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Tyagi, P.

Publications and source records attributed to Tyagi, P..

13 recordsLinked to original sources

Toxicity by Consortium Revisited: Structural Neighbors and Cross-Family Reach

A compound's toxicity can be gauged from the toxicities of the compounds it resembles. We showed that in 2003 for rat oral LD50: the compounds in a reference set of 13,645 that most resemble the query form a consortium and vote on its toxicity. This work returns to that method twenty-three years later, on public data, because the reference set used in 2003 is no longer public. Rebuilt on 11,989 curated rat oral LD50 values, the method answers for 67.0% of the held-out compounds carrying an exact value, and 92.1% of those answers land within one log unit; in 2003 the figures were 94% within one log unit on 57.3% of compounds. We then ran it over 28 further endpoints, each recording whether a compound showed a particular toxicity, from cytochrome inhibition to mutagenicity to clinical toxicity: 149,224 measurements on 34,162 compounds. The method answers only when a compound has close enough neighbors, and on those it correctly identifies a median 72.9% of the compounds that showed the toxicity and 96.5% of those that did not. Separately we asked whether a reverse screen carries information about these endpoints beyond molecular size and lipophilicity. We search a compound against a published index of 27,910 ligands solved inside proteins and count the protein families those sites belong to. A compound whose matches reach no protein family inhibits CYP3A4 2.5% of the time; one whose matches span sixteen or more families inhibits 51.0% of the time. Added to heavy atom count, molecular weight and cLogP, the screen raises the mean area under the ROC curve from 0.72 to 0.76 across the 28 endpoints. The OpenADMET consortium has since measured CYP3A4 inhibition on 2,335 compounds, and the same pattern holds. Compounds that resemble known binders of sixteen or more protein families inhibit CYP3A4 24.60% of the time; compounds that resemble no known binder at all inhibit it 5.00% of the time. The more protein families a compound resembles binders of, the more likely it is to inhibit CYP3A4. This association is strong enough to act on before anything is made. Scoring a library on how far its compounds reach across protein families, together with their size and lipophilicity, and setting aside the riskiest fifth removes two thirds of the androgen receptor binders and about half of the CYP3A4 inhibitors. Scoring needs only a two-dimensional structure, with no docking and no conformer generation at query time. Interactive scoring, the data and the pool are at toxpred.ai. Keywords: toxicity prediction; structural similarity; nearest neighbors; reverse screening; target breadth; promiscuity; LD50; cytochrome P450; virtual library triage

bioinformatics↗

Taguchi Design of Experiment for Optimizing Saline Solution Composition to Increase Electrical Coupling in a Multi-Electrode External EEG Sensor

The effectiveness of noninvasive brain sensing lies in the development of EEG (electroencephalogram) sensors capable of delivering high signal-to-noise ratios for real-time brain-machine interfacing and neuromodulation. A critical factor for achieving this goal is the optimization of electrical coupling between the skin and external electrodes, especially when detecting weak brain signals in the millivolt to microvolt range. In this study, we employed a Taguchi Design of Experiment (DoE) methodology to optimize the composition of an electrolyte solution used to enhance signal quality in a 14-electrode wearable EEG sensor system. We investigated four electrolyte components Sodium Chloride (NaCl), Potassium Chloride (KCl), Sodium Bicarbonate (NaHCO3), and Tween-20 surfactant each varied across three concentration levels using an L9 orthogonal array. For each experimental condition, ten measurements were collected to ensure statistical reliability. Two statistical analytical approaches were applied: average signal strength analysis, which exhibited considerable variability and limited diagnostic value, and signal-to-noise ratio (SNR) analysis, which used logarithmic scaling to provide more meaningful insights by minimizing noise effects. Results underscore that while the three salts made comparable contributions, the Tween-20 surfactant was the dominant factor affecting EEG signal strength. Additionally, all four factors exhibited non-linear behavior across concentration levels. This work presents a systematic optimization approach for improving electrolyte formulations and offers practical guidance for enhancing electrical coupling in EEG systems. The results support the futuristic development of high-SNR nanoscale EEG technologies where skin-sensor coupling will be highly sensitive towards saline solution as a conductive medium.

scientific communication and education↗

Bacterial extracellular vesicles as a tunable platform for vaginal drug delivery

There is a critical gap in the development of new therapeutic platforms designed to treat gynecologic and obstetric diseases. Compared to systemic drug delivery, vaginal drug administration of nanoparticle formulations limits off-target side effects while increasing therapeutic concentration in target tissues, showing promise for clinical translation. However, these formulations suffer from limited scalability, high-cost reagents, and long optimization timelines. Recent work highlights the potential of bacterial extracellular vesicles (bEVs) as a low-cost, tunable platform for therapeutic applications. Here, we evaluate bEVs as a therapeutic carrier for vaginal drug delivery. We demonstrate the loading of the model protein moxNeonGreen into Escherichia coli Nissle 1917 bEVs. By optimizing growth parameters, we increase protein loading into bEVs. We evaluate the effect of bEVs on the vaginal microenvironment, and observe no negative impact on vaginal epithelial cells, endocervical cells, or vaginal bacteria in vitro. Additionally, we observe the retention of bEVs in the murine female reproductive tract for more than six hours. This study provides a framework for using genetically engineered bEVs to rapidly generate customizable therapies for a range of gynecologic and obstetric conditions, addressing longstanding challenges in womens health therapeutics. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=50 SRC="FIGDIR/small/658669v1_ufig1.gif" ALT="Figure 1"> View larger version (19K): org.highwire.dtl.DTLVardef@e483d2org.highwire.dtl.DTLVardef@8a6d2corg.highwire.dtl.DTLVardef@5756borg.highwire.dtl.DTLVardef@5fdb67_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

