Assay Method Research Articles

Fast screening of gold in rock samples using polyurethane foam extraction and inductively coupled plasma mass spectrometry determination.

In this work, the volume of sample solution and concentration of gold was optimized for extraction with foam in dimensions (1 × 1 × 1) and then was used for extraction from soil samples. The results showed that the proposed technique has a good analytical efficiency compared to the standard fire assay method and the accuracy of work is in the range of 74-125 %. The equation of linear calibration curve was obtained with regression coefficient better than 0.9997, and the detection and quantification limit of the gold in aqueous and soil sample obtained 0.25 and 0.8 µg kg -1, respectively.•This method is inexpensive and fast for determination of gold in various samples.•This method has high thought of sample determination.•This method is green chemistry method.

Huashi baidu granule alleviates inflammation and lung edema by suppressing the NLRP3/caspase-1/GSDMD-N pathway and promoting fluid clearance in a porcine reproductive and respiratory syndrome (PRRS) model.

Huashi Baidu Granule (HSBDG), a traditional Chinese medicine (TCM), is used for treating coronavirus disease 2019 (COVID-19). Porcine reproductive and respiratory syndrome (PRRS) is considered the "COVID-19" for swine. According to the TCM theory, "dampness" is the main pathogenic factor in COVID-19 and PRRS, and "Huashi" means that this formula is good at removing "dampness". Studies have demonstrated that HSBDG's effect in COVID-19; but the mechanism of removing "dampness" remains elusive. We aimed to assess the effect of HSBDG on PRRS, and elucidate its potential mechanism in removing "dampness". We established a PRRS-virus (PRRSV)-infected Marc-145cells model, and performed qRT-PCR, Western blot analysis, and indirect immunofluorescence assay to examine the anti-PRRSV effects of HSBDG in vitro. PRRSV-infected pig model was established and used to investigate HSBDG's effect in PRRS and explore underlying mechanisms in removing "dampness" using ELISA and immunohistochemistry assay methods. HSBDG exhibited anti-PRRSV activity and suppressed the viral replication and release phases. HSBDG treatment alleviated PRRS, lowered rectal temperature, reduced histopathological changes and viral load in lung tissues, and ameliorated organ lesions. Moreover, IL-1β, IL-6, IL-8, and TNF-α expressions were decreased in lung tissues. Mechanistically, HSBDG inhibited the NLRP3/Caspase-1/GSDMD-N pathway to reduce the inflammatory response and upregulated AQP1, AQP5, α-ENaC, and Na-K-ATPase expressions to promote lung fluid clearance. HSBDG exerted anti-PRRSV effects and could attenuate PRRS. HSBDG potentially removes "dampness" by attenuating inflammation by suppressing the NLRP3/Caspase-1/GSDMD-N pathway and inhibiting pulmonary edema by upregulating the expression of AQP1, AQP5, α-ENaC, and Na-K-ATPase.

Postoperative evaluation of interleukin-8 and C1q/TNF-related protein-12 in patients undergoing coronary artery bypass graft surgery.

Coronary artery bypass graft surgery is one of the most frequently performed surgeries worldwide. Coronary artery bypass graft surgery induces an inflammatory response. Interleukin-8 is a pro-inflammatory cytokine that plays a role in the pathogenesis of cardiovascular diseases. C1q/TNF-related protein-12 is implicated in mitigating inflammation and cardiomyocyte damage. This study aimed to compare interleukin-8 and C1q/TNF-related protein-12 levels before and 45 days after coronary artery bypass graft surgery. A total of 43 patients who underwent coronary artery bypass graft surgery were studied. Serum concentrations of interleukin-8 and C1q/TNF-related protein-12 were evaluated using the enzyme-linked immunosorbent assay method before and 45 days after the surgery. No significant differences were observed in interleukin-8 levels between pre- and post-coronary artery bypass graft surgery (p=0.077). However, serum levels of C1q/TNF-related protein-12 were found to be lower 45 days after coronary artery bypass graft surgery compared to pre-surgery levels (p<0.001). Moreover, changes in C1q/TNF-related protein-12 were not associated with diabetes, hypertension, and body mass index (p>0.05), but C1q/TNF-related protein-12 alterations were found to be associated with opium addiction. These findings suggest that the evaluation of C1q/TNF-related protein-12 can be beneficial for the late assessment of post-coronary artery bypass graft surgery inflammation. The reduction of C1q/TNF-related protein-12 levels might indicate increased levels of inflammation after surgery at this time point, which requires the assessment of further inflammatory factors to confirm this finding.

