Ml Of Medium Research Articles

Development of e-Booklets making nata de pina as a learning resource for Biology Technology Innovation material in the Merdeka Phase E curriculum in high school

Research Development of e-Booklet Making Nata De Pina Research-Based Research as a Learning Source of Biological Technology Innovation Material in the Merdeka Phase E Curriculum in High School aims to analyze the influence of the concentration of sprouts extract on the quality of thickness and weight of nata de pina, as well as analyzing the validity, feasibility, and readability of E-Booklet Making Nata De Pina Developed. The development research design uses the Addie model, which consists of 5 stages, namely analysis, design, development, implementation and evaluation. In this study the implementation stage was not implemented. The results of this study were the provision of sprouts in the process of making nata de pina had a significant effect on the thickness of thickness and weight of nata de pina. The best results of the five treatments for the concentration of sprouts extract tested are trial in the K4 treatment with a concentration of sprouts of 100 ml for 500ml of media. The e-Booklet of making Nata de Pina is considered very valid with the acquisition of material validation scores of 90.90% and the validation score of media experts of 93.53%. The e-Booklet making Nata de Pina was declared very feasible with the acquisition of the teacher response score of 96.66% and the student response was 89.52%. As well as the results of the e-Booklet readability test making Nata de Pina is considered high with the acquisition of the Rumpang test score of 98.38%.

Growth of Litopenaeus vannamei using Synbiotics Supplementation Diet in Outdoor Low-Salinity Ponds Concerning Water Quality Parameters and Phytoplankton Communities

This study aimed to evaluate the influence of probiotic supplementation using L. bulgaricus FNCC-004 and prebiotics sodium alginate on the growth of Litopenaeus vannamei shrimp, as well as water quality parameters and plankton density in a five tons circle outdoor pond for 30 days of rearing. Assessment was conducted on specific pathogen-resistant (SPR) shrimp. Shrimp weight, plankton density, and water quality, with samples collected from circular ponds. The probiotic L. bulgaricus FNCC–004 was prepared as feed supplementation by adding 2% sodium alginate into the incubated bacterial media. One kg of feed was mixed with 200 mL of media containing the alginate-probiotic mixture. Feeding frequency was four times a day. Water quality sampling involved temperature, salinity, pH, dissolved oxygen (DO), and light intensity, measured twice daily. Plankton density was determined using a haemocytometer under microscope. Specific growth rate (SGR) of shrimp was calculated based on initial and final weights. Results showed variations in water quality parameters throughout the day, indicating the dynamic nature of the aquatic environment. Plankton density analysis revealed variations in species and abundance over the study period. The specific growth rate of Litopenaeus vannamei shrimp was recorded at 17.44% per day. This study enhances understanding of the effects of synbiotics supplementation in the diet on shrimp growth and environmental conditions in aquaculture systems

Production and Physicochemical Characterization of the Gel Obtained in the Fermentation Process of Blue Corn Flour (Zea mays L.) with Colletotrichum gloeosporioides.

Food gels are viscoelastic substances used in various gelled products manufactured around the world. Polysaccharides are the most common food gelling agents. The aim of this work was the production and characterization of a gel produced in a blue corn flour fermentation process, where different proportions were used of blue corn (Zea mays L.) flour and Czapek Dox culture medium (90 mL of culture medium with 10 g of blue corn flour, 80 mL of culture medium with 20 g of blue corn flour, and 70 mL of culture medium with 30 g of blue corn flour) and were fermented for three different durations (20, 25, and 30 days) with the Colletotrichum gloeosporioides fungus. A characterization of the gel was carried out studying the rheological properties, proximal analysis, toxicological analysis, microscopic structure, and molecular characterization, in addition to a solubility test with three different organic solvents (ethanol, hexane, and ethyl acetate, in addition to water). The results obtained showed in the rheological analysis that the gel could have resistance to high temperatures and a reversible behavior. The gel is soluble in polar solvents (ethanol and water). The main chemical components of the gel are carbohydrates, especially polysaccharides, and it was confirmed by FT-IR spectroscopy that the gel may be composed of pectin.

Biocompatibility Evaluation of Ampicillin-Loaded Whitlockite for Bone Regeneration.

