Fractionation and Partial Characterization of Pectic Polysaccharides in Cell Walls from Liverwort (Marchantia polymorph) Cell Cultures

Konno, H., Yamasalu, Y. and Katoh, K. 1987. Fractionation andpartial characterization of pectic polysaccharides in cell wallsfrom liverwort (Marchantia polymorpha) cell cultures.—Jexp. Bot. 38: 711–722. Pectic polysaccharides were extracted from the starch-free cellwall preparation of cell suspension cultures of Marchantia polymorpha.The polysaccharides were fractionated by DEAE-Sephadex A-50ion-exchange chromatography yielding the five fractions, andthe degree of polymerization and glycosyl composition determinedfor each fraction. The neutral rich and acidic pectic polymerswere depolymerized by purified endoglucanase (l,4-ß-D-glucan4-glucanohydrolase, E.C. 3.2.1.4 [EC] .) and endopolygalacturonase(poly-l,4--Dgalacturonide glycanohydrolase, E.C. 3.2.1.15 [EC] ),respectively. The degraded pectic fractions were fractionatedby gel filtration chromatography on Bio-Gel A-5m and Bio-GelP-2, and glycosyl composition determined for each fraction.The results indicate that pectic polysaccharides contain glucose-richpolymer, rhamnogalacturonan and homogalacturonan in a ratioof 1:4:0–6. In addition, pectic polysaccharides were releasedas five pectic fragments from the cell walls by purified endopectatelyase (poly-l,4--D-galacturonide lyase, E.C. 4.2.2.2 [EC] ). Basedon the analysis of glycosyl composition of each fragment, thepectic polysaccharides of Marchantia cell walls are characterized Key words: Cell suspension culture, cell wall, liverwort, Marchantia polymorpha, pectic polysaccharides     

Up‐regulation of Tyrosinase Gene by Nitric Oxide in Human Melanocytes

Ultraviolet light (UV) radiation causes skin‐tanning, which is thought to be mediated by stimulating the release of melanogenic factors from keratinocytes as well as other cells. Nitric oxide (NO) has been reported to be generated after UV radiation and to stimulate melanocytes as one of the melanogens. In a previous experiment by another group on melanogenesis induced by NO, increases in both tyrosinase activity and tyrosinase protein levels were observed after daily stimulation of NO for 4 days. In the present study, we investigated tyrosinase gene expression within the first 24 hr of NO‐induced melanogenesis. Tyrosinase mRNA expression was found to be induced 2 hr after a single treatment with S‐nitroso‐N‐acetyl‐ l ‐arginine. An increase of tyrosinase activity was also detected time‐dependently within the 24‐hr period, accompanied by an increase of tyrosinase protein levels. The induction of mRNA expression was suppressed by a cyclic guanosine 3′,5′‐monophosphate (cGMP)‐dependent protein kinase (cGMP/PKG) inhibitor. These results suggest that the enhancement of tyrosinase gene expression via the cGMP pathway may be a primary mechanism for NO‐induced melanogenesis.     

A roundtable on responsible innovation with autologous stem cells in Australia,Japan and Singapore

We report on a roundtable event hosted in Singapore that sought to identify some of the ethical and regulatory challenges in translating autologous cell-based interventions, particularly those claiming to involve stem cells, into safe and effective therapies and to propose some solutions to encourage responsible innovation with these products. Challenges are identified in the three areas of cell manufacturing and processing, innovative uses of autologous cells in clinical practice and standards of evidence. Proposed solutions are discussed within a co-operative model of statutory laws and regulations that can enable product development with autologous cells and professional codes and standards that can encourage ethical conduct in clinical practice. Future research should be directed toward establishing regional networks for the development of internationally consistent standards in manufacturing and ethical codes of conduct for innovating with stem cells, and other autologous cells, and fostering ongoing exchange between jurisdictions.     

