The mice were fasted for 5 h and then intraperitoneally injected with insulin aspart (1 U/kg; Novo Nordisk A/S, Bagsvaerd, Denmark). Blood samples were collected from the caudal vein at 0, 30, 60, 90, and 120 min after injection. Blood glucose level was measured using a glucose meter (FreeStyle Freedom Lite; NIPRO).
Freestyle freedom lite
Manufactured by Nipro
Sourced in Japan
The FreeStyle Freedom Lite is a compact and portable blood glucose monitoring system. It is designed to measure blood glucose levels quickly and conveniently. The device features a simple and intuitive interface, allowing users to easily track and manage their glucose readings.
Automatically generated - may contain errors
8 protocols using freestyle freedom lite
1
Insulin-Induced Glucose Dynamics
2
Measuring Stress Hormone Levels
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An indwelling needle was inserted into a median cubital vein after breakfast and a 12-mL venous blood sample was taken at each time point. The glucose concentrations were measured by using an automated glucose analyzer (FreeStyle FreedomLite; Nipro, Osaka, Japan). The blood samples were each dispensed into a collection vessel for the analysis of adrenocorticotropic hormone (ACTH), epinephrine, and norepinephrine (NE). All samples were centrifuged at 3000 rpm for 15 min at 4°C. The levels of plasma ACTH, epinephrine, and NE were analyzed by means of reverse-phase isocratic high-performance liquid chromatography (HPLC) at a clinical laboratory (MEDIC, Yasu, Japan).
3
Stem Cell Transplant for Diabetic Mice
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Hyperglycemia was induced in adult male C.B-17/IcrHsdPrkdcscid mice via an
intraperitoneal injection of 50 mg/kg of streptozotocin (STZ, Sigma-Aldrich) for five
consecutive days. Blood glucose levels were determined using a blood glucose meter
(Freestyle Freedom Lite; NIPRO, Osaka, Japan). Mice with blood glucose levels >300 mg/dl
were determined as diabetic . Mice were randomly assigned to four study groups:
Venus-hASC, PDX1-2A-Venus-hASC, PM2A-Venus-hASC, and PN-2A-Venus-hASC transplanted (n=15).
Under general anesthesia, mice received a renal subcapsular transplant of
2.0×105 cells in the right subcapsular renal space. Blood glucose levels were
monitored every two days after transplantation for five weeks. All experimental procedures
were approved by the Kindai University Animal Care and Use Committee and were carried out
according to institutional animal experimentation regulations.
intraperitoneal injection of 50 mg/kg of streptozotocin (STZ, Sigma-Aldrich) for five
consecutive days. Blood glucose levels were determined using a blood glucose meter
(Freestyle Freedom Lite; NIPRO, Osaka, Japan). Mice with blood glucose levels >300 mg/dl
were determined as diabetic . Mice were randomly assigned to four study groups:
Venus-hASC, PDX1-2A-Venus-hASC, PM2A-Venus-hASC, and PN-2A-Venus-hASC transplanted (n=15).
Under general anesthesia, mice received a renal subcapsular transplant of
2.0×105 cells in the right subcapsular renal space. Blood glucose levels were
monitored every two days after transplantation for five weeks. All experimental procedures
were approved by the Kindai University Animal Care and Use Committee and were carried out
according to institutional animal experimentation regulations.
4
Oral Glucose Tolerance Test in Rats
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During the 8th week of feeding, the animals were fasted for 18 h preceding an oral Glu tolerance test (OGTT). A sample of blood was taken from the caudal vein for the determination of the pre-load (control) Glu concentration, and then, the animals were given 3 g/kg of Glu by gavage. Thereafter, blood was taken every 30 min up to 150 min after Glu loading. One drop of each blood sample was used for determination of plasma Glu concentration using a coulometric detector for oxidization of Glu to gluconolactone, using a FreeStyle Freedom Lite™ (Nipro, Osaka). The rest of the blood was left for 30 min at room temperature, and then, it was centrifuged at 1200 × g for 30 min. The sera obtained were used for determination of insulin concentration using an ELISA kit (Ultra Sensitive Rat Insulin kit™, Morinaga Institute of Biological Science, Yokohama, Japan), following the instructions of the kit.
