Funis Vermis, Rope worm - MMS-Seminar.com · Hiromi Shinya. Professor Alex A. Volinsky, Ph.D....

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky Funis Vermis, Rope worm

Transcript of Funis Vermis, Rope worm - MMS-Seminar.com · Hiromi Shinya. Professor Alex A. Volinsky, Ph.D....

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Funis Vermis, Rope worm

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Funis Vermis, Rope worm

2 cm

Worm is slimy, twisted like a rope, has a specific smell

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DAPI Stain

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Successful DNA Extraction

DNA #Tissue

Amount (g)

Total

Volume

(ul)

Total Quantity

(ug)

Concentration

(ug/ul)

Volume

per 0.1ugRatio

Qubit Total

Quantity

(ng)

Qubit

Concentration

(ng/ul)

1 Stage -3 0.2 100 7.74 0.077 1.29 1.548 424.000 4.24

2 Stage -5 0.2 100 7.89 0.079 1.27 1.579 552.000 5.52

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Sequencing details

First, DNA was extracted from the worm tissue using Qiagen DNeasy mini kit, and

the quality was verified by agarose gel electrophoresis. From ten nanograms of

genomic DNA, the primer pair LCO1490 (5'-GGTCAACAAATCATAAAGATATTGG-

3') AND HCO2198 (5'-TAAACTTCAGGGTGACCAAAAAATCA-3') was used to

PCR-amplify a 658 bp region (Folmer, et al) of the mitochondrial cytochrome c

oxidase subunit I gene using the Epicentre Failsafe PCR Kit using PCR Buffer E

(Epicentre Biotechnologies, Madison, WI). Each 25 ul PCR reaction contained 1

uM of each primer, 0.625 Units FailSafe PCR Enzyme Mix, and 20 ng of template

DNA. PCR cycling conditions were as follows: 3 min at 95 °C, followed by 35

cycles of 95 °C for 50 sec, 55 °C for 1.5 min, and 72 °C for 1.5 min, and a final

extension of 72 °C for 8 min. The PCR product was cleaned up with ExoSAP-It (Affymetrix, Inc., Santa Clara, CA) and the same primers two sequence in each

direction using the ABI3130xl Genetic Analyzer 16-capillary sequencer with the

BigDye v3.1 chemistry (Life Technologies, Grand Island, NY).

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Cytochrome oxidase subunit I, COIStage 3 - Forward Read. Human presudogene, chromosomes 8, 17. Cymoninus notabilisAGTCTTGGGCACAGCCTTAAGCCTCCTTATTCAAACTGAACTAGGTCAACCTGGCAACCTTCTAGGTAACGACCACATCTACAACGTCATCGTCACAGCCCA

CACATTCTTCATAATCTTTTTCATAGTAATGTCTATCATAATTGGAGGTTTTGGCAATTGACTGGTCCCCCTGATAATTGGCGCCCCTGACATGGCATTTCCC

TGCGTAAATAACATAAGCTTCTGGCTCCTTCCCCACCCCTTTCTACTACTGCTGGCATCTGCTATGATAGAAGCTGGTGCCGGAACAGGTTGAACGGTTTAC

CCTCCTTTGGCAGGAAACTATTCCCATCCAGAAGCCTCTGTAGACCTAACCATTTTCTCCCTTCATCTAGCAGGTGTCTCCTCTATTCTAGGAGCTATTAATT

TCATCACCACAATTATTAACATAAAACCCCCTGCCATATATCAATACCAAACGCCCCTCTTTGTCTGATCTGTCCTAATCACGGCAGTCCTACTCCTCCTCTC

CCTTCCAGTCTTGGCCGCTGGCATTACTATACTGCTAACAAACCGTAACCTTAATATTACCTTTTTGACCTGGCCGGAGGAGGGGACCCTATCCTATAACAA

CACCTATTCTGATTTTTGGTC

Stage 3 - Reverse ReadCTCCTCCGGCCAGGTCAAAAAGGTGATATGTTGGTTACGGTCTGTTAGCAGTATATTAATGCCAGCGGCCAAGACTGGAAGGGAGAGGAGGAGTAGGACT

