Equilibrative Nucleoside Transporters Mediate the Import of Nicotinamide Riboside and Nicotinic Acid Riboside into Human Cells

Nicotinamide riboside (NR), a new form of vitamin B3, is an effective precursor of nicotinamide adenine dinucleotide (NAD+) in human and animal cells. The introduction of NR into the body effectively increases the level of intracellular NAD+ and thereby restores physiological functions that are weakened or lost in experimental models of aging and various pathologies. Despite the active use of NR in applied biomedicine, the mechanism of its transport into mammalian cells is currently not understood. In this study, we used overexpression of proteins in HEK293 cells, and metabolite detection by NMR, to show that extracellular NR can be imported into cells by members of the equilibrative nucleoside transporter (ENT) family ENT1, ENT2, and ENT4. After being imported into cells, NR is readily metabolized resulting in Nam generation. Moreover, the same ENT-dependent mechanism can be used to import the deamidated form of NR, nicotinic acid riboside (NAR). However, NAR uptake into HEK293 cells required the stimulation of its active utilization in the cytosol such as phosphorylation by NR kinase. On the other hand, we did not detect any NR uptake mediated by the concentrative nucleoside transporters (CNT) CNT1, CNT2, or CNT3, while overexpression of CNT3, but not CNT1 or CNT2, moderately stimulated NAR utilization by HEK293 cells.publishedVersio

Purine nucleoside phosphorylase controls nicotinamide riboside metabolism in mammalian cells

Nicotinamide riboside (NR) is an effective precursor of nicotinamide adenine dinucleotide (NAD) in human and animal cells. NR supplementation can increase the level of NAD in various tissues and thereby improve physiological functions that are weakened or lost in experimental models of aging or various human pathologies. However, there are also reports questioning the efficacy of NR supplementation. Indeed, the mechanisms of its utilization by cells are not fully understood. Herein, we investigated the role of purine nucleoside phosphorylase (PNP) in NR metabolism in mammalian cells. Using both PNP overexpression and genetic knockout, we show that after being imported into cells by members of the equilibrative nucleoside transporter family, NR is predominantly metabolized by PNP, resulting in nicotinamide (Nam) accumulation. Intracellular cleavage of NR to Nam is prevented by the potent PNP inhibitor Immucillin H in various types of mammalian cells. In turn, suppression of PNP activity potentiates NAD synthesis from NR. Combining pharmacological inhibition of PNP with NR supplementation in mice, we demonstrate that the cleavage of the riboside to Nam is strongly diminished, maintaining high levels of NR in blood, kidney, and liver. Moreover, we show that PNP inhibition stimulates Nam mononucleotide and NAD+ synthesis from NR in vivo, in particular, in the kidney. Thus, we establish PNP as a major regulator of NR metabolism in mammals and provide evidence that the health benefits of NR supplementation could be greatly enhanced by concomitant downregulation of PNP activity.publishedVersio

Nanohaloarchaea as beneficiaries of xylan degradation by haloarchaea

Climate change, desertification, salinisation of soils and the changing hydrology of the Earth are creating or modifying microbial habitats at all scales including the oceans, saline groundwaters and brine lakes. In environments that are saline or hypersaline, the biodegradation of recalcitrant plant and animal polysaccharides can be inhibited by salt-induced microbial stress and/or by limitation of the metabolic capabilities of halophilic microbes. We recently demonstrated that the chitinolytic haloarchaeon Halomicrobium can serve as the host for an ectosymbiont, nanohaloarchaeon ‘Candidatus Nanohalobium constans’. Here, we consider whether nanohaloarchaea can benefit from the haloarchaea-mediated degradation of xylan, a major hemicellulose component of wood. Using samples of natural evaporitic brines and anthropogenic solar salterns, we describe genome-inferred trophic relations in two extremely halophilic xylan-degrading three-member consortia. We succeeded in genome assembly and closure for all members of both xylan-degrading cultures and elucidated the respective food chains within these consortia. We provide evidence that ectosymbiontic nanohaloarchaea is an active ecophysiological component of extremely halophilic xylan-degrading communities (although by proxy) in hypersaline environments. In each consortium, nanohaloarchaea occur as ectosymbionts of Haloferax, which in turn act as scavenger of oligosaccharides produced by xylan-hydrolysing Halorhabdus. We further obtained and characterised the nanohaloarchaea–host associations using microscopy, multi-omics and cultivation approaches. The current study also doubled culturable nanohaloarchaeal symbionts and demonstrated that these enigmatic nano-sized archaea can be readily isolated in binary co-cultures using an appropriate enrichment strategy. We discuss the implications of xylan degradation by halophiles in biotechnology and for the United Nation's Sustainable Development Goals

