Microbial biomass and activity in subsurface sediments from Vejen,Denmark

Subsurface sediment samples were collected from 4 to 31 m below landsurface in glacio-fluvial sediments from the Quaternary period. The samples were described in terms of pH, electrical conductivity, chloride concentration, organic matter content, and grain size distribution. Viable counts of bacteria varied from 0.5 to 1,203 x 103 colony forming units/g dry weight (gdw); total numbers of bacteria acridine orange direct counts (AODC) varied from 1.7 to 147 × 10⁷ cells/gdw; growth rates (incorporation of [³H]-thymidine) varied from 1.4 to 60.7 × 10⁴ cells/(gdw · day); and rate constants for mineralization of ¹⁴C-labelled compounds varied from 0.2 to 2.3 × 10^–3 ml/(dpm · day) for acetate, and from 0 to 2.0 × 10^–3 ml/(dpm · day) for phenol. Sediment texture influenced the total number of bacteria and potential for mineralization; with increasing content of clay and silt and decreasing content of sand, AODC increased and the mineralization rate declined. Intrinsic permeability calculated from grain size correlated positively with mineralization rate for acetate. Statistical correlation analysis showed high correlations between some of the abiotic parameters, but it was not possible to point out a single abiotic parameter that could explain the variation of size and activity of the microbial population. The microbial data obtained in these geologically young sediments were compared to literature data from older sediments, and this comparison showed that age and type of geological formation might be important for the size and activity of the microbial populations.Offprint requests to: H.-J. Albrechtsen.     

Phenylalanine 4-monooxygenase from bovine and rat liver: Some physical and chemical properties

Phenylalanine 4-monooxygenase was purified from bovine liver using a modification of the procedure developed for the rat liver enzyme (Shiman, R., Gray, D. W., and Pater, A. 1979. J. Biol. Chem. 254:11300–11306). The enzyme preparation appeared essentially homogeneous on polyacrylamide gel electrophoresis under non-denaturing conditions. Electrophoresis in the presence of dodecyl sulfate revealed that about 95% of the protein had a mobility, corresponding to M_r=51,000. The remaining 5% was recovered in two minor bands corresponding to M_r of about 35,000 and 15,000 and is likely to result from limited proteolysis of the native enzyme with dissociation of the fragments on denaturation by detergent. The enzyme comigrated with the rat liver enzyme on polyacrylamide gel electrophoresis in both systems studied. No significant difference was observed between the amino acid composition of the bovine and rat liver enzyme, in the reactivity of their sulfhydryl groups or in their iron content (i.e. 1.5–3.0 iron atoms per peptide chain of M_r=50,000). Both enzymes contained less than 0.01 copper atom per peptide chain. The enzymes were inhibited in a similar manner by the chelator bathophenanthroline disulfonate (selective for iron and copper), but not by bathocuproine disulfonate (specific for copper). The results indicate that the bovine and rat liver enzymes are closely similar and that iron, but not copper, is essential for enzyme activity. High performance size-exclusion liquid chromatography revealed that both native enzymes exist in different oligomeric forms, but further studies are required to understand the physicochemical basis for this phenomenon.Abbreviations Bathophenanthroline 4,7-diphenyl-1, 10-phenanthroline - bathocuproine 2,9-dimethyl-4,7-diphenyl-1, 10-phenanthroline - Hepes N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - dansyl 1-dimethylaminonaphthalene-5-sulfonyl - DMPH₄ 2-amino-4-hydroxy-6,7-dimethyl-5,6,7,8-tetrahydropteridine - M_r relative molecular mass     

INCREASE IN SIZE AND CELL NUMBER OF LATERAL ROOT PRIMORDIA IN THE PRIMARY OF INTACT PLANTS AND IN EXCISED ROOTS OF PISUM SATIVUM AND VICIA FABA

Increase in cell number, and in anlage volume and length have been investigated during the development of lateral root primordia in roots of intact plants of Pisum sativum and Vicia faba and in excised roots of both species cultured in White's medium supplemented with 2% sucrose. With the exception of primordia in excised roots of Vicia, the general equation which best described increase in each aspect of primoridium growth measured against time was that for exponential growth. When the times necessary for cell number and primordium volume and length to double were determined at intervals over the period of development studied, however, they were found to vary. Similarly, estimates of the size of the proliferative fraction of cells at different times during anlage development indicated that this index of meristematic activity also fluctuated over the developmental period investigated, i.e., increase in cell number and in primordium volume and length do not occur in a truly exponential fashion as the primordia increase in size and cell number. One difference between anlage development in the roots of intact plants and in those grown in culture was that whereas the former primordia completed their development and emerged as lateral roots over the period of the investigation, the latter did not. Moreover, cell doubling time and anlage volume and length doubling times were longer, and the proliferation fraction of cells lower, over the whole period of, and at intervals during, primordium development in the excised roots compared with the results obtained for the roots of the corresponding intact plants.     

