Enigmatic thymus: Variations in anatomical localisation of thymic tissue as an easily misdiagnosed congenital anomaly in surgical practice

We point out the issue of differential diagnosis regarding the finding of ectopically localised thymic tissue(a thymic cyst)in the neck.Thymic tissue can be found anywhere along its developmental tract of descent,from the angle of the mandible to the upper mediastinum.Disruption of the thymic descent can result in ectopically/abnormally localised islets of accessory thymic tissue,which may undergo cystic changes,as described in a case report by Sun et al.This anatomical variation of the thymus may be clinically misinterpreted as a neoplasm or other congenital anomalies as a branchial cyst,lymphatic malformation or cystic hygroma.The present editorial focuses on the challenge of establishing a diagnosis of ectopically localised tissue of thymus often presented as a lateral cervical mass,especially in the case of cystic variation/degeneration of this thymic tissue.We summarise hypotheses on the origin of such congenital cervical thymic cysts from the point of view of evolutionary history and embryology.We also discuss lesser-known facts about the anatomy,histopathology and developmental biology of the thymus as one of the most enigmatic organs in the human body.

Plant growth-promoting properties and anti-fungal activity of endophytic bacterial strains isolated from Thymus altaicus and Salvia deserta in arid lands

Endophytes,as crucial components of plant microbial communities,significantly contribute to enhancing the absorption of nutrients such as nitrogen and phosphorus by their hosts,promote plant growth,and degrade pathogenic fungal mycelia.In this study,an experiment was conducted in August 2022 to explore the growth-promoting potential of endophytic bacterial strains isolated from two medical plant species,Thymus altaicus and Salvia deserta,using a series of screening media.Plant samples of Thymus altaicus and Salvia deserta were collected from Zhaosu County and Habahe County in Xinjiang Uygur Autonomous Region,China,in July 2021.Additionally,the inhibitory effects of endophytic bacterial strains on the four pathogenic fungi(Fusarium oxysporum,Fulvia fulva,Alternaria solani,and Valsa mali)were determined through the plate confrontation method.A total of 80 endophytic bacterial strains were isolated from Thymus altaicus,while a total of 60 endophytic bacterial strains were isolated from Salvia deserta.The endophytic bacterial strains from both Thymus altaicus and Salvia deserta exhibited plant growth-promoting properties.Specifically,the strains of Bacillus sp.TR002,Bacillus sp.TR005,Microbacterium sp.TSB5,and Rhodococcus sp.TR013 demonstrated strong cellulase-producing activity,siderophore-producing activity,phosphate solubilization activity,and nitrogen-fixing activity,respectively.Out of 140 endophytic bacterial strains isolated from Thymus altaicus and Salvia deserta,104 strains displayed anti-fungal activity against Fulvia fulva,Alternaria solani,Fusarium oxysporum,and Valsa mali.Furthermore,the strains of Bacillus sp.TR005,Bacillus sp.TS003,and Bacillus sp.TSB7 exhibited robust inhibition rates against all the four pathogenic fungi.In conclusion,the endophytic bacterial strains from Thymus altaicus and Salvia deserta possess both plant growth-promoting and anti-fungal properties,making them promising candidates for future development as growth-promoting agents and biocontrol tools for plant diseases.

Intrathyroidal ectopic thymus:Ultrasonographic features and differential diagnosis

Intrathyroidal ectopic thymus(IET)is defined as an ectopic thymus tissue that is generally found incidentally and rarely in the thyroid gland in the pediatric group.It occurs as a result of disruption of the embryological migration path and the settling of the thymus tissue into the thyroid gland.In the differential diagnosis,it is mostly confused with thyroid nodules.Although thyroid nodules are less common in children than adults,the rate of malignancy is much higher.Therefore,knowing the general ultrasound findings of IET better may prevent unnecessary invasive attempts and surgical procedures.In this article,we tried to compile the key imaging findings of IET.

