Involvement of nuclear factor-kappaB in endothelin-A-receptor-induced proliferation and inhibition of apoptosis

Endothelins have been implicated in the regulation of cell proliferation, differentiation, and apoptosis, but the mechanisms of these complex events are not yet fully understood. Although the nuclear factor-B (NF-B) was shown to play a prominent role in the above processes, its participation in endothelin receptor A (ET_AR) signaling has not been previously demonstrated. This study provides evidence that NF-B is involved in ET_AR-induced proliferation and inhibition of apoptosis. Endothelin (ET)-1, ET-3, and sarafotoxin b induce cell proliferation and prevent apoptosis induced by serum deprivation in a Chinese hamster lung (CCL39) cell line that stably expresses ET_AR (CCL39ET_A). Activation of ET_AR resulted in enhanced DNA-binding activity of NF-B and degradation of IB-. Expression of the dominant negative form of IB- (IBN) inhibited the proliferative activities mediated by ET_AR as well as its antiapoptotic activities. Treatment of the cells with prostaglandin A₁, an inhibitor of IB kinase-, reduced ET-1-induced proliferation and its antiapoptotic effect. These findings indicate that the regulation of cell proliferation and apoptosis by ET_AR is mediated by the ET_AR-activated NF-B.     

The enthalpies of interactions of some L-α-amino acids with urea molecule in aqueous solutions at 298.15 K

Summary. Dissolution enthalpies of L--aminobutyric acid, L--isoleucine, L--phenylalanine, L--methionine, L--serine, L--threonine, L--cysteine, L--asparagine and L--glutamine in aqueous solutions of urea have been measured by calorimetry at 298.15K. The obtained results were used to calculate the enthalpic interaction coefficients between the zwitterions of the L--amino acids and a molecule of urea in water. These values were interpreted in terms of the hydrophobic or hydrophilic effects of the side chains of amino acids on their interactions with a polar molecule of urea in water.     

Mapping the α-globin genes in HB J Mexico carriers

Summary the organization of the -globin genes was studied by restriction endonuclease mapping, in subjects carrying the variant Hb J Mexico. A subject homozygous for Hb J synthesized both Hb J (about 55%) and Hb A and had two loci per chromosome. His restriction site map was found to be identical to that obtained with a normal DNA, except for a mutant Bgl II site which was observed on the Hb J chromosome proximal to the 5-locus. We have also mapped the DNA of a compound heterozygote for Hb J and -thalassemia, who synthesizes 38% Hb J and we have found a single gene corresponding to a –^3.7 haplotype on one chromosome and two genes, respectively ^J and ^A, on the other.     

Emission of gaseous nitrogen oxides from an extensively managed grassland in NE Bavaria, Germany.

We analysed the stable isotope composition of emitted N₂O in a one-year field experiment (June 1998 to April 1999) in unfertilized controls, and after adding nitrogen by applying slurry or mineral N (calcium ammonium nitrate). Emitted N₂O was analysed every 2–4 weeks, with additional daily sampling for 10 days after each fertilizer application. In supplementary soil incubations, the isotopic composition of N₂O was measured under defined conditions, favouring either denitrification or nitrification. Soil incubated for 48 h under conditions favouring nitrification emitted very little N₂O (0.024 mol g_dw ^–1) and still produced N₂O from denitrification. Under denitrifying incubation conditions, much more N₂O was formed (0.91 mol g_dw ^–1 after 48 h). The isotope ratios of N₂O emitted from denitrification stabilized at ¹⁵N = –40.8 ± 5.7 and ¹⁸O = 2.7 ± 6.3. In the field experiment, the N₂O isotope data showed no clear seasonal trends or treatment effects. Annual means weighted by time and emission rate were ¹⁵N = –8.6 and ¹⁸O = 34.7 after slurry application, ¹⁵N = –4.6 and ¹⁸O = 24.0 after mineral fertilizer application and ¹⁵N = –6.4 and ¹⁸O = 35.6 in the control plots, respectively. So, in all treatments the emitted N₂O was ¹⁵N-depleted compared to ambient air N₂O (¹⁵N = 11.4 ± 11.6, ¹⁸O = 36.9 ± 10.7). Isotope analyses of the emitted N₂O under field conditions per se allowed no unequivocal identification of the main N₂O producing process. However, additional data on soil conditions and from laboratory experiments point to denitrification as the predominant N₂O source. We concluded (1) that the isotope ratios of N₂O emitted from the field soil were not only influenced by the source processes, but also by microbial reduction of N₂O to N₂ and (2) that N₂O emission rates had to exceed 3.4 mol N₂O m^–2 h^–1 to obtain reliable N₂O isotope data.     