Introduction Whitlockite (WH), a rare phosphate mineral within the apatite group, shows potential for bone regeneration owing to its superior composition and biocompatibility compared to hydroxyapatite. It can serve as a carrier for bioactive molecules, gradually releasing them to stimulate bone growth and expedite healing. This study aims to assess the biocompatibility of antibiotic-loaded WH, focusing on ampicillin, for bone regeneration applications. Methodology WH particles loaded with varying concentrations of ampicillin (10 and 25 mM) underwent biocompatibility assessments using the MTT assay. One gram of particles was incubated in 10 mL of culture medium for 24 and 48 hours. Experimental groups included control, WH, WH with ampicillin at 10 mM (WH+A10), WH with ampicillin at 25 mM (WH+A25), and positive control treated with 0.1% Triton X detergent. Subsequently, after a three-day culture period, RunX2 gene expression, indicative of osteoblastic differentiation, was quantified using real-time PCR analysis. Results Our research evaluated the bioactivity of WH particles treated with human osteoblastic cells using the MTT assay. While 10 mM ampicillin-loaded WH showed no significant difference in metabolic activity at both 24 and 48 hours, 25 mM ampicillin-loaded WH exhibited a slight reduction in metabolic activity at 24 hours, which normalized by 48 hours. Additionally, we assessed osteogenic potential and showed a significant increase in RunX2 expression with ampicillin-loaded WH, indicating sustained osteogenic properties. Conclusions Our study underscores the promising biocompatibility of WH particles by retaining their osteogenic properties even when, loaded with ampicillin, offering a potential avenue for future bone regeneration strategies.

Anabasis articulata (Forssk.) Moq. food aqueous extract triggers oxidative stress-induced senescence and reduces metastatic power in MDA-MB-231 cells

Ancient ethnobotanical practices handed down through traditional knowledge are still commonly employed to treat various pathologies, although the scientific reasons underlying their biological effects have not been clarified yet. In this contribution, the potential antitumoral activity of the aqueous extract from A. articulata (AAE) was investigated to validate the hypothesis of the Algerian folk medicine which would suggest this plant derivative as a functional food for treating breast cancer. A. articulata phytocomplex, isolated by maceration following exactly the African recipe, has been already characterized by our research group in previous works. Thus, the antiproliferative function of AAE against MDA-MB-231, a highly aggressive human breast adenocarcinoma cell line, was evaluated. Slowing down of cell growth, absence of cytotoxicity and DNA fragmentation, and cell cycle arrest at the G2/M phase were observed after treatment with AAE at different doses (0.3–6 mg of dried plant material equivalent per mL of culture medium) for 24 and 48 h. Wound and transwell assays proved that AAE possessed both antimigration and antiinvasive capacities, evidence also supported by molecular analyses focused on Metalloproteases (MMP-2 and MMP-9), Vimentin and ανβ3-Integrin. These results, together with the demonstration of the activation of p53/p21WAF1/Cip1/p27Kip1 pathway and the increase of oxygen reactive species levels, suggested that AAE triggered a senescence process. The final confirmation was obtained by a specific kit staining senescent cells. All our data would explain the efficacy of the Algerian medicinal remedy based on the intake of the investigated functional plant food and would highlight the basics for developing novel natural pharmacological products based on AAE and showing preventive and therapeutic antineoplastic potentialities against highly aggressive breast cancers.

Temporary immersion bioreactor as an efficient method for in vitro propagation of Agave marmorata

Agave marmorata is an important wild species in Mexico for mezcal and pulque production. Propagation by suckers and seeds is not sufficient to restore wild populations. The aim of this study was to develop a protocol for large-scale micropropagation of A. marmorata using a temporary immersion bioreactor (TIB). The effect of liquid medium using a TIB and semi-solid medium as the control treatment was compared. Different immersion frequencies and culture medium volumes per explant were evaluated in the TIB. In all treatments, the percentage of hyperhydricity, number of shoots per explant, shoot length, and total chlorophyll and carotenoid content were evaluated. In addition, the survival rate during acclimatization was evaluated. The results obtained show that the TIB with an immersion of 2 min every 8 h and with a culture medium volume of 50 mL had the best developmental parameters, with 19.60 shoots per explant, a shoot length of 1.74 cm and a percentage of hyperhydricity of 3.33 %. The highest chlorophyll content was observed in temporary immersion every 8 h, with 100, 75 and 50 mL of culture medium per explant. Regarding carotenoid content, the highest content was found at the 8 and 12 h immersion frequencies, with the culture medium volumes of 100, 75 and 50 mL per explant. For acclimatization, no differences were found in the survival rate between the culture systems evaluated. In conclusion, this study demonstrates that the use of a TIB is an alternative for large-scale micropropagation of A. marmorata.

