COMPARISON OF LONG-TERM STORAGE IN CHEMICAL FIXATIVES ON MORPHOLOGY AND ANATOMY OF SUPERDWARF WHEAT

Wheat (Triticum aestivum L. cv. SuperDwarf) plants grown in the microgravity of space aboard the Russian space station Mir and sampled for morphological and anatomical comparison to those ex-posed to gravity on earth will be stored in chemical fixative for relatively long periods of time. Our objective was to examine, evaluate, and verify that the integrity of the vascular system and cellular components of wheat plants is maintained following storage in a chemical fixative. In ground-based studies, wheat seedlings were harvested on days 9, 28, and 68 and preserved in 4% formaldehyde: 1% glutaraldehyde (4F: 1G) fixative, adjusted to pH 7.2. Samples of leaves and/or stems were taken from these stocks after various times in storage (24 h, 1, 6, 12, and 24 months), dehydrated, and embedded in Spurr's resin. Semi-thin (1 μm) and thin (50–70 nm) sections were examined by light and transmission electron microscopy. Overall, there seems to be no major artifacts in the cellular structure. The plasmalemma and other organelles appeared normal in this fixative. Use of differential chromophores suggests that enzyme localizations at both the light and electron microscopical levels are not adversely affected by long-term storage. (Supported by NASA Grant NCC 2-831 and the Utah Agr. Expt. Station.)

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Dissolution Characteristics and Morphology of Large-sized Scorodite Particles Synthesized from Fe(II) and As(V) in Aqueous Solution

High Temperature Materials and Processes ◽

10.1515/htmp-2012-0080 ◽

2012 ◽

Vol 31 (4-5) ◽

pp. 451-458 ◽

Cited By ~ 3

Author(s):

S. Fujieda ◽

K. Shinoda ◽

T. Inanaga ◽

M. Abumiya ◽

S. Suzuki

Keyword(s):

Electron Microscopy ◽

Aqueous Solution ◽

Surface Defects ◽

Atmospheric Temperature ◽

Orthorhombic Structure ◽

X Ray ◽

Long Term Storage ◽

Transmission Electron ◽

Term Storage

AbstractA novel process for preparing scorodite particles with a diameter of approximately 20 µm from Fe(II) and As(V) in aqueous solution has been developed by DOWA Metals and Mining. In the present study, the dissolution characteristics of iron and arsenic from the scorodite particles synthesized by this process have been investigated under different conditions. The results show that the concentration of arsenic dissolved from the particles in aqueous solution is very low, but it has a complicated dependence on the temperature and pH of the solution. Transmission electron microscopy (TEM) with an energy dispersive X-ray spectrometer (EDS) was used to analyze the morphology, structure, and composition of the scorodite particles. The results indicate that the scorodite particles exhibit a nearly octahedral shape with planes composed of almost (111) planes in the orthorhombic structure. The concentration of iron at the surface of the particles is higher than that of iron inside of the particles. This characteristic morphology, along with the minimal surface defects of the scorodite particles, is considered to be responsible for the low dissolution of arsenic from the particles in aqueous solution. Atmospheric temperature and solution conditions were also found to be important for the safe, long-term storage of arsenic using scorodite particles.

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A Tissue Retrieval and Postharvest Processing Regimen for Rodent Reproductive Tissues Compatible with Long-Term Storage on the International Space Station and Postflight Biospecimen Sharing Program

BioMed Research International ◽

10.1155/2015/475935 ◽

2015 ◽

Vol 2015 ◽

pp. 1-12 ◽

Cited By ~ 3

Author(s):

Vijayalaxmi Gupta ◽

Lesya Holets-Bondar ◽

Katherine F. Roby ◽

George Enders ◽

Joseph S. Tash

Keyword(s):

Posttranslational Modifications ◽

Space Station ◽

Tissue Storage ◽

High Quality ◽

International Space ◽

Reproductive Tissues ◽

Long Term Storage ◽

Species Specific ◽

Term Storage

Collection and processing of tissues to preserve space flight effects from animals after return to Earth is challenging. Specimens must be harvested with minimal time after landing to minimize postflight readaptation alterations in protein expression/translation, posttranslational modifications, and expression, as well as changes in gene expression and tissue histological degradation after euthanasia. We report the development of a widely applicable strategy for determining the window of optimal species-specific and tissue-specific posteuthanasia harvest that can be utilized to integrate into multi-investigator Biospecimen Sharing Programs. We also determined methods for ISS-compatible long-term tissue storage (10 months at −80°C) that yield recovery of high quality mRNA and protein for western analysis after sample return. Our focus was reproductive tissues. The time following euthanasia where tissues could be collected and histological integrity was maintained varied with tissue and species ranging between 1 and 3 hours. RNA quality was preserved in key reproductive tissues fixed in RNAlaterup to 40 min after euthanasia. Postfixation processing was also standardized for safe shipment back to our laboratory. Our strategy can be adapted for other tissues under NASA’s Biospecimen Sharing Program or similar multi-investigator tissue sharing opportunities.

