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Nanobubbles (NBs) are a technology applicable in various sectors and have proven to be able to accelerate plant germination and growth and increase agricultural yields. The use of NBs in tissue culture still needs to be done. Tissue culture is a technology that can produce a large number of seeds in a short time especially for orchid plants. NBs in orchid tissue culture are expected to accelerate orchid growth. This research aims to identify the growth of Protocorm Like Bodies (PLB) of Dendrobium sp. orchids induced by NBsO2, and to determine the effects of NBsO2 on the growth of Dendrobium sp. orchids. The research method involves various steps such as sterilization, making pre-treatment media, making liquid media, sterilizing tools and materials, sterilizing nanogenerators, making NBsO2 solutions, screening NBs, pre-treatment, induction of Nanobubbles, and measuring plants. The observation parameters include PLB weight (g), PLB height (cm), number of living PLB (%), PLB color, root emergence, root length, shoot emergence, number of new shoots, embryo emergence, and number of embryos. The results showed that the growth of Dendrobium sp. orchid PLB induced by NBsO2 experienced a rapid and significant growth rate compared to treatments that were not induced by NBsO2. The use of NBsO2 affects the growth of Dendrobium sp. orchids by increasing the time of embryo emergence, number of embryos, plant weight, plant height, color, and percentage of life to increase the accelerated growth rate of Dendrobium sp. orchid PLB. [ABSTRACT FROM AUTHOR]

The optimization of tissue culture protocols for blueberry propagation is crucial to meet the increasing demand for disease-free planting materials. This study aimed to determine the effects of three auxins (IBA, IAA, and NAA) at different concentrations to enhance root and shoot development of two blueberry cultivars with different abilities to root, 'Lanmei #1'and 'Brightwell'. Results showed that NAA at 0.5 mg·L−1 significantly promoted shoot number and length in 'Lanmei #1' while 'Brightwell' showed a limited response. IAA had a variable impact on both shoot and root parameters, with 'Lanmei #1' showing more pronounced positive responses. IBA showed a differential response on root and shoot parameters in both cultivars, with 'Lanmei #1' showing a superior response for shoot number, length, root length, and overall plant vigor. Activated carbon had minimal effect on growth parameters across both cultivars. 'Lanmei #1' demonstrated a robust response to auxin treatments, suggesting it is more suitable for in vitro rooting. The present results confirm previous data on the different abilities of blueberry genotypes to root and the necessity of optimizing the rooting protocol depending on the cultivar. [ABSTRACT FROM AUTHOR]

The disadvantages of practical date palm micropropagation are tissue browning, low callus proliferation rate, low multiplication efficiency, and vitrification. The aim of the study was to determine the effect of ancymidol (Ancy) and phloroglucinol (PG) on the growth and some biochemical components of the 'Barhee' date palm cultured in vitro. The combination of 0.75 mg·l−1 Ancy and 50 mg·l−1 PG was found to be the most effective in terms of callus regeneration rate (89%) and number of shoots (14.3). A reduction in browning was observed in tissues cultured on media supplemented with 0.75 mg·l−1 Ancy in combination with 25 or 50 mg·l−1 PG. The medium supplemented with 0.75 mg·l−1 Ancy and 50 mg·l−1 PG eliminated shoot vitrification. Effective micropropagation was associated with increased carbohydrate and protein content. In this study, the genetic stability of plants obtained by micropropagation was confirmed by DNA-based RAPD fingerprinting. The results may indicate that the micropropagation protocol used in this study was suitable and applicable to the production of genetically stable date palm plants on a mass scale. [ABSTRACT FROM AUTHOR]

The biocompatibility of dental materials is critical for ensuring safety in clinical applications. However, standard in vitro cytotoxicity assays often rely on stiff tissue culture plastic (TCP), which does not accurately replicate the biomechanical properties of soft oral tissues. In this study, we compared human gingival fibroblasts (HGFs) cultured on soft, gel-based substrates mimicking gingival tissue stiffness (0.2 kPa) with those cultured on conventional TCP (3 GPa) to assess the influence of substrate stiffness on the cytotoxicity of methyl methacrylate (MMA), as well as other cytotoxic agents, including DMSO and H2O2. The results demonstrated that cells cultured on softer substrates exhibited enhanced resistance to cytotoxic stress, with increased viability and decreased apoptosis and DNA damage following exposure to MMA, DMSO, and H2O2. Notably, HGFs on soft substrates showed significantly greater resilience to MMA-induced cytotoxicity compared to those cultured on TCP. These findings emphasize the critical role of substrate stiffness in modulating cellular responses to toxic agents and highlight the necessity of using physiologically relevant models for cytotoxicity testing of dental materials. This study provides valuable insights for improving biocompatibility assessment protocols in clinical settings. [ABSTRACT FROM AUTHOR]

Background: Evaluation of infectious virus titer is a challenge for respiratory syncytial virus (RSV) clinical trials because of the labile nature of RSV and rapid loss of infectivity without proper specimen handling. However, there has been no rigorous investigation into RSV stability in clinical specimens. Methods: RSV stability was investigated by evaluating virus titers and defined as titer variation from baseline within three standard deviations of our titration assay. RSV stability in viral transport medium (VTM) at 4°C and the effect of freezing method on stability were evaluated using RSV‐A2 stock. RSV stability in nasal aspirates collected in VTM at 4°C was estimated by regression analysis of virus titers measured at several time points. Stability of these specimens stored at −80°C for 10–15 months after freezing by the method, which maintained RSV‐A2 stability, was also assessed. Results: Three standard deviations were calculated from our titration assay as 0.97 log10 50% tissue culture infectious dose (TCID50/mL), and RSV stability was defined as variation of virus titer from baseline within 1.0 log10TCID50/mL. RSV‐A2 in VTM at 4°C was stable for at least 120 h. Freezing at −80°C negatively affected virus stability, whereas freezing in liquid nitrogen or a dry ice‐ethanol bath did not. RSV in nasal aspirates was stable for 2 days at 4°C and for 10–15 months at −80°C after snap freezing. Conclusions: RSV in nasal aspirates in VTM was estimated to be stable for 2 days at 4°C and for approximately 1 year at −80°C. [ABSTRACT FROM AUTHOR]

Eucheuma denticulatum and Kappaphycus alvarezii are two of the most commercially important, cultivated carrageenophytes in Southeast Asia. Their large-scale farming through repeated vegetative propagation and virtual monocropping has resulted in a variety of phyconomic issues. The efficacy of two European, commercial liquid extracts from the brown seaweeds, i.e., Ascophyllum nodosum (ANE), a fucoid, and Laminaria digitata (LDE), a kelp, on the direct formation of axes and survival of these two eucheumatoids was examined using tissue culture techniques. Seaweed apical sections (2–3 mm long) were cultured for 45 days, with different concentrations of ANE and LDE (i.e., 0.005, 0.05, 0.5, 1.5, and 2.5 mL L−1) and compared with a control (UV-filtered seawater). Lower concentrations (< 1.5 mL L−1) of both liquid extracts promoted morphogenesis and growth of E. denticulatum and K. alvarezii in tissue culture, with the dilution rate of 0.05 mL LDE L−1 resulting in a high percentage survival (i.e., 95.3 ± 0.9%), formation of direct axes (i.e., 95.3 ± 0.9%), and the longest length (i.e., 10.0 ± 0.3 mm) for E. denticulatum after a typical 45-day culture period. A 0.5 mL ANE L−1 concentration was found to be beneficial for the tissue culture of K. alvarezii, given the relatively high final survival and direct axes formation (i.e., 96.2 ± 2.2%) and axes length (i.e., 8.2 ± 0.1 mm). This study supports the application of these temperate, brown seaweed-derived extracts as a phycobiostimulant enrichment in eucheumatoid micropropagation for mass production of plantlets for out-planting purposes. [ABSTRACT FROM AUTHOR]

