This time its the big man’s story

Vidrum was hurrying back to his office after a quick dinner. As he did so he mentally rued the fact that every time he found his way out of one problem in life a new one came upon him. Once in his office he rummaged some of the drawers to find the documents he was looking for; but they were not there. He was puzzled, he clearly remembered putting them back into the drawers after a conversation with a visitor earlier in the day. Holding his head in despair, he recalled that after meeting his visitor in the morning he had gone on deliver consultation to his patients. Hence, he decided to head down two floors to the rooms where he saw his patients. As walked down the corridor he saw the light in Vrishchika’s office to be on and heard her talking to someone with a familiar voice [Footnote 1]. It was not unusual to see Vrishchika at that time because since having her child she saw patients only till noon and came back in the evenings to do her research. But Vidrum was rarely around in the evenings and if he did stay on or come in it was because he had to attend to something that needed attention. To his surprise he found that Vrishchika was with Lootika in her office and they were looking at some images being streamed in from her microscope.

Vidrum: “Don’t want to disturb you ladies, but hope you two are doing well. Lootika it is a real surprise to see you here in the medical school campus. What brings you here?”
Lootika: “Good to see you after long; we are surviving and hope life ain’t treating you too badly.”
Vidrum then added with a grin: “Lootika I thought you would be working on more interesting organisms than Homo sapiens.”
Lootika: “Now our husbands our working together on more interesting organisms and Varoli, who has just started her lab, has also joined forces with them. I have hence turned to the conquest of uncharted territory in the biochemistry physiology of Homo along with sis and the combined forces of our pupils.”
Vrishchika: “Like emperor Raghu’s digvijaya we have embarked on a campaign of the human cell, all the way from the nucleus to the extracellular matrix to bring a closure to the proteins C6orf70, leprecan, FAM55D PC-esterase, C5ORF32, and C7orf58.” Pointing to her screen she continued: “See these are promyelocytes in which we have induced the expression of C5ORF32 and they are surviving multiple chemical stressors better that those in which it has not been induced.”
Vidrum: “As ever it looks like you have something interesting to keep yourselves busy with. But I have my more mundane troubles to return to.”

Lootika and Vrishchika: “Why, what is troubling you?”
Vidrum: “Well key documents concerning the experimental treatment I was managing are missing from my office. I remember so clearly putting them in my drawer. That’s why I am on my way to check if I left them in the consulting rooms. On top of that I just heard this evening that our senior colleague Prof. Suryaprakash is seriously ill with the unexpected onset of multiple neurological deficits. Just before he took ill he was supposed to tell me something important about my own trial for he was doing something comparable, although with other tumors.”

Vrishchika: “That is awful – do you have any details? It is tragic to hear that so strong and intellectually agile a man has been suddenly disabled thus. It reminds us of our own impending mortality. I have indeed not seen Suryaprakash for several days. Now I recall that he did look a little unsure while taking the stairs when I last saw him. But then I did not give it much thought. Thus indeed are the ways by which the black son of Vivasvān drags his victims with his pāśa. By the way I do hope you find your documents soon.”
Vidrum: “I am going to see him tomorrow morning…will let you know” Saying so Vidrum scurried along to search for his documents.”

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Later that week Indrasena was surprised to see his wife Vrishchika burst in to his lab as she seldom visited him at work. She frantically asked: “Ārya, is Lootika around?”
Indrasena: “Did you check in her lab (Lootika’s lab was next to that of Indrasena)? Gautamī, is something amiss you look bothered?”
Vrishchika: “She is not there. I need to see agrajā right away I will explain things to you later at home.”
Indrasena: “She must have gone home to attend to the kids but if it is some real problem you could go to Somakhya’s office and tell him.”
Vrishchika: “Don’t worry it is nothing for you to be concerned about now. This is something that only she would understand at this point. For now I will just say that it about ZU5 and DEATH-like domains.”
Vrishchika next hastened to Somakhya’s office and without any introduction said in voice barely concealing her agitation: “Where is agrajā?”
Somakhya: “Why, is there some problem? She must have gone home if you are not finding her here. What has brought you so unexpectedly to our campus?”
Vrishchika: “Lootika will tell you everything, but let me go and see her. It is not something I should bother you with while at work.” Saying so she hurried away.

Surprised by this appearance of Vrishchika and her cryptic statements, Somakhya went over to Indrasena and said: “Vrishchika was here asking for Lootika. I sensed some discomposure in her voice and appearance. Any trouble?”
Indrasena: “Yes, I too was puzzled. All she told me was it is about ZU5 and DEATH-like domains. Wonder why that should produce such an effect on her. I know she is not currently working on any genes coding for with these.”
Somakhya: “Currently, nor is Lootika; though there was stuff she did with the proteins with those domains from the boring sponge Zyzzya which we two had pulled off from the Lakṣadvīpa-s. She mentioned reading our paper on that and being inspired by it in the period when were out of contact.”
Indrasena: “Strange indeed. I don’t quite understand why this should be a matter of concern for our patnī-s. In any case I guess we would learn of it soon, but I am sure if it was really something of a bother, needing our attention, they would have told us of it.”

