Effects of 2-formylpyridine monothiosemicarbazonato copper II on red cell components

2-Formylpyridine monothiosemicarbazonato copper II (CuL⁺) is readily taken up by red cells and is initially bound to glutathione and hemoglobin. Glutathione was depleted within 5 hr of incubation, presumably by oxidation mediated by CuL⁺ and O₂ with concomittant generation of toxic oxygen species. Cupric ion was slowly transferred from CuL⁺ to hemoglobin within about 7 hr, and hemoglobin was oxidized until the major form prevailing after 10 hr was α₂β₂⁺. Little increase in hemolysis due to addition of CuL⁺ dissolved in the radical scavenger dimethyl sulfoxide was observed with prolonged incubation. Strong inhibition of red cell hexokinase by CuL⁺ was observed when the enzymes in red cell lysates and hemoglobin-free red cell lysates were examined. CuL⁺ was also an effective inhibitor of yeast hexokinase. However, the inhibitory effect of CuL ⁺ within the red cells was less pronounced. It is suggested that even though intracellular accumulation of CuL ⁺ creates an oxidizing environment and is potentially capable of inhibiting thiol enzymes such as hexokinase, protective effects are exerted in the red cell by the presence of hemoglobin, of radical scavengers, and of high levels of enzymes that detoxify toxic oxygen species. Address reprint requests to Dr. W.E. Antholine, Department of Radiology, or Dr. F. Taketa, Department of Bio     

Estrogen - Induced release of luteinizing hormone in prepubertal and postpubertal heifers

This experiment was designed to determine the age at which estradiol-17beta (E(2)) first induces a preovulatory-like surge of luteinizing hormone (LH) in prepubertal heifers. Responses of prepubertal animals 3 to 4 and 5 to 6 months of age were compared with those of postpubertal heifers that received 25 mg prostaglandin F(2)alpha at 0800 hr on day 15 of the estrous cycle. E(2) (500mug) induced surges of LH in 1 5 heifers 3 to 4 months of age, 3 3 heifers 5 to 6 months of age and 5 5 postpubertal heifers. Duration of response and interval between E(2) injection and peak of the response were longer in postpubertal heifers than in those 5 to 6 months old (P<0.10). Peak response and total amount of LH released were greater in animals 5 to 6 months old (P<0.10). Only one prepubertal heifer had elevated concentrations of progesterone following an LH surge. Four of 5 postpubertal heifers receiving E(2) and 3 of 4 postpubertal heifers receiving corn oil had corpora lutea and similar patterns of progesterone concentrations. We conclude that ability to release an LH surge in response to E(2) develops in heifers between 3 and 5 months of age, but that this induced surge does not cause ovulation.     

Survival mechanisms of the central mudminnow (Umbra limi), fathead minnow (Pimephales promelas) and brook stickleback (Culaea inconstans) for low oxygen in winter

Synopsis Three species (central mudminnow, fathead minnow and brook stickleback) survive when a northern Wisconsin lake becomes anoxic in winter. Some gas bubbles beneath the ice contained as much as 11° oxygen when the lake water contained <0.30 mg 1^–1 dissolved oxygen. Experiments conducted in the field determined that gas bubbles prolonged survival of all species, especially the mudminnow and stickleback. In the laboratory, brook sticklebacks exhibited the lowest and fathead minnows the highest routine metabolic rate corrected for weight. Rate of gill ventilation of all three increased from 20 to 70 beats per minute as oxygen levels declined from 4.0 to 0.25 mg 1^–1. At low oxygen levels they moved to the upper one-third of the test tanks. Small size, low metabolic rate, tolerance of low oxygen conditions and reduced activity resulted in reduced demand for dissolved oxygen. Head shape, ventilation rate, vertical movement and utilization of high oxygen microzones also enhanced exploitation of low levels of dissolved oxygen. Central mudminnows used oxygen directly from gas bubbles found under the ice.     

Resonance Raman studies on 8-mercaptoriboflavin

Resonance Raman (RR) spectroscopy on various ionic species of 8-mercaptoriboflavin in aqueous solution shows large vibrational spectral changes upon ionization of the 8-SH group. These changes and the superposition of the RR excitation profile and the visible spectrum for the 8-S^? species indicate that there is a substantial amount of the “quinoid thioketone” resonance form present in aqueous solutions of the 8-S^? mercaptoriboflavin. RR spectra of 8-mercaptoriboflavin bound to riboflavin binding protein confirm the conclusion (reached on the basis of visible spectra) that the 8-mercaptoriboflavin binds to the protein in the 8-SH, protonated, form. There are, however, changes in the 1250-cm^?1 region of the RR spectrum upon binding aqueous 8-mercaptoflavin to the protein. The 1257-cm^?1 band in aqueous solution moves to 1248 cm^?1 on the protein surface. This shift is also observed in DMSO solution of 8-mercaptoriboflavin. Since the 1257-cm^?1 band shifts upon formation of the N-3 deuteroriboflavin, we interpret this lowering of a δ-H mode at N-3 to mean that there is weaker hydrogen bonding between flavin N-3 and riboflavin binding protein than between flavin and water.     

