Die Wirkung eines Komplexbildners und von Metallionen auf die Auspr?gung dessnaky- undMonstra-Charakters beiParamaecium aurelia

Summary This paper presents new facts on the problem of the inherited defects of cell separation inParamecium aurelia (Mo-strain andsnaky, cf.Maly 1958, 1960).The complexing agent EDTA, when added to the culture medium, causes a drastic increase in monster formation in thesnaky-strain, a much smaller increase in theMo-strain and no effect in the normal strain. This effect can be compensated by the addition of different metal ions and evidence is presented that the addition of iron or cobalt to the medium normalises thesnaky- andMo-strains: the separation of the cells runs normally. It is suggested that the aldolase in thesnaky-strain and in the defective particles of theMo-strain differs from that in the normal strain.     

Molecular characterization of major histocompatibility complex class 1 (MHC-I) from squirrel monkeys (<Emphasis Type="Italic">Saimiri sciureus</Emphasis>)

Little is known about the major histocompatibility complex (MHC) class 1 in squirrel monkeys (Saimiri sciureus). We cloned, sequenced and characterized two alleles and the cDNA of the coding region of MHC class 1 in these New World monkeys. Phylogenetic analyses showed that these sequences are related to HLA class 1 genes (HLA-A and HLA-G). The structure and organization of one of the two identified clones was similar to that of a class 1 MHC gene (HLA-A2). All the exon/intron splice acceptor/donor sites are conserved and their locations correspond to the HLA-A2 gene. The sequences of the newly described cDNAs reveal that they code for the characteristic class 1 MHC proteins, with all the features thought necessary for cell surface expression. Typical sequences for the leader peptide, ₁, ₂, ₃, transmembrane and cytoplasmic domains were found.The nucleotide sequence data reported in this paper have been submitted to the GenBank database and have been assigned the accession numbers AJ438576 (Sasc-G*31), AJ438577 (Sasc-G*25), AY282760 (Sasc-G*03), AY282761 (Sasc-G*04) and AY282762 (Sasc-G*05). Sequences were named as recommended by Klein and co-workers (1990)     

The pea<Emphasis Type="Italic"> END1</Emphasis> promoter drives anther-specific gene expression in different plant species

END1 was isolated by an immunosubtractive approach intended to identify specific proteins present in the different pea (Pisum sativum L.) floral organs and the genes encoding them. Following this strategy we obtained a monoclonal antibody (mAbA1) that specifically recognized a 26-kDa protein (END1) only detected in anther tissues. Northern blot assays showed that END1 is expressed specifically in the anther. In situ hybridization and immunolocalization assays corroborated the specific expression of END1 in the epidermis, connective, endothecium and middle layer cells during the different stages of anther development. END1 is the first anther-specific gene isolated from pea. The absence of a practicable pea transformation method together with the fact that no END1 homologue gene exists in Arabidopsis prevented us from carrying out END1 functional studies. However, we designed functional studies with the END1 promoter in different dicot species, as the specific spatial and temporal expression pattern of END1 suggested, among other things, the possibility of using its promoter region for biotechnological applications. Using different constructs to drive the uidA (-glucuronidase) gene controlled by the 2.7-kb isolated promoter sequence we have proven that the END1 promoter is fully functional in the anthers of transgenic Arabidopsis thaliana (L.) Heynh., Nicotiana tabacum L. (tobacco) and Lycopersicon esculentum Mill. (tomato) plants. The presence in the –330-bp region of the promoter sequence of three putative CArG boxes also suggests that END1 could be a target gene of MADS-box proteins and that, subsequently, it would be activated by genes controlling floral organ identity.Abbreviations GUS -Glucuronidase - uidA -Glucuronidase gene - Nos Nopaline synthase gene - nptII Neomycin phosphotransferase II gene - SEM Scanning electron microscopy GenBank accession numbers for the END1 cDNA and the END1 promoter: AY 091466 and AY 324651, respectively     

