Host-vector systems for the thermotolerant methanol-utilizing bacteriaMethylophilus spp. KISRI-Strains

Fourteen recombinant plasmids were constructed by inserting fragments of pSAS, a naturally occurring plasmid ofMethylophilus spp. KISRI-5, into the multiple cloning sites of pUC19. Six recombinants and three knownEscherichia coli plasmids were used to transform three thermotolerant methylotrophic KISRI strains by use of an optimized protocol of electroporation. Analysis of transformants for plasmid DNA showed that all plasmids were stable in the methylotrophic hosts. These studies offer opportunities to developMethylophilus spp. as host-vector systems.     

Morphological variation of Echinococcus granulosus protoscoleces from hydatid cysts of human and various domestic animals in Jordan

, and 1988. Morphological variation of Echinococcus granulosus protoscoleces from hydatid cysts of human and various domestic animals in Jordan. International Journal for Parasitology 18: 1111–1114. Rostellar hook morphology of protoscoleces was employed to study the possible existence of Echinococcus granulosus strains in humans and various domestic animals in Jordan. A distinct form in the donkey was evident as the protoscoleces from this host did not share any characteristics with those from the other hosts examined. Sheep, goats and cattle appeared to be affected by another form since the protoscoleces from their hydatid cysts shared six out of nine characteristics studied. Protoscoleces of camel and human cysts shared seven out of nine characteristics studied and they were different in six characteristics from protoscoleces from other hosts. Differences observed among the three forms may reflect strain variation of E. granulosus in this country.     

Absence of prostaglandins in semen of men with cystic fibrosis is an indication of the contribution of the seminal vesicles

Five men with cystic fibrosis delivered semen samples which were analysed for the content of the four main groups of prostaglandins (PGs). No PGs could be found in any of the specimens. The results of the present study provide further evidence that the seminal vesicles constitute the dominant site of production of PGs in human semen.     

Alterations in the number of rRNA operons within the Bacillus subtilis genome

Deletions and additions of rRNA gene sets in Bacillus subtilis were observed by Southern hybridizations using cloned radiolabeled rDNA sequences. Of the ten rRNA gene sets found in B. subtilis 168M or NCTC3610, one was deleted in strains possessing the leuB1, ilvC1, argA2 and pheA1 mutations. Among EcoRI restriction fragments of genomic DNA products, a 2.9-kb 23S rRNA homolog was missing. In HindIII digest, both 5.5- and 5.1-kb hybrid bands were lost with 16S and 23S probes, respectively. Similarly, genomic DNAs digested with SmaI showed the absence of both 2.1- and 2.0-kb fragments that hybridized to 16S and 5S sequences, respectively, in wild-type genomes. In contrast, B. subtilis strain 166 and its derivatives displayed a gain of a 3.3-kb HindIII fragment homologous to 16S rRNA. Transforming the ilvC1 and leuB1 mutations into new genetic backgrounds revealed in some clones the concomitant introduction of the ribosomal defect. Transformations with the slightly heterologous donor DNA from strain W23 yielded some Leu+ and Arg+ transformants with altered hybridization patterns when probed with cloned sequences. We propose that the deletion of the rRNA operon occurred in the ilv-leu gene cluster of the B. subtilis genome as a result of unequal recombination between redundant sequences.     

Neolignans from Ocotea catharinensis

The trunk bark of Ocotea catharinensis yielded, besides the known bicyclo(3.2.1)octanoid neolignans canellin-C and 5′-methoxycanellin-C, two epimers rel-(1R,4S and 4R,5S,6R,7S,8R)-1-allyl-4,8-dihydroxy-3,5-dimethoxy-7-methyl-6-piperonyl-bicyclo(3.2.1)oct-2-enes and rel-(1R,5S,6R,7S,8R)-1-allyl-3,8-dihydroxy-5-methoxy-7-methyl-6-piperonyl-4-oxobicyclo(3.2.1)oct-2-ene. The hydrobenzofuranoid neolignans are represented by the equally novel (2S,3S,5R)-5-allyl-5,7-dimethoxy-3-methyl-2-piperonyl-2,3,5,6-tetrahydro-6-oxobenzofuran and (2R,3S,3aS)-3a-allyl-5,7-dimethoxy-3-methyl-2-piperonyl-2,3,3a,6-tetrahydro-6-oxobenzofuran.     

