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Background: DiGeorge syndrome (DGS) is a condition typically associated with athymia, parathyroid hypoplasia or aplasia, and congenital heart defects. Athymia in these patients causes severe immunodeficiency, causing high mortality and morbidity, often requiring thymic tissue transplantation. The present systematic review aims to consolidate the present evidence on thymus transplantation in DGS. Methods: An electronic literature search of five databases (PubMed, Medline, Scopus, EBSCOhost, and CINAHL) was performed from inception till September 2024. Relevant articles were selected, and data was extracted by two independent reviewers. Results: A total of 16 articles were included from an initial set of 1227 articles. Patients diagnosed with DGS in the included studies were predominantly male, and the age at which thymus transplantation was done typically varied from 0.8 to 26 months. Several patients had chromosome 22q11 hemizygosity. Thymic tissue was taken from tissues of pediatric patients undergoing cardiothoracic surgery. Pre-transplant medication included immunosuppressants with rabbit anti-thymocyte globulin (RATGAM) being frequently used alongside steroids and tacrolimus. This tissue was cultured and transplanted into the quadriceps muscle of the patients under general anesthesia. Thymopoiesis was well described in most patients with graft failures and rejections occurring rarely. Naive T-cell development was noted in almost all patients with clearance of infections in many cases. Common postoperative complications include sepsis, haemorrhage, gastrointestinal disturbances, among others. Mortality was uncommon but often associated with intracerebral hemorrhages and sepsis. Conclusion: Thymus transplantation is a relatively safe and effective procedure in patients with DGS with athymia. Future research should explore the addition of allogenic parathyroid gland transplantation along with thymic tissue. [ABSTRACT FROM AUTHOR]

Polyploidy, characterized by chromosome multiplication, plays a pivotal role in plant evolution and agricultural innovation. This review explores advanced methodologies for tetraploid development in tissue culture, highlighting chemical inducers such as colchicine and oryzalin, physical strategies including thermal and radiation treatments, and cutting-edge approaches like CRISPR-Cas9. These techniques facilitate targeted genetic modification, enhancing traits such as stress tolerance, disease resistance, and biomass in critical crops like garlic, wheat, and ornamentals. Despite substantial advancements, challenges including chromosomal instability and growth abnormalities necessitate protocols. This review emphasizes polyploidy’s transformative potential for agricultural productivity, aligning with global food security imperatives while advocating for ongoing innovation and optimization. [ABSTRACT FROM AUTHOR]

Key message: In the MS medium containing flax plant extract, the endogenous GA and IAA hormone content in the explant tissues is stimulated and a high regeneration rate and rooting are achieved. In vitro regeneration efficiency is directly related to the endogenous hormone level of the plant. In this context, the potential regeneration efficiency of explants can be maximized by utilizing the endogenous plant hormone content in in vitro tissue culture studies. In this study, the effects of plant extracts obtained from flax (Linum usitatissimum L.) varieties were investigated on the in vitro regeneration capacity, rooting performance and tissue hormone content of flax cultivars. The Clark, Madaras and 1886 Sel. varieties were used as explants and plant extract sources. Abscisic acid (ABA), gibberellic acid (GA) and indole-3-acetic acid (IAA) contents of plant extracts were analyzed by the HPLC method. In vitro regeneration experiments were carried out in Murashige and Skoog (MS) media containing four different plant extract doses (0, 5 ml l−1, 10 ml l−1 and 15 ml l−1) and chemical control. Tissue endogenous GA and IAA contents were determined in two stages as shoot regeneration (SH) and rooting stage (RH). Differences were observed in the examined regeneration parameters and endogenous hormone content among the varieties. The best shoot regeneration rate in all varieties was obtained from the medium containing 10 ml l−1 plant extract. Besides, the best rooting rate for all varieties was determined in the medium containing 15 ml l−1 plant extract. The high GA content of SH tissues decreased in the RH stage. On the other hand, the tissue IAA content was high in the RH stage, and the GA content decreased or remained constant. The effect of the plant extract on the endogenous hormone content was higher than the chemical control. The results obtained proved that the phytohormone content of the plant extract increased regeneration success by stimulating the endogenous hormone content in the tissues. Determining the effect of plant extracts on tissue regeneration will contribute to evaluating these components as bio-fertilizers or bio-stimulants. [ABSTRACT FROM AUTHOR]

Nervous necrosis virus (NNV) of the genus Betanodavirus is one of the simplest RNA viruses pathogenic to a wide range of fish species. We established the SeGF, SeGE-22 and SeGB cell lines persistently infected with NNV (PI-SeGFNNV, PI-SeGE-22NNV and PI-SeGBNNV cells) by repeatedly subculturing the cells that survived NNV infection. The PI-SeGFNNV and PI-SeGE-22NNV cells continued to stably yield NNV in culture fluids at 106 to 107 median tissue culture infectious dose (TCID50)/ml even after 30–50 subcultures. The PI-SeGBNNV cells initially yielded NNV at 103.9 TCID50/ml but stopped yielding NNV after several passages. No significant morphological differences were observed between the naïve and PI-cells in either cell line. Antiviral activity suppressing the multiplication of NNV was detected in the culture fluids of all PI-cell lines. It significantly suppressed the growth (metabolism) of each cell line but did not directly influence NNV infectivity. However, this antiviral substance was not an interferon but a heat-stable (100 ºC for 3 min), small molecule with Mr < 1000. When the PI-SeGBNNV cells stopped yielding NNV after subculturing several times, the production of the antiviral substance also ceased, indicating that the production of antiviral substance is initiated by NNV infection. [ABSTRACT FROM AUTHOR]

Infantile hemangiomas (IH) are a common entity encountered by dermatologists, otolaryngologists, and other surgeons. Oral propranolol is a mainstay of treatment for IH and is well-tolerated, though propranolol-refractory IH and other drug-related adverse events are documented and can limit its usage. There are few in vitro testing systems for putative treatment agents. To address this, we modified a tissue culture system for human hemangioma treatment testing to evaluate the treatment impact of the immune modifier, imiquimod. Human umbilical vein endothelial cells (HUVEC) and hemangioma cultures were treated with several concentrations of imiquimod followed by MTT assays, reporter gene assays, PCR, ELISA, and Western blotting for IL-8, VEGF, Cyclin D1, and IFNα and immunohistochemistry for Cyclin D1 and Ki-67. HUVEC showed acute decreases in IL-8, VEGF, and Cyclin D1 promoter activity and increases in IFNα mRNA after imiquimod treatment. Hemangioma samples showed no change in Ki-67 or Cyclin D1 staining after treatment with imiquimod after 27 d, with significantly increased IL-8 and VEGF. From this preliminary analysis, we discerned that hemangioma tissues can be grown in tissue culture and used for drug treatment studies. We also conclude acute and chronic modulation of cell cycle, angiogenesis factors, and immunostimulatory conditions may be associated with imiquimod mechanisms of action in hemangioma involution. [ABSTRACT FROM AUTHOR]

Cryopreservation enhances the availability of "off-the-shelf" cell therapies. However, the choice between tissue culture polystyrene (TCP) and hollow fiber system (HFB) system for adipose-derived stem cell (ASC) production remains a critical decision, with implications for scalability, reproducibility, and the clinical efficacy. Therefore, the characteristics of ASCs expanded in TCP and HFB and cryopreserved were compared. TCP and HFB cultures were established, and cells were cryopreserved. Surface markers were analyzed to identify immunophenotypic changes and subpopulations. Clonogenicity, differentiation capability, and proliferation potentials were determined along with surrogate tests on wound healing. The expressions of the most markers were consistent before and after thawing for both systems. However, CD105 expression of TCP cells was significantly decreased by the freeze-thawing procedure. Also, CD274 was significantly less expressed on HFB-expanded cells before freezing, however, post-thawing, the proportion of CD274 positive cells was comparable to TCP cells. Besides, two expansions supported different subpopulations, influencing the heterogeneity within ASC cultures. Despite this heterogeneity, no statistical differences were observed as for ASC functional characteristics and the effects on fibroblasts. This study highlighted freeze-thaw does not interfere with the production of fully functional ASCs in either system, although it drives some differential changes in the subpopulations between systems. [ABSTRACT FROM AUTHOR]

