Assay Technique Research Articles

Circadian disruption and psychostimulants dysregulates plasma acute-phase proteins and circulating cell-free mitochondrial DNA

BackgroundPrevious studies have indicated a close link between the inflammatory response, exacerbated by circadian disruption and psychostimulants such as cocaine and methamphetamine (METH). Indicators of this inflammation include cortisol and acute-phase proteins (APPs) like C-reactive protein (CRP), complement C3 (C3), and serum amyloid A (SAA). The connection between these inflammation markers and circulating mitochondrial DNA (mtDNA) has been gaining attention. However, the specific influence of cocaine and METH on APP, cortisol, and mtDNA levels in mice with disturbed circadian rhythm has yet to be explored, which is the main aim of this research. MethodsIn our study, we employed 10–12-week-old male C57BL/6J mice, which underwent an imposed 6-h phase advance every six days for a total of eight cycles. This process led to the formation of mice with disrupted circadian rhythm and sleep disorders (CRSD). We administered 11 dosages of cocaine and METH 15 mg/kg and 20 mg/kg, respectively to these CRSD mice over the course of 22 days. Quantitative assessments of CRP, C3, SAA, cortisol, and cell-free circulating mtDNA were conducted using enzyme-linked immunosorbent assay (ELISA), Western Blot, and quantitative real-time polymerase chain reaction (qRT-PCR) techniques. ResultsThe experiment revealed that disruption in circadian rhythm alone or cocaine or METH on their own increased CRP, C3, SAA, and cortisol levels in comparison with the control group. CRSD mice, exposed to cocaine and METH, showed a significant rise in CRP, C3, and SAA, while those without exposure remained stable. We also found a reduction in circulating cell-free mtDNA in all CRSD mice, regardless of cocaine and METH exposure. ConclusionsThe findings of our study affirm that the levels of CRP, C3, SAA, and cortisol, which reflect inflammation, are enhanced by circadian disruption, cocaine, and METH, and these levels show a strong correlation with the content of circulating cell-free mtDNA. Furthermore, it also shows the potential link between the disruption of the circadian clock and the inflammatory response triggered by cocaine and METH.

Concentration of Asprosin Associated with Poor Control of Type 2 Diabetes Mellitus in Diyala Province

A newly secreted adipokine called asprosin is brought on by fasting as well as encourages hepatic glucose release. Its loss of function through immunologic or genetic methods has a significant effect on lowering glucose and insulin as a result of decreased hepatic glucose release. This study aimed to measure the level of asprosin in a group of poor control hyperglycemic patients and compare its level with the control group and find its correlation with obesity and lipid profile. Asprosin level in serum was assessed by enzyme-linked immunosorbent assay (ELISA) technique for 110 Iraqi patients with type 2 diabetes mellitus(T2DM), as well as 70 individual healthy controls. Percentage of glycated hemoglobin (HbAIc), fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL) and low-density lipoprotein (LDL) concentration were calculated using Roche cobas integra 400 plus. Patients with type 2 diabetes mellitus reported increased levels of asprosin in their blood when compared with controls (p < 0.001). A significant positive correlation was seen between asprosin and FBG in patient’s (P< 0.05). Non-significant correlation was seen between asprosin and body mass index (BMI) or lipid profile. In conclusion, according to the asprosin level in serum, there was a highly significant difference between patients with type 2 diabetes mellitus and control group. BMI and lipid abnormalities were not correlated to asprosin level in the serum of patients.

Urotensin II promotes the proliferation and secretion of vascular endothelial growth factor in rat dermal papilla cells by activating the Wnt-β-catenin signaling pathway

Introduction. Urotensin II (U II) is a kind of active peptide with a variety of biological effects, such as promoting cell proliferation and endocrine effects. The aim of this study is to investigate the effect of urotensin II on the proliferation and secretion of vascular endothelial growth factor (VEGF) in cultured rat dermal papilla cells (DPCs), and to explore its molecular mechanism. Materials and Methods. We used the DPCs isolated from the thoracic aortas of Wistar-Kyoto rats to run the CCK8 and ELISA assay, RC-PCR and Western blotting techniques to identify the effect of Urotensin II on the proliferation and secretion of VEGF in DPCs, data were analyzed by one-way ANOVA or t-test. Results. U II can increase the mRNA expression of proliferation markers Ki67 and PCNA. In addition, the Wnt/β-catenin pathway was activated by U II, but Wnt inhibitor DKK1 reversed the effect of U II. Conclusions. U II promoted the proliferation and secretion of VEGF in rat DPCs through activation of the Wnt-β-catenin signaling pathway.

