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M678S4-04-C-S074 Page 3 of24 Section C - DesCriptions and Specifications

STATEMENT OF WORK Cl Statement ofWorlc

CLIN 0001 and Option CLIN 0002 shall be in accordance with the Statement of Work attached to this contract.

\,

M67854-04-C-5074
Page 24 of24

Section J - List of Documents. Exhibits and Other Attachments Exhibit/Attachment Table of Contents

DOCUMENT TYPE
Attachment 1

DESCRIPTION Statement of Work

PAGES 10

DATE

M67854-04-C5074

Attachment 1 Statement of Work

I. Technical

A) Objectivestrasks/Concept. Recent advances in directed energy weapons technology suggests that scalable, non-lethal to lethal force systems may be possible. Such a system would be useful in many environments. Two systems currently under development, active denial and pulsed ener ADS and PEP offer mainly complementary ca acities that could address multi Ie tasks ese tasks include the (b) (2) The full capability of these directed energy systems (DE) are still being explored. At their current stage of development, each system has clear nonlethal (ADS) and lethal (PEP) capacities suitable to the above tasks. Our experiments will examine the feasibility of PEP as a new generation non-lethal wea on. Pulsed ener can be confi ed to roduce lasmas of exce tionall hi ener (b) (2) In the studies described below we will determine the feasibility of using the plasma derived EMP to induce pain suitable to disarm and deter individuals or form barriers to the movement oflarge hostile groups. If successfully deployed, PEP could complement ADS in situations in which the latter is ineffective, less effective, or rone to countermeasures. Many of the countermeasures that might be envisioned against (b) (2) ADS offer opportunities for PEP targeting (via plasma induction or ablation of the defense). Despite these potential advantages, certain special capabilities and features of ADS offer advantages over PEP in many scenarios. Therefore, the systems are complementary.
The efficiency and lethality of PEP weapons systems lll'0 straightforward. The non-ballistic, instantaneous properties of DE make precise tlll'geting a straightforwlll'd matter of line ofsi l. Terrific amounts of ener can be delivered over great Potentially. distances with pinpoint accumc . However, the lication of PEP

(b) (2)

(b) (2)
proposal.
(b) (2)

(b) (2)

(b) (2)

(b) (2) uantify tho. characteristics oflaser induced plasmas created. micro-, nano- ico- and femtosecond lasers of multi Ie

These studies will examine the characteristics of In the studies described below, we will describe investigations that explore the human effects ofLIP. Studies are proposed to detennine the capacity o(b) (2) o evoke pain. These studies will be f_ pain performed, in vitro, where the factors such as distance and orientation can be tightly controlled, and where (b) (2) system components can be isolated for detailed quantitative ",udy. A portion of the investigotions will apply _ t o (b) (2) sensory eell preparations. These. be generated by conventional means. Subsequent studies will use laser-induced and plasmas to create . the characteristics of which will be well defined (b) (2) optimized to produce atraumatic sensory influences.

(b) (2)

wi.

bat activate nociceptor. and tbe extent to whicb this Objective I: To determine the feature. (b) (2) activation Js effective without trau.ma. Pain is a primary component of all NL W. Pain can distract and deter individuals resulting in voluntary immobilization and/or flight. Nociceptors are the fundamental delection component of the pain system. Nociceptors transduce a variety of stimuli (gated ionic current) and then encode the pain signal (action potentials). While the mechanisms are not fulJy understood. ADS operates mainly on the transduction component by heating biological tissue to activate heat transducing proteins at a Techniques for Stimulation ofNociceptors. NTIC proposal, October, 2003). In contrast, _ could activate nociceptors at the level of (b) (2)

M67654-04-C-5074

Attachment 1 Statement of Work

encoding, thereby bypassing the transduction level. Induction at the encoding level is potentially more advantageous. as it avoids the the risk that occurs from this time dependent event. Moreover, by engaging the encoding event, _ (b) (2) will not rely solely on specialized transduction proteins that are selectively expressed in a

subpopulntion of sensory afferents. Although they differ in i."fenn and

th.t medi.te eneoding are

(b) (2) present in all excitable tissue. In objective 1, we will deternllnc the influence nociceptor activation, focusing specifically on cutaneous nociceptors that innervate superficial skin (epidenois) and underlying tissue (demtis). The strength required to induce activation, the contribution of pulse duration and burst frequency will be defined in tightly controlled experiments, in vitro. These data should prove to be very useful in interpreting the potential bwnan effects of LIP, and its potential as a NLW.

