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Musculoskeletal injuries, including tendinopathies, present a significant clinical burden for aging populations. While the biological drivers of age‐related declines in tendon function are poorly understood, it is well accepted that dysregulation of extracellular matrix (ECM) remodeling plays a role in chronic tendon degeneration. Senescent cells, which have been associated with multiple degenerative pathologies in musculoskeletal tissues, secrete a highly pro‐inflammatory senescence‐associated secretory phenotype (SASP) that has potential to promote ECM breakdown. However, the role of senescent cells in the dysregulation of tendon ECM homeostasis is largely unknown. To assess this directly, we developed an in vitro model of induced cellular senescence in murine tendon explants. This novel technique enables us to study the isolated interactions of senescent cells and their native ECM without interference from age‐related systemic changes. We document multiple biomarkers of cellular senescence in induced tendon explants including cell cycle arrest, apoptosis resistance, and sustained inflammatory responses. We then utilize this in vitro senescence model to compare the ECM remodeling response of young, naturally aged, and induced‐senescent tendons to an altered mechanical stimulus. We found that both senescence and aging independently led to alterations in ECM‐related gene expression, reductions in protein synthesis, and tissue compositional changes. Furthermore, MMP activity was sustained, thus shifting the remodeling balance of aged and induced‐senescent tissues towards degradation over production. Together, this demonstrates that cellular senescence plays a role in the altered mechano‐response of aged tendons and likely contributes to poor clinical outcomes in aging populations. [ABSTRACT FROM AUTHOR]

Background Objectives Materials and Methods Results Conclusions and Clinical Relevance Canine superficial pyoderma is a common bacterial skin infection of dogs, generally caused by Staphylococcus pseudintermedius. The C4 strain of Staphylococcus felis was recently discovered to have strong antimicrobial activity against S. pseudintermedius in mice.We aimed to evaluate in vitro if this antimicrobial activity was maintained using a novel canine skin explant model.Punch biopsies (8 mm) of skin from recently euthanised dogs were collected and placed into six‐well plates on top of an agarose pedestal.Histological examination of the skin explants showed an intact dermal–epidermal organisation and a stratum corneum that was successfully colonised by S. pseudintermedius after topical application. The number of colony forming units of S. pseudintermedius showed a 2 log increase after 24 h colonisation, indicating that the explant supported bacterial growth. By contrast, co‐treatment with S. felis C4 live bacteria and its sterile protein product significantly reduced the growth of a methicillin‐susceptible (ST540, p = 0.0357) and a methicillin‐resistant (MR) strain (ST71, p = 0.0143) of S. pseudintermedius. No detectable bacteria were recovered from or visualised on skin 24 h posttreatment with the S. felis C4 sterile protein product.Using a novel canine explant model, we demonstrate that the S. felis C4 strain inhibits the growth of S. pseudintermedius and that it is a promising candidate for a new probiotic therapy to treat cutaneous infections caused by S. pseudintermedius, including MR strains. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Henan Agricultural Sciences is the property of Editorial Board of Journal of Henan Agricultural Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

To establish an efficient regeneration method for the rare and endangered plant Mussaenda anomala to address problems regarding its reproductive obstacles and scarce populations. In this study, the terminal buds, axillary buds, stem segments with two axillary buds, stem segments with two axillary buds and one terminal bud, and leaves of M. anomala were used as explants. The effects of different explants and disinfection methods, plant growth regulators and substrates on plant regeneration were explored. The following results were obtained: (1) The terminal bud was a suitable explant for M. anomala tissue culture, and the disinfection method utilized was treatment with 0.2% HgCl2 for 8 min. (2) Initiate medium: MS basic medium supplemented with 0.5 mg/L 6-BA and 0.2 mg/L IBA for the high germination rate (100%) and the maximum bud height (1.70 cm) of the terminal bud. (3) Proliferation medium: MS basic medium supplemented with 3.0 mg/L 6-BA and 0.2 mg/L IBA for a high proliferation rate (96%) and proliferation time (6.0) of terminal buds. (4) Proliferation medium supplemented with 0.7 mg/L GA3 significantly increased the bud heights of multiple buds. (5) Rooting medium: MS basic medium supplemented with 0.5 mg/L IBA and 0.5 mg/L IAA for a high rooting rate (88%), root number (12.0) and root length (5.07 cm). (6) The optimal substrate for seedling acclimation and transplanting was perlite: vermiculite (1:1), which resulted in the highest survival rate (97%) and plant height (5.89 cm), as well as better growth potential for seedlings. The surfaces of M. anomala explants are densely covered with trichome, which increased the difficulty of disinfection; the plant growth regulators directly affected the growth and development in the regeneration process of M. anomala, and the substrate significantly affected the survival rate and height growth for seedling acclimation. [ABSTRACT FROM AUTHOR]

Left ventricular assist devices (LVADs) are excellent therapies for advanced heart failure patients either bridged to transplant or for lifetime use. LVADs also allow for reverse remodeling of the failing heart that is often associated with functional improvement. Indeed, growing enthusiasm exists to better understand this population of patients, whereby the LVAD is used as an adjunct to mediate myocardial recovery. When patients achieve benchmarks suggesting that they no longer need LVAD support, questions related to the discontinuation of LVAD therapy become front and center. The purpose of this review is to provide a surgical perspective on the practical and technical issues surrounding LVAD deactivation. [ABSTRACT FROM AUTHOR]

Aims The extravascular implantable cardioverter-defibrillator (EV ICD) has been shown to be safe and effective for patients at risk of sudden cardiac death, but little is known about EV ICD lead removal in humans. This analysis aimed to characterize the EV ICD lead removal experience thus far. Methods and results This was a retrospective analysis of lead removals from the EV ICD Pilot, Pivotal, and Continued Access Studies. Patients with a successful EV ICD implant who underwent lead removal were included. The main objective was lead removal success. Ancillary objectives included characterizing technique used, procedure complications, and reimplantation status. An EV ICD system was successfully implanted in 347 patients across the 3 studies (25.9% female; 53.4 ± 13.3 years; left ventricular ejection fraction: 39.7 ± 15.9). Of these patients, 29 (8.4%) underwent lead removal with a mean lead dwell time of 12.6 ± 14.3 months (0.2–58.4). The main reason for lead removal was lead dislodgement (n = 9, 31.0%). Lead removal was successful in 27/29 (93.1%) cases [100% (19/19) success rate <1 year and 80% (8/10) success rate >1 year post-implant]. Simple traction was used in 22/26 (84.6%) and extraction tools in 4/26 (15.4%) successful cases where technique was known. No complications were reported for any of the removal procedures. All 11 EV ICD reimplant attempts were successful. Conclusion Complete removal of the EV ICD lead was successful in 93.1% of cases, and simple traction was sufficient in most instances. Based on these results, lead removal from the substernal space was safe and achievable up to 3 years post-implant. [ABSTRACT FROM AUTHOR]