Several studies have been conducted in the last 3 years to evaluate tilapia's parasitic and bacterial infection in cultures in Mexico. The present work evaluates the severity, prevalence, and health condition through the histopathological index based on the damage in the organs and tissues of tilapia cultured in ponds and cages during 2018 and 2019. Seven hundred and eight fish were collected from ponds and cages of tilapia farms in Chiapas, Oaxaca, and Guerrero and preserved and processed for histological observations. The prevalence of histopathological alterations found in each biological system was calculated and expressed in percentage. The histopathological index of each fish (HIfish) was calculated based on two different semi-quantitative methods. Histologically, the tilapia showed alterations as inflammation, melanomacrophage centers, hyperplasia, telangiectasias, and epitheliocystis unrelated to specific infectious pathogenic agents in both years but suggest effects associated with diverse factors in different biological systems. The prevalence was higher in all the parameters evaluated in 2018. The pathological manifestations in both years show that the health condition of the fish was affected regardless of the type of culture (cages and ponds), with the respiratory and digestive systems the most affected. These results show a baseline of the health condition of the tilapias cultured in southeast Mexico. The observed damages were not lethal but reflected a constant exposition to stressful conditions to trigger chronic inflammatory responses in the fish and make them susceptible to pathogens present in the environment. [ABSTRACT FROM AUTHOR]

Biomanufacturing enables novel sources of compounds with constant demand, such as food coloring and preservatives, as well as new compounds with peak demand, such as diagnostics and vaccines. The COVID-19 pandemic has highlighted the need for alternative sources of research materials, thrusting research on diversification of biomanufacturing platforms. Here, we show initial results exploring the walnut somatic embryogenic system expressing the recombinant receptor binding domain (RBD) and ectodomain of the spike protein (Spike) from the SARS-CoV-2 virus. Stably transformed walnut embryo lines were selected and propagated in vitro. Both recombinant proteins were detected at 3–14 µg/g dry weight of tissue culture material. Although higher yields of recombinant protein have been obtained using more conventional biomanufacturing platforms, we also report on the production of the red pigment betanin in somatic embryos, reaching yields of 650 mg/g, even higher than red beet Beta vulgaris. This first iteration shows the potential of biomanufacturing using somatic walnut embryos that can now be further optimized for different applications sourcing specialized proteins and metabolites. [ABSTRACT FROM AUTHOR]

In vitro micropropagation protocols for Hylocereus undatus and Hylocereus polyrhizus pitaya species were developed in Murashige and Skoog (MS) and Lloyd and McCown (WPM) nutrient media. During micropropagation stage, 0, 3, 4, 5, 6 mgL−1 Benzyl adenine (BA), 0.2 mgL−1 Gibberellic acid (GA3), and 0.1 mgL−1 Indole butyric acid (IBA) were added to the MS and WPM media. For the rooting stage, 0, 0.5, 1, 1.5, 2 mgL−1 IBA, 0.5 mgL−1 Naphthalene acetic acid (NAA), 0.2 mgL−1 GA3 and 0.1 mgL−1 BA were added to the same media. H. undatus reached the highest number of shoots (8.94) in the MS medium containing 3 mgL−1 BA while H. polyrhizus achieved the highest number of shoots (4.46) in the MS medium containing 5 mgL−1 BA. The WPM medium (3.08) produced a higher average number of roots compared to the MS medium (2.55). The highest average root number was obtained from 0.5 mgL−1 IBA concentration, followed by 2 mgL−1 IBA concentration. H. polyrhizus (51.63 mm) had a higher average root length than H. undatus (49.59 mm). The MS medium (51.22 mm) was more effective in promoting average root length compared to the WPM medium (49.99 mm). The highest average shoot fresh weight (1183.33 mg) and shoot dry weight (106.00 mg) were obtained from H. undatus in the WPM medium containing 2.0 mgL−1 IBA. H. polyrhizus (399.97 mg) had a higher average root fresh weight than H. undatus (275.60 mg) with the 2 mgL−1 of IBA concentration yielding the highest average value (604.83 mg). H. polyrhizus (35.93 mg), the WPM medium (38.30 mg), and 2 mgL−1 of IBA concentration (49.75 mg) resulted in a higher average root dry weight. [ABSTRACT FROM AUTHOR]

The genetic richness of the Capsicum genus holds significant potential for breeding programs, but it hinders the integration of biotechnological approaches for genetic improvement. In vitro manipulation and regeneration of pepper species have been limited by the inherent recalcitrance barriers of the genus, which mainly manifest in the massive obtention of structures with multiple shoot apical meristem (SAM) deformations. Fifty years of pepper tissue culture have demonstrated that explant responsiveness to culture conditions is essentially governed by molecular and physiological factors, and it is highly genotype-dependent. The empirical approach of the research carried out in this field has allowed to substantially improve the in vitro response by modifying the surrounding environment. Currently, there are several reports of pepper regeneration through organogenesis, somatic embryogenesis, and androgenesis. However, these protocols are quite inefficient and little reproducible in other Capsicum genotypes. Literature about pepper tissue culture has extensively reviewed the plant regeneration success achieved, but little is known about the multifactorial nature of this persistent phenomenon. This article summarizes the progress made in understanding the recalcitrant nature of the Capsicum genus, highlighting the influence of the molecular and physiological processes involved in the in vitro response. [ABSTRACT FROM AUTHOR]

Copper oxide nanoparticles (CuONPs) offer promising antimicrobial properties against a range of pathogens, addressing the urgent issue of antibiotic resistance. This study details the synthesis of glutamic acid-coated CuONPs (GA-CuONPs) and their functionalisation on medical-grade silicone tubing, using an oxysilane bonding agent. The resulting coating shows significant antimicrobial activity against both Gram-positive and Gram-negative bacteria, including multidrug-resistant strains, while remaining non-toxic to human cells and exhibiting stable adherence, without leaching. This versatile coating method can be applied during manufacturing, or for ad hoc modifications, enhancing the antimicrobial properties of medical devices. [ABSTRACT FROM AUTHOR]

Biomarkers for cancer immunotherapy are an unmet medical need. The group of Daniela Thommen at the NKI recently reported on novel methodologies based on short-term cultures of patient-derived tumor fragments whose cytokine concentrations in the supernatants and activation markers on infiltrating T cells were associated with clinical response to PD-1 blockade. We set up a similar culture technology with tumor-derived fragments using mouse tumors transplanted into syngeneic immunocompetent mice to test an agonist anti-CD137 mAb and its combinations with anti-PD-1 and/or anti-TGF-β. Increases in IFNγ concentrations in the tissue culture supernatants were detected upon in-culture activation with the anti-CD137 and anti-PD-1 mAb combinations or concanavalin A as a positive control. No other cytokine from a wide array was informative of stimulation with these mAbs. Interestingly, increases in Ki67 and other activation markers were substantiated in lymphocytes from cell suspensions gathered at the end of 72 h cultures. In mice bearing bilateral tumors in which one was excised prior to in vivo anti-CD137 + anti-PD-1 treatment to perform the fragment culture evaluation, no association was found between IFNγ production from the fragments and the in vivo therapeutic outcome in the non-resected contralateral tumors. The experimental system permitted freezing and thawing of the fragments with similar functional outcomes. Using a series of patient-derived tumor fragments from excised solid malignancies, we showed IFNγ production in a fraction of the studied cases, that was conserved in frozen/thawed fragments. The small tumor fragment culture technique seems suitable to preclinically explore immunotherapy combinations. [ABSTRACT FROM AUTHOR]

Di(2-ethhylhexyl) phthalate (DEHP) is a common plastic rubberizer. DEHP leaches from plastic matrices and is under increasing scrutiny as numerous studies have linked it to negative human health manifestations. Coxsackievirus B3 (CVB) is a human pathogen that typically causes subclinical infections but can sometimes cause severe diseases such as pancreatitis, myocarditis, and meningoencephalitis. Though CVB infections are common, severe illness is relatively rare, and it is unclear what factors mediate disease severity. In this study, we sought to determine the effects that DEHP has on CVB infection in a variety of human cell types to evaluate whether this plastic-derived pollutant could represent a proviral environmental factor. Methods: HeLa cervical cancer cells, human induced pluripotent stem cell-derived brain-like endothelial cells (iBECs), and Caco-2 colon carcinoma cells were exposed to 40 µg/mL DEHP for 24 h prior to infecting with enhanced green fluorescent protein (EGFP)-expressing CVB. The severity of the infection was evaluated via fluorescence microscopy and flow cytometry-based viral EGFP detection, viral plaque assay on tissue culture media, and Western blotting to detect VP1 viral capsid protein. Interferon-associated proteins such as interferon regulatory factor (IRF) 3, IRF7, interferon-induced transmembrane (IFITM) 2, and IFITM3 were measured by Western blotting. The roles of IFITM2 and IFITM3 in the context of CVB infection were evaluated via siRNA silencing. Results: We found that DEHP drastically increased CVB infection in each of the cell types we tested, and, while the cellular processes underlying DEHP's proviral properties were not entirely clear, we observed that DEHP may subvert CVB-induced interferon signaling and elevate levels of IFITMs, which appeared to bolster CVB infection. Conclusions: DEHP may represent a major environmental factor associated with the severity of CVB infection. Further understanding of how DEHP exacerbates infection may better elucidate its potential role as a proviral environmental factor. [ABSTRACT FROM AUTHOR]

