of PTEN isoforms in patients harboring nonsense mutations at PTEN that cause aberrant PTEN biosynthesis. We have found that aminoglycosides and protein synthesis inducers reconstitute PTEN isoform expression and the function of PTEN variants containing nonsense mutations. Our results provide insights with which to optimize readthrough-based therapies for patients with cancer and PHTS. The PTEN tumor suppressor is frequently targeted in tumors and patients with PTEN hamartoma tumor syndrome (PHTS) through nonsense mutations generating premature termination codons (PTC) that may cause the translation of truncated non-functional PTEN proteins. We have previously described a global analysis of the readthrough reconstitution of the protein translation and function of the human canonical PTEN isoform by aminoglycosides. Here, we report the efficient functional readthrough reconstitution of the PTEN translational isoform PTEN-L, which displays a minimal number of PTC in its specific N-terminal extension in association with disease. We illustrate the importance of the specific PTC and its nucleotide proximal sequence for optimal readthrough and show that the more frequent human PTEN PTC variants and their mouse PTEN PTC equivalents display similar patterns of readthrough efficiency. The heterogeneous readthrough response of the different PTEN PTC variants was independent of the length of the PTEN protein being reconstituted, and we found a correlation between the amount of PTEN protein being synthesized and the PTEN readthrough efficiency. Furthermore, combination of aminoglycosides and protein synthesis inducers increased the readthrough response of specific PTEN PTC. Our results provide insights with which to improve the functional reconstitution of human-disease-related PTC pathogenic variants from PTEN isoforms by increasing protein synthesis coupled to translational readthrough. [ABSTRACT FROM AUTHOR]

Background: Alzheimer’s disease (AD) is a complex neurodegenerative disorder influenced by various factors, including liver function, which may impact the clearance of amyloid-β (Aβ) in the brain. This study aimed to explore how the apolipoprotein E (APOE) ε4 allele affects the relationship of liver function markers with AD pathology and cognition. Methods: We analyzed data from two independent cohorts, including 732 participants from the Hallym University Medical Center and 483 from the Alzheimer’s Disease Neuroimaging Initiative, each group consisting of individuals with and without the APOE ε4 allele. Cross-sectional analyses evaluated the associations of liver enzymes (aspartate aminotransferase [AST], alanine aminotransferase [ALT], alkaline phosphatase, total bilirubin, and albumin) with AD diagnosis, amyloid positron emission tomography (PET) burden, and cerebrospinal fluid biomarkers for AD (Aβ42, total tau, and phosphorylated tau181) at baseline. Longitudinally, we investigated the associations between these liver enzymes and changes in cognitive performance over the course of a year. Logistic and linear regression models were used to analyze these associations and mediation analyses were conducted to assess whether age and amyloid PET burden mediated these associations. Results: Only in the APOE ε4 carrier group, a high AST to ALT ratio and low ALT levels were significantly associated with AD diagnosis, increased amyloid PET burden, and faster longitudinal decline in cognitive function in both cohorts. In particular, the AST to ALT ratio was associated with cerebrospinal fluid Aβ42 levels exclusively in the APOE ε4 carrier group in the Alzheimer’s Disease Neuroimaging Initiative cohort but not with phosphorylated tau181 or total tau levels. Moreover, mediation analyses from both cohorts revealed that in the APOE ε4 carriers group, age did not mediate the associations between liver enzymes and AD diagnosis or amyloid PET burden. However, amyloid PET burden partially mediated the association between liver enzymes and AD diagnosis exclusively in the APOE ε4 carriers group. Conclusion: This study provides valuable insights into the significant association of the APOE ε4 allele with liver enzymes and their potential role in Aβ-related pathogenesis and cognition in AD. Further research is required to elucidate the underlying mechanisms and potential therapeutic implications of these findings. [ABSTRACT FROM AUTHOR]

Simple Summary: In breast cancer (BC), the triple-negative breast cancer subtype, the upregulation of CDC20 is positively associated with cancer initiation, progression, metastasis, and chemotherapy resistance. The current functional enrichment analysis study demonstrated the significant association of CDC20 co-expressed genes with biological regulation and cellular processes. This study reveals a significant positive correlation between overexpressed CDC20 and tumor purity and many immune cells; the finding suggests that CDC20 plays a fundamental role in controlling tumor immunity and consequently influences BC prognosis. CDC20 deficiency led to decreased cell growth and metastasis, G2/M cell cycle arrest, and boosted the cytotoxic effects of paclitaxel treatment, which supports the current analysis. Developing natural and synthetic inhibitors of this oncogene is a promising approach to the therapeutic management of BC. Cell division cycle 20 homolog (CDC20) is a well-known regulator of cell cycle progression. Abnormal expression of CDC20 leads to mitotic defects, which play a significant role in cancer development. In breast cancer (BC), CDC20 has been identified as a biomarker that has been linked to poor patient outcomes. In this study, we investigated the association of CDC20 with BC prognosis and immune cell infiltration by using multiple online databases, including UALCAN, KM plotter, TIMER2.0, HPA, TNM-plot, bc-GenExMiner, LinkedOmics, STRING, and GEPIA. The results demonstrate that BC patients have an elevated CDC20 expression in tumor tissues compared with the adjacent normal tissue. In addition, BC patients with overexpressed CDC20 had a median survival of 63.6 months compared to 169.2 months in patients with low CDC20 expression. Prognostic analysis of the examined data indicated that elevated expression of CDC20 was associated with poor prognosis and a reduction of overall survival in BC patients. These findings were even more prevalent in chemoresistance triple-negative breast cancer (TNBC) patients. Furthermore, the Gene Set Enrichment Analysis tool indicated that CDC20 regulates BC cells' cell cycle and apoptosis. CDC20 also significantly correlates with increased infiltrating B cells, CD4+ T cells, neutrophils, and dendritic cells in BC. In conclusion, the findings of this study suggest that CDC20 may be involved in immunomodulating the tumor microenvironment and provide evidence that CDC20 inhibition may serve as a potential therapeutic approach for the treatment of BC patients. In addition, the data indicates that CDC20 can be a reliable prognostic biomarker for BC. [ABSTRACT FROM AUTHOR]

Background: Extended contact with silica particles can lead to Silicosis, a chronic lung condition lacking established treatment protocols or clear mechanisms of development. The urgency for innovative treatments arises from the unavailability of effective treatment methodologies. The origin of silica-induced pulmonary fibrosis includes essential processes such as macrophage activation and the conversion of fibroblasts into myofibroblasts, with oxidative stress playing a pivotal role. Shionone (SHI), a triterpenoid extracted from the Aster tataricus plant, is recognized for its extensive health benefits. This study explores the capability of SHI to alleviate the effects of silica-induced lung fibrosis in mice. Methods: This investigation explored the impact of SHI on lung inflammation and fibrosis at different stages (early and late) triggered by silica in mice, focusing specifically on the initial and more developed phases. It comprised an analysis of isolated peritoneal macrophages and fibroblasts extracted from mice to elucidate SHI's therapeutic potential and its underlying mechanism. The methodology employed encompassed quantitative PCR, immunofluorescence, flow cytometry, and western blotting to examine macrophage activity and their transition into myofibroblasts. The activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway by SHI was confirmed via immunofluorescence and western blot studies. SHI's antioxidative properties were evidenced by the measurement of reactive oxygen species (ROS) and mitochondrial ROS within both macrophages and fibroblasts, using 2′, 7′-dichlorodihydrofluorescein diacetate and MitoSOX, respectively. The relevance of SHI was further underscored by applying ML385 and Nrf2 siRNA to gauge its effectiveness. Results: Starting SHI treatment early countered the harmful effects of lung inflammation and fibrosis caused by silica, while initiating SHI at a later phase decelerated the advancement of fibrosis. SHI's action was linked to the activation of the Nrf2 signaling pathway, a boost in antioxidant enzyme levels, and a decrease in oxidative stress and inflammation in macrophages affected by silica. Furthermore, SHI prevented the conversion of fibroblasts into myofibroblasts prompted by TGF-β, along with the resultant oxidative stress. The beneficial outcomes of SHI were negated when ML385 and Nrf2 siRNA were applied, highlighting the pivotal role of the Nrf2 pathway in SHI's efficacy. Conclusion: SHI plays a significant role in stimulating the Nrf2 pathway, thereby defending against silica-induced oxidative stress and inflammatory reactions in macrophages, and inhibiting the conversion of fibroblasts to myofibroblasts due to TGF-β. This suggests that SHI is a viable option for treating lung inflammation and fibrosis in mice suffering from silicosis. [ABSTRACT FROM AUTHOR]