Objective 2: To eJ:amJoe tbe influence o f(b) l a s e r plasmas, OD nociceptor activation aod determine tbe _ (2) extent to which tbls activation 10 effective without trauma. Completion ofobjective I will enable a set ofh theses that (b) (2) 2. With an understanding of the 'safe' for (b) (2) _ , directed choices can be made to study particular laser _ (b) (2) configurations on identical recordin methods ut lascr stimulation) we will examine the nociceptor activating properties o f l a s e (2) (b) r _ (b) (2) configumtion and stimulation regimes.

(b) (2)

(b) (2)
B) Background
Laser Plasma Technology. There is increasing interest in the use oflasers for non-conventional defense applications. This is not only a consequence of the recent heightened sensitivities in such areas as homeland security. defense force prolection, and law enforcement, but it also comes from new technical opportunities becoming available through the increasing pace of developments in laser technology. Developments in solid state laser technology in particular are leading these advances. Diode-pumping, for instance, for the first tiroe enables electrical pump energy to be selectively channeled to spccific laser transitions within solid-state laser media, leading to vast improvements in laser efficiency, compactness and stability. New evolutions in laser architecture, like fiber-lasers, slab-laser amplifiers, active phase control and ultra-short pulse technology are rapidly opening up new parameter space in sciences and technologies having possible relevance to new defense applications.
One of these areas is field of laser plusmas.

M61854-04-G-5014 Attachment 1 Statement of Work

(b) (2)

(b) (2)
(b) (2)

(b) (2)
(b) (2)
In the new realm ofhigh intensity femtosecond laser
of laser interaction science has been accessed leadin

(b) (2)

(b) (2)
Assuming that the cro,,-section ofthese highly transient targeL Given that they times measured in picoseconds, the frequency of (b) (2) souree would be in the - " , g i o n . However, to date, little research has verified the

(b) (2)

There is extensive interest in developing weapons systems that utilize pulse energy projectiles (PEP). When appropriately configured, a PEP could serve both lethal and non-lethal lications. The guiding hypothesis of this proposal is that the (b) (2) creation ofLIP can serve as a NLW by activation ofnocicepoors. The Peripheral Pain System. The detection ofpain begins with a complex set ofperipheral afferents (nociceptors) that detect and encode a great variety of stimuli. These peripherally encoded events are relayed by axons into the central nervous system (spinal cord, thalamus, cortex) where the information undergoes the complex assembly required to produce a localized, conscious perception ofpain (Cooper and Sessle, 1993). Nociceptive atferents detect tissue damaging or near tissue damaging consequences ofmechanica! and thermal events, and the chemical events associated with actual tissue damage. To accomplish these multilevel tasks, the pain system has evolved a family ofnociceptive neurons with dive"" mechanical, thermal and chemical response capacities. These capacities overlap in a manner that is not completely understood, but it is likely that they vary for particular tissue sites (skin, joints, muscle, viscera, bone) that have highly specialized nociceptive requirements. Recent advances in nociceptor characterization have permitted classification, In Vi/TO. ofatlcast 8 distinct nociceptive phenotypes. Our laboratory has shown that sensory cells ofthe DRG are comprised ofdiscrete, internally homogenous, classes ofcapsaicin (OC) sensitive (types I, 2, S, 7, 8 and 9) and insensitive (types 3, 4, 6) populations with distinct capacities to respond to SHT, PGEz, protons, ACh and ATP (Martenson et al., 1994; Cardenas et al., 1997; Cardenas et ai, 1999; Petruska et aI., 2000, 2002; Cooper and Cooper, 2001). We have used lipid soluble fluorescent tracers to define the specific distribution of nociceptors into viscera, joints and skin. Prc:liminary studies have indicated ofskin (b) (2) S. It is these nociceptors that are likely to receive the maximal burst _ from laser plasmas _(b) (2) (b) (2)
(b) (2)

