Linkage analysis of the Duffy blood group marker with several chromosome 1 genes in an extended pedigree with Charcot-Marie-Tooth disease

Summary We have recently demonstrated tight linkage of the Duffy blood group marker to the -spectrin gene in an extended pedigree with Charcot-Marie-Tooth neuropathy. To determine a more precise location of the Duffy blood group locus on the chromosome 1 map we have tested several more chromosome 1 genes for linkage with this marker. We found suggestive linkage with the antithrombin III and apolipoprotein A2 genes and conclusive linkage with the gene coding for -nerve growth factor.     

In Vivo and In Vitro Evidence for the Biosynthesis of Testosterone in the Telencephalon of the Female Frog

Abstract: Neurons and glial cells are capable of synthesizing various steroid hormones, but biosynthesis of testosterone in the CNS has never been reported. The aim of the present study was to demonstrate the synthesis of testosterone in the frog brain. The presence of 17β-hydroxysteroid dehydrogenase (17β-HSD)-like immunoreactivity was detected in a population of glial cells located in the telencephalon. Reversed-phase HPLC analysis of brain tissue extracts combined with radioimmunoassay detection revealed the presence of substantial amounts of testosterone and 5α-dihydrotestosterone (5α-DHT) in the telencephalon where 17β-HSD-positive cells were visualized. In male frogs, castration totally suppressed testosterone and 5α-DHT in the blood and in the rhombencephalon but did not affect the concentration of these two steroids in the telencephalon. Chemical characterization of testosterone in female frog telencephalon extracts was performed by coupling HPLC analysis with gas chromatography-mass spectrometry. Using the pulse-chase technique with [³H]pregnenolone as a precursor, the formation of a series of metabolites was observed, including dehydroepiandrosterone, androstenedione, testosterone, 5α-DHT, and estradiol. These data demonstrate the existence of an active form of 17β-HSD in the frog telencephalon, which is likely involved in testosterone biosynthesis within the brain.     

A proenkephalin A-derived peptide analogous to bovine adrenal peptide E from frog brain: Purification, synthesis, and behavioral effects

A peptide derived from the posttranslational processing of proenkephalin A was isolated from an extract of the brain of the European green frog Rana ridibunda and its primary structure established as: Tyr-Gly-Gly-Phe-Met-Arg-Arg-Val-Gly-Arg¹⁰-Pro-Glu-Trp-Trp-Gln-Asp-Tyr-Gln-Lys-Arg²⁰-Tyr-Gly-Gly-Phe-Met. The structure was confirmed by chemical synthesis. The peptide represents an amphibian equivalent of bovine adrenal peptide E [preproenkephalin A (206–230)-peptide] but the sequence contains two amino acid substitutions (Met¹⁵ → Gln and Leu²⁵ → Met) compared with the mammalian peptide. The data support previous hypotheses that the Leu-enkephalin sequence is not present in preproenkephalin A of amphibians. Intracerebroventricular injections of frog peptide E (10 and 100 ng) in mice had no significant effect on horizontal locomotor activity. The peptide, in doses up to 1 μg, had no effect on latency of escape jumping in the hot plate test and the peptide (100 ng) did not modify responses (paw licking, rearing, and escape jumping) in morphine-treated mice.     

Changes in distribution of chloroplast adenylate kinase isoforms during floral induction

In the short day plant Chenopodium rubrum and the long day plant Nicotiana tabacum cv. Havana 425, adenylate kinase (EC 2.7.4.3) occurs as a family of isoforms, with at least two members localized in the chloroplast representing the main isoforms. In this work, isoforms were separated by anion exchange chromatography and relative isoform activities were compared between vegetative plants and plants induced to flowering. In both species examined, a light regime leading to floral induction resulted in a significant decrease in the activity of one chloroplast isoform. This decrease modified considerably the relative distribution of isoform activities, especially that between the two chloroplast activities.     

A protein binds the selenocysteine insertion element in the 3'-UTR of mammalian selenoprotein mRNAs.

