On the Trail of Atmospheric Mutagens and Carcinogens: A Combined Chemical/Microbiological Approach

Major ecological problems of our polluted troposphere includeairborne toxic chemicals, acid rain and photochemical smog,all three of which are now recognized as being closely relatedchemical phenomena. We also recognize that inorder to developcost-effective strategies for their control, which protect publichealth and the environment, there must be close scientific interactionsbetween chemists and biological scientists. For example, ofrapidly emerging importance is the development of risk assessmentevaluations for specific aspects of each of these problem areas.In preparing such assessments, chemists must define the "exposure,"and biological scientists the "effects." In this paper, I discuss an example of how such close interactionsproved indispensible in our search for atmospheric mutagensand carcinogens. Thus, an integrated chemical/ microbiologicalprocedure for the isolation and identificationof particulatechemical mutagens in respirable diesel soot and ambient particlesis described. Emphasis is placed on our use of the short-term,Ames Salmonella typhimurium bacterial mutagenicity test as arapid, and relatively inexpensive, means of following the biologicalactivities of these environmental mutagens through the chemicalsteps of their separation, isolation and identification fromhighly complex environmental samples. Possible mechanisms offormation of these particulate mutagens are discussed. Theyinclude the reactions of polycyclic aromatic hydrocarbons presenton the surfaces of combustion-generated particles with gaseousco-pollutants such as nitrogen dioxide plus nitric acid, andozone. In discussing this research on a societally "relevant" problem,we illustrate the importance of "Science as a Way of Knowing."We further suggest that this integrated approach to scientificproblem solving by chemical and biological scientists mightserve as an example of a discussion topic on human ecology forundergraduate courses in the natural sciences.     

Instability kinetics of trp operon plasmid ColEl-trp in recombinant Escherichia coli MV 12[pVH5] and MV 12trp R [pVH5

In studying plasmid instability in recombinant microorganisms, Escherichia coli MV 12[p VH5] and MV 12 trpR[p VH5] harboring trp operon were used as experimental model systems. The host with the trp repression system was partially derepressed by 3-beta-indoleacrylic acid. The results from kinetic analysis of plasmid instability showed that the stability of p VH5 and the growth rate of MV 12[p VH5] decreased rapidly in presence of 3-beta-indoleacrylic acid at a concentration higher than 10 mug/mL, but beyond 30 g/mL no significant change was observed. This suggests that Trp(-) variants from MV 12[p VH5] could be produced from the host cells at a different frequency depending on the physiological condition. In another system, MV 12 trpR[p VH5] which was constructed by conjugation of E. coli MV 12[p VH5] with E. coli CSH61, the plasmid stability was much lower and the frequency of Trp(-) cell production was about 10 times higher as compared with the MV 12[p VH5] when treated with 3-beta-indoleacrylic acid. A kinetic model representing the plasmid instability was derived, and a fairly good agreement with the experimental results was found. The fraction of plasmid-free cell (or negative variant) shows a different time course profile depending on the segregation coefficient (a, production rate of negative variants from positive cells during one generation), growth ratio (G, the ratio of growth rates of negative variants to positive cells), and other parameters. The negative variant fraction continues to increase ("run away" type) or it approaches to a finite value ("settling" type) depending on the relative values of a and G.     

ESTIMATES OF CARRY-OVER EFFECTS IN TWO-PRODUCT HOME USAGE CONSUMER TESTS

In-house use consumer test data from four studies dealing with three pairs of household products and a pair of antiperspirant products were examined for significant carry-over (product usage order) effects, which would confound the analysis of treatment (product) effects. In each study, two products were compared using a two-period crossover design. One hundred twenty panelists participated in each study. A forced choice preference scale or a 9-point hedonic scale was used to obtain responses from various sensory attributes. In all studies, the estimates of carry-over effects were not significant at the 5% level. Transformation of hedonic scale data into preference dichotomy also gave estimates of carry-over effects which were not significant at the 5% level, but led to a loss of test sensitivity for detecting treatment differences. The authors recommend that all comparative crossover design studies in sensory evaluation be monitored for carry-over effects and that statistically determined sample size should be used to reduce the possibility of obtaining significant carry-over effects.     

Polymorphism and stability of histone gene clusters in Drosophila melanogaster cultured cells

In a previous communication (Saigo, K., Millstein, L. and Thomas, C.A., Jr. (1981) Cold Spring Harbor Symp. Quant. Biol. 45, 815–827), the overall structure of histone genes of Schneider line 2 cells was shown to extensively differ from that of Oregon-R embryo from which the cell line was established, and it was speculated that the histone genes might be reshuffled extensively during either the periods of the establishment, or maintenance of cell lines, or both. To establish the validity of this notion the structure of histone genes was examined in Drosophila melanogaster cultured cells. The overall organization of histone gene clusters was found to be stably maintained in both the periods for the establishment and maintenance of cultured cells, indicating that the previous assumption is inadequate. Instead of an extensive rearrangement, minor structural changes were found to occasionally occur probably by simple base substitutions and/or, deletion or insertion of very short DNA pieces. It was also shown that the extensive variation in structures of histone genes in cultured cells such as Schneider line 2 are attributable to polymorphism on the level of individual flies.     