Purpose: Necrotizing otitis externa (NOE) is a serious life-threatening infection, with Pseudomonas (PA) aeruginosa being the primary causative agent. Over the last two decades the use of systemic anti-PA antibiotics expanded substantially and are now prescribed regularly by physicians. Meanwhile, studies indicate shifting trends in the incidence of the offending pathogen in NOE. The objective of the study is to assess whether preadmission antibiotic treatment influence the incidence offending pathogens and sterile cultures in NOE. Also, we aim to evaluate the effects of preadmission antibiotic treatment on NOE severity and disease progression.A retrospective case series analysis was performed, including all patients admitted in a single tertiary center due to NOE.83 patients were included in the study. Among them 17 patients received no oral antibiotics prior to admission (NoAb-NOE), and 65 patients received systemic antibiotics prior to admission. There was no statistical difference between the groups regarding the incidence of the pathogen or sterile culture. Furthermore, there was no statistical difference regarding duration of hospitalization, need for surgery or readmission.Preadmission antibiotic treatment does not influence the incidence of offending pathogens or rate of sterile culture in NOE. Moreso, preadmission antibiotic has no effect on disease progression or severity in NOE. Our findings suggest that sterile culture NOE is probably due to the low sensitivity of superficial swabs and not secondary to partially treated disease. Deep tissue culture should be considered in the setting of NOE patients.Methods: Necrotizing otitis externa (NOE) is a serious life-threatening infection, with Pseudomonas (PA) aeruginosa being the primary causative agent. Over the last two decades the use of systemic anti-PA antibiotics expanded substantially and are now prescribed regularly by physicians. Meanwhile, studies indicate shifting trends in the incidence of the offending pathogen in NOE. The objective of the study is to assess whether preadmission antibiotic treatment influence the incidence offending pathogens and sterile cultures in NOE. Also, we aim to evaluate the effects of preadmission antibiotic treatment on NOE severity and disease progression.A retrospective case series analysis was performed, including all patients admitted in a single tertiary center due to NOE.83 patients were included in the study. Among them 17 patients received no oral antibiotics prior to admission (NoAb-NOE), and 65 patients received systemic antibiotics prior to admission. There was no statistical difference between the groups regarding the incidence of the pathogen or sterile culture. Furthermore, there was no statistical difference regarding duration of hospitalization, need for surgery or readmission.Preadmission antibiotic treatment does not influence the incidence of offending pathogens or rate of sterile culture in NOE. Moreso, preadmission antibiotic has no effect on disease progression or severity in NOE. Our findings suggest that sterile culture NOE is probably due to the low sensitivity of superficial swabs and not secondary to partially treated disease. Deep tissue culture should be considered in the setting of NOE patients.Results: Necrotizing otitis externa (NOE) is a serious life-threatening infection, with Pseudomonas (PA) aeruginosa being the primary causative agent. Over the last two decades the use of systemic anti-PA antibiotics expanded substantially and are now prescribed regularly by physicians. Meanwhile, studies indicate shifting trends in the incidence of the offending pathogen in NOE. The objective of the study is to assess whether preadmission antibiotic treatment influence the incidence offending pathogens and sterile cultures in NOE. Also, we aim to evaluate the effects of preadmission antibiotic treatment on NOE severity and disease progression.A retrospective case series analysis was performed, including all patients admitted in a single tertiary center due to NOE.83 patients were included in the study. Among them 17 patients received no oral antibiotics prior to admission (NoAb-NOE), and 65 patients received systemic antibiotics prior to admission. There was no statistical difference between the groups regarding the incidence of the pathogen or sterile culture. Furthermore, there was no statistical difference regarding duration of hospitalization, need for surgery or readmission.Preadmission antibiotic treatment does not influence the incidence of offending pathogens or rate of sterile culture in NOE. Moreso, preadmission antibiotic has no effect on disease progression or severity in NOE. Our findings suggest that sterile culture NOE is probably due to the low sensitivity of superficial swabs and not secondary to partially treated disease. Deep tissue culture should be considered in the setting of NOE patients.Conclusion: Necrotizing otitis externa (NOE) is a serious life-threatening infection, with Pseudomonas (PA) aeruginosa being the primary causative agent. Over the last two decades the use of systemic anti-PA antibiotics expanded substantially and are now prescribed regularly by physicians. Meanwhile, studies indicate shifting trends in the incidence of the offending pathogen in NOE. The objective of the study is to assess whether preadmission antibiotic treatment influence the incidence offending pathogens and sterile cultures in NOE. Also, we aim to evaluate the effects of preadmission antibiotic treatment on NOE severity and disease progression.A retrospective case series analysis was performed, including all patients admitted in a single tertiary center due to NOE.83 patients were included in the study. Among them 17 patients received no oral antibiotics prior to admission (NoAb-NOE), and 65 patients received systemic antibiotics prior to admission. There was no statistical difference between the groups regarding the incidence of the pathogen or sterile culture. Furthermore, there was no statistical difference regarding duration of hospitalization, need for surgery or readmission.Preadmission antibiotic treatment does not influence the incidence of offending pathogens or rate of sterile culture in NOE. Moreso, preadmission antibiotic has no effect on disease progression or severity in NOE. Our findings suggest that sterile culture NOE is probably due to the low sensitivity of superficial swabs and not secondary to partially treated disease. Deep tissue culture should be considered in the setting of NOE patients. [ABSTRACT FROM AUTHOR]

The efficacy of anti-cancer therapies depends on the genomic composition of the tumor, its microenvironment, spatial organization, and intra-tumor heterogeneity. B-cell lymphomas are a heterogeneous group of tumors emerging from B-cells at different stages of differentiation and exhibiting tumor-specific interactions with the tumor microenvironment. Thus, the effect of drug treatments can be influenced by the tumor composition and functional interactions among immune cells. Here, we develop a platform to maintain small fragments of human lymphoma tissue in culture for several days, and use them to test response to small molecules. We collect 27 patient samples representative of different lymphoma subtypes, and establish ex vivo tissue fragments that retain histological, cellular, and molecular characteristics of the original tissue, here referred to as lymphomoids. Using lymphomoids, we test sensitivity to several clinically approved drugs in parallel and examine tissue remodeling upon treatment. Moreover, when this information is available, we show that the effect of the inhibitors observed in lymphomoids is consistent with the patients' response in the clinic. Thus, lymphomoids represent an innovative ex vivo model to assess the effect of anti-cancer therapies while preserving the tissue structure and its components. Choosing the most effective anti-cancer therapy is difficult due to the high degree of tumour cell and tumour microenvironment heterogeneity. Here, the authors develop a platform for the generation of patient-derived B-cell lymphoma models (termed lymphomoids) which maintain the lymphoid tissue architecture, to enable drug screening. [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]

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]

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]

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]

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]

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]

Ferocactus peninsulae (F.A.C. Weber) Britton & Rose, commonly known as the barrel cactus, is a distinctive and visually striking succulent belonging to the Cactaceae family. It is renowned for its impressive size and robust, barrel-shaped structure. F. peninsulae faces potential threats from habitat destruction and illegal collection. This study investigates the response of transverse Thin Cell Layer (tTCL) explants of F. peninsulae to various concentrations (0.5 to 2.0 mg L−1) of two cytokinins, 6-benzylaminopurine (BAP) and 6-furfuryl amino purine (Kinetin/Kin) to induce multiple shoots in vitro. The tTCL explants were inoculated on Murashige and Skoog’s medium supplemented with 1.0 mg L−1 BAP to induce fresh shoots (98.6% response). The shoots were proliferated (19.3 shoots per tTCL explant with 3.5 cm length) on a combination of BAP (0.75 mg L−1) and indole-3-acetic acid (IAA; 0.15 mg L−1). The results demonstrated the differential impact of cytokinins and auxins combinations on shoot multiplication, with BAP and IAA proving optimal for robust shoot growth while the higher concentrations led to abnormal morphogenesis and callus formation. The freshly generated shoots were rooted on 0.75 mg L−1 indole-3 butyric acid (95.8% rooting response) and hardened in the greenhouse, and the plantlets were successfully shifted to the field with a 100% survival rate. The study could help in the large-scale production of the conservation-prioritized cactus species using the tTCL culture technique.Key message: The areoles on transverse thin cell layer explants of Ferocactus peninsulae were activated using a combination of cytokinin and auxin, leading to successful shoot regeneration, rooting, and hardening of the plantlets. [ABSTRACT FROM AUTHOR]