Chrysanthemum morifolium '11-C-2' is a variety of chrysanthemums with high ornamental and tea value, experiencing significant market demand. However, as cultivation areas expand, issues such as viral infection, germplasm degradation, low proliferation coefficient, and slow proliferation rate arise, necessitating the establishment of an efficient in vitro regeneration system. This study, based on the principles of orthogonal experimental design, explored the regeneration system of Chrysanthemum cultivar '11-C-2' using sterile seedlings. The research focused on three key stages: adventitious bud differentiation, rooting culture, and acclimatization–transplantation, employing shoot-bearing stem segments and leaves as explants. The findings indicate that the optimal explant for the Chrysanthemum '11-C-2' sterile seedlings is the shoot-bearing stem segment. The best medium for adventitious bud differentiation was determined to be MS supplemented with 1.5 mg/L 6-BA and 0.5 mg/L NAA. Bud differentiation began on day 17 with a 100% differentiation rate, completing around day 48. The maximum differentiation coefficient reached 87, with an average of 26.67. The adventitious buds were then cultured for rooting in the optimal medium of 1/2 MS supplemented with 0.1 mg/L NAA. Rooting was initiated on day 4 and was completed by day 14, achieving a rooting rate of 97.62%. After a 5-day acclimatization under natural light, the rooted seedlings were transplanted into a growth substrate with a peat-to-vermiculite ratio of 1:2. The plants exhibited optimal growth, with a transplantation survival rate of 100%. The findings provide data support for the efficient large-scale propagation of '11-C-2' and lay the foundation for germplasm preservation and genetic transformation research of tea chrysanthemums. [ABSTRACT FROM AUTHOR]

The purpose of the study is to optimize the trophic and hormonal determination of raspberry reproduction in vitro and post-aseptic adaptation ex vitro. The research was carried out on the remontant raspberry variety Delniwa of Polish selection. Laboratory studies were carried out in the conditions of the Tevitta TM laboratory (Ltd "Agrofirm Blagodatne," Cherkasy region), and experiments were laid in four stages: introduction into aseptic culture, multiplication, induction of rhizogenesis, and post-aseptic adaptation, including photoautotrophic grafting. A medium for obtaining an aseptic culture and a media for multiplication have been selected. The rate of regeneration of explants of different origins, and the percentage of regenerants from them, was compared. The dynamics of the influence of cytokinin concentrations during long-term reproduction in vitro were revealed. Cultivation on a medium with a high content of benzylaminopurine and kinetin showed the effect of accumulation of phytotoxicity. The expediency of growing graft donors for grafting at low kinetin concentrations to improve the induction of rhizogenesis is substantiated. Regenerants from media with a predominance of auxins over cytokinins had a higher regeneration potential. According to indicators of suitability for survival, perlite substrates prevailed over peat ones; however, regenerants of a smaller height were formed on them. The symptoms and harmful effects of excessive watering on plants have been established. A biomethod of insect control using insectivorous plants of the genus Drosera has been developed. The rate of rhizogenesis in bioreactor conditions with humidified and carbon dioxide-enriched air significantly exceeded root formation in conventional moist chambers. The duration of growing an adapted and well-rooted seedling in a bioreactor was 21 days, compared to 30 or more days in the control version of the experiment. The plants did not lose turgor when planted in closed soil. Adapted regenerants are successfully used as a planting material and can be used in photoautotrophic microclonal propagation as donors of cuttings. Under such conditions, the regeneration efficiency is maintained for five cutting generations. [ABSTRACT FROM AUTHOR]

One of the main problems of fruit cultivation, especially peaches and almonds, is the lack of the application of the existing clone rootstocks such as the promising hybrid of peaches and almonds and its unavailability due to its propagation in Iran. So the seedling rootstocks are inevitably used. However these rootstocks cause many problems for the growers due to the lack of uniformity. In order to solve this problem, the effect of different concentrations of indole butyric acid on the rooting of hardwood cuttings ‘TF92’ (promising hybrid of peach and almond) was tested in a private sector greenhouse in Karaj equipped with a heating pad during 2019. In this paper, the effect of 4 different levels of indole butyric acid hormone (IBA) 0, 1500, 2500 and 3500 parts per million in solution on hardwood cuttings was studied in a completely randomized design in three replications. The results showed that there was a significant difference between various concentrations of indole butyric acid in terms of the effect on the characteristics of the mode of investigation such as the percentage of rooting, the number of roots formed, the dry weight of the roots, the length of the roots and the length of the resulting branches in the cuttings. Thus the highest rooting percentage (78.45) was found in the indole butyric acid (IBA) hormone treatment with a concentration of 2500 mg L-1 compared to other treatments. Therefore, this method can be recommended as a suitable treatment for the rooting of wood cuttings of this promising emerging rootstock. Also, the highest percentage of rooted plantlets was obtained on Murashige and Skoog medium (MS) supplemented with 1 mg L-1 IBA. According to this study, the concentrations of 1 mg L-1 IBA results in the maximum rooting, and it has been suggested as a suitable hormone for in vitro rooting of ‘TF92’.

Drought stress can impair plant growth and development. Nitric oxide (NO), a gas signaling molecule, delays injuries caused by abiotic stress. This study explored the roles of NO during adventitious rooting in cucumber (Cucumis sativus L.) under drought stress. Our results found that 0.05% (W/V) polyethylene glycol 6000 (PEG) treatment significantly suppressed adventitious root formation, but 10 μM GSNO (a NO donor) treatment could significantly reverse the negative effect of drought stress. Under drought stress, MDA, H2O2 and O2·− accumulation were increased during rooting. Nevertheless, the application of exogenous NO decreased reactive oxygen (ROS) accumulation, revealing that NO might play an essential role in improving drought tolerance. Compared with PEG treatment, PEG + GSNO treatment enhanced the level of glutathione (GSH), ascorbic acid (AsA), and GSH/GSSG ratio, while decreasing glutathione disulfde (GSSG) content. In comparison with PEG treatment, PEG + GSNO treatment significantly increased the activity of antioxidant enzymes (APX, AAO, GR, and MDHAR) and the expression level of related enzyme genes (CsAPX, CsAAO, CsGR, and CsMDHAR). In addition, PEG + GSNO treatment also increased endogenous NO and S-nitrosothiol (SNO) content, nitrate reductase (NR) activity, and total S-nitrosylated proteins, and the expression of CsNR, while PEG + GSNO treatment decreased S-nitrosoglutathione reductase (GSNOR) activity, suggesting that protein S-nitrosylation might be involved in adventitious rooting under drought stress. In summary, our results indicate that NO could promote adventitious root formation under drought stress via regulating ascorbate glutathione (AsA-GSH) pathway cycle and protein posttranslational S-nitrosylation. [ABSTRACT FROM AUTHOR]

Deafness-causing deficiencies in otoferlin (OTOF) have been addressed preclinically using dual adeno-associated virus (AAV)-based approaches. However, timing of transduction, recombination of mRNA, and protein expression with dual hybrid AAV methods methods have not previously been characterized. Here, we have established an ex vivo assay to determine the kinetics of dual-AAV mediated expression of OTOF in hair cells of the mouse utricle. We utilized two different recombinant vectors that comprise DB-OTO, one containing the 5′ portion of OTOF under the control of the hair cellspecific Myo15 promoter, and the other the 3′ portion of OTOF. We explored specificity of the Myo15 promoter in hair cells of the mouse utricle, established dose response characteristics of DB-OTO ex vivo in an OTOF-deficient mouse model, and demonstrated tolerability of AAV1 in utricular hair cells. Furthermore, we established deviations from a one-to-one ratio of 5′ to 3′ vectors with little impact on recombined OTOF. Finally, we established a plateau in quantity of recombined OTOF mRNA and protein expression by 14 to 21  days ex vivo with comparable recovery timing to that in vivo model. These findings demonstrate the utility of an ex vivo model system for exploring expression kinetics and establish in vivo and ex vivo recovery timing of dual AAV-mediated OTOF expression. [ABSTRACT FROM AUTHOR]

