Blood samples collected from the chickens (n = 48 chickens) were used to assay the biochemical parameters. Total protein, albumin, triglycerides (TAG), and glucose were determined biochemically in the collected sera according to the manufacturing instructions of the Biodiagnostic company kits (Dokki, Giza, Egypt; www.bio-diagnostic.com , accessed on 12 March 2020). The total protein and albumin differences in the collected samples were used to determine the globulin levels. The biochemical parameters were measured using a UV-VIS spectrometer (model T60UV, PG Instruments Limited, Lutterworth, UK). Samples were taken from chickens in the high-weight (HW) group and low-weight (LW) group (n = 48 chickens, divided into 12 males and 12 females for each group).
T60uv
Manufactured by PG Instruments
Sourced in United Kingdom
The T60UV is a spectrophotometer designed for absorbance and transmittance measurements in the ultraviolet and visible light ranges. It features a wavelength range of 190 to 1100 nm and can be used for a variety of analytical applications.
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26 protocols using t60uv
1
Biochemical Analysis of Chicken Serum
2
Citrate Synthase Enzyme Activity Assay
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Citrate synthase (CS) enzyme activity was assessed as previously described[30 (link),33 (link)]. From 40 to 70 mg samples of the gastrocnemius muscle (n=8 in each group) were homogenized in ice-cold lysis buffer (50 mM Tris-HCl, 100 mM KHPO4, 2 mM ethylenediaminetetraacetic acid (EDTA), 0.2% wt/vol bovine serum albumin, pH was adjusted to 7.0) in frozen liquid nitrogen. Then, the homogenates were defrosted by shaking for 60 min and centrifuged at 13,000 g for 10 min at 4°C for extraction of soluble proteins. The Bradford reagent (B6916, Merck, Germany) was used for assessment of protein concentration of supernatants. Measurements of CS activity were carried out using CS reaction reagent (100 mM triethanolamine-HCl, 100 µM dithionitrobenzoic acid (DTNB), 0.5 mM Triton-X (0.25% vol/vol), 0.5 mM oxaloacetate, 0.31 mM acetyl CoA, pH 8.0) and spectrophotometer (T60 UV, PG Instruments Limited, UK) at room temperature of 21°C. 10 µL of supernatant was added to start the reaction in 1000 µL. The CS from a porcine heart was used as a standard (C3260, Merck, Germany) for assay calibration. The wavelength of 412 nm and molar extinction coefficient of 13,600 M-1 cm-1 were used to assess the maximum CS activity (Vmax) during the first 2 min of the reaction.
3
Determination of Total Flavonoids in Sacha Inchi
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Total flavonoid contents in Sacha inchi were determined using the aluminum chloride colorimetric method described by Woisky and Salatino (1998) [8 (link)]. The appropriate dilution of extractions (0.5 mL) was briefly mixed with 1.5 mL of 95% ethanol, 0.1 mL of 10% aluminum chloride hexahydrate, 0.1 mL of 1 M potassium acetate, and 2.8 mL of deionized water. After incubation at room temperature for 40 min, the absorbance of the reaction mixture was measured at 415 nm against a deionized water blank on a UV visible spectrophotometer (T60UV, PG instruments Ltd., Lutterworth, UK).
4
Determination of Total Polyphenols
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Polyphenolic compounds (1 mM) were prepared in ethanol; 40 μL of Sacha inchi extract was transferred into a 10 mL screw-cap tube. Then, 800 μL of a 10-fold diluted folin ciocalteau reagent was added into the tube and mixed well. The tube was allowed to stand for 5 min. Then, 800 μL of 7% sodium carbonate aqueous solution (w/v) was added to the tube and mixed well. The volume in the tube was made up with nano pure water (360 μL), mixed well, and then allowed to stand for 2 h at room temperature. Absorbance was read at 760 nm against the blank using a UV visible spectrophotometer (T60UV, PG instruments Ltd., Lutterworth, UK).
5
Evaluating Biochemical Changes in Aged Foods
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For pH, reducing sugars and browning intensity analysis, 10 g of the freeze-dried powder samples according to the aging period was added to 10-fold distilled water, homogenized for 3 min at 1500 rpm with a homogenizer (PT-2100, Kinematica Ag, Littau, Switzerland) and then the supernatant was filtered. The pH was measured using a pH meter (PB 30, Sartorius, Goettingen, Germany), and the browning intensity was measured with a UV–Vis spectrophotometer (T60UV, PG Instruments, Wibtoft, UK) at an absorbance of 420 nm.
The determination of reducing sugar content was carried out using the DNS method [20 (link)]. The freeze-dried powder sample according to the aging period (1 mL) was mixed with 3 mL of the DNS reagent and shaken slightly. The mixture solution was heated in a 75 °C water bath (WBT-10, Jeongbiotech co., Incheon, Republic of Korea) for 5 min. After heating, the mixture was cooled in running water for 10 min. The absorbance at 575 nm was determined. The standard curve used glucose, and the reducing sugar content of the EG extract according to the aging period was expressed as the equivalent of D-glucose per 1 mL of the sample.
