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The hawthorn yam (Dioscorea rotundata L.) is essential for food security and the economy in developing countries, but its cultivation faces critical challenges due to biotic, abiotic, and physiological limitations that affect its commercial quality. Salicylic Acid (SA) is proposed as a plant growth regulator due to its positive effects on morpho-physiological processes. This study evaluates the morphogenetic response of SA and Acetylsalicylic Acid (ASA) in the in vitro culture of yam to improve its quality in terms of development and biomass. Murashige and Skoog (MS) media with different concentrations of SA and ASA (0, 5, 10, 20, and 30 μM) and a pH adjusted to 5.8 were used. The explants were cultured under controlled conditions and evaluated at specific time intervals (8, 16, 30, 60, and 75 days), recording the number of nodes, shoots, leaves, roots, and plant height. Variance analysis (ANOVA) and Tukey's test (p≤ 0.05) were applied using R version 4.3.0. The results showed that 5 μM of ASA and 30 μM of SA significantly improved plant height, the number of leaves, and roots. These findings suggest that SA and ASA can increase biomass and the commercial quality of yam grown in vitro, indicating a potential strategy to sustainably improve agricultural production. [ABSTRACT FROM AUTHOR]

Manihot esculenta species is a crucial tuber crop and the fourth main source of calories in the tropics, providing 250 kcal/ha/day and being a staple food for more than 800 million people worldwide. This study consists of improving the growth and development of cassava in vitro by means of the effect of salicylic acid (SA) and its commercial derivative, acetylsalicylic acid (ASA or "Aspirin"), on two varieties of cassava that are popular in the region: "Verdecita" and "Chirosa negrita". Ten treatments were established for the two varieties, with 12 replicates each one, using culture media with MS salts, sucrose, myo-inositol, thiamine and gelrite. Treatments included concentrations of SA and ASA (0, 10, 20, 30 and 40 µM), adjusting pH to 5.8 prior to sterilization. The 1 cm nodal segments were incubated at 28 ± 5 °C, 65% relative humidity and a 12-hour photoperiod. At 60 days, treatment with 20 µM SA resulted in increased stem growth, number of roots and leaves, while higher concentrations caused phytotoxicity. All SA treatments (>30 µM) resulted in phytotoxicity and explant death, with minimal growth at concentrations below 30 µM. In conclusion, concentrations lower than 30 µM of SA and ASA enhance morphogenetic responses in Manihot esculenta in vitro and higher concentrations affect explant survival. It is thus observed that SA, in general, stimulates better biomass production when compared to ASA. [ABSTRACT FROM AUTHOR]

Persistent mating-induced endometritis is a major cause of poor fertility rates in the mare. Endometritis can be investigated using an ex vivo equine endometrial explant system which measures uterine inflammation using prostaglandin F2α as a biomarker. However, this model has yet to undergo a wide-ranging assessment through transcriptomics. In this study, we assessed the transcriptomes of cultured endometrial explants and the optimal temporal window for their use. Endometrium harvested immediately post-mortem from native pony mares (n = 8) were sampled (0 h) and tissue explants were cultured for 24, 48 and 72 h. Tissues were stored in RNALater, total RNA was extracted and sequenced. Differentially expressed genes (DEGs) were defined using DESeq2 (R/Bioconductor). Principal component analysis indicated that the greatest changes in expression occurred in the first 24 h of culture when compared to autologous biopsies at 0 h. Fewer DEGs were seen between 24 and 48 h of culture suggesting the system was more stable than during the first 24 h. No genes were differentially expressed between 48 and 72 h but the low number of background gene expression suggested that explant viability was compromised after 48 h. ESR1, MMP9, PTGS2, PMAIP1, TNF, GADD45B and SELE genes were used as biomarkers of endometrial function, cell death and inflammation across tissue culture timepoints. STRING assessments of gene ontology suggested that DEGs between 24 and 48 h were linked to inflammation, immune system, cellular processes, environmental information processing and signal transduction, with an upregulation of most biomarker genes at 24 h. Taken together our observations indicated that 24–48 h is the optimal temporal window when the explant model can be used, as explants restore microcirculation, perform wound healing and tackle inflammation during this period. This key observation will facilitate the appropriate use of this as a model for further research into the equine endometrium and potentially the progression of mating-induced endometritis to persistent inflammation between 24 and 48 h.

Taraxacum officinale (L.) Weber ex F.H. Wigg, commonly known as dandelion, is a cosmopolitan and perennial weed, which has medicinal properties. In vitro propagation methods are widely used on plants that have difficulties in cultivation and, consequently, low extraction yields of active metabolites. Thus, callus culture has been considered to be useful for the accumulation of several metabolites. In this study, we aimed to establish an efficient protocol for callus induction and maintenance of T. officinale, for which explant type, carbon source, light conditions, and nine different combinations of plant growth regulators (PGRs), such as 1-naphthaleneacetic acid (NAA) (from 0.05 to 0.5 mg/L) and 6-benzylaminopurine acid (BAP) (from 0.5 to 3.0 mg/L), were evaluated. The results showed that hypocotyls and roots from sterile seedlings are the best sources for callus induction, with 100% of callogenesis at every condition tested, and more than 95% of viability and friability. Complete darkness and a medium supplemented with sucrose at 2.3% (w/v) and 0.5 mg/L of NAA and 0.5 mg/L of BAP were the best conditions for callus induction, showing callus with low organogenesis and high friability. This study provides a basis for future studies on improving large-scale callus propagation and further establishment of suspension culture systems for commercial purposes.

Effective breeding of the main forest-forming gymnosperm species of Ukraine (Pinus sylvestris L., Larix decidua Mill., Abies alba Mill. And Risea abies (L.) Karsten), which are of significant value for Forestry and Landscape Architecture, is a very important task. The solution to this task may be the use of modern methods of reproduction, namely in vitro micro-cloning. Decontamination of explants was conducted applying the following sterilizing agents in different concentrations and in different combinations: tapwater (H2O) withadetergent; Medicalethanol (C2H5OH); sodiumhypochlorite (NaClO); silvernitrate (AgNO3). To prevent internal infections, a 0.1% "Imaninum" antibiotic was applied. To initiate explants, the following nutrient media were used: Murashige and Skoog (MS), Risser and White (RW) and Litvay (LM), which were modified with phytohormones: 2,4-D; NАА; IВА and BAP. Rooting of the obtained clones was performed on 1/2 MS and 1/2 LM media, supplemented with auxin (2,4-D; NАА IАА and the IВА). Adaptation of clones to ex vitro conditions was exercised in the sod soil and sand (1: 1) and sod soil with peat and sand (1: 1: 1).Thefollowingsterilizationresultswereobservedin: PinussylvestrisL. – 98, Larixdecidua Mill. – 96 Abiesalba Mill. – 98 and Riseaabies (L.) Karsten– 97 % ofasepticexplants. The proper selection of nutrient medium resulted in high initiation of: Pinus sylvestris L. – 92, Larix decidua Mill. – 95 Abies alba Mill. – 94 and Risea abies (L.) Karsten – 95%. The importance of the auxin and cytokinin presence in the medium at this stage should be noted. Rhizogenesis results of the study species are as follows: Pinus sylvestris L. – 86, Larix decidua Mill. – 91 Abies alba Mill. – 93 and Risea abies (L.) Karsten – 89%. It is established that injection of IАA to the medium is necessary, since the number of rooted clones was only increasing during all scenarios. The largest number of adapted clones was observed on the substrate of sod soil with peat and sand, except for the plant-regenerants of Larix decidua, where the substrate of the sod soil and sand had given a better result. Application of in vitro breeding for the study species is very promising, since satisfactory results were obtained at all stages. A further study of cultivation phase is necessary, since it will enable to determine the rate and stage of adaptation to in vivo conditions.