Background: This study aimed to validate alterations in the gene expression of DNA methylation-related enzymes and global methylation in the peripheral blood mononuclear cell (PBMC) and synovial tissues of animal hip osteoarthritis (OA) models. Methods: Animals were assigned to the control (no treatment), sham (25 µL of sterile saline), and OA (25 µL of sterile saline and 2 mg of monoiodoacetate) groups. Microcomputed tomography scan, histopathological assessment and pain threshold measurement were performed after induction. The mRNA expression of the DNA methylation machinery genes and global DNA methylation in the PBMC and hip synovial tissue were evaluated. Results: The OA group presented with hip joint OA histopathologically and radiologically and decreased pain threshold. The mRNA expression of DNA methyltransferase (Dnmt 3a), ten–eleven translocation (Tet) 1 and Tet 3 in the synovial tissue of the OA group was significantly upregulated. Global DNA methylation in the synovial tissue of the OA group was significantly higher than that of the control and sham groups. Conclusions: The intra-articular administration of monoiodoacetate induced hip joint OA and decreased pain threshold. The DNA methylation machinery in the synovial tissues of hip OA was altered. [ABSTRACT FROM AUTHOR]

A 60-day feeding trial was conducted to evaluate the effect of papaya (Carica papaya) leaf meal on the physio-metabolic enzyme response of Labeo rohita fingerlings. Fingerlings were distributed in five different experimental groups with replicates. Each replicate contains 15 fishes with a mean average initial weight of 11.34 ± 0.20 g-11.61 ± 0.04 g and is fed daily twice for 60 days. Five practical iso-nitrogenous fish diets (~30% protein) were prepared with different levels of papaya leaf meal (PLM) with partial replacement of rice bran, that is T1 (0% PLM), T2 (10% PLM), T3 (15% PLM), T4 (20% PLM) and T5 (25% PLM). The digestive enzymes, such as amylase and protease varied significantly. Amylase activity was significantly higher in the T3 group, while protease activity was highest in the T4 group. The activities of metabolic enzymes, such as alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and malate dehydrogenases were lowest in the T4 and T5 groups. The oxidative stress enzymes were also recorded for their minimum values in the T5 group. The overall results indicate that the fish fed with 20% PLM (T4 group) showed the best performance in terms of growth and physio-metabolic enzyme parameters, although some of the parameters indicated a better value at 25% inclusion. Hence, it can be concluded that PLM can be included up to 20% in the diet of L. rohita (Rohu) fingerlings for optimum production without any deleterious effect on enzyme activity, which is mostly required for fish growth and production. [ABSTRACT FROM AUTHOR]

Simple Summary: As the precursor of pepsin, Pepsinogen C (PGC) is the final product of the maturation and differentiation of gastric mucosa cells. The latest bioinformatics analysis found that PGC may play a little-known role in the carcinogenic process, but there is no relevant experimental evidence to prove the effect of PGC on the biological function of cancer cells. In our research, we found PGC may act as a tumor suppressor in the development and metastasis of gastric cancer. PGC can downregulate its interacting protein IQGAP1 and inhibit the Rho-GTPase pathway, thereby participating in the inhibition of gastric cancer cell migration and invasion. Thus, PGC has the potential to be a new therapeutic target for gastric cancer. Aim: This study systematically explored the biological effects and mechanisms of PGC on gastric cancer (GC) cells in vitro and in vivo. Method: The critical biological roles of PGC in GC were assessed via EdU staining, Hoechst staining, flow cytometry, mouse models, CCK-8, wound healing, transwell, and sphere-forming assays. The interaction study with IQ-domain GTPase-activating protein 1 (IQGAP1) was used by Liquid chromatography-mass spectrometry co-immunoprecipitation, immunofluorescence staining, CHX-chase assay, MG132 assay, and qRT-PCR. Results: PGC inhibited the proliferation, viability, epithelial–mesenchymal transition, migration, invasion, and stemness of GC cells and promoted GC cell differentiation. PGC suppressed subcutaneous tumor growth and peritoneal dissemination in vivo. The interaction study found PGC inhibits GC cell migration and invasion by downregulating IQGAP1 protein and IQGAP1-mediated Rho-GTPase signaling suppression. In addition, PGC disrupts the stability of the IQGAP1 protein, promoting its degradation and significantly shortening its half-life. Moreover, the expression levels of PGC and IQGAP1 in GC tissues were significantly negatively correlated. Conclusion: PGC may act as a tumor suppressor in the development and metastasis of GC. PGC can downregulate its interacting protein IQGAP1 and inhibit the Rho-GTPase pathway, thereby participating in the inhibition of GC cell migration and invasion. [ABSTRACT FROM AUTHOR]

Stenotrophomonas koreensis, was isolated from a textile effluent and employed to biologically degrade 98.8% of the recalcitrant toxic dye malachite green in 4 h in the absence of any supplements or media. Chromatographic and Spectroscopic analysis confirmed the degradation of the dye. According to Response Surface Methodology, the optimum conditions for biodegradation were pH 6, inoculum size 6 mL (initial concentration Log10CFU/mL = 6.2), and temperature 45 °C. More than 98% decolorization was achieved within 2.5 h of the addition of soymeal extract or peptone. One of the most promising features of this strain is that even in the presence of heavy metals, S. koreensis actively degraded the dye. The bacteria biodegraded malachite green following the first-order reaction kinetics. LC-MS analysis of the degradation product yielded several intermediates like Michler's ketone (m/z 269), 4-N,N-dimethylaminophenol (m/z 137), Benzophenone (m/z 182), N,N-dimethylaniline (m/z 121), 4-(N-methylamino)-benzophenone (m/z 211), 4-aminobenzaldehyde (m/z 121), anionic canonical form of 4-aminobenzaldehyde (m/z 120), 4-(N,N-dimethylamino)benzoic acid (m/z 165), 4-(N-methylamino) benzoic acid (m/z 152). The enzymes responsible for the malachite green degradation were tyrosinase, Malachite Green reductase, and NADH-DCIP reductase. The present work is the first to report the degradation of malachite green by S. koreensis. [ABSTRACT FROM AUTHOR]

Aim: Preeclampsia (PE) is a pregnancy‐specific syndrome characterized by hypertension and proteinuria. Recently, multiple circular RNAs (circRNAs) were considered latent clinical diagnostic markers or therapeutic targets. This study was to explore the impact of circRNA serum and glucocorticoid‐induced kinase 1 (SGK1) on PE via influencing the microRNA (miR)‐508‐3p/PUM homolog 1 (PUM1) axis. Methods: Placental tissues of 34 pregnant women with PE and 34 normal pregnant women were collected to measure circRNA SGK1 (circSGK1), miR‐508‐3p, and PUM1. Human placental trophoblasts HTR‐8/SVneo were transfected with plasmids, thereafter to observe proliferation, migration, invasion, and apoptosis, analyze antioxidant molecules Troxerutin (TXN), Glutamate‐cysteine ligase catalytic subunit (GCLC), NAD (P) H‐quinone oxidoreductase 1 (NQO1), and determine angiogenesis. After the construction of the PE rat model, antioxidant molecules TXN, GCLC, and NQO1, vascular‐associated factor vascular endothelial growth factor A (VEGF‐A), and histopathological conditions were tested. Examination of the binding of circSGK1 and miR‐508‐3p with PUM1 was performed. Results: Our data showed that circSGK1 expression was elevated in the placenta of patients with PE. Silenced circSGK1 or elevated miR‐508‐3p promoted the growth and antioxidant molecules and angiogenesis in trophoblast cells; CircSGK1 combined with miR‐508‐3p, and miR‐508‐3p targeted PUM1. Conclusions: In summary, suppression of circSGK1 augments antioxidant molecules and angiogenesis in trophoblast cells to attenuate PE via miR‐508‐3p to target PUM1. [ABSTRACT FROM AUTHOR]

Agricultural pests can be effectively controlled using microbes, providing an eco-friendly alternative to available synthetic pesticides. Suitable entomopathogenic bacterial strains were collected from agricultural fields and evaluated for their insecticidal potential against Cnaphalocrocis medinalis. In the four tested entomopathogenic bacteria (W1, Yc1, S1, EB01), the larval mortality ranged from 38 to 74%. Among these isolates, Bacillus subtilis (EB01) induced the highest mortality (74%). In greenhouse conditions, the tests confirm that the results were dosage-dependent: B. subtilis infection considerably delayed the overall development period, reduced pupal conversion, and decreased adult emergence with induced morphological deformities. Larvae fed B. subtilis-treated leaves initiate bacterial infection and broadly damage the midgut tissue, including the epithelial and peritrophic layers. The bacterial growth in the C. medinalis hemolymph considerably increases the activity of enzymes like α and β esterase (85.14 and 44% at 96 h) compared to the control. The isolate B. subtilis-treated diet significantly reduced the larval digestive α and β galactosidase enzyme activity (88.17 and 91.88% at 96 h). Furthermore, germination bioassay with strain EB01 in rice varieties (TN1 and ASD16) significantly increased both varieties' germination and biomass index. This study shows that the B. subtilis EB01 strain potentially inhibited the biological activity of C. medinalis and improved the rice seeds' germination index. It can be a potential biocontrol agent in sustainable pest-management strategies. [ABSTRACT FROM AUTHOR]

