Untersuchungen zur Inkompatibilitt imCulex pipiens-Komplex

Zusammenfassung Bei Kreuzungen zwischenCulex pipiens-Popalationen verschiedener geographischer Herkunft werden drei Kreuzungstypen festgestellt: normale Kreuzbarkeit, reduzierte Kreuzbarkeit und Inkompatibilitt (Nichtkreuzbarkeit). Die drei Kreuzungstypen sind mit Hilfe der Embryonierungsrate, der Schlüpfrate und der entstehenden Nachkommenschaft gegeneinander abgrenzbar. Bei Inkompatibilitt sind 99,9% der Embryonen letal, und etwa 0,1% der Tiere schlüpfen und sind fertile, diploide Weibchen.Die Aktivierung des Eies und der Entwicklungsansto\ erfolgt durch das Spermium. Das Spermium gelangt nicht zur Karyogamie mit dem Pronucleus. Es liegt induzierte, meiotische Parthenogenese vor.Die diploiden, parthenogenetischen Weibchen gehen aus einer Oocyte 2. Ordnung oder aus Teilungsprodukten einer Oocyte 2. Ordnung hervor. Die letalen Embryonen sind haploid. Das Spermium beteiligt sich nicht am Aufbau des Embryos. Nach Aktivierung des Eies wird das Spermium im Eiplasma resorbiert, whrend der haploide Pronucleus einen rein haploiden Embryo aufbaut.Bei einigen Kreuzungen entwickeln sich bis zu 75% der Embryonen bis zum Stadium der histologischen Differenzierung der Organe. Die Embryonen zeigen noch einige Stunden nach dem normalen Schlüpftermin Muskelkontraktionen, vermögen die Eihülle jedoch nicht zu sprengen.Die DNS-Verteilungen von Interphasekernen der letalen Embryonen liegen zwischen den Werten C und 2C für haploide Zellen. ltere Embryonen besitzen in geringem Ma\e höhere DNS-Werte als 2C. Whrend der histologischen Differenzierung der Organe liegen in diploiden Kontrollembryonen und in letalen Embryonen die Ploidiestufen C, 2C, 4C, 8C and 16C vor. Bei inkompatiblen Kreuzungen wird das in das Ei eindringende Spermium cytophotometrisch nachgewiesen. Die Wanderung des Spermiums im Ei wird untersucht.In Normalkreuzungen wird nur ein geringer Grad von Polyspermie festgestellt. Monound Dispermie sind am hufigsten. Es werden die Möglichkeiten diskutiert die zur Blockierung der Karyogamie in inkompatiblen Kreuzungen führen können.

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Investigations on the incompatibility in theCulex pipiens-complex

Summary In crosses between populations of the mosquitoCulex pipiens of different geographical origin three crossing types have been found (1) crosses with normal offspring (2) crosses with reduced offspring and (3) crosses that show almost total incompatibility. In the case of incompatible crosses 99.9% of the embryos are lethal and only about 0.1% of the embryos hatch and develop to fertile diploid females. Based on genetical and cytological data it is argued that induced meiotic parthenogenesis takes place. The sperm does not play any part in the production of the diploid females and the lethal embryos. After the activation of the egg the sperm moves to the center of the egg but it does not succeed in fusing with the pronucleus. As a result the pronucleus starts to develop into a haploid embryo in about 99.9% and only in a few cases the diploidy is restored by a change in the meiotic process in the egg. Up to 75% of the haploid embryos develop to the stage of histological differentiation. The frequency distribution of the DNA in interphase nuclei of these embryos shows a maximum at C and 2 C characteristic for haploid cells. The absence of ploidy classes higher than 2 C in the early embryos is in agreement with the assumption of pure haploidy. After histological differentiation ploidy classes C to 16 C can be found in tissues that show endomitotic growth. The development of the haploid embryos is described. It has been shown through cytophotometric methods that in incompatible crosses entrance of the sperm into the egg takes place. In normal crosses polyspermy is rather rare, monospermy and dispermy are most common. The blocking of the sperm in incompatible crosses is discussed.

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Teil einer Dissertation der Math.-Nat. Fakultt der Universitt Mainz. Herrn Prof. Dr. H. Laven danke ich für die Bereitstellung des Untersuchungsmaterials sowie für die Anregungen bei der Durchführung der Arbeit.     

FEMORAL SHAFT FRACTURES—A Study of Closed Reduction and Open Treatment

A comparative study was made of 58 cases of closed femoral shaft fractures treated by skeletal traction, and 24 cases of closed femoral shaft fractures treated by open reduction with internal fixation.Although complications occurred in some cases, intramedullary nailing appeared to be the most satisfactory method, resulting in primary union, in decreased time of recumbency and time in hospital, in earlier ambulation and in less residual disability.Success of intramedullary nailing depends largely upon adequate training or experience of the surgeon in the technical operative aspects of the procedure and in postoperative management.Placing supplemental autogenous iliac bone chips at the fracture site in closed femoral fractures in which intramedullary nailing is performed appears to enhance callus formation and bony consolidation.Skeletal traction should be utilized on all patients whose general physical condition does not permit operative intervention.     

