Insulin in Invertebrates and Cyclostomes

It seems increasingly clear that insulin is a hormone that doesnot occur exclusively in vertebrates. Several independent reportsnow exist giving evidence of insulin production in the digestivetissues of both deuterostomian and protostomian invertebrates.Cells with some light-microscopical and ultrastructural characteristicsof vertebrate B-cells have also been observed. Recently, evidencehas been obtained that insulin can act as a hypoglycemic hormone,promoting glycogen synthesis, also in a protostomian invertebrate,the gastropod mollusc, Strophocheilus oblongus. The endocrine pancreas of the cyclostomes occupies a key positionin the comparative endocrinology of the islet parenchyma andin the evolution of insulin production. It may represent anevolutionary link between the presumably gut-connected dispersedinsulin-producing cells of deuterostomian invertebrates andthe pancreatic islets of gnathostomian vertebrates. This hypothesiswas supported by the fact that cells with light-microscopicaland ultrastructural similarities to the islet B-cells were observedin the bile duct mucosa of the hagfish, Myxine glutinosa. However,immunofluorescent studies with antisera against human insulinand C-peptide did not show any immunoreactive material outsidethe B-cells of the endocrine pancreas. Particular attentionwas paid to elucidate the biological significance of the largecystic cavities that are so typical for the cyclostomian isletparenchyma. The working hypothesis that they may contain storedinsulin, proinsulin (or even "proto-proinsulin") was not supportedby immunofluorescence, autoradiographic, or ultrastructuralinvestigations, nor by proinsulin assays. It is possible thathagfish islet B-cells contain zinc, despite the fact that theamino acid residue in B₁₀-position is aspartic acid insteadof histidine. The biosynthesis of hagfish insulin shows a patternsimilar to that in gnathostomes. Its rate is related to theambient temperature and at 11 C the conversion of proinsulinto insulin lasts several days.     

Life Cycle of Isospora canis Nemeséri, 1959 in the Dog*

The life cycle of I. canis Nemeséri, 1959 was studied in experimentally infected dogs. Freshly sporulated oocysts were ovoid and 34–40 × 28–32 μm. The endogenous stages were found directly beneath the epithelium of the distal portion of the small intestinal villi. Most of the endogenous stages were in the lower 1/3 of the small intestine, but occasionally they were found in other portions of the small intestine. Three asexual generations were present. First-generation schizonts were 16–38 × 11–23 μm and contained 4–24 merozoites; mature 1st-generation merozoites were 8–11 × 3–5 μm. First-generation schizogony lasted up to 7 days after inoculation. Second-generation schizonts were 12–18 × 8–13 μm and contained up to 12 merozoites which were 11–13 × 3–5 μm. Second-generation schizogony was present on postinoculation days 6 and 7. Third-generation schizonts were formed by nuclear division of 2nd-generation merozoites. Most 2nd-generation merozoites underwent nuclear division without leaving the parasitophorous vacuole of the 2nd-generation schizont. Mature 3rd-generation schizonts were 13–38 × 8–24 μm and contained 6–72 merozoites. Third-generation merozoites were 8–13 × 1–3 μm. Third-generation schizogony was present on days 6–8 after inoculation. Mature macrogametes were 22–29 × 14–23 μm. Mature microgametocytes were 20–38 × 14–26 μm. Gametes were present on postinoculation days 7–10. Oocysts were present in tissue sections on postinoculation days 8–10 and 12. The prepatent period was 9–11 days.     

Biochemical Cytology of Trichomonad Flagellates. II. Subcellular Distribution of Oxidoreductases and Hydrolases in Monocercomonas sp.*

A primitive trichomonad, Monocercomonas sp. (strain NS-1:PRR) from Natrix sipedon, was grown axenically in Diamond's medium. Activity of NADH oxidase, malate dehydrogenase, malate dehydrogenase (decarboxylating) and of the anaerobic enzymes, pyruvate synthase and hydrogenase as well as of several hydrolases was demonstrated in homogenates. The subcellular distribution of these activities was studied by means of analytical differential and isopycnic centrifugation of homogenates prepared in 0.25 M sucrose. NADH oxidase and malate dehydrogenase are in the nonsedimentable part of the cytoplasm. Malate dehydrogenase (decarboxylating), pyruvate synthase, and hydrogenase are associated with a large particle which equilibrates at density 1.22. In its properties, this particle corresponds to the microbody-like hydrogenosomes of Tritrichomonas foetus. The 5 hydrolases studied are associated with at least 2 different particle populations: a large particle population equilibrating at densities from 1.10 to 1.16 is the exclusive location of 3 enzymes (β-galactosidase, protease and β-N-acetylglucosaminidase), 2 of which have a pH optimum close to neutrality. These particles contain part of the acid phosphatase and β-glucuronidase. Another part of these 2 enzymes is associated with a separate population of smaller granules with equilibrium densities of 1.16 to 1.18. The 2 types of hydrolase-carrying particles are also biochemically very similar to their counterparts in T. foetus.     

