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1.
RNA Biol ; 21(1): 1-20, 2024 01.
Article in English | MEDLINE | ID: mdl-38017665

ABSTRACT

The process of alternative splicing (AS) is widely deregulated in a variety of cancers. Splicing is dependent upon splicing factors. Recently, several long noncoding RNAs (lncRNAs) have been shown to regulate AS by directly/indirectly interacting with splicing factors. This review focuses on the regulation of AS by lncRNAs through their interaction with splicing factors. AS mis-regulation caused by either mutation in splicing factors or deregulated expression of splicing factors and lncRNAs has been shown to be involved in cancer development and progression, making aberrant splicing, splicing factors and lncRNA suitable targets for cancer therapy. This review also addresses some of the current approaches used to target AS, splicing factors and lncRNAs. Finally, we discuss research challenges, some of the unanswered questions in the field and provide recommendations to advance understanding of the nexus of lncRNAs, AS and splicing factors in cancer.


Subject(s)
Neoplasms , RNA, Long Noncoding , Humans , Alternative Splicing , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , RNA Splicing Factors/genetics , Neoplasms/genetics , RNA Splicing
2.
PLoS One ; 18(8): e0289813, 2023.
Article in English | MEDLINE | ID: mdl-37561696

ABSTRACT

The value of combining hybridization and mutagenesis in sesame was examined to determine if treating hybrid sesame plant material with mutagens generated greater genetic variability in four key productivity traits than either the separate hybridization or mutation of plant material. In a randomized block design with three replications, six F2M2 varieties, three F2varieties, and three parental varieties were assessed at Odisha University of Agriculture and Technology, Bhubaneswar, Odisha, India. The plant characteristics height, number of seed capsules per plant, and seed yield per plant had greater variability in the F2M2 generation than their respective controls (F2), however, the number of primary branches per plant varied less than in the control population. The chances for trait selection to be operative were high for all the characteristics examined except the number of primary branches per plant, as indicated by heritability estimates. Increases in the mean and variability of the characteristics examined indicted a greater incidence of beneficial mutations and the breakdown of undesirable linkages with increased recombination. At both phenotypic and genotypic levels strong positive correlations between both primary branch number and capsule number with seed yield suggest that these traits are important for indirect improvement in sesame seed yield. As a result of the association analysis, sesame seed yield and its component traits improved significantly, which may be attributed to the independent polygenic mutations and enlarged recombination of the polygenes controlling the examined characteristics. Compared to the corresponding control treatment or to one cycle of mutagenic treatment, two cycles of mutagenic treatment resulted in increased variability, higher transgressive segregates, PTS mean and average transgression for sesame seed yield. These findings highlight the value of implementing two EMS treatment cycles to generate improved sesame lines. Furthermore, the extra variability created through hybridization may have potential in subsequent breeding research and improved seed yield segregants may be further advanced to develop ever-superior sesame varieties.


Subject(s)
Sesamum , Sesamum/genetics , Plant Breeding , Phenotype , Genotype , Mutagenesis
3.
Agri ; 34(3): 210-212, 2022 Jul.
Article in English | MEDLINE | ID: mdl-35792699

ABSTRACT

Peripheral neuropathy secondary to entrapment of the nerves is not an uncommon etiology. Nerve entrapment is a common occurrence following trauma or surgery and poses significant diagnostic challenge. Entrapment neuropathy (EN) may not respond to standard neuropathic medication and may need invasive treatment. Pulsed radiofrequency (PRF) application is a recent modality and is gaining popularity for many EN as it does not cause neural ablation unlike conventional radiofrequency ablation. In this report, we present a case of young patient who presented with severe lower lateral leg pain in whom superficial peroneal nerve (SPN) EN was suspected and diagnostic SPN injection under ultrasound guidance confirmed the diagnosis. He subsequently underwent PRF neuromodulation and experienced long-lasting pain relief.


