Differences in the onset of laying between island and mainland Mediterranean Blue Tits Parus caeruleus: phenotypic plasticity or genetic differences?

Blue tits Parus caeruleus breeding in deciduous or in evergreen woodlands on the mainland of Europe start to lay 3–4 weeks earlier than Blue Tits on the island of Corsica breeding in evergreen woodlands. A similar difference in average laying date between the two Blue Tit populations has been found in a relatively small sample of captive birds held in outdoor aviaries on the mainland. To provide data on the habitat contrast in Corsica for comparison with that on the mainland, a new study area was started in one of the few deciduous woodlands on Corsica. In addition, new data on laying dates of captive Blue Tits were gathered to increase the sample size of independent pairs in aviaries. This study shows that the habitat effect (deciduous ν evergreen woodland) on the average laying date of Blue Tits is relatively small in comparison with the geography effect (European mainland ν Corsica). The laying date differences between mainland and insular Blue Tits persisted whatever the type of habitat (deciduous, evergreen or outdoor aviaries on the mainland). The data support the hypothesis that the observed differences in the onset of laying between the mainland and insular Blue Tits were mainly influenced by genetic effects and relatively little by non-genetic maternal effects of by genotype—environment interactions.     

Purification of dihydropterin reductase using immobilized Cibacron Blue.

Chromatography on columns of immobilized Cibacron Blue (Blue Dextran--agarose) can be used as a major step in the purification of quinonoid dihydropterin reductase. The reductase has been isolated from fractions of beef kidney by selective binding to the immobilized Cibacron in the presence of tetrahydropterin. The binding of the reductase to Blue Dextran and its specific elution from columns of Blue Dextran--agarose indicate that the reductase possesses the dinucleotide (NAD+) binding domain. The results of kinetic experiments give validity to both our affinity chromatography of the reductase and to an ordered mechanism for the formation of tetrahydropterin. Chromatography on Blue Dextran--agarose has been used to show that folate or amethopterin can compete with Cibacron Blue for the dinucleotide domain of the reductase. The p-aminobenzoyl-glutamate moiety of the folates competes with Cibacron Blue for the NADH site of the reductase. A stable binary complex of dihydropterin reductase with NADH has been detected by gel electrophoresis.     

Intermediate electron-acceptors in quantitative cytochemistry

Summary The efficacy of Meldola Blue (MB), a new intermediate electronacceptor, has been compared with that of phenazine methosulphate (PMS) in the assay of oxidoreductase activity in cryostat sections; various tetrazolium salts have been used as the final electron-acceptors. Three enzymes: succinate dehydrogenase, glucose 6-phosphate dehydrogenase and lactate dehydrogenase were investigated, the activity in sections being quantitated by scanning and integrating microdensitometry. Phenazine methosulphate was superior to Meldola Blue in transferring reducing equivalents from reduced coenzyme to all the tetrazolium salts examined.     

Alveolar septal folding and lung inflation history.

On the basis of microscopic appearance of excised lungs, it has been thought that alveolar septa may fold and unfold during deflation and inflation. We suspected that this appearance might depend heavily on the inflation history of the lung preparation. We therefore studied, by light and electron microscopy, dog, rabbit, and rat lungs fixed over a range of inflation pressures and after a variety of inflation histories. Septal folding, as suggested by the configurations of the air spaces, by the placement of the fine and coarse connective tissue elements, and by the pattern of infolding of alveolar epithelium, was readily seen with some inflation protocols but was absent with others. Pressure at fixation was not as important as events before fixation; deflation to 3 cmH2O did not induce folding, and inflation to 16 cmH2O did not undo the folds. This range corresponds with concepts of critical opening and closing pressures. We suggest that folds form de novo during experimental preparation; one need not postulate that septal folding was present in vivo.     

The composition of stains produced by the oxidation of Methylene Blue

Synopsis The composition of some stains produced by the oxidation of Methylene Blue has been studied by thin-layer chromatography. Various named methods for the production of Polychrome Methylene Blue, Azure A, Azure B, Azure C and Methylene Violet Bernthsen have been found to give complex mixtures of varying proporitions of up to eleven dyes. Ten of these, namely Methylene Blue, Azure B, Azure A,sym-Dimethylthionine, Azure C, Thionine, Methylene Violet Bernthsen, Methyl Thionoline, Thionoline and Thionol, have been identified by their visible absorption spectra. The remaining dye could not be identified. When used on a laboratory scale, these methods give stains of constant composition independent of the batch of Methylene Blue. Stain composition as revealed in the present study has been compared with that previously indicated by other, less effective, analytical techniques. Reasons are presented why the latter give equivocal results.     

