The effects of cyclophosphamide on the gonadotrophic cells of the normal rat

The effects of the administration of cyclophosphamide to male rats at doses of 400 mg/m2 for a period of 5 days and of 200 mg/m2 over two five-day cycles interrupted by a 21-day break reveal differences in the ultrastructural morphology of gonadotrophic cells between treated animals and normal controls. The ultrastructural data obtained coincide with the results obtained from the measurement of serum FSH and LH levels, indicating hypofunction. A significant fall was also recorded in blood testosterone levels. The two types of treatment did not give rise to morphological or functional differences.     

Alterations of G-protein coupling function in phosphoinositide signaling pathways of cells transformed by ras and other membrane-associated and cytoplasmic oncogenes.

We showed previously that transformation by cytoplasmic and membrane-associated oncogenes including ras results in uncoupling between surface stimulation by platelet-derived growth factor, bombesin, and serum and activation of intracellular phospholipase C (PLC); this uncoupling does not involve alterations at the receptor or effector enzyme levels (T. Alonso, R. O. Morgan, J. C. Marvizon, H. Zarbl, and E. Santos, Proc. Natl. Acad. Sci. USA 85:4271-4275, 1988). In this study, we stimulated normal and oncogene-transformed NIH 3T3 cells with fluoroaluminate (AIF4-), thus directly activating PLC-associated G protein(s) and bypassing the receptor step. A1F4(-)-elicited PLC responses were significantly impaired in transformed cells when compared with those in their normal counterparts, suggesting that the uncoupling of PLC is the result, at least in part, of functional impairment at the G-protein level. Transformation by ras oncogenes has also been reported to result in enhanced PLC response to bradykinin resulting from increased receptor numbers (G. Parries, R. Hoebel, and E. Racker, Proc. Natl. Acad. Sci. USA 84:2648-2652, 1987; J. Downward, J. de Gunzburg, R. Riehl, and R. Weinberg, Proc. Natl. Acad. Sci. USA 85:5774-5778, 1988). We demonstrate here that transformation by other membrane-associated and cytoplasmic oncogenes also results in increased responsiveness to bradykinin ("supercoupling") and enhanced receptor numbers. However, there is no direct correlation between the number of receptors and the enhancement in responsiveness, suggesting that other factors besides receptor number are also involved in the enhanced responses. We propose that a common effect of transformation by cytoplasmic and membrane-associated oncogenes is functional alteration of coupling G proteins and that a similar modification of different kinds of G proteins may account for the pleiotropic alterations of signal transduction (uncoupling and supercoupling) observed.     

Molecular analysis of the replication region of the conjugative Streptococcus agalactiae plasmid pIP501 in Bacillus subtilis. Comparison with plasmids pAM beta 1 and pSM19035.

The large conjugative plasmid pIP501 was originally isolated from Streptococcus agalactiae. To study the molecular basis of pIP501 replication we determined the nucleotide sequence of a 2.2 kb DNA segment which is essential and sufficient for autonomous replication of pIP501 derived plasmids, in Bacillus subtilis cells. This region can be divided into two functionally discrete segments: a 496 bp region (oriR) that acts as an origin of replication, and a 1488 bp segment coding for an essential replication protein (RepR). The RepR protein, which has a molecular mass of 57.4 kDa, could complement in trans a thermosensitive replicon bearing the pIP501 origin. Chimeric Rep proteins and replicons were obtained by domain swapping between rep genes of closely related streptococcal plasmids belonging to the inc18 group (pIP501, pAM beta 1 and pSM19035). The chimeras were functional in B. subtilis.     

Cross-reaction of snRNA and an Alu I-like sequence from rat with DNAs from different eucaryotic species.

Sequence homologies to rat U1-snRNA and U2-snRNA were investigated in DNA from 23 eucaryotic species (3 lower eucaryotes, 4 plants, and 16 animals) using dot hybridization at various stringency conditions. Cross-hybridization among very distantly related species in e.g. plants-insects or mold-vertebrates is not the rule; there are, however, examples for stronger homologies like Rattus-Dictyostelium. Furthermore, DNA from all 23 species was analysed for sequence homologies with the repetitive DNA sequence B1 (an Alu I family equivalent) from rat. We observed a wide range of homologies covering some plants and insects, up to vertebrates. Hybridization at increasing stringency conditions revealed species with higher degree of homology to the rat B1 sequence: maize, chicken, and hamster.     

