Effect of storage temperature on spore viability and early gametophyte development of three vulnerable species of Alsophila (Cyatheaceae)

2010 ◽  
Vol 58 (2) ◽  
pp. 89 ◽  
Author(s):  
Y. Li ◽  
Y. L. Zhang ◽  
C. D. Jiang ◽  
T. Wang ◽  
Q. Wang ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Room Temperature ◽  
Storage Temperature ◽  
Vulnerable Species ◽  
Spore Viability ◽  
Great Loss ◽  
Gametophyte Development ◽  
Long Term Storage ◽  
Duration Of Storage

To effectively preserve the vulnerable species of Alsophila, we studied the effects of varying the temperature and duration of storage on spore viability, early gametophyte development and the microstructure of brown spores of three Alsophila species. Spores of A. spinulosa (Wall. ex Hook.) Tryon and A. gigantea Wall. ex Hook. lost viability quickly when stored at room temperature and suffered from great loss when stored at –18°C from 6 to 12 months. Within 1 month, spore viability of A. spinulosa and A. gigantea stored at 4°C was higher than that of those stored in liquid nitrogen. In contrast, long-term storage in liquid nitrogen resulted in a comparatively small loss of viability for these two species. The spores of A. podophylla Hook. died within 3 months after storage at room temperature, 4°C and –18°C, and they died within 12 months when stored in liquid nitrogen. The spores of A. spinulosa and A. gigantea stored at room temperature, 4°C and –18°C, were prone to develop into abnormal gametophytes. These results suggest that storage of A. spinulosa and A. gigantea spores in liquid nitrogen is an effective method of preserving these vulnerable species. The reasons for the failure to preserve ephemeral A. podophylla spores by storage in liquid nitrogen are discussed.

scholarly journals Development of cryopreservation methods of seed of Nepeta cataria

2021 ◽  
Vol 102 (2) ◽  
pp. 37-42
Author(s):  
Margarita Ishmuratova ◽  
◽  
Damirzhan Baigarayev ◽  
Saltanat Tleukenova ◽  
Elena Gavrilkova ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Room Temperature ◽  
Seed Viability ◽  
Positive Temperature ◽  
Seed Rate ◽  
Nepeta Cataria ◽  
Long Term Storage ◽  
Medical Plant ◽  
Term Storage

This article presents the summarized data on cryopreservation of seeds of the medical plant Nepeta cataria. Cryopreservation is a highly promising method for saving of seed materials, allowing to organize long-term storage without viability loss. The purpose of present work is to optimize conditions of cryopreservation of seed materials of Nepeta cataria. Assessment of seed survival rate in the storage showed a linear decrease in seed viability and energy of germination. After 30 months of storage at the low positive temperature (+5 ºC) in paper pack seed rate decreased to 12.0 % and energy of germination to 11.2 %; after 4 years of storage seeds lost viability. During conduction of research the type of container, condition of thawing, optimal moisture of seeds and cryoprotectants are optimized. The optimal container for cryopreservation in liquid nitrogen was plastic cryo tubes; defrosting at room temperature. The best seed rate is found at moisture 3 %; the best cryoprotectant was glucose, the optimal concentration was 15 %. The result of the research is used for creation of the long-term storage medicinal cultures’ seed bank in the liquid nitrogen.

scholarly journals Spore viability of microsporidian species isolated from gipsy moth larvae (Lymantria dispar) after long-term storage in liquid nitrogen

2019 ◽  
Vol 143 (7-8) ◽  
pp. 323-324
Author(s):  
Daniela Pilarska ◽  
Andreas Linde ◽  
Gernot Hoch ◽  
Manana Kereselidze
Keyword(s):  
Liquid Nitrogen ◽  
Lymantria Dispar ◽  
Spore Viability ◽  
Microsporidian Species ◽  
Long Term Storage ◽  
Gipsy Moth ◽  
Term Storage