Investigating the Role of Quercetin, an Active Ingredient in Bazhen Decoction, in Targeting CXCL8 to Inhibit Macrophage M2 Polarization and Reshape the Immunological Microenvironment of Colorectal Cancer.

Bazhen Decoction (Eight Treasures Decoction) has demonstrated efficacy in the treatment of colorectal cancer (CRC), yet the active ingredients in it and the mechanisms underlying their anti-cancer properties are not well understood. Through network pharmacology, the effective components of Bazhen Decoction against CRC and their corresponding key genes were delineated. Molecular docking was executed to identify the active component targeting the key gene CXCL8, which led to the discovery of Quercetin. The cellular thermal shift assay method was then used to verify the binding interaction. CRC cells were treated with incremental concentrations of Quercetin, cell viability was evaluated by the Cell Counting Kit-8 assay to calculate the IC50, and apoptosis rates were determined by flow cytometry. Expression of the apoptosis-related proteins Bcl-2 and Cleaved caspase-3 was measured using western blot. The impact of Quercetin on macrophage polarization was studied by co-culturing the treated CRC cells with macrophages, assessing M1 and M2 macrophage distribution via flow cytometry, and quantifying cytokine levels (IL-6, IL-10, IL-12, and CXCL8) with enzyme-linked immunosorbent assay (ELISA). The active ingredient Quercetin from Bazhen Decoction exhibited a targeted binding affinity with the key gene CXCL8, which enabled it to inhibit the proliferation of CRC cells and induce cell apoptosis. The overexpression of CXCL8 was associated with the promotion of CRC malignancy, yet the presence of Quercetin could lessen the impact of CXCL8 overexpression on CRC cells. Moreover, the treatment with Quercetin leads to a diminished abundance of M2 macrophages and an increase in the levels of cytokines IL-6 and IL-12, while reducing the levels of IL-10 and CXCL8, which indicates that Quercetin has an inhibitory effect on macrophage M2 polarization. Quercetin, the active component in Bazhen Decoction that is known for anti-CRC effects, targets and inhibits CXCL8 to impede the malignant behaviors and the M2 polarization of macrophages. Thus, Quercetin may be utilized as an immunomodulatory agent in CRC treatment.

Skeletal muscle ribosome analysis: A comparison of common assay methods and utilization of a novel RiboAb antibody cocktail.

While total RNA concentrations putatively represent ribosome content, there is a need to homologize various quantification approaches. Thus, total RNA concentrations ([RNA]) provided through UV-Vis spectroscopy (UV), fluorometry-only (Fluor), and fluorometry-based microfluidic chip electrophoresis (MFGE) were examined in C2C12 myotubes and mouse skeletal muscle to determine if values aligned with [18S + 28S rRNA] (i.e., criterion ribosome metric). A novel antibody cocktail (termed RiboAb) was also tested and compared to [18S + 28S rRNA] in these models. In myotubes, 24-h IGF-1 treatments increased [18S + 28S rRNA] (~2.0-fold) and [RNA] based on UV (~1.9-fold), Fluor (~2.3 fold), and MFGE (~2.1-fold). In C57BL/6 mice, 10 days of mechanical overload (MOV) elevated plantaris [18S + 28S rRNA] (~1.7-fold) and [RNA] according to UV (~1.5-fold), Fluor (~1.6-fold), and MFGE (~1.8-fold). Myotube and mouse plantaris RiboAb levels were significantly higher with IGF-1 treatments and MOV, respectively, versus controls (1.3-fold and 1.7-fold, respectively), and values correlated with [18S + 28S rRNA] (r = 0.637 and r = 0.853, respectively, p ≤ 0.005). UV, Fluor, and MFGE [RNA] are seemingly valid surrogates of cell/tissue ribosome content, although each method has advantages (e.g., ease of use) and disadvantages (e.g., magnitudes of bias) discussed herein. Finally, the RiboAb cocktail may also represent ribosome content, although this should be further explored in other models.

