Search Results (180)

Genetic transformation is an essential tool for investigating gene function and editing genomes. Kiwifruit, recognized as a significant global fresh fruit crop, holds considerable economic and nutritional importance. However, current genetic transformation techniques for kiwifruit are impeded by low efficiency, lengthy culture durations (a minimum of six months), and substantial labor requirements. In this research, we established an efficient system for shoot regeneration and the stable genetic transformation of the ‘Hayward’ cultivar, utilizing leaf explants in conjunction with two strains of Agrobacterium that harbor the expression vector pBI121-35S::GFP, which contains the green fluorescent protein (GFP) gene as a visible marker within the T-DNA region. Our results show that 93.3% of leaf explants responded positively to the regeneration medium, producing multiple independent adventitious shoots around the explants within a six-week period. Furthermore, over 71% of kanamycin-resistant plantlets exhibited robust GFP expression, and the entire transformation process was completed within four months of culture. Southern blot analysis confirmed the stable integration of GFP into the genome, while RT-PCR and fluorescence microscopy validated the sustained expression of GFP in mature plants. This efficient protocol for regeneration and transformation provides a solid foundation for micropropagation and the enhancement of desirable traits in kiwifruit through overexpression and gene silencing techniques. Full article

Roses are one of the most essential ornamental flowers in the world. At present, traditional techniques such as cross breeding are mainly used in rose breeding. The inefficiency of the in vitro regeneration system has become the limiting step for the innovation and genetic improvement of rose germplasm resources. A tissue culture rapid propagation system of Rosa ‘Pompon Veranda’ was established using the stem segments with shoots as the initial experimental material. The results showed that the best disinfection method was to soak the explants in 75% ethanol for 1 min, and then soak them in 15% sodium hypochlorite solution for 15 min. The contamination rate was only about 6%. The best rooting medium for tissue culture seedlings was 1/2MS with 0.1 mg∙L⁻¹ NAA, and the rooting rate can reach around 95%. On this basis, calluses were induced by using leaflets of tissue-cultured seedlings as explants. The results showed that the optimal medium for inducing callus tissue was MS + 5.0 mg∙L⁻¹ 2,4-D, with an induction rate of 100%. The calluses were cultured in the medium of MS with 0.01 mg∙L⁻¹ NAA, 1.5 mg∙L⁻¹ TDZ and 0.1 mg∙L⁻¹ GA₃ for 12 days in the dark and then transferred to light conditions. The differentiation rate of callus was 10.87%. On the medium of MS with 0.5 mg∙L⁻¹ 6-BA, 0.004 mg∙L⁻¹ NAA and 0.1 mg∙L⁻¹ GA₃, the shoots could regenerate into whole plants. This study has established an in vitro regeneration system of R. ‘Pompon Veranda’, which is a key perquisite for the subsequent establishment of its genetic transformation system. Moreover, this method will also be an important reference for studies on quality traits such as floral scent and prickles of Rosa plants. Full article

Introduction: Two-stage revision with an antibiotic-loaded, temporary static cement spacer is a common treatment for periprosthetic joint infection (PJI) of the knee. However, limited data exists on in vivo antibiotic elution kinetics after spacer implantation. This pilot study uses the technique of microdialysis (MD) to collect intra-articular knee samples. The aim was to evaluate MD as an intra-articular sampling method to detect spacer-eluted antibiotics within 72 h after surgery and to determine whether they show specific elution kinetics. Methods: Ten patients (six male, four female; age median 71.5 years) undergoing two-stage revision for knee PJI were included. A MD catheter was inserted into the joint during explantation of the infected inlying implant and implantation of a custom-made static spacer coated with COPAL cement (0.5 g gentamicin (G) and 2 g vancomycin (V)). Over 72 h postoperatively, samples were collected and analyzed for spacer-eluted antibiotics, intravenously administered antibiotics (e.g., cefazolin and cefuroxime), metabolic markers (glucose and lactate), and Interleukin-6 (IL-6). Local and systemic levels were compared. Results: All catheters were positioned successfully and well tolerated for 72 h. Antibiotic concentrations in MD samples peaked within the first 24 h (G: median 9.55 µg/mL [95% CI: 0.4–17.36]; V: 37.57 µg/mL [95% CI: 3.26–81.6]) and decreased significantly over 72 h (for both p < 0.05, G: 4.27 µg/mL [95% CI: 2.26–7.2]; V: 9.69 µg/mL [95% CI: 3.86–24]). MD concentrations consistently exceeded blood levels (p < 0.05), while intravenously administered antibiotics showed higher blood concentrations. Glucose in MD samples decreased from 17.71 mg/dL to 0.89 mg/dL (p < 0.05). IL-6 and lactate concentrations showed no difference between MD and blood samples. Conclusions: Monitoring antibiotics eluted by a static spacer with intra-articular MD for 72 h is feasible. Gentamicin and vancomycin levels remained above the minimal inhibitory concentration. Differentiating infection from surgical response using metabolic and immunological markers remains challenging. Prolonged in vivo studies with MD are required to evaluate extended antibiotic release in two-stage exchanges. Full article

