Culture Time Research Articles

ETIOPATHOGENETIC REASONING OF URETHRAL MICROBIOTA REHABILITATION IN PATIENTS WITH ACUTE INFECTIOUS URETHRITIS

The microbiota of the male genitourinary tract is a complex dynamic system that has not been sufficiently studied. Unlike the microbiome of the female urethral mucosa, which is constantly changing, the taxonomic composition and population level of the male urethral microbiome is more stable and independent of age. The aim of the study. To improve the results of treatment of patients with inflammatory processes of the urethra by determining the taxonomic composition, population level and microscopic indicators of the "macroorganism-microbiome" ecosystem of the microbiota of the urethra in men with acute urethritis.Material and methods. The taxonomic composition and population level of the first portion of urine of 69 sexually mature men (average age 34.8±7.14 years) with acute infectious (nonspecific) urethritis were investigated. Isolation of conditionally pathogenic bacteria was carried out by the bacteriological method using optimal nutrient media, their pH, temperature and cultivation time for each taxon. The identification of isolated pure cultures was carried out according to morphological, tinctorial, cultural, biochemical properties, as well as signs of pathogenicity and antigenic structure. To determine the mechanisms of colonization of the mucous membrane of the urethra by conditionally pathogenic microorganisms, the microecological method was used.Research results. According to the taxonomic composition and the level of microecological indicators of the "macroorganism-microbiome" ecosystem of the urethra microbiota (Margalef's species richness index, Whittaker's species diversity index, and Simpson's and Berger-Parker's species dominance indices), vulgar (common), hemolytic, and enteropathogenic Escherichia coli, coagulase-positive staphylococcus. The leading role in the formation of urethritis is played by ordinary enterobacteria - E. coli, a slightly smaller etiological role in the formation of urethritis in men belongs to S. aureus (less by 17.83%), E. coli Hly+ (less by 28.35%), enteropathogenic Escherichia coli (by 83.12%). Other microorganisms have a minimal etiological role. Conclusions. Acute urethritis in men is a polyetiological disease. Enterobacteria E. coli, E. coli Hly+, enteropathogenic Escherichia coli, K. pneumoniae, S. aureus, S. epidermidis, S. viridans are involved in the formation of the inflammatory process of the urethra. The leading causative agents of acute urethritis in men are E. coli, S. aureus, E. coli Hly+, enteropathogenic escherichia, which belong to uropathogenic escherichia.

Patient-derived bladder cancer organoids show stable transcript expression along cultivation.

Bladder cancer (BC) is a prevalent malignancy with high recurrence rates. Patient-derived bladder cancer organoids (BCO) pose as a promising approach in both, disease modeling and individualized treatment screening. The aim of this study was to investigate the transcriptomic plasticity in BCOs as a function of cultivation times to define ideal time periods for the applications envisioned. Tumor samples of three patients with pathologically confirmed non-muscle invasive and muscle-invasive bladder cancer were included in this study and expanded as BCOs. RNA expression was investigated at different time periods of cells in culture using differential gene expression for overall transcript expression and quantitative real-time PCR (qRT-PCR) for pathological relevant markers. Differential gene expression of the BCO lines was investigated across passages 1-4, in passages 5-9 and above 9, respectively. Analysis of the entire transcriptome of the respective BCO lines revealed consistent profiles without significant alterations throughout the cultivation and expansion procedure. Notably, key transcripts like TP53, PIK3CA, BRCA1, among others, exhibited stable expression levels in the quantitative RNA analysis during the cultivation period. The robust transcriptome during BCO cultivation advocates for the use of earlier passages of BCOs in personalized medicine providing a time-efficient drug screening option to accelerate the counseling of patients' treatment options. Higher passages of BCOs still hold the potential in topics demanding for expanded cell masses such as medical device development and others.

Formation of cell spheroids using Standing Surface Acoustic Wave (SSAW).