Effect of long-term negative energy on appetite hormone levels in individuals with prediabetes and diabetes.

Calorie restriction and exercise are commonly used first interventions to prevent the progression of prediabetes and alleviate the symptoms of type 2 diabetes. Our study was designed to determine the effect of the energy deficit caused by long-term (12-week) calorie restriction and exercise programs on appetite responses in obese individuals with prediabetes and type 2 diabetes. Calorie restriction and exercise programs appropriate for age, gender, and work environment were applied to 22 individuals with prediabetes and 22 with type 2 diabetes participating in the study for a period of 12 weeks. Ghrelin, glucagon-like peptide-1, and peptide tyrosine tyrosine values of samples taken before and after treatment were determined by the enzyme-linked ιmmunosorbent assay method. Appetite hormone levels did not change after calorie restriction and exercise in the prediabetes group (p>0.05). In the diabetes group, calorie restriction and exercise significantly increased ghrelin and peptide tyrosine tyrosine concentrations (p<0.005). Additionally, when all patients were evaluated together, ghrelin, glucagon-like peptide-1 and peptide tyrosine tyrosine levels differed significantly after the intervention (p<0.005). The energy deficit created by long-term calorie restriction and exercise did not modulate the appetite hormones in prediabetic and obese individuals. However, increased ghrelin and peptide tyrosine tyrosine levels in individuals with diabetes support that the same treatment program is an effective method to regulate appetite hormones.

Comparison of Leptin and Tnf-Α Levels in Type 2 Diabetes Mellitus Patients with and Without Obesity

Background: Leptin and tumor necrosis factor alpha (TNF-α) are the main adipokine products of adipose tissue, leptin is a hormone that controls appetite, while TNF-α acts as a proinflammatory cytokine that can cause Insulin resistance Leptin and TNF-α levels are influenced by body mass inde (BMI). Objective: To prove the differences in leptin and TNF-α levels between T2DM patients with and without obesity. Material and Method: A cross-sectional study was conducted on 31 T2DM subjects with obesity and 29 DMT2 subjects without obesity. Leptin and TNF-α levels were measured using the enzyme linked immunosorbent assay (ELISA) method. Obesity status was determined based on BMI values where BMI ≥25 kg / m2 was the obesity group and BMI 25 kg / m2 group without obesity. The t-test was used as the comparative test. Results: The median leptin levels in the obese and non-obese T2DM groups were 31.8(7;74.6) ng/mL and 9.2(0.8;36.7) ng/mL. There was a difference in leptin levels between the two groups with p=0.001. The median TNF-α levels in the obese and non-obese DMT2 group were 6.6(2.2;21.2) pg/mL, and 4.4(1.6;15.8) pg/mL. There was a difference in TNF-α levels between the two groups with p=0.007. Conclusion: There are significant differences in leptin and TNF-α levels between T2DM patients with and without obesity, where leptin and TNF-α levels are higher in T2DM patients with obesity than T2DM patients without obesity.

Dynamics of cortisol indicators in patients with mild arterial hypertension I-II stages in the conditions of a stress test against the background of excessive anxiety and hyperventilation syndrome