THE CELLULAR COMPLEMENT OF THE SKELETAL SYSTEM STUDIED AUTORADIOGRAPHICALLY WITH TRITIATED THYMIDINE (H3TDR) DURING GROWTH AND AGING

An autoradiographic study has been made of mouse femora from birth to one year of age. Tritiated thymidine was used to study the proliferative rate of various skeletal tissues. The labeling index of the periosteum was found to be highest at birth (0.085). At 8 weeks of age the index fell sharply to 0.007 and remained low throughout the entire period of study. Chondro-osteogenic cells of the periosteum, especially those at the perichondrial zone, were most frequently labeled initially. The labeling index of the epiphyseal disk remained high (0.054 to 0.048) until about the 6th week of age. Following this age the index fell sharply to 0.018 at 8 weeks and 0.002 at 26 weeks of age. Autoradiographs showed that the cells of the articulating surface of the epiphysis and disk are derived at least in part from migrating chondro-osteogenic cells of the periosteum residing at the perichondrial region, and that these cells serve as the progenitor pool necessary for both circumferential growth and expansion of the epiphyseal disk.     

An estradiol-dependent protein from chicken liver binds single-stranded DNA and RNA

We have identified two estradiol-dependent single-stranded DNA binding proteins in the nucleus and cytoplasm of chicken hepatocytes that bind the sequence 5'TCACCTTCGCTATG3' in the first exon of the chicken vitellogenin gene. As judged by chromatography on heparin-Sepharose and by proteolytic clipping bandshift assay, the two proteins are different. Furthermore, they only bind to the oligonucleotide corresponding to the upper strand. Depurination and depyrimidination interference experiments with the cytoplasmic protein show that the bases CCTT-G are involved in the protein-DNA interaction. An RNA corresponding to the upper strand of the gene between nucleotide positions -73 and +53 competes for binding to the single-stranded DNA. UV cross-linking experiments performed with bromouridine-substituted single-stranded RNA reveal that an estradiol-dependent hepatocyte cytoplasmic protein with a Mr of 71,000 binds to the mRNA-like single-stranded RNA.     

Enzymes Involved in Anaerobic Polyethylene Glycol Degradation by Pelobacter venetianus and Bacteroides Strain PG1

In extracts of polyethylene glycol (PEG)-grown cells of the strictly anaerobically fermenting bacterium Pelobacter venetianus, two different enzyme activities were detected, a diol dehydratase and a PEG-degrading enzyme which was characterized as a PEG acetaldehyde lyase. Both enzymes were oxygen sensitive and depended on a reductant, such as titanium citrate or sulfhydryl compounds, for optimal activity. The diol dehydratase was inhibited by various corrinoids (adenosylcobalamin, cyanocobalamin, hydroxocobalamin, and methylcobalamin) by up to 37% at a concentration of 100 μM. Changes in ionic strength and the K⁺ ion concentration had only limited effects on this enzyme activity; glycerol inhibited the enzyme by 95%. The PEG-degrading enzyme activity was stimulated by the same corrinoids by up to 80%, exhibited optimal activity in 0.75 M potassium phosphate buffer or in the presence of 4 M KCI, and was only slightly affected by glycerol. Both enzymes were located in the cytoplasmic space. Also, another PEG-degrading bacterium, Bacteroides strain PG1, contained a PEG acetaldehyde lyase activity analogous to the corresponding enzyme of P. venetianus but no diol dehydratase. Our results confirm that corrinoid-influenced PEG degradation analogous to a diol dehydratase reaction is a common strategy among several different strictly anaerobic PEG-degrading bacteria.     

Characterization of the eaeA gene from rabbit enteropathogenic Escherichia coli strain RDEC-1 and comparison to other eaeA genes from bacteria that cause attaching-effacing lesions

Abstract A number of enteric pathogens, including enteropathogenic (EPEC) and enterohemorrhagic (EHEC) Escherichia coli , Hafnia alvei , a strain of Citrobacter freundii , and rabbit EPEC strain RDEC-1 cause attaching-effacing (AE) lesions in the gut mucosa. These bacteria have a pathogenicity cassette (locus of enterocyte effacement or LEE) containing the eaeA gene. This gene encodes intimin, an outer membrane protein required for production of AE lesions. RDEC-1, a non-invasive enteropathogen in young rabbits, produces AE lesions morphologically indistinguishable from lesions caused by human AE bacterial strains. The RDEC-1 example of E. coli diarrhea in rabbits is an important model for studying the pathogenesis of AE bacteria in a natural infection and for analyzing specific roles of the components of LEE. In order to better understand the role of intimin in the development of AE lesions, a portion of DNA within RDEC-1 LEE, containing the eaeA gene and an upstream open reading frame (ORF), was sequenced. The RDEC-1 eaeA gene shared 87%, 92%, and 93% DNA sequence identity and > 80% amino acid sequence identity with the eaeA genes of C. freundii biotype 4280, EHEC O157:H7, and EPEC O127:H6, respectively. The carboxy-terminal 280 amino acid residues of intimin has 80%, 56%, and 54% identity with C. freundii , EHEC O157:H7, and EPEC O127:H6 intimins, respectively. The predicted protein encoded by the upstream ORF (156 amino acids) shares 95%, 97%, and 99% amino acid identity with predicted proteins from C. freundii , EHEC O157:H7, and EPEC O127:H6, respectively. The high degree of sequence homology of the ORF and the eaeA gene of RDEC-1 with those of other AE bacteria suggests an evolutionary relationship of LEE and supports and facilitates the use of the RDEC-1 model for studying the role of LEE in pathogenesis.     

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.