Subject(s)
Peroneal Neuropathies , Pulsed Radiofrequency Treatment , Humans , Male , Pain/etiology , Peroneal Neuropathies/diagnosis , Peroneal Neuropathies/etiology , Peroneal Neuropathies/therapy , Pulsed Radiofrequency Treatment/adverse effects , Ultrasonography
4.
Am J Hum Genet ; 109(8): 1549-1558, 2022 08 04.
Article in English | MEDLINE | ID: mdl-35858628

ABSTRACT

Deoxyhypusine hydroxylase (DOHH) is the enzyme catalyzing the second step in the post-translational synthesis of hypusine [Nε-(4-amino-2-hydroxybutyl)lysine] in the eukaryotic initiation factor 5A (eIF5A). Hypusine is formed exclusively in eIF5A by two sequential enzymatic steps catalyzed by deoxyhypusine synthase (DHPS) and deoxyhypusine hydroxylase (DOHH). Hypusinated eIF5A is essential for translation and cell proliferation in eukaryotes, and all three genes encoding eIF5A, DHPS, and DOHH are highly conserved throughout eukaryotes. Pathogenic variants affecting either DHPS or EIF5A have been previously associated with neurodevelopmental disorders. Using trio exome sequencing, we identified rare bi-allelic pathogenic missense and truncating DOHH variants segregating with disease in five affected individuals from four unrelated families. The DOHH variants are associated with a neurodevelopmental phenotype that is similar to phenotypes caused by DHPS or EIF5A variants and includes global developmental delay, intellectual disability, facial dysmorphism, and microcephaly. A two-dimensional gel analyses revealed the accumulation of deoxyhypusine-containing eIF5A [eIF5A(Dhp)] and a reduction in the hypusinated eIF5A in fibroblasts derived from affected individuals, providing biochemical evidence for deficiency of DOHH activity in cells carrying the bi-allelic DOHH variants. Our data suggest that rare bi-allelic variants in DOHH result in reduced enzyme activity, limit the hypusination of eIF5A, and thereby lead to a neurodevelopmental disorder.


Subject(s)
Lysine , Mixed Function Oxygenases , Neurodevelopmental Disorders , Alleles , Gene Expression , Humans , Lysine/analogs & derivatives , Mixed Function Oxygenases/genetics , Neurodevelopmental Disorders/genetics
5.
Ann Card Anaesth ; 25(1): 97-99, 2022.
Article in English | MEDLINE | ID: mdl-35075030

ABSTRACT

Persistent poststernotomy pain (PSP) is a well-known entity following cardiac surgery done with midline strenotomy. The severity of pain is usually mild to moderate in the majority of the patients. However, a small percentage of patients develop severe and persistent pain and need aggressive treatment. Our patient, a 63-year-old lady developed chronic severe parasternal pain following coronary artery bypass graft surgery. As multiple medications did not relieve her pain effectively, we did an ultrasound-guided pectoral-intercostal fascial plane block to which she responded with excellent and long-lasting pain relief. This is the first such case report of the use of this novel block technique for treating PSP.


Subject(s)
Cardiac Surgical Procedures , Chronic Pain , Nerve Block , Cardiac Surgical Procedures/adverse effects , Chronic Pain/therapy , Female , Humans , Middle Aged , Nerve Block/methods , Pain, Postoperative/drug therapy , Pain, Postoperative/etiology , Sternotomy/adverse effects
6.
Amino Acids ; 54(4): 485-499, 2022 Apr.
Article in English | MEDLINE | ID: mdl-34273022

ABSTRACT

Hypusine [Nε-(4-amino-2-hydroxybutyl)lysine] is a derivative of lysine that is formed post-translationally in the eukaryotic initiation factor 5A (eIF5A). Its occurrence at a single site in one cellular protein defines hypusine synthesis as one of the most specific post-translational modifications. Synthesis of hypusine involves two enzymatic steps: first, deoxyhypusine synthase (DHPS) cleaves the 4-aminobutyl moiety of spermidine and transfers it to the ε-amino group of a specific lysine residue of the eIF5A precursor protein to form an intermediate, deoxyhypusine [Nε-(4-aminobutyl)lysine]. This intermediate is subsequently hydroxylated by deoxyhypusine hydroxylase (DOHH) to form hypusine in eIF5A. eIF5A, DHPS, and DOHH are highly conserved in all eukaryotes, and both enzymes exhibit a strict specificity toward eIF5A substrates. eIF5A promotes translation elongation globally by alleviating ribosome stalling and it also facilitates translation termination. Hypusine is required for the activity of eIF5A, mammalian cell proliferation, and animal development. Homozygous knockout of any of the three genes, Eif5a, Dhps, or Dohh, leads to embryonic lethality in mice. eIF5A has been implicated in various human pathological conditions. A recent genetic study reveals that heterozygous germline EIF5A variants cause Faundes-Banka syndrome, a craniofacial-neurodevelopmental malformations in humans. Biallelic variants of DHPS were identified as the genetic basis underlying a rare inherited neurodevelopmental disorder. Furthermore, biallelic DOHH variants also appear to be associated with neurodevelopmental disorder. The clinical phenotypes of these patients include intellectual disability, developmental delay, seizures, microcephaly, growth impairment, and/or facial dysmorphisms. Taken together, these findings underscore the importance of eIF5A and the hypusine modification pathway in neurodevelopment in humans.


