scholarly journals The effect of storage conditions on samples for the evaluation of copper status in blesbok (Damaliscus pygargus phillipsi)

2002 ◽  
Vol 73 (3) ◽  
Author(s):  
M. Quan ◽  
M.S. Mulders ◽  
D.G.A. Meltzer
Keyword(s):  
Superoxide Dismutase ◽  
Liquid Nitrogen ◽  
Liver Tissue ◽  
Room Temperature ◽  
Storage Conditions ◽  
Superoxide Dismutase Activity ◽  
Activity Levels ◽  
Sample Storage ◽  
Live Animal ◽  
Jersey Cows

Investigaltions to determine the effect of sample storage on the concentration of copper in liver tissue and on the activity of erythrocyte superoxide dismutase were undertaken in preparation for a study of blesbok (Damaliscus pygargus phillipsi) that were suspected to be suffering from copper deficiency. Two liver samples were collected from each of 20 culled blesbok in a manner that simulated the collection of biopsies from the live animal. These samples were stored either in 10 % formalin or frozen at -20 °C until analysed 4 1/2 months later. The effect of different methods of sample storage on superoxide dismutase activity was determined. Erythrocytes collected from 3 Jersey cows and 5 culled blesbok were washed and divided into 0.5m portions, stored at room temperature (~20 °C), in a refrigerator (4 °C), frozen at -20 °C in a freezer, and in liquid nitrogen (-200 °C). An analysis of superoxide dismutase activity was undertaken using a commercial assay kit at intervals of 2-4 days until the levels of activity had fallen significantly. The copper concentration in formalin-preserved liver samples was significantly lower than that measured in frozen liver tissue apparently as a result of leaching. The activity of superoxide dismutase in cattle blood was unchanged for 4 days at room temperature but fell appreciably after 2 days at 4 °C and -20 °C. Enzyme activity remained unchanged for 200 days in erythrocytes stored in liquid nitrogen. Superoxide dismutase activity levels in healthy blesbok were considerably lower than those measured in Jersey cows and remained unaffected for up to 6 days in samples stored at 4 °C and 20 °C. The level of activity fell significantly thereafter. Samples stored in liquid nitrogen were unchanged after 40 days.

A study of superoxide dismutase activity in the liver tissue and the serum of the patients with viral hepatitis.

Kanzo ◽  
1987 ◽  
Vol 28 (6) ◽  
pp. 681-686 ◽  
Author(s):  
Hitoshi TOGASHI ◽  
Haruhide SHINZAWA ◽  
Hiroto WAKABAYASHI ◽  
Nobuo YAMADA ◽  
Touichirou NAKAMURA ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Viral Hepatitis ◽  
Liver Tissue ◽  
Superoxide Dismutase Activity

Superoxide dismutase activity levels in a Spanish population 50-93 years

1999 ◽  
Vol 11 (1) ◽  
pp. 45-47 ◽  
Author(s):  
R. De La Torre ◽  
A. Casado ◽  
M.E. L�pez-Fern�ndez ◽  
D. Carrascosa ◽  
D. Venarucci
Keyword(s):  
Superoxide Dismutase ◽  
Superoxide Dismutase Activity ◽  
Spanish Population ◽  
Activity Levels

scholarly journals Systematic Analysis of Impact of Sampling Regions and Storage Methods on Fecal Gut Microbiome and Metabolome Profiles

mSphere ◽  
2020 ◽  
Vol 5 (1) ◽  
Author(s):  
Yali Liang ◽  
Tianyu Dong ◽  
Minjian Chen ◽  
Lianping He ◽  
Tingzhang Wang ◽  
...  
Keyword(s):  
Gut Microbiome ◽  
Room Temperature ◽  
Storage Conditions ◽  
Minimal Effect ◽  
Sample Storage ◽  
Short Term ◽  
Room Temperature Storage ◽  
Stool Samples ◽  
And Storage ◽  
Temperature Storage

