A single species,two basic chromosomal numbers: case of Lygeum spartum (Poaceae)

Abstract

The existence of two chromosome numbers (2n?=?16 and 2n?=?40) in Lygeum spartum is confirmed in the Algerian steppe populations of Oran region. Chromosome counts were established for 11 Algerian populations and three supplementary populations from Italy, Spain and Greece. The karyotypes were characterized by chromosome markers obtained using fluorescence in situ hybridization and fluorochrome bandings. The organization of 5S and 18S-5.8S-26S (35S) rDNA was studied in both cytotypes. Six signals of 35S and 2 signals of 5S were observed in 2n?=?16 population, while 10 signals of 35S and 4 signals of 5S were detected in the population with 2n?=?40. All 35S loci were also strongly marked by chromomycin, but negatively stained by Hoechst, which indicates the presence of GC rich DNA in rDNA regions. The B chromosomes were found in both cytotypes, bearing a 35S locus in 2n?=?16 population. Genome size, determined by cytometry of 10 populations, ranged from 9.27?pg for 2n?=?16 to 26.63?pg for 2n?=?40 populations. The sequencing of plastid and nuclear DNA markers did not reveal major differences among 2n?=?16 and 2n?=?40 populations. However, given the differences between two cytotypes and based on their morphological and cytogenetic characteristics, the 2n?=?16 cytotype merits novel taxonomic treatment.     

Effects of S-100 Proteins on Assembly of Brain Microtubule Proteins: Correlation Between Kinetic and Ultrastructural Data

Microtubules formed in vitro in the presence of S-100 proteins and micromolar Ca2+ concentrations are fewer in number and longer than those formed in the presence of Ca2+ alone. Moreover, microtubules growing after addition of microtubule fragments to a microtubule protein solution in the presence of S-100 are shorter than those growing in its absence. These data lend support to previous results of kinetic studies indicating that S-100 interferes with both the nucleation and the elongation of microtubules in vitro.     

Can one predict protein stability? An attempt to do so for residue 133 of T4 lysozyme using a combination of free energy derivatives,PROFEC, and free energy perturbation methods

Free energy derivatives, pictorial representation of free energy changes (PROFEC) and free energy perturbation methods were employed to suggest the modifications that may improve the stability of a mutant T4 lysozyme with a S-2-amino-3-cyclopentylpropanoic acid residue (Cpe) at position 133. The free energy derivatives and PROFEC methods were used to locate promising sites where modifications may be introduced. The effects of several candidate modifications on the enzyme's stability were analyzed by the free energy perturbation method. We found that this scheme is able to effectively suggest modifications that may increase the enzyme's stability. The modifications investigated are the introduction of a methyl, a tert-butyl or a trifluoromethyl group at the C_ϵ2 position and a cyclopropyl group between the C_δ2 and C_ϵ2 position on the cyclopentyl ring. The stereochemistry of the introduced groups (in the α or β configurations) was studied. Our calculations predict that the introduction of a methyl group in the α configuration or a cyclopropyl group in the β configuration will increase the stability of the enzyme; the introduction of the two groups in the other configurations and the other modifications will decrease the stability of the enzyme. The results indicate that packing interactions can strongly influence the stability of the enzyme. Proteins 32:438–458, 1998. © 1998 Wiley-Liss, Inc.     

Neuron-specific enolase and S-100 protein immunohistochemistry for defining the structure and topographical relationship of the different enteric nerve plexuses in the small intestine of the pig