Subject(s)
Lysine , Oxidoreductases Acting on CH-NH Group Donors , Peptide Initiation Factors , RNA-Binding Proteins , Animals , Humans , Lysine/analogs & derivatives , Lysine/metabolism , Mice , Neurodevelopmental Disorders/genetics , Oxidoreductases Acting on CH-NH Group Donors/metabolism , Peptide Initiation Factors/metabolism , Protein Biosynthesis , Protein Processing, Post-Translational , Eukaryotic Translation Initiation Factor 5A
7.
J Biol Chem ; 297(5): 101333, 2021 11.
Article in English | MEDLINE | ID: mdl-34688659

ABSTRACT

Eukaryotic initiation factor 5A (eIF5A)†,‡ is an essential protein that requires a unique amino acid, hypusine, for its activity. Hypusine is formed exclusively in eIF5A post-translationally via two enzymes, deoxyhypusine synthase (DHPS) and deoxyhypusine hydroxylase. Each of the genes encoding these proteins, Eif5a, Dhps, and Dohh, is required for mouse embryonic development. Variants in EIF5A or DHPS were recently identified as the genetic basis underlying certain rare neurodevelopmental disorders in humans. To investigate the roles of eIF5A and DHPS in brain development, we generated four conditional KO mouse strains using the Emx1-Cre or Camk2a-Cre strains and examined the effects of temporal- and region-specific deletion of Eif5a or Dhps. The conditional deletion of Dhps or Eif5a by Emx1 promotor-driven Cre expression (E9.5, in the cortex and hippocampus) led to gross defects in forebrain development, reduced growth, and premature death. On the other hand, the conditional deletion of Dhps or Eif5a by Camk2a promoter-driven Cre expression (postnatal, mainly in the CA1 region of the hippocampus) did not lead to global developmental defects; rather, these KO animals exhibited severe impairment in spatial learning, contextual learning, and memory when subjected to the Morris water maze and a contextual learning test. In both models, the Dhps-KO mice displayed more severe impairment than their Eif5a-KO counterparts. The observed defects in the brain, global development, or cognitive functions most likely result from translation errors due to a deficiency in active, hypusinated eIF5A. Our study underscores the important roles of eIF5A and DHPS in neurodevelopment.


Subject(s)
Cerebellar Cortex/metabolism , Cognition , Hippocampus/metabolism , Mixed Function Oxygenases/metabolism , Neurogenesis , Neurons/metabolism , Oxidoreductases Acting on CH-NH Group Donors/metabolism , Peptide Initiation Factors/metabolism , RNA-Binding Proteins/metabolism , Animals , Humans , Lysine/analogs & derivatives , Lysine/metabolism , Mice , Mice, Knockout , Mixed Function Oxygenases/genetics , Organ Specificity , Oxidoreductases Acting on CH-NH Group Donors/genetics , Peptide Initiation Factors/genetics , RNA-Binding Proteins/genetics , Eukaryotic Translation Initiation Factor 5A
8.
Ann Card Anaesth ; 24(3): 415-418, 2021.
Article in English | MEDLINE | ID: mdl-34269285

ABSTRACT

Electrical storm or incessant ventricular tachycardia is a life-threatening condition and is associated with high morbidity and mortality. Often patients respond to traditional anti-arrhythmia treatment. However, some patients are resistant to the drug therapy and thus, pose huge challenges in effective management. Though stellate ganglion block has been found to be effective in treating patients with electrical storm, it is still under-utilized. In this case report, we successfully managed to revert the drug-resistant arrhythmia to sinus rhythm after ultrasound-guided stellate ganglion block. Earlier utilization of the block can possibly provide effective treatment in drug-resistant ventricular arrhythmias and prevent morbidity and mortality.


