Vertical nitrate fluxes in the oligotrophic ocean.

The vertical flux of nitrate across the thermocline in the upper ocean imposes a rigorous constraint on the rate of export of organic carbon from the surface layer of the sea. This export is the primary means by which the oceans can serve as a sink for atmospheric carbon dioxide. For the oligotrophic open ocean regions, which make up more than 75% of the world's ocean, the rate of export is currently uncertain by an order of magnitude.

Aztreonam concentrations in human tissues obtained during thoracic and gynecologic surgery.

The concentrations of aztreonam in human tissues obtained during surgery were measured after a single 2-g intravenous dose. The average concentration in the skeletal muscle, atrial appendage, lung, sternum, pericardial fluid, endometrium, myometrium, fallopian tube, and ovary varied from 3 to 33 micrograms/g (or microgram/ml). These concentrations significantly exceed the MIC for 90% of strains for most members of the family Enterobacteriaceae.

The rho-115 mutation in transcription termination factor rho affects its primary polynucleotide binding site.

We have investigated the effect of the rho-115 mutation on the catalytic properties of the Escherichia coli termination protein, rho. Comparison of the primary and secondary polynucleotide binding sites activities reveals dramatic differences between the mutant and wild-type molecules. Wild-type rho must bind single-stranded polynucleotides to activate its nucleotide triphosphatase (NTPase) activity, and either poly(C), or poly(dC) plus oligo(C), will suffice.

An examination of the prepatent period and the absence of a crowding effect in Angiostrongylus cantonensis in laboratory rats.

The effects of the size of the infective inoculum on the prepatent period and the effect of crowding on body volume were examined in populations of Angiostrongylus cantonensis (Metastrongyloidea: Angiostrongylidae) in the laboratory rat. Groups of five rats were infected with 5, 10, 20 and 40 L3 of A. cantonensis, respectively.

Maturation of Escherichia coli tryptophan operon mRNA: evidence for 3' exonucleolytic processing after rho-dependent termination.

The mature 3' end of Escherichia coli tryptophan operon mRNA in vivo coincides with a site (trp t) having features commonly associated with rho-independent terminators in bacteria. Efficient generation of this 3' end in vivo is nevertheless affected by a distal rho-dependent site (trP-t'), though these two sites behave independently in vitro. We have cloned these sites upstream of the galactokinase gene (galK), and galactokinase levels in vivo indicate that, as terminators per se, their efficiencies (37% for trp t, and 79% for trp t') do not differ significantly from those observed in vitro.

Penetration of aztreonam into human cerebrospinal fluid in the presence of meningeal inflammation.

Cerebrospinal fluid of aztreonam were measured in 11 patients with meningeal inflammation. Two to eight hours after a single 2 gm intravenous dose, CSF aztreonam levels ranged from 0.76 to 16.

Escherichia coli transcription termination factor rho has a two-domain structure in its activated form.

Limited tryptic digestion of Escherichia coli transcription termination factor rho [an RNA-dependent nucleoside triphosphatase (NTPase)] yields predominantly two fragments (f1 and f2) when the protein is bound to both poly(C) and ATP. The apparent molecular masses of the two fragments are 31 kDa for f1 and 15 kDa for f2, adding up to the molecular mass of the intact rho polypeptide chain (46 kDa). Sequence analysis of the amino termini demonstrates that f1 is derived from the amino-terminal portion of rho and that the trypsin cleavage that defines f2 occurs at lysine-283.

Pharmacokinetics of aztreonam in elderly male volunteers.

The pharmacokinetics of aztreonam were studied in 12 healthy male volunteers aged 65 to 75 years who received 1 g of the antibiotic intravenously. Data were fitted to a two-compartment open model to yield the following parameters: t 1/2, lambda 1, 0.15 h; t 1/2,z, 2.

Maximizing gene expression from plasmid vectors containing the lambda PL promoter: strategies for overproducing transcription termination factor rho.

We have constructed two plasmids in which transcription of the rho gene from Escherichia coli K-12 is under the control of the lambda phage PL promoter. In p31-356, the normal rho promoter is deleted, but the remainder of the rho leader region, including the ribosome binding site, is present. In p39-AS, the rho leader is completely absent, and the lambda cII ribosome binding site replaces that of rho.

Penetration of aztreonam into cerebrospinal fluid of patients with and without inflamed meninges.

Aztreonam was administered as a single, 2-g intravenous dose to 25 patients with noninflamed meninges and to 9 patients with inflamed meninges. It was well tolerated and was detected in the cerebrospinal fluid at the initial sampling period at 1 h after the end of infusion. Aztreonam levels in the cerebrospinal fluid of patients with inflamed meninges were four times higher than those recorded for the same time period in patients with noninflamed meninges.

Targeted mutagenesis in vitro: lac repressor mutations generated using AMV reverse transcriptase and dBrUTP.

We have cloned the gene for the lac operon repressor (lacI) of Escherichia coli into the M13 related phage f1. Mutagenesis of the lacI gene was performed in vitro by filling dsDNA molecules gapped over the lacI gene with Avian Myeloblastosis Virus (AMV) reverse transcriptase. LacI mutants are found at a frequency of 1 in 10(4) using a genetic screen in vivo.

Toxicity of 2-deoxygalactose to Saccharomyces cerevisiae cells constitutively synthesizing galactose-metabolizing enzymes.

