Efficient Induction Research Articles

Inline induction heating for high-pressure fuel-based testing applications

Induction heating offers numerous advantages over conventional methods for heating high-pressure fluids in hydraulic testing applications. It simplifies complex processes, mitigates the risks associated with high-pressure environments, and enhances overall safety. The design of an efficient induction heating system requires a clear understanding of its key components, including the resonant tank circuit, impedance matching and isolation stage, high-frequency inverter, driver circuitry, and controller. The configuration of the resonant circuit plays a crucial role in device ratings and control methods. To meet the demanding requirements of switching devices like IGBTs, the driver circuit must ensure high immunity to interferences and failure. Effective system design allows the heating circuit to operate close to resonant switching, minimizing switching losses and cooling requirements. This paper presents the design and development of a 4-kW induction heater specifically tailored for inline heating of aviation fuel under high pressure. The system aims to achieve a precise temperature profile for testing line-replaceable units (LRUs) used in aircraft. The design methodology encompasses the selection of the resonant tank circuit configuration, impedance matching and isolation stage, high-frequency inverter, driver circuitry, and control strategies. Through extensive simulations in LTspice and experimental measurements, power losses in different components of the power conversion circuits are assessed, and overall system efficiency is evaluated. The results demonstrate the proposed induction heating system’s remarkable energy efficiency, reduced switching losses, and improved reliability. Furthermore, a comprehensive comparison with existing designs and industry norms is presented, highlighting the distinct advantages of the proposed system in terms of cost-effectiveness, energy efficiency, material utilization, and processing time. This research contributes valuable insights into the design and optimization of induction heating systems for high-pressure fluid heating applications, providing a practical and efficient solution for the aviation industry.

Various effects of repeated rifampin dosing on coproporphyrin levels in humans.

In recent years, the identification of endogenous substrates as biomarkers became an uprising topic. Particularly coproporphyrins (CPs), byproducts of heme biosynthesis, are intensely investigated as biomarkers for predicting interactions with the organic anion transporting polypeptide (OATP) 1B transporters. In the context of drug-drug interactions, several preclinical and clinical studies assessed the effect of the OATP1B-index inhibitor rifampin on CPI levels. However, rifampin is not only a "perpetrator" drug of transporters but is also known for its interaction with the nuclear receptor pregnane X receptor (PXR) leading to the efficient induction of PXR-target genes. These include hemoproteins like cytochrome P450 enzymes but also the δ-aminolevulinate synthase 1, which is the rate-limiting enzyme in heme biosynthesis. In this study, we showed that quantification of CPs in clinical serum samples was possible after long-term storage at -20°C. We quantified CPI, CPIII, and heme levels in clinical serum samples (at selected timepoints) that originated from a trial investigating the interaction potential of repeated rifampin administration in 12 healthy participants. In samples collected at the assumed time to maximum concentration of rifampin, higher CP levels were observed compared to baseline. Increased levels persisted even 14 h after discontinuation of rifampin. No impact on heme serum levels was observed. We found a correlation between CP isomers at baseline and at 14 h after rifampin intake. In summary, we show that multiple doses of rifampin affect CP levels. However, besides inhibition of hepatic OATP function there is evidence for an interaction with CP levels beyond the transporter level.

Improved compact growth habit of Viola × wittrockiana through Rhizobium rhizogenes transformation

Compactness is an important breeding goal in the improvement of garden pansy varieties (Viola × wittrockiana Gams.). The desired compact stature is primarily obtained through the use of chemical growth retardants. Recently, an alternative strategy for creating compact ornamentals was proposed based on transformation by wild type rhizogenic agrobacteria and the regeneration of transformed hairy roots. Here, we report on the successful transformation and creation of compact garden pansy lines. The most efficient hairy root induction was obtained by Rhizobium rhizogenes strain ATCC15834. Shoot regeneration from hairy roots was obtained using 3 mg L−1 2iP and 0.3 mg L−1 2,4-D. The presence of R. rhizogenes T-DNA genes was confirmed in the regenerated lines. These lines displayed a varying degrees of dwarfed growth. Most notably the shoot length was reduced up to 49%. Other phenotypic traits that were induced include reduced peduncle length and smaller, wrinkled flowers. Several of these Ri lines with promising compactness have been selected for further breeding towards compact garden pansy varieties that are less dependent on chemical growth regulation.Graphical abstract

Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray Poplar.