Blue Data Books - an encouraging new instrument for restoration and conservation

Abstract. In order to stress successes in restoration and conservation of species, regionally, nationally or globally, a new instrument has been developed, the ‘Blue Lists’, ‘registers of those Red List species that show a durable overall stabilization or increase of abundance in the region (nation, world) considered’. ‘Blue Data Books’ include additional information on the ecology, conservation, and promotion of the species. For describing the overall change of abundance of every individual species six categories are defined. Furthermore, the effects of nature conservation techniques (NCT) on a species are evaluated using six additional categories. In a test area of 3 431 km² in northern Switzerland, information was compiled on the change in abundance of the 708 Red List species of higher plants (spermatophyta) over the last 10-15 yr. Overall, 33% of these species showed a stabilization or even increase in abundance; these are Blue List species; ca.20% showed a decline and for almost 50% the change in abundance is not known. NCTs have been successfully applied to more than 50 % of all the species, at least locally. For about a further 30%, the required NCTs are known but have not yet been tried out. If these techniques were applied on a large scale, the decline of very many species in the test area could be stopped.     

Advanced breeding dates in relation to recent climate warming in a Mediterranean montane population of Blue Tits Cyanistes caeruleus

Trends in the onset of breeding, clutch size and numbers of hatchlings and fledglings are examined for a Mediterranean montane population of Blue Tits (Cyanistes caeruleus) subject to recent warming in springtime monitored during 20 years. Blue Tits advanced their breeding dates in relation to mean air temperatures in April and, as a consequence, laid larger clutches. However, increases in the numbers of hatchlings and fledged young over time were not statistically significant after accounting for variables of influence. The entire breeding season seems to have been displaced towards earlier dates by adjusting breeding time to increased temperatures in prebreeding time, to which Blue Tits have been more responsive than Pied Flycatchers (Ficedula hypoleuca) in the same area. The alternative hypothesis, that interference competition with Pied Flycatchers for nestboxes and caterpillars, the main common food base of nestlings, has been the driving force behind the advancement of laying of the Blue Tit population, was not supported. However, the significant advance of breeding dates in Blue Tits has not been sufficient to overcome the precipitous decline in reproductive fitness with the advancement of the season.     

Effect of Cibacron Blue F3GA on phosphoglycerate kinase of Lactobacillus plantarum and phosphoglycerate mutase of Leuconostoc dextranicum

Blue dextran or Cibacron Blue F3GA has been shown to inhibit yeast phosphoglycerate kinase [EC 2.7.2.3] competitively with respect to ATP (Thompson et al. (1975) Proc. Natl. Acad. Sci. U.S. 72, 663--667; Beissner and Rudolph (1979) J. Biol. Chem. 254, 6273--6277). However, we have found that phosphoglycerate kinase of Lactobacillus plantarum was inhibited by Cibacron Blue F3GA, the blue chromophore of blue dextran, noncompetitively with respect to ATP, but competitively with respect to 3-phosphoglycerate. Further inhibition studies with Cibacron Blue F3GA suggest that one molecule of the dye was bound per molecule of phosphoglycerate kinase at a saturated level of either substrate, but two molecules of the dye were bound per molecule of the kinase with an unsaturated level of either substrate used as a fixed substrate. Furthermore, phosphoglycerate mutase [EC 2.7.5.3] of Leuconostoc dextranicum was also inhibited by Cibacron Blue F3GA competitively with respect to 3-phosphoglycerate and noncompetitively with respect to 2,3-bisphosphoglycerate. These results suggest that the 3-phosphoglycerate-binding site on both phosphoglycerate kinase and phosphoglycerate mutase can interact with Cibacron Blue F3GA.     

