Blue Method Research Articles

The effect of cardiac rehabilitation prescription on medication complications and ET-1, WMSI in elderly patients with coronary heart disease.

To explore the effects of cardiac rehabilitation prescriptions on medication complications, Endothelin 1 (ET-1), and Wall Motion Score Index (WMSI) in elderly patients with coronary heart disease. 98 elderly patients with coronary heart disease admitted to the Department of Cardiology of a hospital from January 2020 to June 2022 are selected. According to the red and blue ball method, 98 research subjects are divided into a control group and an observation group. The control group receives routine treatment and exercise intervention, while the observation group receives cardiac rehabilitation prescriptions. After a follow-up of 6 months, the incidence of medication complications between the two groups is compared. The changes in ET-1 and WMSI levels are compared before treatment, 1 month, 3 months, and 6 months after treatment. The two groups had no significant difference in ET-1 and WMSI levels before intervention. After intervention, both groups showed significant reductions in ET-1 and WMSI compared with baseline. The ET-1 of the observation group decreased significantly faster than the control group after 1 month, 3 months, and 6 months of intervention. The WMSI decreased more significantly at 6 months. The repeated measurement variance showed that there was a significant difference in the trend of ET-1 changes between the two groups, while the difference between the WMSI groups was not significant. The incidence of medication complications in the observation group was 10.20%, significantly lower than the 26.53% in the control group. Cardiac rehabilitation prescriptions can reduce the incidence of medication complications in elderly patients with coronary heart disease. It can reduce ET-1 levels faster, improve myocardial motor function, which has better myocardial function and medication safety than conventional treatment.

Refined Surface Area Determination of Graphene Oxide Using Methylene Blue as a Probe Molecule: A Comparative Approach

Abstract In this research, we explored the effectiveness of the methylene blue (MB) adsorption method as an alternative approach for determining the specific surface area of graphene oxide (GO). Initially, through a comparative analysis with reference activated carbon (AC), we identified the limitations of utilizing N2 physisorption for specific surface area determination of GO. Our findings revealed that the standard pretreatment process (heating under vacuum) before N2 physisorption led to damage to the surface oxygen groups on GO, and the measured surface areas (43 m2/g) do not accurately represent the entire surface area. To optimize MB coverage on GO, we conducted adsorption equilibrium experiments, focusing on controlling temperature and pH. The pH was significantly important in regulating the coverage of MB. Under the optimized MB adsorption conditions, the specific surface area of GO was 1555 m2/g. Our assumptions regarding specific surface area calculations were supported by structural characterization of samples with varying MB uptakes. The results confirmed a uniform coverage of MB on GO by SEM-EDX, XRD, and AFM.

Evaluation of Selected Plant Phenolics via Beta-Secretase-1 Inhibition, Molecular Docking, and Gene Expression Related to Alzheimer’s Disease

Background: The goal of the current study was to investigate the inhibitory activity of six phenolic compounds, i.e., rosmarinic acid, gallic acid, oleuropein, epigallocatechin gallate (EGCG), 3-hydroxytyrosol, and quercetin, against β-site amyloid precursor protein cleaving enzyme-1 (BACE1), also known as β-secretase or memapsin 2, which is implicated in the pathogenesis of Alzheimer’s disease (AD). Methods and Results: The inhibitory potential against BACE1, molecular docking simulations, as well as neurotoxicity and the effect on the AD-related gene expression of the selected phenolics were tested. BACE1 inhibitory activity was carried out using the ELISA microplate assay via fluorescence resonance energy transfer (FRET) technology. Molecular docking experiments were performed in the human BACE1 active site (PDB code: 2WJO). Neurotoxicity of the compounds was carried out in SH-SY5Y, a human neuroblastoma cell line, by the Alamar Blue method. A gene expression analysis of the compounds on fourteen genes linked to AD was conducted using the real-time polymerase chain reaction (RT-PCR) method. Rosmarinic acid, EGCG, oleuropein, and quercetin (also used as the reference) were able to inhibit BACE1 with their respective IC50 values 4.06 ± 0.68, 1.62 ± 0.12, 9.87 ± 1.01, and 3.16 ± 0.30 mM. The inhibitory compounds were observed to occupy the non-catalytic site of the BACE1. However, hydrogen bonds were found to be present between rosmarinic acid and EGCG and aspartic amino acid D228 in the catalytic site. Oleuropein and quercetin effectively suppressed the expression of PSEN, APOE, and CLU, which are recognized to be linked to the pathogenesis of AD. Conclusions: The outcomes of the work bring quercetin, EGCG, and rosmarinic acid to the forefront as promising BACE1 inhibitors.