Comparison of nets and pump sampling gears to assess zooplankton vertical distribution in stratified lakes

The efficiency of a cantilevered bridle net was tested in comparisonwith a Wisconsin net and a pumping system to sample zooplanktonorganisms in three water layers (epi-, meta- and hypolimnion)of three Canadian Shield lakes. Variations among samplers werecompared to variations due to within-lake vertical distributionof zooplankters and among lake variations. For each lake andwater layer, we also assessed the efficiency of the three methodsaccording to the catches of zooplanktonic taxa. The highestpercentages of variation were generally due to lake or water-layereffects; interaction between sampling gears and water layerswas above 50% for most taxa, except cladoceran. Sampling methodsexplained more variation than the lake effect for some zooplanktontaxa, indicating that using different sampling devices couldpotentially alter the among-lake variation interpretation ofzooplankton abundance. The pumping system captured higher densitiesof animals per taxa than the cantilever and the Wisconsin nets.The cantilever net generally captured mobile taxa more efficiently(Polyarthra vulgaris, copepods, Daphnia sp., Diaphanosoma brachyurumand chaoborids) than the Wisconsin net and the pumping system,but its efficiency varied among water layers.     

Aggregation and spawning by lampreys (genus Ichthyomyzon) beneath cover

Synopsis Lampreys are generally reported to spawn in shallow water on open, gravel bottoms. During surveys in Wisconsin and Minnesota, we regularly observed aggregations of adult Ichthyomyzon c.f. gagei, I. castaneus, and I. fossor beneath such cover objects as boulders, woody debris, and, at one site, vegetation. In some cases, observations of eggs or rapid quivering by individual lampreys indicated that spawning was occurring. The literature includes scattered anecdotal reports of similar behavior in other populations of Ichthyomyzon. Our data for I. c.f. gagei suggest that aggregations beneath cover objects occur at a greater range of depths than those in the open, but that aggregations in the open can contain greater numbers of individuals. Facultative spawning beneath cover objects may permit lampreys to spawn in deep waters with swift current where spawning could not otherwise occur. Moreover, this behavior may reduce the vulnerability of spawning lampreys to some types of predators.     

Inhibitory effect on limb morphogenesis by cells of the polarizing zone coaggregated with pre- or postaxial wing bud mesoderm.

The influence of cells of the polarizing zone mesoderm on the morphogenesis of recombinant chick limbs was studied. The recombinant buds were composed of leg bud ectoderm and different regions of the wing bud mesoderm, which had been dissociated and reaggregated. In any case where the polarizing zone mesoderm was coaggregated with the wing mesoderm the morphogenetic capabilities of the recombinant were reduced. This was the case with postaxial mesoderm, preaxial mesoderm plus polarizing tissue, and postaxial mesoderm from which a piece of the nonpolarizing mesoderm (comparable in size to the polarizing zone) had been removed. All of these gave outgrowths with digits in only a very low percentage of cases. In contrast, those recombinants without polarizing mesoderm developed outgrowths with digits in a high percentage of cases, indicating good morphogenesis. Finally, if the polarizing zone were removed prior to dissociation, the recombinant limb, composed of the total remaining wing bud mesoderm plus leg bud ectoderm, exhibited a higher percentage of complete morphogenesis than if the polarizing zone had been part of the recombinant.It is clear that cells of the polarizing zone, when dissociated, and coaggregated with wing mesoderm, are inhibitory to the morphogenetic performance of that mesoderm in the recombinant limb situation.     

Wisconsin desmids. III. Desmid community composition and distribution in relation to lake type and water chemistry

This investigation summarizes quantitative data on the generic composition of the euplankton and aufwuchs desmid communities of 61 Wisconsin lakes, and analyzes the information with respect to 1) the role of the various genera in terms of frequency, density, and relative importance, 2) the suitability of various lake types for harboring desmid communities, and 3) the relationships between chemical parameters and desmid distribution. The generaStaurastrum, Cosmarium, andClosterium are of wide occurrence, appear to play major roles in the communities of all lake types, and are the most tolerant of varying chemical conditions. Most euplankton genera are of importance only in acid bogs, but aufwuchs genera generally are more widely distributed. Both the euplankton and the aufwuchs communities appear to be composed of 1–4 desmid assemblages, each with a differing range of importance values. Based on biological criteria, acid bogs appear to be the most suitable lake type for harboring desmid communities and calcareous spring ponds the least suitable type. High generic diversity of desmids appears to be correlated with low conductivity, calcium and alkalinity levels, pH values of 5.1–7.0, and the presence of free CO₂. The evidence attending various hypotheses concerning water chemistry and desmid distribution appears contradictory, and further studies are needed to help clarify the situation.     

Acetylcholine stimulation of selective increases in stearic and arachidonic acids in phosphatidic acid in mouse pancreas.

During the acetylcholine-stimulated loss of phosphatidylinositol and gain in the level of phosphatidic acid in mouse pancreas, there is a selective increase in stearic and arachidonic acids in phosphatidic acid. The amounts parallel the decrease in phosphatidylinositol, which contains predominantly these two fatty acids. Addition of atropine to stimulated tissue reverses the changes. There is a selective disappearance of the stearoyl, arachidonoyl phosphatidic acid, and phosphatidylinositol increases. The changes support the hypothesis that the 1-stearoyl, 2-arachidonoyl diglyceride backbone of phosphatidylinositol becomes phosphatidic acid during acetylcholine stimulation, and is transformed back to phosphatidylinositol on reversion to the unstimulated state.