Traction force that can be applied to an extraction rail is based on lead tensile strength, a product of its construction. A strong rail allows safe advancement of the extraction sheath. This study expands previous work providing strategies to optimize INGEVITY rail strength. The purpose of this study was to measure forces that leads encounter in a simulated extraction procedure, determine lead response, and develop extraction recommendations for INGEVITY, INGEVITY+, and FINELINE II lead families. Leads were positioned in a simulated right atrial appendage implant. Subsequent traction forces enabled evaluation of lead tensile strength and effectiveness of preparation/extraction techniques. Significant findings include (1) preserving the lead terminal pin did not decrease lead tensile strength and typically maximized it; (2) the weakest region is between the cathode and anode; (3) mid lead scar increases traction force tolerance until that scar is removed; and (4) optimal rail strength was observed using a multivenous approach with a femoral snare. Unique lead family findings include increased tensile strength of FINELINE II polyurethane vs silicone and INGEVITY active fixation vs passive fixation. This study teaches the implanting clinician there are specific extraction techniques available to improve the removal of leads that may be the best option for a patient's clinical needs. Bench testing demonstrates that lead construction drives lead behavior during an extraction. Preserving the lead terminal pin provides consistent and, in most cases, optimal rail strength. If clinically indicated, a multivenous approach using a femoral snare significantly increases rail strength and protects the vulnerable lead tip. [ABSTRACT FROM AUTHOR]

The goal of the research was to develop a technology for introducing Salicornia europaea L.into in vitro culture. Methods of cultivating meristems and callus cultures were studied. To cultivate meristems, the tip of the apical bud were used as an explant. Fragments of stems and leaves were used toobtain callus tissue. To study the influence of various factors on germination, seeds were soaked in sterile tap water and in solutions of gibberellin, cytokinin, auxin and NaCl, and were also subjected to cold stratification (independently and with subsequent placement in Knop’s agarized medium). Salicornia seeds were sterilized with various antiseptics: 70% alcohol, 10% aqueous solution of sodium hypochlorite («Belizna»), amoxicillinand 3% hydrogen peroxide. The ability of the culture to form callus was studied in MS medium. As a result, it was determined that the highest germination of seeds was observed in Knop medium after treating theseeds with a suspension of green algae of the Scenedesmus genus, as well as after preliminary cold stratification, and slightly less after treating the seeds with a solution of NaCl and gibberellic acid. The most effective method of seed sterilization turned out to be treatment with alcohol followed by treatment with sodium hypochlorite. A comparative analysis of seed germination in filter paper in Petri dishes, Knop agar medium, Murashige and Skoog (hormone-free) was carried out. The ability of S. europaea L. to form callusin MS medium with phytohormones was assessed. Conclusion. To improve germination, it is recommended to subject the seeds to cold stratification. To obtain aseptic explants quickly, it is recommended to germinate the seeds in the Knop nutrient medium, having previously sterilized them with alcohol, then with sodiumhypochlorite, followed by washing with distilled water. The most suitable type of microclonal propagationfor S. europaea L. is the production of callus tissue followed by induction of organogenesis or embryogenesis.

Genomic-based precision medicine has not only improved tumour therapy but has also shown its weaknesses. Genomic profiling and mutation analysis have identified alterations that play a major role in sarcoma pathogenesis and evolution. However, they have not been sufficient in predicting tumour vulnerability and advancing treatment. The relative rarity of sarcomas and the genetic heterogeneity between subtypes also stand in the way of gaining statistically significant results from clinical trials. Personalized three-dimensional tumour models that reflect the specific histologic subtype are emerging as functional assays to test anticancer drugs, complementing genomic screening. Here, we provide an overview of current target therapy for sarcomas and discuss functional assays based on 3D models that, by recapitulating the molecular pathways and tumour microenvironment, may predict patient response to treatments. This approach opens new avenues to improve precision medicine when genomic and pathway alterations are not sufficient to guide the choice of the most promising treatment. Furthermore, we discuss the aspects of the 3D culture assays that need to be improved, such as the standardisation of growth conditions and the definition of in vitro responses that can be used as a cut-off for clinical implementation.

3D human tissue models/microphysiological systems (3D/MPS) can represent human physiology. Linking organ models can improve biological fidelity and predictive capability. 3D/MPS may be infected with HIV, enabling studies of pathogenesis, host–virus interactions, inflammation, neuropathogenesis, and HIV reservoirs. Integrating immune cells or lymphoid tissues into 3D/MPS facilitates study of HIV immune responses, leukocyte depletion, immune ontogeny, immunotherapies, and vaccines. Application of 3D/MPS to therapeutic development facilitates sophisticated preclinical pharmacology, in silico modeling, predictions of human response and safety, and evaluation of potency of biologics. Tissue sources and/or modifications permit study of unique conditions, including TB co-infection, substance use, and special populations (e.g., medically fragile, pregnant or lactating people, and children). Three-dimensional (3D) human tissue models/microphysiological systems (e.g., organs-on-chips, organoids, and tissue explants) model HIV and related comorbidities and have potential to address critical questions, including characterization of viral reservoirs, insufficient innate and adaptive immune responses, biomarker discovery and evaluation, medical complexity with comorbidities (e.g., tuberculosis and SARS-CoV-2), and protection and transmission during pregnancy and birth. Composed of multiple primary or stem cell-derived cell types organized in a dedicated 3D space, these systems hold unique promise for better reproducing human physiology, advancing therapeutic development, and bridging the human–animal model translational gap. Here, we discuss the promises and achievements with 3D human tissue models in HIV and comorbidity research, along with remaining barriers with respect to cell biology, virology, immunology, and regulatory issues. [ABSTRACT FROM AUTHOR]

Hippophae gyantsensis Lian is a pioneering tree species in Chinese forestry ecological engineering, known for its robust stress tolerance, water retention capacity, and soil improvement qualities. However, the lack of rapid nursery technology has been a significant impediment to the development of the H. gyantsensis industry. In the present study, we have successfully established a tissue culture regeneration system for H. gyantsensis. The most effective methods for seed disinfection, ensuring sterility in seedlings, were found to be 75% alcohol disinfection for 40 s and 10% sodium hypochlorite disinfection for 10 min. The best media tested for callus induction in cotyledons and hypocotyls of sterile seedlings were 1/3 MS + 0.5 mg/L KT + 0.75 mg/L NAA and 1/3 MS + 0.3 mg/L 6-BA + 1.5 mg/L IBA, respectively. As the explants, cotyledons yielded larger calli with a greater size and differentiation ability than hypocotyls. For the induction of indeterminate shoots and proliferation, the most suitable media were 1/3 MS + 0.5 mg/L IAA + 0.75 mg/L 6-BA and 1/3 MS + 1.0 mg/L 6-BA + 0.05 mg/L IBA + 0.1 mg/L KT, respectively. Lastly, the best worked rooting formulation was 1/4 MS + 0.3 mg/L IBA. This study marks a significant milestone in the establishment of a systematic tissue culture regeneration system for H. gyantsensis, which will facilitate the industrial rapid propagation of high-quality seedlings and provide the foundation for improvement through genetic transformation. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Northwest A & F University - Natural Science Edition is the property of Editorial Department of Journal of Northwest A&F University (Natural Science Edition) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Spinal cord stimulation (SCS) is a therapeutic modality for the treatment of various chronic pain conditions that has rapidly evolved over the past 50 years. Unfortunately, over time, patients implanted with SCS undergo a habituation phenomenon leading to decreased pain relief. Consequently, the discovery of new stimulation waveforms and SCS applications has been shown to prolong efficacy and reduce explantation rates. This article explores various SCS waveforms, their applications, and proposes a graded approach to habituation mitigation. We suspect the neural habituation phenomenon parallels that seen in pharmacology. Consequently, we urge further exploration of the early introduction of these stimulation strategies to abate spinal cord stimulation habituation. [ABSTRACT FROM AUTHOR]

