Chloride Assay Research Articles

Evaluation of Antimicrobial Performance of Calcium Dihydroxide (Ca(OH)2) Coating on Ti for Potential Metallic Orthopedic Implant Applications

Background/Objectives: Orthopedic implant infections are rare but represent a significant problem for patients, surgeons, and the healthcare systems. This is because these infections cause severe and persistent pain and, in some cases, may require revision of the implant, among other things. Thus, there is strong interest in the use of antimicrobial coatings on orthopedic implants. Here, we investigate electrochemically deposited Ca(OH)2 antimicrobial coating for its potential to be used on metallic orthopedic implants. Methods: A triphenyl tetrazolim chloride (TTC) assay and isothermal microcalorimetry (IMC) were used to determine the reduction in microbial activity on three sets of Ti parts (discs and screws): uncoated, coated with hydroxyapatite (HA), and coated with Ca(OH)2. Results: Using the TTC assay, a ~70% reduction in the growth of bacteria on Ca(OH)2-coated discs was found, and using IMC, bacterial growth on these discs showed a decreased rate and an increased lag phase up to 25 h. Each of these sets of results was statistically superior to the corresponding results obtained using the other sets of parts. Conclusions: The present results suggest that the Ca(OH)2 coating may have potential for use on metallic orthopedic implants.

Some Biological Activities of Ethanol Extract of Aerial Parts of Gentiana olivieri

IIn recent years, due to the possible side effects of synthetic drugs, people have turned to natural drugs in combating diseases. Plants are important natural resources with these properties. In our study, some biological activities of the aerial parts of Gentiana olivieri were determined. In this context, the plant was extracted with ethanol in a Soxhlet apparatus. The antioxidant activity of the plant was measured with Rel Assay TAS and TOS kits. Its antimicrobial activity was determined by the agar dilution method. Determination of total phenolic content was determined using the Folin-Ciocalteu reagent. Total flavonoid quantification was performed using aluminum chloride assay. As a result of the analyzes, the TAS value of the ethanol extract of the aerial parts of Gentiana olivieri was determined as 7.775±0.114 mmol/L, the TOS value as 12.252±0.094 μmol/L and the OSI value as 0.158±0.002. In addition, total phenolic content was measured as 104.92±1.40 mg/g, total flavonoid content as 73.83±1.29 mg/g. In addition, it was determined that the plant extract was effective against standard bacterial and fungal strains at extract concentrations in the range of 50-200 μg/mL. As a result, it was determined that Gentiana olivieri could be an important natural source in terms of antioxidant and antimicrobial activity.

In Vitro Antioxidant Activities and Bioactive Compound Profiles of Extracts from the Brown Alga Ecklonia cava

Purpose: Ecklonia cava is an edible brown alga with documented antitumor, anti-inflammatory, and antioxidant activities. This study analyzed the free radical scavenging capacities and bioactive compound profiles of E. cava extracts to provide foundational evidence for potential uses as a functional raw material in various food, health, and beauty products.Methods: The free radical scavenging activities of ethanol and distilled water extracts of E. cava were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assays. The total phenolic content was determined quantitatively using the Folin-Ciocalteu reagent, with gallic acid as the standard. And also, the flavonoid content was determined using aluminum chloride assay, with quercetin as the standard.Results: Extracts prepared using 100% ethanol (100 E), 50% ethanol (50 E), and distilled water (DW) all demonstrated substantial dose-dependent free radical scavenging activities, with the 100 E extract showing the highest scavenging capacity according to both DPPH and ABTS assays, followed by 50 E and DW extracts. Moreover, the 100 E extract exhibited scavenging activity roughly equivalent to the positive control ascorbic acid. The 100 E extract also possessed the highest total polyphenol and flavonoid contents.Conclusion:Ecklonia cava holds promise as a functional material for health foods, cosmetics, and natural therapeutics.