Objectives This study aims to evaluate the effects of Croton grewioides essential oil (CGEO) and anethole on follicle survival, growth, and oxidative stress in cultured bovine ovarian tissues. Methods Ovarian tissues were cultured for 6 days in a medium supplemented with different concentrations (1, 10, 100, or 1000 µg mL–1) of CGEO or anethole and then, follicular survival and growth, collagen content, and stromal cell density in ovarian tissues cultured in vitro were evaluated by histology. The mRNA levels of superoxide dismutase (SOD) , catalase (CAT) , glutathione peroxidase 1 (GPX1) , peroxirredoxin 6 (PRDX6), and nuclear factor erythroid 2-related factor 2 (NRF2) were evaluated by real-time PCR. The activity of SOD, CAT, glutathione peroxidase (GPx), and thiol concentrations were investigated. Key findings Ovarian tissues cultured with 1 µg mL–1 CGEO or anethole had a higher percentage of healthy follicles than those cultured in a control medium (P <.05). The 1 µg mL–1 CGEO also increased the number of stromal cells, collagen fibers, and thiol levels. Anethole (1 µg mL–1) increased CAT activity and reduced that of GPx. The activity of SOD was reduced by CGEO. In contrast, 1 µg mL–1 anethole reduced mRNA for CAT, PRDX1, and NRF2 (P <.05). In addition, 1 µg mL–1 CGEO reduced mRNA for CAT , PRDX6 , and GPx1 (P <.05). Conclusions The presence of 1 µg mL–1 anethole or CGEO in a culture medium promotes follicle survival and regulates oxidative stress and the expression of mRNA and activity of antioxidant enzymes in cultured bovine ovarian tissues. [ABSTRACT FROM AUTHOR]

The patchouli plant (Pogostemon cablin Benth.) is a tropical herbaceous plant that produces essential oil. One of the problems is that the production is not yet optimal. Fulfillment of superior seedlings can help increase patchouli productivity. Conventional patchouli propagation through stem cuttings is ineffective and takes longer. Patchouli propagation can be done using a tissue culture approach via direct organogenesis to produce seedlings quickly and efficiently. Effective patchouli propagation methods and successful acclimatization are very important to research to support the propagation and breeding of patchouli plants. The aim of this research was to determine the best of BAP concentration in direct organogenesis of leaf and stem explants. The research design used a completely randomized series of hormone BAP, it has 5 levels, namely 0 mg/L (as control), 0.25 mg/L, 0.50 mg/L, 0.75 mg/L, and 1.0 mg/L. The explants used were the leaves and stems of Aceh patchouli. Plantlets are acclimatized in compost media and covering treatment. Based on the results of observations, the best BAP concentration is 0.25 mg/L with the initial observation parameters of the early emergence of shoots, number of shoots, and length of shoots on leaf explants were 10 daps, 35.33 shoots, and 2.83 cm respectively. The use of leaf explants showed a better response compared to stem explants. Patchouli plantlets were successfully acclimatized and can adapt to the ex vitro environment using the covering method. Successful patchouli propagation and high acclimatization can help produce effective patchouli seeds. [ABSTRACT FROM AUTHOR]

In order to investigate the reproduction of seedless, seeded and ornamental barberry shrubs through tissue culture, a factorial experiment in the form of a completely randomized design in three replications and with three factors, the first factor of three types of barberry shrubs (seedless, seeded and ornamental), the second factor of BAP hormone at four levels (0, 1, 2, 3 mg l-1) and the third factor of IBA hormone at two levels (0 and 0.25 mg l-1) was done in the laboratory of Islamic Azad University, Damghan branch in 2019. The culture medium used was based on Murashige and Stooge (MS). After preparing the culture medium and sterilizing it by autoclave, it was distributed in the test tubes. After preparation, the samples were cultured in a sterile environment and then the samples were stimulated to grow using growth hormones. The investigated traits included germination, number of leaves, length, width and surface of leaves, ratio of width to length of leaves, fresh weight and dry weight of leaves. After collecting information, the data obtained from the experiment was organized and then analysis of variance was performed with SAS statistical software and comparison of means with Duncan's multi-range test. Pearson's correlation coefficient was used in order to determine the relationships of traits. The main effects and some interaction effects on the time of greening, number of leaves, length, width, leaf area and leaf width to length ratio, leaf fresh weight and dry weight were significant. The correlation coefficient shown between the dry weight of leaves with the number of leaves (r=0.88**), leaf length (r=0.74**), leaf width (r=0.74**), leaf area (r=0.75**), green leaf weight(r=0.96**) positive correlation was significant at 1% probability level. [ABSTRACT FROM AUTHOR]

Adipose tissue plays a crucial role in metabolic syndrome, autoimmune diseases, and many cancers. Because of adipose's role in so many aspects of human health, there is a critical need for in vitro models that replicate adipose architecture and function. Traditional monolayer models, despite their convenience, are limited, showing heterogeneity and functional differences compared to 3D models. While monolayer cultures struggle with detachment and inefficient differentiation, healthy adipocytes in 3D culture accumulate large lipid droplets, secrete adiponectin, and produce low levels of inflammatory cytokines. The shift from monolayer models to more complex 3D models aims to better replicate the physiology of healthy adipose tissue in culture. This study introduces a simple and accessible protocol for generating adipose organoids using a scaffold-free spheroid model. The method, utilizing either 96-well spheroid plates or agarose micromolds, demonstrates increased throughput, uniformity, and ease of handling compared to previous techniques. This protocol allows for diverse applications, including drug testing, toxin screening, tissue engineering, and co-culturing. The choice between the two methods depends on the experimental goals, with the 96-well plate providing individualized control and the micromold offering scale advantages. The outlined protocol covers isolation, expansion, and characterization of stromal vascular fraction cells, followed by detailed steps for spheroid formation and optional downstream analyses. [ABSTRACT FROM AUTHOR]

The purpose of the research was to improve the morpho-anatomical and physiological features of somatic embryos (SEs) of date palms in tissue culture media that have the potential to produce green and healthy plantlets. Copper sulfate (CuSO4) and cobalt chloride (CoCl2) were supplemented to date palm maturation medium at various concentrations, separately or combined. Adding CoCl2 to the media of maturation produced the greatest relative growth and differentiation percentages of date palm embryogenic calli, particularly at 0.52 mgL-1. In comparison to the control, when combined treatments were administered alone, the quantity of the produced somatic embryos rose dramatically (17/jar). Total sugars and free amino acids were the highest in SEs grown on a maturation medium with CoCl2 at 0.26 mgL-1. Moreover, total phenols and indoles were picked significantly at their higher levels in the generated SEs that were cultured on a medium enhanced with 0.52 mgL-1. When the maturation medium was administered along with both as a treatment, the antioxidant enzyme, ascorbate peroxidase, reached its maximum activity. Whereas, adding 0.52 mgL-1 of CoCl2 to the culture medium enhanced the activities up to of highest levels of ascorbate peroxidase, polyphenol oxidase and phenylalanine ammonia-lyase. [ABSTRACT FROM AUTHOR]

This case report describes the orthodontic management of a case of chronic non-bacterial osteomyelitis (CNO) of the mandible, a rare non-infective variant of osteomyelitis that exhibits a marked predilection in children and adolescents. The patient presented with a unilateral facial swelling associated with fluctuating pain. Radiographic examination, along with tissue biopsy and culture, as well as multispecialty input, led to confirmation of the diagnosis. There was no clear aetiological factor and pharmacological, symptomatic management was indicated. CNO requires multidisciplinary input, with good interspecialty communication and discussion, for an accurate diagnosis. Orthodontic management should be considered on a case-by-case basis, with tailored aims appropriate for each patient. [ABSTRACT FROM AUTHOR]