ABSTRACT The contribution of human gastrointestinal (GI) microbiota and metabolites to host health has recently become much clearer. However, many confounding factors can influence the accuracy of gut microbiome and metabolome studies, resulting in inconsistencies in published results. In this study, we systematically investigated the effects of fecal sampling regions and storage and retrieval conditions on gut microbiome and metabolite profiles from three healthy children. Our analysis indicated that compared to homogenized and snap-frozen samples (standard control [SC]), different sampling regions did not affect microbial community alpha diversity, while a total of 22 of 176 identified metabolites varied significantly across different sampling regions. In contrast, storage conditions significantly influenced the microbiome and metabolome. Short-term room temperature storage had a minimal effect on the microbiome and metabolome profiles. Sample storage in RNALater showed a significant level of variation in both microbiome and metabolome profiles, independent of the storage or retrieval conditions. The effect of RNALater on the metabolome was stronger than the effect on the microbiome, and individual variability between study participants outweighed the effect of RNALater on the microbiome. We conclude that homogenizing stool samples was critical for metabolomic analysis but not necessary for microbiome analysis. Short-term room temperature storage had a minimal effect on the microbiome and metabolome profiles and is recommended for short-term fecal sample storage. In addition, our study indicates that the use of RNALater as a storage medium of stool samples for microbial and metabolomic analyses is not recommended. IMPORTANCE The gastrointestinal microbiome and metabolome can provide a new angle to understand the development of health and disease. Stool samples are most frequently used for large-scale cohort studies. Standardized procedures for stool sample handling and storage can be a determining factor for performing microbiome or metabolome studies. In this study, we focused on the effects of stool sampling regions and stool sample storage conditions on variations in the gut microbiome composition and metabolome profile.

scholarly journals Oxidative stress and ageing in Caenorhabditis elegans

1993 ◽  
Vol 292 (2) ◽  
pp. 605-608 ◽  
Author(s):  
J R Vanfleteren
Keyword(s):  
Oxidative Stress ◽  
Superoxide Dismutase ◽  
Caenorhabditis Elegans ◽  
Life Span ◽  
Superoxide Anion ◽  
Superoxide Dismutase Activity ◽  
Activity Levels ◽  
Superoxide Anion Production ◽  
Specific Increase ◽  
The Mean

Mutations in the age-1 gene double both the mean and maximum life span of Caenorhabditis elegans. They also result in an age-specific increase of catalase and Cu/Zn superoxide dismutase activity levels. The higher superoxide dismutase activity levels in age-1 mutants confer hyperresistance to the superoxide-anion-generating drug paraquat. The rate of superoxide anion production by microsome fractions declines linearly with age in age-1(+) worms, but, after an initial decline, is stabilized at a higher level in senescent age-1 mutant nematodes. These results clearly show that oxidative stress resistance and potential life span are correlated in this organism, and they suggest that the natural product of age-1 either directly or indirectly downregulates the activities of several other genes as a function of age.

138 CELL INTEGRITY OF HOLSTEIN AND JERSEY EMBRYOS: COMPARISON OF TWO FREEZING METHODS

2008 ◽  
Vol 20 (1) ◽  
pp. 149
Author(s):  
R. Dupras ◽  
J. Dupras ◽  
Y. Chorfi
Keyword(s):  
Liquid Nitrogen ◽  
Standard Method ◽  
Propidium Iodide ◽  
Room Temperature ◽  
Holstein Cows ◽  
Live Cells ◽  
Cell Integrity ◽  
Hoechst 33342 ◽  
Jersey Cows ◽  
Donor Cows

