scholarly journals Analysis of IL-1βRelease from Cryopreserved Pooled Lymphocytes in Response to Lipopolysaccharide and Lipoteichoic Acid

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Sreelekshmi R. Nair ◽  
C. S. Geetha ◽  
P. V. Mohanan

Pyrogens are heterogeneous group of fever-inducing substances derived from Gram-positive and Gram-negative bacteria, fungi, and viruses. They incite immune response by producing endogenous pyrogens such as prostaglandins and other proinflammatory cytokines like IL-1β, IL-6, and TNF-α. The present study was to analyze the influence of cryopreservation in IL-1βrelease, a marker for inflammatory response from human lymphocytes, in response to exogenous pyrogenic stimulants. Lymphocytes isolated from pooled blood of multiple healthy individuals were cryopreserved in DMSO and glycerol for periods of 7, 14, 30, and 60 days and were challenged with LPS and LTAin vitro. The inflammatory cytokine, IL-1βrelease, was measured by ELISA method. It was observed that the release of IL-1βincreases instantaneously after the initiation of incubation and reaches a maximum at 3 to 5 hours and then gradually decreases and gets stabilized for both pyrogens. Moreover it was also observed that the effect of cryoprotectants, DMSO (10%) and glycerol (10%), showed almost similar results for short-term storage, but DMSO-preserved lymphocytes yielded a better viability for long-term storage. Thus, the isolated cryopreserved lymphocytes system can be a promising approach for the total replacement/alteration to animal experimentation for pyrogenicity evaluation.

1998 ◽  
Vol 87 (9) ◽  
pp. 1062-1068 ◽  
Author(s):  
Bruce A. Kerwin ◽  
Martin C. Heller ◽  
Steven H. Levin ◽  
Theodore W. Randolph

2016 ◽  
Vol 9 (3) ◽  
pp. 379-388 ◽  
Author(s):  
N. De Clercq ◽  
G. Vlaemynck ◽  
E. Van Pamel ◽  
D. Colman ◽  
M. Heyndrickx ◽  
...  

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.


2005 ◽  
Vol 6 (2) ◽  
pp. 72-79 ◽  
Author(s):  
Filiz Yalcin

Abstract This study investigated weight changes of seven different light-cured composite restorative materials, one polyacid glass ionomer compomer, and one light-cured glass-ionomer cement following short-term and long-term storage in water. Two packable composites, three universal (hybrid) composites, one microglass composite, one polyacid glass ionomer resin composite (compomer), one microhybrid low-viscosity (flowable) composite, and one light cured glass ionomer composite cement were evaluated in this study. The weight changes of these specimens were measured daily (short-term storage), and they were measured after six weeks (long-term storage) using an electronic analytical balance. A significant difference was found in Ionoliner, Dyract AP, Opticor flow, Charisma, and Solitare 2, but no significant difference was found in the others (Filtek Z 250, Filtek P60, TPH Spectrum, and Valux Plus). Weight change showed a tendency to increase with the time of water storage. The greatest weight change occurred in light-cured glass ionomer composite cement (Ionoliner), which is followed in order by the weight changes in Dyract AP, Opticor Flow, Charisma, Solitare 2, Filtek Z250, Filtek P60, TPH Spectrum; Valux Plus had the least amount of change. Citation Keyf F, Yalcin F. The Weight Change of Various Light-Cured Restorative Materials Stored in Water. J Contemp Dent Pract 2005 May;(6)2:072-079.


2012 ◽  
Vol 37 (1) ◽  
pp. 45-53 ◽  
Author(s):  
EH Mobarak ◽  
LE Daifalla

Clinical Relevance A mild acetone-based single-step self-etched adhesive system may reveal less nanoleakage in the short-term interval, but unfortunately, this was not sustained after long-term storage.


HortScience ◽  
2019 ◽  
Vol 54 (1) ◽  
pp. 143-148 ◽  
Author(s):  
Brianna L. Ewing ◽  
Gregory M. Peck ◽  
Sihui Ma ◽  
Andrew P. Neilson ◽  
Amanda C. Stewart

Hard cider production in the United States has increased dramatically during the past decade, but there is little information on how harvest and postharvest practices affect the chemistry of the resulting cider, including concentrations of organoleptically important flavanols. For 2 years we assessed fruit, juice, and cider from a total of five apple (Malus ×domestica Borkh.) cultivars in two experiments: sequential harvests and postharvest storage. Different cultivars were used in 2015 and 2016 with the exception of ‘Dabinett’, which was assessed in both years. There were no differences in polyphenol concentrations in cider made from fruit that was harvested on three separate occasions over a 4-week period in either 2015 or 2016. Fruit storage durations and temperatures had little influence on the chemistry when the experiment was conducted in 2015, but polyphenol concentration was greater after storage in the 2016 experiment. In 2016, total polyphenols in ‘Dabinett’ ciders were 51% greater after short-term storage at 10 °C and 67% greater after long-term storage at 1 °C than the control, which was not subjected to a storage treatment. In 2016, total polyphenols in ‘Binet Rouge’ ciders were 67% greater after short-term storage at 10 °C and 94% greater after long-term storage at 1 °C than the control. Although results varied among cultivars and harvest years, storing apples for longer periods of time and at warmer temperatures may be a strategy to increase polyphenol, particularly flavanol, concentrations in hard cider.