Background: Cochlear implantation is widely used to provide auditory rehabilitation to individuals with severe-to-profound sensorineural hearing loss. However, electrode insertion during cochlear implantation leads to inner ear trauma, damage to sensory structures, and consequently, loss of residual hearing. There is very limited information regarding the target proteins involved in electrode insertion trauma (EIT) following cochlear implantation. Methods: The aim of our study was to identify target proteins and host molecular pathways involved in cochlear damage following EIT utilizing the iTRAQ™ (isobaric tags for relative and absolute quantification) technique using our ex vivo model. The organ of Corti (OC) explants were dissected from postnatal day 3 rats and subjected to EIT or left untreated (control). The proteins were extracted, labelled, and subjected to ultra-high performance liquid chromatography–tandem mass spectrometry. Results: We identified distinct molecular pathways involved in EIT-induced cochlear damage. Confocal microscopy confirmed the expression of these identified proteins in OC explants subjected to EIT. By separating the apical, middle, and basal cochlear turns, we deciphered a topographic array of host molecular pathways that extend from the base to the apex of the cochlea, which are activated post-trauma following cochlear implantation. Conclusions: The identification of target proteins involved in cochlear damage will provide novel therapeutic targets for the development of effective treatment modalities for the preservation of residual hearing in implanted individuals. Full article

Orchids and other flowering plants offer a wide range of floral traits. Within the Orchidaceae family, the Vanilla genus is one of the most valued plants in the commercial flavor industry. In vitro biotechnological approaches to Vanilla, such as germplasm conservation, massive propagation, and genetic engineering, have played a key role in breeding programs. There are, however, few studies that elucidate the physiological, molecular, and genetic aspects of vanilla orchid flowering and in vitro induction. This review’s main objective is to provide updated and complete data on in vitro floral induction and flowering of tropical and vanilla orchid species. A bibliographic search was carried out for scientific reports in academic databases (Scopus, Web of Science, PubMed, and ScienceDirect), and a total of 39 documents from 2014 and 2025 were analyzed. This review discusses the most important factors that affect in vitro flowering in Vanilla, including the monopodial genotypes, photoperiod, irradiance, temperature, nutrition, plant growth regulators, explant types, and culture methods. Consequently, this revision incorporates a number of studies on orchid in vitro flowering, with a focus on vanilla species. In conclusion, there still exists limited progress in Vanilla compared to other orchid species; however, the use of biotechnological techniques like in vitro flowering offers a fundamental framework for orchid breeding. Full article

