Water relations of coastal and estuarine Rhizophora mangle: xylem pressure potential and dynamics of embolism formation and repair.

Physiological traits related to water transport were studied in Rhizophora mangle (red mangrove) growing in coastal and estuarine sites in Hawaii. The magnitude of xylem pressure potential (P x), the vulnerability of xylem to cavitation, the frequency of embolized vessels in situ, and the capacity of R. mangle to repair embolized vessels were evaluated with conventional and recently developed techniques. The osmotic potential of the interstitial soil water (πsw) surrounding the roots of R. mangle was c. -2.6±5.52×10-3 and -0.4±6.13×10-3 MPa in the coastal and estuarine sites, respectively. Midday covered (non-transpiring) leaf water potentials (ΨL) determined with a pressure chamber were 0.6-0.8 MPa more positive than those of exposed, freely-transpiring leaves, and osmotic potential of the xylem sap (πx) ranged from -0.1 to -0.3 MPa. Consequently, estimated midday values of P x (calculated by subtracting πx from covered ΨL) were about 1 MPa more positive than ΨL determined on freely transpiring leaves. The differences in ΨL between covered and transpiring leaves were linearly related to the transpiration rates. The slope of this relationship was steeper for the coastal site, suggesting that the hydraulic resistance was larger in leaves of coastal R. mangle plants. This was confirmed by both hydraulic conductivity measurements on stem segments and high-pressure flowmeter studies made on excised leafy twigs. Based on two independent criteria, loss of hydraulic conductivity and proportions of gas- and liquid-filled vessels in cryo-scanning electron microscope (cryo-SEM) images, the xylem of R. mangle plants growing at the estuarine site was found to be more vulnerable to cavitation than that of plants growing at the coastal site. However, the cryo-SEM analyses suggested that cavitation occurred more readily in intact plants than in excised branches that were air-dried in the laboratory. Cryo-SEM analyses also revealed that, in both sites, the proportion of gas-filled vessels was 20-30% greater at midday than at dawn or during the late afternoon. Refilling of cavitated vessels thus occurred during the late afternoon when considerable tension was present in neighboring vessels. These results and results from pressure-volume relationships suggest that R. mangle adjusts hydraulic properties of the water-transport system, as well as the leaf osmotic potential, in concert with the environmental growing conditions.

A comparison of the soft agar and microtitre methodologies for the L5178Y tk +/- mouse lymphoma assay.

The L5178Y tk +/- mouse lymphoma assay (MLA) has been validated as a sensitive and specific test system for the detection of mutagens/clastogens. There are currently two methodologies for performing the MLA: the original soft agar procedure and the newer microtitre procedure. While both procedures are considered acceptable, a limited amount of comparative information exists for the two methods. In this report the two methods were compared with regard to: (1) spontaneous and induced mutant frequencies; (2) cloning efficiencies; and (3) colony size distributions for mutants. In addition, small and large mutant colonies from microtitre wells were rechallenged for trifluorothymidine (TFT) resistance. In a majority of the cases, cloning efficiency values were higher for the microtitre as were the spontaneous and induced mutation frequency (MF) values. Nevertheless, when responses were compared according to mutation index (fold increase over background MF) the results from the two systems were often similar. More spontaneous small colonies were observed in the microtitre assay. While colony size distribution for induced mutant colonies was compound specific, generally, more small colonies were counted in microtitre. All mutant clones that were rechallenged with TFT demonstrated resistance. Aside from the differences mentioned above, both the microtitre and the soft agar procedures appear equally capable of identifying mutagenic agents.

Improved sensitivity of the unscheduled DNA synthesis assay in primary rat hepatocytes following culture in serum-free defined media.