The principal motive of this study is to explore the influence maternal separation (MS) exhibits on the mRNA expression of major drug metabolizing-cyp450s in parallel with the assessment of pathological changes that can be induced by MS in the livers of experimental mice. Eighteen Balb/c mouse pups, comprising of both males and females, were separated from their mothers after birth. Following a six-week period during when the pups became adults, the mice were sacrificed and their livers were isolated for analysis of weight, pathohistological alterations, and the mRNA expression of drug metabolizing cyp450 genes: cyp1a1, cyp3a11, cyp2d9, and cyp2c29. The study demonstrated that MS markedly downregulated (p<0.05) the mRNA expression of all tested drug-metabolizing cyp450s in livers of female and male mice. Furthermore, the mRNA levels of major drug-metabolizing cyp450s were notably lower (p<0.05) in livers of female MS mice as compared with male MS mice. It was found that values of the total body weight and liver weight of MS mice did not vary significantly (p>0.05) from those of the control groups. Additionally, histological examination revealed that the hepatic tissue of MS mice was normal, similar to that of the control mice. In summary, MS downregulates the gene expression of major hepatic drug-metabolizing cyp450s without inducing pathological alterations in the livers of mice. These findings provide an explanation for the heterogeneity in pharmacokinetics and drug response of patients with early life stress. [ABSTRACT FROM AUTHOR]

Background: The optimal conditioning regimen of tandem high-dose chemotherapy (HDC) and autologous stem cell transplantation (ASCT) for high-risk neuroblastoma (HR-NBL) has not been established. The efficacy of 131I-MIBG therapy is under exploration in newly diagnosed HR-NBL patients. Here, we compared the outcomes of tandem HDC/ASCT between the 131I-MIBG combination and non-MIBG groups. Methods: We retrospectively analyzed the clinical data of 33 HR-NBL patients who underwent tandem HDC/ASCT between 2007 and 2021 at the Seoul National University Children's Hospital. Results: The median age at diagnosis was 3.6 years. 131I-MIBG was administered to 13 (39.4%) of the patients. Thirty patients (90.9%) received maintenance therapy after tandem HDC/ASCT, twenty-two were treated with isotretinoin ± interleukin-2, and eight received salvage chemotherapy. The five-year overall survival (OS) and event-free survival (EFS) rates of all patients were 80.4% and 69.4%, respectively. Comparing the 131I-MIBG combined group and other groups, the five-year OS rates were 82.1% and 79.7% (p = 0.655), and the five-year EFS rates were 69.2% and 69.6% (p = 0.922), respectively. Among the adverse effects of grade 3 or 4, the incidence of liver enzyme elevation was significantly higher in the non-131I-MIBG group. Conclusions: Although tandem HDC/ASCT showed promising outcomes, the 131I-MIBG combination did not improve survival rates. [ABSTRACT FROM AUTHOR]

Aim. Among the methods of improving biosensor systems, the technique of using multi-enzyme bioselective elements is actively used. Its main idea is that, in addition to the main enzyme, auxiliary enzymes are added to the biosensor membrane, which can improve analytical characteristics or enhance the response of the biosensor to the target analyte. As a model biosensor that we intended to improve, we chose a conductometric biosensor for glucose determination based on glucose oxidase. The main enzyme of this biosensor was glucose oxidase (GOx), which is an enzyme of the oxidase class and catalyzes the reaction of splitting glucose into glucono-1,5-lactone and hydrogen peroxide using oxygen during the reaction. As an auxiliary enzyme, we additionally added catalase (CAT) to improve the characteristics of the sensor. It decomposes the peroxide formed during the first reaction with the participation of GOx into water and O2. This enables to increase the concentration of oxygen in the near-electrode layer, which improves the work of the main enzyme — GOx, and, accordingly, the entire biosensor. Methods. Two enzyme gels, glucose oxidase (GOx) and catalase (CAT) were mixed in a 2:1 ratio and applied to the first pair of electrodes for enzyme immobilization. A BSA protein gel was applied to the second pair of electrodes for reference. Immobilization occurred in glutaraldehyde vapors for 20 minutes, followed by air-drying for 10 minutes. The sensor was then washed for 5 minutes to remove residual GA and other unbound elements. Measurements were conducted in a 2 ml open cell with a 5 mM phosphate buffer solution at pH 6.2. The substrate concentration was adjusted by adding aliquotes of 500 mM D-glucose to working solution. Response time was 2–3 minutes with a 5-minute wash between responses. Response measurement could be done by comparing baseline conductance before and after exposure to the target molecule. Results and Discussion. The initial step in developing of the GOx-CAT bienzyme biosensor was to identify the most effective enzyme immobilization method on the transducer surface. We tested five biosensor variants based on different coimmobilization techniques of GOx and CAT and assessed their key analytical properties. Calibration curves were also generated for each variant to guide the selection process. Ultimately, experiments revealed that immobilization in glutaraldehyde vapors, combining GOx and CAT in a 2:1 ratio within a single-layer membrane, offered the highest sensitivity and wide linear operating range. This method was thus chosen for further development. The next stage of the work was the selection of the optimal concentration of catalase in the membrane. For this, a number of biosensors with different concentrations of catalase in the composition of the bioselective element were made and the sensitivity of these biosensors to the target analyte was compared. Thus, it was shown that the biosensor based on 5% CAT was characterized by the greatest response to the substrate. The difference between mono- and bienzyme biosensors lies in their sensitivity to glucose and their linear detection range. The sensitivity of the bienzyme biosensor (CAT-GOx mixture) was 378 μS/mM, compared to 161 μS/mM for the monoenzyme biosensor (GOx alone). Adding CAT to the biosensor extended its linear range to 1700 μM, compared to 800 μM for the monoenzyme biosensor. Additionally, the minimum detection limit decreased from 16 μM to 8 μM glucose for the monoenzyme and bienzyme biosensors, respectively. Conclusions. The study showed that the addition of catalase as an auxiliary enzyme significantly improved the analytical characteristics of the biosensor based on GOx, in particular, sensitivity, repeatability of measurement results, limits of linear and dynamic ranges of operation. It has been proven that the technology of adding an additional enzyme to the biomembrane of a biosensor can be used to improve the analytical characteristics of biosensors based on oxidases, which in turn will allow their more effective use. [ABSTRACT FROM AUTHOR]

Aedes albopictus (Skuse, 1895) (Diptera: Cullicidae) is an aggressive and invasive mosquito species. It has been shown that Ae. albopictus, which can be a vector in spreading some diseases such as zika, dengue, and chikungunya, develops resistance to some insecticides. This study investigated enzyme level alterations, considered among the physiological resistance mechanisms. Two groups which are Ae. albopictus samples that were collected from the city of Aydın in Turkey (study group) and insecticide-free and laboratory-grown Ae. albopictus samples (control group) were the subjects of the study. The acetylcholinesterase (AChE), esterase, oxidase, and glutathione-s-transferase (GST) enzyme levels and specific activity assays were carried out in both groups. All enzyme analyses were conducted according to World Health Organization’s protocol, and protein determination was done by the Bradford method. In conclusion, the two groups had significant differences in all enzyme levels. This study, which revealed such enzyme differences in insecticide-resistant Aedes albopictus, might guide future genetic studies that will explain these enzyme differences and new studies for selecting insecticides that will not harm the environment and humans. [ABSTRACT FROM AUTHOR]

A cataluminescence (CTL) method has been developed for the rapid determination of acetic acid in enzyme products. The NiMn LDH/CNT/GO was synthesized based on the nanohybridization of NiMn layered double hydroxide (NiMn LDH), carbon nanotubes (CNTs), and graphene oxide (GO). The composite has excellent CTL activity against acetic acid. It could be ascribed to the larger specific surface area and more exposure to active sites. NiMn LDH/CNT/GO is used as a catalyst in the CTL method based on its special structure and advantages. There is a linear relationship between CTL response and the acetic acid concentration in the range 0.31–12.00 mg·L−1 with the detection limit of 0.10 mg·L−1. The developed method is rapid and takes only about 13 s. The method is applied to the determination of acetic acid in enzyme samples with little sample preparation. The result of the CTL method shows good agreement with that of the gas chromatography method. The proposed CTL method possesses promising potential in the quality monitoring of enzymes. [ABSTRACT FROM AUTHOR]