A more efficient Big Blue protocol improves transgene rescue and accuracy in a adduct and mutation measurement

Transgenic mutational systems have provided researchers with an invaluable tool, allowing the measurement of both spontaneous and induced mutations. The Big Blue transgenic rodent mutagenesis system developed by Stratagene (La Jolla, CA) uses a lambda shuttle vector carrying lacI as the mutational target gene. A common criticism of the Big Blue system is that it relies on visual screening to detect mutants rather than positive selection, which is employed in more recently developed systems. The lack of positive selection, however, has provided the Big Blue system with a unique advantage, as it allows for the dynamic quantification of mutation fixation, repair, and adduct stability, since both pre-mutagenic DNA adducts and mutations can readily be quantified [Proc. Natl. Acad. Sci. U.S.A. 97 (2000) 11391]. Improvements to the standard Big Blue assay protocol are required for the visualization of mutant plaques resulting from pre-mutagenic damage, as these can appear much lighter in color than the lightest color control mutant (CM0). This increase in detection has been achieved by the development of a protocol that now permits the effective measurement of repair and mutation fixation utilizing the Big Blue system. This new protocol has also addressed efficiency, allowing for a two-fold increase in the number of plaques produced per packaging reaction and a decrease in both phage migration and plaque size, permitting a greater than three-fold increase in plating density. The implementation of this protocol will make the Big Blue assay more economical and less demanding than before, while providing researchers with an efficient means to measure both repair and mutation in this system.     

Evaluation of a new medium for the enumeration of total coliforms and Escherichia coli in Japanese surface waters

Aim: A new medium, EC‐Blue‐10, containing chromogenic and fluorogenic substrates, KNO₃ and sodium pyruvate has been developed for the rapid simultaneous detection and enumeration of total coliforms and Escherichia coli in water. Methods and Results:  Two evaluations of EC‐Blue‐10 were carried out. Firstly, EC‐Blue‐10 was compared with Colilert‐MPN for 96 water samples using MPN for total coliforms and E. coli. Secondly, the detection of coliforms and E. coli were compared using 2400 tubes of EC‐Blue‐10 and Colilert‐MPN. The regression coefficients between EC‐Blue‐10 and Colilert‐MPN for total coliforms and E. coli were 0·91 and 0·89, respectively. For the detection results, the Cohen’s kappa values between the two media were 0·79 for coliforms and 0·72 for E. coli. Conclusions: EC‐Blue‐10 is almost same as Colilert‐MPN for the detection of coliforms and E. coli in surface waters. Further evaluation for EC‐Blue‐10 is needed to verify in different geographical areas. Significance and Impact of the Study: EC‐Blue‐10 is useful method for the rapid and simultaneous detection of total coliforms and E. coli in water sample.     

Pulseless electrical activity during electroconvulsive therapy: a case report

Background

Arrhythmias resulting in cardiac arrest during electroconvulsive therapy have been reported. Most reported cases of cardiac arrest had asystole as the initial rhythm. Pulseless electrical activity as an initial rhythm of cardiac arrest during electroconvulsive therapy has never been reported. Also, thromboembolism after inflation of pneumatic tourniquet during lower limb surgery has been reported but never following tourniquet inflation during an electroconvulsive therapy.

Case presentation

We report a case involving an 81- year- old female who presented to us for an electroconvulsive therapy for severe depression and developed pulseless electrical activity immediately after electroconvulsive therapy. She was successfully resuscitated and was later found to have bilateral pulmonary emboli with a complete occlusion of the right lower lobe pulmonary artery. The source of embolus was from her left lower extremity deep venous thrombus, which we believe, got dislodged intraoperatively after inflation of pneumatic tourniquet. Our patient not only survived the massive pulmonary embolus, but also showed significant improvement in her mental status compared to her pre-admission level at the time of discharge to a sub-acute rehabilitation centre.

Conclusion

We recommend that patients who are elderly and at high risk of thromboembolism should selectively undergo a preoperative doppler ultrasound for deep venous thrombosis. Also, selective application of tourniquet in the upper limb, to monitor for seizure activity, would reduce the incidence of pulmonary thrombo-embolism as embolic events are significantly less from deep venous thromboses of upper extremities when compared to lower extremities.     

Flow cytometric determination of basophils in whole blood with n-propyl Astra Blue iodide

