Corticotroph,somatotroph and mammotroph cell kinetics in the postnatal infant female rat

The aim of this work was to detect if hypothalamic-pituitary maturation was accompanied by significant proliferation changes in differentiated pituitary cell pools. For this purpose, pituitary corticotroph (Ct), mammotroph (Mt) and somatotroph (St) proliferation activities were scanned in intact female rats during the postnatal (P) period (1–35 postnatal days). The techniques of tritiated thymidine labelling, immunostaining and autoradiography were combined to visualize DNA synthesis of hormone containing cells. Immunoreactive cell densities were measured using image analysis, and double labelled cells were counted. Corticotroph proliferation activity increased significantly on day P12, followed by an increase in the Ct proportion on days P13–14. This is the first observation of a spontaneous change of corticotroph proliferation at the end of the stress nonresponsive period. The mammotroph density and proliferation rate increased gradually during postnatal maturation, until the Mt pool overran other cell types of the female hypophysis on day 35. The somatotroph pool was the most numerous until day P20; the proliferation rate remained constant while St proportions increased reaching a plateau between days P13 and 20, then decreased to the adult level. Each cell type examined showed a characteristic, individual density and proliferation pattern.     

Behavioral reproductive isolation among sympatric strains of Brachionus plicatilis Müller 1786: insights into the status of this taxonomic species

We present results on cross-mating experiments using Brachionus plicatilis strains collected in three ponds of a coastal marsh (Torreblanca Marsh, Castellón, Spain). These strains were known to differ widely both in morphology and allozyme patterns from a previous study, where they were grouped into three genetically different clonal groups. Although some of the strains co-occurred in the same pond and sexual periods overlapped, no gene flow was found among them. Our first objective was to determine whether behavioral reproductive isolation was responsible for the absence of interbreeding. A second objective was to explore the relationship between sexual isolation and genetic divergence. We performed two experiments. In Experiment 1, we tested five strains from different clonal groups; in Experiment 2, we added a strain from a congeneric species, and strains from different ponds. We recorded male mating behavior in all possible male-female strain pairings. Our data show that males of a strain tend to mate with females of the same strain or genetically similar strains, regardless of the pond they come from. We also found a high positive correlation between isolation distance and genetic distance. These results support the view that mating behavior acts as an important isolating barrier giving cohesion to clonal groups, and structuring populations of this rotifer, and that Brachionus plicatilis is a taxon composed of more than one biological species.     

Conservation of highly repetitive DNA in cetaceans

It is controversial whether odontocetes (toothed whales) and mysticetes (whalebone whales) have a common ancestry. Cetacean karyological uniformity, which is unique among mammalian orders, suggests a monophyletic origin; however, several anatomical authorities have maintained that odontocetes and mysticetes are diphyletic. We investigated the issue using Southern blot hybridization. Two labelled restriction fragment probes from the DNA of the sei whale (a mysticete) were hybridized to restricted DNA of cetacean species representing all extant families except the Eschrichtiidae, the gray whales. The probes hybridized to specific restriction fragments in all odontocete and mysticete materials. Hybridizations showed preservation of hybridization homologies and a striking conservation of the length of highly repeated DNA sequences. The results are compatible with a common ancestry of odontocetes and mysticetes.     

Polarization and fluorescent microscopy —Simple methods for demonstrating pulmonary surfactant lipids

Summary Birefringence and fluorochrome lipid staining with benzpyrene are demonstrated as simple morphological methods to reveal the presence or absence of surfactant lipids in human newborn lung tissue. Lack of lipid birefringence proves to be an associated finding in the lungs of premature infants with hyaline membrane disease, indicating the possible pathogenetical importance of surfactant deficiency.     

The fluctuations in the level of endogenous growth regulators in seeds ofAcer pseudoplatanus L. in the course of stratification

In the course of cold stratification ofAcer pseudoplatanus L. fruits a statistically significant alternation occurs in their seeds of a rise and fall in the level of endogenous growth regulators. In the initial weeks the inhibitory effect slightly declines, or, on the contrary, the stimulatory effect slightly increases; in the middle phase of stratification a marked increase in inhibitions and reduction of stimulations appears, and towards the end of stratification the stimulatory effect of isolated substances in the individual biotests rises again, or their inhibition effect is decreased. No direct dependence was found between the decrease of the degree of dormancy and the drop of inhibitor, or increase of promotor levels. However, a certain analogy was observed between the time course of fluctuations in the level of growth regulators and the germination energy of the seeds investigated. An enhancement of the growth activity of the substances isolated (e.g. those of a gibberellin-like nature) in the last weeks of stratification can already be considered as the result of the release of fruits from dormancy.     

Molecular properties of the red cell calcium pump: II. Effects of calmodulin,proteolytic digestion and drugs on the calcium-induced membrane phosphorylation by ATP in inside-out red cell membrane vesicles

In inside-out human red cell membrane vesicles /IOV/, in the absence of Mg²⁺, the only calcium-induced labelling by γ³²P-ATP occurs in a 140–150 000 molecular weight protein fraction, representing the hydroxylamine-sensitive phosphorylated intermediate /EP/ of the calcium pump. In the presence of Mg²⁺ calcium-induced phosphorylation is accelerated but several other membrane proteins are also phosphorylated through protein kinase action forming hydroxylamine-insensitive bonds. Addition of calmodulin accelerates EP formation both in the absence and presence of Mg²⁺.Treatment of the membrane with SH-group reagents significantly reduces EP formation. Mild trypsin digestion of IOVs, stimulating active calcium transport, eliminates calmodulin action and decreases the steady-state level of EP. In trypsin-digested IOVs the molecular weight of the ³²P-labelled EP is shifted to lower values /110–120 000/ We suggest that trypsin digestion cleaves off a 20–40 000 molecular weight calmodulin-binding regulatory subunit of the calcium pump molecule.     