The protection of Thymus vulgaris leaves alcoholic extract against hepatotoxicity of alcohol in rats

Objective:This study was performed to investigate the protective effect of Thymus vulgaris(T.vulgaris) leaves 70%alcoholic extract against alcohol-mediate hepatotoxicity rats.Methods:The protective effect of extract was investigated at dose of 500 mg/kg/day orally against alcohol-mediate hepatotoxicity using adult male Wister albino rats during 21 days.Protective effect of T.vulgaris extract was evaluated comparing with silymarin standard drug al recommended dose(25 mg/kg/day) orally for 21 days.Results:Alcohol-mediate hepatotoxicity rats(alcohol-control) showed hepatocytes distortion represented as marked increment on liver biomarkers;alkaline phosphatase(ALP),aspartate transaminase(AST) and alanine transaminase(ALT) activities,as well as pronounced reduction on total protein and its fractions albumin and globulin corresponding to normal ranges.Addition to oxidative stress status as depletion on glutathione concentration,catalase(CAT),superoxide dismutase(SOD),glutathione reductase(GR),glutathione-S-transferase(GST) and glutathione peroxidase(GPx)activities,concurrence with augmentation oxidative stress parameters;malondyaldchydc(MDA) and hydrogen peroxide(H_2O_2) concentrations comparing to normal values.Alcohol administration elevated total cholesterol(TC),low density lipoprotein cholesterol(LDL-C) and high density lipoprotein cholesterol(HDL-C) comparing to normal ranges.Co-administration T.vulgaris extract with alcohol showed protective effect on hepatocytes manifested as minimizing on ALP.AST and ALT activities and increment on total protein,albumin and globulin production compared to alcohol-control.Antioxidant enzymes activities;CAT.SOD.GR,GST and GPx were significantly magnified,while MDA and H_2O_2 concentration were lessened corresponding to alcohol-control.Also,lipid profile was markedly improved and risk ratio was lowered compared to alcohol-control.These results were confirmed by normalization of degenerated and fibrotic liver tissue as of alcohol-control.Conclusion:T.vulgaris extract appeared hepatoprotective,hypolipidemic and antioxidant activities on alcohol-mediate hepatotoxicity rats compared to silymarin.

Effect of Methionine Deficiency on the Thymus and the Subsets and Proliferation of Peripheral Blood T-Cell,and Serum IL-2 Contents in Broilers

The purpose of this study was to investigate the effects of methionine deficiency on cellular immune function by determining morphological and ultrastructural changes of thymus, thymic cell cycle and apoptosis, peripheral blood T-cell subsets, T- cell proliferation function and the serum interleukin-2 （IL-2） contents. 120 1-d-old broilers were randomly divided into two groups （6 replicates in each group and l0 broilers in each replicate） and fed on a control diet or methionine deficient diet for 42 d. Lesions were observed in experiment. Histopathologically, lymphopenia and congestion were observed in the medulla of thymic lobule. Ultrastructurally, there were more apoptosis lymphocytes, and the mitochondria of lymphocytes were swelled in thymus of methionine deficiency. The G0/G~ phase of the cell cycle of the thymus was much higher （P〈0.01）, and the S, G2＋M phases and proliferating index （PI） were lower （P〈0.01） in methionine deficiency than in control group. And the percentage of apoptotic cells in the thymus was significantly increased in methionine deficiency （P〈0.01）. The percentage of CD4＋ and CD8~ T-cells was decreased in methionine deficiency compared with control group. Meanwhile, the proliferation function of peripheral blood T-cell was decreased in methionine deficiency. Also, the serum IL-2 contents were decreased in methionine deficiency. It was concluded that methionine deficiency could cause pathological and ultrastructural changes of thymus, reduce the T-cell population, serum IL-2 contents and the proliferation function of T- cells, and induce increased percentage of apoptotic cells. The cellular immune function was finally impaired in broilers.

Gas chromatography-mass spectrometry analysis of different organic crude extracts from the local medicinal plant of Thymus vulgaris L

Objective:To isolate and analyze the chemical composition in different crude extracts of from the leaves of locally grown of Thymus vulgaris L(T.vulgaris)by gas chromatography-mass spectrometry(GC-MS).Methods:The shade dried leaves powder was extracted with methanol by using Soxhlet extractor.Methanol crude extracts of T.vulgaris and the derived fractions of hexane,chloroform,ethyl acetate and butanol were obtained.Results:Qualitative analyses of various organic crude extracts of T.vulgaris by using GC-MS showed that there were different types of high and low molecular weight compounds.Most of the isolated and identified compounds by GC-MS in the crude extracts are basically biologically important.Further,the T.vulgaris leaf possessed certain characteristics that can be ascribed to cultivation on a domestic plantation.The crude extracts were prepared from the powder leaves of T.vulgaris for respective compounds can be chosen on the basis of above GC-MS analysis.Conclusions:All the major compounds were identified and characterized by spectroscopic method in different organic crude extracts of T.vulgaris are biologically active molecules.Thus the identification of a good number of compounds in various crude extracts of T.vulgaris might have some ecological role.

