129 research outputs found

    Interaction and Modulation of Two Antagonistic Cell Wall Enzymes of Mycobacteria

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    Bacterial cell growth and division require coordinated cell wall hydrolysis and synthesis, allowing for the removal and expansion of cell wall material. Without proper coordination, unchecked hydrolysis can result in cell lysis. How these opposing activities are simultaneously regulated is poorly understood. In Mycobacterium tuberculosis, the resuscitation-promoting factor B (RpfB), a lytic transglycosylase, interacts and synergizes with Rpf-interacting protein A (RipA), an endopeptidase, to hydrolyze peptidoglycan. However, it remains unclear what governs this synergy and how it is coordinated with cell wall synthesis. Here we identify the bifunctional peptidoglycan-synthesizing enzyme, penicillin binding protein 1 (PBP1), as a RipA-interacting protein. PBP1, like RipA, localizes both at the poles and septa of dividing cells. Depletion of the ponA1 gene, encoding PBP1 in M. smegmatis, results in a severe growth defect and abnormally shaped cells, indicating that PBP1 is necessary for viability and cell wall stability. Finally, PBP1 inhibits the synergistic hydrolysis of peptidoglycan by the RipA-RpfB complex in vitro. These data reveal a post-translational mechanism for regulating cell wall hydrolysis and synthesis through protein–protein interactions between enzymes with antagonistic functions

    Kinetic Modeling of the Assembly, Dynamic Steady State, and Contraction of the FtsZ Ring in Prokaryotic Cytokinesis

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    Cytokinesis in prokaryotes involves the assembly of a polymeric ring composed of FtsZ protein monomeric units. The Z ring forms at the division plane and is attached to the membrane. After assembly, it maintains a stable yet dynamic steady state. Once induced, the ring contracts and the membrane constricts. In this work, we present a computational deterministic biochemical model exhibiting this behavior. The model is based on biochemical features of FtsZ known from in vitro studies, and it quantitatively reproduces relevant in vitro data. An essential part of the model is a consideration of interfacial reactions involving the cytosol volume, where monomeric FtsZ is dispersed, and the membrane surface in the cell's mid-zone where the ring is assembled. This approach allows the same chemical model to simulate either in vitro or in vivo conditions by adjusting only two geometrical parameters. The model includes minimal reactions, components, and assumptions, yet is able to reproduce sought-after in vivo behavior, including the rapid assembly of the ring via FtsZ-polymerization, the formation of a dynamic steady state in which GTP hydrolysis leads to the exchange of monomeric subunits between cytoplasm and the ring, and finally the induced contraction of the ring. The model gives a quantitative estimate for coupling between the rate of GTP hydrolysis and of FtsZ subunit turnover between the assembled ring and the cytoplasmic pool as observed. Membrane constriction is chemically driven by the strong tendency of GTP-bound FtsZ to self-assembly. The model suggests a possible mechanism of membrane contraction without a motor protein. The portion of the free energy of GTP hydrolysis released in cyclization is indirectly used in this energetically unfavorable process. The model provides a limit to the mechanistic complexity required to mimic ring behavior, and it highlights the importance of parallel in vitro and in vivo modeling

    The World's Rediscovered Species: Back from the Brink?

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    Each year, numerous species thought to have disappeared are rediscovered. Yet, do these rediscoveries represent the return of viable populations or the delayed extinction of doomed species? We document the number, distribution and conservation status of rediscovered amphibian, bird, and mammal species globally. Over the past 122 years, at least 351 species have been rediscovered, most occurring in the tropics. These species, on average, were missing for 61 years before being rediscovered (range of 3–331 years). The number of rediscoveries per year increased over time and the majority of these rediscoveries represent first documentations since their original description. Most rediscovered species have restricted ranges and small populations, and 92% of amphibians, 86% of birds, and 86% of mammals are highly threatened, independent of how long they were missing or when they were rediscovered. Under the current trends of widespread habitat loss, particularly in the tropics, most rediscovered species remain on the brink of extinction

    Targeting the Wolbachia Cell Division Protein FtsZ as a New Approach for Antifilarial Therapy

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    Filarial nematode parasites are responsible for a number of devastating diseases in humans and animals. These include lymphatic filariasis and onchocerciasis that afflict 150 million people in the tropics and threaten the health of over one billion. The parasites possess intracellular bacteria, Wolbachia, which are needed for worm survival. Clearance of these bacteria with certain antibiotics leads to parasite death. These findings have pioneered the approach of using antibiotics to treat and control filarial infections. In the present study, we have investigated the cell division process in Wolbachia for new drug target discovery. We have identified the essential cell division protein FtsZ, which has a GTPase activity, as an attractive Wolbachia drug target. We describe the molecular characterization and catalytic properties of the enzyme and demonstrate that the GTPase activity is inhibited by the natural product, berberine, and small molecule inhibitors identified from a high-throughput screen. We also found that berberine was effective in reducing motility and reproduction in B. malayi parasites in vitro. Our results should facilitate the discovery of selective inhibitors of FtsZ as a novel antibiotic approach for controlling filarial infection

    Acute Sleep Deprivation and Circadian Misalignment Associated with Transition onto the First Night of Work Impairs Visual Selective Attention