Production of phenolic compounds, flavonoids, and lupeol by Lycium schweinfurthii suspension cultures in shake flasks and a rocking-motion bioreactor

Lycium schweinfurthii, a wild shrub of the Solanaceae family, has received increasing attention in the last decade for its therapeutic potential in traditional medicine due to its diverse array of secondary metabolites, including phenolic substances and terpenoids. The aim of this study was to investigate the accumulation of phenolics, flavonoids, and the terpenoid lupeol in L. schweinfurthii cell suspension shake flask cultures and a single-use 2-dimensional rocking motion bioreactor. Three different media formulations were compared for in vitro cell cultures. Various parameters, such as biomass accumulation, settled cell volume, cell viability (assessed via a 2,3,5-triphenyl tetrazolium chloride assay), and sucrose consumption were determined as indicators of cell activity and growth. Total phenolic and flavonoid contents were estimated spectrophotometrically, lupeol was quantified via High-Performance Thin Layer Chromatography (HPTLC). Although a higher fresh biomass concentration of 464 g L− 1 was obtained in MS medium supplemented with a combination of each, 1 mg L− 1 of 2,4-Dichlorophenoxyacetic acid (2,4-D) and 1-Naphthaleneacetic acid (NAA), the rocking-motion bioreactor cultivation was performed with 2 mg L− 1 NAA due to its superior reproducibility in viability, productivity, and content of bioactive compounds (e.g., phenolics, flavonoids, lupeol). A final fresh biomass concentration of 185 g L− 1 was achieved in a 16 L cultivation scale with a notable increase in the concentration of phenolics (1.4-fold) and flavonoids (1.7-fold). Most importantly, the concentration of lupeol, a pentacyclic triterpenoid known for its anti-inflammatory, antibacterial, and anti-atherogenic properties, exhibited a remarkable 5.5-fold increase in the bioreactor cultivation (585 µg g− 1) compared to shake flask cultivations (106 µg g− 1). The current study demonstrated the profound impact of media composition and non-limited fed-batch conditions in a rocking-motion bioreactor on the accumulation of bioactive compounds. The findings are also relevant to other plant cell cultures.

Comprehensive analysis of antioxidant and antibacterial activities of water and methanol extracts of Hibiscus flower

This study provides a comprehensive investigation into the antioxidant and antibacterial properties of Hibiscus flower extracts, highlighting their potential for future applications in functional foods. The research addressed key challenges by employing a multi-faceted approach, including four distinct methods for evaluating antioxidant potential: DPPH and FRAP assays, reducing power, and chelating ability of Fe ions. Total phenolic content was quantified using the Folin–Ciocalteu method, while tannins and flavonoids were assessed via vanillin–HCl and aluminum chloride assays. Additional analyses included spectrophotometric measurement of total flavonols and anthocyanins. Antibacterial activity was determined using a modified agar disk diffusion method, with results indicating significant inhibition of Salmonella typhimurium and Staphylococcus aureus by both aqueous and methanolic extracts. Colorimetric analysis revealed a reduction in chroma and hue angles, while FTIR spectroscopy identified key functional groups, such as suberin, lipids, and polysaccharides. Notably, the antioxidant efficacy varied depending on the solvent used, with aqueous extracts showing higher total phenolic content (4325.12 mg GAE/100 g) compared to methanolic extracts (3487.05 mg GAE/100 g). The study concludes that Hibiscus flower extracts, rich in antioxidants like tannins and anthocyanins, possess significant potential for use as natural preservatives and colorants in food products, supporting their role in developing novel functional foods.

Investigation of the Anticholinesterase and Antioxidant Activity of Several Iranian Thymus caramanicus Populations