Observations on the diagenetic behavior of arsenic in a deep coastal sediment

Vertical profiles of total dissolved arsenic, manganese and iron, pH, Eh and rates of sulfate reduction were determined in a freshly-collected box core from a 335m depth station in the Laurentian Trough. The relationships observed between the profiles were further examined in the laboratory by measuring these same parameters with time in surficial sediment slurries as the Eh decreased in response to biological activity or chemical alteration.Both field and laboratory observations have shown that arsenic is released predominantly as As(III) into reducing sediment porewaters. This occurs after the dissolution of manganese oxides and at the same time as the dissolution of iron oxyhydroxides and the onset of sulfate reduction. Laboratory experiments indicated that sulfate reduction and the production of sulfide ions are not solely responsible for the release of arsenic to the porewaters, although this process is necessary to create and maintain a highly reducing environment conducive to rapid iron dissolution.The diagenesis of arsenic in Laurentain Trough sediments involves the simultaneous release of arsenic and iron at a subsurface depth, followed by its removal from porewaters by precipitation and adsorption reactions after migration by diffusion along concentration gradients. A qualitative model is presented to describe the behavior of arsenic in coastal marine sediments.Present address: Department of Geological Sciences, McGill University, 3450 UniversityStreet, Montreal, Quebec H3A 2A7, Canada     

Antagonism betweenYersinia intermedia andYersinia enterocolitica in water

One strain of Yersinia enterocolitica and one strain of Y. intermedia were grown in peptone water at 25 or 37 degrees C, or in ground water at 15 degrees C. Similar growth rates were observed when these strains were cultivated separately in the same media and at the same temperature. Mixed cultures at 37 degrees C displayed equivalent growth rates. In contrast, mixed cultures incubated at 15 or 25 degrees C were regularly unfavourable to Y. enterocolitica, whereas they did not modify the growth of Y. intermedia. A bacteriophage active on Y. enterocolitica and not on Y. intermedia was characterized from the filtrate of mixed cultures at low temperatures. This phage produced by the lysogenic Y. intermedia strain might be a potential factor responsible for the inhibition of Y. enterocolitica, since no additional antibacterial factor or nutritional competition between Y. intermedia and Y. enterocolitica were found in the mixed cultures.     

Genetic analysis ofHairy-wing mutations

Summary We studied the genetic bases of threeHairy-wing (Hw ¹,Hw ^Ua ,Hw ^49c ) mutations mapping in the region of theachaete-scute complex (AS-C). Analysis of X-ray-induced revertants ofHw ¹ andHw ^49c uncoveredachaete andscute mutant phenotypes respectively. This indicates that theHw mutant phenotypes result from an excess of function of these genes of theachaete-scute complex (AS-C). The phenotypes of the differentHws show allelic specificity in the pattern of extrachaetes. In addition to these mutations, certain inversions and internal duplications of the AS-C also produce aHw-variegated phenotype, probably due to variegation or decompensation of the genes of the AS-C. The expressivity of the differentHws (mutation or variegation) is modulated by the number of doses of the AS-C present in the genome. A similar dose-dependent modulation is exerted by the transregulatory geneshairy andextramacrochaetae. We discuss these results on the basis of a regulation model of the expression of the AS-C.     

Extraction of histones H2A, H3 and H4 from yeast nuclei. Measurement of the extent of yeast histone acetylation following one-dimensional gel electrophoresis

Yeast histones H2A, H3 and H4 were specifically extracted from purified nuclei using a 2% NaCl/75% ethanol solution. The extraction resulted in the complete removal of H2A, H3 and H4 from the nuclear pellet, as monitored by SDS-polyacrylamide gel electrophoresis of the protein. The relative absence of nonhistone proteins from this histone subset simplifies the determination of the extent of histone modification in yeast. Levels of H4 acetylation were measured directly on Coomassie blue-stained Triton acid-urea gels and the levels verified by gel fluorography of the [3H]acetate-labeled histone.     

Solid-state geometry and conformation of linear,diastereoisomeric oligoprolines

We have carried out a systematic analysis of the solid-state conformational preferences of a number of linear homo-oligoprolines (to the tetramer) by ir absorption and x-ray diffraction. The peptides present different chiral sequences (tacticities), various types (urethane and amide) of N-protecting groups, and free and blocked C-termini (which imply different capabilities of forming H-bonds). The following conclusions can be drawn: (i) values for the geometry of the prolyl residue and the peptide bond in the cis and in the trans conformations are proposed; (ii) in general the conformational angles φ and ψ in the linear homo-oligoprolines have values appropriate for the polyproline II structure (conformation F); (iii) the pyrrolidine ring shows various types of puckering with no apparent relation to the backbone conformation; (iv) Pro-Pro peptide bonds generally take the trans conformation, the few cases of cis conformation being formed by Pro residues of different chirality; (v) the single H-bond donor — OH, when present, is always bonded to H-acceptors, which can be either the urethane or the amide or the peptide carbonyl but never the carbonyl group of the — COOH moiety.