Toxicity and cytokine release from human dental pulp stem cells after exposure to universal dental adhesives cured by single peak and polywave LEDs

Objectivesto assess the impact of universal adhesives, cured with single-peak and polywave LEDs, on the metabolic activity and cytokine release of human dental pulp stem cells (hDPSCs). In addition, analyze the degree of conversion (DC) of the adhesives cured with the different LEDs. MethodsDiscs (5 mm diameter, 1 mm thick) were prepared using three universal adhesives: Single Bond Universal (SBU, 3 M ESPE), Optibond Universal (OBU, Kerr), and Zipbond Universal (ZBU, SDI). These discs were cured for 40 s using a single-peak (DeepCure, 3 M ESPE) or a polywave light-emmiting diode (LED) curing unit (Valo Grand, Ultradent). After 24 h, the specimens were placed in 24-well culture plates, each containing 1 mL of culture medium for 24 h. hDPSCs (1.8 ×104) were seeded in 96-well plates and allowed to grow for 24 h. Subsequently, the cells were exposed to the extracts (culture medium containing eluates from the adhesive discs) for an additional 24 h. Cells not exposed to the extracts were used as a control group. The mitochondrial metabolism was assessed using the MTT assay and the cytokine release evaluated through MAGPIX. The degree of conversion of the adhesives was analyzed using FTIR (n = 5). The results were analyzed by ANOVA two-way and Tukey’s test. ResultsOBU and ZBU eluates caused a statistically significant reduction in mitochondrial metabolism, regardless of the LED used, indicating their cytotoxicity. In contrast, SBU did not significantly affect the MTT results, resembling the control group. A higher release of cytokines IL-1, IL-6, IL-10, and TNF-α were found in association to ZBU. SBU, on the other hand, increased the release of IL-8. OBU did not influenced the cytokine release. SBU presented the higher DC, while OBU and ZBU had similar DC, lower than SBU. SignificanceIn conclusion, universal adhesives exhibit toxicity towards hDPSCs, but the extent of toxicity varies depending on the adhesive material. ZBU was associated with increased cytokine release, particularly pro-inflammatory mediators, from hDPSCs. The different LEDs did not influenced the cytotoxicity of the evaluated adhesives.

Efficient and economic protein labeling for NMR in mammalian expression systems: Application to a preT-cell and T-cell receptor protein.

Protein nuclear magnetic resonance(NMR) spectroscopy relies on the ability to isotopically label polypeptides, which is achieved through heterologous expression in various host organisms. Most commonly, Escherichia coli is employed by leveraging isotopically substituted ammonium and glucose to uniformly label proteins with 15N and 13C, respectively. Moreover, E. coli can grow and express proteins in uniformly deuterium-substituted water (D2O), a strategy useful for experiments targeting high molecular weight proteins. Unfortunately, many proteins, particularly those requiring specific posttranslational modifications like disulfide bonding or glycosylation for proper folding and/or function, cannot be readily expressed in their functional forms using E. coli-based expression systems. One such class of proteins includes T-cell receptors and their related preT-cell receptors. In this study, we present an expression system for isotopic labeling of proteins using a nonadherent human embryonic kidney cell line, Expi293F, and a specially designed media. We demonstrate the application of this platform to the β subunit common to both receptors. In addition, we show that this expression system and media can be used to specifically label amino acids Phe, Ile, Val, and Leu in this system, utilizing an amino acid-specific labeling protocol that allows targeted incorporation at high efficiency without significant isotopic scrambling. We demonstrate that this system can also be used to express proteins with fluorinated amino acids. We were routinely able to obtain an NMR sample with a concentration of 200 μM from 30 mL of culture media, utilizing less than 20 mg of the labeled amino acids.

Nephroprotective effect of bone marrow mesenchymal stem cells on cisplatin induced kidney injury in albino rats

isplatin is a chemotherapeutic drug used in the treatment of a variety of cancers, with a known side effect of nephrotoxicity. Using stem cell therapy represents a distinctive and encouraging approach to remediate damaged organs. The administration of bone marrow mesenchymal stem cells (BMMSCs) has the potential to mitigate the adverse effects of cisplatin nephrotoxicity, thus helping with both functional and histological recuperation. Twenty-four mature male albino rats were divided into four groups. 1 ml of normal saline was injected intraperitoneally (I.P.) into the control group. Cisplatin was injected once (6 mg/kg I.P.) into the cisplatin group. 0.5 ml of culture media with 5 x 106 BMMSCs was injected i.p. with 6 mg/kg I.P. cisplatin in the BMMSC group. The withdrawal group received no treatment after cisplatin injections. At different times, groups were sacrificed. Kidney specimens were made for histology and immunohistochemistry. Morphometric and statistical studies were done. Blood urea and serum creatinine were evaluated before sacrifice. There were statistically significant differences between the studied groups regarding markers of incidence of acute tubular necrosis and recovery, suggesting that cisplatin therapy caused acute tubular necrosis, whereas BMMSCs improved renal function markers, including blood urea and serum creatinine levels and tissue restoration. Stem cell rats also showed cluster of differentiation 44 (CD44) in cells near tubules, helping injured kidneys regenerate tubular cells. The use of bone-marrow-derived mesenchymal stem cells (BMMSCs) mitigated the nephrotoxic effects of cisplatin, thus showing a restorative effect on both functional and histological parts.