Background: Thymus extracts have become attractive targets for screening bioactive compounds with potential applications in pharmaceuticals, food, and cosmetics industries. Objectives: This is the first report assessing the anticholinesterase and antioxidant activities, along with total phenolic and flavonoid contents (TPC and TFC), of hydroethanolic extracts from six populations of Thymus caramanicus Jalas at vegetative and flowering stages, collected from different regions in Kerman province, Iran. Methods: The anticholinesterase and antioxidant activity, as well as TPC and TFC of T. caramanicus hydroethanolic extracts, were evaluated using Ellman, DPPH, Folin-Ciocalteu, and aluminum chloride assays, respectively. Results: All extracts inhibited both acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzymes, with most showing a stronger effect on AChE. The extract T2-F exhibited the highest anti-AChE (IC50: 28.74 ± 0.21 µg/mL) and anti-BuChE (IC50: 65.49 ± 0.81 µg/mL) activities. The DPPH results indicated overall suitable antioxidant activity, with T4-F showing the highest antioxidant capacity (IC50: 16.22 ± 0.07 µg/mL), though about 4.2 times lower than ascorbic acid. The TPC (76.60 ± 0.98 - 211.73 ± 2.13 mg GAE/g extract) and TFC (19.15 ± 0.09 - 53.90 ± 0.13 mg QE/g extract) analyses revealed that T6-V and T5-F are rich sources of polyphenolic and flavonoid compounds. Pearson’s correlation analysis showed no direct correlation between TPC and TFC with their anticholinesterase and antioxidant activities. Conclusions: The hydroethanolic extracts of T. caramanicus demonstrated significant anticholinesterase and antioxidant activities, making them promising candidates for further studies aimed at developing therapeutic compounds with dual anticholinesterase and antioxidant properties, potentially for treating Alzheimer's disease (AD).

B-010 Intercomparison Study of Methods between Atellica Solution® and CI® Analyzers for Sodium, Potassium, and Chloride

Abstract Background Method verification is an essential analysis routinely conducted in accredited Clinical Laboratories whenever a change of method or instrument for a specific analyte is desired. Objectives The aim of the study is to verify that the results measured by the Atellica Solution® analyzer (Siemens) and the new CI® analyzer (Siemens) for Sodium, Potassium, and Chloride are interchangeable. Methods To conduct the intercomparison study of Sodium, Potassium, and Chloride, the techniques were properly calibrated and controlled on both analyzers. For each assay, 40 patient samples were selected, covering the entire measurement range, and analyzed by both instruments. To determine the degree of agreement between the instruments, a Bland Altman analysis of mean differences and a Passing Bablok regression were performed. Statistics are expressed with their 95% confidence intervals (CI). The evaluation of results was carried out using the statistical program Method Validator. Results In both the Bland-Altman and Passing Bablok regression analyses, a slight constant systematic error was detected for all three assays, as the 95% confidence interval of the observed mean difference and the 95% confidence interval of the intercept of the regression line do not include zero. No systematic proportional differences were observed based on the slope, as the 95% confidence interval includes the value one Conclusions The small constant systematic differences found in the analyses should be re-evaluated after performing a new calibration of the methods. In none of the cases are the differences clinically significant, as they do not exceed the total error established in our quality specification. Therefore, it can be concluded that the Atellica Solution® and CI 1900® analyzers are interchangeable for Sodium, Potassium, and Chloride assays

Investigating the antimicrobial and antibiofilm properties of marine halophilic Bacillus species against ESKAPE pathogens.

Antimicrobial resistance (AMR), known as the "silent pandemic," is exacerbated by pathogenic bacteria's ability to form biofilms. Marine compounds hold promise for novel antibacterial drug discovery. Two isolates from preliminary saltwater environment screening demonstrated antimicrobial activity and were subsequently identified as Bacillus subtilis MTUA2 and Bacillus velezensis MTUC2. Minimum inhibitory concentrations (MICs), minimum biofilm inhibition concentrations (MBICs) and minimum biofilm eradication concentrations (MBECs) required to prevent and/or disrupt bacterial growth and biofilm formation were established for MRSA, Staphylococcus aureus, Acinetobacter baumannii and Escherichia coli. The metabolic activity within biofilms was determined by the 2,3,5-triphenyltetrazolium chloride assay. Both Bacillus species exhibited unique antimicrobial effects, reducing MRSA and S. aureus planktonic cell growth by 50% and sessile cell growth for S. aureus and E. coli by 50% and 90%, respectively. No effect was observed against A. baumannii. Significant MBIC and MBEC values were achieved, with 99% inhibition and 90% reduction in MRSA and S. aureus biofilms. Additionally, 90% and 50% inhibition was observed in E. coli and A. baumannii biofilms, respectively, with a 50% reduction in E. coli biofilm. These findings suggest that the mode of action employed by B. subtilis MTUA2 and B. velezensis MTUC2 metabolites should be further characterized and could be beneficial if used independently or in combination with other treatments.

