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Authors: Robin Cook

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“What about this hooligan who attacked you? Don't we have to worry about him coming back?”

“That's a concern but apparently a risk we have to take. He's not from the Bahamas, and my educated guess is that he's already on his way home. He said that if he had to fly the hell back down here from Boston again, he'd, and I quote, hurt me bad, which leads me to believe that New England is his usual hangout. At the same time, he said he didn't want to hurt me so bad that I couldn't get the company back on its feet, meaning they have a vested interest in my well-being, despite how I feel at the moment. But most importantly, I'm hoping your phone conversations with your mother, which
will undoubtedly get communicated to your brother, will convince the Castiglianos it's worth waiting three weeks.”

“Should we change hotels, since I told my mother we're staying here?”

“I thought about that while I was sitting in the car, waiting for you to come out of the store. I even thought about taking Paul up on his offer to stay out at the Wingate Clinic.”

“Oh, God! That would be like going from the frying pan into the fire.”

“I wouldn't want to stay there either. It's going to be bad enough putting up with those charlatans during the day. So I think we should just stay here, unless it's going to drive you crazy. I don't want a repeat of our night in Turin. My feeling is that we should stay put but not leave the hotel, except to go to the Wingate Clinic, which, starting tomorrow, is where we are going to be most of the time anyway. Agreed?”

Stephanie nodded a few times as she absorbed everything Daniel had said.

“Do you agree or what?” Daniel asked. “You're not saying anything.”

Stephanie suddenly threw up her hands in a burst of emotional frustration. “Gosh, I don't know what to think. You getting attacked just adds to my uneasiness about this whole Butler affair. From day one, we've been forced to make assumptions about people we know little or nothing about.”

“Wait just a second!” Daniel growled. His face, already red, got redder still, and his voice, which had started out low, began to rise progressively. “We're not starting the debate again about whether or not we're going to treat Butler. That's been decided. Our current conversation is about logistics from this point on, period!”

“Okay, okay!” Stephanie said. She reached out and put a hand on his arm. “Calm down! Fine! We'll stay here and hope things work out for the best.”

Daniel took a few deep breaths before saying, “I also think we should make it a point to stay together.”

“What are you talking about?”

“I don't think it was an accident the muscleman assaulted me when I happened to be alone. Your brother obviously doesn't want you hurt; otherwise, we both would have been
slapped around, or at a minimum, I still would have borne the brunt, but you would have had to witness it. I think the man waited until I was by myself; ergo, I believe our staying together at all times away from our room would provide a certain amount of safety.”

“Maybe you're right,” Stephanie mumbled equivocally. Her mind was a jumble. On the one hand, she was relieved that Daniel wasn't making a negative reference to their relationship when he mentioned staying together, while on the other hand, it was still hard for her to admit to herself that her brother could have had anything to do with the violence Daniel had experienced.

“Can you get me some more ice?” Daniel asked. “What I've got is just about melted.”

“Of course,” Stephanie said. She was relieved to have something to do. She took the soggy towel and exchanged it for a fresh one in the bathroom. Then she revisited the ice bucket on the bar. When she handed the fresh ice pack to Daniel, the phone on the side table suddenly sprang to life. For a few moments, its repetitive jangle inundated the otherwise silent room. Neither Daniel nor Stephanie moved. Both stared at the phone.

“Now, who the hell could that be?” Daniel questioned, after the fourth ring. He positioned the ice pack on his eye.

“Not very many people know we are here,” Stephanie said. “Should I answer it?”

“I suppose,” Daniel said. “If it is your mother or brother, remember what I said earlier.”

“What if it's the person who attacked you?”

“That's highly unlikely. Answer it, but be nonchalant! If it is the thug, just hang up. Don't try to engage him in any conversation.”

Stephanie went to the phone, picked it up, and tried to say hello normally while looking back at Daniel. Daniel watched her eyebrows raise slightly as she listened. After a few moments, Daniel mouthed, “Who is it?” Stephanie held up her hand and motioned for him to wait. Finally, she said, “Wonderful! And thank you.” Then she listened again. Absently, she twirled the phone cord with her finger. After a pause, she said, “That's very nice of you, but it's not possible tonight. In fact,
it's not possible any night.” She then said goodbye in a clipped tone and replaced the receiver. She returned her eyes to Daniel's but for a moment didn't speak.