Increased Noradrenergic Metabolism in the Cerebellum of the Mouse Mutant Dystonia Musculorum

Abstract: The neurological mouse mutant dystonia musculorum exhibits bizarre appendicular and truncal dystonia without known cerebellar histopathology. We evaluated striatal dopamine and cerebellar norepinephrine metabolism in this mutant and compared the results with those obtained in wild-type BALB/c and B6C3 controls. Tyrosine hydroxylase activity and dopamine metabolite levels (homovanillic acid and 3,4-dihydroxyphenylacetic acid) in the striatum of the mutant were similar to controls. Tyrosine hydroxylase activity and the steady-state level of 3-methoxy-4-hydroxyphenethyleneglycol, a metabolite of norepinephrine, in the cerebellum were 38% and 42-66%, respectively, greater in the mutant. However, the level of norepinephrine was similar (∼350 ng/g). Further, a Purkinje cell-specific marker, cGMP-dependent protein kinase, was unchanged in the mutant and no Purkinje cell pathology was observed with light microscopy. The lack of Purkinje cell derangement and similar levels of cerebellar norepinephrine and cGMP-dependent protein kinase activity suggest that increased norepinephrine metabolism in the cerebellum of this mutant is not a morphological response to gross target cell loss during morphogenesis. The observed changes may be a reaction to abnormal impulse traffic or altered input/output pathways to the mutant cerebellum during its development.     

Specific changes in the production of prostanoids by the ovine cervix at parturition

A method of tissue superfusion has been used to measure prostanoid production by the ovine cervix during late pregnancy and at parturition. In late pregnancy (105–135 days of gestation) cervical tissue produced relatively large amounts of prostaglandin E (PGE); in comparison, the production rates of prostaglandin F (PGF), 13, 14-dihydro-15-oxo-prostaglandin F (PGFM) and 6-oxo-prostaglandin F_1α were generally low. Thromboxane B₂ (TXB₂) production was minimal and often unmeasurable. There were significant increases in the production rates of PGE and 6-oxo-PGF_1α by cervical tissue taken immediately after delivery, when compared to late pregnancy. Mean production rates of PGE increased from 19.8 ± 4.1 to 43.8 ± 7.4 ng/g. dry wt./min; 6-oxo-PGF_1α production rates increased more than three-fold from 10.0 ± 2.7 to 34.6 ± 9.8 ng/g. dry wt./min (means ± S.E.M.). There were no significant differences in the rates of production of PGF, PGFM and TXB₂ by the two groups.     

Analysis of development and growth in a mutant of Physarum polycephalum with defective cytokinesis

Cultures of amoebae of the mutant strain ATS23 isolated from strain CLd of Physarum polycephalum contain multinucleate cells and cells with increased nuclear DNA content. Plasmodia derived from ATS23 clones show abnormal morphology and defective sporulation. All abnormalities are enhanced by high incubation temperature (31 °C). Genetic analysis suggested that all the abnormalities were caused by a single mutation, denoted hts-23. The kinetics of plasmodium formation were followed in cultures of apogamic amoebae carrying hts-23 and hts⁺ (wild type) respectively. Results indicated that, relative to wild type, hts-23 did not increase the rate of plasmodium formation. There was evidence that, in both mutant and wild-type strains, commitment to plasmodium development occurred in uninucleate cells. Analysis of cell pedigrees by time-lapse cinematography indicated that the primary abnormal event in cultures of hts-23 amoebae was failure of cytokinesis; an apparently complete cleavage furrow was formed but cell separation failed, resulting in a binucleate cell. This event occurred randomly in pedigrees in which the majority of divisions were completed normally; its frequency increased during incubation at 31 °C. All other abnormalities in hts-23 amoebal cultures could be attributed to this primary event, assuming that DNA synthesis continued in the absence of cytokinesis and that the binucleate cells underwent the amoebal type of “open” mitosis, allowing the possibility of spindle fusion. This implies that the acquisition of “closed” mitosis is an essential early step in plasmodium development.     

Auditory sensitivity of the saccule in the American toad (Bufo americanus)

Summary Single unit recordings in the posterior nerve branchlet from the saccule have shown that, in the American toad (Bufo americanus), approximately 30% of the fibers respond to airborne sounds in a way similar to fibers from the two known auditory organs, the amphibian and basilar papillae. In response to tones, saccule fibers have best excitatory frequencies which fall into two disjoint populations: units in the low-frequency-sensitive group (below 300 Hz) show tone-on-tone suppression while those in the high-frequency-sensitive group (700–1,200 Hz) show no evidence of peripheral inhibition. Saccule units have somewhat higher thresholds than those from the other auditory organs. It is suggested that the high-frequency-sensitive fibers might be useful for discriminating mating calls in an intense chorus while the low-frequency-sensitive units likely respond to other high intensity sounds in the environment.Research supported by the U.S. Public Health Service (NIH grant NS-09244).