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Glioblastoma (GBM) is a highly aggressive adult brain cancer, characterised by poor prognosis and a dismal five-year survival rate. Despite significant knowledge gains in tumour biology, meaningful advances in patient survival remain elusive. The field of neuro-oncology faces many disease obstacles, one being the paucity of faithful models to advance preclinical research and guide personalised medicine approaches. Recent technological developments have permitted the maintenance, expansion and cryopreservation of GBM explant organoid (GBO) tissue. GBOs represent a translational leap forward and are currently the state-of-the-art in 3D in vitro culture system, retaining brain cancer heterogeneity, and transiently maintaining the immune infiltrate and tumour microenvironment (TME). Here, we provide a review of existing brain cancer organoid technologies, in vivo xenograft approaches, evaluate in-detail the key advantages and limitations of this rapidly emerging technology, and consider solutions to overcome these difficulties. GBOs currently hold significant promise, with the potential to emerge as the key translational tool to synergise and enhance next-generation omics efforts and guide personalised medicine approaches for brain cancer patients into the future. [ABSTRACT FROM AUTHOR]

Medicinal plants are integral to traditional medicine systems worldwide, being pivotal for human health. Harvesting plant material from natural environments, however, has led to species scarcity, prompting action to develop cultivation solutions that also aid conservation efforts. Biotechnological tools, specifically plant tissue culture and genetic transformation, offer solutions for sustainable, large-scale production and enhanced yield of valuable biomolecules. While these techniques are instrumental to the development of the medicinal plant industry, the challenge of inherent regeneration recalcitrance in some species to in vitro cultivation hampers these efforts. This review examines the strategies for overcoming recalcitrance in medicinal plants using a holistic approach, emphasizing the meticulous choice of explants (e.g. embryonic/meristematic tissues), plant growth regulators (e.g. synthetic cytokinins), and use of novel regeneration-enabling methods to deliver morphogenic genes (e.g. GRF/GIF chimeras and nanoparticles), which have been shown to contribute to overcoming recalcitrance barriers in agriculture crops. Furthermore, it highlights the benefit of cost-effective genomic technologies that enable precise genome editing and the value of integrating data-driven models to address genotype-specific challenges in medicinal plant research. These advances mark a progressive step towards a future where medicinal plant cultivation is not only more efficient and predictable but also inherently sustainable, ensuring the continued availability and exploitation of these important plants for current and future generations. [ABSTRACT FROM AUTHOR]

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

Abstract Objective The African bush apple (Heinsia crinita) is a highly valued orphan shrub that supports the nutritional and natural medicine needs of many sub-Saharan African communities. However, the crop remains poorly conserved and without any known genetic improvement. Accordingly, the current study sought to develop for the first time, an in vitro regeneration system based on direct shoot proliferation from its stem and hypocotyledonary explants using combinations of two cytokinins (benzyl adenine – BA, thidiazuron – TDZ) and the auxin (naphthalene acetic acid (NAA), in Murashige and Skoog (MS) medium. Results Combinations of BA and NAA effectively induced multiple shoot formation from stem and hypocotyledonary explants of the crop. The most effective treatment (1.0 mg/L BA + 0.1 mg/L NAA) induced an average of 13.77 and 30.32 shoots per responsive hypocotyl and stem explants, respectively. Combinations of TDZ and NAA were less effective in promoting shoot induction in the explants at the concentrations tested compared to BA and NAA combinations. Hypocotyledonary explants achieved complete plant regeneration without multiple shoot formation in a hormone-free MS medium. In vitro shoots regenerated from both stem and hypocotyledonary explants were 100% successfully rooted on a half-strength hormone-free medium, and acclimatized to produce H. crinita plants with over 90.0% efficiency.

Plant growth regulators (PGRs) control signaling networks and developmental processes involved in plant responses to various biotic and abiotic stresses, making it crucial to study PGRs in vitro . The protocol for micropropagation of Solanum lycopersicum L., following callus induction and regeneration through explants such as internodal segments, leaves, and nodal segments, was established during the present study. Explants were inoculated on Murashige and Skoog (MS) medium supplemented with different plant growth regulators like BA (6-benzylaminopurine), 2,4-D (2,4-dichlorophenoxyacetic acid), BA + 2,4-D, IAA (Indoleacetic acid), IBA (Indolebutyric acid), and NAA (Naphthaleneacetic acid). It was observed that among all explants, the nodal segment showed maximum callus induction (83.33%) and multiplication (86.67%) at 0.25 mg/l of 2,4-D; the highest shoot number (3.33) at 0.5 mg/l of IAA + 0.5 mg/l of BA; the greatest shoot length (7.57 cm) at 0.75 mg/l of BA; root induction (80.95%), root number (21.88), and root length (10.62 cm) at 1 mg/l of IAA. Additionally, the maximum fresh weight (2.448 g) was observed at 0.5 mg/l of BA, while the highest dry weight (0.172 g) and dry matter content (14.25%) were noted at 1 mg/l of BA + 1 mg/l of 2,4-D for the internodal segment. Results of the present study revealed that among different explants, the best response was given by nodal segments, followed by internodal segments. Among the different PGRs, 2,4-D resulted in the highest callus induction and multiplication percentage.

Objective: The African bush apple (Heinsia crinita) is a highly valued orphan shrub that supports the nutritional and natural medicine needs of many sub-Saharan African communities. However, the crop remains poorly conserved and without any known genetic improvement. Accordingly, the current study sought to develop for the first time, an in vitro regeneration system based on direct shoot proliferation from its stem and hypocotyledonary explants using combinations of two cytokinins (benzyl adenine – BA, thidiazuron – TDZ) and the auxin (naphthalene acetic acid (NAA), in Murashige and Skoog (MS) medium. Results: Combinations of BA and NAA effectively induced multiple shoot formation from stem and hypocotyledonary explants of the crop. The most effective treatment (1.0 mg/L BA + 0.1 mg/L NAA) induced an average of 13.77 and 30.32 shoots per responsive hypocotyl and stem explants, respectively. Combinations of TDZ and NAA were less effective in promoting shoot induction in the explants at the concentrations tested compared to BA and NAA combinations. Hypocotyledonary explants achieved complete plant regeneration without multiple shoot formation in a hormone-free MS medium. In vitro shoots regenerated from both stem and hypocotyledonary explants were 100% successfully rooted on a half-strength hormone-free medium, and acclimatized to produce H. crinita plants with over 90.0% efficiency. [ABSTRACT FROM AUTHOR]