“Well? Who was it?” Daniel demanded. His curiosity was getting the best of him.

“It was Spencer Wingate.” Stephanie shook her head in amazement.

“What did he want?”

“He wanted to let us know that he located our FedEx package, and he's arranged to have it delivered first thing in the morning.”

“Hooray for small favors. That means we can start creating Butler's treatment cells. But that was a rather long conversation for such a short message. What else did he want?”

Stephanie gave a mirthless laugh. “He wanted to know if I would come to his house in Lyford Cay Marina for dinner. Strangely enough, he made it clear that the invitation was just for me and not for us as a couple. I can't believe it. It was like he was trying to hustle me.”

“Well, let's look on the bright side; at least he has good taste.”

“I'm not amused,” Stephanie countered.

“I can see that,” Daniel said. “But let's keep the big picture in mind.”

eighteen

11:30
A
.
M
., Monday, March 11, 2002

 

Occasionally, Daniel had
to give credit where credit was due. There was no doubt in his mind that Stephanie was far better at cellular manipulation than he, and that reality was underlined by what he was presently watching through the eyepieces of a double-headed dissecting stereomicroscope. He and Stephanie had placed the instrument on the corner of their lab bench at the Wingate Clinic to allow Daniel to watch while Stephanie worked. Stephanie was about to begin the process of nuclear transfer, otherwise known as therapeutic cloning, by extracting the nucleus of a mature oocyte whose DNA had been stained with a fluorescent dye. She already had the human egg cell fixated by suction with a blunt-tipped holding pipette.

“You make this look so easy,” Daniel remarked.

“It is,” Stephanie responded, as she guided a second pipette into the microscopic field with a micromanipulator. In contrast to the holding pipette, this pipette's hollow end was as sharp as the finest needle, and the pipette itself was only twenty-five-millionths of a meter in diameter.

“Maybe it's easy for you, but it's not for me.”

“The trick is not to rush things. Everything has to be slow and even, and not jerky.”

True to her word, the sharp pipette moved smoothly yet decisively toward the fixated oocyte to push against the cell's outer layer without penetrating it.

“This is the part I invariably screw up,” Daniel said. “Half the time, I go clear through the cell and out the other side.”

“Maybe because you are too eager, and therefore, a bit heavy-handed,” Stephanie suggested. “Once the cell is adequately indented, it just takes a slight tap with the index finger on the top of the micromanipulator.”

“You don't use the micromanipulator itself to do the puncture?”

“Never.”

Stephanie carried out the maneuver with her index finger, and within the microscopic field, the pipette was seen to enter cleanly the cytoplasm of the hapless egg cell.

“Well, you live and learn,” Daniel said. “It proves I'm just a rank amateur in this arena.”

Stephanie pulled away from her eyepieces to glance at Daniel. It wasn't like him to be self-deprecating. “Don't be so hard on yourself. This is busywork, which you've always had skilled technicians to do. I learned how to do it when I was a graduate-student grunt.”

“I suppose,” Daniel said without looking up.

Stephanie shrugged and directed her eyes back into the microscope. “Now I use the micromanipulator to approach the fluorescing DNA,” she said. The tip of the pipette approached its target, and when Stephanie applied a tiny amount of suction, the DNA disappeared up into the pipette's lumen as if the pipette were a miniature vacuum cleaner.

“I'm not good at this part either,” Daniel said. “I think I suck up too much cytoplasm.”

“It's important to get just the DNA,” Stephanie said.

“Every time I watch this technique, I'm even more amazed that it works,” Daniel commented. “My mental image of the submicroscopic internal structure of a living cell is akin to a miniature glass house. How can it be that we can tear out the nucleus by its roots, essentially throw in another nucleus from
an adult differentiated cell, and have the whole thing work? It boggles the imagination.”

“Not only work, but cause the adult nucleus we toss in to become young again.”

“That too,” Daniel agreed. “I tell you, the process of nuclear transfer truly defies belief.”

“I couldn't agree more,” Stephanie said. “For me, the improbability of it working is evidence of God's involvement in the process, which rattles my agnosticism even more than what we learned about the Shroud of Turin.” While she spoke, she guided a third pipette into the microscopic field. This pipette had within its lumen a single fibroblast cell from Ashley Butler's fibroblast culture: a cell whose ancestral nucleus Daniel had painstakingly manipulated, first with HTSR, to replace those genes responsible for the senator's Parkinson's disease with those derived from the shroud's blood, and second, with an added gene at Stephanie's suggestion for a special surface antigen. This fibroblast's nuclear DNA was going to replace the DNA Stephanie had removed from the egg cell.