Reduced TCA cycle rates at high hydrostatic pressure hinder hydrocarbon degradation and obligate oil degraders in natural, deep-sea microbial communities

Petroleum hydrocarbons reach the deep-sea following natural and anthropogenic factors. The process by which they enter deep-sea microbial food webs and impact the biogeochemical cycling of carbon and other elements is unclear. Hydrostatic pressure (HP) is a distinctive parameter of the deep sea, although rarely investigated. Whether HP alone affects the assembly and activity of oil-degrading communities remains to be resolved. Here we have demonstrated that hydrocarbon degradation in deep-sea microbial communities is lower at native HP (10 MPa, about 1000 m below sea surface level) than at ambient pressure. In long-term enrichments, increased HP selectively inhibited obligate hydrocarbon-degraders and downregulated the expression of beta-oxidation-related proteins (i.e., the main hydrocarbon-degradation pathway) resulting in low cell growth and CO2 production. Short-term experiments with HP-adapted synthetic communities confirmed this data, revealing a HP-dependent accumulation of citrate and dihydroxyacetone. Citrate accumulation suggests rates of aerobic oxidation of fatty acids in the TCA cycle were reduced. Dihydroxyacetone is connected to citrate through glycerol metabolism and glycolysis, both upregulated with increased HP. High degradation rates by obligate hydrocarbon-degraders may thus be unfavourable at increased HP, explaining their selective suppression. Through lab-scale cultivation, the present study is the first to highlight a link between impaired cell metabolism and microbial community assembly in hydrocarbon degradation at high HP. Overall, this data indicate that hydrocarbons fate differs substantially in surface waters as compared to deep-sea environments, with in situ low temperature and limited nutrients availability expected to further prolong hydrocarbons persistence at deep sea

Enzymes for consumer products to achieve climate neutrality

29 pags., 4 figs., 3 tabs., 1 graf.Accumulated greenhouse gas emissions are expected to increase from 36.2 Giga-tons (Gt) to 60 Gt over the next three decades. The global surface temperature has increased by¿+¿1.09¿°C since 2001, and might increase by¿+¿2.2¿°C in 2100, +3.6¿°C in 2200 and +4.6¿°C in 2500. These emissions and temperature rises cannot be reduced in their entirety, but they can be lowered by using enzymes. Enzymes are proteins that catalyze biochemical reactions that make life possible since 3.8 billion years ago. Scientists have been able to "domesticate" them in such a way that enzymes, and their engineered variants, are now key players of the circular economy. With a world production of 117 Kilo-tons and a trade of 14.5 Billion-dollars, they have the potential to annually decrease CO2 emissions by 1 to 2.5 Billion-tons (Bt), the carbon demand to synthesise chemicals by 200 Million tons (Mt), the amount of chemicals by 90¿Mt, and the economic losses derived from global warming by 0.5%, while promoting biodiversity and our planet¿s health. Our success to increase these benefits will depend on better integration of enzymatic solutions in different sectors.This study was conducted under the auspices of the FuturEnzyme Project funded by the European Union’s Horizon 2020 Research and Innovation Programme under Grant Agreement No. 101000327. MF also acknowledges Grants PID2020-112758RB-I00, PDC2021-121534-I00, and TED2021-130544B-I00 from the MCIN/AEI/10.13039/501100011033 and the European Union (“NextGenerationEU/PRTR”)

INTERNET TECHNOLOGIES FOR TRAFFIC AND PEDESTRIAN DATA COLLECTION

The article shows the technology of obtaining initial data on the state and functioning of the city transport system. The technology makes it possible to obtain data on the intensity of any movement: intensity of traffic, pedestrians and passengers. The main idea is to form and monitor the implementation of the monitoring program, during which it is possible to elaborate in detail the entire procedure for collecting information, including the dislocation of places for collecting the intensity of traffic, passengers and pedestrians, including the time of their collection. At the stage of direct data collection software provides clear unambiguous instructions for each accountant involved in the survey, about what actions at the given time it needs to take. The large volume of preparatory and organizational activities given out by the hardware and software allows using the technology to significantly simplify the issues of monitoring the technology of information collected, as well as the culling of incorrectly collected information, by monitoring the location of the accountant, controlling the start and end of data collection. In particular, the technology has proven itself to calculate the intensity of traffic flows when collecting such information from urban intersections. The presented technology allows carrying out large-scale surveys of transport, passenger and pedestrian flows in cities with high efficiency at the stage of construction of forecast and optimization transport models of cities. The software interface of the mobile and server part of the presented technology allows achieving automatic interface of the collected data with the necessary format for their further use as calibration information of forecast transport models implemented in modern software products such as PTV Vision VISUM