Annotation. The aim of the study was to investigate the dynamics of cortisol levels in 105 patients with mild (grade 1) arterial hypertension (AH) of stage I-II (st.) against the hyperventilation syndrome (HVS) and excessive anxiety, in the conditions of a 6-minute walk test (6MWT). The control group of patients included 28 patients with mild hypertension, I-II stages, without signs of vegetosomatic disorders. The state of anxiety and HVS was determined by the Spielberger-Hanin scale and the Nijmegen questionnaire. The level of cortisol in saliva was determined by enzyme-linked immunosorbent assay method on the “Stat Fax 303 Plus” analyzer. Statistical analysis was performed using the Statistica v.10 program (USA). It was found that normal baseline cortisol levels in patients of the main group with AH I stage were found in 36.96% of patients and amounted to 5.63±0.33 nmol/l, in AH II stage - in 41.46% of patients. The conditional norm was the result obtained in the group of practically healthy individuals. In the control group with I-II stage of hypertension, normal baseline values were observed in 65.5% and 58.33% of patients, respectively. Increased baseline cortisol levels (6.58±0.32) nmol/l were detected in 47.83% of patients in the main group with AH I stage, and in 46.34% of patients with AH II stage (6.72±0.26) nmol/l (p˂0.05); in the control group, respectively, in 37.5% and 41.67% of patients. The rest of the patients, reduced baseline cortisol levels were found. After 6 MWT, in the main group with stage I hypertension, the number of cases with elevated cortisol levels increased by 8.70%, and the number of patients with low levels decreased by 10.87%. In stage II hypertension, after 6 MWT, the number of patients with elevated cortisol levels increased by 12.20%, while the number of cases with low levels decreased by 7.32% and the number of normal levels decreased by 4.88%. In the control group with stage I hypertension after 6 MWT, a decrease in normal cortisol levels was noted in 6.25% of patients and a similar increase in the number of patients with elevated levels. In stage II hypertension after 6 MWT, cases with normal cortisol levels decreased by 16.33%; at the same time, the number of patients with elevated levels increased by 8.33%. The dynamics of cortisol levels after exercise in mild hypertension with excessive anxiety and hypertension indicates the tension of neuroendocrine regulation mechanisms, the risk of destabilization of the underlying disease under stress.

The Association of Serum Lactoferrin Level with Psychological Symptoms, Cognition, and Executive Function in Schizophrenia Patients

Objective: Lactoferrin, a glycoprotein, has known neuroprotective effects, yet its role in the pathophysiology of neuropsychiatric disorders, particularly schizophrenia, remains unclear. This study aims to assess changes in lactoferrin levels during different phases of schizophrenia and explore its relationship with cognitive symptoms and performance. Method: This before/after interventional study involved 30 patients diagnosed with schizophrenia. Participants were evaluated at two time points: upon hospital admission and after the resolution of acute symptoms. The Positive and Negative Syndrome Scale (PANSS) was utilized to measure symptom severity, while the Brief Assessment of Cognition in Schizophrenia (BACS) assessed the neurocognitive function. Serum lactoferrin levels were quantified using the Enzyme-Linked Immunosorbent Assay (ELISA) method. Results: Serum lactoferrin levels significantly decreased from 130.63 ± 52.49 ng/mL at admission to 85.42 ± 29.03 ng/mL at discharge (P &lt; 0.001). No significant correlation was found between lactoferrin levels and PANSS scores (r = 0.011, P = 0.975). However, an inverse correlation was observed between changes in lactoferrin levels and the executive function subscale of the BACS (r = -0.360, P = 0.050). Cognitive assessments indicated significant improvements in verbal memory (P = 0.033), working memory (P = 0.002), and executive function (P = 0.039) post-treatment. Conclusion: The study demonstrates a significant reduction in serum lactoferrin levels during the acute phase of schizophrenia, suggesting its potential role in modulating cognitive functions, particularly the executive function, rather than influencing positive or negative symptoms.

Diagnostic Accuracy of Enzyme-Linked Immunosorbent Assay Compared with Polymerase Chain Reaction in Diagnosing &lt;i&gt;Helicobacter pylori&lt;/i&gt; in Patients with Gastric Problems