Copyright of Eurasian Journal of Applied Biotechnology is the property of National Center for Biotechnology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

This research aimed at in vitro propagations of bananas originating from India. Banana (Musa spp.) is a nutritious fruit but shows susceptibility to specific diseases. A traditional method of its propagation is through the separation of suckers, although it may culminate in the transmission of nematodes, parasitic organisms, and viral diseases. In the past two decades, plant tissue culture techniques have facilitated the production of disease-free plantlets. Tissue culturing bananas can involve different explants, including shoot tips, suckers, leaves, and flower buds. Each responds differently to the presence and absence of light. In the current research, explant cultures were placed in light or dark incubation for identical durations, i.e., one to six months, to monitor their growth and development. The color intensity of the explants changed in response to different photoperiods. Young flower buds, mature flower buds, and suckers developed a higher color intensity when placed in light than in dark conditions. However, the opposite occurred in leaf explants that grew optimally in the dark. The results indicated that banana growth occurred productively from sucker explants in different light conditions and variable durations. The results were optimal when employing sucker explants, which exhibited the fastest growth. [ABSTRACT FROM AUTHOR]

Since their introduction in the 1960's, the safety of silicone breast implants has remained contentious due to concerns regarding carcinogenicity as well as a growing array of adverse psychiatric symptoms, which have now been termed 'Breast Implant Illness.' This article aims to explore the merits of a psychiatric approach to treating Breast Implant Illness by outlining how it is defined by psychiatric symptoms and categorized alongside other psychiatric illnesses. Furthermore, it is unclear whether the pathology of Breast Implant Illness is purely medical or psychiatric. However, the efficacy of the medical approach to treatment through a process called explantation, which involves removal of the implant and surrounding scar tissue, or capsule, is not strongly supported by existing data. A psychiatric approach to treatment, in conjunction with explanation, thus holds potential in remedying the novel and poorly understood Breast Implant Illness. Level of Evidence V This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266. [ABSTRACT FROM AUTHOR]

The octoploid-cultivated strawberry variety Benihope (Fragaria × ananassa Duch cv. Benihope) is an important commercial plant. It is highly susceptible to different diseases, which ultimately leads to a reduction in yield. Gene-editing methods, such as CRISPR/Cas9, demonstrate potential for improving disease resistance in the strawberry cv. Benihope. Establishing a plant regeneration system suitable for CRISPR/Cas9 gene editing is crucial for obtaining transgenic plants on a large scale. This research established a callus induction and plant regeneration system for Agrobacterium-mediated CRISPR/Cas9 gene editing in strawberry cv. Benihope by evaluating multiple types of explants and various plant growth regulators throughout the entire tissue culture process. The results showed that the efficiency of callus induction is strongly influenced by the type of explant and is highly sensitive to the combination of plant growth regulators. Among the different plant growth regulators employed, thidiazuron (TDZ), in combination with 2,4-dichlorophenoxyacetic acid (2,4-D), effectively induced callus formation and plant regeneration from explants derived from nutrient tissues such as runner tips and crowns. In addition, the regeneration experiment demonstrated that the addition of polyvinylpyrrolidone (PVPP) to the shoot regeneration medium could inhibit tissue browning. The gene-edited plants in which some or all of the Fvb7-1, Fvb7-2, Fvb7-3, and Fvb7-4 genes in the MLO (Mildew resistance Locus O) gene family were knocked out by CRISPR/Cas9 system were obtained by applying the plant regeneration system developed in this study. [ABSTRACT FROM AUTHOR]

Deafness-causing deficiencies in otoferlin (OTOF) have been addressed preclinically using dual adeno-associated virus (AAV)-based approaches. However, timing of transduction, recombination of mRNA, and protein expression with dual hybrid AAV methods methods have not previously been characterized. Here, we have established an ex vivo assay to determine the kinetics of dual-AAV mediated expression of OTOF in hair cells of the mouse utricle. We utilized two different recombinant vectors that comprise DB-OTO, one containing the 5′ portion of OTOF under the control of the hair cell-specific Myo15 promoter, and the other the 3′ portion of OTOF. We explored specificity of the Myo15 promoter in hair cells of the mouse utricle, established dose response characteristics of DB-OTO ex vivo in an OTOF-deficient mouse model, and demonstrated tolerability of AAV1 in utricular hair cells. Furthermore, we established deviations from a one-to-one ratio of 5′ to 3′ vectors with little impact on recombined OTOF. Finally, we established a plateau in quantity of recombined OTOF mRNA and protein expression by 14 to 21 days ex vivo with comparable recovery timing to that in vivo model. These findings demonstrate the utility of an ex vivo model system for exploring expression kinetics and establish in vivo and ex vivo recovery timing of dual AAV-mediated OTOF expression.

Abstract Background Traditional breeding methods have long been employed worldwide for the evaluation and development of pepper cultivars. However, these methods necessitate multiple generations of screening, line development, evaluation, recognition, and crossing to obtain highly homozygous lines. In contrast, in vitro anther-derived microspore culture represents a rapid method to generate homozygous lines within a single generation. In the present study, we have optimized a protocol for microspore embryogenesis from anther cultures of pepper hybrids Orobelle and Bomby. Results We achieved early and successful embryo formation from both genotypes by subjecting the buds to a cold pretreatment at 4 °C for 4 days. Our optimized culture medium, comprised of MS medium supplemented with 4 mg/L NAA, 1 mg/L BAP, 0.25% activated charcoal, 2.6 g/L gelrite, 30 g/L sucrose, and 15 mg/L silver nitrate, exhibited the highest efficiency in embryo formation (1.85% and 1.46%) for Orobelle and Bomby, respectively. Furthermore, successful plant regeneration from the anther derived microspore embryos was accomplished using half-strength MS medium fortified with 2% sucrose and 0.1 mg/L 6-benzylaminopurine (BA), solidified with 2.6 g/L gelrite. The ploidy status of the microspore-derived plantlets was analyzed using flow cytometry technique. Notably, the haploid plants exhibited distinct characteristics such as reduced plant height, leaf length, leaf width, and shorter internode length when compared to their diploid counterparts derived from seeds. Conclusion Our findings highlight the potential of anther culture and microspore embryogenesis as an advanced method for accelerating pepper breeding programs, enabling the rapid production of superior homozygous lines.

