Permanent Stained Preparations of Synovial Fluid for Detection of Calcium Compounds Using Alizarin Red S

Permanent preparations of air dried synovial fluids were prepared by staining calcium compounds with alizarin red S stain; each slide was coverslipped with Permount. Variables studied were: (a) concentration of the solution of alizarin red S, (b) pH of staining solution, (c) time of incubation in staining solution and aqueous and ethanolic content of staining solution. The staining effect of each solution was tested on calcium pyrophosphate dihydrate, calcium oxalate, apatite and monosodium urate (MSU). Of all the solutions, best results were obtained with 0.25% alizarin red S in 50% ethanol at pH 7.0 for 30 min. With this solution, the calcium-containing compounds were well stained. MSU did not stain and still preserved negative birefringence on polarizaton. Fixation of smears with ethanol served a double purpose: It fixed the slides without dissolving or removing MSU or the calcium compounds, yet it did dissolve five corticosteroids commonly used for intra-articular injection which may interfere with interpretation of compensated polarized light microscopy of synovial fluids.     

Developmentally regulated lectin in dark versus white axolotl embryos

The white mutant of the Mexican axolotl, A. mexicanum, involves an ectodermal defect which prevents melanophore colonization. Endogenous lectins have been suggested to function in neural crest-derived melanophore adhesion in other animals. To determine if differences in endogenous lectins exist in dark and white axolotls during melanophore colonization, white and dark ectoderm and carcass tissues have been assayed for lectin activity at premigratory, early migratory, and late migratory neural crest stages. Lectin content (specific for D-glucosamine, N-acetyl-D-glucosamine and D-mannose) increases significantly during early migration only in dark ectoderm and white carcass tissues, whereas white ectoderm and dark carcass lectin activities remain close to premigration levels. Neural crest cells in these embryos are associated with regions of high lectin activity suggesting that the differences in endogenous lectins may be involved in establishment of the dark/white phenotype.     

MICROSPOROGENESIS IN PEANUT (ARACHIS HYPOGAEA)

The stage of pollen development at the time of anther culture is an important factor in the production of haploids. The objectives of the current study were to develop a staining procedure for peanut (Arachis hypogaea L., ssp. hypogaea) microspores, to describe and document the stages of microsporogenesis in peanut, and to confirm a previous report concerning correlations of peanut floral bud shape with stage of microspore development. A staining procedure using propionic carmine provided adequate staining of pollen mother cells, microspores, and pollen. Pollen mother cells and microspores could easily be differentiated by their size and cell wall structure. Plants grown in a controlled environment were found to have highly synchronized microspore development, both within an anther and among anthers contained in the same bud. In addition, floral bud shape was confirmed as a reliable indicator of anther stage in peanuts.     

Dopamine produces muscle contractions and modulates motoneuron-induced contractions inAplysia gill

1. Dopamine has been reported to exist in unusually large quantities in Aplysia gill. The physiological role of this neurotransmitter in this organ was examined. 2. The addition of dopamine to a gill perfusate results in the contractions of the lateral and medial external pinnule muscles, the circular and longitudinal muscles of the afferent vessel, and the circular muscles of the efferent vessel. 3. Dopamine-induced contractions persist after chemical synaptic transmission is eliminated in the gill. This suggests that excitatory dopamine receptors are present on gill smooth muscle fibers themselves. 4. Dopamine also potentiates the gill response to action potentials in single identified gill motoneurons. Evidence presented suggests that muscle contractions and modulation of motoneuron contractions are independent phenomena. 5. While modulation may in part be mediated by increases in excitatory junction potential (EJP) amplitude, in many cases large increases in muscle contractions occur while the enhancement of EJPs is disproportionately small. 6. Dopamine's ability to produce muscle contractions suggests that there may be dopaminergic motoneuron innervation of the gill. We suggest that dopamine's modulatory actions may be mediated via modification of excitation-contraction coupling in smooth muscle fibers.     