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    Background: Overnight operations pose a challenge because our circadian biology promotes sleepiness and dissipates wakefulness at night. Since the circadian effect on cognitive functions magnifies with increasing sleep pressure, cognitive deficits associated with night work are likely to be most acute with extended wakefulness, such as during the transition from a day shift to night shift. Methodology/Principal Findings: To test this hypothesis we measured selective attention (with visual search), vigilance (with Psychomotor Vigilance Task [PVT]) and alertness (with a visual analog scale) in a shift work simulation protocol, which included four day shifts followed by three night shifts. There was a nocturnal decline in cognitive processes, some of which were most pronounced on the first night shift. The nighttime decrease in visual search sensitivity was most pronounced on the first night compared with subsequent nights (p = .04), and this was accompanied by a trend towards selective attention becoming ‘fast and sloppy’. The nighttime increase in attentional lapses on the PVT was significantly greater on the first night compared to subsequent nights (p<.05) indicating an impaired ability to sustain focus. The nighttime decrease in subjective alertness was also greatest on the first night compared with subsequent nights (p<.05). Conclusions/Significance: These nocturnal deficits in attention and alertness offer some insight into why occupational errors, accidents, and injuries are pronounced during night work compared to day work. Examination of the nighttime vulnerabilities underlying the deployment of attention can be informative for the design of optimal work schedules and the implementation of effective countermeasures for performance deficits during night work

    Zoofolkloristika: Prvi uvidi na putu prema novoj disciplini

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    The author notes that new, more complex researches of connections between animals, nature and connections to humans are needed in Slovenian and European folklore, literature and cultural studies, due to new ecological and ethical findings in the wider social and cultural environment and a changing order of the world, which has moved the focus from anthropocentrism into ecocentrism. The discussion builds upon various theoretical discourses, new concepts and multidisciplinary knowledge, to create the foundations, guidelines and directions for a new academic discipline of zoofolkloristics. Furthermore, new theoretical and analytical discourses should enable zoofolkloristics to provide an insight into changes in human attitudes to animals, in both folklore and within traditional and contemporary ritual practices, or their redefinition, and at the same time exert influence upon legal safety of non-human subjectivities.Autorica polazi od tvrdnje da su potrebna nova, kompleksnija istraživanja veza između životinja, prirode i poveznica s ljudima u slovenskoj i europskoj folkloristici, znanosti o književnosti i kulturnim studijima, zbog novih ekoloških i etičkih uvida u širu društveno-kulturnu okolinu i promjene u svijetu, koje su dovele do promjene od antropocentrizma k ekocentrizmu. U radu se na temelju različitih teorijskih diskursa, novih pojmova i multidisciplinarnog znanja stvaraju temelji, smjernice i pravci istraživanja u novoj znanstvenoj disciplini zoofolkloristike. Autorica tvrdi da bi etnologija, folkloristika i znanost o književnosti mogle imati ključnu ulogu u razvoju ekološke svijesti, znanstvenih terenskih istraživanja i kulturne ekologije te tako omogućiti prelazak iz ekologije u eshatologiju. Rad započinje opisom percepcije životinja i odnosa ljudi i životinja, gdje autorica govori o temeljnim pojmovima koji utječu na uvođenje ove nove znanstvene discipline, kao što su: životinja kao ispitanik, individualni pristup životinji, sinantropski i antropofilni pogled na čovjeka i životinju te antropomorfizam. Zatim se govori o povijesnim i suvremenim filozofsko-antropološkim diskursima o ljudskom pogledu na životinju te o teoriji specizma. Autorica razmatra predmet nove discipline te kaže da je u centru istraživanja životinja u folkloru u najširem mogućem smislu, u svim sferama narodne kulture, što se proučava iz novih gledišta, nove percepcije i recepcije; u pjesmama, pričama, bajkama, basnama, predajama, poslovicama, izrekama, zagonetkama, šalama, narodnom jeziku, kulturnim praksama, narodnom teatru, mitologiji, narodnoj medicini pa čak i u narodnoj glazbi. Nadalje, autorica daje pregled kulturnih istraživanja životinja u Europi i Sloveniji, te pokazuje da su rasprave o životinjama uglavnom objavljivane u 21. stoljeću, što bi moglo označavati da je riječ o prelasku u novu paradigmu ili čak i novu ontologiju. Autorica se bavi metodološkim i teorijskim pitanjima i pravcima u zoofolklorističkim istraživanjima koje klasificira prema kritičkom diskursu. Rad završava pregledom ciljeva zoofolkloristike, za koju tvrdi da ne bi smjela postati znanstvena disciplina koja nema aktivan utjecaj na društvenopolitički prostor u kojem koegzistiraju ljudski i ne-ljudski subjektiviteti

    Brettanomyces bruxellensis yeasts: impact on wine and winemaking

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    Yeasts belonging to the Brettanomyces/Dekkera genus are non-conventional yeasts, which affect winemaking by causing wine spoilage all over the world. This mini-review focuses on recent results concerning the presence of Brettanomyces bruxellensis throughout the wine processing chain. Here, culture-dependent and independent methods to detect this yeast on grapes and at the very early stage of wine production are encompassed. Chemical, physical and biological tools, devised for the prevention and control of such a detrimental species during winemaking are also presented. Finally, the mini-review identifies future research areas relevant to the improvement of wine safety and sensory profiles
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