scholarly journals Acute Effects of Whole Egg Consumption on Circulating MicroRNAs in Sprague Dawley Animals (P15-001-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Amanda Bries ◽  
Joe Webb ◽  
Paige Curry ◽  
Savina Fischer ◽  
Elizabeth McNeill ◽  
...  
Keyword(s):  
Weight Gain ◽  
Corn Oil ◽  
Rna Extraction ◽  
Regulation Of Gene Expression ◽  
Circulating Microrna ◽  
Sprague Dawley ◽  
Total Rna ◽  
Rna Integrity ◽  
Zdf Rats ◽  
Whole Egg

Abstract Objectives MicroRNAs, small ∼22 nt non-coding RNAs, have a functional role in post-transcriptional regulation of gene expression. We have shown that whole egg-based diets reduce weight gain despite increased food intake compared to casein-based diets in Zucker-Diabetic Fatty (ZDF) rats. We hypothesized that the microRNA content of whole egg may underlie the differential impact on weight gain in ZDF rats. Our first objective was to characterize the circulating microRNA content in response to feeding whole egg-based diets over an 8 hour period. Our second objective was to identify and characterize the microRNAs present in whole egg- and casein-based diets. Methods Male Sprague Dawley rats (n = 12; 6 weeks of age) were meal-trained to consume 4 grams of feed in one hour. After one week of meal-training, animals were fasted overnight for 10 hours (h) with water ad libitum followed by controlled feeding of either an AIN93G casein-based diet or isocaloric AIN93G diet containing whole egg (20% egg protein, w/w). At 8 weeks of age, serum was collected via the tail vein at 0, 2, 4, 6, and 8 h following refeeding. Serum microRNA was extracted using a RNA SPLIT kit and prepared for sequencing on an Illumina HiSeq 3000 using a smallRNA Library Prep. Five RNA extraction methods were tested on 6 samples of each diet: 1) Trizol (Invitrogen), 2) Trizol + Directzol (Zymogen), 3) Trizol followed by a second extraction using a fatty acid RNA extraction kit (Norgen), 4) mirPremier (Sigma-Aldrich), 5) ISOLATE II RNA (Bioline). RNA 260/280 ratios were assessed via Nanodrop and RNA integrity was assessed on an Agilent Bioanalyzer. Results The greatest total RNA recovery (ng/µL) and most accurate 260/280 absorbance ratios were achieved using Trizol. The following concentrations of extracted RNA and their respective ratios (means ± SD) were achieved for each dietary component: casein diet (460.9 ± 528.1; 1.56 ± 0.12) and whole egg diet (394.3 ± 292.8; 1.55 ± 0.16), whole egg (73.4 ± 20.3; 1.15 ± 0.23), corn oil (67.5 ± 20.0; 1.41 ± 0.04), and cornstarch (247.12 ± 110.34; 1.64 ± 0.13). Total RNA recovered from the diets had an RNA integrity number (RIN) of 0 and microRNA concentrations were undetected. Conclusions Despite successful total RNA recovery from the diets, all samples had a RIN value of 0, which is not optimal for downstream sequencing. Currently, microRNAs from plasma are being sequenced to determine if varying diets had differential effects on circulating microRNA expression. Funding Sources Presidential Interdisciplinary Research Seed Grant Program, Iowa State University.

scholarly journals Effect of RNA preservation methods on RNA quantity and quality of field collected avian whole blood

2021 ◽  
Author(s):  
Johanna A Harvey ◽  
Sarah A Knutie
Keyword(s):  
Active Material ◽  
Rna Extraction ◽  
Quality Measures ◽  
Rna Degradation ◽  
Blood Samples ◽  
Dry Ice ◽  
Total Rna ◽  
Rna Integrity ◽  
Avian Blood ◽  
Rna Preservation