Barangan banana (Musa acuminata L.) is one of the superior types of banana in the West Tanjung Jabung district, Jambi Province - Indonesia. Barangan bananas are very popular with the public and have high economic value. The problem faced in conventional cultivation and development of Barangan bananas is that it is difficult to obtain quality banana seeds in large and uniform quantities, so to overcome this we need alternative technology for propagating banana seeds through modern techniques such as plant tissue culture techniques. The advantage of this technique is that it is able to produce plant seeds en masse uniformly and in a relatively short time. In addition, the resulting progeny is also conditioned to be free from pests and diseases, especially diseases caused by bacteria and fungi.

Exploring alternative avenues, in vitro culture emerges as a promising option for potential bioactive compound sources. However, compared to intact plants, only a few cultures demonstrate efficient synthesis of secondary metabolites. Elicitors have gained prominence as stress agents for enhancing in vitro micropropagation in specific tissues, organs, and cells. Recent advancements in plant tissue culture involve elicitors, opening new possibilities for in vitro production of crucial food crops. This research aimed to investigate the impact of three elicitors (Activane®, Micobiol®, and Stemicol®) on germination and in vitro multiplication of two tomato cultivars explants, employing both direct and indirect in vitro organogenesis. Among the texted elicitors, Micobiol® emerged as a successful elicitor, promoting optimal seed germination, survival, and 100% growth compared to the 80% in the control group. Further, Activane® exhibited a favourable induction response and achieved 96%, 95%, and 100% in weight and diameter of callus, yet various elicitor concentrations did not exert significant influence across treatments. In conclusion, an effective disinfection and in vitro implantation of tomato seeds ensured successful germination, promoting seedling survival and growth. Various elicitors positively impacted in vitro organogenesis, particularly in root induction, with higher survival percentages in acclimatized plants. The study guides future research on elicitor treatments for largescale tomato in vitro propagation, emphasizing the need to identify optimal elicitor concentrations. [ABSTRACT FROM AUTHOR]

Background: Traditional breeding methods have long been employed worldwide for the evaluation and development of pepper cultivars. However, these methods necessitate multiple generations of screening, line development, evaluation, recognition, and crossing to obtain highly homozygous lines. In contrast, in vitro anther-derived microspore culture represents a rapid method to generate homozygous lines within a single generation. In the present study, we have optimized a protocol for microspore embryogenesis from anther cultures of pepper hybrids Orobelle and Bomby. Results: We achieved early and successful embryo formation from both genotypes by subjecting the buds to a cold pretreatment at 4 °C for 4 days. Our optimized culture medium, comprised of MS medium supplemented with 4 mg/L NAA, 1 mg/L BAP, 0.25% activated charcoal, 2.6 g/L gelrite, 30 g/L sucrose, and 15 mg/L silver nitrate, exhibited the highest efficiency in embryo formation (1.85% and 1.46%) for Orobelle and Bomby, respectively. Furthermore, successful plant regeneration from the anther derived microspore embryos was accomplished using half-strength MS medium fortified with 2% sucrose and 0.1 mg/L 6-benzylaminopurine (BA), solidified with 2.6 g/L gelrite. The ploidy status of the microspore-derived plantlets was analyzed using flow cytometry technique. Notably, the haploid plants exhibited distinct characteristics such as reduced plant height, leaf length, leaf width, and shorter internode length when compared to their diploid counterparts derived from seeds. Conclusion: Our findings highlight the potential of anther culture and microspore embryogenesis as an advanced method for accelerating pepper breeding programs, enabling the rapid production of superior homozygous lines. [ABSTRACT FROM AUTHOR]

In this study; the development of an effective surface sterilization for Kozdere, Zengibar and Chandler walnut cultivars and the interaction of different media and 6-benzylaminopurine (BAP) concentrations in culture initiation were investigated. Axillary buds of walnut cultivars were used as explants. The first stage of tissue culture studies is surface sterilization. Plants with successful surface sterilization can be micropropagated. In this study, walnut materials were surface sterilized by using sodium hypochlorite at doses of 20, 30 and 40% and copper sulphate in addition to sodium hypochlorite. The least contamination (8.16%) and darkening (22.45%) and the highest bud set (69.39%) were observed in Kozdere cultivar treated with 30% sodium hypochlorite and copper sulphate. Murashige and Skoog (MS) and Driver and Kuniyuki Walnut (DKW) media were used as culture initiation media. 0.5, 1 and 2 mg L-1 BAP were added to the media. DKW medium was found more successful than MS medium in the propagation of walnut cultivars. The addition of 2 mg L-1 BAP to DKW medium gave the best results for shoot elongation, number of shoots per explant and number of leaves for all cultivars. Among the walnut cultivars, Kozdere cultivar was found statistically significantly different from Zengibar and Chandler cultivars in shoot elongation measurements. As a result, it was revealed that Kozdere cultivar can be used for in vitro propagation of walnut cultivars. It is seen that more comprehensive studies are needed to determine the method for propagation of Zengibar and Chandler cultivars in tissue culture. [ABSTRACT FROM AUTHOR]

The development of an efficient and consistent callus-mediated in vitro regeneration protocol is crucial for biotechnological approaches aimed at improving chickpea, an economically important crop legume. In this study, we assess the effectiveness of callus-mediated regeneration in different chickpea genotypes. Through in vitro screening of explants, we identified the Indian cultivar Pusa 240 as a favourable genotype with higher efficiency of somatic embryogenesis and in vitro plant regeneration. Building upon this finding, we have successfully established two distinct protocols for chickpea callus-mediated somatic embryogenesis, utilizing leaf and hypocotyl explants obtained from the Pusa 240 genotype. These protocols achieved plant regeneration efficiencies of 27% using leaf explants and 46.6 − 66% using hypocotyl explants. Extensive literature review and comparative analysis underscored the superiority of our current protocol. Subsequently, the regenerated plants were successfully acclimatized and transferred to the greenhouse, exhibiting normal phenotypic growth. This detailed regeneration method will provide a valuable resource for chickpea genetic transformations and the generation of large mutant populations where embryogenesis via callus formation is required. The protocol presented here establishes a powerful tool for studying the functional genomics of chickpea plants and lays the foundation for future advancements in this field. [ABSTRACT FROM AUTHOR]

Copyright of Anadolu (1300-0225) is the property of Anadolu Dergisi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Phocine distemper virus (PDV) is a significant cause of mortality for phocid seals; however, the susceptibility of otariids to this virus is poorly understood. The authors used a lymph-node explant culture system from California sea lions (Zalophus californianus, CSL) to investigate: (1) the role of signaling lymphocyte activation molecule (SLAM) and nectin-4 in PDV infection and their cellular expression patterns, (2) if PDV induces transcriptional regulation of cell-entry receptors, and (3) the involvement of apoptosis in PDV infection. PDV replicated in the lymph-node explants with peak replication 3 days post-infection (dpi), but the replication was not sustained 4 to 5 dpi. The PDV+ cells co-localized SLAM and nectin-4. These cells expressed IBA1, indicating a histiocytic lineage. Comparison of receptor expression between infected and mock-infected lymph nodes suggested transcriptional downregulation of both receptors during the initial stage of infection and upregulation during the late stage of infection, but the values lack of statistical significance. Cleaved caspase-3+ cells were slightly increased in the infected lymph nodes compared with the mock-infected lymph node from 1 to 4 dpi, but without statistical significance, and a few apoptotic cells co-expressed PDV. The results suggest that lymph-node explants might be an important model to study PDV pathogenesis. CSLs have the potential to be infected with PDV, as they express both cell-entry receptors in histiocytes. The lack of statistical significance in the PDV replication, transcriptional regulation of viral receptors, and changes in apoptosis suggest that although CSL might be infected by PDV, they might be less susceptible than phocid species. [ABSTRACT FROM AUTHOR]