Exploring methylene blue as an effective alternative to radiotracer for sentinel lymph node biopsy in early breast cancer in resource-limited settings

Background: Sentinel lymph node biopsy is the recommended approach in the evaluation of axilla during breast cancer surgery. This study evaluated the results of patients who underwent methylene blue sentinel lymph node biopsy. Methods: The study included 35 female patients with T1, T2 and T3 tumours. 5 ml of 1% methylene blue was injected into the periareolar region. The axillary sentinel lymph node was found and removed, and axillary dissection was performed. The sentinel lymph node and axillary dissection specimen were histopathologically examined, and the results were compared. Results: The sentinel lymph node was found in 33 (90%) patients. There were no complications associated with the use of methylene blue dye. Methylene blue-stained sentinel nodes were histologically negative for malignancy in 42% and positive in 58% of patients. Corresponding to sentinel nodes, the final axillary staging was negative in 60% and positive in 40% of patients. On diagnostic accuracy parameters, the sensitivity of the methylene blue method in early breast cancer (cT1-3N0M0) patients was 90.47%, and the specificity was 100%. The PPV of the technique under study was found to be 100% and NPV was 85.71%. The accuracy rate was 93%, and the false negative rate was identified as 9.5%. Conclusions: Sentinel lymph node biopsy by methylene blue is a method that can be applied with high accuracy. Methylene blue can be considered as an alternative to radio colloid dye in sentinel lymph node biopsy in a limited resource setting.

Does gut microbiota dysbiosis impact the metabolic alterations of hydrogen sulfide and lanthionine in patients with chronic kidney disease?

BackgroundChronic Kidney Disease (CKD) is characterized by a methionine-related metabolic disorder involving reduced plasma levels of hydrogen sulfide (H2S) and increased lanthionine. The gut microbiota influences methionine metabolism, potentially impacting sulfur metabolite dysfunctions in CKD. We evaluated whether gut microbiota dysbiosis contributes to H2S and lanthionine metabolic alterations in CKD.MethodsThe gut microbiota of 88 CKD patients (non-dialysis, hemodialysis, and transplant patients) and 26 healthy controls were profiled using 16 S-amplicon sequencing. H2S and lanthionine concentrations were measured in serum and fecal samples using the methylene blue method and LC-MS/MS, respectively.ResultsThe CKD population exhibited a tenfold increase in serum lanthionine associated with kidney dysfunction. Despite lanthionine retention, hemodialysis and transplant patients had significantly lower serum H2S than healthy controls. Fecal H2S levels were not altered or related to bloodstream H2S concentrations. Conversely, fecal lanthionine was significantly increased in CKD compared to healthy controls and associated with kidney dysfunction. Microbiota composition varied among CKD groups and healthy controls, with the greatest dissimilarity observed between hemodialysis and transplant patients. Changes relative to the healthy group included uneven Ruminococcus gnavus distribution (higher in transplant patients and lower in non-dialysis CKD patients), reduced abundance of the short-chain fatty acid-producing bacteria Alistipes indistinctus and Coprococcus eutactus among transplant patients, and depleted Streptococcus salivarius in non-dialysis CKD patients. A higher abundance of Methanobrevibacter smithii, Christensenella minuta, and Negativibacillus massiliensis differentiated hemodialysis patients from controls. No correlation was found between differentially abundant species and the metabolic profile that could account for the H2S and lanthionine alterations observed.ConclusionsThe metabolic deregulation of H2S and lanthionine observed in the study was not associated with alterations in the gut microbiota composition in CKD patients. Further research on microbial sulfur pathways may provide a better understanding of the role of gut microbiota in maintaining H2S and lanthionine homeostasis.