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Jojoba is an important tropical oil crop, and jojoba oil is widely used in the aerospace lubricant and cosmetic industries. Jojoba exhibits high tolerance to droughts and high temperatures. However, there is currently a lack of rapid and effective methods for identifying stress-tolerant genes in jojoba. Here, an efficient hairy root genetic transformation system of jojoba (Simmondisa chinensis) mediated by Agrobacterium rhizogenes was established and used for the functional evaluation of ScGolS1, a putative stress-tolerant gene. First, using the leaves of jojoba as explants, transgenic jojoba hairy roots carrying the RUBY gene were obtained under sterile conditions using the "soaking co-cultivation method". Second, we optimized the four conditions affecting hairy root genetic transformations, namely, the strains of A. rhizogenes, co-cultivation under light or dark conditions, the infection time, and the OD600 value of the bacterial suspension. The following best transformation conditions were determined, A. rhizogenes K599, light during co-cultivation, an infection time of 10 min, and bacterial suspension OD600 = 0.6, under which the transformation rate could reach 27%. Third, based on the "soaking co-cultivation method", a new method called the "wrapping co-cultivation method" was developed, which does not require tissue cultures and can induce transgenic hairy roots of jojoba in two months. Using the "wrapping co-cultivation method", we successfully obtained transgenic hairy roots overexpressing the ScGolS1 gene, which exhibited higher tolerance to low-temperature stress. A hairy root-based genetic transformation system of jojoba will promote the functional genomics and molecular breeding of jojoba. [ABSTRACT FROM AUTHOR]

Vine-tea plant has the scientific name Ampelopsis cantoniensis (Hook. & Arn.) Planch and is a valuable herb plant in Vietnam. This study aimed to find a method to sterilize samples and nutrient medium components in each stage of shoots regeneration, rapid shoots multiplication, complete plant creation and in vitro seedling cultivation in the nursery stage of in vitro propagation techniques of vine-tea plant. Four experiments in research room and one experiment in nursery were conducted from January to December 2023 at Hue University of Agriculture and Forestry, Vietnam. Criterias of infected, alived, clean and dead samples, rate of shoot regeneration and number of shoots/samples were measured. Research results showed that the best medium to regenerate shoots from stem section carrying dormant eyes is MS + 1.5mg BAP/l + 6.5g agar/l + 30g sucrose/l. The best medium to quickly multiply shoots is MS + 1.5 mg BAP/ l + 0.2 mg IBA/l + 6.5g agar/l + 30g sucrose/l. The best complete plant-creating medium is MS + 0.5 mg α-NAA/l + 6.5g agar/l + 30g sucrose/l. In the condition that the nursery has a rain cover roof, a black net reduced the light intensity by 70% and the tissue culture nursery substrate of humus soil under the forest canopy + coconut fiber powder + smoked rice husk (1:1:1) gave the best results with survival rate reached 76.67%. [ABSTRACT FROM AUTHOR]

Background: To confirm the diagnosis of periprosthetic joint infection (PJI), the Infectious Diseases Society of America (IDSA) and the International Consensus Meeting (ICM) have defined criteria that include histology as a minor criterion and the sonication method only as an additional criterion. The aim of this monocentric, retrospective study was to investigate the value of histology and whether sonication leads to a more accurate diagnosis. Materials and methods: All revision surgeries for knee and hip arthroplasty between 2017 and 2020 were included. With regard to microbiological diagnostic, conventional culture of periprosthetic biopsies and sonication of explant material were performed. In addition, histology and non-specific inflammatory markers (CRP, leukocytes) were recorded. Results: A total of 78 patients with PJI and 62 aseptic controls were included. From both microbiological methods (conventional culture / sonication), Staphyloccus (S.) epidermidis and S. aureus were detected most frequently. However, compared to the conventional microbiology, a higher sensitivity was calculated for sonication, albeit with a lower specificity in relation to a PJI. In two logistic regression models for the significance of all diagnostic parameters in PJI, the AUC was 0.92 and 0.96 with histology in particular making the decisive contribution in both models (p < 0. 001, both models). Conclusion: Since histology showed the highest accuracy in the current study, its importance in the PJI criteria should be reevaluated. Sonication shows a high sensitivity for germ detection with a lower specificity and should only be used in combination with the conventional culture for microbiolgical diagnostics. [ABSTRACT FROM AUTHOR]

Actinomycetoma, a neglected tropical disease affecting the skin and soft tissues, is primarily caused by filamentous bacteria including Nocardia species. Here, we report a healthy 56-year-old man who has a one-year history of nodular lesions with seropurulent discharge on his right knee. Despite negative initial tissue culture, the sulfur granules that were partially acid-fast and Gram-positive branching filamentous rods were revealed in the tissue section. Repeated investigation identified the rare pathogen Nocardia wallacei, using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and 16S ribosomal ribonucleic acid (rRNA) gene sequencing. The patient was successfully treated with a six-month course of trimethoprim-sulfamethoxazole. This report describes a rare case of actinomycetoma due to N. wallacei, highlighting the challenges in diagnosis and the importance of accurate pathogen identification for the successful management of infection. The current literature regarding the causative agent will also be discussed. [ABSTRACT FROM AUTHOR]

Optimizing concentrations of minerals and phytohormones is essential when culturing a new plant species. The objective of this study was to use Response Surface Methodology (RSM) to evaluate combinations of selected minerals (N, Ca, and P) along with zeatin (Zea) to obtain optimum shoot growth of little-leaf mockorange. Forty-six treatment combinations were assigned using Proc Optex in SAS software version 9.4. The concentrations of Zea tested were 0.82, 1.095, or 1.37 µM, and the minerals were 22.5, 30, or 37.5 mM N, 1.13, 1.5, or 1.875 mM Ca, and 0.31, 0.625, or 0.937 mM P. Treatment concentrations were tested for their effects on the number of axillary shoots formed, shoot length, and dry weight. The response surface analyses showed that the optimum concentrations of N, Ca, and P were 34 to 39 mM, 1.5 mM, and 0.625 mM, respectively. Medium supplemented with 1.1 µM Zea affected shoot growth positively. Comparison of mineral concentrations in medium with concentrations in full-strength Murashige and Skoog (MS) medium, suggests ½ MS medium should be appropriate to efficiently multiply little-leaf mockorange shoots efficiently, thus saving the time and money involved in creating a custom medium formulation. [ABSTRACT FROM AUTHOR]

Cannabis sativa L. cv. 'Cheungsam' is an industrial hemp plant of Republic of Korea origin, primarily cultivated for fiber and seed production. In vitro seed germination and tissue culture are valuable tools for developing various biotechnological techniques. In the present study, we aimed to develop a tissue culture process for hemp plants using Cheungsam as a model plant and examine the secondary metabolites produced from its callus. We also developed a method to prepare pathogen-free seedlings from field-derived seeds using hydrogen peroxide (H2O2) solution as a liquid germination medium. Treating seedlings with removed seed coat in 3% H2O2 significantly reduced the contamination rate. Callus formation and de novo organogenesis of shoots and roots from callus were successfully achieved using cotyledon and leaf tissues prepared from the pathogen-free seedlings. The most effective in vitro regeneration results were obtained using the Murashige and Skoog (MS) medium supplemented with certain targeted growth regulators. An optimal combination of 0.5 mg/L thidiazuron (TDZ) and 1.0 mg/L 1-naphthalene acetic acid proved highly effective for callus induction. The addition of 0.5 mg/L TDZ in the MS medium significantly stimulated shoot proliferation, while robust root development was best supported by MS medium supplemented with 2.5 mg/L indole-3-butyric acid for both cotyledon and leaf explants. Finally, gas chromatography–mass spectrometry (GC–MS) analysis of ethanol extract from Cheungsam leaf callus revealed the presence of different secondary metabolites, including 9-octadecenamide, methyl salicylate, dodecane, tetradecane, and phenol, 2,4-bis-(1,1-dimethylethyl). This study provides a comprehensive de novo regeneration protocol for Cheungsam plants and insight into the secondary metabolite profiles of its callus. [ABSTRACT FROM AUTHOR]