Tunneling Processes Induced by Terahertz Electric Fields

Tunneling processes induced by terahertz frequency electric fields havebeen investigated.A drastic enhancement of the tunneling probabilityhas been observed by increasing the frequency at_e 1 where_e is the tunneling time.For a given constant tunneling rate an increase offrequency by a factor of seven leads to a drop of the requiredelectric field strengthby three orders of magnitude.It is shown that the enhancement of tunneling ionization at terahertz frequencies is due to the factthat electrons can absorb energy from the radiation field during tunnelingreducing the effective width of the tunneling barrier.     

C to T and/or G to A transitions are responsible for loss of a MspI restriction site at the 5′-end of the human apolipoprotein AI gene

We detected the loss of a MspI restriction site by a C to T transition at +83 bp and a G to A transition at +84 bp of the 5-end non-coding region of the human apolipoprotein AI gene. This base change occurred at the hot spot (CCGG) for methylation, which may be important in the regulation of gene expression. The population frequency for the loss of the MspI site is 6.1%.     

Heterogeneity of the α-globin gene defects in German α-thalassemia affected families

Summary Analysis of -thalassemia syndromes in several German families revealed DNA deletion as well as nondeletion forms as the molecular basis for the defects. Thus, the -thalassemia haplotype was identified as the (–)^3.7 rightward deletion form, and the region of the putative recombination process generating such a deletion was further characterized. In addition three different ° haplotypes, (--)^MED, (--)^>26, and ()^T, could be detected using -and -globin gene-specific probes.     

Restriction fragment length polymorphism of the human T cell receptor alpha gene

Previous studies have demonstrated restriction fragment length polymorphisms (RFLP) in the vicinity of the alpha and beta genes of the human T-cell receptor. In the course of experiments designed to discover additional polymorphic restriction sites, we found a new RFLP of the T-cell alpha gene recognized by the restriction enzyme Taq I. The site was localized to the interval between the most 3 joining (J) exon and the most 5 constant (C) region exon, about 7 kb distant from the previously described Bgl II polymorphic site which mapped to the vicinity of the 3 untranslated exon. With the use of these two polymorphic markers, four Ti-alpha alleles could be identified, allowing unambiguous assignment of all Ti-alpha genes in some families. These markers may be useful in identifying possible immune response genes or disease predisposition genes associated with the genes of the T-cell receptor for antigen.Abbreviations used in this paper RFLP restriction fragment length polymorphism - Ti-alpha alpha gene of the T-cell receptor for antigen     

The sea anemone genus <Emphasis Type="Italic">Actinostola</Emphasis> (Verrill 1883): variability and utility of traditional taxonomic features,and a re-description of <Emphasis Type="Italic">Actinostola chilensis</Emphasis> (McMurrich 1904)

Species of the genus Actinostola are known for high variability of features. Anatomy, histology and cnidae of type specimens of five species from South America and Antarctica originally described as members of Actinostola and one species of Stomphia were compared to specimens of Actinostola chilensis collected during this study. None of these traditionally used features clearly distinguish the examined Actinostola species. I therefore propose new distinctive taxonomic features, including in vivo and in situ data. I provide an emended diagnosis of the genus Actinostola and a revised list of its species. I accept the synonymy of A. excelsa, A. pergamentacea and A. intermedia with A. crassicornis, and reject the synonymy of A. chilensis with A. crassicornis and A. intermedia. I re-describe A. chilensis in detail, including in situ information. Specimens of A. chilensis inhabit exposed positions of rocky substrate from 22 m depth down in south Chilean fjords between Puerto Montt (41°3535S, 72°53W) and Puyuhuapi (44°3136S; 72°326W); the most conspicuous features are its relatively large size, bright-orange colour, smooth, tough column and numerous and clearly entacmaeic tentacles.Electronic Supplementary MaterialSupplementary material (appendices) is available for this article at An erratum to this article can be found at     