Background: Infective endocarditis after transcatheter aortic valve replacement (TAVR-IE) is a rare but severe complication associated with high morbidity and mortality. The optimal treatment strategy—surgical explantation versus medical therapy—remains uncertain, particularly given the technical demands of TAVR removal and the advanced age of many affected patients. Methods: We conducted a systematic review and meta-analysis of studies comparing the surgical and medical management of TAVR-IE. Primary outcomes included 30-day mortality and 1-year survival. Secondary analyses explored microbiological profiles, patient demographics, prosthesis type, postoperative complications, and surgical indications. A qualitative synthesis of surgical explantation techniques and reconstructive strategies was also performed based on recent consensus recommendations. Results: Three studies comprising 1557 patients with TAVR-IE were included; 155 (10.0%) underwent surgical treatment. Thirty-day mortality was comparable between groups (surgical: 9.7%; medical: 8.4%), while the pooled odds ratio for one-year survival did not reach statistical significance (OR: 1.91, 95% CI: 0.36–10.22; I² = 88%). However, single-center outcomes demonstrated markedly improved survival with surgery (96% vs. 51%). The most common surgical indications included severe valvular dysfunction (50.3%), aortic root abscess (26.5%), and large vegetations (21.3%), in line with current guideline recommendations. Postoperative complications included acute renal failure (10%) and longer hospitalizations (19.8 vs. 18 days), although these were not statistically different. Contemporary explant strategies—such as the Double Kocher, Tourniquet, and Y-incision aortic enlargement techniques—were highlighted as critical tools for surgical success. Conclusions: While underutilized, surgical intervention for TAVR-IE may offer significant survival benefits in select patients, particularly when guided by established indications and performed at high-volume centers. Outcomes depend heavily on timing, surgical expertise, and appropriate patient selection. As TAVR expands to younger populations, TAVR-IE will become increasingly relevant, necessitating early multidisciplinary involvement and broader familiarity with advanced explant techniques among cardiac surgeons. Full article

Passiflora caerulea L., commonly known as the blue passionflower, is traditionally grown as an ornamental plant, but has a diverse chemical composition resulting in a wide range of biological activities that determine its pharmacological properties and use in medicine. Traditional propagation methods, including seed germination and vegetative cuttings, are often inefficient due to low germination rates, susceptibility to pathogens, and slow growth. In particular, P. caerulea presents significant challenges in germination due to its slow development. In this context, in vitro cultivation is used to enable rapid, large-scale plant production while maintaining genetic fidelity. The study of Passiflora tissue cultures began in 1966 and has since attracted increasing attention from researchers around the world. However, despite growing interest, studies specifically focused on the in vitro propagation of P. caerulea remain limited. This review aims to summarize existing knowledge on the main techniques used for in vitro culturing and propagation of P. caerulea, including organogenesis, somatic embryogenesis, and callogenesis. Particular attention is paid to the key factors that influence the initiation, growth, and regeneration of cultures, including the type of explant, the composition of the media, and the environmental conditions. Advances in the in vitro cultivation of P. caerulea have greatly improved the understanding and propagation of this species. Although in vitro cultivation offers several advantages, it is crucial to conduct thorough research on the selection of explants, their age, and the appropriate culture media to ensure optimal growth and development. Full article

Black elder (Sambucus nigra L.) is a temperate shrub with flowers and fruits that are edible after processing. This species is not yet widely known in the global agricultural sector, but its adaptability and drought tolerance may generate more interest in this crop. Our study aimed to find suitable micropropagation techniques for the black elder ‘Albida’ and compare suitable statistical methods for evaluating multiplication and rooting. For micropropagation, we tested the Murashige and Skoog (MS) growth medium with selected auxins and cytokinins. Five proliferation MS media containing 1, 2, and 4 mg/L BAP or 0.5 and 1 mg/L TDZ were tested. To induce root formation, three types of auxins were tested at a concentration of 1 mg/L in a 50% MS medium: IBA, IAA, and NAA. Data analysis was performed using different parametric and nonparametric tests to robustly capture the effects of treatments across varying distributional scenarios in developing explants subjected to the interactions of internal native and externally added plant growth regulators. The average multiplication rate ranged from 1.6 to 2.0 shoots per explant. High multiplication was recorded on the MS medium with 1 mg/L 6-benzylaminopurine. The root number per rooted explant was highly variable, ranging from 3.0 to 12.0 roots per explant. The highest average root number result was observed when 1 mg/L α-naphthalenacetic acid was used. All rooted plants were successfully acclimated to normal growing conditions. This in vitro propagation protocol allows for the production of hundreds to thousands of rooted plants from one initial explant within one year, enabling faster introduction to the agronomic sector. Full article