Bovine β-defensins: Identification and characterization of novel bovine β-defensin genes and their expression in mammarygland tissue

-Defensin genes code for multifunctional peptides with a broad-range antimicrobial activity. In this project we hypothesized that -defensin genes may be candidate genes for resistance to mastitis. In this article we describe the identification and genomic characterization of eight bovine -defensin genes, including six novel defensin genes and two pseudogenes. Expression in the bovine mammary gland of one of the novel genes, DEFB401, has been demonstrated, as well as the expression of LAP, TAP, DEFB1, BNBD3, BNBD9, and BNBD12. For genomic characterization, 20 BACs from two different bovine BAC libraries (RZPD numbers 750 and 754) were isolated by PCR screening with -defensin consensus primers derived from published sequences. PCR products from BACs generated with consensus primers have been subcloned and sequenced, revealing a total of 16 genes and two pseudogenes. Six novel -defensin genes share the typical exon–intron structure and are highly homologous to published bovine -defensin genes. They are named DEFB401–DEFB405 and LAP-like, and two novel pseudogenes are named EBD-P and EBD-P2. Analysis of mammary gland tissue-derived cDNA from nine cows with different clinical findings demonstrated the expression of several -defensin genes mentioned above. First results indicate that the lactational status of the cow presumably has no influence on gene expression. Competent knowledge of antimicrobial activity of -defensins from literature, the abundance of -defensin mRNA in the bovine mammary gland, and the inducibility of some genes give first evidence that -defensins may play a role in local host defense during udder infections.The nucleotide sequence data reported in this article have been submitted to EMBL and have been assigned the accession numbers AJ563279–AJ563283, AJ567353–AJ567365, AJ567990–AJ567993, and AJ620296.     

Characterization of the T-cell receptor gamma locus and analysis of the variable gene segment expression in rabbit

The genomic organization and expression of genes of the T-cell receptor gamma (TRG) locus are described for mice and humans, but not for species such as rabbits (Oryctolagus cuniculus), in which T cells compose a sizeable proportion of T cells in the periphery. We cloned 200 kb of the rabbit TRG locus and determined the TRGV gene usage in adult and newborn rabbits by RT-PCR. We identified two TRGJ genes, one TRGC gene, and 22 TRGV genes, all of which encoded functional variable regions. One TRGV gene is the unique member of the TRGV2 subgroup, whereas the other genes belong to the TRGV1 subgroup. Evolutionary analyses of TRGV1 genes identified three distinct groups that can be explained by separate duplication events in the rabbit genome. Evidence of gene conversion between TRGV1.1 and TRGV1.6 was observed. Both TRGV1 and TRGV2 subgroup genes were expressed in the spleen, intestine, and appendix of adult rabbits, and the repertoire of TRGV genes expressed in these tissues was similar. In these tissues from newborns, and in skin from adults, only the genes from the TRGV1 subgroup were expressed. Greater TRGV-J junctional diversity was found in tissues from adult compared to newborn rabbits. Our analyses indicate rabbits have a larger germ line encoded TRG repertoire compared with that of mice and humans. In addition, we found TRGV gene usage is alike in most tissues of rabbits similar to that found in humans but in contrast to that found in mice.Electronic SupplementaryMaterial Supplementary material is available for this article at The nucleotide sequence data reported in this article have been submitted to GenBank and are assigned the accession numbers AY748325–AY748348     

Cytological and molecular observations on Solanum phureja-induced dihaploid potatoes

