Combination drugs, an emerging option for antibacterial therapy

The emerging and sustained resistance to antibiotics and the poor pipeline of new antibacterials is creating a major health issue worldwide. Bacterial pathogens are increasingly becoming resistant even to the most recently approved antibiotics. Few antibiotics are being approved by regulatory organizations, which reflects both the difficulty of developing such agents and the fact that antibiotic discovery programs have been terminated at several major pharmaceutical companies in the past decade. As a result, the output of the drug pipelines is simply not well positioned to control the growing army of resistant pathogens, although academic institutions and smaller companies are trying to fill that gap. An emerging option to fight such pathogens is combination therapy. Combinations of two antibiotics or antibiotics with adjuvants are emerging as a promising therapeutic approach. This article provides and discusses clinical and scientific challenges to support the development of combination therapy to treat bacterial infections.     

Chaperone Hsp27, a novel subunit of AUF1 protein complexes, functions in AU-rich element-mediated mRNA decay

Controlled, transient cytokine production by monocytes depends heavily upon rapid mRNA degradation, conferred by 3' untranslated region-localized AU-rich elements (AREs) that associate with RNA-binding proteins. The ARE-binding protein AUF1 forms a complex with cap-dependent translation initiation factors and heat shock proteins to attract the mRNA degradation machinery. We refer to this protein assembly as the AUF1- and signal transduction-regulated complex, ASTRC. Rapid degradation of ARE-bearing mRNAs (ARE-mRNAs) requires ubiquitination of AUF1 and its destruction by proteasomes. Activation of monocytes by adhesion to capillary endothelium at sites of tissue damage and subsequent proinflammatory cytokine induction are prominent features of inflammation, and ARE-mRNA stabilization plays a critical role in the induction process. Here, we demonstrate activation-induced subunit rearrangements within ASTRC and identify chaperone Hsp27 as a novel subunit that is itself an ARE-binding protein essential for rapid ARE-mRNA degradation. As Hsp27 has well-characterized roles in protein ubiquitination as well as in adhesion-induced cytoskeletal remodeling and cell motility, its association with ASTRC may provide a sensing mechanism to couple proinflammatory cytokine induction with monocyte adhesion and motility.     

Protein interaction mapping on a functional shotgun sequence of Rickettsia sibirica

Protein interaction maps can reveal novel pathways and functional complexes, allowing ‘guilt by association’ annotation of uncharacterized proteins. To address the need for large-scale protein interaction analyses, a bacterial two-hybrid system was coupled with a whole genome shotgun sequencing approach for microbial genome analysis. We report the first large-scale proteomics study using this system, integrating de novo genome sequencing with functional interaction mapping and annotation in a high-throughput format. We apply the approach by shotgun sequencing and annotating the genome of Rickettsia sibirica strain 246, an obligate intracellular human pathogen among the Spotted Fever Group rickettsiae. The bacteria invade endothelial cells and cause lysis after large amounts of progeny have accumulated. Little is known about specific Rickettsial virulence factors and their mode of pathogenicity. Analysis of the combined genomic sequence and protein–protein interaction data for a set of virulence related Type IV secretion system (T4SS) proteins revealed over 250 interactions and will provide insight into the mechanism of Rickettsial pathogenicity.     

Application of wide-spectrum light-emitting diodes in micropropagation of popular ornamental plant species: a study on plant quality and cost reduction