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

• Nodal explants can be used to micropropagate P. longum for ex-situ conservation. • Thidiazuron was the most efficient hormone for regenerating shoots. • Indole-3-butyric acid was the most efficient hormone for regenerating roots. • First report on genetic fidelity of mother and in vitro -raised P. longum plants. • No polymorphisms found between mother and in vitro plants for all tested markers. Piper longum L. has been widely used to treat digestive disorders, insomnia, diabetes, bronchitis, and cough since ancient times, which has caused a decline in its population in natural habitat. This study aims to develop a protocol for micropropagation in P. longum using direct organogenesis using nodal explant as well as to compare genetic uniformity between the mother plant and in vitro -grown plants using Random Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeats (ISSR), and Start Codon Targeted (SCoT) markers. Nodal explants were cultured on Murashige & Skoog (MS) medium added with α-naphthaleneacetic acid (NAA), Kinetin (KN), Thidiazuron (TDZ), 6-Benzylaminopurine (BAP), and 10 % coconut water. The highest number of shoots (5.33±1.15) and longest shoot length (6.16±0.65 cm) were found in MS media added with 1.0 mg/L TDZ, while the highest number of roots (7.0 ± 1.0) and the longest root length (5.53±0.35 cm) were found in MS media added with 1.0 mg/L Indole-3-butyric acid (IBA) in nodal explants. The evaluation of genetic homogeneity using 12 specific RAPD, ISSR, and SCoT markers revealed that the in vitro -raised plants are genetically very similar, or even identical, to the wild mother plants, and there is no polymorphism between the mother plant and in vitro -raised plants for the specific markers tested. Twelve of twenty-four RAPD, ISSR, and SCoT primers amplified a total of 39 loci varying in size from 150 to 1400 base pairs. This study showed that an effective protocol developed for the in vitro propagation of P. longum by direct organogenesis can be used to produce true-to-type plants, which may aid in homozygous breeding programs and ex-situ conservation and could be used for commercial propagation. [ABSTRACT FROM AUTHOR]

A variation of the clonal propagation system for tea has been established through in vitro methods, utilizing shoots and leaves derived from primary explants of mature field-grown plants. The research aimed to assess the diversity of tea calluses based on morphological and phytochemical traits derived from leaf and shoot explants on MS media induced by the application of plant growth regulators, including auxin and cytokinin. The research was conducted at the Seed Technology Unit Tissue Culture Laboratory, Faculty of Agriculture, Jatinangor, from April 2022 to February 2023. The experiment employed MS-modified multiplication media and PGR, with BAP at 9 mg/L, TDZ at 1.0 mg/L, Zeatin at 0.1 mg/L, NAA at 0.01 mg/L, and 1 mg/L. Results indicated that applying PGR, auxin, and cytokinin with MS media influenced morphological and phytochemical traits of tea's callus derived from leaves and shoots. Specifically, the treatment of 1 mg/L NAA + 1 mg/L BAP showed the best performance on callus weight and total phenolic and flavonoid contents compared to other treatments., i.e. 9.038% % and 0.05% %, respectively. The success of clonal propagation in tea through in vitro methods is expected to increase by carefully selecting the appropriate source and type of explant, planting media, PGR, and addressing potential contaminants. [ABSTRACT FROM AUTHOR]

Silybum marianum L. is considered one of the most extensively used medicinal plants worldwide due to its therapeutic benefits. While ultrasound waves were used to enhance the properties of numerous plant species. However, no study was investigated on applying ultrasonic waves to this particular plant. Therefore, the study amid to assess the impact of different exposure periods (0,10, 20, 25, 30, 35, and 40 minutes) of a frequency of 47.6 KHz on callus induction, protein content, and plant regeneration in S. marianum. The effect of ultrasonication was distinctive in accelerating callus induction of S. marianum, especially in short exposing periods (10 and 20 minutes). The percentage of callus formation reached 100%, 83.3% for cotyledons and stem respectively, when exposed to 20 minutes of ultrasound. Additionally, growth and total protein content were increased at 40 days and 80 days post- treatments. In contrast, long exposing periods (30, 35, and 40 minutes) had a negative impact on callus induction from all explants, as well as on callus growth and protein content. Moreover, ultrasonication stimulated one-step shoot regeneration during callus induction. The percentage of this phenomenon reached 100% for cotyledon at 10 and 20 minutes exposing period. This study confirmed the advantages of applying ultrasonic waves, particularly during shorter period to enhance the cultivation of S. marianum in vitro.

'Jayanti Agrihorti' is a superior chrysanthemum therefore rooted cuttings quality is required. In vitro propagation increases the rate of propagation and produces true-to-type plants. The research to obtain the best explants and culture medium composition that is capable of producing high-quality plants. The research was arranged in a randomized complete block design consisting of two factors. The first factor was the explant, including the apical shoot of 0.5 cm, 1.0 cm, and the nodal segment. The second factor was in vitro culture medium composition (CP), consisting of CP1 (Murashige and Skoog (MS) medium for initiation, followed by MS + 2.5 mg.L⁻¹ gibberellic acid (GA₃) for subculture), CP2 (MS + 0.25 mg.L⁻¹ benzyl amino purine (BAP) for initiation, followed by MS for subculture), CP3 (MS + 0.25 mg.L⁻¹ BAP for initiation, followed by MS + 2.5 mg.L⁻¹ GA₃ for subculture), and CP4 (MS + 0.5 mg.L⁻¹ BAP for initiation, followed by MS + 0.25 mg.L⁻¹ BAP for subculture). Acclimatization was performed after third subculture. The apical shoot size of 0.5−1.0 cm is optimum for producing chrysanthemum-rooted cuttings. Meanwhile, MS medium for initiation stage, followed by MS + 2.5 mg.L⁻¹ GA₃ for subculture is the best culture medium composition for in vitro propagation. This explant and culture medium composition produced higher chlorophyll a, b, and a+b content, thereby resulting in higher plant, more leaves, larger stem diameter, and longer root length. This are recommended for chrysanthemum propagation, particularly in 'Jayanti Agrihorti'.

Purpose: The objective of the present study was to examine the impact of explant type and varying concentrations of 2,4-Dichlorophenoxyacetic acid and 6-Benzyladenine growth regulators on the San Pedro cactus callus morphological and biochemical characteristics. Research method: Four types of explants were used i.e. explants with areola, without areola, with truncated areola, and with central tissue. Additionally, five combinations of BA and 2,4-D, were tested (0 mg/L BA + 2 mg/L 2,4-D, 2 mg/L BA + 2 mg/L 2,4-D, 3 mg/L BA + 3 mg/L 2,4-D, 4 mg/L BA + 4 mg/L 2,4-D, 0 mg/L BA + 0 mg/L 2,4-D). Findings: The results indicated that callus formation induced in all treatments 6 days after inoculation. There were significant differences in growth parameters, including fresh weight, volume, moisture, tissue firmness, total phenols, total flavonoids and antioxidant activity of the callus (P < 0.01) and dry weight of callus (P < 0.05). Explants holding a segment of central tissue, yielded the least favorable results in most of experimental treatments, and the application of 2,4-D in the absence of BA had an inhibitory and toxic effect on the San Pedro cactus explants. Research limitations: No limitations were found. Originality/Value: Specifically, use of 2 mg/L BA + 2 mg/L 2,4-D and explants with areola resulted in callus with higher fresh weight, volume and total flavonoids, as well as good tissue integrity and firmness. The reported results are a valuable resource for future research related to cell tissue culture and the elicitation of secondary metabolites in Echinopsis spp.

