Effects of Ultraviolet Irradiation on the Cell Cycle

Cultured mouse Cloudman melanoma cells, EMT6 breast carcinoma cells, and 3T3 fibroblasts all accumulated in the G2/M phase of the cell cycle when exposed to UVB radiation. The effects of UVB were maximal at 20–30 mJ/cm² for all three cell lines, and could be observed by flow cytometry as early as 12 hr post irradiation. It has been known since the mid-1970s that MSH receptor binding activity is highest on Cloudman melanoma cells when they are in the G2/M phase of their cycle. Here we show that either UVB irradiation or synchronization of Cloudman cells with colchicine results in a stimulation of MSH binding within 24 hr following treatment, a time when both treatments have resulted in accumulation of cells in the G2/M phase of the cycle. Furthermore, the two treatments performed together on the melanoma cells stimulated MSH receptor activity to the same extent as either treatment performed separately, suggesting that each may be influencing MSH receptor activity solely through a G2/M accumulation of cells. Together, these results raise the possibility that an increase in the number of cells in the G2 phase of the cell cycle is a generalized cellular response to injury, such as UV irradiation. However, in the case of pigment cells this response includes a mechanism for increasing melanin formation, i.e., increased MSH receptor activity. Should this be the case, similar G2/M “injury responses” of other cell types might be expected, consistent with their differentiated phenotypes.     

Molecular Cascades in UV Induced Melanogenesis: A Central Role for Melanotropins?

When human skin is exposed to ultraviolet (UV) light, a highly complex cascade of events ensues that culminates, among other things, in increased skin melanin content. From analyses at the tissue and cellular level, it has been shown that following exposure to UV light there is an increase in the number of active melanocytes in the basal layer of the epidermis, and individual melanocytes are stimulated to produce more melanin. In addition, the rate of transfer of melanosomes from melanocytes to keratinocytes is apparently increased, although the role of UV light in this process remains to be demonstrated. Recent biochemical evidence is reviewed on factors that regulate these processes. A plausible explanation for the effects of UV on pigmentation is that there are mechanisms in the skin for the orderly, regulated reception of UV signals that are then transduced to initiate the cascade. The signals involve both melanocytes and keratinocytes, and avail-able evidence supports a model in which melanotropins and their receptors play a central role in the process.     

Detection of cross reactive antigens between Pestalotiopsis theae and tea leaves and their cellular location

Among the 12 varieties of tea tested against three isolates of Pestalotiopsis theae, causal agent of grey blight disease, Teen Ali-17/1/54 and TV-23 were found to be highly susceptible while CP-1 and TV-26 were resistant under identical conditions. Leaf antigens were prepared from all the tea varieties, three isolates of P. theae and a non-pathogen of tea (Bipolaris tetramera). Polyclonal antisera were raised against mycelial suspensions of P. theae (isolate Pt-2) and leaf antigens of Teen Ali-17/1/54 and CP-1. These were compared an immunodiffusion test and enzyme-linked immunosorbent assay to detect cross reactive antigens (CRA) shared the host and the parasite. CRA were found among the susceptible varieties and isolates of P. theae (Pt-1, 2 and 3). Such antigens were not detected between isolates of P. theae and resistant varieties, B. tetramera and tea varieties or isolates of P. theae. Indirect staining of antibodies using fluorescein isothiocyanate (FITC) indicated that in cross sections of tea leaves, the CRA was concentrated in the epidermal cells and mesophyll tissues. CRA was present in the young hyphal tips of the mycelia and on the setulae and appendages of the conidia of P. theae.     

Effect of Arbutin on Melanogenic Proteins in Human Melanocytes

The inhibitory effect of arbutin, a naturally occurring β-D-glucopyranoside derivative of hydroquinone, on melanogenesis was studied biochemically by using human melano-cytes in culture. Cells were cultured in the presence of different concentrations of arbutin. The maximum concentration of arbutin that was not inhibitory to growth of the cells was 100 ug/ml. At that concentration, melanin synthesis was inhibited significantly by ～20% after 5 days, compared with untreated cells. This phenotypic change was associated with the inhibition of tyrosinase and DHICA polymerase activities, and the degree of inhibition was dose dependent. No significant difference in DOPAchrome tautomerase (DT) activity was observed before or after arbutin treatment. Western blotting experiments revealed there were no changes in protein content or in molecular size of tyrosinase, TRP-1 or TRP-2, indicating that inhibition of tyrosinase activity by arbutin might be due to effects at the post-translational level.     

Melanoma × Macrophage Fusion Hybrids Acquire Increased Melanogenesis and Metastatic Potential: Altered N-Glycosylation as an Underlying Mechanism

We recently reported that a majority of hybrids generated in vitro between weakly metastatic mouse Cloudman S91 melanoma cells and human or mouse macrophages showed enhanced metastatic potential. With few exceptions, hybrids with enhanced metastatic potential also had elevated basal melanin content and increased responsiveness to MSH compared to parental cells. Here we investigated the hybrid melanotic phenotype in more detail, comparing the pigmentary systems of hybrids and parental Cloudman S91 cells by several techniques. Cells were studied by electron microscopy, cell lysates were analyzed for tyrosinase (E.C.1.14.18.1) activity, and melanosomal proteins were analyzed by gel electrophoresis and immunoblotting. Melanosomes in parental Cloudman melanoma cells were few in number and relatively amorphous, whereas those in the hybrids were numerous and heavily pigmented, containing highly organized lattice structures. Both basal and MSH-inducible tyrosinase activities were elevated several fold in hybrids compared to parental cells. Tyrosinase, TRP-2, and LAMP-1 from hybrids migrated more slowly on gels compared to the same proteins from parental melanoma cells, consistent with increased glycosylation. Migration of LAMP-1 from hybrids was similar to that from peritoneal macrophages, which also appeared to be more heavily glycosylated than LAMP-1 from Cloudman cells. By using ³H-glucosamine as a marker of N-glycosylation, its incorporation into tyrosinase and LAMP-1 was found to be elevated in hybrids, suppressed by N-glycosylation inhibitors, and stimulated by MSH to a greater degree in hybrids compared to parental cells. These results indicate N-glycosylation as an important regulatory pathway for MSH-induced melanogenesis and further suggest that altered N-linked glycosylation may be an underlying mechanism for regulation of both melanogenesis and metastasis in macrophage x melanoma hybrids.     

