Molecular ecology of a Neolithic meadow: The DNA of the grass remains from the archaeological site of the Tyrolean Iceman

The paper reports on the molecular analysis of samples of approximately 5,300-year-old grass found at the alpine archaeological site where the so-called Tyrolean Iceman was discovered. The grass comes from a cloak made of long grass blades and/or the stuffing of the snow footwear worn by the Iceman. The results show that while the largest fraction of the DNA extractable from the grass is of foreign origin, a much smaller part belongs to the original genetic material of the grass itself, and can be used as a valuable taxonomic clue to the plant species utilized by neolithic men to manufacture their equipment. On the other hand, the foreign DNA, or at least a portion of it, comes from microorganisms-mainly filamentous fungi and unicellular algae-which seem to have been associated with the grass since the time the grass was harvested.     

Melatonin decreases the amplitude of the b-wave of the human electroretinogram

In a double-blind placebo crossover study of 13 healthy volunteers, the pineal hormone melatonin (10 mg) was given at 4 pm, and the electroretinogram measured under conditions of dark and light adaptation. A significant diminution of b-wave amplitude was found under both photopic (=5.4 V, p<0.05) and scotopic conditions (=7.4 V, p<0.01). These data indicate that melatonin may transduce the dark signal at the level of the retina as well as the pineal. Acute administration of melatonin decreases sensitivity to light.     

Epidermal growth factor (EGF) accelerates the maturation of fetal mouse intestinal mucosa in utero

Summary Pregnant Swiss ICR mice were injected i.p. with 0.5 g of epidermal growth factor (EGF) per g b. wt at 15, 16 and 17 days of gestation and fetuses were removed at 18 days of gestation. EGF treatment had no effect on the weight of the fetuses and on the length of the small intestine. No modification of the protein and DNA contents was noted. However brush border alkaline phosphatase and trehalase activities were significantly increased as well as endoplasmic reticulum membrane-bound glucose-6-phosphatase.Supported by grant MA-6069 from the Medical Research Council of Canada. Dr D. Ménard is a chercheur boursier du Fonds de la recherche en santé du Québec     

Über die «absoluten» Bewegungen der Milchstrasse und der nächsten Spiralnebel

Summary (1) The milky way is moving absolutely (relative to the light-ether) with a velocity of at least 600 km/s in the direction of the absolute movement of the earth (A=78°,D=+40°).(2) The point of symmetry of the absolute velocities of the spirals is situated about 1,120,000 parsec from the milky way in the direction of the antapex of his absolute movement, and it has taken 1·83 milliards of years for the milky way to cross this distance.(3) Probably the next spirals are forming with the milky way as a center a group with a common parallel movement like that one of the milky way.     

Determinant of efficiency of a monomeric enzyme: Acceleration by site-specific molecules for trypsin

Summary The interaction of a specific ligand at substrate binding site was shown to be responsible for the catalytic efficiency of trypsin. The reasoning of induced fit theory was refined by kinetic analysis of characteristic properties of inverse substrates.This work was supported by grants from the Ministry of Education, Science and Culture, Japan.     

Evidence for the extranuclear localization of thymosins in thymus

A new radioimmunoassay has been developed for thymosin ₄ by generating rabbit polyclonal antibodies against the synthetic N-terminal peptide fragment 1–15 coupled to KLH. The synthetic analogue [Tyr¹²]-thymosin ₄ (1–15) was used as tracer. This radioimmunoassay, with a useful range of 10–1000 pmoles, showed cross-reactivity with the second homologous -thymosin of man and rat (thymosin ₁₀) but not of calf (thymosin ₉). This radioimmunoassay, together with an improved radioimmunoassay for the N-terminus of parathymosin , was employed for the measurement of the levels of thymosin ₄ and parathymosin in nuclear and extranuclear extracts of calf thymus. The bulk of these polypeptides was found in the extranuclear material whereas only traces were observed in the nuclear environment, which indicates the extranuclear localisation of - and -thymosins.     

Studies onxHaynaldoticum sardoum Meletti et Onnis metabolism during seed life-span: α-amylase and glutamate decarboxylase activity

Summary Embryo GAD activity and -amylase in the endosperm of 2 different physiological lines (CP; CV) ofxHaynaldoticum sardoum Meletti et Onnis were evaluated and different stages of seed ripening and progressively older seeds were examined. Results concerning GAD activity during ripening show differences between CP and CV seeds, the former being more active. In the ageing seeds, the GAD remains constant (CP is twice as much as CV) up to 4th year and greatly decreases at the 5th. The -amylase activity is fairly constant during ripening in CV endosperm and increases in CP: at the fully-ripe stage, both show similar values. During seed ageing, the activity decreases progressively in CV endosperm while, in CP, values are greater but fairly constant. The results are discussed in connection with dormancy and the different physiological ageing of seeds.This work was supported by a grant from the Consiglio Nazionale delle Ricerche (Rome, Italy).Acknowledgments. The authors wish to thank Mr F. Saviozzi, Mr V. Sbrana and Mr R. Bertini for their expert technical assistance.     