Absorbed Bioactive Compounds Replicate Guanxin II-Induced Endothelium-Associated in/ex vivo Vasodilation.

To develop an interference-free and rapid method to elucidate Guanxin II (GX II)'s representative vasodilator absorbed bioactive compounds (ABCs) among enormous phytochemicals. The contents of ferulic acid, tanshinol, and hydroxysafflor yellow A (FTA) in GX II/rat serum after the oral administration of GX II (30 g/kg) were detected using ultra-performance liquid chromatography-mass spectrometry. Totally 18 rats were randomly assigned to the control group (0.9% normal saline), GX II (30 g/kg) and FTA (5, 28 and 77 mg/kg) by random number table method. Diastolic coronary flow velocity-time integral (VTI), i.e., coronary flow or coronary flow-mediated dilation (CFMD), and endothelium-intact vascular tension of isolated aortic rings were measured. After 12 h of exposure to blank medium or 0.5 mmol/L H2O2, endothelial cells (ECs) were treated with post-dose GX II of supernatant from deproteinized serum (PGSDS, 300 µL PGSDS per 1 mL of culture medium) or FTA (237, 1539, and 1510 mg/mL) for 10 min as control, H2O2, PGSDS and FTA groups. Nitric oxide (NO), vascular endothelial growth factor (VEGF), endothelin-1 (ET-1), superoxide dismutase (SOD), malondialdehyde (MDA) and phosphorylated phosphoinositide 3 kinase (p-PI3K), phosphorylated protein kinase B (p-AKT), phosphorylated endothelial nitric oxide synthase (p-eNOS) were analyzed. PGSDS was developed as a GX II proxy of ex vivo herbal crude extracts. PGSDS effectively eliminates false responses caused by crude GX II preparations. When doses equaled the contents in GX II/its post-dose serum, FTA accounted for 98.17% of GX II -added CFMD and 92.99% of PGSDS-reduced vascular tension. In ECs, FTA/PGSDS was found to have significant antioxidant (lower MDA and higher SOD, P<0.01) and endothelial function-protective (lower VEGF, ET-1, P<0.01) effects. The increases in aortic relaxation, endothelial NO levels and phosphorylated PI3K/Akt/eNOS protein induced by FTA/PGSDS were markedly abolished by NG-nitro-L-arginine methyl ester (L-NA, eNOS inhibitor) and wortmannin (PI3K/AKT inhibitor), respectively, indicating an endothelium-dependent vasodilation via the PI3K/AKT-eNOS pathway (P<0.01). This study provides a strategy for rapidly and precisely elucidating GX II's representative in/ex vivo cardioprotective absorbed bioactive compounds (ABCs)-FTA, suggesting its potential in advancing precision ethnomedicine.

Optimization of mineral nutrients to improve rhamnolipid production by Pseudomonas aeruginosa6 K‐11

AbstractBACKGROUNDRhamnolipids are biosurfactants that have their main application in bioremediation processes. They are able to increase the biodegradation and detoxification rates from industrial effluents. The north of Peru and Iquitos have been not only major oil producers but also have been responsible for oil spills. This study therefore focused on the optimization of mineral nutrients such as nitrogen, phosphorous, calcium and iron to improve rhamnolipids production by Pseudomonas aeruginosa 6 K‐11, which was isolated from oil soil in Peru.RESULTSPseudomonas aeruginosa 6 K‐11 was reactivated in Trypticase Soy Broth by scaling from 5 to 200 mL of culture medium for 8 h at each stage. The culture was centrifuged and placed in 100 mL Siegmund–Wagner medium for 36 h in a 500 mL Erlenmeyer flask. First, it was determined that sodium nitrate, monopotassium phosphate, calcium chloride and iron sulfate were the best sources of mineral nutrients. Then, by using corn oil as carbon source, the ratios obtained for carbon‐to‐nitrogen and carbon‐to‐phosphorous were 21.172 and 16.279, respectively. In addition, it was found that the optimal concentration for calcium was 0.046 g L−1 and for iron was 0.003 g L−1. The most influential variables were carbon‐to‐nitrogen and carbon‐to‐phosphorus ratio. Finally, 35.124 g L−1 rhamnolipids was produced after 168 h of fermentation with the optimal parameters.CONCLUSIONThe optimization of mineral nutrients was an important achieved goal for rhamnolipid production. In addition, this optimization can be used for modifying the culture medium to grow the biosurfactant producer strain. © 2024 Society of Chemical Industry (SCI).