Abstract Background Ovarian cancer is the first cause of death from gynecological malignancies mainly due to development of chemoresistance. Despite the emergence of PARP inhibitors, which have revolutionized the therapeutic management of some of these ovarian cancers, the 5-year overall survival rate remains around 45%. Therefore, it is crucial to develop new therapeutic strategies, to identify predictive biomarkers and to predict the response to treatments. In this context, functional assays based on patient-derived tumor models could constitute helpful and relevant tools for identifying efficient therapies or to guide clinical decision making. Method The OVAREX study is a single-center non-interventional study which aims at investigating the feasibility of establishing in vivo and ex vivo models and testing ex vivo models to predict clinical response of ovarian cancer patients. Patient-Derived Xenografts (PDX) will be established from tumor fragments engrafted subcutaneously into immunocompromised mice. Explants will be generated by slicing tumor tissues and Ascites-Derived Spheroids (ADS) will be isolated following filtration of ascites. Patient-derived tumor organoids (PDTO) will be established after dissociation of tumor tissues or ADS, cell embedding into extracellular matrix and culture in specific medium. Molecular and histological characterizations will be performed to compare tumor of origin and paired models. Response of ex vivo tumor-derived models to conventional chemotherapy and PARP inhibitors will be assessed and compared to results of companion diagnostic test and/or to the patient’s response to evaluate their predictive value. Discussion This clinical study aims at generating PDX and ex vivo models (PDTO, ADS, and explants) from tumors or ascites of ovarian cancer patients who will undergo surgical procedure or paracentesis. We aim at demonstrating the predictive value of ex vivo models for their potential use in routine clinical practice as part of precision medicine, as well as establishing a collection of relevant ovarian cancer models that will be useful for the evaluation of future innovative therapies. Trial registration The clinical trial has been validated by local research ethic committee on January 25th 2019 and registered at ClinicalTrials.gov with the identifier NCT03831230 on January 28th 2019, last amendment v4 accepted on July 18, 2023.

Melatonin is a pineal hormone that modulates the circadian system and exerts soporific and phase‐shifting effects. It is also involved in many other physiological processes, such as those implicated in cardiovascular, endocrine, immune, and metabolic functions. However, the role of melatonin in glucose metabolism remains contradictory, and its action on human adipose tissue (AT) explants has not been demonstrated. We aimed to assess whether melatonin (a pharmacological dose) influences insulin sensitivity in human AT. This will help better understand melatonin administration's effect on glucose metabolism. Abdominal AT (subcutaneous and visceral) biopsies were obtained from 19 participants with severe obesity (age: 42.84 ± 12.48 years; body mass index: 43.14 ± 8.26 kg/m2) who underwent a laparoscopic gastric bypass. AT biopsies were exposed to four different treatments: control (C), insulin alone (I) (10 nM), melatonin alone (M) (5000 pg/mL), and insulin plus melatonin combined (I + M). All four conditions were repeated in both subcutaneous and visceral AT, and all were performed in the morning at 8 a.m. (n = 19) and the evening at 8 p.m. (in a subsample of n = 12). We used western blot analysis to determine insulin signaling (using the pAKT/tAKT ratio). Furthermore, RNAseq analyses were performed to better understand the metabolic pathways involved in the effect of melatonin on insulin signaling. As expected, insulin treatment (I) increased the pAKT/tAKT ratio compared with control (p <.0001). Furthermore, the addition of melatonin (I + M) resulted in a decrease in insulin signaling as compared with insulin alone (I); this effect was significant only during the evening time (not in the morning time). Further, RNAseq analyses in visceral AT during the evening condition (at 8 p.m.) showed that melatonin resulted in a prompt transcriptome response (around 1 h after melatonin addition), particularly by downregulating the insulin signaling pathway. Our results show that melatonin reduces insulin sensitivity in human AT during the evening. These results may partly explain the previous studies showing a decrease in glucose tolerance after oral melatonin administration in the evening or when eating late when endogenous melatonin is present. [ABSTRACT FROM AUTHOR]

Capsicum annuum L. is extensively cultivated in subtropical and temperate regions globally, respectively, when grown in a medium with 8 holding significant economic importance. Despite the availability of genome sequences and editing tools, gene editing in peppers is limited by the lack of a stable regeneration and transformation method. This study assessed regeneration and transformation protocols in seven chili pepper varieties, including CM334, Zunla-1, Zhongjiao6 (ZJ6), 0818, 0819, 297, and 348, in order to enhance genetic improvement efforts. Several explants, media compositions, and hormonal combinations were systematically evaluated to optimize the in vitro regeneration process across different chili pepper varieties. The optimal concentrations for shoot formation, shoot elongation, and rooting in regeneration experiments were determined as 5 mg/L of 6-Benzylaminopurine (BAP) with 5 mg/L of silver nitrate (AgNO3), 0.5 mg/L of Gibberellic acid (GA3), and 1 mg/L of Indole-3-butyric acid (IBA), respectively. The highest regeneration rate of 41% was observed from CM334 cotyledon explants. Transformation optimization established 300 mg/L of cefotaxime for bacterial control, with a 72-h co-cultivation period at OD600 = 0.1. This study optimizes the protocols for chili pepper regeneration and transformation, thereby contributing to genetic improvement efforts. [ABSTRACT FROM AUTHOR]

Pacific oysters (Crassostrea or Magallana gigas) are one of the most economically important aquaculture species globally. Over the past two decades, ostreid herpesvirus (OsHV-1) has become a major pathogen of cultured Pacific oysters, resulting in widespread mortality with a global distribution. Experimental use of OsHV-1 is challenging for many reasons, including both complexity of host–pathogen dynamics and a lack of functioning model systems. The goal of this study was to improve the tools available for working with OsHV-1 in both whole animals and in tissue explants established from oysters maintained in controlled laboratory conditions. Tissue explants were taken from oysters originating from two different sources that have different levels of mortality in experimental OsHV-1 infections and were exposed to OsHV-1. A whole-animal infection experiment was run concurrently as a comparison. Quantitative PCR and electron microscopy were used to confirm that the explants were capable of replicating OsHV-1. Furthermore, the quantitative PCR results suggest that the source of the oysters was significant in determining the outcome of infection in the explants, supporting the validity of the explant model for OsHV-1 infection. This tissue explant approach for studying OsHV-1 allows for the control of confounding factors in the disease outcome that is not possible in whole-animal experiments, providing a new tool for the study of OsHV-1 in Pacific oysters. [ABSTRACT FROM AUTHOR]