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In the meantime Vrishchika picked up her kid and went to see Lootika at home. As she settled down on the chair she said: “agrajā, I heard the details of the tragic case of Prof. Suryaprakash from Vidrum. He seems to have contracted a disease with several manifestations suggestive of Creutzfeldt–Jakob disease. However, they failed to detect the PrPSc despite finding 14-3-3 proteins in his cerebrospinal fluid. But everything else is symptomatically comparable. As far as we know, we do not have any evidence he consumed meat or had an invasive procedure or transfusion that could have transmitted it. Lootika, what does this bring to your mind?”

Lootika: “That is tragic indeed. He had been good to you and was a good medical scientist. But you are the physician here; anujā, so why do you ask me – there must be something more to this if you have come running all way from to see me, since we would have any how met in the evening. Perhaps it was one of those other diseases coming from Prnp mutations deleting parts of the N-terminal region or it could have been an octapeptide expansions and may be they failed to detect it?”

Vrishchika: “Yes all that came to mind when Vidrum informed me but they have all been ruled out. In fact his Prnp is totally normal!”
Lootika: “Then it is something else which is resulting in similar symptoms. May be there is some issue with the doppel protein or the shadoo protein?”
Vrishchika: “They did quite a thorough job and checked for those – there was no mutation or polymorphism in those genes and we have no evidence that their expression changes can induce prions.”

Lootika: “There are other neurodegenerative conditions – some type of aggregation disorder I would postulate.”
Vrishchika: “Yes possible. But upon hearing the case from Vidrum, I did intervene right away and carried out a panel of tests for repeat expansions and possible protein aggregation disorders. All negative. But one thing struck in the biopsy material from the enteric nervous system. Rampant neuron death due to polymers of DLD proteins, primarily those of the NACHT ATPase NALP1. Incidentally, I used Varoli’s new direct AAA+ motor-peptidase coupled protein sequencing method – it worked brilliantly. Now an intracellular prion like transformation cannot easily pathologically transmit like a glycophosphatidylinositol anchored PrPSc, which encounters abundant PRNP protein at the cell surface. So what do you think is happening here Lootika?”
Lootika: “Well that is rather surprising. It reminds me of what my engineered Sarcocystis neurona could do to its victim. That is exactly why we had used it to probe the nervous system.”
Vrishchika: “That is why I am here, agrajā. Your engineered S.neurona is the one that can do it – that with the macrophage chitinase inhibitor which injects the sponge ZU5-DEATH protein, one of those you learned of from our puruṣa-s. I cannot forget your words when you successfully made it – ‘this could be a biological weapon that might be useful when that great irruption of dasyu-s threatens our very existence.’ But who could have used it like this against Suryaprakash?”

Lootika: “Vrishchika, my strains are under lock-and-key with access only by biometric scan and is under video surveillance. Moreover, all my weaponizable constructs are kept separately and labeled using a cipher the key for which is only known to me and your husband. Other than he and I, only one technician maintaining the cells and parasites has access to the room. The technician is reliable and would not know what is what unless she chose to take out all the apicomplexans in the collection. If she were to do that she would be caught by the scanner. Are you sure it could be one of my bugs?”

Vrishchika: “Wow, I forgot all the weaponizable material was well-guarded. But who else could have it other than you? What about…”
Lootika: “When I had done those experiments I had an upacīna student who did lot of the initial animal work; Other than me she can make those strains in principle. She went on to become a post-doc with Sterling McKnight that dhūrta who deceivingly set up a collaboration with me and then tried to run away with our discoveries and publish them as his. You may recall how the two of us then narrowly outran the mleccha rogue in that race. That dhūrta McKnight does know how to make those organisms and I have reason to believe he has made several of them for the mleccha-senā. But then he is two oceans away from us and what reason does he have to target Suryaprakash!”

Vrishchika: “I hear all your objections. I will check more carefully again. That is why I need the S.neurona primers from you. We were unable to microscopically detect the organism.”
Lootika: “I can give you the S.neurona primers later today. But we cannot rule out incidental Sarcocystis infection. As you know better than me people do get it once in a way.”
Vrishchika: “But is there some conclusive test?”
Lootika: “anujā, think carefully – Somakhya’s mother thinks you are the the smartest of the four of us – do I need to be telling you of all people about monocytes [Footnote 2]”
Vrishchika: “Ah – I believe I should be able to get the bug out by establishing a monocyte culture and sorting for CD11b positive cells. Then we will sequence the genome and see if and how it is has been engineered. ”
Lootika: “Yes. That would be the ultimate test. A skilled molecular biologist could use the details we provided in our paper to re-engineer such a such a bug with different specifications. We know if that was really done.”
Vrishchika: But let us make sure this evening that there has been no tampering of your biological storage. I just needed to tell you this for I think there is more to it that we know right now. Now let me go back home – I need to get something ready for dinner.”