Porcine epidemic diarrhea (PED), a contagious intestinal disease caused by the porcine epidemic diarrhea virus (PEDV), has caused significant economic losses to the global pig farming industry due to its rapid course and spread and its high mortality among piglets. In this study, we prepared rabbit polyclonal antibody and monoclonal antibody 6C12 against the PEDV nucleocapsid (N) protein using the conserved and antigenic PEDV N protein as an immunogen. A double-antibody sandwich quantitative enzyme-linked immunosorbent assay (DAS-qELISA) was established to detect PEDV using rabbit polyclonal antibodies as capture antibodies and horseradish peroxidase (HRP)-labeled 6C12 as the detection antibody. Using DAS-qELISA, recombinant PEDV N protein, and virus titer detection limits were approximately 0.05 ng/mL and 103.02 50% tissue culture infective dose per mL (TCID50/mL), respectively. There was no cross-reactivity with porcine reproductive and respiratory syndrome virus (PRRSV), porcine rotavirus (PoRV), porcine pseudorabies virus (PRV), porcine deltacoronavirus (PDCoV), or porcine circovirus (PCV). The reproducibility of DAS-qELISA was verified, and the coefficient of variation (CV) for intra- and inter-batch replicates was less than 10%, indicating good reproducibility. When testing anal swab samples from PEDV-infected piglets using DAS-qELISA, the coincidence rate was 92.55% with a kappa value of 0.85 when using reverse transcription-polymerase chain reaction (RT-PCR) and 94.29% with a kappa value of 0.88 when using PEDV antigen detection test strips, demonstrating the reliability of the method. These findings provide fundamental material support for both fundamental and practical studies on PEDV and offer a crucial diagnostic tool for clinical applications. Key points: • A new anti-PEDV N protein monoclonal antibody strain was prepared • Establishment of a more sensitive double antibody sandwich quantitative ELISA • DAS-qELISA was found to be useful for controlling the PEDV spread [ABSTRACT FROM AUTHOR]

In Brazil, around 80% of snakebites are caused by snakes of the genus Bothrops. A three-dimensional culture model was standardized and used to perform treatments with Bothrops erythromelas venom (BeV) and its antivenom (AV). The MRC-5 and L929 cell lines were cultured at increasing cell densities. Morphometric parameters were evaluated through images obtained from an inverted microscope: solidity, circularity, and Feret diameter. L929 microtissues (MT) showed better morphometric data, and thus they were used for further analysis. MT viability was assessed using the acridine orange and ethidium bromide staining method, which showed viable cells in the MT on days 5, 7, and 10 of cultivation. Histochemical and histological analyses were performed, including hematoxylin/eosin staining, which showed a good structure of the spheroids. Alcian blue staining revealed the presence of acid proteoglycans. Immunohistochemical analysis with ki-67 showed different patterns of cell proliferation. The MT were also subjected to pharmacological tests using the BeV, in the presence or absence of its AV. The results showed that the venom was not cytotoxic, but it caused morphological changes. The MT showed cell detachment, losing their structure. The antivenom was able to partially prevent the venom activities. [ABSTRACT FROM AUTHOR]

In this study, we investigated the effects of the dietary addition of lavender essential oil (LEO) on mitigating the adverse effects of high stocking densities (HSD) in the Nile tilapia, Oreochromis niloticus. We used 216 fingerlings and randomly distributed them according to a 2 × 3 factorial design, comprising two stocking densities: a low stocking density (LSD, 2.92 kg/m3) and a high stocking density (5.85 kg/m3) and three levels of LEO, 0, 0.5%, and 1%, for 70 days. We evaluated their growth, digestive enzyme activity, blood biochemical parameters, immune/antioxidant indices, and intestinal morphology. All fish were intraperitoneally inoculated with Aeromonas veronii for a bacterial challenge test at the end of the feeding trial and monitored for 14 days. After 14 days, head kidney samples were collected from all fish to assess the level of expression of immune-related and apoptosis-related genes. Additionally, tissue samples were collected from the liver, kidney, and intestine to investigate the immunohistochemistry and histopathological alterations. The results showed that the final body weight and weight gain were lower and the feed conversion ratio was higher in fish cultured under HSD compared to those cultured under LSD. The fish cultured under LSD and HSD had higher final body weight when administered dietary LEO compared to the fish that were fed the non-supplemented diet. Adding 1% LEO increased the intestinal villus height and muscular coat thickness (MT). The MT was lower in fish cultured under HSD compared to those cultured under LSD. The amylase and lipase activities were lower in the fish cultured under HSD than in those cultured under LSD. However, LEO addition increased the lipase activity in fish cultured under LSD and HSD. Fish cultured under HSD showed lower immune/antioxidant indices (lysozymes, nitric oxide, complement 3, catalase, superoxide dismutase, and glutathione-S transferase) than those cultured under LSD. However, LEO addition significantly increased the immune response and antioxidant activity in fish cultured under LSD and HSD (P < 0.01). The level of expression of the IL-1β, IL-6, IL-8, NF-kB, and TLR-4 B genes was upregulated in the LSD + LEO1, LSD + LEO0.5, and HSD + LEO1 groups. The expression of TGF-β and IL-10 was upregulated in fish that received 1% and 0.5% LEO, irrespective of whether they were cultured under LSD or HSD. The level of expression of apoptosis-related genes (caspase 3 and BAX) was upregulated in the HSD + LEO0 group and modified by the addition of LEO in their diet. The immunoexpression of NF-kB and IL-1β after bacterial challenge increased in the tissues of fish cultured under LSD and HSD after LEO was administered. These results indicated that the dietary addition of 1% LEO mitigated the harmful effects of HSD on the growth, immunity, antioxidant activity, and disease resistance of the Nile tilapia. [ABSTRACT FROM AUTHOR]

Key message: The Agrobacterium-mediated transformation system of three elite indica rice varieties was explored and two varieties yielded positive transgenic plants. Fourteen elite indica rice varieties with different genotypes were selected for callus induction, and varieties NY49, KW and CL38 were chosen for the establishment of the Agrobacterium transformation system because of their excellent tissue culture characteristics. The callus induction frequency was the highest when the concentration of 2,4-Diphenoxyacetic acid (2,4-D) was respectively at 2.5 mg/L, 3.0 mg/L and 2.8 mg/L for NY49, KW and CL38 after optimizing the tissue culture conditions. Moreover, medium types of CIM1, CIM8 and CIM7 were most suitable for the varieties NY49, KW and CL38, respectively, with high callus induction frequency. The regeneration frequency of CL38 and NY49 was as high as 73.33% and 50.00%, respectively, in MS medium containing 2 mg/L 6-benzyladenine (BAP), 2 mg/L kinetin (KT) and 0.5 mg/L naphthylacetic acid (NAA). Finally, transgenic seedlings of CL38, NY49 and NIP were obtained with infection of Agrobacterium EHA105 carrying the pYLCRISPR/Cas9PUB-H vector. The successful editing frequency detecting by sequencing in CL38 and NIP were 62.50% and 77.78%, respectively. The establishment of the transformation system provides a useful tool for gene function study and gene editing breeding in indica rice cultivars. [ABSTRACT FROM AUTHOR]

The study investigates the impact of polyethylene glycol (PEG), sodium nitroprusside (SNP), and different carbon sources on the in vitro callus induction and regeneration of drought-tolerant and drought-sensitive rice (Oryza sativa L.) genotypes. PEG, a known osmotic agent, mimics drought conditions, thus providing insights into the plant's response mechanisms under stress. SNP, a nitric oxide donor, is evaluated for its role in modulating stress responses and promoting growth under adverse conditions. Various carbon sources are evaluated to determine their efficacy in supporting callus induction and subsequent regeneration. Results indicate differential responses between drought-tolerant and drought-sensitive rice genotypes, highlighting the potential for optimizing tissue culture protocols to enhance the resilience and regenerative capacity of rice under drought stress. While moderate PEG (5%) stimulated callus induction and regeneration, higher concentrations resulted in adverse effects, including reduced callus mass and regeneration potential. Morphological changes were also observed in response to PEG concentrations, highlighting genotype-specific responses. Meanwhile, combined effect of carbon sources such as maltose and sorbitol along with nitrogen sources "SNP" enhanced callus induction (> 90%) and regeneration (> 90%) under 20% PEG stress in both drought-tolerant and drought-sensitive rice. [ABSTRACT FROM AUTHOR]

Purpose: Infectious spondylitis is caused by hematogenous seeding or adjacent soft tissue infection. No study has provided evidence that incubating biopsy specimens in blood culture bottles could enhance detection rates, nor has any study compared this method with conventional culture techniques. We aimed to assess the diagnostic yield of open microsurgical biopsies for infectious spondylitis and the efficacy of various culture media in the presence and absence of pre-biopsy antibiotic therapy. Methods: This retrospective study, which was conducted at a university-affiliated teaching hospital in Korea, enrolled 165 adult patients with suspected infectious spondylitis between February 2014 and September 2020. The diagnostic yield of open biopsy was compared among three culture media, namely, blood culture bottles, swab culture using transport media, and tissue culture using plain tubes, while considering preoperative antibiotic exposure. Results: Causative bacteria were identified in 84.2% of all cases. Blood culture bottles had the highest positivity rate (83.5%), followed by swab cultures (64.4%) and tissue cultures (44.9%). The differences in positivity rates were significant (P < 0.001). Preoperative antibiotic therapy reduced detection rates across all media, particularly in tissue cultures. Conclusions: We established the high diagnostic yield of open microsurgical biopsy using blood culture bottles, suggesting that pre-biopsy antibiotic therapy significantly affects bacterial detection, thereby underscoring the importance of culture medium selection in the diagnosis of infectious spondylitis. [ABSTRACT FROM AUTHOR]