Objective: A better understanding of mechanisms regulating lipogenesis and adipogenesis is needed to overcome the obesity pandemic. We aimed to study the relationship of the transcript levels of peroxisome proliferator activator receptor γ (PPARγ), CCAAT/enhancer-binding protein alpha (C/EBP-α), liver X receptor (LXR), sterol regulatory element-binding protein-1c (SREBP-1c), fatty acid synthase (FAS), and acetyl-CoA carboxylase (ACC) in subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) from obese and normal-weight women with a variety of anthropometric indices, metabolic and biochemical parameters, and insulin resistance. Methods: Real‐time PCR was done to evaluate the transcript levels of the above‐mentioned genes in VAT and SAT from all participants. Results: Using principal component analysis (PCA) results, two significant principal components were identified for adipogenic and lipogenic genes in SAT (SPC1 and SPC2) and VAT (VPC1 and VPC2). SPC1 was characterized by relatively high transcript levels of SREBP1c, PPARγ, FAS, and ACC. However, the second pattern (SPC2) was associated with C/EBPα and LXR α mRNA expression. VPC1 was characterized by transcript levels of SREBP1c, FAS, and ACC. However, the VPC2 was characterized by transcript levels of C/EBPα, LXR α, and PPARγ. Pearson's correlation analysis showed that unlike SPC2, which disclosed an inverse correlation with body mass index, waist and hip circumference, waist to height ratio, visceral adiposity index, HOMA-IR, conicity index, lipid accumulation product, and weight-adjusted waist index, the VPC1 was positively correlated with above-mentioned obesity indices. Conclusion: This study provided valuable data on multiple patterns for adipogenic and lipogenic genes in adipose tissues in association with a variety of anthropometric indices in obese subjects predicting adipose tissue dysfunction and lipid accumulation. [ABSTRACT FROM AUTHOR]

Objective: Statin-associated autoimmune myopathy (SAAM) is a rare adverse event characterized by progressive muscle symptoms despite discontinuation, requiring immunosuppressive therapy for remission. The objective of this review was to characterize SAAM, for timely detection, while examining the literature for effective treatment considerations. Methods: PubMed search was conducted from 2010 to 2020 was for relevant case series and studies of at least 8 patients displaying muscle discomfort or weakness, anti-HMGCR antibodies, exposure to statins, and biopsies consistent with SAAM. Results: Three case series and 3 case cohort studies identified 199 patients with SAAM. Exhibiting a mean age of 63.74 years, patients were more likely Caucasian (81%) and female (1.2X), and required a mean duration of 4.75 years before symptomatic. The presentation involved proximal muscle weakness (94%), myalgias (37%) and dysphagia (23%), accompanied by a mean creatinine kinase of 6383 IU/L. Most patients (57%) required 2 or more immunosuppressive (IMS) agents to achieve 62% remission. After 2 years of treatment, 15% of patients without remission reported symptomatic improvement, while another 12% were refractory to treatment. Conclusion: Delayed onset of SAAM greater than 4 years from statin initiation may create a low index of suspicion. However, progression of symptoms beyond 2 months from statin discontinuation and positive anti-HMGCR antibodies requires immunosuppressive agents. Data and expert opinion support use of at least two IMS medications upon diagnosis for a minimum of 2 years. Therapy success depends on timely recognition and initiation of IMS combinations to achieve earlier remission and symptomatic improvement. [ABSTRACT FROM AUTHOR]

Despite remarkable initial responses of anaplastic lymphoma kinase (ALK) inhibitors in ALK-positive non-small cell lung cancer (NSCLC) patients, cancers eventually develop resistance within one to two years. This study aimed to compare the properties of iruplinalkib (WX‑0593) with other ALK inhibitors and report the comprehensive characterization of iruplinalkib against the crizotinib resistance. The inhibitory effect of iruplinalkib on kinase activity was detected. A kinase screen was performed to evaluate the selectivity of iruplinalkib. The effect of iruplinalkib on related signal transduction pathways of ALK and c-ros oncogene 1 (ROS1) kinases was examined. The cellular and in vivo activities of ALK inhibitors were compared in engineered cancer-derived cell lines and in mice xenograft models, respectively. Human hepatocytes derived from three donors were used for evaluating hepatic enzyme inducing activity. HEK293 cell lines expressing transportors were used to invesigated the drug interaction potential mediated by several transporters. The results showed iruplinalkib potently inhibited the tyrosine autophosphorylation of wild-type ALK, ALKL1196M, ALKC1156Y and epidermal growth factor receptor (EGFR)L858R/T790M. The inhibitory effects of iruplinalkib in patient-derived xenograft and cell line-derived xenograft models were observed. Moreover, iruplinalkib showed robust antitumor effects in BALB/c nude mice xenograft models with ALK-/ROS1-positive tumors implanted subcutaneously, and the tumor suppressive effects in crizotinib-resistant model was significantly better than that of brigatinib. Iruplinalkib did not induce CYP1A2, CYP2B6 and CYP3A4 at therapeutic concentration, and was also a strong inhibitor of MATE1 and MATE2K transporters, as well as P-gp and BCRP. In conclusion, iruplinalkib, a highly active and selective ALK/ROS1 inhibitor, exhibited strong antitumor effects in vitro and in crizotinib-resistant models. [ABSTRACT FROM AUTHOR]

Fufang Zhenzhu Tiaozhi (FTZ) capsule is a hospital preparation of a patented traditional Chinese medicine compound. FTZ has been clinically used for nearly 13 years in the treatment of diabetes and glycolipid metabolic diseases. With the significant benefits of SGLT2 inhibitor in patients with diabetic kidney disease (DKD), it provides a research avenue to explore the mechanism of FTZ in treating this disease based on glycolysis pathway. To explore the pharmacological characteristics of FTZ in DKD mice and its impact on the glycolysis pathway. We induced a DKD model in C57BL/6 mice by injection of streptozotocin (STZ) combined with long-term high-fat diet. We administered three doses of FTZ for 12 weeks of treatment. Kidney function, blood lipid levels, glucose tolerance, and key glycolytic enzymes were evaluated. Renal pathological changes were observed using HE, MASSON, and PAS staining. The potential targets of the active ingredients of FTZ in the glycolysis pathway were predicted using network pharmacology and molecular docking. Validation was performed using immunohistochemistry and Western blotting. FTZ effectively reduces blood glucose, total cholesterol, triglyceride, low density lipoprotein cholesterol, 24 h proteinuria, serum creatinine, blood urea nitrogen, and increases urinary glucose levels. Glucose tolerance and renal pathological changes were significantly improved by FTZ treatment. Pinusolidic acid, a component of FTZ, shows good binding affinity with three active pockets of SGLT2. WB and immunohistochemistry revealed that FTZ significantly inhibits the expression of SGLT2 and its glycolytic related proteins (GLUT2/PKM2/HK2). Hexokinase, pyruvate kinase, and lactate dehydrogenase in the kidney were also significantly inhibited by FTZ in a dose-dependent manner. FTZ may alleviate the progression of DKD by inhibiting the activation of the SGLT2/glycolytic pathway. Our study provides new insights into the clinical application of FTZ in DKD. FTZ reduces blood glucose and lipid levels, improves kidney function and glucose tolerance by inhibiting SGLT2 and the glycolytic pathway, potentially helping to alleviate diabetic kidney disease (DKD). One of the components of FTZ, pinusolidic acid, has good binding affinity with three active sites of SGLT2. FTZ significantly inhibits SGLT2 and its related glycolysis protein expression, and dose-dependently inhibits hexokinase, pyruvate kinase, and lactate dehydrogenase in the kidneys. The study concludes that FTZ may help to alleviate the progression of DKD by inhibiting the SGLT2/glycolysis pathway, providing new insights into the clinical application of FTZ in DKD. Abbreviation: SGLT2, Sodium-glucose co-transporter 2; FTZ, Fufang Zhenzhu Tiaozhi capsule; DKD, Diabetic kidney disease; STZ, Streptozotocin; LDL-C, Low density lipoprotein cholesterol; HDL-C, High density lipoprotein cholesterol; TC, Total cholesterol; TG, Triglyceride; UACR, Urinary albumin creatinine ratio; Src, Serum creatinine; BUN, Blood urea nitrogen; PK, Pyruvate kinase; HK, Hexokinase; HK2, Hexokinase II; PKM2, pyruvate kinase M2; GLUT2, Glucose transporter-2. [Display omitted] • FTZ promotes urinary glucose excretion and improves abnormal renal glycolysis. • Bioinformatics analysis has revealed the potential connection between the active components of FTZ and SGLT2. • FTZ ameliorates renal injury in mice with diabetic kidney disease via the SGLT2/glycolysis pathway. [ABSTRACT FROM AUTHOR]

