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New insights into the influence of pre-culture on robust solvent production of C. acetobutylicum
(2024)
Clostridia are known for their solvent production, especially the production of butanol. Concerning the projected depletion of fossil fuels, this is of great interest. The cultivation of clostridia is known to be challenging, and it is difficult to achieve reproducible results and robust processes. However, existing publications usually concentrate on the cultivation conditions of the main culture. In this paper, the influence of cryo-conservation and pre-culture on growth and solvent production in the resulting main cultivation are examined. A protocol was developed that leads to reproducible cultivations of Clostridium acetobutylicum. Detailed investigation of the cell conservation in cryo-cultures ensured reliable cell growth in the pre-culture. Moreover, a reason for the acid crash in the main culture was found, based on the cultivation conditions of the pre-culture. The critical parameter to avoid the acid crash and accomplish the shift to the solventogenesis of clostridia is the metabolic phase in which the cells of the pre-culture were at the time of inoculation of the main culture; this depends on the cultivation time of the pre-culture. Using cells from the exponential growth phase to inoculate the main culture leads to an acid crash. To achieve the solventogenic phase with butanol production, the inoculum should consist of older cells which are in the stationary growth phase. Considering these parameters, which affect the entire cultivation process, reproducible results and reliable solvent production are ensured.
Muscle function is compromised by gravitational unloading in space affecting overall musculoskeletal health. Astronauts perform daily exercise programmes to mitigate these effects but knowing which muscles to target would optimise effectiveness. Accurate inflight assessment to inform exercise programmes is critical due to lack of technologies suitable for spaceflight. Changes in mechanical properties indicate muscle health status and can be measured rapidly and non-invasively using novel technology. A hand-held MyotonPRO device enabled monitoring of muscle health for the first time in spaceflight (> 180 days). Greater/maintained stiffness indicated countermeasures were effective. Tissue stiffness was preserved in the majority of muscles (neck, shoulder, back, thigh) but Tibialis Anterior (foot lever muscle) stiffness decreased inflight vs. preflight (p < 0.0001; mean difference 149 N/m) in all 12 crewmembers. The calf muscles showed opposing effects, Gastrocnemius increasing in stiffness Soleus decreasing. Selective stiffness decrements indicate lack of preservation despite daily inflight countermeasures. This calls for more targeted exercises for lower leg muscles with vital roles as ankle joint stabilizers and in gait. Muscle stiffness is a digital biomarker for risk monitoring during future planetary explorations (Moon, Mars), for healthcare management in challenging environments or clinical disorders in people on Earth, to enable effective tailored exercise programmes.
Electronic cigarettes (e-cigarettes) have become popular worldwide with the market growing exponentially in some countries. The absence of product standards and safety regulations requires urgent development of analytical methodologies for the holistic control of the growing diversity of such products. An approach based on low-field nuclear magnetic resonance (LF-NMR) at 80 MHz is presented for the simultaneous determination of key parameters: carrier solvents (vegetable glycerine (VG), propylene glycol (PG) and water), total nicotine as well as free-base nicotine fraction. Moreover, qualitative and quantitative determination of fourteen weak organic acids deliberately added to enhance sensory characteristics of e-cigarettes was possible. In most cases these parameters can be rapidly and conveniently determined without using any sample manipulation such as dilution, extraction or derivatization steps. The method was applied for 37 authentic e-cigarettes samples. In particular, eight different organic acids with the content up to 56 mg/mL were detected. Due to its simplicity, the method can be used in routine regulatory control as well as to study release behaviour of nicotine and other e-cigarettes constituents in different products.
Frequency mixing magnetic detection (FMMD) is a sensitive and selective technique to detect magnetic nanoparticles (MNPs) serving as probes for binding biological targets. Its principle relies on the nonlinear magnetic relaxation dynamics of a particle ensemble interacting with a dual frequency external magnetic field. In order to increase its sensitivity, lower its limit of detection and overall improve its applicability in biosensing, matching combinations of external field parameters and internal particle properties are being sought to advance FMMD. In this study, we systematically probe the aforementioned interaction with coupled Néel–Brownian dynamic relaxation simulations to examine how key MNP properties as well as applied field parameters affect the frequency mixing signal generation. It is found that the core size of MNPs dominates their nonlinear magnetic response, with the strongest contributions from the largest particles. The drive field amplitude dominates the shape of the field-dependent response, whereas effective anisotropy and hydrodynamic size of the particles only weakly influence the signal generation in FMMD. For tailoring the MNP properties and parameters of the setup towards optimal FMMD signal generation, our findings suggest choosing large particles of core sizes dc > 25 nm nm with narrow size distributions (σ < 0.1) to minimize the required drive field amplitude. This allows potential improvements of FMMD as a stand-alone application, as well as advances in magnetic particle imaging, hyperthermia and magnetic immunoassays.