A series of tissue culture regeneration protocols were conducted on gray poplar (P. tremula × P. alba) to select the most efficient callus induction medium, adventitious shoot differentiation medium, shoot elongation medium and rooting medium, which laid the foundation for the optimization of genetic transformation technology for gray poplar. The results showed that the Woody Plant Medium (WPM) supplemented with 0.10 mg L-1 kinetin (KT) and 1.00 mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D) was the most suitable medium for callus induction. The callus induction rates of different tissues were greater than 85.7%. The optimal adventitious shoot differentiation medium was the WPM supplemented with 0.02 mg L-1 thidiazuron (TDZ), and the adventitious shoot differentiation rates of young tissues were 22.2-41.9%. The optimal direct differentiation medium was the Murashige and Skoog (MS) medium supplemented with 0.20 mg L-1 6-benzylaminopurine (6-BA), 0.10 mg L-1 indole butyric acid (IBA) and 0.001 mg L-1 TDZ, and the differentiation rate of adventitious shoots was greater than 94%. The best shoot elongation medium for adventitious shoots was the MS medium with 0.10 mg L-1 naphthylacetic acid (NAA). After 45 days of cultivation in the MS medium with 0.10 mg L-1 NAA, the average plant height was 1.8 cm, and the average number of elongated adventitious shoots was 11 per explant. The 1/2 MS medium with 0.10 mg L-1 NAA showed the best performance for rooting, and later, shoot growth. The direct shoot induction pathway can induce adventitious shoots much faster than the indirect adventitious shoot induction pathway can, and the time cost via the direct adventitious shoot induction pathway can be shortened by 2-6 weeks compared to that of the indirect shoot induction pathway.

Generation of Antibodies Selectively Recognizing Epitopes in a Formaldehyde-Fixed Cell-Surface Antigen Using Virus-like Particle Display and Hybridoma Technology.

Efficient induction of target-specific antibodies can be elicited upon immunization with highly immunogenic virus-like particles (VLPs) decorated with desired membrane-anchored target antigens (Ags). However, for example, for diagnostic purposes, monoclonal antibodies (mAbs) are required to enable the histological examination of formaldehyde-fixed paraffin-embedded (FFPE) biopsy tissue samples. Aiming at the generation of FFPE-antigen-specific mAbs and as a proof of concept (POC), we first established a simplified protocol using only formaldehyde and 90 °C heat fixation (FF90) of cells expressing the target Ag nerve growth factor receptor (NGFR). The FF90 procedure was validated using flow cytometric analysis and two mAbs recognizing either the native and FFPE-Ag or exclusively the native Ag. C-terminally truncated NGFR (trNGFR)-displaying native and FF90-treated VLPs derived from HIV-1 did not reveal distinctive changes in particle morphology using transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis. Mice were subsequently repetitively immunized with trNGFR-decorated FF90-VLPs and hybridoma technology was used to establish mAb-producing cell clones. In multiple screening rounds, nine cell clones were identified producing mAbs distinctively recognizing epitopes in FF90- and FFPE-NGFR. This POC of a new methodology should foster the future generation of mAbs selectively targeting FFPE-fixed cell-surface Ags.

Engineering Critical Residues of SOX9 Discovers a Variant with Potent Capacity to Induce Chondrocytes.

Articular cartilage plays vital roles as a friction minimizer and shock absorber during joint movement but has a poor capacity to self-repair when damaged through trauma or disease. Cartilage tissue engineering is an innovative technique for cartilage regeneration, yet its therapeutic application requires chondrocytes in large numbers. Direct reprogramming of somatic cells to chondrocytes by expressing SOX9, KLF4, and c-MYC offers a promising option to generate chondrocytes in sufficient numbers; however, the low efficiency of the reprogramming system warrants further improvement. Here we referred to structural and functional features of SOX9 and performed alanine-scanning mutagenesis of functionally critical residues in the HMG box and at putative posttranslational modification (PTM) sites. We discovered that a SOX9 variant H131A/K398A, doubly mutated in the HMG box (H131) and at a PTM site (K398), significantly upregulated expression of chondrogenic genes and potently induced chondrocytes from mouse embryonic fibroblasts (MEFs). The H131A/K398A variant remained unsumoylated in cells and exhibited a stronger DNA-binding activity than wild-type SOX9, especially when complexed with other proteins. Our results show that the novel SOX9 variant may be useful for efficient induction of chondrocytes and illuminate the strategic feasibility of mutating a transcription factor at functionally critical residues to expedite discovery of an optimized reprogramming factor.