M67854-04-C-5074 Attachment 1 Statement of Wor!< The capacity of a nociceptor to detect and transduce noxious stimuli (heat, mechanical, chemical) is due to the presence of membrane imbedded proteins which act as transducers. Specific proteins have evolved which alter conformation in the presence of heat, chemical ageots, (b) (2) This altered conformation gates a pore to allow ions to pass along their

electrochemical gradients. Microwave radiation. via its capacity to beat tissue, is likely to interact with certain heat sensing
proteins that are differentially expressed in nociceptor subpopulations (TRPVI, TRPV2; Caterina et al., 1997, 1999; Tominaga et aI., 1998). Such proteins are likely to be the ultimate targets of ADS millimeter wave radiation. In addition to detection and transduction of noxious events, nociceptors, like alt sensory afferents must encode the event so that it can be relayed to the central nervous system where perceptions are fonned. Each nociceptor emits a code in the form of a series of action potentials that are produced in a frequency that is in proportion to the ionic current of the transduced event. The action potential code arises from the influence of the ionic current on clusters of voltage-gated channels. This can be thought ofas an analog to digital conversion, where the ionic current is the analog signal that is converted to a digital code by the cluster of voltage gated channels. This cluster is composed mainly of volrage gated Na+, K+ and Ca- channels. Each channel is composed of multiple proteins that fann an ionic pore in the neuronal membrane and contain a distinct voltage-sensing region. Sensitivity to internal voltage vanes considerctbly in sensory systems due to the differcntiw distributi<)fi and multiple isoforms of voltage gated channels. Voltage gated channels (No,) are responsible for the upstroke of the action potential while voltage gate K+ channels (1(,) are responsible for the downstroke. Multiple forms of No, and K, have evolved to set characteristic frequency response rates in different afferent populations. Nocice tora contain multi Ie forms of these channels a. 1.7, 1.8 and 1.9; 2002; Djoubri et al., 2003a,b). (b) (2) _ (b) (2) Those Na, subtypes that are mainly foun 1D noclceptors (Na,I.8 and Na,I.9) have relatively high thresholds and slow kinetics. Due to the ultra slow kinetics ofNa,1 .9, only Na.,1.8 participates directly in action potential generation (Elliott and Elliot, 1993; Akopian et al., 1996; Tate et aI., 1998; Cummins et al., 1999; Dib-Hajj et al., 2002).

Na

(b) (2)

(b) (2)

(b) (2)
C) Technical Approach and Methodology

Overview of Experiments. The goal of the studies, in year 1, will be: I) to determine the nano- and micro-pulsed (b) (2) regimes that initiate nociceptor activation; 2) to determine the range of frequency modulation of the nociceptive signal that can be produced; 3) to determine the differential influence on distinct skin nociceptor phenotypes; and 4) to determine the pOint at which trauma might begin to limit the NLW value created. The body of knowled e acquired in year I will guide the development of hypotheses regarding the desired features of a lasma (b) (2) . The e eriments of ear two will test these h otheses using a variety oflasers (b) (2) opefully we will be able to marry these two bodies of knowledge and perfect a laboratory scale NL W laser plasmas. These studies will be conducted in vitro, where nOCiCntiVe cells of several phenotypes can be exposed to well specified, (b) (2) (b) (2) Due to methods developed in our labomtory, we are able to that simulate exposure to laser nociceptive phenotypes that are subpopulations of a large population of sensory cells that mediate touch, proprioception, warmth, cooling, itch and pain sensations (petruska et al., 2000, 2002). The identified nociceptive subpopulations have been shown to be heat sensitive and thereby involved in the transduction ofbuming pain sensations 4

M67854-04C-5074 Attachment 1 Slatement of Work

(b) (2)
In our studies we will present high intensity nanosecond-micosecond pulses to cutaneous nociceptors (b) @...arbocyanine dye tracing). These nociceptors express distinct Na, that are likely to manifest differential sensitivity _ (2) (b) (2) (b) (2)the effuct of repeated _ activation. We will detennine the threshold for activation for nano- and micro- pulsed _ pulsing, pulse duration and intensity. If activation is discrete, we should be able to drive nociceptors in a pulse.by-pulse manner. Alternately, single pulses in these time and intensity domains may not be able to produce any activation. In this (b) (2) instance, burst application that approaches known thresholds o_effectiveness (I msec) could be used. We will conduct such single, multiple and burst train studies at various power and duration combinations in multiple nociceptive subtypes. We will parallel these studies with examinations membrane damage suggestive of electroporatioD, cell trauma and death.

