Scientists present wireless glucose monitoring methodology, best practices, research findings, and discuss the power of continuous glucose profiles, particularly when combined with other gold standard assays to study obesity, diabetes and cardiovascular related diseases. During this webinar sponsored by Data Sciences International (DSI), Stephanie Simonds, PhD shares novel research conducted using implantable glucose telemetry for basic analysis and combination studies investigating the effects of drugs and hormones in mice. Specifically, she discusses research case studies involving Designer Receptors Exclusively Activated by Designer Drugs (DREADS) and adeno-associated virus neuronal knockdown models, and speaks to the importance of collecting continuous glucose data simultaneously with physiologic telemetry measurements. In addition, Dr. Megan Fine from DSI’s Surgical Services team presents the surgical techniques and best practices that her team has developed for use with glucose telemetry. She discusses the use of this technology in a variety of animal models, including recommendations for surgery and techniques to help ensure successful calibrations. Finally, Megan highlights the concurrent collection of multiple endpoints, such as glucose and cardiovascular data, in the same subject.

1. From Mouse to Monkey:
Revolutionizing Research via Preclinical
Continuous Glucose Telemetry
Scientists present wireless glucose monitoring methodology, best practices,
research findings, and discuss the power of continuous glucose profiles,
particularly when combined with other gold standard assays to study obesity,
diabetes and cardiovascular related diseases.

2. From Mouse to Monkey:
Revolutionizing Research via Preclinical
Continuous Glucose Telemetry
Dr. Stephanie Simonds
NHMRC/NHF Research fellow
Monash University,
Melbourne, Australia
Megan Fine, DVM
MS, DACVS-LA
Veterinary Surgeon,
Attending Veterinarian
Data Sciences International

3. InsideScientific is an online educational environment designed for life science
researchers. Our goal is to aid in the sharing and distribution of scientific
information regarding innovative technologies, protocols, research tools and
laboratory services

4. To access webinar content, Q&A reports, FAQ documents, and
information on lab workshops, subscribe to our mail list

5. Copyright 2018 S. Simonds and InsideScientific. All Rights Reserved.
Experimental applications of DSI
continuous blood glucose monitoring
in mice.
Dr. Stephanie Simonds
NHMRC/NHF Research fellow
Monash University, Melbourne, Australia

6. We have used DSI equipment for >9 years
We use this equipment for the study of chronic diseases including
diabetes and cardiovascular diseases
HD-X11
HD-XG
PA-C10

7. Cardiovascular diseases Diabetes
Obesity

8. Lean DIO
0
10
20
30
40
50
60
70
Bodyweight(grams)
***
High Fat Diet-fed Mouse Model Of Obesity

9. Using Continuous Glucose
Monitoring to Assess Phenotypes

10. Lean DIO
0
10
20
30
40
50
60
70
Bodyweight(grams)
***
Continuous Telemetric Blood Glucose
7:00:009:00:0011:00:0013:00:0015:00:0017:00:0019:00:0021:00:0023:00:001:00:003:00:005:00:00
2
4
6
8
10
Time (Hrs:Min:Sec)
Bloodglucose(mmol/l)
DIO fed
Lean fed
Light Phase Dark Phase
7:00:009:00:0011:00:0013:00:0015:00:0017:00:0019:00:0021:00:0023:00:001:00:003:00:005:00:00
2
Time (Hrs:Min:Sec)
Blo
DIO fed
Lean fed

11. 0:00:004:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
32
34
36
38
Temp(Degrees)
temp when changed to HFD
Switch to HFD
0:00:004:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
0
50
100
150
Activity
(Arbitraryunit)
activity when changed to HFD
0:00:004:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
0
2
4
6
8
10
12
14
Blood
glucose(mmol/l)
BG when changed to HFD
00:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
12:00:00
16:00:00
20:00:000:00:004:00:008:00:00
Switch to HFD
Initial Physiological Changes When Mice are Switched to a HFD