Mucosal Application of a Low-Energy Electron Inactivated Respiratory Syncytial Virus Vaccine Shows Protective Efficacy in an Animal Model.

Respiratory syncytial virus (RSV) is a leading cause of acute lower respiratory tract infections in the elderly and in children, associated with pediatric hospitalizations. Recently, first vaccines have been approved for people over 60 years of age applied by intramuscular injection. However, a vaccination route via mucosal application holds great potential in the protection against respiratory pathogens like RSV. Mucosal vaccines induce local immune responses, resulting in a fast and efficient elimination of respiratory viruses after natural infection. Therefore, a low-energy electron irradiated RSV (LEEI-RSV) formulated with phosphatidylcholine-liposomes (PC-LEEI-RSV) was tested ex vivo in precision cut lung slices (PCLSs) for adverse effects. The immunogenicity and protective efficacy in vivo were analyzed in an RSV challenge model after intranasal vaccination using a homologous prime-boost immunization regimen. No side effects of PC-LEEI-RSV in PCLS and an efficient antibody induction in vivo could be observed. In contrast to unformulated LEEI-RSV, the mucosal vaccination of mice with PC formulated LEEI-RSV showed a statistically significant reduction in viral load after challenge. These results are a proof-of-principle for the use of LEEI-inactivated viruses formulated with liposomes to be administered intranasally to induce a mucosal immunity that could also be adapted for other respiratory viruses.

Structure-activity relationship of 2,4-D correlates auxinic activity with the induction of somatic embryogenesis in Arabidopsis thaliana.

2,4-dichlorophenoxyacetic acid (2,4-D) is a synthetic analogue of the plant hormone auxin that is commonly used in many in vitro plant regeneration systems, such as somatic embryogenesis (SE). Its effectiveness in inducing SE, compared to the natural auxin indole-3-acetic acid (IAA), has been attributed to the stress triggered by this compound rather than its auxinic activity. However, this hypothesis has never been thoroughly tested. Here we used a library of forty 2,4-D analogues to test the structure-activity relationship with respect to the capacity to induce SE and auxinic activity in Arabidopsis thaliana. Four analogues induced SE as effectively as 2,4-D and 13 analogues induced SE but were less effective. Based on root growth inhibition and auxin response reporter expression, the 2,4-D analogues were classified into different groups, ranging from very active to not active auxin analogues. A halogen at the 4-position of the aromatic ring was important for auxinic activity, whereas a halogen at the 3-position resulted in reduced activity. Moreover, a small substitution at the carboxylate chain was tolerated, as was extending the carboxylate chain with an even number of carbons. The auxinic activity of most 2,4-D analogues was consistent with their simulated TIR1-Aux/IAA coreceptor binding characteristics. A strong correlation was observed between SE induction efficiency and auxinic activity, which is in line with our observation that 2,4-D-induced SE and stress both require TIR1/AFB auxin co-receptor function. Our data indicate that the stress-related effects triggered by 2,4-D and considered important for SE induction are downstream of auxin signalling.

Agrobacterium rhizogenes-Mediated Genetic Transformation and Establishment of CRISPR/Cas9 Genome-Editing Technology in Limonium bicolor