This study was conducted to examine explant density and source on production of in vitro potato microtubers, as well as survey of different newly-introduced potato cultivars for their microtuberization capacity at the Plant Tissue Culture Facility of the Department of Horticulture, Suez Canal University between 2014 until 2016. Results indicated that as culture density from single node explants increased, microtuber number and yield/petri dish were also increased. However, at the lowest density (10 explants/petri dish), the % tuberization and the average microtuber weight increased significantly over the higher densities (20, 25 and 30 explants/petri dish). Potato microtuber production from plantlets grown in vitroon microtuberization induction medium (liquid over solid media) was also studied using 5, 10 and 15 plantlets per jar.Results showed that microtuber number and yield/jar increased gradually with increasing plantlet density. However, on per plantlet basis, microtuber number increased at the lowest density. Microtuber yield/jar increased significantly at the highest density (1099 mg/jar) compared to 563.6 mg/jar at the lowest density. Average microtuber weight was also affected by culture density and genotype. At the lowest density, microtubers were significantly heavier than at the high density, and the cv. 'Safrane' recorded the highest weight per microtuber. Regarding the effect of explant source on microtuber production, results indicated that the use of single node explants derived from the top of the plantlet produced more microtubers than those taken from the basal part, or 3 node segments. Explant source also affected microtuber yield/dish. Single nodes from the top produced the highest yield compared to 3-nodes segment from the top (205 vs. 104.8 mg). The highest mean microtuber fresh weight was obtained from the culture of 3-node layered segment from the plantlet base, while single nodes from the top recorded lower microtuber fresh weight. The highest microtuber fresh weight was recorded in cv 'Universa' (114.3 mg) using single nodes from the plantlet base. Differences among the tested potato cultivars in microtuber production were detected.

Date palm ( Phoenix dactylifera L.) is the source of fruit and palm oil, which is broadly used in the food industry. The regeneration of recalcitrant date palm genotypes through somatic embryogenesis or organogenesis is difficult to achieve. Micropropagation, however, provides a way to obtain a sufficient number of true-to-type elite, healthy (disease-free) and vigorous plants to satisfy local and international markets. This mini-review highlights some of the main achievements in the tissue culture of date palm. In particular, explant selection and disinfection, shoot induction, multiplication and elongation, root induction and acclimatization are highlighted. In addition to using the in vitro tissue culture as the basis for an in vitro gene banking, a mid-term low-temperature storage of germplasm is possible by careful selection of the osmotic agent. A long-term storage of date palm using cryopreservation, with or without synthetic seeds is also possible. Molecular markers, as well as sex-specific markers, have been useful to discriminate germplasms and to identify somaclonal variants derived from tissue cultures. The genetic transformation of date palm can be achieved by either particle bombardment or Agrobacterium –based protocols. Thus, biotechnology is an important element of date palm germplasm development and its sustainable improvement.

Gerbera (Gerbera jamesonii Bolus ex Hook f.) is an important ornamental plant commodity with high economical value in Indonesia; however, development of the plant in larger scale commercially is constrained by the availability of qualified-planting materials. Conventional propagation is clearly not suitable to overcome the problem and therefore in vitro propagation protocol is importantly addressed. In vitro propagation protocol of G. jamesonii was successfully established in the study. Different explants (shoot tips, young leaves, petioles and petals) were selected and cultured on Murashige and Skoog (MS) basalt medium containing different plant growth regulators. Shoot tips and half-strength MS medium containing 1.5 mg l2017-06-20-12017-06-20 thidiazuron (TDZ) and 0.25 mg l2017-06-20-12017-06-20 N6-benzylaminopurine (BAP) were suitable explant and initiation medium for shoot formation with 75% explant regeneration and 5.5 shoots produced per explant. Shoots derived from selection stage were proliferated on half-strength MS medium supplemented with 0.25 mg l2017-06-20-12017-06-20 BAP and resulting higher shoot regeneration up to 7.5 shoots produced per shoot with 22.8 leaves and 0.83 cm leaf length. Multiplication of shoots on the half-strength MS medium fortified by 0.25 mg l2017-06-20-12017-06-20 BAP increased gradually from the first subculture till the fifth subculture with 9.1 shoots produced per shoot subcultured and reduced thereafter. Shoots were then rooted on half-strength MS medium augmented with 0.5 mg l2017-06-20-12017-06-20 α-naphthalene acetic acid (NAA) and 1.5 g l2017-06-20-12017-06-20 activated charcoal (AC) and produced 2.1 roots per shoot with 2.52 cm length of roots. Well-rooted shoots were acclimatized in a mixture of burned-rice husk and organic manure (1:1, v/v) with 95% survival rate and 114 plantlets grew well under this treatment.

Composite delivery systems where drugs are electrospun in different layers and vary the drug stacking-order are posited to affect bioavailability. We evaluated how the formulation characteristics of both burst- and sustained-release electrospun fibers containing three physicochemically diverse drugs: dapivirine (DPV), maraviroc (MVC) and tenofovir (TFV) affect in vitro and ex vivo release. We developed a poly(hydroxyethyl methacrylate) (pHEMA) hydrogel release platform for the rapid, inexpensive in vitro evaluation of burst- and sustained-release topical or dermal drug delivery systems with varying microarchitecture. We investigated properties of the hydrogel that could recapitulate ex vivo release into nonhuman primate vaginal tissue. Using a dimethyl sulfoxide extraction protocol and high-performance liquid chromatography analysis, we achieved >93% recovery from the hydrogels and >88% recovery from tissue explants for all three drugs. We found that DPV loading, but not stacking order (layers of fiber containing a single drug) or microarchitecture (layers with isolated drug compared to all drugs in the same layer) impacted the burst release in vitro and ex vivo. Our burst-release formulations showed a correlation for DPV accumulation between the hydrogel and tissue (R2= 0.80), but the correlation was not significant for MVC or TFV. For the sustained-release formulations, the PLGA/PCL content did not affect TFV release in vitro or ex vivo. Incorporation of cells into the hydrogel matrix improved the correlation between hydrogel and tissue explant release for TFV. We expect that this hydrogel-tissue mimic may be a promising preclinical model to evaluate topical or transdermal drug delivery systems with complex microarchitectures.