Due to limits of the current technology for delivering high (b) (2) we will not be able to test in the femto- and in year 1. On the one hand, that will limit our ability to form hypotheses that simplify studies of year 2 (b) (2) involving _ single pulse femto- and picosecond lasers. On the other hand, the shoIter the duration of the burst, the less time domains can be likely it will work in single pulse mode. In year 2, (b) (2) can be functionally eXlended examined. We might find that they work in burst mode, where the duration _ into the nanosecond time domain. To that ex.tent, the nanosecond (b) (2)could successfully emulate a burst offemto- and picosecond laser (b) (2) . Femto second lasers have logistic advantages over other configurations.
In year 2, we will use our acquired knowledge of pulse duration, frequency and burst regimes to select laser _(b) (2) with high promise for NLW effectiveness. Based upon studies using a high repetition-rotc (100 Hz) Q-switched Nd: VAG laser and two ad . . that use an 0 n-arcbitecl\m solid-slate oseillator-multi-amplifier system ofour own design, (b) (2) (b) (2) (b) we will bave determined the characteristics _ (2) that best match those propeIties we predict (from year 1) will have atraumatic NLW effec1iveness. In year 2 we will confirm these hypotheses (adjust as necess and examine whether the influence on nod tors are robust with res ct to variations. These variations (b) (2) (b) (2) d include . While the methods of We will again examine neurons for evidence ofdamage due to stimulation differ considerably, the methods of recording from cells will remain the same. Because of the use oflasers in year two, the studies will shift to the University ofCentral Florida site (M. Richardson laboratory). Neural recording equipment will be shipped to the site, and some additional purchases will be made for auxiliary instruments that would be needed at the non-UF location. Nociceptor RecordIngs. Conventional whole cell patch recordings (b) (2) would be desirable and could be made in many of the planned experiments. Thes.alwa. be suitable for classification of nociceptive cells at the beginning ofeach experiment prior to the application of (b) (2) . These methods may be suitable for recording during single or (b) (2) dispersed pulses. However, as the duty cycle or burst frequency increases, the ability to make such recordings by we conventional methods is less likc:ly. Therefore (2) will plWl to use 0 tical methods to assess action potential discharge. (b) Accordingly, cells will be perfused witb dye . Hi resolution (b) (2) (b) (2) is possible under thesc conditions, (b) (2) - - . All recordings are conducted at 35 C.

li

Procedures: Nociceptor Activation. Once the whole cell patch configuration is achieved, cells are classified by physiological criteria associated with nociceptors (voltage clamp mode; see Petruska et a1. 2000, 2002). The main studies arc carried out in current clamp mode. The cell (20-45 urn diameter) is centered in the field (eyepiece reticule). The microscope is configured (b) (2) for -'pplication by the introduction of a pair ofstainless steel plate electrodes tbat have been pre-positioned to bracket the (3 mm, se aration. Durin rccordin SJ cells are exposed to nano- or microsecond pulses from one (b) (2) of the _ (b) (2) pulsers . These devices can produce pulse durations from 10 nanoseconds to 100 microseconds . High exposures are commenced at planned intensities, durations, repetition rates, and burst frequencies. Optical recordings are made continuously and captured by software for analysis, Studies will define the minimum field characteristics that produce activation, and then proceed with higher burst frequencies, longer duralions and more intense fields to determine the limits of activation and thiint at which trauma occurs. (b) (2) conventional records. we will monitor RMP at regular 'rest' intervals. (b) (2) Studies will proceed on a variety of skin nociceptive phenotypes (types 1,2,4 and 5). Differences in susceptibility are likely to be observed due to quantitative and differential expression ofTTX sensitive channels (Na,1.7 vs Na"I.8). We will or ITX (I uM) to determine whethertbe dye emissions are due to gating ofNa,. or a direct influence of (b) (2) dye emission. Some time limited artifact is expected.. If prolonged, false signals are

Un

M67854-04-C-5074

Attachment 1 Statement of Work

indicated, we will shift to Ca++ sensitive dyes. We will also consider thennal contributions by examining the inter-plate temperature shifts associated. with stimulation protocols.

(b) (2)

(b) (2) fields generated. Measurement of nano and micro pulsed E fields. We will (b)- . and also devices developed (2) These will be developments from devices we have used in the pastspecifically for these studies.

