The effect of antibiotics on the photocycle and protoncycle of purple membrane suspensions

The interrelation was studied between the phototransient absorbing maximally at 412 nm (M₄₁₂) and light-induced proton release under steady-state conditions in aqueous suspensions of ‘purple membrane’ derived from Halobacterium halobium. The decay of M₄₁₂ was slowed down by the simultaneous application of the ionophoric antibiotics valinomycin and beauvericin. The former had only slight activity alone and the latter was effective only in conjunction with valinomycin. The steady-state concentration of M₄₁₂ which was formed on illumination was a direct function of the concentration of valinomycin. Maximum stabilization of M₄₁₂ was obtained when the valinomycin was approximately equimolar with the bacteriorhodopsin. Addition of salts to the medium increased the number of protons released per molecule of M₄₁₂ without affecting the level of M₄₁₂ which was produced by continuous illumination. The effectiveness of the salts in this respect depended on the nature of the cation. Ca²⁺ and their antagonists La³⁺ and ruthenium red were found to have especially high affinity for the system. The extent of light-induced acidification could not be enhanced by increasing the pH of the medium from 6.5 to 7.8. The possible mechanism of action of the ionophores and of the cations on the photocycle and on the proton cycle is discussed.     

Chemical identification of cysteine as palmitoylation site in a transmembrane protein (Semliki Forest virus E1)

The palmitoylation site of the membrane glycoprotein E1 of Semliki Forest virus (SFV) has been identified by chemical analysis of an acylpeptide. 3H-Palmitoylated E1 isolated from SFV grown in baby hamster kidney cells was digested with chymotrypsin and the resulting peptides subjected to high performance liquid chromatography on a wide-pore column. The 3H-acylated peptide fraction peaked at above 60% 2-propanol in the eluent, indicating its hydrophobic character. Polyacrylamide gel electrophoresis analysis revealed a molecular weight of about Mr = 6000 for the radiolabeled peptide. Manual sequencing of this material by the 4-N,N'-dimethylaminoazobenzene-4'-isothiocyanate/phenylisothiocyanate procedure on solid phase revealed the amino-terminal sequence Ala-Ala-Ser-His-Ser-Asn-Val-Val-Phe-Pro. The same peptide also labels with [35S]cysteine. Comparison with the deduced amino acid sequence of E1 revealed that the palmitoylated peptide contains at least 43 amino acid residues, and thus includes the membrane spanning region down to the only cysteine residue five positions up from the carboxyl terminus of E1. Since [3H]palmitic acid was cleaved from E1 with thiol reagents, and since the peptide labels with [14C]iodoacetamide only after the release of fatty acids by hydroxylamine treatment, cysteine in position 433 represents the palmitoylation site in SFV E1.     

High level of divergence of male-reproductive-tract proteins, between Drosophila melanogaster and its sibling species, D. simulans

We compared male-reproductive-tract polypeptides of Drosophila melanogaster and D. simulans by using two-dimensional gel electrophoresis. Approximately 64% of male-reproductive-tract polypeptides were identical between two randomly chosen isofemale lines from these two species, compared with 83% identity for third-instar imaginal wing-disc polypeptides. Qualitatively similar differences were found between reproductive tracts and imaginal discs when D. sechellia was compared with D. melanogaster and with D. simulans. When genic polymorphism was taken into account, approximately 10% of male- reproductive-tract polypeptides were apparently fixed for different alleles between D. melanogaster and D. simulans; this proportion is the same as that found for soluble enzymes by one-dimensional gel electrophoresis. Strikingly, approximately 20% of male-reproductive- tract polypeptides of either D. melanogaster or D. simulans had no detectable homologue in the other species. We propose that proteins of the Drosophila male reproductive tract may have diverged more extensively between species than have other types of proteins and that much of this divergence may involve large changes in levels of polypeptide expression.      

