D glucose powder

5 × 10⁴ U2OS Bmal1-dLuc cells per well were seeded to an opaque 96-well plate and cultured overnight as described earlier^{46 (link)}. The next day cells were reset by adding dexamethasone (DXM) (0.1 µM final) for 2 h. Then medium was changed to bioluminescence recording media which contains the following in 1L: DMEM powder (sigma D-2902, 10X 1L), 0.35 gr sodium bi-carbonate (tissue culture grade, sigma S5761), 3.5gr D(+) glucose powder (tissue culture grade, sigma G7021), 10 mL 1 M HEPES buffer (Gibco 15,140–122), 2.5 mL Pen/Strep (100 ug/ml), 50 mL 5% FBS and up to 1L sterile milliQ water. Luciferin is added freshly with 0.1 mM final concentration. Molecules were added to the bioluminescence recording media at the desired concentration (0.5% DMSO final concentration). Plates were sealed with optically clear film to prevent evaporation and gas exchange thereby maintaining homeostasis of the cells. Luminescence values were recorded at 32 °C for every 30 min with 15 s integration time via Synergy H1 luminometer for a week. The experiment was repeated three times with 3-technical replicates. To obtain the period values BioDare2 (biodare2.ed.ac.uk) was used^{47 (link)}. Significant analysis was performed by using the unpaired t-test with Welch’s correction.

The glucose-based aqueous solution was prepared to measure the sensing performance of the proposed resonator-based microwave biosensor. The aqueous solution was prepared using a quantized mixture of D-Glucose powder (Sigma Aldrich) and deionized water (Merck Millipore, Billerica, MA). The glucose solution was calibrated using standard concentrations of 0.3, 1, 2, 3, 4, and 5 mg/mL. The glucose sample’s range (0.3–5 mg/mL) was selected to cover diabetes patients with hyperglycemia (glucose concentration > 1.20 mg/mL) as well as hypoglycemia (glucose concentration < 0.80 mg/mL) [50 ]. A one hundred-nanoliter (100 nL) droplet was dropped on the sensing area with a digitally variable pipette (0.1–2.5 μL), purchased from Shanghai LC-BX Instrument Technology Co Ltd., to quantify the sample volume of the glucose solution. All samples are tested at room temperature with a fixed-volume pipette to achieve a fixed volume and shape.

Dermal papilla cells (DP; PromoCell GmbH, Heidelberg, Germany, Lot 325Z017.1 and 322Z030.1) were maintained in the Follicle Dermal Papilla Cell Basal Medium (PromoCell GmbH) supplemented with 4 µl/ml bovine pituitary extract, 0.04 ml/ml fetal calf serum, 1 ng/ml basic recombinant human fibroblast growth factor, 5 µg/ml recombinant human insulin (PromoCell). Cells were cultured with 50 mg/ml primocin (Invivogen, Toulouse, France, #ant-pm-2) in a humidified 5% CO₂ incubator at 37 °C. cDP and sDP were maintained as previously reported^{4 (link)}. Briefly, cDP were maintained in 60 mm dish (SPL, Gyeonggi-do, Korea, #20060) and sDP were maintained in Ultra Low Attachment Culture Dish (Corning, New York, USA #3261). Both cDP and sDP were maintained same DP media described above. 2-deoxy-d-glucose (2DG), WZB117 and Bromopyruvic acid (3BP) were purchased from Sigma–Aldrich (St. Louis, USA, #D8375, #SML0621, #16490 respectively). D-(+)-Glucose powder were purchased from Sigma–Aldrich (#G7021). C646 (#10549) and Ginkgolic acid (#18422) were purchased from Cayman Chemical (Ann Arbor, USA).

Laboratory‐reared An. coluzzii and An. gambiae were obtained from outbred colonies established in 2019, which have since been repeatedly replenished with wild‐caught gravid females collected in the Vallée du Kou (11°23′N, 4°24′W) and Soumousso (11°04′N, 4°03′W), respectively, in southwestern Burkina Faso, and identified by SINE PCR (Santolamazza et al., 2008 (link)). Females were maintained on rabbit blood by direct feeding (protocol approved by the national committee of Burkina Faso; IRB registration #00004738 and FWA 00007038) for egg production. Larvae were reared in 1 L of tap water in plastic trays and fed daily with TetraMin® Baby Fish Food (Tetrawerke, Melle, Germany) until adulthood. The adult mosquitoes were held in 30 cm × 30 cm × 30 cm mesh‐covered cages under standard controlled conditions (27 ± 2°C, 70 ± 5% RH, and at a 12 h:12 h light:dark rhythm), and emerged males and females were fed daily with 5% glucose. This solution was prepared using D‐(+)‐glucose powder, Sigma‐Aldrich®, and distilled water.

Saccharomyces cerevisiae 21PMR (MAT leu2 ura3-52) was used throughout the conducted experiments. D-glucose powder, yeast extract, peptone powder, and carbon nanotubes were obtained from Sigma-Aldrich (Steinheim, Germany). Double distilled water has been used throughout the experimental work. All other essential reagents and chemicals used in the study were of analytical grade.
For biofuel cell, dried yeast was purchased from a food supplier “Dr. Oetker Lietuva” (Vilnius, Lithuania). 1 g of YPD-broth was mixed with 20 mL distilled water to get medium with YPD-broth 50 g/L concentration. 500 mg of dried yeast was introduced to prepared suspension. A further culture was grown in shaking incubator at 200 rpm till yeast reaches Logarithmic Phase (20–24 h).

The high-fat diet used was modified from that described previously [11 (link)]. The ingredients were a mixture of 40% control diet (Smart Heart; PCG, Bangkok, Thailand), 10% coconut oil (Roi Thai, Thailand), 10% corn oil (Golden Drop, Thailand), 1.25% cholesterol, 0.25% sodium deoxycholate (Sigma-Aldrich, St, Louis, Mo, USA), and 38.5% sucrose (Mitr Phol, Thailand). All of the ingredients were combined and formed into small discs, which were then incubated at 65°C for 24 h. The high-fructose drinking water was prepared by mixing d-glucose powder (18.9 g) and d-fructose powder (23.1 g) (Merck Millipore, Darmstadt, Germany) in distilled water (1 L) [30 (link)]. HFF diet composition was determined by Central Laboratory (Thailand), Co. Ltd. according to the Association of Office Analytical Collaboration (AOAC) International guidelines and is shown in S1 Table.