Slaughter products are good source of bioactive substances with different biological effects. Nowadays, approximately 220 functional peptides consisting of 2–10 amino acid residues have been identified. In this study, the biological activity of medium-molecular-weight substances (5–30 kDa, Mw) and low-molecular-weight substances (less than 5 kDa, Mw) extracted from aorta of Sus scrofa were investigated in vitro. Medium-molecular-weight and low-molecular-weight fractions possessed a tissue specific effect on organotypic tissue cultures of chicken embryos and laboratory rats. Biological activity depended on molecular weight. Medium-molecular-weight fractions possessed higher effect on aortas explants of chicken embryo: area index of cellular monolayer was 56.6±5.2 % (area index of the monolayer was 37.47±3.27 % for low-molecularweight fraction) in comparison with control explants. The low-molecular-weight fractions stimulated aortic tissues explants of aging rats — area index exceeded area index formedium-molecularweight fractions by more than 30%.

Background Cellular therapy clinical applications require large-scale production of stem cells. Therefore, abundance, ease of isolation, and proliferative potential are the most important factors in choosing the appropriate source of cells for transplantation studies. Multipotent stem cells obtained from periodontal ligament (PDL) can be used in periodontal tissue regeneration. In this study, we aimed to evaluate and compare the characteristics of periodontal ligament stem cells (PDLSCs), extracted by either enzymatic digestion or explant methods, and expanded using two different serum types: fetal bovine serum (FBS) and xeno-free platelet lysate (PL). Methods Expanded PDLSCs were assessed for their proliferation capacity, surface markers expression, colony formation, differentiation potential and ability to self-renewal. Most importantly, PDLSCs were evaluated for their ability to produce osteoblasts in vitro. Results PDLSCs isolated by explant method and expanded in PL serve as a promising source of stem cells for osteoblasts regeneration. These cells showed higher proliferation capacity, they retained their stemness characteristics throughout the passages and they revealed an increase in the expression level of osteogenic markers, without showing any karyotypic abnormalities after cell expansion. Conclusions PDLSCs produced using explant extraction method and expanded in cell culture media supplemented with PL provide an excellent source of xeno-free cells for the generation of functional osteoblasts.

Abstract:A simple procedure for direct embryogenesis without an intervening callus production was developed in valerian (Valeriana officinalis L.) using the leaf segments. Direct somatic embryogenesis were induced using by half-strength MS medium supplemented with 2,4-D (0.5 mg·l-1), glutamic acid (100 mg·l-1), 4% sucrose and 8 g·l-1 agar. Germinated embryo to form plantlets enhanced on MS medium supplemented with NAA (0.1 mg·l-1) and Kin (2 mg·l-1). Regenerated plants with well-developed root and shoot systems were successfully (72%) transferred to greenhouse.

Models have been extensively used to investigate disease pathogenesis. Animal models are costly and require extensive logistics for animal care, and samples are not always suitable for different analytical techniques or to answer the research question. In vitro cell culture models are generally focused on recreating a specific characteristic of an organ and are limited to a single cell population that does not display the characteristic tissue architecture of the source organ. In addition, such models do not account for the many interactions between pathogens and the diverse cell subsets that are normally present in a given organ. Conclusions based on conventional 2D cell culture methods are limited, requiring extrapolation from a reductionist model to understand in vivo events. In vitro organ culture (IVOC) offers a way to overcome some of these limitations. Explants conserve important in vivo characteristics, such as different cell types and complex tissue architecture. This in vitro (ex vivo) organ culture protocol of the swine large intestine aims at maintaining viable colonic mucosa for up to 5 days. The protocol described herein applies a combination of methods used for immortalized cell culture and stem cell stimulation to support the physiological cellular flow inherent of the intestinal mucosa. Required equipment includes a hyperoxic chamber and culture at the air-liquid interface., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Outcome optimization after the placement of a spinal cord stimulator (SCS) is critical. The objective of this study was to determine if an association existed between pre-procedural opioid use (compared to patients who were opioid-naïve) and postoperative long-term outcomes following SCS placement., Objective: To examine the impact of preprocedural opioid use on long-term outcomes after SCS therapy., Study Design: Cohort study utilizing a nationwide database., Setting: Retrospective., Methods: With the use of data from HCA Healthcare's national database, a retrospective cohort study was performed to analyze differences in outcomes between opioid-naïve patients and preoperative opioid users who underwent SCS placements. The primary outcome of interest was device explantation at 6 months and 12 months. Secondary outcome measurements included reoperations and readmissions at 6 months and 12 months, as well as operative complications. Multivariable logistic regression models were performed to analyze the association of preoperative opioid use with those outcomes. The odds ratio (OR), 95% confidence intervals (CI), and P values were reported for the independent variables., Results: The final study population consisted of 13,893 patients who underwent SCS placements. In univariate analyses, patients who used opioids preoperatively had higher 6-month (3.6% vs. 2.6%) and one-year removal rates (3.6% vs. 2.8%) (all P < 0.009). On multivariable logistic regression, those using opioids preoperatively had higher odds of removal at 6 months (OR = 1.290, 95% CI 1.05-1.58, P = 0.01) and at one year (OR = 1.23, 95% CI 1.01-1.50, P = 0.04). There was no difference between patients requiring preoperative opioids and patients who were opioid-naive as far as the odds of 6- or 12-month readmissions were concerned. Compared to the opioid-naive group, patients requiring preoperative opioids had increased odds of reoperation at 6 months (OR = 1.2, 95% CI 1.02-1.40, P = 0.03). There were no differences in the odds of complications between both cohorts., Limitations: Opioid use in this study was defined as using opioids preoperatively in the 30 days leading up to surgery., Conclusion: Patients requiring preoperative opioids before SCS placements had increased odds of SCS explantation at 6 months and 12 months, as well as increased odds of reoperation at 6 months.

Adipose tissue provides a valuable cell source for tissue engineering, regenerative medicine, and adipose tissue biology studies. The most widely used adipose-derived stromal/stem cells (ASCs) isolation protocol involves enzymatic digestion with collagenase. However, the yield of the method often proves to be poor if not impossible for collection of sufficient stromal vascular fraction (SVF) for expansion when the sample size is small, for instance when only newborn mice are available for cell culture. Here, we describe an efficient protocol for the isolation and expansion of ASCs using explant culture as an alternative. Briefly, adipose tissue was minced after removing excess liquid. Then, the minced tissue was placed in culture dishes or flasks. The cells will migrate out of tissue and adhere to the culture surface after one or more days., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Zika virus (ZIKV) is an emerging pathogen that can cause microcephaly and other neurological defects in developing fetuses. The cellular response to ZIKV in the fetal brain is not well understood. Here, we show that ZIKV infection of human fetal astrocytes (HFAs), the most abundant cell type in the brain, results in elevated expression and secretion of fibroblast growth factor 2 (FGF2). This cytokine was shown to enhance replication and spread of ZIKV in HFAs and human fetal brain explants. The proviral effect of FGF2 is likely mediated in part by suppression of the interferon response, which would represent a novel mechanism by which viruses antagonize host antiviral defenses. We posit that FGF2-enhanced virus replication in the fetal brain contributes to the neurodevelopmental disorders associated with in utero ZIKV infection. As such, targeting FGF2-dependent signaling should be explored further as a strategy to limit replication of ZIKV. [ABSTRACT FROM AUTHOR]