A limitation of comparative transcriptomic studies of wild avian populations continues to be sample acquisition and preservation to achieve resulting high-quality RNA (i.e., ribonucleic acids that transfers, translates, and regulates the genetic code from DNA into proteins). Field sampling of wild bird samples provides challenges as RNA degradation progresses quickly and because cryopreservation is often not feasible at remote locations. We collected blood samples from songbirds, as avian blood is nucleated and provides sufficient transcriptionally active material in a small and non-lethal sample, to compare the efficacy of widely available RNA stabilizing buffers, RNAlater (Ambion) and DNA/RNA Shield (Zymo) at differing concentrations along with a dry ice-based flash freezing method (Isopropanol 99% and dry ice mixture, -109°C). Each blood sample was divided among five different preservation treatments (dry ice-based flash freezing, RNAlater with 1:5 or 1:10 dilution, or DNA/RNA Shield with 1:2 or 1:3 dilution). A new protocol was optimized for total RNA extraction from avian blood samples with small starting volumes enabling sampling of small passerines. We quantified quality measures, RNA integrity numbers (RINe), rRNA ratios, and total RNA concentrations. We found that RNA preservation buffers, RNAlater and DNA/RNA Shield at all concentrations, provide sample protection from RNA degradation. We suggest caution against using dry ice-based flash-freezing alone for samples preservation as these samples resulted in lower quality measures then samples in preservation buffer. Total RNA concentration was generally not affected by preservation treatment and may vary due to differences in initial samples volumes and carryover across processing steps.

scholarly journals Dietary Whole Egg Reduces Body Weight Gain in a Dose-Dependent Manner in Zucker Diabetic Fatty Rats

2019 ◽  
Vol 149 (10) ◽  
pp. 1766-1775 ◽  
Author(s):  
Cassondra J Saande ◽  
Joseph L Webb ◽  
Paige E Curry ◽  
Matthew J Rowling ◽  
Kevin L Schalinske
Keyword(s):  
Vitamin D ◽  
Body Weight ◽  
Weight Gain ◽  
Body Weight Gain ◽  
Dependent Manner ◽  
Dietary Recommendations ◽  
Zucker Diabetic Fatty Rats ◽  
Zdf Rats ◽  
Dose Dependent ◽  
Whole Egg

ABSTRACT Background We previously reported that a whole-egg–based diet attenuated weight gain in rats with type 2 diabetes (T2D) and more effectively maintained vitamin D status than an equivalent amount of supplemental cholecalciferol. Objectives The objective of this study was to determine the lowest dose of whole egg effective at maintaining vitamin D homeostasis and attenuating the obese phenotype in T2D rats. Methods Zucker diabetic fatty (ZDF) rats (n = 40; age 6 wk; prediabetic) and their lean controls (n = 40; age 6 wk) were randomly assigned to a diet containing 20% casein (CAS) or 20%, 10%, 5%, or 2.5% protein from whole egg (20% EGG, 10% EGG, 5% EGG, and 2.5% EGG, respectively). All diets contained 20% total protein (wt:wt). All rats received their respective diets for 8 wk, at a stage of growth and development that translates to adolescence in humans, until 14 wk of age, a point at which ZDF rats exhibit overt T2D. Weight gain was measured 5 d/wk, and circulating 25-hydroxyvitamin D [25(OH)D] was measured by ELISA. Mean values were compared by 2-factor ANOVA. Results The 20% EGG diet maintained serum 25(OH)D at 30 nmol/L in ZDF rats, whereas the 10%, 5%, and 2.5% EGG diets did not prevent insufficiency, resulting in mean serum 25(OH)D concentrations of 24 nmol/L in ZDF rats. Body weight gain was reduced by 29% (P < 0.001) and 31% (P < 0.001) in ZDF rats consuming 20% and 10% EGG diets, respectively, and by 16% (P = 0.004) and 12% (P = 0.030) in ZDF rats consuming 5% and 2.5% EGG diets, respectively, compared with CAS. Conclusions Whole-egg–based diets exerted a dose-dependent response with respect to attenuating weight gain. These data could support dietary recommendations aimed at body weight management in individuals predisposed to obesity and T2D.

scholarly journals Whole Egg Consumption in Zucker Diabetic Fatty Rats Display a Dose-dependent Reduction in Weight Gain and Total Body Fat, Accompanied by an Increase in Lean Body Mass (P21-054-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Joe Webb ◽  
Cassondra Saande ◽  
Kevin Schalinske ◽  
Matthew Rowling
Keyword(s):  
Weight Gain ◽  
Lean Body Mass ◽  
Body Mass ◽  
Body Fat ◽  
Egg Protein ◽  
Total Body Fat ◽  
Zdf Rats ◽  
Total Body ◽  
Dose Dependent ◽  
Whole Egg

