-
Journal of Visualized Experiments : JoVE Jul 2019We describe a rapid tissue donation program for individuals with multiple sclerosis (MS) that requires scientists and technicians to be on-call 24/7, 365 days a year....
We describe a rapid tissue donation program for individuals with multiple sclerosis (MS) that requires scientists and technicians to be on-call 24/7, 365 days a year. Participants consent to donate their brain and spinal cord. Most patients were followed by neurologists at the Cleveland Clinic Mellen Center for MS Treatment and Research. Their clinical courses and neurological disabilities are well-characterized. Soon after death, the body is transported to the MS Imaging Center, where the brain is scanned in situ by 3 T magnetic resonance imaging (MRI). The body is then transferred to the autopsy room, where the brain and spinal cord are removed. The brain is divided into two hemispheres. One hemisphere is immediately placed in a slicing box and alternate 1 cm-thick slices are either fixed in 4% paraformaldehyde for two days or rapidly frozen in dry ice and 2-methylbutane. The short-fixed brain slices are stored in a cryopreservation solution and used for histological analyses and immunocytochemical detection of sensitive antigens. Frozen slices are stored at -80 °C and used for molecular, immunocytochemical, and in situ hybridization/RNA scope studies. The other hemisphere is placed in 4% paraformaldehyde for several months, placed in the slicing box, re-scanned in the 3 T magnetic resonance (MR) scanner and sliced into centimeter-thick slices. Postmortem in situ MR images (MRIs) are co-registered with 1 cm-thick brain slices to facilitate MRI-pathology correlations. All brain slices are photographed and brain white-matter lesions are identified. The spinal cord is cut into 2 cm segments. Alternate segments are fixed in 4% paraformaldehyde or rapidly frozen. The rapid procurement of postmortem MS tissues allows pathological and molecular analyses of MS brains and spinal cords and pathological correlations of brain MRI abnormalities. The quality of these rapidly-processed postmortem tissues (usually within 6 h of death) is of great value to MS research and has resulted in many high-impact discoveries.
Topics: Autopsy; Brain; Humans; Magnetic Resonance Imaging; Multiple Sclerosis; Spinal Cord; White Matter
PubMed: 31380830
DOI: 10.3791/59511 -
Mycologia 2021In an era of rapid climate change and expansion of desertification, the extremely harsh conditions of drylands are a true challenge for microbial life. Under drought...
In an era of rapid climate change and expansion of desertification, the extremely harsh conditions of drylands are a true challenge for microbial life. Under drought conditions, where most life forms cannot survive, rocks represent the main refuge for life. Indeed, the endolithic habitat provides thermal buffering, physical stability, and protection against incident ultraviolet (UV) radiation and solar radiation and, to some extent, ensures water retention to microorganisms. The study of these highly specialized extreme-tolerant and extremophiles may provide tools for understanding microbial interactions and processes that allow them to keep their metabolic machinery active under conditions of dryness and oligotrophy that are typically incompatible with active life, up to the dry limits for life. Despite lithobiontic communities being studied all over the world, a comprehensive understanding of their ecology, evolution, and adaptation is still nascent. Herein, we survey the fungal component of these microbial ecosystems. We first provide an overview of the main defined groups (i.e., lichen-forming fungi, black fungi, and yeasts) of the most known and studied Antarctic endolithic communities that are almost the only life forms ensuring ecosystem functionality in the ice-free areas of the continent. For each group, we discuss their main traits and their diversity. Then, we focus on the fungal taxonomy and ecology of other worldwide endolithic communities. Finally, we highlight the utmost importance of a global rock survey in order to have a comprehensive view of the diversity, distribution, and functionality of these fungi in drylands, to obtain tools in desert area management, and as early alarm systems to climate change.
Topics: Adaptation, Physiological; Antarctic Regions; Biodiversity; Climate Change; Desert Climate; Droughts; Extreme Environments; Fungi; Geologic Sediments; Lichens
PubMed: 33232202
DOI: 10.1080/00275514.2020.1816761 -
Journal of Proteome Research Sep 2021Synthetic peptides are a critical requirement for the development and application of targeted mass spectrometry (MS)-based assays for the quantitation of proteins from...