Within the past year, it has become apparent, in connection with its use on automatic flow cytometers, that the quality of commercially available Alcian Blue has significantly declined. A homologous series of alkylated (C1-C7) Astra Blue quaternary ammonium halides was prepared, characterized, and evaluated for the detection of basophils in whole blood. On the Technicon H6000 flow cytometer, the resolution of the basophil cluster from the main population of unstained white blood cells was found to depend on the chain length of the quaternizing alkyl group. Optimal basophil resolution was observed for the n-propyl derivative. Correlation of the new method vs Alcian Blue as the reference on the H6000 was expressed as follows: %Baso (Astra Blue) = 0.89% Baso (Alcian Blue) + 0.12% for 180 fresh whole blood samples. Within-run precision at a basophil differential count of 0.73% was characterized by SD = 0.11, identical to that obtained for Alcian Blue. Aqueous solutions of n-propyl Astra Blue iodide, in contrast to Alcian Blue, are thermally stable. Heating the reagent for 1 h at 100 degrees C did not alter solubility or cytochemical behavior. In contrast, parallel treatment of Alcian Blue yielded insoluble material by hydrolysis of the isothiouronium groups. The reagent for basophil detection comprises n-propyl Astra Blue iodide, lanthanum chloride, sodium chloride, Tween 20, and cetylpyridinium chloride. The Astra Blue derivatives were characterized by uv-vis, ir, percentage halide, paper chromatography, and 13C NMR.     

Microwave Staining of Intestinal Whole-mount Preparations with Cuprolinic Blue

Encouraged by the knowledge that microwaves have a beneficial effect on immunohistochemical reactions, the present study aimed to find out whether microwaves could improve the Cuprolinic Blue staining of enteric neurons as well as the actual method that has been developed for gastrointestinal whole-mount preparations. In addition to incorporating a microwave application in the method described by Holst and Powley (1995), some other modifications were made: two incubations before incubation in the staining solution and free-floating incubations. In the whole-mount preparations, most, if not all, enteric neurons were stained by Cuprolinic Blue. These neurons appeared as blue–green cells with non-reacting nuclei and neuronal processes. At higher magnification, the cytoplasm was characterized by a fine blue– green granulation, and the nucleolus in the nucleus appeared as a blue iridescent structure. Non-specific staining occurred in fibrocy tes and epithelial cells but, because of their location and appearance, they could easily be distinguished from neurons. The modified incubations and the incorporation of a microwave application into the conventional Cuprolinic Blue staining method turn the method into an easy-to-use one that seems to visualize most, if not all, enteric neurons in whole-mount preparations of the pig jejunum.     

DNA interaction, cytotoxicity, and radiosensitization with PtCl4(Nile Blue)2 and PtCl4(Neutral Red)2

Complexes of the positively charged, nuclear staining, quinone-imine dyes Nile Blue and Neutral Red with negatively charged tetrachloroplatinum (II) have been prepared in an effort to form neutral drugs which could gain ready access to the cellular nucleus and deliver significant quantities of the reactive tetrachloroplatinum anion to the vicinity of the DNA. Elemental analysis showed that both the Nile Blue and Neutral Red complexes with tetrachloroplatinum (II) comprised 2 mol of dye and 1 mol of tetrachloroplatinum, forming Pt(Nile Blue)2 and Pt(Neutral Red)2. Exposure of superhelical pBR322 DNA to the complexes or the dyes for 24 h followed by agarose gel electrophoresis showed that Neutral Red and Pt(Neutral Red)2 had little effect on DNA conformation, but that both Nile Blue and Pt(Nile Blue)2 could produce single-strand DNA breaks in a dose-dependent fashion. Studies in exponentially growing asynchronous, hypoxic, and normally oxygenated EMT6 cells at normal pH (7.40) and pH 6.45 demonstrated that neither dye was highly toxic, but that both complexes were capable of producing significant cytotoxicity. Both complexes killed normally oxygenated cells more efficiently than hypoxic cells, but Pt(Neutral Red)2 was more cytotoxic at pH 6.45, while Pt(Nile Blue)2 killed significantly more cells at normal pH. Both complexes decreased the survival of hypoxic EMT6 cells as indicated by the slope of the radiation survival curve [dose modifying factor (DMF) 2.90 for Pt(Nile Blue)2 and 1.45 for Pt(Neutral Red)2]. Studies with the FSaIIC murine tumor showed that both complexes were active radiosensitizing agents in vivo [DMF 1.76 for Pt(Nile Blue)2 and 1.25 for Pt(Neutral Red)2]. These results indicate that these new platinum complexes have characteristics which may make them and similar complexes effective radiosensitizing agents in humans.     

Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity.