Interactive effects of drought stress and chitosan application on physiological characteristics and essential oil yield of Thymus daenensis Celak

Thymus daenensis, a perennial herb, is often grown in areas that experience drought conditions during its growing period. Application of chitosan may compensate for the negative impact of drought stress on the yield of oil and secondary metabolites in Thymus.The interactive effects of foliar application of chitosan and drought stress on dry matter,essential oil yield, and selected physiological characteristics including photosynthetic pigments, osmotic adjustment, and lipid peroxidation of Thymus were investigated in a two-year study from 2014 to 2015. Treatments consisted of 0, 200, and 400(iL L'1 chitosan applied to plants grown under field capacity, mild drought stress(50% field capacity), and severe drought stress(25% field capacity). Dry matter yield decreased substantially as drought stress intensified. However, essential oil content increased under stress conditions,with the highest essential oil yield obtained from plants under mild drought stress. Foliar application of chitosan compensated to some extent for dry matter and oil yield reduction of plants grown under drought stress. The highest essential oil yield(1.52 g plant-1) was obtained by application of 400 \iL L_1 chitosan under the mild stress condition in 2015 when plants were mature. The compensatory effect of chitosan in reducing the negative impact of stress conditions on dry matter and oil yield was due mainly to stimulation of osmotic adjustment through proline accumulation and reduction of lipid peroxidase level, which increased the integrity of cell membranes of thyme leaves.

Antimycobacterial natural products from Moroccan medicinal plants: Chemical composition, bacteriostatic and bactericidal profile of Thymus satureioides and Mentha pulegium essential oils

Objective: To evaluate the susceptibility of Mycobacterium aurum and Mycobacterium smegmatis in vitro to the essential oils obtained from two medicinal plants: Thymus satureioides(T. satureioides) and Mentha pulegium(M. pulegium), and to study their chemical composition.Methods: The aerial parts of T. satureioides and M. pulegium(leaves and stems) were hydro-distillated using a Clevenger-type apparatus and essential oils were analyzed and identified by gas chromatography-mass spectrometry. Antimycobacterial screening of essential oils was performed on the basis of the inhibition zone diameter by disc diffusion method against two mycobacterial strains whereas the minimal inhibitory concentration and minimal bactericidal concentration were determined by using the micro-dilution method.Results: Chemical analysis of their aerial part's essential oils gave as major compounds,borneol(34.26%), carvacrol(31.21%) and thymol(3.71%) for T. satureioides and R(+)-pulegone(75.48%), carvone(6.66%) and dihydrocarvone(4.64%) for M. pulegium.Thereafter their antimycobacterial effect evaluation, using the micro-dilution method,indicated that minimal inhibitory concentration values of T. satureioides essential oil ranged from 0.062% to 0.015%(v/v) and from 0.125% to 0.031%(v/v) for M. pulegium respectively against Mycobacterium aurum and Mycobacterium smegmatis.Conclusions: It is clearly evident from the results obtained that the Moroccan medicinal plants have great potential to be used as anti-tuberculosis agents. These findings may help scientists to undertake several research projects to discover useful natural product as new anti-tuberculosis drug.

Antioxidant Machinery Related to Decreased MDA Generation by Thymus Algeriensis Essential Oil-induced Liver and Kidney Regeneration

Objective This study was conducted to determine the histopathological and biochemical effects of Thymus algeriensis essential oil （TEO） on hydrogen peroxide （H2O2）-induced oxidative stress in liver and kidney tissues of rats. Methods Rats were treated in six groups and were exposed for 2 weeks to low （LD; 100 μmol/L） and high doses （HD; 1 mmol/L） of H2O2 in the presence or absence of TEO （180 mg/kg）. Liver and kidney atrophy was measured by using biochemical and histopathological assays. Results Our study demonstrated that H2O2 induced liver and kidney atrophy, as evidenced by the significant elevation of serum aminotransferase, urea, and creatinine levels compared with those in the control rats. Urea levels were estimated by evaluating the activity of serum urease that hydrolyzes urea into CO2 and ammonia. However, TEO treatment significantly alleviated oxidative stress in the H2O2-induced liver and kidney toxicity model by reducing the levels of malondialdehyde concomitantly with marked elevations in superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase, as well as decrease in glutathione activity. Conclusion Our data demonstrated that TEO protected against H202 toxicity by decreasing oxidant levels and DNA damage, as well as increasing antioxidant levels, indicating that TEO has a spectrum of antioxidant and DNA-protective properties.