Abstract Objectives The objective of this study was to determine the lowest dose of whole egg-based diets to effectively attenuate the obese phenotype in type 2 diabetic (T2D) rats using a dose-response experimental design. Methods Male Zucker diabetic fatty (ZDF) rats (n = 8) and their lean controls (n = 8) were obtained at 6 weeks of age. Following one week of acclimation, animals were randomly assigned to one of 5 treatment groups: a casein-based diet (20% protein, w/w) or a whole-egg based diet provided at either 20, 10, 5, or 2.5% egg protein (w/w). Animals were fed their respective diets for 8 weeks with weight gain and food intake measured daily. At 14 weeks of age, body composition was analyzed by dual X-ray absorptiometry and statistical differences were measured between groups using a 2-way ANOVA at P < 0.05. Results Whole egg-based diets exerted a dose-dependent decrease in cumulative body weight gain and final body weight; increased in food intake; decreased total body fat; and increased lean body mass. Interestingly, the 20% whole egg protein diet decreased body fat and increased lean body mass in the ZDF rats and their lean controls. Conclusions Together, these data support the hypothesis that dietary consumption of whole eggs may decrease weight gain, reduce body fat, and increase lean body mass in a dose-dependent manner in ZDF rats. These results suggest the need to modify dietary recommendations during T2D and obesity to potentially consume more whole egg. Funding Sources This work was supported by the Egg Nutrition Center and in part by a National Science Foundation Graduate Research Fellowship. Supporting Tables, Images and/or Graphs

scholarly journals PERBANDINGAN TIGA KIT EKSTRAKSI RNA UNTUK ANALISIS TRANSKRIPTOMIKA PADA KELAPA SAWIT (Elaeis guineensis Jacq.)

2019 ◽  
Vol 6 (1) ◽  
pp. 118
Author(s):  
Siti Zulaeha ◽  
Devit Purwoko ◽  
Imam Cartealy ◽  
Teuku Tajuddin ◽  
. Karyanti ◽  
...  
Keyword(s):  
Somatic Embryo ◽  
Oil Palm ◽  
Elaeis Guineensis ◽  
Early Stage ◽  
Rna Extraction ◽  
Transcriptomic Analysis ◽  
Total Rna ◽  
Rna Integrity ◽  
Rna Quality ◽  
Rna Integrity Number

Comparison of Three RNA Extraction Kits for Transcriptome Analysis of Oil Palm (Elaeis guineensis Jacq.) ABSTRACTObtaining high-quality RNA is very important at an early stage of molecular biology research. To isolate RNA, high skill and caution are required in following laboratory procedures because RNA is easily degraded, especially samples from plant tissue culture. One of the parameters used to check the total RNA quality is RIN (RNA Integrity Number). The aim of this study was to obtain RNA extraction methods on oil palm leaves, callus and somatic embryos that were of good quality and high concentrations for transcriptomic analysis. RNA extraction was carried out using Plant RNA PureLink (Ambion), Genezol RNA Extraction (Geneaid) and RibospinTM Plant (Geneall) kit methods. The results showed that oil palm leaf, callus and somatic embryo RNA were successfully extracted using the RibospinTM (Geneall) kit. Based on the total RNA number of more than 4 μg and the RIN value of more than 7, the extracted RNA could be used in RNA sequencing for transcriptomic analysis. Keywords: callus, oil palm, RNA analysis, RNA quality, somatic embryo ABSTRAKMenghasilkan RNA berkualitas tinggi sangatlah penting pada tahap awal penelitian biologi molekuler. Untuk mengisolasi RNA diperlukan keterampilan dan kehati-hatian tinggi dalam mengikuti prosedur di laboratorium karena RNA lebih mudah terdegradasi, khususnya sampel hasil kultur jaringan tanaman. Salah satu parameter yang digunakan pada pengecekan kualitas RNA total adalah RIN (RNA Integrity Number). Penelitian bertujuan mendapatkan metode ekstraksi RNA pada daun, kalus dan embrio somatik kelapa sawit yang berkualitas baik dan memiliki konsentrasi tinggi untuk analisa transkriptomika.  Ekstraksi RNA dilakukan menggunakan metode kit Plant RNA PureLink (Ambion), Genezol RNA Extraction (Geneaid) dan RibospinTM Plant (Geneall). Hasil menunjukkan bahwa RNA daun, kalus dan embrio somatik kelapa sawit telah berhasil diekstraksi dengan menggunakan kit RibospinTM (Geneall). RNA hasil ekstraksi tersebut dapat digunakan untuk sekuensing RNA dengan tujuan analisis transkriptomika, dilihat dari jumlah total RNA yang lebih dari 4 μg dan nilai RIN lebih dari 7. Kata Kunci: analisis RNA, embrio somatic, kalus, kelapa sawit, kualitas RNA 