Yam (Dioscorea sp.) is an important staple crop for millions of smallholder farmers in West Africa. In this study, we aimed to develop an efficient indirect plant regeneration system for three important yam genotypes "Kukrupa," "Mankrong Pona" and "Matches." The potential of leaf primordia, petioles and axillary bud explants for callus induction on Murashige and Skoog (MS) medium supplemented with different concentrations (0, 3, 5, 7 and 9 mg/l) of 2,4-dicholorophenoxy acetic acid (2,4-D) or picloram were tested. Callus induction was influenced by auxin concentration as well as explant. The highest callus induction was achieved using leaf primordia ("Kukrupa" and "Mankrong Pona") explants on 3 mg/l picloram or axillary buds ("Matches") on 5 mg/l picloram. Transfer of callus to 6-benzylaminoapurine (BAP) amended medium resulted in the formation of at least two shoots per callus clump. Plantlets were obtained on basic yam medium supplemented with 0.2 mg/l 1-naphthalene acetic acid (NAA), 3 mg/l kinetin and 0.5 mg/l BAP after four weeks. Post acclimatization plantlet survival was high (75 − 82.5%). The indirect plant regeneration protocol reported through this study helps in the improvement of important yam genotypes through tissue culture-oriented genome editing, in vitro mutagenesis and also for its mass propagation. [ABSTRACT FROM AUTHOR]

'Jayanti Agrihorti' is a superior chrysanthemum therefore rooted cuttings quality is required. In vitro propagation increases the rate of propagation and produces true-to-type plants. The research to obtain the best explants and culture medium composition that is capable of producing high-quality plants. The research was arranged in a randomized complete block design consisting of two factors. The first factor was the explant, including the apical shoot of 0.5 cm, 1.0 cm, and the nodal segment. The second factor was in vitro culture medium composition (CP), consisting of CP1 (Murashige and Skoog (MS) medium for initiation, followed by MS + 2.5 mg.L-1 gibberellic acid (GA3) for subculture), CP2 (MS + 0.25 mg.L-1 benzyl amino purine (BAP) for initiation, followed by MS for subculture), CP3 (MS + 0.25 mg.L-1 BAP for initiation, followed by MS + 2.5 mg.L-1 GA3 for subculture), and CP4 (MS + 0.5 mg.L-1 BAP for initiation, followed by MS + 0.25 mg.L-1 BAP for subculture). Acclimatization was performed after third subculture. The apical shoot size of 0.5-1.0 cm is optimum for producing chrysanthemum-rooted cuttings. Meanwhile, MS medium for initiation stage, followed by MS + 2.5 mg.L-1 GA3 for subculture is the best culture medium composition for in vitro propagation. This explant and culture medium composition produced higher chlorophyll a, b, and a+b content, thereby resulting in higher plant, more leaves, larger stem diameter, and longer root length. This are recommended for chrysanthemum propagation, particularly in 'Jayanti Agrihorti'. [ABSTRACT FROM AUTHOR]

Silybum marianum L. is considered one of the most extensively used medicinal plants worldwide due to its therapeutic benefits. While ultrasound waves were used to enhance the properties of numerous plant species. However, no study was investigated on applying ultrasonic waves to this particular plant. Therefore, the study amid to assess the impact of different exposure periods (0,10, 20, 25, 30, 35, and 40 minutes) of a frequency of 47.6 KHz on callus induction, protein content, and plant regeneration in S. marianum. The effect of ultrasonication was distinctive in accelerating callus induction of S. marianum, especially in short exposing periods (10 and 20 minutes). The percentage of callus formation reached 100%, 83.3% for cotyledons and stem respectively, when exposed to 20 minutes of ultrasound. Additionally, growth and total protein content were increased at 40 days and 80 days post- treatments. In contrast, long exposing periods (30, 35, and 40 minutes) had a negative impact on callus induction from all explants, as well as on callus growth and protein content. Moreover, ultrasonication stimulated one-step shoot regeneration during callus induction. The percentage of this phenomenon reached 100% for cotyledon at 10 and 20 minutes exposing period. This study confirmed the advantages of applying ultrasonic waves, particularly during shorter period to enhance the cultivation of S. marianum in vitro. [ABSTRACT FROM AUTHOR]

Plant growth regulators (PGRs) control signaling networks and developmental processes involved in plant responses to various biotic and abiotic stresses, making it crucial to study PGRs in vitro. The protocol for micropropagation of Solanum lycopersicum L., following callus induction and regeneration through explants such as internodal segments, leaves, and nodal segments, was established during the present study. Explants were inoculated on Murashige and Skoog (MS) medium supplemented with different plant growth regulators like BA (6-benzylaminopurine), 2,4-D (2,4-dichlorophenoxyacetic acid), BA + 2,4-D, IAA (Indoleacetic acid), IBA (Indolebutyric acid), and NAA (Naphthaleneacetic acid). It was observed that among all explants, the nodal segment showed maximum callus induction (83.33%) and multiplication (86.67%) at 0.25 mg/l of 2,4-D; the highest shoot number (3.33) at 0.5 mg/l of IAA + 0.5 mg/l of BA; the greatest shoot length (7.57 cm) at 0.75 mg/l of BA; root induction (80.95%), root number (21.88), and root length (10.62 cm) at 1 mg/l of IAA. Additionally, the maximum fresh weight (2.448 g) was observed at 0.5 mg/l of BA, while the highest dry weight (0.172 g) and dry matter content (14.25%) were noted at 1 mg/l of BA + 1 mg/l of 2,4-D for the internodal segment. Results of the present study revealed that among different explants, the best response was given by nodal segments, followed by internodal segments. Among the different PGRs, 2,4-D resulted in the highest callus induction and multiplication percentage. [ABSTRACT FROM AUTHOR]

Background: Ovarian cancer is the first cause of death from gynecological malignancies mainly due to development of chemoresistance. Despite the emergence of PARP inhibitors, which have revolutionized the therapeutic management of some of these ovarian cancers, the 5-year overall survival rate remains around 45%. Therefore, it is crucial to develop new therapeutic strategies, to identify predictive biomarkers and to predict the response to treatments. In this context, functional assays based on patient-derived tumor models could constitute helpful and relevant tools for identifying efficient therapies or to guide clinical decision making. Method: The OVAREX study is a single-center non-interventional study which aims at investigating the feasibility of establishing in vivo and ex vivo models and testing ex vivo models to predict clinical response of ovarian cancer patients. Patient-Derived Xenografts (PDX) will be established from tumor fragments engrafted subcutaneously into immunocompromised mice. Explants will be generated by slicing tumor tissues and Ascites-Derived Spheroids (ADS) will be isolated following filtration of ascites. Patient-derived tumor organoids (PDTO) will be established after dissociation of tumor tissues or ADS, cell embedding into extracellular matrix and culture in specific medium. Molecular and histological characterizations will be performed to compare tumor of origin and paired models. Response of ex vivo tumor-derived models to conventional chemotherapy and PARP inhibitors will be assessed and compared to results of companion diagnostic test and/or to the patient's response to evaluate their predictive value. Discussion: This clinical study aims at generating PDX and ex vivo models (PDTO, ADS, and explants) from tumors or ascites of ovarian cancer patients who will undergo surgical procedure or paracentesis. We aim at demonstrating the predictive value of ex vivo models for their potential use in routine clinical practice as part of precision medicine, as well as establishing a collection of relevant ovarian cancer models that will be useful for the evaluation of future innovative therapies. Trial registration: The clinical trial has been validated by local research ethic committee on January 25th 2019 and registered at ClinicalTrials.gov with the identifier NCT03831230 on January 28th 2019, last amendment v4 accepted on July 18, 2023. [ABSTRACT FROM AUTHOR]