Handroanthus chrysanthus Jacq. S. O. Grose and Tabebuia rosea (Bertol.) Bertero ex A.DC are two forest species that grow in the coastal region of Ecuador and are threatened with extinction. A protocol for the mass multiplication of these species was developed using in vitro culture techniques. The cultures were initiated from seeds, and the effect of two culture media: Woody Plant Medium (WPM) and Murashige-Skoog (MS), two concentrations of NaClO (0.5% and 1%), and two disinfection times (3 and 5 min) was studied. During multiplication, the effect of three concentrations of two cytokinins: 6-benzylaminopurine (6-BAP) 4.4, 5.5, or 6.6 μM; kinetin 4.6, 5.75, or 6.9 μM) on the number of shoots, their length, and diameter was analyzed. This phase of the experiment was carried out in two successive multiplications. For rooting, two concentrations of indole-3-butyric acid (IBA) (2.45 and 4.9 μM) were tested, and the number of roots formed and their length were determined. It was demonstrated that the WPM medium is the most suitable for the in vitro culture of both species and that disinfection time and NaClO concentration affect each species differently. For the multiplication of H. chrysanthus, the most suitable cytokinin was 6-BAP 6.6 μM; T. rosea performed better in the absence of cytokinins. IBA 2.45 μM produced the best results for the rooting of H. chrysanthus, while for T. rosea, IBA 4.9 μM was the most suitable. The acclimatized plants showed a high survival rate, demonstrating the feasibility of using this methodology for the accelerated propagation of these endangered species. Key message: In this research, the culture medium requirements and conditions for the micropropagation of H. chrysanthus and T. rosea were fine-tuned. This technique can be implemented to obtain plants for use in reforestation. [ABSTRACT FROM AUTHOR]

The present study aimed to enhance salicin and direct regeneration in willow (Salix safsaf Forssk) using the sodium nitroprusside (SNP) regulation of nitric oxide (NO) and vinasse for its nutrition effect in culture medium. Internodes of Salix safsaf were cultured on Murashige and Skoog (MS) medium supplemented with benzyl adenine (BA) (0.25 mg L−1) and different concentrations of SNP (0, 5, 10, 15, and 20 mg L−1) or vinasse (0, 5, 10, and 20%) to examine shoot regeneration, antioxidant defense enzyme activity, total phenolic compounds, flavonoids, and salicine contents. The reported data revealed that application of SNP at 15 mg L−1 and vinasse at 10% induced a significant effect in vitro Salix safsaf shoot regeneration. To confirm that, nitric oxide is required for auxin-mediated activation of cell division in a dose-dependent manner. A concentration of 15 mg L−1 SNP promotes regeneration and salicin accumulation (3162.16 mg/100 g) during signaling action. On the other hand, the cross talk effect of nitric oxide and vinasse combination in Salix safsaf significantly induced a synergistic effect on direct propagation more than vinasse alone. SNP significantly stimulates salicylate accumulation in a dose-dependent manner, but the data on the association of vinasse and SNP on salicylate up-regulation showed a significant reduction in salicin accumulation when SNP was combined with 10% vinasse, which directly affected the signaling action of SNP as secondary product stimulators. Vinasse's phenolic compounds affect directly on the reduction activity of SNP to suppress its signaling action, or indirectly by inhibiting the sequence cascade of the SNP signaling transduction process to decrease the accumulation of salicin contents. Data confirmed that vinasse and SNP stimulated the antioxidant enzymes activity throw quenching the stimulated reactive oxygen species that produced via SNP. Results show that modified media with SNP administration at 15 mg L−1 and the combination of vinasse at 10% and SNP at 15 mg L−1 are recommended for modifying tissue culture media for induced direct regeneration and salicin accumulation in tissue culture applications, which will be very useful for commercial salicin overproduction as a biological active ingredient in willows. [ABSTRACT FROM AUTHOR]

Recently, it was reported that a testicular organ culture system (TOCS) using polydimethylsiloxane (PDMS) chips with excellent oxygen permeability and biocompatibility, called the PDMS-chip ceiling (PC) method, enables improved spermatogenesis efficiency. We investigated whether this PC method is useful for detecting impaired spermatogenesis caused by busulfan (Bu), a typical testicular toxicant. In this study, testicular tissue fragments from Acro3-EGFP mice, which express the green fluorescent protein (GFP) and reflect the progression of spermatogenesis, were subjected to the PC method. When treated with Bu, cultured tissues shrank in volume, and their GFP-expressing area decreased or disappeared. Histological examination confirmed the regression of spermatogenesis. In addition, immunohistochemical examination revealed that spermatogonia, including spermatogonial stem cells (SSCs), were the primary targets of Bu toxicity. Time-course analysis demonstrated that the recovery of spermatogenesis, dependent on Bu concentration, correlated closely with the severity of damage to these target cells. These results suggest that the PC method is a useful approach for detecting spermatogenesis impairment accurately through faithful recapitulation of spermatogenesis in vivo. [ABSTRACT FROM AUTHOR]

The emerging field of biorefinery presents an opportunity to extract valuable resources from plant biomass like hemp, while tissue culture techniques, such as micropropagation, can produce uniform and high-quality plant material for biorefinery purposes. This study evaluated the use of hormonal and organic diets in creating callus from leaf fragments in three medical hemp cultivars over 8 weeks for biorefinery applications. The most effective treatment was the modified MS diet supplemented with 0.5 mg/L 2,4-D plus 0.5 mg/L KN, which induced callus formation in all three cultivars with an average size of 2.1 cm. Adding 150 mg/L of coconut water to the MS formula also showed significant results, inducing up to 50% callus formation in all three cultivars with an average size of 1.3 cm. Three different photoperiods were tested, with the 16:8 photoperiod being the most effective. White light was the most effective color, followed by blue light. The highest yield of extract was obtained from the callus section of Carmagnola hemp in the MS formula without growth regulators, with a yield of 2.75%. Consequently, this study provides valuable insights into the use of hormonal and organic diets, photoperiod, and light color for callus formation from leaf fragments in medical hemp, which could have important implications for producing high-value compounds via biorefinery processes. [ABSTRACT FROM AUTHOR]

Positive allosteric modulators (PAMs) are of great interest to compensate for the lack of NMDA receptor function in the CNS in dementia. Known PAMs increase the amplitude of whole-cell ionic currents carried by NMDA receptors, but do not affect their calcium-calmodulin dependent desensitization. We studied the possibility of modulating NMDA receptor desensitization by the recently synthesized PAM GNE-9278, which has a unique binding site on the transmembrane domain. Experiments were performed on native NMDA receptors expressed in rat cortical neurons in primary tissue culture. Using the method of "patch-clamp" recording of transmembrane currents, a comparative study of the effect on NMDA receptor desensitization of three substances potentiating NMDA receptor currents was carried out: GNE-9278 (10 µM), dithiothreitol (1 mM) and copper ions (5 µM). These substances increased the amplitude of currents evoked by 100 µM NMDA, but only GNE-9278 reduced the difference between equilibrium and peak current amplitudes by 15%. In addition, GNE-9278 doubled the time constant of the decline from peak to equilibrium state thus attenuating NMDA receptor desensitization. Because GNE-9278 did not alter the effective concentration of extracellular calcium to induce desensitization, its effect probably does not interfere with receptor-calmodulin interaction. Analysis of the shape of the currents within the kinetic model showed that GNE-9278 reduced two kinetic parameters: the channel closure rate, which determines the open state time, and the rates of receptor entry into and exit from the desensitized state, which determines the probability of the open state of the channel. Modulation of calcium-dependent NMDA receptor desensitization places GNE-9278 apart from other known PAMs, which is likely determined by the GNE-9278 binding site in the pre-M1 segment of the GluN1 subunit. [ABSTRACT FROM AUTHOR]