Wer A sagt, muss zumindest im Kaufrecht nicht immer B sagen: Es kommt nicht selten vor, dass sich in einem Kaufvertrag einerseits ein wirksamer Ausschluss der Gewährleistung des Verkäufers für Sachmängel findet, die Parteien aber andererseits gleichwohl eine Beschaffenheitsvereinbarung für bestimmte Eigenschaften vertraglich festlegen. In diesem Problemfeld führt eine aktuelle Entscheidung des BGH zu weiteren Klärungen für die Praxis (BGH, Urt. v. 10.4.2024 – VIII ZR 161/23, MDR 2024, 706). Der folgende Beitrag setzt sich mit den vielfältigen Aspekten der Entscheidung auseinander und erläutert, aus welchen Gründen der BGH dem Käufer einige goldene Brücken für einen Schadensersatzanspruch gebaut hat.
In der wasserbaulichen Forschung werden neben klassischen Messinstrumenten zunehmend kamerabasierte Verfahren genutzt. Diese erlauben neben der Bestimmung von Fließgeschwindigkeiten auch die Detektion der freien Wasseroberfläche oder zeitliche Vermessung von Kolken. Durch die hohen räumlichen und zeitlichen Auflösungen, welche neueste Kamerasensoren liefern, können neue Erkenntnisse in turbulenten, komplexen Strömungen gewonnen werden. Auch in der Praxis können diese Verfahren mit geringem Aufwand wichtige Daten liefern.
To gain insight on chemical sterilization processes, the influence of temperature (up to 70 °C), intense green light, and hydrogen peroxide (H₂O₂) concentration (up to 30% in aqueous solution) on microbial spore inactivation is evaluated by in-situ Raman spectroscopy with an optical trap. Bacillus atrophaeus is utilized as a model organism. Individual spores are isolated and their chemical makeup is monitored under dynamically changing conditions (temperature, light, and H₂O₂ concentration) to mimic industrially relevant process parameters for sterilization in the field of aseptic food processing. While isolated spores in water are highly stable, even at elevated temperatures of 70 °C, exposure to H₂O₂ leads to a loss of spore integrity characterized by the release of the key spore biomarker dipicolinic acid (DPA) in a concentration-dependent manner, which indicates damage to the inner membrane of the spore. Intensive light or heat, both of which accelerate the decomposition of H₂O₂ into reactive oxygen species (ROS), drastically shorten the spore lifetime, suggesting the formation of ROS as a rate-limiting step during sterilization. It is concluded that Raman spectroscopy can deliver mechanistic insight into the mode of action of H₂O₂-based sterilization and reveal the individual contributions of different sterilization methods acting in tandem.
Unmanned Aerial Vehicles (UAV) constantly gain in versatility. However, more reliable path planning algorithms are required until full autonomous UAV operation is possible. This work investigates the algorithm 3DVFH* and analyses its dependency on its cost function weights in 2400 environments. The analysis shows that the 3DVFH* can find a suitable path in every environment. However, a particular type of environment requires a specific choice of cost function weights. For minimal failure, probability interdependencies between the weights of the cost function have to be considered. This dependency reduces the number of control parameters and simplifies the usage of the 3DVFH*. Weights for costs associated with vertical evasion (pitch cost) and vicinity to obstacles (obstacle cost) have the highest influence on the failure probability of the local path planner. Environments with mainly very tall buildings (like large American city centres) require a preference for horizontal avoidance manoeuvres (achieved with high pitch cost weights). In contrast, environments with medium-to-low buildings (like European city centres) benefit from vertical avoidance manoeuvres (achieved with low pitch cost weights). The cost of the vicinity to obstacles also plays an essential role and must be chosen adequately for the environment. Choosing these two weights ideal is sufficient to reduce the failure probability below 10%.
Biomass from various types of organic waste was tested for possible use in hydrogen production. The composition consisted of lignified samples, green waste, and kitchen scraps such as fruit and vegetable peels and leftover food. For this purpose, the enzymatic pretreatment of organic waste with a combination of five different hydrolytic enzymes (cellulase, amylase, glucoamylase, pectinase and xylase) was investigated to determine its ability to produce hydrogen (H2) with the hydrolyzate produced here. In course, the anaerobic rod-shaped bacterium T. neapolitana was used for H2 production. First, the enzymes were investigated using different substrates in preliminary experiments. Subsequently, hydrolyses were carried out using different types of organic waste. In the hydrolysis carried out here for 48 h, an increase in glucose concentration of 481% was measured for waste loads containing starch, corresponding to a glucose concentration at the end of hydrolysis of 7.5 g·L−1. In the subsequent set fermentation in serum bottles, a H2 yield of 1.26 mmol H2 was obtained in the overhead space when Terrific Broth Medium with glucose and yeast extract (TBGY medium) was used. When hydrolyzed organic waste was used, even a H2 yield of 1.37 mmol could be achieved in the overhead space. In addition, a dedicated reactor system for the anaerobic fermentation of T. neapolitana to produce H2 was developed. The bioreactor developed here can ferment anaerobically with a very low loss of produced gas. Here, after 24 h, a hydrogen concentration of 83% could be measured in the overhead space.