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Vrishchika received a mail from the chairman of the medical school:
“We are interviewing Dr. Brihat Gupta for a faculty position (CV is attached). The search committee would particularly value your opinion on him especially given that he combines a strong basic research component with his clinical work. He comes from the lab of the renowned cancer biologist Prof. Richard Junkberg and as you can see has an impressive list of highly cited publications. All in all, he is seen as a rising star in experimental therapeutics and we would really appreciate if you attend his talk. The time and abstract of his talk are appended below.”

Vrishchika glanced through his abstract and seeing the phrase “…therapeutic maneuvering of social RNA in the war against cancer…” thought to herself: “This must be given close attention wonder what he has to say.”

On the day of Brihat Kumar Gupta’s talk Vrischika arrived at the auditorium and noticed that there was a swelling crowd. There were students who were already trying to sneak an introduction to the speaker, even as he was girt by the search committee and brought to the auditorium. He was introduced to Vrishchika as he was being ushered in by her colleagues. In an unadulterated mleccha accent he professed knowledge of her work and smoothly complimented her for some publication in a high profile tabloid. Vrishchika being her usual shy self in public shrugged it away and merely said she looked forward to his talk.

While Gupta’s clinical exploits looked impressive, Vrishchika found his basic research component to be heavy on phenomenology, with little to offer in terms of crisp demonstration of precise biochemistry, which her sister and Somakhya had instilled in her from childhood. As her eyes were almost shutting down in the dim light of the auditorium, she suddenly perked up at the display of a miRNA supposed to inhibit the HoxD10 gene. The alignment looked so flimsy that it was rather unlikely that HoxD10 was ever a significant target for that miRNA. She surreptitiously took a photo of the slide and sent it over to Indrasena asking if it made sense. This made her pay closer attention to what Gupta was presenting, when suddenly she saw a data figure on his slide where two bands seemed exactly the same. But they had been pasted at different positions as though they were reporting the expression of two different genes on the gel. She remarked to herself: “That looks very strange indeed!” As Gupta went on he said: “This miRNA is showing great promise and we developed therapeutic RNAs in collaboration with Prof. Sterling McKnight, which were being subject to a large trial by Dr. Suryaprakash in your department. I am grieved to hear that he has recently taken seriously ill. However, on the positive side a parallel trial by the team put together by Prof. McKnight at his medical school have yielded excellent results and were just published. Vidrum in your school is also managing the trials of another class of therapeutic RNAs designed in Prof. Junkberg’s lab. I hear Dr. Sonenberg from his lab had just visited your school recently in this regard. ”

As the talk concluded Vrishchika was tempted to ask Gupta about those figures but it suddenly hit her then that she was staring a something which might be linked to the other matter which had startled her some days back. Hence, she kept quiet. A throng of students surrounded Dr. BK Gupta, each vying for his attention, and some seeing him as an outside chance to make it to the lab of the great Prof. Junkberg. Just then Vrishchika’s student came up to her and asked her if it might be worth exploring such pleotropic miRNAs. Vrishchika: “Look more closely at his work and the data there might be something more than what it seems.” The chairman prised out Dr. Gupta from the cordon of students that had formed around him saying that he needed to go to his next meeting with faculty. As they were walking him to his next appointment, the chairman separated from the rest of the crowd a bit and turning to Vrishchika said: “Did you like the talk”
Vrishchika: “The talk had several notable points and we can talk about it at length when time permits.”

Realizing that she might not want to talk about it with Dr. Gupta within earshot they rejoined the rest of the group and the chairman asked: “Vrishchika, would you like to join us for dinner with Dr. Gupta.”
Vrishchika: “That would have been nice but I need to be at home then.” Vidrum just then chimed in: “Vrishchika, the restaurant we are going to should suit your dietary restrictions.” Hearing this Dr. Gupta, who was beside Vidrum, in his crisp accent added: “With South Asian food we can all ways find something for every palate.” Just then another colleague Ahmed al Zaman said with a smirk, his voice sounding like metal grating on metal: “You see she is a high caste brahmin; they don’t wine and dine with us groundlings.”

One part of Vrishchika wanted her to lash out in response to all this, but realizing that sometimes it was good thing to keep the sting hidden, she stated with a coy smile: “I have a little kid who needs some attention, else I would have been there with you all.” Most of her colleagues murmured sympathetically and proceeded their way. Vrishchika felt an urge to talk right away with her gang about the tangled web she had stumbled upon. But as she turned it over she realized there were many incomplete points and it was better she investigated the various leads a little more before letting the rest know. She also realized that it was not right to bother them at work with a matter which was still largely in the realm of gossip and decided for the time being she would only tell Lootika later in the evening. As she was returning home she saw a mail from Indrasena saying that indeed HoxD10 was unlikely to be a target for that supposed miRNA.

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That evening as Vrishchika and Lootika were riding to work the former filled in her sister of the Suryaprakash case. Everything was suggesting that her hunch, outlandish as it seemed to Lootika, was right. She had managed to conclusively show S.neurona in the patient’s monocyte culture and had sent its genome for sequencing. The sequence was to be ready for them to examine that evening.

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Footnote 1: Vrishchika was then a professor in the section for experimental therapeutics where Vidrum was also a consulting physician.

Footnote 2: That is the catur-bhaginī

continued…