Defensive steroids from a carrion beetle (Silpha americana)

Summary The defensive anal effluent discharged bySilpha americana in response to disturbance contains a mixture of steroids stemming from a glandular annex of the rectum. The compounds have been characterized as 15-hydroxyprogesterone (1, principal component), 5-pregnan-15-ol-3, 20-dione (2), 5-pregnan-3, 15-diol-20-one (3), 5-pregnan-7, 15-diol-3,20-dione (4), 5-pregnan-3, 7, 15-triol-20-one (5), 5-pregnan-16-ol-3,20-dione (6), and 5-pregnan-3, 16-diol-20-one (7), none previously found in insects. Bioassays with jumping spiders showed compounds1 and6 to be feeding deterrents at the 1 g level.Paper No. 78 of the series: Defense Mechanisms of Arthropods. Study supported by NIH (Grants AI-02908, AI-12020). We thank Maura Malarcher and Maria Eisner for excellent technical help, and the Squibb Institute of Medical Research for samples of authentic reference compounds.     

Exchange of tritium from randomly tritiated taurocholate by microbial bile salt oxidoreductases

Summary Tritium distribution on randomly labelled taurocholate (TC) was estimated at 28%, 4%, 1% and <0.5% on the hydrogens opposite the 3-, 7- and 12-OH groups and taurine moiety respectively. Anomalously,C. perfringens 3-hydroxysteroid dehydrogenase (3-HSDH) catalyzed tritium loss of 36% on formation of 7-, 12-dihydroxy-3-keto-5-cholanoate, implying additional losses of tritium at other sites by this enzyme.This work is supported by the National Cancer Institute.     

Potential juvenile hormone analogues containing piperidine ring: Synthesis and biological assay on insects

Summary Some compounds containing piperidine rings have been synthesized, and submitted to biological tests on insects for their hormonal activity on pupae ofTenebrio molitor L. andBombyx mori L. Some of them display a hormoneanalogue or toxic activity.The editors do not hold themselves responsible for the opinions expressed in the authors' brief reports. — Les auteurs sont seuls responsables des opinions exprimées dans ces brèves communications. — Für die Kurzmitteilungen ist ausschliesslich der Autor verantwortlich. — Per le brevi comunicazioni è responsabile solo l'autore. — . — Solo los autores son responsables de las opiniones expresadas en estas comunicationes breves.This research has been supported by the Italian National Research Council (CNR) special ad hoc program Fitofarmaci e Fitoregolatori, Subproject. 7.Acknowledgments. We thank Prof. G. Jommi for helpful discussions related to this and previous articles on the same subject.     

Studies of a key protein in the mechanism of the excitation-contraction coupling process of frog skeletal muscle,using phenylglyoxal

The excitation-contraction (E-C) coupling process in single twitch fibres from frog toe muscle was inhibited selectively by phenylglyoxal (PGO), a specific guanidyl modifying reagent. A new protein (31.5 kDa), which has PGO-binding ability and seems to play a key role in the E-C coupling process, was solubilized from transverse tubule membrane-junctional sarcoplasmic reticulum complexes (TTM-JSR) of frog skeletal muscles, using¹⁴C-PGO. The monoclonal antibody against this protein applied extracellularly inhibited the E-C coupling process of the single fibres. This protein appears to constitute the very first step of input for E-C coupling. It is considered to behave as an indispensable part of an electrometer to measure membrane potentials. Therefore, the name electrometrin is suggested for the new protein.     

High-affinity binding ofβ-alanine to cerebral synaptosomes might involve glycine-receptors

Summary High-affinity, Na⁺-independent binding of -alanine to a synaptosomal fraction of rat brain was potently inhibited by glycine and by some other -amino acids, but not by taurine or GABA. This binding mechanism, which was also sensitive to both bicuculline and strychnine, might involve synaptic receptors for both -alanine and glycine.This study was supported in part by Centro Ramón y Cajal and Fundacion Juan March.     

Casein,a prohormone with an immunomodulating role for the newborn?

Summary Maternal colostrum and milk, the earliest food of the newborn, should not only be considered as supplying nutrients, but also as agents providing protection against aggressions from the new environment. Indeed by enzymatic digestion of the main milk proteins, the caseins, biologically active peptides are released; they may be implicated in the stimulation of the newborn's immune system. From this point of view a strategic active zone has been characterized in -casein. A possible role of casein as a prohormone for the newborn is suggested.     