Summary Seventeen potato dihaploids, produced by pollinating the tetraploid (2n = 48) cv Pentland Crown with pollen from Solanum phureja (2n = 24) dihaploid inducer clones, were studied. Since dihaploids are thought to develop parthenogenetically from unfertilized ovules they were expected to be euploid (2n = 24), but somatic chromosome counts showed that 15 of the 17 dihaploids were aneusomatic. Ten of the clones were predominantly diploid (2n = 24) with a proportion of hyperploid cells that contained 25 or 26 chromosomes. Five of the dihaploids contained variable numbers of triploid cells (2n = 36). RFLP analysis was used to determine whether the additional chromosomes were from S. phureja or S. tuberosum. Unique hybridizing fragments present in S. phureja but not in Pentland Crown were identified. These S. phureja-specific restriction fragments were present in some of the dihaploid offspring of Pentland Crown. Of the 5 clones that contained triploid cells 4 had S. phureja type banding. Four of the 10 aneusomatic clones that contained hyperploid cells had the unique S. phureja hybridizing fragments. We propose that ovules of Pentland Crown were fertilized by pollen from S. phureja and that the aneusomatic clones were derived from triploid zygotes from which some of the S. phureja chromosomes were eliminated. We consider that this is an additional mechanism of dihaploid formation in potato.     

Characterization of the horse (<Emphasis Type="Italic">Equus caballus</Emphasis>)<Emphasis Type="Italic"> IGHA</Emphasis> gene

Nucleotide sequences of the immunoglobulin constant heavy chain genes of the horse have been described for IGHM, IGHG and IGHE genes, but not for IGHA. Here, we provide the nucleotide sequence of the genomic IGHA gene of the horse (Equus caballus), including its secretion region and the transmembrane exon. The equine IGHA gene shows the typical structure of a mammalian IGHA gene, with only three exons, separated by two introns of similar size. The hinge exon is located at the 5 end of the CH2 exon and encodes a hinge region of 11 amino acids, which contains five proline residues. The coding nucleotide sequence of the secreted form of the equine IGHA gene shares around 72% identity with the human IGHA1 and IGHA2 genes, as well as the bovine, ovine, porcine and canine IGHA genes, without distinct preference for any of these species. The same species also cluster together in a phylogenetic tree of the IGHA coding regions of various mammals, whereas rodent, rabbit, marsupial and monotreme IGHA genes each build a separate cluster.The nucleotide sequences reported in this paper have been assigned the EMBL/GenBank accession numbers AY247966 and AY351982     

The partial amino acid sequence of a murine Ia molecule

Eleven of the amino terminal 14 amino acid residues have been assigned in the chain of the murine I-C^d subregion molecule. The murine chain shows no homology with P29 (the putative human chain equivalent), the chain of guinea pig Ia. 4, 5, or the murineI-A subregion chain.     

Assessment of the Respiratory Costs of Adaptation in Plant Species That Differ in Their Responses to Insufficient and Excessive Mineral Nutrition

The effects of insufficient and excessive mineral nutrition on the relative growth rate (RGR); the rate of total dark respiration (R); gross photosynthesis (Pg); and the contents of endogenous IAA, ABA, and cytokinins (CK) were studied in 20–60-day-old seedlings of plant species that differed in their responses to these factors: beet root (Amaranthus retroflexusL.), orchard-grass (Dactylis glomerataL.), and common wheat (Triticum aestivumL.). The changes in the level of mineral nutrition lowered the RGR and Pgvalues and the IAA/ABA and CK/ABA ratios and increased the Rand R/Pgvalues and ABA content, especially in A. retroflexusand T. aestivum. The authors propose a new way to assess the respiratory cost of adaptation to stress based on the relation between the RGR and the R/Pgvalues. The adaptation component of Ris shown to relate to the ABA content.     

Effect of mutant host RNA polymerase on the bifunctional activities of P22 gene c1.

Summary A phage has been isolated which specifically transduces the Escherichia coli pheS and pheT genes coding for the and subunits of the phenylalanyl-tRNA synthetase (PRS). This phage transduces with high frequency (i) several temperaturesensitive PRS mutants to thermoresistance and (ii) a p-fluorophenylalanine resistant PRS mutant to sensitivity against this amino-acid analog. The in vitro PRS activities of such lysogens suggest that the and subunits coded by the transducing phage complement the mutant host PRS-subunits in vivo by means of formation of hybrid enzymes.The transducing phages were also used to infect UV light irradiated cells. The SDS-gel electrophoretic analysis of the proteins synthesized in such cells revealed that the phage codes at least for four different E. coli proteins. Two proteins with molecular weights of 94,000 and 38,000 daltons cross-reacted with an anti PRS serum and were thus identified as the and subunits of PRS, respectively. A third protein with w molecular weight of 22,000 daltons is identical with the ribosomal initiation factor IF3 (Springer et al., 1977b). The other protein (M_r 78,000) is still unidentified.     