Subject(s)
Autonomic Nerve Block , Pharmaceutical Preparations , Tachycardia, Ventricular , Arrhythmias, Cardiac , Humans , Stellate Ganglion
10.
Nat Chem Biol ; 15(2): 123-131, 2019 02.
Article in English | MEDLINE | ID: mdl-30598543

ABSTRACT

Phosphorylation of the carboxyl-terminal domain (CTD) of the largest subunit of RNA polymerase II (Pol II) governs stage-specific interactions with different cellular machines. The CTD consists of Y1S2P3T4S5P6S7 heptad repeats and sequential phosphorylations of Ser7, Ser5 and Ser2 occur universally at Pol II-transcribed genes. Phosphorylation of Thr4, however, appears to selectively modulate transcription of specific classes of genes. Here, we identify ten new Thr4 kinases from different kinase structural groups. Irreversible chemical inhibition of the most active Thr4 kinase, Hrr25, reveals a novel role for this kinase in transcription termination of specific class of noncoding snoRNA genes. Genome-wide profiles of Hrr25 reveal a selective enrichment at 3' regions of noncoding genes that display termination defects. Importantly, phospho-Thr4 marks placed by Hrr25 are recognized by Rtt103, a key component of the termination machinery. Our results suggest that these uncommon CTD kinases place phospho-Thr4 marks to regulate expression of targeted genes.


Subject(s)
Protein Kinases/metabolism , RNA Polymerase II/genetics , RNA Polymerase II/physiology , Amino Acid Sequence , Casein Kinase I/metabolism , Phosphorylation , Phylogeny , Protein Domains , Protein Serine-Threonine Kinases/metabolism , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/metabolism , Threonine/metabolism , Transcription, Genetic
11.
Indian J Ophthalmol ; 65(12): 1431-1435, 2017 Dec.
Article in English | MEDLINE | ID: mdl-29208829

ABSTRACT

PURPOSE: To compare Endothelial cell(EC) loss following Phacoemulsification (PKE) in pupils of different sizes. METHODS: A prospective double masked observational study in which a total of 150 eyes of 150 patients between 50 & 70 years of age with senile cataract of nuclear sclerosis grade II were enrolled. Patients were allocated into three groups of 50 eyes each in Group A (pupil size <5 mm), Group B (pupil size 5-7 mm) and Group C (pupil size >7 mm). Pupillary size was measured by determining the height of slit on slit-lamp biomicroscope examination. PKE was done by the same expert surgeon using vertical chop technique and a foldable intraocular lens was implanted in the capsular bag. Corneal EC count and pachymetry were performed twice and average of 2 readings was taken for the purpose of this study. Measurements were taken preoperatively and postoperatively on day 1, day 7 and day 30. RESULTS: The mean EC count loss on postoperative day 1 in Group A was 19.45%, Group B 14.89%, Group C 10.19% with statistical significant difference between Group A and Group B, as also Group A and Group C. The difference was not significant between Group B and Group C, though there was a fall in EC count in Group C as well. Increase in corneal thickness on postoperative day 1 in group A was 5.43%, Group B 3.55%, Group C 2.14% with statistical significant difference between Group A and Group B, as also Group A and Group C with no difference in Group B and Group C. CONCLUSION: PKE done in eyes with maximal pupillary dilatation of <5 mm causes a greater EC loss and results in thicker corneas postoperatively as compared to eyes with pupillary dilatation of >5 mm at the end of one month.


