Transcript Combined Honey1 Rankin and Hyder.ppt

Making Honey and Medicine
a Local Affair
Dr Stephen Rankin and Dr. Don Hyder
Part 1: The Clinical Trial: Dr. Stephen Rankin (Pinon Family Practice)
Part 2: The Honey Analysis: Dr. Don Hyder (San Juan College)
The Clinical Trial
Approvals
• FDA has approved the project, IND 115653
• SJRMC IRB has approved the Honey Project
• SJC IRB has approved the Honey Project in
relation to the Distinguished Teaching Chair
Award
Clinical Trial Planning Committee
R. Stephen Rankin, M.D., Principal Investigator
Joseph Pope, M.D.
Mary Doshi, Associate Professor, SJC, Medical
Laboratory
Don Hyder, Professor, SJC, Biology
Lynn Lane, Technology Trainer, SJC, Computer Science
Beth Philips, Research Consultant, SJRMC IRB
Administrator
History
• Honey has been used for wound healing for over a thousand
years
• Honey used in wound care comes from Australia and New
Zealand
• Samples from Ireland and Africa tested against CA-MRSA
(community acquired MRSA) isolates and other Staphylococcal
specimens
• Results were favorable in demonstrating activity against CAMRSA
• Little in vitro or in vivo testing of honey from the United States
• Preliminary in vitro studies from Northwest New Mexico honey
demonstrated bactericidal activity against CA-MRSA
• Preliminary screening of the local honey for C. botulinum
Benefits of honey in wound healing
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Moist, antibacterial wound healing environment
Prevents spreading of infection
Renders wound sterile
Rapidly eliminates wound odor
Cleans and debrides wound
Anti-inflammatory
No adverse effects
Diminishes pain
Reduces swelling
Works when other traditional treatments have failed
caMRSA*
• Community acquired Methicillin Resistant Staphylococcus aureus
• Common cause of skin and soft tissue infection
• May become invasive to brain, spinal cord, bone, and other body
tissues
• High incidence of recurrences personal and family
In Vitro Studies
Acquired MRSA
Vancomycin
Manuka Honey
Control
Clover
Local Project
Honey Lot 1
Wildflower
Local Project
Honey Lot 2
Photo courtesy of Mary Doshi ,MLS, (ASCP)© Director,
Program in Medical Technology , San Juan College
Our study
• Further investigate in vitro activity of this New Mexico
varietal honey against CA-MRSA
• Collaboration of Pinon Family Practice, San Juan College,
and SJRMC
• Conduct a controlled clinical research protocol
demonstrating its effectiveness against CA-MRSA
abscesses
• Involve students at San Juan College from multiple
disciplines
Patient population characteristics
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Inclusion:
Ages 16-79
Abscess less than or equal to 6 cm in diameter on arms, legs, abdomen, back
Patient willing to consent and try honey prepared dressing
Patient willing to return to office each day for 7 days
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Exclusion:
Under age 16 or 80 or above
Have underlying immune system disorder
Abscess of face, scalp, breast or genitals, hands or feet
Diagnosed as diabetic
Pregnancy
Allergic to bee pollen, honey, sulfa antibiotics, or lidocaine
Diagnosed with peripheral vascular disease
Recurrent use of alcohol or drugs
Study procedure
• After consent patient receives incision and drainage of abscess and
randomized to :
1)Topical honey
2)Oral antibiotics
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Culture of wound sent to SJRMC and SJC on Day 1
Daily photos of wound
Measurement of wound size
Temperature
Daily cultures of wound sent to SJC
Return daily for 7 days
Goals
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Compare efficacy
Compare side effects
Compare sterilization of wound
Compare patient acceptance of treatments
PRELIMINARY RESULTS
• Wound (Erythema)
• Antibiotic patients (N = 7)
• Average initial size in cm squared 27.9 (range 6.6 to 72.0)
• Average final size in cm squared 9.7 (range 0.0 to 42.0)
• 65% reduction
• Honey patients (N = 4)
• Average initial size in cm squared 16.0 (range 3.2 to 36.0)
• Average final size in cm squared 2.6 (range 0.0 to 9.0)
• 83% reduction
Skin and soft tissue infections
• Current treatment involves incision and drainage with or without
antibiotics
• Role of antibiotics is uncertain with attendant side effects and
resistant organisms
• Recurrent infections with MRSA are significant
• Study by Fritz (Clin. Infect. Dis. 2012:54:743-751) showed a 72%
recurrence rate at 12 months even with decolonizing regimen
• Clearly something else is needed
• Can a topical bactericidal honey have a role in both treatment and
preventing recurrences
The bees
• Treatment Free (No antibiotics or miticides)
• Survivor Queens locally raised and from breeders who use no
treatments
• Forage is varied , along San Juan River near Farmington, New Mexico
• Honey is unheated and unfiltered
Characterization of Antibacterial
Components, Pollen and Nectar Sources
of a Honey Being Used to Treat
Community Acquired MRSA Infections
Bryden Baker, Holly Vandever, Dr. Eric Miller and Dr. Don Hyder
Funding provided by the National Institute of General Medical
Sciences (8 P20 GM 103451) from the National Institutes of
Health
Hypothesis
• Locally produced clinical honey currently being investigated in the
treatment of Community Acquired Methicillin Resistant
Staphylococcus aureus (CA-MRSA) is derived from a single pollen
source (Russian Knapweed) and the primary antibacterial compound
is methylglyoxal.
• Methylglyoxal is known from a medically recognized honey, Medihoney or Manuka Honey, derived from the Manuka plant in New
Zealand.