Limonium bicolor is a perennial herbaceous plant belonging to the Plumbaginaceae family. It can be used as a dried flower or in cut flower arrangements and serves as a model recretohalophyte. Its genome sequencing has been recently completed. However, the research on L. bicolor is limited by the absence of a highly efficient genetic transformation system. In this study, we established a highly efficient Agrobacterium rhizogenes-mediated L. bicolor genetic transformation method. The transgenic hairy roots were induced from the hypocotyl of L. bicolor using A. rhizogenes strain K599 harboring pRdGa4Cas9 plasmid (which carries an expression cassette of 35S::DsRed2). The transgenic shoots were regenerated from hairy root segments (~0.1 cm diameter), and induction efficiency was achieved at 100%. The transgenic shoots with 4–5 rosette leaves were directly planted into the soil to induce the transgenic roots. Therefore, transgenic plantlets were produced. The DsRed2 can be used as a reliable reporter gene in screening transgenic plantlets. Furthermore, we also established a CRISPR/Cas9 system in L. bicolor employing the A. rhizogenes-mediated genetic transformation approach. The highly efficient transformation method and CRIPSP/Cas9 system established will provide a valuable tool for functional genomics investigation and trait improvement in L. bicolor.

The Optimization of Medium Conditions and Auxins in the Induction of Adventitious Roots of Pokeweed (Phytolacca americana L.) and Their Phytochemical Constituents.

Pokeweed, Phytolacca americana L., is considered a widely spreading invasive plant, while saponin contents accumulated in the roots have pharmaceutical uses, such as rheumatism treatments and anti-inflammation. Adventitious root cultures are an important source of diverse secondary metabolites, which have significant applications in various fields. This study focused on the optimization of parameters for root induction using different medium states and treatments with auxins on a pokeweed leaf. Semisolid and liquid MS (Murashige and Skoog, 1962) media were supplemented with indole-3-butyric acid (IBA) and 1-naphthylacetic acid (NAA) at 0.5, 1, 2, and 4 mg/L. Root growth parameters, e.g., induction percentage, root numbers, length, and weight, were measured to determine the adventitious root induction efficiency. Total phenolic content, total flavonoid content, total saponin content, and antioxidant activity were recorded. Results showed that adventitious roots induced in semisolid MS medium supplemented with 0.5 mg/L NAA exhibited a high density of lateral roots. Appropriate medium state and auxin for adventitious root induction in pokeweed were determined as semisolid medium supplemented with 2 mg/L NAA. Considering phytochemicals, adventitious roots induced in liquid medium containing 0.5-1 mg/L NAA had the highest yield extract percentage. Additionally, adventitious roots cultivated in a liquid medium enriched with 1 mg/L NAA exhibited the highest phenolic and saponin contents. A principal component analysis (PCA) biplot and hierarchical cluster analysis (HCA) heatmap demonstrated different response patterns between semisolid and liquid media applied with NAA. The results of the semisolid media were grouped together due to high expression levels of the root induction parameters, while elevated phytochemical values were observed in the liquid media treatments. The results suggested two different media that provide the highest adventitious root induction efficiency and the greatest phytochemical contents: semisolid medium with 2 mg/L NAA and liquid medium with 1 mg/L NAA, respectively. These culture media can be applied to optimize adventitious root culture of pokeweed and in vitro phytochemical production.

Callus Derived from Petals of the Rosa hybrida Breeding Line 15R-12-2 as New Material Useful for Fragrance Production.

Rose (Rosa hybrida) is a major flower crop worldwide and has long been loved for its variety of colors and scents. Roses are mainly used for gardening or cutting flowers and are also used as raw materials for perfumes, cosmetics, and food. Essential oils, which are extracted from the flowers of plants, including roses, have various scents, and the essential oil market has been growing steadily owing to the growing awareness of the benefits of natural and organic products. Therefore, it is necessary to develop a system that stably supplies raw materials with uniform ingredients in line with the continuous increase in demand. In this study, conditions for the efficient induction of callus were established from the petals of the rose breeding line 15R-12-2, which has a strong scent developed by the National Institute of Horticultural and Herbal Science, Rural Development Administration. The highest callus induction rate (65%) was observed when the petals of the fully open flower (FOF) were placed on the SH11DP medium so that the abaxial surface was in contact with the medium. In addition, the VOCs contained in the petals of 15R-12-2 and the petal-derived callus were analyzed by HS-SPME-GC-MS. Thirty components, including esters and alcohols, were detected in the petal-derived callus. Among them, 2-ethylhexan-1-ol, which showed 59.01% relative content when extracted with hexane as a solvent, was the same component as detected in petals. Therefore, petal-derived callus is expected to be of high industrial value and can be suggested as an alternative pathway to obtaining VOCs.

