ULTRASTRUCTURE OF THE VEGETATIVE CELLS OF NOSTOC FLAGELLIFORME PREPARED WITH HIGH PRESSURE FREEZING AND FREEZE SUBSTITUTION TECHNIQUE

The ultrastructure of the vegetative cells of Nostoc flagelliforme Born. et Flah. was investigated with high pressure freezing and freeze substitution technique and compared with the results obtained by using conventional preparation methods. During the processes of chemical fixation, dehydration and embedding, the cell structures might be more artificially modified than that obtained from high pressure freezing and freeze substitution. With the present method, the sheath of N. flagelliforme could be well penetrated and no extra big space could exist between the cell and the sheath. The cell protoplasm rarely shrinked. Some fine structures of cell inclusions and unit membranes became visualized. Many bacteria were harbored in the sheath. In addition, the presence of big vacuoles in the cell of N. flagelliforme as well as the presence of bacteria in the sheath shown in the present preparation for cyanobacteria has not been described so far in the literature.

Ultrastructure Features of Sperm Cell in Amaryllis vittata

Ultrastructure of sperm cells in pollen tube of Amaryllis vittata Ait. has been investigated in details by electron microscopy ,with particular emphasis on the organization and distribution of microtubules.The two newly formed sperm cells are arranged in tandem and sometimes in transverse at the right angle to the long axis of the pollen tube.Thevegetative nucleus is ahead and closely associated with the two sperm cells in all examined pollen tubes. The microtubules are distributed in the region between the common cell wall and the proximity of the sperm nucleus,they are singles and dispersed with mainly orientation of longitudinal and oblique,forming a loose bucket-like structure as a whole.In late stage of development,all of the microtubule array longitudinally and enclose the sperm nucleus.This configuration is very similar to the basket-like structure of microtubule in the generative cell. These results show that the organization and distribution of microtubules in the sperm cells are dynamic during cell development.

Effects of tachyplesin on the morphology and ultrastructure of human gastric carcinoma cell line BGC-823

AIM To investigate the morphological andultrastructural changes in the human gastriccarcinoma cell line BGC-823 after being treatedwith tachyplesin.METHODS Tachyplesin was isolated from acidextracts of Chinese horseshoe crab(Tachypleustridentatus)hemocytes.BGC-823 cells and thecells treated with 2.0mg/L tachyplesin wereexamined respectively under light microscope,scanning and transmission electron microscope.RESULTS BGC-823 cells had undergone therestorational alteration in morphology andultrastructure after tachyplesin treatment.Thechanges were as follows:the shape of cells wasunanimous,the volume enlarged and cellsturned to be flat and spread,the nucleo-cytoplasmic ratio lessened and nuclear shapebecame rather regular,the number of nucleolusreduced and its volume lessened,heter-chromatin decreased while euchromatinincreased in nucleus.In the cytoplasm,mitochondria grew in number with consistentstructure relatively,Golgi complex turned to betypical and well-developed,rough endoplasmicreticulum increased and polyribosomedecreased.The microvilli at cellular surfacewere rare and the filopodia reduced whilelamellipodia increased at the cell edge.CONCLUSION Tachyplesin could alter themalignant morphological and ultrastructuralcharacteristics of human gastric carcinoma cellseffectively and have a certain inducing differen-tiation effect on human gastric carcinoma cells.

Ultrastructure of neuronal-like cells differentiated from adult adipose-derived stromal cells

β-mercaptoethanol induces in vitro adult adipose-derived stromal cells （ADSCs） to differentiate into neurons. However, the ultrastructural features of the differentiated neuronal-like cells remain unknown. In the present study, inverted phase contrast microscopy was utilized to observe β-mercaptoethanol-induced differentiation of neuronal-like cells from human ADSCs, and immunocytochemistry and real-time polymerase chain reaction were employed to detect expression of a neural stem cells marker （nestin）, a neuronal marker （neuron-specific enolase）, and a glial marker （glial fibrillary acidic protein）. In addition, ultrastructure of neuronal-like cells was observed by transmission election microscopy. Results revealed highest expression rate of nestin and neuron-specific enolase at 3 and 5 hours following induced differentiation; cells in the 5-hour induction group exhibited a neuronal-specific structure, i.e., Nissl bodies. However, when induction solution was replaced by complete culture medium after 8-hour induction, the differentiated cells reverted to the fibroblast-like morphology from day 1. These results demonstrate that β-mercaptoethanol-induced ADSCs induced differentiation into neural stem cells, followed by morphology of neuronal-like cells. However, this differentiation state was not stable.

