Assay Method Research Articles

Method development and validation of an analytical quality by design ultrafast liquid chromatographic method for the determination of bedaquiline from pharmaceutical bulk and nanoemulsions.

Bedaquiline (BDQ) is a drug used to treat multidrug-resistant tuberculosis (MDR-TB). It exhibits exposure-dependent efficacy in eliminating Mycobacterium tuberculosis (Mtb). An easy, efficient and precise reverse-phase ultrafast liquid chromatography (RP-UFLC) method was developed to validate the free base of the antitubercular medication BDQ. BDQ was separated using a 10:90 v/v mobile phase of ammonium acetate buffer solution (pH = 5.4) and high-performance liquid chromatography-grade methanol, with a flow rate of 1.5mL/min and a UV detection wavelength of 226 nm. By using the Box-Behnken design (BBD) and response surface methodology (RSM), the method was optimised by varying critical analytical attributes (CAA) and critical performance attributes (CPAs) namely ammonium acetate fraction (%), flow rate (ml/min), buffer system molarity (M) and pH. BDQ was eluted at 7.5min utilising isocratic elution. The method was linear in the concentration range of 0.5-300 μg/mL with limit of detection values of 0.039 μg/mL and limit of quantification of 0.12 μg/mL. The results indicate that this validated method can be used as an alternative method for assay of BDQ.

Identification of β-globin gene mutations among transfusion-dependent β-thalassemia patients

Abstract BACKGROUND: β-thalassemias are widely distributed in Mediterranean and Middle Eastern countries, including Iraq. There are more than 400 transfusion-dependent β-thalassemia patients registered in the thalassemia center. β-thalassemia is a significant problem in Karbala as well as other regions of Iraq. The detection of the most frequent mutations is significant to the implementation of an effective preventive program in this area because of the significant burden it places on the local health authorities, patients, and their families. OBJECTIVES: To define the most common mutations and their frequencies among patients with transfusion-dependent β-thalassemia and to evaluate the reverse hybridization strip assay method for the detection of β-thalassemia mutations. PATIENTS, MATERIALS AND METHODS: Sixty transfusion-dependent β-thalassemia patients were recruited from the thalassemia center in Karbala. Blood samples were aspirated from each patient just before blood transfusions for CBC, reticulocyte count, DNA extraction, PCR amplification, and identification of the mutations by reverse hybridization technique using the β-Globin strip assay method. RESULTS: A total of 60 patients with 120 chromosomes were studied, searching for the most common mutations causing β-thalassemia. Among the twelve identified mutations, the six most frequent mutations represented 79.16% of all β-globin defects. These mutations were IVSII-1 (30.83%), IVSI-110 (15.83%), Codon 5 (10.83%), Codon 44 (8.33%), IVSI-1 (6.67%), and IVSI-5 (6.67%). The detection rate of the method used in our population was 96.66%. CONCLUSION: The most frequent mutations encountered were IVSII.1 and IVSI-110, while IVS 2.745 was the least common mutant allele. Reverse hybridization strip assay molecular techniques used in the current study provide an extremely quick, precise, and simple to carry out molecular diagnostic technique for the detection of β-thalassemia mutations.

Standardization of Assay Method of Urease Activity in Inceptisol

Urea is the most important nitrogen fertilizer widely used in crop production and its fate in soils is regulated largely by the soil enzyme urease. The use of precise assay method for urease is an important aspect in regulating activity of soil urease which is the most useful to improve use efficiency of urea, and minimize the problems related to use of urea. In this context, the present laboratory investigation was undertaken with the objective to standardize the assay media components of soil urease activity in Inceptisol of MPKV, Rahuri soils. The Typic Haplustept was selected as a representative of Inceptisol and used for present study of optimizing assay media components for in vivo assay method for measuring activity of urease from Inceptisol. All the assay components viz., substrate concentration (0.2 M), buffer strength (0.05 M), weight of soil (5 g), amount of toluene (0.2 ml) and incubation time (2 h) were found optimum as per Tabatabai and Bremner (1972) except THAM buffer pH. The THAM buffer with a pH 8.5 was found to be optimum for measuring the activity of urease from Inceptisol. The values of urease activity from the 2g weight of soil and 1 h incubation time also found statistically at par with values of 5 g weight of soil and 2 h incubation time, respectively. Hence, the use of 2 g soil and 1 h incubation time in assay procedure are also sufficient for measuring urease activity, if the soil sample size and time are limiting factors.