Purpose: The objective of the present study was to examine the impact of explant type and varying concentrations of 2,4-Dichlorophenoxyacetic acid and 6-Benzyladenine growth regulators on the San Pedro cactus callus morphological and biochemical characteristics. Research method: Four types of explants were used i.e. explants with areola, without areola, with truncated areola, and with central tissue. Additionally, five combinations of BA and 2,4-D, were tested (0 mg/L BA + 2 mg/L 2,4-D, 2 mg/L BA + 2 mg/L 2,4-D, 3 mg/L BA + 3 mg/L 2,4-D, 4 mg/L BA + 4 mg/L 2,4-D, 0 mg/L BA + 0 mg/L 2,4-D). Findings: The results indicated that callus formation induced in all treatments 6 days after inoculation. There were significant differences in growth parameters, including fresh weight, volume, moisture, tissue firmness, total phenols, total flavonoids and antioxidant activity of the callus (P < 0.01) and dry weight of callus (P < 0.05). Explants holding a segment of central tissue, yielded the least favorable results in most of experimental treatments, and the application of 2,4-D in the absence of BA had an inhibitory and toxic effect on the San Pedro cactus explants. Research limitations: No limitations were found. Originality/Value: Specifically, use of 2 mg/L BA + 2 mg/L 2,4-D and explants with areola resulted in callus with higher fresh weight, volume and total flavonoids, as well as good tissue integrity and firmness. The reported results are a valuable resource for future research related to cell tissue culture and the elicitation of secondary metabolites in Echinopsis spp. [ABSTRACT FROM AUTHOR]

The cultivation of Vanda orchids in Indonesia faces challenges due to their slow growth, leading to a reliance on importing varieties from foreign countries. Our primary objectives were understanding the relationship between morphological characters and in vitro propagation success and establishing mass in vitro propagation technology from initiation to maturation stages. Additionally, we explored the influence of genotypes, explant types, and media during the initiation stage, alongside evaluating the media's role in the proliferation, regeneration, and maturation of plantlets. A well-defined protocol was developed for the culture induction, multiplication, rejuvenation, and development of Vanda orchids. Established correlations highlight the correlation between floral structure colors and the effectiveness of explant initiation. The optimal medium for culture initiation was Murashige & Skoog (MS) with the addition of 0.75 mg/l thidiazuron (TDZ) and 0.25 mg/l 6-benzyl amino purine (BAP), particularly effective by using rachis explants with the best response found in V. ‘Nilareta Agrihorti’, which exhibited organogenic callus and adventitious shoots within 58 days. During the proliferation stage, culturing 0.5 cm clumps on MS medium with 1 and 2 mg/l BAP resulted in a substantial 3.30 and 2.83 times increasing in clump weight, respectively, and a multiplication rate of 4.30 and 3.83, respectively. MS was added with 0.3 mg/l meta-topoline for shoot regeneration, achieving an average shoot growth percentage of up to 89% during the third subculture period. Half-strength MS medium was combined with 100 g/l banana extract, demonstrating superior plantlet performance during maturation. These findings represent a significant advancement in Vanda hybrid propagation technology, holding promise for cultivating and commercializing these unique orchid varieties in Indonesia and international market.Key message: We examines genotype-driven in vitro culture responses in Vanda, revealing correlations between floral traits and explant performances. Optimization of media, and explants enhances initiation, proliferation, regeneration, and plantlet maturation success. [ABSTRACT FROM AUTHOR]

During mouse development, presomitic mesoderm cells synchronize Wnt and Notch oscillations, creating sequential phase waves that pattern somites. Traditional somitogenesis models attribute phase waves to a global modulation of the oscillation frequency. However, increasing evidence suggests that they could arise in a self-organizing manner. Here, we introduce the Sevilletor, a novel reaction-diffusion systemthat serves as a framework to compare different somitogenesis patterning hypotheses. Using this framework, we propose the Clock and Wavefront Self-Organizing model that considers an excitable selforganizing region where phase waves form independent of global frequency gradients. The model recapitulates the change in relative phase of Wnt and Notch observed during mouse somitogenesis and provides a theoretical basis for understanding the excitability of mouse presomitic mesoderm cells in vitro. [ABSTRACT FROM AUTHOR]

The existing study held in the Plant Tissue Culture Laboratory, University of Tikrit, Iraq, sought to increase the production of some secondary metabolite compounds in tissue cultures of Chrysanthemum hortorum Hort. cv. 'Dwarf White.' Inducing callus by culturing the bases of young leaf explants on the MS medium received supplementation of different concentrations of Benzyl adenine (BA) (0, 1.5, and 2 mg L-1) and Indole-3-acetic acid (IAA) (0.0, 0.5, and 1.5 mg L-1). Tyrosine addition at different concentrations (0, 30, 60, and 80 mg L-1) and sucrose at 30, 60, 80, and 100 g L-1 concentrations ensued. In addition, fructose and glucose applications at 90.0, 60.0, 30.0, and 120.0 g L-1 transpired for the callus growth. The combination of 2.0 mg L-1 BA + 1.5 mg L-1 IAA gave the highest average fresh and dry weights of callus, reaching 1.8 and 5.72 mg, respectively. The best treatment was 60 g L-1, which ecorded the maximum pyrethrin concentration, amounting to 2.134 µg/ml DW. The treatment of 90 g L-1 fructose + 60 mg L-1 tyrosine was more effective in increasing pyrethrin production in the callus, reaching 3.175 µg/ml DW. The treatment of 90 g L-1 glucose + 60 mg L-1 tyrosine was recorded with the utmost pyrethrin concentration, reaching 3.346 µg/ml DW. The treatment of 90 g L-1 glucose + 80 mg L-1 tyrosine provided 2.826 µg/ml DW of pyrethrin. [ABSTRACT FROM AUTHOR]