U radu se prikazuju rezultati preživljenja mikrosporidija izoliranih iz gubara (Lymantria dispar) nakon dugotrajne pohrane u tekućem dušiku. Infektivnost osam mikrosporidijskih L. dispar izolata testirano je na ličinkama gubara: Vairimorpha disparis, Nosema lymantriae, Nosema portugal, Nosema sp. (Poljska), Nosema sp. (Ebergassing), Nosema sp. (Njemačka), Nosema sp. (Schweinfurt) and Nosema sp. (Veslec). Preživljenje spora u tekućem dušiku detaljno je praćeno kod N. portugal i Nosema sp. (Ebergassing) koje su tako čuvane skoro 19 godina i aplicirane oralnom infekcijom i ponaosob na svaku pojedinu ličinku gubara. Ostalih 6 izolata aplicirano je površinskom kontaminacijom hranjivog supstrata, također oralnim infekcijskim putem. Od 8 mikrosporidijskih izolata apliciranih površinskom kontaminacijom samo kod 4 izolata, Vairimorpha disparis, Nosema lymantriae, Nosema sp. (Ebergassing) i Nosema sp. (Poljska), došlo je do uspješne infekcije ličinki gubara. Dok je kod svih testiranih ličinki (100%) uspješno realizirana infekcija sporama prve tri mikrosporidije, tek 21,1% tretiranih ličinki uspješno je inficirano sporama Nosema sp. (Poljska). Na kraju inkubacijskog razdoblja, kod svih ličinki inficiranih sa sva 4 izolata disekcijom je utvrđen visok stupanj zaraze; tkiva su bila ispunjena sporama. Mikroskopska pretraga ličinki L. dispar individualno tretiranih sporama Nosema portugal uz tri različite doze (100, 1000 i 10000 spora) pokazala je da su spore N. portugal izgubile infektivnost nakon 19 godina pohrane u tekućem dušiku; niti jedna od testiranih ličinka nije bila zaražena. Naprotiv, spore Nosema sp. (Ebergassing) zadržale su infektivnost nakon istog razdoblja pohrane u tekućem dušiku od 18,75 godina. Niti jedna od testiranih ličinki nije zaražena nakon oralnog unosa od 100 spora. Oralna aplikacija od 1000 spora po ličniki rezultirala je ukupnom uspješnom infekcijom 4,1% ličinki, a aplikacija od 10000 spora s 68,8% inficiranih ličinki. U svim uspješnim slučajevima uspješno zaraženih ličinki, na kraju inkubacijskog razdoblja infekcija je bila dobro razvijena, a masno tijelo prepuno spora. Istraživanje je potvrdilo da je pohrana mikrosporidija Nosema i Vairimorpha vrsta i domaćina iz reda leptira prikladna opcija za dugotrajno čuvanje izolata. Spore su preživjele i do 18 i pola godina, iako je tijekom eksperimenta zamijećen i značajan pad njihove infektivnosti. U pojedinim slučajevima spore su odumrle već nakon 7 godina pohrane u tekućem dušiku. Preporuka je da prilikom skladištenja i pohrane mikrosporidijskih izolata u tekućem dušiku svakih 5 godina repozitorij obnavlja svježim izolatima. Materijal koji se dulje vrijeme skladišti u tekućem dušiku nije prikladan i ne bi se smio koristiti u infektološkim testovima.

Efficacy of the Antigenicity-Retaining Ability of Fixative Solutions for Liquid-Based Cytology: Immunocytochemistry of Long-Term Storage

2021 ◽  
pp. 1-12
Author(s):  
Hiaki Sato ◽  
Yoshiaki Norimatsu ◽  
Satoshi Irino ◽  
Takeshi Nishikawa
Keyword(s):  
Cell Membrane ◽  
Room Temperature ◽  
Storage Temperature ◽  
Storage Period ◽  
Storage Conditions ◽  
Immunocytochemical Staining ◽  
Long Term Storage ◽  
Liquid Based Cytology ◽  
Term Storage