Subject(s)
Corneal Diseases/diagnosis , Endothelium, Corneal/pathology , Phacoemulsification/adverse effects , Postoperative Complications/diagnosis , Pupil , Aged , Cell Count , Corneal Diseases/etiology , Double-Blind Method , Female , Follow-Up Studies , Humans , Male , Middle Aged , Postoperative Complications/etiology , Prospective Studies , Slit Lamp Microscopy/methods , Time Factors , Visual Acuity
12.
J Mol Biol ; 429(1): 158-176, 2017 01 06.
Article in English | MEDLINE | ID: mdl-27913116

ABSTRACT

Gal3p is an allosteric monomeric protein that activates the GAL genetic switch of Saccharomyces cerevisiae in response to galactose. Expression of constitutive mutant of Gal3p or overexpression of wild-type Gal3p activates the GAL switch in the absence of galactose. These data suggest that Gal3p exists as an ensemble of active and inactive conformations. Structural data have indicated that Gal3p exists in open (inactive) and closed (active) conformations. However, a mutant of Gal3p that predominantly exists in inactive conformation and is yet capable of responding to galactose has not been isolated. To understand the mechanism of allosteric transition, we have isolated a triple mutant of Gal3p with V273I, T404A, and N450D substitutions, which, upon overexpression, fails to activate the GAL switch on its own but activates the switch in response to galactose. Overexpression of Gal3p mutants with single or double mutations in any of the three combinations failed to exhibit the behavior of the triple mutant. Molecular dynamics analysis of the wild-type and the triple mutant along with two previously reported constitutive mutants suggests that the wild-type Gal3p may also exist in super-open conformation. Furthermore, our results suggest that the dynamics of residue F237 situated in the hydrophobic pocket located in the hinge region drives the transition between different conformations. Based on this study, we suggest that conformational selection mechanism is the driving force in the allosteric transition of Gal3p, which may have implications in other signaling pathways involving monomeric proteins.


Subject(s)
Galactose/metabolism , Saccharomyces cerevisiae Proteins/chemistry , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae/enzymology , Saccharomyces cerevisiae/metabolism , Transcription Factors/chemistry , Transcription Factors/metabolism , Allosteric Regulation , Amino Acid Substitution , Models, Molecular , Molecular Dynamics Simulation , Mutant Proteins/chemistry , Mutant Proteins/genetics , Mutant Proteins/metabolism , Protein Conformation , Saccharomyces cerevisiae/growth & development , Saccharomyces cerevisiae Proteins/genetics , Transcription Factors/genetics
13.
J Clin Diagn Res ; 10(11): MC04-MC06, 2016 Nov.
Article in English | MEDLINE | ID: mdl-28050413

ABSTRACT

INTRODUCTION: "As the septum goes, so goes the nose". A well-known phrase by Maurice Cottle forms the pillar of septoplasty. Since the inception of septal surgeries, numerous methods of septoplasty have been described. But, if not performed meticulously, may lead to deformity. For a successful surgery, understanding the anatomy and addressing the anterior nasal spine and maintaining the tip integrity is vital. AIM: To study the outcomes of "ROUND ABOUT technique" to correct deviated nasal septum which focuses on the importance of anterior spine and hence maintain the tip integrity. MATERIALS AND METHODS: This was a prospective, single-centre outcome study of 35 patients with symptomatic nasal obstruction. Here, we describe a method of elevating the mucoperichondrial and mucoperiosteal flaps bilaterally, without transecting the quadrilateral cartilage of the septum. The Sino Nasal Outcome Test-22 (SNOT-22) Questionnaire was administered pre-operatively and after 3 months following surgery. The post-operative follow-up period ranged from 3 to 6 months (mean= 4.5 months) to evaluate the functional and aesthetic outcomes of the performed procedure. RESULTS: A total of 35 patients underwent surgery by this technique who presented with deviated nasal septum and variable degrees of nasal obstruction. To assess the statistical outcome, Paired t-test was applied. Mean SNOT-22 scores decreased significantly from 40.02 pre-operatively to 18.65 three months after surgery. The results sustained after 6 months (p-value <0.0001), 85% of these patients had improved breathing post-operatively and none of the patients complained any aesthetic criticisms. The patients were content and the requirement of medications post-operatively were minimal. CONCLUSION: The ROUND ABOUT technique is a very effective and safe method in correcting the septal deviations especially the ones with caudal or dorsal deflections. It also helps in maintaining the tip integrity and addressing the anterior nasal spine. It avoids the complications of conventional septoplasty.