Ultrastructure of human neural stem/progenitor cells and neurospheres

BACKGROUND： Biological and morphological characteristics of neural stem/progenitor cells （NSPCs） have been widely investigated. OBJECTIVE： To explore the ultrastructure of human embryo-derived NSPCs and neurospheres cultivated in vitro using electron microscopy. DESIGN, TIME AND SETTING： A cell biology experiment was performed at the Brain Tumor Laboratory of Soochow University, and Jiangsu Province Key Laboratory of Neuroregeneration, Nantong University between August 2007 and April 2008. MATERIALS： Human fetal brain tissue was obtained from an 8-week-old aborted fetus; serum-free Dulbecco＇s modified Eagle＇s medium/F12 culture medium was provided by Gibco, USA; scanning electron microscope was provided by Hitachi Instruments, Japan; transmission electron microscope was provided by JEOL, Japan. METHODS： NSPCs were isolated from human fetal brain tissue and cultivated in serum-free Dulbecco＇s modified Eagle＇s medium/F12 culture medium. Cells were passaged every 5-7 days. After three passages, NSPCs were harvested and used for ultrastructural examination. MAIN OUTCOME MEASURES： Ultrastructural examination of human NSPCs and adjacent cells in neurospheres. RESULTS： Individual NSPCs were visible as spherical morphologies with rough surfaces under scanning electron microscope. Generally, they had large nuclei and little cytoplasm. Nuclei were frequently globular with large amounts of euchromatin and a small quantity of heterochromatin, and most NSPCs had only one nucleolus. The Golgi apparatus and endoplasmic reticulum were underdeveloped; however, autophagosomes were clearly visible. The neurospheres were made up of NSPCs and non-fixiform material inside. Between adjacent cells and at the cytoplasmic surface of apposed plasma membranes, there were vesicle-like structures. Some membrane boundaries with high permeabilities were observed between some contiguous NSPCs in neurospheres, possibly attributable to plasmalemmal fusion between adjacent cells. CONCLUSION： A large number of autophagosomes were observed in NSPCs and gap junctions were visible between adjacent NSPCs.

Ultrastructure Changes and Mechanism of Cancer Cells Induced by Inorganic Crystal Nanoparticles

The purpose of this study was to examine the changes of cancer cell in ultrastructure after inorganic crystal nanoparticles （ICN） absorption. HAP and TiO2 nanoparticles were incubated with the Bet- 7402 cells for 1 h and 8 h respectively. Then, cancer cells were collected and examined under transmission electron microscope （TEM）. In cytoplasm, nanoparticle were contained in some vacuoles. Some death features of cell appear. The experimental results indicated that ICN can induce cancer cells death.

Lycopene modulates cellular proliferation, glycolysis and hepatic ultrastructure during hepatocellular carcinoma

AIMTo investigate the effect of lycopene extracted from tomatoes (LycT) on ultrastructure, glycolytic enzymes, cell proliferation markers and hypoxia during N-Nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis. METHODSFemale BALB/c mice were randomly divided into four groups: The Control, NDEA (200 mg NDEA/kg b.w. given i.p.), LycT (5 mg/kg b.w. given orally on alternate days) and LycT + NDEA group. The mRNA and protein expression of various cell proliferation markers (PCNA, Cyclin D1, and p21) were assessed by reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay, respectively. The ultrastructure of hepatic tissue was analyzed using scanning and transmission electron microscopy. The enzymatic activity of glycolytic enzymes was estimated using standardized protocols, while glucose-6-phosphate dehydrogenase activity level was estimated using a kit obtained from Reckon Diagnostic P. Ltd. (India). RESULTSUncontrolled proliferation in the liver of NDEA (P ≤ 0.001) mice was evident from the high expression of cell-proliferation associated genes (PCNA, Cyclin D1, and p21) when compared to control and LycT mice. In addition, enhanced activities of hexokinase, phosphoglucoisomerase, aldolase, glucose-6-phosphate dehydrogenase and hypoxia-inducible factor-1α were observed in NDEA mice as compared to control (P ≤ 0.001) and LycT (P ≤ 0.001) mice. The alterations in hepatic ultrastructure observed in the NDEA group correlated with the changes in the above parameters. LycT pre-treatment in NDEA-challenged mice ameliorated the investigated pathways disrupted by NDEA treatment. Moreover, hepatic electron micrographs from the LycT + NDEA group showed increased macrophages, apoptotic bodies and well-differentiated hepatocellular carcinoma (HCC) in comparison to undifferentiated HCC as observed in the NDEA treated group. CONCLUSIONThis study demonstrates that dietary supplementation with LycT has a multidimensional role in preventing HCC development.