5
Measuring Metabolic and Inflammatory Markers in Mice
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Blood sugar levels were measured using FreeStyle Freedom Lite (NIPRO Corporation, Osaka,
Japan). Plasma insulin concentration was measured using a Mouse Insulin ELISA KIT
(Shibayagi Co., Ltd., Shibukawa, Japan). The concentration of macrophage inflammatory
protein 2 (MIP-2) in plasma was measured using a Mouse CXCL2/MIP-2 Quantikine ELISA Kit
(R&D Systems, Inc., Minneapolis, MN, USA). The lipid peroxidation products,
10-hydroxy-8E,12Z-octadecadienoic acid
(10-(Z,E)-HODE) and
12-hydroxy-9Z,13E-octadecadienoic acid
(12-(Z,E)-HODE), were obtained from Larodan Fine Chemicals (Solna,
Stockholm, Sweden). The levels of HODEs were measured by the slightly modified method of a
previously report [38 (link)].
Japan). Plasma insulin concentration was measured using a Mouse Insulin ELISA KIT
(Shibayagi Co., Ltd., Shibukawa, Japan). The concentration of macrophage inflammatory
protein 2 (MIP-2) in plasma was measured using a Mouse CXCL2/MIP-2 Quantikine ELISA Kit
(R&D Systems, Inc., Minneapolis, MN, USA). The lipid peroxidation products,
10-hydroxy-8E,12Z-octadecadienoic acid
(10-(Z,E)-HODE) and
12-hydroxy-9Z,13E-octadecadienoic acid
(12-(Z,E)-HODE), were obtained from Larodan Fine Chemicals (Solna,
Stockholm, Sweden). The levels of HODEs were measured by the slightly modified method of a
previously report [38 (link)].
6
Fasting Metabolism Hormone Analyses in Mice
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Blood samples were collected from the caudal vein of mice that were fasted for 16 h or fed randomly. Blood glucose was measured using a glucose meter (FreeStyle Freedom Lite, NIPRO, Osaka, Japan). Blood samples were centrifuged at 3,000 rpm for 10 min at 4°C to obtain serum, and fasting serum insulin level was measured using a commercial enzyme-linked immunoassay (ELISA) kit (Morinaga Institute of Biological Science, Inc., Yokohama, Japan). Homeostatic model assessment for insulin resistance (HOMA-IR) was performed using the following formula: fasting blood glucose (mg/dL) × fasting immunoreactive insulin (µU/mL)/405.
Blood samples were collected from the inferior vena cava of euthanized mice for metabolic hormone measurement, and serum samples were prepared as described earlier. Serum leptin, fibroblast growth factor 21 (FGF21), and adiponectin were measured using commercial ELISA kits (for leptin, Morinaga Institute of Biological Science, Inc.; for FGF21 and adiponectin, R&D Systems, Inc., Minneapolis, MN, United States).
Blood samples were collected from the inferior vena cava of euthanized mice for metabolic hormone measurement, and serum samples were prepared as described earlier. Serum leptin, fibroblast growth factor 21 (FGF21), and adiponectin were measured using commercial ELISA kits (for leptin, Morinaga Institute of Biological Science, Inc.; for FGF21 and adiponectin, R&D Systems, Inc., Minneapolis, MN, United States).
7
Comparative Pathogenicity of SDSE and GAS
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SDSE-167 was intraperitoneally (i.p.) injected into mice at the indicated concentrations. To compare the pathogenicity of SDSE and GAS, we also injected mice with GAS-M1-476 (also named M1-d). This strain was isolated from a patient with STSS and has the highest lethality for mice of the agents in our collection (Miyoshi-Akiyama et al., 2003 (link), 2012 (link)). These mice were euthanized using sevoflurane (Maruishi Pharmaceutical CO., Ltd.), followed by collection of blood and organs. Glucose concentrations of blood collected from tail veins were measured using NIPRO FreeStyle Freedom Lite (NIPRO, Japan).
8
Blood Metabolite Measurement Protocol
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Blood was obtained from the mice, and serum glucose was measured with a glucometer (FreeStyle Freedom Lite, NIPRO, Osaka, Japan). Serum levels of NEFA, TG and total cholesterol (TC) were measured by enzymatic colorimetry with NEFA C [34 (link)], TG E [35 (link)] and TC E [36 (link)] test kits (Wako Pure Chemical Industries, Ltd., Osaka, Japan), respectively.
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