Isolation of an acyl-CoA carboxylase from the tapeworm spirometra mansonoides

An acyl-CoA carboxylase, which catalyzes the carboxylation of acetylpropionyl-, and butyryl-CoA, has been isolated from the tapeworm $\text{Spirometra}\text{mansonoides}$. The enzyme has an absolute requirement for ATP, Mg²⁺, and HCO₃${}^{\mathrm{?}}$ and, in addition, requires K⁺ for full catalytic activity. The enzyme has been purified 50-fold by a combination of calcium phosphate gel adsorption, ion-exchange column chromatography, and gel filtration. In its substrate specificity, K⁺ requirement, molecular size, and antigenic behavior, the tapeworm enzyme is similar to the acyl-CoA carboxylase of another helminth— the free-living nematode $\text{Turbatrix}\text{aceti}$.     

Plant uptake of bicarbonate as measured with the11C isotope

Summary ¹¹C which is cyclotron produced by¹⁴N(P, )¹¹C(half-life 20.1M) was use as a tracer of bicarbonate to determine its movements from a nutrient solution through roots to stems and leaves of bush bean plants (Phaseolus vulgaris L. var. Improved Tendergreen). The short time involved and the high solution pH minimized the need for use of the Hederson Hasselbach equation for activity correction. Quantities of¹¹C did move into roots, stems and leaves with a sharp decreasing gradient (root/stem=14.5, stems/leaves=11.7) More¹¹C moved into plants with KHCO₃ than with NaHCO₃. The (NH₄)₂SO₄ enhanced¹¹C uptake and KNO₃ than with competition indicated possibility of some uptake of HCO ₃ ^– . In an experiment withGalenia pubescens (Eckl. and Zeyh.) Druce, the¹¹C was more readily moved to stems and leaves than in bush bean indicating substantial uptake of HCO ₃ ^– .     

A multinational Andean genetic and health program. VIII. Lung function changes with migration between altitudes.

Studies of lung function in high altitude populations have suggested the influence of hypoxic environment on the development of this characteristic independent of confounding variables such as ethnicity and habitual exercise. However, often the effect of altitude on vital capacity is greater in children than adults, suggesting that more than developmental adaptation is operative. Also selective migration could account for the similarity of migrants and permanent residents at a destination altitude. To explore these problems we studied the lung function (FVC, FEV1, PFR) of 377 individuals who had migrated between altitudes in northern Chile. Migrant measurements were adjusted to those of permanent residents of appropriate age, sex and height at the altitudes of origin and destination. The measurements were then related to ethnicity (Spanish-Aymara ancestry), occupation and permanence, the latter combining information on both age at migration to and length of stay at a destination altitude. Upward migration was associated with increased chest depth, FVC and FEV1, but not height or other chest measurements. Downward migration had no significant effect. The flow-dependent test PFR was so sensitive to observer variability and occupation that it was difficult to establish its relationship to permanence. Unlike the body measurements, lung function measurements (especially PFR) tended to deviate from permanent controls at the origin altitude in a direction suggestive of selective migration, nor was permanence itself independent of ethnicity and occupation. Because of these difficulties the question of developmental adaptation in lung function may not be answerable in cross-sectional studies like the present and previous efforts, but rather in longitudinal investigations in which the control is the individual him/herself.     

A multivariate analysis of fatness and relative fat patterning.

Skinfold measurements (triceps, subscapular, suprailiac and medial calf) in four samples (376 boys, 352 girs, 338 men and 380 women from rural Colombia) were subjected to principal components analysis to identify components of obesity and relative fat patterning. Three components emerged which were similar in the four samples: a first component of fatness explaining 70-80% of the variance and two fat pattern components each explaining 10-15% of the variance: trunk-extremity and upper-lower body. Fatness and the trunk-extremity pattern components changed with age in children (7-12 years), but none of the components changed with age in adults (25-60+). The fatter tended to be more patterned in both age groups. Canonical correlation analysis revealed that socioeconomic status was more related to fatness than to patterning. With the exception of brothers, all first degree relatives (sib, parent-off-spring) and spouses were correlated in fatness. Some of the correlations between relatives--usually sibs, but not spouses--were also significant for the pattern components, suggesting a genetic basis for the known stability of this characteristic (Garn, '55a). Principal components analysis is a useful multivariate alternative for quantitative studies of anthropometric patterning.     

Effect of endo-beta-galactosidase on intact human erythrocytes.