<b><i>Introduction/Objective:</i></b> Liquid-based cytology (LBC) is advantageous as multiple stained specimens can be prepared and used for additional assays such as immunocytochemical and molecular-pathological investigations. Two types of preservative-fixative solutions (fixatives) are used for nongynecologic specimens used in the BD SurePath-LBC (SP-LBC) method, and their components vary. However, few studies have evaluated the differences in antigen-retaining ability between these fixatives. Therefore, we investigated and compared the antigen-retaining ability of the fixatives in immunocytochemical staining (ICC) under long-term storage conditions. <b><i>Materials and Methods:</i></b> Sediments of cultured RAJI cells (derived from Burkitt’s lymphoma) were added to each fixative (red and blue) and stored at room temperature for a specified period (1 h; 1 week; and 1, 3, and 6 months). The specimens were then prepared using the SP-LBC method and subjected to ICC. Positivity rate was calculated using the specimens fixed at room temperature for 1 h as a control. Antibodies against Ki67 expressed in the nucleus and against CD20 and leukocyte common antigen (LCA) expressed on the cell membrane were used. <b><i>Results:</i></b> For CD20 and LCA, the positivity rate increased with time in the red fixative compared with that in the control. In the blue fixative, the positivity rate was highest at 1 h and was maintained at a high level throughout the storage period. In contrast, the Ki67 positivity rate was highest at 1 h in both red and blue fixatives and markedly decreased with time. Therefore, although refrigerated (8°C) storage was used, no improvement was noted. <b><i>Conclusions:</i></b> Long-term storage is possible for cell membrane antigens at room temperature; however, it is unsuitable for intranuclear antigens. Therefore, we conclude that suitable fixative type and storage temperature differ based on antigen location. Further investigation is warranted.

The Effect of Storage Temperature and Physical State on the Performance of Antisera in Radioimmunoassay after Long-Term Storage

1988 ◽  
Vol 25 (1) ◽  
pp. 89-95
Author(s):  
B A Middleton ◽  
L M Morgan ◽  
G W Aherne ◽  
V Marks
Keyword(s):  
Room Temperature ◽  
Storage Temperature ◽  
Storage Conditions ◽  
Liquid Form ◽  
Long Term Storage ◽  
Freeze Dried ◽  
Control Serum ◽  
Term Storage ◽  
Temperature Storage

The performance in radioimmunoassay of four antisera after storage at temperatures ranging from −40°C to room temperature, in three physical states (frozen, liquid or freeze dried) was investigated over a 3-year period. No deterioration in antiserum performance in terms of precision and accuracy of quality control serum measurement or recovery of ligand was apparent under any of the storage conditions studied. Some lowering of titre became apparent in two of the antisera over the study period. Deterioration was most marked when antiserum was stored lyophilised at room temperature. Storage of antiserum frozen confers no advantage over storage at 4°C provided precautions are taken to minimise bacterial contamination when storing antiserum in liquid form.

scholarly journals Cryopreservation of Serbian autochthonous plum ‘Crvena Ranka’ using aluminium cryo-plates

Genetika ◽  
2021 ◽  
Vol 53 (1) ◽  
pp. 283-294
Author(s):  
Tatjana Vujovic ◽  
Darko Jevremovic ◽  
Tatjana Marjanovic ◽  
Djurdjina Ruzic
Keyword(s):  
Ethylene Glycol ◽  
Liquid Nitrogen ◽  
Room Temperature ◽  
Genetic Erosion ◽  
Ms Medium ◽  
Plate Method ◽  
Long Term Storage ◽  
Commercially Important ◽  
Term Storage

?Crvena Ranka? is one of the oldest autochthonous plum cultivars in Serbia. However, it is endangered due to the long-term pathogen pressure and continuous suppression by commercially important cultivars and therefore faced with genetic erosion. This study was carried out to investigate the suitability of two efficient and simple cryopreservation methods using aluminium cryo-plates for its conservation. Precultured shoot tips of this genotype were embedded in alginate gel into cryo-plates wells. Osmoprotection was performed using loading solution with 1.9 M glycerol and 0.5 M sucrose (30 min at room temperature). In the V cryo-plate protocol, explants were dehydrated at room temperature for 20 or 40 min with PVS A3 (37.5% glycerol, 15% dimethylsulfoxide, 15% ethylene glycol and 22.5% sucrose) or with PVS3 (50% glycerol and 50% sucrose) for 60 min. In the D cryo-plate protocol, desiccation for 2, 2.5 or 3 h was performed over silica gel. Then, the cryo-plates were directly immersed into liquid nitrogen. Unloading was done in MS medium containing 0.8 M sucrose (30 min at room temperature). In the V cryo-plate procedure regrowth of cryopreserved explants dehydrated with PVS A3 was between 50% and 51.9%, while in those dehydrated with PVS3 it was 66.7%. As for the D cryo-plate method, regrowth of cryopreserved explants ranged between 30-40%. After regrowth, shoots were successfully multiplied and rooted. Results prove the feasibility of these new cryogenic methods for a long-term storage of this valuable Prunus genotype.