Background: Helicobacter pylori, identified in 1982, is a significant gastric pathogen affecting a large portion of the global population. Objectives: This study aims to compare the diagnostic accuracy of enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) methods for detecting H. pylori, which is crucial for improving treatment strategies for gastric disorders. Methods: The study involved 70 patients at Khurshid Private Laboratory in Tehran between July and September 2022. The goal was to identify H. pylori DNA in stool samples using real-time PCR, while simultaneously testing serum samples for H. pylori using the ELISA method. Data were analyzed using SPSS software. Results: In a study of 70 patients with stomach diseases, the prevalence of H. pylori infection based on PCR results was 24.29%, with a slightly higher prevalence in females than males. For diagnosing H. pylori infection, the optimal IgA cut-off point was identified as 7.15 mg/dL, yielding a sensitivity of 88%, specificity of 89%, and an AUC of 0.94. The optimal cut-off for IgG was 95.70 mg/dL, with a sensitivity of 76%, specificity of 94%, and an AUC of 0.88, using PCR results as the gold standard. Conclusions: This study emphasizes the importance of accurately diagnosing H. pylori infections, highlighting that PCR offers greater sensitivity and specificity than ELISA. By establishing optimal IgA and IgG cut-off points, the study enhances diagnostic precision, which could lead to improved therapeutic interventions and patient outcomes.

Antimicrobial resistance profile, biofilm forming capacity and associated factors of multidrug resistance in Pseudomonas aeruginosa among patients admitted at Tikur Anbessa Specialized Hospital and Yekatit 12 Hospital Medical College in Addis Ababa, Ethiopia

BackgroundPseudomonas aeruginosa is one of the leading causes of nosocomial infections and the most common multidrug-resistant pathogen. This study aimed to determine antimicrobial resistance patterns, biofilm-forming capacity, and associated factors of multidrug resistance in P. aeruginosa isolates at two hospitals in Addis Ababa, Ethiopia.MethodsA cross-sectional study was conducted from August 2022 to August 2023 at Tikur Anbessa Specialized Hospital and Yekatit 12 Hospital Medical College. Culture and identification of P. aeruginosa were done using standard microbiological methods. An antimicrobial susceptibility test was done by Kirby-Bauer disk diffusion according to CLSI recommendations. The microtiter plate assay method was used to determine biofilm-forming capacity. SPSS version 25 was used for data analysis. Bivariate and multivariable logistic regression were used to assess factors associated with multidrug resistance in P. aeruginosa. The Spearman correlation coefficient (rs = 0.266)) was performed to evaluate the relationship between biofilm formation and drug resistance.ResultsThe overall prevalence of P. aeruginosa was 19.6%. High levels of resistance were observed for ciprofloxacin (51.8%), ceftazidime (50.6%), and cefepime (48.2%). The level of multidrug-resistance was 56.6%. The isolates showed better susceptibility to ceftazidime-avibactam (95.2%) and imipenem (79.5%). Overall, 95.2% of P. aeruginosa were biofilm-producing isolates, and 27.7% and 39.8% of isolates were strong and moderate biofilm producers, respectively. A positive correlation and statistically significant relationship was observed between resistance to multiple drugs and the level of biofilm formation (rs = 0.266; p-value = 0.015). Previous history of exposure to ciprofloxacin (OR, 5.1; CI, 1.12–24.7, p-value, 0.032) was identified as an independent associated factor for multidrug resistance in P. aeruginosa.ConclusionThe present study indicates an association between multidrug resistance in P. aeruginosa and its biofilm formation capabilities. Additionally, over half of the isolates were resistant to multiple drugs, with prior use of ciprofloxacin linked to the development of multidrug-resistance. These findings suggest that antibiotic stewardship programs in hospital settings may be beneficial in addressing resistance.

A Novel Stability Indicating Analytical Development and Validation of an RP-HPLC Assay Method for the Quantification of Voglibose in Bulk and its Formulation

The investigation developed a new stability-indicating Reverse phase high-pressure liquid chromatographic (RP-HPLC) assay method to quantify Voglibose in bulk and pharmaceutical dosage form. The technique used isocratic elution on an Amino (NH2) column (250x4.6mm, 5µ particle size) with a mobile phase of Phosphate Buffer (pH 6.7) and Acetonitrile (27:73 v/v). The separation was monitored using UV detection at 210 nm. The method was validated, meeting ICH guidelines. The retention time of Voglibose was 6.38 minutes, and the linearity was between 24-56 µg/ml. The limit of detection and quantitation were 0.70 and 2.11 µg/ml, respectively. Analytical performance parameters were determined according to ICH guidelines. The method was simple, precise, accurate, robust, and rapid, making it suitable for quantifying Voglibose in bulk and tablet formulation.