The aim of this study was to compare cell integrity of Holstein and Jersey embryos using the standard method of freezing and vitrification. Embryos were harvested and processed following standard procedures approved by IETS (International Embryo Transfer Society), and then underwent two methods of freezing. (1) Standard method: embryos were put into 1.5 m ethylene glycol (EG) for 5 min at room temperature, and then at –6.5�C for 10 min for stabilization. After seeding, the temperature was lowered by 0.5�C until it reached –32�C, and then straws containing embryos were plunged into liquid nitrogen. (2) Vitrification: embryos were put for 3 min into vitrification solutions V1: 5 m EG in EmCare (ICPbio, Ltd., Auckland, New Zealand) and V2: 7 m EG, 0.5 m galactose (Sigma Chemical Co., St. Louis, MO, USA), and 18% Ficoll70 (Sigma) in EmCare for 45 s. Embryos were loaded into straws and then plunged into liquid nitrogen. De-freezing was done as follows: 5 s at room temperature, and then 30 s in water at 25�C. Vitrified embryos were de-frozen for 10 s at room temperature followed by 30 s in water at 25�C. All embryos were subjected to a combined staining, Hoechst 33342 and propidium iodide (Sigma-Aldrich Canada, Toronto, Ontario, Canada). The combined staining with these dyes makes analysis of cell death possible. Propidium iodide is specific to dead cells, whereas Hoechst stains all of the cells. Embryos were put in a PBS solution containing 20 µg mL–1 of Hoechst 33342 and propidium iodide for 15 min at 37�C and placed between a slide and coverslip. Fluorescence microscopy was then used to assess blue nuclei (live cells) and red nuclei (dead cells) of the cow embryos. For this experiment, a total of 51 grade 1 embryos (IETS classification) at early blastocyst or blastocyst stages were used (25 Holstein embryos from 4 donor cows and 26 Jersey embryos from 3 donor cows). The standard method of freezing was performed on 26 embryos (13 from Holstein cows and 13 from Jersey cows) and vitrification was performed on 25 embryos (12 from Holstein cows and 13 from Jersey cows). Embryos from a given cow (Holstein or Jersey) were evenly allocated to the standard and vitrification methods. The GLM procedure of SAS (SAS statistical software version 8; SAS Institute, Inc., Cary, NC, USA) was used to compare numbers of dead and live cells between vitrification and the standard method of freezing in Holstein and Jersey embryos. The mean numbers (�SE) of live cells of Holstein embryos were, respectively, 87.92 � 8.48 and 85.5 � 6.46 for standard method and vitrification. For Jersey embryos, means (�SE) of live cells were 94.42 � 2.6 and 73.93 � 7.39, respectively, for the standard method and vitrification. Means (�SE) of dead cells of Holstein embryos were, respectively, 5.58 � 1.37 and 13.67 � 2.91 for the standard method and vitrification. For Jersey embryos, means (�SE) of dead cells were 8.08 � 1.01 and 27.6 � 7.06, respectively, for the standard method and vitrification. In conclusion, vitrification significantly increased (P ≤ 0.05) the number of dead cells of embryos over the standard method of freezing. This effect was more evident (P ≤ 0.05) in Jersey than in Holstein embryos. The authors thank Dr. Vincent Girard for his help with statistics.

scholarly journals Alcohol keeps eDNA at the party longer

2021 ◽  
Vol 4 ◽  
Author(s):  
Sarah Licul ◽  
Rachael Impey ◽  
Andrew Weeks
Keyword(s):  
Room Temperature ◽  
Field Studies ◽  
Dna Degradation ◽  
Storage Conditions ◽  
Sample Storage ◽  
Time Intervals ◽  
Silica Bead ◽  
Scientific Analysis ◽  
Protocol Optimisation ◽  
Water Filters

For a typical eDNA water study, water will be filtered on site, before prompt transfer to a laboratory for DNA extraction and required scientific analysis. In a setting where transport is quick and available, this is a straightforward process. However, many of our studies can occur in remote Australia where sample preservation presents many logistical challenges. Typically, we advise clients to store eDNA water filters after sampling below 4 °C to ensure minimal DNA degradation. For many clients however, field studies often occur in an isolated setting without adequate refrigeration facilities, and as such present challenges for this process. Rather than compromise on sample integrity, EnviroDNA conducted a pilot study into the use of alternate preservation methods on our most commonly used 0.22 mm Sterivex filters. With help from our friendly neighbourhood goldfish tank, our standard 4 °C protocol was compared to a variety of conditions including filled ethanol filters, flushed ethanol filters, lysis buffer and silica bead storage conditions at both 4 °C and room temperature. The study, conducted at various time points over 14-days, used qPCR to quantify the amount of DNA extracted from the filter. Our results revealed that storage within or using flushed ethanol, allowed the samples to be stored for longer time intervals at room temperature, with similar, or in some cases, improved DNA elutions. This protocol optimisation has allowed us to offer an alternate sample storage protocol for clients, expanding the availability and accessibility of eDNA biodiversity assessments around Australia.

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