Phenylalanyl-tRNA synthetase fromE. coli MRE-600: Localization of the phenylalanine binding sites on the subunits by affinity reagents

The phenylalanine and the phenylalanyl-tRNA^Phe binding sites on the subunits of phenylalanyl-tRNA synthetase fromE.coli MRE-600 were localized using p-azidoanilidate of [¹⁴C]phenylalanine and N-bromoacetyl[¹⁴C]phenylalanyl-tRNA^Phe. The phenylalanine recognizing site was shown to be situated on the subunit of the enzyme in close proximity to the contact region of the and subunits and the phenylalanyl-tRNA^Phe recognizing site on the subunit. Transfer of the aminoacyl moiety from the subunit to the subunit of the enzyme was assumed to take place in the process of catalysis of the aminoacylation reaction.     

Discrimination of sunflower volatiles by the red sunflower seed weevil

Five principle monoterpenoid and other constituent volatile chemicals of sunflower heads were combined to resemble two lines of sunflower (Helianthus annuus L.): one U.S.D.A. standard line and one French line which was poorly visited by insects (Etievant et al., 1984). Field trials of attraction to red sunflower seed weevils (Smicronyx fulvus Le Conte, Coleoptera: Curculionidae) showed that one was clearly preferred over the other. The more attractive mixture contained -pinene, -pinene, limonene, camphene and bornyl acetate in a ratio resembling that of Flath et al. (1985) rather than that described by Etievant et al. (1984). One or two volatiles were deleted from the optimal blend but only mixtures of five volatiles showed the highest attraction. Substitution of sabinene, another volatile prominent in sunflower, for one of the five in the optimal blend also decreased attraction of seed weevils. When the monoterpenoid components and green leaf volatiles in the traps resembled the ratios of most of the prominent volatiles of sunflower, attraction was significantly greater than controls.     

Feeding Behavior, Habitat Use, and Abundance of the Angelfish Holacanthus passer (Pomacanthidae) in the Southern Sea of Cortés

Feeding behavior and habitat use of the king angelfish, Holacanthus passer, was studied in the southern Sea of Cortés, México. H. passer fed on benthic communities (algae and sessile invertebrates) and in the water column (mainly feces from the damselfish Chromis atrilobata). Although there were not significant differences in feeding rate between sexes, coprophagy was more common in males, while grazing was more common in females. Spatial distribution of size classes followed a pattern of decreasing size with increasing depth. Feeding rate was significantly different among habitats: small females had a higher feeding rate on the bottom, big females and small males had similar feeding rates from the bottom to 3m above the bottom, and big males had higher feeding rates from 5m above the bottom to the surface. Habitat was clearly partitioned, and there was significant habitat overlap only between big females and small males. The abundance of H. passer was partly explained (34% of the total variance) by the abundance of the damselfish C. atrilobata. There was a clear trophic association between C. atrilobata schools and H. passer feeding damselfish feces in the water column. The sex ratio male:female of H. passer populations was >1 at several sites, an unusual pattern for a protogynous fish. The sex ratio on the H. passer water column stock was also biased towards males at most sites. Although there is a positive relationship between C. atrilobata abundance and H. passer, there are factors other than damselfish abundance which cause this dominance of males.     

