The Consultant:  Nematode Infestation

by Duane B. Lakings, Ph.D.

What am I doing wrong?   I mused after another uneventful and unproductive day.  Sitting at my desk with my feet resting on the edge, I waited for, longed for, the phone to ring.  Maybe I should have stayed at the bench and never opened this consulting business.  I pondered.  At least then I was doing science and not just wishing I could do science.  If I don't get a contract or two soon, I'll have to find a job flipping hamburgers or driving a truck or being a private detective.  After taking a sip of lukewarm and too strong coffee from the almost empty cup, I filled my favorite pipe with black Cavendish and lit it.  The white smoke curled up to the ceiling.  While staring at my blank computer screen, I continued to contemplate my bad, and getting worse, situation.

The late afternoon sun filtered through the window and a chilly autumn breeze ripped the few remaining leaves from the trees.  After running fingers through thick, mostly brown hair and pulling on the whiskers of a partially gray, full bread, I slowly lowered my feet and picked up the phone, quickly punching in a number.  I needed to talk with someone and who better than an old friend.  "Hello, this is Dr. David Wilcott.  I'd like to speak to Dr. Betty Mills,"  I said to the administrative assistant who answered the phone.

A few moments later, a soft, familiar voice answered,  "David, I was just thinking about you.  Did you get the e-mail note I sent?"

"I haven't read my mail for a couple of hours."  I wished I had checked before calling Betty.  "Was it important?"

"If you're interested in a little work,"  Betty chuckled,  "I guess it could be considered important."

I sat up straight and took a calming breath.  "Of course I'm interested in some work.  What do you need?"  In my mind, I pictured Betty in her white lab coat, her long, auburn hair shining in the fluorescent light.  She is a molecular biologist, one of the best I have ever known.  She complimented my meager, in my opinion, abilities as a bioanalytical chemist.

"We have a problem here and I think we could use your expertise."

"Always ready to help you, Betty."  I hoped whatever my former collaborator had in mind would provide me with enough capital to keep my struggling business open.  "Just like old times."  We had worked together at Hansen Pharmaceuticals to develop a number of drug candidates and to show how an even greater number did not have the necessary attributes to become products.  After a merger with Stebark International, we had been downsized or rightsized or steamlined or decruited or any other term used to indicate that we were fired.  I opened my consulting business and Betty moved to a small agriculture biotech firm where she was heavily involved in finding ways to increase crop production.  "What type of mess have you gotten yourself into this time?"  I said lightly.

I heard her laugh slightly and knew her light blue eyes were sparkling.  "David,"  she said softly,  "do you recall the many heated discussions we had on the scientific results of discovery leads before they became potential drug candidates?"

"Of course, I do.  They were the highlight of my day, even if we didn't always agree."

"We have a similar situation with one of our programs.  Probably our largest and most important project.  I could use your advice."

Betty's voice displayed some tension and I knew she was not happy with whatever was happening.  "Tell me more and I'll give you my opinion.  First half hour is free.  After that, I'll charge you an arm and leg."  I hoped the discussion would not be a free consultation.  However, if Betty needed my help, I would do my best, even for nothing.  That's what friends are for, isn't it?

"As I mentioned in my note,"  Betty said, carefully choosing her words,  "we've been generating data on a nematode that was accidentally introduced into the Midwest with some ornamental grass plants imported from Asia.  We, and the rest of the world, are finding that many of these unwanted species, or exotics, can become major problems when they're able to adapt to their new environment.  Most of them don't have nature's normal checks and balances in place.  So when they break out, there is nothing to stop them."

"Nematode?"  I muttered, struggling to remember what I had read concerning these little animals in one of the many scientific journals I skimmed while searching for potential consulting leads.  "That's a type of roundworm, isn't it?"

"I'll make a biologist out of you yet, David."  Betty chuckled.  We had often discussed my limited knowledge in biology and her less than complete understanding of the physical sciences, which is one reason we worked together so well.  We approached problems from different directions and our expertises were often synergistic in finding potential solutions.  "This little beast has no known predators in the Midwest.  That in itself was not a major problem as long as the worms were localized to these ornamental plants.  Some farmers decided to spruce up their homes and planted the grass too close to their cornfields.  They quickly learned that these nematodes love corn."

"So do I.  Especially on the cob with lots of butter."  I glanced down at my protruding stomach, confirming my statement.  "Does that mean I'm kin to these worms?"

"Probably not but I don't know your whole family tree."  Betty's tone indicated she was not interested in my humor.  "Anyway, these worms are on a rampage.  They multiple like rabbits and can move through a 180-acre corn field in about two weeks.  So far only a couple of counties in northern Iowa are badly infected.  Recently, we've had reports that the nematodes are present in Illinois, Minnesota, and South Dakota so they're spreading rapidly."

