Redundant transfer of F' plasmids occurs between Escherichia coli cells during nonlethal selections.

Surface exclusion is the mechanism by which F plasmids prevent the redundant entry of additional F plasmids into the host cell during exponential growth. This mechanism is relaxed in cells that are in stationary phase. Using genetically marked F' plasmids and host strains, we extend this finding to Escherichia coli populations during extended nonlethal selection in bacterial lawns. We show that a high level of redundant transfer occurs between these nongrowing cells during the selection. This result has implications for the mechanism of adaptive mutagenesis.     

Uptake and Release of d-Aspartate, GABA, and Glycine in Guinea Pig Brainstem Auditory Nuclei

Abstract: This study attempts to determine if the medial (MSO) and lateral superior olive (LSO), medial nucleus of the trapezoid body (MNTB), ventral nucleus of the lateral lemniscus (VNLL), and central nucleus of the inferior colliculus (ICc) contain glutamatergic synaptic endings. Micropunch and microdissection procedures provided fresh samples of these auditory nuclei for the measurement of the high-affinity uptake and electrically evoked release of exogenous d -[³H]ASP. The study also determined if the LSO and MSO contain glycinergic synaptic endings by measuring uptake and release of [¹⁴C]-Gly in these nuclei, and whether the MNTB, VNLL, and ICc contain GABAergic endings by assessing the uptake and release of [¹⁴C]GABA in these structures. Several strategies optimized the evoked Ca²⁺-dependent release of the labeled amino acids. These included the enhancement of high-affinity uptake during loading of the markers into the tissues, inhibition of uptake during the subsequent measurement of release, and use of an electrical stimulus current that evoked maximal Ca²⁺-dependent release. Each of these nuclei manifested the high-affinity uptake and the evoked Ca²⁺-dependent release of d -[³H]Asp, suggesting the presence of synaptic endings that may use Glu or Asp as a transmitter. Similar findings suggest the presence of glycinergic synaptic endings in the LSO and MSO, and of GABAergic synaptic endings in the MNTB, VNLL, and ICc.     

Evidence for Glutamatergic Projections from the Cochlear Nucleus to the Superior Olive and the Ventral Nucleus of the Lateral Lemniscus

Abstract: This study attempts to determine if projections ascending from the guinea pig cochlear nucleus (CN) could be glutamatergic and/or aspartatergic. Multiple radio frequency lesions were made to ablate the right CN. The ablation was verified histologically. To identify the principal targets of CN efferents, silver impregnation methods were used to localize the preterminal degeneration of fibers in transverse sections of the brainstem 5 and 7 days after CN ablation. CN efferents projected heavily to the lateral superior olive (LSO) ipsilaterally, the medial superior olive (MSO) bilaterally, and contralaterally to the medial (MNTB) and ventral (VNTB) nuclei of the trapezoid body, the ventral (VNLL) and intermediate nuclei of the lateral lemniscus and the central nucleus of the inferior colliculus (ICc). There were smaller projections to the lateral nucleus of the trapezoid body ipsilaterally, the dorsal and dorsomedial periolivary nuclei bilaterally, and the dorsal nucleus of the lateral lemniscus contralaterally. There were sparse projections to the VNLL and ICc ipsilaterally and the CN contralaterally, and a very sparse projection to the contralateral LSO. To determine if CN efferents were glutamatergic and/or aspartatergic, the fresh brainstem was sectioned transversely and samples of the LSO, MSO, MNTB, VNLL, and ICc were taken to measure the electrically evoked release and the uptake of d -[³H]Asp and [¹⁴C]Gly or [¹⁴C]GABA 3–5 days after the CN ablation. The release studies suggest that only certain of the histologically identified projections ascending from the CN may be glutamatergic and/or aspartatergic. CN ablation depressed d -[³H]Asp release in the MSO bilaterally and in the contralateral MNTB and VNLL, suggesting that the CN efferents to these nuclei may use glutamate or aspartate as a transmitter. It was unclear whether a marginal depression of d -[³H]Asp release in the ipsilateral LSO reflected the presence of glutamatergic CN projections to this nucleus. d -[³H]Asp release in the ICc was unaffected, suggesting that CN efferents to this nucleus may not be glutamatergic. There were no deficits in d -[³H]Asp uptake. [¹⁴C]Gly release from the LSO and MSO was unchanged. [¹⁴C]Gly uptake was unchanged in the MSO and depressed only in the contralateral LSO, possibly reflecting subnormal uptake activity in endings contributed by contralateral MNTB cells that had lost their CN efferents. [¹⁴C]GABA uptake in the MNTB, VNLL, and ICc was unchanged. [¹⁴C]GABA release was unchanged in the VNLL and ICc. [¹⁴C]GABA release was depressed only in the contralateral MNTB, possibly reflecting the loss of a small complement of GABAergic CN efferents and the reaction of GABAergic projections from the contralateral VNTB to their loss of CN efferents.     