Synthetic peptides are a critical requirement for the development and application of targeted mass spectrometry (MS)-based assays for the quantitation of proteins from biological matrices. Transporting synthetic peptides on dry ice from one laboratory to another is costly and often difficult because of country-specific import and export regulations. Therefore, in this study, we assessed the impact of leaving a lyophilized mixture consisting of 125 peptides at room temperature for up to 20 days, and we assessed the effect on the quantitative performance of multiple reaction monitoring-MS (MRM-MS) assays. The findings suggest that there are no significant differences in the MRM-MS results for the time points assessed in this study (up to 20 days). All the calibration curves and quality control (QC) samples met the acceptance criteria for precision and accuracy (raw data are available via the public MS data repository PanoramaWeb, identifier: /MRM Proteomics/2020_BAK125_RT). The number of endogenous proteins quantifiable across five plasma samples was consistently between 87 and 99 out of 125 for all time points. Moreover, the coefficients of variation (CVs) calculated for the majority of peptide concentrations across all samples and time points were <5%. In addition, a lyophilized peptide mixture was transported from Canada to Iceland without dry ice. The results showed that there was no significant difference in the quantitative performance, with the determined concentrations of most proteins in the samples falling within 30% between the analyses performed on the same three plasma samples in Iceland and those in Canada. Overall, a comparison of the results obtained in Canada and in Iceland indicated that the peptides were stable under the conditions tested and also indicated that shipping lyophilized peptide mixtures without dry ice, but in the presence of sufficient desiccant material, could be a feasible option in cases where transport difficulties may arise or dry-ice sublimation may occur.
Topics: Humans; Mass Spectrometry; Peptides; Proteins; Proteomics; Temperature
PubMed: 34270269
DOI: 10.1021/acs.jproteome.1c00249 -
The Journal of the Acoustical Society... Dec 2022The ice-covered Arctic Ocean constitutes a unique underwater acoustic waveguide; it is a half-channel, upward refracting environment possessing a rough upper boundary...
The ice-covered Arctic Ocean constitutes a unique underwater acoustic waveguide; it is a half-channel, upward refracting environment possessing a rough upper boundary consisting of sea ice of varying thickness. The sea ice itself is an acoustic waveguide, capable of supporting the propagation of compressional and shear waves. In particular, the ice supports compressional wave resonances created by impulsive forces on the upper surface of the ice. During ICEX20 and ICEX22, observations were made of compressional wave resonances excited by hammer drops, as well as by near-impulsive signals generated from the compression of dry snow underfoot while walking on the ice. Results demonstrate that ice thickness can be inferred from compressional wave resonances in the sea ice waveguide using signals generated by walking on the snow-covered ice. Inferred ice thickness estimates were consistent with observations made by magnetic induction and physical measurements in holes drilled through the ice. Average first- and multi-year ice thicknesses were inferred to be 1.1-1.3 m and 2.4-2.5 m, respectively.
PubMed: 36586831
DOI: 10.1121/10.0016632 -
Biology Jan 2023Because of climate change, the McMurdo Dry Valleys of Antarctica (MCM) have experienced an increase in the frequency and magnitude of summer pulse warming and surface...
Because of climate change, the McMurdo Dry Valleys of Antarctica (MCM) have experienced an increase in the frequency and magnitude of summer pulse warming and surface ice and snow melting events. In response to these environmental changes, some nematode species in the MCM have experienced steady population declines over the last three decades, but , a mesophilic nematode species, has responded with a steady increase in range and abundance. To determine how responds to increasing temperatures, we measured metabolic heat and CO production rates and calculated O consumption rates as a function of temperature at 5 °C intervals from 5 to 50 °C. Heat, CO production, and O consumption rates increase approximately exponentially up to 40 °C, a temperature never experienced in their polar habitat. Metabolic rates decline rapidly above 40 °C and are irreversibly lost at 50 °C due to thermal stress and mortality. , a much more widespread nematode that is found in more temperate environments reaches peak metabolic heat rate at just 27 °C, above which it experiences high mortality due to thermal stress. At temperatures from 10 to 40 °C, produces about 6 times more CO than the O it consumes, a respiratory quotient indicative of either acetogenesis or de novo lipogenesis. No potential acetogenic microbes were identified in the microbiome, suggesting that is producing increased CO as a byproduct of de novo lipogenesis. This phenomenon, in conjunction with increased summer temperatures in their polar habitat, will likely lead to increased demand for carbon and subsequent increases in CO production, population abundance, and range expansion. If such changes are not concomitant with increased carbon inputs, we predict the MCM soil ecosystems will experience dramatic declines in functional and taxonomic diversity.
PubMed: 36671801
DOI: 10.3390/biology12010109 -
Journal of Environmental Management Sep 2022A cryogel solid amine adsorbent with a laminated structure has been prepared by crosslinking polyethylenimine (PEI) with ethylene glycol diglycidyl ether (EGDE) at a low...