The cultivation of raja banana is widespread in Indonesia. The process of increasing banana propagation through in vitro culture encounters a specific issue, namely the occurrence of browning during the initiation stage, which hinders the regeneration process. The objective of this study is to determine the effectiveness of antioxidants on the in vitro regeneration of raja banana to prevent browning and enhance embryo development. The study was conducted using a completely randomized design with the treatment of the antioxidant compounds ascorbic acid (AS) and melatonin (MN). The treatments consisted of control (without antioxidants), ascorbic acid (100 mg L-1, 150 mg L-1, and 200 mg L-1) and melatonin (10 mg L-1, 12 mg L-1, and 14 mg L-1). The results showed that ascorbic acid and melatonin treatment had no significant effect on the percentage of viable explants and the level of browning intensity, but provided significant results on the regeneration process. The application of melatonin at 14 mg L-1 significantly increased callus regeneration. Furthermore, at a concentration of 12 mg L-1 showed the highest callus percentage value compared to other treatments. The melatonin 12 mg L-1 treatment showed the earliest scutellar embryo formation, whereas ascorbic acid at a concentration of 100 mg L-1 resulted in the most optimal regeneration of globular and scutellar embryos. The highest concentrations of ascorbic acid and melatonin inhibit the formation of coleoptilar embryos

Type I collagen is a predominant fibrous protein that makes up the extracellular matrix. Collagen enhances cell attachment and is commonly used in three-dimensional culture systems, to mimic the native extracellular environment, for primary sensory neurons such as dorsal root ganglia (DRG). However, the effects of collagen concentration on adult rat DRG neurite growth have not been assessed in a physiologically relevant, three-dimensional culture. This study focuses on the effects of type I collagen used in a methacrylated hyaluronic acid (MAHA)-laminin-collagen gel (triple gel) on primary adult rat DRG explants in vitro. DRGs were cultured in triple gels, and the neurite lengths and number of support cells were quantified. Increased collagen concentration significantly reduced neurite length but did not affect support cell counts. Mechanical properties, fiber diameter, diffusivity, and mesh size of the triple gels with varying collagen concentration were characterized to further understand the effects of type I collagen on hydrogel property that may affect adult rat DRG explants. Gel stiffness significantly increased as collagen concentration increased and is correlated to DRG neurite length. Collagen concentration also significantly impacted fiber diameter but there was no correlation with DRG neurite length. Increasing collagen concentration had no significant effect on mesh size and diffusivity of the hydrogel. These data suggest that increasing type I collagen minimizes adult rat DRG explant growth in vitro while raising gel stiffness. This knowledge can help develop more robust 3D culture platforms to study sensory neuron growth and design biomaterials for nerve regeneration applications. [ABSTRACT FROM AUTHOR]

Surface sterilization is crucial in preparing viable and uncontaminated explants for tissue culture. Utilizing the right sterilizing agent in this procedure can efficiently eliminate the majority of surface contaminants. The present study aimed to establish the best sterilization procedure for clonal apple rootstock MM106 using shoot tips and nodal segments. Two important sterilizing agents, mercuric chloride (ranging from 0.05% to 0.3%) for various durations (2 to 6 minutes) and sodium hypochlorite (ranging from 3.0% to 12%) for different durations (10 to 30 minutes), were employed. The results indicated that the maximum aseptic conditions (80.00% and 80.67%) and survival rates of the cultures (60.66% and 67.33%) for shoot tips and nodal segments, respectively, were achieved through surface sterilization with 0.1% mercuric chloride for 4 minutes, specifically during April, in comparison to March and May of 2021 due to low phenol content and high meristematic activity. The results yielded valuable insights for efficiently propagating clonal apple rootstocks on a local scale, and they have the potential to offer guidance for establishing commercial facilities dedicated to producing these specific clonal apple rootstocks and varieties. [ABSTRACT FROM AUTHOR]

Degenerative meniscus lesions have been associated with both osteoarthritis etiology and its progression. We, therefore, sought to establish a human meniscus ex vivo model to study the meniscal response to cytokine treatment using a proteomics approach. Lateral menisci were obtained from five knee‐healthy donors. The meniscal body was cut into vertical slices and further divided into an inner (avascular) and outer region. Explants were either left untreated (controls) or stimulated with cytokines. Medium changes were conducted every 3 days up to Day 21 and liquid chromatography–mass spectrometry was performed at all the time points for the identification and quantification of proteins. Mixed‐effect linear regression models were used for statistical analysis to estimate the effect of treatments versus control on protein abundance. Treatment by IL1ß increased release of cytokines such as interleukins, chemokines, and matrix metalloproteinases but a limited catabolic effect in healthy human menisci explants. Further, we observed an increased release of matrix proteins (collagens, integrins, prolargin, tenascin) in response to oncostatin M (OSM) + tumor necrosis factor (TNF) and TNF+interleukin‐6 (IL6) + sIL6R treatments, and analysis of semitryptic peptides provided additional evidence of increased catabolic effects in response to these treatments. The induced activation of catabolic processes may play a role in osteoarthritis development. [ABSTRACT FROM AUTHOR]

Indications for implant removal after fracture healing are still under debate in both human and veterinary medicine. Although hardware removal is a common procedure, it should not be undertaken lightly. Intra and post-operative complications are common and a thorough evaluation of the risks and benefits should be performed. This review aimed to collect and summarise published data on the indications for implant removal in small animals, compare the collected data with human and equine medicine, and investigate the existence of guidelines for this purpose. There is no international consensual agreement for implant removal after fracture healing, neither in small animals nor in human orthopaedics. Decision-making processes are still controversial in some scenarios, thus clear evidence-based protocols for implant removal are needed.

[Objectives] This study was conducted to increase the reproduction coefficient of Japanese honeysuckle (Lonicera japonica) to keep the character of optimal benign. [Methods] The young leaves of medicinal Japanese honeysuckle were selected as explants, and MS was used as the basic culture medium. Suitable culture concentrations and conditions were screened through different concentration gradients of growth regulators and cytokinin. [Results] As the concentration of 6-BA in the culture medium increased, the browning rate increased, and the browning phenomenon occurred earlier. On the contrary, a lower concentration of6-BA was suitable for the differentiation and growth of young leaves, and the browning response was slow. However, if the cultivation time was too long and the materials were not transferred in a timely manner, browning would also occur. The optimal combination of levels was obtained through a 3 X 3 orthogonal experiment (three parallel groups for each of 6-BA and NAA). The culture conditions included a constant temperature of 26 O and light intensity of 1 200 lx. The optimal medium for inducing callus proliferation was MS P 6-BA 0. 5 mg/L P NAA 0. 5 mg/L; and the optimal medium for inducing bud differentiation was MS P 6-BA 1. 0 mg/L P NAA 0.1 mg/L. [Conclusions] This study provides a theoretical basis for accelerating the development of the honeysuckle industry. [ABSTRACT FROM AUTHOR]

Breast cancer is one of the main causes of death worldwide. Lately, there is great interest in developing methods that assess individual sensitivity and/or resistance of tumors to antineoplastics to provide personalized therapy for patients. In this study we used organotypic culture of human breast tumor slices to predict the experimental effect of antineoplastics on the viability of tumoral tissue. Samples of breast tumor were taken from 27 patients with clinically advanced breast cancer; slices were obtained and incubated separately for 48 h with paclitaxel, docetaxel, epirubicin, 5-fluorouracil, cyclophosphamide, and cell culture media (control). We determined an experimental tumor sensitivity/resistance (S/R) profile by evaluating tissue viability using the Alamar Blue® metabolic test, and by structural viability (histopathological analyses, necrosis, and inflammation). These parameters were related to immunohistochemical expression of the estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. The predominant histological type found was infiltrating ductal carcinoma (85.2%), followed by lobular carcinoma (7.4%) and mixed carcinoma (7.4%). Experimental drug resistance was related to positive hormone receptor status in 83% of samples treated with cyclophosphamide (p = 0.027). Results suggest that the tumor S/R profile can help to predict personalized therapy or optimize chemotherapeutic treatments in breast cancer. [ABSTRACT FROM AUTHOR]