A multi-unit integrated electrochemical biosensor array for synergistic signal enhancing carbohydrate antigen 125 detection

Significant progress has been made in recent decades; however, the development of an accurate, portability and high-throughput electrochemical biosensor for quantifying trace biomarker in complex biological samples remains challenging. In this study, we described a novel 3 × 3 multi-unit portable integrated biosensor array, specifically designed for parallel and precise detection of trace levels of carbohydrate antigen 125 (CA-125) using multiplex synergistic signal amplification. The sensor array substrate was constructed on a planar printed circuit board (PCB) using printing technology, making them integrated, portable, affordable and disposable. The exceptional sensitivity of the biosensor is attributed to the nitrogen-doped mesoporous carbon (NMC) nanomaterial, which possesses a large surface area, superior conductivity and tunable pore size, enabling high affinity of and enzyme catalytic activity. The integrated biosensing array platform consists of individually controlled sensors, allowing for high-throughput tests of the same sample simultaneously, ensuring the reliability and accuracy of assay. The developed biosensor array system demonstrated excellent sensitivity and rapid CA-125 assay, achieving an impressive detection limit as low as 0.027 U/mL (S/N = 3). Meanwhile, the high-throughput multiplex test results (n = 9) were obtained within a short timeframe of 30 min. Ultimately, the biosensor array successfully quantified CA-125 in human serum, yielding results comparable to the enzyme–linked immunosorbent assay technique. This technology presents a highly promising strategy that can be potentially extended to detect other biomarkers for biological analysis and early disease diagnosis.

ELISA F29 -A therapeutic efficacy biomarker in Chagas disease: Evaluation in pediatric patients treated with nifurtimox and followed for 4 years post-treatment.

Measurement of the success of antitrypanosomal treatment for Chagas disease is difficult, particularly in the chronic phase of the disease, because anti-Trypanosoma cruzi antibodies persist in serum for prolonged periods. We studied the effects of nifurtimox administered by two different treatment regimens on the T. cruzi calcium-binding flagellar protein F29 in children diagnosed with Chagas disease measured using an enzyme-linked immunosorbent assay (ELISA) technique (ELISA F29). In a phase 3, randomized, double-blind, parallel-group, historically controlled study (ClinicalTrials.gov NCT02625974), blood samples obtained from children diagnosed with Chagas disease and treated with nifurtimox for either 60 days or 30 days were analyzed using an ELISA with an F29 recombinant protein as the antigen, as well as conventional serological tests (recombinant ELISA and indirect hemagglutination assay). In an exploratory approach, serological response to nifurtimox treatment was evaluated for 4 years post-treatment. In both treatment groups, the number of patients with negative ELISA F29 values increased over the period of observation. The incidence rate of negative seroconversion using ELISA F29 was 22.94% (95% CI: 19.65%, 26.63%) in the 60-day treatment group and 21.64% (95% CI: 17.21%, 26.86%) in the 30-day treatment group. In the subpopulation of patients who tested seropositive for F29 before nifurtimox treatment, 88 patients (67.7%) in the 60-day regimen and 39 patients (59.1%) in the 30-day regimen were F29 seronegative at 4 years post-treatment. All patients who had a positive ELISA F29 test at baseline and seroconverted to negative measured by conventional serology reached seronegativity in ELISA F29 earlier than in conventional serology. The results demonstrate a serological response to treatment with nifurtimox measured by the ELISA F29 test in children diagnosed with Chagas disease. The F29-based ELISA can be considered a potential early marker of response to antitrypanosomal therapy for Chagas disease. ClinicalTrials.gov NCT02625974.