Electrolytes control flows of water across the apical barrier in toad skin: The hydrosmotic salt effect

Summary The reversible dependence of skin osmotic water permeability (L _PD ) upon the ionic concentration of the outer bathing solution — which we have called hydrosmotic salt effect (HSE) — was studied in the isolated skin of the toadBufo marinus ictericus. The skin osmotic water flow (J _V ) was measured as a function of outer bathing solution osmolality (O _e ).L _PD , calculated as (J _v /) _P=0 (where and P are the osmotic and hydrostatic pressure differences across the skin, respectively) was constant whenO _e was altered with sucrose, a nonelectrolyte. In contrast,L _PD increased continuously in the hypotonic range asO _e was raised from zero (distilled water) with NaCl or KCl. The HSE could also be evoked in the condition of reversed osmotic volume flow, with the outer bathing medium made hypertonic with sucrose.Diffusional¹⁴C-sucrose permeability, measured in theJ _v =0 condition to prevent solvent drag of sucrose in the paracellular pathways, indicate that the hydrosmotic salt effect cannot be explained by assuming a paracellular permeability increase, due to tight junction opening, but might be interpreted as due to changes in the osmotic water permeability of the apical membranes of the most superficial cells of the epithelium.The hydrosmotic salt effect can be elicited in control skins and in vasopressin-stimulated skins, on top of the hormonal response.The time course of the hydrosmotic salt effect is substantially different from that of the hydrosmotic response to vasopressin. Its half-time is 4 to 5 times faster than that of vasopressin action, with individual values as short as 1.5 min.The time courses of the hydrosmotic salt-effect onset and reversibility are exponential, clearly contrasting with the typical sigmoidal shape of vasopressin onset and washout time courses.Based on time course data and on speed of response we postulate that the mechanism underlying the hydrosmotic salt effect is due to modifications of existing water pathways in the apical membrane, rather than to incorporation and removal of water permeability units in this structure.     

Genetic Linkage Maps of the Guppy (Poecilia reticulata): Assignment of RAPD Markers to Multipoint Linkage Groups

Genetic linkage maps of the guppy (Poecilia reticulata) were constructed from independent crosses between the Tuxedo strain and a feral line (Wildtype). Segregation patterns of random amplified polymorphic DNA (RAPD) markers and phenotypic markers were investigated in F₂ offspring of Tuxedo × Wildtype and Wildtype × Tuxedo crosses. Among the 300 and 276 RAPD markers scored for the respective crosses, linkages were identified for 230 and 212, respectively. The Tuxedo × Wildtype and Wildtype × Tuxedo maps spanned 2100 Kosambi centiMorgans (cM_K) and 1900 cM_K, respectively, in 28 linkage groups. Average marker resolution was 10 cM_K. Genome length was estimated at 4410 cM_K and 4060 cM_K for the respective crosses, with an average physical distance of 166 kbp/cM_K. Several RAPD markers were closely linked to or mapped onto the loci for the sex-determining region (SdR), and the sex-linked black caudal-peduncle (Bcp) and red tail (Rdt) genes. These primary linkage maps are the initial step toward the construction of a composite high-density map to facilitate map-based cloning and marker-assisted selection of quantitative trait loci that are essential for the development of comprehensive breeding programs for the guppy.     

Functional characterisation of two cytochrome<Emphasis Type="Italic"> b</Emphasis><Subscript>5</Subscript>-fusion desaturases from<Emphasis Type="Italic"> Anemone leveillei</Emphasis>: the unexpected identification of a fatty acid Δ<Superscript>6</Superscript>-desaturase