Association of Activin A Levels and Body Mass Index with Human CGB (Chorionic Gonadotrophin Beta) in Ectopic Pregnancies and Missed Abortion Patients

Background: An ectopic pregnancy occurs when a fertilized egg implants itself—typically in one of the fallopian tubes—outside of the womb. Connecting the ovaries to the womb are tubes called fallopian tubes. An egg lodged in them won't develop into a baby, and if the pregnancy goes on, it could be dangerous for your health. When symptoms do appear, they usually appear between weeks four and twelve of pregnancy. Missed abortion refers to the clinical state in which there is still an intrauterine pregnancy but it is not progressing normally. Only if the diagnosis of an incomplete abortion or an inevitable abortion was ruled out would the gestation be referred to be a missed abortion. Activin A is a protein that participates principally in reproductive functions. In the adult brain, Neuroprotective effects of activin. Pituitary follicle-stimulating hormone (FSH) release is stimulated by activins, which are separated from gonadal fluids and belong to the The transforming growth factor (TGF-beta) superfamily. Activins are disulfide-linked dimeric proteins. Objectives: To estimate the concentrations in serum of Activin A in women who have been diagnosed with (missed abortion) and ectopic pregnancy, to compare them with non- missed abortion (MA) and ectopic pregnancy (Ep) healthy controls, and to explore the impact of human chorionic gonadotrophin (β-HCG) on the results. Method: The study included 120 participants ranging in age from aged 18-45 years. Thirty of them had Missed abortion and Thirty had ectopic pregnancy were diagnosed by X-ray, whereas sixty were healthy and served as a control group. The Activin A and beta-HcG levels measured by enzyme linked immunosorbent assay method. Body mass index "BMI" was calculated by applying the continuity formula: - Body Mass Index (Kg/m2) = Weight/Height2. Results: Women with ectopic pregnancy and missed abortion had significantly higher levels of ACV-A and human chorionic gonadotropin beta (B-HcG) than the control groups. In addition, patients in the ectopic group had a significantly lower score than patients in the missed abortion group. Conclusion:Serum Activin A can be used as indicators of Ectopic pregnancy and Missed Abortion, Obesity represented by BMI has no significant effect on patients with ectopic pregnancy or missed abortion while Serum Beta-HcG level at cutoff value of >236, ng/ml) may be a novel biomarker for assessment of women with ectopic pregnancy .

Artesunate Alleviates Chronic Hyperoxia-induced Bronchopulmonary Dysplasia by Suppressing NF-κB Pathway in Neonatal Mice.

Bronchopulmonary dysplasia (BPD) is a chronic lung condition that occurs in premature infants who undergo prolonged mechanical ventilation and oxygen therapy. Existing treatment methods have shown limited efficacy, highlighting the urgent need for new therapeutic strategies. Artesunate (AS) is a compound known for its potential anti-inflammatory properties, and studies have shown its protective effects against acute lung injury. However, its impact on BPD and the underlying mechanisms remain unclear. To investigate the effect and underlying mechanism of AS on chronic hyperoxiainduced BPD in neonatal mice. Full-term C57BL/6J mice were randomly assigned to the Air+lactate Ringer's solution (L/R) group, O2 + L/R group, and O2 + AS group. Analysis was performed using assay methods such as ELISA, RT-qPCR, hematoxylin-eosin staining, and Western blotting. Compared with the O2+L/R group, the expression of inflammatory factors in the serum, tissue, and BALF of the O2+AS group was significantly reduced, the lung function of the mice was improved, and the inflammatory infiltrates were significantly alleviated. AS inhibited the mRNA expression of inflammatory factors in mice. We found that the expression of nuclear p65 and cytoplasmic p-IκBα in the NF-κB pathway was inhibited after adding AS. AS ameliorated chronic hyperoxia-induced BPD in neonatal mice probably by inhibiting the expression of NF-κB pathway and inflammatory factors.