1694-P: Impact of Bariatric Surgery on Increased Circulating Levels of Several Biomarkers for Neurodegenerative Diseases in Obese Chinese

Background: Increasing studies have shown a strong association of obesity with neurodegenerative disorders. However, there is scarce evidence for the direct link between obesity and neurodegeneration. In addition, although bariatric surgery is well-known to be an effective therapy for obesity-related metabolic dysfunctions, its impact on cognitive function remains uncertain. This study aims to address these questions by monitoring several well-established circulating biomarkers for neurodegeneration in morbidly obese subjects receiving bariatric surgery. Methods: The single molecule array, an ultra-sensitive enzyme-linked immunosorbent assay technique, was used for measurement of plasma levels of p-tau 181, tau, Aβ42 and Aβ40 in 36 morbidly obese individuals (BMI=39.46±7.87) before and after bariatric surgery for 12 months and 55 lean subjects (BMI=21.79±2.84). Results: Plasma levels of p-tau 181, tau, Aβ42 and Aβ40 in obese individuals, were significantly elevated compared to lean individuals, and positively correlated with several indices of obesity, including body weight, BMI and waist circumference. In obese subjects who received bariatric surgery, significant improvements on many metabolic parameters were observed, such as BMI, HOMA-IR and triglyceride. However, circulating p-tau 181 levels in obese individuals kept unchanged before and after treatment, while tau, Aβ42 and Aβ40 levels increased from 4.34±0.74 to 6.06±0.38 pg/ml, 4.37±0.51 to 8.47±0.41 pg/ml, and 98.89±11.53 to 174.14±8.99 pg/ml, respectively. Conclusions: Circulating levels of several neurodegenerative biomarkers are elevated in obesity but such changes cannot be reversed by bariatric surgery. These findings implicate the adverse effects of obesity at the early stage of life on neurodegeneration may be last-lasting and hardly repairable, highlighting the importance of maintaining a healthy weight in prevention of neurodegenerative diseases. Disclosure R.Yang: None. H.Li: None. W.Yang: None. Q.Wu: None. L.Jin: None. E.Song: None. C.Wang: None. A.Xu: None. Funding Hong Kong Research Grants Council (AoE/M/707-18)

Screening for antimicrobial residues in poultry eggs in Bangladesh using Charm II radio-receptor assay technique following validation

ABSTRACT The aim of the study was to screen for the presence of antimicrobial residues in poultry eggs from Bangladesh using the Charm II radio-receptor assay in the absence of expensive confirmatory instrumentation. This was based on cut-off values as set in the validation guidelines according to Commission Decision 2002/657/EC and Commission Implementing Regulation (EU) 2021/808. Fortified eggs spiked with fixed concentrations of doxycycline, erythromycin A, sulphamethazine, and benzylpenicillin were used to determine the cut-off values and detection capabilities (CCβ). Other validation parameters included were applicability, ruggedness, and robustness. A total of 201 egg mix samples from native organic chicken, duck, and commercial farm-raised laying hens (both brown and white eggs) were tested and after analysis 13%, 10%, and 4.5% of the egg mix samples showed positive signals for sulphonamides, macrolides/lincosamides, and tetracyclines, respectively. Presence of multiple drug residues were also suspected in 11 out of 201 egg mix samples.

Neutron resonance fission neutron analysis for nondestructive fissile material assay

We propose neutron resonance fission neutron analysis (NRFNA), an active nondestructive assay (NDA) technique, to improve the capability to identify and quantify a small amount of fissile material in a sample. NRFNA uses pulsed neutrons to induce fission reactions in the sample. Fission neutrons are detected by a neutron-gamma pulse shape discrimination (PSD) scintillation detector with time-of-flight (TOF) technique. The obtained nuclide-specific resonance peaks in the neutron energy spectrum provide information to identify and quantify a fissile material in the sample. The possibility of using PSD for NRFNA was confirmed through a test experiment using a natural uranium sample. We successfully observed the resonance peaks from 235U(n,f) reaction and showed that NRFNA would be useful for measuring a small amount of fissile material in a sample.

