Adenine

Manufactured by Merck Group

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Adenine is a nucleic acid base that is a fundamental component of DNA and RNA molecules. It serves as a building block for the genetic material in living organisms. Adenine plays a crucial role in various biological processes, including energy transfer and signal transduction.

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Lab products found in correlation

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328 protocols using adenine

Yeast media composition for respiratory and non-respiratory growth

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Rich medium (Yeast peptone Dextrose, YPD) consisted of 1% yeast extract (Thermo Fisher Scientific, product number 212750), 2% peptone (Thermo Fisher Scientific, product number 211677) and 50 mg/l adenine (Merck, product number A9126-100G), and was supplemented with appropriate carbon source [2% glucose (non-respiratory medium) or 3% glycerol (respiratory medium)], as indicated. Standard minimal medium (SMM) consisted of 0.67% yeast nitrogen base without amino acids (Merck, product number Y0626), supplemented with all standard amino acids [bought from Sigma (now Merck)] at 76 mg/l except leucine, which was added to 380 mg/l; adenine to 19 mg/l, and PABA to 7.6 mg/l, and 2% glucose was added as a carbon source⁴⁷.
For induction of PDT degradation, the composition of SMM was the same as described above, with the exception of adenine and glucose, which varied depending on the condition –“un-inducing” medium contained 50 mg/l adenine and 2% glucose, while “inducing” medium contained 2 mg/l adenine and 1% glucose.
For experiments when mitochondrial DNA (mtDNA) was visualized by Sybr Green I staining, SMM was supplemented with 340 mg/l isoleucine, 550 mg/l of leucine, and 430 mg/l of valine, to prevent parsing of nucleoids^{48 (link)}.

Sanyal S., Kouznetsova A., Ström L, & Björkegren C. (2024). A system for inducible mitochondria-specific protein degradation in vivo. Nature Communications, 15, 1454.

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Adenine-induced Chronic Renal Failure in Rats

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Animal experiments were performed as previously described (23 ). A total of 40 male Wistar rats (age, 6-7 weeks; weight, 160-180 g) were obtained from Nanjing Jiancheng Bioengineering Institute, and maintained in a 12 h light/dark cycle, with 50-60% humidity at 22-26˚C. All rats were provided ad libitum access to a standard diet and water. Rats were randomly divided into two groups; a control (n=20) and CRF (n=20) groups after 1 week of adaptive feeding. Adenine (2.5 g; Sigma-Aldrich; Merck KGaA) was added to 100 ml normal saline to prepare a 2.5% Adenine suspension. Rats in the CRF group received 250 mg/kg Adenine once a day via oral gavage for a total of 14 days, and Adenine was administrated every other day for the next 14 days. Rats in the control group received the same amount of normal saline. All rats were fasted for 12 h prior to the last administration. Rats were anesthetized with 2% sodium pentobarbital (50 mg/kg) 1 h after the final administration, and cervical dislocation of the spine was immediately performed following collection of 4-6 ml blood from the abdominal aorta. All animal experiments were approved by the Experimental Animal Center of Lianyungang Hospital of Traditional Chinese Medicine (Lianyungang, China; approval. no. IACUC-20200312-07).

Huo C., Wang L., Wang Q., Yang Y, & Chen B. (2021). Hydroxysafflor Yellow A inhibits the viability and migration of vascular smooth muscle cells induced by serum from rats with chronic renal failure via inactivation of the PI3K/Akt signaling pathway. Experimental and Therapeutic Medicine, 22(2), 850.

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Adenine-Induced CKD Rat Model

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The rats were kept under observation for ten days prior to the start of the experiment. They were then randomly divided into two groups, that is, normal controls (Nor group, n = 20) and CKD rats with vascular calcification (CKD group, n = 35). Adenine (Sigma Chemical Co., St. Louis, MO, USA) was dissolved in distilled water to prepare the Adenine suspension. The rats in the CKD group were fed with a 1.8% high-phosphorus diet and given 2.5% Adenine suspension (220 mg/kg/d) by gavage daily for 4 weeks and then every other day for the next 4 weeks. The rats in the Nor group were fed with standard chow and intragastrically given the same volume of saline at the same frequency. During the experiment, food and water intake by the rats, body weight, and general conditions of the rats were monitored.

Wei X., Wu W., Li L., Lin J., Liu Q., Gan L, & Ou S. (2018). Bone Morphogenetic Proteins 2/4 Are Upregulated during the Early Development of Vascular Calcification in Chronic Kidney Disease. BioMed Research International, 2018, 8371604.

