1. A method of treating or preventing toxicity associated with exposure to an organophosphate agent in a mammal comprising administering to the mammal a cationic liposome complex, wherein the cationic liposome complex comprises:
(a) a cationic liposome;
(b) a ligand directly complexed with, but not chemically conjugated to, the cationic liposome;
(c) a nucleic acid molecule encoding butyrylcholinesterase (BChE) associated with the cationic liposome; and
(d) a nucleic acid molecule encoding a polyproline rich peptide associated with the cationic liposome.
2. (canceled)
3. (canceled)
4. The method of claim 1, wherein the complex is administered via a route selected from the group consisting of intranasal administration, intravenous administration, oral administration, sublingual administration, intramuscular administration, intralesional administration, intradermal administration, transdermal administration, intraocular administration, intraperitoneal administration, percutaneous administration, aerosol administration, intraorgan administration, intracereberal administration, topical administration, subcutaneous administration, endoscopic administration, slow release implant, administration via an osmotic or mechanical pump and administration via inhalation.
5. (canceled)
6. The method of claim 1, wherein the ligand is selected from the group consisting of transferrin, an antibody and an antibody fragment.
7. The method of claim 1, wherein the ligand is a single chain Fv antibody fragment.
8. The method of claim 7, wherein the single chain Fv antibody fragment is an anti-transferrin receptor single chain Fv (TfRscFv).
9. The method of claim 1, wherein the ligand-targeted cationic liposome further comprises a peptide comprising a KK(H)KKK5-K(H)KKC (HoKC) (SEQ ID NO: 1) peptide associated with the cationic liposome.
10. The method of claim 1, wherein the nucleic acid molecule encoding BChE is contained in a first plasmid and the nucleic acid molecule encoding the polyproline rich peptide is contained in a second plasmid, or wherein the nucleic acid molecule encoding the BChE and the nucleic acid encoding the polyproline rich peptide are contained in the same plasmid.
11. (canceled)
12. (canceled)
13. The method of claim 1, wherein the BChE is a mutant version of BChE.
14. The method of claim 13, wherein the mutant version of BChE is a G117H mutant.
15. The method of claim 1, wherein the cationic liposome comprises a mixture of one or more cationic lipids and one or more neutral or helper lipids
16. The method of claim 1, wherein the ligand and the cationic liposome are present at a ratio in the range of about 1:1 to about 1:100 (w:w), or about 1:10 to about 1:50 (w:w) or about 1:20 to about 1:40 (w:w).
17. (canceled)
18. (canceled)
19. The method of claim 1, wherein said cationic liposome comprises a mixture of dioleoyltrimethylammonium phosphate with dioleoylphosphatidylethanolamine and cholesterol; a mixture of dioleoyltrimethylammonium phosphate with cholesterol, a mixture of dimethyldioctadecylammonium bromide with dioleoylphosphatidylethanolamine and cholesterol, a mixture of dimethyldioctadecylammonium bromide with dioleoylphosphatidylethanolamine, a mixture of dimethyldioctadecylammonium bromide with cholesterol, or a mixture of dioleoyltrimethylammonium phosphate with dioleoylphosphatidylethanolamine.
20. The method of claim 1, wherein the nucleic acid molecules are present at a molar ratio of about 10:1 to about 1:10 (moles nucleic acid molecule encoding butyrylcholinesterase (BChE): moles nucleic acid molecule encoding a polyproline rich peptide), or about 5:1 to about 1:5 (moles nucleic acid molecule encoding butyrylcholinesterase (BChE): moles nucleic acid molecule encoding a polyproline rich peptide), or about 4:1 (moles nucleic acid molecule encoding butyrylcholinesterase (BChE): moles nucleic acid molecule encoding a polyproline rich peptide), or about 2:1 (moles nucleic acid molecule encoding butyrylcholinesterase (BChE): moles nucleic acid molecule encoding a polyproline rich peptide), or about 1:1 (moles nucleic acid molecule encoding butyrylcholinesterase (BChE): moles nucleic acid molecule encoding a polyproline rich peptide).
21. (canceled)
22. (canceled)
23. (canceled)
24. (canceled)
25. The method of claim 1, wherein the total amount of nucleic acid molecules is present at a weight ratio of between about 1:1 to about 1:40 (\u03bcg nucleic acid:\u03bcg liposome), or at a weight ratio of between about 1:5 to about 1:20 (\u03bcg total nucleic acid:\u03bcg liposome), or at a weight ratio about 1:10 (\u03bcg total nucleic acid:\u03bcg liposome).
26. (canceled)
27. (canceled)
28. (canceled)
29. The method of claim 1, wherein the complex is administered so as to treat toxicity associated with exposure to at least 1\xd7LD50 of the organophosphate agent, or exposure to up to 5\xd7LD50 of the organophosphate agent.
