Search

Cell culture on orbit is complicated by numerous operational constraints, including g-loads on the ascent, vibrations, transit time to International Space Station, and delays in experiment initiation. Cryopreserving cells before launch would negate these factors. However, defrosting these cells is problematic, since the traditional method of employing a water bath is not possible. We here describe a unique apparatus designed to accomplish this in a microgravitational environment. This apparatus resulted in rapid defrost of cryopreserved cell cultures and allowed successful tissue culture operations on the station for periods of up to 21 days. [ABSTRACT FROM AUTHOR]

Discriminatory analysis of aroma compounds on basis of statistical and enological paramaters, for sensory evaluation of beverages made from grapes, is provided.

The aroma of Oloroso sherry wines obtained by industrial oxidative aging was studied by gas chromatography-olfactometry analysis. The aromagram exhibited five different odorant zones (OZ) separated by no-odor intervals of at least 6 min, in which no odor was detected. Based on the perceived aroma frequency the most distinctive series for OZ1was fruity. The OZ2and OZ3were mainly represented by chemical and fatty series, respectively, and the spicy and empyreumatic series were the most representative for the OZ4and OZ5, although exhibiting few notes. Except for the vegetable, floral, and empyreumatic series, the remainder increased the number of odorant notes perceived during aging. [ABSTRACT FROM PUBLISHER]

The aroma of Fino sherry wines produced by industrial biological aging for 0, 1.5, 2.5, 4.5, and 6 years in the Montilla-Moriles region (southern Spain) was studied by gas chromatography-olfactometry. The aroma sensations detected by 3 trained sniffers were classified according to their odor descriptors into 8 odorant series (fruity, empyreumatic, chemical, fatty, balsamic, vegetable, floral, and spicy), describing the aroma profile of the studied wines. The results showed 47 detected odors in the unaged wines, 50 in the 1.5-years-old wines and 59, 61 and 69 in the wines aged 2.5, 4.5, and 6 years, respectively. According to the frequency of the perceived aromas, the fruity and empyreumatic series were the most characteristic odorant series. By exception of chemical, floral and balsamic series without changes during aging of the wines, the remainder series increased their participation during the aging, mainly the fruity, empyreumatic, and fatty series. [ABSTRACT FROM AUTHOR]

We report the case of a 64 year-old woman with motor and cognitive deterioration that progressed rapidly during eight months. She was unsuccessfully treated with quinacrine, and died in a terminal status, by septic shock secondary to bronchopneumonia by broncho-aspiration. The brain was donated for research and the histopathological analysis showed spongiform changes, astrogliosis and prion protein (PrPRes) deposits, confirmed by Western blot (WB). These features are considered characteristic of prion diseases, which are uncommon in Colombia. We highlight that its diagnosis was made for the first time in this country by the simultaneous use of immunohistochemistry and Western blot.

Faecal samples were obtained from 190 children, aged 0 to 5 years, admitted to a public hospital in Belém, Pará, Brazil. These patients were placed in a pediatric ward with 40 beds distributed in six rooms. Case were classified into three groups: (a) nosocomial: children who developed gastroenteritis 72 hr or later after admission; (b) community-acquired: patients admitted either with diarrhoea or who had diarrhoea within 72 hr following admission; (c) non-diarrhoeic: those children who had no diarrhoea three days before and three days after collection of formed faecal sample. Specimens were routinely processed for the presence of rotaviruses, bacteria and parasites. Rotaviruses were detected through enzyme-linked immunosorbent assay (ELISA) and subsequently serotyped/electrophoretyped. Rotaviruses were the most prevalent enteropathogens among nosocomial cases, accounting for 39 % (9/23) of diarrhoeal episodes; on the other hand, rotaviruses ocurred in 8.3 % (11/133) and 9 % (3/34) of community-acquired and non-diarrhoeic categories, respectively. Mixed infections involving rotavirus and Giardia intestinalis and rotavirus plus G. intestinalis and Entamoeba histolytica were detected in frequencies of 8.6 and 4.3 %, respectively, in the nosocomial group. The absence of bacterial pathogens in this category, and the unusual low prevalence of these agents in the other two groups may reflect the early and routine administration of antibiotics following admission to this hospital. Rotavirus serotype 2 prevailed over the other types, accounting for 77.8 % of isolates from nosocomial diarrhoeal episodes. In addition, at least five different genomic profiles could be observed, of which one displayed an unusual five-segment first RNA cluster. Dehydration was recorded in all cases of hospital-acquired, rotavirus-associated diarrhoea, whereas in only 57 % of nosocomial cases of other aetiology. It was also noted that nosocomial, rotavirus-associated diarrhoeal episodes occur earlier (7 days), following admission, if compared with those hospital-acquired cases of other aetiology (14 days).

The recent acceleration of commercial, private and multi-national spaceflight has created an unprecedented level of activity in low Earth orbit, concomitant with the largest-ever number of crewed missions entering space and preparations for exploration-class (lasting longer than one year) missions. Such rapid advancement into space from many new companies, countries and space-related entities has enabled a 'second space age'. This era is also poised to leverage, for the first time, modern tools and methods of molecular biology and precision medicine, thus enabling precision aerospace medicine for the crews. The applications of these biomedical technologies and algorithms are diverse, and encompass multi-omic, single-cell and spatial biology tools to investigate human and microbial responses to spaceflight. Additionally, they extend to the development of new imaging techniques, real-time cognitive assessments, physiological monitoring and personalized risk profiles tailored for astronauts. Furthermore, these technologies enable advancements in pharmacogenomics, as well as the identification of novel spaceflight biomarkers and the development of corresponding countermeasures. In this Perspective, we highlight some of the recent biomedical research from the National Aeronautics and Space Administration, Japan Aerospace Exploration Agency, European Space Agency and other space agencies, and detail the entrance of the commercial spaceflight sector (including SpaceX, Blue Origin, Axiom and Sierra Space) into aerospace medicine and space biology, the first aerospace medicine biobank, and various upcoming missions that will utilize these tools to ensure a permanent human presence beyond low Earth orbit, venturing out to other planets and moons., (© 2024. Springer Nature Limited.)