The introduction of new ornamental species and cultivars is one of the hallmarks of innovation in global floriculture. Brunfelsia uniflora, a subshrub native to Brazil, has white, lilac, and blue flowers on the same plant, in addition to a distinctive fragrance. As it is a wild species, technologies such as large-scale clonal propagation of superior genotypes are still scarce, limiting its supply to the flower market. Therefore, a successful micropropagation protocol was developed for B. uniflora using nodal segments and shoot tips as initial explants. In the multiplication phase, the use of 6-benzylaminopurine produced the highest multiplication rates (10.3–10.9 shoots/explant) and the number of leaves in the shoots. In vitro shoot rooting using MS medium with reduced macronutrient concentrations and supplemented with IBA resulted in a 91.7% rooting rate. The greatest difficulty in micropropagating this species was the high percentage of shoots that developed calli. The highest percentage of callus formation occurred with the addition of auxins at high concentrations (1.0 and 1.5 mg L⁻¹). Even so, the shoots and plantlets were acclimatized, demonstrating the effectiveness of this technique for the production of B. uniflora plantlets. Full article

Philodendron ‘White Knight’ is a popular climbing evergreen plant typically propagated through stem cuttings. However, this method is slow and inefficient, making it challenging to meet the rising market demand. In vitro propagation could enhance the multiplication of this cultivar. However, research on its in vitro propagation is limited. Therefore, the objective of the present study was to establish an efficient micropropagation technique to mass-produce Philodendron ‘White Knight’ to meet the market demand. We investigate the impact of silver nanoparticles (Ag NPs) on the surface sterilization of Philodendron ‘White Knight’ petioles, the role of plant growth regulators in adventitious shoot regeneration and shoot multiplication, and the effect of auxins on the rooting ability of Philodendron ‘White Knight’ microshoots. There are few stages in plant micropropagation. The establishment of aseptic culture is the first and most important stage. For Philodendron ‘White Knight’, aseptic petiole explants (100%) were obtained after treatment with 40 mg L⁻¹ Ag NPs for 60 min. This was followed by adventitious shoot induction, and the highest rate of adventitious shoot induction (52.6%) and the maximum shoot number (13.9 shoots per petiole) were achieved on Murashige and Skoog shoot multiplication B (MS-B) medium with 20 µM of 2-isopentenyl adenine (2-IP) and 5.0 µM of naphthalene acetic acid (NAA). The shoot multiplication stage was achieved with the highest number of shoots (34 shoots per shoot tip) with a length of 5.1 cm, which was obtained on MS-B medium with 5.0 µM 2-IP and 2.5 µM NAA. All the microshoots produced roots during the root induction stage with the maximum root number (8.2 roots per shoot), and the greatest plantlet height (9.1 cm) was achieved on half-strength Murashige and Skoog medium containing indole-3-butyric acid (10.0 μM). The rooted plantlets of Philodendron ‘White Knight’ were transplanted into a substrate composed of 10% peat moss, 50% orchid stone, and 40% coconut husk chips and acclimatized in a greenhouse environment, achieving a survival rate of 100%. This micropropagation protocol can be used for the commercial production of Philodendron ‘White Knight’. Full article

Hyaluronic acid (HA) is an unbranched polysaccharide particularly abundant in the extracellular matrix (ECM) of soft connective tissues. In humans, about 50% of the total HA in the organism is localized in the skin. HA plays an essential role in the hydration of the ECM, in the regulation of tissue homeostasis, in the resistance to mechanical stimuli/forces, and in the modulation of tissue regeneration. For these reasons, HA is widely used in regenerative medicine and cosmetics. In this study we used an innovative fluid dynamic system to investigate the effects of a cross-linked macrostructural HA formulation on dermal collagen of healthy human skin explants. The good preservation of skin explants provided by the bioreactor allowed applying refined high-resolution microscopy techniques to analyze in situ the HA-induced modifications on the ECM collagen fibrils up to 48 h from the application on the skin surface. Results demonstrated that this HA formulation, commercially proposed for subcutaneous injection, may act on dermal ECM also when applied transcutaneously, improving ECM hydration and modifying the organization of the collagen fibrils. These findings, obtained by the original combination of explanted human skin use with an advanced culture system and multiscale imaging techniques, are consistent with the volumizing and anti-aging effect of HA. Full article

