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Background
Minor changes in protein structure induced by small organic and inorganic molecules can result in significant metabolic effects. The effects can be even more profound if the molecular players are chemically active and present in the cell in considerable amounts. The aim of our study was to investigate effects of a nitric oxide donor (spermine NONOate), ATP and sodium/potassium environment on the dynamics of thermal unfolding of human hemoglobin (Hb). The effect of these molecules was examined by means of circular dichroism spectrometry (CD) in the temperature range between 25°C and 70°C. The alpha-helical content of buffered hemoglobin samples (0.1 mg/ml) was estimated via ellipticity change measurements at a heating rate of 1°C/min.
Results
Major results were:
1) spermine NONOate persistently decreased the hemoglobin unfolding temperature T u irrespectively of the Na + /K + environment,
2) ATP instead increased the unfolding temperature by 3°C in both sodium-based and potassium-based buffers and
3) mutual effects of ATP and NO were strongly influenced by particular buffer ionic compositions. Moreover, the presence of potassium facilitated a partial unfolding of alpha-helical structures even at room temperature.
Conclusion
The obtained data might shed more light on molecular mechanisms and biophysics involved in the regulation of protein activity by small solutes in the cell.
One of the priority trends of carbon nanotechnology is creation of nanocomposite systems. Such carbon nanostructured composites were produced using - raw materials based on the products of agricultural waste, such as grape stones, apricot stones, rice husk. These products have a - wide spectrum of application and can be obtained in large quantities. The Institute of Combustion Problems has carried out the work on synthesis of the nanostructured carbon sorbents for multiple applications including the field of biomedicine. The article presents the data on the synthesis and physico-chemical properties of carbonaceous sorbents using physicochemical methods of investigation: separation and purification of biomolecules; isolation of phytohormone - fusicoccin; adsorbent INGO-1 in the form of an adsorption column for blood detoxification, oral (entero) sorbent - INGO-2; the study of efferent and probiotic properties and sorption activity in regard to the lipopolysaccharide (LPS), new biocomposites - based on carbonized rice husk (CRH) and cellular microorganisms; the use of CRH in wound treatment. A new material for blood detoxication (INGO-1) has been obtained. Adsorption of p-cresyl sulfate and indoxyl sulfate has shown that active carbon adsorbent can remove clinically significant level of p-cresyl sulfate and indoxyl sulfate from human plasma. Enterosorbent INGO-2 possesses high adsorption activity in relation to Gram-negative bacteria and their endotoxins. INGO-2 slows down the growth of conditionally pathogenic microorganisms, without having a negative effect on bifido and lactobacteria. The use of enterosorbent INGO-2 for sorption therapy may provide a solution to a complex problem - detoxication of the digestive tract and normalization of the intestinal micro ecology. The immobilized probiotic called "Riso-lact" was registered at the Ministry of Health of the Republic of Kazakhstan as a biologically active food additive. The developed technology is patented and provides production of the medicine in the form of freeze-dried biomass immobilized in vials.
Autonomous robotic systems for penetrating thick ice shells with simultaneous collecting of scientific data are very promising devices in both terrestrial (glacier, climate research) and extra-terrestrial applications. Technical challenges in development of such systems are numerous and include 3D-navigation, an appropriate energy source, motion control, etc. Not less important is the problem of forward contamination of the pristine glacial environments with microorganisms and biomolecules from the surface of the probe. This study was devoted to establishing a laboratory model for microbial contamination of a newly constructed ice-melting probe called IceMole and to analyse the viability and amount of the contaminating microorganisms as a function of distance. The used bacterial strains were Bacillus subtilis (ATCC 6051) and Escherichia coli (ATCC 11775). The main objective was development of an efficient and reliable in-situ decontamination method of the melting probe. Therefore, several chemical substances were tested in respect of their efficacy to eliminate bacteria on the surface of the melting probe at low temperature (0 - 5 °C) and at continuous dilution by melted water. Our study has shown that at least 99.9% decontamination of the IceMole can be successfully achieved by the injection of 30% (v/v) hydrogen peroxide and 3% (v/v) sodium hypochlorite into the drilling site. We were able to reproduce this result in both time-dependent and depth-dependent experiments. The sufficient amount of 30% (v/v) H₂O₂ or 3% (v/v) NaClO has been found to be approximately 18 L per cm² of the probe’s surface.
Suspension depletion approach for exemption of infected Solanum jasminoides cells from pospiviroids
(2018)
Despite numerous studies, viroid elimination from infected plants remains a very challenging task. This study introduces for the first time a novel ‘suspension depletion’ approach for exemption of Solanum jasminoides plants from viroids. The proposed method implies initial establishment of suspension cultures of the infected plant cells. The suspended cells were then physically treated (mild thermotherapy, 33 °C), which presumably delayed the replication of the viroid. The viroid concentration in the treated biomass was monitored weekly using pospiviroid-specific PCR. After 10–12 weeks of continuous treatment, a sufficient decrease in viroid concentration was observed such that the infection became undetectable by PCR. The treated single cells then gave rise to microcolonies on a solid culture medium and the obtained viroid-negative clones were further promoted to regenerate into viroid-free plants. Three years of accumulated experimental data suggests feasibility, broad applicability, and good efficacy of the proposed approach.
