The lipid peroxidation in Chlamydomonas cells were induced using heat stress. The cell were exposed solely to heat stress and any effect of light was prevented. Samples were treated for 10, 20 and 30 min at temperature of 40 °C using in a water bath (Julabo GmbH, Germany) in Eppendorf tubes.
Water bath
Manufactured by Julabo
Sourced in Germany, Italy, United Kingdom
The Julabo Water Bath is a laboratory equipment designed to maintain a consistent temperature in a water-filled container. It provides a controlled environment for various applications that require temperature regulation, such as sample incubation, reagent preparation, and other temperature-dependent processes.
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Lab products found in correlation
Multichannel peristaltic pump, by Cole-Parmer (3 mentions)
ζsizer nano series zs90, by Malvern Panalytical (3 mentions)
Lse 5003, by Julabo (3 mentions)
Lambda bio 20 spectrophotometer, by PerkinElmer (2 mentions)
Jds uniphase 22 mw he ne laser, by Julabo (2 mentions)
Uniphase 22 mw he ne laser, by Julabo (2 mentions)
Analytical balance, by Sartorius (1 mentions)
Rotary evaporator, by Büchi (1 mentions)
Tween 80, by Merck Group (1 mentions)
Span 80, by Merck Group (1 mentions)
Stereomicroscope, by Thermo Fisher Scientific (1 mentions)
Glass vial, by Avantor (1 mentions)
Dx 120, by Thermo Fisher Scientific (1 mentions)
Alv 60x0 software v 3 x, by Malvern Panalytical (1 mentions)
Zsizer nano series zs90, by Malvern Panalytical (1 mentions)
Nylon syringe filter, by Agilent Technologies (1 mentions)
Agilent 1100 hplc system, by Agilent Technologies (1 mentions)
Zorbax nh2 column, by Agilent Technologies (1 mentions)
25 protocols using water bath
1
Lipid Peroxidation in Chlamydomonas under Heat Stress
2
Reducing Capacity of Plant Extracts
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To check reducing capacity different concentrations of the plant extracts (100–1000 μg mL-1) were mixed with 1 mL phosphate buffer (0.2 M, pH 6.6). Thereafter 1 mL of K3Fe(CN)6 (10 mg mL-1) was added to the reaction and incubated at 50°C in water bath (Julabo, Germany). After 20 min of incubation, 1 mL trichloroacetic acid (100 mg L-1) was added to terminate the reaction. Reaction mixtures were cooled at room temperature, centrifuged at 7000 x g for 10 min and the supernatant was collected. In the next step 1 mL supernatant was mixed with 0.2 mL freshly prepared FeCl3 (0.1% w/v), incubated for 10 min at room temperature, absorbance was measured at 700 nm. Ascorbic acid was used as standard [49 (link), 51 ].
3
Hydroxyl Radical Scavenging Assay for Plant Extracts
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Hydroxyl radical scavenging activity was performed with different concentrations (10–100 μg) of plant extracts using Fenton reaction (Fe3+-ascorbate-EDTA-H2O2) as described by Saeed et al. [54 ]. Plant extracts were mixed with 500 μL of 2.8 mM 2-deoxyribose prepared in 50 mM potassium phosphate buffer (pH 7.4). Thereafter, 200 μL of 100mM FeCl3 and 100mM EDTA solution (1:1 v/v) and 100 μL of 200 mM H2O2 were added to reaction mixture. Reaction was started by adding 100 μL of 300mM ascorbic acid to the reaction mixture and incubated for 1 h at 37°C. After incubation, 500 μL reaction mixture was added to the 1 mL of TCA solution (2.8% w/v) followed by addition of 1 mL of aqueous TBA solution (1% prepared in 0.025 M NaOH containing 0.02% BHA) and incubated at 99°C in water bath (Julabo, Germany) for 15 min. Reactions were cooled at room temperature and absorbance was recorded at 532 nm. The following formula was used to calculate percent scavenging activity.
4
Bronchorelaxant Effects of Waltheria indica
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The apparatus used to achieve the bronchorelaxant effect of extracts from leafy stems of Waltheria indica includes the following: an isolated organ study device equipped with four organ bath chambers (of 20 mL each), isometric transducers (Emka Technologies, France) for measuring isometric force, an amplifier (Emka Technologies, France) for recording changes in isometric force, and a computer for data acquisition. Other instruments used include water bath (Julabo, GmbH, Germany), analytical balance (Sartorius, Germany), oven (Memmert GmbH, Germany), and rotary evaporator (Buchi, Switzerland).
5
Synthesis and Optimization of APM-Functionalized Gold Nanoparticles
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To obtain APM-AuNPs, 0.4 ml of APM aqueous solution (5 mg/ml) and 200 μl of NaOH (100 mM) were added to 9.3 ml of deionized water and incubated in a water bath (Julabo, Seelbach/Black Forest, Germany) (60°C, 200 rpm) for 10 min. Then, 100 μl of HAuCl4 (100 mM) was added followed by vigorous stirring for 30 s. Finally, the mixture was incubated for an additional 50 min (60°C, 200 rpm). The effects of several parameters, such as APM concentration (3.4 × 10−6 M to 1.7 × 10−4 M), HAuCl4 concentration (1.0 × 10−4 M to 9.0 × 10−4 M), pH (2.0 to 12.0), and temperature (25°C/37°C/60°C/90°C) were systematically investigated.