The Ranunculaceae are known to accumulate a wide range of unusual fatty acids in their seed lipids, and this variability has been advocated as a taxonomic marker. The Anemone species, Anemone leveillei L. and Anemone rivularis Buch.-Ham., have previously been reported to accumulate ⁵-desaturated fatty acids in their seed tissue [K. Aitzetmüller (1995) Plant Syst Evol 9:229–240]. Two cDNAs, AL1 and AL2, with similarity to plant cytochrome b₅-fusion "front-end" desaturases were isolated from developing seeds of A. leveillei and their function identified by expression in Saccharomyces cerevisiae. AL2 was characterised as a sphingolipid long-chain-base ⁸-desaturase, while AL1 acted as a fatty acid desaturase. However, AL1 did not produce ⁵-desaturated fatty acids as expected; instead, when expressed in transgenic S. cerevisiae or Arabidopsis thaliana this enzyme was functionally characterised as a ⁶-desaturase. Northern analysis confirmed the expression of this gene in seed tissue and leaf tissue of A. leveillei, though ⁶-desaturated fatty acids were found to accumulate only in the leaf tissue. The unexpected characterisation of a ⁶-desaturase in A. leveillei has implications for the use of fatty acids in chemotaxonomic studies. This is also the first report of a higher-plant ⁶-desaturase from a family other than the Boraginaceae.Abbreviations ALA -linolenic acid - DMOX 4,4-dimethyloxazoline - EDA eicosadienoic acid - FAME fatty acid methyl ester - GLA -linolenic acid - LA linoleic acid - LCB long chain base - ORF open reading frame - OTA octadecatetraenoic acid     

The good,the bad and the ugly: science,aesthetics and environmental assessment

The question is raised, whether there are peculiarly scientific values which can be applied in environmental assessment. The use of the expression scientific interest is traced from its 19th century origins to modern British statutes. It is argued that attempts to replace expert judgements by objective scientific criteria (e.g., indices of biodiversity) can never be completely successful. In particular, interest is an aesthetic atribute particularly valued by scientists but incapable of precise measurement. While science provides the best framework for informed judgements on conservation issues, the judgements of scientists are inevitably distinct from their experimental results. Judgements rest on ethical and aesthetic values such as importance and interest, which are essential constituents of the scientific sub-culture, but which are not uniquely scientific.     

In vitro effects of monoterpenes on chloroplast membranes

A chloroplast preparation was extracted from squash (Cucurbita pepo (L.) var. Senator). Enrichment of intact chloroplasts was achieved by continuous free-flow electrophoresis. The addition of monoterpenes, detergent and free fatty acids changed the elecrophoretic separation pattern characteristically. Monoterpene-dependent degradation of envelope membranes could be prevented by addition of -tocopherol prior to monoterpene incubation.Photosynthetic electron transport of photosystem II was completely inhibited by -pinene, Triton X-100 and linolenic acid. Inhibition could be modulated by addition of -tocopherol or lecithin (phosphatidylcholine) either before or after inhibition by monoterpenes and detergent.Percentage reconstitution of photosynthetic electron transport inhibited by -pinene depended on light conditions and incubation time.     

‘Small’-proteoglycan: collagen interactions: Keratan sulphate proteoglycan associates with rabbit corneal collagen fibrils at the ‘a’ and ‘c’ bands

l. Proteoglycans (PGs) in rabbit corneal stroma and mouse sclera have been stained for electron microscopy with Cupromeronic blue in a critical electrolyte concentration (CEC) mode, with and without prior digestion of the tissue by keratanase or chondroitinase ABC to remove the keratan sulphate (KS) or chondroitin-dermatan sulphates (CS or DS) respectively.2. Two classes of PGs, located orthogonally to the corneal collagen fibrils at either the step (band a or c) or gap zone (band d or e) are shown to be KS-PGs or DS-PGs respectively. Four separate and specific PG binding sites on Type I collagen fibrils have thus been identified.3. Rabbit corneal KS and DS PGs each contain two kinds of PG (Gregory JD, Coster L & Damle SP (1982) J. Biol. Chem.257, 6965–6970). We propose that each small protein-rich PG is associated with a specific binding site on the collagen fibril.     