3D bioprinting becomes one of the popular approaches in the tissue engineering. In this emerging application, bioink is crucial for fabrication and functionality of constructed tissue. The use of cell spheroids as bioink can enhance the cell-cell interaction and subsequently the growth and differentiation of cells in the 3D printed construct with the minimum amount of other biomaterials. However, the conventional methods of preparing the cell spheroids have several limitations, such as long culture time, low-throughput, and medium modification. In this study, the formation of cell spheroids by SSAW was evaluated both numerically and experimentally in order to overcome the aforementioned limitations. The effects of excitation frequencies on the cell accumulation time, diameter of the formed cell spheroids, and subsequently, the growth and viability of cell spheroids in the culture medium over time were studied. Using the high-frequency (23.8 MHz) excitation, cell accumulation time to the pressure nodes could be reduced in comparison to that of the low-frequency (10.4 MHz) excitation, but in a smaller spheroid size. SSAW excitation at both frequencies does not affect the cell viability up to 7 days, > 90% with no statistical difference compared with the control group. In summary, SSAW can effectively prepare the cell spheroids as bioink for the future 3D bioprinting and various biotechnology applications (e.g., pharmaceutical drug screening and tissue engineering).

Adsorption of extracellular lipase in a packed-bed reactor: an alternative immobilization approach.

In light of the growing demand for novel biocatalysts and enzyme production methods, this study aimed to evaluate the potential of Aspergillus tubingensis for producing lipase under submerged culture investigating the influence of culture time and inducer treatment. Moreover, this study also investigated conditions for the immobilization of A. tubingensis lipase by physical adsorption on styrene-divinylbenzene beads (Diaion HP-20), for these conditions to be applied to an alternative immobilization system with a packed-bed reactor. Furthermore, A. tubingensis lipase and its immobilized derivative were characterized in terms of their optimal ranges of pH and temperature. A. tubingensis was shown to be a good producer of lipase, obviating the need for inducer addition. The enzyme extract had a hydrolytic activity of 23 U mL-1 and achieved better performance in the pH range of 7.5 to 9.0 and in the temperature range of 20 to 50°C. The proposed immobilization system was effective, yielding an immobilized derivative with enhanced hydrolytic activity (35 U g-1), optimum activity over a broader pH range (5.6 to 8.4), and increased tolerance to high temperatures (40 to 60 ℃). This research represents a first step toward lipase production from A. tubingensis under a submerged culture and the development of an alternative immobilization system with a packed-bed reactor. The proposed system holds promise for saving time and resources in future industrial applications.

Human research islet cell culture outcomes at the Alberta Diabetes Institute IsletCore

ABSTRACT Human islets from deceased organ donors have made important contributions to our understanding of pancreatic endocrine function and continue to be an important resource for research studies aimed at understanding, treating, and preventing diabetes. Understanding the impacts of isolation and culture upon the yield of human islets for research is important for planning research studies and islet distribution to distant laboratories. Here, we examine islet isolation and cell culture outcomes at the Alberta Diabetes Institute (ADI) IsletCore (n = 197). Research-focused isolations typically have a lower yield of islet equivalents (IEQ), with a median of 252,876 IEQ, but a higher purity (median 85%) than clinically focused isolations before culture. The median recovery of IEQs after culture was 75%, suggesting some loss. This was associated with a shift toward smaller islet particles, indicating possible islet fragmentation, and occurred within 24 h with no further loss after longer periods of culture (up to 136 h). No overall change in stimulation index as a measure of islet function was seen with culture time. These findings were replicated in a representative cohort of clinical islet preparations from the Clinical Islet Transplant Program at the University of Alberta. Thus, loss of islets occurs within 24 h of isolation, and there is no further impact of extended culture prior to islet distribution for research.

Machine Learning Assisted MALDI Mass Spectrometry for Rapid Antimicrobial Resistance Prediction in Clinicals.