14.
J Clin Diagn Res ; 9(9): XD01-XD03, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26500997

ABSTRACT

Langerhans cell histiocytosis (LCH) is a rare disorder of mononuclear phagocytic system whose clinical presentation varies from the localised involvement of a single bone to a widely disseminated disease. Langerhans cell histiocytosis rarely involves the thyroid gland and isolated involvement of thyroid is even rarer. We report a case of an eight-year-old male child diagnosed with Langerhans cell histiocytosis limited to thyroid gland with review of literature. Should thyroidectomy be done or proceed with chemotherapy? Our case report raises this question with a note on the role of 18 fluoro deoxy glucose positron emission tomography-computed tomography in the management of the same.

15.
Mol Microbiol ; 94(1): 202-17, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25135592

ABSTRACT

In S. cerevisiae, following the Whole Genome Duplication (WGD), GAL1-encoded galactokinase retained its signal transduction function but lost basal expression. On the other hand, its paralogue GAL3, lost kinase activity but retained its signalling function and basal expression, thus making it indispensable for the rapid induction of the S. cerevisiae GAL switch. However, a gal3Δ strain exhibits delayed growth kinetics due to the redundant signalling function of GAL1. The subfunctionalization between the paralogues GAL1 and GAL3 is due to expression divergence and is proposed to be due to the alteration in the Upstream Activating Sequences (UASG ). We demonstrate that the GAL switch becomes independent of GAL3 by altering the interaction between Gal4p and Gal80p without altering the configuration of UASG . In addition to the above, the altered switch of S. cerevisiae loses ultrasensitivity and stringent glucose repression. These changes caused an increase in fitness in the disaccharide melibiose at the expense of a decrease in fitness in galactose. The above altered features of the ScGAL switch are similar to the features of the GAL switch of K. lactis that diverged from S. cerevisiae before the WGD.


Subject(s)
DNA-Binding Proteins/metabolism , Galactose/metabolism , Glucose/metabolism , Repressor Proteins/metabolism , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae/metabolism , Transcription Factors/metabolism , DNA-Binding Proteins/genetics , Gene Expression Regulation, Fungal , Protein Binding , Repressor Proteins/genetics , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae Proteins/genetics , Transcription Factors/genetics
16.
FEBS J ; 281(7): 1798-817, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24785355

ABSTRACT

GAL1 and GAL3 are paralogous signal transducers that functionally inactivate Gal80p to activate the Gal4p-dependent transcriptional activation of GAL genes in Saccharomyces cerevisiae in response to galactose. Unlike a wild-type strain, the gal3∆ strain shows delayed growth kinetics as a result of the signaling function of GAL1. The mechanism ensuring that GAL1 is eventually expressed to turn on the GAL switch in the gal3∆ strain remains a paradox. Using galactose and histidine growth complementation assays, we demonstrate that 0.3% of the gal3∆ cell population responds to galactose. This is corroborated by flow cytometry and microscopic analysis. The galactose responders and nonresponders isolated from the galactose-adapted population attain the original bimodal state and this phenotype is found to be as hard wired as a genetic trait. Computational analysis suggests that the log-normal distribution in GAL4 synthesis can lead to bimodal expression of GAL80, resulting in the bimodal expression of GAL genes. Heterozygosity at the GAL80 but not at the GAL1, GAL2 or GAL4 locus alters the extent of bimodality of the gal3∆ cell population. We suggest that the asymmetric expression pattern between GAL1 and GAL3 results in the ability of S. cerevisiae to activate the GAL pathway by conferring nongenetic heterogeneity.


Subject(s)
Mutation , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae/metabolism , Transcription Factors/metabolism , Transcriptional Activation , Galactokinase/genetics , Galactokinase/metabolism , Galactose/metabolism , Heterozygote , Monosaccharide Transport Proteins/genetics , Monosaccharide Transport Proteins/metabolism , Repressor Proteins/genetics , Repressor Proteins/metabolism , Saccharomyces cerevisiae/enzymology , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae Proteins/genetics , Stochastic Processes , Transcription Factors/genetics
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