Polytron pt 3100

Nutraceutical compounds and antioxidant capacity were analysed in the pulp of eggplant fruit. Samples were peeled, cut into pieces and homogenised (Polytron PT 3100, Kinematica AG.,) at 15,000 g for approximately 1 min. Final extracts were divided into aliquots of 2 g, frozen in liquid nitrogen and stored at −80°C until further determinations were made.

Approximately 1.0 g (0.99–1.01 g) of the STP was accurately weighed out into a 50 ml centrifuge tube (Fisher Scientific). 4 ml of water were added and the mixture was equilibrated for 16 h at room temperature. 10 ml of methanol were added and the mixture was macerated (Polytron PT3100, Kinematica AG) at 10,000 rpm for 1 min. The suspension was sonicated at 40 °C for 10 min and shaken (KS501 Flatbed Shaker, Janke and Kunkel) for 30 min at 100 rpm. After centrifuging at 4600 rpm for 5 min, the supernatant was transferred to a 40 ml amber vial and the remaining solvent was squeezed out using a syringe (Discardit 20 ml, BD) and PTFE filter (GD/XP 25 mm, 0.45 μm; Whatman). A second extraction using 5 ml of methanol was carried out in the same way. The first and second extracts were combined and transferred to a tube labelled “extract” and 5 µl were injected into the HPLC/MS/MS.

Foaming properties for each isolated protein were determined according to the method of Chao and others [29 (link)] with slight modifications. Protein solutions of various concentrations (10, 15, and 20 mg/mL) were prepared with 0.1 M acetate, phosphate, or Tris-HCl buffer solutions, vortexed thoroughly and left to hydrate for 30 min. Samples were homogenized at 20,000 rpm for 1 min using a 20 mm foaming shaft on the Polytron PT 3100 homogenizer (Kinematica AG, Lucerne, Switzerland). The foam was formed in a 50 mL graduated centrifuge tube to determine the volume of foam formed (foaming capacity). The volume of foam remaining after 30 min at room temperature was expressed as a percent value of original foam volume to obtain foam stability.

Gastrocnemius and soleus muscles were collected, homogenized in Trizol solution (Invitrogen) using homogenizer (Kinematica Polytron PT3100) and subjected to total RNA extraction according to the manufacturer’s instructions. Total RNA was treated with RQ1 RNase-free DNase (Promega) and was reverse transcribed into cDNA by SuperScript III Reverse Transcriptase (Invitrogen) according to the manufacturer’s instructions. The cDNA was subjected to quantitative PCR analysis on ABI QuantStudio^® 3 Real-Time PCR System (Applied Biosystems) using SYBR Green Master mix (Thermo Scientific). The thermal cycling protocol was 1 cycle at 50°C for 2 min and 95°C for 10 min, followed by 40 cycles of 95°C for 15 s and 60°C for 1 min. The resultant PCR products were analyzed with ABI QuantStudio Design & Analysis Software v.1.4.3 (Applied Biosystems). The relative levels of gene expression were calculated using the ΔCt method with normalization to GAPDH. DNA sequences of the primer pairs used in this study are listed in Supplementary Table 1.

A high-energy method was used for the formulation of oil-in-water nanoemulsion (O/W NE) [2 (link)]. For this, two phases were prepared. The lipidic phase is composed of different combinations of the cumin, carvi, and coriander essential oils, fitted by RSM. Meanwhile, the aqueous phase contained the binary emulsifier mixture (T80: GA 0.75:0.25 v/v). The emulsification process of O/W NE was performed in two steps at room temperature. Firstly, the primary O/W emulsions were passed by high-speed homogenization using Polytron PT-3100, Kinematica, Switzerland, a rotor-stator homogenizer at 10,000 rpm/5 min. Secondly, nanoscale-O/W emulsions were prepared by high-pressure homogenization (HPH) (NanoVater NV200, Yoshida Kikai, Aichi, Japan) at 100 MPa for 4 cycles [4 (link)]. The post-treatment characteristics of their physico-chemical and biological properties were analyzed, whilst the other techno-functional properties were analyzed only for the optimal NE [4 (link)].

Plant emulsions were made by suspending 5% (w/w) plant protein in reverse osmosis (RO) water and mixing at room temperature for 15 min. Coconut oil was heated to 30 °C and 5% (w/w) was added to the plant protein suspension and pre-homogenised using a rotor-stator homogeniser (Polytron PT3100, Kinematica AG, Malters, Switzerland), prior to homogenisation at 30/300 bar (Panda 2k, GEA NIRO Soavi, Parma, Italy). The pH of the emulsions was set to 6.8 using 10% lactic acid prior to autoclaving at 105 °C for 30 min. Plant emulsions for determining acidification were supplemented with 0.5% (m/v) via a 20% glucose stock and sterile 1% 1.064 mM carboxyfluorescein (Sigma-Aldrich, St. Louis, MO, USA) as described earlier [11 (link)]. To minimise the amount of rich medium transferred into the various plant emulsions during inoculation with bacterial strains, the inoculum prepared in rich medium were diluted with plant suspensions. For this, plant protein suspensions per protein type were created by suspending 5% (w/w) plant protein into RO water and mixing at room temperature for 15 min. The pH of the suspensions was adjusted to 6.8 using 10% lactic acid prior to autoclaving at 105 °C for 30 min. After cooling on ice, the suspensions were supplemented with 0.5% (m/v) via a 20% glucose stock.