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Dietary Phosphorus Modulation in CKD Mice

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Female mice were used in the calcification studies, because they have shown higher susceptibility to calcification than male mice.25 Fourteen‐week‐old control mice and SM‐IκBΔN mice were fed a normal phosphorus diet (NPD: 1.0% calcium and 1.0% phosphorus; CLEA Japan, Tokyo, Japan) or a high phosphorus diet (HPD: 1.0% calcium and 2.0% phosphorus, CLEA Japan) ad libitum for 10 weeks. Some of these mice were also fed a diet containing 0.25% adenine (Sigma, St. Louis, MO) for 5 weeks (from 14 to 19 weeks of age) to induce CKD. A subset of the HPD‐fed/adenine‐fed CKD mice were treated for 10 weeks with one of the following procedures: 3% CaCO₃ in dietary chow, 3 mmol/L tempol (4‐hydroxy‐TEMPO, Sigma) in drinking water, or intraperitoneal injections of triptolide (70 μg/kg body weight, Sigma) every other day. The tempol‐containing water was changed every other day. At the end of the experiment, tissues including the aorta and the heart, as well as blood samples, were collected. Tissues from some mice were directly fixed in 4% paraformaldehyde to obtain whole cardiovascular images using Alizarin red staining, while other tissues were cut into multiple pieces for histology, immunostaining, RNA analysis, and measurement of the calcium content.

Yoshida T., Yamashita M., Horimai C, & Hayashi M. (2017). Smooth Muscle–Selective Nuclear Factor‐κB Inhibition Reduces Phosphate‐Induced Arterial Medial Calcification in Mice With Chronic Kidney Disease. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 6(11), e007248.

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Adenine-Induced Ferroptosis in PTEC

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PTEC were seeded (100,000 cells/well in DM) in 24-well flat-bottom plates to allow overnight adherence and then further cultured for 48 h (unless otherwise specified) in fresh DM in the absence (0 mM) of adenine (0.002 M sodium hydroxide (NaOH) vehicle control) or presence of adenine (2 mM or 8 mM in 0.002 M NaOH solution) (Sigma-Aldrich). In selected experiments, PTEC were stimulated with 5 µM erastin (MedChemExpress, Monmouth Junction, NJ, USA) for 48 h as a positive control for ferroptosis induction.
For inhibitor studies, 1 µM baicalein (Sigma-Aldrich), 10 µM ferrostatin-1 (Sigma-Aldrich) or 0.033% dimethyl sulfoxide (DMSO) vehicle control were added for the final 24 h of the treatment period. PTEC were harvested by trypsin treatment and analysed for protein expression by Western blotting. PTEC mitochondrial superoxide levels, mitochondrial function and necrosis were assessed by flow cytometry, with cell acquisition performed on an LSR Fortessa (BD Biosciences, San Jose, CA, USA) and data analyzed with FlowJo software (TreeStar, Ashland, OR, USA).

Khan M.A., Nag P., Grivei A., Giuliani K.T., Wang X., Diwan V., Hoy W., Healy H., Gobe G, & Kassianos A.J. (2022). Adenine overload induces ferroptosis in human primary proximal tubular epithelial cells. Cell Death & Disease, 13(2), 104.

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Adenine-Induced CKD Model in C57BL/6J Mice

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Adult (10–20 weeks old) male C57BL/6J (National Laboratory Animal Center, NLAC, Taipei, Taiwan) and PER2::LUC [6 (link)] under C57BL/6J background (Jackson Laboratory, Bar Harbor, ME) were maintained in a temperature and humidity controlled animal room (breeding: 21.8 ± 0.4 °C, 59 ± 3%; experiment: 20.5 ± 0.4 °C, 57 ± 5%; mean ± SD) under a 12:12 light:dark (L:D) cycle (light on at 7:00 h; off at 19:00 h) before being transferred to constant darkness (DD) for monitoring of circadian freerunning rhythm. All mice were habituated to a powdered normal diet (LabDiet, St. Louis, MO, USA) for one week prior to adenine diet feeding. A 0.2 % (w/w) adenine diet was prepared by adding 2 g of adenine (Sigma-Aldrich, St. Louis, MO, USA) to 1,000 g of the powdered diet. To create the adenine-induced CKD model, mice were given the adenine diet for 5–6 weeks [22 (link)]. Control mice were continuously fed with powdered normal diet. All animal protocols used in this study were approved by the Institutional Animal Care and Use Committee (IACUC) of Taipei Medical University Laboratory Animal Center. IACUC protocol numbers were: LAC-2017-0264 (approval date: November 14, 2017), LAC-2017-0469 (9 February 2018), and LAC-2018-0369 (3 January 2019).

Myung J., Wu M.Y., Lee C.Y., Rahim A.R., Truong V.H., Wu D., Piggins H.D, & Wu M.S. (2019). The Kidney Clock Contributes to Timekeeping by the Master Circadian Clock. International Journal of Molecular Sciences, 20(11), 2765.