Not really understanding how my expertise could help in this area, I asked,  "What do they do?  Eat all the kernels and leave just the corncob?"

"Much worse than that.  They don't eat much of the corn but they lay their eggs between the kernels.  Then as they crawl away to die, they secrete an alkaloid that deters other nematodes from laying eggs in the same place.  When the little ones hatch, they eat from the bottom up until they reach the alkaloid.  Then they move to another stalk and start the process all over again.  Only a little of each ear is eaten."

"Sounds like a possible solution to the problem.  Identify the alkaloid and make enough to spray the entire field.  The worms won't have anyplace to lay their eggs.  They probably won't even enter the field."

"If that were the case, I wouldn't need your help,"  Betty hissed, her words as cold as ice.  "The first farmers to be infected didn't realize their corn contained these nematodes or this novel compound.  The worms are quite small and can be seen only under a microscope.  When the farmers fed the corn to their cattle, the cows went crazy.  Something like an epileptic seizure.  They all died within 48 hours of exposure.  All the other farms animals, chickens, pigs, turkeys, are also affected and quickly die when exposed.  We even have one report of a farm boy dying after eating eggs from contaminated chickens.  However, other eggs collected at the same time did not cause any adverse effects so this death is probably not related to the alkaloid.  At least we hope not.  So far the press hasn't picked up on this problem but when they do, you can imagine what they'll print."

I pulled on my beard, a habit I had when thinking.  "I can also picture various groups demonstrating and wanting the government to do something, anything.  That's what these groups usually do when something threatens to upset their comfortable life style."  I paused when I heard Betty sigh, indicating she was not interested in this type of discourse.  I continued, "That is some toxin!  What's being done to prevent these worms from spreading?"

"The USDA is tracking their movement.  Any field found to be infected is cut down and the crop burned.  The fields next to it are closely monitored and when,"  she stopped for a second.  "Note that I said when and not if.  When these fields become infected they are also burned.  Even cutting and burning the corn in a couple hundred yards circle around an infected field doesn't stop the nematodes.  Somehow they move through the cleared area and into another field."

"That's very interesting, Betty."  I hoped my next question would lead to some real consulting business.  "How can I help you with this nematode problem?"

Betty replied in a somewhat harsh voice,  "Same old thing.  Some of our scientists think they've discovered the wonder drug.  They've sold management on their theory and want to rush into large field testing with only a small amount of laboratory data.  Sound familiar?"

"Sure does."  I smiled, my lips curling in a devilish grin.  Now she was talking my language.  "When someone tells me that they have a wonder drug, I wonder if it will ever become a drug."  More seriously I asked,  "Are they hoping to treat the nematodes with this compound?"

"Yes they are.  I'd like you to come up here for a couple of days, David, and help me convince the company leaders that we need to conduct carefully controlled studies to evaluate this compound.  We still have the confidentially agreement in place and we'll pay you your normal consulting fee and expenses."

I shook a clenched fist at the ceiling.  At least two days of work, my mind shouted happily.  Maybe more.  Thank you, Betty.  Struggling to keep my voice calm, I said,  "No problem.  When do you want me?  Right now my calendar is fairly open."

I detected some relief in her voice.  "If you catch an early flight tomorrow, we can start the discussions almost immediately.  We already have a ten thirty meeting scheduled so everyone that needs to be there will be available."  Chuckling, she added,  "That will be most of our scientific staff.  I'll pick you up at the airport and we can talk some more on the way in."

"See you soon.  I'm looking forward to working with you, Betty."  I hung up and stared out the window at the white, fluffy clouds racing across the late afternoon sky.  I thanked my lucky stars for Betty and her nematode problem.  After checking the e-mail note Betty had sent and not learning anything more, I quickly made the necessary arrangements for the trip.  As I tossed a couple of unread scientific journals into my briefcase, I considered,  This meeting must be very important.  Must be something major.  Wonder why she waited so long to call me if her company needs my expertise so urgently.  Guess I'll find out tomorrow.  I left the office a happy man who again had a purpose in life, at least a purpose for the next couple of days.

Stepping out of the terminal and feeling the cold wind hit my face brought back memories of my days at Hansen.  Snow would be falling soon and the poor souls living here would be digging out their driveways so they could go to work.  In many ways I envied them, at least the part about having to go to work.  Not wanting to miss Betty, I glanced into each passing car.  We had not seen each other for a couple of years and I had grown a beard and put on a few pounds.  Of course, the extra weight was in all the wrong places.  After a few moments, my mind started to wander, mostly on my lack of work and financial problems.

"David,"  a voice called from across the five-lane airport arrival road,  "David, over here."

I shook my head and peered in the direction of the sound.  Smiling broadly when I saw Betty, I waved and shouted,  "I see you, Betty."  I picked up my briefcase and overnight bag and worked my way through the many parked cars.  "Betty dear,"  I encircled her thin frame with my arms.  "I'm so glad to see you.  It's been so long.  Too long."