6
Stability Assessment of Lipid Nanoparticles
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To evaluate GSE-ads-DA-SLNs and DA-co-GSE-SLNs physical stability, measurements of their particle dimensions were carried out, storing at 4 °C in the refrigerator up to 3 months, as well as at 25 °C and 37 °C for up to one week and 24 h, respectively. Precisely, a water bath (Julabo, Milan, Italy), set at the temperatures of 25 °C and 37 °C, with agitation of 40 rpm/min, allowed SLN incubation for the above-mentioned time frames. At different time points, particle size was acquired according to the analysis described in Section 2.4 . For each type of SLN, the assay was performed in triplicate at each temperature.
7
Reducing Power of Seaweed Extracts
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The reducing power of seaweed extracts was calculated by using ascorbic acid as standard. Different concentrations of seaweeds extracts (100–500 μg·mL−1) were mixed with 1 mL of phosphate buffer (0.2 M, pH 6.6) and 1 mL of K3Fe(CN)6 (10 mg·mL−1) and was then incubated for 20 min at 50 °C in water bath (Julabo, Seelbach, Germany). The reaction was ended by addition of 1 mL of TCA (100 mg L−1) in the reaction mixture. Supernatant was collected after centrifugation at 7000× g for 10 min at room temperature. Freshly prepared FeCl3 (0.1%, w/v) was mixed with collected supernatant and incubated for 10 min at room temperature. Absorbance was taken at 700 nm to calculate reducing power [34 (link),43 (link)].
8
SNEDDS Dilution and Temperature Rise
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It was measured after 100-times dilution of SNEDDS (1 g) with purified water and set aside for a regular rise in water bath (Julabo, Germany) temperature at 10°C/minute increment.46 (link)
9
Pineal Gland Superfusion Assay
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The pineal glands were covered with a nylon mesh and placed into the culture chambers (volume 0.5 ml). The lower pool of each chamber was connected via a system of tubes and valves to the containers with culture media. The upper pool of the culture chamber was attached to a multichannel peristaltic pump (Cole Parmer, USA) and a manual fraction collector. The total volume of the superfusion set consisting of culture chamber, tubes and valves was 1.3-1.4 ml. The superfusion was performed at a flow rate of 0.1 ml/min. The medium was continuously gassed with a mixture of 95% O 2 and 5% CO 2 . The incubation was performed at 38.5 o C in a water bath (Julabo, Germany). The culture chambers were covered with translucent plastic sheets during the incubation in light and with similar light-proof sheets during the incubation in darkness. The chambers were illuminated with a full-spectrum fluorescent lamp providing light with intensity of 100 lx at the surface of sheets covering the perfusion chambers. The medium fractions were collected every 30 minutes during consecutive days designated as the day 1, 2, 3, 4, 5 and 6, and frozen at -20 o C until melatonin assay. The time-point 07.00 was taken as the beginning of day 2 and following ones.
10
Mucoadhesion Assessment of GSE/DA-SLN
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The mucoadhesive properties of GSE/DA- SLN were assessed in Simulated Nasal Fluid (SNF) by turbidimetric measurements [65 ]. SNF was prepared after dissolution of CaCl2 2H2O (0.32 mg/mL), KCl (1.29 mg/mL) and NaCl (7.45 mg/mL) in water at pH values in the range 5–6 [66 (link)].
To 6 mL of freshly prepared mucin dispersions in SNF (1 mg/mL) held in a water bath (Julabo, Milan, Italy) at 37 °C under stirring (150 rpm), freeze-dried GSE/DA-SLN, previously dispersed in 6 mL of SNF, were added. The turbidity of the stirred mixture at 37 °C was measured at 0, 2, 5, 7 and 24 h at the wavelength of 650 nm using a Perkin-Elmer Lambda Bio 20 spectrophotometer. HEC dissolved in SNF (0.4 mg/mL) was taken as a positive control. Each experiment was performed in triplicate and the results are expressed as mean ± standard deviation of each mean.
To 6 mL of freshly prepared mucin dispersions in SNF (1 mg/mL) held in a water bath (Julabo, Milan, Italy) at 37 °C under stirring (150 rpm), freeze-dried GSE/DA-SLN, previously dispersed in 6 mL of SNF, were added. The turbidity of the stirred mixture at 37 °C was measured at 0, 2, 5, 7 and 24 h at the wavelength of 650 nm using a Perkin-Elmer Lambda Bio 20 spectrophotometer. HEC dissolved in SNF (0.4 mg/mL) was taken as a positive control. Each experiment was performed in triplicate and the results are expressed as mean ± standard deviation of each mean.
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