The influence of fluid mixing upon respiratory patterns for extended growth of a methylotroph in an air-lift fermenter

The influence of methanol dispersion and fluid mixing upon respiratory patterns observed during unlimited fedbatch growth of the methylotrophic bacterium Methylophilus methylotrophus has been investigated. A concentric tube air-lift fermenter was employed for which the mixing and fluid circulation patterns have been well characterized. Respiratory quotients showed a marked dependence upon the position in the vessel at which methanol was injected, the volumetric rate of such methanol addition, the fluid circulation time, and the local mixing behavior; the latter two factors of which are both determined by the air throughput. Such variations are discussed on the basis of simple mixing concepts and observations of fluid dispersion.     

Light control of extractable nitrate reductase activity in higher plants

Light regulation of extractable nitrate reductase (NR) activity of higher plants is complicated by: 1) involvement of several photoreceptors, 2) differences in the relative importance of the several photoreceptors among species and among developmental stages of the same species, 3) two types of effects – alteration of activity of existing NR and influences on de novo synthesis of NR, and 4) differing forms of NR within the same species. The interrelationships of all of these factors are not clear. It may be that each system will have to be understood separately before a general model can be developed. Immunochemical quantification of NR from systems exposed to varied light regimes may enhance our understanding of this area. Currently few general conclusions can be made; however, we think that the following statements are true or are usually true: (1) Phytochrome influences extractable NR activity by the low irradiance response and high irradiance response in etiolated tissues. (2) In de-etiolated tissues phytochrome can influence NR activity decay at the end of a light period by the low irradiance response. (3) The phytochrome equilibrium or the absolute level of P_fr influences extractable NR activity in green tissues under white light. (4) Blue light influences extractable NR activity through phytochrome and another, unknown, blue light-absorbing pigment. Flavins may be involved in vitro in reactivation of inactivated NR. (5) Photosynthesis does not directly influence the induction of the forms of NR that require substrate and light for induction. (6) In some tissues there appears to be a close link between nitrite-reducing and nitrate-reducing capabilities. (7) Much circumstantial evidence from kinetic and protein-synthesis-inhibitor studies and the only available immunochemical data indicate that light induces de novo synthesis of NR, resulting in increased extractable activity.     

Transposon mutagenesis of Rhizobium japonicum

Summary We report here successful mutagenesis with Transposon Tn5 of three slow-growing strains of Rhizobium japonicum USDA 122, 61A76, USDA 74 and one fast-growing strain, USDA 191. Strains were chosen as representatives of different DNA homology and serogroups of this divergent species, which effectively nodulate North American soybean cultivars. The source of Tn5 was the suicide plasmid pGS9, which possesses broad host range N-type transfer genes in a narrow host range p15A replicon. The selection of Tn5 mutants was facilitated by the expression of the Tn5 encoded streptomycin gene in R. japonicum. Kanamycin and streptomycin resistant colonies appeared from interspecific crosses with E. coli at optimal frequencies of 10^-6 for R. japonicum USDA 61A76 and USDA 191 and 5x10^-7 for R. japonicum USDA 122 and USDA 74. Altogether, 6550 Tn5 mutants were isolated in USDA 122 and 61A76, and a small number from USDA 74 and USDA 191. Colony hybridization showed that all tested mutants of 61A76 and USDA 122 contained Tn5. Physical analysis of total DNAs from representative numbers of USDA 122, 61A76 and USDA 191 mutants revealed that each of them carried one copy of the transposon integrated randomly in the genome. This was also true for most USDA 74 mutants. Screening of mutants for auxotrophy showed frequencies of 0.2% for USDA 122 and 0.08% for 61A76. Several symbiotically defective mutants were identified on plants, Glycine soja and G. max.     

The nucleotide sequence of an infectious clone of the geminivirus beet curly top virus

A number of infectious clones of a Californian isolate of the leafhopper-transmitted geminivirus beet curly top virus (BCTV) have been constructed from virus-specific double-stranded DNA isolated from infected Beta vulgaris and used to demonstrate a single component genome. The nucleotide sequence of one infectious clone has been determined (2993 nucleotides). Comparison with other geminiviruses has shown that the organisation of the genome closely resembles DNA 1 of the whitefly-transmitted members. The four conserved coding regions of DNA 1 have highly homologous counterparts in BCTV with the exception of the putative coat protein which is more closely related to those of the leafhopper-transmitted geminiviruses suggesting a strong interrelationship between coat protein and insect vector. A BCTV component equivalent to DNA 2 is not required for virus infection or transmission and has not been isolated from infected plants.     