Betty returned my hug and then leaned back slightly, her baby blue eyes shining up at me.  "David, you haven't changed a bit.  You're still as handsome as ever."

I knew she was lying but appreciated the comment.  Friends say the nicest things.  "I wish that were true.  How do you stay so lovely?  And so slim?  Wish I had your metabolism rate!"

Betty laughed, the laugh I had missed hearing for the past few years.  "That's why I like you, David.  You always have lied well.  Let's get going.  We have a meeting to attend."  She broke away from me. 

After throwing my stuff in the back seat, I climbed into the front.  "Anything more you can tell me, Betty?  Why is your company trying to find a new compound to control these nematodes?  Don't normal pesticides kill the worms?"

Before answering, Betty started the car and moved into traffic, quickly picking up speed.  "None of the available pesticides or nematicides are effective against the nematodes.  Only banned ones, like DDT, seem to work and then not completely.  Even with a kill rate of over 99%, enough nematodes are left to infest the next field."

"How are these nematodes controlled in their natural habitat?"  I still wondered how my drug development expertise could help her with the problem.  "Something must be preventing them from killing all the animals in Asia."

"The ornamental grass only grows in a relatively remote area on a high plateau."  Betty glanced at me, her eyes showing her unhappiness.  "Not many animals around.  The nematodes are controlled by spiders that sting them with a poison.  The venom contains both neurotoxins and hematoxins."

"These spiders aren't affected by the alkaloid?"

"We don't think so.  We considered introducing the spiders to control the nematodes."  Betty shivered slightly, displaying her dislike of spiders.  "However, that might be like throwing gasoline on a burning fire.  Once the spiders took care of the nematodes, they could become a pest themselves since we don't know how they are controlled in their normal environment.  We might have to bring another species to control the spiders."

Chuckling, I said,  "Reminds me of an old campfire song about the old woman who swallowed a fly."  Betty smiled but her eyes told her she was not in the mood for my humor.  I continued more seriously,  "This is some situation.  What does KornBiotech have to do with it?"

"Like a few other agrobiotech firms, we are heavily involved in seed corn production and are attempting to genetically engineer the corn to give higher yields and to be more resistant to disease and parasites."  With pride in her voice, she said,  "My staff and I have found a number of enzymes that we can regulate so that the kernels produced are larger and contain more nutrients.  We have encoded the regulatory proteins into a seed corn strain and are conducting field tests.  No problems yet and we think we can increase the yield by 10 to 15%.  Quite an improvement."  She frowned, small wrinkles displayed in the corners of her eyes.  "Back to the nematode problem.  So far we've lost only a couple of our fields but if enough of the seed corn becomes infected, the farmers won't have anything to plant the next year.  If we don't find a way to stop these nematodes, we might have to stop growing corn, which would devastate the meat producers and cause major problems with the human food supply."

"We do like our meat, don't we!"  I grinned sarcastically.  "Of course we don't want it to cause us to run around in circles."  Again noticing that Betty was not in the mood from my bad jokes, I continued more soberly,  "What are you doing?  Why do you need my help?"

"As you might expect, we're working on multiple possibilities.  My group is trying to find a way to genetically engineer the corn so the worms don't like it.  Another group has been searching for an analogue of the alkaloid that is not toxic and still repels the worms.  That's where you come in.  Our chemistry group synthesized a whole library of analogues.  Almost a hundred thousand unique chemicals.  These were screened by coating ground corn with the compounds and placing the mixtures into 96-well titer plates.  Nematodes were then introduced and allowed to eat whatever corn they wanted.  Only the wells still containing corn were considered to have a positive response."

"A negative response to the nematodes,"  I said so softly that Betty could not hear.

"The five hundred or so positives,"  Betty did not notice my smirking expression,  "were then tested in an in vitro  epilepsy screen that measures the electrical activity of neurons in culture.  Those compounds that did not produce changes in activity were then tested for dose response in both the primary corn-nematode screen and the neuron-epilepsy screen."

"Let me guess,"  I said harshly as the pattern in the discovery process became clear.  "The compound with the most potency against the nematodes was selected.  Did they even consider the results from the epilepsy screen."

Betty smiled and blew me a kiss.  "After I forced them, they did.  I learned that from you."  She paraphrased one of my favorite statements.  "Consider all the available data before making a choice.  If you don't, you'll most likely be surprised and not pleasantly surprised."

"Very true,"  I muttered, pleased that she remembered.

"Their first and second choices, based on nematode activity or rather deterrence, caused electrical activity in neurons at concentrations about three times that needed to stop the worms.  The third most active compound looked good but the chemists killed that one because they thought it would be too difficult to synthesize in quantity.  Something about a sterically hindered side chain."  Betty frowned, suggesting she was attempting to remember long unused chemistry from her undergraduate days.  "The fifth, sixth, and ninth compounds also produced some activity in the epilepsy screen.  So compounds four, seven, eight, and ten were selected for whole animal experimentation."