Combined role of PEAR1 and GP genes in the mechanism of aspirin resistance and its clinical significance

Exploring the mechanism of aspirin resistance helps to improve the prevention and treatment of ischemic stroke diseases and reduce the interference of resistance mechanism on the therapeutic effect of aspirin. In this paper, the Affiliated Hospital of Beihua university, together with two other hospitals, was chosen as the research unit, from which 260 AIS patients were selected for aspirin resistance study. The joint role of PEAR1 gene and GP gene in the mechanism of aspirin resistance was discussed by genotyping, TEG assay, univariate analysis, and binary logistic analysis methods. Among the GP1BA genes in AIS patients, the frequency of CC-type genes was as high as 92.03%, and the risk of aspirin resistance was significantly increased in CC-type relative to TT-type. The incidence of rs5924 allele T was significantly higher in the AR group than in the AS group (OR = 1.632, p = 0.006), and there was a significant effect of diabetic conditions on aspirin resistance at the 1% level. Diabetes significantly inhibits plasma proteins in AIS patients and makes aspirin less inhibitory to platelets. Therefore, clarifying the influence of PEAR1 and GP genes in the mechanism of aspirin resistance can be used to obtain the precise loci from the gene therapy point of view to enhance the disease treatment effect of aspirin.

Metabolite profiling, antioxidant, and in vitro wound healing activities of Citrus medica L. and Citrus x microcarpa Bunge peels and leaves essential oils

Wound care is still dominated by the use of synthetic chemicals such as anti-inflammatory drugs, corticosteroids and antibiotics which have many shortcomings. One potential alternative to be developed is essential oils. Citrus are members of the Rutaceae family which contain essential oils. Citrus medica L. and Citrus x microcarpa Bunge are distributed in West Sumatra and research on wound healing activity is still lacking. Therefore, this study aims to identify the metabolite profiles of essential oils of two species of citrus using FTIR spectroscopy and GC-MS as well as evaluate their antioxidant activity and determine their wound healing activity. Essential oils were obtained by the hydrodistillation method. Metabolite fingerprinting and profiling were carried out by using FTIR and GC-MS methods. Antioxidant activity was determined using the ABTS and FRAP methods while wound healing used the MTT Assay and Scratch Assay. The result showed the secondary metabolites of Citrus medica peels (LPEO) and Citrus medica leaves (LLEO) essential oil contained D-limonene and citral. Meanwhile Citrus x microcarpa leaves (MLEO) contain β-pinene, germacrene D, and elemol and Citrus x microcarpa peels essential oil (MPEO) consists of D-limonene, β-pinene, and germacrene D. MLEO has stronger antioxidant activity than MPEO, LPEO and LPEO with ABTS and FRAP method with IC50 197.051 µg/mL and 8.04 Fe (II)/mg sample and is followed by LLEO. Testing of wound healing activity using MTT assay showed that MPEO significantly increased cell proliferation. Meanwhile, the highest wound closure was found at a concentration of 0.1 µg/mL for MLEO followed by LPEO at 77.54% and 73.71% using scratch assay method. It can be concluded that essential oils of Citrus medica and Citrus x microcarpa can increase cell proliferation and migration so that they have potential to be developed as an alternative in wound healing.

Application of Fish Embryo Assay Using Zebrafish and Oryzias latipes for Toxicity Testing and Deriving Water Quality Criteria

To protect aquatic organisms in ecosystems, each country and continental union has established guidelines for deriving the water quality standards (WQS) of specific substances. These guidelines mandate the use of acute and chronic toxicity data for fish, which are high-trophic-level organisms. However, due to increasing concerns about animal welfare and experimental ethics, there is a growing need for alternative methods to determine substance toxicity in fish. Fish toxicity tests using early life stages, such as embryos or larvae, have been utilized as alternative methods for adult fish toxicity assays. This review of the WQS guidelines and relevant test protocols confirmed the classification of acute and chronic toxicity in fish assays using different developmental stages. Fish toxicity data derived from exposure periods longer than one week using embryonic- or larval-stage organisms can be considered as indicative of chronic toxicity. There is a high correlation between fish embryo toxicity and adult effects, suggesting that fish embryo toxicity tests with appropriate exposure durations could replace adult fish toxicity tests, addressing experimental animal ethics concerns.