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Lipophagy contributes to long-term storage of spermatozoa in the epididymis of the Chinese soft-shelled turtle Pelodiscus sinensis

Reproduction Fertility and Development ◽

10.1071/rd18307 ◽

2019 ◽

Vol 31 (4) ◽

pp. 774 ◽

Cited By ~ 5

Author(s):

Hong Chen ◽

Yufei Huang ◽

Ping Yang ◽

Tengfei Liu ◽

Nisar Ahmed ◽

...

Keyword(s):

Molecular Mechanisms ◽

Sperm Storage ◽

Pelodiscus Sinensis ◽

Long Term Storage ◽

Transmission Electron ◽

Epididymal Spermatozoa ◽

Oil Red O Staining ◽

Term Storage ◽

Oil Red O

Spermatozoa are known to be stored in the epididymis of the Chinese soft-shelled turtle Pelodiscus sinensis for long periods after spermiation from the testes, but the molecular mechanisms underlying this storage are largely unknown. In this study, epididymal spermatozoa were investigated to determine the potential molecular mechanism for long-term sperm storage in P. sinensis. Transmission electron microscopy (TEM) and Oil red O staining indicated that unusually large cytoplasmic droplets containing lipid droplets (LDs) were attached to the epididymal spermatozoa. However, the content of LDs decreased gradually with the sperm storage. LDs were surrounded by autophagic vesicles and sequestered as degradative cargo within autophagosome. Immunofluorescence and western blotting demonstrated that autophagy in spermatozoa increased gradually with the storage time. Invitro studies found that spermatozoa obtained from soft-shelled turtles in January can survive more than 40 days at 4°C. Furthermore, immunofluorescence and TEM showed that autophagy was involved in the degradation of LDs with the extension of sperm incubation. Inhibition of autophagy with 3-methyladenine significantly suppressed LD degradation. Moreover, adipose triglyceride lipase was involved in the metabolism of LDs. These findings indicate that lipophagy was activated to maximise LD breakdown, which contributes to long-term sperm storage in the epididymis of P. sinensis.

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Comparison of methods for pre-processing, exosome isolation, and RNA extraction in unpasteurized bovine and human milk

PLoS ONE ◽

10.1371/journal.pone.0257633 ◽

2021 ◽

Vol 16 (9) ◽

pp. e0257633

Author(s):

Sanoji Wijenayake ◽

Shafinaz Eisha ◽

Zoya Tawhidi ◽

Michael A. Pitino ◽

Michael A. Steele ◽

...

Keyword(s):

Human Milk ◽

Biological Fluid ◽

Rna Extraction ◽

Bovine Milk ◽

Protein Markers ◽

Long Term Storage ◽

Transmission Electron ◽

Cellular Debris ◽

Term Storage

Milk is a highly complex, heterogeneous biological fluid that contains non-nutritive, bioactive extracellular vesicles called exosomes. Characterization of milk-derived exosomes (MDEs) is challenging due to the lack of standardized methods that are currently being used for milk pre-processing, storage, and exosome isolation. In this study, we tested: 1) three pre-processing methods to remove cream, fat, cellular debris, and casein proteins from bovine milk to determine whether pre-processing of whole milk prior to long-term storage improves MDE isolations, 2) the suitability of two standard exosome isolation methods for MDE fractionation, and 3) four extraction protocols for obtaining high quality RNA from bovine and human MDEs. MDEs were characterized via Transmission Electron Microscopy (TEM), Nanoparticle Tracking Analysis (NTA), and western immunoblotting for CD9, CD63, and Calnexin protein markers. We also present an optimized method of TEM sample preparation for MDEs. Our results indicate that: 1) Removal of cream and fat globules from unpasteurized bovine milk, prior to long-term storage, improves the MDE yield but not purity, 2) Differential ultracentrifugation (DUC) combined with serial filtration is better suited for bovine MDE isolation compared to ExoQuick (EQ) combined with serial filtration, however both methods were comparable for human milk, and 3) TRIzol LS is better suited for RNA extraction from bovine MDEs isolated by EQ and DUC methods. 4) TRIzol LS, TRIzol+RNA Clean and Concentrator, and TRIzol LS+RNA Clean and Concentrator methods can be used for RNA extractions from human MDEs isolated by EQ, yet the TRIzol LS method is better suited for human MDEs isolated by DUC. The QIAzol + miRNeasy Mini Kit produced the lowest RNA yield for bovine and human MDEs.

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Modelling of Moisture Movement in Wood during Outdoor Storage

Nonlinear Analysis Modelling and Control ◽

10.15388/na.2001.6.1.15210 ◽

2001 ◽

Vol 6 (2) ◽

pp. 3-14 ◽

Cited By ~ 23

Author(s):

R. Baronas ◽

F. Ivanauskas ◽

I. Juodeikienė ◽

A. Kajalavičius

Keyword(s):

Experimental Data ◽

Finite Difference ◽

Climatic Conditions ◽

Two Dimensional ◽

Moisture Movement ◽

Finite Difference Technique ◽

Long Term Storage ◽

Space Formulation ◽

Term Storage

A model of moisture movement in wood is presented in this paper in a two-dimensional-in-space formulation. The finite-difference technique has been used in order to obtain the solution of the problem. The model was applied to predict the moisture content in sawn boards from pine during long term storage under outdoor climatic conditions. The satisfactory agreement between the numerical solution and experimental data was obtained.

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