Simple Summary: Cancer cells exhibit unique metabolic properties, including a high requirement for methionine. However, the mechanism that explains how cancer cells suffer from the absence of methionine more significantly than healthy cells remains elusive. Methionine is essential for epigenetic reprogramming of cells, both at the DNA level and for post-translation histone code modification. The post-translational modifications of histone variants (hPTMs) in normal and cancer cells were characterized by mass-spectrometry in the absence and presence of methionine gamma-lyase (MGL), a bacterial enzyme that degrades methionine and inhibits the growth of cancer cells. Results indicate a complex pattern of PTMs on histone variants and striking differences between normal and cancer cells that might help in the understanding of the molecular mechanisms triggered by methionine depletion and in the fine-tuning of MGL-based cancer therapy. Methionine is an essential amino acid involved in the formation of polyamines and a precursor metabolite for DNA and protein methylation. The dependence of cancer cells on methionine has triggered extensive investigations aimed at its targeting for cancer therapy, including the exploitation as a therapeutic tool of methionine γ-lyase (MGL), a bacterial enzyme that degrades methionine, capable of inhibiting cancer cells growth due to methionine starvation. We have exploited the high-resolution power of mass spectrometry to compare the effects of reduced availability of the methyl donor SAM, induced by MGL treatment, on the post-translational modifications of the histone tails in normal Hs27 and cancer HT-29 cells. In the absence of MGL, our analysis detected a three-fold higher relative abundance of trimethylated K25 of H1.4 in HT-29 than Hs27 cells, and a complex pattern of methylated, unmethylated and acetylated peptides in H2 and H3.3. In the presence of MGL, in HT-29, the peptide H2A1_4_11 is predominantly unmodified with mono-methylated K5 increasing upon treatment, whereas in Hs27 cells, H2A1_4_11 is monomethylated at K5 and K9 with these marks decreasing upon treatment. The time dependence of the effects of MGL-mediated methionine depletion on PTMs of histone variants in HT-29 cancer cells was also monitored. Overall, our present data on histone variants H1, H2A, H2B as well as H3.3 integrated with our previous studies on histones H3 and H4, shed light on the epigenetic modifications associated with methionine starvation and associated cancer cell death. [ABSTRACT FROM AUTHOR]

Summary: The effects of polymorphisms in CYP3A4 (20230G > A), CYP3A5 (6986A > G), ABCB1 (1236C > T, 2677G > T/A, 3435C > T), ABCG2 (421C > A), and ABCC2 (-24C > T) on the area under the concentration–time curve (AUC) of osimertinib in 23 patients with non-small cell lung cancer were investigated. Blood sampling was performed just prior to and at 1, 2, 4, 6, 8, 12, and 24 h after osimertinib administration at the steady-state on day 15 after beginning therapy. The osimertinib AUC0-24 was significantly correlated with age (P = 0.038), serum albumin (P = 0.002), and serum creatinine (P = 0.012). Additionally, there were significant differences in the AUC0-24 of osimertinib among the groups administered vonoprazan, histamine 2-receptor antagonists or esomeprazole, and no acid suppressants (P = 0.021). By contrast, there were no significant differences in the AUC0-24 of osimertinib between genotypes of CYP3A4/5 or ABC transporters. Furthermore, there were no significant differences in the AUC0-24 of osimertinib between patients with diarrhea, skin rash, or hepatotoxicity and those without these conditions. In multivariate analysis, only serum albumin value was an independent factor predicting the AUC0-24 of osimertinib. Analysis of CYP3A4/5 and ABC transporter polymorphisms before osimertinib therapy may not predict the efficacy or side effects of osimertinib. The lower serum albumin values were associated with an increase in the AUC0-24 of osimertinib; however, further studies are needed to assess the factors contributing to the interindividual variability of osimertinib pharmacokinetics. [ABSTRACT FROM AUTHOR]

Objective: Neuron-specific enolase is an established biomarker of neuronal damage. This study aimed to reveal the relationship between serum neuron-specific enolase level and con)tinuous interictal discharges in a group of encephalopathy with electrical status epilepticus in sleep patients for the first time and determine whether there is a neuronal cell loss or damage. Materials and Methods: We analyzed serum neuron-specific enolase levels in patients with an electrical status epilepticus in sleep pattern on their electroencephalographs with age- and sex-matched control subjects. Patients with a spike–wave index of at least 50% and acquired neuropsychological regression were included in the study. Magnetic resonance imaging of all electrical status epilepticus in sleep patients and control subjects included in the study was within normal limits. Neuron-specific enolase is measured by the enzyme-linked immunosorbent assay kit based on the sandwich technique. Results: In this study, 14 patients diagnosed with electrical status epilepticus in sleep and 21 healthy controls were included. The median age of electrical status epilepticus in sleep patients was 7.1 years (min-max: 4.5-10.7 years) and 7.7 years (min-max: 3.2-14 years) in the control subjects. According to the results of serum neuron-specific enolase measurements, the mean ± standard deviation level of neuron-specific enolase was 7.61 ± 3.19 ng/dL for the electrical status epilepticus in sleep group and 6.93 ± 2.55 ng/dL for the control group. Serum neuron-specific enolase levels between electrical status epilepticus in sleep patients and the control group were not statistically significant (P = .749). Conclusion: No significant difference was observed in serum neuron-specific enolase levels between electrical status epilepticus in sleep patients and control subjects. Our results may indicate that frequent interictal discharges do not result in neuronal cell loss or damage in electrical status epilepticus in sleep patients. [ABSTRACT FROM AUTHOR]

Background: The appropriate oxygenation target for mechanical ventilation in comatose survivors of out-of-hospital cardiac arrest is unknown.Methods: In this randomized trial with a 2-by-2 factorial design, we randomly assigned comatose adults with out-of-hospital cardiac arrest in a 1:1 ratio to either a restrictive oxygen target of a partial pressure of arterial oxygen (Pao2) of 9 to 10 kPa (68 to 75 mm Hg) or a liberal oxygen target of a Pao2 of 13 to 14 kPa (98 to 105 mm Hg); patients were also assigned to one of two blood-pressure targets (reported separately). The primary outcome was a composite of death from any cause or hospital discharge with severe disability or coma (Cerebral Performance Category [CPC] of 3 or 4; categories range from 1 to 5, with higher values indicating more severe disability), whichever occurred first within 90 days after randomization. Secondary outcomes were neuron-specific enolase levels at 48 hours, death from any cause, the score on the Montreal Cognitive Assessment (ranging from 0 to 30, with higher scores indicating better cognitive ability), the score on the modified Rankin scale (ranging from 0 to 6, with higher scores indicating greater disability), and the CPC at 90 days.Results: A total of 789 patients underwent randomization. A primary-outcome event occurred in 126 of 394 patients (32.0%) in the restrictive-target group and in 134 of 395 patients (33.9%) in the liberal-target group (hazard ratio, 0.95; 95% confidence interval, 0.75 to 1.21; P = 0.69). At 90 days, death had occurred in 113 patients (28.7%) in the restrictive-target group and in 123 (31.1%) in the liberal-target group. On the CPC, the median category was 1 in the two groups; on the modified Rankin scale, the median score was 2 in the restrictive-target group and 1 in the liberal-target group; and on the Montreal Cognitive Assessment, the median score was 27 in the two groups. At 48 hours, the median neuron-specific enolase level was 17 μg per liter in the restrictive-target group and 18 μg per liter in the liberal-target group. The incidence of adverse events was similar in the two groups.Conclusions: Targeting of a restrictive or liberal oxygenation strategy in comatose patients after resuscitation for cardiac arrest resulted in a similar incidence of death or severe disability or coma. (Funded by the Novo Nordisk Foundation; BOX ClinicalTrials.gov number, NCT03141099.). [ABSTRACT FROM AUTHOR]

Background: Evidence to support the choice of blood-pressure targets for the treatment of comatose survivors of out-of-hospital cardiac arrest who are receiving intensive care is limited.Methods: In a double-blind, randomized trial with a 2-by-2 factorial design, we evaluated a mean arterial blood-pressure target of 63 mm Hg as compared with 77 mm Hg in comatose adults who had been resuscitated after an out-of-hospital cardiac arrest of presumed cardiac cause; patients were also assigned to one of two oxygen targets (reported separately). The primary outcome was a composite of death from any cause or hospital discharge with a Cerebral Performance Category (CPC) of 3 or 4 within 90 days (range, 0 to 5, with higher categories indicating more severe disability; a category of 3 or 4 indicates severe disability or coma). Secondary outcomes included neuron-specific enolase levels at 48 hours, death from any cause, scores on the Montreal Cognitive Assessment (range, 0 to 30, with higher scores indicating better cognitive ability) and the modified Rankin scale (range, 0 to 6, with higher scores indicating greater disability) at 3 months, and the CPC at 3 months.Results: A total of 789 patients were included in the analysis (393 in the high-target group and 396 in the low-target group). A primary-outcome event occurred in 133 patients (34%) in the high-target group and in 127 patients (32%) in the low-target group (hazard ratio, 1.08; 95% confidence interval [CI], 0.84 to 1.37; P = 0.56). At 90 days, 122 patients (31%) in the high-target group and 114 patients (29%) in the low-target group had died (hazard ratio, 1.13; 95% CI, 0.88 to 1.46). The median CPC was 1 (interquartile range, 1 to 5) in both the high-target group and the low-target group; the corresponding median modified Rankin scale scores were 1 (interquartile range, 0 to 6) and 1 (interquartile range, 0 to 6), and the corresponding median Montreal Cognitive Assessment scores were 27 (interquartile range, 24 to 29) and 26 (interquartile range, 24 to 29). The median neuron-specific enolase level at 48 hours was also similar in the two groups. The percentages of patients with adverse events did not differ significantly between the groups.Conclusions: Targeting a mean arterial blood pressure of 77 mm Hg or 63 mm Hg in patients who had been resuscitated from cardiac arrest did not result in significantly different percentages of patients dying or having severe disability or coma. (Funded by the Novo Nordisk Foundation; BOX ClinicalTrials.gov number, NCT03141099.). [ABSTRACT FROM AUTHOR]

