Cyclic Amp-Dependent Resuscitation of Dormant Mycobacteria by Exogenous Free Fatty Acids

One third of the world population carries a latent tuberculosis (TB) infection, which may reactivate leading to active disease. Although TB latency has been known for many years it remains poorly understood. In particular, substances of host origin, which may induce the resuscitation of dormant mycobacteria, have not yet been described. In vitro models of dormant (“non-culturable”) cells of Mycobacterium smegmatis (mc(2)155) and Mycobacterium tuberculosis H37Rv were used. We found that the resuscitation of dormant M. smegmatis and M. tuberculosis cells in liquid medium was stimulated by adding free unsaturated fatty acids (FA), including arachidonic acid, at concentrations of 1.6–10 µM. FA addition enhanced cAMP levels in reactivating M. smegmatis cells and exogenously added cAMP (3–10 mM) or dibutyryl-cAMP (0.5–1 mM) substituted for FA, causing resuscitation of M. smegmatis and M. tuberculosis dormant cells. A M. smegmatis null-mutant lacking MSMEG_4279, which encodes a FA-activated adenylyl cyclase (AC), could not be resuscitated by FA but it was resuscitated by cAMP. M. smegmatis and M. tuberculosis cells hyper-expressing AC were unable to form non-culturable cells and a specific inhibitor of AC (8-bromo-cAMP) prevented FA-dependent resuscitation. RT-PCR analysis revealed that rpfA (coding for resuscitation promoting factor A) is up-regulated in M. smegmatis in the beginning of exponential growth following the cAMP increase in lag phase caused by FA-induced cell activation. A specific Rpf inhibitor (4-benzoyl-2-nitrophenylthiocyanate) suppressed FA-induced resuscitation. We propose a novel pathway for the resuscitation of dormant mycobacteria involving the activation of adenylyl cyclase MSMEG_4279 by FAs resulted in activation of cellular metabolism followed later by increase of RpfA activity which stimulates cell multiplication in exponential phase. The study reveals a probable role for lipids of host origin in the resuscitation of dormant mycobacteria, which may function during the reactivation of latent TB

National Projects in the Arctic Zone of the Russian Federation

In 2021, the Russian Federation will assume the chairmanship of the Arctic Council. This fact confirms the special attention of the country’s leadership to the unique geostrategic region. In December 2020, President of the Russian Federation Vladimir Putin, at an online meeting with volunteers and finalists of the contest “Volunteer of Russia”, noted that Russia would grow with the Arctic and the territories of the North. Many Russian citizens’ lives and the state’s success in the international arena depend on this territory’s development. One of the most important mechanisms for achieving the stated goals is national projects (NP), which should make a significant contribution to the development of the country’s territory. The purpose of the study is to assess NP in the subjects that are entirely related to the Arctic zone: Murmansk oblast, Nenets Autonomous Okrug (NAO), Yamalo-Nenets Autonomous Okrug (YANAO), and Chukotka Autonomous Okrug (CHAO). In the course of the work, general scientific research methods were applied: comparative analysis, data comparison, induction, deduction, etc. The study results showed that, despite all the differences in economic and geographical position, financial condition, population, and regional development priorities, all Arctic regions have similar problems that hinder the effective development of territories. Without solving the problems indicated in the work, it is impossible to expect a qualitative «break-through» and implement the NP’s goals

Finding of the Low Molecular Weight Inhibitors of Resuscitation Promoting Factor Enzymatic and Resuscitation Activity

Background: Resuscitation promoting factors (RPF) are secreted proteins involved in reactivation of dormant actinobacteria, including Mycobacterium tuberculosis. They have been considered as prospective targets for the development of new antituberculosis drugs preventing reactivation of dormant tubercle bacilli, generally associated with latent tuberculosis. However, no inhibitors of Rpf activity have been reported so far. The goal of this study was to find low molecular weight compounds inhibiting the enzymatic and biological activities of Rpfs. Methodology/Principal Findings: Here we describe a novel class of 2-nitrophenylthiocyanates (NPT) compounds that inhibit muralytic activity of Rpfs with IC50 1–7 mg/ml. Fluorescence studies revealed interaction of active NPTs with the internal regions of the Rpf molecule. Candidate inhibitors of Rpf enzymatic activity showed a bacteriostatic effect on growth of Micrococcus luteus (in which Rpf is essential for growth protein) at concentrations close to IC50. The candidate compounds suppressed resuscitation of dormant (‘‘non-culturable’’) cells of M. smegmatis at 1 mg/ml or delayed resuscitation of dormant M. tuberculosis obtained in laboratory conditions at 10 mg/ml. However, they did not inhibit growth of active mycobacteria under these concentrations. Conclusions/Significance: NPT are the first example of low molecular weight compounds that inhibit the enzymatic an

