Subcellular calcium transients visualized by confocal microscopy in a voltage-clamped vertebrate neuron

Confocal laser-scanned microscopy and long-wavelength calcium (Ca2+) indicators were combined to monitor both sustained and rapidly dissipating Ca2+ gradients in voltage-clamped sympathetic neurons isolated from the bullfrog. After a brief activation of voltage-dependent Ca2+ channels, Ca2+ spreads inwardly, and reaches the center of these spherical cells in about 300 milliseconds. Although the Ca2+ redistribution in the bulk of the cytosol could be accounted for with a radial diffusion model, local nonlinearities, suggesting either nonuniform Ca2+ entry or spatial buffering, could be seen. After electrical stimulation, Ca2+ signals in the nucleus were consistently larger and decayed more slowly than those in the cytosol. A similar behavior was observed when release of intracellular Ca2+ was induced by caffeine, suggesting that in both cases large responses originate from Ca2+ release sites near or within the nucleus. These results are consistent with an amplification mechanism involving Ca2(+)-induced Ca2+ release, which could be relevant to activity-dependent, Ca2(+)-regulated nuclear events.     

Differential reactivity of bovine lymphocytes to species of Brucella

The reactivity of bovine lymphocytes to 4 species of Brucella was tested in thymidine-uptake assays, using long-term cultured lymphocytes and freshly obtained blood mononuclear cells. Lymphocytes were taken from cows that had been challenge exposed with a virulent strain of B abortus at midgestation. The cows were classified retrospectively as being naturally resistant or susceptible to brucellosis. Lymphocytes taken from these cows had 3 patterns of reactivity with species of Brucella: pattern 1 was defined by reactivity with 4 species (B abortus, B canis, B suis, and B melitensis); pattern 2 was defined by reactivity with all these species, except B melitensis; pattern 3 was defined by reactivity with B abortus and B canis, but not with B suis or B melitensis. There was a statistically significant correlation between susceptibility to brucellosis and expression of lymphocyte cross-reactivity with B suis (P less than 0.01) and with B melitensis (P less than 0.001).     

A comparison of the use of different swab materials for optimal diagnosis of amoebic gill disease (AGD) in Atlantic salmon (Salmo salar L.)

Routine gill swabbing is a non-destructive sampling method used for the downstream qPCR detection and quantitation of the pathogen Neoparamoeba perurans, a causative agent of amoebic gill disease (AGD). Three commercially available swabs were compared aiming their application for timelier AGD diagnosis (Calgiswab^® (calcium alginate fibre-tipped), Isohelix^® DNA buccal and cotton wool-tipped). Calcium alginate is soluble in most sodium salts, which potentially allows the total recovery of biological material, hence a better extraction of target organisms’ DNA. Thus, this study consisted of (a) an in vitro assessment involving spiking of the swabs with known amounts of amoebae and additional assessment of retrieval efficiency of amoebae from agar plates; (b) in vivo testing by swabbing of gill arches (second, third and fourth) of AGD-infected fish. Both in vitro and in vivo experiments identified an enhanced amoeba retrieval with Calgiswab® and Isohelix® swabs in comparison with cotton swabs. Additionally, the third and fourth gill arches presented significantly higher amoebic loads compared to the second gill arch. Results suggest that limiting routine gill swabbing to one or two arches, instead of all, could likely lead to reduced stress-related effects incurred by handling and sampling and a timelier diagnosis of AGD.     

Growth,physiology, and nutrition of loblolly pine seedlings stressed by ozone and acidic precipitation: A summary of the ropis-south project

Previously published results from a multidisciplinary research program, Response of Plants to Interacting Stress (ROPIS), initiated by the Electric Power Research Insitute are summarized here. The overall objective of the ROPIS program was to develop a general mechanistic theory of plant response to air pollutants and other stresses. Direct and indirect phytotoxic impacts of O₃ combined with induced deficiencies of key nutrients as a consequence of acidic deposition are important components in many of the hypotheses used to explain reported declines in forest growth. In order to address these concerns as they relate to loblolly pine (Pinus taeda L.) growth and develop a greater level of mechanistic understanding of stress response, a study was formulated with two major objectives: (i) over a multi-yr period evaluate the role of loblolly pine genotype in governing loblolly growth response to O₃; and (ii) determine the underlying physiological and edaphic basis for loblolly growth response to O₃, acidic precipitation, and soil Mg status. An open-top chamber facility located at Oak Ridge, TN provided controlled O₃ exposure for the genotype screening study (1986–88) and controlled O₃ exposure and rainfall exclusion and addition for the O₃-rainfall acidity-soil Mg interaction study (1987–89). A variety of experimental techniques, measurements, and statistical procedures were used over a 4-yr period to quantify various aspects of plant growth, physiology, and soil-plant relationships. Results from the genotype screening study indicate that although family-specific O₃ effects were observed at the end of the first year, no statistically significant O₃ effects on diameter, height, or total biomass were evident at the end of three growing seasons; nor were any significant O₃-family interactions found. In the interaction study, rainfall acidity and soil Mg level had only minimal affects on seedling growth and physiology. Ozone exposure produced significant changes in many variables, the most important being a net retention of carbon in above-ground biomass and a subsequent reduction in carbon allocation to the root system. This change could have important longterm implications for the tree's ability to obtain water and nutrients, maintain important rhizosphere organisms, and achieve a level of vigor that protects against disease and insect attack.     