Electroacupuncture preconditioning prevents urticaria by regulating inflammatory response in rats with urticaria

To observe the effect of electroacupuncture (EA) preconditioning at "Quchi" (LI11) and "Xuehai" (SP10) in prevention of urticaria. Twenty-four male SD rats were randomly divided into control, model and preconditioning of EA (Pre-EA) groups (8 rats/group). The urticaria model was established by intradermal injection of dilute allogeneic antioalbumin serum at the spots of the bilateral symmetry of the spine on the back, and followed by tail venous injection of mixture solution of egg albumin diluent, plus 0.5% Evans blue and normal saline. Ten days before the end of modeling, rats of the pre-EA group received EA stimulation of LI11 and SP10 for 20 min, once a day for 10 consecutive days. The times of rat's scratching the sensitized skin were recorded. HE staining method was used to observe the pathological changes of skin tissue, and toluidine blue staining method was used to observe the morphology of mast cells (MCs) in the skin, blood, mesentery, and peritoneal fluid, and calculate the degranulation rate. Immunohistochemical stainning was used to detect immunoglobulin E (IgE), histamine (HIS), and 5-hydroxytryptamine (5-HT) expressions in subcutaneous tissue. NOD like receptor thermal domain associated protein 3 (NLRP3) inflammasome, apoptosis related granule protein (ASC), and cysteine aspartate aminotransferase 1 (Caspase-1) protein expression levels in skin tissue were detected by Western blot. The contents of serum interleukin(IL)-1β and IL-18 were detected using ELISA method. Compared with the control group, the scratching times, amount of Evans blue exudation of the sensitized blue spots, degranulation rate of MCs in skin, blood, mesentery and peritoneal fluid, the expression levels of IgE, HIS, 5-HT in subcutaneous tissue, protein expression levels of NLRP3, ASC, Caspase-1 in skin tissue, and the contents of serum IL-1β and IL-18 were significantly increased (P<0.01, P<0.05) in the model group. In comparison with the model group, the scratching times, amount of Evans blue exudation of the sensitized blue spots, degranulation rate of MCs, the expression levels of IgE, HIS, 5-HT in subcutaneous tissue, protein expression levels of NLRP3, ASC, Caspase-1 in skin tissue, and the contents of serum IL-1β and IL-18 in EA group were significantly decreased (P<0.01, P<0.05). EA preconditioning at LI11 and SP10 can prevent and treat UR by inhibiting inflammatory response, which is related to the regulation of pyroptosis.

Antioxidant and Anti-Proliferative Effects of Ornithogalum balansae Extracts Collected in Black Sea Region Against Lung Cancer Cells

Ornithogalum is a genus of wild herb species widely used as food and in traditional medicine. This study investigated some of the biologically active properties of Ornithogalum balansae grown in the eastern Black Sea region of Türkiye. The investigations were carried out using the methanolic extracts of the plant’s aerial and bulbs (B1 and B2) parts. The phenolic composition was examined as total phenolic content (TPC), total flavonoid content (TFC), and HPLC-PDA. The antioxidant capacity of the extracts was tested based on ferric reducing/antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging methods. The anti-proliferative activity was tested against metastatic cell lines H1299 and H209, non-small-cell lung A549, and fibroblast MRC-5 cell lines using MTT and trypan blue methods. Wound-healing and invasion chamber assays were used to determine the inhibitory effects of the extracts on migration and invasion, respectively. The extract of the aerial part contained a large number of phenolic substances and high antioxidant capacity. The extract exhibited a significant anti-proliferative effect on the human lung cancer cells (A549 and H209), with IC50 values of 0.97 ± 0.04 and 1.06 ± 0.07 µg/mL, respectively. Moreover, the aerial part exhibited inhibition of migratory and invasive capacities in A549 cells at a concentration of 1.50 µg/mL. The findings associated with O. balansae suggest a promising therapeutic potential against lung cancer cells.