Mistletoe (Loranthus europaeus Jacq.): antioxidant, antimicrobial and anticholinesterase activities

Plants are natural resources responsible for different biological activities. Determining the biological activities of plants is very important. In our study, some biological activities and total phenolic and flavonoid contents of Loranthus europaeus Jacq. were determined. In this context, ethanol and methanol extracts of the plant were obtained using the soxhlet device. Total antioxidant, total oxidant, and oxidative stress statuses were determined using Rel Assay kits. Antimicrobial activity was tested against standard bacterial and fungal strains using the agar dilution test. Anticholinesterase activity was determined by detecting acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities. Determination of total phenolic content was determined using the Folin-Ciocalteu reagent. Total flavonoid quantification was performed using an aluminum chloride assay. As a result of the study, TAS values (Total antioxidant status) of ethanol and methanol extracts of the plant were determined as 5.620±0.134 and 6.384±0.134 mmol/L, respectively. TOS values (Total oxidant status) were determined as 10.997±0.183 and 13.368±0.222 µmol/L, respectively. OSI values (Oxidative stress index) were measured as 0.196±0.004 and 0.209±0.003, respectively. TPC (Total phenolic content) value was determined as 34.49±1.78 and 55.82±2.34 mg/g, respectively, and TFC (Total flavonoid content) value was determined as 46.10±2.40 and 55.49±1.15 mg/g, respectively. Ethanol and methanol extracts of the plant were found to be effective against bacterial and fungal strains at concentrations between 50-200 µg/mL. In addition, the anti-AChE values of ethanol and methanol extracts were determined as 13.51±0.81 and 22.79±1.86 µg/mL, respectively, and the anti-BChE values were determined as 27.84±0.62 and 33.08±1.63 µg/mL, respectively. According to the results obtained, it was determined that L. europaeus has antioxidant, antimicrobial, and anticholinesterase activity. In this context, it is thought that it can be used as a natural material in pharmacological designs.

Method for evaluation of Streptomyces growth and metabolism in the presence of glyphosate-based herbicide.

The use of pesticides, such as glyphosate, has increased due to population growth and the rising demand for food. Plant growth-promoting rhizobacteria (PGPR), such as Streptomyces, offer a more ecologically friendly alternative to the excessive use of pesticides. However, these bacteria undergo a complex life cycle involving the formation of hyphae, mycelia, and spores, which makes standardizing laboratory cultures challenging. In this context, we tested three methods for cultivating a Streptomyces isolate (CLV322) in the presence of the stressor agent glyphosate, denoted as M1, M2, and M3. These methods involved the simultaneous addition of the herbicide 24-48h after the start of cultivation. We evaluated the growth and cell viability of CLV322 using the 2,3,5-triphenyl tetrazolium chloride (TTC) assay under glyphosate-based herbicide stress (Roundup® Original DI) at concentrations ranging from 0.002 to 7.2mg mL- 1. We also assessed the ability of CLV322 to maintain PGPR characteristics in the presence of the herbicide by quantifying indolic compounds, siderophores, and phenazines. The cultivation method significantly influenced the production of metabolites by CLV322, with M3 yielding more consistent results across the evaluated parameters. Our findings suggest that germinating Streptomyces spores for 48h before introducing glyphosate (M3) enables the analysis of bacterial tolerance to herbicide stress. This methodology may also apply to evaluate other abiotic stresses on Streptomyces strains.