As the International Space Station comes to the end of a transformative era of in-space research, NASA's Commercial Low Earth Orbit (LEO) Destinations (CLD) Program aims to catalyze a new generation of platforms with co-investment from the private sector, preventing a potential gap in research performed in LEO, while building a robust LEO economy. In this paper, we provide insight into the CLD Program focusing on Orbital Reef, describing its operational and technical characteristics as well as new opportunities it may enable. Achieving about a third of the pressurized volume of the ISS with the launch of a single pressurized module and growing to support hundreds of Middeck Locker Equivalents (MLE) in passive and active payloads internally and externally, Orbital Reef will enable government, academic, and commercial institutions to continue and expand upon research and development (R&D) efforts currently performed on ISS. Additionally, it will enable nascent markets to establish their operations in space, by initiating new lines of research and technology development and the implementation of new ventures and visions. Using Blue Origin's New Glenn heavy launch system, Sierra Space's cargo and crew Dream Chaser® vehicles, and Boeing's Starliner crew vehicle, and expertise from Amazon/Amazon Supply Chain, Arizona State University, Genesis Engineering, and Redwire, Orbital Reef is being designed to address ISS-era transportation logistics challenges. Finally, this manuscript describes some of the expected challenges from the ISS-to-CLD transition, and provides guidance on how researchers in academia and industry can shape the future of commercial destinations and work performed in LEO., (© 2024. The Author(s).)

Bacterial growth and behavior have been studied in microgravity in the past, but little focus has been directed to cell size despite its impact on a myriad of processes, including biofilm formation, which is impactful regarding crew health. To interrogate this characteristic, supernatant aliquots of P. aeruginosa cultured on different materials and media on board the International Space Station (ISS) as part of the Space Biofilms Project were analyzed. For that experiment, P. aeruginosa was grown in microgravity-with matching Earth controls-in modified artificial urine medium (mAUMg-high Pi) or LB Lennox supplemented with KNO 3 , and its formation of biofilms on six different materials was assessed. After one, two, and three days of incubation, the ISS crew terminated subsets of the experiment by fixation in paraformaldehyde, and aliquots of the supernatant were used for the planktonic cell size study presented here. The measurements were obtained post-flight through the use of phase contrast microscopy under oil immersion, a Moticam 10+ digital camera, and the FIJI image analysis program. Statistical comparisons were conducted to identify which treatments caused significant differences in cell dimensions using the Kruskal-Wallis and Dunn tests. There were statistically significant differences as a function of material present in the culture in both LBK and mAUMg-high Pi. Along with this, the data were also grouped by gravitational condition, media, and days of incubation. Comparison of planktonic cells cultured in microgravity showed reduced cell length (from 4% to 10% depending on the material) and diameter (from 1% to 10% depending on the material) with respect to their matching Earth controls, with the caveat that the cultures may have been at different points in their growth curve at a given time. In conclusion, smaller cells were observed on the cultures grown in microgravity, and cell size changed as a function of incubation time and the material upon which the culture grew. We describe these changes here and possible implications for human space travel in terms of crew health and potential applications.

Microorganisms' natural ability to live as organized multicellular communities - also known as biofilms - provides them with unique survival advantages. For instance, bacterial biofilms are protected against environmental stresses thanks to their extracellular matrix, which could contribute to persistent infections after treatment with antibiotics. Bacterial biofilms are also capable of strongly attaching to surfaces, where their metabolic by-products could lead to surface material degradation. Furthermore, microgravity can alter biofilm behavior in unexpected ways, making the presence of biofilms in space a risk for both astronauts and spaceflight hardware. Despite the efforts to eliminate microorganism contamination from spacecraft surfaces, it is impossible to prevent human-associated bacteria from eventually establishing biofilm surface colonization. Nevertheless, by understanding the changes that bacterial biofilms undergo in microgravity, it is possible to identify key differences and pathways that could be targeted to significantly reduce biofilm formation. The bacterial component of Space Biofilms project, performed on the International Space Station in early 2020, contributes to such understanding by characterizing the morphology and gene expression of bacterial biofilms formed in microgravity with respect to ground controls. Pseudomonas aeruginosa was used as model organism due to its relevance in biofilm studies and its ability to cause urinary tract infections as an opportunistic pathogen. Biofilm formation was characterized at one, two, and three days of incubation (37 °C) over six different materials. Materials reported in this manuscript include catheter grade silicone, selected due to its medical relevance in hospital acquired infections, catheter grade silicone with ultrashort pulsed direct laser interference patterning, included to test microtopographies as a potential biofilm control strategy, and cellulose membrane to replicate the column and canopy structure previously reported from a microgravity study. We here present an overview of the biofilm morphology, including 3D images of the biofilms to represent the distinctive morphology observed in each material tested, and some of the key differences in biofilm thickness, mass, and surface area coverage. We also present the impact of the surface microtopography in biofilm formation across materials, incubation time, and gravitational conditions. The Space Biofilms project (bacterial side) is supported by the National Aeronautics and Space Administration under Grant No. 80NSSC17K0036 and 80NSSC21K1950., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