Endo-beta-galactosidase, a glycosidase that hydrolyzes Gal beta 1-4 GlcNAc linkages in glycoconjugates, has been used to probe the plasma membrane of human erythrocytes. Coomassie blue staining of stroma components separated by sodium dodecyl sulfate-acrylamide gel electrophoresis indicates that treatment of red cells with endo-beta-galactosidase converts Protein 3, the anion transporter of the erythrocyte, to a more compact staining band. No other components detected by Coomassie staining are affected. Following labeling of red cells with galactose oxidase + NaB3H4, 45 to 50% of the [3H]galactose residues can be released by endo-beta-galactosidase. In contrast, only 5% of the label incorporated by treatment with periodate + NaB3H4, can be removed. [3H]Galactose residues are released from three components: Protein 3, Band 4.5, and the megaloglycolipids. The susceptibility of these components to endo-beta-galactosidase, together with the high content of Gal and GlcNAc present in Protein 3 and the megaloglycolipids, suggests that the erythrocyte membrane contains several components with N-acetyllactosamine repeating units, a structure commonly found in connective tissue glycoconjugates.     

Comparisons of diisopropylfluorophosphate derivatives of chymotrypsin and chymotrypsinogen by phosphorus-31 nuclear magnetic resonance

The nature of the differences in the active sites of α-chymotrypsin and chymotrypsinogen has been investigated by phosphorus-31 NMR studies of their diisopropylfluorophosphate derivatives. The phosphorus-31 resonance of the modified zymogen occurs 2 ppm upfield from that for the enzyme. An even greater separation is seen between diisopropylphosphoryl-neo-chymotrypsinogen and -α-chymotrypsin. A plausible interpretation of the chemical shift differences is based on the known structures for α-chymotrypsin, chymotrypsinogen and diisopropylphosphoryl-trypsin.     

The presence and possible function of methylmalonyl CoA mutase and propionyl CoA carboxylase in Spirometra mansonoides.

Both spargana and adult forms of Spirometra mansonoides were shown to accumulate lactate, succinate, acetate, and propionate upon in vitro incubation. Adults differed markedly from the spargana in that quantitatively the most significant products of the former were acetate and propionate while the latter formed primarily acetate and lactate. The adults accumulated approximately 32 times more propionate than the spargana per gram of tissue. In accord with this propionate formation, propionyl CoA carboxylase and methylmalonyl CoA mutase have been found to be present in both stages of the parasite. As might be predicted, however, the activities of the carboxylase and mutase were 100-fold and 10-fold higher, respectively, in the adults as compared to the larvae. A possible physiological relationship between propionate formation and energy generation is suggested. Accordingly, inorganic 32P was incorporated into ATP upon incubation of methylmalonyl CoA with a homogenate obtained from adult S. mansonoides. Since methylmalonyl CoA mutase requires vitamin B12 coenzyme, a relationship between vitamin B12 content and propionate formation in helminths is suggested.     

Iron regulatory protein 1 as a sensor of reactive oxygen species

Iron regulatory proteins (IRP1 and 2) function as translational regulators that coordinate the cellular iron metabolism of eukaryotes by binding to the mRNA of target genes such as the transferrin receptor or ferritin. In addition to IRP2, IRP1 serves as sensor of reactive oxygen species (ROS). As iron and oxygen are essential but potentially toxic constituents of most organisms, ROS-mediated modulation of IRP1 activity may be an important regulatory element in dissecting iron homeostasis and oxidative stress. The responses of IRP1 towards reactive oxygen species are compartment-specific and rather complex: H2O2 activates IRP1 via a signaling cascade that leads to upregulation of the transferrin receptor and cellular iron accumulation. Contrary, superoxide inactivates IRP1 by a direct chemical attack being limited to the intracellular compartment. In particular, activation of IRP1 by H2O2 has established a new regulatory link between inflammation and iron metabolism with new clinical implications. This mechanism seems to contribute to the anemia of chronic disease and inflammation-mediated iron accumulation in tissues. In addition, the cytotoxic side effects of redox-cycling anticancer drugs such as doxorubicin may involve H2O2-mediated IRP1 activation. These molecular insights open up new therapeutic strategies for the clinical management of chronic inflammation and drug-mediated cardiotoxicity.     

53BP1 cooperates with p53 and functions as a haploinsufficient tumor suppressor in mice

p53 binding protein 1 (53BP1) is a putative DNA damage sensor that accumulates at sites of double-strand breaks (DSBs) in a manner dependent on histone H2AX. Here we show that the loss of one or both copies of 53BP1 greatly accelerates lymphomagenesis in a p53-null background, suggesting that 53BP1 and p53 cooperate in tumor suppression. A subset of 53BP1-/- p53-/- lymphomas, like those in H2AX-/- p53-/- mice, were diploid and harbored clonal translocations involving antigen receptor loci, indicating misrepair of DSBs during V(D)J recombination as one cause of oncogenic transformation. Loss of a single 53BP1 allele compromised genomic stability and DSB repair, which could explain the susceptibility of 53BP1+/- mice to tumorigenesis. In addition to structural aberrations, there were high rates of chromosomal missegregation and accumulation of aneuploid cells in 53BP1-/- p53+/+ and 53BP1-/- p53-/- tumors as well as in primary 53BP1-/- splenocytes. We conclude that 53BP1 functions as a dosage-dependent caretaker that promotes genomic stability by a mechanism that preserves chromosome structure and number.