Identification of Frankia Strains by Two-Dimensional Polyacrylamide Gel Electrophoresis

Fifteen Frankia strains from five different plant species were analyzed by two-dimensional polyacryl-amide gel electrophoresis to determine their relatedness by comparing the polypeptide patterns obtained. Three major subgroups (A, C, and D) were found in the Alnus-Comptonia-Myrica cross-inoculation group. An isolate from Purshia tridentata had a unique protein pattern and represents a distinct group of frankiae. Members of group A were isolated from root nodules of Alnus incana subsp. rugosa and Alnus viridis subsp. crispa. Group C organisms were from A. incana subsp. rugosa and Comptonia peregrina nodules, and group D organisms were from A. incana subsp. rugosa, A. viridis subsp. cripsa, and Myrica pensylvanica root nodules. Isolates from each gel group were obtained at several widely separated geographical locations. The results indicate that two-dimensional polyacrylamide gel electrophoresis is useful for identifying Frankia isolates.     

Japanese Hollies: Intolerant Hosts of Meloidogyne arenaria in Microplots

Japanese hollies were itttolerant of Meloidogyne arenaria in field microplot experiments. Ilex crenata var. rotundifolia was relatively more tolerant than I. crenata var. convexa or I. crenata var. helleri. When M. arenaria was added at various itfitial population densities to soil containing plants of "Helleri," "Convexa," and "Rotundifolia," respectively, 91, 75, and 25% were killed by the end of the third growing season. No control plants died during the same period. Initial numbers of M. arenaria larvae and eggs were the only population densities that were correlated (negatively), regardless of cultivar, with plant growth over the three growing seasons. A linear relation was found for initial density of M. arenaria and growth of I. crenata rotundifolia. Increasing nematode density by 10-fold suppressed the growth of this cultivar by 23%.     

Fractionation of primary amyloid fibrils. Characterization and chemical interaction of the subunits.

Amyloid fibrils of kappa origin from a patient with primary amyloidosis are dissociated in various denaturants and fractionated into their subunit components on Sepharose 6B. Solubilization of the fibrils in 4 M guanidine-HCl followed by reduction and alkylation produced 22 000 and 17 000 dalton fractions. Without prior reduction and alkylation, these fractions exist as a high molecular weight protein which can be separated on Sepharose 6B. A high molecular weight protein can be directly dissociated from the amyloid fibril with 1% sodium dodecyl sulfate or 1 M NaCl. Reduction and alkylation of this material produces the two lower molecular weight fractions, i.e., 22 000 and 17 000. These have in the first 20 residues identical N-terminal amino acid sequences; they share immunologic identity and have similar tryptic peptide map profiles. Amino acid analysis of the 22 000 dalton fraction is identical with the intact immunoglobulin light chain isolated from the patient's serum. These data suggest that the insoluble amyloid fibril is the result of aggregation by disulfide linkages between the 22 000 and 17 000 dalton fractions.     

13C NMR spectroscopy of Pyrrolizidine alkaloids

The ¹³C NMR spectra of nine pyrrolizidine alkaloids of the macrocyclic diester type, seven of the corresponding N-oxides and of the parent base retronecine have been recorded and the signals assigned. The 13C NMR signals were found to be sensitive to structural variation in both the diester moiety and the heterocyclic ring system, providing useful information for structural elucidation, particularly when the ¹H NMR spectra may be difficult to interpret.