"So far so good, Betty.  You've been keeping them on track."

"Not any longer, David."  Betty grimaced and her tone showed her displeasure.  "They dosed rats with each of the alkaloid analogues and watched for seizures.  Only two doses were used, one the level that repels the nematodes and one three times higher."

"Why didn't they run a full dose response curve?"  I asked with concern in my voice.  "Did they check for anything other than seizures?"

"They only did the tests because a couple of us thought they were necessary.  Live animal experimentation is expensive.  When compound four didn't cause any seizures in the six rats tested, the pharmacology group wanted to immediately start field testing.  They want to spray a whole corn field with this compound to see if the nematodes will stay away."

"Isn't that a little bit extensive?  Shouldn't some additional, more controlled testing be done first?"

"Now you know why I've asked you to come here."  Betty's hands lightly pounded the steering wheel.  "I need you to help me convince the CEO and VPs that we shouldn't rush into this large field test."  She glanced down at her watch.  "The meeting has already started.  Wish I could tell you more but I think you have some idea of my dilemma."

Betty stopped talking as she flashed her badge and drove through a guard gate.  To our right were a number of modern three story buildings.  Most likely the office space for the bigwigs and the marketing staff of the many small firms that occupied this industrial park,  I considered as Betty drove into the parking lot.  The laboratories are probably in the box-like buildings further down the road.

Betty pulled into a parking space and we got out of the car.  The cold air caused my eyes to water but I breathed deeply, enjoying the crisp, fresh air.  As we hurried up the walk, I summarized,  "You, and the rest of the country, have a problem with these nematodes which may, will affect our ability to feed the people.  Your scientists have found a compound that may prevent the worms from laying their eggs in the corn but the compound has undergone only limited testing.  The proposal is to move from the discovery phase directly into a large field trial without first characterizing the compound for possible demerits."

"That's about it, David."  Betty stomped her foot on the cement.  "Just like some pharmaceutical compounds we worked on.  Go directly from drug discovery to human clinical trials with the minimum of testing necessary to satisfy the regulatory agencies.  The success rate in ag isn't much better than the one-out-of-a-hundred clinical drug candidates that becomes a therapeutic product.  I don't have a good feeling about this compound."  She continued in a harsh, bitter voice,  "About this whole approach to the nematode problem!"

As we entered the plush lobby with large, light brown leather chairs, glass tables covered with magazines and company literature, and numerous large, green plants that almost reached the high ceiling, I thought about how little I knew about agriculture products and how they were tested and developed.  I hoped my pharmaceutical training could help Betty with her problem.  After signing in and sticking a visitor's badge on my suit coat, we waited for the elevator.  I pondered,  Something's not right.  Betty isn't telling me everything.  What is she holding back?  Doesn't really matter.  She's paying me and from what she said, this analogue definitely needs more characterization.  I'll do my best to help her convince her bosses that they should spend a few dollars and a little time now so they don't waste a whole lot of both later.

Betty led me into a darkened conference room.  In the dim light of the slide being projected, I could see a long, dark conference table around which a number of men and women were sitting.  Many appeared to be listening to the speaker who was describing the slide.  Others were making notes or drawing pictures or just staring, apparently bored with the presentation, probably because they had heard it before.  I glanced at the graph being displayed and saw the numerous S-shaped curves that told me the data were on biological activity.  The speaker, standing to one side and not very visible in the dim light, was pointing to one curve and telling the audience why this analogue was superior to the others.  Based on the data displayed, he was right.  The voice speaking caused me to shudder.  It was familiar and not pleasantly so.

We moved past some people sitting along the wall and found a couple of empty chairs.  As we sat down, the speaker concluded,  "So based on our experiments, analogue number four, designated A4, has the highest potency in deterring the nematodes, only very limited activity in the epilepsy screen, and no observable effect in rats.  We recommend that this compound be put on a fast track and that field testing begin as soon as possible."

"Lights please,"  droned a deep voice.  "Thank you, Dr. Bass, for your summary.  Your groups' data is quite convincing.  The meeting is now open for discussion."

The lights came on and I squinted at the tall, thin, dark haired man standing in the front of the room.  Dr. William Bass was also a former Hansen Pharmaceuticals employee but unlike Betty and me, was not one of those fired.  Our paths had crossed many times and usually ended up in shouting matches.  He had been an associate director of the virology group and believed that biological potency was the key, the only key, to successful drug discovery.  From the hateful stare he was directing at me, I knew he was thinking about the lead compound that Betty and I had helped kill because it could not be effectively delivered to the site of action.