Stability Indicating High‐performance Liquid Chromatography Method for Quantification of Amoxapine and Characterization of Forced Degradation Products Employing Quadrupole‐Time of Flight Mass Spectrometry

ABSTRACTAmoxapine, a tricyclic antidepressant, has been widely used for long‐term treatment of depression along with other psychiatric disorders involving neurosis, agitation, and anxiety. The drug may undergo degradation during manufacturing, transportation, and storage. To ensure drug safety, both identification and characterization of the degradation products (DPs) should be considered. Accordingly, a sensitive and selective stability‐indicating assay method was developed for amoxapine using high‐performance liquid chromatography (HPLC). The amoxapine drug was subjected to acidic hydrolysis, basic hydrolysis, and oxidation separately. The degradation susceptibility of the amoxapine was found to be in the order of Acid hydrolysis > Oxidation > Base hydrolysis. The developed method was validated as per the International Council for Harmonization Q2(R2) guideline. Further, LC‐electrospray ionization‐quadrupole‐time of flight‐tandem mass spectrometry was employed for the identification and characterization of the DPs formed under stress conditions. The study revealed that under accelerated stress conditions, three hydrolytic DPs and one oxidative DP were formed. Three of the four DPs were found to be novel and never reported before. In addition, a plausible mechanism representing the formation of all DPs has also been established.

The Rolled Towel Method for Hormone Response Assays in Maize.

The rolled towel assay (RTA) is a soil-free method to evaluate juvenile phenotypes in crops such as maize and soybean. Here, we provide an updated RTA-based protocol to phenotype maize seedling responses to chemicals of interest. We exemplify the protocol with two synthetic auxin herbicides (2,4-dichlorophenoxyacetic acid and picloram), an auxin precursor (indole-3-butyric acid), and an auxin inhibitor (N-1-naphthylphthalamic acid), but the method can be used with other hormones or plant growth regulators that are soluble in growth media. We also include instructions on how to annotate root traits and analyze primary root length trait data. The protocol can be scaled up for use in genetic screens, preparing tissue for gene expression analyses, carrying out genome-wide association studies (GWASs), and quantitative trait locus (QTL) identification.

Effects of cholecalciferol supplementation on depressive symptoms, C-peptide, serotonin, and neurotrophin-3 in type 2 diabetes mellitus: A double-blind, randomized, placebo-controlled trial

The coexistence of depression and type 2 diabetes mellitus (T2DM) can significantly worsen disease prognosis and lower quality of life. Emerging evidence suggests that vitamin D deficiency contributes to the progression of T2DM and is closely associated with the development of depression. The aim of this study was to investigate the effects of cholecalciferol on depression in patients with T2DM, exploring its mechanisms by analyzing its impact on C-peptide, serotonin, and neurotrophin-3 levels. A double-blind, randomized, placebo-controlled clinical trial was conducted at Cipto Mangunkusumo General Hospital, Jakarta, Indonesia, from April 2021 to September 2022. Patients with T2DM and depressive symptoms were randomly assigned to two groups: received 4000 IU of cholecalciferol daily and received a placebo for 12 weeks. Depression was assessed using the Beck Depression Inventory-II (BDI-II) before and 12 weeks after the intervention. The levels of C-peptide, serotonin, and neurotrophin-3 were measured at the end of the fourth week of intervention using the enzyme-linked immunosorbent assay (ELISA) method. Between-group comparisons were made using independent Student t-tests and Mann-Whitney U tests. Paired Student t-tests or Wilcoxon tests were applied for within-group comparisons between pre- and post-intervention. A total of 70 T2DM patients with depression were included in this study, comprising 38 patients in the cholecalciferol group and 32 in the placebo group. C-peptide levels increased significantly in the cholecalciferol group compared to the placebo group (p=0.006). No significant differences were observed in serotonin and NT-3 levels between the cholecalciferol group compared to the placebo group. The cholecalciferol group had a significantly greater reduction in BDI-II scores compared to the placebo group (p<0.001). This trial highlights that taking cholecalciferol might help ease mild to moderate depression symptoms in patients with T2DM by enhancing c-peptide levels, though its effects on serotonin and neurotrophin-3 are still unclear.