Precision of the all-glass impinger and the andersen microbial impactor for air sampling in solid-waste handling facilities.

A method was devised to determine the precision of the all-glass impinger and the Andersen six-stage microbial impactor over a wide range of aerosol concentrations like those found in facilities which process solid waste. Simultaneous samples were collected inside a municipal solid-waste recovery system, and the data were treated statistically to estimate the precision of each air-sampling device. All-glass impingers yielded colony counts which indicated a linear relationship between samplers over an observed aerosol concentration of 1.1 X 10(3) to 2.8 X 10(7) colony-forming units per m3 of air. Impactors also yielded colony counts which indicated a linear relationship over an observed aerosol concentration range of 3.9 X 10(3) to 1.9 X 10(5) colony-forming units per m3 of air. The coefficients of variation for the all-glass impinger and the six-stage impactor in an environment with a high and variable dust level were determined to be 0.38 and 0.23, respectively.     

Effects of heavy metals on structure,function, and metabolism of ciliated respiratory epithelium in vitro

Summary Tracheal explants were used to evaluate the relative ciliostatic and cytotoxic potential of heavy metal salts (cadmium chloride, chromium chloride, nickel chloride, and copper sulfate). Explants from hamster, rat, and guinea pig were all sensitive to the metals, though guinea pig explants showed the greatest difference between the untreated and metal treated tissues. Dosage levels were 50, 100, and 500μM, for 24 to 148 h. Cadmium caused the greatest degree of ciliostasis and cell necrosis. Copper was less toxic, and nickel and chromium caused marginal damage when tested at 100μM or lower. In each instance, damage became detectable at approximately 24 to 48 h and was nearly stabilized by 72 h. A significant loss of ciliary motion was always accompanied by a decrease in metabolic activity (dehydrogenase activity and ATP content). Transmission and scanning electron microscopy revealed a severely necrotic epithelium after exposure to cadmium, with only subtle morphological alterations after exposure to other metals. With all of the treatments there was no overt structural damage to cilia and little alteration in membranes of cells remaining in the epithelium. Some coagulation or vacuolization was noted in cadmium and copper treated explants but most cellular organelles did not display obvious damage. The most significant changes in the tracheal epithelium exposed to heavy metal salts in vitro were a loss of ciliary motion and a decrease in total ATP content. This study was supported in part by Grant HL 26880 from the National Heart, Lung and Blood Institute, Bethesda, MD.     

3H-noradrenaline metabolism in the isolated epididymal and prostatic portions of the rat vas deferens

The spontaneous ³H-noradrenaline efflux from the isolated epididymal and prostatic portions of the rat vas deferens, was investigated. The spontaneous tritium efflux was higher in the epididymal portion than in the prostatic one from normal animals. Such differences were abolished after castration. On the other hand, acetylsalicylic acid enhanced the total tritium efflux only in the epididymal portion, whereas phentolamine and phenoxybenzamine increased the total tritium efflux from the two portions of the rat vas deferens in both experimental groups. The prostatic portion released a similar amount of ³H normetanephrine than the apididymal end, whereas other metabolic fractions (3,4-dihydroxyphenylglycol; 3,4-dihydroxymandelic acid and O-methylated deaminated metabolites) were smaller in the prostatic portion in comparison with the epididymal portion from control vas deferens. The results presented in the isolated rat vas deferens suggest the existence of a prostaglandin as well as an alpha adrenoceptive modulation of the spontaneous total tritium efflux.     

Taxonomy of nonfermentative bacilli: The IIk-2 group

Twenty-two strains of the Center for Disease Control’s IIk-2 group of nonfermentative Gram-negative, weakly pigmented (yellow) bacteria were examined in respect to phenotypic features, DNA base composition, and both intra-and extra-group DNA-DNA hybridizations. Eighteen of the 22 strains appeared to be members of one taxon. This taxon is phenotypically not similar to two genera,Acinetobacter andMoraxella, which have similar base compositions. It does, however, have phenotypic features and base composition compatible with those of the genusFlavobacterium.