In microgravity, bacteria undergo intriguing physiological adaptations. There have been few attempts to assess global bacterial physiological responses to microgravity, with most studies only focusing on a handful of individual systems. This study assessed the fitness of each gene in the genome of the aromatic compound-degrading Alphaproteobacterium Novosphingobium aromaticavorans during growth in spaceflight. This was accomplished using Comparative TnSeq, which involves culturing the same saturating transposon mutagenized library under two different conditions. To assess gene fitness, a novel comparative TnSeq analytical tool was developed, named TnDivA, that is particularly useful in leveraging biological replicates. In this approach, transposon diversity is represented numerically using a modified Shannon diversity index, which was then converted into effective transposon density. This transformation accounts for variability in read distribution between samples, such as cases where reads were dominated by only a few transposon inserts. Effective density values were analyzed using multiple statistical methods, including log 2 -fold change, least-squares regression analysis, and Welch's t-test. The results obtained across applied statistical methods show a difference in the number of significant genes identified. However, the functional categories of genes important to growth in microgravity showed similar patterns. Lipid metabolism and transport, energy production, transcription, translation, and secondary metabolite biosynthesis and transport were shown to have high fitness during spaceflight. This suggests that core metabolic processes, including lipid and secondary metabolism, play an important role adapting to stress and promoting growth in microgravity., (© 2023. The Author(s).)

The undesirable, yet inevitable, presence of bacterial biofilms in spacecraft poses a risk to the proper functioning of systems and to astronauts' health. To mitigate the risks that arise from them, it is important to understand biofilms' behavior in microgravity. As part of the Space Biofilms project, biofilms of Pseudomonas aeruginosa were grown in spaceflight over material surfaces. Stainless Steel 316 (SS316) and passivated SS316 were tested for their relevance as spaceflight hardware components, while a lubricant impregnated surface (LIS) was tested as potential biofilm control strategy. The morphology and gene expression of biofilms were characterized. Biofilms in microgravity are less robust than on Earth. LIS strongly inhibits biofilm formation compared to SS. Furthermore, this effect is even greater in spaceflight than on Earth, making LIS a promising option for spacecraft use. Transcriptomic profiles for the different conditions are presented, and potential mechanisms of biofilm reduction on LIS are discussed., (© 2023. Springer Nature Limited.)

Fungi biofilms have been found growing on spacecraft surfaces such as windows, piping, cables, etc. The contamination of these surfaces with fungi, although undesirable, is highly difficult to avoid. While several biofilm forming species, including Penicillium rubens , have been identified in spacecraft, the effect of microgravity on fungal biofilm formation is unknown. This study sent seven material surfaces (Stainless Steel 316, Aluminum Alloy, Titanium Alloy, Carbon Fiber, Quartz, Silicone, and Nanograss) inoculated with spores of P. rubens to the International Space Station and allowed biofilms to form for 10, 15, and 20 days to understand the effects of microgravity on biofilm morphology and growth. In general, microgravity did not induce changes in the shape of biofilms, nor did it affect growth in terms of biomass, thickness, and surface area coverage. However, microgravity increased or decreased biofilm formation in some cases, and this was incubation-time- and material-dependent. Nanograss was the material with significantly less biofilm formation, both in microgravity and on Earth, and it could potentially be interfering with hyphal adhesion and/or spore germination. Additionally, a decrease in biofilm formation at 20 days, potentially due to nutrient depletion, was seen in some space and Earth samples and was material-dependent.

Finding sustainable approaches to achieve independence from terrestrial resources is of pivotal importance for the future of space exploration. This is relevant not only to establish viable space exploration beyond low Earth-orbit, but also for ethical considerations associated with the generation of space waste and the preservation of extra-terrestrial environments. Here we propose and highlight a series of microbial biotechnologies uniquely suited to establish sustainable processes for in situ resource utilization and loop-closure. Microbial biotechnologies research and development for space sustainability will be translatable to Earth applications, tackling terrestrial environmental issues, thereby supporting the United Nations Sustainable Development Goals., (© 2023. The Author(s).)

Concentrated agave sap is a product with in vivo proven hypocholesterolemic and hypoglycemic activities, as well as in vitro anticancer potential. In the present work, a factorial design was used to determine the suitable drying conditions of concentrated agave by studying the effect of inlet temperature (150 °C, 180 °C and 210 °C) and the type of carrier agent (maltodextrin, hydroxypropyl methylcellulose, guar gum and xanthan gum). The response variables for each treatment were the product recovery and microencapsulated saponins. Further characterization of concentrated agave powders was performed: solubility in water, hygroscopicity, moisture content, tap density, bulk density, Carr's index followability and morphology by scanning electron microscopy analysis. The hydroxypropyl methylcellulose proved to improve physicochemical properties and enhance product yield, using 210 °C inlet temperature and a mix of carrier agents of maltodextrin/hydroxypropyl methylcellulose/xanthan gum at 50/48.5/1.5 (w/w/w) proportion exhibited the highest saponin recovery of 53.81%. Moreover, different carrier agents in powders revealed two shapes, regular spherical shape with smooth surface and collapsed shapes. The use of polymers excipients helped to decrease the stickiness of the desired product and enhanced the powder stability and microencapsulation of the steroidal saponins.