Cul􀆟va􀆟on of rare flora species in order to preserve their gene pool is one of the main tasks of our 􀆟me. In vitro methods, in combina􀆟on with ex situ, are nowadays becoming increasingly important means of suppor􀆟ng and maintaining the level of phytodiversity stability. Eucommia ulmoides Oliver is a deciduous tree, belongs to the Eucommiaccac family. This tree species grows in the territory of the NULES of Ukraine Arboretum in a single copy, is a valuable decora􀆟ve introducer and has a number of medicinal proper􀆟es. It is currently in poor condi􀆟on and needs treatment and reproduc􀆟on for preserva􀆟on. The purpose of the study was to develop biotechnology of microclonal propaga􀆟on of E. ulmoides Oliv plants in the ini􀆟al stages of reproduc􀆟on for using healthy plant-materials in landscaping and cul􀆟va􀆟on in botanical gardens. The relevance of microclonal reproduc􀆟on of E. ulmoides Oliv is substan􀆟ated. Features of introduc 􀆟on into culture of E. ulmoides Oliv were established with using various sterilizing agents, types of explants and growing condi􀆟ons. Different types of shoots 15-25 cm long with apical and lateral buds, selected from the donor plant in February - March were used for the work. Murasighe & Skoog (MS) and Driver (DKW) nutrient media were used for cul􀆟va􀆟on of Eucomia, with cytokinins and auxins addi􀆟on to their composi􀆟on: 6-BA (6-benzylaminopurine), NAC (α-naphthylace􀆟c acid) and IAA (β-indolyl-3-ace􀆟c acid) both individually and in combina􀆟on. Sucrose (0.3 %), mesoinositol (100 mg / l) and agar (0.7 %) were added to the medium, the pH of the medium was 5.7-5.8. The influence of different steriliza􀆟on variants on the development of Eucommia ulmoides Oliver microshoots has been studied. As a result of the research the technique of steriliza􀆟on of E. ulmoides Oliv explants with 78 % efficiency due to surface steriliza􀆟on of 0.1 % HgCl2 on annual shoots has been worked out. Op􀆟mal nutrient media were established at the stage of in vitro culture of E. ulmoides Oliv. In par􀆟cular, for the microclonal reproduc􀆟on of E. ulmoides Oliv., the nutrient medium MS 1 supplemented with 0.5 mg·l-1 BA and 0.1 mg·l-1 NAA is recommended in the ini􀆟al stages. [ABSTRACT FROM AUTHOR]

Background: Failure of endovascular aneurysm repair may require explant of the stent graft in a subset of patients. We sought to assess outcomes in a cohort of patients undergoing explant of endovascular aneurysm repair in both emergency and elective settings. Methods: Patients undergoing explant of endovascular aneurysm repair were identified from a prospectively maintained database, with additional information obtained through retrospective analysis of medical records. Results: Over a 21-year period, 1997–2018 (May), there were 597 endovascular aneurysm repair procedures performed in our institution for abdominal aortic aneurysm. There were 19 endovascular aneurysm repair explants; five of these were referrals from other vascular centres. The median age was 73 years (range 46–81). The median length of time from insertion to explant was 39.2 months (range 0–153). Indications for elective explant were type Ia endoleak (n = 4), type 1b endoleak (n = 1), type II endoleak with increasing sac size (n = 1), type I/III endoleak (n = 1), type IV endoleak (n = 1), and increasing sac size without evident endoleak (type V, n = 2). The remaining nine cases were emergency procedures, with four patients presenting with rupture post endovascular aneurysm repair, four patients presenting with acute stent thrombosis, of which one also had a type 1a endoleak and one aorto-enteric fistula. There were no mortalities in the elective group and three mortalities in the emergency group (0 vs 33.3%, p = 0.087). Overall 30-day mortality was 15.8% Conclusion: Explant of aortic stent grafts can be associated with high mortality and morbidity rates, especially in the emergent setting. Patient and device selection and post-operative surveillance remain vitally important to optimise outcomes post endovascular aneurysm repair. [ABSTRACT FROM AUTHOR]

The present study was carried out to evaluate the effect of 5-aminolevulinic acid (ALA) on physiological and biochemical changes of in vitro-cultured Budaguvsky 9 (Bud 9) apple cultivar under NaCl induced salinity stress. Virus-free shoots and callus of Bud 9 apple were cultured on MS medium containing different levels of NaCl (0, 30, 60 and 90 mM) and ALA (0, 2.5, 5, 10 and 20 μM). Four weeks later phytochemical variations of regenerated plantlets with or without NaCl and/or ALA treatments were recorded. Shoot explants and callus treated with 5- aminolevolinic acid at 2.5 to 20 μM concentrations demonstrated reduction in catalase and peroxidase and enhancement in superoxide dismutase and ascorbate peroxidase enzyme activity. The highest chlorophyll a and b values were observed at shoot explants treated with concentration of 10 μm of aminolevolinic acid. Also, salinity stress was performed an effective role on increasing the amount of anthocyanins of explants and the highest amount was obtained in 90 mM salinity stress. Increasing the intensity of salinity stress and amino acid concentration (2.5-10 μm) in culture media had synergic effect on the antioxidant activity, total phenol as well as flavonoids content of studied explants. [ABSTRACT FROM AUTHOR]

An efficient protocol was developed to propagate old cypress of Abarkuh under in vitro condition. The explants from upper part of secondary shoots were successfully proliferated in the woody plant medium (WPM) supplemented with 30 g l−1 sucrose, 7.5 g l−1 agar, 0.5 mg l−1 benzyladenine (BA) and 0.01 mg l−1 indolebutyric acid (IBA). The in vitro raised micro shoots were then elongated by culturing in the WPM supplemented with 0.1 mg l−1 BA. Root induction was successfully conducted by pulse treatment (30 s) in high concentration of IBA (5 g l−1) and subsequent culturing in the WPM supplemented with 2 mg l−1 kinetin (Kin) and 0.01 mg l−1 IBA. Key message: An effective procedure was developed for successful in vitro micropropagation and plantlet regeneration of the cypress of Abarkuh, which is one of the oldest cypresses in the world with about 4000 years old. There were significant differences among basal media, BA concentrations as well as explant types on the proliferation of cypress of Abarkuh. The in vitro raised plantlets were successfully rooted and acclimated to the outdoor conditions. Tissue culture techniques can play an important role in multiplication as well as conservation of this valuable plant. [ABSTRACT FROM AUTHOR]