Determination of Quercetin in Iraqi Green Walnut Husk via Green Ionic Liquid-Based Digestion Extraction and Gel Filtration Procedures-Spectrophotometry and Microfluidic Paper-Based Analytical Platform Detection

في هذه الدراسة تم استخلاص ،عزل وتقدير الكيورستين الموجود في قشور الجوز العراقي باستخدام طريقة جديدة تعتمد على الاستخلاص بطريقة الهضم المبنية على استخدام السوائل الايونية و طريقة الترشيح بالجل للفصل المقترنة مع المطيافية الضوئية للتقدير ومن ثم مقارنتها مع تقنية جديدة تسمى تقنية السوائل المايكروية الورقية مستهدفة بناء طريقة اقتصادية، صديقة للبيئة وخضراء. ولقد تم اختيار 1 بيوتيل-3-مثيل ايمادزوليم كلورايد كمذيب اخضر بديل للمذيبات التقليدية بسبب خصائصة الذوبانية .ولقد اظهرت النتائج ان 100 ملغرام من القشورتحتاج الى 15 مل من المذيب و30 دقيقة من الهضم عند درجة75°C كظروف مثلى لاعطاء افضل ناتج للمستخلص بطريقة الهضم مقارنه مع طرق الاستخلاص الاخرى .تحت هذه الظروف المثلى تم عزل الكيورستين باستخدام طريقة الترشيح بالجل وتم التاكد من المركب المعزول باستخدام تقنية كروموتوغرافيا السائل عالية الاداء و اشعة فورية تحت الحمراء .تم تقدير الكيورستين المعزول من المستخلص لونيا ( طريقة كلوريد الالمنيوم) باستخدام طريقتي المطيافية الضوئية و وتقنية السوائل المايكروية الورقية معطيا الخطية لمنحنيات المعايرة لكلتا الطريقيتين بين 10-100 مليغرام / لتر مع معامل ارتباط0.9369 و 0.7917 لكلتا الطريقتين. ومن ثم تم تقدير الفعالية المضادة للاكسدة باستخجام طريقة قوة الاختزال للمركب المعزول مظهرة فعالية مضادة للاكسدة جيدة و بين تحليل التباين (ANOVA) عدم وجود فرق كبير بين تقنية المطيافية الضوئية وتقنية السوائل المايكروية الورقية. كخلاصة يمكن اعتبار المذيب الايوني BMIMCLمذيبا امنا ومناسبا لاستخلاص المركبات النباتية ذات الخواص الطبية وكذلك فان تقنية السوائل المايكروية الورقية اثبتت مقدرتها على التحليل بكفاءة مساوية الى طرق التحليل التقليدية بالاضافة الى خصائصها الخضراء.

A case report of elevated bromide levels from pyridostigmine bromide for treatment of myasthenia gravis.

In an effort to expedite the publication of articles, AJHP is posting manuscripts online as soon as possible after acceptance. Accepted manuscripts have been peer-reviewed and copyedited, but are posted online before technical formatting and author proofing. These manuscripts are not the final version of record and will be replaced with the final article (formatted per AJHP style and proofed by the authors) at a later time. Elevated serum bromide levels can cause dermatological, gastrointestinal, and neurological abnormalities. As bromide and chloride are both halogens, bromide may interfere with chloride assays, causing a falsely high serum chloride concentration and a low or negative anion gap. There is a paucity of data describing bromide toxicity from high doses of pyridostigmine bromide (PB). This case report describes a patient with an elevated bromide level with neurological symptoms from a therapeutic dose of PB. A 37-year-old male with myasthenia gravis secondary to type B2 thymoma status following thymectomy presented in myasthenic crisis. He required mechanical ventilation and was managed with steroids, intravenous immune globulin, plasmapheresis, and PB. On day 9, the patient experienced acute agitation. He had an anion gap of 2 mEq/L and a chloride concentration of 109 mEq/L. The plasma creatinine concentration was 0.63 to 1.15mg/dL and urine output was 0.76 to 1.79mL/kg/h throughout his admission. All other laboratory values were normal. The daily dose of PB was 660mg on day 9, but the patient received 76mg of intravenous PB over the first few days of his admission with the largest dose in 24 hours equal to 48mg. On day 10, the patient's bromide level was 37 μg/mL. His agitation was initially managed with quetiapine, followed by PB dose reduction. To our knowledge, there are 2 cases in the literature of bromide toxicity secondary to PB. These patients experienced neurological symptoms with bromide levels of 88 to 90 μg/mL. Bromide concentrations of more than 12 μg/mL are associated with a higher risk of neuronal dysfunction demonstrated as disturbances on an electroencephalogram, and levels greater than 50 μg/mL are considered toxic. While our patient's bromide level was not as high as those previously reported, no other causes for his agitation were identified. Elevated bromide levels from therapeutic PB can occur, and monitoring of these levels should be considered.