Changes in Cell Ultrastructure in Maize Leaves Infected by Maize Dwarf Mosaic Virus

Ultrastructural alterations in foliar cells were studied in leaves of resistant maize variety Luyu16 and susceptible maize inbred line Luyuan92 infected by maize dwarf mosaic virus Shandong isolate (MDMV-SD), respectively. The results showed that marked cytopathological alterations were observed both in resistant plants and in susceptible plants, compared with that in healthy plants. However, some ultrastructural alterations, which observed in resistant plants, were different from those in susceptible plants. In resistant plants, which infected with the virus, the main organelles, including chloroplasts and mitochondria, were slightly destroyed, the amount of mitochondria and peroxisome were increased. A few or no plasmodesmata were observed. There were three kinds of inclusions including pinwheel, bundle and laminated aggregate, and the virus particles in the cytoplasm. In susceptible plants, which infected with the virus, the chloroplasts were heavily disrupted, including thylakoid swelling and envelope broking. The virus particles were more than those in the resistant variety. Four kinds of inclusions including pinwheel, bundle, laminated aggregate and high electon-dense body appeared in cytoplasm. Plasmodesmata and plasma membrane were abundant, and there were frequent invaginations of the plasma membrane that led to the formation of vesicles and myelin-like structures.

THE EFFECT OF ACTIVE COMPONENTS OF LYCIUM BARBARUM AND GARLIC (LB-GO) ON THE SYNTHESIS OF DNA AND ULTRASTRUCTURE OF U_(14) CERVIX CANCER CELLS IN MICE

Mice, inoculated with U_(14) ascitic type cervix cancer cells, were administered with effective components of Lycium barbarum (LB) and garlic (GO).On the 4th day after i.p.administration, general condition of the animals were found improved.Examination of ascitic fluid revealed damage of the cancer cells, blanching of fluorescence staining of DNA and RNA, and the cancer cells besieged by large numbers of macrophages and leucocytes. Flow cell metric (FCM) analysis found: accumulation of cells of G_1 stage. Ultrastructure study disclosed: swelling of mitochondria in cytoplasm and damage of mitochondrial crests even with cavity formation, enlargement and degranulation of rough ER. It seemed the effect of LBGO was affirmable. It was postulated that macrophages,being activated by LB, came in close contact with the cancer cells giving rise to carcinolysis. In addition to the direct but transient killing effect of GO, the anti-cancer results could be greatly enhanced.

The Association of Chalazal Cell Ultrastructure in Caryopsis and Grain Filling in Wheat

Using three wheat cultivars (Triticum aestivum L. ) , Een1, Huamai8 and 95A-10, which exhibit different grain filling characters. the relationship of ultrastructure of the chalazal cells in wheat caryop-sis and grain filling was studied. The results indicate that the lipoid and polysaccharide deposited in vacuoles of the chalazal cells have no obvious inhibitory actions to the grain filling. It appears that a direct relationship exists between the proceeding and end time of grain filling and the amount and appearing time of the lipoid, within lignin and suberin deposited on the inner wall of chalazal cells, and between grain filling rate and cultivars characteristic. The appearing time of the lipoid, within lignin and suberin deposited on the inner wall of chalazal cells in cultivar with large grain (Eenl and 95A-10) was later than that in cultivar with small grain (Huamai8), and the amount of deposits in the former was less than that in the latter. The late development of caryopsis and low rate of grain filling account for the wrinkled grain of 95A-10. The transport of assimilates in chalazal cells coexists with symplastic and apoplastic pathways, but symplastic pathway plays a major role after 24 days of flowering.

Ultrastructure of Single Cells, Callus-like and Monospore-like Cells in Porphyra yezoensis Ueda on Semi-solid Culture Medium

It had been demonstrated that individual cells or protoplasts isolated fromPorphyrathallus by enzyme could develop into normal leafy thalli in the same way as monospores, and that isolated cells develop in different way in liquid and on semi-solid media. The authors observed the ultrastructure of isolated vegetative cells cultured on semi-solid media and compared them with those of monospores and isolated cells cultured in liquid media. The results showed that subcellular structures were quite different among cells in different conditions. In their development, isolated cells on semi-solid media did not show the characteristic subcellular feature of monospore formation, such as production of fibrous vesicles. Callus-like cells formed on semi-solid media underwent a distinctive modification in cellular organization. They developed characteristic cell inclusions and a special 2-layer cell covering. Golgi bodies, ER, starch grains, mitochondria. Vacuoles were not commonly found in them.