Analytical Study of QoS-Oriented Multicast in Wireless Networks

<p/> <p>Multicast is a very popular bandwidth-conserving technology exploited in many multimedia applications. However, existing standards of high rate wireless networks provide no error recovery mechanism (ARQ) for multicast traffic. ARQ absence in wireless networks unreliable by their nature leads to frequent packet losses, which is inappropriate for most of multimedia applications. In this paper, we study new reliable multicast mechanism proposed recently to support multimedia QoS (packet loss ratio, latency, and throughput) with various wireless technologies. This mechanism is based on the concept of multiple ACK-leaders, that is, multicast recipients responsible for acknowledging data packets. We develop analytical models of the mechanism with various leader selection schemes and use the models to study the schemes efficiency and to optimize them. Numerical results show that the novel multicast mechanism with multiple ACK-leaders can be easily tuned to meet specific QoS requirements of multimedia or any other multicast applications.</p

Symmetrization and Amplification of Germicidal Radiation Flux Produced by a Mercury Amalgam UV Lamp in Cylindrical Cavity with Diffusely Reflective Walls

The study focused on increasing the efficiency of germicidal UV radiation by using highly diffuse reflective materials such as PTFE in irradiated cavities of UV air purifiers. In a conventional cylindrically symmetric cavity with a linear amalgam mercury lamp as UV-radiation source on the axis UV-radiation, flux directed from the lamp to the walls dropped from the axis to the periphery. To increase the UV irradiation, the walls are often made mirror-reflective, but the radiation flux distribution remained radially symmetric with a maximum on the source emitting surface in this case as well. When most of the emitted light is returned to the source after one reflection, the conditions of its operation are disturbed. If the walls are made of highly diffuse reflective materials, the radiation flux density inside the cavity increases on average, and its distribution becomes uniform and highly symmetric. Thus, the effect of amplification of the radiation flux due to the highly diffuse reflectivity of the walls increases with radius and reaches a maximum at the wall. Experiments were performed to demonstrate increasing amplification of germicidal UV radiation flux with a diffuse reflection coefficient in cylindrical cavities with walls of PTFE and ePTFE. The irradiation of the cavity wall was observed to increase up to 20 times at the resonant mercury line of 253.7 nm and up to 40 times at some non-resonant lines of the visible range due to highly diffuse reflectivity of the cavity walls. The flux amplification effect was limited by the diffuse reflectivity value of the walls and absorption coefficient of the radiation emitting surface. A formula for calculating the radiation flux amplification factor in a diffusely reflecting cylindrically symmetric cavity was derived for the case of Lambertian source and reflector, including wall reflectivity and source surface absorption coefficients. The effects of heating and cooling of the mercury lamp amalgam directly affected the amplification, and symmetrization of germicidal irradiation was observed and is discussed in the paper. Numerical calculations were performed by the ray tracing method. The calculated model was verified by comparing the numerical results with those of both the approximate theoretical consideration and experiments. The promising use of diffusely reflecting cylindrical cavities for UV air purifiers is discussed. Designs of air inlet and outlet ports that allow effective locking of germicidal radiation inside the UV air purifiers were considered. The results of this work may be of interest for further developments in the UV disinfection technique

Combined Surface Heating by Laser Beam and Subsonic Nitrogen Plasma Jet

The paper describes new combined heating capability of the IPMech RAS inductively coupled plasma facility VGU-4. A 200 W ytterbium laser was added to the facility as a source of radiative heating. The cylindrical specimen made of the Buran orbital vehicle’s heat-shielding tile material with a black low catalytic coating was exposed to subsonic pure nitrogen plasma jet and laser radiation. The specimen surface temperature reached 1325 ∘C during combined radiative and convective heating. The maximum heat flux obtained in the combined mode for a laser incident power of 47 W and a VGU-4 HF-generator anode power of 22 kW was 32.1 W/cm2. The convective heat flux from the nitrogen plasma jet at the same anode power was 12.6 W/cm2. Adding a laser to an existing inductively coupled plasma facility gives the future opportunity to better simulate entry into the atmospheres of Mars, Venus, the outer planets and their moons