Previously, a synchrotron‐based horizontal proton beamline (87.2 MeV) was successfully commissioned to deliver radiation doses in FLASH and conventional dose rate modes to small fields and volumes. In this study, we developed a strategy to increase the effective radiation field size using a custom robotic motion platform to automatically shift the positions of biological samples. The beam was first broadened with a thin tungsten scatterer and shaped by customized brass collimators for irradiating cell/organoid cultures in 96‐well plates (a 7‐mm‐diameter circle) or for irradiating mice (1‐cm2 square). Motion patterns of the robotic platform were written in G‐code, with 9‐mm spot spacing used for the 96‐well plates and 10.6‐mm spacing for the mice. The accuracy of target positioning was verified with a self‐leveling laser system. The dose delivered in the experimental conditions was validated with EBT‐XD film attached to the 96‐well plate or the back of the mouse. Our film‐measured dose profiles matched Monte Carlo calculations well (1D gamma pass rate >95% with the criteria of 2%/1 mm/2% dose threshold). The FLASH dose rates were 113.7 Gy/s for cell/organoid irradiation and 191.3 Gy/s for mouse irradiation. These promising results indicate that this robotic platform can be used to effectively increase the field size for preclinical experiments with proton FLASH. [ABSTRACT FROM AUTHOR]

Tick-borne pathogens are growing in importance for human and veterinary research worldwide. We developed, optimized, and validated a reliable quantitative PCR (qPCR; real-time PCR) assay to assess Borrelia burgdorferi infection by targeting two B. burgdorferi genes, ospA and flaB. When assessing previously tested tick samples, its performance surpassed the nested PCR in efficiency, sensitivity, and specificity. Since the detection of Borrelia is more difficult in mammalian samples, the qPCR assay was also assessed using wildlife tissues. For wildlife samples, the sensitivity and specificity of ospA primers, with the incorporation of a pre-amplification step, was equivalent or superior to the nested PCR. For human samples, no primer set was successful with human tissue without culture, but we detected Borrelia with ospA and flaB primers in 50% of the Lyme culture samples, corresponding to 60% of the participants with a Lyme disease diagnosis or suspicion. The specificity of amplification was confirmed by Sanger sequencing. The healthy participant culture samples were negative. This PCR-based direct detection assay performs well for the detection of Borrelia in different biological samples. Advancements in detection methods lead to a better surveillance of Borrelia in vectors and hosts, and, ultimately, enhance human and animal health. [ABSTRACT FROM AUTHOR]

Recent advances in tissue engineering have been remarkable, yet the precise control of cellular behavior in 2D and 3D cultures remains challenging. One approach to address this limitation is to genomically engineer optogenetic control of cellular processes into tissues using gene switches that can operate with only a few genomic copies. Here, we implement blue and red light-responsive gene switches to engineer genomically stable two- and three-dimensional mammalian tissue models. Notably, we achieve precise control of cell death and morphogen-directed patterning in 2D and 3D tissues by optogenetically regulating cell necroptosis and synthetic WNT3A signaling at high spatiotemporal resolution. This is accomplished using custom-built patterned LED systems, including digital mirrors and photomasks, as well as laser techniques. These advancements demonstrate the capability of precise spatiotemporal modulation in tissue engineering and open up new avenues for developing programmable 3D tissue and organ models, with significant implications for biomedical research and therapeutic applications. Recent advances in tissue engineering have been remarkable, yet the precise control of cellular behavior in 2D and 3D cultures remains challenging. Here by using light signals, they spatially regulated programmed cell death pathways and cell-cell communication cascades in 2- and 3-dimensional designer tissues. [ABSTRACT FROM AUTHOR]

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Introduction: Tuberculosis is prevalent in high-burden countries. However, spinal multi-drug resistant tuberculosis (MDR-TB) in patients with normal immune function is a disease that is prone to misdiagnosis and even delayed diagnosis. Recently, we successfully treated one such patient. Case Presentation: A 46-year-old male patients with lower back pain associated with recurrent fever 2 months before admission. The patient was misdiagnosed as a suppurative spinal infection and failed to respond to treatment for 2 months. The muscle strength of both lower limbs decreased progressively. We performed two operations to clear the lesion and decompress the spinal canal, during which we found a fish-like inflammatory tissue mimicking a suppurative infection. Finally, the patient was diagnosed with lumbar MDR-TB by culture, Xpert MTB/RIF and metagenomic next-generation sequencing (mNGS). The second-line anti-tuberculosis treatment (ATT) is cycloserine + para-aminosalicylic acid + ethambutol + levofloxacin + linezolid. Finally, the patient's symptoms were relieved and the muscle strength of both lower limbs recovered. Conclusion: This case prompt MDR-TB of the spine is not a typical clinical symptoms and imaging examination is the lack of specificity, when for the diagnosis of patients with spinal bone destruction unclear or treatment is invalid, can diagnostic anti-tuberculosis treatment. For patients with spinal instability or spinal canal occupying, early surgical removal of lesions, tissue culture, Xpert MTB/RIF and mNGS to identify pathogens and drug resistance, timely diagnosis and treatment can maximize the prognosis of spinal MDR-TB. [ABSTRACT FROM AUTHOR]

The current study aims to assess the callus induction and regeneration of shoots and roots in Hyssopus officinalis L. using sodium nitroprusside (SNP) and plant growth regulators (PGRs). A significant impact was found in the callogenesis of leaf and petiole explants with different concentrations of SNP and 2,4-dichlorophenoxyacetic acid (2,4-D). SNP concentrations of 20 µm induced callogenesis to 72%, but when combined with 2,4-D concentrations, callogenesis increased to 100%. In the absence of SNP, no calluses were developed. A significant effect of 2,4-D and SNP was observed on callus fresh weight. Callus fresh weight from leaf explants was 0.165 g, while from petiole explants was 0.13 g. Furthermore, SNP enhanced shoot regeneration of leaf explants compared to petiole explants. When 20 µm of SNP was combined with 3 mg/L 6-benzylaminopurine (BAP), the maximum mean shoot number was obtained (84% in leaves and 54% in petioles). A combination of SNP and IBA enhanced rooting rate to 83% and root length to 8.3 cm. After acclimation, over 80% of the produced plants survived in greenhouse conditions. Several implications can be drawn from the study's findings regarding medicinal hyssop cultivation, the preservation of genetic resources, and pharmaceutical research on medicinal plants. [ABSTRACT FROM AUTHOR]

The potential of nanotechnology revolutionized the different fields of modern sciences, enabling researchers to develop nanomaterial with extraordinary functionalities making life easier. Nanoparticles such as ZnO NPs and Ag NPs along with sterilizing agents found great potential for controlling contamination in banana tissue culture. Banana provides food for millions of people worldwide even though its origin was South East Asia and the Indian subcontinent. The emergence of tissue culture technology replaces the conventional banana propagation method, i.e., using suckers. Contaminations persist in tissue culture due to nutrient-rich media, thus decreasing its efficiency and profitability. The motive of this study is to make banana tissue culture more profitable by eradicating the contamination problem. Surface-sterilized explant may not be germ-free from inside due to covert bacteria (endogenous bacteria), which reside inside the plant in a passive state but can contaminate the experimental setup at later stages. In the present study, an experiment is designed by using various sterilizing agents (ethanol, NaOCl, H2O2, and HgCl2) for different periods to optimize the best combination of sterilizing agents for banana tissue culture. After treatment with these chemicals, the explant was inoculated to Murashige and Skoog (MS) media with plant growth hormones such as kinetin and indole acetic acid (IAA) and placed into a growth chamber for 1 week and observed regularly for appearance of contamination. The result shows that when the explant was treated with 70% ethanol for 1 min, 20% NaOCl for 10 min, and 0.2% HgCl2 for 2 min, we found it to be the most effective with the least contamination, i.e., 20%. In a separate experiment, zinc oxide (ZnO) and silver (Ag) nanoparticles were used to control endophytic contamination with different concentrations (60 mg/liter, 90 mg/liter, 120 mg/liter). Both nanoparticles show promising results in controlling endophytic contamination, but using silver nanoparticles as a part of media controls up to 100% contamination. Dual treatment of ZnO and Ag nanoparticles as sterilizing agents as an additional component of media successfully removes the contamination but becomes the major cause of tissue death in banana tissue culture. Chemical sterilization was good but not efficient as compared to the combination with nanoparticle application. Blends of chemicals and nanoparticles are effective against microbes not only from the surface but also from the inside of the plant, so using available nanoparticles with optimized concentration may open a new way in tissue culture technology. The result of this research explains the success of nanoparticles (Ag NPs and ZnO NPs) in controlling exo and endophytic contaminations in banana tissue culture, thus creating a unique way of banana explant sterilization. [ABSTRACT FROM AUTHOR]