Metformin Effects on Blood Levels of Myonectin in Polycystic Ovarian Women

Background: Polycystic ovary syndrome (PCOS) is a most prevalent endocrine diseases for females of childbearing age, and lead to an ovulatory infertility. It was shown that myonectin may be positively associated with insulin resistance parameters. Objectives: To evaluate serum myonectin levels and to determine the effects of metformin treatment on myonectin levels in patients with polycystic ovary syndrome. Patients and Methods: The Cross-Sectional Study carried out in the Department of Obstetrics and Gynecology Salahdeen general hospital in Tikrit city from 1st November 2022-30th January 2023. Patients diagnosed with PCOs depending based on the Rotterdam criteria. Participants were enrolled after all eligibility criteria were confirmed and informed consent completed. Sixty PCOS patients were selected as the PCOS group, while 30 healthy women matched for age with the PCOS patients were selected as the control group. Only 30 patients of them complete the follow up study and they agree to continue on metformin treatment during three months, the duration of the follow up. They take metformin 850 mg twice daily for three months and provide fasting blood samples on the second day of menstruation before and after treatment. The data collection done through: a designed closed and open-ended questionnaire, by using direct interviewing and Ultrasound examination, Laboratory examination. Myonectin, Luteinizing hormone (LH), Follicular Stimulating Hormone (FSH), and insulin were analysed by enzyme-linked immunosorbent assay (ELIZA) technique from Biomeriuex. Data were analyzed using SPSS for Windows 7. Results: The level of myonectin decreased significantly in PCOS patients compare to control group. The level of blood glucose, Insulin, and HOMA-IR, were increased significantly compared to the control. Serum LH levels were significantly higher, while the level of FSH were lowerin in women with PCOS than in controls. After metformin intake, all patients showed significant in decrease in gremlin concentration at p-values 0.05, but no significant difference (P>0.05) in myonectin level when compared to pre-treatment. Treatment resulted a significant decrease in in body mass index, Blood glucose, Insulin, HOMA-IR, testosterone and LH at p-values 0.05. However, the study found no significant difference (p > 0.05) in myonectin level between both groups in treated group with metformin compared with pre-treatment. Conclusion: Metformin testosterone and insulin resistance but can’t induce changes of myonectin level in patients with polycystic ovary syndrome.

Inflammatory biomarkers differentiate the stage of maturation in chronic subdural hematomas

ObjectiveInflammation is a major pathophysiological driver of the development of chronic subdural hematomas (CSDH), but there is still limited knowledge on the key molecular processes and corresponding biomarkers involved in this disease. In this study, the aim was to study a subset of inflammatory biomarkers and their relation to the clinical status of the patient and the radiological characteristics of the CSDH. MethodsIn this observational study, 58 patients who were operated on with CSDH evacuation, at the Department of Neurosurgery, Uppsala, Sweden, between 2019 and 2021, were prospectively included. The CSDH fluid was collected peri-operatively and was later analyzed with proximity extension assay (PEA) technique (Olink) for a panel of 92 inflammatory biomarkers. Demographic, neurological (Markwalder), radiological (general (Nakaguchi classification) and focal (septa below the burr holes)), and outcome variables were collected. ResultsIn 84 of the 92 inflammatory biomarkers, the concentration was above the detection limit in >50% of the patients. There was a significant difference in GDNF, NT-3, and IL-8 depending on the Nakaguchi class, with higher values in the trabeculated CSDH subtype. In addition, those with septa at the focal area of CSDH collection, had higher levels of GDNF, MCP-3, NT-3, CXCL1, CXCL5, IL8, and OSM. There was no association between Markwalder grade and the inflammatory biomarkers. ConclusionsOur findings support the presence of local inflammation in the CSDH, a shift in biomarker pattern as the CSDH matures towards the trabeculated state, potentially differences in biomarker patterns within the CSDH depending on the focal environment with presence of septa, and that the brain might develop protective mechanisms (GDNF and NT-3) in case of mature and long-standing CSDHs.

AATF inhibition exerts antiangiogenic effects against human hepatocellular carcinoma.