Lemon (Citrus limon) cultivars (Desi and Chinese) were subjected to gamma radiations for mutation induction. The current investigation was performed with the objective of exploring the effect of gamma irradiation on morphological characteristics of lemon and for development of lemon mutants for the sake of crop improvement. Seeds and bud wood of two lemon cultivars were exposed to various gamma ray concentrations (i.e 20,50,80,110 and 140 Gy). The irradiated buds were then budded on a rootstock (rough lemon) at standard growth stage of the root stock and seeds were sown in sterilized growing media in order to get disease free plants with maximum germination. The experimentation was carried out according to Randomized Complete Block Design (RCBD). The recorded observations were arranged and analyzed using statistical software Statistix 8.1 for employing statistical techniques. The research findings of the current investigation showed that there was significant variation in morphological variables among the irradiated and control plants of both cultivars. With the increase in radiation intensity, there was marked reduction in growth variables of both lemon varieties. [ABSTRACT FROM AUTHOR]

Meibomian gland dysfunction (MGD) is a group of disorders linked by functional abnormalities of the meibomian glands. Current studies on MGD pathogenesis focus on meibomian gland cells, providing information on a single cell's response to experimental manipulation, and do not maintain the architecture of an intact meibomian gland acinus and the acinar epithelial cells' secretion state in vivo. In this study, rat meibomian gland explants were cultured by a Transwell chamber-assisted method under an air-liquid interface (airlift) in vitro for 96 h. Analyses for tissue viability, histology, biomarker expression, and lipid accumulation were performed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and TUNEL assays, hematoxylin and eosin (H&E) staining, immunofluorescence, Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), transmission electron microscopy (TEM), and western blotting (WB). MTT, TUNEL, and H&E staining indicated better tissue viability and morphology than the submerged conditions used in previous studies. Levels of MGD biomarkers, including keratin 1 (KRT1) and 14 (KRT14) and peroxisome proliferator-activated receptor-gamma (PPAR-γ), along with oxidative stress markers, including reactive oxygen species, malondialdehyde, and 4-hydroxy-2-nonenal, gradually increased over culture time. The MGD pathophysiological changes and biomarker expression of meibomian gland explants cultured under airlift conditions were similar to those reported by previous studies, indicating that abnormal acinar cell differentiation and glandular epithelial cell hyperkeratosis may contribute to obstructive MGD occurrence. [ABSTRACT FROM AUTHOR]

The CRISPR/Cas9 system is extensively used for plant gene editing. This study developed an efficient CRISPR/Cas9 system for Chinese kale using multiple sgRNAs and two promoters to create various CRISPR/Cas9 vectors. These vectors targeted BoaZDS and BoaCRTISO in Chinese kale protoplasts and cotyledons. Transient transformation of Chinese kale protoplasts was assessed for editing efficiency at three BoaZDS sites. Notably, sgRNA: Z2 achieved the highest efficiency (90%). Efficiency reached 100% when two sgRNAs targeted BoaZDS with a deletion of a large fragment (576 bp) between them. However, simultaneous targeting of BoaZDS and BoaCRTISO yielded lower efficiency. Transformation of cotyledons led to Chinese kale mutants with albino phenotypes for boazds mutants and orange-mottled phenotypes for boacrtiso mutants. The mutation efficiency of 35S-CRISPR/Cas9 (92.59%) exceeded YAO-CRISPR/Cas9 (70.97%) in protoplasts, and YAO-CRISPR/Cas9 (96.49%) surpassed 35S-CRISPR/Cas9 (58%) in cotyledons. These findings introduce a strategy for enhancing CRISPR/Cas9 editing efficiency in Chinese kale. [ABSTRACT FROM AUTHOR]

Abstract In 2019 a low pathogenic H3N1 avian influenza virus (AIV) caused an outbreak in Belgian poultry farms, characterized by an unusually high mortality in chickens. Influenza A viruses of the H1 and H3 subtype can infect pigs and become established in swine populations. Therefore, the H3N1 epizootic raised concern about AIV transmission to pigs and from pigs to humans. Here, we assessed the replication efficiency of this virus in explants of the porcine respiratory tract and in pigs, using virus titration and/or RT-qPCR. We also examined transmission from directly, intranasally inoculated pigs to contact pigs. The H3N1 AIV replicated to moderate titers in explants of the bronchioles and lungs, but not in the nasal mucosa or trachea. In the pig infection study, infectious virus was only detected in a few lung samples collected between 1 and 3 days post-inoculation. Virus titers were between 1.7 and 4.8 log10 TCID50. In line with the ex vivo experiment, no virus was isolated from the upper respiratory tract of pigs. In the transmission experiment, we could not detect virus transmission from directly inoculated to contact pigs. An increase in serum antibody titers was observed only in the inoculated pigs. We conclude that the porcine respiratory tract tissue explants can be a useful tool to assess the replication efficiency of AIVs in pigs. The H3N1 AIV examined here is unlikely to pose a risk to swine populations. However, continuous risk assessment studies of emerging AIVs in pigs are necessary, since different virus strains will have different genotypic and phenotypic traits.

A study was carried out to standardize the sterilization protocol for different explants (terminal stem scale, immature flower bud and tepal segment) and to select the suitable explant for the direct organogenesis of tuberose cv. Arka Vaibhav. The highest survival per cent (100) and uncontaminated cultures (0.00) of terminal stem scale explant was observed in pre-treatment with overnight soaking of terminal stem scale in the solution comprising carbendazim (0.1%), chlorothalonil (0.05%) and myristyl trimethyl ammonium bromide (cetrimide) (0.05%) and subsequently surface sterilization with 70% ethanol (1 min), 4% sodium hypochlorite (10 min) followed by 0.1% HgCl2 (15 min). The explant immature flower bud recorded the highest survival per cent (100) and maximum aseptic cultures in the treatment T1 comprised of 1.0 drop Tween-20 + 70% ethanol (30 sec) and 1% sodium hypochlorite (3 min). Pre-treatment of tepal segment explant in 0.1% carbendazim (30 min) solution followed by surface sterilization with combination of 1.0 drop Tween-20 + 70% ethanol (30 sec) followed by 1% sodium hypochlorite (3 min) registered 91.66% of survival with the minimum contamination (10%) in the treatment. Among the three explants used, the terminal stem scale was found suitable for direct organogenesis with early greenness (5.72 days) and highly responsive to shoot induction (100%) in MS medium supplemented with 4 mg/L BAP + 0.1mg/L IAA. Other two explants viz., immature flower bud and tepal segment failed to respond for direct organogenesis by shoot induction instead produced profuse callus. [ABSTRACT FROM AUTHOR]

