alpha-Tomatine, the major saponin in tomato, induces programmed cell death mediated by reactive oxygen species in the fungal pathogen Fusarium oxysporum

The tomato saponin alpha-tomatine has been proposed to kill sensitive cells by binding to cell membranes followed by leakage of cell components. However, details of the modes of action of the compound on fungal cells are poorly understood. In the present study, mechanisms involved in alpha-tomatine-induced cell death of fungi were examined using a filamentous pathogenic fungus Fusarium oxysporum. alpha-Tomatine-induced cell death of F. oxysporum (TICDF) occurred only under aerobic conditions and was blocked by the mitochondrial F(0)F(1)-ATPase inhibitor oligomycin, the caspase inhibitor D-VAD-fmk, and protein synthesis inhibitor cycloheximide. Fungal cells exposed to alpha-tomatine showed TUNEL-positive nuclei, depolarization of transmembrane potential of mitochondria, and reactive oxygen species (ROS) accumulation. These results suggest that TICDF occurs through a programmed cell death process in which mitochondria play a pivotal role. Pharmacological studies using inhibitors suggest that alpha-tomatine activates phosphotyrosine kinase and monomeric G-protein signaling pathways leading to Ca(2+) elevation and ROS burst in F. oxysporum cells.     

Flavoring and extending the shelf life of cucumber juice with aroma compounds-rich herbal extracts at 4 °C through controlling chemical and microbial fluctuations

This work aims to enhance the flavor of functional cucumber juice using herbal extracts of peppermint, basil, lavender, and lemongrass ethanolic extracts and extend its lifetime by controlling the chemical and microbial fluctuations. Cucumber juices were processed as; non-supplemented (J-Con), J-PME, J-BE, J-LE, and J-LEE supplemented with peppermint, basil, lavender, and lemongrass ethanolic extracts, respectively. Peppermint extract was significantly scavenged 88% of DPPH radicals and inhibited the growth of tested gram-positive, gram-negative bacteria and fungi followed by the lemongrass extract. The antioxidant activity of cucumber juices increased due to polyphenols and aroma compounds in the added extracts. However, the antioxidant content was decreased after two months of storage at 4 °C, due to the decrease in polyphenols. The flavor compounds were determined using GC mass, wherein hydrocarbons, acids, alcohols, and carbonyl compounds were the main aroma contents in cucumber juices, and their contents decreased with storage time. Peppermint and lemongrass extracts were significantly (p ≤ 0.05) increased the whiteness of J-PME, and J-LEE, respectively. The highest score of flavor and taste was observed in J-PME that scored 8.3 based on panelists'' reports followed by J-LEE. The PME was significantly maintained 91% of the odor and color of J-PME as compared to other juices.     

Kaempferol triosides from Reseda muricata

A flavonoid trioside and its coumaryl ester together with seven known flavonoids and five phenolic acids were isolated from the leaves of Reseda muricata. Their structures were elucidated by spectroscopic methods including UV, FAB MS, 1H, 13C and 2D-NMR, DEPT, HMBC and HMQC experiments. The two compounds were identified as kaempferol 3-O-beta-D-glucopyranosyl-(1' --> 2')-O-alpha-L-rhamnopyranoside 7-O-beta-D-glucopyranoside and kaempferol 3-O-beta-D-glucopyranosyl-(1' --> 2')-O-alpha-L rhamnopyranoside 7-O-beta-D-(6'-O-E-coumarylglucopyranoside), respectively.     

The ingi and RIME non-LTR retrotransposons are not randomly distributed in the genome of Trypanosoma brucei