This review explores recent advances in the biotechnology of Lamprocapnos spectabilis (L.) Fukuhara (commonly known as bleeding heart), a valuable ornamental-medicinal perennial. The article covers in vitro morphogenesis, cryopreservation techniques, and methods for inducing variability. The establishment of in vitro cultures utilized Murashige and Skoog medium enriched with various auxins, cytokinins, gold nanoparticles, and plant extracts, under both fluorescent and wide-spectrum LED lighting. Axillary bud activation and indirect somatic embryogenesis were more efficient, particularly in the presence of kinetin and picloram, respectively, compared to adventitious shoot regeneration. Significant cultivar differences were observed, with ‘Valentine’ being the easiest and ‘White Gold’ the most challenging to culture in vitro. To mitigate stress caused by classical growth regulators, alternative substances such as nanoparticles and natural extracts were used. Gold nanoparticles enhanced shoot proliferation and plantlet quality, while coconut and rice extracts improved survival rates during acclimatization. Enhanced metabolite production was achieved using exogenous auxins and gold nanoparticles. Guaiacol peroxidase was identified as a sensitive oxidative stress marker, with glutathione reductase being the most stable under stress. Cryogenic techniques incorporating explant encapsulation, i.e. encapsulation-vitrification, showed high effectiveness and genetic stability of plants, with nanomaterials boosting effectiveness. Coconut extract also enhanced post-thaw shoot proliferation, while sesame extract served as a natural retardant for slow-growth cultures. Mutagenic effectiveness ranked as microwaves < nanoparticles < X-rays. Comprehensive genetic variability insights were provided by integrating multiple SPAR marker systems. This review underscores the promising biotechnological advancements for L. spectabilis, emphasizing the potential of in vitro techniques, innovative cryopreservation methods, and the application of nanoparticles and plant extracts to enhance micropropagation, genetic variability, and metabolite production, thereby contributing to the conservation and commercial sustainability of this valuable ornamental-medicinal perennial. [ABSTRACT FROM AUTHOR]

Prenatal exposure to toxins can adversely affect long-term health outcomes of the offspring. Though chemotherapeutics are now standard of care for treating cancer patients during pregnancy, certain compounds are known to cross the placenta and harm placental tissue. The consequences for the fetus are largely unexplored. Here we examined the responses of newborn cord blood mononuclear cells in tissue culture to two chemotherapeutic drugs, cyclophosphamide and epirubicin, when either directly exposed to these drugs, or indirectly after crossing a placenta trophoblast bilayer barrier. Cord blood mononuclear cells exposed to the conditioned media obtained from cyclophosphamide-exposed trophoblast barriers showed a significant 2.4-fold increase of nuclear ROS levels compared to direct exposure to cyclophosphamide. Indirect exposure to epirubicine-exposed trophoblast barriers not only enhanced nuclear ROS levels but also significantly increased the fraction of cord blood cells with double strand breaks, relative to directly exposed cells. Neither apoptosis nor proliferation markers were affected in cord mononuclear blood cells upon direct or indirect exposure to cyclophosphamide or epirubicin. Our data suggests that trophoblast cells exposed to cyclophosphamide or epirubicine may induce an indirect 'bystander' effect and can aggravate genotoxicity in the fetal compartment. [ABSTRACT FROM AUTHOR]

Introduction: Although antifungal supplementation reduces the fungal load in the corneal storage medium, consensus is lacking on the influence of dosing and temperature. The study aims to evaluate the impact of eye bank warming protocol and timing of antifungal supplements on efficacy in Optisol-GS and tissue. Methods: Corneoscleral rims contaminated with Candida albicans (C. albicans) were incubated in Optisol-GS, either without antifungal agents or with the addition of amphotericin B or voriconazole at various concentrations (2 ×, 5 ×, 10 ×, and 20 × MIC), at different time points, and under various preservation temperatures (2–8 °C versus 2 h-room temperature exposure). Antifungal efficacy was evaluated by counting viable yeast colonies cultured from Optisol-GS samples. Tissue sterility was determined through direct tissue culture and histological examination of the contaminated rims after a 14-day incubation period. Results: Room temperature exposure did not increase colony growth at the same multiple MIC of antifungal agents. Although antifungal addition reduced C. albicans growth in a concentration-dependent manner, yeast growth was still observed in all Optisol-GS samples with a single supplementation after a 14-day incubation. Only groups with additional antifungal supplementation on either day 2 or day 6 showed a 99% or greater reduction of C. albicans growth in Optisol-GS samples and yielded negative results in direct tissue culture. Conclusions: The eye bank warming protocol did not compromise antifungal efficacy. To sustain the required concentration and effectively reduce C. albicans growth in Optisol-GS and contaminated tissue, additional antifungal supplementation on either day 2 or day 6 was necessary during a 2-week preservation period. [ABSTRACT FROM AUTHOR]

Juglans mandshurica is an important component of Korean pine mixed and secondary forests in Northeast China and is also a precious timber and economic species. In this study, plant regeneration of J. mandshurica has been established using stems with bud and mature zygotic embryos as explants. The results showed that placing the stems with buds in low-temperature conditions (0 to 4°C) and in darkness for 5 d inhibited the browning of the explants and promoted bud sprouting; Na2S2O3 also inhibited the browning of mature zygotic embryos significantly. The suitable basal media and hormones for bud sprouting and zygotic embryo germination were (1) WPM medium (woody plant medium) containing 2.0 mg·L−1 6-benzylaminopurine (6-BA) and 0.01 mg·L−1 3-indolebutyric acid (IBA) and (2) DKW medium (Driver and Kuniyuki medium) containing 2.5 mg·L−1 6-BA and 0.1 mg·L−1 IBA. The suitable basal medium and hormone for rooting of shoots from buds was 1/2 Murashige and Skoog (MS) medium (the macro-elements were half-strength of MS medium) supplemented with 1.0 mg·L−1 IBA, and the suitable basal medium and hormone for rooting of seedlings derived from zygotic embryo was 1/2 MS medium containing 1.0 mg·L−1 IBA, and DKW medium with 2.5 mg·L−1 6-BA, 0.1 mg·L−1 IBA, 0.2 mg·L−1 gibberellin A3 (GA3), and 0.2 g·L−1 activated carbon (AC). About 95% of the rooted plantlets survived after acclimation and transplantation. These results provided a foundation for the scale propagation to meet the demands of forestry production of the species. [ABSTRACT FROM AUTHOR]

Axonopus compressus (Swartz) Beauv., a perennial herb in the Poaceae that has been introduced to a number of tropical and subtropical countries and regions, can serve as a lawn ground cover while its leaves can be developed as a bioenergy resource. It also displays some resistance to heavy metals, allowing it to be planted in areas and urban green spaces polluted by heavy metals. A. compressus is also used as a traditional Chinese medicine. This study is the first report on tissue culture of A. compressus. Stem explants induced shoot clusters on 6-benzyladenine (BA)-containing Murashige and Skoog (MS) medium, with 2.0 mg L–1 BA inducing a shoot proliferation coefficient (SPC) of 10.89 within 30 d, while MS medium containing 2.0 mg L–1 BA and 0.1 mg L–1 α-naphthaleneacetic acid (NAA) amplified SPC to 12.88 within 30 d. SPC on MS medium supplemented with kinetin (KIN) or thidiazuron (TDZ) was not as high as on medium with BA and never exceeded 3.57 within 30 d. A. compressus formed adventitious roots easily, within 15 d, most efficiently on ½MS medium supplemented with 0.1 to 0.5 mg L–1 IAA, NAA, or IBA, or even on auxin-free medium. Resulting plantlets displayed a high survival rate (> 98%) when transplanted to a substrate containing humus, or humus mixed with vermiculite or sand. Callus was also successfully induced from leaf sheaths on MS medium containing 1.0 mg L–1 2,4-dichlorophenoxyacetic acid, even more so when 1.15 g L–1l-proline was added. Induced callus was able to differentiate into adventitious shoots on MS medium with 0.1 to 0.5 mg L–1 KIN or BA. An efficient system of shoot proliferation, callus induction, and regeneration in A. compressus has been established. [ABSTRACT FROM AUTHOR]

Dendrobium species, esteemed as a group of orchid plants with important ornamental and medicinal value, are currently experiencing significant developmental advancements. This research encompasses a comprehensive review of Dendrobium tissue culture literature spanning 2008 to 2023, sourced from the Web of Science Core Collection database. Employing the bibliometric analysis tool CiteSpace, we conducted a visualization and presentation of data encompassing research institutions, authorship patterns, key terminology, and emergent themes, elucidating the current status, hotspots, and frontiers of Dendrobium tissue culture research. After thorough screening, a total of 261 papers were included in this study. The publication count exhibited a consistent annual increase. Chinese Acad Sci, Univ Sains Malaysia, North Eastern Hill Univ, Univ Debrecen, and Univ Putra Malaysia emerged as primary research institutions, while a core team led by J.A. Teixeira da Silva, S. Sreeramanan, S. Kumaria, and N. Chen played a dominant role. Central research themes revolve around optimizing the tissue culture system of medicinal Dendrobium, and exploring the genetic stability and gene function. Furthermore, the field is witnessing a trajectory towards future multidisciplinary cross-collaboration. [ABSTRACT FROM AUTHOR]

