Versatile shuttle cloning vectors for the unicellular cyanobacterium Anacystis nidulans R2

Abstract 3 new shuttle cloning vectors for gene transfer into Escherichia coli and Anacystis nidulans have been constructed by utilizing the cyanobacterial origin of replication of the small plasmid pANS from A. nidulans . 2 of these new vectors, pXB7 (pDPL13 derivative) and pECAN8 (pUC8 derivative), convey ampicillin resistance, and transform A. nidulans with relatively high frequencies. Vector pXB7 has 10 unique cloning sites; pECAN8 contains 4 cloning sites within the lacZ gene permitting rapid detection of DNA inserts in the presence of Xgal. The third vector, pKBX, has a lower transformation frequency but adds kanamycin resistance as a selectable gene for shuttle vectors of cyanobacteria.     

Late enzymes of vindoline biosynthesis

From differentiated plants of Catharanthus roseus (L.) G. Don we have isolated a specific enzyme of the vindoline biosynthetic pathway catalysing the S-adenosylmethionine-dependent methylation of 11-O-demethyl-17-O-deacetyl-vindoline. The enzyme we named S-adenosyl-L-methionine : 11-O-demethyl-17-O-deacetylvindoline 11-O-methyltransferase. This transferase exhibits a high substrate specificity. Obviously the O-methylation at C-11 precedes the O-acetylation at the C-17 position during the biosynthesis of vindoline.A second enzyme was detected which hydrolyses the acetyl function of vindoline. The distribution of this acetylesterase in C. roseus plants demonstrates that the enzyme is not specifically associated with the vindoline distribution in the plant material. Most probably this enzyme plays no essential role in the biosynthesis of vindoline.     

Comparative analysis of caffeine and 3-aminobenzamide as DNA repair inhibitors in Syrian baby hamster kidney cells

The effects of caffeine and 3-aminobenzamide (3-AB) on Syrian baby hamster kidney cells treated with DNA-alkylating agents and ultraviolet-light suggest that two different DNA-repair mechanisms are involved. Both these agents enhanced the cell kill after methyl methanesulfonate (MMS) treatment. However, enhanced lethality was observed only with caffeine post-treatment when cells were exposed to nitrogen mustard (HN2) or ultraviolet light (UV); 3-AB did not appreciably change cell killing by these agents. With MMS-treated cultures, the effect of caffeine was maximal about 16 h later. The effect of 3-AB on the other hand, was exerted during the first 4 h after exposure to MMS. Caffeine's effect on cell survival could be abolished by low concentrations of cycloheximide, whereas 3-AB's effect could not. Furthermore, the G2 block in cell cycle progression, after MMS treatment, was not observed if the cells were post-treated with caffeine. In the presence of 3-AB, MMS-treated cells were arrested in G2 phase at a much earlier time compared to cells not treated with 3-AB. Finally caffeine post-treatment produced a 10-fold increase in nuclear fragmentation in MMS-treated cells. 3-AB did not cause nuclear fragmentation by itself but further enhanced the nuclear fragmenting effect of caffeine when both agents were present during the posttreatment. Therefore, we propose that 3-AB and caffeine each prevent a different repair mechanism from being effective.     

Inhibition of in vitro germination and spherulation ofCoccidioides immitis byAllium sativum

An aqueous extract of a dehydrated garlic preparation with uniform consistency inhibited all eight clinical isolates of the dimorphic fungus,Coccidioides immitis. The inhibitory and lethic concentrations were in the range of 3.12–6.25 mg/ml for both the saprophytic (mold) and parasitic (spherule) forms ofC. immitis. At 6.25-mg/ml concentration, the organism lost its viability within 6 h. The conversion of arthroconidia into spherules in a chemically defined liquid medium was prevented by garlic extract diluted to 1:320 (3.12 mg/ml). The data indicate that components of garlic readily inhibited the in vitro germination and spherulation of this medically important dimorphic fungus.     

Leukocytic Hypersensitivity in Experimental Group A Streptococcal Infections

Skin hypersensitivity to streptococcal antigens was demonstrated in 15 New Zealand red rabbits that were sensitized by intracutaneous injections with group A streptococci, and leukocytic hypersensitivity, as determined by in vitro inhibition of leukocyte migration, was demonstrated in 11 of the rabbits. The skin hypersensitivity persisted for at least 8 weeks, whereas the leukocytic hypersensitivity generally waned rather rapidly. The leukocytic hypersensitivity reappeared in the infected rabbits that had developed this hypersensitivity. However, it did not reappear on reimmunization with living streptococci of the type originally employed, whereas it did reappear with living but not heat-killed streptococci of another M type.     

Preparation of anti-inhibitor serum and its effect on the multiplication of influenza virus

Antiserum был подготовлен противингибитор вируса haemagglutination (IVH)одна из эмбриона chorioallantoic мембр анныепутем иммун изации морских св инокЭто анти-инги битора в сыворотке заблокирован ингибирование IVH из chorioallantoicмембран в haemagglutination испытанияно ника кого влияния на ре цепторы эритроци товСыворотке кро] ви не влияет рост chorioallantoic мембраны клет ок инет нейтрализ овать воздействи е на вирус гриппа Она снизилась спо собность chorioallantoic мембр ан для adsorb вируса гри ппа, а также степен ь размножения вир усав такого рода тканиАвторы хоте ли бы поблагодари ть г-н Г. Ruttkay-Ne -палубе и для проведения эле ктрофореза меру ния.