The temporal pattern of interphase prolongation and nuclear activities during early embryogenesis in teleostei

Summary Temporal relationship between the beginning of the increase of interphase duration, RNA synthesis in the nuclei and morphogenetic function of nuclei have been studied using dimensionless criteria of development duration. Experiments were performed in the trout (Salmo trutta m.fario andSalmo irideus), the whitefish (Coregonus lavaretus), the pike (Esox lucius) and the carp (Cyprinus caprio). In all these species interphase begins to lengthen approximately at the same developmental stages. The onset of morphogenetic nuclear function in the pike and whitefish coincides with the onset of interphase prolongation; in the carp RNA synthesis in the nuclei and morphogenetic nuclear function were revealed by 2 ₀ earlier than onset of interphase prolongation, and in the trout, by 5 and 13 ₀ later, respectively. Thus, a temporal disconnection between the onset of interphase prolongation, RNA synthesis in the nuclei and morphogenetic nuclear function was established for the first time. It can be suggested that the character of temporal relationship between these processes depends on the relative quantity of yolk in the eggs.

---

Zusammenfassung In der Embryogenese von Forelle, Weißfisch, Hecht und Karpfen ist untersucht worden, welche zeitlichen Beziehungen bestehen zwischen dem Beginn der Zunahme der Interphasedauer, der RNS-Synthese in den Kernen und der morphogenetischen Funktion der Kerne. Bei allen genannten Arten verlängert sich die Interphase etwa in dem gleichen Entwicklungsstadium. Beim Hecht und beim Weißfisch beginnt die morphogenetische Funktion der Kerne gleichzeitig mit der Verlängerung der Interphasedauer; beim Karpfen setzen RNS-Synthese und morphogenetische Funktion der Kerne zwei Zeiteinheiten ( ₀) früher ein als der Beginn der Interphaseverlängerung, bei der Forelle dagegen 5 bzw. 13 Zeiteinheiten später. Damit ist erstmals gezeigt worden, daß kein zeitlicher Zusammenhang besteht zwischen dem Beginn von Interphaseverlängerung, RNS-Synthese in den Kernen und morphogenetischer Funktion der Kerne. Vermutlich hängt die zeitliche Beziehung zwischen diesen Vorgängen vom relativen Dottergehalt der Eier ab.

     

Latent autoantibodies to cardiolipin in the blood serum of healthy subjects

Pooled health blood donors' sera were fractionated by gel filtration and/or ion exchange chromatography on strong anion-exchanger. Measurement of anticardiolipin antibodies levels by ELISA method shows that gel filtration at pH 4.05 and 9.2 (complex-degradation conditions) leads to increase in summary levels while after elution at neutral pH such an effect did not appear. Light increasing of anticardiolipin levels was also noted after fractionation on QAE-Sephadex. Data obtained state that anticardiolipin autoantibodies amount in sera is greater than it can be detected by direct measurement in untreated serum samples. Existence of "hidden" anticardiolipin autoantibodies supposes the hypothesis about alternative way of humoral immunity regulation by blocking anti-self antigens activities with serum biopolymers.     

Improving the Control of Tetranychus urticae on Edible Glasshouse Crops Using a Specialist Coccinellid ( Stethorus punctillum Weise) and a Generalist Mite ( Amblyseius californicus McGregor) as Biocontrol Agents

Current glasshouse biological control practice relies on regular prophylactic introductions of one or two 'best' species of natural enemy. Whilst this is effective for much of the time, occasional failures occur due to factors such as differences in response to seasonal changes in environmental conditions and/or host plant effects. This study looks at the predatory behaviour of a specialist coccinellid, Stethorus punctillum Weise, and a generalist mite, Amblyseius californicus McGregor (which predate on the two-spotted spider mite, Tetranychus urticae ) in order to assess how they responded to temperatures and relative humidities typical of glasshouse conditions on four edible crop plant species. Activity (distance covered, time spent walking, walking speed, angular velocity, and turning rate) was recorded at 20, 25 and 30 o C and at relative humidity (RH) levels of 33, 65 and 90%, on tomato, pepper, aubergine and cucumber leaves, and analysed using video-computer techniques. The results show that the activity of S. punctillum significantly increased at higher temperature levels. Host plant species also strongly influenced the performance of the predator, with it being most active on pepper and tomato and least active on aubergines. RH had no significant influence. The activity and predation by A. californicus increased at low humidity levels, especially in terms of time spent moving and number of prey killed. Temperature levels had no significant influence, but host plant species strongly influenced the performance of the predator, which was most active on pepper, and least active on aubergines. Further research was conducted with semi-field trials to investigate the efficacy in controlling TSSM with different combination of predators. When contrasting the commercially available predatory mite Phytoseiulus persimilis , used alone, compared with its use in a treatment with a combination of predator species, there was a stronger decrease in TSSM numbers on the crop plants in the latter treatment.     

