A total of 15 virgin female mice were OVXed. Letrozole was prepared by dissolving in ethanol (8 mg/mL) followed by addition of 0.9% NaCl to a final concentration of 0.8 mg/mL. Ten days later, the mice were intraperitoneally injected with Letrozole (Novartis, Basel, Switzerland) at a dosage of 4 μg/g body weight, or vehicle as a control once per day for 4 days (Fig 1 , lower). Four hours after the final Letrozole treatment, behavior was analyzed as described in paradigm 1. Additionally, approach to pups was analyzed as in paradigm 2.
Letrozole
Manufactured by Novartis
Sourced in Switzerland, Germany, India
Letrozole is a selective aromatase inhibitor used in the treatment of breast cancer. It works by blocking the enzyme aromatase, which is responsible for the production of estrogen in the body. This can help slow the growth of certain types of breast cancer that are dependent on estrogen for their growth.
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Lab products found in correlation
Femara, by Novartis (2 mentions)
Distilled h2o, by Merck Group (1 mentions)
Raloxifene hydrochloride, by Merck Group (1 mentions)
Anastrozole, by AstraZeneca (1 mentions)
Lncap, by American Type Culture Collection (1 mentions)
Mda mb 453, by American Type Culture Collection (1 mentions)
Skbr3, by American Type Culture Collection (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
Aromasin, by Pfizer (1 mentions)
Arimidex, by AstraZeneca (1 mentions)
Exemestane, by Pfizer (1 mentions)
P70 s6 kinase, by Cell Signaling Technology (1 mentions)
Cobalt chloride cocl2, by Merck Group (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Actinomycin d, by Merck Group (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Trastuzumab, by Genentech (1 mentions)
Hif 1α, by BD (1 mentions)
4 androstenedione ad, by Merck Group (1 mentions)
Human insulin, by Merck Group (1 mentions)
17 protocols using letrozole
1
Letrozole-Induced Mouse Ovariectomy Model
2
Endocrine-Resistant Breast Cancer Cell Lines
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Endocrine-sensitive MCF-7 (American Type Culture Collection (ATCC) Virginia, USA ) and tamoxifen-resistant LY2 cells (kind gift from R. Clarke, Georgetown, DC, USA) were grown as previously described [21 (link)]. MCF7-derived AI-sensitive cells (Aro) were developed in house. MCF7 Aro-derived letrozole-resistant cells (LetR) and anastrozole-resistant cells (AnaR) were created by long-term treatment of Aro with letrozole (Novartis, Basel, Switzerland) or anastrozole (AstraZeneca, Macclesfield, UK) [21 (link)]. LY2, LetR and AnaR cells were utilised to model endocrine resistance developed through long-term adaptation to hormonal therapy. MDA-MB-453, SKBR3 and LNCaP cells were acquired from ATCC and maintained as recommended. Cells were maintained in steroid-depleted medium for 72 hours before treatment with hormones. All cells were incubated at 37 °C under 5 % CO2 in a humidified incubator. All in-house cells were authenticated and are routinely verified as endocrine resistant.
3
Letrozole-induced Obesity in Mice
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After one week of acclimatization, the mice were randomly divided into two groups (n = 10 each). The control animals were fed with a laboratory rodent diet (protein, 28.507%; carbohydrate, 57.996%; and fat, 13.496) for 20 weeks. The letrozole-treated mice were fed with 37.5 mg per kg letrozole (Femara, Novartis Pharma AG, Basel, Switzerland), dissolved in a laboratory rodent diet for an equivalent time. Body weights were measured weekly from 21 d to the end of the experiment. At the end of the experiment, the mice in both groups were sacrificed by decapitation. The whole-body fat distributions of both groups of mice were determined. The procedure of the experiments is Figure 11 .
4
Chondrocyte Response to Letrozole
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At the end of culture with serum, C-28/I2 and T/C-28a2 chondrocytes were changed to serum-free medium for 2 hours, cells were incubated alone or with a range of concentrations (10−11 M, 10−9 M and 10−7 M) of letrozole (Femara; Novartis Oncology, Nuremberg, Germany) during the two serum-free days (days 6 and 7 or days 8 and 9, respectively). For this purpose, stock solutions of 10−1 M letrozole (EtOH-soluble) were prepared and dissolved stepwise with distilled H2O (EMD Millipore, Billerica, MA, USA). All experiments were performed in triplicate, and controls were incubated with water instead of the aromatase inhibitor letrozole.