Copyright of Agroindustria, Sociedad y Ambiente is the property of Revista Cientifica Agroindustria, Sociedad y Ambiente, ASA and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Somatic embryogenesis (SE) has many applications in grapevine biotechnology including micropropagation, eradicating viral infections from infected cultivars, mass production of hypocotyl explants for micrografting, as a continuous source for haploid and doubled haploid plants, and for germplasm conservation. It is so far the only pathway for the genetic modification of grapevines through transformation. The single-cell origin of somatic embryos makes them an ideal explant for mutation breeding as the resulting mutants will be chimera-free. In the present research, two combinations of plant growth regulators and different explants from flower buds at two stages of maturity were tested in regard to the efficiency of callusing and embryo formation from the callus produced in three white grape cultivars. Also, the treatment of somatic embryos with the chemical mutagen ethyl methanesulfonate (EMS) was optimised. Medium 2339 supplemented with β-naphthoxyacetic acid (5 μM) and 6-benzylaminopurine (BAP—9.0 μM) produced significantly more calluses than medium 2337 supplemented with 2,4-dichlorophenoxyacetic acid (4.5 µM) and BAP (8.9 µM) in all explants. The calluses produced on medium 2337 were harder and more granular and produced more SEs. Although the stage of the maturity of floral bud did not have a significant effect on the callusing of the explants, calluses produced from immature floral bud explants in the premeiotic stage produced significantly more SEs than those from more mature floral buds. Overall, immature ovaries and cut floral buds exposing the cut ends of filaments, style, etc., tested for the first time in grapevine SE, produced the highest percentage of embryogenic calluses. It is much more efficient to cut the floral bud and culture than previously reported explants such as anthers, ovaries, stigmas and styles during the short flowering period when the immature flower buds are available. When the somatic embryos of the three cultivars were incubated for one hour with 0.1% EMS, their germination was reduced by 50%; an ideal treatment considered to obtain a high frequency of mutations for screening. Our research findings will facilitate more efficient SE induction in grapevines and inducing mutations for improving individual traits without altering the genetic background of the cultivar. [ABSTRACT FROM AUTHOR]

The Gymnocalycium genus includes numerous highly valued species in the ornamental plant market and their propagation is usually carried out using traditional methods. However, there is a lack of information regarding the efficiency of micropropagation through in vitro tissue culture techniques on these species. So, with the objective of establishing an efficient micropropagation protocol that allows for optimizing the plant obtaining processes, the morphogenic potential of Gymnocalycium cv. Fancy has been studied in this work. For this purpose, plants of two different sizes (medium and large) were used as the starting material, from which three types of explants were obtained (apex, central discs and bases). The effect of three plant growth regulators (6-Benzylaminopurine, BAP; Kinetin, KIN; and Thidiazuron, TDZ) at three different concentrations each were studied, and the number of generated shoots, the frequency of appearance of callogenesis and rhizogenesis by explant and condition were assessed. An efficient protocol based on the use of KIN at 4 µM and central discs as a starting explant was developed. Moreover, the obtained sprouts rooted successfully (especially using BAP at 2 µM), and their subsequent acclimatization was very effective. Furthermore, emergence of a new morphotype is presented, that has not previously been described. [ABSTRACT FROM AUTHOR]

Objective: to determine a disinfestation procedure for in vitro establishing of Agave angustifolia Haw. explants collected in the field. Design/Methodology/Approach: seven agave plants per treatment were placed in perlite to contrast six treatments; two agrochemicals (Benomyl 2 g L-1 and Actara 0.5 g L-1) and three contact times (10, 20 and 30 days). The stems of the plants were sectioned in 4-6 segments, after treatment with alcohol 80%, chlorine 30% and hydrogen peroxide 20%, to later place them in vitro. Usually there is contamination of explants mainly by bacteria, so Pursue® was used at 25 and 50%, placing 20 explants in each concentration for 1 minute. The variables evaluated were percentage of visibly healthy explants, type of contamination. Due to the difference in the number of explants, no statistical tests were performed. Results: the plants treated with Benomyl presented less contamination by fungi without considering the contact time. Bacteria were present in the different treatments used, however, in the explants with pretreated with Benomyl, 85% was obtained visibly healthy when Pursue® at 50% was used. Study limitations/Implications: other agrochemicals, such as specific bactericides are desirable to determine through their use, whether they mitigate in vitro contamination by pathogenic bacteria. Findings/Conclusions: pretreatments with agrochemicals for the in vitro establishing of plants collected from the field are necessary to increase the number of visually healthy explants. With the Pursue® product at 50%, up to 85% of explants can be obtained without the presence of fungi or bacteria. [ABSTRACT FROM AUTHOR]

Purpose: The widespread adoption of endovascular aneurysm repair (EVAR) as preferred treatment modality for abdominal aortic aneurysm (AAA) has enlarged the number of patients needing open surgical conversion (OSC). The relationship between adherence to Instructions For Use (IFU) and EVAR long-term outcomes remains controversial. The aim of this study is to compare preoperative differences and postoperative outcomes between EVAR patients not adjusted to IFU and adjusted to IFU who underwent OSC. Methods: This multicenter retrospective study reviewed 33 explanted EVARs between January 2003 and December 2019 at 14 Vascular Units. Patients were included if OSC occurred >30 days after implantation and excluded if explantation was performed to treat an endograft infection, aortic dissection, or traumatic transections. Variables analyzed included baseline characteristics, adherence to IFU, implant and explant procedural details, secondary reinterventions, and postoperative outcomes. Results: Fifteen explanted patients (15/33, 45.5%) were identified not accomplished to IFU (out-IFU) at initial EVAR vs 18 explanted patients adjusted (in-IFU). During follow-up, a mean of 1.73±1.2 secondary reinterventions were performed, with more type I endoleaks treated in the subgroup out-IFU: 16.7% vs 6.3% in-IFU patients and more type III endoleaks (8.3% vs 0%). Patients out-IFU had shorter mean interval from implant to explant: 47.60±28.8 months vs 71.17±48. Type II endoleak was the most frequent indication for explantation. Low-flow endoleaks (types II, IV, V) account for 44% of indications for OSC in subgroup of patients in-IFU, compared with 13.3% in patients out-IFU and high-flow endoleaks (types I and III) were the main indication for patients out-IFU (33.3% vs 16.7% in-IFU). Total endograft explantation was performed in 57.5% of cases (19/33) and more suprarenal clamping was required in the subgroup out-IFU. Overall, 30-day mortality rate was 12.1% (4/33): 20% for patients out-IFU and 5.6% in-IFU. Conclusions: In our experience, type II endoleak is the most common indication for conversion and differences have been found between patients treated outside IFU with explantation taking place earlier during follow-up, mainly due to high-flow endoleaks and with higher mortality in comparison with patients adjusted to IFU. Ongoing research is required to delve into these differences. [ABSTRACT FROM AUTHOR]