A study of the role of histidine side-chains at the active centre of amylolytic enzymes

The role of histidine side-chains in reactions catalysed by porcine-pancreatic alpha-amylase, sweet-potato beta-amylase, and Aspergillus niger glucamylase has been studied by using diethyl pyrocarbonate, a specific protein reagent. Changes in the activity, binding affinity, and apparent kinetic parameters due to ethoxycarbonylation have been determined. For pancreas alpha-amylase, four of the eight histidine groups, for sweet-potato beta-amylase, six of the seven histidine groups, and for A. niger glucamylase, four of the six histidine groups were shown to be ethoxycarbonylated. Ethoxycarbonylation occurred as an apparent first-order reaction, with rate constants in the range 3.6–4.9 x 10^?2min^?1. Ethoxycarbonylation of the histidine group at the active centre rapidly inactivated alpha-amylase, whereas the other three groups are not located in the active centre, although activity and substrate binding are only slightly affected by their modification. For beta-amylase and glucamylase, only slight or no change in activity could be detected on ethoxycarbonylation, whereas significant changes were observed in the binding affinity.     

The effect of cytokinins on nitrate reductase activity

Cytokinins in addition to nitrate induce nitrate reductase activity (NRA) in some plants. Effects of cytokinins onNRA was investigated in stem pith parenchyma of kale, intact wheat and barley seedlings and isolated cucumber cotyledons. The most profound effect onNRA was found in barley and wheat seedlings.NRA in seedlings sprayed with 100 μM 6-benzylaminopurine (BAP) for three subsequent days was increased in leaves and decreased in roots. These changes were further enhanced in seedlings grown in nutrient solution lacking nitrate:NRA in wheat and barley leaves was increased by 57% and 202%, respectively, in plants supplied with nitrate theNRA increase was not significant: in wheat and barley leaves by 22% and 9%, respectively. Similar effect of BAP and kinetin was found in kale stem parenchyma and cucumber cotyledons. The cytokinin kinetin or BAP alone increasedNRA about twice in kale and three times in cucumber. Addition of nitrate to the medium enhanced the effect of kinetin in kale discs, but the two effects were not additive. Additive effect of nitrate and BAP onNRA was found in cucumber cotyledons in light. In general NRA was more affected by cytokinins in intact seedlings of wheat and barley as compared to explanted tissue of kale and cucumber, and lack of nitrogen made their effect more expressive.     

Monoclonal antibodies to rabbit liver cytochrome P-448

Cytochrome P-448 (mol wt 55,000 Daltons) from rabbit liver was purified to a specific content of 16.6 nmol/mg. Mice were immunised with this preparation, their spleens removed and dissociated lymphocytes hybridised with myeloma cells. Four monoclonal antibodies against cytochrome P-448 were raised and partially characterised. All four antibodies interacted with cytochrome P-448 in intact microsomal fractions and selectively immunoadsorbed cytochrome P-448 from solubilised microsomal preparations. One of the antibodies inhibited benzo[a] pyrene hydroxylase activity in a reconstituted system, one had no effect on activity and two increased activity. The possible applications of such antibodies are discussed.     

Conservation and chromosomal localization of DNA satellites in balenopterid whales

DNA satellites were isolated from three balenopterid species, viz. the minke, sei, and fin whales. In each of them at least two DNA satellites were recognizable with buoyant densities in neutral CsCl of =1.702/1.703 and =1.710/1.711, respectively. cRNAs from each satellite group were used for filter and in situ hybridisations. Homo- and heterologous DNA-cRNA hybrids within each satellite group yielded virtually identical melting curve profiles showing conservation of at least a considerable part of the DNA satellite sequences. There was no evident sequence homology between the =1.702/1.703 and the =1.710/1.711 satellites by filter hybridisation. — The in situ hybridisation showed that in each species the =1.702/1.703 satellite was located in centromeric-paracentromeric C-bands in a few pairs, whereas the =1.710/1.711 satellite was located in terminal C-bands throughout the karyotypes. — The data on the whale DNA satellites indicate that the quantitative evolution of the satellite DNA sequences preceded species divergence of the balenopterids and that the satellite sequences have remained relatively unaltered since the divergence took place. The function of satellite DNA is considered to imply the introduction of both chromosomal and genic polymorphisms and thus being of great importance in speciation. Based upon these concepts a model is postulated for the function of satellite DNA. According to this model at meiotic pairing euchromatin-heterochromatin overlapping between homologous chromosomes is considered to be of a general occurrence. This overlapping is presumed to be accentuated by the size heteromorphism frequently observed between homologous heterochromatic segments (C-bands). In the region of such euchromatin-heterochromatin overlapping, crossing-over would be excluded. The overlapping is suggested to be rectified progressively in the chromosome arms, leaving unaffected crossing-over distant to the euchromatin-heterochromatin junctions. The consequence of this will be that genes in the proximity of the junctions are collectively inherited and selected, whereas genes distant to the heterochromatin will be independently assorted and selected.