12. Using Continuous Glucose Monitoring
to Assess Drug Therapy Efficacy

13. 0:00:00
0:30:00
1:00:00
1:30:00
2:00:00
2:30:00
3:00:00
3:30:00
4:00:00
4:30:00
5:00:00
5:30:00
6:00:00
6:30:00
7:00:00
0
10
20
30
40
Time (Hrs:Min: Sec)
Blood
glucose(mmol/l)
Drug treatment
Glucose Bolus
Veh
-60 0 30
0
5
10
15
20
25
Time (Mins)
Blood
glucose(mmol/l)
Manual gluose
Veh
Synthetic compound ‘B’
Synthetic compound ‘A’
Native Peptide ‘A’
Combination Drug ‘A’ + ‘B’
-60 0 30
0
5
10
15
20
25
Time (Mins)
Blood
glucose(mmol/l)
Manual gluose
Veh
Synthetic compound ‘B’
Synthetic compound ‘C’
Synthetic compound ‘A’
Native Peptide ‘A’
Synthetic compound ‘D’
Combination Drug ‘A’ + ‘B’
Manual vs. Continuous Monitoring
0 30
Time (Mins)
anual gluose
Veh
Synthetic compound ‘B’
Synthetic compound ‘C’
Synthetic compound ‘A’
Native Peptide ‘A’
Synthetic compound ‘D’
Combination Drug ‘A’ + ‘B’
Manual
Continuous
0:00:00
0:30:00
1:00:00
1:30:00
2:00:00
2:30:00
3:00:00
3:30:00
4:00:00
4:30:00
5:00:00
5:30:00
6:00:00
6:30:00
7
0
Time (Hrs:Min: Sec)
Synthetic compound ‘C’
Synthetic compound ‘A’
Native Peptide ‘A’
Synthetic compound ‘D’
Synthetic compound ‘E’
Combination Drug ‘A’ + ‘B’
Synthetic compound ‘B’
Veh
0:00:00
0:30:00
1:00:00
1:30:00
2:00:00
2:30:00
3:00:00
3:30:00
4:00
Time (Hrs:Min:
Synthetic compound ‘C’
Synthetic compound ‘A’
Native Peptide ‘A’
Synthetic compound ‘D’
Synthetic compound ‘E’
Combination Drug ‘A’ + ‘B’
Synthetic compound ‘B’
Veh
Veh
Compound ‘F’
Compound ‘A’
Compound ‘B’
Compound ‘A’ + ‘B’
Compound ‘C’
Compound ‘D’
Veh
Compound ‘F’
Compound ‘A’
Compound ‘B’
Compound ‘A’ + ‘B’
Compound ‘C’
Compound ‘D’
Compound ‘E’

14. Using Continuous Glucose Monitoring to
Determine Hormone Physiological Actions

15. https://www.ncbi.nlm.nih.gov/pubmed/8603777
More than 20 years ago…
Zymogenetics Corporation is a wholly owned subsidiary of Novo
Nordisk A/S, a manufacturer of therapeutics for the treatment of
diabetes.
BBB, blood-brain barrier; CRH, corticotropin-releasing hormone;
MES, 2-[Nmorpholino]ethanesulfonic acid; NPY, neuropeptide Y;
PVN, paraventricular nucleus.
DIABETES, VOL. 45, APRIL 1996
(A) Serum glucose in ob/ob and db/db mice after 5 daily intraperitoneal
injections of 150 µg of recombinant murine leptin or saline
genotype: ob/ob db/db
treatment:
Specificity of Leptin Action on
Elevated Blood Glucose Levels and
Hypothalamic Neuropeptide Y Gene
Expression in ob/ob Mice
Michael W. Schwartz, Denis G. Baskin, Thomas R. Bukowski,
Joseph L. Kuijper, Donald Foster, Gerry Lasser, Donna E.
Prunkard, Daniel Porte, Jr., Stephen C. Woods, Randy J. Seeley,
and David S. Weigle

