Biochemically marked lymphocytoid lines: establishment of Lesch-Nyhan cells

Two lymphocytoid cell lines have been established from a patient with the Lesch-Nyhan syndrome. The cells are deficient in hypoxanthine-guanine phosphoribosyltransferase, as demonstrated by their failure to incorporate [H(3)]hypoxanthine and by their inability to grow in medium in which they were nutritionally dependent upon exogenous hypoxanthine. This represents the first establishment of presumptively permanent human lymphocytoid cell lines that are deficient in a specific enzyme.     

Reproducible cloning assays for in vitro growth of canine hematopoietic progenitor cells and their potential applications in investigative hematotoxicity

A variety of in vitro cloning assays have been used for studying hematopoiesis in mice and human beings. However, these techniques have had limited use in dogs, a species used extensively as a model for hematopoietic research, particularly hematotoxicity. We have adopted cloning assays for in vitro growth of canine colony-forming unit-erythroid (CFU-E) and colony-forming unit-granulocyte/macrophage (CFU-GM) progenitor cells, using modified microplasma clot and soft agar culture systems, respectively. Marrow mononuclear cells separated by density-gradient centrifugation were added to the aforementioned culture systems. Erythroid colonies were stimulated with sheep plasma erythropoietin and incubated at 37 C in 5% CO2 for 2 days. The CFU-E colonies were fixed with 5% glutaraldehyde, stained with benzidine, counted, and expressed as a mean of 8 replicates. The CFU-GM colonies were stimulated with pooled serum from endotoxin-treated dogs and incubated for 8 days at 37 C in 10% CO2. Using an inverted microscope, the CFU-GM colonies were counted and expressed as a mean of 6 replicates. The number of colonies was proportional to the plated cell concentrations. The addition of 10% autologous serum to CFU-GM cultures increased the number of colonies by 80 to 100%, but markedly reduced the size and number of CFU-E colonies. The marrow cloning capacity among dogs of comparable age was similar, and little variation was noticed when bone marrow cells from the same dogs were cultured repeatedly over a period of 3 to 4 months. We concluded that these cloning assays are fast, reliable, and reproducible and that they allow quantitative determination of canine hematopoietic progenitor cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

Submergence of the Connecticut Coast

Radiocarbon-dated samples show that the Connecticut coast has submerged about 9 feet in the last 3000 years and about 33 feet in the last 7000 years. The rate of submergence is similar to rates reported from other coasts. The finding strengthens the hypothesis that a worldwide postglacial rise of sea level is the cause.     

GABA catabolism: localization of succinic semialdehyde dehydrogenase in brain motor and sensory nuclei

The localization of brain succinic semialdehyde dehydrogenase, a specific gamma-aminobutyric acid degradative enzyme, could potentially yield valuable information concerning the function of the enzyme. Application of a new histochemical technique for this enzyme has revealed characteristic patterns of neuronal staining that are contsistent within embryologically and functionally similar nuclei of the brainstem of the rat.     

Matrix modification to improve the recovery of MMHg from clear water using distillation

The use of distillation from aqueous solution dramatically improves the recovery of methyl mercury (MMHg) from complex water samples, as compared to solvent extraction techniques. However, low and irreproducible spike recoveries are often observed when distillation is applied to very clear water samples (i.e., precipitation, double deionized water (DDW), etc.) or those containing high chloride concentrations. Based upon the observation that recoveries and reproducibility are higher for waters containing strong complexing compounds, such as humic matter, we investigated the potential of matrix modification to improve the reproducibility of MMHg by distillation from more difficult matrices. At sample acid concentrations of less than 0.1M hydrochloric acid (HCl), we found that the addition of complexers such as ammonium pyrrolidine dithiocarbamate (APDC), and aqueous humic substances improved distillation spike recoveries and reproducibility. The use of complexers was also found to suppress the co-distillation of divalent mercury (Hg(II), which, in highly contaminated samples can be an interferant with the MMHg determination. At higher HCl concentrations, irreproducibility was dominated by the interference caused by chloride co-distilled with the water. In these cases, neither the addition of complexing agents, or buffers to bind the free hydrogen ions (H⁺) in solution were effective in improving results.     

Electron microscopic study of the unique features and structural-morphologic relationship of canine bone marrow

Electron microscopy revealed several unique features in canine bone marrow, compared with that of other species. The marrow was fatty and extensively trabeculated and was enclosed by a complete layer of endosteal bone-lining cells. Branched reticular cells were closely associated with each other and, occasionally, covered part of the sinus wall as an adventitial layer. The extent of adventitial coverage varied markedly and was less extensive, compared with that of other species. On average, only 23% of the sinus wall was covered by adventitial layer, in contrast to 65% reported in laboratory animals. Unilaminar sinuses, with no adventitial coverage, accounted for greater than 38% of all sinuses. Quantitative analysis indicated that 60% of the latter sinuses contained apertures, as opposed to 35% of sinuses with adventitial coverage (P less than 0.05). Moreover, the number of apertures in unilaminar sinuses was significantly (P less than 0.009) greater than that in multilaminar sinuses. Apertures were observed every 59 microns in unilaminar sinuses, in contrast to every 109 microns in multilaminar sinuses. Approximately 75% of the apertures were occupied by cells in transit, and only 25% were free of cells. Macrophages were distributed throughout the marrow and were closely associated with all blood cell lines. Occasionally, cells that entered the lumen were not fully mature. Erythroblasts were seen migrating across the wall and within the lumen of sinuses. The less-extensive adventitial coverage in canine bone marrow might indicate that the rate of cell delivery from the marrow into the circulation was relatively high in this species.(ABSTRACT TRUNCATED AT 250 WORDS)