Angiogenesis is a key factor in the growth and metastasis of hepatic tumors and thus a potential therapeutic target in hepatocellular carcinoma (HCC). In this study, we aim to identify the key role of apoptosis antagonizing transcription factor (AATF) in tumor angiogenesis and its underlying mechanisms in HCC. HCC tissues were analyzed for AATF expression by qRT-PCR and immunohistochemistry. Stable clones of control and AATF knockdown (KD) were established in human HCC cells. The effect of AATF inhibition on the angiogenic processes was determined by proliferation, invasion, migration, chick chorioallantoic membrane (CAM) assay, zymography, and immunoblotting techniques. We identified high levels of AATF in human HCC tissues compared to adjacent normal liver tissues, and the expression was found to be correlated with the stages and tumor grades of HCC. Inhibiting AATF in QGY-7703 cells resulted in higher levels of pigment epithelium-derived factor (PEDF) than controls due to decreased matric metalloproteinase activity. Conditioned media from AATF KD cells inhibited the proliferation, migration, and invasion of human umbilical vein endothelial cells as well as the vascularization of the chick chorioallantoic membrane. Furthermore, the VEGF-mediated downstream signaling pathway responsible for endothelial cell survival and vascular permeability, cell proliferation, and migration favoring angiogenesis was suppressed by AATF inhibition. Notably, PEDF inhibition effectively reversed the anti-angiogenic effect of AATF KD. Our study reports the first evidence that the therapeutic strategy based on the inhibition of AATF to disrupt tumor angiogenesis may serve as a promising approach for HCC treatment.

Akkermansia muciniphila and its membrane protein ameliorates intestinal inflammatory stress and promotes epithelial wound healing via CREBH and miR-143/145

BackgroundThe intestinal epithelial barrier is the interface for interaction between gut microbiota and host metabolic systems. Akkermansia muciniphila (A. muciniphila) is a key player in the colonic microbiota that resides in the mucus layer, whose abundance is selectively decreased in the faecal microbiota of inflammatory bowel disease (IBD) patients. This study aims to investigate the regulatory mechanism among A. muciniphila, a transcription factor cAMP-responsive element-binding protein H (CREBH), and microRNA-143/145 (miR-143/145) in intestinal inflammatory stress, gut barrier integrity and epithelial regeneration.MethodsA novel mouse model with increased colonization of A muciniphila in the intestine of CREBH knockout mice, an epithelial wound healing assay and several molecular biological techniques were applied in this study. Results were analysed using a homoscedastic 2-tailed t-test.ResultsIncreased colonization of A. muciniphila in mouse gut enhanced expression of intestinal CREBH, which was associated with the mitigation of intestinal endoplasmic reticulum (ER) stress, gut barrier leakage and blood endotoxemia induced by dextran sulfate sodium (DSS). Genetic depletion of CREBH (CREBH-KO) significantly inhibited the expression of tight junction proteins that are associated with gut barrier integrity, including Claudin5 and Claudin8, but upregulated Claudin2, a tight junction protein that enhances gut permeability, resulting in intestinal hyperpermeability and inflammation. Upregulation of CREBH by A. muciniphila further coupled with miR-143/145 promoted intestinal epithelial cell (IEC) regeneration and wound repair via insulin-like growth factor (IGF) and IGFBP5 signalling. Moreover, the gene expressing an outer membrane protein of A. muciniphila, Amuc_1100, was cloned into a mammalian cell-expression vector and successfully expressed in porcine and human IECs. Expression of Amuc_1100 in IECs could recapitulate the health beneficial effect of A. muciniphila on the gut by activating CREBH, inhibiting ER stress and enhancing the expression of genes involved in gut barrier integrity and IEC’s regeneration.ConclusionsThis study uncovers a novel mechanism that links A. muciniphila and its membrane protein with host CREBH, IGF signalling and miRNAs in mitigating intestinal inflammatory stress–gut barrier permeability and promoting intestinal wound healing. This novel finding may lend support to the development of therapeutic approaches for IBD by manipulating the interaction between host genes, gut bacteria and its bioactive components.

