Karyological studies on the Sino-Himalayan genus, Cremanthodiurn (Asteraceae: Senecioneae)

Twenty-two populations of seven species of Cremanthodium from high altitude regions of western China were observed karyologically. C. ellisii, C. microglossum, C. brunneo-pilosum, C. stenoglossum, C. discoideum and C. lineare all had the same chromosome number of 2 n = 58 whereas C. humile had 2 n = 60. All chromosome numbers of these species are documented here for the first time. The basic number of x = 30 is new for this genus. The karyotypes of all species belong to 2A type according to Stebbins' asymmetry classification of karyotypes. Two basic chromosome numbers, x = 30 and x = 29 in Cremanthodium , correspond exactly to two branching patterns in this genus, sympodial versus monopodial. The systematic and taxonomic statuses of the sympodial species need further study. The karyomorphological data provide no support to the sectional subdivision in Cremanthodium .     

Ceramide Inhibits Cell Proliferation through Akt/PKB Inactivation and Decreases Melanin Synthesis in Mel‐Ab Cells

Ceramide is a bioactive sphingolipid that mediates a variety of cell functions. However, the effects of ceramide on cell growth and the melanogenesis of melanocytes are not known. In the present study, we investigated the actions of cell‐permeable ceramide and its possible role in the signaling pathway of a spontaneously immortalized mouse melanocyte cell line, Mel‐Ab. Our results show that C₂‐ceramide inhibits DNA synthesis in Mel‐Ab cells and G361 human melanoma cells in a dose‐dependent manner. Cell cycle analysis confirmed the inhibition of DNA synthesis by a reduction in the S phase. To investigate the ceramide signaling pathway, we studied whether C₂‐ceramide is able to influence extracellular signal‐regulated kinase (ERK) and/or Akt/protein kinase B (PKB) activation. We demonstrated that phosphorylated Akt/PKB is decreased by C₂‐ceramide, whereas phosphorylated ERK was only slightly affected. Therefore, the C₂‐ceramide‐induced inactivation of Akt/PKB may be closely related to the reduced cell proliferation of Mel‐Ab cells. Furthermore, we assessed the effects of C₂‐ceramide on the pigmentation of Mel‐Ab cells. The results obtained showed that the melanin content of cells was significantly reduced by C₂‐ceramide at concentrations in the range of 1–10 μM, and that the pigmentation‐inhibiting effect of C₂‐ceramide is much greater than that of kojic acid at 1–100 μM. In addition, we found that the activity of tyrosinase is reduced by C₂‐ceramide treatment. Our results demonstrate that C₂‐ceramide reduces the pigmentation of Mel‐Ab cells by inhibiting tyrosinase activity.     

Melatonin against acute ischaemic stroke dependently via suppressing both inflammatory and oxidative stress downstream signallings

This study tested the hypothesis that melatonin (Mel) therapy preserved the brain architectural and functional integrity against ischaemic stroke (IS) dependently through suppressing the inflammatory/oxidative stress downstream signalling pathways. Adult male B6 (n = 6 per each B6 group) and TLR4 knockout (ie TLR4^?/?) (n = 6 per each TLR4^?/? group) mice were categorized into sham control (SC^B6), SC^TLR4?/?, IS^B6, IS^TLR4?/?, IS^B6 + Mel (i.p. daily administration) and IS^TLR4?/? + Mel (i.p. daily administration). By day 28 after IS, the protein expressions of inflammatory (HMBG1/TLR2/TLR4/MAL/MyD88/RAM TRIF/TRAF6/IKK‐α/p‐NF‐κB/nuclear‐NF‐κB/nuclear‐IRF‐3&7/IL‐1β/IL‐6/TNF‐α/IFN‐γ) and oxidative stress (NOX‐1/NOX‐2/ASK1/p‐MKK4&7/p‐JNK/p‐c‐JUN) downstream pathways as well as mitochondrial‐damaged markers (cytosolic cytochrome C/cyclophilin D/SRP1/autophagy) were highest in group IS^B6, lowest in groups SC^B6 and SC^TLR4?/?, lower in group IS^TLR4?/? + Mel than in groups IS^TLR4?/? and IS^B6 + Mel and lower in group IS^B6 + Mel than in group IS^TLR4?/? (all P < .0001). The brain infarct volume, brain infarct area and the number of inflammatory cells in brain (CD14/F4‐88) and in circulation (MPO+//Ly6C+/CD11b+//Ly6G+/CD11b+) exhibited an identical pattern, whereas the neurological function displayed an opposite pattern of inflammatory protein expression among the six groups (all P < .0001). In conclusion, TLR inflammatory and oxidative stress signallings played crucial roles for brain damage and impaired neurological function after IS that were significantly reversed by Mel therapy.     

In Vitro Selection and Characterization of Human Immunodeficiency Virus Type 1 Variants with Increased Resistance to ABT-378, a Novel Protease Inhibitor