Background. Cellular therapy clinical applications require large-scale production of stem cells. Therefore, abundance, ease of isolation, and proliferative potential are the most important factors in choosing the appropriate source of cells for transplantation studies. Multipotent stem cells obtained from periodontal ligament (PDL) can be used in periodontal tissue regeneration. In this study, we aimed to evaluate and compare the characteristics of periodontal ligament stem cells (PDLSCs), extracted by either enzymatic digestion or explant methods, and expanded using two different serum types: fetal bovine serum (FBS) and xeno-free platelet lysate (PL). Methods. Expanded PDLSCs were assessed for their proliferation capacity, surface markers expression, colony formation, differentiation potential and ability to self- renewal. Most importantly, PDLSCs were evaluated for their ability to produce osteoblasts in vitro. Results. PDLSCs isolated by explant method and expanded in PL serve as a promising source of stem cells for osteoblasts regeneration. These cells showed higher proliferation capacity, they retained their stemness characteristics throughout the passages and they revealed an increase in the expression level of osteogenic markers, without showing any karyotypic abnormalities after cell expansion. Conclusions. PDLSCs produced using explant extraction method and expanded in cell culture media supplemented with PL provide an excellent source of xeno-free cells for the generation of functional osteoblasts. [ABSTRACT FROM AUTHOR]

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The analysis of the works of the scientists engaged in the Pseudotsuga menziesii propagation under the in vitro conditions has been made. The findings of the authors' researches on the propagation of the studied species by microcloning are presented. The optimal age of the Pseudotsuga menziesii maternal plants to procure initial plant material has been clarified. The explant decontamination (sterilization) schemehas been deduced from experiments, the nutrient medium compositionfor the in vitro initiation, multiplication and rooting of regenerated plants, as well as the substrate for the adaptation of the obtained clones to soil conditions, have been selected. The obtained results have shown the prospects of the studied species propagation by the proposed method. [ABSTRACT FROM AUTHOR]

Purpose: This study aims to analyze patient demographics, hospital characteristics, and clinical risk factors which predict penile prosthesis removal. We also examine costs of penile prosthesis removal and trends in inflatable versus non-inflatable penile prostheses implantation in the USA from 2003 to 2015. Methods: Cross-sectional analysis from Premier Perspective Database was completed using data from 2003 to 2015. We compared the relative proportion of inflatable versus non-inflatable penile prostheses implanted. We separated the prosthesis removal group based on indication for removal—Group 1 (infection), Group 2 (mechanical complication), and Group 3 (all explants). All groups were compared to a control group of patients with penile implants who were never subsequently explanted. Multivariate analysis was performed to analyze patient and hospital factors which predicted removal. Cost comparison was performed between the explant groups. Results: There were 5085 penile prostheses implanted with a stable relative proportion of inflatable versus non-inflatable prosthesis over the 13-year study period. There were 3317 explantations. Patient factors associated with prosthesis removal were non-black race, Charlson Comorbidity Index, diabetes, and HIV status. Hospital factors associated with removal included non-teaching status, hospital region, year of removal, and annual surgeon volume. Median hospitalization costs of all explantations were $10,878. Explantations due to infection cost $11,252 versus $8602 for mechanical complications. Conclusions: This large population-based study demonstrates a stable trend in inflatable versus non-inflatable prosthesis implantation. We also identify patient and hospital factors that predict penile prosthesis removal which has clinical utility for patient risk stratification and counseling. [ABSTRACT FROM AUTHOR]

The in vitro propagation of Colocasia esculenta is limited by the presence of microbial contaminants, especially bacteria. The objective of this work was to demonstrate that the inclusion of actions for the bacterial contamination management in the in vitro propagation of C. esculenta cv. 'INIVIT MC-2012', from field explants, can reduce losses. Two in vitro propagation protocols were used and the incidence of bacterial contaminants in the establishment and multiplication stages was compared. One protocol modified the other with actions for the management of bacterial contamination. In each one, 20 primary rhizomes were taken from which at least two axillary buds were extracted and from each one a line was established. The number of explants contaminated with bacteria per subculture was quantified. Bacterial contamination was observed in the culture medium below or around the explants, indicating the initial explant as the primary source of contamination. With the application of the modified protocol, the percentage of losses due to bacterial contamination in all subcultures evaluated was reduced. In the in vitro establishment it did not exceed 5%. The combination of actions for the management of bacterial contamination among which are the reduction of the size of the explant, the work by lines and the visual detection of contaminants prior to the subculture of the plant material reduces the presence of bacterial contaminants in the in vitro propagation of C. esculenta cv. 'INIVIT MC-2012'. [ABSTRACT FROM AUTHOR]

The paper presents the results of experiments performed for sugar beet (Beta vulgaris L.) plant regenerants obtaining of one sort and five hybrids through the indirect morphogenesis. Stages and peculiarities of regenerants' indirect morphogenesis in vitro culture, as well as specific characters of morphogenic structures in sugar beet calluses were studied. The effectiveness of triterpene saponosides usage as diagnostic markers, which determine potentially high productivity and adaptiveness of sugar beet (Beta vulgaris L.) plants, is proved. [ABSTRACT FROM AUTHOR]

The retina is the tissue responsible for light detection, in which retinal neurons convert light energy into electrical signals to be transported towards the visual cortex. Damage of retinal neurons leads to neuronal cell death and retinal pathologies, compromising visual acuity and eventually leading to irreversible blindness. Models of retinal neurodegeneration include 2D systems like cell lines, disassociated cultures and co‐cultures, and 3D models like organoids, organotypic retinal cultures and animal models. Of these, ex vivo human retinal cultures are arguably the most suitable models for translational research as they retain complex inter‐cellular interactions of the retina and precisely mimic in‐situ responses. In this review, we summarize the distinguishing features of the human retina which are important to preserve in experimental culture, the historical development of human retinal culture systems, the factors affecting ex vivo human retinal culture and the applications and challenges associated with current methods of human retinal explant culture. [ABSTRACT FROM AUTHOR]

One of most important issue in the field of regenerative medicine is selection of appropriate cells, scaffolds and bioreactors. The present study aimed to investigate the appropriate method for the isolation of human UC-MSCs cells from explant cultured in alginate scaffold within novel perfusion bioreactor. MSCs were isolated with explant method and CD markers such CD73, CD31, CD90 and CD105 as were analyzed by flow cytometry. The culture chamber of the novel perfusion bioreactor was made from Plexiglas and housed the cell/scaffold constructs in the central part and the medium for the whole culture period. The flow behavior within the bioreactor chamber were performed for closed and open bypass systems. The shear stress profiles simulated using CFD modeling. The fluid flow distribution within the bioreactor chamber was performed in PBS solution containing a blue colorant. UC explants were resuspended in sodium alginate and were allowed to polymerize and placed in the perfusion bioreactor and cultured. MSCs were positive for mesenchymal markers such as CD73 and CD31. All 3D Perfusion bioreactor parts, except peristaltic pump was sterilizable by autoclaving. Results of CFD indicated very low wall shear stress on surface of culture chamber at flow rate 2 ml/min. The maximum wall shear stress was 1.10 × 10-3 m/s = 0.0110 dyne/cm2 (1 Pa = 10 dyne/cm2). The fluid flow distribution within the alginate gel initially exhibited oscillation. In comparison, when encapsulated explants were placed in the perfusion bioreactor, cell proliferation appeared faster (4.6 × 1011 ± 9.2 × 1011) than explants cultures in 2D conventional culture method (3.2 × 1011 ± 1 × 1011). Proliferated cell formed several colonies. Migration of chondrocytes to the periphery of the alginate bead was visible after 1 week of culture. Perfusion bioreactor with low shear stress and alginate hydrogel improve cell isolation and expansion and eliminate cell passaging and enhance colony forming unit of UC-MSCs. [ABSTRACT FROM AUTHOR]

