Streptocollin,a Type IV Lanthipeptide Produced by Streptomyces collinus Tü 365

Lanthipeptides are ribosomally synthesized and post‐translationally modified microbial secondary metabolites. Here, we report the identification and isolation of streptocollin from Streptomyces collinus Tü 365, a new member of class IV lanthipeptides. Insertion of the constitutive ermE* promoter upstream of the lanthipeptide synthetase gene stcL resulted in peptide production. The streptocollin gene cluster was heterologously expressed in S. coelicolor M1146 and M1152 with 3.5‐ and 5.5‐fold increased yields, respectively. The structure and ring topology of streptocollin were determined by high resolution MS/MS analysis. Streptocollin contains four macrocyclic rings, with one lanthionine and three methyllanthionine residues. To the best of our knowledge, this is the first report on the isolation of a class IV lanthipeptide in preparative amounts, and on the successful heterologous expression of a class IV lanthipeptide gene cluster.     

Schizosaccharomyces pombe Pmf1p is structurally and functionally related to Mmf1p of Saccharomyces cerevisiae

A novel family of small proteins, termed p14.5 or YERO57c/YJGFc, has been identified. Independent studies indicate that p14.5 family members are multifunctional proteins involved in several pathways, e.g. regulation of translation or activation of the protease mu-calpain. We have previously shown that Mmf1p, a p14.5 of the budding yeast Saccharomyces cerevisiae, is localized in the mitochondria and influences mitochondrial DNA stability. In addition, we have demonstrated that Mmf1p is functionally related to p14.5 of mammalian cells. To explore further the evolutionary conservation of the mitochondrial function(s) of the p14.5s we have extended our study to the fission yeast, Schizosaccharomyces pombe. In this organism two p14.5 homologous proteins are present: Pmf1p (pombe mitochondrial factor 1) and Hpm1p (homologous Pmf1p factor 1). We have generated a specific Pmf1p antibody, which recognizes a single band of approximately 15 kDa in total cellular extracts. Cellular fractionation experiments indicate that Pmf1p localizes in the mitochondria as well as in the cytoplasm. We also show that Pmf1p shares several properties of S. cerevisiae Mmf1p. Indeed, Pmf1p restores the wild-type phenotype when expressed in delta mmf1 S. cerevisiae cells. Deletion of the leader sequence of Pmf1p abrogates its ability to localize in mitochondria and to functionally replace Mmf1p. Thus, these data together with our previous study show that the mitochondrial function(s) of the p14.5 family members are highly conserved in eukaryotic cells.     

Hearing only one side of normal and mobile phone conversations

Mobile (cell) phone conversations are commonly perceived as annoying when conducted in a public space. An experiment is described that demonstrates one factor contributing to this phenomenon: hearing only one side of a conversation makes it more noticeable and intrusive. Two actors repeatedly staged the same conversation under three conditions: cell phone; normal, co-present both audible, and co-present only one audible. After the staged conversation, which took place on a train, a third person obtained verbal ratings from members of the travelling public. As in a previous experiment published in this journal, the cell phone conversation was rated as more noticeable and intrusive than the normal co-present both audible conversation. Critically, a new experimental condition, co-present one-audible, in which both actors were present but only one side of the conversation was heard, produced ratings equivalent to the cell phone condition. This 'need-to-listen' effect is discussed with regard to implications for design and theories of language use.     

Branched Poly(ε-caprolactone)-Based Copolyesters of Different Architectures and Their Use in the Preparation of Anticancer Drug-Loaded Nanoparticles

Limitations associated with the use of linear biodegradable polyesters in the preparation of anticancer nano-based drug delivery systems (nanoDDS) have turned scientific attention to the utilization of branched-chain (co-)polymers. In this context, the present study evaluates the use of novel branched poly(ε-caprolactone) (PCL)-based copolymers of different architectures for the preparation of anticancer nanoparticle (NP)-based formulations, using paclitaxel (PTX) as a model drug. Specifically, three PCL-polyol branched polyesters, namely, a three-arm copolymer based on glycerol (PCL-GLY), a four-arm copolymer based on pentaerythritol (PCL-PE), and a five-arm copolymer based on xylitol (PCL-XYL), were synthesized via ring-opening polymerization and characterized by proton nuclear magnetic resonance (¹H-NMR), gel permeation chromatography (GPC), intrinsic viscosity, differential scanning calorimetry (DSC), X-ray diffraction (XRD), and Fourier-transform infrared (FT-IR) spectroscopy and cytotoxicity. Then, PTX-loaded NPs were prepared by an oil-in-water emulsion. The size of the obtained NPs varied from 200 to 300 nm, while the drug was dispersed in crystalline form in all formulations. High encapsulation efficiency and high yields were obtained in all cases, while FTIR analysis showed no molecular drug polymer. Finally, in vitro drug release studies showed that the studied nanocarriers significantly enhanced the dissolution rate and extent of the drug.     