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Adenine-Induced CKD-MBD Model in Mice

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Adenine model of CKD-MBD was established with C57BL/6 male mice of 8-weeks of age according to a previously-established protocol33 (link). Mice were randomly divided into Control, TSA (SelleckChem, USA), Adenine (Sigma-Aldrich, USA) and TSA-treated Adenine groups (n = 6) and the experiments went for 6 weeks. Control mice were fed a standard powder diet containing 1.16% calcium, 0.73% phosphate, 18.2% protein, and 7.56 IU/g vitamin D3, (Collaboration BioMedical Inc., Nanjing, China); TSA mice received intraperitoneal injection of TSA (0.5 mg/kg body weight in 100 μl of PBS) daily; Adenine mice were fed the regular diet containing 0.2% Adenine. At sacrifice, kidney, blood and femur were collected and stored at –80 °C until analysis. Use of animal and the experimental protocol were in accordance with the University Guidelines and approved by the Institutional Animal Care Committee (IACUC) of Nanjing University Medical School.

Lin W., Li Y., chen F., Yin S., Liu Z, & Cao W. (2017). Klotho preservation via histone deacetylase inhibition attenuates chronic kidney disease-associated bone injury in mice. Scientific Reports, 7, 46195.

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Adenine-Induced Chronic Renal Failure in Rats

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A total of 30 eight-week-old male Wistar rats (200-220 g) were purchased from Shanghai SLAC Laboratory Animal Company Limited (Shanghai, China). Rats in the CRF group were given 250 mg/kg adenine (Sigma-Aldrich, St. Louis, MO) by oral gavage once daily for 14 days, and these rats were administrated with adenine every other day for the next 14 days. The sham group was given the same amount of normal saline. For the Hiru-treated group, rats were intraperitoneally injected with different doses of Hiru (10, 20, and 40 IU/kg), respectively [18 ]. Subsequently, rats were euthanized at Day 0 (D0), D7, D14, and D28, respectively. The blood samples and arterial tissues were collected for subsequent experiments. The animal study was approved by the Animal Research Ethics Committee of Lianyungang Hospital of Traditional Chinese Medicine.

Chen B., Ding X, & Yang Y. (2022). Hirudin Regulates Vascular Function in Chronic Renal Failure through Modulating Macrophage Polarization. BioMed Research International, 2022, 6043698.

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Culturing Human Skin Epithelial Cells

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Human skin epithelial cells (HSEC) (CRL-4048) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA). Cells were cultured in epithelial cell culture medium consisting of a mixture of HAM-F12 (150 mL), DMEM (300 mL) and FBS (50 mL) supplemented with penicillin/streptomycin (50 IU/mL), adenine (24 µg/mL), insulin (5 µg/mL), triiodothyronine (1.3 ng/mL), hydrocortisone (0.4 µg/mL) and epidermal growth factor (EGF) (10 mg/mL) (all from Sigma-Aldrich/Merck). The HSEC were grown at 37 °C in a humidified incubator with 5% CO₂, and the culture medium was replaced every 2–3 days.

Cases-Perera O., Blanco-Elices C., Chato-Astrain J., Miranda-Fernández C., Campos F., Crespo P.V., Sánchez-Montesinos I., Alaminos M., Martín-Piedra M.A, & Garzón I. (2022). Development of secretome-based strategies to improve cell culture protocols in tissue engineering. Scientific Reports, 12, 10003.

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Analytical Characterization of Meteorite Sample NWA 1465

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Formamide, D-ribose, 2-D-deoxyribose and adenine were purchased from Aldrich. NWA 1465 was obtained from Sahara-nayzak, Asnieres sur Seine, France. Gas-chromatography mass-spectrometry (GC-MS) analyses were performed with a LC/GG MS Combo Agilent and Shimadzu GC-MS QP5050A with a Variant CP8944 column (WCOT fused silica, film thickness 0.25 μm, stationary phase VF-5ms, Øί 0.25 mm, lenght 30 m). UHPLC were performed on Ultimate 3000 Rapid Resolution system (DIONEX, Sunnyvale, USA) using Reprosil C18 column (2,5 μm × 150 mm × 2.0 mm). MALDI were performed on Q-Exactive (Thermo). NMR spectra were acquired with a Bruker Avance III 500 spectrometer. Computations were carried out at B3LYP/6-31 + G* level within the COSMO continuum solvent approximation.

Saladino R., Bizzarri B.M., Botta L., Šponer J., Šponer J.E., Georgelin T., Jaber M., Rigaud B., Kapralov M., Timoshenko G.N., Rozanov A., Krasavin E., Timperio A.M, & Mauro E.D. (2017). Proton irradiation: a key to the challenge of N-glycosidic bond formation in a prebiotic context. Scientific Reports, 7, 14709.

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