Cell and Tissue Journal

Abstract Aim: Artemisinin, an outstanding cumpound in genus Artemisia, is the most important anti malaria medicine. Therefore, plant cell culture of Artemisia aucheri Bioss was established and production of artemisinin was studied on plant, callus, and cell suspension culture.
Material and Methods: Artemisia aucheri aerial parts and seeds were obtained. Seedling was prepared and transferred on solid MS medium containing different growth regulators and callus was established. Fresh callus was transferred to liquid MS medium and suspension culture was obtained. For detection of artemisinin, the dichloromethanolic extract of callus, suspension and seeds of the plant was analysed by TLC and GC methods. Biotransformation was studied by feeding cholesterol, bisabolol and artemisia ketone to suspension culture.
Results: MS medium supplemented with kinetin (0.5 mg/l), 2, 4-D (0.5 mg/l), NAA (1 mg/l) and BA (0.25 mg/l), NAA (0.05 mg/l) was suitable for establishment of callus. Light showed positive effect on calli growth. No artemisinin was detected on plant aerial part. It seems, however, that callus and suspension culture of A. aucheri produced artemisinin. Also, Cholesterol, bisabolol and artemisinin keton feeding did not influence artemisinin production. Therefore, it seems these precursors are not proper for biotransformation experiments. 
Conclusion: Despite of undetected artemisinin in aerial parts of the A. aucheri, the production of artemisinin using new methods such as in vitro culture of A. aucheri is a promising result. 
 

Abstract Aim: In this study we were investigated the importance of the preparation method of the substrate based on PLGA polymer by electrospinning and freeze-drying method. To investigate the effect of nanotopogeraphy on cell behavior, the prepared nanofibers were compared with each other in three speeds of the collector.
Material & Methods: PLGA substratums were made by electrospinning and freeze-dying. The morphology of the structures was compared by use of the Scanning electron microscope images. Mouse fibroblast cells (L929 cell line) were seeded on substrates to determine the cell viability and followed by MTT assay.
Results: Scanning electron microscope images showed that, by increasing the speed of collector, the nanofibrous orientation increased. MTT assay (p < 0.05) showed in 24 h that PLGA substrates prepared by freeze-drying method were provided the appropriate attachment for the cell by producing a porous structure. Cell viability was significantly increased on the PLGA nanofibers with increase of fibers regularity after 48 and 72 h cell culture. This feature did not change on freeze-drying substrate.
Conclusion: The findings were indicated that electrospun PLGA nanofibers had the better performance to provide cell behavior. It was expected because of similarity to the structure of the natural extracellular matrix. It seems that aligned fibers acted as a positive factor to support cell proliferation.

Abstract Aim: The aim of this study was to evaluate the effects of rhizobial inoculation on the anatomical parameters of clover under SO₂ pollution.
Material and methods: 31 day-old plants (no-inoculated and inoculated with two strains of Rhizobium) exposed to the different concentrations of SO₂ (0 as a control, 0.5, 1, 1.5 and 2 ppm) for 5 consecutive days for 2 hours per day. The anatomical parameters of 40 day-old plants were Then investigated.
Results: Under high SO₂ concentrations (1, 1.5 and 2ppm), length of the palisade cells and diameter of the spongy cells decreased in both leaf surfaces. Stomatal density decreased in adaxial epidermis and increased in abaxial epidermis. Also, stomatal opening increased in adaxial epidermis and decreased in abaxial epidermis. Trichome density in both leaf surfaces significantly increased in 1.5 and 2ppm SO₂ compared with control plants. Under 2 ppm, trichome length increased significantly in both leaf surfaces. Inoculation of clover with two strains of Rhizobium significantly reduced the negative effects of high SO₂ concentration.
Conclusion: The results indicate the negative effects of high concentrations of SO₂ air pollution on the some anatomical parameters and the positive effects of bacterial inoculation on resistance to air SO₂ pollution stress.

Abstract Aim: Study of different factors affecting on improvement of growth and development of medicinal plants is very important. Inorganic elements are considered as those affecting factors. The aim of this research was to study the effects of different silicon concentrations on anatomical and developmental characteristics and some growth parameters of borage (Borago officinalis L.) in hydroponic conditions.
Material and Methods: The effects of different concentrations of silicon on anatomical features of epidermis and stomata cells and some growth parameters including quantity of photosynthetic pigments and proline in borage were studied. To this end, a hydroponics experiment was conducted randomly with different amounts of silicon including 0, 0.5, 1, 1.5, 2, 2.5 mM in four replications in greenhouse conditions.
Results: The results showed that 1.5 mM silicon resulted in a significant increase in stomata length and width, and stomatal index in borage leaf comparing with control and other treatments. Also application of 1.5 mM silicon showed positive effects on fresh weight of shoot and total chlorophyll content of plants compared with control. However it seems that silicon in high concentrations had negative effect on growth and anatomical attributes of the plant.
Conclusion: According to study of physiological and anatomical results, positive effects of silicon on pharmaceutical borage are just observed in appropriate and endurable concentrations for the plant.
 