Objective: To analyze the expression and clinical significance of E3 ubiquitin ligase (HECW2) in gastric adenocarcinoma using a bioinformatics approach to provide new clues for finding diagnostic and prognostic biomarkers for gastric adenocarcinoma. Methods: The expression of HECW2 in pan-cancer and its relationship with the prognosis of pan-cancer were analyzed by R language. The expression of HECW2 in gastric adenocarcinoma tissues and its relationship with clinicopathological features were analyzed with the help of UCSC Xena, HPA, and Kaplan-Meier Plotter databases. The WB method was used to detect HECW2 protein levels in Chinese gastric adenocarcinoma tissues and their paraneoplastic tissues to verify the analysis results in the database. The relationship between HECW2 and immune infiltration of gastric adenocarcinoma was analyzed with the help of TIMER and Cibersort databases. GO functional analysis and KEGG signaling pathway enrichment analysis and correlation gene analysis were performed for HECW2 in gastric adenocarcinoma by LinkedOmics database. Results: The results of raw signal analysis showed that HECW2 was significantly highly expressed in 12 tumors including gastric adenocarcinoma among 33 different types of tumors (all P<0.05), and the results of WB method showed that HECW2 was also significantly highly expressed in Chinese gastric adenocarcinoma tissues (P<0.05). Higher HECW2 expression level was associated with shorter OS and higher abundance of infiltration of immune cells including CD4+ T cells, macrophages, neutrophils and dendritic cells in gastric adenocarcinoma patients (all P<0.01). In addition, HECW2-associated signaling pathways were mainly enriched in extracellular matrix receptor interactions, adherent spots, cell adhesion and oxidative phosphorylation (all P<0.01). Conclusion: HECW2 is significantly overexpressed in gastric adenocarcinoma tissues and is closely related to the poor prognosis and immune invasion of gastric adenocarcinoma, and has great potential as a prognostic biomarker and target therapy for gastric adenocarcinoma. [ABSTRACT FROM AUTHOR]

Cultivating productive paddy crops on salty soil to maximise production is a challenging approach to meeting the world's growing food demand. Thus, determining salinity tolerance rates in specific paddy cultivars is urgently needed. In this study, the salt tolerance traits of selected paddy cultivars, ADT45 and ADT39, were investigated by analysing germination, metabolites (pigments and biomolecules), and enzymatic (Superoxide dismutase (SOD), Catalase (CAT), and Peroxidase (POD) adaptation strategies as salt-stress tolerance mechanisms. This study found that salinity-induced reactive oxygen species (ROS) were efficiently detoxified by the antioxidant enzymes Superoxide dismutase (SOD), Catalase (CAT), and Peroxidase (POD) in ADT45 paddy varieties, followed by ADT39. Salinity stress had a significant impact on pigments and essential biomolecules in ADT45 and ADT39 paddy cultivars, including total chlorophyll, anthocyanin, carotenoids, ascorbic acid, hydrogen peroxide (H 2 O 2), malondialdehyde, and proline. ADT45 demonstrated a significant relationship between H 2 O 2 and antioxidant enzyme levels, followed by ADT39 paddy but not IR64. Morphological, physiological, and biochemical analyses revealed that ADT45, followed by ADT39, is a potential salt-tolerant rice cultivar. • ADT45 and ADT39 paddy varieties showed fine tolerance to salt stress. • Salinity-induced ROS were efficiently detoxified by the antioxidant enzymes. • Salinity stress influenced pigments & essential biomolecules in ADT45 & ADT39. • ADT45 showed a significant correlation between H 2 O 2 and antioxidant enzyme levels. [ABSTRACT FROM AUTHOR]

Fluorescent proteins (FPs) based sensing strategies have been widely developed in bioanalytical applications. However, it still remains a challenge to realize the efficient and precise analysis. Recently, FPs chromophore and its derivatives with favorable optical properties are well-designed to simulate and replace the function of FPs by specifically binding to the selected fluorescent light-up aptamers (FLAPs), it is worthy note that there is still a lack of systematic approaches to remold these FPs mimics with enzyme-responsive functions. Taking alkaline phosphatase (ALP) as a research model, we here propose an ALP-responsive fluorescent DNA aptasensor (p-DAA) by utilizing a truncated Lettuce (tLet-10) FLAP and a phosphorylated DFHBI-derivative (ALP-DFSS) for achieving ALP activity analysis. Taking the HBI-chromophore as the remolding molecular template, ALP-DFSS has been successfully synthesized and characterized to response to ALP activity. With phosphate group substitution in ALP-DFSS, the decayed fluorescence of p-DAA can be observed by forming the complex of tLet-10/ALP-DFSS. While the turn-on fluorescence of p-DAA will be obtained by the ALP-triggered dephosphorylation process and further exhibit the label-free, fast-response, specific, and sensitive detection performances for both evaluating ALP activity and the inhibition efficiency of ALP inhibitor. Moreover, p-DAA allow the spiked ALP analysis in serum samples and is also applied for realizing the visual differentiation between normal cell line and cancer cell line by responding the endogenous ALP, which holds the great potential in guiding the chromophores functionalization and exploring aptasensing related biological research. [Display omitted] • Achieving the enzyme-responsive function chromophores derivatives. • Constructing the ALP-responsive fluorescent DNA aptasensor. • Achieving the facile, label-free, fast-response, and visual detection performances. • Realizing the ALP detection and imaging in biological samples. [ABSTRACT FROM AUTHOR]

Kudzu-resistant starch was prepared by pullulanase debranching recrystallization. Based on single factor experiments, the response surface optimization method was used to analyze the optimal conditions for the preparation of kudzu-resistant starch. The results showed that the highest yield of 90.46% was obtained at the enzymatic reaction time of 4 h, temperature of 56°C, and amount of enzyme being 57 U/g. After debranching, the content of the resistant starch was higher than native starch. The crystal structure of the debranched starch changed from A-type to a mixture of B+V-type. The results of scanning electron microscope showed that the surface of the native starch granule was smooth and varied in size, showing a spherical or irregular polyhedral shape, while the debranched starch granule was irregular in shape and size and the surface was rough and gully. The granule morphology of the modified starch granule was also changed. [ABSTRACT FROM AUTHOR]

Objective To develop and validate a prognostic model for LEF discontinuation with abnormal blood test results. Methods Data from the Clinical Practice Research Datalink Gold and Aurum were used for model development and external validation, respectively. Participants prescribed LEF between 1 January 2007 and 31 December 2019 were followed up from 6 months after the first general practitioner prescription to the earliest of date of outcome, death, 5 year follow-up or 31 December 2019. Candidate prognostic factors were ascertained using theory and data-driven approaches. Penalized Cox regression was performed to develop the risk equation, followed by internal validation using 500 bootstraps to correct for optimism. Multiple imputation was applied to handle missing data. Model performance was assessed in terms of calibration and discrimination. Results Data for 1487 and 2329 participants contributing 3140 and 5246 person-years follow-up were included in the development and validation cohorts, respectively. Thirteen candidate predictors were included in the model. Epilepsy and either cytopenia or elevated liver enzymes during the first 6 months of shared-care LEF prescription were strong predictors of drug discontinuation with a hazard ratio of 4.39 (95% CI 1.74, 11.06) and 3.06 (2.15, 4.35), respectively. The unadjusted and optimism-adjusted calibration slope in development data was 1.00 (95% CI 0.75, 1.25) and 0.72 (95% CI 0.47, 0.97), respectively. The calibration slope in validation data was 0.91 (95% CI 0.74, 1.07). The model showed prognostic separation with an optimism-adjusted Royston D statistic of 0.73 (95% CI 0.44, 1.02). Conclusion We have developed and externally validated an easy-to-use prognostic model that may be used to risk stratify monitoring for LEF toxicity and to make informed choices about risks when choosing treatments. [ABSTRACT FROM AUTHOR]