Callus induction was attempted from the four explants viz. root, cotyledon, epicotyl, and leaf segments excised from in vitro raised seedlings of C. jambhiri. Among various MS media supplementations with growth regulators and carbohydrates, the maximum (95.50%) and the earliest (8.30 days) callogenesis was obtained in epicotyl segments, when cultured on MS medium supplemented with NAA (10.0 mgl-1) + BAP (1.0 mgl-1) + sucrose (8%). The modified MS (macro and micro-nutrients reduced to half) fortified with BAP (5.0 mgl-1) + GA3 (3.0 mgl-1) recorded maximum shoot regeneration (43.10%) from callus, with an average of 5.30 shoots per callus after 35.50 days of culturing. However, prolonged exposure to GA3 resulted in thin elongated shoots and leaves. The age of the callus substantially influenced the plant regeneration frequency. The potency of the callus to regenerate decreased significantly with an increase in the age of the callus. Shoot regeneration was recorded maximum (43.43%) in 60 days old calli, followed by 90 days old (30.48%) calli, whereas it was minimum (10.46%) in 150 days old calli. The maximum (79.50%) shoot proliferation was recorded in MS medium supplemented with BAP (1.0 mgl-1) + Kin (0.5 mgl-1) with an average of 5.06 shoots per culture. The MS medium fortified with NAA (1.0 mgl-1) + IBA (1.0 mgl-1) induced maximum (77.33%) rooting, with an average of 3.19 roots per shoot after 13.4 days of culturing. Rooted plants were hardened and survived the best (83.6%) on the potting mixture consisting of cocopeat + vermiculite + perlite (2:1:1). [ABSTRACT FROM AUTHOR]

An increased dependency on plant-based resources for food, shelter, and medicinal usage has increased their sustainable and unsustainable exploitation. To use this resource sustainably, plant tissue culture (PTC) is one important technology. Among different PTC techniques, thin cell layer (TCL) technology is a relatively simple and easily adaptable technique for in vitro cultures of plants. This technique uses small explants about 0.5–2 mm in thickness excised from different plant organs. It has been successfully used in the large-scale propagation of vegetables, legumes, and plants with medicinal benefits. TCL technology has proven to be effective in stimulating various organogenic responses when combined with various new methods such as nanotechnology or microtome-based explantation, especially in tree species. It is considered an important tool in plant biotechnology. Although the morphogenetic response per explant is usually higher in conventional explants, the appropriate use of plant growth regulators and geometric factors in TCL has the potential to make it more efficient and beneficial. This article provides an overview of the concept of TCL as applied to different plant species, particularly trees, since there are few, if any, summaries of TCL technology, especially in trees. This review will certainly revitalize this important technology so that it can be used effectively for successful mass propagation in the field of plant tissue culture. [ABSTRACT FROM AUTHOR]

Introduction The objective of the study was to characterize the baseline intra-individual and inter-individual variability of immune cell subsets within abdominoplasty skin specimens. Material and methods Abdominoplasty biopsies were taken from 5 patients and analysed using the Vectra 3 automated quantitative pathology imaging system with inForm software. Results Adjacent skin regions demonstrated intra-patient variability in immune subset counts ranging from 1- to 5-fold. Inter-variability between patients was approximately 2- to 7-fold for most subsets, except for HLA-DR+ antigen presenting cells, which varied 19-fold. Conclusions Our data highlight the importance of including multiple patients and multiple patient samples when designing dermatological studies that utilise abdominoplasty skin.

Background: Contamination of tissue culturemediumis acommonproblem that causes the loss of someor all cultures. Despite all measures taken in tissue culture laboratories to prevent culture contamination, bacterial and fungi contamination is still a significant problem under in vitro culture. Achieving a suitable method for controlling the contaminants in the in vitro culture medium will be a considerable development in the micropropagation of dates. Objectives: We aimed to investigate the effects of different aqueous and ethanolic extracts of Rhazya stricta on the disinfection of dates under in vitro culture. Methods: A factorial test was performed based on a completely randomized design with three replications. The study included three factors: plant organs (leaves and roots), solvent type (ethanolic and aqueous), and extract concentration (0, 25, 50, and 75 mg/mL). Results: The lowest contamination (38.33%) was seen in the treatment with ethanolic root extract (50 ppm). In comparison, the control and the treatment containing aqueous leaf extract (with concentrations of 0and 25ppm)resulted in the highest contamination rate (100%) in tissue culture conditions. Conclusions: The alcoholic extract of medicinal plants can be used to disinfect the explants and the substrate. [ABSTRACT FROM AUTHOR]

Orchids, with their astonishingly stunning flowers, dominate the international floricultural market. They are considered prized assets for commercial applications in pharmaceutical and floricultural industries as they possess high therapeutic properties and superior ornamental values. The alarming depletion of orchid resources due to excessive unregulated commercial collection and mass habitat destruction makes orchid conservation measures an extreme priority. Conventional propagation methods cannot produce adequate number of orchids, which would meet the requirement of these ornamental plants for commercial and conservational purposes. In vitro orchid propagation using semi-solid media offers an outstanding prospect of rapidly producing quality plants on a large scale. However, the semi-solid (SS) system has shortcomings with low multiplication rates and high production costs. Orchid micropropagation using a temporary immersion system (TIS) overcomes the limitations of the SS system by reducing production costs and making scaleup and full automation possible for mass plant production. The current review highlights different aspects of in vitro orchid propagation using SS and TIS and their benefits and drawbacks on rapid plant generation. [ABSTRACT FROM AUTHOR]

Abstract Background Reactive oxygen species (ROS) and calcium ions (Ca2+) are representative signals of plant wound responses. Wounding triggers cell fate transition in detached plant tissues and induces de novo root organogenesis. While the hormonal regulation of root organogenesis has been widely studied, the role of early wound signals including ROS and Ca2+ remains largely unknown. Results We identified that ROS and Ca2+ are required for de novo root organogenesis, but have different functions in Arabidopsis explants. The inhibition of the ROS and Ca2+ signals delayed root development in detached leaves. Examination of the auxin signaling pathways indicated that ROS and Ca2+ did not affect auxin biosynthesis and transport in explants. Additionally, the expression of key genes related to auxin signals during root organogenesis was not significantly affected by the inhibition of ROS and Ca2+ signals. The addition of auxin partially restored the suppression of root development by the ROS inhibitor; however, auxin supplementation did not affect root organogenesis in Ca2+-depleted explants. Conclusions Our results indicate that, while both ROS and Ca2+ are key molecules, at least in part of the auxin signals acts downstream of ROS signaling, and Ca2+ acts downstream of auxin during de novo root organogenesis in leaf explants.