Phytochemical, proximate analysis and antioxidant activity of the rhizome of Alpinia nigra (Gaertn.) B.L. Burtt (Zingiberaceae) in Tamulpur district, Assam

The present research examines the chemical composition, proximate characteristics and antioxidant analysis of Alpinia nigra’s rhizome extract. Results of GC-MS analysis revealed a total of 22 volatile compounds of which Tris (tert-butyldimethylsilyloxy) arsane (2.89 %), 2-ethylthiolane, S, S-dioxide (3.47 %), Ambrial (6.68 %) and Silicic acid, diethyl bis (trimethylsilyl) ester (4.59 %) were the major compounds. The rhizome was found to have variable quantities of proximate compositions such as moisture content (55.5 %), ash content (4.72 %), crude fat (52 %), crude fibre (41.25 %), crude protein (28.95 µg/g), carbohydrate (46.27 %) and dry matter (33.4 %). The crude extraction of the rhizome was done by Soxhlet apparatus. The phytochemical screening showed the presence of alkaloid, tannin, terpenoid, glycoside, steroid, carbohydrate and a considerable amount of phenol and flavonoid. Phenolic and flavonoid content in the extracts was determined by the Folin-Ciocalteu and aluminium chloride assays as gallic acid equivalent and quercetin equivalent respectively. To evaluate the antioxidant activity of rhizome extract, phosphomolybdenum assay, ferric-reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picryl-hydrazil (DPPH) were used. The findings indicate that the rhizome extract is rich in phenol and flavonoid content and would serve as a promising source of natural antioxidants in the food, cosmetics and pharmaceutical industries.

Antioxidant, Antimicrobial, and Antialzheimer Activities of Tagetes patula (Asteraceae)

Plants are natural products used in the prevention and treatment of many diseases. In this study, antioxidant, antimicrobial, anticholinesterase activities and total phenolic and flavonoid contents of Tagetes patula L. samples collected from Iraq were determined. The aerial parts of the plant were extracted with ethanol in a soxhlet device. The antioxidant potential of the extracts was measured with Rel Assay kits. Antimicrobial activity was determined by the agar dilution method against standard bacterial and fungal strains. To determine anticholinesterase activity acetyl and butyrylcholinesterase inhibitions were tested. The total phenolic content of the samples was carried out using the Folin-Ciocalteu reagent. The quantification of flavonoids was conducted using an aluminum chloride assay. As a result of the analyses, the total antioxidant value of the plant extract was determined as 5.386±0.142 mmol Trolox equiv./L, the total oxidant value was 8.287±0.146 μmol H2O2 equiv./L and the oxidative stress index was determined as 0.154±0.003. Plant extracts showed the highest activity against Candida species. It was also effective against bacterial and fungal strains at concentrations between 50-400 µg/mL. Acetylcholinesterase activity of the plant extract was determined as 24.97±0.98, and butyrylcholinesterase activity was determined as 35.65±0.94. Additionally, its total phenolic content was determined as 63.64±0.74 mgGAE/g and its total flavonoid content was 108.9±1.55 mgQE/g. It has been determined that the plant has antioxidant, antimicrobial, and antiallergic potential.