Ядерно-цитоплазматическое созревание ооцитов Sus Scrofa Domesticus в условиях сверхнизких температур

Abstract. Vitrification of female gametes is the most important tool for solving the strategic tasks of cellular reproductive technologies in animal husbandry, veterinary medicine and biomedicine: replication of elite individuals (cloning, transgenesis), biodiversity conservation, organ xenotransplantation. The aim of this study was to evaluate the parameters of nuclear cytoplasmic maturation (chromatin status and mitochondrial activity) and developmental competence of donor porcine oocytes subjected to intra- (IOV) or extra-ovarian (EOV) vitrification and cultured in various maturation systems. Methods. Donor pig oocytes (6–8 months old) were vitrified extra- or intraovarially using cryoprotective agents (DMSO, ethylene glycol, trehalose, sucrose). The composition of cryoprotective agents was supplemented with 0.001 % highly dispersed silica nanoparticles (HDSns). Oocytes were thawed in trehalose solutions in TS-199 with 10 % of fetal bovine serum (FBS). Oocytes were cultured for 44 hours in media supplemented with granulosa cells (106 cells per ml of medium) and/or 0.001% of HDSns at 38.5 °C, in an atmosphere of 5 % CO2. Fertilization in vitro was carried out in accordance with the recommendations presented by us earlier [17. p. 17]. The functional activity of mitochondria and the status of chromatin in oocytes were assessed by double staining using MitoTracker Orange CMTMRos and Hoechst 33258 dyes. Results. The level of matured oocytes was 61 % at EOV and 29 % at IOV, and the level of cleavage embryos in EOV was in 11 % (31 % vs. 20 %, P &lt; 0.05) higher than at IOV when HDSns with granulosa cells were added in culture media. The maximum indicators of the functional activity of mitochondria among devitrified oocytes were in the group of EOV oocytes have cultured in media supplemented with HDSns and granulosa cells (152 ± 14.9 μA). Scientific novelty. The protocols for intra- and extra-ovarian vitrification of porcine oocytes have been modernized. Addition to cryoprotective agents and culture media of HDSns and co-culture of devitrified oocytes with granulosa cells significantly increased the fertility rates of devitrified gametes.

Application of cyanobacteria in radioactive waste

The extremely toxic radioactive wastes whose radioactivity persists for thousands of years are accumulated through nuclear power plants, mining companies, industries, and research centers, among others. However, the lack of technology for the treatment and removal of radioactivity from radioactive waste becomes a bottleneck for research. Currently, there are temporary solutions, and the radioactive waste continues to increase. Given this scenario it is necessary to seek alternatives for the treatment of radioactive waste with efficiency and low cost, without polluting the environment, and that is not harmful to human health. One way to reduce this type of pollution is to use microorganisms, especially cyanobacteria, which comprise one of the largest, most ecologically diverse, successful, and important group of bacteria on Earth. These bacteria have a great ability to remove pollutants, such as heavy metals, textile dyes, pesticides, etc., but their role in the degradation of recalcitrant and radioactive compounds is still scarce. The present work aimed to investigate the potential of cyanobacteria isolated from different environments to remove radiolabeled molecules from radioactive waste (containing 14C radioisotopes). Seven cyanobacteria, Synechococcus sp. CENA136 (isolated from the Mangrove of Cardoso Island), Phormidium autumnale UTEX 1580 (collection of UTEX cultures - fish aquarium), Nostoc sp CENA420 (Antarctica), Limnothrix sp. CENA458 (isolated from the reservoir), Oscillatoria acuminata CENA525 (isolated from the Pantanal), Nodularia sp CENA215 (isolated from Caatinga), and Trichormus SP UFV-56 (isolated from the Federal University of Viçosa-MG) were used in the present study. Cyanobacteria were maintained in Z8, BG-11, and SWBG-11 liquid culture medium under constant fluorescent lighting of 40 µmol photons·m-2·s-1 and a controlled temperature of 23 ± 1°C. Radioactive waste containing several 14C organic molecules, solubilized in organic solvents was used. The inoculums were obtained from 50 mL of culture medium. The radioactive waste was added at concentrations of 0, 5, 10 and 15%. To investigate the removal of the radioactivity present in the waste, analysis was performed by Liquid Scintillation Counting (LSC) in the beginning and final cultivation and HPLC (coupled to a flow scintillation analyzer) in order to evaluate the profile of radioactive molecules present in the waste. Intracellularly accumulated radioactivity was also assessed by LSC after cell disruption. The ability to accumulate radioactive molecules intracellularly was observed in all cyanobacteria, but Nostoc sp CENA420 accumulated 43% and Trichormus sp UFV-56 accumulated 68%, both cultivated with 15% of radioactive waste. These results showed that the cyanobacteria Nostoc sp CENA420 and Trichormus sp UFV-56 consumed 87 and 86% of radioactive molecules, respectively, being of great potential for the removal of radiolabeled radioactive waste molecules. Research using cyanobacteria to remove radiolabeled molecules from radioactive waste is still at an early stage but is a promising alternative to biological treatment.