Nitrospira marina gen. nov. sp. nov.: a chemolithotrophic nitrite-oxidizing bacterium

A new chemolithotrophic nitrite-oxidizing bacterium, for which the name Nitrospira marina is proposed, was isolated from the Gulf of Maine. N. marina is a Gramnegative curved rod which may form spirals with 1 to 12 turns. Cells have a unique periplasmic space and lack intracytoplasmic membranes and carboxysomes. N. marina is an obligate chemolithotroph, but best growth is obtained in a mixotrophic medium. N. marina may be one of the most prevalent nitrite-oxidizing bacteria in some oceanic environments. Type strain is field with American Type Culture Collection (ATCC 43039).     

Distribution of intrapleural and intravenous Corynebacterium parvum in humans; 99mTc−, and 131I-labeled bacteria

Summary The distribution of Corynebacterium parvum labeled with ¹³¹iodine or ^99mtechnetium was studied in 17 patients with bronchogenic carcinoma. The labeled bacteria were given intravenously or intrapleurally and monitored by whole-body gamma tracking and samples of blood and urine. Even though the rate of physical decay is quite different for ¹³¹iodine and ^99mtechnetium, the tracking time of labeled bacteria was limited to 24 h after injection for both radioactive isotopes. Technetium labeling was preferred because of greater imaging resolution and less radiation dose to the patient. Following intravenous administration, labeled C. parvum was found predominantly in the liver and spleen, and in a lesser amount in the lung. Radioactivity was confined to the pleural cavity after intrapleural injection. These results suggest the combined intravenous and intrapleural route of adjuvant immunosupportive agents such as C. parvum for operable lung cancer patients.     

Preparation of a cell-free extract from rat brain which can initiate protein synthesis in vitro

A cell-free protein synthesis system, derived from brains of 3 mo-old male Fischer-344 rats, has been characterized. The optimum conditions for amino acid incorporation in the system were 5 mM magnesium ion and 200 mM potassium ion. Incorporation depended on the addition of ATP, GTP, and an enegy-generating system, and was sensitive to addition of the drugs aurintricarboxylic acid and sodium fluoride, inhibitors of initiation of protein synthesis. Both 40S and 80S initiation complexes were labeled in vitro, using [³⁵S]methionine. Such labeling was sensitive to the protein synthesis inhibitors, aurintricarboxylic acid and sodium fluoride. The system, which can initiate protein synthesis, should be of use for examining mechanisms which underlie alterations in rat brain protein synthesis induced by various treatments.     

Solubilization and assay of a colony-stimulating factor receptor from murine macrophages

The colony-stimulating factor, CSF-1, selectively stimulates the survival, proliferation, and differentiation of mononuclear phagocytes. The solubilization, assay, and characteristics of the CSF-1 receptor from the J774.2 murine macrophage cell line are described. The recovery of cell-surface receptor in the postnuclear supernatant membrane fraction of hypotonically disrupted cells was 76%. Recovery of the ligand binding activity of the receptor after solubilization of this fraction with 1% Triton X-100 was approximately 150%. The binding of 125I-CSF-1 to intact cells and membrane preparations was consistent with the existence of a single class of high-affinity receptor sites. In contrast, the equilibrium binding of 125I-CSF-1 to the solubilized postnuclear fraction indicated the existence of two distinct classes of binding site (apparent Kds 0.15 nM and 10 nM). A rapid assay was developed for the high-affinity sites, which were shown to be associated with the CSF-1 receptor. The function of the low-affinity sites, which have not been demonstrated on intact cells or cell membranes and which are 13 times more abundant than the high-affinity sites, is unknown. The solubilized high-affinity receptor-CSF-1 complex was stable on storage at 0 degrees C and -70 degrees C but dissociated at 37 degrees C. Dissociation also occurred at 0 degrees C in buffers of low pH (4.0) or high ionic strength (0.7 M NaCl).