Mast cells are activated through a variety of different receptors to release preformed granules and mediators synthesized de novo. However, the physiology and function of mast cells are not fully understood. Traditional studies of mast cell activation in humans have utilized cultures of tissue-derived mast cells including CD34+ progenitor cells or well-characterized commercially available cell lines. One limitation of these methods is that mast cells are no longer in a natural state. Therefore, their applicability to human skin disorders may be limited. Human skin explant models have been utilized to investigate the short-term effects of cell mediators, drugs, and irritants on skin while avoiding the ethical concerns surrounding in vivo stimulation studies with non-approved agents. Nonetheless, few studies have utilized intact human tissue to study mast cell degranulation. This “Methods” paper describes the development and application of an intact skin explant model to study human mast cell activation. In this manuscript, we share our protocol for setting up ex vivo human skin explants and describe the results of stimulation experiments and techniques to minimize trauma-induced histamine release. Skin explants were generated using de-identified, full-thickness, non-diseased skin specimens from plastic and reconstructive surgeries. Results were reproducible and demonstrated FcɛRI- and MRGPRX2-induced mediator release which was inhibited with the use of a BTK inhibitor and QWF, respectively. Thus, this explant model provides a quick and accessible method of assessing human skin mast cell activation and inhibition.

Plantation forestry using highly productive and fast-growing plants involves the use of high-quality improved planting material for genetic constancy, which is solved by microclonal reproduction. However, the high survival rate and stability of plants obtained in vitro are realised as a result of post-aseptic adaptation of regenerants. Therefore, the research aims to improve the survival techniques of paulownia regenerants of the Ukrainian varieties “Feniks” and “Enerdzhy” ex vitro. The study was conducted in December 2021 on the Ukrainian paulownia varieties “Feniks” and “Enerdzhy”, cultivar “Feniks”. The survival rate, development of regenerants, and their damage by facultative saprophytic pathogens differed on substrates of organic (Eco Plus, La Flora, Jiffy, coconut peat, cotton wool) and mineral (vermiculite, perlite, sand) origin. The biological products “Rise P” and “Prestop” proved to be effective for the control of pathogenic microorganisms on substrates of organic origin. The technological feasibility of using perlite substrate is substantiated. The pathophysiology of ex vitro regenerants of paulownia under Fusarium is described. The thickness of the films used for sheltering influenced the regeneration, engraftment, microbial damage, and temperature of the wet chambers. Growing regenerants with covering films of different thicknesses showed a high survival rate in the variants with stretch films of 10 and 23 μm thickness. In the case of using polyethene films with thicknesses of 60, 80 and 100 μm, a sharp increase in temperature was observed, which was 48, 53 and 650C, respectively. At these high temperatures, most of the regenerants died, after which they were colonised by facultative saprophytes, and the tissues were completely macerated within two days. The optimal ratio of juvenility reduction and the acquisition of adaptive traits is characteristic of the second, third and fourth generation ex vitro. The expediency of propagation by stem cuttings up to the fourth generation, which preserves the regenerative ability of shoot cuttings, was substantiated. The practical results of the research are a protocol for growing paulownia plants in vitro and ex vitro on different types of substrates for use by institutions engaged in paulownia propagation

[Display omitted] [ABSTRACT FROM AUTHOR]

Chrysanthemum morifolium ‘11-C-2’ is a variety of chrysanthemums with high ornamental and tea value, experiencing significant market demand. However, as cultivation areas expand, issues such as viral infection, germplasm degradation, low proliferation coefficient, and slow proliferation rate arise, necessitating the establishment of an efficient in vitro regeneration system. This study, based on the principles of orthogonal experimental design, explored the regeneration system of Chrysanthemum cultivar ‘11-C-2’ using sterile seedlings. The research focused on three key stages: adventitious bud differentiation, rooting culture, and acclimatization–transplantation, employing shoot-bearing stem segments and leaves as explants. The findings indicate that the optimal explant for the Chrysanthemum ‘11-C-2’ sterile seedlings is the shoot-bearing stem segment. The best medium for adventitious bud differentiation was determined to be MS supplemented with 1.5 mg/L 6-BA and 0.5 mg/L NAA. Bud differentiation began on day 17 with a 100% differentiation rate, completing around day 48. The maximum differentiation coefficient reached 87, with an average of 26.67. The adventitious buds were then cultured for rooting in the optimal medium of 1/2 MS supplemented with 0.1 mg/L NAA. Rooting was initiated on day 4 and was completed by day 14, achieving a rooting rate of 97.62%. After a 5-day acclimatization under natural light, the rooted seedlings were transplanted into a growth substrate with a peat-to-vermiculite ratio of 1:2. The plants exhibited optimal growth, with a transplantation survival rate of 100%. The findings provide data support for the efficient large-scale propagation of ‘11-C-2’ and lay the foundation for germplasm preservation and genetic transformation research of tea chrysanthemums.

Abstract Background Basic leucine zipper (bZIP) proteins are important transcription factors in plants. To study the role of bZIP transcription factors in walnut explant browning, this study used bioinformatics software to analyze walnut bZIP gene family members, along with their transcript levels in different walnut tissues, to evaluate the transcriptional expression of this gene family during the primary culture of walnut explants and to reveal the mechanism of action of walnut bZIP genes in walnut explant browning. Results The results identified 65 JrbZIP genes in the walnut genome, which were divided into 8 subfamilies and distributed on 16 chromosomes. The results of transcriptome data analysis showed that there were significant differences in the expression of four genes, namely, JrbZIP55, JrbZIP70, JrbZIP72, and JrbZIP88, under both vermiculite and agar culture conditions. There were multiple hormone (salicylic acid, abscisic acid, auxin, and gibberellin) signaling and regulatory elements that are responsive to stress (low temperature, stress, and defense) located in the promoter regions of JrbZIP55, JrbZIP70, JrbZIP72, and JrbZIP88. The walnut JrbZIP55 protein and Arabidopsis bZIP42 protein are highly homologous, and the proteins interacting with Arabidopsis bZIP42 include the AT2G19940 oxidoreductases, which act on aldehyde or oxygen-containing donors. Conclusion It is speculated that JrbZIP55 may participate in the regulation of browning in walnut explants.

Abstract Background Mammary gland (MG) infections (mastitis) are frequent diseases of dairy cows that affect milk quality, animal welfare and farming profitability. These infections are commonly associated with the bacteria Escherichia coli and Staphylococcus aureus. Different in vitro models have been used to investigate the early response of the MG to bacteria, but the role of the teat in mastitis pathogenesis has received less attention. In this study, we used punch-excised teat tissue as an ex vivo model to study the immune mechanisms that arise early during infection when bacteria have entered the MG. Results Cytotoxicity and microscopic analyses showed that bovine teat sinus explants have their morphology and viability preserved after 24 h of culture and respond to ex vivo stimulation with TLR-agonists and bacteria. LPS and E. coli trigger stronger inflammatory response in teat when compared to LTA and S. aureus, leading to a higher production of IL-6 and IL-8, as well as to an up-regulation of proinflammatory genes. We also demonstrated that our ex vivo model can be applied to frozen-stored explants. Conclusions In compliance with the 3Rs principle (replacement, reduction and refinement) in animal experimentation, ex vivo explant analyses proved to be a simple and affordable approach to study MG immune response to infection. This model, which better reproduces organ complexity than epithelial cell cultures or tissue slices, lends itself particularly well to studying the early phases of the MG immune response to infection.