Effects of hydrogen peroxide on ultrastructure and cellular permeability of Candida albicans

The changes of the ultrastructure and the cellular permeability of Candida albicans were ob-served after the fungus was exposed to 3%～10% hydrogen peroxide.Damages on the fungus cellsespecially cytoplasm leakage were found under transmission electron microscopy and morphologicalchanges with small hole-like structures under scanning dectron microcopy.Radioactivity assay ofthe leakage of [~3H]-TdR-DNA of C.albicans further proved the existence of cellular permeabilitydamage.There was negative correlation between the amount of [~3H-TdR-DNA leakage and thenumber of the survival fungus cells,which indicates that the damage on cellular permeability andDNA leakage caused by hydrogen peroxide of certain concentration is one of the important factorsto kill the fungus.

THE ULTRASTRUCTURE OF SEPARATED AND CULTURED CELL OF PORPHYRA YEZOENSIS

There are many reports that cells (protoplasts) separated from the thallus of Porphyra by enzyme can develop to normal leafy thalli in the same way as monospores. But there are few investigations on the subcellular structure of the isolated vegetative cell for comparison with the subcellular structure of monospores. To clarify whether the separated and cultured cells undergo the same or similar ultrastructure changes during culture and germination as monospores undergo in their formation and germination, we observed their ultrastructure, compared them with those of the monospore and found that the ultrastructure of separated and cultured cells did not have the characteristic feature as that of monospore formation, such as production of small and large fibrous vesicles, but was accompanied by vacuolation and starch mobilization like that in monospore germination. The paper also discusses the relations between monospores and separated and cultured cells.

STUDY ON THE ULTRASTRUCTURE OF SINGLE HeLa CELL TREATED WITH HpD PLUS LASER MICROIRRADIATION

A method is described which permits transmission electron microscope of single cells treated with HpD plus laser microirradiation. The preselected single cell that was irradiated by laser under light microscope and followed fixation, embedded and sectioning is examined under electron microscope. The results demonstrated that at the light dose of 1.88 ml/μm2 not only the irradiated nucleolus appeared transparent region, but the other parts such as non-irradiated mitochondria in cytoplasm can also be damaged. When partial cytoplasm is irradiated with the light dose of 4.50 ml/μm2, the damages appear in all cytoplasm, but there is little change in the nucleus. The experimental results also demonstrate that cytoplasm is more sensitive than nucleus. It is the mitochondria in cytoplasm that are very sensitive to HpD plus laser.

Ultrastructure of oval cells in children with chronic hepatitis B,with special emphasis on the stage of liver fibrosis:The first pediatric study

AIM:To investigate the ultrastructure of oval cells in children with chronic hepatitis B,with special emphasis on their location in areas of collagen fibroplasia. METHODS:Morphological investigations were conducted on biopsy material obtained from 40 children,aged 3-16 years with chronic hepatitis B. The stage of fibrosis was assessed histologically using the arbitrary semiquantitative numerical scoring system proposed by Ishak et al. The material for ultrastructural investigation was fixed in glutaraldehyde and paraformaldehyde and processed for transmission-electron microscopic analysis. RESULTS:Ultrastructural examination of biopsy specimens obtained from children with chronic hepatitis B showed the presence of two types of oval cells,the hepatic progenitor cells and intermediate hepatic-like cells. These cells were present in the parenchyma and were seen most commonly in areas of intense periportal fibrosis (at least stage 2 according to Ishak et al) and in the vicinity of the limiting plate of the lobule. The activated nonparenchymal hepatic cells,i.e. transformed hepatic stellate cells and Kupffer cells were seen in close proximity to the intermediate hepatic-like cells. CONCLUSION:We found a distinct relationship between the prevalence of oval cells (hepatic progenitor cells and intermediate hepatocyte-like cells) and fibrosis stage in pediatric patients with chronic hepatitis B.