Smiling, or rather grinning diabolically, Bass said in his high pitched voice,  "I see Dr. Mills has finally arrived and has brought with her Dr. David Wilcott.  Nice to see you again, Dr. Wilcott." 

I knew he was lying.  My feelings at seeing him again were similar. 

Sensing my tension, Betty gave my arm a slight squeeze.  She stood up.  "Sorry I'm late.  I would like to introduce Dr. David Wilcott.  He is a consultant with expertise in the development of compounds.  I believe he can help us understand what we might need to do in order to ensure that this A4 alkaloid is the best one for further development."

"As I just shared,"  said Bass curtly,  "our data clearly indicates that A4 has good potency and limited toxicity.  What more do we need?"

The game was on and I was hardly prepared.  Other than the limited information Betty had shared, I knew little to nothing about this project.  Slowing standing, I considered my options.  I did not like any of them.  I scanned the faces now turned toward me and noted those that were frowning and probably in Bass' camp.  Most of the others could not be easily read so they were either waiting for me to enlighten them with some words of wisdom or were wishing the meeting would end so they could go to lunch.  I said slowly,  "I am pleased to be with you today.  Betty, Dr. Mills, has provided me with some background information on your project.  These nematodes will become quite a problem unless you can find a way to stop them from wiggling their way through corn field after corn field."  I paused and heard a few chuckles coming from around the table.  "Dr. Bass,"  I nodded toward the scowling man still standing in the front of the room,  "and his group have apparently discovered an analogue of the nematode's alkaloid that has some promise in stopping this infestation."

"Some promise!"  Bass shouted.  Most heads turned toward the outburst.  "We have the agent that's going to save both the farmers and the meat industries.  It's safe and effective.  It will make KornBiotech a major player in the agricultural field."

"I won't argue whether or not your compound is effective, Dr. Bass."  I had not waited for anyone else to answer the comments.  "Your data does suggest that this analogue, A4 I believe you call it, will most likely block the worms from laying their eggs under the corn kernels.  However, I would like to consider a couple of other issues, including safety."

Bass glared hate at me but said nothing.  The leader of the meeting, probably the vice president of discovery or maybe a senior vice president in charge of research and development, said in his deep voice,  "That's why we're here, Dr. Wilcott.  Ask away.  We'll do our best to answer your questions."

I liked the man already.  He was apparently interested in hearing all sides before making a decision.  "The first item deals with the physical chemical properties of A4.  What do you know about the solubility?  Is A4 stable under the conditions to be used for application and in the environment after application?

"I believe I can answer that question."  A gray haired woman stood up and nodded a greeting at me.  "I'm Dr. Stevenson and in charge of chemistry.  First, A4 has only limited solubility in water.  We plan to apply it to the field as an emulsion.  We have used similar technology to spray other relatively insoluble herbicides and pesticides."

I knew that information might be important and filed it in my mind for use later.

She continued,  "We don't appear to have any problems with stability.  So far we've tested A4 to a variety of conditions including temperature, acids and bases, and UV radiation.  No degradation was found.  We did learn that the compound is substantially more soluble in acid than in base.  In fact, above pH 7 A4 has the solubility of sea sand."  A few laughs filled the air as she returned to her seat.

"Thank you, Dr. Stevenson."  I smiled at the lady who returned my smile.  She was not my enemy, like Bass, and might be a friend.  "That answers that question."  I still had a few other issues in this area that needed to be answered but those could wait.  "My second question deals with the rat study that was conducted.  How were the animals dosed?"

Bass cleared his throat, his expression saying he did not want anything to do with me but knowing he had no choice.  "We used our standard procedure.  A4 was dissolved in DMSO and the solution was diluted one to nine with normal saline containing zero point one molar citric acid.  The rats were orally gavaged with the mixture.  We observed them for 48 hours and detected no signs of seizure."  His voice got louder and louder as if he was trying to convince me with noise rather than data.  "These results indicate that A4 will not be toxic to the farm animals who eat the corn."

"I'm not so sure,"  I said, attempting to keep my voice as calm as possible.  "First, the rats were dosed with an acidic solution that we were just told is optimal for solubility.  What happens when A4 reaches the upper GI tract where the pH is higher?  Does it stay in solution or fall out?  If it precipitates, how much is absorbed into the body?"  Before anyone could respond to any of these questions, I continued,  "Second, assuming A4 is absorbed, how much gets by the liver?  Many compounds are rapidly cleared, either by metabolism or excretion into the bile, and never reach their target, in this case the brain."  I saw a couple of groups whispering.  From their facial expressions, I knew at least some were paying attention and understanding my comments.  "Which raises another point.  Do you know that the alkaloid itself is responsible for the toxic effects observed in the farm animals?  Could a metabolite be the problem and not the parent compound?  Since the alkaloid and A4 are structurally similar, could they produce similar metabolites, one or more of which may cause the observed toxicity?  And finally, at least for now, do you know if whatever causes the seizures can penetrate the blood brain barrier of rats?  Different species can have completely different delivery, metabolism, and toxic effects to the same compound.  In my opinion, the rat study does not show that A4 is safe to farm animals."

