Leukocyte-derived inhibitory activity in patients with myelodysplastic syndrome

Leukocyte-derived inhibitory activity inhibiting the entry of normal progenitor cells of granulocytes and macrophages (CFU-GM) into the S-phase of a cell cycle was investigated in 16 patients with different forms of myelodysplastic syndrome (MDS). The presence of this inhibitory activity was analysed in medium conditioned with low-density cells obtained from peripheral blood of MDS patients. The inhibition rate was measured by 3H-thymidine suicide technique with subsequent cultivation of pretreated cells in semisolid agar medium. Low-density cells from MDS patients of various types were studied: from the twelve patients with refractory anaemia (RA or RAS) only three were positive, one patient with chronic myelomonocytic leukaemia (CMML) was negative while one patient with refractory anaemia with excess of blasts (RAEB) and two patients with RAEB in transformation (RAEB-T) were positive with respect of the described test. In two patients with RA, who underwent a long-term investigation, the production of leukocyte-derived inhibitory activity preceded the development of disease into RAEB or RAEB-T. In five positive cases, supernatants were incubated with antiserum against human placental ferritin; with one exception, the inhibitory activity was neutralized.     

Cephalodiate Arten der GattungLecidea sensu lato (Ascomycetes lichenisati)

(1) The ability to produce cephalodia is usually a genus-specific character in lichens. (2)Lecidea shushanii Thoms., is a member of the genusTephromela, closely related toT. aglaea. It is not clear, whether or not the cephalodia of this taxon are true cephalodia or colonies of epiphytic cyanobacteria and whether or notLecidea shushanii is an independent species. (3)Lecidea dovrensis Nyl., is, in contrast to the traditional concept, not conspecific withLecidea alpestris Sommerf., but an earlier name forLecidea pallida Th. Fr. (4)Lecidea dovrensis is described in some detail. Chemically the species is characterized by the presence of isousnic acid (previously unknown in lecideoid lichens). It is restricted to areas north of the 60th parallel with an oceanic climate. (5) In connection with the attempt to clarify the taxonomic relationships ofLecidea dovrensis, figures of ascus apical structures of the following species are given (marked by an asterisk are genera where we found discrepancies with published data):Austrolecia antarctica, Catillaria chalybeia, Lecidea alpestris, L. caesioatra, L. limosa, Lecidoma demissum, Koerberiella wimmeriana, Micarea assimilata, M. crassipes, M. melaenida, M. prasina, Pilophorus robustus, Placodiella olivacea, Placolecis opaca, Porpidia trullisata, Protoblastenia rupestris, Psilolechia lucida, Psorula rufonigra, Squamarina gypsacea, Xanthopsorella texana. (6) Among crustaceous lichens we find no groups related toLecidea dovrensis. We supportTimal's concept of including this species in the genusPilophorus. Pilophorus, as well asLecidea dovrensis is characterized by the same ascus type, by a similar structure of thallus, cephalodia, paraphyses, and ascocarp (although there is no pseudopodetium developed inLecidea dovrensis), and the presence of isousnic acid. In addition, both taxa are restricted to cool oceanic climates and non-calciferous substrates. The following combination is proposed:Pilophorus dovrensis (Nyl.)Timdal, Hertel & Rambold, comb. nova. (7) The species of theLecidea alpestris-group form an independent genus, probably near toAustrolecia Hertel.

Frau Prof. Dr.Elisabeth Tschermak-Woess zu ihrem 70. Geburtstag gewidmet.     

Gill surface area of water-breathing freshwater fish

Summary To provide a hitherto lacking review which focuses on gill surface area of freshwater fish, we collected and analysed morphometric data from the literature. The scaling exponent of gill area ranges from 0.36 to 1.13, with a mean value of 0.76. The absolute values for the largest gill areas are about 5 times as high as those of the smallest. This range resembles that of marine fish, if specially adapted steady swimmers, such as tunnies and some sharks, are excluded. Generally it appears that the gill areas of freshwater fish are smaller than those of comparable marine species. To establish whether a relationship exists between gill area and swimming activity or oxygen content of water, the activity of each species and the oxygen content of its habitat were estimated and checked against the gill area. ANOVA revealed that activity explains the presence of the smallest gill areas only, while oxygen content does not correlate with gill area at all. The morphometric variables determining gill area (total length of filaments, average lamellar density, average lamellar area) are highly correlated; total gill area correlates mainly with lamellar density and to a lesser degree with filament length; lamellar area varies independently. Different populations of the same species exhibit striking differences with respect to gill areas, total length of filaments, average lamellar density and average lamellar area. These differences point to a substantial morphological plasticity of the gill system.     

Transferrin and iron distribution in subcellular fractions of K562 cells in the early stages of transferrin endocytosis.