Absence of intrinsic electric conductivity in single dsDNA molecules

The intrinsic dc conductivity of long, individual lambda phage dsDNA molecules has been investigated by ultrasensitive low current-voltage-spectroscopy (IV) under ambient conditions and controlled low humidity inert gas atmosphere on microfabricated metal-insulator-metal gap structures. We found a strong dependence of the measured conductivity on the apparent humidity, which we attribute to capillary condensation of water to the immobilized DNA molecules, giving rise to additional ionic currents. Additional IV-spectroscopy experiments under controlled argon atmosphere always revealed a significant drop in electrical conductivity to 4 x 10(-15)AV(-1)microm(-1), indicating almost no considerable contribution of electrical long range charge transport.     

Organ tropism of Sendai virus in mice: proteolytic activation of the fusion glycoprotein in mouse organs and budding site at the bronchial epithelium.

Wild-type Sendai virus is exclusively pneumotropic in mice, while a host range mutant, F1-R, is pantropic. The latter was attributed to structural changes in the fusion (F) glycoprotein, which was cleaved by ubiquitous proteases present in many organs (M. Tashiro, E. Pritzer, M. A. Khoshnan, M. Yamakawa, K. Kuroda, H.-D. Klenk, R. Rott, and J. T. Seto, Virology 165:577-583, 1988). These studies were extended by investigating, by use of an organ block culture system of mice, whether differences exist in the susceptibility of the lung and the other organs to the viruses and in proteolytic activation of the F protein of the viruses. Block cultures of mouse organs were shown to synthesize the viral polypeptides and to support productive infections by the viruses. These findings ruled out the possibility that pneumotropism of wild-type virus results because only the respiratory organs are susceptible to the virus. Progeny virus of F1-R was produced in the activated form as shown by infectivity assays and proteolytic cleavage of the F protein in the infected organ cultures. On the other hand, much of wild-type virus produced in cultures of organs other than lung remained nonactivated. The findings indicate that the F protein of wild-type virus was poorly activated by ubiquitous proteases which efficiently activated the F protein of F1-R. Thus, the activating protease for wild-type F protein is present only in the respiratory organs. These results, taken together with a comparison of the predicted amino acid substitutions between the viruses, strongly suggest that the different efficiencies among mouse organs in the proteolytic activation of F protein must be the primary determinant for organ tropism of Sendai virus. Additionally, immunoelectron microscopic examination of the mouse bronchus indicated that the budding site of wild-type virus was restricted to the apical domain of the epithelium, whereas budding by F1-R occurred at the apical and basal domains. Bipolar budding was also observed in MDCK monolayers infected with F1-R. The differential budding site at the primary target of infection may be an additional determinant for organ tropism of Sendai virus in mice.     

Inhibition of the Multiplication of Vesicular Stomatitis and Newcastle Disease Virus by 2-Deoxy-d-Glucose

The production of infectious vesicular stomatitis (VSV) and Newcastle disease virus can be completely inhibited by 2-deoxy-d-glucose in pyruvate-containing medium, if virus either grown in pyruvate-containing medium or dialyzed against phosphate-buffered saline is used for infection. Under these conditions, the synthesis of all VSV proteins is reduced. VSV RNA, which is synthesized at reduced rates, seems to be unstable. The effect is completely reversible. If virus grown in glucose-containing medium is used for infection, the production of both viruses is not significantly inhibited by 2-deoxy-d-glucose. Under these conditions the production of the VSV glycoprotein is specifically impaired, but does not lead to a marked reduction of the yield of infectious virus.     

In vitro reactions of sugarcane (Saccharum SP.) plantlets inoculated with 2 strains of Xanthomonas albilineans (Ashby) Dowson

The symptoms of the leaf scald disease can be reproduced in vitro through the inoculation of sugarcane tissue culture plantlets. The pathogen is detected in the inoculated plantlet and is maintained at the surface of the base of the plantlets grown in vitro. Two strains of X. albilineans belonging to different serovars and lysovars reacted like pathotypes. The importance of the plant incubation temperature is clearly demonstrated. Further, in vitro the disease goes through the same phase of latency as in the field.