Ultrastructural Effects of Acute Kepone Treatment in Rats

1976 ◽  
Vol 34 ◽  
pp. 286-287
Author(s):  
Richard L. Klein ◽  
Åsa K. Thureson-Klein ◽  
Harihara M. Mehendale
Keyword(s):  
Neurological Disorders ◽  
Corn Oil ◽  
Preliminary Investigation ◽  
Reproductive Capacity ◽  
Ultrastructural Changes ◽  
Lipid Deposition ◽  
Severe Toxicity ◽  
Sprague Dawley ◽  
Ether Anesthesia ◽  
Sprague Dawley Rats

KeponeR (decachlorooctahydro-1,3,4-metheno-2H-cyclobuta[cd]pentalen-2-one) is an insecticide effective against ants and roaches. It can cause severe toxicity in fishes, birds, rodents and man. Prominent effects include hepatic lipid deposition and hypertrophy, impairment of reproductive capacity and neurological disorders. Mitochondrial oligomycin-sensitive Mg2+-ATPase is also inhibited. The present study is a preliminary investigation of tissue ultrastructural changes accompanying physiological signs of acute toxicity, which after two days treatment include: pronounced hypersensitivity and tremor, various degrees of anorexia and adipsia, and decreased weight gain.Three different series of adult male Sprague-Dawley rats (Charles River or CD-I) were treated by intubation with Kepone in corn oil at a dose of 50 mg per kg for 3 successive days or at 200 ppm in food for 8 days. After ether anesthesia, rats were immediately perfused via a cannula in the left ventricle with 4% p-formaldehyde and 0.5% glutaraldehyde in Millonig's phosphate buffer at pH 7.2 for 20-30 min at 22°C.

Liver Toxicity in Rats Fed A Mixture of Pcb Congeners and 2,3,7,8-Tcdd

1997 ◽  
Vol 3 (S2) ◽  
pp. 51-52
Author(s):  
B.J. Cornell ◽  
A. Singh ◽  
I. Chu
Keyword(s):  
Aromatic Compounds ◽  
Liver Toxicity ◽  
Corn Oil ◽  
Morphological Changes ◽  
Test Substance ◽  
Sprague Dawley ◽  
Morphological Alterations ◽  
Pcb Congeners ◽  
Transmission Electron ◽  
Sprague Dawley Rats

Polyhalogenated aromatic compounds such as polychlorinated biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins (PCDDs) continue to be environmental contaminants because of their bioaccumulation in the food chain and high resistance to biodegradation. The current study was undertaken to determine if a mixture of PCB congeners (WHO-IPCS) were interactive with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in producing morphological changes in the rat liver. Both compounds are known to produce a broad range of biochemical and morphological alterations including enzyme induction.Groups (N=5) of female Sprague-Dawley rats were administered TCDD (0, 2.5, 25, 250, 1000 ng/kg bw/day) or PCB (0, 2, 20 μg/kg bw/day) alone, or in combination with each concentration of both compounds. Incorrect concentrations were published in a previous abstract. The test substance was mixed with corn oil and given by gavage at 2 ml/kg daily for 28 days. At the end of the experiment, the rats were killed and liver samples were prepared for transmission electron microscopy.

scholarly journals THE MODULATION OF DRUG EFFLUX TRANSPORTER BY CURCUMIN IN MCF7 BREAST CANCER CELLS AFTER REPEATED EXPOSURE OF ENDOXIFEN AND ESTRADIOL

2018 ◽  
Vol 10 (1) ◽  
pp. 102
Author(s):  
Robby Hertanto ◽  
Wilson Bastian ◽  
Paramita . ◽  
Melva Louisa
Keyword(s):  
Breast Cancer ◽  
Cell Viability ◽  
Mrna Expression ◽  
Rna Extraction ◽  
Efflux Transporters ◽  
Drug Efflux ◽  
Mcf7 Cells ◽  
Total Rna ◽  
P Glycoprotein ◽  
Drug Efflux Transporters

Objective: The aim of the present study was to determine whether curcumin (CM) can prevent drug sensitivity of breast cancer (BC) cells when E andβ-E2 are administered together and whether the underlying mechanism involves modulation of drug efflux transporters.Methods: MCF7 BC cells were treated with the vehicle only, E+β-E2, or E+β-E2+CM repeatedly for 8 weeks. Afterward, the cells were harvested,counted, and isolated for total RNA extraction. Total RNA was then processed into cDNA and further processed for the determination of mRNAexpression patterns of drug efflux transporters (P-glycoprotein, BCRP, and MRP1).Results: Decreased sensitivity of BC cells was shown by the increased cell viability of MCF7 cells after 8 weeks. This condition was accompanied withincreased mRNA expression of P-glycoprotein, BCRP, and MRP1 in cells treated with E+β-E2, as compared with the vehicle only. CM, administered incombination with E+β-E2, resulted in decreased cell viability versus E and β-E2 and also decreased in mRNA expression of P-glycoprotein, BCRP, andMRP1.Conclusion: CM partially reversed the sensitivity loss of BC cells to E in the presence of β-E2 by modulating drug efflux transporters.