Antimicrobial susceptibility testing (AST) plays a critical role in assessing the resistance of individual microbial isolates and determining appropriate antimicrobial therapeutics in a timely manner. However, conventional AST normally takes up to 72 h for obtaining the results. In healthcare facilities, the global distribution of vancomycin-resistant Enterococcus fecium (VRE) infections underscores the importance of rapidly determining VRE isolates. Here, we developed an integrated antimicrobial resistance (AMR) screening strategy by combining matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) with machine learning to rapidly predict VRE from clinical samples. Over 400 VRE and vancomycin-susceptible E. faecium (VSE) isolates were analyzed using MALDI-MS at different culture times, and a comprehensive dataset comprising 2388 mass spectra was generated. Algorithms including the support vector machine (SVM), SVM with L1-norm, logistic regression, and multilayer perceptron (MLP) were utilized to train the classification model. Validation on a panel of clinical samples (external patients) resulted in a prediction accuracy of 78.07%, 80.26%, 78.95%, and 80.54% for each algorithm, respectively, all with an AUROC above 0.80. Furthermore, a total of 33 mass regions were recognized as influential features and elucidated, contributing to the differences between VRE and VSE through the Shapley value and accuracy, while tandem mass spectrometry was employed to identify the specific peaks among them. Certain ribosomal proteins, such as A0A133N352 and R2Q455, were tentatively identified. Overall, the integration of machine learning with MALDI-MS has enabled the rapid determination of bacterial antibiotic resistance, greatly expediting the usage of appropriate antibiotics.

Development of a PCR-based method to identify fetal sex during IVF cycles.

One of the most recognizable cases of preimplantation genetic diagnosis (PGD) is X-linked diseases. Diagnosis of fetal sex is essential for couples who are known to be at risk of some X-linked disorders. The objective of this study was to discriminate between female (XX) and male (XY) embryos by detecting sex chromosomes-specific sequences in spent culture medium and comparing these results to PGD/CGH array results. It may open new window for the development of a non-invasive PGD method. 120 Embryo's spent media from Day 3 and Day 5 embryos were collected. Modified phenol-chloroform solution was used for DNA extraction from spent media. Sex determination was performed using SRY, TSPY and AMELOGENIN evaluation through quantitative polymerase chain reaction (q-PCR) method. IBM SPSS and MedCalc were used for statistical analyses to compare sex determination of embryos by spent medium with PGD/CGH array results. Culture time was demonstrated to increase the DNA amount among day 5 embryos culture medium samples. Non-invasive PGD by means of spent culture medium gave a sensitivity, specificity, positive predictive value and negative predictive value of 100% for sex determination. Results of sex determination using spent medium by q-PCR were consistent with the results of PGD/CGH array. Improvements in cell-free DNA extraction and PCR amplification procedures provide us an effective method to perform a PGD test without biopsy in the future, especially about X-linked diseases.

Anti-Inflammatory Effect of Medicinal Fungus Antrodia cinnamomea Cultivated on Pinus morrisonicola Hayata.

The fungus Antrodia cinnamomea, which grows on Cinnamomum kanehirae tree, has many medicinal uses. However, its cultivation using the traditional method of growing on the C. kanehirae tree is costly and time-consuming. A possible alternative method of cultivating A. cinnamomea is to use Pinus morrisonicola Hayata tree, as it contains α-terpineol, which stimulates the synthesis of triterpenoids. To compare the cultivation of A. cinnamomea on P. morrisonicola and C. kanehirae, the contents of triterpenoids and antcin were determined using high-performance liquid chromatography. Anti-inflammatory effects of the extracts of each product were investigated in lipopolysaccharide (LPS)-stimulated BV-2 cells. Their mechanisms on mitogen-activated protein kinase (MAPK) signalling pathways (p38, c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK)) were determined using Western blot analysis. The results showed that the cultivation times of A. cinnamomea on P. morrisonicola and traditional C. kanehirae discs were drastically different, lasting 6 and 18 months, respectively. The concentration of triterpenoids in the corresponding fruiting bodies was (70.0±3.0) and (20.0±4.0) mg/mL, respectively. More antcins were produced in the P. morrisonicola culture. Similar anti-inflammatory effect was obtained by both cultures, which is confirmed by the reduced production of IL-1β, IL-6, COX-2 and nitrogen monoxide. Their mechanisms were confirmed by the suppression of MAPK signalling pathways. Cultivation on P. morrisonicola is an innovative and more cost-effective method for growing A. cinnamomea. The same anti-inflammatory effect is achieved in a shorter production time.

