The chronic (CFA-induced inflammation) model was used for the assessment of the pharmacological effects of a CDPDP (20 mg/kg) and CDPDP (40 mg/kg) compared to ibuprofen. The Edema parameter was measured using a dial thickness gauge after inflammation was induced in the mouse paw (Mitutoyo Corporation, Japan).
Dial thickness gauge
Manufactured by Mitutoyo
Sourced in Japan
The Dial Thickness Gauge is a precision measuring instrument used to determine the thickness of objects. It features a dial that displays the measured thickness in a linear scale. The gauge is designed for accurate and reliable thickness measurements, making it a useful tool in various industrial and laboratory applications.
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19 protocols using dial thickness gauge
1
Anti-inflammatory Effects of CDPDP
2
Characterization of Pharmaceutical Tablets
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The prepared tablets were characterized for different post compression parameters such as weight variation, hardness, % friability, thickness, wetting time, drug content and disintegration time. The hardness of the tablets was assessed with the help of Monsanto hardness tester. It was done by taking the average value of any three tablets from every batch. Friability was tested by taking 20 tablets and placing the weighed tablets in the friability chamber (Roche friability tester) for 4 min at 25±1 rpm, and then percentage of the weight loss was determined. Disintegration test was performed in phosphate buffer of pH 6.8 at 37 ± 1 °C. 15 Thickness of tablets was determined using vernier caliper (Mitutoyo Dial Thickness Gauge, Mitutoyo, Japan). Wetting time was recorded by taking 10 ml of phosphate buffer pH 6.8 in petridish, over this fourfold tissue paper was placed and a tablet was carefully placed over this. Time required for a tablet to wet was recorded as wetting time. For determination of drug content ten tablets from each formulation batch was taken randomly and triturated to fine powder. Weight of powder equivalent to 150mg was taken in phosphate buffer pH 6.8 and assayed for drug content uniformity using double beam UV -visible spectrophotometer at λ max 325 nm.
3
Inducing Atopic Dermatitis in Mice
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The induction of AD using DFE was performed on the basis of our previous research.30 (link) The mice (n = 5) were divided into six groups, and the surfaces of both ear lobes were stripped very gently three times with surgical tape (Nichiban, Tokyo, Japan). Next, DFE (20 μL, 10 mg mL−1, dissolved in PBS plus 0.5% Tween 20, Greer laboratory Inc., Lenoir, NC, USA) was painted on each mouse ear twice a week for 3 weeks. One week after the initial treatment with DFE, which induced AD-like skin inflammation, the mouse ears were treated twice per week with CSN and MSN (100 or 500 μg per ear) by painting for the duration of the 3-week DFE treatment. Ear thickness was measured 24 h after DFE application using a dial thickness gauge (Mitutoyo, Co., Tokyo, Japan).
On day 21, the mice were euthanized with carbon dioxide. Whole blood samples were collected from the celiac artery. The serum was stored at −70℃ for further analysis. Subsequently, the ears were removed and histopathological analysis was performed. Total serum IgE and IgG2a levels were measured using an ELISA kit (BD Biosciences, Oxford, UK) according to the manufacturer’s instructions. For the detection of DFE-specific IgE, mite extract (10 mg) was coated in 96-well plates, and the DFE-specific IgE level was assessed by measuring the optical density of each well.
On day 21, the mice were euthanized with carbon dioxide. Whole blood samples were collected from the celiac artery. The serum was stored at −70℃ for further analysis. Subsequently, the ears were removed and histopathological analysis was performed. Total serum IgE and IgG2a levels were measured using an ELISA kit (BD Biosciences, Oxford, UK) according to the manufacturer’s instructions. For the detection of DFE-specific IgE, mite extract (10 mg) was coated in 96-well plates, and the DFE-specific IgE level was assessed by measuring the optical density of each well.