Betty smiled up at me.  She rose from her chair.  "Thank you, David.  Your insight into this issue is why I wanted you here.  You've raised some important points, any of which could cause A4 to have a safety issue.  What do you suggest we do?"

Before I could answer, Bass shouted, his loud voice filling the small conference room,  "We don't have time or the resources for any extended, costly studies.  The nematodes will be all over Iowa next year.  We have to be ready to start spraying in early spring to keep the worms out of newly planted fields.  If we don't stop them, they'll be in all the corn the year after, including the seed corn."  His voice rose to new levels.  "All the cattle, pigs, chickens will starve."

"I'm afraid he's right, Dr. Wilcott,"  the leader said in a relatively calm voice.  "We don't have the luxury of time and can't possibly obtain the data you,"  he nodded at Betty,  "and a few others, think is necessary.  We have to act and act quickly.  Based on what Dr. Bass told us earlier, I think we should go ahead with the field test.  Are there any objections?"

Bass beamed at his apparent victory.  Betty sat down hard, her face buried in her hands to hide her defeated expression.

"Sir?"  I asked quietly.  The leader glanced at me and nodded.  "Before you decide, I'd like to make at least one more point.  Earlier, Dr. Stevenson said A4 was very stable.  In fact, she indicated that her group had not been able to degrade the compound under various test conditions."

"That's right!"  Bass shouted happily.  "A4 will have to be applied only once and will stay effective for the whole growing season.  The farmers will love it."

"Just my point, Dr. Bass,"  I said more sharply than I intended.  That arrogant fool was getting to me.  Taking a deep breath to calm myself, I continued,  "Let's assume you conduct your field test and the results are all thumbs up.  The nematodes are prevented from infecting the field."

"As will be the case!"  exclaimed Bass from the podium.  "We have a guaranteed winner here."

"Let him finish, William,"  the leader responded, his strong voice telling Bass who was in charge.  "I want to hear his point even if you apparently do not."

Noting the surly expression on Bass' face from the rebuke, I inwardly smiled, struggling to keep my feelings from being displayed.  I had enough enemies and did not need to make more.  Bass probably had a number of friends in the room.  Someday they might be a client or know someone who needed the services of a consultant.  If I was to stay in business, I needed all the recommendations I could get.  I carefully chose my words,  "Where does this stable alkaloid go?  My guess, based on Dr. Stevenson's comments on its solubility and stability, would be that it will stay on the crop.  What happens when the corn is eaten by the farm animals?  Do they get sick and die?  Do they store the alkaloid in their meat to be passed on to humans?  Do humans process the chemical differently than the farm animals?"  I could see that Bass was ready to respond.  Before he could state his thoughts, I said quickly,  "From the present data, you really don't know, do you?  You only have one live animal study and that one only provides limited results on the safety of A4."

"I see your point, Dr. Wilcott,"  the leader said, his eyes narrow in thought.  "If A4 ends up in humans and causes only a few to get sick, we would be put out of business.  The lawyers would eat us alive."  I heard a few controlled laughs.  "We're damned if we don't move forward and may be damned if we do.  Any ideas?"

"A4 is safe,"  screamed Bass, his face distorted in anger.  "I'd bet my reputation on it!"

I felt Betty chuckle and knew she was thinking that the former virology head did not have much of a reputation to lose.  "I have a couple of ideas that may help you out of your dilemma."  I hoped the leader would listen and then allow me to implement the program.  The consulting fee could run into weeks.

"Wilcott, your ideas always cost too much money and take too long."  Bass' voice was filled with hate.  "All you've ever done is kill good compounds.  You've never been anything but a devil in disguise.  I should know.  I've felt your forked tongue often enough."

After glaring at Bass to keep quiet, the leader said,  "Continue, Dr. Wilcott.  What do you suggest?"

"What I think you need is a plan to more fully characterize A4."  I noted that most of the people in the room were now paying close attention.  "This plan should include a limited field test to ensure that A4 prevents the nematodes from infesting the corn.  While this test is being set up, I recommend that a concentrated mixture of A4 be sprayed onto corn and the amount present determined by an analytical chemistry method that I'm sure Dr. Stevenson has available."  I smiled at the scientist who nodded that I was correct.  "The corn can then be ground and mixed with normal corn to give various A4 dose levels.  This spiked corn can be fed to rats and at least one farm animal, say chickens or pigs.  Since adverse effects to the nematode's alkaloid occur shortly after the corn is eaten, this test can be quite short, maybe only a few days of dosing.  The results will provide you with a much greater indication of safety, particularly since the test animals will be exposed to A4 in a manner similar to farm animals."

