scholarly journals Study of the preanalytical variables affecting the measurement of clinically relevant free-circulating microRNAs: focus on sample matrix, platelet depletion, and storage conditions

2020 ◽  
Vol 30 (1) ◽  
pp. 83-95 ◽  
Author(s):  
Martina Faraldi ◽  
Veronica Sansoni ◽  
Giovanni Lombardi ◽  
Giuseppe Banfi ◽  
Ewa Ziemann ◽  
...  
Keyword(s):  
Room Temperature ◽  
Quantitative Polymerase Chain Reaction ◽  
Venous Blood ◽  
Storage Conditions ◽  
Circulating Micrornas ◽  
Sample Matrix ◽  
Polymerase Chain ◽  
And Storage ◽  
Plasma Mirnas ◽  
Platelet Depletion

Introduction: Circulating microRNAs (miRNAs) are emerging as potential biomarkers. However, the lack of preanalytical and analytical standardization limits their use. The aim of this study was to determine the expression of different miRNAs in plasma according to different collection and storage conditions. Materials and methods: Venous blood from 10 volunteers was collected in tubes spray-coated with dipotassium salt of ethylendiaminetetraacetic acid, either with (plasma-preparation tube, PPT) or without (K2EDTA) gel separator. Platelet-poor plasma (PPP) was also obtained from K2EDTA plasma. After storage under different conditions, miRNA-enriched total RNA was isolated from plasma and reverse transcribed. A panel of 179 miRNAs was assayed by quantitative polymerase chain reaction and the results were analysed by GenEx software. Detectability and stability of miRNAs were determined. Results: The number of undetected miRNAs was: 18, 24, and 22 in PPT; 83, 43, and 20 in K2EDTA; and 76, 106, and 104 in PPP samples, for plasma immediately frozen at - 80°C and plasma stored for 24h at room temperature or 4°C, respectively. Circulating miRNA expression in PPT samples was not affected by storage delay or temperature, while the percentage of up- and down-regulated miRNA in K2EDTA and PPP samples ranged from 2%, and 1% to 7%, and 5%, respectively. Conclusions: Sample matrix, temperature and delay in storage strongly influence the expression level of plasma miRNAs. Our results indicate PPT tubes as the most suitable matrix to improve total miRNA detectability and stability, independently of temperature.

scholarly journals Dynamic Changes in Plasma MicroRNAs Have Potential Predictive Values in Monitoring Recurrence and Metastasis of Nasopharyngeal Carcinoma

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Xia Xu ◽  
Juan Lu ◽  
Fan Wang ◽  
Xiong Liu ◽  
Xiaohong Peng ◽  
...  
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Treatment Initiation ◽  
Quantitative Polymerase Chain Reaction ◽  
Dynamic Changes ◽  
Predictive Values ◽  
Circulating Micrornas ◽  
Before And After ◽  
Polymerase Chain ◽  
Plasma Mirnas

Although circulating microRNAs (miRNAs) have already proven to be useful as diagnostic and prognostic biomarkers in nasopharyngeal carcinoma (NPC), the potential of these molecules to monitor patients over time has been less explored. This study aimed to analyze dynamic changes in plasma miRNAs before and after treatment and explore their clinical significance in monitoring recurrence and metastasis of NPC. Candidate miRNAs were screened by microarray analysis and validated by real-time quantitative polymerase chain reaction (RT-qPCR). In the follow-up cohort including 102 patients, blood samples (plasma) were collected before the treatment initiation, 3 months, 6 months, and 12 months after treatments, and at the time of any recurrence or metastasis. Among these plasma miRNAs, miR-9-3p, miR-124-3p, miR-892b, and miR-3676-3p were significantly upregulated (P = 0.018, P = 0.039, P = 0.001, and P = 0.01, resp.) after treatment compared with pretreatment, and the four plasma miRNAs were downregulated again at recurrence or metastasis (P < 0.001, P = 0.015, P = 0.003, and P = 0.026, resp.). The dynamic changes in plasma miRNAs after treatment reflect the outcome of the disease and have the potential to monitor recurrence and metastasis in patients with NPC.