The Effects of Traditional Asian Diet on Metabolism, Gut Microbiota, and Liver Tissue in NASH Rats

Traditional Asian Diets (AD) in rural areas have a significant risk of mortality due to cirrhosis and hepatocellular carcinoma as nonalcoholic fatty liver disease (NAFLD). This study aims to determine the relationship between AD and liver cancer cases using rat experimental animals Rattus norvegicus strain Wistar. The measured variables include metabolic parameters, gut microbiota profile, and liver histology. This study used 14 rats in two groups: Chow Diet (7 rats to CD) and AD (7 rats to AD), and were given the respective diets for 12 weeks. Enzyme-linked immunosorbent assay (ELISA) methods are used to analyze liver enzymes, lipid profiles, and blood sugar levels. The analysis of gut microbiota used variable region-specific 16S rRNA gene and V3-V4. Biopsy stained with Hematoxylin Eosin was used to study the histology of the liver. Moreover, it was analyzed utilizing NAS (NAFLD Activity Score). The result of this study indicated that reduce body weight the rats treated with AD significant different than treated with CD. Firmicutes, Lactobacillus reuteri, Prevotellaceae bacterium, Romboutsia ilealis, and Bacteroidota in AD greater than CD. Alzheimer's disease had notably higher levels of alkaline phosphatase compared to those diagnosed with Crohn's disease on individual diagnosis. Differences in total bilirubin, alanine transaminase, aspartate transaminase, blood sugar, total cholesterol, high-density lipoprotein, low-density lipoprotein, and triglycerides were not significant. The NAS analysis indicated that the two groups comprised rats lacking non-alcoholic fatty liver disease. Despite the high caloric content of the Asian diet, it did not lead to significant changes in metabolic parameters and liver histology related to non-alcoholic steatohepatitis. This behavior can be ascribed to the advantageous influence of the gut microbiota.

Unveil the molecular recognition mechanisms of triazine haptens and monoclonal antibodies: Establishing ic‐ELISA methods for triazine herbicide analysis in tea

AbstractThis study introduced two triazine haptens (with 0 or 1 carbon connecting arms at nonisopropyl positions) and developed two heterologous indirect competitive enzyme‐linked immunosorbent assay (ic‐ELISA) methods. The IC50 values for the 17 triazine pesticides ranged from 0.84 ng/mL to 225.63 ng/mL, and the established method was applied to determine their concentrations in tea samples. This study aimed to address the current debate on the recognition mechanism of triazine haptens with shorter connecting arms. This study produced two new findings by investigating the recognition mechanism of triazine haptens and antibodies. On the one hand, the specific amino acid isoleucine (ILE‐219) is crucial for isopropyl recognition. On the other hand, the selection of the connecting site of the triazine hapten has a significant effect on the specificity of the antibody, with the influence of the length of the carbon connecting arm being secondary. Our newly designed haptens show promise for diverse triazine pesticide immunoassay applications. This discovery established a foundational framework for constructing a tailored antibody library for pollutants.

Ameliorative effects of osthole on acrylamide-induced neurotoxicity in PC12 cells: Role of oxidative stress, apoptosis and ERK pathways.

The possible protective effects of osthole on acrylamide-induced neurotoxicity in PC12 cells. Cells were pretreated with different concentrations of osthole (1- 25μM) for 24h and then the IC50 value of acrylamide (5mM) was added. After 24h, cell viability and intracellular ROS content were detected by MTT assay and DCF-DA methods, respectively. Also, DNA fragmentation in apoptotic cells was determined by propidium iodide assay, and apoptosis (Caspase-3, Bax, Bcl-2, ERK, and P-ERK) was measured by the western blot method. Exposing PC12 cells to acrylamide diminished cell viability, and enhanced the intracellular ROS generation and the percentage of apoptotic cells. Furthermore, acrylamide elevated the P-ERK/ERK and Bax/Bcl-2 ratio, and the level of cleaved caspase-3 protein in PC12 cells. Pretreating cells with osthole enhanced cell viability and reduced ROS generation. Also, osthole (10μM) significantly reduced P-ERK/ERK and Bax/Bcl-2 ratio, the level of cleaved caspase-3 protein, and the percentage of apoptotic cells in comparison to the acrylamide group. Osthole can exhibit a protective effect on the neurotoxicity of acrylamide through the inhibition of oxidative stress and apoptosis in PC12 cells.