Macrophage regulation of DNA synthesis in lymphoid cells: effects of a soluble factor from macrophages

Addition of rat peritoneal macrophages to nonadherent rat spleen cells in culture results in enhancement or suppression of DNA synthesis depending on the ratio of macrophages to lymphocytes. At high ratios of macrophages to lymphocytes (1:5), suppression can be observed as early as four hours. Macrophages suppress incorporation of thymidine (TdR) by nonadherent spleen, thymus and bone marrow cells, in most instances, to less than 5% of that observed in culture to which macrophages were not added. In the presence of macrophages, incorporation of [³H]uridine and [¹⁴C] amino acids by spleen cells was also moderately suppressed. Based on ⁵¹Chromium release and dye exclusion assays, it appears that suppression is not due to cytotoxicity. Furthermore, suppression of [³H]TdR incorporation by nonadherent spleen cells is reversible, in the presence of an antigenic stimulus, following removal of the macrophages from the cultures. The suppressive effects are not elicited by extracts of macrophages, freeze-thawed or heated macrophages, but appear to be due to a low molecular weight, heat stable factor released into the macrophage culture fluid.     

Sterols in species of Pythium

Summary Analysis by gas chromatography revealed the presence of small amounts of squalene, but not lanosterol nor ergosterol in Pythium paroecandrum, P. ultimum, P. graminicola, and P. arrhenomonas. However, when acetate-¹⁴C was used as a precursor for sterols, even squalene was not found in P. graminicola. The deficiency in the sterol synthesizing mechanism may therefore be at or before the squalene forming step. Both squalene and ergosterol were present in the mycelium of Rhizoctonia solani, as shown by both gas chromatography and by the incorporation of acetate-¹⁴C into ergosterol. The absence of ergosterol in Pythium and its presence in Rhizoctonia is consistant with the resistance to the antibiotic filipin of Pythium species and the sensitivity of R. solani.     

The effect of sterols on the metabolism of Pythium species

Summary The growth of several Pythium species is increased between 65 and 100% if cholesterol is added to the growth medium. The optimum concentration is 15 mcg per ml. Mycelium of Pythium ultimum, in which cholesterol is present, incorporates glucose-U-¹⁴C and releases ¹⁴CO₂ at a faster rate than the corresponding sterol free mycelium. In sterol containing cells, more ¹⁴CO₂ is produced from a given amount of absorbed glucose-U-¹⁴C than in sterol free cells, there is thus in sterol containing hyphae a higher level of energy production. This condition can account for the increase in growth due to cholesterol. Only if sterols are present in the cellular membranes of Pythium species is the optimum synthetic capacity reached.     

Changes in fungi with age. II. Respiration and respiratory enzymes of Rhizoctonia solani and Sclerotium bataticola

Van Etten, James L. (University of Illinois, Urbana), H. Peter Molitoris, and David Gottlieb. Changes in fungi with age. II. Respiration and respiratory enzymes of Rhizoctonia solani and Sclerotium bataticola. J. Bacteriol. 91:169-175. 1966.-The rate of respiration of Rhizoctonia solani and Sclerotium bataticola decreased with age. This decrease in respiratory rate might be produced by a decrease in the specific activity of one or more enzymes involved in carbohydrate metabolism. Specific activities in cell-free extracts were measured for most of the enzymes in the hexose monophosphate shunt, Embden-Meyerhof-Parnas pathway, tricarboxylic acid cycle, and terminal electron-transport system. In addition, glucose oxidase, isocitritase, and malic enzyme were measured. In R. solani, increases in activity with age occurred for hexokinase, alpha-glycerolphosphate dehydrogenase, malic dehydrogenase, and cytochrome oxidase. Decreases occurred for phosphohexokinase, aconitase, nicotinamide adenine dinucleotide-specific isocitric dehydrogenase, reduced nicotinamide adenine dinucleotide oxidase, and at least one of the enzymes between 3-phosphoglycerate and pyruvate. In S. bataticola, increases in activity with age were observed for phosphohexokinase, pyruvic dehydrogenase, fumarase, malic dehydrogenase, and malic enzyme, whereas none of the enzymes decreased. The specific activities of the remaining enzymes did not change with age in either fungus.