Yigong San improves learning and memory functions of APP/PS1 transgenic mice by regulating brain fluid metabolism

To explore the mechanism by which Yigong San (YGS) improves learning and memory abilities of APP/PS1 transgenic mice in light of cerebral fluid metabolism regulation. Three-month-old male APP/PS1 transgenic mice and wild-type C57BL/6 mice were both randomized into control group, model group, donepezil (1.67 mg/kg) group, and YGS (7.5 g/kg) group and received the corresponding treatments via gavage once daily for one month. After the treatments, the mice were assessed for learning and memory functions using Morris water maze test and examined for hippocampal and cortical pathologies and amyloid plaques using HE, immunohistochemical and thioflavin S staining; ELISA and Evans blue method were used for detecting Aβ1-40 and Aβ1-42 levels in the brain tissue and serum and assessing blood-brain barrier (BBB) integrity. Immunofluorescence colocalization was used to investigate AQP4 polarization on astrocytes. Western blotting was performed to detect the expressions of VE-cadherin, ZO-1, occludin, β-amyloid precursor protein (APP), BACE1, insulin-degrading enzyme (IDE), LRP1, RAGE, and AQP4 proteins. Compared with the control mice, APP/PS1 mice showed significant impairment of learning and memory abilities, increased degeneration or necrosis of hippocampal and cortical neurons, pathological scores, Aβ-positive plaques, elevated Aβ1-40 and Aβ1-42 levels in the brain tissue and serum, increased BBB permeability, upregulated RAGE expression, lowered expressions of VE-cadherin, LRP1, ZO-1, occludin, and AQP4 proteins, and reduced AQP4- expressing GFAP-positive cells. YGS treatment significantly improved the performance of the transgenic mice in Morris water maze test, reduced hippocampal and cortical pathologies and Aβ-positive plaques, and ameliorated the abnormal changes in Aβ1-40 and Aβ1-42 levels, BBB permeability, protein expressions of RAGE, VE-cadherin, LRP1, ZO-1, occludin and AQP4, and the number of AQP4-expressing GFAP-positive cells. YGS improves learning and memory changes in APP/PS1 mice by ameliorating neuronal damage and Aβ pathology in the brain and regulating brain fluid metabolism.

Comprehensive Analysis of Needle Bearing Failures in Fuel Transfer Pumps: Insights from Testing and Finite Element Analysis

&lt;div&gt;This study investigates the failure mechanisms of needle bearings within fuel transfer pump assemblies through a comprehensive approach combining endurance testing, detailed inspection, the Dykem blue method, proximity sensors, and finite element analysis (FEA). The findings reveal critical insights into the causes of failure, highlighting significant axial displacement, with a maximum of 0.37 mm measured by proximity sensors. The Dykem technique identified distinct wear patterns across various components, pinpointing areas of high stress and potential failure. Detailed bearing inspections uncovered trunnion damage and abrasive wear, corroborated by FEA, which quantified displacements of 0.144 mm in the x-direction, 0.030 mm in the y-direction, and 0.015 mm in the z-direction. The primary operational factors contributing to bearing failure were contamination and inadequate axial control. These insights are pivotal, as they align with and expand upon established literature on bearing failures, providing a deeper understanding of the interplay between mechanical wear and operational conditions. Despite the robustness of the methodology, challenges included ensuring the accuracy of axial displacement measurements and replicating real-world operational stresses in a controlled environment. The study proposes several recommendations to enhance axial support and optimize system design to mitigate the identified issues. The societal impact of this research is significant, offering potential improvements in machinery reliability, which can lead to enhanced industrial efficiency and safety standards. This work advances the current knowledge in the field and provides practical solutions for extending the lifespan and performance of critical mechanical components in fuel transfer systems.&lt;/div&gt;

Determination of Cortisol Hormone from Sweat Samples and Interpretation with Microcontroller

Cortisol, known as the body's stress hormone, regulates metabolism, inflammation, and the immune system. It affects various bodily systems such as the nervous, immune, cardiovascular, respiratory, reproductive, and musculoskeletal systems. Normal sweat cortisol levels range from 8 to 142 ng/ml, with elevated levels indicating stress (200 ng/ml in women, 500 ng/ml in men). This study focuses on colorimetric cortisol analysis using a microfluidic layer to collect sweat samples. Samples are mixed with a specific color reagent. Cortisol levels are quantitatively analyzed using Z-Score values and RGB channel density via photographs taken with the ESP 32 module. The aim is to determine cortisol levels, aiding in diagnosing conditions like Cushing's syndrome and Addison's Disease, leveraging wearable technology. The developed system integrates a microcontroller into the wearable microfluidic system for cortisol measurements throughout the day. Data is displayed and recorded on a computer for streamlined cortisol monitoring. The blue tetrazolium method used offers stable colors and rapid reactions (10 minutes). The Limit of Detection (LOD) is 0.3 ug/mL, with a detection limit of 1.1 ug/mL. Combined with the ESP 32 Cam module, the system detects cortisol concentrations from 0.8 – 60 ug/mL.