Bacterial behavior has been studied under microgravity conditions, but very little is known about it under lunar and Martian gravitational regimes. An Earth-based approach was designed and implemented using inclined clinostats and an in-house-developed code to determine the optimal clinorotation angular speed for bacterial liquid cultures of 5 RPM. With this setup, growth dynamics, phenotypic changes, and sensitivity to antibiotics (minimum inhibitory concentration (MIC) of two different classes of antibiotics) for three Escherichia coli strains (including uropathogenic) were examined under simulated micro-, lunar, and Martian gravities. The results included increased growth under simulated micro- and lunar gravities for some strains, and higher concentrations of antibiotics needed under simulated lunar gravity with respect to simulated micro- and Martian gravities. Clinostat-produced results can be considered suggestive but not determinative of what might be expected in altered gravity, as there is still a need to systematically verify these simulation devices' ability to accurately replicate phenomena observed in space. Nevertheless, this approach serves as a baseline to start interrogating key cellular and molecular aspects relevant to microbial processes on the lunar and Martian surfaces.

As we aim to expand human presence in space, we need to find viable approaches to achieve independence from terrestrial resources. Space biomining of the Moon, Mars and asteroids has been indicated as one of the promising approaches to achieve in-situ resource utilization by the main space agencies. Structural and expensive metals, essential mineral nutrients, water, oxygen and volatiles could be potentially extracted from extraterrestrial regolith and rocks using microbial-based biotechnologies. The use of bioleaching microorganisms could also be applied to space bioremediation, recycling of waste and to reinforce regenerative life support systems. However, the science around space biomining is still young. Relevant differences between terrestrial and extraterrestrial conditions exist, including the rock types and ores available for mining, and a direct application of established terrestrial biomining techniques may not be a possibility. It is, therefore, necessary to invest in terrestrial and space-based research of specific methods for space applications to learn the effects of space conditions on biomining and bioremediation, expand our knowledge on organotrophic and community-based bioleaching mechanisms, as well as on anaerobic biomining, and investigate the use of synthetic biology to overcome limitations posed by the space environments., (© 2021. The Author(s).)

The biomining microbes which extract metals from ores that have been applied in mining processes worldwide hold potential for harnessing space resources. Their cell growth and ability to extract metals from extraterrestrial minerals under microgravity environments, however, remains largely unknown. The present study used the model biomining bacterium Acidithiobacillus ferrooxidans to extract metals from lunar and Martian regolith simulants cultivated in a rotating clinostat with matched controls grown under the influence of terrestrial gravity. Analyses included assessments of final cell count, size, morphology, and soluble metal concentrations. Under Earth gravity, with the addition of Fe 3+ and H 2 /CO 2 , A. ferrooxidans grew in the presence of regolith simulants to a final cell density comparable to controls without regoliths. The simulated microgravity appeared to enable cells to grow to a higher cell density in the presence of lunar regolith simulants. Clinostat cultures of A. ferrooxidans solubilised higher amounts of Si, Mn and Mg from lunar and Martian regolith simulants than abiotic controls. Electron microscopy observations revealed that microgravity stimulated the biosynthesis of intracellular nanoparticles (most likely magnetite) in anaerobically grown A. ferrooxidans cells. These results suggested that A. ferrooxidans has the potential for metal bioleaching and the production of useful nanoparticles in space.

With the development of transcriptomic technologies, we are able to quantify precise changes in gene expression profiles from astronauts and other organisms exposed to spaceflight. Members of NASA GeneLab and GeneLab-associated analysis working groups (AWGs) have developed a consensus pipeline for analyzing short-read RNA-sequencing data from spaceflight-associated experiments. The pipeline includes quality control, read trimming, mapping, and gene quantification steps, culminating in the detection of differentially expressed genes. This data analysis pipeline and the results of its execution using data submitted to GeneLab are now all publicly available through the GeneLab database. We present here the full details and rationale for the construction of this pipeline in order to promote transparency, reproducibility, and reusability of pipeline data; to provide a template for data processing of future spaceflight-relevant datasets; and to encourage cross-analysis of data from other databases with the data available in GeneLab., Competing Interests: The authors declare no competing interests.

Biofilms, surface-adherent microbial communities, are associated with microbial fouling and corrosion in terrestrial water-distribution systems. Biofilms are also present in human spaceflight, particularly in the Water Recovery System (WRS) on the International Space Station (ISS). The WRS is comprised of the Urine Processor Assembly (UPA) and the Water Processor Assembly (WPA) which together recycles wastewater from human urine and recovered humidity from the ISS atmosphere. These wastewaters and various process streams are continually inoculated with microorganisms primarily arising from the space crew microbiome. Biofilm-related fouling has been encountered and addressed in spacecraft in low Earth orbit, including ISS and the Russian Mir Space Station. However, planned future missions beyond low Earth orbit to the Moon and Mars present additional challenges, as resupplying spare parts or support materials would be impractical and the mission timeline would be in the order of years in the case of a mission to Mars. In addition, future missions are expected to include a period of dormancy in which the WRS would be unused for an extended duration. The concepts developed in this review arose from a workshop including NASA personnel and representatives with biofilm expertise from a wide range of industrial and academic backgrounds. Here, we address current strategies that are employed on Earth for biofilm control, including antifouling coatings and biocides and mechanisms for mitigating biofilm growth and damage. These ideas are presented in the context of their applicability to spaceflight and identify proposed new topics of biofilm control that need to be addressed in order to facilitate future extended, crewed, spaceflight missions., (© 2020 The Authors.)

