Seawater strontium isotopes, Acid rain, and the cretaceous-tertiary boundary.

A large bolide impact at the end of the Cretaceous would have produced significant amounts of nitrogen oxides by shock heating of the atmosphere. The resulting acid precipitation would have increased continental weathering greatly and could be an explanation for the observed high ratio of strontium-87 to strontium-86 in seawater at about this time, due to the dissolution of large amounts of strontium from the continental crust. Spikes to high values in the seawater strontium isotope record at other times may reflect similar episodes.

Cubanite: A New Sulfide Phase in CI Meteorites.

Cubanite (CuFe(2)S(3)), previously unobserved in meteorites, has been discovered in two carbonaceous chondrites, Orgueil and Alais. The association of this mineral with low-copper pyrrhotite suggests that it formed in a low-temperature environment on the meteorite parent body.

East pacific rise: hot springs and geophysical experiments.

Hydrothermal vents jetting out water at 380 degrees +/- 30 degrees C have been discovered on the axis of the East Pacific Rise. The hottest waters issue from mineralized chimneys and are blackened by sulfide precipitates. These hydrothermal springs are the sites of actively forming massive sulfide mineral deposits. Cooler springs are clear to milky and support exotic benthic communities of giant tube worms, clams, and crabs similar to those found at the Galápagos spreading center. Four prototype geophysical experiments were successfully conducted in and near the vent area: seismic refraction measurements with both source (thumper) and receivers on the sea floor, on-bottom gravity measurements, in situ magnetic gradiometer measurements from the submersible Alvin over a sea-floor magnetic reversal boundary, and an active electrical sounding experiment. These high-resolution determinations of crustal properties along the spreading center were made to gain knowledge of the source of new oceanic crust and marine magnetic anomalies, the nature of the axial magma chamber, and the depth of hydrothermal circulation.

Matrix metalloproteinase-3-dependent generation of a macrophage chemoattractant in a model of herniated disc resorption.

Herniated disc (HD) is a common health problem that is resolved by surgery unless spontaneous resorption occurs. HD tissue contains abundant macrophage infiltration and high levels of matrix metalloproteinases (MMPs) MMP-3 and MMP-7. We developed a model system in which disc tissue or isolated chondrocytes from wild-type or MMP-null mice were cocultured with peritoneal macrophages and used this system to investigate the role of MMPs and chondrocyte/macrophage interactions in disc resorption. We observed a marked enhancement of MMP-3 protein and mRNA in chondrocytes after exposure to macrophages. Chondrocytic MMP-3, but not MMP-7, was required for disc resorption, as determined by assaying for a reduction in wet weight and proteoglycan content after 3 days of coculture. Surprisingly, chondrocyte MMP-3 was required for the generation of a macrophage chemoattractant and the subsequent infiltration of the disc tissue by proteolytically active macrophages. We conclude that macrophage induction of chondrocyte MMP-3 plays a major role in disc resorption by mechanisms that include the generation of a bioactive macrophage chemoattractant.

Impact of Ethylene Glycol Toxicity on Donor Organ Viability: A Case Report.

Using kidneys from deceased donors whose demise was secondary to ethylene glycol (EG) toxicity requires considerable thought and planning. The exact impact that kidneys from these donors could have is unclear. The shortage of viable organs and growing wait list mortality should lead us to consider these allografts as potential life-saving transplants. Because it is crucial for the transplant community to use every available allograft, we need to develop processes that optimize each possible scenario. This article is a discussion of the viability of kidneys from a donor with EG-induced brain death and a proposed algorithm for encouraging the use of renal allografts after EG toxicity.

The 92-kDa gelatinase B is expressed by advanced stage melanoma cells: suppression by somatic cell hybridization with early stage melanoma cells.

The production and local release of various proteolytic enzymes, either by tumor cells or tumor-associated stromal cells, is thought to facilitate the malignant behavior of solid tumors. Human cutaneous melanoma offers an excellent clinical model to study the possible contribution of such proteases to solid tumor progression because melanoma goes through a series of well defined stages in its pathogenesis; moreover, permanent cell lines have been established from these various stages. As a first step to analyzing the gelatinolytic enzymes in melanoma pathology, we examined cell lines derived from early stage primary melanomas in which patients were cured of their disease and compared the results to those obtained with cell lines established from advanced stage primary lesions or metastases (i.e., from patients who eventually succumbed to the disease). We found that 80% of cell lines examined from early stage lesions constitutively produced only the 72-kDa gelatinase A but never the 92-kDa gelatinase B. In contrast, the majority of advanced stage cell lines examined produced both the 72-kDa gelatinase A and the 92-kDa gelatinase B. Advanced stage cell lines that did not constitutively produce the 92-kDa gelatinase B could be induced to do so with transforming growth factor beta, interleukin 1 beta or 12-O-tetradecanoyl-phorbol-13-acetate. In total, 0 of 5 early stage cell lines constitutively expressed the 92-kDa gelatinase B, and only 2 of 5 could be induced to produce this activity. In contrast, all advanced stage cell lines that were evaluated either constitutively or inducibly produced the 92-kDa gelatinase B. To analyze the mechanism by which 92-kDa gelatinase B production is switched on in the advanced stage melanoma cell lines, somatic cell hybrids were constructed using an advanced stage melanoma cell line as one partner and either one of two early stage cell lines as the other. Constitutive production of the 92-kDa gelatinase B in such hybrids was lost and could not be induced in such hybrids. Coculture of the early and advanced stage cell lines failed to recapitulate what was seen after somatic hybridization, and zymographic analysis of lysates from hybrid cell lines demonstrated no 92-kDa gelatinase B activity. Reverse transcription-PCR analysis demonstrated that the loss of 92-kDa gelatinase B production occurred at the level of steady-state mRNA for the enzyme.(ABSTRACT TRUNCATED AT 400 WORDS)