Longitudinal tracking of perfluorooctanoic acid exposure on mammary epithelial cell spheroids by dynamic optical coherence tomography.

We investigated the morphology and intracellular motility of mammary epithelial cell (MCF10DCIS.com) spheroids cultured in 3D artificial extracellular matrix under perfluorooctanoic acid (PFOA) exposure. Dynamic optical coherence tomography (OCT) was employed for real-time, non-invasive imaging of these spheroids longitudinally over 12 days under PFOA exposures up to 500 µM. Despite no significant changes in volume or asphericity of spheroids, morphological alterations were observed in OCT images of spheroids at 100 µM on Day 12 and from Day 4 at 500 µM. Intracellular motility was assessed by the inverse-power-law exponent of the speckle fluctuation spectrum (α), and an autocorrelation-based motility amplitude (M). Linear regression indicated that both PFOA concentration and culture time are highly significant predictors for both α and M (p < 0.001 for all). Both PFOA concentration and culture time have positive associations with α and negative association with M, where increased α indicates suppression of higher frequency fluctuations (∼> 2 Hz) relative to those at lower frequencies, and decreased M indicates overall suppression of intracellular motility. This study can lead to the future development of biomarkers for per- and polyfluoroalkyl substances (PFAS) exposure using dynamic OCT and its associated toolkit of quantitative metrics.

Isolation, culture and identification of brain tumor stem cells from glioma

Objectives: This study aims to provide a feasible method for culturing tumor stem cells to facilitate further research on the occurrence and development of brain tumors, making it applicable for in vitro studies of brain tumor stem cells. Methods: Samples of glioma were collected and subsequently isolated and cultured, then immunocytochemical staining, including CD133, Nestin, and GFAP staining, was performed on the cultured cells. Results: Some of the primary cultured tumor stem cells grew in the shape of sphere and formed into clusters. With the extension of culture time, the cell mass gradually increased, with strong proliferation and self-renewal ability. Meanwhile, the tumor derived spherical cells could differentiate into different types of tumor cells in serum-containing medium since most of them were positive to CD133, Nestin, and GFAP on the 7 days. Conclusion: The tumor stem cell can differentiate into different cell types, carrying significant implications for tumor research and treatment. More importantly, the methods introduced in this study are simple and feasible to isolate and culture tumor stem cell, contributing to cell source for future studies.

Optimal Planting Time for Summer Tomatoes (Lycopersicon esculentum Mill.) Cropping in Korea: Growth, Yield, and Photosynthetic Efficiency in a Semi-Closed Greenhouse.

In Korea, greenhouses are traditionally used for crop cultivation in the winter. However, due to diverse consumer demands, climate change, and advancements in agricultural technology, more farms are aiming for year-round production. Nonetheless, summer cropping poses challenges such as high temperatures, humidity from the monsoon season, and low light conditions, which make it difficult to grow crops. Therefore, this study aimed to determine the best planting time for summer tomato cultivation in a Korean semi-closed greenhouse that can be both air-conditioned and heated. The experiment was conducted in the Advanced Digital Greenhouse, built by the National Institute of Agricultural Sciences. The tomato seedlings were planted in April, May, and June 2022. Growth parameters such as stem diameter, flowering position, stem growth rate, and leaf shape index were measured, and harvesting was carried out once or twice weekly per treatment from 65 days to 265 days after planting. The light use efficiency and yield per unit area at each planting time was measured. Tomatoes planted in April showed a maximum of 42.9% higher light use efficiency for fruit production and a maximum of 33.3% higher yield. Furthermore, the growth form of the crops was closest to the reproductive growth type. Therefore, among April, May, and June, April is considered the most suitable planting time for summer cultivation, which is expected to contribute to reducing labor costs due to decreased workload and increasing farm income through increased yields. Future research should explore optimizing greenhouse microclimates and developing crop varieties tailored for summer cultivation to further enhance productivity and sustainability in year-round agricultural practices.