16. 7:00:0011:00:0015:00:0019:00:0023:00:003:00:007:00:0011:00:0015:00:0019:00:0023:00:003:00:007:00:00
-5.0
-2.5
0.0
2.5
5.0
Time (Hrs:Min:Sec)
Changeinbloodglucosefrombaseline(mmol/l)
Leptin
Saline
Leptin (2ug/g)
*IP leptin
Leptin Acutely Decreases BG

17. Using Continuous Glucose Monitoring
in Combination with Chemogenetics

18. • X-Cre mice
• Injected into the DMH with chemogenic
activator virus
• Mice allowed to recover
• DSI equipment utilized in monitoring of blood
glucose, core body temperature and activity
Study Overview

19. 0:10:000:30:000:50:001:10:001:30:001:50:002:10:002:30:002:50:003:10:003:30:00
0
2
4
6
8
10
Time (Hrs:Min: Sec)
Blood
glucose(mmol/l)
DMH lepR activation
DMH lepR no activation
* ** ** **** ** ****
Either activation or
no activation
0:10:000:30:000:50:001:10:001:30:001:50:002:10:002:30:002:50:003:10:003:30:00
30
32
34
36
38
Time (Hrs:Min: Sec)
Coretemp(Degrees)
DMH lepR activation
DMH lepR no activation
* *** **
Either activation or
no activation
0:10:000:30:000:50:001:10:001:30:001:50:002:10:002:30:002:50:003:10:003:30:00
0
5
10
15
Time (Hrs:Min: Sec)
Activity
(Arbitraryunit)
DMH lepR activation
DMH lepR no activation
*
Either activation or
no activation
DMH receptor activation
DMH veh non- activation
DMH receptor activation
DMH veh non- activation
DMH receptor activation
DMH veh non- activation
Activation vs. Non Activation of X Receptor Expressing Neurons
Within The Dorsomedial Hypothalamic Nucleus

20. • X-Cre mice
• Injected into the DMH with chemogenic activator virus
• Mice allowed to recover
• DSI equipment utilized in monitoring of blood glucose, core
body temperature and activity in response to chemogenic
neuron activation of receptors and in response to blood
glucose bolus
Study Overview

21. 00:10:000:20:000:30:000:40:000:50:001:00:001:10:001:20:001:30:001:40:001:50:002:00:002:10:002:20:002:30:002:40:002:50:003:00:003:10:003:20:00
0
5
10
15
Activity
(Arbitraryunit)
DMH lepR activation + glucose
*** *** *** **
DMH lepR no activation + glucose
***
Activation or
no activation
All mice
Glucose bolus
00:10:000:20:000:30:000:40:000:50:001:00:001:10:001:20:001:30:001:40:001:50:002:00:002:10:002:20:002:30:002:40:002:50:003:00:003:10:003:20:00
0
10
20
30
Blood
glucose(mmol/l)
DMH lepR activation + glucose
***
***
**
*
**
**
** ** ** ** **
DMH lepR no activation + glucose
Activation or
no activation
All mice
Glucose bolus
00:10:000:20:000:30:000:40:000:50:001:00:001:10:001:20:001:30:001:40:001:50:002:00:002:10:002:20:002:30:002:40:002:50:003:00:003:10:003:20:00
30
32
34
36
38
Time (Hrs:Min: Sec)
Coretemp(Degrees)
DMH lepR activation + glucose
DMH lepR no activation + glucose
Activation or
no activation
All mice
Glucose bolus
DMH receptor activation + glucose
DMH veh non- activation + glucose
DMH receptor activation + glucose
DMH veh non- activation + glucose
DMH receptor activation + glucose
DMH veh non- activation + glucose
Physiological Effects of Glucose Bolus and Activation vs. Non Activation of X-cre
Expressing Neurons Within The Dorsomedial Hypothalamic Nucleus

22. Using Continuous Glucose Monitoring in Combination
With Central AAV Cre Neuronal Knockdown
Modulating central physiology & measuring peripheral outputs