Acute reversal of the enhanced insulin action in trained athletes. Association with insulin receptor changes.

We studied the effect of aerobic training and detraining on insulin-stimulated glucose disposal and on erythrocyte insulin receptor binding. Seven endurance-trained athletes were studied at 12 h, 60 h, and 7 days after cessation of training and compared with three untrained, age- and weight-matched controls. The metabolic clearance rate of glucose as measured by the euglycemic clamp technique was 15.6 +/- 1.8 ml/kg/min (mean +/- SEM) in the trained subjects 12 h after the last bout of exercise compared with 7.8 +/- 1.2 ml/kg/min in the untrained control group. When the trained subjects refrained from physical training, the metabolic clearance rate decreased to 10.1 +/- 1.0 ml/kg/min at 60 h and further to 8.5 +/- 0.5 ml/kg/min after 7 days of detraining. The percentage of specific insulin binding to young erythrocytes (density 1.089-1.092), isolated by density gradient centrifugation, decreased from 10.4 +/- 0.9 at 12 h after the last exercise to 8.1 +/- 0.7%/3 X 10(9) cells after 60 h of detraining (P less than 0.001). The decrease in insulin binding to erythrocytes was almost entirely accounted for by a decrease in the number of insulin receptors. We conclude that the increase in peripheral insulin action seen in trained athletes is rapidly reversed, possibly by a mechanism separate from other phenomena associated with chronic training. The parallel findings of decreased in vivo insulin action and decreased insulin binding in young erythrocytes suggest that modulation of in vivo insulin response by detraining may be at least partially mediated by changes in insulin receptor number.(ABSTRACT TRUNCATED AT 250 WORDS)

A direct estimation of the context effect on the efficiency of termination.

An in vitro assay in which terminating Escherichia coli ribosomes with different stop signals in the A-site compete for a limited amount of a release factor (RF1 or RF2) has been used to estimate the relative termination efficiencies at stop codons with different adjacent downstream nucleotides. The assay allows direct measurements of relative kcat/Km parameters for the productive association of release factors to ribosomes. The kcat/Km parameter is larger for UAA(U) than for UAA(C) programmed ribosomes and the difference in kcat/Km is much larger for RF2 (about 80%) than for RF1 (about 30%). These differences in the kcat/Km parameter are not affected by the addition of release factor RF3. The only discernible effect of RF3 is a considerable acceleration of RF1/2 recycling.The estimated kcat/Km parameters correlate well with the affinities of release factors for ribosomes programmed with different stop signals. These affinities were estimated from the extent of inhibition of ribosomal recycling by high concentrations of release factors in the absence of release factor RF3. The affinity for RF2 depends on the immediate downstream context of the stop codon in the translated mRNA and is about three times higher for UAA(U) than for UAA(C). The corresponding difference in affinities for RF1 is twofold. For all stop signals studied, the estimated affinity of RF2 for terminating ribosomes is much lower than that of RF1. It is also striking that the affinity of ribosomes for a chromosomally expressed RF2 is at least three times higher than for RF2 isolated from an overproducing E. coli strain.

Purification of active Escherichia coli ribosome recycling factor (RRF) from an osmo-regulated expression system.

Ribosome release factor (RRF) from Escherichia coli was overproduced from an osmo-expression vector. More than 40% of cell protein was RRF after 6 h of induction. A purification scheme is described that produced 50 mg of RRF from an initial culture of 2 L. The recycling time for ribosomes synthesising the tripeptide fMet-Phe-Leu in vitro in the absence of RF3 was reduced from 40 to 15 s by the addition of purified 1.5 microM RRF.

Immunochemical determination of the cellular content of polypeptide chain release factor RF3 in Escherichia coli.