Carotenoid composition in milo (Sorghum vulgare) shoots as affected by phytochrome and chlorophyll

The composition of coloured carotenoids in the milo shoot was investigated quantitatively (high performance liquid chromatography) during light-mediated plastidogenesis, including the time span of photodelay as caused by medium and high light fluxes. It was found that as long as only the far-red-absorbing form of phytochrome operates, the carotenoid pattern remains virtually the same as in complete darkness (violaxanthin and lutein as major constituents, traces of -carotene). On the other hand, the pattern changes dramatically in white or red light with increasing amounts of chlorophyll (lutein and -carotene dominate, -carotene showing the strongest relative increase). Photodelay during the early phase of plastidogenesis affects the carotenoid composition strongly. Increase of neoxanthin, violaxanthin and -carotene contents are diminished while lutein accumulation proves resistant towards chlorophyll-mediated photoinhibition. The photodelay can be diminished by an appropriate light pretreatment. The data indicate that light-mediated control over carotenoid accumulation is exerted at three levels: i) a coarse control through phytochrome, ii) fine tuning in connection with chlorophyll accumulation, iii) stabilization of holocomplexes against photodecomposition.Abbreviations GG14 high fluence rate green-yellow light - HPLC high-performance liquid chromatography - Chl chlorophyll - WL_w weak white light (1200 lx) - WL_m medium flux white light (12000 lx)     

The butterfly and the serpent: Culture,psychopathology and biomedicine

Cultural explanations of psychopathology in the West have rarely employed models derived by anthropologists for small-scale non-literate communities. Some general features of those ritual patterns usually classed as culture-bound syndromes are applicable to Western neurosis. Such reactions articulate both personal predicament and public concerns. usually core structural oppositions between age groups or the sexes. They gain their power. by relying on certain unquestionable assumptions which. although beyond everyday jural relationships, articulate such relationships. In the case of Western reactions, such mystical sanction is provided by biomedicine. Theoretical paradigms emphasize either the individual pragmatic or expressive aspects, or social homeostasis.     

Evidence for G-protein-linked receptors in higher plants: stimulation of GTP-gamma-S binding to membrane fractions by the mastoparan analogue mas 7

Stimulation of G-protein activity by the amphipathic tetradecapeptide mastoparan is well documented in animal systems and occurs via mimickry of the third cytoplasmic domain of the cognate seven-transmembrane-span (7TMS) receptor. Binding of guanosine 5-O-thiotriphosphate to microsomal and plasma-membrane fractions from Pisum sativum L. and Zea mays L. was stimulated by the tetradecapeptide mastoparan analogue mas 7 over a narrow concentration range, with maximal effect exerted at 10 M peptide, while the nonamphipathic analogue, mas CP, which differed from mas 7 by only one amino acid, was ineffective at promoting binding. This stimulation could be completely abolished by inclusion of low concentrations of the non-ionic detergents nonanoyl-N-methylglucamide and Lubrol PX. Taken together, these data provide good evidence for the existence of G-protein-linked 7TMS receptors in higher plant systems.Abbreviations GTPS guanosine 5-O-thiotriphosphate - MEGA-9 nonanoyl-N-methylglucamide - 7TMS 7 trans-membrane span The authors wish to acknowledge the financial support of the European Community (I.R.W.), ICI and the Science and Engineering Research Council (A.W.) and the Agricultural and Food Research Council (P.A.M.).     

Chemical and physiological changes in fungi during autolysis

A comparative study was done on some of the chemical changes occurring during autolysis of cultures ofAspergillus flavus in both physiologically acid and alkaline media. The mycelium ofA. flavus lost during autolysis 44 % of its maximum dry weight in the physiologically alkaline medium, whereas this loss was apparently nil in the physiologically acid medium. Nitrogen containing compounds seemed not to be affected by autolysis either in the physiologically acid or alkaline media. The disappearance of P-containing compounds in mycelium ofA. flavus autolysed in both conditions (NO ₃ ^– and NH ₄ ⁺ as N source) amounted to 64 % in the alkaline autolysis and to nearly 77 % in the acid autolysis. The results we have obtained for the acid autolysis strongly suggest that very little activity is shown by autolytic enzymes in the interval 10–133 days of incubation, when measuring autolysis by the loss in mycelial dry weight.