Evaluation of organosulfur compound degradation property of a noble bacterial strain of Proteus mirabilis

Organosulfur compounds asssociated with coal are the major contributors to environmental pollution. An organosulfur compound dibenzothiophene (DBT) occupies about 70% of all the organosulfur compounds detected in coal. Microbial desulfurization processes are evolving as a tool for the remediation of organosulfur compounds. The present work deals with a microbial processing technique developed for the degradation of an organosulfur compound DBT by a noble bacterial strain Proteus mirabilis isolated from petroleum-contaminated soil. The study revealed that the bacterium has a high tolerance towards higher concentrations of DBT and performed a normal rate of bacterial growth in a culture medium supplemented with 50 mM of DBT. The Proteus mirabilis bacterium catalyzed the microbial desulfurization of DBT present in the culture medium. It was estimated that the bacterium desulfurized about 99% of the DBT incubated in 500 mL of culture medium at 37 °C ± 2 with the Proteus mirabilis for 5 days. The high-performance liquid chromatography (HPLC) analysis showed the presence of 2-hydroxybiphenyl in the bacterial culture medium indicating that the bacterium has followed a sulfur-specific 4S microbial pathway for the degradation of DBT through the production of 2-hydroxybiphenyl.

Reusable and Practical Biocomposite Based on Sphingopyxis sp. YF1 and Polyacrylonitrile-Based Carbon Fiber for the Efficient Bioremediation of Microcystin-LR-Contaminated Water.

Microbial degradation is a cost-effective and environmentally friendly method for removing microcystin-LR (MC-LR). However, the application of free bacteria has limitations due to low operational stability and difficulties in recovery. In a previous study, our group successfully isolated a highly efficient MC-LR-degrading bacterium, Sphingopyxis sp. YF1, from Taihu. To enhance its practical potential in addressing MC-LR-contaminated water pollution, a novel biological material named polyacrylonitrile-based carbon fiber @Sphingopyxis sp. YF1 (PAN-CF@YF1) was synthesized. The immobilization conditions of strain Sphingopyxis sp. YF1 on PAN-CF surfaces were optimized using Box-Behnken design and response surface methodology (RSM), which turned out to be an optimal pH of 7.6 for the culture medium, a ratio of 0.038 g of supporting materials per 100 mL of culture media, and an incubation time of 53.4 h. The resultant PAN-CF@YF1 showed a great degradation effect both for low and high concentrations of MC-LR and exhibited satisfactory cyclic stability (85.75% after six cycles). Moreover, the application of PAN-CF@YF1 in the bioreactors demonstrated effective and sustainable MC-LR removal, with a removal efficiency of 78.83% after three consecutive treatments. Therefore, PAN-CF@YF1 with high degradation activity, environmental compatibility, straightforward preparation, and recyclability shows significant application potential for the bioremediation of MC-LR-contaminated water bodies.

Evaluation of the production of oppositional cytokines IL-6 and IL-10 in the culture of mononuclear blood leukocytes in rats with metabolic syndrome