Abstract Meristem-tip of two important Iranian fig (Ficus carica L.) cultivars ('Jaami-e-Kan' and 'Sabz') were cultured under in vitro conditions to optimize the best condition for their shoot proliferation, root induction, and subsequent plantlet regeneration. Effects of explant size, culture medium (MS and B5) and different levels of BA (6-benzyladenine) in combination with 0.1 mg l−1 NAA (naphthalene acetic acid) were analyzed in the establishment of meristem cultures. The survival percentages of meristems were recorded after five weeks. Murashige and Skoog (MS) basal medium supplemented with different concentrations of BA (1, 1.5 and 2 mg l−1) and NAA (0, 0.1 and 0.5 mg l−1) was used for shoot regenerating. Half strength MS medium containing four concentrations of IBA (indole-3-butyric acid) was used for rooting. The regeneration and survival rate of cultures were significantly affected by different concentrations of BA, explant size and cultivars. The highest meristem survival was recorded from 'Sabz' cv., explants with the size of 0.5–0.7 mm, and culture media supplemented with 0.5 mg l−1 BA. The highest proliferation rate (2.4 shoots per micro shoot) was observed in the medium containing 2 mg l−1 BA, while the lowest length of shoots was obtained in this concentration. Furthermore, cv. 'Sabz' showed a higher proliferation rate (2.3 shoots per micro shoot) than cv. 'Jaami-e-Kan'. The highest numbers of rooted micro-shoots were obtained from 1.5 mg l−1 IBA (88.3%) followed by 2 mg l−1 IBA (78.5%). Also, 2 mg l−1 IBA showed the maximum number of roots per micro shoot (14.6). These data show that the presence of plant growth regulators, besides meristem size, plays an important role in meristem culture and micropropagation of fig trees. Moreover, results of this investigation can be applied practically for true to type as well as virus free plantlets regeneration from these important Iranian fig cultivars in a short time. Highlights • Meristem tip culture was successfully used for clonal multiplication of two commercial fig cultivars. • Responses of dry and freshly used figs were significantly different to the in vitro condition. • Meristem size plays an important role in the prosperity of meristem culture and micropropagation of fig trees. • Although fig is considered as a woody plant species, MS medium showed to be more suited than WPM for micropropagation of this fruit tree species. [ABSTRACT FROM AUTHOR]

Summary: Background: Laminitis has a considerable impact on the equine industry. Endocrinopathic laminitis is the most common form and affected horses often have hyperinsulinaemia due to an underlying metabolic disorder. Objectives: The aim of this study was to determine if insulin weakens the structural integrity of digital lamellae and to develop an ex vivo model for the study of hyperinsulinaemia‐induced lamellar failure. Study design: Ex vivo experiment. Methods: Biomechanical testing was used to assess the structural integrity of lamellar explants exposed to either medium alone (control) or medium supplemented with insulin. Lamellar explants comprised of hoof wall, lamellar tissue and distal phalanx were harvested from four adult horses with no evidence of inflammatory disease or pre‐existing disease of the digit. Following an equilibration period, explants were incubated in medium or medium supplemented with insulin (2.5 μg/ml) for 8 h prior to biomechanical testing to obtain load (N), stress (MPa), elongation to failure (mm), and Young's modulus (MPa) for each explant. Significant differences were assessed using a mixed linear model with horses as a random factor and control or insulin‐treated group as a fixed factor. Results: Lamellar explants incubated in medium supplemented with insulin failed at significantly lower load (P = 0.0001) and lower stress (P = 0.001) and had greater elongation to failure (P = 0.02). Main limitations: In addition to the ex vivo nature of the study, location‐dependent variability in explant structural integrity and variable diffusion of nutrients due to explant size may have been limitations. However, the study design attempted to account for these limitations through random assignment of explants to treatment groups independent of location and by evaluating stress to failure. Conclusions: Insulin weakens the structural integrity of equine lamellar explants and an ex vivo model for evaluation of hyperinsulinaemia‐induced lamellar failure was established. The summary is available in Spanish – see Supporting Information. [ABSTRACT FROM AUTHOR]

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

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

Differences in plant organs that are used as sources of explants can cause differences in callus growth and synthesized bioactive compounds. The objective of this study was to induce calli from different sources of explants and analyze their growth and bioactive compounds in the calli from leaf and stem explants of kaffir lime seedlings. Kaffir lime seeds were germinated until they grew into seedlings. On day 35, the leaves and stems of seedlings were harvested and callus was induced. Results showed that callus initiation time of stem explants was 5.66 days faster than that of leaf explants, which required 12.42 days. The colors of these calli were slightly different. Furthermore, callus fresh weight of leaf explants was less than that of stem explants, and the stationary phase of leaf explant-derived callus was earlier than for those from stem explant. Bioactive compounds detected in calli derived from leaf and stem explants were different. The main compounds found in the leaf explant-derived callus were n-decanoic acid and hexanedioic acid, bis (2-ethylhexyl) esters, while stem explant-derived callus had n-hexadecanoid acid. The presence of various bioactive compounds in these calli indicates potential for use as a natural medicine.