Photodynamic antibacterial research on hypericin-loaded PEGylated mesoporous silica delivery system

In this study, a novel drug delivery system (MSN-PEG-Hypericin) was successfully fabricated using tetraethyl orthosilicate and 3-aminopropyltriethoxysilane as raw materials, and the PEGylation of the prepared aminated mesoporous silica and grafting of hypericin onto the carrier were further conducted to obtain MSN-PEG-Hypericin. The successful preparation of MSN-PEG-Hypericin was characterized by several physical-chemical techniques. Furthermore, the MSN-PEG-Hypericin system increased the ability of hypericin to generate reactive oxygen species (ROS) in vitro. The cytotoxicity assay and hemolysis analysis showed that MSN-PEG-Hypericin had good biocompatibility. For antibacterial studies, the irradiation time and incubation time of photodynamic therapy (PDT) for S. aureus and E. coli were respectively 8 min and 8 h, and the concentrations of hypericin were 2.5 and 5 μg/mL. The result of triphenyl tetrazolium chloride assay indicated that MSN-PEG-Hypericin had stronger photodynamic antibacterial activity than free hypericin, and S. aureus was more sensitive to PDT than E. coli, which was related to their cell structural differences. The antibacterial mechanism study indicated that the generated ROS could destroy the bacterial structures and cause bacterial death due to the leakage of the contents. The MSN-PEG-Hypericin system prepared in this study had potential application prospects in the antibacterial field.

Ischemic stroke outcome after promoting CD4+CD25+ Treg cell migration through CCR4 overexpression in a tMCAO animal model

The importance of neuroinflammation during the ischemic stroke has been extensively studied. The role of CD4+CD25+ regulatory T (Treg) cells during the recovery phase have shown infarct size reduction and functional improvement, possibly through the mitigation of inflammatory immune responses. We aimed to investigate the molecular factors involved in microglia-Treg cell communication that result in Treg trafficking. First, we observed the migration patterns of CD8+ (cytotoxic) T cells and Treg cells and then searched for chemokines released by activated microglia in an oxygen–glucose deprivation (OGD) model. The transwell migration assay showed increased migration into OGD media for both cell types, in agreement with the increase in chemokines involved in immune cell trafficking from the mouse chemokine profiling array. MSCV retrovirus was transduced to overexpress CCR4 in Treg cells. CCR4-overexpressed Treg cells were injected into the mouse transient middle cerebral artery occlusion (tMCAO) model to evaluate the therapeutic potential via the tetrazolium chloride (TTC) assay and behavioral tests. A general improvement in the prognosis of animals after tMCAO was observed. Our results suggest the increased mobility of CCR4-overexpressed Treg cells in response to microglia-derived chemokines in vitro and the therapeutic potential of Treg cells with increased mobility in cellular therapy.

Theophylline as a quorum sensing and biofilm inhibitor in Pseudomonas aeruginosa and Chromobacterium violaceum.

Quorum sensing (QS) is pivotal in coordinating virulence factors and biofilm formation in various pathogenic bacteria, making it a prime target for disrupting bacterial communication. Pseudomonas aeruginosais a member of the "ESKAPE" group of bacterial pathogens known for their association with antimicrobial resistance and biofilm formation. The current antibiotic arsenal falls short of addressing biofilm-related infections effectively, highlighting the urgent need for novel therapeutic agents. In this study, we explored the anti-QS and anti-biofilm properties of theophylline against two significant pathogens, Chromobacterium violaceum and P. aeruginosa. The production of violacein, pyocyanin, rhamnolipid, and protease was carried out, along with the evaluation of biofilm formation through methods including crystal violet staining, triphenyl tetrazolium chloride assay, and fluorescence microscopy. Furthermore, computational analyses were conducted to predict the targets of theophylline in the QS pathways of P. aeruginosa andC.violaceum. Our study demonstrated that theophylline effectively inhibits QS activity and biofilm formation inC. violaceumandP. aeruginosa. InP. aeruginosa, theophylline inhibited the production of key virulence factors, including pyocyanin, rhamnolipid, protease, and biofilm formation. The computational analyses suggest that theophylline exhibits robust binding affinity to CviR inC. violaceumand RhlR inP. aeruginosa, key participants in the QS-mediated biofilm pathways. Furthermore, theophylline also displays promising interactions with LasR and QscR inP. aeruginosa. Our study highlights theophylline as a versatile anti-QS agent and offers a promising avenue for future research to develop novel therapeutic strategies against biofilm-associated infections.