The unscheduled DNA synthesis (UDS) assay has been used extensively for the in vitro detection of DNA damage caused by compound exposure. However, the in vitro UDS assay has been insensitive for the detection of certain chemicals, particularly nitroaromatic compounds, that are positive in bacterial mutation assays. Recently, studies have been reported which describe alterations in the hepatocyte membrane following collagenase perfusion. Independently, a method for serum-free tissue culture has been developed which results in the up-regulation of cell surface receptors and which may restore membrane functions. Fourteen compounds, including seven nitroaromatics, were evaluated in the in vitro UDS assay employing a serum-free procedure. Five compounds that were previously reported positive in the standard in vitro UDS assay were also found positive using the serum-free method. In addition, five of the nitroaromatic compounds produced positive results with the serum-free method. 1-Methyl-3-nitro-1-nitrosoguanidine and 2-acetylaminofluorene, routinely used as positive controls in the UDS assay, showed greater activity in the serum-free assay. These results suggest that the use of serum-free media improves the sensitivity of the in vitro UDS assay and that the serum-free procedure potentially offers an effective alternative to the more labor intensive and more costly in vivo UDS assay.

Quantification of unscheduled DNA synthesis by a whole cell counting method.

A procedure was developed for the quantification of the autoradiographic assay for unscheduled DNA synthesis. Relative to commonly used practices for grain counting, this procedure provides a more accurate net nuclear grain count by eliminating the subjectivity currently associated with selection of the areas to be counted for the cytoplasmic background count. Briefly, the object area and aperture area modes of an ARTEK 880 colony counter are used to collect values for the total number of silver grains over a particular cell (nuclear and cytoplasmic counts), as well as for the nuclear and cytoplasmic areas. These values are then employed in a short algorithm to determine the net nuclear grain count. This new method provides greater sensitivity for defining weak UDS responses and the data collected readily lends itself to statistical analysis.

A new scale for the assessment of depressed mood in demented patients.

Twenty-one subjects with clinically diagnosed dementia of the Alzheimer type were rated on the Dementia Mood Assessment Scale, a new instrument intended to measure the severity of depressed mood in cognitively impaired patients. Ratings were based on direct observation and a semistructured interview of the patient. Interrater reliability was established. There were highly significant correlations between patients' scores on the instrument's 17-item depression subscale and their scores on global measures of depression and sadness. The potential usefulness of this new scale in assessing the severity of depression in demented patients longitudinally or under drug treatment conditions is discussed.

L-deprenyl in Alzheimer's disease. Preliminary evidence for behavioral change with monoamine oxidase B inhibition.

Since monoamine neurotransmitter disturbances exist in some cases of dementia of the Alzheimer's type (DAT), monoamine-enhancing drugs may ameliorate some symptoms of DAT. L-Deprenyl is a monoamine oxidase (MAO) inhibitor that is generally free of undesired effects. At low doses (10 mg/d) it selectively inhibits MAO-B, an enzyme whose level is elevated in the brains of patients with DAT who are studied post mortem. At higher doses it has more complex effects, including inhibition of MAO-A plus MAO-B. We administered 10 mg/d and 40 mg/d of L-deprenyl to 17 patients with DAT in a double-blind, placebo-controlled, serial treatment. Total Brief Psychiatric Rating Scale scores decreased significantly during 10-mg/d treatment, with decreases in measures of anxiety/depression, tension, and excitement. Approximately one half of the patients' conditions were judged to be improved clinically, with evidence of increased activity and social interaction along with reduced tension and retardation. Similar but smaller changes were observed during 40-mg/d treatment. The behavioral changes were associated with improvement in performance on a complex cognitive task requiring sustained effort. There were minimal physiologic and side effects. The greater effect of low-dose L-deprenyl therapy suggests that it is the inhibition of MAO-B, and not MAO-A, that may be important in the behavioral effects of L-deprenyl administration to patients with DAT.

On the use of a bubble formation model to calculate diving tables.

Previous decompression tables for humans were based upon unsupported assumptions because the underlying processes by which dissolved gas is liberated from blood and tissue were poorly understood. Some of those assumptions are now known to be wrong, and the recent formulation of a detailed mathematical model describing bubble nucleation has made it possible to calculate diving tables from established physical principles. To evaluate this approach, a comprehensive set of air diving tables has been developed and compared with those of the U.S. and British Navies. Conventional decompressions, altitude bends, no-stop thresholds, and saturation dives are all successfully described by one setting of four global nucleation parameters, which replace the U.S. Navy's matrices of M-values. Present air diving tables show great irregularity, even within sets created by the same authors. In contrast, this new approach is remarkably self-consistent, permitting accurate interpolation and extrapolation.