As Daniel watched Stephanie's artful manipulations, he marveled at what he and she had been able to accomplish in the week and a half since his assault by the thug from Boston. Luckily, his physical injuries had healed and were for the most part a mere memory, save for some residual tenderness along his right cheekbone and the now yellow-and-green remainder of his resolving shiner. Unfortunately, Daniel still struggled with the psychological damage. Burned into the retina of his mind and appearing in recurrent nightmares was an image looming over him of the hulking attacker's huge head, small ears, and bulbous features. Most disturbing was the man's crooked smile and cruel, beady eyes. Even after eleven days, Daniel still suffered repetitive nightmares of that awful face and the feeling of utter defenseless vulnerability it engendered.

In the daytime, Daniel had fared considerably better than during sleep. As he and Stephanie had discussed immediately after the episode, they had made it a point to stay together practically like Siamese twins and not leave the hotel grounds, except to go to the Wingate Clinic. As it turned out, such a
plan was hardly an imposition, since they had spent sunup to after sundown in the laboratory each and every day. There, Megan Finnigan was most helpful, providing them with a small office in addition to their own laboratory bench. Having room to spread out their paperwork and flow sheets was a godsend and a boon to their efficiency. Even Paul Saunders had helped by acting true to his word and producing ten fresh human oocytes twelve hours after they had been requested.

At first, there had been a convenient division of labor between Daniel and Stephanie. Her job initially was to work with the fibroblast culture sent by Peter. She got it thawed and growing with only minor glitches. Concurrently, Daniel attacked the buffered solution containing the shroud sample. After a single pass through the PCR machine to magnify the DNA present in the fluid, Daniel determined the contained DNA was primate and probably human, although decidedly fragmented, as he had expected.

Following a purification trick using microscopic glass beads, Daniel ran the isolated shroud DNA fragments through the PCR several more times before utilizing his dopaminergic gene probes. He was immediately successful, but with only parts of the required genes, a situation that required sequencing the gaps. After several sixteen-hour days, Daniel succeeded in attaching the appropriate fragments with nucleotide ligases to form the genes. At that point, he was ready for Ashley Butler's fibroblasts, which by then Stephanie conveniently had available.

HTSR was the next step, and it went practically without a hitch. Having developed the procedure, Daniel was intimately aware of its subtleties and pitfalls, but under his sure hand, the enzymes and viral vectors worked perfectly, and he soon had a number of the fibroblasts ready. The only problem had been Paul Saunders, who had insisted on shadowing Daniel's every move and frequently got in the way. Paul unabashedly admitted that he planned to add the technique to the Wingate's stem-cell therapy regimen, with the idea of charging the patients significantly more. Daniel doggedly tried to ignore him and bit his tongue to keep from ordering the quack out of his own laboratory, but it was difficult.

Once the HTSR had been completed, Daniel thought they
were ready to do the nuclear transfer, but Stephanie had surprised him with the suggestion that they also transfect the HTSR-altered cell with an ecdysone construct, meaning several combined genes, capable of creating a unique nonhuman surface antigen on the ultimate treatment cells. Stephanie had argued that if there was ever a need or an interest to visualize the treatment cells within Butler's brain after the implant, it could be done with ease, since the treatment cells would have an antigen that none of Butler's other trillion cells had. Daniel had been impressed with the idea and had agreed to the additional step, especially after Stephanie told him she'd had the foresight to ask Peter to send the construct and its viral vector down from their Cambridge laboratory along with the Butler tissue culture. Daniel and Stephanie had used the same technique when they'd successfully treated the mice afflicted by Parkinson's, and it had been a valuable addition to the protocol.

“I always use the micromanipulator for this step,” Stephanie said, pulling Daniel back from his musings. The pipette containing Butler's altered fibroblast pierced the oocyte's envelope without piercing the underlying cell membrane.

“I have trouble with this part too,” Daniel admitted. He watched as Stephanie injected the relatively tiny fibroblast into the space between the egg's cell membrane and its comparatively thick outer covering. The pipette then disappeared from view.