In search of pharmaceutically active products to control type 2 diabetes, five brown seaweeds ( Silvetia compressa , Cystoseira osmundacea , Ecklonia arborea , Pterygophora californica , and Egregia menziesii ) from the Northwest Mexican Pacific coast were investigated. Proximate composition and total polyphenol content (TPC) as phloroglucinol equivalents (PGE) were determined for the five seaweed powders and their respective hydroethanolic (1 : 1) extracts. Extracts were screened for their radical scavenging activity (DPPH and ORAC) and glycosidase inhibitory activity. HPLC-DAD, HPLC-MS-TOF, and ATR-FT-IR methodologies were used to identify the most abundant phlorotannins and sulfated polysaccharides in the extracts. Hydroethanolic extracts contained minerals (17 to 59% of the dry matter), proteins (4 to 9%), ethanol-insoluble polysaccharides (5.4 to 53%), nitrogen-free extract (NFE) (24.4 to 70.1%), lipids (5 to 12%), and TPC (2.6 to 47.7 g PGE per 100 g dry extract). S. compressa and E. arborea dry extracts presented the lowest ash content (26 and 17%, respectively) and had some of the highest phenolic (47.7 and 15.2 g PGE per 100 g extract), NFE (57.3 and 70.1%), and soluble polysaccharide (19.7 and 53%) contents. S. compressa and E. arborea extracts had the highest antioxidant activity (IC 50 DPPH 1.7 and 3.7 mg mL -1 ; ORAC 0.817 and 0.801 mmol Trolox equivalent/g extract) and the highest α -amylase and α -glucosidase inhibitory capacities (IC 50 940 and 1152  μ g mL -1 against α -amylase and 194 and 647  μ g mL -1 against α -glucosidase). The most abundant phlorotannins identified in the extracts were phloretol, fucophloroethol, and two- and three-phloroglucinol unit (PGU) phlorotannins. Laminarin, fucoidan, and alginate were among the sulfated polysaccharides identified in the extracts. The bioactivities of S. compressa and E. arborea extracts were mainly related with their contents of three PGU phlorotannins and sulfated polysaccharides (e.g., fucoidan, laminarin, and alginate). These results suggest S. compressa and E. arborea are potential candidates for food products and nutraceutical and pharmaceutical preparations, and as additives for diabetes management., Competing Interests: The authors declare that there are no conflicts of interest., (Copyright © 2019 Ana Rocío Múzquiz de la Garza et al.)

A dynamic headspace sorptive extraction (DHS) combined with thermal desorption (TD) and coupled with gas chromatography-mass spectrometry (GC/MS) was developed for the determination of forty-four volatile aroma metabolites (VAMs) which describe aroma fingerprints of wines. The response surface methodology (RSM) and a central composite design (CCD) was used to obtain the optimal values for the experimental extraction variables, and the results were assessed by an analysis of variance (ANOVA) and a principal component analysis (PCA). The VAMs exhibited optimal extraction with the high levels of salt concentration (1.5 g NaCl) and using an extraction temperature of 40 °C during 10 min, and a subsequent purge volume of 300 mL. Subsequently, the calibration curves were created for the quantification of each VAMs with seven levels of concentration obtaining a correlation coefficients (R 2 ) above 0.9900 for all of them. The proposed method was successfully validated and showed good precision and accuracy values for all VAMs. Lastly, the method was applied to quantify VAMs, responsible for the aroma fingerprints of white and red wines, from different Denomination of Origin., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Power ultrasound is applied in food technology to intensify extraction processes, due to the phenomena ultrasonic energy induces in the medium, enhancing mass transfer. The purpose of this work was the acoustic characterization of four transducers of different geometries and the evaluation of their performance in the ultrasonically assisted supercritical fluid extraction of antioxidants from oregano. The transducers differed in the amount of energy transmitted into the medium. Designs varied from the base model (T1), a larger cylindrical headmass (T2), a stepped circular section sonotrode (T3) and a multiplate configuration (T4). The highest nominal power density provided according to the calorimetric method was for T4 (151.6 ± 7.1 W/L). The T2 produced a more uniform acoustic field and a higher acoustic pressure (150.6 ± 20.5 kPa). Both parameters had an impact on total phenolics and antioxidants extraction with CO 2 under supercritical conditions (35 MPa, 35 °C, 2.3% ethanol as co-solvent). T4 and T2 were equally efficient (4.0 ± 0.2 and 4.2 ± 0.2 mg GA/g) for phenolic extraction, and with respect to antioxidant capacity, the best performance was that of T4 (26.4 ± 1.1 μmol TE/g). Of the antioxidant compounds extracted, flavones and flavanones were identified. Therefore, transducer geometry influenced the amount and distribution of energy transmitted into the medium, thus determining the efficiency of the extraction process., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Biofilm growth has been observed in Soviet/Russian (Salyuts and Mir), American (Skylab), and International (ISS) Space Stations, sometimes jeopardizing key equipment like spacesuits, water recycling units, radiators, and navigation windows. Biofilm formation also increases the risk of human illnesses and therefore needs to be well understood to enable safe, long-duration, human space missions. Here, the design of a NASA-supported biofilm in space project is reported. This new project aims to characterize biofilm inside the International Space Station in a controlled fashion, assessing changes in mass, thickness, and morphology. The space-based experiment also aims at elucidating the biomechanical and transcriptomic mechanisms involved in the formation of a "column-and-canopy" biofilm architecture that has previously been observed in space. To search for potential solutions, different materials and surface topologies will be used as the substrata for microbial growth. The adhesion of bacteria to surfaces and therefore the initial biofilm formation is strongly governed by topographical surface features of about the bacterial scale. Thus, using Direct Laser-Interference Patterning, some material coupons will have surface patterns with periodicities equal, above or below the size of bacteria. Additionally, a novel lubricant-impregnated surface will be assessed for potential Earth and spaceflight anti-biofilm applications. This paper describes the current experiment design including microbial strains and substrata materials and nanotopographies being considered, constraints and limitations that arise from performing experiments in space, and the next steps needed to mature the design to be spaceflight-ready.