GCCGTGATTAGGACAGATCAGACAAAGAGGGGCGTTTGGTATTGATATATGGCAGGGGGTTTTATGTTAATAATTGTGGTGATGAAATTAATAGCTCCTAGA

ATAGAGGAGACACCTGCTAGATGAAGGGAGAAAATGGTTAGGTCTACAGAGGCCTCTGGATGGGAATAGTTTCCTGCCAAAGGAGGGTAAACCGTTCAACC

TGTTCCGGCACCAGCTTCTATCATAGCAGATGCCAGCAGTAGTAGAAAGGGGTGGGGAAGGAGCCAGAAGCTTATGTTATTTACGCAGGGAAATGCCATGT

CAGGGGCGCCAATTATCAGGGGGACCAGTCAATTGCCAAAACCTCCAATTATGATAGACATTACTATGAAAAAGATTATGAAGAATGTGTGGGCTGTGACGA

TGACGTTGTAGATGTGGTCGTTACCTAGAAGGTTGCCAGGTTGACCTAGTTCAGTCTGAATAAGGAGGCTTAAGGCTGTGCCCAAGACTCCAGCTCATGCA

CCAAACAATAAATATAGTGTTCCAATATCTTTATGATTTGTT

Stage 5 - Forward Read. Human mtDNAGTCCTAGGCACAGCTCTAAGCCTCCTTATTCGAGCCGAGCTGGGCCAGCCAGGCAACCTTCTAGGTAACGACCACATCTACAACGTTATCGTCACAGCCCA

TGCATTTGTAATAATCTTCTTCATAGTAATACCCATCATAATCGGAGGCTTTGGCAACTGACTAGTTCCCCTAATAATCGGTGCCCCCGATATGGCGTTTCCC

CGCATAAACAACATAAGCTTCTGACTCTTACCTCCCTCTCTCCTACTCCTGCTCGCATCTGCTATAGTGGAGGCCGGAGCAGGAACAGGTTGAACAGTCTA

CCCTCCCTTAGCAGGGAACTACTCCCACCCTGGAGCCTCCGTAGACCTAACCATCTTCTCCTTACACCTAGCAGGTGTCTCCTCTATCTTAGGGGCCATCA

ATTTCATCACAACAATTATCAATATAAAACCCCCTGCCATAACCCAATACCAAACGCCCCTCTTCGTCTGATCCGTCCTAATCACAGCAGTCCTACTTCTCCT

ATCTCTCCCAGTCCTAGCTGCTGGCATCACTATACTACTAACAGACCGCAACCTCAACACCACCTTCTTCGACCCCGCCGGAGGAGGAGATCCCATTCTAT

ACCAACACCTATTCTGATTTTTGGTCAC

Stage 5 - Reverse ReadCTCCTCCTCCGGCGGGGTCGAAGAAGGTGGTGTTGAGGTTGCGGTCTGTTAGTAGTATAGTGATGCCAGCAGCTAGGACTGGGAGAGATAGGAGAAGTAG

GACTGCTGTGATTAGGACGGATCAGACGAAGAGGGGCGTTTGGTATTGGGTTATGGCAGGGGGTTTTATATTGATAATTGTTGTGATGAAATTGATGGCCC

CTAAGATAGAGGAGACACCTGCTAGGTGTAAGGAGAAGATGGTTAGGTCTACGGAGGCTCCAGGGTGGGAGTAGTTCCCTGCTAAGGGAGGGTAGACTGT

TCAACCTGTTCCTGCTCCGGCCTCCACTATAGCAGATGCGAGCAGGAGTAGGAGAGAGGGAGGTAAGAGTCAGAAGCTTATGTTGTTTATGCGGGGAAAC

GCCATATCGGGGGCACCGATTATTAGGGGAACTAGTCAGTTGCCAAAGCCTCCGATTATGATGGGTATTACTATGAAGAAGATTATTACAAATGCATGGGCT

GTGACGATAACGTTGTAGATGTGGTCGTTACCTAGAAGGTTGCCTGGCTGGCCCAGCTCGGCTCGAATAAGGAGGCTTAGAGCTGTGCCTAGGACTCCAG

CTCATGCGCCGAATAATAGGTATAGTGTTCCTATATCTTTATGAGTTTGTTAACCA

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Cymoninus notabilis, a seed bug. 98% COI match