The prophylactic potential of Zingiber officinale flowers and leaves extract to mitigate hyperglycemia in Sprague Dawley rats

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Saira Tanweer ◽  
Tariq Mehmood ◽  
Saadia Zainab ◽  
Zulfiqar Ahmad ◽  
Muhammad Ammar Khan ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Body Weight ◽  
Weight Gain ◽  
Body Weight Gain ◽  
Zingiber Officinale ◽  
Sprague Dawley ◽  
Content Type ◽  
Sprague Dawley Rats ◽  
Hematological Analysis ◽  
Therapeutic Tools

Purpose Innovative health-promoting approaches of the era have verified phytoceutics as one of the prime therapeutic tools to alleviate numerous health-related ailments. The purpose of this paper is to probe the nutraceutic potential of ginger flowers and leaves against hyperglycemia. Design/methodology/approach The aqueous extracts of ginger flowers and leaves were observed on Sprague Dawley rats for 8 weeks. Two parallel studies were carried out based on dietary regimes: control and hyperglycemic diets. At the end of the experimental modus, the overnight fed rats were killed to determine the concentration of glucose and insulin in serum. The insulin resistance and insulin secretions were also calculated by formulae by considering fasting glucose and fasting insulin concentrations. Furthermore, the feed and drink intakes, body weight gain and hematological analysis were also carried out. Findings In streptozotocin-induced hyperglycemic rats, the ginger flowers extract depicted 5.62% reduction; however, ginger leaves extract reduced the glucose concentration up to 7.11% (p = 0.001). Similarly, ginger flowers extract uplifted the insulin concentration up to 3.07%, while, by ginger leaves extract, the insulin value increased to 4.11% (p = 0.002). For the insulin resistance, the ginger flower showed 5.32% decrease; however, the insulin resistance was reduced to 6.48% by ginger leaves (p = 0.014). Moreover, the insulin secretion increased to 18.9% by flower extract and 21.8% by ginger leave extract (p = 0.001). The feed intake and body weight gain increased momentously by the addition of ginger flowers and leaves; however, the drink intake and hematological analysis remained non-significant by the addition of ginger parts. Originality/value Conclusively, it was revealed that leaves have more hypoglycemic potential as compared to flowers.

Ultrastructural changes of Basolateral Amygdaloid nuclei in the Sprague Dawley rats brain following exposure to naphthalene balls

2021 ◽  
Vol 12 (2) ◽  
pp. 1272-1275
Author(s):  
Angu Bala Ganesh K S V ◽  
Sujeet Shekhar Sinha ◽  
Kesavi Durairaj ◽  
Abdul Sahabudeen K
Keyword(s):  
Structural Changes ◽  
Corn Oil ◽  
Chromatin Condensation ◽  
Ultrastructural Changes ◽  
High Dose ◽  
Model Group ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
The Brain ◽  
Amygdaloid Nuclei

Naphthalene is a bicyclic aromatic constituent commonly used in different domestic and marketable applications comprising soil fumigants, lavatory scent disks and mothballs. Accidentally, workers, children and animals are exposed to naphthalene mothballs, so there is a need to study the pathology behind this chemical toxicity. The current study was carried out to assess the ultra structural changes of basolateral amygdaloid nuclei in the Sprague Dawley rats brain in association to naphthalene toxicity. The toxicity model group was administered with naphthalene (200 and 400mg) using corn oil as a vehicle for 28 days. The post delayed toxicity of naphthalene high dose ingestion was also assessed in rats. After the experimental period, the brain tissue was processed to observe the ultra structural changes using a transmission electron microscope. The alterations in cell organelles, nuclei damage, mitochondrial swelling, chromatin condensation suggested naphthalene induced damage in the neurons of the basolateral amygdala of the brain in the toxicity model group. These experimental trials provide information about the alert of mothball usage in the home and identify risks linked with accidental exposure and misuse.

Sign in / Sign up

Export Citation Format

Share Document