Use of tritium labeled amino acid conjugates of prostaglandins and thromboxanes as labeled ligands in prostanoid radioimmunoassay

Conjugates of prostaglandins and thromboxanes with tritium labeled amino acids were prepared and employed as labeled ligands in porstaglandin and thromboxane radioimmunoassays. Assays for PGF_2α, 15-keto-13, 14-dihydro-PGF_2α, TXB₂ and 15-keto-13, 14-dihydro-TXB₂ were evaluated in comparative studies using either these heterologous ligands or the corresponding homologus tritiated eicosanoid as tracers. Binding properties for the respective antibodies were found to be similar using either tracer.Three biological studies were also conducted, viz. study of the release of TXB₂ during collagen induced platelet aggregation, of 15-keto-13, 14-dihydro-TXB₂ during guinea pig pulmonary anaphylaxis, and of PGF_2α (measured as 15-keto-13, 14-dihydro-PGF_2α in peripheral plasma) during bovine luteolysis. The analyses gave comparable results using either the heterologous or the homologous assay.Thus, this type of labeled prostanoid conjugates may serve as a convenient alternative to homologous tracers in radioimmunoassay. Heterologous tracers may even in certain cases provide the only simple solution to the problem of preparing a labeled ligand of high specific activity.     

Inhibition of soybean lipoxygenase by SKF 525-A and metyrapone

To determine whether agents which inhibit cytochrome P-450 enzymes also inhibit lipoxygenase, the effects of metyrapone and SKF 525-A were assessed on soybean lipoxygenase using a spectrophotometric technique which allows for measurement of both the rate and magnitude of product formation. Both SKF 525-A and metyrapone inhibited the rate of product formation and the final amount of product formed in 5 min incubations SKF 525-A was 5 to 6 times more potent than metyrapone, with the IC₅₀ for SKF 525-A 40 uM and for metyrapone between 150 and 200 uM as determined by the total product formation in 5 minutes. Analysis of the reduced product by HPLC confirmed that the substances monitored were those generated by the 15-lipoxygenase enzyme.     

Microlocating lithium in the mouse embryo by use of a (n, α) nuclear reaction

Summary The quantitative imaging of lithium distribution, in histological sections of 15-days old mouse embryos (whose mother had been submitted to Li-treatment), was performed using⁶Li isotope as tracer,⁶Li(n,)³H nuclear reaction for detection, and dielectric track detectors. Despite the particular difficulties of cryosectioning the embryos without disturbing the lithium distribution, the Li regionalization appeared to be very clear-cut. The ectomesodermic tissues were significantly more loaded with lithium than the endodermic ones. This is probably related to the ectomesodermic tissues being also those most sensitive to the teratogenic effect of lithium. The Li-distribution in the embryo brain was almost homogeneous, instead of being heterogeneous as in adult brain. The mean Li-concentration in the embryo brain was not much below the Li concentration in the grey matter of the mother brain, but it was significantly larger than that in the white matter of the mother brain. Our results are discussed in the context of teratogenic effects observed in situ during mammalian development.     

DNA-polymorphic patterns linked to the β-globin genes in German families affected with hemoglobinopathies and thalassemias: a comparison to other ethnic groups

Summary DNA haplotype constellations of the β-globin gene cluster have been analyzed in German families with hemoglobinopathies (Hb Freiburg, Hb K?ln, Hb Presbyterian) and β-thalassemias. The polymorphis patterns obtained were compared to those found in families from Greece, Italy, and Turkey affected by β-thalassemia syndromes. With the combined analysis of seven restriction site polymorphisms a DNA-diagnostic prediction for additional offspring could be made with an overall frequency of 75% in the four ethnic groups.     

Effects of phosphorus and nitrogen enrichment on the phytoplankton in a tropical reservoir (Lobo Reservoir,Brazil)

The effects of enrichment with phosphate (0–500 µg. 1^–1) and forms of nitrogen (nitrate, nitrite, ammonia an and urea) (0–3500 µgg. ^–1) on the phytoplankton growth of Lobo Reservoir (Brazil) were studied in July, 1979. Suspended matter, chlorophyll a, cell concentrations and the carotenoid:cchlorophyll ratio were estimated following 14 days of in situ incubation. Phosphate alone caused no significant effects, but enrichment with nitrogen caused a substantial increase on the growth of phytoplankton. Comparison between the different forms of nitrogen showed insignificant effects after their additions with 350 µg. ^–1 and in combination with phosphate. However, when nitrogen was added in large quantities (3 500 µg. ^–1), significant differences between the nitrogeneous forms were found, with urea causing the strongest effect. In July, nitrogen is mhe main limiting nutrient to phytoplankton growth of Lobo Reservoir.Supported by CNPq and FAPESP.     