Background.Callus culture in vitro is a convenient model for studying physiological and biochemical manifestations at the cellular level, as well as for cell selection to stress factors. To obtain a callus culture, it is necessary to select the optimal ratio of exogenous hormonal growth regulators (GRs) in the nutrient medium. The purpose of the study is to determine the optimal ratios of GRs at the stage of introducing cucumber and radish into in vitro culture. Materials and methods. Sterile plants of cucumber (Cucumis sativus L., cv. Unity) and radish (Raphanus sativus L., cv. Red Giant) were taken as the object of study. The experiment included obtaining a callus culture of cucumber and radish from various explants (cotyledon leaves, hypocotyl and roots) on a nutrient medium with a mineral base according to Murashige-Skoog supplemented by mesoinositol (100 mg/l), glycine (1 mg/l), nicotinic acid (0.5 mg/l), as well as growth regulators 2.4-dichlorophenoxyacetic acid (2.4- D) and 6-benzylaminopurine (6-BAP) in various ratios. Results. A method for introducing cucumber and radish into in vitro culture is described. The role of the hormonal composition of the nutrient medium in callusogenesis was shown, the influence of different concentrations of GRs, auxin 2.4-D and cytokinin 6-BAP, on callus formation was revealed. For various explants of cucumber and radish, the optimal concentrations of used GRs were determined. Conclusions. The optimal composition of GRs (auxin 2.4-D and cytokinin 6-BAP, mg/l) in Murashige-Skoog medium was selected for callusogenesis on various explants of cucumber (2:2) and radish (4:0.5). A higher intensity of callusogenesis in cucumber on hypocotyl explants and no dependence of callusogenesis on the type of explantin radish were revealed.

Failure by the developing conceptus to secrete sufficient interferon tau (IFNT), required for maternal recognition of pregnancy (MRP), at the appropriate time is related to early pregnancy loss in cattle. We aimed to test the hypothesis that there is a dose- and time-dependent relationship between IFNT and the endometrial expression of key interferon-stimulated genes (ISGs) involved in the signalling cascade leading to MRP in cattle. Candidate genes were identified first through a bioinformatic approach, where integrated transcriptomic data from two previous studies were analyzed to identify endometrial genes induced by IFNT. Next, expression of selected candidate genes was investigated in vitro in endometrial explants. Endometrial explants collected from cows (n = 8) in the late luteal phase of the estrous cycle were cultured in medium without (control) or with recombinant ovine IFNT (1, 10, 100 ng/mL) for 6 h. Simultaneously, endometrial explants were cultured in medium containing 100 ng/mL IFNT for different time periods (15 min, 30 min, 1 h, 3 h, 6 h). Gene expression was analyzed by RT-qPCR. We identified 54 endometrial genes responding to IFNT and to some degree to the conceptus, from which five ISGs (CMPK2 , BPNT1 , IFI35 , TNFSF10 and TRIM38) were further selected for the dose- and time-dependent experiments. Classical ISGs (ISG15 , OAS1 , MX1 and MX2) were up-regulated (P < 0.05) in endometrium by 1 ng/mL IFNT. However, other selected ISGs (CMPK2 , BPNT1 , IFI35 , TNFSF10 and TRIM38) were induced only by higher concentrations (10 and 100 ng/mL) of IFNT (P < 0.05). In terms of duration of exposure, IFNT at 100 ng/mL induced a significant (P < 0.05) increase in ISG15 and CMPK2 expression after 1 h incubation, while all other studied ISGs in the endometrium were upregulated when cultured for 3 or 6 h, but did not affect expression when the duration of culture was for 1 h or less. These results suggest that IFNT acts on the uterus in both a dose- and time-dependent manner in cattle and that timely exposure of the endometrium to sufficient IFNT is essential for appropriate signalling to ensure successful pregnancy establishment. • We identified 54 genes responding to IFNT in the endometrium around maternal recognition of pregnancy in cattle. • Classical ISGs (ISG15 , OAS1 , MX1 , MX2) were upregulated in the endometrium by 1 ng/mL IFNT. • Other selected ISGs (CMPK2 , BPNT1 , IFI35 , TNFSF10 , TRIM38) were only induced at higher concentrations of IFNT (10 and 100 ng/mL). • IFNT at 100 ng/mL induced all studied ISGs in the endometrium when cultured for 3 or 6 h but not for a shorter period. [ABSTRACT FROM AUTHOR]

Plantation forestry using highly productive and fast-growing plants involves the use of high-quality improved planting material for genetic constancy, which is solved by microclonal reproduction. However, the high survival rate and stability of plants obtained in vitro are realised as a result of post-aseptic adaptation of regenerants. Therefore, the research aims to improve the survival techniques of paulownia regenerants of the Ukrainian varieties "Feniks" and "Enerdzhy" ex vitro. The study was conducted in December 2021 on the Ukrainian paulownia varieties "Feniks" and "Enerdzhy", cultivar "Feniks". The survival rate, development of regenerants, and their damage by facultative saprophytic pathogens differed on substrates of organic (Eco Plus, La Flora, Jiffy, coconut peat, cotton wool) and mineral (vermiculite, perlite, sand) origin. The biological products "Rise P" and "Prestop" proved to be effective for the control of pathogenic microorganisms on substrates of organic origin. The technological feasibility of using perlite substrate is substantiated. The pathophysiology of ex vitro regenerants of paulownia under Fusarium is described. The thickness of the films used for sheltering influenced the regeneration, engraftment, microbial damage, and temperature of the wet chambers. Growing regenerants with covering films of different thicknesses showed a high survival rate in the variants with stretch films of 10 and 23 μm thickness. In the case of using polyethene films with thicknesses of 60, 80 and 100 μm, a sharp increase in temperature was observed, which was 48, 53 and 650C, respectively. At these high temperatures, most of the regenerants died, after which they were colonised by facultative saprophytes, and the tissues were completely macerated within two days. The optimal ratio of juvenility reduction and the acquisition of adaptive traits is characteristic of the second, third and fourth generation ex vitro. The expediency of propagation by stem cuttings up to the fourth generation, which preserves the regenerative ability of shoot cuttings, was substantiated. The practical results of the research are a protocol for growing paulownia plants in vitro and ex vitro on different types of substrates for use by institutions engaged in paulownia propagation. [ABSTRACT FROM AUTHOR]

The management of chronic heart failure over the past decade has witnessed tremendous strides in medical optimization and device therapy including the use of left ventricular assist devices (LVAD). What we once thought of as irreversible damage to the myocardium is now demonstrating signs of reverse remodeling and recovery. Myocardial recovery on the structural, molecular, and hemodynamic level is necessary for sufficient recovery to withstand explant and achieve sustained recovery post‐LVAD. Guideline‐directed medical therapy and unloading have been shown to aid in recovery with the potential to successfully explant the LVAD. This review will summarize medical optimization, assessment for recovery, explant methodologies and outcomes post‐recovery with explant of durable LVAD. [ABSTRACT FROM AUTHOR]