The determination of reducing sugar content was carried out using the DNS method [20 (link)]. The freeze-dried powder sample according to the aging period (1 mL) was mixed with 3 mL of the DNS reagent and shaken slightly. The mixture solution was heated in a 75 °C water bath (WBT-10, Jeongbiotech co., Incheon, Republic of Korea) for 5 min. After heating, the mixture was cooled in running water for 10 min. The absorbance at 575 nm was determined. The standard curve used glucose, and the reducing sugar content of the EG extract according to the aging period was expressed as the equivalent of D-glucose per 1 mL of the sample.
6
Folin-Ciocalteu Assay for Total Phenolic Content
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TPC was determined based on the Folin–Ciocalteu method by Hillis and Swain [21 (link)]. Briefly, 150 μL of freeze-dried extract, 2400 μL of distilled water and 50 μL 2 N Folin–Ciocalteu reagent mixtures were reacted at room temperature for 3 min. Then, after 300 µL of 5% Na2CO3 was added, the mixtures were reacted in the dark for 2 h. The absorbance was measured using a UV/Vis spectrophotometer (T60UV, PG Instruments, Wibtoft, England) at 725 nm. Gallic acid was used as the standard compound, and TPC results were expressed as milligram gallic acid equivalents (GAE) per gram of extract.
7
Photosynthetic Traits and Leaf Pigments
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Gas exchange features, including photosynthesis rate (Pn), stomatal conductance to water (H 2 O) (g s ) and transpiration rate (E) of mung bean plants were measured using a LI-6400XT portable photosynthesis system (LI-COR Biosciences, Lincoln, Nebraska, USA) from 11.00 AM to 2.00 PM under a full sun-light condition. The parameters Pn, g s and E were used for assessing the instantaneous water-use e ciency (WUEins; ratio Pn/E) and intrinsic water-use e ciency (WUEint; ratio Pn/g s ).
2.4. Quanti cation of photosynthetic pigments, and total phenolic and avonoid contents in mung bean leaves
The freshly harvested mung bean leaves were used to determine the contents of chlorophylls (Chls) [Chl a, Chl b and Chl (a + b)] by using the visible spectrophotometer (Model: T60 UV, PG Instruments Limited, Leicestershire, UK) according to the protocols described by Lichtenthaler and Wellburn (1983) . The contents of total phenolic and total avonoid were quanti ed following the methods of Ainsworth and Gillespie (2007) , and Zhishen et al. (1999) , respectively.
2.4. Quanti cation of photosynthetic pigments, and total phenolic and avonoid contents in mung bean leaves
The freshly harvested mung bean leaves were used to determine the contents of chlorophylls (Chls) [Chl a, Chl b and Chl (a + b)] by using the visible spectrophotometer (Model: T60 UV, PG Instruments Limited, Leicestershire, UK) according to the protocols described by Lichtenthaler and Wellburn (1983) . The contents of total phenolic and total avonoid were quanti ed following the methods of Ainsworth and Gillespie (2007) , and Zhishen et al. (1999) , respectively.
8
Determination of Chlorophyll and Carotenoid
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Determination of the chlorophyll and carotenoid contents was performed following Arnon (1949) (link) method. Freshly grown shoots, weighing 5 g, were ground in 10 ml acetone (80%). The filtrate was collected and the absorbance of the filtrate determined at 480, 645, and 663 nm using a spectrophotometer (model T60UV; PG Instruments Limited, Lutterworth, UK).
9
Antioxidant Evaluation of Co-ZnO Nanoparticles
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Antioxidant activity of Co‐ZnO NPs was determined using 2,2 diphenyl‐1 picrylhydrazil (DPPH). The method was modified
39 (link) and ascorbic acid was used as the standard. 1000 μL of DPPH solution in methanol (0.1 mM) was added to the samples containing 1000 μL of Co‐ZnO NP. After adding DPPH to the samples, they were incubated for 30 min at room temperature and in the dark, and their absorbance was measured at 517 nm (T60 UV‐PG Instruments). As a negative control, methanol was used instead of the sample. The % inhibition is calculated using the following formula: where Acontrol is the absorbance of negative control and Asample is the absorbance of sample.
39 (link) and ascorbic acid was used as the standard. 1000 μL of DPPH solution in methanol (0.1 mM) was added to the samples containing 1000 μL of Co‐ZnO NP. After adding DPPH to the samples, they were incubated for 30 min at room temperature and in the dark, and their absorbance was measured at 517 nm (T60 UV‐PG Instruments). As a negative control, methanol was used instead of the sample. The % inhibition is calculated using the following formula: where Acontrol is the absorbance of negative control and Asample is the absorbance of sample.
10
Biochemical Profiling of Rabbit Breeds
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Before euthanasia, blood samples were collected from all the rabbits (n = 10 for each breed) and centrifuged at 1500× g for 20 min; the serum samples were then kept at −80 °C for further biochemical parameter analyses: glucose, total protein, albumin, globulin, triglycerides (TG), and urea were analyzed according to the manufacturing instructions of the Biodiagnostic company kits (Dokki, Giza, Egypt; www.bio-diagnostic.com , accessed on 22 March 2021). The total protein and albumin differences in the collected samples were used to determine the globulin levels. The biochemical parameters were measured using a UV-VIS spectrometer (model T60UV, PG Instruments Limited, Lutterworth, UK).
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