25th annual computational neuroscience meeting: CNS-2016

The same neuron may play different functional roles in the neural circuits to which it belongs. For example, neurons in the Tritonia pedal ganglia may participate in variable phases of the swim motor rhythms [1]. While such neuronal functional variability is likely to play a major role the delivery of the functionality of neural systems, it is difficult to study it in most nervous systems. We work on the pyloric rhythm network of the crustacean stomatogastric ganglion (STG) [2]. Typically network models of the STG treat neurons of the same functional type as a single model neuron (e.g. PD neurons), assuming the same conductance parameters for these neurons and implying their synchronous firing [3, 4]. However, simultaneous recording of PD neurons shows differences between the timings of spikes of these neurons. This may indicate functional variability of these neurons. Here we modelled separately the two PD neurons of the STG in a multi-neuron model of the pyloric network. Our neuron models comply with known correlations between conductance parameters of ionic currents. Our results reproduce the experimental finding of increasing spike time distance between spikes originating from the two model PD neurons during their synchronised burst phase. The PD neuron with the larger calcium conductance generates its spikes before the other PD neuron. Larger potassium conductance values in the follower neuron imply longer delays between spikes, see Fig. 17.Neuromodulators change the conductance parameters of neurons and maintain the ratios of these parameters [5]. Our results show that such changes may shift the individual contribution of two PD neurons to the PD-phase of the pyloric rhythm altering their functionality within this rhythm. Our work paves the way towards an accessible experimental and computational framework for the analysis of the mechanisms and impact of functional variability of neurons within the neural circuits to which they belong

26th Annual Computational Neuroscience Meeting (CNS*2017): Part 3 - Meeting Abstracts - Antwerp, Belgium. 15–20 July 2017

This work was produced as part of the activities of FAPESP Research,\ud Disseminations and Innovation Center for Neuromathematics (grant\ud 2013/07699-0, S. Paulo Research Foundation). NLK is supported by a\ud FAPESP postdoctoral fellowship (grant 2016/03855-5). ACR is partially\ud supported by a CNPq fellowship (grant 306251/2014-0)

Reservoirs - Brno, Zluty kopec (Yellow hill)

Diplomová práce pojednává o tvorbě návrhu novostavby centra současného umění a architektury a využití stávajících vodojemů na Žlutém kopci v Brně s možností změny jejich původní funkce. Práce se zabývá možnostmi zpracovaní původního terénu a návazností navrhovaných novostaveb na existující vodojemy. Architektonicky zkoumá možnosti navrhování novostaveb kontextuálně se současným architektonickým výrazem okolních budov a architekturou minulou. Současná situace je taková, že tak nádherná architektonicko-inženýrská památka, jako jsou vodojemy na Žlutém kopci, je prakticky zavřená před očima obyčejných lidí. Pouze jeden ze tří vodojemů je otevřen k návštěvě – nejstarší cihlový vodojem postavený v roce 1872. Hlavním cílem mé práce proto bylo zpřístupnit areál vodojemů veřejnosti a dát lidem možnost navštívit všechny tři vodojemy. K tomu bylo nutné navrhnout vstupní pavilony a bezpečné vstupy do vodojemů a také naplnit vodojemy novou funkcí využití, protože činnost vodojemů byla ukončena v roce 1997, kdy byly odpojeny od vodovodní sítě.The diploma thesis deals with the creation of a design for a new building of the center of contemporary art and architecture and the use of existing reservoirs on Yellow Hill in Brno with the possibility of changing their original function. The work deals with the possibilities of processing the original terrain and the connection of the proposed new buildings to existing reservoirs. Architecturally, it explores the possibilities of designing new buildings contextually with the current architectural expression of the surrounding buildings and past architecture. The current situation is that a beautiful architectural and engineering monument, such as the reservoirs on Yellow Hill, is practically closed to the eyes of ordinary people. Only one of the three reservoirs is open to visit - the oldest brick reservoir built in 1872. The main goal of my work was therefore to make the reservoir area accessible to the public and to give people the opportunity to visit all three reservoirs. To this end, it was necessary to design entrance pavilions and safe entrances to the reservoirs and to fill the reservoirs with a new function, because the reservoirs were terminated in 1997, when they were disconnected from the water supply network.

Reservoirs - Brno, Zluty kopec (Yellow hill)

The diploma thesis deals with the creation of a design for a new building of the center of contemporary art and architecture and the use of existing reservoirs on Yellow Hill in Brno with the possibility of changing their original function. The work deals with the possibilities of processing the original terrain and the connection of the proposed new buildings to existing reservoirs. Architecturally, it explores the possibilities of designing new buildings contextually with the current architectural expression of the surrounding buildings and past architecture. The current situation is that a beautiful architectural and engineering monument, such as the reservoirs on Yellow Hill, is practically closed to the eyes of ordinary people. Only one of the three reservoirs is open to visit - the oldest brick reservoir built in 1872. The main goal of my work was therefore to make the reservoir area accessible to the public and to give people the opportunity to visit all three reservoirs. To this end, it was necessary to design entrance pavilions and safe entrances to the reservoirs and to fill the reservoirs with a new function, because the reservoirs were terminated in 1997, when they were disconnected from the water supply network