Several gene products are involved in co-translational insertion of selenocysteine by the tRNA(Sec). In addition, a stem-loop structure in the mRNAs coding for selenoproteins is essential to mediate the selection of the proper selenocysteine UGA codon. Interestingly, in eukaryotic selenoprotein mRNAs, this stem-loop structure, the selenocysteine insertion sequence (SECIS) element, resides in the 3'-untranslated region, far downstream of the UGA codon. In view of unravelling the underlying complex mechanism, we have attempted to detect RNA-binding proteins with specificity for the SECIS element. Using mobility shift assays, we could show that a protein, present in different types of mammalian cell extracts, possesses the capacity of binding the SECIS element of the selenoprotein glutathione peroxidase (GPx) mRNA. We have termed this protein SBP, for Secis Binding Protein. Competition experiments attested that the binding is highly specific and UV cross-linking indicated that the protein has an apparent molecular weight in the range of 60-65 kDa. Finally, some data suggest that the SECIS elements in the mRNAs of GPx and another selenoprotein, type I iodothyronine 5' deiodinase, recognize the same SBP protein. This constitutes the first report of the existence of a 3' UTR binding protein possibly involved in the eukaryotic selenocysteine insertion mechanism.     

The effect of nutrient supplementation on the biofiltration removal of butanal in contaminated air

Butanal is one of the odorous compounds produced in the animal-rendering and food-processing industries and also in sewage-treatment plants. It shows the necessity for complementing such plants with systems for off-gas treatment. Biofiltration using simple packing material was tested for the removal of butanal. Excellent results were obtained when the filters operated at optimal humidity and were supplemented with inorganic nutrients. Without nutrients, butyric acid was detected in the effluent gas, which may explain the lower efficiency of filters without nutrients. Under optimal conditions an elimination of around 90 g·m^–3·h^–1 was reached.     

Substance P-Evoked Calcium Mobilization and Ionic Current Activation in the Human Astrocytoma Cell Line U 373 MG: Pharmacological Characterization

Abstract— In the human astrocytoma cell line U 373 MG, application of substance P (SP) leads to a transient increase in cytosolic calcium concentration and to a biphasic current response in voltage-clamped cells. Using these two functional assays we have characterized pharmacologically the SP response in U 373 MG cells. SP and [l -Pro⁹]SP displayed high potencies in both assays with EC₅₀values of 2.5 ± 10^?9M and 1 ± 10^?9M on calcium responses and 110^?9M and 510^?9M on ion current responses, respectively. The high potency of SP and [l -Pro⁹]SP as well as the low potency of [Lys⁵,MeLeu⁹,N-Leu¹⁰]neurokinin A_(4-10) and the inactivity of senktide demonstrate the NK₁-type pharmacology of these responses. Furthermore, the NK₁ antagonists (±)-CP 96,345, its chloro analogue, (±)-cis-3-(2-chlorobenzylamino)-2-benz-hydrylquinuclidine, and RP 67580 were potent antagonists of both SP responses. For the calcium mobilization induced by SP (1 (10^?7M), the IC₅₀ values for the three antagonists were 4 ± 10^?10M, 4 ± 10^?9M, and 9 ± 10^?9M, respectively, whereas on the current response evoked by SP 10^?8M), the IC₅₀ values were 8 ± 10^?9M, 2.4 ± 10^?8M, and 1.2 10^?7M, respectively. Despite differences in the absolute IC₅₀ values obtained with both techniques, the relative potencies of the three antagonists correlate fairly well. The U 373 MG cell line provides a useful model system for studies of the pharmacology of the human NK₁receptor and its transduction mechanisms at the level of second messengers and modulation of ion currents.     

The 23-kilodalton E1 phosphoprotein of bovine papillomavirus type 1 is nonessential for stable plasmid replication in murine C127 cells.