The Role of Nanopartikel Green Tea in Enhancing Endothelial Cell Migration in HUVEC Culture exposed to EPC-Conditioned Media in Hyperglycemic Conditions

Backgrounds: The role of endothelial progenitor cells (EPCs) in angiogenesis is impaired in diabetes mellitus. The present study was conducted to observe the effects of nanopartikel green tea on the ability of EPCs exposed to high levels of glucose to release NO and induce endothelial cell migration. Purpose: This study aims to describe the mechanism of the role of green tea nanoparticles in increasing migration of endothelial cells in HUVEC cultures exposed to EPC-CM. The function of EPCs was assessed by evaluating HUVEC migration after administration of EPC-Conditioned Media. HUVEC migration was assessed by the Boyden chamber assay method. Method: Green Tea Nano Particles treatment at doses of 30 nM and 50 nM significantly increased endothelial cell migration at high glucose exposure of 22 mM. At a dose of 50 nM, normal glucose exposure increased endothelial cell migration better than high glucose exposure. Administration of high levels of glucose resulted in a decrease in the ability of EPCs to induce HUVEC migration, decrease in NO of EPCs, and decrease in the factor affecting EPC migration, namely SDF-1α and CXCR4. Results: Administration of 50 μM of nanopartikel green tea could inhibit the decreased ability of EPCs to induce HUVEC migration, and this effect was associated with NO, SDF-1α and CXCR4 concentrations. Conclusions: Administration of nanopartikel green tea could maintain the ability of EPCs exposed to high levels of glucose to release NO and induce HUVEC migration through an increase in SDF-1α and CXCR4.

The expression of sICAM-1 influenced by Clonorchis sinensis co-infection in CHB patients.

Soluble Intercellular Adhesion Molecule-1 (sICAM-1) has emerged as an inflammatory biomarker of many essential functions. We investigated the level of sICAM-1 influenced by Clonorchis sinensis (C. sinensis) co-infection in chronic hepatitis B (CHB) patients to explore the degree of liver tissue inflammation and liver function damage after co-infection. The study included data from patients with C. sinensis mono-infection (n=27), hepatitis B virus (HBV) mono-infection (n=32), C. sinensis and HBV co-infection (n=24), post-hepatitis B liver cirrhosis (n=18), post-hepatitis B liver cirrhosis co-infected with C. sinensis (n=16), and healthy controls (n=39). The level of sICAM-1 was measured with specific enzyme-linked immunosorbent assay method. Compared to the healthy control group, all the experimental groups had significantly higher serum sICAM-1 levels. The levels of sICAM-1 in co-infected groups were significantly higher compared to the mono-infection groups and were positively correlated with the levels of glutamate aminotransferase (ALT) and aspartate aminotransferase (AST). Our research findings confirmed that co-infection could exacerbate liver tissue inflammation and liver function damage in patients, could raise the sICAM-1 level, and may lead to the chronicity of HBV infection. These results provide clues for pathological mechanism study and formulating treatment plans.

Alamar Blue assay optimization to minimize drug interference and inter assay viability

Alamar Blue (AB) has become an increasingly popular reagent of choice for cell viability assays. We chose AB over other reagents such as MTT and Cell-Titer Glo due to its cost effectiveness and its ability to be a nondestructive assay. While analyzing the effect of osimertinib, an EGFR inhibitor on the non-small cell lung cancer cell line, PC-9, we noticed unexpected right-shifts of dose response curves as compared to the curves obtained by Cell Titer Glo assay. Here, we describe our modified AB assay method to avoid that right shift in dose response curves.•Removal of the drug containing medium prior to AB addition eliminated the falsely increased readings, giving comparable dose response curves as determined by Cell Titer Glo assay.•Plate-to-plate variability can be minimized by adding an appropriate concentration of a common fluorescence calibration standard to the assay plates to calibrate fluorimeter sensitivity.•Alamar Blue can be used as a continuous longitudinal assay to monitor cell growth or recovery from drug toxicity over time.

Lipoprotein(a) levels in acute myocardial infarction patients and their associations with prior events and coronary artery disease severity: a real-world cohort study