Cibotium barometz (L.) J. Sm., a fern, holds significant value in traditional Chinese medicine. This research aimed to establish a rapid tissue breeding technology for Cibotium barometz (L.) and provide a theoretical basis for its artificial breeding and industrial planting. The optimum conditions for spores germination were selected, including both the best sterilization time and the ideal spore germination medium. Subsequently, the optimal medium for proliferation and differentiation were chosen, and the optimal concentration of plant hormones was screened to determine the optimal medium. The rooting experiment was performed in two single-factor experiments by using differentiated sporophytes as materials. These experiments showed that the optimal sterilization time was 11 min and the best medium for germination was 1/8MS + 0.02 mg/L IAA. MS medium was suitable for proliferation, while WPM medium was suitable for sporophyte differentiation. The best medium for growth was MS + 1.0 mg/L IAA + 0.5 mg/L 6-BA. In the absence of sucrose, the seedlings barely grew and failed to differentiate into sporophytes. They showed optimal growth and differentiation at a sucrose concentration of 20 g/L. The plant growth regulators plant growth regulators used, including IAA, NAA, 6-BA, and GA3, showed more effective differentiation, wherease KT had a slightly weaker effect. The rooting rate reached the maximum when the concentrations of ABT and IBA were at 1.5 mg•L− 1 and IAA was at 1.0 mg•L− 1. The IBA had a significant effect on the average and the longest root length, while the three PGRs had no obvious effect on the average or maximum diameter. Plant tissue culture seedlings still have good genetic stability after subsequent generation. In this study, the optimal cultivation conditions for rapid breeding technology in tissue culture, covering stages from germination and growth to differentiation and rooting, were obtained. Moreover, sporophytes were successfully transplanted to the external environment. This study provided a theoretical reference for the industrial planting of C. barometz.Key message: In this study, a more systematic, complete and efficient tissue culture rapid propagation system was formed from spore germination, proliferation, differentiation, and took roots, along with the sporophytes for transplanting and culture. [ABSTRACT FROM AUTHOR]

Neolamarckia cadamba (N. cadamba) is an evergreen tree species known for its rapid growth, remarkable wood properties, and significant value in medicine, feeding, and landscape. In order to clone a N. cadamba individual with excellent genotype, a plant regeneration protocol was successfully established with leaves wrapped by bud scales as explants. The optimal sterilization method for the leaves was 0.1% Mercury Chloride (HgCl2) treatment for 1 min before culturing on Murashige and Skoog’s medium (MS) supplemented with 3.0 mg/L Thidiazuron (TDZ), 0.1 mg/L 2–4 Dichlorophenoxyacetic acid (2-4D), 0.05 mg/L α-Naphthaleneacetic acid (NAA) and 1 mL/L Plant Preservative Mixture (PPM) to induce calluses. The medium containing 1 mL/L PPM could effectively inhibit explant contamination without an unfavorable impact on the final induction rate of callus from the leaves. Three types of calluses were induced from the leaves cultured on the above medium. Among them, only the Type II callus, which was green and nodular, had few particle masses, could differentiate into adventitious shoots on the MS medium supplemented with 1.5 mg/L 6–Benzylaminopurine (6-BA) and 0.05 mg/L NAA, with the induction rate of 78.89% and adventitious shoot number per callus of 11.67. The adventitious shoots were proliferated on the MS medium supplemented with 1.0 mg/L 6-BA and 0.05 mg/L Indole-3- butyric acid (IBA) with the proliferation coefficient of 3.37. And the micro-shoots developed roots in the MS medium supplemented with 0.05 mg/L NAA and 0.05 mg/L IBA. The regeneration protocol can be used in the propagation and large scale production of seedlings with the same genotype as an excellent individual of N. cadamba in the field.Key message: A plant regeneration protocol was successfully established with leaves wrapped by bud scales in the field, furthermore it can be used in the propagation and large scale production of seedlings with the same genotype as an excellent individual of N. cadamba. [ABSTRACT FROM AUTHOR]

The emerging field of cellular agriculture has accelerated the development of cell-cultivated adipose tissue as an additive to enhance the flavor of alternative meat products. However, there has been limited research to evaluate the sensory profile of in vitro-grown tissues compared to conventionally obtained animal fat. This study aimed to investigate the aromatic characteristics of cell-cultivated fat tissue as a flavor enhancer for meat alternatives. Porcine dedifferentiated fat (PDFAT) cells were clonally isolated and differentiated into adipocytes. This cultured adipose tissue was then analyzed alongside native porcine fat using gas chromatography-mass spectrometry (GC/MS) coupled with descriptive sensory analysis by human consumers. This evaluation enabled quantitative and qualitative assessments of volatile compounds released during cooking for both in vitro and in vivo porcine fats. The volatile profiles generated during the cooking process and fatty aroma characteristics reported by sensory consumers were largely similar between the two fat sources, with some differences in select compounds and aroma attributes. Ultimately, the consumers found comparable overall liking scores reported between the conventional and cultured porcine fats. These findings provide valuable sensory evidence supporting the viability of cell-cultivated adipose tissue as a flavor component of meat alternatives, substituting for conventional animal fat. [ABSTRACT FROM AUTHOR]

Previously, we reported successful cellular expansion of a murine colorectal carcinoma cell line (CT-26) using a three-dimensional (3D) engineered extracellular matrix (EECM) fibrillar scaffold structure. CCL-247 were grown over a limited time period of 8 days on 3D EECM or tissue culture polystyrene (TCPS). Cells were then assayed for growth, electroporation efficiency and Vigil manufacturing release criteria. Using EECM scaffolds, we report an expansion of CCL-247 (HCT116), a colorectal carcinoma cell line, from a starting concentration of 2.45 × 105 cells to 1.9 × 106 cells per scaffold. Following expansion, 3D EECM-derived cells were assessed based on clinical release criteria of the Vigil manufacturing process utilized for Phase IIb trial operation with the FDA. 3D EECM-derived cells passed all Vigil manufacturing release criteria including cytokine expression. Here, we demonstrate successful Vigil product manufacture achieving the specifications necessary for the clinical trial product release of Vigil treatment. Our results confirm that 3D EECM can be utilized for the expansion of human cancer cell CCL-247, justifying further clinical development involving human tissue sample manufacturing including core needle biopsy and minimal ascites samples. [ABSTRACT FROM AUTHOR]

Multiplexed in situ RNA imaging offers new opportunities for gene expression profiling by providing high-throughput spatial information. In this work, we present a cyclic combinatorial fluorescent in situ hybridization (combFISH) assay to achieve multiplexed detection of RNA in cell cultures and tissues. Specifically, multiplexing is achieved through cyclic interrogation of barcode sequences on the rolling circle amplicons generated from the padlock probe assay by using sets of combinatorial detection probes. Theoretically, combFISH can detect 64 genes in three hybridization cycles by combinatorial barcoding using 12 fluorescently labeled detection probes. Our method eliminates sequencing-by-ligation (SBL) chemistry in the in situ sequencing protocol and directly uses RNA as targets for ligation, making it more straightforward. We showed that our method works in fresh-frozen and formalin-fixed paraffin-embedded tissue sections. With its straightforward protocols, we expect our method to be adopted by the scientific community and extended to clinical settings. Multiplexed RNA in situ imaging by combinatorial FISH decoding [ABSTRACT FROM AUTHOR]

Citrus is one of the most important global fruit crops, with oranges accounting for more than half the total production. Citrus can be genetically engineered for improved traits, but the process is severely limited by shoot regeneration rates, especially in commercial varieties. Although standardizing a transformation protocol is difficult due to genotype and season dependencies, use of an optimal mineral nutrient basal culture medium increases the success of recovering transgenic Citrus cells. The purpose of this study is to improve shoot regeneration from juvenile tissue in multiple Citrus genotypes. Explants of 11 Citrus cultivars were regenerated in 21 published media to determine the optimal media for each genotype. The number of shoots from juvenile tissue were counted 90 days after the explants were first cultured. The type of basal medium strongly affected the rates of shoot regeneration from Citrus juvenile tissue, and the effectiveness was negatively correlated with the level of ammonium. For each Citrus grouping, optimized media formulations were determined, and potential improvements were predicted. In general, medium R7100 appeared to be the most effective for regeneration of the Citrus genotypes tested.Key Message: Results from this study indicate that basal medium strongly affected the rates of shoot regeneration from citrus juvenile tissue, and the effectiveness was negatively correlated with the level of ammonium present. [ABSTRACT FROM AUTHOR]