Aim: to evaluate the secretory activity of mononuclear blood leukocytes by the level of in vitro production of oppositional cytokines (pro-inflammatory IL-6 and anti-inflammatory IL-10) in rats in a model of diet-induced metabolic syndrome (MS) against the background of changes in the concentration of adipokines leptin and adiponectin.&#x0D; Materials and methods. In male Wistar rats, MS was modeled by keeping for 12 weeks on a high-fat and high-carbohydrate diet; control group animals received a normal diet. Mononuclear leukocytes isolated by gradient centrifugation from heparinized blood were cultured in a medium based on RPMI-1640 (concentration of 2×106 cells in 1 ml of medium) for 24 hours in sterile tubes in a CO2-incubator in two variants – without the addition of an inducer and with the addition of bacterial lipopolysaccharide (LPS). The concentration of cytokines in the culture fluid, hormones leptin and adiponectin in the blood serum was determined by enzyme immunoassay. Statistical data processing was carried out in the SPSS Statistics 23.&#x0D; Results. In animals exposed to a 12-week high-fat and high-carbohydrate diet, against the background of an increase in body weight, the specific mass of visceral adipose tissue, metabolic disorders (hyperglycemia, impaired glucose tolerance, dyslipidemia), high concentrations of leptin and low concentrations of adiponectin, the ability of mononuclear blood leukocytes to produce IL-6 and IL-10 in vitro at the basal level and when stimulated by bacterial LPS does not change.&#x0D; Conclusion. Probably, the morphofunctional status and secretory profile of monocyte-macrophage system cells depends on the severity and duration of the metabolic and hormonal imbalance associated with obesity and systemic chronic inflammation.

HISTOLOGICAL STRUCTURE OF THE RAT FEMURS AFTER FILLING OF DEFECTS IN THE DISTAL METAPHYSIS WITH 3D-PRINTED IMPLANTS BASED ON POLYLACTIDE AND TRICALCIUM PHOSPHATE IN COMBINATION WITH MESENCHYMAL STROMAL CELLS

Polylactide (PLA) frameworks printed on a 3D printer are used for filling the bone defects. The osteotropic properties of 3D-PLA can be improved by combining with tricalcium phosphate (TCP) and mesenchymal stromal cells (MSCs). Objective. Study the reconstruction in the rat femurs after implanting 3D-printed implants based on PLA and TCP (3D-I) in combination with cultured allogeneic MSCs into defects in the distal metaphysis. Methods. 48 white laboratory rats (age 5–6 months) were used, which were randomly divided into groups: Control — 3D-I; Experiment-I — 3D-I, saturated MSCs; Experiment II — 3D-I, with injection of 0.1‒0.2 ml of medium with MSCs into the area of surgical intervention 7 days after implantation. 15, 30 and 90 days after the operation, histological (with histomorphometry) studies were conducted. Results. The area of 3D-I decreased with time in all groups and connective and bone tissues formed in different ratios. 15 days after the surgery, in the Experiment-I group, the area of the connective tissue was 1.9 and 1.6 times greater (p&lt;0.001) in comparison to the Control and Experiment II; 30 days it was greater 1.6 times (p &lt; 0.001) and 1.4 times (p=0.001), respectively. 30 days after the surgery, the area of newly formed bone in the Experiment-I group was 2.2 times (p &lt; 0.001) less than in the Control. On the contrary, in the Experiment-II, the area of newly formed bone was 1.5 and 3.3 times greater (p &lt; 0.001) compared to Experiment-I and Control, respectively. Conclusions. The studied 3D-I with time after their implantation into the metaphyseal defects of the rats’ femurs are replaced by connective and bone tissues. The use of 3D-I, saturated MSCs, 15 and 30 days after the surgery, caused excessive formation of connective tissue and slower bone formation. Local injection of MSCs 7 days after the implantation of 3D-I caused to the formation of a larger area of newly bone 30th day after surgery compared to 3D-I alone and 3D-I with MSCs.

Giant Duckweed (Spirodela polyrhiza) Root Growth as a Simple and Sensitive Indicator of Copper and Chromium Contamination.

Aquatic environment are often contaminated with heavy metals from various industrial sources. However, physicochemical techniques for pollutant detection are limited, thus prompting the need for additional bioassays. We investigated the use of greater duckweed (Spirodela polyrhiza) as a bioindicator of metal pollution. We exposed S. polyrhiza to four pollutants (namely, silver, cadmium, copper, and chromium) and assessed metal toxicity by measuring its frond area and the length of its regrown roots. The plant displayed significant differences in both frond size and root growth in response to the four metals. Silver was the most toxic (EC50 = 23 µg L-1) while copper the least (EC50 = 365-607 µg L-1). Direct comparisons of metal sensitivity and the reliability of the two endpoint assays showed that root growth was more sensitive (lower in terms of 50% effective concentration) to chromium, cadmium, and copper, and was more reliable (lower in terms of coefficient of variation) than those for frond area. Compared to conventional Lemna-based tests, the S. polyrhiza test is easier to perform (requiring only one 24-well plate, 3 mL of medium and a 72-h exposure). Moreover, it does not require livestock cultivation/maintenance, making it more suitable for repeated measurements. Measurements of S. polyrhiza root length may be suitable for assessment when copper and chromium in municipal and industrial wastewater exceed the environmentally permissible levels.