Bacteria grown in space experiments under microgravity conditions have been found to undergo unique physiological responses, ranging from modified cell morphology and growth dynamics to a putative increased tolerance to antibiotics. A common theory for this behavior is the loss of gravity-driven convection processes in the orbital environment, resulting in both reduction of extracellular nutrient availability and the accumulation of bacterial byproducts near the cell. To further characterize the responses, this study investigated the transcriptomic response of Escherichia coli to both microgravity and antibiotic concentration. E. coli was grown aboard International Space Station in the presence of increasing concentrations of the antibiotic gentamicin with identical ground controls conducted on Earth. Here we show that within 49 h of being cultured, E. coli adapted to grow at higher antibiotic concentrations in space compared to Earth, and demonstrated consistent changes in expression of 63 genes in response to an increase in drug concentration in both environments, including specific responses related to oxidative stress and starvation response. Additionally, we find 50 stress-response genes upregulated in response to the microgravity when compared directly to the equivalent concentration in the ground control. We conclude that the increased antibiotic tolerance in microgravity may be attributed not only to diminished transport processes, but also to a resultant antibiotic cross-resistance response conferred by an overlapping effect of stress response genes. Our data suggest that direct stresses of nutrient starvation and acid-shock conveyed by the microgravity environment can incidentally upregulate stress response pathways related to antibiotic stress and in doing so contribute to the increased antibiotic stress tolerance observed for bacteria in space experiments. These results provide insights into the ability of bacteria to adapt under extreme stress conditions and potential strategies to prevent antimicrobial-resistance in space and on Earth.

Bacteria will accompany humans in our exploration of space, making it of importance to study their adaptation to the microgravity environment. To investigate potential phenotypic changes for bacteria grown in space, Escherichia coli was cultured onboard the International Space Station with matched controls on Earth. Samples were challenged with different concentrations of gentamicin sulfate to study the role of drug concentration on the dependent variables in the space environment. Analyses included assessments of final cell count, cell size, cell envelope thickness, cell ultrastructure, and culture morphology. A 13-fold increase in final cell count was observed in space with respect to the ground controls and the space flight cells were able to grow in the presence of normally inhibitory levels of gentamicin sulfate. Contrast light microscopy and focused ion beam/scanning electron microscopy showed that, on average, cells in space were 37% of the volume of their matched controls, which may alter the rate of molecule-cell interactions in a diffusion-limited mass transport regime as is expected to occur in microgravity. TEM imagery showed an increase in cell envelope thickness of between 25 and 43% in space with respect to the Earth control group. Outer membrane vesicles were observed on the spaceflight samples, but not on the Earth cultures. While E. coli suspension cultures on Earth were homogenously distributed throughout the liquid medium, in space they tended to form a cluster, leaving the surrounding medium visibly clear of cells. This cell aggregation behavior may be associated with enhanced biofilm formation observed in other spaceflight experiments.

Bacteria behave differently in space, as indicated by reports of reduced lag phase, higher final cell counts, enhanced biofilm formation, increased virulence, and reduced susceptibility to antibiotics. These phenomena are theorized, at least in part, to result from reduced mass transport in the local extracellular environment, where movement of molecules consumed and excreted by the cell is limited to diffusion in the absence of gravity-dependent convection. However, to date neither empirical nor computational approaches have been able to provide sufficient evidence to confirm this explanation. Molecular genetic analysis findings, conducted as part of a recent spaceflight investigation, support the proposed model. This investigation indicated an overexpression of genes associated with starvation, the search for alternative energy sources, increased metabolism, enhanced acetate production, and other systematic responses to acidity-all of which can be associated with reduced extracellular mass transport., Competing Interests: The authors have declared that no competing interests exist.

Concentrated agave sap (CAS) has gained popularity as an unrefined sweetener. It is obtained by boiling "aguamiel" that contains phytochemicals with diverse bioactivities. Saponins have been the most widely studied agave phytochemicals due to their cancer antiproliferative effect but their concentration may vary due to maturity of the agave plant and collection site. In this study, 18 CAS samples produced in different states of Mexico were analyzed using multivariate methods to determine which physicochemical or phytochemical parameters were responsible for variation. Additionally, extracts with different saponin profiles were tested to determine possible correlations with antiproliferative activity. Total soluble solids, pH, and water activity were similar to those reported for other agave sweeteners. Antioxidant capacity of samples was correlated to browning index. Eleven steroidal saponins were found in CAS samples and they were the main source of variability. Magueyoside B, a kammogenin tetraglycoside, was the most abundant saponin in all samples. With respect to bioactivity, multivariate analysis indicated that magueyoside B and a gentrogenin tetraglycoside were compounds strongly related with bioactivity. CAS from Hidalgo, Puebla, and Veracruz had higher concentration of magueyoside B than from the other kamogenin tetraglycoside found in the samples from other Mexican states. These results could be used as a first approach to characterize and standardize CAS to validate the potential health benefits derived from its consumption., (© 2016 Institute of Food Technologists®)