"That's just an acute toxicity study.  No problem there."  A heavy set, balding man smiled at me.  "We can also check all the organ systems for signs of toxicity."

"Anything else?"  The leader nodded his agreement to the scientist.

"Just a couple more thoughts."  I glanced down at Betty, her blue eyes again shining.  "You might want to consider running an in vitro metabolism screen using hepatocytes from various species including humans.  That way you'd have an indication if A4 is metabolized and if all the species handle A4 the same way."

"We don't have the capability to do that!"  said Bass harshly but quietly.  He did not want the leader to be anymore upset with him than he already was.  "That's a specialized test that is done only at the major pharmaceutical houses.  We're just a small firm and don't have much metabolism expertise."

I had expected that response.  I smiled  at Bass for giving me the opening.  "Not any longer.  A number of CROs now offer that as one of their services.  The cost is reasonable and the turn around time quite short.  You should be able to have results in a couple of weeks."

"My group can handle that."  A young, dark haired woman glared at Bass.  She was apparently the metabolism expert and did not appreciate the comment on her abilities.  I had another friend in the room.  "We only need Dr. Wilcott to give us the names and contact information for these contract research organizations."

The leader smiled his thanks.  "Anything else?"  His tone suggested that he hoped I was done.

"Yes,"  I said, wondering if I was pushing my luck.  "I suggest that you adapt the analytical chemistry method to a bioanalytical method.  You apparently already know how to extract A4 from corn so a similar procedure should work for plasma.  Another group of animals can be dosed and serial blood samples collected and analyzed.  The results will provide information on the extent of absorption and how fast A4 is cleared from the body.  If you want to gain more information on potential differences between species, you could collect tissues at say 24 hours after dosing and determine if any organ has high concentrations of A4."

Dr. Stevenson said,  "We can provide the assays if toxicology can dose the animals and collect the samples."

The heavy set, balding man, apparently in charge of the animal group, answered,  "We'll work with you.  You just tell us when you want us to bleed the critters."

"Anything else?"  the leader asked somewhat curtly.

I took a deep breath, knowing he and the rest of the people in the room, especially Bass, would probably not like my last suggestion.  "I do have one more thought for you to consider.  Everything we've talked about so far has been on A4, which is your lead candidate."  I noted a number of nodding heads.  Even Bass kept his mouth closed, however difficult that was for him.  "Dr. Mills told me earlier that you have three other analogues that showed some promise in your nematode and epilepsy screens."

"That's right,"  said the leader.  "They have less potency than A4 and don't show any neuronal activity.  What's your point?"

I quickly glanced around the room.  Only Bass was frowning at me.  "Right now you have all your eggs in the A4 basket.  If A4 fails for any one of several possible reasons, you are back to square one.  Since time is of the essence and you need to select the best compound, I suggest you test the other three compounds along with A4.  And that you use the nematode alkaloid as a positive control in your studies."

Bass could no longer hold back.  His face red with anger, he almost screamed,  "That's five times the work!  Five times the time!  Five times the cost!  Do you think we're made of money, Wilcott?"

"No I don't, Dr. Bass,"  I said as calmly as possible.  "However, if you want to have a candidate ready for spraying next year, you might want to consider expending the resources to get the data, which with appropriate planning can be obtained simultaneously.  If the overall project is as important as I think it is, can you afford to take the risk of studying a possible looser while a winner sits on the shelf?  If all the results are available, you can select the optimal compound, one that is both effective and safe."

The leader glanced down at his watch.  "I think this meeting has gone on long enough.  It's way past lunch time.  I suggest we break for an hour and then reconvene to discuss what Dr. Wilcott has recommended."

As the others pushed away from the table and left the room in small groups, where a number of heated discussions were taking place, Betty stood up and smiled, her face beaming in apparent victory.  "I knew you could convince them, David.  They'd never listen to me.  They think I'm just a molecular biologist."

"Not all of us, Betty."  The leader joined us.  "I want to thank you, Dr. Wilcott, for your comments and suggestions.  You've given us a lot to think about."

"My pleasure, Dr. ...",  I paused and glanced at Betty.  "I don't believe we've been introduced."

"Oh, I'm sorry."  Betty's face turned slightly red.  "David, this is Mr. John Bass.  He's the president and CEO of KornBiotech."

I tentatively held out my hand, hoping he would not bite it off.  "Nice to meet you, Mr. Bass.  Are you related to William?"

"He's my nephew."  Mr. Bass took my hand in a strong grip.  I returned the pressure and shook his hand.  "My brother asked me to hire him after his problems at Hansen.  Sometimes I wish I hadn't.  He can be such a pain at times."

Sighing deeply with what I knew was relief written all over my face, I stammered,  "He's probably a very good scientist in his field of expertise."