Polypeptide chain termination in Escherichia coli is known to require two codon specific release factors, RF1 and RF2. A third factor, RF3, has been described to stimulate the termination. Earlier investigations have estimated the cellular content of factors RF1 and RF2. Two different immunological techniques for measuring the amount of RF3 per cell in crude E coli cell extracts are reported here, using a sensitive immunoblotting method and a sandwich assay by ELISA. Monoclonal murine antibodies and polyclonal rabbit antibodies were raised against extensively purified recombinant E coli RF3. The immunoblotting involves a specific monoclonal antibody (mAb), biotinylated second antibody and finally radioactive iodinated streptavidin. In the sandwich assay polyclonal antibodies are immobilised on a polystyrene surface before addition of crude cell extract; a specific mAb serves as primary antibody and an HRP-labelled anti-mouse Ig as secondary antibody. Both methods are accurate and rapid to perform. The number of RF3 molecules per cell in exponentially growing E coli cells was found to vary considerably according to the K12 strain examined and depended on the culture medium (from 20 to 500 molecules per cell), faster growth being positively correlated with the number of RF3 molecules per cell.

Use of double labeling and photo CD for morphometric analysis of injured skeletal muscle.

We used computer-assisted analysis of myofiber cross-sectional areas to measure skeletal muscle responses to injury and disease. We developed a simple, inexpensive method for measuring myofiber size in human muscle samples using Kodak photo compact discs (CDs) as the image source. The photo CD serves as a permanent image storage medium and provides a high-resolution image that can be used to detect small myofibers. The use of double labeling for dystrophin and desmin allowed positive identification of both degenerating and regenerating fibers in a single biopsy specimen.

Extrachromosomal elements of spirochetes.

The spirochetes include some important pathogenic bacteria, Treponema, Borrelia and Leptospira. The pathogeneses of these spirochetes are very diverse. In an attempt to learn more about the virulence factors among the spirochetes, their genetic organization and capacity have been studied. Structural analysis of the genome in Borrelia has shown that the genome is composed of one linear maxi-chromosome with additional linear minichromosomes as well as several supercoiled circular plasmids. Moreover, the molecular analysis of the terminal ends of one of the linear minichromosomes has revealed that this unique replicon has sequence similarities with poxviruses and particularly the viral agent of African swine fever. The presence of nucleic-acid-containing vesicles and its possible role in mediating DNA transfer between borreliae is an additional, very interesting feature of these organisms. Treponema does not contain any linear DNA, chromosomal or extrachromosomal, however molecular characterization of a 2.6-kb plasmid of Treponema denticola has been performed with the aim of establishing cloning vehicles to study the virulence properties of the genus Treponema.

Antibody response of fibrocystic patients to homologous 0-typable and 0-defective isolates of Pseudomonas aeruginosa.

Antibody titres to Pseudomonas aeruginosa of sera from 60 adult fibrocystic patients were determined in an enzyme linked immunosorbent assay (ELISA) with whole cells of homologous isolates which had been classified according to 0-antigen state by their reactivity with 0-typing antisera. Patients who were continuously colonised with Ps aeruginosa gave the highest titres: range 1500-64000 (mean 11000) and 500-48000 (mean 9000) with homologous 0-typable and 0-defective isolates, respectively. Lower titres to both varieties of isolates were obtained with recently colonised patients, and non-colonised patients gave titres with reference laboratory strains marginally above those of healthy controls. Serum titres of patients with sequential isolates were strain dependent and did not correlate with the 0-antigen state of the strain. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis of these sera and strains showed antibody binding primarily to high molecular weight 0-repeating units of lipopolysaccharide. It is concluded that the 0-antigen of the strain of Ps aeruginosa used in the ELISA test does not influence the titre obtained with fibrocystic sera, and it is recommended that serum titres should be assessed with a panel of homologous isolates from patients.

Inhibition of Pseudomonas aeruginosa from cystic fibrosis by selective media.

Pseudomonas Isolation Agar (selective agent, Irgasan, 25 mg/1) and Pseudomonas Selective Agar (selective agents, cetrimide 200mg/1 and nalidixic acid 15 mg/1) inhibited some strains of P aeruginosa from cystic fibrosis sputum but did not inhibit isolates from other sources. Of 200 cystic fibrosis isolates, 22 were inhibited by 16 mg/1 Irgasan, 45 by 8 mg/1 nalidixic acid, and 15 by 128 mg/1 cetrimide. We recommend that cystic fibrosis sputum should be cultured on selective and non-selective media to maximise the isolation of P aeruginosa.

Aldosterone and prolactin response to exercise in the heat in circumpubertal boys.