23. baseline
15
30
45
60
90
1200
10
20
30
40
Blood
glucose(mmol/l)
Time (Mins)
Baseline
Week 1
Week 3
** ****
X
lox/lox
mice
0:00:000:15:000:30:000:45:001:00:001:15:001:30:001:45:002:00:002:15:002:30:002:45:00
0
10
20
30
40
Time (Hrs:Min: Sec)
Blood
glucose(mmol/l)
DMH AAV Cre in LepR flox mice day 17
DMH AAV Cre in LepR flox mice day 4
Deletion of X-receptor in DMH Impairs Glucose Tolerance
DMH AAV Cre in x Cre receptor mice day 17
DMH AAV Cre in x Cre receptor mice day 4
0:00:000:15:000:30:000:45:001:00:001:15:001:30:001:45:002:00:002:15:002:30:002:45:00
0
10
20
30
40
Time (Hrs:Min: Sec)
Blood
glucose(mmol/l)
DMH AAV Cre in LepR flox mice day 17
DMH AAV Cre in LepR flox mice day 4

24. 19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
-10
-5
0
5
10
Changeinbloodglucose(mmol/l)
change
DMH AAV Cre in LepR flox mice
DMH AAV Scrambled in LepR flox mice
Day 7 Day 14 Day 18
Central AAV-Cre Receptor Deletion Increases Blood Glucose
X
lox/lox
mice
DMH AAV Cre in x Cre receptor mice
DMH AAV Scrambled in x Cre receptor mice
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
19:00:00
7:00:00
-10
-5
0
5
10
Changeinbloodglucose(mmol/l)
change
DMH AAV Cre in LepR flox mice
DMH AAV Scrambled in LepR flox mice
Day 7 Day 14 Day 18

25. 19:00:00
23:00:00
3:00:00
7:00:00
11:00:00
15:00:00
-2
0
2
4
6
8
10
Time (Hrs:Min: Sec)
Changeinbloodglucose(mmol/l)
Change DAY 7
DMH AAV Cre in LepR flox mice
DMH AAV Scrambled in LepR flox mice
19:00:00
23:00:00
3:00:00
7:00:00
11:00:00
15:00:00
-2
0
2
4
6
8
10
Time (Hrs:Min: Sec)
Changeinbloodglucose(mmol/l)
Change DAY 14
DMH AAV Cre in LepR flox mice
DMH AAV Scrambled in LepR flox mice
19:00:00
23:00:00
3:00:00
7:00:00
11:00:00
15:00:00
-5
0
5
10
15
Time (Hrs:Min: Sec)
Changeinbloodglucose(mmol/l)
Change DAY 18
DMH AAV Cre in LepR flox mice
DMH AAV Scrambled in LepR flox mice
Time (Hrs:Min:Sec)
x
lox/lox
mice
Deletion of X-Receptors Centrally Increases Blood Glucose
DMH AAV Cre in x Cre receptor mice
DMH AAV Scrambled in x Cre receptor mice
19:00:00
23:00:00
3:00:00
7:00:00
11:00:00
15:00:00
-2
0
Time (Hrs:Min: Sec)
Chang
DMH AAV Cre in LepR flox mice
DMH AAV Scrambled in LepR flox mice
Time (Hrs:Min:Sec) Time (Hrs:Min:Sec)
Change DAY 18Change DAY 14Change DAY 7

26. Monash University
Prof. Michael Cowley
Dr. Jack Pryor
Acknowledgements

27. Glucose Telemetry Surgery
Copyright 2018 Data Sciences International (DSI) and InsideScientific. All Rights Reserved.
Megan Fine, DVM
MS, DACVS-LA
Veterinary Surgeon, Attending Veterinarian DSI

28. © 2018 Data Sciences International
Overview
• Why blood glucose?
• Glucose telemetry surgery in mouse, rat and large animals
• Calibration best practices
• Combining additional endpoints
• Questions

29. © 2018 Data Sciences International
Why blood glucose?