We show here the identity of Alamar Blue as resazurin. The 'resazurin reduction test' has been used for about 50 years to monitor bacterial and yeast contamination of milk, and also for assessing semen quality. Resazurin (blue and nonfluorescent) is reduced to resorufin (pink and highly fluorescent) which is further reduced to hydroresorufin (uncoloured and nonfluorescent). It is still not known how this reduction occurs, intracellularly via enzyme activity or in the medium as a chemical reaction, although the reduced fluorescent form of Alamar Blue was found in the cytoplasm and of living cells nucleus of dead cells. Recently, the dye has gained popularity as a very simple and versatile way of measuring cell proliferation and cytotoxicity. This dye presents numerous advantages over other cytotoxicity or proliferation tests but we observed several drawbacks to the routine use of Alamar Blue. Tests with several toxicants in different cell lines and rat primary hepatocytes have shown accumulation of the fluorescent product of Alamar Blue in the medium which could lead to an overestimation of cell population. Also, the extensive reduction of Alamar Blue by metabolically active cells led to a final nonfluorescent product, and hence an underestimation of cellular activity.     

Stomatal Opening Mechanism of CAM Plants

Stomata usually open when leaves are transferred from darkness to light. However, reverse-phase stomatal opening in succulent plants has been known. CAM plants such as cacti and Opuntia ficus–indica achieve their high water use efficiency by opening their stomata during the cool, desert nights and closing them during the hot, dry days. Signal transduction pathway for stomatal opening by blue light photoreceptors including phototropins and the carotenoid pigment zeaxanthin has been suggested. Blue light regulated signal transduction pathway on stomatal opening could not be applied to CAM plants, but the most possible theory for a nocturnal response of stomata in CAM plants is photoperiodic circadian rhythm.     

Maintaining Critical Habitat in a Heavily Managed Landscape: Effects of Power Line Corridor Management on Karner Blue Butterfly (Lycaeides melissa samuelis) Habitat

The Karner blue butterfly (KBB) (Lycaeides melissa samuelis), a federally listed species, has historically been a component of barren and savannah ecosystems in the northeastern and midwestern United States. In New York, it now occurs primarily on managed, early‐successional sites such as power line corridors (utility rights‐of‐way) from Albany to Glens Falls. Blue lupine (Lupinus perennis), the exclusive larval food source of the KBB, has been suggested to be the most limiting factor for the butterfly within the eastern portion of its range. Power line corridor management maintains early‐successional habitats by suppressing the regrowth of woody species, creating potentially important habitat for the conservation and restoration of the KBB complex. This research compared the effects of several vegetation‐clearing methods on Blue lupine populations and associated communities of nectar species for KBB over an 8‐year period. Methods evaluated differed in intensity (annual, 4‐, or 8‐year intervals) and type (herbicide or mechanical). Blue lupine and plant community responses did not significantly differ among the treatment types applied to the power line corridors. However, Blue lupine cover, clump size, and density of stems per clump increased following the application of treatments in general. The number and cover of nectar species, total herbaceous cover, and species richness also responded positively to treatment overall. The percentage of non‐native species temporarily increased following treatment but declined to near pre‐treatment levels again as woody cover increased. Temporal changes in plant community composition were not related to management methods. The management tools assessed, mowing and herbicide application, will accomplish the goal of halting or reversing succession, maintaining critical habitat for the KBB in a landscape that provides little other suitable habitat.     

Purification of thiosulfate sulfurtransferase by selective immobilization on blue agarose

A new method for isolating crystalline bovine liver thiosulfate sulfurtransferase has been developed which relies on the selective binding of the enzyme to agarose-immobilized Cibacron Blue F3GA. This preparation has the advantages of simplicity, reproducibility, and rapidity. It is also suggested that the introduction of a binding and elution step will materially aid previously published procedures.     

Laplace's law and the alveolus: a misconception of anatomy and a misapplication of physics

Both the anatomy and the mechanics of inflation of the alveoli, as presented in most textbooks of physiology, have been misunderstood and misrepresented. The typical representation of the acinus as a "bunch of grapes" bears no resemblance to its real anatomy; the alveoli are not independent little balloons. Because of the prevalence of this misconception, Laplace's law, as it applies to spheres, has been invoked as a mechanical model for the forces of alveolar inflation and as an explanation for the necessity of pulmonary surfactant in the alveolus. Alveoli are prismatic or polygonal in shape, i.e., their walls are flat, and Laplace law considerations in their inflation apply only to the very small curved region in the fluid where these walls intersect. Alveoli do not readily collapse into one another because they are suspended in a matrix of connective tissue "cables" and share common, often perforated walls, so there can be no pressure differential across them. Surfactant has important functions along planar surfaces of the alveolar wall and in mitigating the forces that tend to close the small airways. Laplace's law as it applies to cylinders is an important feature of the mechanics of airway collapse, but the law as it applies to spheres is not relevant to the individual alveolus.