Gynostemma pentaphyllum Makino exerts cardioprotective effects by improving hemodynamic, biochemical and histopathological changes through activation of PI3K signalling in isoproterenol-induced myocardial infarction in rats.

To evaluate the cardioprotective effects of Gynostemma pentaphyllum Makino in isoproterenol-induced myocardial infarction in rats and to evaluate the role of phosphatidylinositol 3-kinases (PI3K) in cardioprotection. The protective effect of the hydroalcoholic leaf extract of Gynostemma pentaphyllum (LEGP) on the heart was investigated against isoproterenol (ISO)-induced MI in rats. Preliminary phytochemical screening was performed followed by molecular docking. For the in vivo studies Wistar albino rats (Male) were divided among different groups. Different parameters were evaluated such as heart weight index, Electrocardiogram (ECG) analysis, triphenyl tetrazolium chloride assay, cardiac enzyme markers, oxidative stress, antioxidant enzymes, PI3K levels, and histopathology of cardiac tissue. Results showed that LEGP improved the electrocardiogram, reduced infarct size, and decreased the levels of cardiac enzyme markers and oxidative stress, while antioxidant enzymes and PI3K levels were increased. LEGP protected the heart against ISO-induced MI in rats by improving hemodynamic, biochemical and histological attributes. These protective effects were produced by the phytoconstituents of the LEGP through modulation of the PI3K signalling pathway.

Amalgamation, characterization and biological assay of benzyltrimethylammonium chloride

The year 2020 appeared to be a massive loss of human population due to the COVID-19 virus. During that period of time, different medicines were utilised as disinfectants to minimise the spread of the virus. Among those, quaternary ammonium compounds were the most likely to be utilised as disinfectants. In this study, a quaternary ammonium compound namely benzyl-trimethylammonium chloride was synthesised at 0.1N to 0.8N concentrations and tested to find the activities of bacteria and fungi. Since this compound prosperously acts against bacteria and fungi, it can also truncate the spread of diseases caused by viruses. Research has proved that if an anti-microbial solution has the faculty to kill bacteria, then it also has viral activity. Then this solution is dissolved with protein powder and studied against protein solubility test. Furthermore, in this research, FTIR and UV spectral analyses of benzyl-trimethylammonium chloride were taken and explicated in brief regarding its functional group and absorption wavelength. This quaternary ammonium compound is very useful for mankind.

Antioxidant, antimicrobial and anticholinesterase activity of Dittrichia graveolens

Plants are responsible for many different biological activities. In our study, antioxidant, antimicrobial and anticholinestrase activities of Dittrichia graveolens (L.) Greuter were determined. In addition, the total phenolic and flavonoid contents of the plant were measured. The aerial parts of the plant were extracted with ethanol. Antioxidant activities of the extracts were measured with Rel assay kits. Antimicrobial activity was determined using the agar dilution method. Anticholinesterase activity was determined by measuring acetyl and butyrylcholinesterase activities. Determination of total phenolic content was determined using the Folin-Ciocalteu reagent. Total flavonoid quantification was performed using aluminum chloride assay. As a result of the study, the TAS value of the plant was determined as 6.933±0.121, the TOS value was 12.535±0.244 and the OSI value was 0.181±0.006. It was determined that the plant extract was effective against microorganisms at concentrations of 50-400 µg/mL. Additionally, the anti-AChE value was found to be 25.88±1.73 and the anti-BChE value was 45.32±2.26. Total phenolic content was determined as 86.42±2.72, and total flavonoid content was determined as 117.96±1.93. As a result, it was determined that D. graveolens has antioxidant, antimicrobial and anticholinesterase activity.