Eucalyptus cloeziana is an important, fast-growing, precious timber species in southern China, with tissue culture being the primary method for its propagation. However, the declining proliferation coefficient of adventitious shoots with multi-generation culture is a major constraint on its rapid propagation. This study aims to address this issue through the selection of suitable explants and optimizing the plant growth regulators' formulation during the process of shoot proliferation. In this study, we cut shoots from the 21st generation of the tissue-cultured seedlings of 'Chuanlinzhen 7523' into apical, middle, and basal sections, and we measured their content of endogenous hormones. The proliferation coefficient (PC) and growth coefficient (GC) of explants under different concentrations of plant growth regulators were analyzed and ranked using the PCA method. The results indicated that the shoot basal sections were the best for proliferation when treated with 0.40 mg/L BAP (6-benzylaminopurine), 0.20 mg/L NAA (naphthaleneacetic acid), and 0.20 mg/L TDZ (thidiazuron), yielding a PC of 4.7 and a GC of 4.1. These basal sections ranked first with a rooting rate of up to 80%. When the rooted plantlets were acclimatized in the nursery, a survival rate of 100% was achieved. This protocol—from proliferation to acclimation—effectively improves the propagation efficiency of E. cloeziana 'Chuanlinzhen 7523' after multi-generation propagation. [ABSTRACT FROM AUTHOR]

Background: Early confirmation of infections with influenza virus and/or respiratory syncytial virus (RSV) is beneficial for prompt treatment and outbreak management. This study aimed to assess the Cepheid Xpert Xpress Flu/ RSV assay in Central China, using Sanger sequencing as the reference method. Methods: Nasopharyngeal swab (NP) samples from pediatric and adult patients with influenza-like illnesses were collected by the Hubei Province Disease Control and Prevention Center. The Xpert Xpress Flu/RSV assay was performed according to the manufacturer's guidelines, and the cycle threshold (Ct) values of each positive sample were recorded. Sanger sequencing was then performed on the NP samples, and discordant results were verified with a self-built fluorescent PCR method and virus tissue culture. Results: The Xpert assay demonstrated a sensitivity of 97.06%, a specificity of 95.81%, an accuracy of 95.93%, a PPV of 71.74%, and a negative predictive value (NPV) of 99.66% for influenza A (Flu A) detection. Influenza B (Flu B) detection had a sensitivity of 100%, a specificity of 88.52%, an accuracy of 91.86%, a PPV of 78.12%, and an NPV of 100%. RSV detection had a sensitivity of 100%, a specificity of 95.67%, an accuracy of 98.80%, a PPV of 60.00%, and an NPV of 100%. False positive samples had significantly higher mean Ct values compared to true positive samples for detecting Flu A, Flu B, or RSV (p < 0.001). Conclusions: The Xpert Xpress Flu/RSV test is a rapid, accurate, and reliable molecular diagnostic method for Flu A, Flu B, and RSV. Positive Xpert results with a low Ct value are more likely to indicate a clinical infection with Flu/RSV to allow timely diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Biofilm is a structurally‐connected microbial community, covered by a self‐produced polymeric matrix and adhered to biotic or abiotic surfaces. This study aimed to evaluate the occurrence of biofilm‐producing antibiotic‐resistant bacteria in water from distribution systems. Water samples were taken from 32 tanks across Ondo City and Akure metropolis, Nigeria. Information regarding the sanitation status of the tanks was gathered by observation and oral interviews. The physicochemical properties were determined using standard methods. Using the pour plate technique. Agars included serially diluted water samples were inoculated onto plate count agar, mannitol salt agar, Salmonella-Shigella agar, MacConkey agar, and cetrimide nutrient agar to assess total viable bacteria, Staphylococcus aureus, Salmonella and Shigella, coliforms, and Pseudomonas aeruginosa, respectively. Eosin‐methylene blue agar was used to cultivate Escherichia coli and Enterobacter aerogenes. Pure isolates were characterised using API kits and assessed for antibiotic resistance and biofilm production employing the Kirby‐Bauer and tissue culture plate techniques, respectively. The ages of the water tanks ranged from 1 to 25 years old; all tanks had cover‐lids; 13 (40.63%) had water guards while 12 (37.5%) underwent water treatment. The physicochemical properties chiefly fell within WHO standards for drinking water. One hundred and eighty‐seven isolates were obtained. S. aureus (15.51%) had the highest frequency while Salmonella enterica (3.2%) had the lowest frequency. Thirty‐six percent of the isolates were strong biofilm producers, while 20.67% Gram‐negative and 18.69% Gram‐positive bacterial isolates were antibiotic‐resistant. This study revealed a high occurrence of biofilm‐forming bacteria and prevalence of antibiotic‐resistant bacteria in water distribution systems, emphasizing the urgency of improving water quality for public health protection. [ABSTRACT FROM AUTHOR]

Introduction: Porcine circovirus type 2 (PCV2) is an important pathogen that causes diarrhea in nursery and fattening pigs, resulting in huge economic losses for commercial pig farms. Protective efficacy of vaccines is compromised by mutations in pathogens. There is an urgent need to articulate the mechanism by which PCV2 destroys the host's intestinal mucosal barrier and to find effective therapeutic drugs. Increasing attention has been paid to the natural antiviral compounds extracted from traditional Chinese medicines. In the present study, we investigated the role of Matrine in mitigating PCV2-induced intestinal damage and enhancing autophagy as a potential therapeutic strategy in mice. Methods: A total of 40 female, specific-pathogen-free-grade Kunming mice were randomly divided into four groups with 10 mice in each group: control, PCV2 infection, Matrine treatment (40 mg/kg Matrine), and Ribavirin treatment (40 mg/kg Ribavirin). Except for the control group, all mice were injected intraperitoneally with 0.5 mL 105.4 50% tissue culture infectious dose (TCID50)/mL PCV2. Results: While attenuating PCV2-induced downregulation of ZO-1 and occludin and restoring intestinal barrier function in a PCV2 Kunming mouse model, treatment with Matrine (40 mg/kg) attenuated ultrastructural damage and improved intestinal morphology. Mechanistically, Matrine reversed PCV2-induced autophagosome accumulation by inhibiting signal transducer and activator of transcription 3 (STAT3) phosphorylation and upregulating Beclin1 protein expression, thus resisting viral hijacking of enterocyte autophagy. Discussion: Our findings demonstrate that Matrine may be a novel, potential antiviral agent against PCV2 by activating intestine cellular autophagy, which provides a new strategy for host-directed drug discovery. [ABSTRACT FROM AUTHOR]

Introduction: The endometrium plays a crucial role in early human pregnancy, particularly in embryo implantation, survival, and growth. However, invasion and infection by pathogens can lead to endometritis, infertility, and poor reproductive outcomes. Understanding the mechanisms of endometritis and its impact on fertility remains limited. An infection model using patient-derived endometrial epithelial gland organoids (EEGOs) was established to advance in vitro studies on endometritis and related infertility. Methods: An EEGOs infection model was constructed and characterized from human endometrium, treating the organoids with estrogen and progesterone to observe changes in the proliferative and secretory phases. The organoids were infected with E. coli , and the release of inflammatory cytokines in the supernatant was detected using ELISA. RNA-seq was employed to analyze the differences before and after E. coli treatment, and differential gene mRNA expression was validated using real-time quantitative PCR. Additionally, the effect of E2 in alleviating inflammation was assessed through markers of receptivity (PAEP, LIF, ITGβ), proliferation (Ki67), and barrier repair (ZO-1). Results: The constructed human EEGOs exhibited long-term expansion capability, genetic stability, and characteristic hormonal responses, strongly expressing epithelial markers (MUC1, E-Cadherin). After E. coli infection, the expression levels of inflammatory cytokines TNF-α, IL-8, and IFN-γ increased significantly (P < 0.05). RNA-seq indicated that the MAPK signaling pathway was activated post-infection, with increased expression levels of heat shock proteins and transcription factor mRNA. E2 treatment post-infection significantly decreased the mRNA expression of inflammatory genes IL-1β, IL8, IL6 and TNF-α compared to the E. coli infected group (P < 0.05). Additionally, the expression of genes related to receptivity, proliferation, and barrier repair was enhanced in the E2-treated organoids. Conclusions: Our findings demonstrate that patient-derived EEGOs are responsive to bacterial infection and are effective models for studying host-pathogen interactions in bacterial infections. These organoids revealed the anti-inflammatory potential of E2 in alleviating E. coli -induced inflammation, providing insights into the mechanisms of endometritis and its impact on infertility. The study supports the use of EEGOs as valuable tools for understanding endometrial health and developing targeted treatments. [ABSTRACT FROM AUTHOR]

