Damped oscillations in the adaptive response of the iron homeostasis network of E. coli

Living organisms often have to adapt to sudden environmental changes and reach homeostasis. To achieve adaptation, cells deploy motifs such as feedback in their genetic networks, endowing the cellular response with desirable properties. We studied the iron homeostasis network of E. coli, which employs feedback loops to regulate iron usage and uptake, while maintaining intracellular iron at non‐toxic levels. Using fluorescence reporters for iron‐dependent promoters in bulk and microfluidics‐based, single‐cell experiments, we show that E. coli cells exhibit damped oscillations in gene expression, following sudden reductions in external iron levels. The oscillations, lasting for several generations, are independent of position along the cell cycle. Experiments with mutants in network components demonstrate the involvement of iron uptake in the oscillations. Our findings suggest that the response is driven by intracellular iron oscillations large enough to induce nearly full network activation/deactivation. We propose a mathematical model based on a negative feedback loop closed by rapid iron uptake, and including iron usage and storage, which captures the main features of the observed behaviour. Taken together, our results shed light on the control of iron metabolism in bacteria and suggest that the oscillations represent a compromise between the requirements of stability and speed of response.     

Duration of diapause in the stem borers, Busseola fusca and Chilo partellus

The duration of diapause in the stem borers Busseola fusca (Fuller) and Chilo partellus (Swinhoe) was studied in South Africa by collecting diapausing larvae from the field throughout winter (April–August). B. fusca larvae emerged as moth around the middle of October regardless of the date of collection and the length of time they were kept in the laboratory under constant 21 °C. C. partellus larvae collected in April–June emerged in November, those collected in July emerged in October, and those collected in August emerged in September. Regardless of the collection date C. partellus started to emerge from diapause earlier and moth emergence lasted up to twice as long as in B. fusca. Under laboratory conditions at 60% RH both borer larvae lost about 50% of their body mass during diapause. When provided with water B. fusca larvae lost about 30% of their body mass and adults emerged 20 days earlier than when kept dry. C. partellus, on the other hand, lost only 13% of their body weight and emerged 34 days earlier. The differences between the two species are discussed in light of different types of diapause; i.e., obligatory diapause in B. fusca and facultative diapause in C. partellus.     

An improved method for mass rearing Paratheresia claripalpis for biological control of lepidopteran stalk borers

An improved method for mass rearing Paratheresia claripalpis Van der Wulp (Diptera: Tachinidae), for biological control of lepidopteran stalk borers is described and compared to the commonly used method. The improved method was found to be superior by being less labour intensive and by producing 2.25 times more parasites from the same number of hosts.

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Amélioration de la technique d'élevage de masse de Paratheresia claripalpis pour la lutte biologique contre Diatraea saccharalis

Résumé Une technique d'élevage de massa de la tachinaire, P. claripalpis Van der Wulp, est décrite et comparée à la technique classique. Cette technique correspond à une amélioration puisqu'elle demande moins de travail et produit 2.25 fois plus de parasites pour le même nombre d'hôtes.

     

Antibody-mediated targeting of human single-chain class I MHC with covalently linked peptides induces efficient killing of tumor cells by tumor or viral-specific cytotoxic T lymphocytes

Soluble forms of human MHC class I HLA-A2 were produced in which the peptide binding groove was uniformly occupied by a single tumor or viral-derived peptides attached via a covalent flexible peptide linker to the N terminus of a single-chain -2-microglobulin-HLA-A2 heavy chain fusion protein. A tetravalent version of this molecule with various peptides was found to be functional. It could stimulate T cells specifically as well as bind them with high avidity. The covalently linked single chain peptide-HLA-A2 construct was next fused at its C-terminal end to a scFv antibody fragment derived from the variable domains of an anti-IL-2R subunit-specific humanized antibody, anti-Tac. The scFv–MHC fusion was thus encoded by a single gene and produced in E. coli as a single polypeptide chain. Binding studies revealed its ability to decorate Ag-positive human tumor cells with covalent peptide single-chain HLA-A2 (scHLA-A2) molecules in a manner that was entirely dependent upon the specificity of the targeting Antibody fragment. Most importantly, the covalent scHLA-A2 molecule, when bound to the target tumor cells, could induce efficient and specific HLA-A2-restricted, peptide-specific CTL-mediated lysis. These results demonstrate the ability to generate soluble, stable, and functional single-chain HLA-A2 molecules with covalently linked peptides, which when fused to targeting antibodies, potentiate CTL killing. This new approach may open the way for the development of new immunotherapeutic strategies based on antibody targeting of natural cognate MHC ligands and CTL-based cytotoxic mechanisms.Kfir Oved and Avital Lev contributed equally to this work     