Iron distribution in subcellular fractions was investigated at different times after a single cohort of 59Fe-125 I-labeled transferrin (Tf) endocytosis in K562 cells. Cell homogenates prepared by hypotonic lysis and deoxyribonuclease (DNAase) treatment were fractionated on Percoll density gradients. Iron-containing components in the postmitochondrial supernatant were further fractionated according to their molecular weight using gel chromatography and membrane filtration. In the initial phases of endocytosis, both iron and Tf were found in the light vesicular fraction. After 3 min the labels diverged, with iron appearing in the postmitochondrial supernatant and Tf in the heavy fraction containing mitochondria, lysosomes and nuclei. Iron released from Tf-containing vesicles appeared both in low- and high-molecular-weight fractions in the postmitochondrial supernatant. After 5 min of endocytosis 59Fe activity in the low-molecular-weight fraction remained constant and 59Fe accumulated in a high-molecular-weight fraction susceptible to desferrioxamine chelation. After 10 min, 59Fe radioactivity in this fraction decreased and a majority of cytosolic 59Fe was found in ferritin. These results do not support the concept of the cytosolic low-molecular-weight iron pool as a kinetic intermediate between transferrin and ferritin iron in K562 cells.     

The Larval Eye of Nannochoristid Scorpionflies (Insecta,Mecoptera)

Abstract The stemmata of last–instar Nannochoristalarvae are compound eyes composed of 10 or more ommatidia. Each ommatidium has four Semper cells, four distal and four proximal retinula cells which form a cruciform and layered rhabdom. The ommatidia are separated by epidermal cells (possibly rudimentary pigment cells). Corneal lenses are lacking. At the posterior edge, aberrant stemma units may be present which lack a dioptric apparatus and have a star–shaped rhabdom composed of at least six retinula cells. The stemmata of Nannochoristaappear to be derived from stemmata of the Panorpa-type (Mecoptera-Panorpidae). Differences between the stemmata of Nannochoristaand Panorpacan be explained as adaptations to aquatic life (flat cornea) or as regression. A compound larval eye is ascribed to the ground plan of the Mecoptera sensu latoand is considered a genuine plesiomorphy. The identical basic number (seven) of stemmata in the Neuropteroid/Coleoptera assemblage, Amphiesmenoptera and some Mecoptera (Bittacidae, Boreidae) is attributed to parallel evolution.     

Is there hybridization between diploid and tetraploid Euphrasia in a secondary contact zone?

Premise

Strong postzygotic reproductive isolating barriers are usually expected to limit the extent of natural hybridization between species with contrasting ploidy. However, genomic sequencing has revealed previously overlooked examples of natural cross-ploidy hybridization in some flowering plant genera, suggesting that the phenomenon may be more common than once thought. We investigated potential cross-ploidy hybridization in British eyebrights (Euphrasia, Orobanchaceae), a group from which 13 putative cross-ploidy hybrid combinations have been reported based on morphology.

Methods

We analyzed a contact zone between diploid Euphrasia rostkoviana and tetraploid E. arctica in Wales. We sequenced part of the internal transcribed spacer (ITS) of nuclear ribosomal DNA and used genotyping by sequencing (GBS) to look for evidence of cross-ploidy hybridization and introgression.

Results

Common variant sites in the ITS region were fixed between diploids and tetraploids, indicating a strong barrier to hybridization. Clustering analyses of 356 single-nucleotide polymorphisms (SNPs) generated using GBS clearly separated samples by ploidy and revealed strong genetic structure (F_ST = 0.44). However, the F_ST distribution across all SNPs was bimodal, indicating potential differential selection on loci between diploids and tetraploids. Demographic inference suggested potential gene flow, limited to around one or fewer migrants per generation.

Conclusions

Our results suggest that recent cross-ploidy hybridization is rare or absent in a site of secondary contact in Euphrasia. While a strong ploidy barrier prevents hybridization over ecological timescales, such hybrids may form in stable populations over evolutionary timescales, potentially allowing cross-ploidy introgression to take place.     

Progress in fold recognition

The prediction experiment reveals that fold recognition has become a powerful tool in structural biology. We applied our fold recognition technique to 13 target sequences. In two cases, replication terminating protein and prosequence of subtilisin, the predicted structures are very similar to the experimentally determined folds. For the first time, in a public blind test, the unknown structures of proteins have been predicted ahead of experiment to an accuracy approaching molecular detail. In two other cases the approximate folds have been predicted correctly. According to the assessors there were 12 recognizable folds among the target proteins. In our postprediction analysis we find that in 7 cases our fold recognition technique is successful. In several of the remaining cases the predicted folds have interesting features in common with the experimental results. We present our procedure, discuss the results, and comment on several fundamental and technical problems encountered in fold recognition. © 1995 Wiley-Liss, Inc.     

Decreased Sensitivity to Hydroxyurea and to [3H]Thymidine Suicide In the Middle of the S Phase

Pluripotent haemopoietic stem cells (CFUs) move synchronously through the cell cycle in hydroxyurea-treated mice in a cohort 1–2 hr broad. Ten to fifteen hours after hydroxyurea they pass through S phase. DNA synthesis appears to be depressed 5–10 times when the cells are in the middle part of the S phase but does not seem to be completely interrupted. High concentrations of [³H]thymidine must be used for ‘suicide’ in order to achieve lethality for the cells with depressed DNA synthesis. At the time when DNA synthesis is depressed, the sensitivity of the cells to hydroxyurea also decreases. This may lead to a significant underestimation of the S phase fraction by the hydroxyurea method, because CFUs with low DNA synthesis rate are resistant to hydroxyurea although being in S phase.