Enzyme-Free In-Situ Electrochemical Measurement Using a Porous Membrane Electrode for Glucose Transport into Cell Spheroids.

Microphysiological systems have attracted attention because of their use in drug screening. However, it is challenging to measure cell functions in real time using a device. In this study, we developed a cell culture device using a porous membrane electrode for in situ electrochemical glucose measurements for cell analysis. First, a porous membrane electrode was fabricated and electrochemically evaluated for enzyme-free glucose measurement. Subsequently, the glucose uptake of MCF-7 spheroids was evaluated using living spheroids, fixed spheroids, supernatants, and glucose transporter inhibitor-treated spheroids. Conventionally, the direct optical measurement of glucose uptake requires fluorescence-labeled glucose derivatives. In addition, the glucose uptake can be evaluated by measuring the glucose concentration in the medium by optical or electrochemical measurements. However, glucose needs to be consumed in the entire cell culture medium, which needs a long culture time. In contrast, our system can measure glucose in approximately 5 min without any labels because of in situ electrochemical measurements. This system can be used for in situ measurements in in vitro cell culture systems, including organ-on-a-chip for drug screening.

Development of New Multi-Glycosylation Routes to Facilitate the Biosynthesis of Sweetener Mogrosides from Bitter Immature Siraitia Grosvenorii Using Engineered Escherichia coli.

Mogrosides, which have various pharmacological activities, are mainly extracted from Siraitia grosvenorii (Luo Han Guo) and are widely used as natural zero-calorie sweeteners. Unfortunately, the difficult cultivation and long maturation time of Luo Han Guo have contributed to a shortage of mogrosides. To overcome this obstacle, we developed a highly efficient biosynthetic method using engineered Escherichia coli to synthesize sweet mogrosides from bitter mogrosides. Three UDP-glycosyltransferase (UGT) genes with primary/branched glycosylation catalytic activity at the C3/C24 sites of mogrosides were screened and tested. Mutant M3, which could catalyze the glycosylation of nine types of mogrosides, was obtained through enhanced catalytic activity. This improvement in β-(1,6)-glycosidic bond formation was achieved through single nucleotide polymorphisms and direct evolution, guided by 3D structural analysis. A new multienzyme system combining three UGTs and UDP-glucose (UDPG) regeneration was developed to avoid the use of expensive UDPG. Finally, the content of sweet mogrosides in the immature Luo Han Guo extract increased significantly from 57% to 95%. This study not only established a new multienzyme system for the highly efficient production of sweet mogrosides from immature Luo Han Guo but also provided a guideline for the high-value utilization of rich bitter mogrosides from agricultural waste and residues.

Growth analysis of six lisianthus (Eustoma grandiflorum) cultivars in different growing seasons

The objective of work was to evaluate the growth of six lisianthus cultivars at different times, in a protected environment, in the southern region of Brazil. The experimental design was randomized blocks, in split plots, with three replications. The plots were constituted by the lisianthus cultivars: 'White Excalibur', 'Blue Picotee', 'Blue Echo', 'Arena Red', 'Robella' and 'Echo Champagne'and the subplots by the dates of plant collections (evaluations), which were carried out at the time of transplantation (0) and 21, 42, 63 and 84 days after transplantation (DAT) for seasons 1(October 2017 to January 2018), 2 (November 2017 to February 2018) and 5 (August to December 2018); and at (0) and at 30, 60, 90, 120, 150 and 180 DAT at seasons 3 (April to November 2018), 4 (July to December 2018) and 6 (February to June 2019). All cultivars showed an increasing tendency to accumulate a dry mass of the plants until the harvest of the floral stems. The maximum dry mass value was obtained by the cultivar 'Robella' (20.3 g/plant-1) in season 3. For all cultivars, the absolute and relative growth rates of the crop increased with the advancement of the cultivation period. In seasons 1, 2 and 5 all cultivars showed higher growth velocities between 63 and 84 days after transplanting (DAT). In seasons 3 and 4 the cultivars showed higher growth from 90 to 150 DAT. In seasons 1 and 2 the cultivars showed a higher rate of plant growth, resulting in a shortening of the cycle. The cultivars conducted in seasons 3 and 4 had a longer cultivation time due to the autumn/winter period. The cultivar 'Arena Red' showed the lowest growth rates in all seasons.