"You don't have to make amends with me."  Mr. Bass smiled.  "I know what he is.  Now if you will excuse me, I want to grab a bite to eat before the meeting continues.  It was nice meeting you, Dr. Wilcott.  We'll call you when we need you."  He quickly left the room.

Chest fallen at the dismissal when I had hoped for a much bigger role, and consulting fee, in the design and interpretation of the studies I had recommended, I attempted unsuccessfully to smile at my friend.  "Guess that's that.  Now what, Betty?  Mr. Bass apparently thinks his team can do the job without any more of my input."

"You did great, David."  Betty gave my arm a slight squeeze.  "Better than I ever hoped."  She started for the door.  "I'll give you a ride to the hotel.  You can catch a flight home early tomorrow morning.  Tonight you should celebrate on KornBiotech."

"Would you like to join me for dinner?"  We walked down the corridor to the elevators.  "We could discuss old times or even new ones."  I needed some company to overcome my growing disappointment and depression.

"I won't have time, David."  Betty stopped and gazed up at me, her eyes large with excitement.  "Now that you've given Mr. Bass a program that will either kill all the analogues or provide one with an acceptable chance of success, I have to get my own team started on our experiments." 

She paused and I knew she was considering whether to tell me more.  She did have another agenda,  I mused as I waited.  She wanted me to kill the analogue program so her own idea would be more acceptable.

After a moment, she continued, her voice displaying her happiness.  "I probably shouldn't tell you this but I'm going to anyway.  My team is working to genetically engineer corn so the nematode enzymes for production of the alkaloid are expressed and can be used to generate a nontoxic analogue.  We'll use the corn itself to stop the nematodes."

I frowned at her comments, knowing my friend was not fully aware of the problems she might cause.  "How do you know the produced alkaloid won't be toxic?  How will you control the rate of synthesis?"

Her eyes glared at me.  "Don't start on me, David,"  she said, her voice harsher than I had ever heard before.  "We know what we're doing.  We can manipulate the code to produce whatever compound we want and can control the amount of production.  The data from the studies you recommended will tell us which alkaloid is best.  We'll grow genetically engineered seed corn in green houses this winter and be ready for planting next spring."  She paused and grinned diabolically, turning her normally pretty face into a fiendish mask.  "I'll be famous.  My corn will solve this problem."

I sighed deeply, knowing that while I might have prevented one poorly designed program from moving forward too quickly, I had allowed another, equally problematic project to move ahead unchecked, somewhat like the nematodes themselves.  Hopefully a number of other agriculture firms were also working on this problem and would be able to find a solution.  Somehow I doubted that Betty's firm was going to be successful.  "Take care your experiments, Betty,"  I said unhappily as I entered the elevator.  "I'll catch a cab.  Nice seeing you again."

As the doors closed, I heard her say,  "Thanks, David.  Thank you so much."

I sat at my desk, hoping the phone would ring.  The smoke from my pipe curled toward the ceiling.  I took a last draw and put it down, vowing once again to quit smoking but knowing I would probably not be successful. Outside, the early spring sun was shining brightly and nature was awakening with some early flowers already in bloom and trees starting to bud.  Business had not been good over the winter.  Only a couple of days of consulting revenue, but that is another story, and two commissioned reports that paid a small honorarium.  Just enough to keep the lights on and the creditors off my back.  I glanced at the words displayed on the computer screen.  I was preparing a marketing letter that offered pharmaceutical and biotechnology companies a training course in the development and validation of bioanalytical chemistry methods.  Maybe a few of the executives on my mailing list would read the letter and hopefully one or two would recognize the opportunity being offered and contract with me to present the short course.

Betty called last week.  She was crying and I knew she was upset.  I wanted to give her whatever comfort I could.  She is a dear friend, even if she had used me in an attempt to advance her career.  Through sobs, she told me that KornBiotech had conducted some of the experiments I had recommended but had studied only A4 and A7.  Both compounds showed extensive metabolism by hepatocytes from all the species tested, were toxic in rats when dosed on corn, and caused seizures in chickens.  The primary toxin was one of the metabolites but they did not know which one.  Since they had not studied the other two analogues, they did not have a candidate ready for spraying.  Betty told me her group had engineered seed corn to produce both analogues.  When she learned a metabolite was the cause of toxicity, she was devastated, all her work for nothing.  She told me Mr. Bass was shutting down the project since another small agrobiotech firm had discovered an alkaloid analogue, not substantially different from their A10, that was not metabolized to a toxic substance and that had sufficient potency to deter the nematodes from laying their eggs.  This firm's field tests were all positive and the submission for governmental approval made.  Farmers were anxiously awaiting the soon-to-be-approved product to protect their corn crops.  The nematode infestation problem was apparently solved or at least under control.  KornBiotech had not been the winner.

Shaking my head to clear my thoughts, I started to type.  I needed to get the marketing letter out this week.  I needed the business.