• Unrelated patients with suspected familial hypercholesterolemia • Monogenic etiology of familial hypercholesterolemia in 25.6% adults and 77% children • LDL-cholesterol as a major parameter to suspect monogenic hypercholesterolemia Familial hypercholesterolemia (FH) is the most common genetic disorder associated with a high risk for premature atherosclerotic cardiovascular disease attributable to increased levels of LDL-cholesterol (LDL-C) from birth. FH is both underdiagnosed and undertreated. We describe the clinical, biological, and genetic characteristics of 147 patients in France with clinical FH (including a group of 26 subjects aged < 20 years); we explore how best to detect patients with monogenic FH. We retrospectively reviewed all available data on patients undergoing genetic tests for FH from 2009 to 2019. FH diagnoses were based on the Dutch Lipid Clinics Network (DLCN) scores of adults, and elevated LDL-C levels in subjects < 20 years of age. We evaluated LDLR, APOB , and PCSK9 status. The mutations of adults (in 25.6% of all adults) were associated with DLCN scores indicating "possible FH," "probable FH, and "definitive FH" at rates of 4%, 16%, and 53%, respectively. The areas under the ROC curves of the DLCN score and the maximum LDL-C level did not differ (p = 0.32). We found that the pediatric group evidenced more monogenic etiologies (77%, increasing to 91% when an elevated LDL-C level was combined with a family history of hypercholesterolemia and/or premature coronary artery disease). Diagnosis of monogenic FH may be optimized by screening children in terms of their LDL-C levels, associated with reverse-cascade screening of relatives when the children serve as index cases. [ABSTRACT FROM AUTHOR]

Background: Dermatophytes are keratinophilic fungi that affect the stratum corneum of the skin and keratinous structures. Violent factors play a vital role in the pathogenesis and antifungal resistance of dermatophytes. Objective: This study aims to evaluate the activity of extracellular enzymatic and biofilm formation as virulence factors of dermatophyte isolates. Methods: Fifty-eight dermatophyte isolates belonged to 27 Trichophyton. rubrum (46.6%), 19 T. mentagrophytes (32.8%), and 12 Microsporum. canis (20.7%) for evaluating the activity of phospholipase, hemolysin, proteinase, and biofilm formation were examined. The biofilm formed was analyzed by scanning electron microscopy (SEM). Findings: Evaluation of extracellular enzymes production revealed that 86.2%, 77.6 %, and 57% of dermatophyte strains were shown to be phospholipase, hemolysin, and proteinase producers, respectively. Furthermore, all isolates of T. rubrum and M. canis can produce phospholipase and hemolysin, respectively. There was a statistically significant difference between phospholipase activity and dermatophyte strains (P<0.05). In addition, biofilm formation ability was observed in 41.5% of dermatophyte isolates. The highest level of biofilm production was found in 93% of dermatophytes isolated from nail chips. A significant difference between biofilm formation with dermatophyte isolates and different body sites was observed (P <0.05). Conclusion: The activity of hydrolytic enzymes and biofilm formation as important pathogenic factors may play a role in the persistence of dermatophytosis infections. Our results showed that dermatophyte isolates have enzymatic activity and biofilm production at different levels. Therefore, understanding the function of these factors is essential to controlling the spread of dermatophytosis infection. [ABSTRACT FROM AUTHOR]

Bile acid is a derivative of cholinergic acid (steroidal parent nucleus) that plays an important role in digestion, absorption, and metabolism. In recent years, bile acids have been identified as signaling molecules that regulate self-metabolism, lipid metabolism, energy balance, and glucose metabolism. The detection of fine changes in bile acids caused by metabolism, disease, or individual differences has become a research hotspot. At present, there are many related techniques, such as enzyme analysis, immunoassays, and chromatography, that are used for bile acid detection. These methods have been applied in clinical practice and laboratory research to varying degrees. However, mainstream detection technology is constantly updated and replaced with the passage of time, proffering new detection technologies. Previously, gas chromatography (GS) and gas chromatography-mass spectrometry (GC-MS) were the most commonly used for bile acid detection. In recent years, high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has developed rapidly and has gradually become the mainstream bile acid sample separation and detection technology. In this review, the basic principles, development and progress of technology, applicability, advantages, and disadvantages of various detection techniques are discussed and the changes in bile acids caused by related diseases are summarized. [ABSTRACT FROM AUTHOR]

Background: Serum creatinine concentration is a primary component of Bedside Schwartz equation for estimated glomerular filtration rate (eGFR) in children. To standardize creatinine measurement, most manufacturers have adopted calibration procedures traceable to isotope dilution mass spectrometry (IDMS) using National Institute of Standards and Technology reference material. However, reference material representing much lower creatinine concentrations seen in children is not available and it is unclear how well commercial assays perform at pediatric levels. Methods: One thousand nine hundred seventy-one specimens from consecutive children <19 years, with creatinine ≤0.8 mg/dL by Abbott Jaffe method were included. Creatinine measurements were compared between Abbott-Jaffe and Abbott-enzymatic methods. Furthermore, we evaluated performance of six commercial creatinine assays at concentrations seen in pediatric patients utilizing IDMS traceable serum samples. Results: Median difference (enzymatic-Jaffe) for prepubertal females was –0.18 mg/dL (2.5%tile, 97.5%tile: –0.30, –0.06), –0.12 mg/dL (–0.25, –0.00) for pubertal females, –0.17 mg/dL (–0.30, –0.04) for prepubertal males, –0.11 mg/dL (–0.24, 0.01) for pubertal males. Bias appeared proportional for each subgroup and decreased as creatinine concentrations increased. Using IDMS traceable samples, the greatest inter-assay variability was seen with the lowest creatinine levels (target 0.273 mg/dL), where 67% (4/6) of methods failed to reach minimal bias specification of 8% (range –7.5 to 86%). For samples with higher creatinine targets (0.440–0.634 mg/dL), two methods failed to meet minimal bias specification, whereas four showed bias <8%. Conclusion: Many commonly used creatinine assays remain inaccurate for pediatric populations after over a decade of nationwide efforts to standardize measurements. When creatinine-based eGFR is used for chronic kidney disease (CKD) staging in children, large inter-assay variability can lead to disease misclassification, inappropriate diagnostic and therapeutic interventions. A higher resolution version of the Graphical abstract is available as Supplementary information [ABSTRACT FROM AUTHOR]

We disclose a mild and practical catalyst-free transformation for the expeditious construction of biuret-guanidine derivatives using aromatic isocyanates. This synthetic transformation is featured with mild reaction conditions and high efficiency. [ABSTRACT FROM AUTHOR]

Bile acid is a derivative of cholinergic acid (steroidal parent nucleus) that plays an important role in digestion, absorption, and metabolism. In recent years, bile acids have been identified as signaling molecules that regulate self-metabolism, lipid metabolism, energy balance, and glucose metabolism. The detection of fine changes in bile acids caused by metabolism, disease, or individual differences has become a research hotspot. At present, there are many related techniques, such as enzyme analysis, immunoassays, and chromatography, that are used for bile acid detection. These methods have been applied in clinical practice and laboratory research to varying degrees. However, mainstream detection technology is constantly updated and replaced with the passage of time, proffering new detection technologies. Previously, gas chromatography (GS) and gas chromatography-mass spectrometry (GC-MS) were the most commonly used for bile acid detection. In recent years, high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has developed rapidly and has gradually become the mainstream bile acid sample separation and detection technology. In this review, the basic principles, development and progress of technology, applicability, advantages, and disadvantages of various detection techniques are discussed and the changes in bile acids caused by related diseases are summarized.

Enzymes control chemical reactions that are key to life processes, and allow them to take place on the time scale needed for synchronization between the relevant reaction cycles. In addition to general interest in their biological roles, these proteins present a fundamental scientific puzzle, since the origin of their tremendous catalytic power is still unclear. While many different hypotheses have been put forward to rationalize this, one of the proposals that has become particularly popular in recent years is the idea that dynamical effects contribute to catalysis. Here, we present a critical review of the dynamical idea, considering all reasonable definitions of what does and does not qualify as a dynamical effect. We demonstrate that no dynamical effect (according to these definitions) has ever been experimentally shown to contribute to catalysis. Furthermore, the existence of non-negligible dynamical contributions to catalysis is not supported by consistent theoretical studies. Our review is aimed, in part, at readers with a background in chemical physics and biophysics, and illustrates that despite a substantial body of experimental effort, there has not yet been any study that consistently established a connection between an enzyme's conformational dynamics and a significant increase in the catalytic contribution of the chemical step. We also make the point that the dynamical proposal is not a semantic issue but a well-defined scientific hypothesis with well-defined conclusions. [ABSTRACT FROM AUTHOR]