Study of total phenol,flavonoids contents and phytochemical screening of various leaves crude extracts of locally grown Thymus vulgaris

Objective:To prepare various crude extracts using different polarities of solvent and to quantitatively evaluate their total phenol,flavonoids contents and phytochemical screening of Thymus vulgaris collected from Al Jabal Al Akhdar,Nizwa.Sultanate of Oman.Methods:The leave sample was extracted with methanol and evaporated.Then it was defatted with water and extracted with different polarities organic solvents with increasing polarities.The prepare hexane,chloroform,ethyl acetate,hutanol and methanol crude extracts were used for their evaluation of total phenol,flavonoids contents and phytochemical screening study.The established conventional metliods were used for quantitative determination of total phenol,flavonoids contents and phytochemical screening.Results:Phytochemical screening for various crude extracts were tested and shown positive result for flavonoids,saponins and steroids compounds.The result for total phenol content was the highest in hutanol and the lowest in methanol crude extract whereas the total flavonoids contents was the highest in methanol and the lowest hexane crude extract.Conclusions:The crude extracts from locally grown Thymus vulgaris showed high concentration of flavonoids and it could be used as antibiotics for different curable and uncurable diseases.

The Interaction of Cobalt (Ⅲ) Mixed-Ligand Coordination Compound Containing Dipyrido[3,2-a:2′,3′-c] phenazine With Calf Thymus DNA -

The binding of Co (phen)2dppz3+ to calf thymus DNA,as investigated using absorption spectroscopy. viscosity and electrophoresis measurements,here dppz is dipyrido[3, 2-a: 2'3'-c]phenazine. The compound shows absorption hypochromicity and the specific viscosity increased upon binding to calf thymus DNA The complex is also shown to be more efficient photosensitisers for single strand breaks in plasmid DNA

Establishment and improvement of model of vascularized heart-thymus composite transplantation in rats

Objective To establish and improve the model of heart-thymus composite transplantation. Methods Vascularized both lobes of the thymus is transplanted heterotopically with the heart as a composite graft in rats.This technique was developed and assessed, and viability of the grafts was evaluated histologically. Results Donor operation costed 38. 5 ± 3. 52 min, vascular anastomosis costed 25.0 ± 3. 28 min, operating successful rate was 90%, acute rejection was observed in SD-Wistar group, viable thymus with normal microarchitecture was maintained in Wistar-Wistar group. Conclusions The improved novel technique for combined heart-thymus transplantation is a valuable method for study of the role of thymus in transplantation immunity.

Inhibitory effect of Thymus vulgaris extract on memory impairment induced by scopolamine in rat

Objective: To investigate the effect of Thymus vulgaris(T. vulgaris) on learning and memory functions in scopolamine-induced memory deficit in rats. Memory enhancing activity in scopolamine-induced amnesic rats was investigated by assessing the Morris water maze and passive avoidance paradigm.Methods: A total of 42 male Wistar rats were divided into 6 equal groups as follow:control group: received water, scopolamine treated group: received scopolamine 1 mg/kg for 15 days, two scopolamine + T. vulgaris treated groups: received scopolamine and T. vulgaris extract 50 and 100 mg/kg body weight per day for 15 days, two intact groups:received T. vulgaris extract 50 and 100 mg/kg body weight per day for 15 days.Results: Administration of T. vulgaris extract significantly restored memory and learning impairments induced by scopolamine in the passive avoidance test and Morris water maze test.Conclusions: T. vulgaris extract has repairing effects on memory and behavioral disorders produced by scopolamine and may have beneficial effects in the treatment of Alzheimer’s disease.