30. © 2018 Data Sciences International
Why blood glucose?
• Decreased handling
• Minimize stress
• Continuous data
• Reduced variability
• ↑ Statistical power
• Effort to ↓ animal numbers
accessiblebiosci.blogspot.com

31. © 2018 Data Sciences International
Glucose Telemetry Surgery
• Arterial insertion
– Sensor tip in freely
flowing blood
• Temperature from
device body
• Sensor hydration
• Surgical recovery
• Calibration

32. © 2018 Data Sciences International
Glucose Telemetry Surgery
HD-XG Size
Implant Body = ~20mm
Catheter = ~65mm
Total = ~85mm

33. © 2018 Data Sciences International
Mouse Glucose Surgery
• Pre-operative planning
– Ensure complete circle of Willis
• Ligate and recover
Click here to learn more about
HD-XG glucose telemetry

34. © 2018 Data Sciences International
• Pre-operative planning
– Ensure complete circle of Willis
• Ligate and recover
– Insertion depth assessment
• Ligate and measure
Bifurcation
Junction
Mouse Glucose Surgery

35. © 2018 Data Sciences International
• Pre-operative planning
– Ensure complete circle of Willis
• Ligate and recover
– Insertion depth assessment
• Ligate and measure
• Example; 10 mm bifurcation → junction
10 mm
Mouse Glucose Surgery

36. © 2018 Data Sciences International
• Pre-operative planning
– Ensure complete circle of Willis
• Ligate and recover
– Insertion depth assessment
• Ligate and measure
• Example; 10 mm bifurcation → junction
– Determine insertion depths
• Example 12 mm insertion
12 mm
Mouse Glucose Surgery

37. © 2018 Data Sciences International
• Pre-operative planning
– Ensure complete circle of Willis
• Ligate and recover
– Insertion depth assessment
• Ligate and measure
– Determine insertion depths
• Varies by weight, sex & strain
Animal
#
Weight
(g)
Measured
depth (mm)
add
2 mm
1 28.7 11
8 30.3 10 12
9 30.6 12
15 31.4 12.5
21 32.4 13
22 32.4 11 13
29 34.6 14
Mouse Glucose Surgery
Animal # Weight (g)
1 28.7
2 28.8
3 28.9
4 28.9
5 29.4
6 29.5
7 29.8
8 30.3
9 30.6
10 31.1
11 31.3
12 31.3
13 31.4
14 31.4
15 31.4
16 31.6
17 31.8
18 32.1
19 32.1
20 32.2
21 32.4
22 32.4
23 32.5
24 32.6
25 32.7
26 32.9
27 33
28 33.5
29 34.6
30 34.7

38. © 2018 Data Sciences International
Intraperitoneal Placement
• Minimum weight 23 g
• Core body temperature
• Major surgery
Subcutaneous Placement
• Minimum weight 19 g
• Temperature trends
• Care with pocket size
Mouse Glucose Surgery

39. © 2018 Data Sciences International
• Isolate left carotid artery • Isolate left carotid artery
Mouse Glucose Surgery
http://erovideo.us/d1aebe/mouse-vessel-anatomy
Click here to learn more
about DSI surgical services

40. © 2018 Data Sciences International
• Isolate left carotid artery
• Ligate at bifurcation
• Two additional sutures for
hemostasis
Mouse Glucose Surgery
[superior - cranial]
[inferior - caudal]

41. © 2018 Data Sciences International
• Isolate left carotid artery
• Ligate at bifurcation
• Two additional sutures for
hemostasis
• Prep needle for puncture
Mouse Glucose Surgery

42. © 2018 Data Sciences International
• Measure sensor: White marker → tip
12 ± 0.5 mm
Mouse Glucose Surgery

43. © 2018 Data Sciences International
• Grasp sensor
– Vessel cannulation forceps
– On edge
– Behind exposed wells
Mouse Glucose Surgery

44. © 2018 Data Sciences International
• Grasp sensor
– Vessel cannulation forceps
– On edge
– Behind exposed wells
• Puncture and insert
– Remember depth
Mouse Glucose Surgery