Hibernation by the lepidopteran stalk borers, Busseola fusca and Chilo partellus on grain sorghum

The lepidopteran stalk borers, the indigenous Busseola fusca (Fuller) (Noctuidae), and the exotic Chilo partellus (Swinhoe) (Pyralidae), were studied in grain sorghum fields, Sorghum bicolor (L.) Moench (cultivar SSK-52), in the dry season of 1986 (April–October) at Delmas and Brits, Transvaal, South Africa. More than 90% of plants at both sites were infested, but as winter progressed, the proportion of plants infested and the level of infestation dropped gradually. The borer larvae hibernated inside the dry stalks, but the location differed: 65% of B. fusca were in the lower third and 30% in the middle third, whereas for C. partellus it was 45 and 50% respectively. Both borers overwintered as larvae in either of the last three instars, but the proportions of larval instars were different. About 82% of B. fusca hibernated as 6th and 16% as 5th larval instars whereas with C. partellus it was 40 and 45% respectively. C. partellus started to emerge from diapause in the second part of August and it lasted until the first week of November, a period of 12 weeks, but with B. fusca on the other hand pupation lasted only 3 weeks during October–November. Parasitism was low on both species, but higher on B. fusca. Three parasites, Chelonus curvinaculatus Cameron, Chelonus sp. and Pristomerus sp. hibernated in the borers, one parasite Iphiaulax sp., hibernated in its cocoon inside the stalks, and two parasites Apanteles sesamiae Cameron and Bracon sp. were active in winter. Two ants, Pheidole megacephala Febricius, and Dorylus helvolus (L.), preyed on the borer larvae in winter.

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Hibernation des chenilles de deux espèces de mineuses, Busseola fusca et Chilo partellu dans les tiges de Sorghum bicolor

Résumé Deux chenilles mineuses de tiges, B. fusca, Noctuidae indigène, et C. partellus, Pyralidae exotique, ont été étudiées dans des champs de S. bicolor (cultivar SSK-52), pendant la saison sèche de 1986 (Avril–Octobre) à Delmas et Brits, au Transvaal en Afrique du Sud. Plus de 90% des pieds des deux stations étaient infestés, mais plus l'hiver avançait, plus la proportion de plantes in festées et le taux d'infestation diminuaient. Les chenilles ont hiverné dans les tiges sèches, mais leurs positions étaient différentes: 65% des B. fusca étaient dans le tiers inférieur et 30% dans le tiers médian, tandis que pour C. partellus, il s'agissait respectivement de 45 et 50%. Les duex chenilles hivernaient à l'un des 3 derniers stades larvaires, mais les proportions étaient différentes. Pour B. fusca environ 82% hivernaient au 6ème stade et 16% au 5ème stade, tandis que pour C. partellus c'étaient 40 et 45% respectivement. C. partellus a commencé à sortir de diapause pendant la seconde moitié d'Août et a continué jusqu'à la première semaine de Novembre, soit durant une période de 12 semaines, mais avec B. fusca la nymphose a duré seulement 3 semaines en Octobre–Novembre. Le parasitisme des deux espèces a été faible, mais plus important chez B. fusca. Trois parasites, Chelonus curvimaculatus Cameron, Chelonus sp. et Pristomerus sp. ont hiverné dans leurs cocons à l'intérieur des tiges, et deux parasites, Apanteles sesamiae Cameron et Bracon sp. étaient actifs en hiver. Deux fourmis, Pheidole megacephala Fabricius et Dorylus helvolus L. capturaient des chenilles de ces mineuses pendant l'hiver.