30. (canceled)
31. (canceled)
32. (canceled)
33. The method of claim 29, wherein the complex is administered immediately after exposure to the organophosphate agent.
34. The method of claim 29, wherein the complex is administered at least 6 hours prior to potential exposure to the organophosphate agent.
35. The method of claim 29, wherein the complex is administered at least once a week prior to potential exposure to the organophosphate agent.
36. The method of claim 1, wherein the mammal is a human.
37. A method of treating or preventing toxicity associated with exposure to an organophosphate agent in a human comprising administering intranasally or via aerosol inhalation to the human a cationic liposome complex, wherein the cationic liposome complex comprises:
(a) a cationic liposome;
(b) an anti-transferrin receptor single chain Fv (TfRscFv) directly complexed with, but not chemically conjugated to, the cationic liposome; and
i. a nucleic acid molecule encoding butyrylcholinesterase (BChE) contained in a first plasmid associated with the cationic liposome; and a nucleic acid molecule encoding a polyproline rich peptide contained in a second plasmid associated with the cationic liposome,
or
ii. a nucleic acid molecule encoding butyrylcholinesterase (BChE) and a nucleic acid molecule encoding a polyproline rich peptide contained in the same plasmid associated with the cationic liposome,
wherein the TfRscFv and the cationic liposome are present at a ratio in the range of about 1:20 to about 1:40 (w:w) and the nucleic molecules are present at a ratio of about 1:5 to about 1:20 (\u03bcg nucleic acid:\u03bcg liposome),
and wherein the complex is administered so as to treat or prevent toxicity associated with exposure to at least 1\xd7LD50 of the organophosphate agent.
38. (canceled)
39. (canceled)
40. (canceled)
41. The method of claim 37, wherein the BChE is a mutant version of BChE.
42. The method of claim 41, wherein the mutant version of BChE is a G117H mutant.
43. The method of claim 37, wherein the cationic liposome comprises a mixture of one or more cationic lipids and one or more neutral or helper lipids
44. The method of claim 43, wherein said cationic liposome comprises a mixture of dioleoyltrimethylammonium phosphate with dioleoylphosphatidylethanolamine and cholesterol; a mixture of dioleoyltrimethylammonium phosphate with cholesterol, a mixture of dimethyldioctadecylammonium bromide with dioleoylphosphatidylethanolamine and cholesterol, a mixture of dimethyldioctadecylammonium bromide with dioleoylphosphatidylethanolamine, a mixture of dimethyldioctadecylammonium bromide with cholesterol, or a mixture of dioleoyltrimethylammonium phosphate with dioleoylphosphatidylethanolamine.
45. The method of claim 37, wherein the nucleic acid molecules are present at a molar ratio of about 10:1 to about 1:10 (moles nucleic acid molecule encoding butyrylcholinesterase (BChE): moles nucleic acid molecule encoding a polyproline rich peptide), or at a molar ratio of about 5:1 to about 1:5 (moles nucleic acid molecule encoding butyrylcholinesterase (BChE): moles nucleic acid molecule encoding a polyproline rich peptide), or at a molar ratio of about 4:1 (moles nucleic acid molecule encoding butyrylcholinesterase (BChE): moles nucleic acid molecule encoding a polyproline rich peptide), or at a molar ratio of about 2:1 (moles nucleic acid molecule encoding butyrylcholinesterase (BChE): moles nucleic acid molecule encoding a polyproline rich peptide), or at a molar ratio of about 1:1 (moles nucleic acid molecule encoding butyrylcholinesterase (BChE): moles nucleic acid molecule encoding a polyproline rich peptide).
46. (canceled)
47. (canceled)
48. (canceled)
49. (canceled)
50. (canceled)
51. The method of claim 37, wherein the complex is administered so as to treat toxicity associated with exposure to up to 5\xd7LD50 of the organophosphate agent, or to prevent toxicity associated with exposure to up to 5\xd7LD50 of the organophosphate agent.
52. (canceled)
53. The method of claim 37, wherein the complex is administered immediately after exposure to the organophosphate agent.
54. The method of claim 38, wherein the complex is administered at least 6 hours prior to potential exposure to the organophosphate agent.
55. The method of claim 38, wherein the complex is administered at least once a week prior to potential exposure to the organophosphate agent.
56-69. (canceled)
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A multi-pod conferencing system for local conference participants to communicate with remote conference participants, the multi-pod conferencing system comprising;
a base unit including a base controller configured to communicate audio signals with the remote conference participates over a carrier medium; and
a plurality of pods separate from, and operably coupled to, the base unit, wherein each pod of the plurality of pods comprises:
a pod processor;
a loudspeaker operably coupled to the pod processor;
a digital to analog converter operably coupled to the loudspeaker, the digital to analog converter configured to convert digital audio to analog audio for sound reproduction by the loudspeaker;
a plurality of physical microphones connected to a pair of audio ports with each audio port being equal distance from the loudspeaker;
an analog to digital converter operably coupled with the plurality of physical microphones, the analog to digital converter configured to convert the analog audio to digital audio for processing by the pod processor; and
a plurality of virtual microphones for reducing common-mode noise, each virtual microphone of the plurality of virtual microphones formed from a combination of at least two outputs from the plurality of the physical microphones,
wherein the analog to digital converters for each pod of the plurality of pods are configured for synchronized sampling with the analog to digital converters of the other pods of the plurality of pods in the multi-pod conferencing system.