The seedlings of mature Ribes maximowiczianum Komarov seed germination under different disinfection treatment were used as explants to study the influence of different plant growth regulators on buds induction, hyperplasia, sound seedling and rhizogenesis. The results showed that 500 mg/L GA3 soaking treatment was conducive to seed germination, and the germination rate could reach 66.7%, and the best disinfection treatment was 75% alcohol for 30s and 0.1% mercuric acid disinfection for 8 min, the contamination rate was 23.3%. The best medium for bud induction was MS + 2.0 mg/L 6-BA+ 0.05 mg/L NAA, the proliferation times was 7.5. The best medium for rooting was 1/2MS+2.0 mg/L IBA, the rooting rate was 100%, and the average number of rooting shoots was 6.33. The survival rate was 87.5% in the medium of 3:1:1 in peat soil: coir: perlite. This study established a complete system of tissue culture and rapid propagation for Ribes maximowiczianum Komarov seed, which is suitable for the large-scale production and application of Ribes maximowiczianum Komarov seedlings. [ABSTRACT FROM AUTHOR]

This study used the leaves, stem segments, and bracts of alpine rhododendron varieties ‘23XXL’‘5RE’and‘6GR’ as explants to investigate the effects of basic culture medium, physiological age of explants, and plant growth regulators on callus induction in alpine rhododendron explants. The results showed that the explants of alpine rhododendron could induce callus on both WPM and MS media. During the callus induction stage, adding 2.0-3.0 mg/L TDZ had a good effect on inducing callus. Among them, the callus induced by leaves with a physiological age of 2W had better growth, and the highest callus rate was 100%;at the stage of callus differentiation, the most suitable ratio of callus differentiation hormones was ZT 2.5 mg/L+NAA 0.5 mg/L and TDZ 1.5 mg/L+NAA 1.0 mg/L. Among them, the explant with a physiological age of 1M had better callus differentiation on Z7 medium, with an average number of bud differentiation of 0.92;during the rooting stage, the optimal hormone ratio of the culture medium was IBA 0.5 mg/L+NAA 1.0-1.5 mg/L, which were suitable for the three varieties of alpine rhododendron. The research results provide a scientific basis for constructing an efficient and stable tissue culture and rapid propagation system of alpine rhododendron, and lay a foundation for further realizing the industrialization of alpine rhododendron. [ABSTRACT FROM AUTHOR]

Obesity, a prevalent global health issue, arises from an imbalance between caloric intake and energy expenditure, leading to the expansion of adipose tissue and metabolic dysfunction. White adipose tissue (WAT) stores energy as lipids, while brown adipose tissue (BAT) plays a pivotal role in energy dissipation through adaptive thermogenesis. Recent research initiatives have focused on finding strategies to decrease adipogenesis and fat mass accumulation and increase thermogenesis. Finding chemicals with anti-obesity properties would be beneficial. Resveratrol, a polyphenolic compound abundantly found in the skin of grapes and red wine, possesses anti-oxidant, anti-inflammatory, anti-cancer, and anti-obesity properties. This literature review examines the effects of resveratrol on adipocytes in culture and adipose tissue in animal models of obesity. The existing evidence indicates that resveratrol may exert its anti-obesity effects by inhibiting adipogenesis, promoting the apoptosis of mature adipocytes, reducing lipid accumulation, and increasing thermogenesis. Further research utilizing animal and clinical studies is required to understand in detail the anti-obesity potential of resveratrol. [ABSTRACT FROM AUTHOR]

Simple Summary: Glioblastoma is a highly aggressive brain cancer with limited treatment options. This research aims to improve our understanding of GBM progression by focusing on extracellular vesicles released by tumor cells that carry important biological information. The authors developed a laboratory model that closely mimics the tumor microenvironment, allowing for the study of extracellular vesicles released from various cell types within the tumor. This model enabled the identification of specific surface markers on extracellular vesicles that could serve as potential noninvasive diagnostic tools for GBM. The findings suggest that these markers may help improve extracellular vesicle isolation and provide insights into the molecular characteristics of the tumor. This study has the potential to significantly impact the research community by offering new approaches to GBM diagnosis and advancing our knowledge of tumor biology, which could ultimately inform future therapeutic strategies. Background/Objectives: Glioblastoma (GBM) is an aggressive brain cancer with limited treatment options. Extracellular vesicles (EVs) derived from GBM cells contain important biomarkers, such as microRNAs, proteins, and DNA mutations, which are involved in tumor progression, invasion, and resistance to treatment. Identifying surface markers on these EVs is crucial for their isolation and potential use in noninvasive diagnosis. This study aimed to use tumor-derived explants to investigate the surface markers of EVs and explore their role as diagnostic biomarkers for GBM. Methods: Tumor explants from nine GBM patients without IDH1/IDH2 mutations or 1p-19q co-deletion were cultured to preserve both tumor viability and cytoarchitecture. EVs were collected from the tumor microenvironment using differential centrifugation, filtration, and membrane affinity binding. Their surface protein composition was analyzed through multiplex protein assays. RNA-Seq data from TCGA and GTEx datasets, along with in silico single-cell RNA-seq data, were used to assess EV surface biomarker expression across large GBM patient cohorts. Results: The in vitro model successfully replicated the tumor microenvironment and produced EVs with distinct surface markers. Biomarker analysis in large datasets revealed specific expression patterns unique to GBM patients compared with healthy controls. These markers demonstrated potential as a GBM-specific signature and were correlated with clinical data. Furthermore, in silico single-cell RNA-seq provided detailed insights into biomarker distribution across different cell types within the tumor. Conclusions: This study underscores the efficacy of the tumor-derived explant model and its potential to advance the understanding of GBM biology and EV production. A key innovation is the isolation of EVs from a model that faithfully mimics the tumor's original cytoarchitecture, offering a deeper understanding of the cells involved in EV release. The identified EV surface markers represent promising targets for enhancing EV isolation and optimizing their use as diagnostic tools. Moreover, further investigation into their molecular cargo may provide crucial insights into tumor characteristics and evolution. [ABSTRACT FROM AUTHOR]

Background: Peucedanum praeruptorum Dunn has typical stacked umbels and medicinal value; however, the lack of an effective tissue culture system for P. praeruptorum has limited the large-scale propagation of its seedlings. Results: We systematically established an in vitro regeneration system for P. praeruptorum using young leaves and stems as explants. Tissue culture plantlets were successfully obtained within 123 and 90 d of somatic embryogenesis and organogenesis, respectively. Combined plant growth regulators (PGRs) were optimized to promote efficient plant regeneration at each stage of the culture process. Specifically, embryogenic callus induction was superior in Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 6-benzyladenine (BA) and 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). For somatic embryonic development, the highest differentiation rates were achieved using BA, 2,4-D, and 6-furfuryl aminopurine (6-KT). Induction of organogenesis resulted in the highest differentiation rates and proliferation coefficients of buds in MS medium supplemented with BA and α-naphthaleneacetic acid (NAA). Moreover, regeneration of P. praeruptorum seedlings was achieved by adjusting the BA and indole-3-butyric acid (IBA) concentrations in 1/2 MS medium. Conclusion: Our results provide a technical system for the rapid propagation of P. praeruptorum, which can facilitate germplasm improvement, resource conservation, and further genetic transformation of Peucedanum species. [ABSTRACT FROM AUTHOR]