TRPML3/BK complex promotes autophagy and bacterial clearance by providing a positive feedback regulation of mTOR via PI3P

TRPML3 is a Ca2+/Na+ release channel residing in both phagophores and endolysosomal membranes. It is activated by PI3P and PI3,5P2. Its activity can be enhanced by high luminal pH and by replacing luminal Na+ with K+. Here, we report that big-conductance Ca2+-activated potassium (BK) channels form a positive feedback loop with TRPML3. Ca2+ release via TRPML3 activates BK, which in turn facilitates TRPML3-mediated Ca2+ release, potentially through removing luminal Na+ inhibition. We further show that TRPML3/BK and mammalian target of rapamycin (mTOR) form another positive feedback loop to facilitate autophagy induction in response to nutrient starvation, i.e., mTOR inhibition upon nutrient starvation activates TRPML3/BK, and this further reduces mTOR activity, thereby increasing autophagy induction. Mechanistically, the feedback regulation between TRPML3/BK and mTOR is mediated by PI3P, an endogenous TRPML3 activator that is enriched in phagophores and is up-regulated by mTOR reduction. Importantly, bacterial infection activates TRPML3 in a BK-dependent manner, and both TRPML3 and BK are required for mTOR suppression and autophagy induction responding to bacterial infection. Suppressing either TRPML3 or BK helps bacteria survival whereas increasing either TRPML3 or BK favors bacterial clearance. Considering that TRPML3/BK is inhibited by low luminal pH but activated by high luminal pH and PI3P in phagophores, we suggest that TRPML3/BK and mTOR form a positive feedback loop via PI3P to ensure efficient autophagy induction in response to nutrient deprivation and bacterial infection. Our study reveals a role of TRPML3-BK coupling in controlling cellular homeostasis and intracellular bacterial clearance via regulating mTOR signaling.

Effect of Plant Growth Regulators on Callus Induction of Black Rice (Oryza sativa L.)

Aims: Black rice of Manipur is known for its aroma and nutritional quality. As per the reports on tissue culture of rice, callus induction response of traditional cultivars was much lower than the modern high yielding varieties. Considering the importance of the black rice, and effort was taken to standardized callus induction protocol of black rice of Manipur.
 Study Design: Complete Randomized Design.
 Place and Duration of Study: Department of Seed Science and Technology, Faculty of Agriculture, Uttar Banga Krishi Viswavidyalaya, Pundibari, Cooch Behar 736165, West Bengal, India, between June 2020 and July 2023.
 Methodology: Mature seeds of two black rice varieties used in this work. The seeds were dehusked manually, washed with running tap water. Then sterilized with 1% bavistin and again washed with running tap water. Seeds were surface sterilized with 0.2% HgCl2 for 15 minutes. Sterilised seeds were rinsed with sterile distilled water inside laminar air flow hood. The seeds were blotting dry using autoclaved tissue paper and sterile seeds were cultured with the scutellum pointing up wards on callus induction medium fortified with different concentrations and combinations of plant growth regulators.
 Results: MS medium fortified with 2,4-D was found to be better than picloram and TDZ to induce callus and the 2,4-D @ 1.00 mg/L and 2,4-D 2.00 mg/L was found to be the best concentrations. Callus health was also good when MS medium invigorated with 2,4-D.
 Conclusion: An efficient embryogenic callus induction and plantlet regeneration protocol for black rice was established by using mature seeds. In callus induction studies, with different concentrations of PGR, 2,4-D, higher callus induction frequency was observed with 1.00 mg/L 2,4-D and 2,4-D @ 2 mg/L for both rice varieties.

Delivery of chemotherapeutic drug targeting folate receptor to oral cancer cells using functionalized carbon nanospheres