Abstract Introduction Circulating lipoprotein(a) [Lp(a)] has been associated with the risk of atherosclerotic coronary artery disease (CAD) and is an emergent therapeutic target. The objective of this study is to explore the association between serum levels of Lp(a) and previous myocardial infarction (MI) and/or coronary revascularization and the extension of obstructive CAD in patients with acute MI undergoing coronary angiography. Methods All consecutive patients with MI who underwent coronary angiography and Lp(a) measurement between May and November 2023 were included. Patient data were either registered prospectively or completed retrospectively using electronic health records. Lp(a) was measured using the Immunoturbidimetric Assay method with reaction intensification by particles, employing the World Health Organization/International Federation of Clinical Chemistry and Laboratory Medicine (WHO/IFCC) International Reference Reagent (SRM2B), on the "Roche Cobas C702" chemistry analyzer. The number of major coronary arteries (left main, left anterior descending, left circumflex and right coronary arteries) with stenosis >50% on the coronary angiography were registered. In the primary analysis we explored the association between Lp(a) levels using tertiles and previous occurrence of MI/revascularization, as well as the extension of obstructive CAD. In the secondary analysis we compared Lp(a) levels in patients with and without previous MI/revascularization and patients with 3-4 major vessels CAD vs. 0-2 vessels. Results A total of 116 patients were enrolled, with a mean age of 62±13 years, 79% males, 29% with known coronary artery disease, 71% with dyslipidemia, 24% with diabetes, 65% with hypertension, and 58% were either active or former smokers. The median Lp(a) level was 72 (20-183) nmol/L. In the primary analysis (table 1), the number of previous MI/revascularizations was significantly higher in the third tertile of Lp(a) (T3) versus T1 (p=0.044). No significant differences were found for the remaining analyses. In the secondary analysis (figure 1) the level of Lp(a) was significantly higher in patients with 3-4 vessel CAD vs. 0-2 vessel (158.4±145.2 nmol/L vs. 103.16±104.6 p=0.044). Conclusion In this small real-world cohort population with acute MI, the number of previous MI or coronary revascularization was higher in patients with increased levels of Lp(a) and this marker of atherosclerotic disease was greater in patients with more extensive CAD.

Biofilm formation and drug susceptibility of biofilm Candida spp. clinically isolated from nasopharyngeal cancer patients in Vietnam

Background and Objectives: The biofilm formation has been widely recognized as one of the main mechanisms of antimi- crobial resistance development in microorganisms. However, few studies are focusing on this phenomenon in Candida spp. in clinical settings, especially on immuno-compromised patients. Materials and Methods: In this study, both the rate of biofilm formation in those patients and its drug susceptibility in initial and mature biofilm were assessed using crystal violet assay and dilution method. Results: The results demonstrated that the biofilm formation rate was similar between albicans and non-albicans Candida. However, the biofilm formation capacity was more pronounced in non-albicans Candida, especially, C. glabrata. As expect- ed, there was a significant relationship between biofilm formation and drug resistance. In addition, our study reconfirmed that the age of high concentration of antifungal agents only affected Candida before its biofilm formation regardless of its biofilm formation capacity. In the contrary, once the biofilm was formed even elevated drug concentrations did not show sufficient efficacy, highlighting a need for high dosage at the early stage of treatment for those patients. Conclusion: The results of this study highlighted the importance of using appropriate antifungal agents for Candida treatment before the formation of biofilm.

Phytochemical composition, in vitro cytotoxicity, and in silico docking properties of Tamarix tetragyna L.

Tamarix tetragyna is a plant grows in Mediterranean area and some Arab countries. It possesses numerous medicinal values. Purpose of our study is to explore biological activity of tamarix tetragyna extracts of both leaves and stem with investigating their phytochemical composition. The investigated extracts’ phyto-constituent composition was determined using gas chromatographic-mass spectrometric method. In addition, in vitro cytoxicity activity versus cancer cell lines such MCF-7, HepG-2, HCT-116, and A-549 was examined by MTT assay method, together with exploring its apoptosis effect by flow cytometry and western blot analysis techniques. Moreover, some phytochemical compounds were identified, and in-silico evaluated against anticancer molecular targets. Plant extracts showed good cytotoxic activity against both A-549 and HCT-116 cancer cell lines. With an IC50 value of 23.90 µg/ml that led to apoptosis and G2/M-phase arrest in A-549 cells, cytotoxicity data demonstrate leaves’ extract effectiveness against these cells. Upon GC-MS analysis, it revealed presence of some bioactive components such as Stigmast-5-en-3-ol and 2-methoxy-4-vinyl phenol, which are known for their cytotoxic activity. Our findings suggest that methanolic extracts of Tamarix tetragyna parts may have potential therapeutic uses as anticancer against A-549 cells, which opens up further avenues for investigation into its industrial applications.