Thermoregulatory responses to exercise in the heat, especially sweating pattern, differ between children and adults. To determine whether such differences may be related to hormonal responses and to assess the possible association between this response and physical maturation, three groups of circumpubertal boys cycled at 50% of maximal O2 uptake (three 20-min bouts with 10 min of rest between bouts) in 42 degrees C at 20% relative humidity. On the basis of Tanner staging, 11 were prepubertal (PP), 12 midpubertal (MP), and 7 late pubertal (LP). Water ingestion was encouraged to minimize dehydration. Venous blood was sampled before and immediately after the session. Changes in heart rate, rectal temperature, and percent decrease in plasma volume did not differ among groups. There was no change in plasma osmolality in any of the groups. Resting testosterone concentrations were higher with increased level of physical maturity (PP = 0.4 +/- 0.1, MP = 8.2 +/- 1.9, LP = 13.8 +/- 1.2 nmol/l; P less than 0.05). In all groups, both aldosterone (ALD) and prolactin (PRL) markedly increased after exercise in the heat (ALD: PP = 161 +/- 40 vs. 1,289 +/- 263, MP = 173 +/- 47 vs. 1,245 +/- 153, LP = 250 +/- 76 vs. 1,681 +/- 400 pmol/l; PRL: PP = 8.1 +/- 1.2 vs. 24.9 +/- 4.2, MP = 8.8 +/- 1.0 vs. 22.0 +/- 8.9, LP = 8.4 +/- 0.8 vs. 39.0 +/- 3.6 micrograms/l; P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Acute changes in high-density lipoprotein cholesterol with exercise of different intensities.

The purpose of this study was to investigate the acute effects of exercise on plasma high-density lipoprotein cholesterol (HDL-C) and to determine whether the magnitude of this response would be affected by the intensity of the exercise. Twelve men (19-41 yr) ran an equivalent distance (9-12 km) on a treadmill on two separate occasions. On one occasion the exercise was performed at a speed that elicited 60% of the subject's maximal O2 uptake (VO2max), and on the other occasion exercise was performed at a speed that elicited 90% of VO2max. Changes in total cholesterol, triglycerides (TG), HDL-C, HDL apoprotein A (HDL-A), HDL saturation, lactate (LA), and free fatty acids (FFA) were measured during the course of each run, and all values were corrected for changes in plasma volume as indicated by hematocrit. There were significant increases (P less than 0.01) in HDL-C, HDL-A, and HDL saturation with exercise at both intensities, but greater increases in HDL-C (25 vs. 14%) and HDL-A (18 vs. 8%) were observed with the higher intensity exercise. Plasma FFA and TG did not differ between conditions, but LA concentrations rose significantly during the high-intensity exercise. These results indicate that increases in HDL components can occur with a relatively moderate exercise session and that the magnitude of these increases are directly related to the exercise intensity.

Influence of protein intake and training status on nitrogen balance and lean body mass.

The present study examined the effects of training status (endurance exercise or body building) on nitrogen balance, body composition, and urea excretion during periods of habitual and altered protein intakes. Experiments were performed on six elite bodybuilders, six elite endurance athletes, and six sedentary controls during a 10-day period of normal protein intake followed by a 10-day period of altered protein intake. The nitrogen balance data revealed that bodybuilders required 1.12 times and endurance athletes required 1.67 times more daily protein than sedentary controls. Lean body mass (density) was maintained in bodybuilders consuming 1.05 g protein.kg-1.day-1. Endurance athletes excreted more total daily urea than either bodybuilders or controls. We conclude that bodybuilders during habitual training require a daily protein intake only slightly greater than that for sedentary individuals in the maintenance of lean body mass and that endurance athletes require daily protein intakes greater than either bodybuilders or sedentary individuals to meet the needs of protein catabolism during exercise.

Contractile adaptations in the human triceps surae after isometric exercise.

Ultrastructural and twitch contractile characteristics of the human triceps surae were determined in seven healthy but very sedentary subjects before and after 16 wk of unilateral isometric training at 100% maximal voluntary contraction. After training, twitch contraction time decreased by approximately 20%. One-half relaxation time, peak twitch torque, and percent fiber type in any of the muscles of the triceps surae complex were not changed by training. Type I and type II fiber areas increased in the soleus by approximately 30%, but only type II fibers showed an increased in area in the lateral gastrocnemius (40%). Despite such changes in fiber area, the volume density of the sarcoplasmic reticulum-transverse tubular (SR) network averaged 3.2 +/- 0.6 and 5.9 +/- 0.9% in type I and type II fibers, respectively, before and after training in the two heads of the gastrocnemius. Type I SR fraction increased to 3.5 +/- 1.2% after training in the soleus; however, correlations were not significant between the change in the volume density of SR and the change in twitch contraction time (R = 0.46, P = 0.45) or the change in one-half relaxation time (R = -0.68, P = 0.08). The results demonstrate that isometric training at 100% maximal voluntary contraction induced changes in twitch contraction time that were not directly related to changes in the volume density of SR in fibers of the triceps surae.