Introduction: German (Matricaria chamomilla L.) and Shirazi (Matricaria recutita L.) chamomiles are the most important medicinal plants of the Astraceae family which are used in the pharmaceutical, health, food and cosmetics industries. Production of this plant has been undertaken in Iran mainly in Isfahan, Kohgiluyeh and Boyerahmad, Golestan and Hamedan provinces. In vitro propagation of plants have higher potential to produce qualified natural products, restoring and preserving of endangered plants, induction of somaclonal variation, industrial reproduction, valuable secondary metabolites and increased active ingredients. Researchers reported successful micropropagation system for five chamomile varieties on MS medium contained 0.01 mg/l NAA and 2.5 mg/l kinetin. The aim of this study was to investigate the effects of stem (with and without node), leaf and cotyledon explants and different plant growth regulators on direct regeneration of German and Shirazi chamomiles. Although the Shirazi chamomile is native to Iran, but German chamomile is Iran non-indigenous cultivar. By our knowledge, there have been no comparison reports about responses of these cultivars to tissue culture. Materials and Methods: This research was performed in the central laboratory of Agriculture Faculty in Yasouj University. Seeds were provided from Pakan-Bazr institute, Isfahan. Chamomile seeds were disinfected by ethanol (70%) for 5-10 min and sodium hypocholorite 3% for 5-12 min and then washed for several times by distilled water. Then, seeds were sown on MS medium for germination. After 2-3 weeks, seedlings were grown and then planted in MS medium supplemented with hormonal combinations of NAA at two levels (0.1 and 0.5 mg/l), kinetin at three levels (2, 2.5 and 3 mg/l) and Zeatin, BAP and 2ip at three levels (0.5, 1 and 1.5 mg/l). The experiment was performed in a completely randomized design with four replications. Factors included explants, cultivars and hormonal combinations. In this experiment, traits such as stem induction percentage, stem length, stem fresh and dry weight root induction percent, root length and stem fresh and dry weight were measured. Statistical analysis was performed using SAS software (version 9.1). In order to test the normality and perform mean comparisons, Minitab 14 and MSTAT-C software was used and excel software was used for drawing diagrams. Results and Discussion: For all traits except stem induction percent and root dry weight, triple interaction of explants, hormonal combinations and cultivar were significant at 1% level. Triple mean comparisons for cultivar, explant and hormonal combinations shows that the highest mean for stem height, stem fresh weight, stem dry weight, root induction percent, root height and root fresh weight was obtained in Shirazi Chamomile cultivar with cotyledon explants in hormonal combinations of 0.1 mg/l NAA and 1.5 mg/l 2ip. The results showed that the best explants in both chamomiles for direct regeneration were stem (whit node) and cotyledon with 78.75% and 75% regenerations, respectively. Also the best genotype and hormone combination were Shirazi chamomile and MS medium supplemented with 1.5mg/l 2ip with 0.1mg/l NAA. To determine the best hormonal combination for root regeneration from direct regeneration, the stems were embedded in medium contained different concentrations of IBA. The first signs of rooting production were observed after 5-7 days. After completing the roots formation (4 weeks after transferring), the rooting percentage, root length, fresh and dry weight of roots were measured. Based upon variance analysis, effect of triple interactions of IBA, explants and cultivar on all traits were not significant, but the effect of IBA for all traits was significant at 1% level. According to the results of mean comparisons for effects of IBA on root traits in direct regeneration of chamomile, the highest percentage of root regeneration (73.75 %), the highest root length (6.60 cm), root fresh weight (174.167 mg) and the highest root dry weight (16.425 mg) were obtained from medium contained 0.5 mg/l IBA. Root differentiation was influenced by auxin (0.5 mg/l IBA) or spontaneously. Regenerated plantlets were transferred to pots contained sterilized soil (3:1:1 mixture of soil: sand: leaf compost). For plant adaptation to natural conditions, glassy caps were used. After adaptation, the caps were removed and the plants were transferred into a growth chamber. Previous studies reported that MS medium supplemented with 0.2 to 1 mg/l of BA and 2 mg/l of NAA induced adventitious bud formation and shoot development in leaf explants of Roman Chamomile. A higher number of adventitious buds were observed at the proximal end of the explants. Plantlets were rooted on MS medium supplemented with 0.1 mg/l of IBA and successfully weaned in vivo. Conclusion: Based on the results of this research, chamomile showed relevant response to direct regeneration.

Purpose. Developing technology for clonal micropropagation of peppermint (Mentha piperita L.) plants of Ukrainian breeding based on the complex of methods of isolated tissue and organ culture in vitro. Methods. During the experiment, such methods as isolated tissue and organ culture in vitro, clonal micropropagation, detached scion grafting, chemotherapy with adding of virucide Ribavirin to the nutrient medium, biometric and statistical ones were used. Results. The stepped procedure of sterilization that we have developed allows to receive 88–100% of sterile explants. For M. piperita L. introduction into culture and clonal micropropagation, Murashige and Skoog (MS) nut­rient medium appeared to be optimal supplemented with 6-benzylaminopurine (0.75 mg/l), adenine (0.05 mg/l), indole-3-acetic acid (IAA) (0.05 mg/l) and gibberellic acid (0.5 mg/l) on which the reproduction ratio on the 28th day ranged between 1:7 and 1:15. For recovery of plants from viral infection, virucide Ribavirin at concentration of 10 mg/l was added to the nutrient medium. The proposed nutrient medium for rhizogenesis, that contained IAA (0.5 mg/l) and indole butyric acid (IBA) (0.5 mg/l), allows to obtain the frequency of rhizogenesis up to 84–100%. Regenerated plants were adapted to the conditions in vivo on substrate peat : universal soil : perlite : sand in the ratio 2:1:1:1. The survival rate for peppermint varieties amounted to 96–100%. Conclusions. Biotechnological scheme was developed that permits to get healthy, purebred planting material and intensively propagate plants for supplying breeding programs of the Experimental Station for Medicinal Plants of the Institute of Agroecology and Environmental Management of the National Academy of Agrarian Sciences of Ukraine, among which such varieties as ‘Lebedyna pisnia’ and ‘Ukrainska pertseva’ were selected as the most promising for clonal micropropagation.

Scrophularia deserti is an important medicinal plant belonging to the Scrophulariaceae family. Research has demonstrated that extracts of the plant have antidiabetic and anticancer effects. This species is a valuable source of biologically active compounds, specially iridoid glycosides and flavonoids. Hairy root system can be used for the in vitro production of pharmaceutically valuable compounds. In this research, transgenic hairy root induction was established through the mediation of the A13 strain of Agrobacterium rhizogenes. The effects of various explants (seedling without radicle, cotyledon and leaf) prepared from in vitro grown plants at different ages (10, 18, 25, 50 and 72-day old) and three different inoculation times (5, 15 and 30 min) on the efficiency of hairy root induction were investigated. The maximum hairy roots (22.33 roots per explant) induced from 18-day-old seedlings explants inoculated with bacterial suspension for 5 min. The transgenic status of hairy roots was confirmed by PCR using rolA and rolB genes-specific primers. Four different culture media (MS, half strength MS, B5 and PGoB) were used to determine the best suitable media composition for the optimum cell growth of two hairy root lines (G and H). The results revealed that MS medium was superior for growth and high biomass production of hairy roots . Hairy root line H showed maximum biomass production (16.33 mg/30 ml culture medium, in terms of fresh weight) after 4 weeks of culture. Hairy root lines developed in this study can be used to investigate the production of pharmaceutically important metabolites of S. deserti.

Endemic plant species are usually more vulnerable to anthropogenic threats and natural changes and, therefore, hold a higher extinction risk. The preservation of these species is a major concern on a worldwide context and in situ protection alone will not guarantee their conservation. Ex situ conservation measures must be undertaken to support the conservation of these species, and seed banking is the more efficient and cost-effective method. However, when seed banking is not an option, alternative approaches should be considered. Biotechnological tools provide new and complementary options for plant conservation including short-, medium-, and long-term strategies, and their application for plant species conservation has increased considerably in the last years. This review provides information about the status of the use biotechnology-based techniques for the conservation of endemic plant species. Particular attention is given to cryopreservation, since is the only long-term ex situ conservation strategy that can complement and support the other conservation measures. The cryopreservation of plant genetic resources is, however, more focused on crop or economically important species and few studies are available for endemic plant species. The plant material used, the cryopreservation methods employed, and the assessment of cryogenic effects are reviewed. The reasons to explain the difficulties in cryopreserving these species are discussed and new strategies are proposed to facilitate and increase the interest on this matter. We expect that further studies on the conservation of endemic plant species will increase in a near future, thus contributing to maintain these valuable genetic resources.