In the present study, the applicability of four wide-spectrum light-emitting diodes (LEDs) emitting warm light (AP67, AP673L, G2, and NS1) was determined for the micropropagation of five popular ornamental plant species: Chrysanthemum × grandiflorum, Gerbera jamesonii, Heuchera × hybrida, Ficus benjamina, and Lamprocapnos spectabilis. Plantlets were grown in a growth room with a 16-h photoperiod. The photosynthetic photon flux density was set at 62–65 μmol m⁻² s⁻¹. The composition of the media and subculture timing were adjusted to the needs of each species tested. The results were compared to the cool daylight-emitting fluorescent (FL) control (TLD 36W/54). In most of the species studied (except for F. benjamina), the highest propagation ratios, or ratios similar to the FL control, were observed under the red- and far-red-abundant G2 LEDs. NS1 spectrum (with the highest proportion of blue and green light) was also efficient for G. jamesonii and L. spectabilis, and it provided a similar propagation ratio as the FL control. Light quality affected shoot length, number of leaves, callus regeneration, and the biosynthesis of chlorophyll. This influence, however, was species-dependent. Lighting conditions did not affect the dry matter and rooting in most of the species tested, except for G. jamesonii. The substitution of FLs with G2 LEDs can result in a 50% reduction of annual electricity costs, while the application of NS1 lamps can generate savings of up to 75%. In conclusion, the G2 LED lighting system seemed to be the most suitable in terms of propagation efficiency, plantlet quality, and cost reduction.

     

Reconstructing the palaeoenvironment of the Middle Russian Sea during the Middle–Late Jurassic transition using stable isotope ratios of cephalopod shells and variations in faunal assemblages

Oxygen and carbon isotope data of well-preserved belemnite rostra and ammonite shells are presented from the Callovian–Oxfordian boundary (uppermost Lamberti to lowermost Cordatum zones) of the Dubki section near Saratov in the Russian Platform. Palaeotemperatures calculated for nektobenthic belemnites (averages of 5 °C and 8 °C for cylindroteuthids and belemnopseids, respectively) show the presence of cold bottom waters in the central part of the Middle Russian Sea during the studied interval. Palaeotemperatures calculated for ammonites, which are assumed to have lived in near-surface waters, are considerably higher (average 13 °C). The presented data show a vertical thermal gradient in the Middle Russian Sea. The belemnite oxygen isotope record and the relative abundances of ammonite families in the Dubki section do not correlate with each other probably as a result of different depth habitats of ammonites and belemnites. A review of literature isotope data shows the climatic zonation in European seas at the Middle–Late Jurassic transition. Despite the flux of cold polar waters to the Middle Russian Sea and the area of Scotland there is no evidence for glaciation at the Middle–Late Jurassic transition. Changes in water circulation during a sea-level highstand were likely a source of spreads of cold bottom waters and cardioceratid ammonite fauna in this time period.The belemnite isotope record of the Callovian–Oxfordian boundary in the Russian Platform is characterized by significant scatter of δ¹³C values. No temporal carbon isotope trend is observed. The δ¹³C values of Russian belemnite rostra average 2.6‰ VPDB being 1 to 2‰ higher than the values of coeval Lower Oxfordian belemnites from the area of the Submediterranean ammonite province. Higher (than Submediterranean) δ¹³C values of Russian belemnite rostra are likely related to high biologic productivity and/or high organic matter burial in semi-isolated Boreal–Subboreal marine basins.     

An assessment of sperm motility estimation for evaluation in rams

There is disagreement about the value of estimating spermatozoal motility as a predictor of fertility in rams. We tested the hypothesis that sperm motility can help one to predict lambing rate by evaluating sperm motility in rams prior to breeding and by subsequently breeding the rams regardless of their sperm motility results. A total of 943 rams (Merino, n=630 and Romney Marsh, n=313) was used during two breeding seasons. For fertility evaluation only 804 rams that bred at least ten ewes each were included in the study. Rams were grouped into 4 groups according to sperm motility scores, ranging from the lowest, which included rams with azoospermia, oligospermia, non-motile and single-motile sperm, through rams with 10 to 30%, 30 to 60%, to the highest at 60 to 100% motile sperm. The corresponding fertility rates found in our study in Merino and Romney breeds were 58.3 and 47.4%, 75.1 and 59.1%, 79.4 and 64.1% and 81.5 and 64.0%, respectively. We concluded that there was a statistically confirmed difference in fertility between the lowest and all the other motility categories. The differences in fertility among rams that had motility categories above 10% were not statistically significant. Evaluation of breeding soundness in large scale surveys of rams, based on sperm motility estimation, may be misleading. Clinical examination of genital organs seems to offer a reliable source of information about rams, and may be considered adequate in sheep breeding.