Determination of Macronuclear Fragmentation,Stabilization of Cell Union,and Cessation of Feeding Activity in Triplet Conjugants of Paramecium1

Triplet conjugants of Paramecium multimicronucleatum, in each of which the third cell unites by its anterior and to a posterior part of one of the conjugating pair, are induced by conjugation inducing chemicals, KCl and acriflavine in a Ca¹¹-poor condition. If the triplets are prematurely transferred to medium free of such chemicals, the third cells usually separate and no macronuclear fragmentation occurs in them. But in third cells of long lasting triplets, macromuclear fragmentation regularly begings at ～12 In after chemical treatment Staluhzaton of cell union and determmation of macronuclear fragmentation occured between ～100 and ～130 min after initiation of chemical treatment. Electron-microscopic observation of the stabilized third cell revealed that the two cell membranes at the uniting region were partially fused to form cytoplasmic bridges. Chemically treated cells began to cases feeding before the cell union and fully ceased such activity at ～130 min after the treatment.     

Involvement of Nitric Oxide in UVB‐Induced Pigmentation in Guinea Pig Skin

Ultraviolet (UV) B irradiation evokes erythema and delayed pigmentation in skin, where a variety of toxic and modulating events are known to be involved. Nitric oxide (NO) is generated from l ‐arginine by NO synthases (NOS). Production of NO is enhanced in response to UVB‐stimulation and has an important role in the development of erythema. NO has recently been demonstrated as a melanogen which stimulates melanocytes in vitro, however, no known in vivo data has been reported to support this finding. In this study, we investigated the contribution of NO with UV‐induced pigmentation in an animal model using an NOS inhibitor. UVB‐induced erythema in guinea pig skin was reduced when an NOS inhibitor, l ‐NAME (N‐nitro‐ l ‐arginine methylester hydrochloride), was topically applied to the skin daily, beginning 3 days before UVB‐irradiation. Delayed pigmentation and an increased number of DOPA‐positive melanocytes in the skin were markedly suppressed by sequential daily treatment with l ‐NAME. Furthermore, melanin content 13 days after UVB‐irradiation was significantly lower in skin treated with l ‐NAME than in the controls. In contrast, d ‐NAME (N‐nitro‐ d ‐arginine methylester hydrochloride), an ineffective isomer of l ‐NAME, demonstrated no effect on these UV‐induced skin responses. These results suggest that NO production may contribute to the regulation of UVB‐induced pigmentation.     

Microtubule Formation in Regenerating Terminal Buds of the Silurid Fish, Corydoras aeneus

Regenerating terminal buds of Corydoras aeneus were observed by electron microscopy to determine how terminal buds developed with respect to microtubule formation. After surgical removal of the fish barbel, it and the terminal bud began to regenerate 1.5 weeks later at 25°C. The regenerating terminal buds were ovoid in shape and contained three types of cells. The first type of cell had extended cellular processes which contained numerous microtubules and tubules. A bundle of three or four microtubules ran parallel to the long axis of the cellular process. Receptor villi protruded from the cell two weeks later, suggesting that it is a receptor cell. The second cell type, which appeared 1.5 weeks after barbel removal, had numerous microtubules oriented along the long axis of the cellular process; and numerous dense granules appeared two weeks later, suggesting that it is a supporting cell. The third type of cell observed was a basal cell without cellular processes. These results suggest that microtubule formation plays an important role in the elongation of regenerating terminal buds.     

ORAL CHEMORECEPTOR ORGANS OF BULLFROG TADPOLES DURING METAMORPHOSIS*

Degeneration of the premetamorphic papillae and development of the fungiform papillae during metamorphosis of bullfrog tadpoles were investigated by electrophysiological and scanning electron microscopic methods. Premetamorphic papillae were observed during the early metamorphic stages, and these degenerated rapidly at about metamorphic stage 20. The anlage of the tongue appeared at about metamorphic stage 10, but the anlage of the fungiform papillae appeared at about metamorphic stage 18. The microvilli at the apex of the fungiform papillae were observed at about metamorphic stage 21. At metamorphic stage 24 the fungiform papillae had a similar structure to that of adult frogs. Taste responses were recorded from the glossopharyngeal nerve of the tadpole. The responses to 1 M sucrose and 0.01 M quinine hydrochloride could be observed at metamorphic stage 6 or later, though during stage 20 the responses were very weak. The response to 0.02 M ammonium chloride appeared at metamorphic stage 6, but disappeared at stage 20 and did not reappear later. These results indicate that the fungiform papillae become functional as chemo-receptor organs at about metamorphic stage 21 and that, before the fungiform papillae function, the premetamorphic papillae serve as chemoreceptor organs in the tadpole.