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Zusammenfassung Eine vergleichende Untersuchung war unternommen an einigen der chemischen Veränderungen, die während der Autolyse der Kulturen vonAspergillus flavus in physiologischen sauren und alkalischen Medien vorkommen. Die Myzelien vonA. flavus haben während der Autolyse 44 % ihres größten Trockengewichtes in physiologisch alkalischem Medium verloren, während dieser Verlust in physiologisch saurem Medium anscheinend Null gewesen ist. Stickstoff enthaltende Substanzen erschienen während der Autolyse weder in physiologisch saueren noch in alkalischen Medien beeinflußt zu sein. Das Verschwinden von P-enthaltenden Substanzen in Myzelien vonA. flavus in Autolyse unter beiden Bedingungen (NO ₃ ^– und NH ₄ ⁺ als Stickstoffquelle) erreichte 64 % in alkalischer Autolyse und beinahe 77 % in der saueren Autolyse. Die Ergebnisse, die wir in der saueren Autolyse erhalten haben legen es sehr nahe, daß autolytische Enzyme eine sehr geringe Aktivität in der Zeitspanne von 10–133 Tagen der Inkubazion zeigen, wenn die Autolyse an dem Verlust des mycelialen Trockengewichtes gemessen wird.

     

Importance of intermediate disturbances for the species composition and diversity of phytoplankton in two very different Berlin lakes

Species composition and diversity of phytoplankton were studied for several years in two lakes which differ with respect to mixing conditions and nutrient limitation: Schlachtensee regularly stratifies very stably. In contrast, size and wind-exposure predispose Lake Tegel to deeper mixing; additionally, stratification is artificially destabilized by aeration. As the duration of aeration was varied, the study period includes interannual changes in mixing conditions. For both lakes, it also covers trophic change due to restoration; this was especially pronounced in Schlachtensee.Results show that mixing conditions affect species composition on two levels: on a superordinate level, lake morphology or hydrology govern stability of stratification and susceptibility to perturbation, and hence the extent to which motile species can develop. In Schlachtensee, species with some means of actively seeking preferred depths usually dominated during summer stratification: Planktothrix agardhii during the hypertrophic phase, and flagellates since restoration. In contrast, in Lake Tegel deeper mixing as a generally prevailing condition favored non-motile species. Their seasonal pattern was remarkably constant from year to year. Although changes in the extent of mixing were pronounced during the four years studied (1987–1990), these were within a range that affected species composition only slightly: in summer, cyanobacteria and diatoms represented climax species whose dominance was not offset by additional, weather-induced increases of turbulence.On a subordinate level, and within the constraints set by nutrient limitation as well as by grazing pressure, small-scale changes in mixing conditions caused by meteorological cycles were shown to strongly affect species composition and in consequence diversity: Results for the fouryear post-restoration study period at Schlachtensee show that considerable interannual variations of species composition and diversity can be attributed to variations in the frequency of meteorological changes. In accordance with the intermediate disturbance hypothesis (IDH), diversity was lowest during 1989, the year with the longest and most pronounced cycles of fair weather (14 to 27 fair days on end). However, the mechanism for this was rarely a decline of diversity caused by competitive exclusion within single long phases of stable conditions, as conceived by the intermediate disturbance hypothesis. Instead, diversity responded to changes in mixing conditions with a variety of patterns — often with low values during phases of increased mixing and with high values under quiescent conditions, especially during the first calm days just after increased mixing. Thus, not disturbance as such, but rather the rate of change between phases of disturbance and quiescence appears to determine the frequency of high diversity indices.In Lake Tegel, high diversity indices were somewhat more frequent in 1989, the year during which thermal stratification was most stable. For species adapted to frequent or continuous mixing, interjected calm phases with unusually high stability of thermal stratification may represent a disturbance of accustomed conditions. Thus, in turbulent Lake Tegel, meteorological cycles appear to act in reversal to the IDH, in a sense which may be termed intermediate quiescence hypothesis.Phosphorus limitation due to successful restoration was found to decrease winter and vernal diversity in Schlachtensee. Presumably, without nutrient constraints, new populations could grow more rapidly in response to the rapid changes of physical parameters during this season. In contrast, restoration has increased summer diversity, as phosphorus concentrations no longer allow the virtual monocultures of Planktothrix agardhii which prevailed previously. In Lake Tegel, the reduction of phosphorus concentration down to 60 µg/l P in 1989 limited biomass, but this level was still too high to significantly alter species composition or diversity.