Lower Plasma β-Amyloid 1-42 Levels in Amnestic Mild Cognitive Impairment Compared to Healthy Individuals

BACKGROUND: Amnestic mild cognitive impairment (aMCI) is strongly associated with an increased risk of progression to Alzheimer’s disease (AD). In AD, cerebrospinal fluid (CSF) β-Amyloid 1-42 levels are known to decrease, a pattern which is also observed in aMCI. While in AD, apolipoprotein E (ApoE) ε4 allele is known to be a genetic risk factor, the role of ApoE ε4 allele in modulating plasma β-Amyloid 1-42 levels in aMCI remains unclear. Therefore, this study was performed to evaluate plasma β-Amyloid 1-42 levels in aMCI patients compared to cognitively healthy individuals and investigate its association with ApoE ε4 allele.METHODS: A cross-sectional study involving 57 aMCI and 54 cognitively healthy control (HC) subjects was performed. Blood samples were taken from subjects from both groups for measurement of the plasma β-Amyloid 1-42 and ApoE ε4 allele. The plasma levels of β-Amyloid 1-42 were measured using an enzyme-linked immunosorbent assay (ELISA) methods, while the ApoE ε4 allele genotyping was conducted using polymerase chain reaction (PCR) techniques.RESULTS: Plasma β-Amyloid 1-42 in individuals with aMCI (23.9 pg/mL) was significantly lower than that in HC (25.3 pg/mL) with cut-off value of 24.6 pg/mL (AUC: 70.8%; 95% CI: 61.1–80.5%; p&lt;0.001) sensitivity of 64.8%, and specificity of 71.9%. There was no significant association between plasma β-Amyloid 1-42 and the ApoE ε4 allele. However, plasma β-Amyloid 1-42 in ε4 carriers were lower than in ε4 non-carriers.CONCLUSION: Lower plasma β-Amyloid 1-42 levels were observed in aMCI patients compared to cognitively healthy individuals, suggesting its potential as a biomarker for identifying aMCI.KEYWORDS: blood biomarkers, amyloid beta peptides, amnestic mild cognitive impairment (aMCI)

Chrydintins A and B, two new sesquiterpenoids from Chrysanthemum indicum with cardioprotective activities.

Chrydintins A (1) and B (2), two new sesquiterpenoids were isolated from Chrysanthemum indicum., together with four known ones identified as 9-hydroxyeudesma-4,7(11)-dien-6-one (3), (7S,9S,10S)-9-hydroxy-11-methoxyeudesm-4-en-6-one (4), curcumenone (5), and polydactin B (6), respectively. The structures of isolated compounds were determined by HR-ESI-MS, NMR analysis, and quantum chemical calculation. The in vitro cardioprotective activities of all compounds were studied on H2O2-injured H9C2 cells using MTT assay method. Compound 5 exhibited considerable protective effect with cell viability at 81.45 ± 1.07%.

In Vitro Assessment of Antimicrobial Activity and Synergistic Effects of Ethanolic Extracts from Six Medicinal Plants