Crocus scepusiensis (Rehm. & Woł.) Borbás ex Kulcz., a critically endangered herbaceous plant which serves as a valuable source of bioactive compounds found across Europe and Asia. The aim of this study was to produce a calli from two different plant parts (leaf and shoot tip) for the critically endangered C. scepusiensis through tissue culture techniques, characterize the resulting calli through chemical profiling, with a focus on identifying key phytoconstituents, and lay the groundwork for future research on the biological activities of these calli extracts. Leaf disc and micro shoot tip explants were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of 6-benzylaminopurine (BA) and α-naphthaleneacetic acid (NAA) to induce organogenic calli. The resulting calli exhibited distinct biochemical profiles. Moreover, a phytochemical analysis was conducted to compare the metabolite composition of callus 1 (derived from leaf discs) and callus 2 (derived from shoot tips). Callus 1 displayed a higher total phenolic content (30.3558 ± 1.3564 mg (GAE)/g) compared to callus 2 (29.1543 ± 0.9754 mg (GAE)/g). Similarly, callus 1 exhibited a greater total flavonoid content (26.0089 ± 1.8029 mg (RE)/g) than callus 2 (18.4464 ± 1.4797 mg (RE)/g). Liquid chromatography-photodiode array-electrospray ionization-tandem mass spectrometry (LC-PDA-ESI-MS/MS) analysis revealed the presence of 26 and 25 constituents in callus 1 and 2, respectively. Fourteen and thirteen of these identified compounds have been previously reported in other Crocus species, with 22 constituents common to both calli. Twelve constituents were reported here in Crocus for the first time as far as we know.Key message: Tissue culture successfully produced Crocus scepusiensis calli with distinct phytoconstituents profiles, potentially offering novel sources of bioactive compounds. [ABSTRACT FROM AUTHOR]

Tissue culture is preferred for solving the shortcomings of low efficiency of conventional propagation in tree peony (Paeonia sect. Moutan), an economically important woody plant in China with various uses. Paeonia ostii ‘Feng Dan’ is one of the varieties with strong adaptability and high oil productivity. However, in vitro regeneration via organogenesis is hard to obtain. For instance, the indirect organogenesis from callus is a lengthy process, and the direct organogenesis is lacked. This study presents a protocol of direct organogenesis using cotyledon explant for in vitro regeneration of P. ostii ‘Feng Dan’, and includes two pathways. The explants need pretreated in the optimal dedifferentiation induction medium (DIM) [Murashige and Skoog (MS) medium with 2.27 µM thidiazuron (TDZ) + 5.37 µM α-naphthylacetic acid (NAA)] for 10 days, and then the cotyledons without callus induced were transferred to differentiation medium (DM) with 6 subcultures, 90 days in total. The best DM for the frequency of leaf clusters induction (66.67%) was Woody plant medium (WPM) containing 4.04 µM N-(2-chloro-4-pyridyl)-N-phenylurea (CPPU) + 4.54 µM TDZ, and for the frequency of MNs induction (41.67%) was WPM containing 2.02 µM CPPU + 2.27 µM TDZ. For two pathways, mean number of regenerated shoots (≈ 4 shoots per explant), the frequency of root induction (≈ 50%) and the frequency of survival (≈ 40%) were similar. Histological studies confirmed the process of direct organogenesis, and revealed that shoots and MNs were originated from the increased division of meristematic cells under cortical tissue, as well as from actively dividing meristematic cells around the vascular center. Moreover, shoots regenerated through MNs differentiation were originated from the epidermal and subepidermal cells. This study is supplements in the field of in vitro regeneration in tree peony, and is an improvement to current protocols of clonal micropropagation, at the same time facilitating fundamental studies of developmental biology and genetic transformation.Key message: Plant regeneration through two pathways of direct organogenesis using cotyledon explant was first presented in Paeonia ostii ‘Feng Dan’. [ABSTRACT FROM AUTHOR]

Main conclusion: Wheat lines harboring wild-relative chromosomes can be karyotypically unstable during long-term maintenance. Tissue culture exacerbates chromosomal instability but appears inefficient to induce somatic homoeologous exchange between alien and wheat chromosomes. We assessed if long-term refrigerator storage with regular renewal via self-fertilization, a widely used practice for crop germplasm maintenance, would ensure genetic fidelity of alien addition lines, and explored the possibility of inducing somatic homoeologues exchange by tissue culture. We cytogenetically characterized sampled stock seeds of originally confirmed 12 distinct wheat–Thinopyrum intermedium alien addition lines (dubbed TAI lines), and subjected immature embryos of the TAI lines to tissue culture. We find eight of the 12 TAI lines were karyotypically departed from their original identity as bona fide disomic alien addition lines due to extensive loss of whole-chromosomes of both Th. intermedium and wheat origins during the ca. 3-decade storage. Rampant numerical chromosome variations (NCVs) involving both alien and wheat chromosomes were detected in regenerated plants of all 12 studied TAI lines, but at variable rates among the wheat sub-genomes and chromosomes. Compared with NCVs, structural chromosome variations (SCVs) occurred at substantially lower rates, and no SCV involving the added alien chromosomes was observed. The NCVs manifested only moderate effects on phenotypes of the regenerated plants under field conditions. [ABSTRACT FROM AUTHOR]

The high cost of establishing a cell and tissue culture facility has denied students an opportunity to practice proper aseptic techniques during their studies. An in-house application named AsepticTech Virtual Reality (VR) was developed to simulate a cell and tissue culture facility for students to train their aseptic techniques virtually. However, the performance gain associated with the application will be limited by the student's willingness to use the application. Therefore, this study aims to investigate the driving factors behind students' intention to use AsepticTech VR. A total of 55 biomedical undergraduates were granted access to AsepticTech VR. After using the application, the students completed a Technology Acceptance Model (TAM) questionnaire. It measures seven latent constructs that are believed to influence the students' behavioral intention to use the application, namely Perceived Usefulness (PU), Perceived Ease of Use (PEU), Attitude Towards Use (ATU), Behavioral Intention (BI), Perceived Enjoyment (PENJ), Perceived Health Risk (PHR) and Self-Efficacy (SE). The questionnaire was preliminarily evaluated on its validity and reliability using Confirmatory Factor Analysis (CFA), Average Variance Extracted (AVE), Composite Reliability (CR), and Cronbach's alpha. Subsequently, the interactions between the seven latent variables were analyzed via path analysis. Findings of the path analysis suggested that ATU is the most influential factor on BI, followed by PENJ and SE. In turn, ATU was positively and significantly influenced by PENJ, whereas PHR exerted a significant negative influence on ATU. This study revealed that enjoyment and comfort are the principal factors influencing students' acceptance of AsepticTech VR. This study also identifies other determinants that influence users' acceptance of AsepticTech VR and paves guidance for the future development of the application. [ABSTRACT FROM AUTHOR]

Growth performance (specific growth rate [SGR] and biomass yield), carrageenan properties (yield, viscosity, and gel strength), and disease and pest incidence were compared between Kappaphycus alvarezii var. tambalang (brown) tissue-cultured (TC) plantlets and farm-sourced (FS) seedlings. Seedlings used in the trials were propagules (4‒19 g pieces) and these were deployed on lines in sea cages over three separate runs of 60 days. The highest growth rate was recorded in TC plantlets and the SGRs were significantly higher (1.5‒6.5x) in TC than FS after Day of Culture (DOC) 15 (i.e., measurements taken every 15 days in each cycle). The carrageenan properties did not differ between treatments, with the exception of viscosity at DOC 45, where TC was higher than FS. "Ice-ice" disease (IID) was observed in both FS and TC in May‒June and August‒October at DOC 45 to 60, coinciding with changes in environmental conditions (i.e., dry‒wet and wet seasons). IID in FS had a moderate to very high incidence (31.2‒86.2%), while TC IID had a lower range of incidence (12.8‒60.6%). On the other hand, epiphytic filamentous algae (EFA) were observed only in August‒October with a low incidence (≤25%). However, EFA appeared earlier in FS than in TC. Taken together, these results highlight that tissue-cultured seaweeds had higher growth performance without compromising carrageenan quality and may be more resistant to health problems than farm-sourced seedlings. Studies should now assess the long-term viability of using tissue-cultured seaweeds over farm-sourced seedlings scaling to allow testing in standard farm settings, including the size of the initial seed. [ABSTRACT FROM AUTHOR]