Vaccinium floribundum Kunth, also known as mortiño, is of cultural, gastronomic, pharmaceutical and ecological importance in the Andes due to its regenerative capacity to preserve vegetation after destructive fires. The main limitation for the production of mortiño fruits is that the plant has not been domesticated or cultivated, which could pose risks to the species and the paramos where it lives. In vitro culture is a crucial technique for propagating horticultural crops where factors such as the concentration, growth regulators, medium and explant parameters must be optimized to ensure the success of in vitro propagation techniques. This review uses the Prisma methodology, identifying 47 studies on the in vitro cultivation of Vaccinium, but only five studies on the domestication of V. floribundum Kunth using three in vitro cultivation techniques (axillary buds, seed germination and induced callogenesis) were published in Scopus and ScienceDirect. Therefore, the objective is to provide information on in vitro propagation techniques for the domestication of V. floribundum Kunth. Full article

Caragana intermedia is a perennial shrub species in the genus Caragana (Fabaceae), demonstrating remarkable stress resistance and adaptability. However, research on its somatic embryogenesis (SE) and genetic transformation techniques remains limited. In this study, we established an SE system by utilizing immature cotyledons isolated from young C. intermedia seeds. Our findings demonstrated that the immature cotyledons at 6–7 weeks after flowering (WAF) were the best explants for SE. The optimal embryo induction medium consisted of an MS basal medium supplemented with 5 mg/L α-naphthaleneacetic acid (NAA), 3 mg/L 6-benzylaminopurine (6-BA), 30 g/L sucrose, 7 g/L agar, and 500 mg/L hydrolyzed casein. Cotyledon-stage embryos germinated on a half-strength MS medium, exhibiting a 34.36% germination rate. Based on the SE system, we developed a preliminary genetic transformation system using the RUBY reporter gene, which successfully generated transgenic calli and cotyledon-stage embryos. The establishment of the SE system is expected to shorten breeding cycles, facilitate propagation of superior cultivars, and support large-scale industrial applications in C. intermedia. Furthermore, the stable transformation system provides a platform for molecular breeding and gene function verification. Full article

In this study, a reliable and efficient micropropagation protocol was developed for Stachys byzantina, a valuable and promising ornamental species. For the initial in vitro cultures on the Murashige and Skoog (MS) medium, shoot tips were used as explants. The addition of 5 μM of kinetin (KIN) resulted in the production of multiple (6.0 shoots/explant) and elongated (3.6 cm) shoots. The MS medium supplemented with 10 μM of a-Naphthaleneacetic acid (NAA) proved efficient for the in vitro rooting (73.3%) of the microshoots. For the ex vitro rooting of the microshoots, the treatment with 0.5 g L⁻¹ of Indole-3-butyric acid potassium salt (K-IBA), before planting in 1:1 (v/v) peat and perlite substrate and placed in a fog system, led to 86.7% rooting. The acclimatization stage was successful, and 96.7% survival was recorded for the ex vitro-rooted plantlets. Inter Simple Sequence Repeat (ISSR) markers were employed to examine the genetic uniformity of the in vitro-derived plantlets with the mother S. byzantina plants. The monomorphic banding pattern in the micropropagated plants and the mother plant confirmed the genetic uniformity of the in vitro-derived plantlets and revealed the reliability of the proposed in vitro protocol for S. byzantina. As far as we know, this is the first study on a combined micropropagation and genetic uniformity assessment of the species, the findings of which could be further used to apply new in vitro cultivation techniques or to produce elite genotypes of S. byzantina. Full article

Background/Objectives: Transcatheter Aortic Valve Implantation (TAVI) has significantly improved the management of aortic valve disease, but post-TAVI infective endocarditis, occurring in 0.5–3.1% of cases, remains a serious complication. Due to a high mortality rate and technical challenges, surgical replacement of infected TAVI prosthetic valves is performed in only 11.4% of cases. Methods: This case describes a standardized surgical technique for the removal and replacement of self-expanding TAVI prosthetic valves in the case of infective endocarditis. Results: The proposed approach aims to facilitate valve explantation while minimizing surgical risks. Conclusions: We believe that this technique could be particularly beneficial for surgeons managing these complex cases, by reducing surgical complications and improving patient outcomes. Further studies are necessary to validate its long-term efficacy and applicability in broader clinical settings. Full article