Plants ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1655
Author(s):  
Soňa Felšöciová ◽  
Przemysław Łukasz Kowalczewski ◽  
Tomáš Krajčovič ◽  
Štefan Dráb ◽  
Miroslava Kačániová

Contamination of malting barley grain and malt with micromycetes sampled at various periods post-harvest (3rd, 6th, and 9th month of storage) and types of storage (storage silo and floor warehouse) was investigated. Each of these barley grain samples was malted. This article reports on the changes in the fungal microbiome composition and their overall count in barley grain and malt. From the surface-disinfected barley grain samples collected immediately after harvest, there were eight genera isolated, with a predominance of Alternaria. A small increase of isolated microfungi was detected in barley stored in silo for 3 and 6 months (from 142 isolates to 149) and decreased below the number of isolates in barley before storage (133 isolates). Fungal count during storage gradually decreased up to 9 month in barley stored in floor warehouse (from 142 isolates to 84). The initial total count of microscopic fungi in malt before storage was the highest (112 isolates) with 7 genera detected, compared to malts prepared from barley stored for longer time (54 isolates, 7 genera, 9th month of storage). Alternaria was the most abundant and frequent genus. Quantitative representation of the filamentous microscopic fungi was lower compared to yeasts especially in barley and malt prepared from barley stored at third month of storage in both type of storage. Yeasts were identified from all grain samples and malt samples with mass spectrometry. Most attention was given to the widely distributed fungus Penicillium, 79% of strains produced at least one mycotoxin detected under in vitro assays using the TLC method (97% of them produced griseofulvin, 94% CPA, 79% patulin, 14% roquefortin C, and penitrem A was produced by two screening strains under laboratory conditions). It is therefore important to monitor the microflora throughout the production cycle of “barley to beer”.


2021 ◽  
Vol 8 ◽  
Author(s):  
Giscard Lima ◽  
Alexander Kolliari-Turner ◽  
Fernanda Rossell Malinsky ◽  
Fergus M. Guppy ◽  
Renan Paulo Martin ◽  
...  

Introduction: Recombinant human erythropoietin (rHuEPO) administration studies involving transcriptomic approaches have demonstrated a gene expression signature that could aid blood doping detection. However, current anti-doping testing does not involve collecting whole blood into tubes with RNA preservative. This study investigated if whole blood in long-term storage and whole blood left over from standard hematological testing in short-term storage could be used for transcriptomic analysis despite lacking RNA preservation.Methods: Whole blood samples were collected from twelve and fourteen healthy nonathletic males, for long-term and short-term storage experiments. Long-term storage involved whole blood collected into Tempus™ tubes and K2EDTA tubes and subjected to long-term (i.e., ‒80°C) storage and RNA extracted. Short-term storage involved whole blood collected into K2EDTA tubes and stored at 4°C for 6‒48 h and then incubated at room temperature for 1 and 2 h prior to addition of RNA preservative. RNA quantity, purity, and integrity were analyzed in addition to RNA-Seq using the MGI DNBSEQ-G400 on RNA from both the short- and long-term storage studies. Genes presenting a fold change (FC) of >1.1 or < ‒1.1 with p ≤ 0.05 for each comparison were considered differentially expressed. Microarray analysis using the Affymetrix GeneChip® Human Transcriptome 2.0 Array was additionally conducted on RNA from the short-term study with a false discovery ratio (FDR) of ≤0.05 and an FC of >1.1 or < ‒1.1 applied to identify differentially expressed genes.Results: RNA quantity, purity, and integrity from whole blood subjected to short- and long-term storage were sufficient for gene expression analysis. Long-term storage: when comparing blood tubes with and without RNA preservation 4,058 transcripts (6% of coding and non-coding transcripts) were differentially expressed using microarray and 658 genes (3.4% of mapped genes) were differentially expressed using RNA-Seq. Short-term storage: mean RNA integrity and yield were not significantly different at any of the time points. RNA-Seq analysis revealed a very small number of differentially expressed genes (70 or 1.37% of mapped genes) when comparing samples stored between 6 and 48 h without RNA preservative. None of the genes previously identified in rHuEPO administration studies were differently expressed in either long- or short-term storage experiments.Conclusion: RNA quantity, purity, and integrity were not significantly compromised from short- or long-term storage in blood storage tubes lacking RNA stabilization, indicating that transcriptomic analysis could be conducted using anti-doping samples collected or biobanked without RNA preservation.


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