Immunological Stability ofClostridium difficileToxins in Clinical Specimens

2018 ◽  
Vol 39 (4) ◽  
pp. 434-438 ◽  
Author(s):  
Donna M. Schora ◽  
Lance R. Peterson ◽  
Elena A. Usacheva
Keyword(s):  
Clostridium Difficile ◽  
Detrimental Effect ◽  
Quantitative Polymerase Chain Reaction ◽  
Storage Conditions ◽  
Toxin Detection ◽  
Discernible Effect ◽  
Stool Specimens ◽  
Polymerase Chain ◽  
The Impact ◽  
Good Detection

OBJECTIVEThe impact of storage on stability and detection ofClostridium difficiletoxins in feces is poorly understood. The objective of this study was to investigate the immunological stability ofC. difficiletoxins in clinical stool specimens under different storage conditions by evaluating this stability using toxin detection by enzyme immunoassay (EIA).METHODSStool specimens positive forC. difficileinfection (CDI) by quantitative polymerase chain reaction (qPCR) were used for EIA testing with theC. difficileTox A/B II kit. The EIA-positive specimens were stored aerobically under refrigerated (4–10°C) and frozen (−30°C and −80°C) conditions. Measurement of toxin quantity was conducting using optical density (OD) on days 0, 14, 30, 60, 90, and 120 of storage.RESULTSClostridium difficiletoxins demonstrated good detection in undiluted stool specimens by EIA up to 120 days of storage. Good detection of the toxins was observed in diluted samples at refrigerated and −80°C temperatures. Dilution detrimentally affected toxin detection at −30°C.CONCLUSIONStorage of undiluted clinical stool specimens at refrigerated, −30°C, and −80°C temperatures for up to 120 days has no discernible effect on the immunological stability ofC. difficilecytotoxins. However, storage at −30°C has a detrimental effect onC. difficiletoxin stability in diluted specimens.Infect Control Hosp Epidemiol2018;39:434–438

Current status of mycotoxigenic fungi in cereal grains in the central region of Botswana: a mini survey

2020 ◽  
Author(s):  
Basadi Masitha ◽  
Bokani Sereme-Mothobole ◽  
Kago Kabelo
Keyword(s):  
Animal Feed ◽  
Storage Conditions ◽  
Cereal Grains ◽  
Current Status ◽  
Number Of Factors ◽  
Cereal Product ◽  
Mycotoxigenic Fungi ◽  
Alternaria Species ◽  
Polymerase Chain ◽  
And Storage

Mycotoxins are secondary metabolites produced by fungi that can contaminate food, both human and animal feed at all stages of the food chain. A number of factors play a role in the proliferation of mycotoxins such as climate, humidity, harvest and storage conditions. This study was looking at the occurrence and identification of the fungi obtained from the cereal grains in the central district of Botswana. Samples collected were yellow maize (18), white maize (4), millet (10), cowpeas (11), sorghum (11) and china peas (1) each weighing about 500 g. Upon the arrival of samples, water activities of the samples were obtained. Seeds were sterilized in sodium hypochlorite, to be plated onto PDA for fungal extraction. The polymerase Chain reaction was used for the identification of the fungi and samples were sent to Inqaba laboratories for sequencing. The results showed that yellow maize was contaminated by Fusarium, A. niger and A. flavus; white maize was contaminated by F. proliferatum, F. fujikuroi and Gibberella moniliformis; red sorghum was contaminated by A. flavus, A.oryzae, Penicillium, Alternaria and Chaetomium muelleri; millet was contaminated by Epicoccum sorghinum and curvularia branchyspora and cowpeas were contaminated by Aspergillus and Alternaria species. Overall the most contaminated cereal product was millet, yellow maize, white maize, cowpeas and red sorghum at 40%, 37%, 27%, 10% and 4% respectively.

scholarly journals Effects of Storage Conditions on Postharvest Qualities in Garlic Bulbs (Allium sativum L.)