Ecological patterns of relative clutch mass in snakes

Summary Data on the relative clutch mass of snakes are summarized for over 100 populations. RCM was significantly lower in live bearing versus egg laying forms. We suggest that the longer reproductive season of viviparous snakes results in higher overall mortality compared to oviparous species; by reducing RCM, viviparous snakes may reduce this risk of mortality. Unlike lizards, no differences in RCM were found between categories of either escape behavior or foraging mode, possibly because detailed information on these behaviors are lacking for most snakes. In four populations examined, RCM did not vary among years. When compared to lizards, snakes demonstrate significantly higher RCM, perhaps owing to a more energetically efficient means of locomotion. Our data support the contention that RCM should be considered a separate and distinctive life-history characteristic of reptiles.     

Nucleotide sequence of the split tRNAUAALeu gene from Vicia faba chloroplasts: evidence for structural homologies of the chloroplast tRNALeu intron with the intron from the autosplicable Tetrahymena ribosomal RNA precursor

Summary The gene encoding the tRNA _UAA ^Leu from broad bean chloroplasts has been located on a 5.1 kbp long BamHI fragment by analysis of the DNA sequence of an XbaI subfragment. This gene is 536 bp long and is split in the anticodon region. The 451 bp long intron shows high sequence homology over about 100 bp from each end with the corresponding regions of the maize chloroplast tRNA _UAA ^Leu intron. These conserved sequences are probably involved in the splicing reaction, for they can be folded into a secondary structure which is very similar to the postulated structure of the intron from the autosplicable ribosomal RNA precursor of Tetrahymena. Very little sequence conservation is found in the 5-and 3-flanking regions of the broad bean and maize chloroplast tRNA _UAA ^Leu genes.     

Mapping of cysteine genes on the chromosome of Pseudomonas aeruginosa PAO

Summary Three loci coding for different steps in the pathway of cysteine biosynthesis have been mapped by R68.45-mediated coconjugation analysis. The cysteine auxotrophic mutants could be subdivided into sulfite and sulfide-requiring mutants. Sulfide-requiring mutants (cysIV group) were localized at a single position between pyrF and pur-67, while sulfite-requiring mutants (cysI and cysII) mapped at two different regions. The cysI group was also localized between pyrF and pur-67, although more distal to pyrF than the cysIV group. This group included the cys-54 marker, which has been mapped previously. The second group of sulfite-requiring mutants, designated as cysII, was cotransducible with hisI and localized at the end of the PAO chromosomal map. This location was also confirmed for the marker cys-59.The marker cys-59 (which was cotransducible with his1) was cotransferred by R68.45-mediated conjugations with both the late marker pur-67 and the early marker ilv-226. As the late marker hisI was positioned at about 60–65 min (Herrmann and Günther, in press) the length of the PAO chromosome was estimated to be about 70 min.     

Sequence of Guinea Pig Myelin Basic Protein

This paper proposes a tentative amino acid sequence of guinea pig myelin basic protein obtained by comparison of peptide fragments of the guinea pig and bovine proteins. Analyses of the tryptic peptides confirmed the known sequence differences in the NH2-terminal half of the molecule and showed that in the COOH-terminal half of the guinea pig protein Ser131 was missing, Ala136 - His137 was deleted, Leu140 was replaced by Phe, and an extra Ala was inserted somewhere within sequence 142-151 (tryptic peptide T23 ). Sequence determination of guinea pig tryptic peptides corresponding to residues 130-134 ( T20 ), 135-138 ( T21 ), and 142-151 ( T23 ) of the bovine protein confirmed the above sequence changes and placed the extra Ala between Gly142 and His143 . The sequence of the region corresponding to bovine residues 130-143 is thus Ala-Asp-Tyr-Lys-Ser-Lys-Gly-Phe-Lys-Gly-Ala-His. No species differences were observed in the amino acid compositions of the remaining tryptic peptides obtained from the COOH-terminal half of the molecule. Based upon these results, the guinea pig basic protein contains 167 amino acid residues and has a molecular weight of 18,256.