Halobacterial megaplasmids are negatively supercoiled

Several covalently closed circular halobacterial megaplasmids (up to more than 500 kb) from different strains of Halolerax mediterranei, have been resolved by orthogonal-field alternating gel electro-phoresis (OFAGE). These molecules seem to be negatively supercoiled in vivo, as deduced from the effect of intercalating agents affecting their topology and, therefore, their electrophoretic mobility. It has also been demonstrated that the topolsomerase II Inhibitor novobiocin affects the native topological state of halobacterial megaplasmids impeding their migration in OFAGE under standard conditions for resolution of large supercoiled molecules.     

Continuous-flow NMR culture system for mammalian cells

A continuous-flow NMR culture system for mammalian cells has been developed on which ³¹P-NMR experiments under complete and strictly physiologic conditions have been performed. Observations on the response of the cellular metabolism to stresses such as starvation, low temperature and changes in environmental pH monitored by ³¹P-NMR are reported. The response of the intracellular pH relative to the external pH of the growth medium is studied. We find that under the experimental conditions used there exists a ΔpH varying between less than 0.2 and more than 0.6 pH units. These results are compatible with those obtained using other techniques.     

The economics of monocropping and intercropping by smallholders: The case of coconuts in Indonesia

This article contains a comparison of the profits of cultivating modern and traditional varieties of coconuts as a monocrop and as an intercrop, in ideal and in average growing conditions, with good and with average management. We carry out the analysis from a private (financial) and from a national (economic) perspective. The results show that intercropping generates more income than monocropping. In the conclusion we discuss why development organizations and Third World countries encourage monocropping.     

Purification and characterization of the high molecular weight microtubule associated proteins from neonatal rat brain

The changes in the levels of microtubule-associated proteins (MAPs) during advanced embryonic stages, neonatal and adult organisms reflect the importance of these cytoskeletal proteins in relation to the morphogenesis of the central nervous system. MAP-1B is found in prenatal brains and it appears to have the highests levels in neonatal rat brains, being a developmentally-regulated protein. In this research, a fast procedure to isolate MAP-1B, as well as MAP-2 and MAP-3 from neonatal rat brains was designed, based on the differential capacity of poly L-aspartic acid to release MAPs during temperature-dependent cycles of microtubule assembly in the absence of taxol. The high molecular weight MAP-1B was recovered in the warm supernatants after microtubular protein polymerization in the presence of low concentrations of polyaspartic acid. Instead, MAP-2 and a 180 kDa protein with characteristics of MAP-3 remained associated to the polymer after the assembly. Further purification of MAP-1B was attained after phosphocellulose chromatography. Isolation of MAP-2 isoforms together with MAP-3 was achieved on the basis of their selective interactions with calmodulin-agarose affinity columns. In addition, MAP-2 and MAP-3 were also purified on the basis of their capacities to interact with the tubulin peptide -II (422–434) derivatized on an Affigel matrix. However, MAP-1B did not interact with the -II tubulin fragment, but it showed interaction with the Affigel-conjugated -I (431–444) tubulin peptide. The different MAPs componentes were characterized by western blots using specific monoclonal antibodies. A salient feature of neonatal rat brain MAP-3 was its interactions with site-directed antibodies that recognize binding epitopes on the repetitive sequences of tau and MAP-2. However, these site-specific antibodies did not interact with MAP-1B from the neonatal rat brain tissue.Abbreviations PAA poly (L-aspartic acid) - HMW-MAPs high molecular weight microtubule associated proteins     

Coprecipitating IgG asymmetric antibodies: A possible role for Fab glycosylation,and speculations on their formation and functions in disease

IgG asymmetric antibodies are synthesized by the same cellular clones as the symmetric ones but appear in the immune response in different proportions. The evidence suggests that they are caused by asymmetric glycosylation on some IgG molecules in the Fab region. The cause of this is unknown but it could be speculated that there are cellular factors that induce glycosyl transferases or cause the molecule to be more accessible to glycosylation. The production of asymmetric antibodies can be modified by the physical status (soluble or particulate) of the antigen used as immunogen by the number and frequency of stimulation, and by physiological factors such as the ones secreted by the placenta and by lymphocytes that express progesterone receptors in response to hormone. An increase of these antibodies can be beneficial or harmful to the host, depending on the situation in which they act and the character of self or non-self of the antigens recognized. Editors note—Many of the ideas proposed in this article are very speculative, but it was thought appropriate to publish it in order to stimulate further discussion of the subject. It is an interesting role for Fab glycosylation that is proposed by Professor Margni. The ideas discussed are not necessarily those held by the Editorial Board or the reviewers, who felt that the evidence for many of the deductions made was very limited. It was also emphasized by the reviewers that the author's case would be substantially improved if more corroborative evidence was available from other groups. The Editors would welcome any comments on the subject for publication in future issues of the journal.     

Formaldehyde-induced appearance of septate junctions between digestive vacuoles

Tissue biopsies from (1) some chronic inflammatory diseases, (2) a necrotic tumoral process, (3) normal human lymphatic ganglia, and (4) two congenital diseases of the adrenal cortex were selected for study. A block from each biopsy was fixed in glutaraldehyde-paraformaldehyde; a second block was fixed in 10% formaldehyde. In all cases septate junctions between digestive vacuoles did occur in phagocytic cells and some adrenal cortex cells fixed in formaldehyde. These junctions were similar to those reported recently for malakoplakia phagocytes. Consistently, they were not found to attach organelles other than lysosomes derivatives. Both phagocytes and adrenal cortex cells in the material fixed in glutaraldehyde-paraformaldehyde did not display adhesive specializations between digestive vacuoles. This suggests that the septate junctions described herein are artifactuous structures induced by formaldehyde. There is, however, a certain degree of specificity of cells having the capability of developing these septate junctions. It is assumed that the coating material of digestive organelles in phogocytes and some other cells would be responsible for both cell specificity and organelle specificity of the formaldehyde-induced septate junctions.     

INFLORESCENCE SIZE AND FRUIT DISTRIBUTION AMONG INDIVIDUALS IN THREE ORCHID SPECIES

Fruit distribution among individuals in populations of three orchid species was analyzed. Fruit set was low in the three species. The distribution of fruits in the non-autogamous species was skewed to the right, with high variance: mean ratios, and a moderate to high proportion of fruitless plants. Larger inflorescence size did not necessarily result in an increase in percent fruit set but tended to increase the probability to produce at least one fruit. Pollinator-mediated selection on inflorescence size through fruit production may be limited by the low overall level of visitation and the resulting uncertainty of pollination at the individual level.