Enhancing the effectiveness of teaching neuroanatomy: A comparative study using stained and unstained brain sections to interpret cross sectional neuroanatomy

BackgroundNeuroanatomy is considered one of the most challenging parts of anatomy curriculum and the complexity in understanding neuroanatomy generates from the lack of a 3-dimensional view of the structures. Since everything appears grey and white and is difficult to differentiate the various structures in a cross section of brain, staining aids in the visual interpretation and retention. There are various staining methods employed such as Mulligan's method, Alston's method, Prussian blue reaction method, however the results obtained by Alston's method was the best as gathered from previous literature. Material and methodsThe study was conducted in the department of Anatomy, formalin fixed brain slices measuring 10 mm thickness were stained using Alston's method. 250 medical students of the first professional year were demonstrated features such as the internal capsule, caudate nucleus, lentiform and thalamus in the stained and unstained transverse sections of cerebrum during their neuroanatomy practical sessions. Following which a feedback was collected regarding both the specimens using Microsoft Forms. ResultsThere was a positive feedback from the students with 83 % of them preferring the stained over the unstained section. Excerpts from the open ended question where all in favour of the stained sections -“much easier to see and identify the parts in the stained section because of the clear distinction between white and grey matter”. ConclusionsMacroscopic staining of sections of the brain using Alston's method could be implemented as a valuable method for effective teaching of neuroanatomy since students appreciated the structures better in the stained sections which enhanced their retention of neuroanatomy. Since Alston stain produces the least shrinkage, these stained sections could be utilized for research studies and such specimens could also be plastinated.

Dellaglioa Algida Cell-Free Supernatant Inhibits Pseudomonas Fluorescence and Pseudomonas Fragi by Destroying Cell Membranes.

The aim of this study was to examine the components of the cell-free supernatant (CFS) derived from a novel strain of psychrophilic Lactobacillus, Dellaglioa algida, and to further elucidate the impact of this CFS on various cellular processes. Specifically, we sought to understand its effects on the cell membrane, protein and DNA release, protease activity, and metabolites of Pseudomonas fluorescens and Pseudomonas fragi, thereby clarifying the antibacterial mechanism involved. The CFS components were analyzed using Gas Chromatography-Mass Spectrometry (GC-MS), the Coomassie Brilliant Blue method, and the phenol-sulfuric acid method. The inhibitory effect of the CFS on Pseudomonas fluorescens and Pseudomonas fragi was assessed using the ethidium bromide (EB) assay, Oxford cup assay, and ultramicroassay. Additionally, we analyzed the metabolites produced by Pseudomonas fluorescens and Pseudomonas fragi when treated with the CFS. The findings reveal that the CFS of Dellaglioa algida contains 94 volatile components, with protein and sugar concentrations of 32.857 ± 0.9705 mg/mL and 98.250 ± 4.210 mg/L, respectively. The CFS induces varying degrees of damage to the cell membranes of both Pseudomonas fluorescens and Pseudomonas fragi, leading to the release of intracellular proteins and DNA. Furthermore, the CFS reduced the protease activity and metabolic capacity of Pseudomonas fluorescens and Pseudomonas fragi. These results enhance our understanding of the mechanism by which psychrophilic Dellaglioa algida inhibits Pseudomonas fluorescens and Pseudomonas fragi, confirming that its inhibitory effect predominantly occurs through damage to the biological cell membranes of Pseudomonas. Dellaglioa algida is a newly identified cold-adapted inhibitor of Pseudomonas, indicating that its CFS is an effective microbial inhibitor in cold environments. This discovery suggests potential applications in inhibiting the growth and reproduction of Pseudomonas fluorescens and Pseudomonas fragi in food, pharmaceuticals, perfumes, and other chemicals, providing a valuable new reference for industrial preservation.

