Human hypertensive placenta contains an increased amount of Na,K-ATPase with higher affinity for cardiac glycosides

Placentas of women suffering from pregnancy-induced hypertension (PIH) were found to contain a greater amount of Na,K-ATPase molecules, estimated from anthroyl ouabain binding, than normotensive individuals. Both the microsomal fraction of placental cells and purified Na,K-ATPase showed an increased affinity for the specific inhibitor ouabain which, in the case of the microsomes, bound with a dissociation constant of 0.9 nM as compared with 3.4 nM in the controls. Likewise, the dissociation constant of the ouabain complex with purified Na,K-ATPase was about 3.5 times lower in the hypertensive patients. The differences are apparently caused by a different microenvironment of the ouabain-binding site, as reflected in the quantum yield of bound anthroyl ouabain. If an endogenous digitalis-like factor is present in the body fluids to regulate Na,K-ATPase activity, the present results render its role quite plausible.     

The Presence of (+)-S-Adenosyl-l-Methionine in the Rat Brain and Its Lack of Effect on Phenylethanolamine N-Methyltransferase Activity

Abstract: (+)-S-Adenosyl- l -methionine [(+)-SAM] was isolated from rat brain and was quantified by HPLC followed by UV spectrophotometric measurements and by ¹H-NMR. Its estimated ratio in brain is 3% of total SAM. Because of its commercial unavailability, (+)-SAM was also prepared from chemically synthesized SAM by separation of the two diastereoisomers on a preparative reverse-phase Nucleosil C8 column. The (+) diastereoisomer thus obtained was then assayed in vitro both as an inhibitor and a substrate of phenylethanolamine N -methyltransferase. Enzymatic activity was measured by HPLC analysis. It was shown that (+)-SAM has no effect on phenylethanolamine N -methyltransferase activity; therefore, it is unlikely that (+)-SAM plays any possible role in regulation of adrenaline synthesis in the brain.     

Topoisomerase activity associated with SV40 large tumor antigen.

Purified preparations of simian virus 40 (SV40) large tumor antigen (LT) from three different sources, including LT expressed from a recombinant baculovirus, were found to relax negatively supercoiled cyclic DNA molecules, whether or not they contained SV40 sequences. Relaxation was stimulated by MgCl2 but not by ATP, and inhibited by camptothecin, suggesting the involvement of an enzymatic activity similar to that of topoisomerase I (topo I). However, the pH requirements for relaxation by respectively LT and topo I are different. Also, antibodies reacting with LT inhibited relaxation by preparations of LT but not topo I, whereas antibodies inhibiting relaxation by topo I had no effect on relaxation by LT. Reconstruction experiments suggested that both procedures used to purify LT, immunoaffinity chromatography and DEAE-Sepharose chromatography, separate topo I from LT. Finally, relaxing activity was found in over 40 preparations of LT, and in the few instances where activity could not be found, it probably had been lost during storage, rather than absent from the start. Whereas these results seem to exclude that the activity being detected is that of a contaminant of LT, they would be consistent with this activity being that of a stable topo-LT complex, or else intrinsic to LT itself.     

High prevalence of a point mutation in the porphobilinogen deaminase gene in Dutch patients with acute intermittent porphyria

Acute intermittent porphyria (AIP) is an autosomal dominant disease characterized by a deficiency of porphobilinogen deaminase (PBGD). Up to now 14 different mutations have been described. In an effort to investigate the molecular epidemiology of AIP we have undertaken a systematic study of different exons of the PBGD gene from a large number of unrelated patients. Here, exon 8 from 82 unrelated Dutch and French AIP patients was examined using single strand confirmation polymorphism analysis (SSCP) after polymerase chain reaction (PCR) amplification. A single base mutation, C to T, at position 346 of the sequence coding for PBGD was observed in 15 Dutch families but in only 1 French family. A simple PCR assay is described to facilitate the diagnosis of this common mutation at the DNA level.     