Isolation and characterization of three bacteriophages infecting Erwinia amylovora and their potential as biological control agent

BackgroundFire Blight, incited by Erwinia amylovora, is one of the most damaging pear and apple diseases in the world. Fire blight was introduced to Egypt in the 1960 and threatens the Egypt’s costs for pear industry. Currently, Phage therapy is considered to be secured biological method for controlling plant bacterial diseases. This investigation aimed to isolate and identify molecularly for bacteria causing fire bright disease. As well as isolation and identification bacteriophages via spot and plaque assay techniques from pear fire blight lesions and soil. On the other hand, bacteriophages were identified based on plaque morphology, virion morphology, physical characters, profile of DNA restriction and protein.ResultsPathogenicity test revealed that healthy seedlings and pear fruits were responsive to fire blight E. amylovora. Considering the relatively wide host range and greatest protein and genetic variability, using restriction enzyme pattern, the three diversity phage isolates named, EAP1, EAP2 and EAP3 showed a lack of diversity out of five were fatherly characterized. The phages confirmed the close relation of EAP1, EAP2 to Siphoviridae (hexagonal head and long flexible non-contractile tail) and EAP3 to Myoviridae (icosahedral head and contractile tail). The phages retained higher lytic competence of 90.4; 92.68 and 95.25% for EAP1, EAP2 and EAP3, respectively. The phages were stable at strong alkaline (pH 10) 2% salt solution conditions and UV spectra. While EAP3 phage revealed the hexagonal head and very short tail that belongs to Myoviridae family. Bacteriophages were characterized by digestion of the phage DNA with three restriction endonucleases and were placed into three groups based on the patterns. Bacteriophages were 9 used for reducing bacterial infection populations and severity on pear. In a bioassay, the biocontrol of E. amylovora was evaluated using disks of immature pear fruit. On the pear disk surface, bacterial exudate was considerably suppressed by all phage isolates. According to measurements of the bacterial population still present on the disk surface, phage therapy could reduce it by up to 97%. Bacteriophages reduced pear fire blight disease severity on pear fruit trails.ConclusionThe results indicated that bacteriophage isolates may demonstrate variable reactivity against E. amylovora. Bacteriophages reduced pear fire blight disease severity on pear fruit trials. The results indicated that bacteriophage isolates may demonstrate variable reactivity against E. amylovora.

Analysis of natural resistance-associated macrophage protein-1 (NRAMP-1) level based on death, comorbidities and severity of COVID-19 patients: a cross-sectional study.

Determination of NRAMP-1 protein levels using the enzyme link-immunosorbent assay technique in death, had comorbidities and severity of clinical symptoms of COVID-19 patients. Of the 62 patients who received treatment, 10 patients died with an average NRAMP-1 level of 650ng/ml and 52 patients who survive with an average NRAMP-1 level of 1065.26ng/ml. The results of the study also found that 34 patients had comorbidities with an average NRAMP-1 level of 838.82ng/ml and 28 patients without comorbidities with an average NRAMP-1 level of 1191.92ng/ml. Based on the severity of clinical symptoms in survive patients, 10 patients with mild were found with an average NRAMP-1 level of 984.31ng/ml, with moderate in 31 patients with an average NRAMP-1 level of 1104.71ng/ml and severe in 11 patients with an average NRAMP-1 level of 1027.71ng/ml. NRAMP-1 protein levels were significantly lower in COVID-19 patients who died and had comorbidities.

Novel Monoclonal Antibodies Specific for Human Ki67 and P53 Tumor Markers in Breast Cancer Tissue Samples.