Electron Microscopic Studies on the Basal Apparatus of the Flagellum in the Protozoon, Leishmania donovani

SYNOPSIS. The basal apparatus of the flagella and kinetoplast in Leishmania donovani have been studied with the electron microscope. The flagellar fibrils extend into the body of the protozoan to form the kinetosome. At the point of origin of the flagellum, the pellicle invaginates to form a kinetosomal vacuole around the kinetosome. The kinetoplast is formed by a transversely elongated banded structure, surrounded at some distance by a double layered kinetoplast membrane. There is no apparent connection between the kinetosome and the kinetoplast.     

印度Mahanadi盆地井下二叠纪下冈瓦纳岩层的微体植物化石组合

Mahanadi河盆地的冈瓦纳型诸盆地位于印度的近中心地区,呈北西—东南走向分布。Mahanadi主要盆地内的几个重要煤田为Korda煤田,Mand—Raigarh煤田和Ib河煤田。Korda煤田的冈瓦纳系沉积向南东向延伸,经Mand—Raigarh煤田延续至Ib河煤田。Korda和Ib河两个煤田储量巨大,且长期来有小构造活动;而Mand—Raigarh煤田至今很少受到重视,勘探开发晚得多。文中首次对Mand-Raigarh煤田5个钻孔样品作了孢粉研究,以初现面、相对丰度等为特征,划分出7个孢粉组合带,时代从二叠纪最早期(Talchir组)至二叠纪末期(Raniga由组)。填补了东西两个煤田之间的生物地层研究空白。7个孢粉组合带为：i)Parasaccites-Plicatipollenites assemblage zone(相当于Talchir组）,ii)Sulcatisporites-Brevitriletes assemblage zone(相当于下Barakar组）,iii)Sulcatisporites-Rhizomaspora assemblage zone(少量Densipollenites存在说明其相当于上Barakar组）,iv)Densipollenites-Striatopodocarpites assemblage zone(相当于Barren Measures组）,v)Striatopodocarpites-Densipollenites assemblage zone(相当于下Raniganj组）,vi)Striatopodocarpites-Densipollenites-Sulcatisporites assemblage zone(相当于上Raniganj组）vii)Sriatopodocarpites-Alisporites assemblage zone(相当于Raniganj最上部到Kamthi组最下部）,Raniganj组沉积中见到小的皱缩裂隙、植物垂直根和根的印痕,似代表古土壤。     

Electron Microscopy of the Hypotrich Ciliate Oxytricha platystoma, with Consideration of its Macronuclear Organization

SYNOPSIS. The body of Oxytricha platystoma is covered with 2 membranes, the outer or pellicular membrane and the inner or cytoplasmic membrane. Each cirrus consists of a bundle of 25-50 cilia. There are 2 macronuclei and 2 micronuclei. The micronucleus is a very small body containing dense Feulgen-positive material. The macronucleus is an elongated body surrounded by a double membrane. The dense Feulgen-positive material is embedded in a less dense nucleoplasm. Some larger bodies found inside the nucleus may be nucleoli. There is a single reorganization band in each macronucleus. It consists of a solution plane and a reconstruction plane, and has a total thickness of 1.5 μ. The macronucleus divides amitotically, whereas the micronucleus divides mitotically.     

Production and fitness of Fusarium pseudograminearum inoculum at elevated carbon dioxide in FACE

Rising atmospheric carbon dioxide (CO₂) concentration is increasingly affecting food production but how plant diseases will influence production and quality of food under rising CO₂ is not well understood. With increased plant biomass at high CO₂ the stubble‐borne fungal pathogen Fusarium pseudograminearum causing crown rot (CR) of wheat may become more severe. We have studied inoculum production by Fusarium using fungal biomass per unit wheat stubble, stem browning from CR and the saprophytic fitness of Fusarium strains isolated from two wheat varieties grown in 2007 and 2008 at ambient and elevated CO₂ in free‐air CO₂ enrichment (FACE) with or without irrigation and once in a controlled environment. Fungal biomass, determined using primers for fungal ribosomal 18s and the TRI5 gene, increased significantly at elevated CO₂ in two of the three studies. Stem browning increased significantly at elevated CO₂ in the 2007 FACE study. At elevated CO₂ increased stem browning was not influenced by irrigation in a susceptible variety but in a resistant variety stem browning increased by 68% without irrigation. Wheat variety was significant in regression models explaining stem browning and Fusarium biomass but pathogen biomass at the two CO₂ levels was not significantly linked to stem browning. Fusarium isolates from ambient and elevated CO₂ did not differ significantly in their saprophytic fitness measured by the rate of colonization of wheat straw. We show that under elevated CO₂Fusarium inoculum in stubbles will be amplified from increased crop and pathogen biomass while unimpeded saprophytic fitness will retain its effectiveness. If resistant varieties cannot completely stop infection, Fusarium will rapidly colonize stubble to further increase inoculum once the crop is harvested. Research should move beyond documenting the influence of elevated CO₂ to developing disease management strategies from improved knowledge of pathogen biology and host resistance under rising CO₂.