Evaluation of long-term frozen storage of plasma for measurement of high-density lipoprotein and its subfractions by precipitation

1990 ◽  
Vol 36 (5) ◽  
pp. 783-788 ◽  
Author(s):  
M N Nanjee ◽  
N E Miller
Keyword(s):  
High Density Lipoprotein ◽  
Liquid Nitrogen ◽  
Frozen Storage ◽  
Density Lipoprotein ◽  
High Density ◽  
Long Term Stability ◽  
Long Term Storage ◽  
Different Temperatures ◽  
Edta Plasma

Abstract The concentration of high-density lipoprotein cholesterol (HDL-C) in plasma is now established as an independent risk factor for coronary heart disease, but more data are needed on the relative risk-predictive powers of different HDL subclasses. For epidemiologic and clinical purposes, isolation of HDL from other lipoproteins and separation of its two major subclasses, HDL2 and HDL3, are performed most conveniently by precipitation. Although storage of plasma is commonly necessary, little information is available on the long-term stability of HDL subclasses at different temperatures. Therefore, we quantified HDL-C, HDL2-C, and HDL3-C by dual precipitation with heparin-MnCl2/15-kDa dextran sulfate (H-M/DS) in samples of EDTA-plasma from 93 healthy subjects, after storage for one to 433 days at -20 degrees C, at -70 degrees C, or in liquid nitrogen (-196 degrees C). Fourteen samples (15%) were stored for a year or longer. At -20 degrees C, HDL-C decreased by 4.8% per year and HDL3-C decreased by 6.9% per year (P = 0.002 for both variables) relative to results obtained with samples stored in liquid nitrogen; total cholesterol, HDL2-C, and triglyceride did not change significantly at this temperature. When stored at -70 degrees C, none of the lipids showed any change relative to results obtained with liquid nitrogen. Thus, long-term storage of EDTA-plasma at -20 degrees C is unsuitable for subsequent quantification of HDL-C and its subclasses by H-M/DS dual precipitation. Storage at -70 degrees C is preferable, and is as reliable as storage in liquid nitrogen.

NANOEMULSIONS BASED ON LIPOIC ACID DERIVATIVES WITH VARIOUS ALCOHOLS: PRODUCTION AND INFLUENCE ON FUNCTIONAL ACTIVITY OF PLATELETS

2020 ◽  
pp. 461-464
Author(s):  
A. Sinebryukhova ◽  
A. Shipelova ◽  
E. Darnotuk ◽  
A. Chekanov ◽  
O. Baranova ◽  
...  
Keyword(s):  
Particle Size ◽  
Arachidonic Acid ◽  
Lipoic Acid ◽  
Encapsulation Efficiency ◽  
Room Temperature ◽  
Optimal Conditions ◽  
Pluronic F68 ◽  
Long Term Storage ◽  
Term Storage

The optimal conditions were selected for obtaining homogeneous nanoemulsions (NE) of lipoic acid conjugates (LA-conjugates) based on Pluronic F68 (1,8%) with a particle size not exceeding 400 nm, characterized by 97±2% encapsulation efficiency of substances in nanoparticles (NP). A heterogeneous NE (polydispersity index, PDI>0,3) with the derivative of LA and myo-inositol based on phosphatidylcholine (PC, C = 3 mg/ml) was also obtained consisting of 2 particle fractions: 20–70 nm (27%) and 122–212 nm (73%). The obtained NEs with LA-conjugates based on Pluronic F68 and PC were stable during long-term storage (more than 12 months) at room temperature. The effect of the obtained NEs of LA-conjugates on platelet aggregation (Pt) caused by arachidonic acid (AA) was determined, and a mechanism of their action was proposed.

scholarly journals HMGB1 concentration measurements in trauma patients: assessment of pre-analytical conditions and sample material