Evaluation of Gelatin-Based Poly(Ester Urethane Urea) Electrospun Fibers Using Human Mesenchymal and Neural Stem Cells.

Previously, a new biodegradable poly(ester urethane urea) was synthesized based on polycaprolactone-diol and fish gelatin (PU-Gel). In this work, the potential of this new material for neural tissue engineering is evaluated. Membranes with randomly oriented fibers and with aligned fibers are produced using electrospinning and characterized regarding their mechanical behavior under both dry and wet conditions. Wet samples exhibit a lower Young's modulus than dry ones and aligned membranes are stiffer and more brittle than those randomly oriented. Cyclic tensile tests are conducted and high values for recovery ratio and resilience are obtained. Both membranes exhibited a hydrophobic surface, measured by the water contact angle (WCA). Human mesenchymal stem cells from umbilical cord tissue (UC-MSCs) and human neural stem cells (NSCs) are seeded on both types of membranes, which support their adhesion and proliferation. Cells stained for the cytoskeleton and nucleus in membranes with aligned fibers display an elongated morphology following the alignment direction. As the culture time increased, higher cell viability is obtained on randomfibers for UC-MSCs while no differences are observed for NSCs. The membranes support neuronal differentiation of NSCs, as evidenced by markers for a neuronal filament protein (NF70) and for a microtubule-associated protein (MAP2).

Epidemiology and outcomes of bacterial meningitis in the neonatal intensive care unit.

Examine pathogen distribution, antibiotic resistance patterns, and hospital outcomes of infants with bacterial meningitis in neonatal intensive care units (NICUs) in the US from 2013-2018. Infants were divided into 2 groups based on age at the time of meningitis: early-onset (0-3 days) and late-onset (>3 days). We compared demographics, clinical characteristics, epidemiology, hospital outcomes, distribution of organisms and resistance, and blood culture timing relative to cerebrospinal fluid culture. From 345 NICUs, 659 infants were diagnosed with bacterial meningitis. The cumulative incidence was 1.1-1.3 cases/1000 NICU discharges. Median gestational age was 33 weeks, median birth weight was 1910 grams, 12% failed hearing screening, and 9% died prior to discharge. Of 141 cases of E. coli meningitis, 53% were resistant to ampicillin. Significant morbidities occur in infants with culture-proven meningitis in NICUs. Culture and subsequent discernment of sensitivity are crucial to guide definitive therapy.

Glucosamine sulfate-loaded nanofiber reinforced carboxymethyl chitosan sponge for articular cartilage restoration

Cartilage is severely limited in self-repair after damage, and tissue engineering scaffold transplantation is considered the most promising strategy for cartilage regeneration. However, scaffolds without cells and growth factors, which can effectively avoid long cell culture times, high risk of infection, and susceptibility to contamination, remain scarce. Hence, we developed a cell- and growth factor-dual free hierarchically structured nanofibrous sponge to mimic the extracellular matrix, in which the encapsulated core–shell nanofibers served both as mechanical supports and as long-lasting carriers for bioactive biomass molecules (glucosamine sulfate). Under the protection of the nanofibers in this designed sponge, glucosamine sulfate could be released continuously for at least 30 days, which significantly accelerated the repair of cartilage tissue in a rat cartilage defect model. Moreover, the nanofibrous sponge based on carboxymethyl chitosan as the framework could effectively fill irregular cartilage defects, adapt to the dynamic changes during cartilage movement, and maintain almost 100 % elasticity even after multiple compression cycles. This strategy, which combines fiber freeze-shaping technology with a controlled-release method for encapsulating bioactivity, allows for the assembly of porous bionic scaffolds with hierarchical nanofiber structure, providing a novel and safe approach to tissue repair.