Ontogeny of the Behavior of Strombus maculatus (Gastropoda: Strombidae)

The ontogeny of strombid behavior was studied by observing thebehavior of Strombus maculatus veligers collected from the planktonand reared past metamorphosis to adults, and by observing juvenilestrombids collected in the field. Complete adult modal actionpatterns (MAP's) associated with locomotion, feeding, and rightingof overturned shells are performed by S. maculatus juvenilesimmediately after metamorphosis. There are changes in the frequencyof the use of certain MAP's which are associated with variationsin shell shape and size. The unique strombid escape response to molluscivorous gastropods(Conns spp.) is not released until juvenile S. maculatus arethree weeks past metamorphosis and two millimeters in shelllength. At that stage, the complete response is released uponthe first encounter with a predator. Experience with a predatordoes not seem to lower the age or size criteria. During ontogeny there is a trend toward an increasing complexityof behavior which is paralleled by an increasing complexityof neural structure and general morphology. There are majorsteps in the ontogeny of strombid behavior which probably coincidewith neural and morphological stages.     

Malarial Parasites of the "Jesu Cristo" Lizard Basiliscus basiliscus (Iguanidae) in Panama

SYNOPSIS. The iguanid lizard Basiliscus basiliscus in Panama is parasitized by Plasmodium basilisci and P. achiotense sp. nov. P. basilisci in this host is characterized by schizonts containing 4–14 merozoites, with schizonts parasitizing proerythrocytes containing more merozoites than those in erythrocytes. Asexual parasites lack cytoplasmic projections, while mature gametocytes are round or oval with regular margins.
P. achiotense is characterized by the combination of prominently pigmented, large schizonts containing 36–56 merozoites and oval or round gametocytes which are about 1/3 larger than those of P. basilisci.
EE-schizonts of P. basilisci were observed commonly in thrombocytes and occasionally in lymphocytes, and appeared early in experimental infections induced by blood inoculation.     

A New Saurian Malarial Parasite Plasmodium balli from Panama

SYNOPSIS. Plasmodium balli sp. nov. is described from Anolis lionotus and A. poecilopus of central Panama.
Large, elongate gametocytes and segmenters containing up to 100 merozoites are produced by P. balli. Proerythrocytes and normoblasts are more commonly parasitized than erythrocytes. Pigment is uncommon, but when present consists of a minute dot. Hypertrophy, distortion and lysis of host cell nuclei may result from parasitization of immature blood cells by gametocytes, while enucleated host cells are common.     

The Control of Color in Mammals

SYNOPSIS. Recent advances in the biology of mammalian pigmentationare reviewed.Particular attention is given to emerging informationon the integration of pigmentary events occurring at differentlevels of biological organization within mammalian skin andhair. The structural and functional significance of keratinocytesand melanocytes as components of mammalian epidermal melaninunits is viewed from this perspective. New evidence on the natureof genetic, developmental, endocrine, and radiation influenceson performance of melanocytes and the establishment of pigmentarypatterns is summarized.     

Differentiating Type i and Type ii Alveolar Cells in Rat Lung by OsO4NaI Staining

The fixing-staining mixture consisted of 1 part of 2% aqueous OsO₄ and 3 parts of 3% Nal in distilled water. Fresh lungs were cut into 2 mm slices and immersed in this solution for 24 hr at room temperature. Controls were fixed in buffered OsO₄ alone. Selective staining of type II alveolar cells was shown by the OsO_t-NaI mixture but was absent in the controls. No additional staining of the sections was required, and the selectivity was readily observable in either paraffin or Araldite sections by light microscopy and in Araldite sections by electron microscopy     

Saurian Malarial Parasites in Eastern Panama

SYNOPSIS Plasmodium gonatodi sp. nov. is described from Gonatodes albogularis fuscus of eastern Panama. It is characterized by elongate gametocytes and polymorphic schizonts containing 12-46 nuclei when apparently mature. Both proerythrocytes and erythrocytes are commonly parasitized, host cells are hypertrophied and distorted, and their nuclei are displaced. Prematuration sexual stages may be irregularly shaped and larger than mature gametocytes.
Plasmodium diploglossi Aragão and Neiva , 1909 is reported from Mabuya mabouya in eastern Panama, and Plasmodium morulum sp. nov. is described from this host. P. morulum usually parasitizes immature erythrocytes, and is characterized by lenticular or oval to round gametocytes, and schizonts with 14-40 nuclei usually arranged in a globular mass. Host cells are slightly hypertrophied and distorted, and their nuclei are usually displaced. Inoculation of infected blood into clean hosts produces numerous schizonts in white cells as well as in the erythrocyte series.
Pigment in both P. gonatodi and P. morulum , if present, consists of a few minute dark dots which do not meet the polarized light test for hemozoin.