Analysis of 400 independent spontaneous mutations conferring 2-deoxygalactose resistance upon cells constitutive for the galactose pathway suggests that toxicity is due to 2-deoxygalactose-1-phosphate. Selection for and against growth on galactose in the same strain is now possible; application to systems with transcriptional or translational gene fusions to galactokinase are discussed.

Pharmacokinetic interaction of aztreonam with other antibiotics.

The pharmacokinetic interaction of the monobactam antibiotic aztreonam with cephradine, clindamycin, gentamicin, metronidazole, and nafcillin was investigated in five separate studies in 48 healthy male volunteers. All drugs were administered by 30-minute intravenous infusions in single-dose, three-way balanced cross-over studies. Drug levels were measured in serum, protein-free filtrate of serum, and urine.

Rho-dependent termination and concomitant NTPase activity requires a specific, intact RNA region.

We have investigated the specific DNA and RNA requirements for rho-dependent transcription termination in vitro. As a model, we have used templates containing the rho-dependent terminator of the Escherichia coli trp operon, trp t'. Templates containing the trp t' region direct specific rho-dependent termination in vitro, with concomitant stimulation of the rho NTPase activity, and deletion of the trp t' region results in templates that do not induce rho-dependent termination or rho NTPase activity.

Nursing enhances the negative effect of estrogen on LH release in the cow.

Twenty-three crossbred beef cows between 4 and 5 yr of age were assigned at random to one of six treatments: (1) ovariectomized 4 d postpartum (OVX) with early weaning of calves 21 d postpartum (OVX-EW; n = 4), (2) OVX-EW and 17 beta-estradiol implants (OVX-E2-EW; n = 4), (3) OVX and normal nursing by calves throughout the experiment (OVX-NN; n = 3), (4) OVX-NN and 17 beta-estradiol implants (OVX-E2-NN; n = 4), (5) intact cows and early weaning of calves 21 d postpartum (EW), (6) intact cows and normal nursed (NN). Blood was collected at 15-min intervals over a 4-h period once weekly during the 12-wk postpartum period in the OVX cows. Early weaned intact cows exhibited estrus 23 d sooner (P less than .

Nucleotide sequence of the trpD and trpC genes of Salmonella typhimurium.

We have completed the nucleotide sequence determination of trpD and trpC, the second and third genes of the trp operon of Salmonella typhimurium. These genes encode two bifunctional proteins thought to have arisen by gene fusions: the trpD polypeptide contains the glutamine amido transferase and the phosphoribosyl anthranilate transferase activities, and the trpC protein possesses the N-(5'-phosphoribosyl)-anthranilic acid isomerase and the indole-3-glycerol phosphate synthetase activities. The trpD gene consists of 1593 nucleotides encoding 531 amino acids, and possesses an internal promoter (p2) located within a region from about 1400 to 1441 of the nucleotide sequence.

Initiation in vivo at the internal trp p2 promoter of Escherichia coli.

We have identified an RNA transcript initiated in vivo at the internal promoter of the Escherichia coli trp operon. The 5' end of this message overlaps the distal portion of the trpD structural gene, and the startpoint of transcription is the same as that previously determined in vitro. The relative abundance of the primary and secondary promoter transcripts in cells grown under varying conditions confirms previous genetic data suggesting that the p2 promoter is expressed at a low level, but constitutively.

The nucleotide sequence of the rho gene of E. coli K-12.

We have determined the nucleotide sequence of the rho gene which encodes the E. coli K-12 transcription termination factor. The structural gene was located on a cloned 3.

Renal handling of the monobactam azthreonam in healthy subjects.

Azthreonam is a new completely synthetic, monocyclic beta-lactam antibiotic with potent activity in vitro against most gram-negative aerobic bacteria. Its renal handling was studied in six healthy men after an intravenous loading dose of 1200 mg over 2 min followed by a continuous infusion of 500 mg/hr for 4 hr with and without oral probenecid (1 gm b.i.

Effects of photoperiod and estradiol on tonic gonadotropins in ovariectomized ewes.

Estradiol-17 beta (estradiol) inhibition of tonic (basal, as opposed to cyclic or preovulatory) serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) was evaluated in ewes subjected to long or short photoperiods. Nine mature Colombia ewes were ovariectomized early in the breeding season and implanted with either Silastic capsules containing approximately .12 g of estradiol (n = 4) or with empty capsules (n = 5).

A kinetic mechanism for the poly(C)-dependent ATPase of the Escherichia coli transcription termination protein, rho.

A study of the initial velocity kinetics of the Escherichia coli transcription termination protein rho, with respect to its poly(C)-dependent ATPase, indicates that this reaction occurs by an ordered sequential mechanism. Product inhibition and substrate analogue studies suggest that ATP binding must precede the binding of poly(C) and that the order of release of the products is ADP followed by Pi, then poly(C). A possible mechanism for relating the ATPase to the termination reaction of rho is discussed in relation to the model for rho proposed by Richardson (5).

Autotrophic picoplankton in the tropical ocean.

In phytoplankton of the eastern tropical Pacific Ocean from 25 to 90 percent of the biomass (measured as chlorophyll a) and 20 to 80 percent of the inorganic carbon fixation were attributable to particles that could pass a screen with a 1-micrometer pore diameter. Evidence is presented that these are indeed autotrophic cells and not cell fragments.