4
Contact Hypersensitivity Response Evaluation
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The mice were sensitised and challenged with PC according to a previously described method.19 Sensitisation was performed by application of 100 µL of 3% PC to their shaved abdomen. Negative control mice received acetone instead of PC. Six days later, the skin of the mice was challenged to elicit a contact hypersensitivity response by the application of 10 µL of 1% PC to the inner and outer surfaces of the right ear. Next, 20 µL of 1% LCZ, 1% LNF, 1% TBF, 1% AMO or vehicle (acetone) was applied to the same ear. The ear thickness was measured 24 hours after the PC application using a dial thickness gauge (Mitutoyo Corporation). Punch biopsies of 5‐mm diameter were then obtained from the skin of the right ear to determine the IFN‐γ content.
5
Hind Foot Measurements in Rat Models
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As previously described (18 (link), 19 (link)), dorsoplantar widths of the hind feet were measured using a Mitutoyo Corporation dial thickness gauge every 2-3 days from D11 until sacrifice. Right and left footpad widths from each rat were averaged, and then mean footpad widths from rats within each group were averaged and expressed as a mean ± standard error of the mean (SEM). The hind limbs were photographed under anesthesia.
6
Delayed Type Hypersensitivity Assay in Mice
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Delayed type hypersensitivity assay was performed as reported previously.15 (link) Briefly, to induce allosensitization, allogeneic BALB/c splenocytes (5 × 106 splenocytes) suspended in normal saline were subcutaneously injected into the nape of the neck. C57BL/6 mice were injected intravenously with Mock- or ALCAM-silenced MSCs (C57BL/6) on days 0 and 1 post-sensitization. To assess the generation of alloreactive Th1 generation, draining lymph nodes were harvested on day 7 for ELISPOT analysis. In another set of the experiment to measure ear-swelling, the mice were re-challenged, on day 14 post-sensitization, by epicutaneous injection of mitomycin-C treated BALB/c splenocytes (4 × 106 splenocytes) suspended in normal saline to the right ear. The left ear was injected with saline alone to serve as intra-mouse control. Ear thickness was measured pre-sensitization and at 24 and 48 h post-challenge using a dial thickness gauge (Mitutoyo, Kawasaki, Japan). The degree of swelling was calculated as the difference in thickness of the challenged ear (right ear) minus the baseline line thickness of the unchallenged ear (left ear). The mice were sacrificed 48 h post-challenge and the ear and draining lymph nodes were harvested for H&E and flow cytometry analysis.
7
Induction of Experimental Arthritis
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Mice were anaesthetised by isoflurane/O2/N2O and immunised intradermally (i.d.) with methylated bovine serum albumin (mBSA) (Sigma, St Louis, MO) emulsified in complete Freund's adjuvant (CFA) (Difco, Detroit, MI) at the tail base. Four days later mice were given 1000 µg (approx. 50 mg/kg body weight) anti-mouse type II collagen antibody (anti-CII) cocktail (Chondrex, Redmond, WA) containing the clones A2-10 (IgG2a), F10-21 (IgG2a), D8-6 (IgG2a), D1-2G (IgG2b) and D2-112 (IgG2b) intravenously in 200 µl phosphate-buffered saline (PBS). Seven days after immunisation the mice were challenged with 200 µg mBSA subcutaneously in 20 μl PBS in the right foot pad. The left foot pad was given 20 μl PBS only and served as control. Baseline paw and ankle measurements were made on the right paw on day 0 prior to mBSA challenge. Paw and ankle swelling was measured using a dial thickness gauge (Mitutoyo, Japan), and was calculated as right paw or ankle thickness minus baseline measurement.
8
Arachidonic Acid-Induced Ear Edema
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The ICR mice (n = 6) were orally pre-treated with SDE (40 mg/kg) or indomethacin (1 mg/kg) for 3 days. After the final treatment, arachidonic acid [2% (w/v)] was applied to the ear of the mouse (30 µl/ear). The ear thicknesses were measured using a dial thickness gauge (Mitutoyo, Japan), 1 h after the arachidonic acid treatment.
9
Thickness Measurement of Lyophilized Samples
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After lyophilization, a thickness test was performed using the Mitutoyo Dial Thickness Gauge, which provided an accuracy of 0.001 mm. Thickness was measured at five different positions (one in the center and four in the middle of each side).
10
Mouse Model of Atopic Dermatitis
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The mouse AD-like skin lesions model was induced according to the method described in our previous report (25 (link)). A schematic diagram of the experiment is shown in Figure 1A
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