Novel Strategies for Synthesis and Assessment of Dextran Conjugated 9Z,11E Linoleic Acid on Cancer Cell Lines Viability via Wnt/Beta‐Catenin Signaling Pathway and Apoptosis

AbstractConjugated linoleic acid (CLA) is one of the positional and structural isomers of linoleic acid. Ninety percent of the CLA contained in natural foods is the cis‐9(9Z), trans‐11(11E) isomer. CLA also has anti‐carcinogenic, anti‐adipogenic, and anti‐inflammatory properties. Limited studies aim to elucidate the impact of 9Z,11E‐CLA on Wnt/beta‐catenin signaling.In this study, we have selected MDA‐MB‐231, T98G, and SAOS‐2 cell lines of various cancer origins and investigate the impact of dextran conjugated 9Z,11E CLA (Dex‐9Z,11E‐CLA) using cell proliferation assay and immunocytochemical methods. Cytotoxic effects of Dex conjugated 9Z,11E CLA are assessed with the MTT method. It analyzed Wnt/beta‐catenin signaling changes using target molecule beta‐catenin, Dickkopf‐1 (Dkk‐1) as a negative regulator, and active caspase3 (CC3) as an apoptotic marker.Dex‐9Z,11E‐CLA is shown to suppress cell viability in three different types of cancer cells, and the most effective concentration is 200 µm. 9Z,11E‐CLA and Dex‐9Z,11E‐CLA affect the Wnt/beta‐catenin signaling pathway negatively in all cell lines. Beta‐catenin expression is decreased, Dkk‐1 expression is increased, and CC3 positivity is increased significantly compared to the control group. In conclusion, the data suggest that Dex‐9Z,11E‐CLA suppresses cell viability and causes apoptosis in cells via the Wnt/beta‐catenin signaling pathway. Lastly, Dex‐9Z,11E‐CLA harbors novel anticancer potential and exhibits uncontrolled activation of Wnt/beta‐catenin signaling.

Self-assembly of S,N-codoped Ce/Cu bimetallic nanoparticles for fluorescence and visual detection of hexavalent chromium.

Ce2(SO4)3 was doped into 4,6-diamino-2-mercaptopyrimidine (DAMP)-encapsulated copper nanoclusters (CuNCs) via a facile, rapid, low-temperature, and green self-assembly synthesis method to obtain fluorescent S,N-codoped Cu/Ce-DAMP nanoparticles (S,N-codoped Cu/CeNPs) for the detection of Cr(VI). The prepared Cu/CeNPs exhibit double emission peaks at 470nm and 610nm. Thefluorescence emission at 610nm is significantly enhanced due to the aggregation-induced emission (AIE) effect, and the quantum yield is as high as 20.19%. The fluorescence emission at 610nm can be selectively quenched by Cr(VI) due to the internal filter effect (IFE) and dynamic quenching, whereas the weak fluorescence at 470nm remains almost stable. On this basis, a fluorescence assay method for Cr(VI) was established, with good linearity in the concentration range 0.5-120µM and a detection limit (LOD) of 134nM. Using a smartphone to take photos of the fluorescence signals changes caused by Cr(VI) rapid visual detectionis achieved with a linear range of 10-130 μM and a LOD of 2.35 μM. The proposed method was successfully applied to the detection of Cr(VI) in actual water samples.

In Vitro Antioxidant and Antifungal Activities of Extracts from Ocimum basilicum Leaves Validated by Molecular Docking and ADMET Analysis.

The aim of study is to investigate the various mechanism-based antioxidant and anti-fungal properties of a hydroalcoholic extract of Ocimum basilicum L leaves. Additionally, conduct molecular docking to simultaneously validate in vitro activities. Also, perform ADMET analysis to know pharmacokinetic properties and its toxicity for its safety. Prior extract's qualitative analysis has been performed to identify the bioactive compounds by phytochemical tests and GC-MS analysis. Different mechanism-based in vitro antioxidant methods are studied; in different methods, different IC50 values have come, which revealed the extract's antioxidant potentials. The antifungal potential of the extract has been observed by performing a modified poison food assay and a time-killing curve assay. In silico analysis with the human peroxiredoxin 5 enzyme (PDB ID: 1HD2) and secreted aspartic proteinase (PDB ID: 2QZX), which predict extract biological activity, has shown promising results of Ocimum basilicum L extract. In silico findings confirm the in vitro experimental outcome of the extract. The different IC50 values of extracts in different mechanisms indicate their therapeutic potential, and that encourages further research in formulation development. The time-killing assay method gives information about a dynamic interaction between extract and microbial strain. Concentration-dependent antifungal studies have significance in formulation development for dose determination.