The Study on Culture and Ultrastructure of Glaucomatous Trabecular Cells in Vitro

Purpose : To establish the culture system of human glaucomatous trabecular cells in vitro and study their ultrastructures.Methods : The trabecular specimens from trabeculectomy were cultured in vitro and passaged 3 times, then identified. Moreover, the glaucomatous cells were observed with electron microscope while compared with the normal ones.Result: Cultured human glaucomatous trabecular cells were obtained. The ultrastructure of the cells showed the decrease in vilious project, coated vesicle and lysosomal inclusion. Conclusion : The establishment of human glaucomatous trabecular cells culture in vitro made the culture system more perfect. The morphologic changes might be related to the abnormal functions of human trabecular meshwork cells. Eye Science 1998; 14 : 134 - 737.

Seasonal Changes in the Ultrastructure of the Vascular Cambium in Shoots of Populus tomentosa

Wood is the product of cambial activity in trees, and the seasonal activity style of cambium directly influences wood biomass production, structures and properties. The seasonal changes in the ultrastructure of the vascular cambium activity of Populus tonientosa Carr. planted in Beijing area were examined in shoot tissues collected during 15 months by means of transmission electron microscopy. Before xylem mother cells reactivated completely, the dividing fusiform cells in cambium and new phloem cells had appeared at the same time. The initiation of cambial activity may be related to the bud sprouting and the young leaf growth in shoots. More details about the ultrastructural changes of cambial cells at the onset of cambial activity have been gained. When the large vacuole in active cambial cells divided into smaller ones during the dormant phase, proteinaceous material that disappeared in active cambial cells refilled many of these small vactioles. In addition, lipid droplets and starch granules had the same cycles as proteinaceous material. The plasmalemma invaginations of fusiform cells were observed not only in active phase but also in dormancy. The endomembrane system consisting of nuclear membrane, endoplasmic reticulum (ER), dictyosomes and their secretory vesicles, changed in form and distribution at different phases during a cycle and performed important roles at the onset of active cambium and during the wall formation process of secondary xylem cells. The tangential walls remained relatively thin throughout the year but the radial walls thickened markedly when the cambium was dormant. During the transition from dormancy to activity, a partial autolysis occurred in the radial walls of the cambial cells, especially at the cell wall junctions. A notable feature of the cells at the onset of cambial activity was the thinning of the radial walls.

Characters of Fractal Ultrastructure in Wood Cell Wall

Fractal theory was introduced in order to describe the ultrastructure of wood cell wall in this paper. The cellulose chain clusters around nano-scale were viewed as a fractal object that consists of many fibrillar structural units with different scales including microfibrils. On the basis of the morphological data of wood cell wall, fractal dimensions of multi-level fibrillar structural units were calculated by fractal-geometry approach, and then the morphological and structural characteristics of fibers as well as the influences on wood properties were investigated according to the dimensions. Besides, the fractal self-nesting character of the ultrastruture was also analyzed.

Pectin methylesterase inhibitors GhPMEI53 and AtPMEI19 improve seed germination by modulating cell wall plasticity in cotton and Arabidopsis

The germination process of seeds is influenced by the interplay between two opposing factors,pectin methylesterase(PME)and pectin methylesterase inhibitor(PMEI),which collectively regulate patterns of pectin methylesterification.Despite the recognized importance of pectin methylesterification in seed germination,the specific mechanisms that govern this process remain unclear.In this study,we demonstrated that the overexpression of GhPMEI53is associated with a decrease in PME activity and an increase in pectin methylesterification.This leads to seed cell wall softening,which positively regulates cotton seed germination.AtPMEI19,the homologue in Arabidopsis thaliana,plays a similar role in seed germination to GhPMEI53,indicating a conserved function and mechanism of PMEI in seed germination regulation.Further studies revealed that GhPMEI53 and AtPMEI19 directly contribute to promoting radicle protrusion and seed germination by inducing cell wall softening and reducing mechanical strength.Additionally,the pathways of abscicic acid(ABA)and gibberellin(GA)in the transgenic materials showed significant changes,suggesting that GhPMEI53/AtPMEI19-mediated pectin methylesterification serves as a regulatory signal for the related phytohormones involved in seed germination.In summary,GhPMEI53 and its homologs alter the mechanical properties of cell walls,which influence the mechanical resistance of the endosperm or testa.Moreover,they impact cellular phytohormone pathways(e.g.,ABA and GA)to regulate seed germination.These findings enhance our understanding of pectin methylesterification in cellular morphological dynamics and signaling transduction,and contribute to a more comprehensive understanding of the PME/PMEI gene superfamily in plants.