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 805C-805
Author(s):  
Sun-Tay Choi ◽  
Ro-Na Bae* ◽  
Dae-Sung Chung ◽  
Seung-Koo Lee
Keyword(s):  
Weight Loss ◽  
Low Temperature ◽  
Pyruvic Acid ◽  
Room Temperature ◽  
Allium Sativum ◽  
Storage Period ◽  
Storage Conditions ◽  
Controlled Atmosphere ◽  
Atmosphere Storage ◽  
And Storage

To investigate quality changes of garlic associated with cultivars and storage conditions, northern type `Seosan' and sub-tropical type `Daeseo' garlics were stored at controlled atmosphere (O2 3%, CO2 5%, -1 ± 1°C) condition, low temperature (-1 ± 1°C), and room temperature (20 ± 5°C). The rate of sprouting, weight loss, enzymatic pyruvic acid content, and degree of greening in crushed garlic were determined during storage. The rate of sprouting was higher in `Daeseo' than in `Seosan' garlic in all storage conditions. Sprouting was effectively suppressed in low temperature and controlled atmosphere storage. Weight loss in `Daeseo' garlic was higher than in `Seosan' garlic. Enzymatic pyruvic acid (EP) contents increased for 3 months storage period, and then decreased gradually as the storage period was prolonged at room or low temperatures. However, EP content decreased dramatically during storage under CA condition in both cultivars. When garlic bulbs were crushed, greening appeared in the garlic stored at low temperature for more than one month. However, greening did not occur in the crushed garlic bulbs stored in CA condition.

Effects of Flavorings, Storage Conditions, and Storage Time on Survival of Staphylococcus aureus in Sürk Cheese

2005 ◽  
Vol 68 (7) ◽  
pp. 1487-1491 ◽  
Author(s):  
TUĞRUL M. MASATCIOĞLU ◽  
YAHYA K. AVŞAR
Keyword(s):  
Staphylococcus Aureus ◽  
Room Temperature ◽  
Storage Time ◽  
Storage Conditions ◽  
Black Pepper ◽  
Ash Content ◽  
Mold Growth ◽  
Storage Duration ◽  
And Storage ◽  
Over Time

The objectives of this study were to determine the cumulative effects of flavorings (chili pepper, thyme, mint, cumin, nutmeg, allspice, clove, cinnamon, black pepper, salt, and hot red pepper paste), storage conditions, and storage time on the survival of Staphylococcus aureus in Sürk cheese and to monitor the associated chemical changes. Sürk cheese, a traditional Turkish cheese, was produced by heating diluted nonfat yogurt and adding flavorings to the resultant acid-heat curd. The cheese was later inoculated with S. aureus, shaped conically, and stored aerobically for mold growth and anaerobically in olive oil for 30 days at room temperature. The moisture content of aerobically stored cheese decreased over time and led to increases in total solids, salt, salt-in-moisture, and ash content during ripening (P &lt; 0.05). The presence or absence of the flavorings had no significant effect, whereas storage conditions and storage duration decreased the survival of S. aureus (P &lt; 0.05).

scholarly journals Directions of Colour Changes of Nectar Honeys Depending on Honey Type and Storage Conditions

2015 ◽  
Vol 59 (2) ◽  
pp. 51-61 ◽  
Author(s):  
Allna Piotraszewska-Pająk ◽  
Anna Gliszczyńska-Świgło
Keyword(s):  
Room Temperature ◽  
Quality Criteria ◽  
Storage Conditions ◽  
Room Temperature Storage ◽  
Long Term Storage ◽  
Colour Parameters ◽  
Colour Changes ◽  
Important Quality ◽  
And Storage ◽  
Temperature Storage