Micropropagation of iraca palm (Carludovica palmata Ruiz y Pav) using a temporary immersion system

Carludovica palmata is a Neotropical plant with a promising potential for the agroindustry. It is an important source of fibers used to manufacture handmade goods. Colombia is one of the leading countries in the production of this species; however, it lacks efficient techniques for its propagation. We developed a protocol for massive micropropagation of C. palmata using a temporary immersion bioreactor (TIB) system. Immersion frequency, immersion time, culture medium volume, and explant density were evaluated using a split-split-plot design. The variables evaluated were the number and length of shoots and roots, the number of leaves, and the dry weight of shoots. The performance of three micropropagation systems, TIB, semi-solid medium, and liquid medium, was evaluated using a generalized randomized block design. Murashige and Skoog (MS) culture medium with 1.0 mg L−1 of 6-benzylaminopurine, 0.5 mg L−1 of 1-naphthaleneacetic acid, and 20 g L−1 of sucrose added to it was used. The results obtained show a frequency of 12 h with an immersion time of 1 min improved the length of shoots, roots, and number of leaves, and dry weight. A volume of 20 mL/explant was found to be an adequate condition to increase the number and length of shoots, the number of leaves, and the dry weight. According to the optimization module, the factor levels that will maximize the evaluated variables are a 1-minute immersion of the tissues in the culture medium with a frequency of 12 hours each time, with 175 mL of culture medium per bioreactor and density of 10 explants. In conclusion, TIB was shown to be efficient for massive micropropagation of C. palmata compared with conventional methods.

P-116 How does the short insemination method affect clinical outcomes and laboratory management?

Abstract Study question Does the short insemination method using a time-lapse monitoring system improve clinical outcomes and laboratory management? Summary answer The method increased the normal fertilization rate of Conventional IVF and also increased the 3PN rate. The working environment of the laboratory staff was improved. What is known already Short insemination using a time-lapse monitoring system has been reported to reduce fertilization confirmation failures due to loss of pronuclei, but many reports indicate that the method was implemented to perform so-called 'rescue ICSI' for unfertilized oocytes on the day of egg retrieval and there are not many reports of clinical outcomes in conventional IVF without rescue ICSI. There are also few reports on the working environment and working hours in the laboratory. Study design, size, duration This was a retrospective study of 3,714 oocytes from 715 cycles in 595 patients who underwent Conventional IVF with egg retrieval in 2016-2022. Long insemination (1,577 oocytes) was performed in cycles up to May 2019 and short insemination (2,137 oocytes) in later cycles. In IVF, 40,000 motile sperm were placed in 1 ml of culture medium per oocyte. Participants/materials, setting, methods Oocytes of the long insemination group (group L) were inseminated 5 hours after egg retrieval and denuded 19 hours later to observe their pronuclei. Oocytes from the short insemination group (group S) were inseminated 2 hours after egg retrieval, denuded 4 hours later, and cultured in EmbryoScope (Vitrolife, Sweden) to observe the pronuclei. Since the working hours set at our clinic are 8:00 a.m.-5:00 p.m., the time schedule for both groups followed this schedule. Main results and the role of chance The 2PN rate was 60.9% in group L and 64.7% in group S. The 3PN rates were 7.5% and 10.2% respectively, both rates being significantly higher in group S (P &amp;lt; 0.05). The non-fertilization rates were significantly higher in group L (25.3% and 21.7% respectively); the 1PN rate was not significantly different (3.6% vs. 3.4%). The rate of missed pronuclei was 2.8% in group L and 0% in group S. Early embryo transfer pregnancy rate (25.4% vs 27.8%), good blastocyst development rate (52.2% vs 63.6%) and blastocyst transfer pregnancy rate (41.1% vs 31.7%) in groups L and S respectively. The good blastocyst incidence was significantly higher in group S. In the case of the Conventional IVF cycle in which egg retrieval was performed at 8:00 a.m., the embryologists in group L were to observe the pronuclei at 8:00 a.m. the next day, but in reality the embryologists had to come to work earlier because of denudation, and even then pronuclear loss occurred. In contrast, the group S did not require an earlier attendance because denudation was not performed the next morning, and the time-lapse images did not fail to confirm the pronuclei at all. Limitations, reasons for caution Since the short insemination method was introduced at our clinic in June 2019, this study compared clinical outcomes before and after that date. Wider implications of the findings It has been reported in the past that early insemination time increases both 2PN and 3PN rates. The short insemination method in this study also used a faster insemination time, which may have led to similar insemination results. Trial registration number not applicable