2019 ◽  
Vol 26 (1) ◽  
Author(s):  
William Ottestad ◽  
Ingrid N. Rognes ◽  
Erlend Skaga ◽  
Cassandra Frisvoll ◽  
Guttorm Haraldsen ◽  
...  
Keyword(s):  
Western Blot ◽  
Room Temperature ◽  
Trauma Patients ◽  
Plasma Samples ◽  
Sample Material ◽  
Limits Of Agreement ◽  
Freeze Thaw ◽  
Long Term Storage ◽  
Term Storage

Abstract Background HMGB1 is a mediator of systemic inflammation in sepsis and trauma, and a promising biomarker in many diseases. There is currently no standard operating procedure for pre-analytical handling of HMGB1 samples, despite that pre-analytical conditions account for a substantial part of the overall error rate in laboratory testing. We hypothesized that the considerable variations in reported HMGB1 concentrations and kinetics in trauma patients could be partly explained by differences in pre-analytical conditions and choice of sample material. Methods Trauma patients (n = 21) admitted to a Norwegian Level I trauma center were prospectively included. Blood was drawn in K2EDTA coated tubes and serum tubes. The effects of delayed centrifugation were evaluated in samples stored at room temperature for 15 min, 3, 6, 12, and 24 h respectively. Plasma samples subjected to long-term storage in − 80 °C and to repeated freeze/thaw cycles were compared with previously analyzed samples. HMGB1 concentrations in simultaneously acquired arterial and venous samples were also compared. HMGB1 was assessed by standard ELISA technique, additionally we investigated the suitability of western blot in both serum and plasma samples. Results Arterial HMGB1 concentrations were consistently lower than venous concentrations in simultaneously obtained samples (arterial = 0.60 x venous; 95% CI 0.30–0.90). Concentrations in plasma and serum showed a strong linear correlation, however wide limits of agreement. Storage of blood samples at room temperature prior to centrifugation resulted in an exponential increase in plasma concentrations after ≈6 h. HMGB1 concentrations were fairly stable in centrifuged plasma samples subjected to long-term storage and freeze/thaw cycles. We were not able to detect HMGB1 in either serum or plasma from our trauma patients using western blotting. Conclusions Arterial and venous HMGB1 concentrations cannot be directly compared, and concentration values in plasma and serum must be compared with caution due to wide limits of agreement. Although HMGB1 levels in clinical samples from trauma patients are fairly stable, strict adherence to a pre-analytical protocol is advisable in order to protect sample integrity. Surprisingly, we were unable to detect HMGB1 utilizing standard western blot analysis.

Patulin production by Penicillium expansum isolates from apples during different steps of long-term storage

2016 ◽  
Vol 9 (3) ◽  
pp. 379-388 ◽  
Author(s):  
N. De Clercq ◽  
G. Vlaemynck ◽  
E. Van Pamel ◽  
D. Colman ◽  
M. Heyndrickx ◽  
...  
Keyword(s):  
Phenotypic Diversity ◽  
Penicillium Expansum ◽  
Penicillium Species ◽  
Cold Temperatures ◽  
Long Term Storage ◽  
In Vitro Investigation ◽  
Duration Of Storage ◽  
Term Storage

Penicillium expansum is the principal cause of blue mould rot and associated production of patulin, a weak mycotoxin, in apples worldwide. P. expansum growth and patulin production is observed during improper or long-term storage of apples. We have investigated the extent to which each successive step during long-term storage contributes to patulin production in various P. expansum isolates. Fungal isolates collected on apples from several Belgian orchards/industries were identified to species level. Random amplification of polymorphic DNA (RAPD) analysis and β-tubulin gene sequencing identified P. expansum and Penicillium solitum as the most prevalent Penicillium species associated with Belgian apples. All 27 P. expansum isolates and eight reference strains were characterised for their patulin production capacity on apple puree agar medium for five days under classical constant temperature and atmosphere conditions. Under these conditions, a large range of patulin production levels was observed. Based on this phenotypic diversity, five P. expansum isolates and one reference strain were selected for in vitro investigation of patulin production under representative conditions in each step of long-term apple storage. Patulin accumulation seemed highly strain dependent and no significant differences between the storage steps were observed. The results also indicated that a high spore inoculum may lead to a strong patulin accumulation even at cold temperatures (1 °C) combined with controlled atmosphere (CA) (3% O2, 1% CO2), suggesting that future control strategies may benefit from considering the duration of storage under CA conditions as well as duration of deck storage.

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