Application of bubble formation model to decompression sickness in fingerling salmon.

Recently a new cavitation model has been proposed in which bubble formation in aqueous media in initiated by spherical gas nuclei stabilized by surface-active membranes of varying gas permeability. In previous application of the varying permeability model, good agreement has been obtained with experimental limits in pressure reduction for gelatin, rats, and humans following steady-state exposures. We new extend this investigation to fingerling salmon and demonstrate that a satisfactory description of the decompression data of D' Aoust et al. (Undersea Biomed Res 1980; 7:199-209) is provided by the model with parameter values that are similar to those found for other physical and biological systems. This adds further evidence for the generally of the model as well as for the importance of bubble nucleation as the primary and controlling event in decompression sickness.

On the use of oxygen to facilitate decompression.

Oxygen is widely used at elevated partial pressures to facilitate decompression, yet the optimum dosage and the magnitude of the beneficial effects are poorly known. This is because oxygen enhancements, expressed as increases in the allowed pressure reductions, are small and easily masked by individual variation. Furthermore, oxygen can also produce detrimental results, and the range from a therapeutic to a toxic dose is narrow. Berhage and McCracken recently reported two massive investigations involving 1185 rats and 60 experimental conditions. These authors suggest that the conventional concept of an "equivalent air depth" (EAD) is untenable and that oxygen must be considered in calculating the totat tissue gas tension. We find instead that the observations of Berghage and McCracken are compatible with a model in which the tensions of oxygen and carbon dioxide dissolved in tissue are taken into account, and that this model, in turn, agrees with EAD predictions of oxygen enhancements for subtoxic oxygen pressures.

Application of a bubble formation model to decompression sickness in rats and humans.

Although decompression sickness results from bubble formation in blood or tissue, pressure schedules currently in use are essentially empirical and contain little input from cavitation theory. The recent convergence of three lines of investigation suggests that a synthesis of practice and theory may now be possible. The data consist of pressure reduction limits for gelatin, rats, and humans following steady-state exposures. From the gelatin studies, a model has been developed in which bubble formation is initiated by spherical gas nuclei stabilized by surface-active skins of varying gas permeability. We demonstrate that the model is also in good agreement with data on rats and humans over a wide range of pressures and that the model parameters assume sensible values in each case. This suggests that cavitation theory can provide a rationale for current diving practice and can serve to secure, consolidate, and extend this practice.

Stabilization of gas cavitation nuclei by surface-active compounds.

Gas bubbles are the primary agent in producing the pathogenic effects of decompression sickness. Numerous experiments indicate that bubbles originate in water, and probably also in man, as pre-existing gas nuclei. This is surprising considering that gas phases larger than 1 micron should rise to the surface of a standing liquid, whereas smaller ones should dissolve rapidly due to surface tension. Several stabilizing mechanisms have been suggested, and each has been refuted on experimental grounds. In this article, we propose a new model that arises out of a systematic study of the earlier theories. We review these theories and conclude that gas cavitation nuclei must be held intact by surface-active skins that are initially permeable. The first quantitative analysis of bubble formation data from supersaturated gelatin is summarized and leads to the further conclusion that skins can become impermeable if the ambient pressure is increased rapidly by a sufficient amount. Our model owes much to Sirotyuk, who "demonstrated experimentally that stabilization of gas bubbles acting as cavitation nuclei in water is always attributable to the presence of surface-active substances in the water".

Bubble formation within decompressed hen's eggs.

Decompression sickness follows a reduction in ambient pressure and is a result of bubble formation in blood or tissues. The origin of such bubbles is the subject of considerable controversy, and a number of mechanisms have been proposed to account for them. In testing these mechanisms, freshly-laid hen's eggs provide a particularly intriguing model--namely, an intact biological system in which bubbles form readily and many of the proposed processes are excluded.