Segmental duplication associated with the human-specific inversion of chromosome 18: a further example of the impact of segmental duplications on karyotype and genome evolution in primates

The human-specific pericentric inversion of chromosome 18 was analysed using breakpoint-spanning BACs from the chimpanzee and human genome. Sequence and FISH analyses disclosed that the breakpoints map to an inverted segmental duplication of 19-kb, which most likely mediated the inversion by intrachromosomal homologous recombination. The 19-kb duplication encompasses the 3 end of the ROCK1 gene and occurred in the human lineage. Only one copy of this segment is found in the chimpanzee. Due to the inversion, the genomic context of the ROCK1 and USP14 genes is altered. ROCK1 flanks USP14 in the long arm of the chimpanzee chromosome 17, which is homologous to human chromosome 18. This order is interrupted by the inversion in humans. ROCK1 is localized close to the pericentromeric region in 18q11 and USP14 is inverted to distal 18p11.3 in direct neighbourhood to LSAU-satellites, -satellites and telomere-associated repeats. Our findings essentially confirm the analysis of Dennehey et al. (2004). Intriguingly, USP14 is differentially expressed in human and chimpanzee cortex as well as fibroblast cell lines determined previously by the analysis of oligonucleotide arrays. Either position effects mediated by the proximity to the telomeric region or nucleotide divergence in regulatory regions might account for the differential expression of USP14. The assignment of the breakpoint region to a segmental duplication underlines the significance of the genomic architecture in the context of genome and karyotype evolution in hominoids.     

Biofilm thickness effect on the diffusion limitation in the bioprocess reaction: Biofloc critical diameter significance

A convenient physical model for biomass involves consideration of individual cells as active centers dispersed through a continuous region in which transport takes place by molecular diffusion. This paper investigates for steady state conditions the variation of apparent kinetic constant (K _m ) of bacteria in relation to biofloc diameter with solid and/or liquid-phase diffusion. When the biochemical reactions are limited only by liquid-phase diffusion, theK _m increases whenD increases. With solid-phase diffusion limitation only, theK _m increases linearly with the diameterD of the floc. When both solid and liquid-phase diffusion limitations are considered, the apparentK _m is affected by liquid-phase diffusion limitation with very smallD and by solid-phase diffusion limitation with higherD. The critical diameterD _c can be assumed to be theD at which solid-phase diffusion limitation becomes more significant than liquid-phase diffusion limitation.     

Characterization of sodium and pyruvate interactions of the two carrier systems specific of mono- and di- or tricarboxylic acids by renal brush-border membrane vesicles

Summary The experiments reported in this paper aim at characterizing the carboxylic acid transport, the interactions of pyruvate and citrate with their transport sites and specificity. The study of these carriers was performed using isotopic solutes for the influx measurements in brush-border membrane vesicles under zerotrans conditions where the membrane potential was abolished with KCl preloading with valinomycin or equilibrium exchange conditions and =0.Under zerotrans condition and =0, the influence of pyruvate concentrations on its initial rates of transport revealed the existence of two families of pyruvate transport sites, one with a high affinity for pyruvate (K _t =88 m) and a low affinity for sodium (K _t =57.7mm) (site I), the second one with a low affinity for pyruvate (K _t =6.1mm) and a high affinity for sodium (K _t =23.9mm) (site II). The coupling factor [Na]/[pyruvate] stoichiometry were determined at 0.25mm and 8mm pyruvate and estimated at 1.8 for site I, and 3 when the first and the second sites transport simultaneously.Under chemical equilibrium (0) single isotopic labeling, transport kinetics of pyruvate carrier systems have shown a double interaction of pyruvate with the transporter; the sodium/pyruvate stoichiometry also expressed according to a Hill plot representation wasn=1.7. The direct method of measuring Na⁺/pyruvate stoichiometry from double labeling kinetics and isotopic exchange, for a time course, gives an=1.67.Studies of transport specificity, indicate that the absence of inhibition of lactate transport by citrate and the existence of competitive inhibition of lactate and citrate transports by pyruvate leads to the conclusion that the low pyruvate affinity site can be attributed to the citrate carrier (tricarboxylate) and the high pyruvate affinity site to the lactate carrier (monocarboxylate).     