Chemical Composition, Antibacterial and Antioxidant Activities of Thyme Essential Oil (<i>Thymus vulgaris</i>)

Herbal medicine from natural resources plays an important role as antibacterial and antioxidant agents. The present investigation was designed to evaluate the antibacterial and antioxidant properties of thyme (Thymus vulgaris L.) essential oil (TEO) and/or chitosan (CH) in vitro. Results indicated that TEO exhibited high radical scavenging activity (RSA) toward DPPH, ABTS, linoleic acid deterioration and iron chelation activity. TEO exhibited high amount of total phenolic compounds (TPC) related to its terpenes. The TPC of TEO was 177.3 mg GAE g-1 demonstrated 149.8 μmol of TE g-1 DPPH-RSA and 192.4 μmol of TE g-1 ABTS-RSA. The antioxidant capacity of TEO exhibited 68.9% reduction when evaluated by β-carotene bleaching assay. The reducing power activity related to iron chelation was 142.8 μmol of AAE g-1. The TEO exhibited a high content of Thymol (41.04%) as major compound over 14 identified components by GC-MS analysis followed by 1,8-Cineole (14.26%), γ-Terpinene (12.06%), p-Cymene (10.50%) and α-Terpinene (9.22%). TEO exhibited antimicrobial activity in vitro and MIC noticed that TEO was efficiently affected pathogens in vitro. Indeed, CH exhibited negligible or very low antimicrobial activity. In conclusion, both investigated TEO and TEO-CHmix have strong antibacterial activity against many pathogenic bacteria and need exploitation as an alternative source of natural antibacterial and antioxidant agents for potential applications.

Chemical composition of essential oil of Thymus vulgaris collected from Saudi Arabian market

Objective: To focus on the analysis of chemical constituents of the Thymus vulgaris L.(locally known as "Zaitra" or "Za'atar"; Family: Lamiaceae) which is available in the market of Saudi Arabia.Methods: The Zaitra oil was analyzed by gas chromatography-mass spectrometry. The mass spectra were compared with the standard spectra available in National Institute Standard and Technique library.Results: The results indicated that the Thymus oil is composed of many chemical compounds including a pinene, thymol and caryophyllene which are biologically active and also used in various diseases.Conclusions: It can be concluded that the Thymus vulgaris due to presence of many bioactive compounds can be used as a new potential source of medicine for the treatment of various types of illness.

RUTHENIUM COORDINATION COMPOUNDS WITH POLYPYRIDYL：Ⅱ THE INTERACTION OF RUTHENIUM（Ⅱ）MIXED－LIGAND COORDINATION COMPOUNDS CONTAINING PYRIDO［3，2－f］［1，7］PHENANTHROLINE WITH CALF THYMUS DNA

The binding of Rulpdphen2+ to calf thymus DNA was inveatigated using absorption fluorescence and CD spectroscopies, where pdphen is the planar ligand pyrido[3. 2- f] [1, 7] phenanthroline,L=2, 2' -bipyridine, phenanthroline. 2, 9-dimethyl phenanthroline and 5-nitro-phenanthroline. All coordination componds show abaorption typochromicity associated with binding to calf thymus DNA .Ru(bipy) pdphen3+ and Rn (phen) pdphen+ also show fluoreacence intensities and excited state lifetime Increase becanse of the interaction of coordination compounds with calf thymus DNA .CD apectra of dialylatea indicate enantioselectivity associated withe binding to DNA .

Essential oil of <i>Thymus vulgaris</i>L. and <i>Rosmarinus officinalis</i>L.: Gas chromatography-mass spectrometry analysis, cytotoxicity and antioxidant properties and antibacterial activities against foodborne pathogens

