In vitro stimulation of alkaline phosphatase activity in immature embryonic chick pelvic cartilage by adenosine

Cyclic AMP content in embryonic chick pelvic cartilage increases significantly as the embryo ages from 8 to 10 d. This in ovo elevation in cyclic AMP content precedes maximal cartilage alkaline phosphatase activity by some 24 h. We studied whether this temporal relationship may be causally related, using an in vitro organ culture. Incubation of pelvic cartilage from 9- and 10-d embryos in medium containing monobutyryl cyclic AMP (BtcAMP) resulted in significant increases in alkaline phosphatase activity (220 and 66 percent, respectively) as compared to that of cartilages incubated in medium alone. This stimulation was both concentration- and time-dependent with maximal response at 0.5 mM BtcAMP and 4-h incubation, respectively. Similar incubations of cartilage in medium containing 1-methyl-3-isobutyl xanthine (MIX), 0.25 mM, also resulted in increased alkaline phosphatase activity (114 percent). However, pelvic cartilage from 11-d embryos incubated in medium containing BtcAMP or MIX showed no increase in alkaline phosphatase activity. We postulated that developmental age was the factor responsible for this difference in response and that immature cartilage (that with little or no alkaline phosphatase activity) would respond to BtcAMP whereas mature cartilage (that with significant alkaline phosphatase activity) would not. This was tested by incubating end sections of 11-d cartilage, which have little alkaline phosphatase activity, and center sections, which have significantly alkaline phosphatase activity, with both BtcAMP and MIX. Alkaline phosphatase activity in end sections (immature cartilage) was stimulated by BtcAMP and MIX, whereas it was not stimulated in the center sections. Actinomycin D and cycloheximide inhibited BtcAMP and MIX stimulation of alkaline phosphatase activity. Thus, the in vitro data suggest that cyclic AMP is a mediator for the stimulation of alkaline phosphatase activity in embryonic cartilage.     

Binding of aldolase to actin-containing filaments. Evidence of interaction with the regulatory proteins of skeletal muscle.

The interactions of aldolase with regulatory proteins of rabbit skeletal muscle were investigated by moving-boundary electrophoresis. A salt-dependent interaction of troponin, tropomyosin and the tropomyosin-troponin complex with aldolase was detected, the tropomyosin-troponin complex displaying a greater affinity for the enzyme than did either regulatory protein alone. The results indicate that aldolase possesses multiple binding sites (three or more) for these muscle proteins. Quantitative studies of the binding of aldolase to actin-containing filaments showed the interaction to be influenced markedly by the presence of these muscle regulatory proteins on the filaments. In imidazole/HCl buffer, I 0.088, pH 6.8, aldolase binds to F-actin with an affinity constant of 2 x 10(5) M-1 and a stoicheiometry of one tetrameric aldolase molecule per 14 monomeric actin units. Use of F-actin-tropomyosin as adsorbent results in a doubling of the stoicheiometry without significant change in the intrinsic association constant. With F-actin-tropomyosin-troponin a lower binding constant (6 x 10(4) M-1) but even greater stoicheiometry (4:14 actin units) are observed. The presence of Ca2+ (0.1 mM) decreases this stoicheiometry to 3:14 without affecting significantly the magnitude of the intrinsic binding constant.     

On the optimal harvesting of persistent age-structured populations

Summary This paper discusses optimal harvesting policies for age-structured populations harvested with effort independent of age.     

Reduction of physical assignments to a standard lod table: Chromosome 1

Summary A method for reducing physical assignments of markers to a lod score table is described, and these tables are added to those from family data to provide a consistent genetic map of chromosome 1. Various chiasma maps are considered, and the results suggest that terminalization of chiasmata does occur. Chiasmata appear to be significantly less localized in oocytes than spermatocytes, and crossing-over in the vicinity of the centromere is more common in females than in males.This work was supported by Grants GM 23021, GM 24941, and NF 1-475 from the U.S. National Institutes of Health and National Foundation, PGL Paper No. 228     

Age,area, and acheiropody

Summary Analysis of the geographic distribution of acheiropody suggests spread from São Paulo along the São Francisco valley at a rate consistent with current parent-offspring distribution and 20 generations of gene flow.PGL No. 233. This work was supported by Grants GM 17173 from the U.S. National Institutes of Health and Biologicas 79/0482 from São Paulo Research Foundation (FAPESP).     

Biological and cultural determinants of immunoglobulin levels in a Brazilian population with Chagas' disease

Summary Under a general model the genetic heritability is 0.33 for IgA, 0.34 for IgG and 0.12 for IgM, in a Brazilian population with Chagas' disease. Cultural heritability is much smaller. The analyses suggest that, with respect to family resemblance for immunoglobulins, there is no discrepancy between this sample and those from healthy populations reanalyzed recently (Barbosa et al. 1981).This work was supported by Grants 79/0482 FAPESP (Brazil), 77/2222.1392 CNPq (Brazil) and GM 17173 (Hawaii) with collaboration of the Fundação Instituto Osvaldo Cruz (Bambui, MG). Partly supported by NIH Grants GM 24941 and GM 28719.     

Oncofetal antigen-I (OFA-I) relative antigenicity on melanoma cells

Summary The oncofetal antigen-I (OFA-I) has been defined as an immunogenic antigen that is expressed on human cancer cells and is cross reactive with fetal brain tissue. Quantitative variations in the expression of OFA-I among different cultured melanoma cell lines were determined by absorption techniques based on functionally monospecific anti-OFA-I serum. Allo-antibodies were removed by absorption with lymphoblasts autologous to an OFA-I-positive target cell. Functional monospecificity toward OFA-I was confirmed by complete absorption with a specimen of fetal brain but not by liver from the same fetus.Of 14 melanoma cell lines tested, two did not express OFA-I, whereas 12 expressed the antigen to varying degrees. Five of the cell lines were highly antigenic, and serum absorbed with 5×10⁵ of any of these cell lines could reduce the anti-OFA-I titer (1 : 96) at least four-fold. OFA-I was detected on biopsied melanomas autologous to the antigenic cultured cells. The ability to select highly antigenic cell lines could be useful in further attempts to characterize OFA-I and to monitor tumor immunity in vitro. Antigenic cell lines may improve the response of patients treated in trials of immunotherapy.     

Inhibition of normal allogeneic lymphocyte mitogenesis by a factor released from human tumor cells in culture

Summary Human tumor and normal cell lines in culture were examined for the release of factors capable of inhibiting lymphocyte blastogenesis. Supernatants from tumor cell cultures of melanoma, carcinoma (lung, colon, breast), sarcoma, and normal fibroblasts inhibited normal lymphocyte response to PHA. Only supernatants from the tumor cell lines C1 (colon carcinoma), 734B (breast carcinoma), and 231 (breast carcinoma) were found to inhibit both PHA and ConA responses significantly. The two breast carcinoma cell lines, 734B and 231, which also were capable of inhibiting lymphocyte responses to PPD and alloantigens, were investigated further. The inhibition of lymphocyte blastogenesis caused by the supernatant of these two cell lines could not be overcome by the addition of added mitogen. Further experiments showed that the inhibition was not due to nutrient deficiencies and the supernatants were not directly toxic to the lymphocyte cultures as judged by trypan blue exclusion.