The IS4 family of insertion sequences: evidence for a conserved transposase motif

The eight IS 231 variants characterized so far (IS 231 A-F, V and W) display similar transposases with an overall 40% identity. Comparison with all the proka-ryotic transposable elements sequenced so far revealed that the IS231 transposases share two conserved regions with those of 35 other insertion sequences of wide origins. These insertion sequences, defining the IS4 family, have a common bipartite organization of their ends and are divided into two similarity groups. Interestingly, the transposase domains conserved within this family display similarities with the well known integrase domain shared by transposases of the IS3 and IS15 families, and integrases of retroelements. This domain is also found in IS30- related elements and Tn7 TnsB protein. Amino acid residues conserved throughout all these prokaryotic and eukaryotic mobile genetic elements define a major transposase/integrase motif, likely to play an important role in the transposition process.     

Phenotypic effects of overexpression of Agrobacterium rhizogenes T-DNA ORF13 in transgenic tobacco plants are mediated by diffusible factor(s)

Nicotiana tabacum cv. Xanthi transgenic plants expressing ORF13 of Agrobacterium rhizogenes 8196 T-DNA under the 35S RNA promoter from the cauliflower mosaic virus displayed developmental abnormalities. They were small, with short and variable internodal lengths, their root systems were poorly developed; leaves were small, asymmetric, rounded, wrinkled and dark green; flowers were short, and irregularly shaped. They exhibited reduced apical dominance and regularly produced offshoots at the base of the plant. This phenotype was also exhibited by offshoots of normal N. tabacum cv. Xanthi stock grafted with a transgenic scion indicating that expression of ORF13 influences plant development via diffusible factor(s).     

A rapid assay for chloroplast-encoded triazine resistance in higher plants

We have designed a simple and rapid assay for chloroplast-based triazine resistance in higher plants using PCR amplification of thepsbA gene coupled toMaeI digestion of the amplified product to distinguish triazine resistant from sensitive biotypes. Our assay is universal and avoids the need of lengthy procedures of previously published assays, which either required spraying of seedlings in a controlled environment, quantification of chlorophyll fluorescence of leaf discs after incubation in triazine solution, DNA sequencing of thepsbA gene, or Southern-blot analysis. Our diagnostic system is qualitative, reliable, fast and simple. More than 100 seedlings taken directly from the field can be analyzed in one day. This system has a direct application towards a more rational use of herbicides in production fields. It also represents a valuable tool to monitor spreading of resistant biotypes through time and space and can serve as a model system applicable to other gene monitoring needs.     

Parthenogenesis in an elasmobranch in the presence of conspecific males

Parthenogenesis has been observed in several elasmobranch species, primarily in public aquaria. The majority of cases of parthenogenesis have occurred either when females were held without males or once a male was removed from a female's habitat. Here we report a second instance of parthenogenesis in a zebra shark female that was housed with conspecific mature males. This study calls into question the conditions under which elasmobranch females undergo parthenogenesis.     

Prévalence de l'infection cervicale à Chlamydia trachomatis dans une population féminine consultant pour contraception.

OBJECTIVES: To determine the prevalence and risk indicators of cervical infection due to Chlamydia trachomatis among female patients consulting for contraception and to evaluate an enzyme immunoassay for the detection of C. trachomatis in this setting. DESIGN: Prevalence study. Endocervical specimens were analysed by means of culture and enzyme immunoassay. C. trachomatis infection was diagnosed through culture. SETTING: A hospital family planning clinic in Trois-Rivières, Que. SUBJECTS: All 533 female patients who consulted for contraception between November 1986 and March 1988. Results of culture were available for 495 patients. MAIN OUTCOME MEASURE: Demographic, epidemiologic and clinical information was collected by means of a standard questionnaire and a gynecologic examination. MAIN RESULTS: The prevalence rate of chlamydial infection was 9% (45/495). Enzyme immunoassay detected 37 (82%) of the infections. The mean age of the patients was 19.8 years, and 98% of the infections were diagnosed in those aged 25 years or less. The variables significantly associated with C. trachomatis infection were having more than one sexual partner in the preceding year (odds ratio [OR] 2.9; 95% confidence limits [CL] 1.7 and 5.0) and having more than one partner in the preceding 3 months (OR 2.3; 95% CL 1.2 and 4.3). These two indicators would have detected 58% and 22% of the infections respectively. CONCLUSIONS: Screening for C. trachomatis infection by means of enzyme immunoassay should be proposed to all female patients aged 25 years or less consulting for contraception in our clinic. Such screening may prove to be an effective preventive measure in other similar clinical settings.