Objectives
• 1) Identify the primary floral source, (pollen and nectar) of the local
clinical honey
• 2) Identify and quantify a known antibacterial component of honey,
methylglyoxal
• 3) Investigate the antibiotic components of nectar from Russian
Knapweed
Methods (Floral Sources)
• 1) Survey and collect pollen from floral sources in area surrounding
Study Area bee hives
• 2) Prepare and observe pollen using light and electron microscopy
• 3) Extract pollen from clinical honey and observe using light
microscopy
• 4) Quantify pollen in clinical honey using a pollen trap on bee hives
Methods (Chemical Analysis of Clinical Honey, Manuka Honey and
Russian Knapweed Floral Extract)
• 1) Prepare Clinical honey, Manuka honey and Russian Knapweed
floral extract for extraction using Solid Phase Micro-extraction
(SPME) techniques.
• 2) Analysis of extracts using Gas Chromatography-Mass
Spectrometry
Pollen Study Results
Russian Knapweed
Coyote Willow
Musk
Thistle
Fourwing
Saltbush
Coyote
Willow
Russian
Tamarisk
Knapweed
Pollen Grains Using Light Microscopy
Pollen Study
Common
Name*1
Russian
Knapweed
Four-wing
saltbush
Tamarisk
Tansy Mustard
Princess Plume
Russian Olive
Scientific name Pollen Grain
Size (nm)
Pollen Trap
(color and
percent of
pollen balls
per 20 ml
sample)
Pollen
Identified in
Clinical
Honey
Centaurea repens
40
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Atriplex canescens
30
Brown/Tan37%
Bright Yellow
14%
Rust- 2%
17-20
40
20-25
White 32%
*
40
Dark Yellow
15% and
Orange-0.5%
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Tamarix parviflora
Descurinia pinnata
Stanleya pinnata
Elaeagnus
angustifolia
Coyote Willow
Salix exigua
Showy Milkweed Asclepias speciosa
Curly Cup
Grindelia
Gumweed
squarrosa
Musk Thistle
Carduus nutans
*1:
20
40
20-40
40
Pollen collected from plants blooming in the vicinity of the bee
hives at the same time as the Russian Knapweed.
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Pollen Study Summary
The clinical honey was identified as a multiple floral source
honey with Russian Knapweed pollen and Coyote Willow
pollens being significant components along with four other
pollen sources.
Chemical Analysis Results
Manuka Honey
Compounds
Methylglyoxal
Acetic anhydride
1,2-Propanediol
1,3-Dioxolane-2-methanol
4-Methyl-1,6-heptadien-4-ol
Methyl 6-oxoheptanoate
2,2-Dimethyl-1,3-dioxan-4-yl ethanol
Silane
4-Hexenal
Hexanedioic acid
Diethyl 1,2-cyclopropanedicarboxylate
D-allo-dec-2-enonic acid
1-Methyl-1-6-ethyl-3-octyloxy-1-silacyclohexane
RT
2.32
3.19
3.91
4.98
5.55
5.64
6.71
7.24
8.16
8.90
9.34
9.56
10.63
No MW (g/mol)
1
72.06
2
102.09
3
76.09
4
104.10
5
126.19
6
158.19
7
160.21
8
32.12
9
98.00
10
146.14
11
186.21
12
372.10
13
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Clinical Honey
Compounds
RT No MW (g/mol)
Methylglyoxal
2.07 1
72.06
Furfural
3.83 2
96.09
1,3,5-Cycloheptatriene
5.62 3
92.14
4H-Pyran-4-one
6.49 4
96.08
2-Furancarboxaldehyde 7.16 5
96.09
1-Dodecanol
7.72 6
186.34
Morphinan-3,14-diol
7.81 7
327.46
Glucopyranoside
8.88 8
194.18
Russian Knapweed Floral Extract
Compounds
RT
No
MW (g/mol)
R-(-)-2-Amino-1-propanol
1.34
1
Topotecan
2.08
2
457.9
Benzenemethanol, a-(15.57
5
aminoethyl)-,[S-(R*,R*)]2,5-Furandicarboxaldehyde
5.89
7
N-Benzoyl-dl-alanine
6.60
8
1,1’-(4-Methyl-1,37.12
9
phenylene)bis[3-(5-benzyl1,3,4-thiadiazol-2-yl)urea
1-(+)-Ascorbic acid 2,68.20
12
dihexadecanoate
Pregan-20-one, 2-hydroxy8.63
13
5,6-epoxy-15-methyl9,12,15Octadecatrienoicacid 8.80
14
Retinal 9-cis-
9.12
15
Chemical Analysis Results Summary
• 1) Methylglyoxal was identified in the clinical honey and in the
Manuka, Medi-honey.
• 2) A secondary compound, furfural, a phenolic compound having
known antibacterial and antifungal properties was identified in the
local clinical honey.
• 3) A known anticancer compound, topotecan (A topoisomerase
inhibitor), was identified in the Russian Knapweed floral extract.
Antibacterial compounds were not found in the Russian Knapweed
floral extract.
Further Research
• Continued Investigation of Floral source(s) of this local New Mexico
honey
• Chemical analysis and in-vitro studies of other local and regional
honey samples against MRSA
• Further clinical research involving bactericidal honey and MRSA
recurrences
• Quantify chemical components of the honey such as methylglyoxal,
furfural and bee definsin-1 levels
Contact Information
• Dr. Stephen Rankin: [email protected]
• Dr. Don Hyder: [email protected]