The 23-kDa protein encoded by the 5' segment of the E1 open reading frame of bovine papillomavirus type 1 (BPV1) was previously ascribed a negative regulatory function for the replication of viral plasmid DNA. However, results from recent functional and biochemical studies do not readily support this genetic assignment. Therefore, we have reassessed the role of this protein in papillomavirus DNA replication by using a mutant of BPV1 which is unable to express this E1 protein. This mutant viral DNA was found to replicate extrachromosomally with stability and copy number per cell similar to those of wild-type plasmid DNA. Thus, the absence of expression of the 23-kDa E1 protein did not lead to deregulated viral plasmid replication. We conclude that the 23-kDa E1 protein is nonessential for stable plasmid replication.     

Quantitative analysis of abscisic acid in needles of Abies alba Mill. by electron capture gas chromatography

Summary The amount of abscisic acid (ABA) in needles of silver fir from a natural location was investigated with regard to position in the crown, damage, seasonal variation, and needle age. Because of problems of quantification of ABA in coniferous needles, which contain numerous secondary plant products, a method for reliable determination of both isomers cis-trans-ABA (c-ABA) and transtrans-ABA (t-ABA) was developed. By means of gas chromatography (GC) using an electron capture detector (BCD) and a programmed temperature vaporizer (PTV) injector complete separation of both compounds was achieved. Two different pairs of fir were investigated — in each case a damaged and a healthy tree. Needles from both trees from the first and the second pair collected in September contained 500–1100 ng c-ABA/g fresh weight (FW), and the concentrations of t-ABA varied from 400 to 700 ng/g FW. Investigations from the second pair show highest amounts of 2900 ng/g Fw c-ABA and 1800 ng/g FW of t-ABA in May and June. For the first pair a higher c-ABA content was found in needles from the top of the crown than in those from the middle and the base. This difference could not be confirmed in the analysis of the second pair. Because of the strong natural deviation no statistically significant difference between the healthy and the damaged tree was found. The first pair of firs examined showed a higher t-ABA concentration than the second one. In this case the highest amount was found in the top of the crown. Methodical mistakes during the clean-up procedure and in quantification by gas chromatography could be excluded. The presence of c- and t-ABA in the purified extract was corroborated by mass spectrometry. With regard to the seasonal variation both isomers of ABA show an unequivocal trend. The maximum concentration is achieved in May to June, whereas the content is minimal in August/September. In any case the level of t-ABA is lower than that of c-ABA. No correlation between the amount of ABA and the needle age could be established.     

Beobachtungen am Nest des Zwergschn?ppers(Muscicapa parva)

Zusammenfassung Im Buchenwald des Naturschutzgebietes Serrahn (Mecklenburg) achtete Verf. seit 1949 auf den Zwergschnäpper. 1956 und 1957 brüteten dort über 12 Paare.Ankunft frühestens am 9. Mai. Als Erste erscheinen vorwiegend ältere (rotkehlige) . Den ersten folgen sehr bald die ersten .Balzflug, Zeigen der Nisthöhle, Copula sowie Variationen des Neststandes werden beschrieben.Fast stets baute nur das , in einem Falle 3 Tage lang. Während der Bauzeit setzt das seine Singflüge eifrig fort. Gepaarte verstummen, sobald ihr begonnen hat, fest auf dem Gelege zu brüten. Nur Junggesellen singen den ganzen Sommer über.An einem günstig gelegenen Nest wurde das Verhalten des Paares vom 4.(?) Bruttag ab bis zum Ausfliegen der Jungen (im Alter von 13 Tagen) aus einem dicht davor angebrachten Schirm genau beobachtet und aufgezeichnet. Nach dem Verlassen des Nestes suchen die Jungen unter Führung ihrer Eltern die nächste Dickung auf. Sobald sie selbständig geworden sind, beginnt die Jugendmauser, 3 Wochen danach die Wanderung ins Winterquartier. Die letzten Zwergschnäpper verschwanden meist vor Mitte September.Angaben über Gesang, Bedeutung der Rufe, Nahrung.Attrappen-Versuche am Nest ergaben, daß die Eltern genau zwischen dem (gefährlichen) Sperber und dem (ungefährlichen) Kuckuck zu unterscheiden wußten und ihnen gegenüber unterschiedliche Alarmlaute brachten. Tannenhäher und Wacholderdrossel lösten keine Abwehr-Reaktion aus.