Phase-shift and memorization of the circadian rhythm of transpiration ofTamarix aphylla

Summary The control of the circadian rhythm of transpiration inTamarix aphylla seems to include two distinct components: an externally induced one, that is initiated by the light on signal, and an endogenous clock, whose memorization of the period length is independent of the instant environmental signal.This investigation was carried out at the Tel Aviv University and supported by the MINERVA foundation and by the DFG.     

The role of repair in radiobiology

Summary Apart from cancer and mutation induction, radiobiological effects on mammals are mostly attributable to cell death, defined as loss of proliferative capacity. Survival curves relate retention of that capacity to radiation dose, and often manifest a quasi-threshold (shoulder). The shoulder is attributable to an initial mechanism of repair (Q-repair) which is gradually depleted as dose increases. Another form of repair, which is not depleted (P-repair), increases the dose required to deliver an average of one lethal event per cell (dose D₀). Neither form of repair can unambiguously be linked with repair of defects in isolated DNA. An important initial lesion may well be disruption of the complex structural relationship between the DNA, nuclear membrane and associated proteins. One form of P-repair may be restoration of that structural relationship.     

Interdependence between locomotor activity and duration of wakefulness in humans during isolation

Summary Locomotor activity was recorded in 14 subjects who lived singly in an isolation unit for 16 to 88 days. Their free-running circadian rhythms had a mean period of 25.9 h, with individual means in the duration of wakefulness () ranging from 12.1 to 22.9 h. Intraindividually, the hourly means of activity were negatively correlated with to such a degree that the total amount of activity per day remained constant irrespective of large variations in .     

Host macrophages are involved in systemic adoptive immunity against tumors

Summary The positive systemic therapeutic results obtained with adoptive transfer of immune spleen cells could not be reproduced in macrophage depleted mice. Thus, host macrophages are involved in systemic adoptive immunity against tumors.This study was supported by the Rijksuniversiteit Utrecht and the Netherlands Cancer Society (Konigin Wilhelmina Fonds, KWF).     

Différence d'activité oestrogène entre l'α-bromotriphényléthylène et un de ses analogues pentadeutérié

Summary The oestrogenic activity of -bromo-,-diphenyl--pentadeuteriophenylethylene is compared with that of the non-deuterated analogue, and found to be significantly lower. No sizeable difference was found between the two compounds as regards their preoestrus-inducing activity.     

Effect of genome size andrrn gene copy numbers of PCR amplification products of 16S rRNA genes from mixed bacterial species

Results and conclusion As determined by image analysis of SYBR green-stained amplification products the experimentally determined ratio corresponded well with the expected ratio calculated from the number ofrrn genes per equimolar amount of DNA in mixtures containing DNA ofEscherichia coli and Thermus thermophilus and DNA ofPseudomonas aeruginosa and T. thermophilus. The values for the pairBacillus subtilis and T. thermophilus showed higher deviation from the predicted value. The dependence of the amount of 16S rDNA amplification products on these two parameters makes it impossible to quantify the number of species present in 16S rDNA clone library of an environmental sample, as long as these two parameters are unknown for these species.     

N-acetylglucosaminyltransferase V modifies the signaling pathway of epidermal growth factor receptor

Transfection of sense cDNA of N-acetylglucosamyltransferase V (GnTV-S) into human H7721 hepatocarcinoma cells resulted in an increase in the N-acetylglucosamine1,6mannose1,3- branch (GnT-V product) on the N-glycans of epidermal growth factor (EGF) receptor (EGFR), and promotion of its EGF binding and tyrosine autophosphorylation, but showed little effect on the expression of EGFR protein. The phosphorylation at T308, S473 and tyrosine residue(s) and the activity of protein kinase B (Akt/PKB) as well as the phosphorylation of p42/44 mitogen-activated protein kinase (MAPK) and MAPK kinase (MEK) before and after EGF stimulation were concomitantly increased. Conversely, in the antisense GnT-V (GnTV-AS)-transfected H7721 cells, all the results were the reverse of those with GnTV-S-transfected cells. After the cells were treated with 1-deoxymannojirimycin, an inhibitor of N-glycan processing at high mannose, or antibody against the extracellular glycan domain of EGFR, the differences in PKB activity, p42/44 MAPK and MEK phosphorylation among GnTV-S-, GnTV-AS- and mock-transfected cells were significantly attenuated. These findings indicate that the altered expression of GnT-V will change the glycan structure and function of EGFR, which may modify downstream signal transduction.Received 24 March 2004; received after revision 1 May 2004; accepted 25 May 2004