Folate receptor (FR) (α) has long been the subject of active interest as regards its potential to serve as a target for cancer therapy. FR has been found to be overexpressed in several cancers, including clinical samples of different stages from OSCC (oral squamous cell carcinoma) patients. However, no clear correlation or conclusive finding has been obtained so far which might indicate the efficacy of FR as a credible target for the treatment of OSCC. All cell lines to be used were assessed for FR-expression. Subsequently, we developed glucose-derived carbon nanospheres (CSPs) and primed them with a Folate-based cationic lipid FA8 and the chemotherapeutic drug doxorubicin (DOX). CSP based delivery systems along with pristine drug DOX were characterized and treated subsequently to in vitro cultures of OSCC cells and assessed for cancer cell targetability as well as cell death. Subsequently, treatment was administered to immunocompetent C57 mice carrying MOC2 based syngeneic OSCC tumours and assessed for tumour regression and toxicity. Ligand primed targeted CSPs exhibited commendable drug uptake as well as efficient induction of cell death. Further, receptor blocking studies revealed FR-mediated uptake, preferentially in cancer cells. Drug once delivered by ligand-primed CSPs was retained longer inside cells than pristine drug alone, indicating possibilities of better therapeutic outcome. In animal studies, CSP-FA8-DOX (Ligand primed targeted CSP) demonstrated significant regression in tumour size compared to pristine DOX as well as CSP-DOX (non-targeted CSP) treated animals. FR-mediated system CFD demonstrated targeted drug uptake and apoptotic death selectively in cancer cells. Significant tumour regression was also observed in vivo. Overall, it may be presumed that the FR is a therapeutic target with substantial potential in OSCC treatment.

The efficient induction of human retinal ganglion-like cells provides a platform for studying optic neuropathies

Retinal ganglion cells (RGCs) are essential for vision perception. In glaucoma and other optic neuropathies, RGCs and their optic axons undergo degenerative change and cell death; this can result in irreversible vision loss. Here we developed a rapid protocol for directly inducing RGC differentiation from human induced pluripotent stem cells (hiPSCs) by the overexpression of ATOH7, BRN3B, and SOX4. The hiPSC-derived RGC-like cells (iRGCs) show robust expression of various RGC-specific markers by whole transcriptome profiling. A functional assessment was also carried out and this demonstrated that these iRGCs display stimulus-induced neuronal activity, as well as spontaneous neuronal activity. Ethambutol (EMB), an effective first-line anti-tuberculosis agent, is known to cause serious visual impairment and irreversible vision loss due to the RGC degeneration in a significant number of treated patients. Using our iRGCs, EMB was found to induce significant dose-dependent and time-dependent increases in cell death and neurite degeneration. Western blot analysis revealed that the expression levels of p62 and LC3-II were upregulated, and further investigations revealed that EMB caused a blockade of lysosome–autophagosome fusion; this indicates that impairment of autophagic flux is one of the adverse effects of that EMB has on iRGCs. In addition, EMB was found to elevate intracellular reactive oxygen species (ROS) levels increasing apoptotic cell death. This could be partially rescued by the co-treatment with the ROS scavenger NAC. Taken together, our findings suggest that this iRGC model, which achieves both high yield and high purity, is suitable for investigating optic neuropathies, as well as being useful when searching for potential drugs for therapeutic treatment and/or disease prevention.

On the efficient implementation of classification rule learning

AbstractRule learning methods have a long history of active research in the machine learning community. They are not only a common choice in applications that demand human-interpretable classification models but have also been shown to achieve state-of-the-art performance when used in ensemble methods. Unfortunately, only little information can be found in the literature about the various implementation details that are crucial for the efficient induction of rule-based models. This work provides a detailed discussion of algorithmic concepts and approximations that enable applying rule learning techniques to large amounts of data. To demonstrate the advantages and limitations of these individual concepts in a series of experiments, we rely on BOOMER—a flexible and publicly available implementation for the efficient induction of gradient boosted single- or multi-label classification rules.

An improved in-vitro regeneration protocol using scutellum of mature and immature embryos of wheat