Abstract Aim: Ethylene, a phytohormone, is produced during in vitro plant tissue culture and its accumulation is associated with reduction of growth and morphological changes. The aim of this study was to evaluate application of CoCl₂, as ethylene synthesis inhibitor, on growth and some physiological parameters as well as expression of ACC oxidase gene of potato.
Material and Methods: In this study, nodal segments of potato (Solanum tuberosum L.) cv. White Desiree were cultured on MS medium containing concentration of 0, 5, 10, 15, 20, 30, 40, 60 mg/L CoCl₂. After 5 weeks, chlorophyll a,b, total, cartenoied, content, leaf area and expression level of ACC oxidase gene were investigated.
Results: CoCl₂ inhibited root production in aerial parts and increased leaf areas and amount of chlorophyll a,b and carotenoid. Application of 20 mg/L CoCl₂ was the best for potato growth. ACC oxidase gene expression was decreased at this concentration.
Conclusion: Application of CoCl₂ decreased ACC oxidase gene expression and prevented ethylene accumulation in the in vitro culture and consequently improved growth of potato plant.

Abstract Aim: Previous studies using LOH technique on BDII rats showed four distinct Allelic imbalance on chromosome 15. In this research we have studied region 2 including Allelic imbalance by bioinformatic approachs and FISH (Florescence in situ hybridization) technique.
Material and Methods: The  confirmed tumors by an expert pathologist were used for cell culture. Metaphase chromosomes were prepared by conventional methods. The corresponding genes labeled probe for hybridization was poured onto slides. The labeled probe was then subjected to detected phase. Slides were studied under a fluorescence microscope using software CW4000 Laica.
Results: Using some of the database we could register 104 genes. But most of them did not have a clear function. According of FISH results, Dlgap5, Fermt2 and Socs4 had amplification and Lgals3 had copy number reduction.
Conclusion: according to our results, the Dlgap5 and Socs4 are probably involve in EAC

Abstract Aim: During recent years many studies have suggested some developmental roles for embryonic cerebrospinal fluid. Here we examined the effect of embryonic cerebrospinal fluid on proliferation and self-renewal of embryonic ventricular zone derived neurosphere.
Material and Methods: Cortex from 15.5 day old embryonic Wistar rat dissected out and enzymaticaly dissociated to form single cell suspension. Cell suspension seeded in DMEM/F12 medium supplemented with N2 and mitogenes (10ng.ml^-1 EGF and 20ng.ml^-1 bFGF). The CSF-treated culture from different embryonic ages (E16, E18, and E20) in 10/100 ratio (v/v) was considered as experimental groups. Neurospheres number was counted for proliferation assay and cell viability evaluated with MTT assay. The data were analyzed with One-way ANOVA with turkey’s post hoc test.
Results: Embryonic cerebrospinal fluid (e-CSF) enhanced neurosphere number. Also, embryonic cerebrospinal fluid (e-CSF) enhanced cell viability and growth.
Conclusion: Embryonic cerebrospinal fluid (e-CSF) enhanced proliferation and viability of neural progenitor cells in age dependent manner. Moreover, this fluid play important role in establishment of neuroprogenitor cells self-renewal.

Abstract Histological and Electron Microscopy Study of the Intestinal Tissue of Rainbow Trout (Oncorhynchusmykiss): an In Vitro Model

Abstract Aim: The aim of the present study was determination of FecB and FecG^H gene mutations in Kamouri goats of Khouzestan Province. Material and Methods: For this study 40 blood samples were collected of prolific kamouri goats. DNA of blood samples was extracted by modified salting out method and the site of the FecB and GDF9 genes were amplified using specific primers. Results: After amplification of the site of genes, the products of 190 bp and 139 bp were determined by agarose gel electrophoresis. The PCR products were digested with AvaII and DdeI enzymes. Results show no mutations in the FecB and FecG^H genes in the Khouzestan kamouri goats. Conclusion: In this study the mutations of FecB and FecG^H are not observed in Khouzestan kamouri goats and all of the goats were wild type. Therefore these mutations are not the general cause of the prolificacy in the Khouzestan Kamouri goats and further investigations are required to evaluating the fecundity genes and the genotyping of Khouzestan Kamuori goats.

Abstract Aim:The aim of this study was to investigate histopathological alterations and their incidence time pattern due to chromium (Cr) exposure during 18 days in gill, digestive gland and foot in freshwater bivalve, Anodonta cygnea.
Material and Methods: 24 bivalve specimens with length range of 12.7-13.3 cm were collected from Semeskandeh region, Sari. They were exposed to 125 µg l^-1 of Cr for 18 days. Bivalves were sampled in days 4, 9 and 18 for obtaining tissue samples of studied organs. Tissue sections of exposed and control group samples were prepared and staining was done using haematoxylin and eosin (H&E) method.
Results: Histology of all of three studied organs of exposed bivalves in comparison with control group showed significant histopathological changes. Hypoplasia and changes in shape and size of gill lamellae, granuloma, hyperplasia and atrophy of haemolymph channels were observed. In digestive gland, atrophy of digestive tubules, loss of digestive and basophilic cells from tubules, and haemocyte aggregation and granuloma in connective tissue were occurred. Hypoplasia of external epithelium, increase of mucus cells, and swelling and tissue rapture in myocyte blocks were observed in foot. Primary signs of histopathological alteration were appeared in fourth day, and in following days appearance of new signs and expansion of affected area was detected in all of investigated organs.
Conclusion: Based on our results, histological changes due to Cr exposure in studied organs in A. cygnea are proposed as appropriate histopathological biomarkers to monitoring Cr levels in aquatic environments and their incidence time pattern can increase fidelity of these indices.