Evolving nature-based solutions for Australia’s Indigenous estate in 2024 – opportunities and challenges

Globally, there is growing interest and potential for investment in Nature-based Solutions (NbS) to protect, manage or restore ecosystems through incentive schemes including Payment for Ecosystem Services (PES), Nature Repair, Carbon Markets, Common Asset Trusts. Collectively, these NbS markets establish interchangeable mechanisms to help address biodiversity and climate crises, as well as socio-economic issues concerning many Indigenous Peoples and Local Communities (IPLCs). IPLCs manage and/or own almost 32% of the world’s land area through customary and community-based tenure arrangements. Since 2000 several PES schemes have been implemented in Australia. These schemes have been overwhelmingly implemented as public-good expenditure, with governments providing ~90% of total funding. Indigenous people have either the legal right to run or veto a carbon market or nature repair project over 57% of the Australian land mass, increasing to 63% in savanna and 85% in desert regions. Here we critically assess opportunities and challenges for Australia’s Indigenous estate associated with existing and emerging NbS market approaches, especially the Commonwealth’s current Savanna Fire Management (SFM) and Human Induced Regeneration methods, evolving Integrated Farm &amp; Land Management (IFLM) and Blue Carbon methods, and proposed Nature Repair (NR) market. To date, Indigenous NbS interests have focused especially on SFM across northern Australia, with prospective opportunities especially for IFLM and NR markets. Most available schemes focus on remediation of degraded lands and seas, ignoring cost-effective investment opportunities to maintain habitats and ecosystems in less-degraded condition. Government-supported Common Asset Trusts can provide effective models for governance of stewardship schemes relevant to on-going care for relatively intact ecosystems. In Discussion we summarise key methodological, institutional, and policy opportunities and challenges for constructive Indigenous engagement with developing NbS markets. Our purpose is to provide an Indigenous land and sea management context to inform development of rapidly evolving NbS markets in Australia.

Phosphorous intake in foods and phosphorus status markers in circulation in the Boston Puerto Rican Health Study

Phosphorus (P) additives may be deleterious for health. We measured the P content of key foods, and associations of P intake with biomarkers in the Boston Puerto Rican Health Study (BPRHS). Direct chemical analysis of 92 foods was done with the molybdenum blue spectrophotometric method and inductively coupled plasma mass spectrometry (ICP-MS). A novel algorithm was used to determine bioavailable, natural, and added P. We estimated P intakes from foods in 1323 participants, aged 45–75 y, and associations of these with serum P, fibroblast growth factor 23 (FGF23), parathyroid hormone (PTH), and Klotho. Relationships between intakes and status markers were assessed with Pearson’s correlations and t-tests. Our food analyses generally support P values in the USDA nutrient database, with the exceptions of American and cheddar cheese, which had more P than in the database. Women had higher added P intake than men, and younger participants had higher added P than those older. Total P intake tended to be positively associated with serum P and klotho, and inversely associated with PTH, but relationships were not strong. Puerto Rican adults have high intake of additive P. Culturally sensitive interventions that highlight dietary quality are needed.

Ultrasonic assisted adsorption of methylene blue using blood clam shell as a low-cost adsorbent

Methylene blue is a synthetic dye used in various industries, such as wood, linen, and silk. Generally, as much as 10–200 mg/L of methylene blue is released into the environment and potentially polluting the environment. One of methylene blue removal method from wastewater is adsorption, particularly using blood clam (Anadara granosa) shells adsorbent. Blood clam shells contain CaCO3 (up to 98.7 %) that can be utilized to adsorb methylene. Furthermore, methylene blue adsorption can be enhanced by employed the ultrasonic wave. This research aims to prepare, characterize, and employ blood clam shells as a low-cost adsorbent in the ultrasonic-assisted methylene blue adsorption. The parameters studied were the adsorbent preparation method, adsorbate pH, adsorbent mass, contact time, and initial concentration. The adsorbent preparation method affected the morphology, elemental composition, functional groups, and average particle size, which were confirmed using SEM-EDX, FTIR, and PSA, where this kind of investigation has never been conducted before. Optimum conditions for methylene blue adsorption were obtained for adsorbent A3, pH 12, adsorbent mass of 0.5 g, contact time of 60 min, and initial concentration of 10 ppm, with a removal percentage of 98.614 %. This removal percentage increased by 63.119 % (espescially in A3) compared to without ultrasonic waves, which could only remove methylene blue by 35.495 %. Ultrasonic-assisted adsorption of methylene blue using blood clam shells adsorbent proceeded according to the Sips isotherm model (R2 = 0.9953) and pseudo-second-order kinetics (R2 = 0.939). A3 shows an extraordinary performance, with reusability up to six cylces. The adsorption occurs by electrostatic interaction between adsorbent's surface and cations in methylene blue, chemisorption, and the formation of monolayer adsorbate in heterogenous A3's surface.