The human transmembrane protease serine 2 (TMPRSS2) protein plays an important role in prostate cancer progression. It also facilitates viral entry into target cells by proteolytically cleaving and activating the S protein of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In the current study, we used different available tools like SIFT, PolyPhen2.0, PROVEAN, SNAP2, PMut, MutPred2, I-Mutant Suite, MUpro, iStable, ConSurf, ModPred, SwissModel, PROCHECK, Verify3D, and TM-align to identify the most deleterious variants and to explore possible effects on the TMPRSS2 stability, structure, and function. The six missense variants tested were evaluated to have deleterious effects on the protein by SIFT, PolyPhen2.0, PROVEAN, SNAP2, and PMut. Additionally, V160M, G181R, R240C, P335L, G432A, and D435Y variants showed a decrease in stability by at least 2 servers; G181R, G432A, and D435Y are highly conserved and identified posttranslational modifications sites (PTMs) for proteolytic cleavage and ADP-ribosylation using ConSurf and ModPred servers. The 3D structure of TMPRSS2 native and mutants was generated using 7 meq as a template from the SwissModeller group, refined by ModRefiner, and validated using the Ramachandran plot. Hence, this paper can be advantageous to understand the association between these missense variants rs12329760, rs781089181, rs762108701, rs1185182900, rs570454392, and rs867186402 and susceptibility to SARS-CoV-2. [ABSTRACT FROM AUTHOR]

Background: The prevalence of abnormal cardiovascular magnetic resonance (CMR) findings in recovered coronavirus disease 2019 (COVID-19) patients is unclear. This study aimed to investigate the prevalence of abnormal CMR findings in recovered COVID-19 patients. Methods: A systematic literature search was performed to identify studies that report the prevalence of abnormal CMR findings in recovered COVID-19 patients. The number of patients with abnormal CMR findings and diagnosis of myocarditis on CMR (based on the Lake Louise criteria) and each abnormal CMR parameter were extracted. Subgroup analyses were performed according to patient characteristics (athletes vs. non-athletes and normal vs. undetermined cardiac enzyme levels). The pooled prevalence and 95% confidence interval (CI) of each CMR finding were calculated. Study heterogeneity was assessed, and meta-regression analysis was performed to investigate factors associated with heterogeneity. Results: In total, 890 patients from 16 studies were included in the analysis. The pooled prevalence of one or more abnormal CMR findings in recovered COVID-19 patients was 46.4% (95% CI 43.2%–49.7%). The pooled prevalence of myocarditis and late gadolinium enhancement (LGE) was 14.0% (95% CI 11.6%–16.8%) and 20.5% (95% CI 17.7%–23.6%), respectively. Further, heterogeneity was observed (I2 > 50%, p < 0.1). In the subgroup analysis, the pooled prevalence of abnormal CMR findings and myocarditis was higher in non-athletes than in athletes (62.5% vs. 17.1% and 23.9% vs. 2.5%, respectively). Similarly, the pooled prevalence of abnormal CMR findings and LGE was higher in the undetermined than in the normal cardiac enzyme level subgroup (59.4% vs. 35.9% and 45.5% vs. 8.3%, respectively). Being an athlete was a significant independent factor related to heterogeneity in multivariate meta-regression analysis (p < 0.05). Conclusions: Nearly half of recovered COVID-19 patients exhibited one or more abnormal CMR findings. Athletes and patients with normal cardiac enzyme levels showed a lower prevalence of abnormal CMR findings than non-athletes and patients with undetermined cardiac enzyme levels. Trial registration The study protocol was registered in the PROSPERO database (registration number: CRD42020225234). [ABSTRACT FROM AUTHOR]

Background: In this study, it was aimed to investigate the demographic, etiological and prognostic characteristics of the cases who applied to the emergency department due to intoxication and how much of the intoxication cases the liver was affected. Materials and Methods: The files of the patients who appliedto the emergency service of Harran University Medical Faculty Hospital between January 2018 and September 2019 were retrospectively scanned. Age, gender, anamnesis features and blood tests were examined. Results: 137 of the 211 cases (64.9%) were female, the meanage was 30.3 ± 13.12, the agerange was between 18-72. Of the cases, 108 (51.2%) were drug, 39 (18.5%) were scorpion bites, 23 (10.9%) were food, 14 (6.6%) were insect-mice, 12 (5.7%) were snake bites, 9 (4.3%) were corrosive substances, cannabis-ecstasy in 4 cases and alcohol and carbon monoxide intoxication in 1 case each. Transaminases (ALT, AST) were found to be elevated in 11 patients, cholestasis enzymes (ALP, GGT) or bilirubin in 16 patients. By evaluating the recommendation and clinical conditions of the Poison Counseling Center, 136 (64.5%) cases were followed in the intensive care unit (ICU) and 44 (20.8%) Cases were followed in the service. One of the cases followed up due to drug intoxication died due to organ failure during follow-up in the ICU, and liver enzymes of this patient were high. Conclusions: The most common cause of intoxication applications to the emergency department of ourhospital is medication, followed by scorpion-snake bites and food poisoning. Intoxication is mostly seen in younger ages and in female patients. The liver is affected in about ten percent of patients, elevated liver enzymes may indicate a poor prognosis. [ABSTRACT FROM AUTHOR]

To explore the changes in amplitude-integrated electroencephalography (aEEG), neuron-specific enolase (NSE), and S100B in neonates with brain injury induced by neonatal hyperbilirubinemia (NHB). 67 neonates with brain injury induced by NHB admitted to our hospital from March 2016 to October 2018 were included in a brain injury group (BIG), and 82 neonates with NHB but without brain injury in our hospital during the same period were included in a non-BIG. The two groups were compared regarding the rates of normal and abnormal aEEG results. The proportion of normal aEEG results in the BIG was significantly lower than that in the non-BIG, and the proportion of moderately and severely abnormal aEEG results in the BIG were both significantly higher than those in the non-BIG. The BIG showed significantly higher NSE and S100B levels than those of the non-BIG. The ROC curve for predicting prognosis showed that the AUC of aEEG, NSE, S100B, and the combined detection are 0.780, 0.754, 0.743, 0.788. The AUC > 0.700 indicated a good predictive value for the prognosis. The combination of aEEG, NSE, and S100B has good value in diagnosing injury induced by NHB and can predict prognosis moderately well. [ABSTRACT FROM AUTHOR]

Abstract: Seed vigor may be determinant for field performance, especially under water restriction conditions. Sunflower is a crop subject to these conditions in the field and, therefore, the aim of this study was to evaluate the physiological and antioxidant changes in sunflower seeds under water restriction. Two lots of sunflower seeds (cv. Hélio 253) with different vigor levels were used. After initial characterization, seeds were placed to germinate under water potentials of 0.0, -0.2, -0.4, -0.6 and -0.8 MPa and evaluated by tests of germination, first germination count, shoot length and primary root length. The activities of the antioxidant enzymes SOD, CAT, POX and APX were also evaluated at 0, 2, 4 and 6 days after sowing. Water restriction led to a decrease in germination and slower seedling growth, regardless of seed vigor level. SOD activity was similar in the two lots, with reduction in activity four days after sowing. CAT activity was affected differently during germination in the two lots, and it was generally higher in the most vigorous lot. In higher vigor seeds, there was lower POX activity in water restriction treatments compared to the control. In general, seeds of lower vigor have lower capacity for activation of antioxidant enzymes, especially peroxidases.

Objectives: The aim of this study was to collect consistent data on the efficacy and safety and evaluation hepatotoxicity of intravenous acetaminophen for the treatment of PDA in preterm infants. Methods: This is an observational longitudinal prospective study on 46 preterm infants with PDA who treated with high dose of acetaminophen and evaluated with echocardiography and serum liver enzymes at Hafez and Zeinabiyeh hospitals from January 2016 to December 2019. Result: Forty-six preterm infants with PDA treated with intravenous acetaminophen. Rate of closure of PDA was 82.6. There was no significant difference after treatment regarding AST, ALT, Albumin, total and direct bilirubin (P value > 0.05) and no adverse side effects were observed in association with intravenous acetaminophen. Conclusion: High dose of acetaminophen is not more effective than that with standard doses although without hepatotoxic side effect for PDA closure. [ABSTRACT FROM AUTHOR]

Array‐based analytical platforms have a distinct advantage of being able to detect analytes rapidly and simultaneously with the use of very small quantities of reagents and samples. However, the analyses of enzyme activities using this technique remain challenging, as the heterogeneous interface between the enzyme in the solution and the probe on the array surface impede the efficient binding and the subsequent enzymatic reaction. In this paper, we showed that the combination of affinity‐switchable biotin (ASB) probes and a novel click‐chemistry based immobilization method can overcome this heterogeneity problem. Three ASB probes were synthesized for the detection of alkaline phosphatase, nitroreductase, and the fluoride ion. This approach can be applied to determine the relative levels of phosphatase in different cell lines. We believe that the new strategy holds great potential for the effective sensing of a wide range of biomolecules in the future. [ABSTRACT FROM AUTHOR]