Concluding remarks

On the basis of symposium contributions onChlorella, Hibbertia, Eucalyptus, Ambrosia and on numerical approaches some fundamental problems of (bio)systematics, evolution, and taxonomic categories are discussed: Methods available for analysing affinities; conflicting evidence from phenetic, biochemical, cytogenetic and other analyses; further classification problems in cases of intermediacy, etc. While sibs of various levels and their natural hierarchy often can be objectively defined, this appears impossible for particular taxonomic levels itself (e. g. species). A single objective taxonomic system of organisms is unrealistic. Certain guiding lines for relative and practicable concepts of species and genus are proposed.Presented at the symposium Speciation and the Species Concept during the XIIth International Botanical Congress, Leningrad, July 8, 1975.     

Aggression in damselfish: habituation by adults of Pomacentous parritus to juvenile intruders

Synopsis Successful recruitment of juvenile coral reef fishes may depend, in part, upon the aggressive behavior of adults already on the reef. In addition to initial levels of aggression, changes in aggressive behavior of adults, e.g., due to habituation, may have an even greater influence on recruiting juveniles. Adult males of the bicolor damselfish, Pomacentrus partitus, were used as subjects to study habituation of aggression toward conspecific and congeneric (P. variabilis) juveniles. Adults, held in a 1000 liter aquarium, habituated after 4 hours of constant exposure to juveniles restrained in a 1 liter model bottle. Stimulus strength of the juveniles depended on their species identity, size and proximity to the resident adult's shelter. There was a recovery of aggression with a change in stimulus location, but at the same location, a stimulus of greater strength was required to bring about recovery. The implications of these findings for coral reef fish community structure are discussed.     

Effects of mutations in genesfadR,fabB, fadE,andenvC ofEscherichia coli on the action of the lysis gene of bacteriophageϕX174

The lytic effect of the expression of the cloned geneE of bacteriophage X174 inEscherichia coli is considerably amplified by a mutation in thefadR gene, which primarily affects the regulation of fatty acid degradation. In contrast, reduction of the fluidity of the cell membranes by use of thefabB andfadE mutations, which interfere with the synthesis and the oxidation of unsaturated fatty acids, severely inhibits the action of the X174 lysis gene product. A chain-forming mutant carrying a pleiotropic mutation in theenvC locus is also refractory to the X174 lysis protein. As shown by reversion and complementation of theenvC mutatation, a defect in at least one additional gene (rle) is involved in the generation of this refractoriness.     

Thermodynamic and kinetic considerations of Q-cycle mechanisms and the oxidant-induced reduction of cytochromesb

In coenzyme Q-cycles, it is proposed that one electron from the quinol reduces the Rieske iron sulfur center (E _m280 mV) and the remaining electron on the semiquinone reduces cytochromeb _T (E _m–60 mV). TheE _mfor the two-electron oxidation of the quinol is 60 mV and therefore the reduction of cytochromeb _T by quinol is not favorable. As the stability constant for the dismutation of the semiquinone decreases, the calculatedE _mfor the Q/QH couple is lowered to values below theE _mof cytochromeb _T. Contemporary coenzyme Q-cycles are based on the belief that the lower value for theE _mof the Q/QH couple compared to theE _mfor cytochromeb _T means that the semiquinone is a spontaneous reducing agent for theb-cytochrome. The analysis in the paper shows that this is not necessarily so and that neither binding sites nor ionization of the semiquinoneper se alters this situation. For a Q-cycle mechanism to function,ad hoc provisions must be made to drive the otherwise unfavorable reduction of cytochromeb _T by the semiquinone or for the simultaneous transfer of both electrons to cytochromeb _T and cytochromec ₁ (or the iron sulfur protein). Q-cycle mechanisms with these additional provisions can explain the observation thus far accumulated. A linear path which is functionally altered by conformational changes may also explain the data.