Evaluation of Tolerance and Selection of Heat-Tolerant Woody Plants against Heat Stress

Heat-tolerant species have become increasingly important because of climate change; however, a selection system for woody plants is not well established. This study was conducted to establish a selection system for heat-tolerant woody plant species and to select heat-tolerant species. After selecting heat-tolerant woody plants and applying heat stress to 27 species, the electrolyte leakage index (ELI) was measured. The ELI of the heat-tolerant species was lower than that of the non heat-tolerant species, and they survived well after heat stress treatment. For the preselected species, the degree of cell death was measured by Evans blue staining method, and the heat stress recovery ability was measured by a 3,3′-diaminobenzidine (DAB) staining method. The species showed less cell death even after heat treatment, and oxidative stress was low after recovery from heat stress. Traditional screening methods are mainly performed through field screening; however, this is difficult because it requires many samples and considerable time. The results of this study are relatively rapid, reproducible, and highly sensitive, so it is judged to be a method that can complement the existing traditional method as a heat-tolerant plant selection system. The results of this study can be widely used for the selection and breeding of heat-tolerant plants in preparation for climate change.

CX43 and oxidative stress are the targets of BCB staining to predict the developmental potential of buffalo oocytes.

This study used the brilliant cresyl blue (BCB) staining method to group buffalo oocytes (BCB+ and BCB-) and perform invitro maturation, invitro fertilization and embryo culture. At the same time, molecular biology techniques were used to detect gap junction protein expression and oxidative stress-related indicators to explore the molecular mechanism of BCB staining to predict oocyte developmental potential. The techniques of buffalo oocytes to analyse their developmental potential and used immunofluorescence staining to detect the expression level of CX43 protein, DCFH-DA probe staining to detect ROS levels and qPCR to detect the expression levels of the antioxidant-related genes SOD2 and GPX1. Our results showed that the invitro maturation rate, embryo cleavage rate and blastocyst rate of buffalo oocytes in the BCB+ group were significantly higher than those in the BCB- group and the control group (p < .05). The expression level of CX43 protein in the BCB+ group was higher than that in the BCB- group both before and after maturation (p < .05). The intensity of ROS in the BCB+ group was significantly lower than that in the BCB- group (p < .05), and the expression levels of the antioxidant-related genes SOD2 and GPX1 in the BCB+ group were significantly higher than those in the BCB- group (p < .05). Brilliant cresyl blue staining could effectively predict the developmental potential of buffalo oocytes. The results of BCB staining were positively correlated with the expression of gap junction protein and antioxidant-related genes and negatively correlated with the reactive oxygen species level, suggesting that the mechanism of BCB staining in predicting the developmental potential of buffalo oocytes might be closely related to antioxidant activity.

Agricultural low-cost waste adsorption of methylene blue and modelling linear isotherm method versus nonlinear prediction