Ki67 and P53 are important diagnostic and prognostic biomarkers expressed in several cancers. The current standard method for evaluating Ki67 and P53 in cancer tissues is immunohistochemistry (IHC), and having highly sensitive monoclonal antibodies against these biomarkers is necessary for an accurate diagnosis in the IHC test. To generate and characterize novel monoclonal antibodies (mAbs) against human Ki67 and P53 antigens for IHC purposes. Ki67 and P53-specific mAbs were produced by the hybridoma method and screened by enzyme-linked immunosorbent assay (ELISA) and IHC techniques. Selected mAbs were characterized using Western blot and flow cytometry, and their affinities and isotypes were determined by ELISA. Moreover, using the IHC technique in 200 breast cancer tissue samples, we assessed the specificity, sensitivity, and accuracy of the produced mAbs. Two anti-Ki67 (2C2 and 2H1) and three anti-P53 mAbs (2A6, 2G4, and 1G10) showed strong reactivity to their target antigens in IHC. The selected mAbs were also able to recognize their targets by flow cytometry as well as Western blotting using human tumor cell lines expressing these antigens. The specificity, sensitivity, and accuracy calculated for clone 2H1 were 94.2%, 99.0%, and 96.6%, and for clone 2A6 were 97.3%, 98.1%, and 97.5%, respectively. Using these two monoclonal antibodies, we found a significant correlation between Ki67 and P53 overexpression and lymph node metastasis in patients with breast cancer. The present study showed that the novel anti-Ki67 and anti-P53 mAbs could recognize their respective antigens with high specificity and sensitivity and therefore can be used in prognostic studies.

Patchouli alcohol induces G0 /G1 cell cycle arrest and apoptosis in vincristine-resistant non-small cell lung cancer through ROS-mediated DNA damage.

Lung cancer, especially non-small cell lung cancer (NSCLC), is one of the leading causes of cancer-related deaths worldwide. Vincristine (VCR) is a chemotherapeutic agent for lung cancers; however, its effectiveness is limited by side effects and the development of drug resistance. Patchouli alcohol (PA), from Pogostemon cablin extract, is known to possess anti-inflammatory and anticancer properties. In this study, we investigated the role of PA in inducing reactive oxygen species (ROS)-mediated DNA damage in A549 and VCR-resistant A549/V16 NSCLC cells. The anticancer potential of PA was studied using the MTT assay, colony formation, flow cytometry analysis, western blotting, DCFDA staining, immunofluorescence staining, and TUNEL assay techniques. The intracellular ROS levels were enhanced in PA-treated cells, activating the CHK1 and CHK2 signaling pathways. PA further inhibited proliferation and colony-forming abilities and induced cell cycle arrest at the G0 /G1 phase by regulating p53/p21 and CDK2/cyclin E1 expression. Moreover, PA increased the percentage of cells in the subG1 phase and induced apoptosis by activating the Bax/caspase-9/caspase-3 intrinsic pathway. In addition, drug resistance (p-glycoprotein) and cancer stem cell (CD44 and CD133) markers were downregulated after PA treatment. Furthermore, combining PA and cisplatin exhibited synergistic inhibitory activity in A549 and A549/V16 cells. PA induced ROS-mediated DNA damage, triggered cell cycle arrest and apoptosis, attenuated drug resistance and cancer stem cell phenotypes, and synergistically inhibited proliferation in combination with cisplatin. These findings suggest that PA has the potential to be used for the treatment of NSCLC with or without VCR resistance.

Pressure-Driven Sample Flow through an Electrospun Membrane Increases the Analyte Adsorption

Electrospun polymer membranes are regarded as prospective biosensor components due to their large specific surface area and diverse opportunities for chemical modifications. However, their intricate porous structure can impede diffusion and render some analyte-binding sites inaccessible. To overcome these diffusion limitations and improve analyte adsorption onto the polymer, a pressure-driven sample flow through the membrane can be employed. To date, the efficiency of pressure-driven analyte delivery into these membranes has not been quantified. Here, we compare forced flow and passive sample diffusion through poly(dioxanone) electrospun membranes. We examine two model analytes, BSA and interleukin-1 beta (IL1b), to address both non-specific and specific binding. Following exposure of the membranes to the test solutions, we measured the residual concentrations of the analytes using fluorometry and enzyme-linked immunosorbent assay (ELISA) techniques. The pressure-driven sample loading was superior to passive diffusion, with a 2.8–11.5-fold change for physical adsorption and a 2.4–3.4-fold difference for specific binding. Our data can be useful for the development of immunoassays and microfluidic devices.