“The trick is to approach the oocyte's envelope tangentially,” Stephanie said. “Otherwise, you can inadvertently enter the cell.”

“That makes sense.”

“Well, I'd say that looks just dandy,” Stephanie said, after viewing her handiwork. The appropriately granular enucleated egg cell and the comparatively tiny fibroblast were locked in an intimate embrace within the oocyte's envelope. “Time for the fusion process and then the activation.”

Stephanie pulled away from the microscope's eyepieces and extracted the petri dish from beneath the microscope's objective. Slipping off her stool, she walked over to the fusion chamber, where she would subject the paired cells to a brief shock of electricity to fuse them.

Daniel watched her go. Along with the recurrent nightmares subsequent to his beating by the Castiglianos' henchman, Daniel struggled with other psychological sequelae from the experience. During the first few days, he had experienced continuous anxiety and fear that the man would reappear, despite what Daniel had reassuringly told Stephanie immediately after the event. It was also despite what the hotel did after Daniel had informed the administration of what had happened. To his credit, the hotel manager had voluntarily stationed a security person within Daniel and Stephanie's building for a week. Every night, the man had accompanied Daniel and Stephanie back to their room after they'd finished their dinner in the hotel's Courtyard Terrace restaurant, and the intimidatingly large individual had remained on guard in the hall until Daniel and Stephanie departed for the Wingate Clinic in the morning.

As Daniel's fear abated during the passing days, his anger at the event waxed, and a significant amount of the anger was redirected toward Stephanie. Although she had apologized and had been sincerely sympathetic initially, Daniel fumed at her lingering doubt about her family's role in the event. She hadn't said as much directly, but Daniel had gotten that sense from indirect comments. With such a screwed-up family and lack of judgment in dealing with them, Daniel couldn't help but question whether Stephanie would be too much of a liability over the long haul.

Stephanie's self-righteousness was also a problem. Even though she'd promised not to make waves with the Wingate people, she was constantly doing so with inappropriate comments about their supposed stem-cell therapy and even inappropriate questioning of the young, pregnant Bahamian women who worked at the clinic, which was an extremely sensitive issue with Paul Saunders. On top of that, she was embarrassingly dismissive of Spencer Wingate. Daniel recognized that the man was being progressively forward in expressing his social interest in Stephanie, a fact that might have been influenced by Daniel's passivity in the face of Spencer's comments, yet there were less rude ways for her to handle the situation than she was choosing. It irked Daniel to no end that Stephanie just couldn't seem to understand that her behavior
was potentially jeopardizing everything. If she and Daniel got kicked out, all bets were off.

Daniel sighed as he watched Stephanie work. Although he felt conflicted over her long-term contribution, there was no question that she was needed in the short term. There were only eleven days left before Ashley Butler's arrival on the island, and in that time, they had to develop the dopamine-producing neurons from the senator's fibroblasts to treat the man. They were making progress with the HTSR and the nuclear transfer already done, but there was a long way to go. Stephanie's expertise with cellular manipulation was sorely needed, and there just wasn't time to replace her.

 

Stephanie could feel Daniel's eyes on her back. She recognized that her sense of guilt and her confusion about the implications of her family's role in his being attacked made her acutely sensitive, yet he was not acting like himself. She could only guess what it must have been like getting beaten up, but she had expected him to recover more quickly. Instead, he was still acting distant from her in many subtle ways, and although they continued to sleep in the same bed, there had been no intimacy whatsoever. Such behavior raised an old concern of hers that Daniel was either incapable or unmotivated to offer the kind of emotional support she felt she needed, particularly in periods of stress, no matter what the cause or whose fault it was.

Stephanie had followed Daniel's suggestions to the letter, so that couldn't be the explanation for his behavior. Despite an aching urge to call and confront her brother, she didn't. And on the relatively frequent conversations she had with her mother, she made it a point to stress that she and Daniel were in Nassau to work, and they were working very hard, which was certainly true. To back it up, she said they had not gone to the beach to swim even once, which was also true. In addition, on multiple occasions she had emphasized that they would be finished soon and would come home about March twenty-fifth to a financially stable company. She had studiously avoided bringing up the subject of her brother with her mother, although on a call the previous day, she had finally yielded to temptation. “Has Tony asked about me?” she had asked in as casual a voice as she could manage.

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