Generally, conventional practices take a long time to improve agronomic traits in common wheat (Triticum aestivum L.). However, biotechnological tools provide the best opportunity to enhance crop potential within a limited time. A simple and repeatable protocol for wheat regeneration is the priority for transforming the genetic makeup of elite wheat cultivars. Among 10 types of callus induction media (CIM), CIM4 with supplemented Murashige and Skoog (MS) media+ dicamba (2 mg/l) has 98.22 and 97.33% callus induction efficiency (CIE) in immature and mature scutellum, respectively. Precocious germination, considered a key barrier during callus induction, was fully controlled by excising the embryonic axis. Among 29 different shoot induction media (SIM) assessed in the study, SIM 29 containing zeatin 5 mg/l, 6-benzylaminopurine (BAP) 5 mg/l, 2,4-dichlorophenoxyacetic acid (2,4-D) 0.25 mg/l,1-naphthaleneacetic acid (NAA) 0.25 mg/l, and copper sulfate (CuSO4) 20 mg/l was found most responsive for shoot induction. The combined effect of cytokinin and CuSO4 resulted in an improved wheat regeneration response in two popular Indian wheat varieties, namely HD2967 and HD3171. Rooting was observed at 100% using IAA and IBA @ 0.2 mg/l concentration in root induction media (RIM). The whole in-vitro regeneration protocol from embryo to transfer in the greenhouse took 8 to 9 weeks. The in-vitro regeneration protocol developed in the present study could further be utilized to improve the genetic makeup of elite wheat cultivars utilizing genetic engineering tools.

Enhanced production of bioactive plumbagin in hairy root cultures and adventitious root cultures of Plumbago zeylanica L. by a novel apocarotenoid elicitor, α-ionone

Plumbagin, a medicinally important secondary metabolite is produced in the roots of Plumbago zeylanica Linn. Natural harvest of plumbagin from roots is a destructive process that destroys the entire plant. The aim of this study was the in vitro production of plumbagin at high concentrations. For this purpose, hairy root cultures (HRCs) and adventitious root cultures (ARCs) were induced from P. zeylanica leaf explants. Additionally, first-time α-ionone was used as an elicitor to trigger plumbagin biosynthesis. HRCs were developed using the two different strains of Agrobacterium rhizogenes (LBA1334, and R1000). R1000 strain showed the highest HR induction efficiency. ARCs were induced from the leaf explants growing in Murashige and Skoog (MS) solid media supplemented with 1 mg/L indole-3-butyric acid. Newly emerged adventitious roots were used to develop root suspension cultures in a liquid MS medium. Plumbagin content was determined by HPLC/DAD. Both HRCs and ARCs showed fast growth kinetics. The greatest increase in the production of plumbagin was achieved with 10 µM α-ionone. As compared to untreated controls, the optimum α-ionone dose resulted in a 3.6-fold and 3.1-fold increase in plumbagin in HRCs, and ARCs, respectively. These results support that α-ionone elicitor-treated HRCs and ARCs are promising technology for improved production of plumbagin to meet commercial demand.

Inducing Pseudopregnancy in Female Mice Without the Need for Vasectomized Males Prior to Non-Surgical Embryo Transfer or Artificial Insemination.

For successfully maintaining pregnancy with embryo transfer or artificial insemination,female recipient mice must be induced into a pseudopregnant state. Female mice are traditionally paired overnight with vasectomized males, and the following morning, the presence of a copulation plug is assessed. To increase the efficiency of producing pseudopregnant females, a cervical manipulation technique has been standardized to be used in combination with non-surgical embryo transfer or artificial insemination techniques in mice. The blunt end of a small plastic rod is inserted vaginally to contact the cervix and is vibrated for 30 s by contact with a trimmer. The procedure is quick and does not require anesthesia or analgesia. This technique increases the reliability and predictability of producing pseudopregnant females and entirely eliminates the requirement for vasectomized males. For CD1 mice, the efficiency of pseudopregnancy induction using cervical manipulation was 83% for females in estrus (N = 76) but only 38% of females in estrus were plugged by vasectomized males (N = 24). Artificial insemination in CD1 mice was performed by estrus synchronization with hormones, cervical manipulation, and the uterine transfer of sperm. Artificial insemination recipients receiving cervical manipulation (N = 76) had a pregnancy rate of 72% and an average litter size of 8.3 pups. This method can also be used to produce pseudopregnant females for non-surgical embryo transfer. Therefore, inducing pseudopregnancy by cervical manipulation is a convenient and efficient alternative to mating with a vasectomized male when performing non-surgical assisted reproduction techniques. Using cervical manipulation provides 3Rs (replacement, reduction, and refinement) benefits for assisted reproduction techniques by reducing the number of animals required and eliminating the necessity for surgically altered males.

The hairy root induction efficiency and that plumbagin of Plumbago auriculata L.

The hairy root induction efficiency and that plumbagin of Plumbago auriculata L.