The use of long-term mechanical assist devices for isolated right ventricular failure is rare. Herein, we describe our protocol for successful recovery of the right ventricle and subsequent explantation of a right ventricular assist device using a sternum-sparing technique and insertion of a titanium plug to occlude the coring defect of the inflow cannula. [ABSTRACT FROM AUTHOR]

Although plants of Vaccinium genus have not been cultivated on a large scale in the Czech Republic, there is a potential for commercial lingonberry production in some mountain regions. The purpose of this study was to develop an efficient in vitro system for a quick multiplication of lingonberry cvs Koralle, Linnea, and Runo Bielawskie. McCown woody plant medium (WPM), Anderson's rhododendron medium (AN) and half-strength Murashige and Skoog medium (half-MS) containing cytokinin zeatin in concentrations 0.5, 1 or 2 mg/l were tested for repeated subcultures. The number of newly formed shoots varied with the cultivar, medium tested and concentration of zeatin. Across all experiments, the highest multiplication rate (8.9 ± 0.6) was obtained for cv. Runo Bielawskie on WPM medium with the highest concentration 2 mg/l of zeatin. The lowest multiplication rates 1.1 ± 0.0 were noted on half-MS medium with the lowest concentration of zeatin (0.5 mg/l). In conclusion, micropropagation techniques described in this paper increased multiplication mainly in lingonberry cv. Runo Bielawskie on WPM medium. However, some cultivars of lingonberry would still require further research to optimize proliferation media.

Propagation of wild pistachio as a multipurpose woody species is a hard and tedious task. In this research, an effective in vitro protocol was developed for rapid proliferation of wild pistachio (Pistacia atlantica ssp. mutica) in MS medium supplemented with B5 vitamins and different growth regulators. Rooting of plantlets was tested by two treatments containing Rhizopon and IBA in ex vitro. With respect to the results, the nodal segments explants, produced the highest shoot frequency, leaf frequency and the tallest shoots. On the other hand, the tallest shoots were generated from shoot tip explant and medium containing of TDZ plus IAA. Both treatments (Rhizopon and IBA) led to a remarkable increase in the number of roots, root length and rooting percentage compared to the control. These results may be applied for rapid proliferation to spread the pistachio trees and shrubs that are difficult and time consuming.

Background: Elicitors are biotic or abiotic molecules that have effectively stimulated the production of plant secondary metabolites in plant, cell and organ cultures. Objective: Opium poppy serves as one of the particular commercial source of sanguinarine, as an anticancer and antimicrobial metabolite in the pharmaceutical industryThis study was done by the aim of increasing in expression of genes involve in the synthesis ofthisvaluablemetaboliteinplantcell suspension culture. Methods: In this study, the effects of two abiotic nano elicitors (nano TiO2 and nano-Silver), were studied on the expression of genes involve in sanguinarine biosynthesis pathway (tydc7, bbe1, DIOX2 and DBOX) at different time points (6, 24, 48, 72 and 168 hours) following treatments on shoot meristem and root suspension cell cultures using Real-time PCR. Results: All of these genes were expressedmost abundantly6 and 24 hours after treatment with nano elicitors in both root and meristem suspension cell cultures. After time all the genes expression were significantly decreased. Conclusion: Comparing the results of this study with the findings of other major studies on the effect of elicitors on papaver gene expression, indicated that because of the small sizes of nano particles, they may cause the rapid induction of sanguinarine biosynthesis genes expression in the shortest possible time. Nevertheless other investigations showed the highest increasing in gene expression, 48hours after elicitation. Given that, this study was conducted on callus with dedifferentiated cells from two different organs, but remarkable distinction were observed in gene expression profile in both explant tissues. In the other words the trace of cellular memory, was seen in dedifferentiated cells regards to the origin of explants.

Two accessions of potato (Glamočki and Rogatički) were used for the in vitro culture introduction in the Gene Bank of the Republic of Srpska. As explants for introduction are used germs of potato tubers. After surface sterilization, explants were inoculated on MS medium with 3% sucrose without hormones. Development of the explants was followed by five weeks and than were done selecting of survived explants. Number of survived explants was 70% for accessions of Glamočki potato, and 40% for Rogatički potato. Developed explants were used for next passage to period of 5 weeks and after that by micropropagation procedure transfered to MS medium with 3% sucrose and hormones BAP and IBA. After 5 weeks mesuring of lenth of shoots and determining of tubers number was performed. It was found that accessions of Glamočki potato had an average length of 12,24 cm of shoot while accessions of Rogatički potato had an average length 7,92. Average number of tubers per accession Glamočki potato had 1,81 while Rogatički had 1,21.

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

Failure by the developing conceptus to secrete sufficient interferon tau (IFNT), required for maternal recognition of pregnancy (MRP), at the appropriate time is related to early pregnancy loss in cattle. We aimed to test the hypothesis that there is a dose- and time-dependent relationship between IFNT and the endometrial expression of key interferon-stimulated genes (ISGs) involved in the signalling cascade leading to MRP in cattle. Candidate genes were identified first through a bioinformatic approach, where integrated transcriptomic data from two previous studies were analyzed to identify endometrial genes induced by IFNT. Next, expression of selected candidate genes was investigated in vitro in endometrial explants. Endometrial explants collected from cows (n = 8) in the late luteal phase of the estrous cycle were cultured in medium without (control) or with recombinant ovine IFNT (1, 10, 100 ng/mL) for 6 h. Simultaneously, endometrial explants were cultured in medium containing 100 ng/mL IFNT for different time periods (15 min, 30 min, 1 h, 3 h, 6 h). Gene expression was analyzed by RT-qPCR. We identified 54 endometrial genes responding to IFNT and to some degree to the conceptus, from which five ISGs (CMPK2, BPNT1, IFI35, TNFSF10 and TRIM38) were further selected for the dose- and time-dependent experiments. Classical ISGs (ISG15, OAS1, MX1 and MX2) were up-regulated (P < 0.05) in endometrium by 1 ng/mL IFNT. However, other selected ISGs (CMPK2, BPNT1, IFI35, TNFSF10 and TRIM38) were induced only by higher concentrations (10 and 100 ng/mL) of IFNT (P < 0.05). In terms of duration of exposure, IFNT at 100 ng/mL induced a significant (P < 0.05) increase in ISG15 and CMPK2 expression after 1 h incubation, while all other studied ISGs in the endometrium were upregulated when cultured for 3 or 6 h, but did not affect expression when the duration of culture was for 1 h or less. These results suggest that IFNT acts on the uterus in both a dose- and time-dependent manner in cattle and that timely exposure of the endometrium to sufficient IFNT is essential for appropriate signalling to ensure successful pregnancy establishment., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)