The distribution of fetal hemoglobin and the types of gamma chain in red cell fractions separated by gradient centrifugation from blood of patients with sickle cell anemia and other hemoglobinopathies

Isopycnic separations of red cells from cord bloods, and from patients with sickle cell anemia, different forms of HPFH, S-beta O-thalassemia, and a beta +-thalassemia homozygosity were made in order to evaluate the distribution of Hb F and the relative levels of G gamma and A gamma chains over the cell fractions. As expected, the cord blood data showed decreased levels of both Hb-F and G gamma chains in the top cell fractions since the beta leads to gamma and high G gamma: A gamma low G gamma: A gamma switches are operative around the time of birth. Complete cell fractionations were made on the blood of three SS patients with low G gamma values (40%) and three SS patients with high G gamma values (60%). The proportion of G gamma chain was constant in all cell fractions, while the Hb-F level was higher in cells with higher densities. The difference in the quantities of the three types of gamma chain in the fetal hemoglobins of two SS patients with an A gamma T heterozygosity, one having a low G gamma value and the other a high G gamma level, can be explained by assuming an alteration in a regulatory mechanism. Considerable variation both in the level of Hb F and in the percentage of G gamma chain was observed in two G gamma A gamma-HPFH heterozygotes with a relatively low G gamma percentage of 30%; an inverse relationship was present between the two parameters. Such a phenomenon was not evident for the G gamma A gamma-HPFH homozygote, and also did not exist in two additional G gamma A gamma-HPFH heterozygotes with an associated alpha-thalassemia-2 heterozygosity who had a similar amount of Hb F but with higher G gamma values of about 50%. The difference in G gamma values between these two categories of G gamma A gamma-HPFH could be due to a higher affinity of the G gamma chains over A gamma chains for a slightly decreased amount of alpha chains as in an alpha-thalassemia-2 heterozygosity. 2 heterozygosity. Although an increased synthesis of Hb-F with G gamma chains was again observed after in vitro incubation of reticulocytes with [35S]methionine none of the isolated cell fractions contained a Hb F with the alpha 2 G gamma 2 composition.     

How Escherichia coli sets different basal levels in SOS operons

The recA and sfiA genes of Escherichia coli are SOS operons regulated negatively by the LexA repressor. The steady state level of expression of recA is 10-fold higher than that of sfiA, as measured by means of recA::lac and sfiA::lac operon fusions. To study the molecular basis of this difference, we have compared the expression of these two operons in strains in which the concentration of LexA repressor was normal (lexA+), zero (spr amber mutation) or higher than normal (plasmid pJL45, carrying the lexA gene linked to the lac promoter). The results indicate (i) that the recA promoter is about 4 times stronger than the sfiA promoter (as measured in the spr strains), (ii) that neither operon has a physiologically significant level of lexA-independent expression (pJL45 strains), and (iii) that the recA operator has about 2.5 times lower affinity than the sfiA operator for LexA repressor (comparison of lex+ and spr strains). Considering our previous results that the sfiA operon (high operator affinity of LexA) is derepressed very rapidly after inducing treatments and that the recA operon (low operator affinity) is repressed very rapidly when induction is stopped, we conclude that differences in operator affinity do not affect inducibility but serve only to set the basal levels of the different SOS functions.     

Identification by Tn10 transposon mutagenesis of host factors involved in the biosynthesis of K99 fimbriae of Escherichia coli: Effect of LPS core mutations

Abstract Tn 10 transposon mutagenesis of Escherichia coli producing K99 fimbriae was carried out to identify host factors involved in regulation or biosynthesis of fimbriae. Two chromosomal mutants were obtained that showed a strongly reduced cell surface expression of K99 fimbriae upon colony blotting and ELISA. Analysis by inversed PCR and nucleotide sequencing showed that one mutant (EP14) contained the Tn 10 transposon in rfaQ , affecting theexpression of the rfaQGP gene cluster, whereas the other mutant (EP35) was affected in a, to date, unknown region of the genome. Immunoblotting analysis confirmed a Rd₁ type of LPS of mutant strain EP14. These findings for the first time indicated an effect of LPS core biosynthesis on the biogenesis of fimbriae at the cell surface. Preliminary experiments indicated that K99 major subunits, in contrast to K88 subunits, strongly bind LPS molecules.     

Selection of biting sites on a human host by shape Anopheles gambiae s.s., shape An. arabiensis and shape An. quadriannulatus

Adult females of the predatory mite Phytoseiulus persimilis Athias-Henriot are strongly attracted to infochemicals released by plants infested with their prey, the two-spotted spider mite (Tetranychus urticae Koch), thereby effectively locating their prey. However, we found a consistently lower degree of attraction to these infochemicals for a population of P. persimilis, which is called non-responding population. Here we demonstrate that this low degree of attraction is a contagious phenomenon and that it cannot be explained by differences in abiotic conditions, physiological state and experience of predators or by genetic differences between predator populations. Female predators exposed to dead conspecifics of the non-responding population and their products showed a lower degree of attraction to plant odours and a higher mortality than predators exposed to products of a living conspecific of the non-responding population. This was true 6–7 days after contact with dead conspecifics and their products whereas 2 days after contact no effects were detected. The present results are discussed in view of our hypothesis that the change in foraging behaviour as well as the high mortality rate are symptoms of a contagious disease affecting the non-responding population.     

Pulmonary extraction of dopamine in the conscious dog

Thus far non-respiratory pulmonary functions have mainly been studied in isolated lungs or in intact anaesthetized animals. Therefore, we studied pulmonary handling of vasoactive hormones in the conscious dog. Dopamine, adrenaline and noradrenaline (3 nmol/ml) were injected together with cardiogreen (4 mg/ml) into the caval vein during 5, 10 and 15 s in three separate experiments. Indicator-dilution curves were derived from samples originating from the pulmonary artery and the aorta. During a single passage of the lung dopamine was removed to the greatest extent, noradrenaline somewhat less, and adrenaline not at all, irrespective of the injection duration. In particular, the extraction of dopamine, which was not emphasized in literature, was a remarkable finding.