A cryogel solid amine adsorbent with a laminated structure has been prepared by crosslinking polyethylenimine (PEI) with ethylene glycol diglycidyl ether (EGDE) at a low temperature via liquid nitrogen treatment and freeze-drying. The effects of cryogenic treatment on the morphology of the cryogels were investigated. The liquid nitrogen treatment and freeze drying were critical to create the layered structure. The fast formation of ice crystals at 77 K served as a template which directed the ordered lamellar structure of the PEI and EGDE cross-linked polymer networks. The PEI cryogel adsorbent showed excellent CO adsorption performance both in dry and wet conditions. In dry conditions, the PEI-gel-5-0.25 cryogel showed a 5.60 mmol/g of CO adsorption capacity at 75 °C. After being swelled with water, the PEI-gel-15-0.25 cryogel showed an extremely high CO adsorption capacity of 11.39 mmol/g at 25 °C. The adsorption behaviors of adsorbents with varied water contents were explained using kinetic simulations and intraparticle diffusion simulations. It was found that the presence of water can significantly enhance the diffusion process. The regeneration performance was examined in both dry and wet conditions. After 20 adsorption-desorption cycles, the adsorption capacity of the regenerated PEI cryogel had barely decreased, indicating reliable regeneration stability.
Topics: Adsorption; Carbon; Carbon Dioxide; Cryogels; Nitrogen; Polyethyleneimine; Water
PubMed: 35653848
DOI: 10.1016/j.jenvman.2022.115400 -
Health Science Reports Sep 2023Winter endurance athletes have a high prevalence of exercise-induced bronchoconstriction (EIB) and asthma, probably due to repeated and prolonged inhalation of cold and...
BACKGROUND AND AIMS
Winter endurance athletes have a high prevalence of exercise-induced bronchoconstriction (EIB) and asthma, probably due to repeated and prolonged inhalation of cold and dry air. Heat- and moisture-exchanging devices (HME) warm and humidify inhaled air and prevent EIB. The aim of this study was to share cross-country skiers and biathletes' experiences of training and competition in low temperatures, views on temperature limits, usage of HME, and consequences of cold exposure on their health.
METHODS
Eleven Swedish World Championship or Olympic medalists in cross-country skiing and biathlon were interviewed and transcripts were analyzed using qualitative content analysis.
RESULTS
Participants described how cold temperatures predominantly affected the airways, face, and extremities. During training, extreme cold was managed by choosing warmer clothing, modification of planned sessions, use of HME, delaying training, or changing location. In competition, participants described limited possibility for such choices and would prefer adjustment of existing rules (i.e., more conservative temperature limits), especially since they understood elite skiing in low temperatures to present an occupational hazard to their health. Participants had at times used HMEs during training in cold environments but described mixed motives for their use-that HMEs warm and humidify cold inhaled air but introduce additional resistance to breathing and can cause problems due to mucus and ice build-up. Skiers also perceived that they had become more sensitive to cold during the latter part of their careers.
CONCLUSIONS
The present study gives a unique insight into the "cold" reality of being an elite athlete in skiing and biathlon. Cold exposure results in negative health consequences that are preventable, which means that rules must be followed, and organizers should acknowledge responsibility in protecting athletes from occupational hazards. Development of evidence-based guidelines for protection of athletes' respiratory health should be a focus for future translational research.
PubMed: 37662540
DOI: 10.1002/hsr2.1511 -
Journal of Pain and Symptom Management Nov 2023Thirst and xerostomia are significant and highly distressing symptoms experienced by patients receiving palliative and end-of-life care.
CONTEXT
Thirst and xerostomia are significant and highly distressing symptoms experienced by patients receiving palliative and end-of-life care.
OBJECTIVES
Determine a reduction of thirst intensity and perceptions of dry mouth on a numerical scale following both the experimental intervention (mini mint ice cubes) and control (plain ice chips).
METHODS
Cross-over Randomized Controlled Trial (RCT) to assess the effectiveness of novel intervention in the treatment of dry mouth and the sensation of thirst in palliative care patients.
RESULTS
Patients rated the severity of their symptoms of dry mouth and thirst using a numeric rating scale (NRS). On commencing the study and preintervention, all patients suffered severe dry mouth and thirst (≥5/10). Mint and plain ice cubes produced improvement of symptoms immediately after interventions. Results from dry mouth ratings show, a decrease of 1.6 points for plain ice cubes (P < 0.0001), on average, ratings for mint ice cubes decreased 3.7 (P < 0.0001). For the sensation of thirst, the plain ice cube intervention group rating decreased 1.7 points (P < 0.006), ratings for mint ice cubes decreased 3.4 points (P < 0.0001). The average decrease in dry mouth and thirst intensity scores from preintervention to postintervention were significantly greater for mint ice cubes (P < 0.05) and 86.6% of patients preferred mint ice cubes.