Summary The morphological and topographical features of the intramural enteric nervous system in the small intestine of the pig has been studied on whole mounts by means of neuron-specific enolase (NSE) and S-100 protein immu-nohistochemistry. A clear visualization of the myenteric plexus allows the recognition of its characteristic morphology, including the thin tertiary plexus coursing within the smooth muscle layers. In the tela submucosa two ganglionated plexuses, each with its own specific characteristics, can clearly be demonstrated: (1) the plexus submucosus externus (Schabadasch) located near the inner surface of the circular muscle layer at the abluminal side of the submucosal vascular arcades, and (2) the plexus submucosus internus (Meissner) close to the outer surface of the lamina muscularis mucosae at the luminal side of the submucosal vascular arcades. Due to the possibility to trace clearly the perivascular plexuses of these vascular arcades by use of immunohistochemical techniques with antibodies to NSE and S-100 protein, the two submucosal nerve plexuses can be demonstrated with exceptional clarity. This is the first report of an investigation of the intramural nerve plexuses of the small intestine of the pig using the NSE and S-100 immunostaining methods, which is sufficiently detailed to substantiate the characteristic topography and structure of the two submucosal plexuses and their relation to the smooth muscle layers and perivascular plexuses. The level of NSE immunoreactivity for enteric neurons displays great variation, a substantial proportion of the type-II neurons appearing strongly stained. Although little is known of the specific function of these enzymes, proposals are discussed.     

Effects of fluorescamine labeling on mitochondrial ATPase activity.

Fluorescamine labeling of rat liver mitochondria enhances the ATPase activity. It reached maximum stimulation when mitochondria were treated with 30–34 nmol fluorescamine per mg of mitochondrial protein. This stimulation is inhibited by N,N′-dicyclohexylcarbodiimide. The maximum stimulation caused by labeling is the same as that obtained from uncoupler with optimum concentration. The chemiosmotic potential (Δμ_H⁺) decreases as the labeling increased. However, Δμ_H⁺ is not abolished completely even when ATPase activity reaches a maximum. The results suggest that primary amino groups may be involved in controlling mitochondrial ATPase activity.     

Metabolic effects of some electrofluorimetric dyes

The effect of five electrofluorimetric dyes on mitochondrial metabolism was examined to determine their suitability for mitochondrial studies and other biological uses. The dyes merocyanine 540, 8-anilino-1-naphthalene sulfonic acid and bis(1,3-dibutyl barbituric acid-(5))-pentamethane oxonol were found to be inhibitors of the respiratory chain. However, the first two exerted their effect only at high concentrations. 3.3′-Dihexyl-2,2′-oxacarbocyanine was found to act as an uncoupler. 3,3′-Dipropyl-thiocarbocyanine inhibited β-hydroxybutyrate respiration while dissociating succinate supported respiration from the phosphorylation of ADP. Merocyanine 540 and 8-anilino-1-naphthalene sulfonic acid may be the best suited for studies of membrane potentials in mitochondria since their effect on metabolism is negligible.     

Structure and function of human fetal endocrine pancreas before and after cryopreservation

Human fetal pancreatic glands obtained from 31 consecutive prostaglandin-induced abortions were examined with respect to light microscopic structure and insulin content and release before and after cryopreservation. The crown-heel lengths of the fetuses ranged from 12 to 34 cm. Minced pancreatic fragments about 2 mm³ in size were cultured overnight in RPMI 1640 medium plus 10% fetal calf serum. The explants were incubated at 0 °C for 20 min in Hanks' solution containing 1 M Me₂SO and subsequently cooled at 0.3 °C/min to ?70 °C before rapid quenching in liquid nitrogen. After storage for 4–150 days at ?196 °C the pancreatic fragments were rapidly thawed and suspended in RPMI 1640 (10% calf serum) for another overnight culture.After cryopreservation there was some morphological deterioration of the fetal pancreas. Before cryopreservation 13 of the fetal glands responded with a significant insulin release to an acute glucose plus theophylline challenge, while after cryopreservation 16 glands responded.Although cryopreservation lowered the insulin response there was a strong statistical correlation between the response obtained before and after freezing (P < 0.001). No correlation could be demonstrated between the insulin response and crown-heel length either before or after freezing. There was no obvious effect of cryopreservation on the pancreatic insulin content which showed a significant correlation with the crown—heel length both before and after freezing.It is concluded that cryopreservation of human fetal endocrine pancreas preserves the viability of the B cells. These observations provide a basis for further exploration of the suitability of human fetal pancreas for clinical transplantation.