AbstractThis study shows that geographically marked wheat hull, named Siyez, rice hull Sarı Kılçık, and Taşköprü Garlic stalk were used as agricultural waste to potential adsorbent materials for removing methylene blue from aqueous solution. Experimental data were evaluated in both equilibrium batch process and kinetic studies. In addition, the factors that affect the adsorption capacities, such as pH solutions, methylene blue concentration, contact time, and temperatures, were also investigated. Obtained data were subject to two constant adsorption models of Langmuir, Freundlich, Temkin, and Dubinin−Radushkevich. The kinetic models (pseudo-first-order, pseudo-second-order, intra-particle diffusion and film diffusion) and thermodynamic parameters were evaluated. The adsorption isotherms, characterized by an excellent fit with the Langmuir model (R2 = 0.99) across all adsorbents, underscore the prevalence of monolayer adsorption of methylene blue, in contrast to the Freundlich equation. Adsorption kinetics of the methylene blue onto the adsorbents followed pseudo-second-order kinetic model. According to high regression coefficient (R2) and minimal values of nonlinear error functions like RMSE; the maximum monolayer adsorption capacities of wheat hull, rice hull and garlic stalk were found to be 62.50 (mg/g), 54.94 (mg/g), and 370.37 (mg/g), respectively. The results indicated that these proposed adsorbents could be low-cost and effective adsorbents for water purification and have adsorption capacity as much as comparable with the literature. In batch equilibrium studies, the adsorption of methylene blue dye onto the wheat hull, rice hull, and garlic stalk exhibited a significant correlation with temperature, contact time, and initial concentration of methylene blue dye and Adaptive Neuro-Fuzzy Inference System algorithm for forecasting overall the system parameter well fitted with these findings with the accuracy of outputs (R2 about 0.99 for each). Consequently, the thermodynamic analysis revealed that the adsorption process takes place in bulk diffusion by liquid phase mass transfer and occurred spontaneously with endothermically except garlic stalk. Adsorption thermodynamic studies show that the adsorption of methylene blue onto the garlic stalk was spontaneous and exothermic. Graphical Abstract

Preosteoblast Adhesion and Viability Study of Freeze-Dried Bovine Bone Block Scaffold Coated with Human Umbilical Cord Mesenchymal Stem Cell Secretome.

Combining a three-dimensional scaffold with growth factors before implantation is one method used to increase scaffold bioactivity in bone tissue engineering. The mesenchymal stem cell (MSC)-conditioned medium (CM), called secretome, contains many proteins and growth factors required for tissue repair and growth. This study evaluated the bioactivity of a bovine bone scaffold combined with the secretome of human umbilical cord MSCs (hUC-MSCs) by analyzing MC3T3-E1 cell adhesion and viability on the scaffold. This in vitro laboratory study evaluated the effect of hUC-MSC secretome applied to bovine bone scaffolds processed using various techniques on MC3T3-E1 cell adhesion and viability. The three experimental groups included deproteinized bovine bone mineral-secretome (DBBM-CM), freeze-dried bovine bone-secretome (FDBB-CM), and decellularized FDBB-CM, whereas the control group was treated with DBBM alone. The cell adhesion test was performed using the centrifugation method after 6 and 24 hours, whereas the cell viability test was conducted using the trypan blue exclusion method after 24, 48, and 72 hours. Cell attachment was visualized after 4',6-diamidino-2-phenylindole staining and viewed under inverted fluorescence microscopy. Statistical analyses were performed using one-way analysis of variance, followed by a post hoc test in cases of significant differences. Statistical analyses showed significantly greater adhesion of the preosteoblasts to the FDBB-CM scaffold at 6 hours (p = 0.002). The results of the adhesion test at 24 hours and the viability tests at all observation times showed no significant differences (p > 0.05). This study found that the average MC3T3-E1 cell adhesions and viabilities were highest for the FDBB-CM and DBBM-CM scaffolds. DBBM scaffolds with the secretome had better cell adhesion and viability than those without the secretome. The addition of MSC secretome increased bovine bone scaffold bioactivity especially in DBBM and FDBB scaffolds.

Cu-doped 70S bioactive glass fibrous membranes produced using solution blow spinning (SBS)

3D fibrous membranes of undoped and Cu-doped 70S glass were developed, by solution blow spinning. The concentration of copper ions ranged from 1 to 4 % in mol. As-spun fibers were calcined/stabilized at 700 °C. The produced membranes' morphological and biological properties and the release of Ca and Cu ions were evaluated. Amorphous materials were obtained in pure and doped with 1 % Cu2+ 70S glass, while metallic copper was observed in 4 % Cu2+ doped glass. All samples have fibers with average diameter at the submicron scale, ranging from 700 to 850 nm. The amount of Cu ions influenced the degradation of the samples and the release of Cu and Ca ions in the Tris solution, the greater the amount of copper the more these ions were degraded and released in the solution. In vitro biological results, showed that adding copper favored the biomineralization process. No toxicity was observed in the MMT test for all samples. Alamar Blue method showed cell viability for fibroblasts greater than 90 % for all samples and for osteoblasts greater than 70 % for the BG-70S.2Cu sample. The developed fibrous membranes presented antimicrobial characteristics against Staphylococcus aureus and Candida albicans.