The essential oil composition of Thymus vulgaris L. and Rosmarinus officinalis L. endemic to France were determined by GC and GC-MS. Oils were assessed for their cytotoxic, antioxidant and antimicrobial activity. 31 and 37 different compounds were identified representing 99.64% and 99.38% of the thyme and rosemary oils respectively, where oxygenated monoterpenes constituted the main chemical class. Thymol (41.33%) and 1.8-cineole (24.10%) were identified as the main constituents of T. vulgaris L. and R. officinalis L., respectively. Essential oils (EOs) of selected plant species were evaluated for their in vitro cytotoxicity against the human lung adenocarcinoma epithelial cell line (A549). Cytotoxicity was measured using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphynyltetra-zolium bromide] colorimetric assay. Dose-dependent studies revealed IC50 of 8.50 ± 0.01 μg/mL and 10.50 ± 0.01 μg/mL after 72 h on the A549 cells for R. officinalis L. and T. vulgaris L., respectively. Antioxidant activity was determined using a quantitative DPPH (1,1-diphenyl- 2-picryl hydrazyl) assay. Thymus and rosemary EOs exhibited effective radical scavenging capacity with 50% inhibitory concentration (IC50) of 437 ± 5.46 μg/mL and 189 ± 2.38 μg/mL respectively and therefore acts as a natural antioxidant agent. The antimicrobial activity of these species has also been studied against several foodborne pathogens and food isolated Salmonella spp. including S. enteritidis of significant importance. According to the results, T. vulgaris L. showed higher bactericidal effect than those from R. officinalis L. These results suggest that the essential oil from T. vulgaris L. and R. officinalis L. have potential to be used as a natural cytotoxic, antioxidant and antimicrobial agent in food processing.

Ruthenium Coordination Compounds with Polypyridines(Ⅵ)--Interaction of Ruthenium Polypyridine Complexes with Calf Thymus DNA-An Example of Non-intercalative Binding

Two new ruthenium polypyridine complexes, L 2RuL 2+ (L represents 2,2 bipyridine(bpy) or 1,10 phenanthroline(phen); L represents Schiff base derived from 4,5 diazafluoren 9 one(AFO) and n propylamine) have been prepared and characterized by means of elementary analyses, electronic absorption and IR spectra. The binding of the complexes to calf thymus DNA has been investigated via absorption, emission spectrometry, emission quenching, equilibrium dialysis and circular dichroism spectrometry. The experimental results show that these two complexes bind to calf thymus DNA by electrostatic mode and by non intercalative binding mode respectively.

Langerhans cell histiocytosis involving only the thymus in an adult: A case report

BACKGROUND Langerhans cell histiocytosis(LCH)is a rare disease of unknown etiology.LCH involving the thymus is mainly seen in pediatric patients and is extremely rare in adults.In this report,we describe a rare case of LCH originating from the thymus in an adult.CASE SUMMARY A 56-year-old man was admitted in April 2022 with complaints of intermittent dizziness since 2020,which had worsened in the previous 10 d.The physical chest examination was negative,and there was a history of hypertension for>2 years.Chest computed tomography showed a nodular soft tissue density shadow in the anterior mediastinum measuring approximately 13 mm×9 mm×8 mm.Postoperative pathological findings confirmed the diagnosis of LCH.CONCLUSION It is challenging to differentiate LCH involving the thymus from thymoma in imaging features.Pathological biopsy remains the gold standard when an anterior mediastinal occupying lesion is found.

Effect of Macronutrients, Cytokinins and Auxins, on <i>in Vitro</i>Organogenesis of <i>Thymus vulgaris</i>L.

The present study reports an efficient protocol for in vitro propagation of Thymus vulgaris L., an aromatic and medicinal plant in Morocco. Initially, we performed in vitro multiplication of Thymus vulgaris explants existing in the laboratory and obtained from micropropagation by shoot tip culture. Afterwards, we have evaluated the effect of six macronutrients. After that, seven cytokinins (Kin, BAP, 2iP, DPU, Adenine, Zeatine and TDZ) in three different concentrations (0.46, 0.93, 2.32 μM) have been evaluated to optimize cultures multiplication and elongation. Moreover, the effect of three auxins (IAA, IBA and NAA) at 0.57 μM, combined to 4 cytokinins (Kin, BAP, DPU and Ad.) at 0.46 μM, on shoot rooting has been studied. Thereby, MS medium has been proved the most favorable for plantlets growing. Also, we found that the addition of certain cytokinins, specifically 0.46 Kin, 0.46 and 0.93 BAP, 0.46 2iP, 0.46 DPU, 0.46 Ad. and 0.46 Zeat., ensures better multiplication and growth of vitroplants. In addition, multiplication and rooting of cultures were well optimized after addition 0.46 Kin + 0.57 IAA or NAA, 0.46 DPU + 0.57 IBA and 0.46 Ad. + 0.57 IBA combinations to the culture medium. Lastly, plantlets with roots were successfully acclimatized to ex-vitro conditions and these latter served as a source to establish in vitro culture again.