45. © 2018 Data Sciences International
• Grasp sensor
– Vessel cannulation forceps
– On edge
– Behind exposed wells
• Puncture and insert
– Remember depth
• Secure with three sutures
• Flat under skin
Mouse Glucose Surgery

46. © 2018 Data Sciences International
Intraperitoneal Placement
• Prior to cannulation
• Midline laparotomy
Subcutaneous Placement
• Tunnel after cannulation
• Right lateral flank
Mouse Glucose Surgery

47. © 2018 Data Sciences International
Rat Glucose Surgery

48. © 2018 Data Sciences International
Rat Glucose Surgery
• Intraperitoneal placement
Rat Glucose Surgery

49. © 2018 Data Sciences International
Rat Glucose Surgery
• Intraperitoneal placement
• Occlusion sutures
– Distal to renal vein
– Proximal to iliac bifurcation
Rat Glucose Surgery

50. © 2018 Data Sciences International
Rat Glucose Surgery
• Intraperitoneal placement
• Occlusion sutures
• Insert sensor
Rat Glucose Surgery

51. © 2018 Data Sciences International
Rat Glucose Surgery
• Intraperitoneal placement
• Occlusion sutures
• Insert sensor
• Seal with Vetbond
Rat Glucose Surgery

52. © 2018 Data Sciences International
Rat Glucose Surgery
• Intraperitoneal placement
• Occlusion sutures
• Insert sensor
• Seal with Vetbond
• Secure with fiber patches
Rat Glucose Surgery

53. © 2018 Data Sciences International
Rat Glucose Surgery
• Intraperitoneal placement
• Occlusion sutures
• Seal with Vetbond
• Secure with fiber patches
• Tack connector
Rat Glucose Surgery

54. © 2018 Data Sciences International
Rat Glucose Surgery
• Intraperitoneal placement
• Occlusion sutures
• Seal with Vetbond
• Secure with fiber patches
• Tack connector
Rat Glucose Surgery

55. © 2018 Data Sciences International
Rat Glucose Surgery
• Intraperitoneal placement
• Occlusion sutures
• Seal with Vetbond
• Secure with fiber patches
• Tack connector
• Secure reference electrode
Rat Glucose Surgery

56. © 2018 Data Sciences International
Large Animal Glucose Surgery

57. © 2018 Data Sciences International
• Implant placement
Large Animal Glucose Surgery

58. © 2018 Data Sciences International
• Implant placement
• Vessel selection
Large Animal Glucose Surgery
Click here to learn more about
large animal glucose telemetry

59. © 2018 Data Sciences International
• Implant placement
• Vessel selection
• Secure sensor
– Miller’s and finger trap
Large Animal Glucose Surgery

60. © 2018 Data Sciences International
Calibration Best Practices
View calibration tech-note

61. © 2018 Data Sciences International
Calibration Best Practices – Timing!

62. © 2018 Data Sciences International
• Mouse → Saphenous vein
https://norecopa.no/films-and-slide-shows/mouse
Calibration Best Practices

63. © 2018 Data Sciences International
• Rat → Tail tip puncture
Calibration Best Practices

64. © 2018 Data Sciences International
• Mouse
– Saphenous vein
• Rat
– Tail tip puncture
• Large animal
– May require anesthesia
or vascular access ports
http://www.norfolkaccess.com
Calibration Best Practices

65. © 2018 Data Sciences International
Combining Additional Endpoints
• Rat
– Glucose and cardiovascular
– HD-S11 & XG

66. © 2018 Data Sciences International
• Large animal
– Glucose and cardiovascular
– M1G
Combining Additional Endpoints
• Rat
– Glucose and cardiovascular
– HD-S11 & XG

67. Dr. Stephanie Simonds
NHMRC/NHF Research fellow
Monash University,
Melbourne, Australia
Megan Fine, DVM
MS, DACVS-LA
Veterinary Surgeon,
Attending Veterinarian
Data Sciences International
#LifeScienceWebinar #ISCxDSI
Thank You
For additional information on the products and applications
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