2. The multi-pod conferencing system of claim 1, wherein the carrier medium that comprises at least one of a telephone line and a voice over internet protocol network.
3. The multi-pod conferencing system of claim 2, wherein the plurality of virtual microphone further provide common-mode noise rejection.
4. The multi-pod conferencing system of claim 1, wherein pod processor is configured to support full duplex operation.
5. The multi-pod conferencing system of claim 1, wherein each pod of the plurality of pods makes a contribution to a summed audio output starting at the last pod in a chain for the plurality of pods and ending in the base unit.
6. The multi-pod conferencing system of claim 1, wherein the pod controller comprises a plurality of distributed processors that are configured to separately control audio and control functions of the pod.
7. The multi-pod conferencing system of claim 1, wherein the plurality of physical microphones are selected from the group consisting of unipolar microphones and bipolar microphones.
8. The multi-pod conferencing system of claim 1, wherein a polarity of the physical microphones are the same.
9. The multi-pod conferencing system of claim 1, wherein the base unit is configured to perform echo cancellation of local audio.
10. The multi-pod conferencing system of claim 1, wherein the pod processor is further configured to perform:
pod echo cancellation for sound between the virtual microphone and said loudspeaker; and
microphone gating for determining the best virtual microphone to gate on or off by using a loudness value, a quietness counter, and a noise floor value, wherein a constant gain is maintained by attenuation being applied to each gated microphone if more than one microphone is gated on at the same time.
11. A method of manufacturing a virtual microphone system for reducing common-mode noise for an individual pod that is part of a multi-pod conferencing system, the method comprising:
providing a base unit having a base processor for communicating audio signals to remote conference participants; and
providing a plurality of pods that are coupled to, and separate from, the base unit, wherein providing the plurality of pods comprises:
providing a pod processor for providing audio processing for various pod functions;
coupling a loudspeaker to a digital to analog converter and the pod processor, the digital to analog converter configured to convert digital audio from the pod processor to analog audio to the loudspeaker;
coupling an analog to digital converter to a plurality of physical microphones that are configured to sample analog audio in a synchronized manner with sampling performed by analog to digital converters of other pods of the virtual microphone system;
connecting the plurality of physical microphone a plurality of audio ports with each audio port being equal distance from the loudspeaker; and
coupling the physical microphones with the pod processor, such that outputs from the plurality physical microphones are combined forming a plurality of virtual microphones for reducing common-mode noise.
12. The method of claim 11, further comprising coupling the base unit with a carrier medium that comprises at least one of a telephone line and a voice over internet protocol network.
13. The method of claim 11, wherein forming the plurality of virtual microphones comprises providing common-mode noise rejection.
14. The method of claim 11, wherein providing a pod processor includes a pod processor configured to perform echo cancellation of sound between the plurality of virtual microphones and the loudspeaker, and perform determining the best virtual microphone to gate on or off by using a loudness value, a quietness counter, and a noise floor value.
15. A method of using a virtual microphone system for a multi-pod conferencing system, the method comprising:
communicating local audio to remote participants through a base unit and coupled with a plurality of pods, wherein communicating further comprises:
audio processing using a pod processor;
converting digital audio to analog audio with a loudspeaker that coupled to a digital to analog converter and the pod processor;
converting sampled analog audio to digital audio using an analog to digital converter coupled to a plurality of physical bi-polar microphones positioned in the horizontal resting plane of each individual pod of the plurality of pods, each individual physical microphone connected to a pair of audio ports with each audio port being equal distance from the loudspeaker, the physical microphones coupled to the pod processor, wherein the sampled analog audio is synchronized with sampling performed by other analog to digital converters from other pods of the virtual microphone system; and
creating a plurality of virtual microphones for reducing common-mode noise, the plurality of virtual microphones are formed by combining outputs from the plurality of physical microphones.
16. The method of claim 15, further comprising a carrier medium that comprises a telephone line, wherein the base unit comprises a base echo cancellation means for echo cancellation of local audio.
17. The method of claim 15, wherein the virtual microphone means further comprises providing common-mode noise rejection.
18. The method of claim 15, wherein audio processing using the pod processor includes:
echo cancelling sound using pod echo cancellation means for echo cancellation of sound between the virtual microphone and the loudspeaker; and
determining the best virtual microphone to gate on or off using a loudness value, a quietness counter, and a noise floor value.
19. The method of claim 15, further comprising communicating audio data between the pod and the base unit.
20. The method of claim 19, wherein communicating between the pod and the base unit employs spread spectrum techniques to the audio data.