Controlled release of allyl isothiocyanate from brown algae Laminaria japonica and mesoporous silica MCM-41 for inhibiting food-borne bacteria

Allyl isothiocyanate (AITC), a naturally occurring antimicrobial compound, is an effective inhibitor of various pathogens, but its use in the food industry is limited by its volatility and pungency. The objective of this study was to overcome the volatility of AITC using dried Laminaria japonica and mesoporous silica MCM-41 as its carrier. AITC-loaded L. japonica (raw and deoiled) powder and silica MCM-41 was achieved via vapor adsorption. The study of AITC adsorption and desorption was determined by monitoring sample weight changing with time. AITC presence in L. japonica and MCM-41 samples was confirmed by FTIR spectroscopy. Antimicrobial tests were made against 4 microorganisms: Bacillus cereus, Staphylococcus aureus, Escherichia coli, and Salmonella Typhimurium. Controlled release and antimicrobial activity from MCM-41 was always superior to those from raw and deoiled L. japonica.     

A Regulator Based “Semi-Targeted” Approach to Activate Silent Biosynthetic Gene Clusters

By culturing microorganisms under standard laboratory conditions, most biosynthetic gene clusters (BGCs) are not expressed, and thus, the products are not produced. To explore this biosynthetic potential, we developed a novel “semi-targeted” approach focusing on activating “silent” BGCs by concurrently introducing a group of regulator genes into streptomycetes of the Tübingen strain collection. We constructed integrative plasmids containing two classes of regulatory genes under the control of the constitutive promoter ermE*p (cluster situated regulators (CSR) and Streptomyces antibiotic regulatory proteins (SARPs)). These plasmids were introduced into Streptomyces sp. TÜ17, Streptomyces sp. TÜ10 and Streptomyces sp. TÜ102. Introduction of the CSRs-plasmid into strain S. sp. TÜ17 activated the production of mayamycin A. By using the individual regulator genes, we proved that Aur1P, was responsible for the activation. In strain S. sp. TÜ102, the introduction of the SARP-plasmid triggered the production of a chartreusin-like compound. Insertion of the CSRs-plasmid into strain S. sp. TÜ10 resulted in activating the warkmycin-BGC. In both recombinants, activation of the BGCs was only possible through the simultaneous expression of aur1PR3 and griR in S. sp. TÜ102 and aur1P and pntR in of S. sp. TÜ10.     

Immune Prophylaxis and Therapy for Human Cytomegalovirus Infection

Human Cytomegalovirus (HCMV) infection is widespread and can result in severe sequelae in susceptible populations. Primary HCMV infection of naïve individuals results in life-long latency characterized by frequent and sporadic reactivations. HCMV infection elicits a robust antibody response, including neutralizing antibodies that can block the infection of susceptible cells in vitro and in vivo. Thus, antibody products and vaccines hold great promise for the prevention and treatment of HCMV, but to date, most attempts to demonstrate their safety and efficacy in clinical trials have been unsuccessful. In this review we summarize publicly available data on these products and highlight new developments and approaches that could assist in successful translation of HCMV immunotherapies.     

Continuous-Wave and Passively Mode-Locked Operation of a Cunyite (Cr(4+):Ca(2)GeO(4)) Laser

Continuous-wave and mode-locked Cr(4+):Ca(2)GeO(4) lasers that use a fiber laser pump source were demonstrated. The continuous-wave Cr(4+):Ca(2)GeO(4) laser yielded a maximum output power of 415 mW at 1420 nm and a tuning range of 1335-1492 nm. With a saturable-absorber mirror, 60-ps pulses and 110-mW maximum output power were generated from a passively mode-locked Cr(4+):Ca(2)GeO(4) laser.