AbstractThe colour of honey is one of the most important quality criteria for consumers. The colour depends mainly on the content of plant pigments but the honey consistency, shape, and size of the crystals may also influence the honey colour parameters. It is related to the crystallisation and decrystallisation processes of honey during storage. In the present study, directions of colour changes of honey during storage were evaluated using a tristimulus colorimeter and the CIE 1976 L*a*b* and CIE L*C*hosystems. The effect of time (3 and 9 months) and storage conditions (cold storage, room temperature storage with access to light, and room temperature storage without access to light) on the colour of nectar honeys was investigated. The results obtained showed that both the type of honey and the storage conditions influenced the honey colour parameters. Significant differences in direction and intensity of the colour changes of honey during storage were observed. These differences make it difficult to indicate which storage conditions are optimal to preserve the colour of the honey. It was found that acacia and heather honeys were the most susceptible to colour changes during long-term storage in all of the study’s applied conditions, whereas rape and buckwheat honeys were the most stable in colour parameters.

scholarly journals Effect of Storage Conditions on Quality and Shelf Life of Pumpkin Cookies

2020 ◽  
pp. 70-79
Keyword(s):  
Chemical Analysis ◽  
Environmental Conditions ◽  
Room Temperature ◽  
Storage Stability ◽  
Storage Conditions ◽  
Packaging Material ◽  
Spread Ratio ◽  
Physico Chemical ◽  
And Storage ◽  
Quality And Shelf Life

The chemical, physical evaluation and storage stability of cookies was carried out. studies on quality was based on physico-chemical analysis that is weight, diameter, thickness ,spread ratio, moisture, fat, protein, ash, crude fiber, carbohydrate content as well as sensory characteristics which was determined for fresh and stored sample. The characteristics of cookies were influenced by packaging material, environmental conditions and constituents present in flour. Cookies was packed in LDPE bags and stored at room temperature. This study was conducted at the interval of 15 days up to 45days.

scholarly journals Effects of Various Handling and Storage Conditions on Stability of Treponema pallidum DNA in Cerebrospinal Fluid

1998 ◽  
Vol 36 (7) ◽  
pp. 2117-2119 ◽  
Author(s):  
A. V. Villanueva ◽  
R. P. Podzorski ◽  
M. P. Reyes
Keyword(s):  
Central Nervous System ◽  
Cerebrospinal Fluid ◽  
Nervous System ◽  
Room Temperature ◽  
Treponema Pallidum ◽  
Storage Conditions ◽  
Freeze Thaw ◽  
Target Dna ◽  
The Central Nervous System ◽  
And Storage

Treponema pallidum DNA from even small numbers of organisms was detectable in cerebrospinal fluid (CSF) stored at room temperature or at 4°C for several hours and in CSF subjected to three freeze-thaw cycles. These results suggest that negative PCR results forT. pallidum from patients diagnosed with T. pallidum invasion of the central nervous system are probably not due to the loss of target DNA prior to testing.

Testing achene germinationof Potamogeton praelongus Wulfen

2013 ◽  
Vol 8 (1) ◽  
pp. 78-86 ◽  
Author(s):  
Romana Prausová ◽  
Jana Janová ◽  
Lenka Šafářová
Keyword(s):  
Room Temperature ◽  
Germination Rate ◽  
Storage Conditions ◽  
Natural Process ◽  
Anaerobic Conditions ◽  
Mechanical Disruption ◽  
Breaking Dormancy ◽  
Petri Dishes ◽  
And Storage ◽  
Uva Radiation

AbstractThe goal of this work was to determine the best method of breaking the achene dormancy in Potamogeton praelongus Wulfen. The ways of breaking achene dormancy studied in this experiment included methods of achene storage, stratification, UVA radiation, anaerobic conditions, mechanical disruption of achenes?outer layers and their chemical disruption by NaClO. Nine different treatments of achenes were combined with two methods of achene storage. Particular achene treatments and storage conditions were proven to have a significant impact on breaking dormancy. Although the highest germination rate (83.3%) was achieved when the dormancy was broken chemically by long effect of 100% concentrations of Savo detergent (containing 5% NaClO), the growth of the sprouts was subsequently inhibited due to toxic effects of Savo. Thus the most successful treatment was based on changing temperature, e.g. 2.5 months of cold storage followed by 14 days at room temperature (germination rate 32.7%). This treatment was also most similar to the natural process. Germinated achenes were also found in Petri dishes exposed to UVA radiation, anaerobic conditions and chemical disruption of the outer layers. Results of these treatments were influenced by the storage method.

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