We present the novel concept of a combined drilling and melting probe for subsurface ice research. This probe, named “IceMole”, is currently developed, built, and tested at the FH Aachen University of Applied Sciences’ Astronautical Laboratory. Here, we describe its first prototype design and report the results of its field tests on the Swiss Morteratsch glacier. Although the IceMole design is currently adapted to terrestrial glaciers and ice shields, it may later be modified for the subsurface in-situ investigation of extraterrestrial ice, e.g., on Mars, Europa, and Enceladus. If life exists on those bodies, it may be present in the ice (as life can also be found in the deep ice of Earth).
There is significant interest in sampling subglacial environments for geobiological studies, but they are difficult to access. Existing ice-drilling technologies make it cumbersome to maintain microbiologically clean access for sample acquisition and environmental stewardship of potentially fragile subglacial aquatic ecosystems. The IceMole is a maneuverable subsurface ice probe for clean in situ analysis and sampling of glacial ice and subglacial materials. The design is based on the novel concept of combining melting and mechanical propulsion. It can change melting direction by differential heating of the melting head and optional side-wall heaters. The first two prototypes were successfully tested between 2010 and 2012 on glaciers in Switzerland and Iceland. They demonstrated downward, horizontal and upward melting, as well as curve driving and dirt layer penetration. A more advanced probe is currently under development as part of the Enceladus Explorer (EnEx) project. It offers systems for obstacle avoidance, target detection, and navigation in ice. For the EnEx-IceMole, we will pay particular attention to clean protocols for the sampling of subglacial materials for biogeochemical analysis. We plan to use this probe for clean access into a unique subglacial aquatic environment at Blood Falls, Antarctica, with return of a subglacial brine sample.
The Saturnian moon Enceladus with its extensive water bodies underneath a thick ice sheet cover is a potential candidate for extraterrestrial life. Direct exploration of such extraterrestrial aquatic ecosystems requires advanced access and sampling technologies with a high level of autonomy. A new technological approach has been developed as part of the collaborative research project Enceladus Explorer (EnEx). The concept is based upon a minimally invasive melting probe called the IceMole. The force-regulated, heater-controlled IceMole is able to travel along a curved trajectory as well as upwards. Hence, it allows maneuvers which may be necessary for obstacle avoidance or target selection. Maneuverability, however, necessitates a sophisticated on-board navigation system capable of autonomous operations. The development of such a navigational system has been the focal part of the EnEx project. The original IceMole has been further developed to include relative positioning based on in-ice attitude determination, acoustic positioning, ultrasonic obstacle and target detection integrated through a high-level sensor fusion. This paper describes the EnEx technology and discusses implications for an actual extraterrestrial mission concept.
The ”IceMole“ is a novel maneuverable subsurface ice probe for clean in-situ analysis and sampling of subsurface ice and subglacial water/brine. It is developed and build at FH Aachen University of Applied Sciences’ Astronautical Laboratory. A first prototype was successfully tested on the Swiss Morteratsch glacier in 2010. Clean sampling is achieved with a hollow ice screw (as it is used in mountaineering) at the tip of the probe. Maneuverability is achieved with a differentially heated melting head. Funded by the German Space Agency (DLR), a consortium led by FH Aachen currently develops a much more advanced IceMole probe, which includes a sophisticated system for obstacle avoidance, target detection, and navigation in the ice. We intend to use this probe for taking clean samples of subglacial brine at the Blood Falls (McMurdo Dry Valleys, East Antarctica) for chemical and microbiological analysis. In our conference contribution, we 1) describe the IceMole design, 2) report the results of the field tests of the first prototype on the Morteratsch glacier, 3) discuss the probe’s potential for the clean in-situ analysis and sampling of subsurface ice and subglacial liquids, and 4) outline the way ahead in the development of this technology.
Subglacial environments on Earth offer important analogs to Ocean World targets in our solar system. These unique microbial ecosystems remain understudied due to the challenges of access through thick glacial ice (tens to hundreds of meters). Additionally, sub-ice collections must be conducted in a clean manner to ensure sample integrity for downstream microbiological and geochemical analyses. We describe the field-based cleaning of a melt probe that was used to collect brine samples from within a glacier conduit at Blood Falls, Antarctica, for geomicrobiological studies. We used a thermoelectric melting probe called the IceMole that was designed to be minimally invasive in that the logistical requirements in support of drilling operations were small and the probe could be cleaned, even in a remote field setting, so as to minimize potential contamination. In our study, the exterior bioburden on the IceMole was reduced to levels measured in most clean rooms, and below that of the ice surrounding our sampling target. Potential microbial contaminants were identified during the cleaning process; however, very few were detected in the final englacial sample collected with the IceMole and were present in extremely low abundances (∼0.063% of 16S rRNA gene amplicon sequences). This cleaning protocol can help minimize contamination when working in remote field locations, support microbiological sampling of terrestrial subglacial environments using melting probes, and help inform planetary protection challenges for Ocean World analog mission concepts.