Involvement of <Emphasis Type="Italic">H</Emphasis>-2 Locus in a Multigenically-determined Immune Response

IT has been known for a number of years that specific immune responses can be under genetic control. In some cases the immunological phenotype is controlled by a single Mendelian genetic factor^1–3, while in other cases a more complex mode of inheritance determines the animal's ability to mount a specific immune response^4–7. An intriguing feature of this line of investigation is that in many cases a gene controlling a specific immune response is genetically linked to a histocompatibility locus^3,8–13. We now report that a specific immune response which has been shown to be under complex genetic control⁷ can be explained by the segregation of two genetic loci, one of which is closely linked or identical to the H-2 locus.     

New Type of Colony-forming Cell located in the Pleural Cavity

A semi-solid agar culture system has been developed which supports the clonal growth of granulocytic and/or macrophage colonies from their specific progenitor cells^1,2. In the course of studies on the level of these progenitor cells in the peripheral blood of mice, whole blood was cultured in agar-medium. Cultures were prepared in 35 mm plastic Petri dishes and each culture contained 1 ml. of 0.3% agar in modified Eagle's medium. The media and general technique of agar culture have been described elsewhere³. In these experiments, 0.2 ml. of whole, unheparinized blood was taken directly from the axilla of anaesthetized 3 months old C57BL mice and added to 5 ml. of agar-medium, held liquid at 37° C. One ml. portions of this mixture were pipetted into four replicate culture dishes, allowed to gel and incubated for 7 days at 37° C in a fully humidified atmosphere of 10% CO₂ in air. Each culture contained 0.1 ml. of a 1:6 dilution of pooled sera from C57BL mice injected 3 h previously with 5 µg endotoxin to provide an adequate concentration of the specific colony-stimulating factor (CSF), required for granulocytic and macrophage colony formation.     

<Emphasis Type="Italic">E. coli</Emphasis> DNA Polymerase I and other Host Functions participate in T4 DNA Replication and Recombination

The recognition of bacterial functions involved in DNA metabolism of bacteriophage T4 might reveal interactions between different enzymes during DNA replication and recombination. To detect such functions we have studied the replication of complete and incomplete T4 chromosomes in various mutant strains of Escherichia coli that are defective in their own DNA metabolism. We found that several E. coli functions can substitute for phage functions in T4 replication and recombination and will discuss here the role of the E. coli pol A gene which codes for DNA polymerase I^1–4 and of the dna B and E genes^3,5.     

Four New Species of Eimeria (Protozoa: Eimeriidae) from the Pika Ochotona princeps from Alberta and O. pallasi from Kazakhstan1

SYNOPSIS. Eimeria worleyi n. sp. and Eimeria barretti n. sp. from the pika Ochotona princeps from Alberta and Eimeria pallasi n. sp. and Eimeria shubini n. sp. from the pika Ochotona pallasi from Central Kazakhstan are described. Oocysts of E. worleyi were 12–16 by 10–15 μ (mean 13.5 by 12.5 μ) and spherical to subspherical. E. barretti oocysts were ellipsoidal to slightly ovoid and 27–36 by 21–27 μ (mean 32.9 by 23.8 μ). Oocysts of E. pallasi were ellipsoidal or ovoid and 19–34 by 17–26 μ (mean 26.3 by 21.3 μ). E. shubini oocysts were spherical and 22 μ in diameter. E. kriygsmanni Yakimoff and Gousseff, 1938 of Svanbaev (1958) in O. pallasi, [non] E. kriigsmanni Yakimoff and Gousseff, 1938 in Mus musculus, is a synonym of E. pallasi. E. musculi Yakimoff and Gousseff, 1938 of Svanbaev (1958) in O. pallasi, [non] E. musculi Yakimoff and Gousseff, 1938 in Mus musculus, is a synonym of E. shubini.     

Isolation and Characterization of Antigenic Components of a New Heptavalent <Emphasis Type="Italic">Pseudomonas</Emphasis> Vaccine

ONE of the most serious opportunistic bacterial pathogens is Pseudomonas aeruginosa¹ and the inability of the common antimicrobial agents to combat such infections suggested the investigation of specific immunological prophylactic and therapeutic approaches. Therefore Fisher et al. began to develop a new serotype schema as a prerequisite for an immunizing preparation which would protect humans against the most prevalent strains of Pseudomonas. This communication describes the isolation, purification and preliminary characterization of seven new lipopolysaccharide antigens which were obtained from strains representative of each of the seven protective serotypes identified by Fisher et al.².