     

Application of a fluorescent biosensor based‐on magneto‐γ‐Fe2O3–methyldopa nanoparticles for adsorption of human serum albumin

Understanding and controlling the interaction between the polymer methyldopa (2‐amino‐3‐(3,4‐dihydroxyphenyl)‐2‐methyl‐propanoic acid) (PMDP)–γ‐Fe₂O₃ nanoparticles and biological fluids is important if the potential of nanoparticles (NPs) in biomedicine is to be realized. Physicochemical studies on the interactions between proteins and NPs are influenced by the surface properties of the NPs. To identify the effects of the NP surface, interactions between human serum albumin (HSA) and PMDP–γ‐Fe₂O₃ NPs were investigated. Here, the adsorption of HSA onto small (10–30 nm diameter) PMDP–γ‐Fe₂O₃ NPs was quantitatively analyzed using spectroscopic methods. The fluorescence quenching data were checked for the inner‐filter effect, the main confounding factor in the observed quenching. The binding constants, K_a, were calculated at different temperatures, using a nonlinear fit to the experimental data, and the thermodynamic parameters ?H, ?S and ?G were given. The obtained thermodynamic signature suggests that hydrophobic interactions at least are present. This result indicates that the structure of the protein turns from a structureless denatured state at pH 3 into an ordered biologically active native state on addition of PMDP–γ‐Fe₂O₃ NPs. Copyright © 2015 John Wiley & Sons, Ltd.     

The Response to High CO2 Levels Requires the Neuropeptide Secretion Component HID-1 to Promote Pumping Inhibition

Carbon dioxide (CO₂) is a key molecule in many biological processes; however, mechanisms by which organisms sense and respond to high CO₂ levels remain largely unknown. Here we report that acute CO₂ exposure leads to a rapid cessation in the contraction of the pharynx muscles in Caenorhabditis elegans. To uncover the molecular mechanisms underlying this response, we performed a forward genetic screen and found that hid-1, a key component in neuropeptide signaling, regulates this inhibition in muscle contraction. Surprisingly, we found that this hid-1-mediated pathway is independent of any previously known pathways controlling CO₂ avoidance and oxygen sensing. In addition, animals with mutations in unc-31 and egl-21 (neuropeptide secretion and maturation components) show impaired inhibition of muscle contraction following acute exposure to high CO₂ levels, in further support of our findings. Interestingly, the observed response in the pharynx muscle requires the BAG neurons, which also mediate CO₂ avoidance. This novel hid-1-mediated pathway sheds new light on the physiological effects of high CO₂ levels on animals at the organism-wide level.     

The mutagenicity of natural products from marine algae.

5 polyhalogenated hydrocarbon natural products isolated from the marine red alga Plocamium spp. were tested for mutagenicity in the Ames reversion assay. All 5 of the compounds induced revertants in Salmonella typhimurium strains TA100 and TA1535, indicating the mutational events involved base substitutions. One of the compounds, designated cross-conjugated ketone, was shown to be almost 200 times more effective as a mutagen than was ethyl methanesulfonate.     

Monocyte and macrophage activation by lipoteichoic Acid is independent of alanine and is potentiated by hemoglobin

Lipoteichoic acids (LTAs) are Gram-positive bacterial cell wall components that elicit mononuclear cell cytokine secretion. Cytokine-stimulating activity is thought to be dependent on retaining a high level of ester-linked D-alanine residues along the polyglycerol phosphate backbone. However, Streptococcus pyogenes LTA essentially devoid of D-alanine caused human and mouse cells to secrete as much IL-6 as LTA with a much higher D-alanine content. Furthermore, hemoglobin (Hb) markedly potentiates the stimulatory effect of various LTAs on mouse macrophages or human blood cells, regardless of their d-alanine content. LTA and Hb appear to form a molecular complex, based on the ability of each to affect the other's migration on native acrylamide gels, their comigration on these gels, and the ability of LTA to alter the absorption spectra of Hb. Because S. pyogenes is known to release LTA and secrete at least two potent hemolytic toxins, LTA-Hb interactions could occur during streptococcal infections and might result in a profound alteration of the local inflammatory response.