Separation of potentially bioactive components from foods and plant extracts is one of the main challenges for their study. Centrifugal partition chromatography has been a successful technique for the screening and identification of molecules with bioactive potential, such as steroidal saponins. Agave is a source of steroidal saponins with anticancer potential, though the activity of these compounds in concentrated agave sap has not been yet explored. In this study, fast centrifugal partition chromatography (FCPC) was used coupled with in vitro tests on HT-29 cells as a screening procedure to identify apoptotic saponins from an acetonic extract of concentrated agave sap. The three most bioactive fractions obtained by FCPC at partition coefficients between 0.23 and 0.4 contained steroidal saponins, predominantly magueyoside b. Flow cytometry analysis determined that the fraction rich in kammogenin and manogenin glycosides induced apoptosis, but when gentrogenin and hecogenin glycosides were also found in the fraction, a necrotic effect was observed. In conclusion, this study provides the evidence that steroidal saponins in concentrated agave sap were potential inductors of apoptosis and that it was possible to separate them using fast centrifugal partition chromatography.

Steroidal saponins have shown beneficial health effects. Agave spp. leaves and rhizomes are sources of these compounds, but their presence has not been reported in the aguamiel. Aguamiel is the sweet edible sap from mature agave, and its quality is influenced by the plant ripening stage. The purpose of this research was to identify and quantitate saponins in aguamiel from Agave americana and Agave salmiana at two ripening stages. Saponins and sapogenins were identified with HPLC/ESI-MS/TOF and quantitated with HPLC/ELSD. Results proved the presence of saponins derived from kammogenin, manogenin, gentrogenin, and hecogenin. The saponin content in aguamiel from immature A. salmiana was 2-fold higher (478.3 protodioscin equivalents (PE) μg/g aguamiel (DM)) compared with A. americana (179.0 PE μg/g aguamiel (DM)). In both species, saponin content decreased when plants reached sexual maturity. This should be considered before evaluating the effects of Agave spp. as a source of bioactive saponins.

In this work, the morphological and chemical properties of Chardonnay and Gewürztraminer aromatic grapes (northern Spain) have been studied with the aim to assess their response to chamber-drying under controlled conditions and compare it with that of Pedro Ximenez grapes (southern Spain). Morphological characteristics, such as weight, size and roundness, and other of the skin such as thickness, enabled discrimination of the two types of grapes varieties. Changes in browning index, colour, antioxidant activity, aroma compounds determined by GC-MS and flavan-3-ols and flavonols concentrations determined by HPLC-DAD were studied during drying. Based on the results, Chardonnay and Gewürztraminer grapes contained increased amounts of flavan-3-ol derivatives, which are the greatest contributors to polymerization and condensation reactions. Also, their smaller size resulted in faster drying and leads to sugary musts that were lighter-coloured, less brown and more aromatic than Pedro Ximenez grapes., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A dynamic headspace sorptive extraction (DHS) combined with thermal desorption (TD) and coupled with gas chromatography-mass spectrometry (GC/MS) was developed for the determination of 11 esters which contribute to the fruity aroma in sweet wines. A full factorial (4 factors, 2 level) experiment design was used to optimize the extraction conditions and the results were evaluated by multiple linear regression (MLR) and principal component analysis (PCA). The esters showed optimal extraction using an extraction temperature of 30°C during 20 min, and a subsequent purge volume of 300 mL and dry volume of 50 mL. Afterwards, quantification was achieved using calibration curves constructed for each ester with linear regression equations having correlation coefficients (R(2)) ranging from 0.9894 to 0.9981. The proposed method was successfully validated and showed good intermediate precision, repeatability and accuracy values for all the monitored compounds. Finally, the method was applied to quantify esters, with fruity aromatic notes, of sweet white and red wines, elaborated with different winemaking processes., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The phenolic, flavonoid, and antioxidant contents of methanol extracts of nine samples of Mexican cactus ( Opuntia spp.) cladodes processed into flours were studied. Opuntia undulata contained the highest amount of phenols [905.08 ± 64.51 μg of gallic acid equivalents (GAE)/g]. The oxygen radical absorbance capacity (ORAC) of the cladode flour extracts indicated that Opuntia robusta var. Gavia [738.8 ± 89.9 μmol of Trolox equivalents (TE)/g] contained the highest antioxidant capacity. ORAC values significantly correlated to total phenols but not to flavonoid contents and were comparable to cranberries and blackberries. Glycosidic forms of isorhamnetin and kaempferol were identified via high-performance liquid chromatography-photodiode array (HPLC-PDA) and HPLC-mass spectrometry (MS), with isorhamnetin being the most abundant flavonol in all samples, except for Opuntia lindheimeri . The effectiveness of acid hydrolysis varied among species because of the different flavonol profiles. For some varieties, the triglycosidic forms were partially acid-hydrolyzed, giving an increase in the content of diglycosides. Optimization of hydrolysis for each species is required to estimate the total amount of each flavonol.