CONCLUSION
This trial found that while usual mouth care and the intervention were both able to reduce the intensity of dry mouth and the sensation of thirst, the mint intervention had a greater response.
PubMed: 37562697
DOI: 10.1016/j.jpainsymman.2023.08.005 -
Environmental Microbiology Dec 2022The McMurdo Dry Valleys (MDVs), Antarctica, represent a cold, desert ecosystem poised on the threshold of melting and freezing water. The MDVs have experienced dramatic...
The McMurdo Dry Valleys (MDVs), Antarctica, represent a cold, desert ecosystem poised on the threshold of melting and freezing water. The MDVs have experienced dramatic signs of climatic change, most notably a warm austral summer in 2001-2002 that caused widespread flooding, partial ice cover loss and lake level rise. To understand the impact of these climatic disturbances on lake microbial communities, we simulated lake level rise and ice-cover loss by transplanting dialysis-bagged communities from selected depths to other locations in the water column or to an open water perimeter moat. Bacteria and eukaryote communities residing in the surface waters (5 m) exhibited shifts in community composition when exposed to either disturbance, while microbial communities from below the surface were largely unaffected by the transplant. We also observed an accumulation of labile dissolved organic carbon in the transplanted surface communities. In addition, there were taxa-specific sensitivities: cryptophytes and Actinobacteria were highly sensitive particularly to the moat transplant, while chlorophytes and several bacterial taxa increased in relative abundance or were unaffected. Our results reveal that future climate-driven disturbances will likely undermine the stability and productivity of MDV lake phytoplankton and bacterial communities in the surface waters of this extreme environment.
Topics: Lakes; Phytoplankton; Ecosystem; Antarctic Regions; Bacteria; Water
PubMed: 35860854
DOI: 10.1111/1462-2920.16113 -
Stem Cell Research & Therapy Mar 2021Neural stem cell (NSC) therapy remains one of the most potential approaches for the treatment of neurological disorders. The discovery of human induced pluripotent stem...
BACKGROUND
Neural stem cell (NSC) therapy remains one of the most potential approaches for the treatment of neurological disorders. The discovery of human induced pluripotent stem cells (hiPSCs) and the establishment of hiPSC-derived human neural stem cells (hiNSCs) have revolutionized the technique of cell therapy. Meanwhile, it is often required that NSCs are stored and transported to a long distance for research or treatment purposes. Although high survival rates could be maintained, conventional methods for cell transportation (dry ice or liquid nitrogen) are inconvenient and expensive. Therefore, the establishment of a safe, affordable, and low-cost strategy to store and transport easily accessible hiPSCs and hiNSCs, with characteristics that match fetal hNSCs, is incredibly urgent.
METHODS
We reprogrammed human urinary cells to iPSCs using a non-integrating, virus-free technique and differentiated the iPSCs toward iNSCs/neurospheres and neurons, under Good Manufacturing Practice (GMP)-compatible conditions. The pluripotency of iPSCs and iNSCs was characterized by a series of classical methods (surface markers, karyotype analysis, and in vitro as well as in vivo differentiation capabilities, etc.).
RESULTS
Here, our results showed that we successfully generated hiNSCs/neurospheres from more available, non-invasive, and more acceptable urinary cells by a virus-free technique. Next, we demonstrated that the iNSCs differentiated into mature cerebral cortical neurons and neural networks. Interestingly, hiNSCs survived longer as neurospheres at ambient temperature (AT) than those cultured in a monolayer. Within 7 days approximately, the neural viability remained at > 80%, while hiNSCs cultured in a monolayer died almost immediately. Neurospheres exposed to AT that were placed under standard culture conditions (37 °C, 5% CO) recovered their typical morphology, and retained their proliferation and differentiation abilities.
CONCLUSIONS
In this study, we provided a simple method for the storage of NSCs as neurospheres at AT as an alternative method to more costly and inconvenient traditional methods of cryopreservation. This will enable hiNSCs to be transported over long distances at AT and facilitate the therapeutic application of NSCs as neurospheres without any further treatment.
Topics: Cell Differentiation; Cell- and Tissue-Based Therapy; Cells, Cultured; Humans; Induced Pluripotent Stem Cells; Neural Stem Cells; Neurons
PubMed: 33736654
DOI: 10.1186/s13287-021-02238-4