Caribbean insects @ Harvard Entomology

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Phylogenetic tree. Stage 3

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Optical microscopy

40 µµµµm

C D

12 µµµµm

B

12 µµµµm

A

90 µµµµm

Gas bubbles present in microchannels

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Environmental SEM

Microchannels terminating on the rope worm surface

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Earlier stages. Gas bubbles

5 cm

AB

3 cm

C

1 cm

Fecal Stone

BubblesWorm head with blood

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Stage 3. Branched jellyfish

2 cm

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Stage 2. Viscous mucus with bubbles

a) b)

3 cm

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5th day of fasting

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Lemon juice enema results

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Fecal Stones with seeds

1 cm

Fecal Stone

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Lymphoid aggregates in Colonoscopy

Colonoscopies on youtube.com show rope worms

Hiromi Shinya

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Mucus in Bronchial Tubes

Bronchial endoscopy on youtube.com shows mucus, which looks similar to rope

worm earlier stages

http://www.youtube.com/watch?v=RUAY8FpgiZQ

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Nasal Endoscopic SurgeryRight Left

Nasal endoscopy on youtube.com shows mucus with gas bubbles, which looks

similar to the rope worm

http://www.youtube.com/watch?v=YPcdI2c9cK0

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Florescent die

http://www.youtube.com/watch?v=P6n5ZREKLRo

Gas bubbles in nasal polyps look similar to gas bubbles in rope worms

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Rope worm in the nose

http://www.youtube.com/watch?v=dDilFvAgdco

Cultured as

Staphylococcus

aureus

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Nasal Polyps

Nasal polyps look similar to the rope

worm. Similar gas bubbles.

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Endoscopic Nasal Polypectomy

Endoscopic Nasal Polypectomy

http://www.youtube.com/watch?NR=1&v=PcK1sBmbFVk&feature=endscreen

Sinusitis, Mucocele, Cyst in the Ethmoid Sinus

http://www.youtube.com/watch?v=PonTWaAXlZg

Endoscopic Sinus Surgery (Dr. Rey Caro)

http://www.youtube.com/watch?feature=endscreen&NR=1&v=OUEYJuhd2uU

Endoscopic Drainage of a Type 2 Hypoplastic Maxillary Sinus

http://www.youtube.com/watch?v=DQDzfSMmMVc

Endoscopic Drainage of an Orbital Mucocele

http://www.youtube.com/watch?v=YNTQKnJ6pxM

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Appendiceal Mucocele

http://www.youtube.com/watch?v=AEuodon3tQM

Appendiceal mucocele looks similar to earlier stages of the

rope worm development. Similar gas bubbles inside.

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Appendix Stones

http://www.youtube.com/watch?v=e992ZVgAbqI

Fecal stones inside of inflamed appendix look similar to

fecal stones produced by the rope worms

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Russian Forums

http://www.pravdu.net/publ/zdorove/parazity_v_tele_cheloveka_otravljajut_mysli_l

judej/2-1-0-111

http://aum.mybb.ru/viewtopic.php?id=633

http://www.web-zdrav.ru/index.cgi/forum?forum=0&topic=13688

Multiple photos of eucalyptus-lemon cleansing results

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

More Russian Forums

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

US Forumshttp://curezone.com http://mmsautism.org

http://www.myjourneywithcandida.com/

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More US pictures

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More

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Fecal Stones

Andreas Kalcker, www.mmsautism.org

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Autistic child

Dr. Andreas Ludwig Kalcker,

http://www.ustream.tv/recorded/28979600

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Ascaris suum and Candida

At www.mmsautism.org rope worms are considered to be ascaris suum remains

and mucus is considered to be candida

http://www.myjourneywithcandida.com/

“Candida, Kind of looks like a jelly like mucus”

http://www.myjourneywithcandida.com/2010/08/pictures-of-candida-what-does-

candida.html

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

And more

Dr. Andreas Ludwig Kalcker, http://www.ustream.tv/recorded/28979600

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

http://www.myjourneywithcandida.com/2010/10/pictures-of-candida-in-my-stomach.html

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Professor Alex A. Volinsky, Ph.D. www.eng.usf.edu/~volinsky

Volgograd Bureau of Forensic Expertise

74 y.o. male, myocardial infarction (heart attack) 70 y.o. male, pneumonia

So far 5 cadavers inspected, and all had fumis virmis in the large intestine