Wastewater treatment using a microalgae consortium mainly composed of chlorella, in Mexico

The bioremediation capacity of microalgae found in the wastewater channels in Irrigation District 03 (DR03) in Mexico was studied. The study evaluated the effect of pH, aeration, light intensity, and water source on the productivity, culture time, and composition of the microalgal biomass, predominantly made up of Chlorella vulgaris spp. Parameters such as Chemical Oxygen Demand (COD), total solids (TS), and turbidity were monitored to assess the efficiency and bioremediation capacity of the microbial consortium. The yield production of microalgal biomass was measured to evaluate the amount produced per unit volume of treated water. Two culture conditions were assessed: uncontrolled and controlled conditions of aeration and lighting. Under controlled culture conditions, a productivity of 0.031 g L−1 d−1 biomass with 17% lipidic fraction and 37.65% ash content was observed, along with a reduction in COD and turbidity by 57 and 85%, respectively. A biomass productivity of 0.024 g L−1 d−1 was obtained.

Study on Optimization of Liquid Fermentation Medium and Antitumor Activity of the Mycelium on Phyllopora lonicerae.

Phylloporia lonicerae is an annual fungus that specifically parasitizes living Lonicera plants, offering significant potential for developing new resource food and medicine. However, wild resources and mycelium production of this fungus is limited, and its anti-tumor active ingredients and mechanisms remain unclear, hampering the development of this fungus. Thus, we optimized the fermentation medium of P. lonicerae and studied the anti-tumor activity of its mycelium. The results indicated that the optimum fermentation medium consisted of 2% sucrose, 0.2% peptone, 0.1% KH2PO4, 0.05% MgSO4·7H2O, 0.16% Lonicera japonica petals, 0.18% P fungal elicitor, and 0.21% L. japonica stem. The biomass reached 7.82 ± 0.41 g/l after 15 days of cultivation in the optimized medium, a 142% increase compared with the potato dextrose broth medium, with a 64% reduction in cultivation time. The intracellular alcohol extract had a higher inhibitory effect on A549 and Eca-109 cells than the intracellular water extract, with half-maximal inhibitory concentration values of 2.42 and 2.92 mg/ml, respectively. Graded extraction of the alcohol extract yielded petroleum ether phase, chloroform phase, ethyl acetate phase, and n-butanol phase. Among them, the petroleum ether phase exhibited a better effect than the positive control, with a half-maximal inhibitory concentration of 113.3 μg/ml. Flow cytometry analysis indicated that petroleum ether components could induce apoptosis of Eca-109 cells, suggesting that this extracted component can be utilized as an anticancer agent in functional foods. This study offers valuable technical support and a theoretical foundation for promoting the comprehensive development and efficient utilization of P. lonicerae.

Investigation of the Use of Mycelial Filler with Different Cultivation Times for the Filtration of Particulate Airflow

Balanced and sustainable development has challenges in utilizing the best efficiency technologies, using new types of materials with reduced environmental impact, including composite types, reusable materials, and easily recyclable consumer adaptable products. One understudied biologically produced material is mycelium, the scientifically studied and improved cultivation of which produces an environmentally friendly material with unique properties with a wide range of applications. In this work, filtration fillers from mycelia of different cultivation periods and their abilities to filter airflow from solid particles were experimentally studied. Numerical modeling studied the interaction and trapping of particles in the flow with the surface of mycelium filters. The results of the research revealed a high airflow filtration efficiency of more than 91%, as well as differences and advantages in the properties and structure of mycelia of different growth periods, and the need for further study of this biomaterial.