Odor compounds in Amontillado sherry white wine obtained by means of biological aging first and oxidative aging second in American oak casks were determined by gas chromatography-olfactometry. Sniffing revealed fruity, fatty, chemical, spicy, vegetable, floral and empyreumatic odors, the first being the most common. Olfactometric intensity was assessed on a four-point scale. Most changes were detected during the first years of the oxidative aging step. Ethyl isobutanoate, ethyl butanoate, ethyl octanoate, and eugenol were the strongest odor compounds detected by sniffing in wines. The odor spectrum values for all active odorants were calculated in relation to ethyl octanoate, this compound being the most potent odorant. On the basis of olfactometric intensities and odor spectrum values, ethyl octanoate, ethyl butanoate, eugenol, ethyl isobutanoate, and sotolon can be deemed the main group of potent odorants in Amontillado wines. These compounds maintained similar relative contributions to the aroma profile during the oxidative aging step.

The aroma of young and biologically aged sherry wines from Pedro Ximenez grape cultivated conventionally and ecologically has been studied. Fifty-five compounds were quantified by GC, and the odor activity values for the 19 odor-active compounds considered were grouped into 8 odorant series, the fruity and fatty series showing the highest OAVs. The OAVs of the eight series were subjected to a principal component analysis. PC1 separated the young wines from the aged wines, also distinguishing the traditional young wines from the ecological young ones, whereas PC2 was effective only in separating the traditional aged Fino wines from the ecologically aged ones. The ecological Fino wines showed lower values than traditional Fino wines for the OAVs of all the series, except for the balsamic and fatty series, the ecologically aged wines showing a sensorial profile similar to that of the traditional Fino but with a lower odor intensity.

Pedro Ximenez sweet wines obtained following the typical criaderas and solera method for sherry wines and subjected to oxidative aging for 0, 1.3, 4.2, 7.0, or 11.5 years were studied in terms of color and aroma fraction by using the CIELab method and gas chromatography, respectively. The parameters defining the CIELab color space (a*, b*, and L*) were subjected to a multiple-range test (p<0.05) that allowed discrimination of the five wine aging levels studied into five uniform groups according to aging time. The aroma fraction was found to include 15 active odorant compounds with OAV > 1 that enriched the wines with fruity, fatty, floral, and balsamic notes during the aging process. The changes in color parameters and active odorants were not linearly related to aging time, being especially marked during the first 1.3 years and then less substantial up to the 7 years, the oldest wines exhibiting sensorial properties markedly departing from all others. For the wines aged over 1.3 years (minimum aging), 2,3-butanedione, linalool, and decanal can be used as reliable fingerprints of the older wines' quality.

The odor activity values (OAVs) for 49 aroma compounds in commercial sherry pale white wines were grouped, according to the similarity of their aroma descriptors, into nine odor classes with a view to establishing the aroma profile for this type of wine. The results revealed the profile to be largely comprised of the series named "fruity" and "balsamic", mainly as a result of the 1,1-diethoxyethane content in the wines. The same series were calculated from the OAVs obtained in biological aging experiments, carried out with selected strains of the flor yeasts Saccharomyces cerevisiae and Saccharomyces bayanus, over a period of 9 months. Based on the aroma profiles thus obtained, after 6 months of aging the latter race yielded OAVs for the fruity and balsamic series not significantly different (p < 0.05) from those for commercial wines aged for 5 years. However, except for the series named "solvent", all others exhibited lower values in the experiments carried out with selected strains than in the commercial wines, mainly as a result of the absence of contact with wood of the former wines. Taking into account the results, the biological aging of this type of sherry wine can be shortened by subjecting it to controlled aging with selected yeast strains in a first stage and subsequently allowing it to stand in wood casks in a second stage.

The effect of an acceleration assay, carried out with a periodic aeration and an increased surface/volume ratio, on various aroma compounds of "fino" Sherry wines aging under a veil of a pure culture of Saccharomyces cerevisiae race capensis G1 flor film yeast was studied. The results were subjected to multifactor analysis of variance, and the compounds simultaneously depending on acceleration conditions and aging time at p < 0.01 were subjected to principal component analysis. The first component, accounting for 86.14% of the overall variance, was mainly defined by acetaldehyde and its derivatives 1,1-diethoxyethane and acetoin. These compounds reached higher concentrations in accelerated aging wines in a shorter time than they did in control wines, and no browning problems were detected. Taking into account that these compounds can be used as indicators for biological aging of "fino" Sherry wines, the acceleration condition assayed can be applied to shorten the time of this process.

At least three independent groups of natriuretic hormones have been isolated over the past ten years. Two, atrial natriuretic factor (ANF) and brain natriuretic peptide (BNP), are proteins and the third is made up of digitalis-like substances (DLS). The present report concerns the isolation, substantial purification and biologic actions of an entirely different natriuretic hormone (NH) which appears to be steroidal in nature and an isomer of cortisone. The source of NH was uremic urine. Purification involved successive chromatographic steps including gel filtration and multiple HPLC runs through C-18 resins. A translucent crystal ultimately was obtained. The product was examined using mass spectroscopy with trimethylsilyl derivatization. Only one compound was identifiable. The characteristics of the molecule include: a molecular weight, 360.4; a molecular formula, C21H28O5; a steroidal nucleus; UV absorption at 220 and 290 nm; and intrinsic fluorescence. The onset of action occurs within minutes both in the rat and, as previously shown, in several in vitro systems including the frog skin, toad bladder, fibroblasts and renal tubular epithelial cells grown in culture and isolated perfused cortical collecting tubules. In contrast to DLS, NH has been previously shown not to cross react with digoxin antibodies. Moreover, when given to intact rats, it produces a profound natriuresis but little or no kaliuresis. In contrast to ANF and BNP the compound is active orally as well as intravenously. It is clearly different from cortisone, based both on its biologic and mass spectroscopic characteristics.