Assessment of changes in blood volume in response to resuscitative fluid administration in dogs

Objective: To determine the continuous changes in blood volume in response to fluid administration using an in‐line hematocrit monitor. Design: Prospective study. Setting: Research laboratory. Animals: Four healthy dogs. Interventions: Each dog received intravenous boluses of 80 mL/kg of 0.9% saline (S), 4 mL/kg of 7.5% saline (HS), 20 mL/kg of dextran 70 (D), 20 mL/kg of hetastarch (HES), or no fluids (control, C) on separate occasions. Fluids were administered at 150 mL/min in the S, D, and HES groups, and at 1 mL/kg/min in the HS group. Measurements and main results: Blood volume changes were measured every 20 seconds for 240 minutes using an in‐line hematocrit monitor. There was a rapid rise in blood volume during all infusions. Immediately after the administration of crystalloid fluids, the rapid rise in blood volume ceased. Subsequently, there was a steep decline in blood volume for 10 minutes, and a slower decline thereafter. In contrast, the rise in blood volume continued for at least 10 minutes after the infusion of the colloids was complete, and a plateau was observed for the remainder of the experiment. The blood volume effect, as measured by area under the curve, was significantly greater in the saline group than the other groups during the infusion time and for the 0–240 minutes time period. The areas under the curve for the two colloid solutions were not significantly different from each other during any time periods. The percent increase in blood volume immediately following the infusions was 76.4±10.0 in the S group, 17.1±3.2 in the HS group, 23.0±10.5 in the D group, and 27.2±6.4 in the HES group. At 30 minutes from the start of the infusion, the mean percent increases in blood volumes were 35.2±9.3 in the S group, 12.3±0.9 in the HS group, 35.9±7.3 in the D group, and 36.8±6.5 in the HES group. At 240 h post‐infusion, the mean percent increases in blood volume were 18.0±9.7 in the S group, 2.9±6.1 in the HS group, 25.6±16.1 in the D group, and 26.6±8.6 in the HES group. The C group had a mean percent change in blood volume of ?3.7±3.4 at the end of the experiment. Conclusions: This study indicates that the rapid administration of saline at clinically relevant doses leads to the largest immediate increase in blood volume, although this change is transient because of rapid redistribution of the fluid. Despite a brief increase in blood volume that was almost 3 times the volume administered, hypertonic saline led to the smallest increase in blood volume post‐infusion. The synthetic colloid solutions increased the blood volume by an amount greater than that infused and the effect was sustained for a longer period of time than seen following crystalloid administration, but the maximum increase in blood volume was significantly less than saline. The measurement of continuous changes in blood volume, using an in‐line hematocrit monitor, was a useful means of assessing the dynamic effects of fluid administration in dogs in a research setting.     

Porphyric insecticides. IV: Structure-activity study of substituted phenanthrolines

The porphyric-insecticide-modulating activities of 1,10-phenanthroline and eight of its analogs were investigated. The insecticidal efficacy of these compounds was closely associated with their ability to enhance the conversion of exogenous δ-aminolevulinic acid (ALA) to protoporphyrin IX (Proto). As observed for photodynamic herbicidal effects in plants, the presence of nitrogen atoms at positions 1 and 10 of the macrocycle was essential for porphyric insecticidal activity. This was evidenced by the very limited activity of phenanthrene, in which positions 1 and 10 are occupied by carbon instead of nitrogen atoms. On the other hand, enhancement of Proto formation and porphyric insecticidal activity were maintained following methyl, chloro and nitro group substitution at the periphery of 1,10-phenanthroline. In contrast, Proto levels and photodynamic toxicity were reduced by hydroxy and phenyl substitution at the same positions. Benzyl substitution at the 2–3 and 8–9 positions was also inhibitory. Quantitative structure-activity calculations suggested a relationship between peripheral group substitution and physicochemical properties of the substituted compounds. Electron density changes in 1,10-phenanthroline and its analogs that appeared to be related to reduced efficacy included (a) appearance of positive charge-binding volumes at positions 4 and 7 of the 1,10-phenanthroline macrocycle, which flanks positive charge-repelling volumes, (b) a dramatic increase in superdelocalisability (i.e. electron density) over some unoccupied molecular orbitals, and (c) electronic charge at positions 1 and 10 of the macrocycle. Large increases in van der Waals volumes also exerted negative effects on insecticidal efficacy.