ABT-378, a new human immunodeficiency virus type 1 (HIV-1) protease inhibitor which is significantly more active than ritonavir in cell culture, is currently under investigation for the treatment of AIDS. Development of viral resistance to ABT-378 in vitro was studied by serial passage of HIV-1 (pNL4-3) in MT-4 cells. Selection of viral variants with increasing concentrations of ABT-378 revealed a sequential appearance of mutations in the protease gene: I84V-L10F-M46I-T91S-V32I-I47V. Further selection at a 3.0 μM inhibitor concentration resulted in an additional change at residue 47 (V47A), as well as reversion at residue 32 back to the wild-type sequence. The 50% effective concentration of ABT-378 against passaged virus containing these additional changes was 338-fold higher than that against wild-type virus. In addition to changes in the protease gene, sequence analysis of passaged virus revealed mutations in the p1/p6 (P₁′ residue Leu to Phe) and p7/p1 (P₂ residue Ala to Val) gag proteolytic processing sites. The p1/p6 mutation appeared in several clones derived from early passages and was present in all clones obtained from passage P11 (0.42 μM ABT-378) onward. The p7/p1 mutation appeared very late during the selection process and was strongly associated with the emergence of the additional change at residue 47 (V47A) and the reversion at residue 32 back to the wild-type sequence. Furthermore, this p7/p1 mutation was present in all clones obtained from passage P17 (3.0 μM ABT-378) onward and always occurred in conjunction with the p1/p6 mutation. Full-length molecular clones containing protease mutations observed very late during the selection process were constructed and found to be viable only in the presence of both the p7/p1 and p1/p6 cleavage-site mutations. This suggests that mutation of these gag proteolytic cleavage sites is required for the growth of highly resistant HIV-1 selected by ABT-378 and supports recent work demonstrating that mutations in the p7/p1/p6 region play an important role in conferring resistance to protease inhibitors (L. Doyon et al., J. Virol. 70:3763–3769, 1996; Y. M. Zhang et al., J. Virol. 71:6662–6670, 1997).     

Effects of Selected Chemical Pesticides on Agamermis unka (Nematoda: Mermithidae), a Parasite of the Brown Plant Hopper,Nilaparvata lugens

Selected commercial and technical grade pesticides were tested against the egg, preparasite and adult stages of Agamermis unka , a nematode parasite of the brown planthopper, Nilaparvata lugens . The commercial insecticide, diazinon (LC = 0.37 ppm), was most toxic to the 50 preparasites, followed by phenthoate (LC = 0.43 ppm), BPMC (LC = 0.44 ppm), IBP 50 50 (LC = 0.46 ppm), cartap hydrochloride (LC = 0.82 ppm) and buprofezin + isoprocarb 50 50 (LC = 1.11 ppm). The least toxic commercial pesticide tested was the fungicide, pencycuron 50 (LC = 2.19 ppm). Out of 12 technical grade insecticides tested, phenthoate, monocrotophos, 50 diazinon and carbofuran (LC = 0.37-0.46 ppm) were highly toxic to the preparasites, followed by 50 buprofezin, BPMC and fenitrothion (LC = 0.74-0.86 ppm). Fenthion, etofenprox, chlorpyrifos, 50 imidacloprid and MIPC (LC = 1.11-2.19 ppm) were the technical grade insecticides least toxic 50 to the preparasites. Most preparasites survived for up to 24 h at the low insecticide concentrations (0.63 and 0.31 ppm). Preparasites that were exposed to BPMC for 24 h at concentrations as high as 5.0 ppm and survived the treatments infected brown planthopper nymphs. Four selected insecticides-chlorpyrifos, BPMC, imidacloprid and carbofuran-had significant adverse effects on A. unka egg hatching. Eggs that were in the insecticide solution for 168 h fared poorly with imidacloprid having the best survival ( > 2% of the eggs hatching at 0.04 ppm). No eggs hatched from the other insecticide treatments. Three selected insecticides, BPMC, imidacloprid and chlorpyrifos, tested against adult A. unka showed that most adults survived the exposure to the insecticides between 0.31 and 2.5 ppm. At 5.0 ppm of BPMC or chlorpyrifos none of the adults survived, whereas with imidacloprid 70% of the adults survived. Egg deposition by the surviving adults was greatly reduced in those treated with the insecticides compared with those in the controls. Imidacloprid had some negative impact on the preparasites' ability to infect BPH nymphs, but it had the least detrimental effect of the insecticides tested on preparasite survival and on the eggs and adults of A. unka .     

Net Accumulation of Minerals and Water and the Carbon Budget in an Expanding Leaf of Tomato

The changes in f. wt, d. wt, nitrogen, potassium, calcium, phosphorus,magnesium and carbon of the seventh leaf of a tomato plant weremeasured at seven occasions from 10 days to 30 days after leafemergence. Measurements of CO₂ exchange by the leaf during bothlight (70 W m^–2, 7 h) and dark (17 h) periods and thechange in carbon content over these two periods enabled a carbonbalance to be constructed on these seven occasions. Changesin the sugars and starch contents of the leaf over these twoperiods at each occasion were measured. With the exception of calcium the rates of accumulation of allsubstances increased to their maxima when the leaf was 22–24-days-old.Carbon fixation per unit f. wt. increased to a maximum whenthe leaf was 16-days-old. In a 10-day-old leaf the rate of carbonfixation was already four-fifths of maximum and one-quartermore than that at 30 days. The rate of night respiratory lossof carbon per unit fresh weight decreased as the leaf expanded.In a 10-day-old leaf, the amount of carbon lost by night respirationaccounted for one-quarter of that fixed in the same day. Thisfraction fell to one-tenth when the leaf was 22-days-old andremained constant thereafter. The amount of carbon being importedto the leaflets of a 10-day-old leaf was less than one-quarterof that accumulated in 1 day. Thus, the contribution of theimported carbon to the leaf growth up to this stage is relativelysmall. The transition of the seventh leaf from being a net importerto being a net exporter occurred when the leaf was 13-days-old. The sucrose content per unit f. wt was higher in the youngerthan in the older leaves and was not correlated to the transitionfrom net import to net export. The accumulation and breakdownof starch in a leaf were related not only to the growth of theleaf but also to the development of the whole plant. Lycopersicon esculentum, tomato, leaf, accumulation of minerals, water content, carbon budget