The objective of these measurements will be to:

(b) (2)
(b) (2)
(b) (2)

ofthe

(b) (2)

(b) (2)

(b) (2)
(b) (2)

(b) (2)

detectors will be used for measurements _(b) (2) from lasmas created by nanosecond (b) (2) We will also use We will also use more sophisticated signs that have rotection or defense
(b) (2)

here will be to adapt these concepts, and utilize the broad depth ofknowledge in to use with microscopic laserplasma-based sources.

References
Akopian AN, Sivilotti L, and Wood IN. A tetrodotoxin-resistant voltage-gated sodium channel expressed by sensory neurons. Nature 379(6562): 257-262,1996. Cardenas CG, Del Mar LP, Cooper BY, and Scroggs RS. SlIT. receptors couple positively to tetrodotoxin-insensitive sodium channels in a sub 0 ulatioDs ore saicin-sensicive rat senso neurons. Journal 0 Neuroscience 17: 7181-7189, 1997.

(b) (2)
Caterina MI, Rosen TA, Tominaga M, Brake AJ, and Julius D. A capsaicin-receptor homologue with a high threshold for noxious heat. Nature 398: 436-441, 1999.
Caterina MJ, Schumacher HR, Tominaga M, Rosen TA, Levine JD, and Julius D. The capsaicin receptor: a heat

activated ion channel in the pain pathway. Nature 389: 816-824,1997. Cooper, A. and Cooper B.Y. The Distribution ofoAChr in Subclassified Sensory Cells ofthe Rat DRG. Neuroscience Abstracts, 2001, 27, 2001. Cooper, B.Y., Rau, K. and Jobnson, RD.. Heat Reactivity and TRP expression in Capsaicin Sensitive and Insensitive Subclassified Sensory Cells ofthe Rat DRG, Society for Neuroscience 2003. Cooper BY and Sessle BJ. Physiology ofnociception in the trigeminal system. In: The Headaches (I ed.), edited by Olesen J, Tfelt-Hanscn P and Welch KMA. New York: Raven Press Ltd, 1993, p. 87-92. Cummins TR, Dib-Hajj SD, Black lA, Akopian AN, Wood IN, and Waxman SG. A novel persistent tetrodotoxinresistant sodium current in SNSnuli and wild-type small primary sensory neurons. J Neurosci J9: RC43, 1999. Dib-Hajj S. Black JA, Cummins TR, and Waxman SQ. NaNlNavl.9: a sodium channel with unique properties. Trends

Neurosci 25: 253-259, 2002.

M67854-04-C-5074 Attachment 1
Statement of Work

Djouhri L, Fang X, Olruse K, Wood IN, Berry eM, and Lawson SN. The TTX-resistant sodium cbannel Navl.g (SNSIPN3); expression and correlation with membrane properties in rat nociceptive primary afferent neurons. J Physio/550; 739-752,2003a.

Djouhri L, Newton R, Levinson SR. Berry CM, Carruthers B. and Lawson SN. Sensory and electropbysiologicnl

properties of guinea-pig sensory neurones expressing Nav 1.7 (PN1) Na+ channel alpha subunit protein. J Physio/ 546: 565576,2003b.

(b) (2)

Elliott AA and Elliott JR. Characterization ofTIX-sensitive and TTX-resislant sodium currents in small cells from adult rat dorsal root ganglia. J Physio/463; 39-56, 1993. Fang X, Djoubri L, Black lA, Dib.Hajj SD, Waxman SO, and Lawson SN. The presence and role of the tetrodotoxinresistant sodium channel Na v 1.9 in nocice live rim afferent neurons. J Neurosci 22: 7425-7433, 2002.

(b) (2)

Martenson ME, Ingram SL. and Baumann TK. Potentiation of rabbit trigeminal responses to capsaicin in a low pH environment. Brain Research 651: 143-147, 1994.

(b) (2)
(b) (2)

(b) (2)

Tate S. Benn S. Hick C, Trezise D, John V, Mannion RJ, Costigan M, Plumpton C, Grose D, Gladwell Z, Kendall G, Dale K, Bountra S, and Woolf Cl. Two sodium channels contribute to the TTXR sodium current in primary sensory neurons. Nature (Neuroscience t: 653-655,1998.
(b) (2)

(b) (2)
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