The ingi (long and autonomous) and RIME (short and nonautonomous) non--long-terminal repeat retrotransposons are the most abundant mobile elements characterized to date in the genome of the African trypanosome Trypanosoma brucei. These retrotransposons were thought to be randomly distributed, but a detailed and comprehensive analysis of their genomic distribution had not been performed until now. To address this question, we analyzed the ingi/RIME sequences and flanking sequences from the ongoing T. brucei genome sequencing project (TREU927/4 strain). Among the 81 ingi/RIME elements analyzed, 60% are complete, and 7% of the ingi elements (approximately 15 copies per haploid genome) appear to encode for their own transposition. The size of the direct repeat flanking the ingi/RIME retrotransposons is conserved (i.e., 12-bp), and a strong 11-bp consensus pattern precedes the 5'-direct repeat. The presence of a consensus pattern upstream of the retroelements was confirmed by the analysis of the base occurrence in 294 GSS containing 5'-adjacent ingi/RIME sequences. The conserved sequence is present upstream of ingis and RIMEs, suggesting that ingi-encoded enzymatic activities are used for retrotransposition of RIMEs, which are short nonautonomous retroelements. In conclusion, the ingi and RIME retroelements are not randomly distributed in the genome of T. brucei and are preceded by a conserved sequence, which may be the recognition site of the ingi-encoded endonuclease.     

Alkaloid accumulation in Catharanthus roseus cell suspension cultures fed with stemmadenine

Feeding stemmadenine to Catharanthus roseus cell suspension culture resulted in the accumulation of catharanthine, tabersonine and condylocarpine. Condylocarpine is not an intermediate in the pathway to catharanthine or tabersonine when it is fed to the cultures. The results support the hypothesis that stemmadenine is an intermediate in the pathway to catharanthine and tabersonine.     

Growth, metabolic profiling and enzymes activities of Catharanthus roseus seedlings treated with plant growth regulators

The effect of different growth regulators on growth and the production of terpenoid indole alkaloids as well as some enzymes involved in the biosynthesis were studied in Catharanthus roseus seedlings. The seedlings were grown on MS solid medium containing different concentrations of each growth regulator for a period of one month. Extracted alkaloids were analyzed by HPLC for determination of terpenoid indole alkaloid quantities. Continuous availability of growth regulators induced different alkaloids with variable effects among the regulators. Gibberellic acid at concentration of either 5.8 M or 11.6 M resulted in elongation of shoots with lowering the number of leaves. Abscisic acid has a retardant effect on growth. Ethylene did not effect the growth pattern at concentration of 100 M but seedlings were not tolerant to higher concentrations. Methyljasmonate reduced the growth of the root system. Methyljasmonate was a general inducer for all alkaloids and increased the activity of strictosidine glucosidase. Ethylene applications promoted the pathways towards ajmalicine, serpentine, tabersonine and vindoline. Similar effect as for ethylene was observed for abscisic acid. Salicylic acid treatment increased the production of serpentine, tabersonine and higher concentration of salicylic acid induced vindoline accumulation. Peroxidase activity was also induced by salicylic acid. Gibberellic acid has little effect on alkaloid levels.     

Synthesis of aldehydo-sugar derivatives of pyrazoloquinoline as inhibitors of herpes simplex virus type 1 replication

Synthesis of a novel series of structurally related pyrazoloquinoline nucleosides is described. All the newly synthesized compounds were examined for their in vitro antiviral activity against herpes simplex type-1 as shown by two different bioassays, namely; crystal violet staining or the MTS tetrazolium dye measurement. The acute toxicity (LD50) values of the biologically active compounds were determined.     

Proton transfer reactions in native and deionized bacteriorhodopsin upon delipidation and monomerization

We have investigated the role of the native lipids on bacteriorhodopsin (bR) proton transfer and their connection with the cation-binding role. We observe that both the efficiency of M formation and the kinetics of M rise and decay depend on the lipids and lattice but, as the lipids are removed, the cation binding is a much less important factor for the proton pumping function. Upon 75% delipidation using 3-[(cholamidopropyl)dimethylammonio]-propanesulfonate (CHAPS), the M formation and decay kinetics are much slower than the native, and the efficiency of M formation is approximately 30%-40% that of the native. Upon monomerization of bR by Trition X-100, the efficiency of M recovers close to that of the native (depending on pH), M formation is approximately 10 times faster, and M decay kinetics are comparable to native at pH 7. The same results on the M intermediate are observed if deionized blue bR (deI bbR) is treated with these detergents (with or without pH buffers present), even though deionized blue bR containing all the lipids has no photocycle. This suggests that the cation(s) has a role in native bR that is different than in delipidated or monomerized bR, even so far as to suggest that the cation(s) becomes unimportant to the function as the lipids are removed.