5
BIG 1-98 Randomized Breast Cancer Trial
6
Xenograft Mouse Model for Uveal Melanoma Treatment
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Forty-eight mice xenografted with mixed UM fragments from patient case 6 and 7 (Additional file 1 : Table S1) were equally grouped in to four groups according to the treatment protocols: control with 100 μl 0.9 % saline S.C. per week (n = 12) [35 (link)], AI (Letrozole; Femara® were prepared in 0.3 % hydroxypropylcellulose; mice were injected S.C. five times per week with Letrozole 10 μg per mouse per day; Novartis, Switzerland) (n = 12) [36 (link)], GnRHa (Leuprorelin acetate S.C. 10 mg/kg per week; LEUPLIN®DEPOT 3.75 mg S.C. Injection, Takeda, Japan) (n = 12) [37 (link)], and SERM (Raloxifene hydrochloride, 0.1 μg S.C. per day; Sigma-Aldrich, US) [32 (link)].
Laparotomic examination was performed and these mice were sacrificed 3 weeks after the xenograft procedures.
Laparotomic examination was performed and these mice were sacrificed 3 weeks after the xenograft procedures.
7
Letrozole Administration and Ovariectomy in Mice
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Letrozole was purchased from Novartis Oncology (Tokyo, Japan). Letrozole was suspended in distilled water (0.88 mmol/l). The experimental mice were divided into two groups containing ten mice each. The Letrozole group was given Letrozole 5 mg/kg/day by oral gavage every day until the end of the study, and the control group was given vehicle. Bilateral ovariectomy was performed under pentobarbital anesthesia in all experimental mice on the seventh day after commencement of Letrozole administration.
8
Letrozole Improves Survival in Sepsis
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To block aromatase activity, letrozole (provided by Novartis®, Basel, Switzerland) was administered daily subcutaneously at a dose of 10 µg [26 (link)] for 7 days prior to surgical procedures in groups of CLP and SS mice and continued afterward for the 5 days of observation. In control groups of CLP and SS mice, vehicle was administered subcutaneously daily as previously mentioned. Fifty mice underwent CLP with 30 receiving letrozole and 20 receiving vehicle. Thirty mice underwent SS with 20 receiving letrozole and 10 receiving vehicle. Survival or death and rating of illness severity were observed on a daily basis in all 80 mice over the 5 days of the protocol or until death (if death occurred prior to the fifth day of observation). However, in 17 mice that died before the end of the observation period (nine CLP with vehicle and eight CLP with letrozole), 29 observations of severity of illness were not recorded prior to death. Thus, severity of illness was not compared between groups unless reflected by death. In several of the animals that died, the hour of death was not clear. Thus, survival analysis was performed with mice censored if alive at day 5. The log-rank test was used to assess statistical significance of differences in survival curves. A P value of < 0.05 was considered significant.
9
Letrozole and Clomiphene Citrate Availability
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Letrozole (Novartis Pharmaceuticals, India) and clomiphene citrate (Aventis, India) tablets were purchased from a local pharmacy in Rajahmundry.
10
Adjuvant Endocrine Therapy and Diuretics
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Enrolled patients continued treated with the standard-dose of a third-generation AI; 1 mg Anastrozole (Arimidex; AstraZeneca Pharmaceuticals), 25 mg Exemestane (Aromasin; Pfizer), or 2.5 mg Letrozole (Femara; Novartis) orally daily. Patients who had been taking a stable dose of medication for treatment of pain (eg, nonsteroidal anti-inflammatory medications, cyclooxygenase-2 inhibitors and acetaminophen), and/or bisphosphonates every 6 month at the time of enrollment were permitted to continue the medication. Lasilactone® 50 mg; (produced by: Sanofi Aventis Egypt s.a.e. Under Licence of: Sanofi Aventis/ Germany) an oral combination of Frusemide 20 mg/Spironolactone 50 mg in a film-coated tablet, was coadministrated every other day with breakfast for 4 weeks. Patients were also instructed to avoid taking concomitant medications with the potential to interact with Lasilactone® as described in the package insert.
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