Biochemical and Yield Component of Hybrid Chili (Capsicum annuum L.) Resulting from Full Diallel Crosses

Chili (Capsicum annuum), economically important, is one of the world’s most popular horticultural plants. Functional biochemical components, such as polyphenol content, antioxidants, and α-glucosidase inhibitory properties, are found in chili. The purpose of this study was to evaluate a chili hybrid that resulted through full diallel crosses for its yield components, total phenolic (TPC) and flavonoid content (TFC), antioxidants, and α-glucosidase inhibitory (AGI) activities. The Folin-Ciocalteu and AlCl3-colorimetric assays were, respectively, used for TPC and TFC analyses. Using 2,2-diphenyl-1-picrylhydrazy (DPPH) and ferric reducing antioxidant power assay (FRAP) techniques, the antioxidant activity of a sample was determined. The bioassay of α-glucosidase inhibition was used to evaluate the antidiabetic activity of the sample. The twenty-five genotypes (hybrid and parent) have diverse yield components and biochemical contents. The highest fruit weight per plant was found in IPB074005 (1008.85 g). IPB114367 showed a high AGI (80.25%), antioxidant FRAP (43.42 µmol TE g−1 DW), TFC (3.97 mg QE g−1 DW), and TPC (37.51 mg GAE g−1 DW). These findings suggested that hybrid plants that suppress α-glycosidase and oxidative stress may prevent diabetes and its complications. This vital information could help design replacement drugs and diabetes diets.

Modulation of the Expression of Lipogenic Activity by Malted Hungry Rice Flour (Digitaria exilis.) on Rats Induced with Obesity

Lipolysis and lipogenesis causes obesity and these can be mediated by hormones. The study was aimed at determining the effect of malted hungry rice flour on lipogenic activity on diet-induced obese rats. The study was conducted between September to January 2021 and 2022. Twenty male adult albino rats were divided into four groups and obesity was induced which resemble mild obesity condition in human. The groups for the experiment were normal control (AIN-93), obese control, and two test diets were used to evaluate lipid profile parameter using a standard assay technique. There is an increase in BMI (from 0.37-0.41 to 0.58- 0.72). There are also an increase in other lipid profile with a decrease in HDL (p<0.05) after consumption of a palatable diet. Consumption of the test diet led to a significant reduction in the lipid profile parameter, and a significant increase in HDL cholesterol compared to the (group) obese control (p<0.05). The result suggests that malted hungry rice flour has an anti-obesity effect.

Hemocompatible Functionalized Hydrogen Substituted Graphdiyne Based Highly Durable Biosensor for Liver Cancer Detection.

Present work focuses on the development of a highly durable biosensor for liver cancer (LC) biomarker (Annexin A2; ANXA2) detection. In this work, we have modified hydrogen substituted graphdiyne (HsGDY) using an organofunctional silane [3-(aminopropyl)triethoxysilane (APTES)], leveraging the opposite surface polarities on HsGDY and APTES to fabricate a highly hemocompatible functionalized nanomaterial matrix. The high hemocompatibility of APTES functionalized HsGDY (APTES/HsGDY) allows long-term stabilized immobilization of antibodies in their native state, hence increasing the durability of the biosensor. The biosensor was fabricated using electrophoretic deposition (EPD) of APTES/HsGDY onto an indium tin oxide (ITO)-coated glass substrate at 40% lower DC potential than nonfunctionalized HsGDY with successive immobilization of monoclonal antibodies of ANXA2 (anti-ANXA2) and bovine serum albumin (BSA). The synthesized nanomaterials and fabricated electrodes were investigated using a zetasizer and spectroscopic, microscopic, and electrochemical (cyclic voltammetry and differential pulse voltammetry) techniques. The developed immunosensor (BSA/anti-ANXA2/APTES/HsGDY/ITO) could detect ANXA2 in a linear detection range from 100 fg mL-1 to 100 ng mL-1 with a lower detection limit of 100 fg mL-1. The biosensor demonstrated excellent storage stability of 63 days along with high accuracy toward detection of ANXA2 in serum samples of LC patients as validated via enzyme-linked immunosorbent assay technique.