Pseudorabies virus (PRV) is the pathogenic agent of pseudorabies, causing serious reproductive failure in swine. However, it is still unknown whether PRV uterine inoculation impairs blastocyst implantation. In the present study, a PRV infection mouse model was developed. Pregnant mice were inoculated with either 104 or 105 50% tissue culture infectious dose (TCID50) units of PRV on gestation day 2 (GD2). Viral DNA was detected in tissues by PCR and/or in situ hybridization. Histopathological change and expression of proinflammatory cytokines in uterus were analyzed by H.E. staining and qPCR, respectively. Apoptosis was also investigated by TUNEL assay, and the expression of apoptosis-related proteins including Bax and Bcl-2 was detected by Western blot. The results showed that intrauterine exposure of PRV on GD2 reduced the number of embryo implantation site. Abundant viral DNA was detected in spinal marrow and brain, and small amounts of PRV genomes were detected in embryo implantation site, ovary as well as thymus. Considerable inflammatory cells infiltrating in the endometrium, with high levels of pro-inflammatory cytokines of interleukin (IL)-6, IL-1β and tumor necrosis factor- α mRNA after infection. In addition, PRV infection promoted apoptosis in stroma and endothelium of the mouse endometrium. Collectively, intrauterine inoculation of PRV during early pregnancy causes cytokine release syndrome and apoptosis in endometrium, which impairs mouse embryo implantation. [ABSTRACT FROM AUTHOR]

Tumor organoids, an in-vitro three-dimensional model, possess high potential for investigating tumor biology and treatment response and have been demonstrated more appropriate for drug assessment than two-dimensional cultures. Herein, we described two cases of invasive high-grade urothelial carcinoma who underwent radical cystectomy successfully following use of patient-derived organoids (PDOs) for drug screening to inform therapeutic decisions. In these two cases, the PDOs cultured by biopsy tissues were both sensitive to the combination of gemcitabine and cisplatin. After neoadjuvant chemotherapy (NAC) with gemcitabine and cisplatin, the patients responded well, and radical cystectomy was performed successfully. No recurrence or metastasis was observed within 6 months after surgery. This small case series suggests that the patient-derived urothelial carcinoma organoids contribute to optimizing NAC options to guide personalized treatment and improve the survival outcomes. [ABSTRACT FROM AUTHOR]

The aseptic technique is essential for minimizing contamination in cell and tissue culture. However, biomedical science undergraduates in Universiti Putra Malaysia face challenges in acquiring hands-on learning experience on proper aseptic techniques due to inadequate facilities, which decreases learning interest and efficiency. To address this, an in-house virtual reality (VR) mobile application, AsepticTech VR, was developed to simulate a cell and tissue culture facility for practicing aseptic techniques. This study aims to investigate the efficacy of AsepticTech VR as a teaching and learning tool. Forty-one biomedical undergraduates were randomly assigned into test and control groups. Both groups attended a pre-test to evaluate the pre-intervention learning performances. Then, only test group participants were given access to AsepticTech VR, while both groups attended traditional lectures and practical sessions for a week. After one week, all participants took a post-test to evaluate their post-intervention learning performances. Additionally, test group participants also completed the modified Differential Emotions Scale (mDES) questionnaire to assess their emotions associated with using AsepticTech VR, and Web-Based Learning Tool (WBLT) questionnaire to assess their learning experience. The results showed that the test group participants had better learning performance improvements in the cognitive (p < 0.05) and psychomotor domain (p < 0.01) compared to the control group but did not differ in their affective domain. Moreover, the test group demonstrated stronger positive emotions compared to negative emotions (p < 0.001) and had positive learning experience with AsepticTech VR. This indicates that AsepticTech VR is an effective teaching and learning tool for cell and tissue culture aseptic techniques. [ABSTRACT FROM AUTHOR]

Paeonia ostii, a plant of substantial economic significance, continues to face constraints in achieving large-scale propagation. In vitro propagation offers a promising avenue for the production of disease-free plants and the genetic transformation of peonies to instill novel traits. However, significant challenges persist in tissue culture, particularly with regards to the reproduction coefficient of shoots and the rooting process. This study reports an efficacious protocol for P. ostii micropropagation, focusing on in vitro root development facilitated through the application of phloroglucinol (PG). Furthermore, the study unveils the molecular signature of P. ostii during in vitro root development. The results indicate that the modified Y3 medium (Y3M), supplemented with 1 mg/L 6-benzyladenine (BA) and 0.1 mg/L α-naphthaleneacetic acid (NAA), is optimal for adventitious bud induction, achieving a 96.67% induction rate and an average of 16.03 adventitious shoots per sample. The highest elongation percentage (92.15%) and the longest average shoot length (3.87 cm) were obtained with Y3M containing 0.3 mg/L BA and 0.03 mg/L NAA. Additionally, the optimal medium for inducing root formation in P. ostii was identified as WPM supplemented with 3 mg/L indole-3-butyric acid (IBA) and 100 mg/L phloroglucinol (PG). Lignin content detection, microscope inspection, and molecular signature results demonstrated that PG enhanced lignin biosynthesis, thereby promoting in vitro rooting of P. ostii. [ABSTRACT FROM AUTHOR]

Somatic embryogenesis (SE) provides alternative methodologies for the propagation of grapevine (Vitis spp.) cultivars, conservation of their germplasm resources, and crop improvement. In this review, the current state of knowledge regarding grapevine SE as applied to these technologies is presented, with a focus on the benefits, challenges, and limitations of this method. The paper provides a comprehensive overview of the different steps involved in the grapevine SE process, including callus induction, maintenance of embryogenic cultures, and the production of plantlets. Additionally, the review explores the development of high-health plant material through SE; the molecular and biochemical mechanisms underlying SE, including the regulation of gene expression, hormone signaling pathways, and metabolic pathways; as well as its use in crop improvement programs. The review concludes by highlighting the future directions for grapevine SE research, including the development of new and improved protocols, the integration of SE with other plant tissue culture techniques, and the application of SE for the production of elite grapevine cultivars, for the conservation of endangered grapevine species as well as for cultivars with unique traits that are valuable for breeding programs. [ABSTRACT FROM AUTHOR]

Degrons are minimal protein features that are sufficient to target proteins for degradation. In most cases, degrons allow recognition by components of the cytosolic ubiquitin proteasome system. Currently, all of the identified degrons only function within the cytosol. Using Saccharomyces cerevisiae, we identified the first short linear sequences that function as degrons from the endoplasmic reticulum (ER) lumen. We show that when these degrons are transferred to proteins, they facilitate proteasomal degradation through the endoplasmic reticulum associated degradation (ERAD) system. These degrons enable degradation of both luminal and integral membrane ER proteins, expanding the types of proteins that can be targeted for degradation in budding yeast and mammalian tissue culture. This discovery provides a framework to target proteins for degradation from the previously unreachable ER lumen and builds toward therapeutic approaches that exploit the highly conserved ERAD system. [ABSTRACT FROM AUTHOR]

Background : Hemsleya Chinensis is a perennial plant in the Cucurbitaceae family containing antibacterial and anti-inflammatory compounds. The lack of genetic transformation systems makes it difficult to verify the functions of genes controlling important traits and conduct molecular breeding in H. chinensis. Results: Highly efficient calli were induced on MS medium added 1.5 mg·L− 1 6-benzylaminopurine (6-BA) and 0.02 mg·L− 1 1-naphthylacetic acid (NAA) with high efficiency (> 95%). The frequency of shoot induction was increased to 90% with a plant growth regulator combination of 1.5 mg·L− 1 6-BA and 0.1 mg·L− 1 NAA. Our results also showed that 100% of shoot regeneration was achieved in a shoot regeneration medium. Additionally, more than 92% of kanamycin-resistant plants were confirmed. Furthermore, we achieved 42% genome editing efficiency by applying this transformation method to HcOSC6, a gene that catalyzes the formation of cucurbitadienol. HPLC analysis showed OE-HcOSC6 lines exhibited significantly higher cucurbitadienol levels than the genome-edited lines. Transcriptomic analysis revealed that some downstream genes related to cucurbitadienol biosynthesis, such as HcCYP87D20, HcCYP81Q58, and HcSDR34, were up-regulated in OE lines and down-regulated in mutants. Conclusions: Here, we established a process for regeneration, transformation, and genome editing of H. chinensis using stem segments. This provides valuable insight into the underlying molecular mechanisms of medicinal compound production. By combining high-efficiency tissue culture, transformation, and genome editing systems, we provide a powerful platform that supports functional research on molecular mechanisms of secondary metabolism. [ABSTRACT FROM AUTHOR]