This in vitro experimental study examines and evaluates the antimicrobial and synergistic effects of the ethanolic extract of six plants: Biancaea sappan (L.) Tod, Bauhinia malabarica Roxb, Carthamus tinctorius L., Derris scandens (Roxb.) Benth, Hibiscus sabdariffa L., and Piper nigrum L. against common microbial species representing gram-positive, gram-negative bacteria, and fungi, consisting of Staphylococcus aureus, Escherichia coli, Staphylococcus epidermidis, Pseudomonas aeruginosa, and Candida albicans. The plants were extracted using 90% ethanol. According to the standard method of agar diffusion assay, the micro-dilution method for minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined. This study found that among the six plants, only B. sappan and B. malabarica exhibited moderate inhibitory effects against S. aureus and S. epidermidis. B. sappan had MIC values of 250 µg/ mL and 125 µg/ mL, respectively, and B. malabarica showed MIC values of 62.50 µg/ mL and 31.25 µg/ mL, respectively. The synergistic effects of a combination of B. sappan and B. malabarica extracts at a ratio of 25:75 were analyzed, and it was found that the combination inhibited S. aureus and S. epidermidis with MIC values of 250 µg/ mL and 125 µg/ mL, respectively. The fractional inhibitory concentration index (FICI) and the fractional bactericidal concentration index (FBCI) indicated antagonistic or synergistic effects of the combination, with FICI and FBCI values of 2.5–5.0 for both B. sappan and B. malabarica extracts in the 25:75 mixture. In conclusion, single plant ethanolic extracts of B. sappan and B. malabarica possess potent antimicrobial activity to varying degrees. However, the antimicrobial potency of the 25:75 ratio mixture of these extracts was shown to decrease against the same organisms, with in vitro antimicrobial activity and antagonistic effects observed only against the tested gram-positive bacteria.

Prevalence of non-tuberculous mycobacteria among people with acid-fast positive presumptive tuberculosis in Mali.

Non-tuberculous mycobacteria (NTM) are environmental agents that can cause opportunistic pulmonary disease in humans and animals, often misdiagnosed as tuberculosis (TB). In this study, we describe the cases of NTM identified during the first national anti-TB drug resistance survey conducted in Mali and explore associated risk factors. Sputum was collected from people presenting for pulmonary TB diagnosis from April to December 2019, regardless of age. Microscopy-positive patients were enrolled and tested using the Xpert MTB/RIF assay. A patient who tested negative for the Mycobacterium tuberculosis complex (MTBC) was tested for the presence of mycobacteria by amplification of the IS6110 and 16SrRNA (16S) genes through double quantitative real-time PCR, followed by nested PCR and Sanger sequencing of the IS6110-negative samples for NTM species identification. 1,418 sputum smear-positive patients were enrolled, including 1,199 new cases, 211 previously treated cases, and 8 whose previous treatment history was unknown. Based on the results of Xpert MTB/RIF assay and in-house PCR methods, 1,331 (93.9%) patients were positive for MTBC, 48 (3.4%) for NTM, and no species identification was possible for 39 (2.7%). Advanced age of 65 and over with an OR 8.8 (95% CI 2.3-33.2 and p = 0.001) and previous TB treatment with an OR 3.4 (95% CI 1.2-9.6 and p = 0.016) were the risk factors statistically associated with NTM detection. M. avium complex (MAC) was the predominant NTM species, detected in 20 cases. Detection of NTM in people presumed to have TB is an ongoing challenge, confounding correct TB diagnosis. Concomitant use of microscopy and GeneXpert testing among at-risk individuals could improve patient management.

Validation study on the assay method for anti-factor IIa potency of enoxaparin sodium.

Enoxaparin sodium is a low molecular mass heparin essential for effective anticoagulation therapy. However, significant variations in testing methods across different manufacturers have led to poor reproducibility of results, increasing the risks associated with drug quality evaluation by manufacturers and regulatory oversight. This study integrates the strengths of various testing methods to establish a reproducible assay that has been thoroughly validated. The validation results demonstrate that the method exhibits excellent specificity, linearity, robustness, precision, and accuracy, with recovery rates ranging from 98.0% to 102.0%. The new method demonstrated high consistency and reproducibility, with an RSD value of less than 2.0%, and showed the potential to replace the European Pharmacopoeia method by reducing reagent usage, experimental costs, and equipment requirements. The reliable results of this method facilitate its adoption across different laboratories, enhance the quality control of enoxaparin sodium, and provide a reference for new manufacturers and drug regulatory authorities, thereby ensuring medication safety.