Multidrug resistant strains and fungi add to treatment conundrums in skull base osteomyelitis (SBO). Deep tissue culture in these patients is challenging due to their advanced age and co-morbidities. Besides, fungal culture positivity is seen only in 60% of invasive aspergillosis. To determine the efficacy of a minimally invasive test—Serum Galactomannan (sGM)–for diagnosing fungal SBO. Prospective observational study. Thirty- three patients, clinically diagnosed with SBO were included in this study. Baseline ESR (Erythrocyte Sedimentation Rate), CRP (C- Reactive Protein), pain score, and sGM were noted for all patients. Antifungal Voriconazole was initiated on patients if the sGM values were more than 0.8. At the 12th week of treatment, all parameters were repeated and compared with the baseline values. A significant reduction was noted in ESR, CRP, and pain scores at the 12th week of treatment compared to the baseline values in patients with raised sGM values who were started on Voriconazole. For a culture-proven fungal skull base osteomyelitis with a cut-off value of sGM > / = 0.8, the obtained sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy were 81.82, 36.36, 39.13, 80 and 51.52% respectively. At a cut-off value of sGM > / = 01.6, the values of sensitivity/specificity, PPV, NPV and accuracy were 81.82/72.73%, 60%, 88.89% and 75.76%. Culture-negative patients in SBO with sGM value > 0.8 were more likely to be fungal SBO. An sGM cut-off of 1.6 was observed to give maximum accuracy for diagnosing fungal SBO. [ABSTRACT FROM AUTHOR]

Waterhemp (Amaranthus tuberculatus (Moq.) Sauer) is one of the most problematic weeds in the Midwestern agricultural systems in the USA. As a consequence of extensive herbicide selection pressure, many waterhemp populations across the USA evolved resistance to multiple herbicides, leading to limited options for chemical control. Genome editing tools, including CRISPR/Cas9, have great potential in weed science, especially when coupled with a gene drive system for a sustainable weed management. The application of modern biotechnological tools requires availability of an efficient regeneration and genetic transformation system. The objectives of this research were to develop an efficient protocol for successful regeneration of waterhemp via callus culture and optimize an Agrobacterium-mediated transformation method for waterhemp. Hypocotyl explants cultured on callus induction medium (CIM), C6 containing 1.0 mg L−1 kinetin and 4.0 mg L−1 2,4-D for 2 wk followed by subculture into shoot induction medium (SIM), S8 containing 3.0 mg L−1 BAP, 0.05 mg L−1 2,4-D, and 0.3 mg L−1 GA3 for 4 to 6 wk was successful in shoot regeneration. Leaf explants failed to respond to any treatment combinations tested for regeneration. Using the hypocotyl explant–based regeneration protocol optimized in this research, Agrobacterium-mediated transformation was performed using uidA and hpt genes as a proof of concept. Although transgenic plants were not regenerated, molecular characterization of the transformed calluses confirmed the presence of transgenic genes, for example uidA and hpt. This is the first report of the tissue-cultured-based regeneration and Agrobacterium-mediated transformation of hypocotyl derived calluses in waterhemp, which is a step forward in exploring the possibilities of genome editing research in weed science. [ABSTRACT FROM AUTHOR]

Potato (Solanum tuberosum L.) is the third most important crop after rice and wheat in the world and it is one of the most efficient food crops producing more yield, alimental fibre, high-quality protein, minerals and vitamins than wheat, maize and rice per unit area and time and is recognized as a steady and healthy food crop. The production of seed potatoes and its supply chain face special problems because they are big in size, multiply slowly and large quantities are needed to plant the next crop. There is a huge gap between production and supply of quality planting materials of potato. So, it is important to develop a systematic protocol for production of planting materials at large scale. Given such conditions, the conception of an effective and reproducible in vitro regeneration system in popular potato cultivar(s) is crucial to produce high-quality planting material at large scale. Keeping this in mind, to produce genuine propagative material of high phytosanitary, physiological and genetic quality, effective and reproducible in vitro regeneration protocols of potato are indispensable. To attain such goals, potato sprouts were employed as explants sources and inoculated on MS medium fortified with different auxins and cytokinins in fluctuating proportions or alone as well as in diverse concentrations and amalgamations in combination with 30.0 g l−1 sucrose and 7.5 g l−1 agar. Induction medium MS3NB (MS + 3.0 mg l−1 NAA + 1.0 mg l−1 BA) was more responsive for callus induction (83.00%). Whilst nutrient media MS3BN (MS + 3.0 mg l−1 BAP + 1.0 mg l−1 NAA) displayed better in vitro competence, i.e. number(s) of shoot inducing explants (91.00%) and number(s) of shoot(s) per explant (6.50) along with shoot of bigger length (4.60). Whilst higher in vitro rooting response, viz., root proliferating efficiency (84.0%), number(s) of roots (9.0) with higher length (5.1) was exhibited by rooting medium MSIB (MS + 1.0 mg l−1 IBA). For inducing rooting in vitro, a reduced level of sucrose, i.e. 15.0 g l−l, was used. The regenerants were shifted to pots and acclimatized in polyhouse/net house during preliminary ablactating phase. Regenerants showed normal growth and morphology and were found efficacious in the external environment after hardening. Culture medium with higher regeneration aptitude obtained in present investigation may be used for mass in vitro propagation, micro tuber production and different biotechnological works including tailoring transgenic plants resistant/tolerant against diverse biotic and abiotic stresses. [ABSTRACT FROM AUTHOR]

The initiation of cultivars for hazelnut micropropagation from mature field-grown plants is challenging due to high contamination levels and low regeneration rates. This study aimed to develop an efficient protocol for establishing in vitro cultures, the crucial first step in hazelnut micropropagation. Three independent experiments were conducted to achieve this goal. In the first experiment, plant materials were collected directly from the field at different times of the year to initiate cultures, with July to August identified as the most suitable period. In the second experiment, one-year-old potted suckers underwent a sanitation program in the greenhouse and were subjected to thermotherapy in the growth chamber before being utilized as explant sources. Nodal explants obtained from these plants were then used to initiate cultures. In the third experiment, various doses and durations of sterilization agents were applied to optimize surface sterilization. The combination of 1% silver nitrate (AgNO3), 0.1% mercury chloride (HgCl2), and 3% hydrogen peroxide (H2O2) after treatments with 70% ethanol and 2% sodium hypochlorite (NaOCl) yielded the best results, with the highest percentage of viable shoots. Overall, thermotherapy treatment to the mother plant was the most effective method for initiating cultures from plants in highly contaminated environments. While cultures initiated with explants from field-grown plants exhibited 53.7% visually clean shoots, this rate increased to 97.4% in plants subjected to thermotherapy. This study, for the first time, evaluated the effectiveness of applying thermotherapy to mother plants to reduce the incidence of microbial contaminants in the establishment stage of hazelnut in vitro propagation.Key message: The present study revealed that applying thermotherapy to mother plants effectively reduces microbial contamination during in vitro culture establishment and enhances overall culture success. [ABSTRACT FROM AUTHOR]

Determining foliar mineral status of tissue cultured shoots can be costly and time consuming, yet hyperspectral signatures might be useful for determining mineral contents of these shoots. In this study, hyperspectral signatures were acquired from tissue cultured little-leaf mockorange (Philadelphus microphillus) shoots to determine the feasibility of using this technology to predict foliar nitrogen and calcium contents. After using a spectroradiometer to take hyperspectral images for determining foliar N and Ca contents, the correlation between the hyperspectral bands, vegetation indices, and hyperspectral features were calculated from the spectra. Features with high correlations were selected to develop the models via different regression methods including linear, random forest (RF), and support vector machines. The results showed that non-linear regression models developed through machine learning techniques, including RF methods and support vector machines provided satisfactory prediction models with high R2 values (%N by RF with R2 = 0.72, and %Ca by RF with R2 = 0.99), that can estimate nitrogen and calcium content of little-leaf mockorange shoots grown in vitro. Overall, the RF regression method provided the most accurate and satisfactory models for both foliar N and Ca estimation of little-leaf mockorange shoots grown in tissue culture.Key message: Hyperspectral signatures were used to estimate foliar %N and %Ca of micropropagated Philadelphus microphyllus (little-leaf mockorange) shoots. Non-linear regression models provided satisfactory prediction with high R2 values. [ABSTRACT FROM AUTHOR]