DNA methylation and demethylation driven regulation of sepsis

During dysregulated inflammation and sepsis, there is a sudden surge of cytokines, such as TNF alpha, IL6, and IL10, which disrupts the body homeostasis. This sudden and rapid increase in cytokine gene expression cannot be explained by the conventional central dogma mechanism. DNA methylation is one of the most significant epigenetic modifications. DNA methylation is mostly associated with the suppression of gene activity, and DNA demethylation is associated with the activation of gene activity. There are numerous transcription factors, such as CREB1, c-FOS, AP-1, IRF1, and EGR1, that play pivotal roles in regulating inflammation. Thus, it was hypothesized that changes in the DNA methylation patterns of cytokine promoter regions or in the promoter regions of these specific transcription factors might be the potential reasons for the increase in cytokine levels during sepsis. During endotoxin stimulation, DNA methyltransferases were suppressed, and DNA methylation levels were altered at the global level. Bisulfite sequencing revealed no change in the DNA methylation patterns of the TNF-alpha, IL6 and IL10 cytokine promoters. The CREB1 and c-FOS transcription factor promoters were demethylated after LPS stimulation and in clinical sepsis samples. Overall, this study highlights the importance of the role of DNA methylation in sepsis.

molecular biology↗

Cocaine-induced DNA-PK relieves RNAP II pausing by promoting TRIM28 phosphorylation.

Drug abuse continues to pose a significant challenge in HIV control efforts. In our investigation, we discovered that cocaine not only upregulates the expression of DNA-dependent protein kinase (DNA-PK) but also augments DNA-PK activation by enhancing its phosphorylation at S2056. Moreover, DNA-PK phosphorylation triggers the translocation of DNA-PK into the nucleus. The finding that cocaine promotes nuclear translocation of DNA-PK further validates our observation of enhanced DNA-PK recruitment at the HIV long terminal repeat (LTR) following cocaine exposure. By activating and facilitating the nuclear translocation of DNA-PK, cocaine effectively orchestrates multiple stages of HIV transcription, thereby promoting HIV replication. Additionally, our study indicates that cocaine-induced DNA-PK promotes hyper-phosphorylation of RNA polymerase II (RNAP II) carboxyl-terminal domain (CTD) at Ser5 and Ser2 sites, enhancing both initiation and elongation phases, respectively, of HIV transcription. Cocaines enhancement of transcription initiation and elongation is further supported by its activation of cyclin-dependent kinase 7 (CDK7) and subsequent phosphorylation of CDK9, thereby promoting positive transcriptional elongation factor b (P-TEFb) activity. We demonstrate for the first time that cocaine, through DNA-PK activation, promotes the specific phosphorylation of TRIM28 at Serine 824 (p-TRIM28, S824). This modification converts TRIM28 from a transcriptional inhibitor to a transactivator for HIV transcription. Additionally, we observe that phosphorylation of TRIM28 (p-TRIM28, S824) promotes the transition from the pausing phase to the elongation phase of HIV transcription, thereby facilitating the production of full-length HIV genomic transcripts. This finding corroborates the observed enhanced RNAP II CTD phosphorylation at Ser2, a marker of transcriptional elongation, following cocaine exposure. Accordingly, upon cocaine treatment, we observed elevated recruitment of p-TRIM28-(S824) at the HIV LTR. Overall, our results have unraveled the intricate molecular mechanisms underlying cocaine-induced HIV transcription and gene expression. These findings hold promise for the development of highly targeted therapeutics aimed at mitigating the detrimental effects of cocaine in individuals living with HIV. Highlights of the studyO_LICocaine upregulates both the expression and activity of DNA-PK. C_LIO_LICocaine augments the phosphorylation of DNA-PK selectively at S2056, a post-translational modification that marks functionally active form of DNA-PK. C_LIO_LICocaine enhances the nuclear translocation of DNA-PK. C_LIO_LIThe DNA-PK inhibition severely impairs HIV transcription, replication, and latency reactivation. C_LIO_LICocaine facilitates the initiation and elongation phases of HIV by enhancing RNAPII CTD phosphorylation at Ser5 and Ser2, respectively, by stimulating DNA-PK. C_LIO_LICocaine also supports initiation and elongation phases of HIV transcription by stimulating CDK7 (the kinase of TFIIH) and CDK9 (the kinase subunit of P-TEFb), respectively. C_LIO_LICocaine-mediated activation of DNA-PK relieves RNAP II pausing by reversing the inhibitory effect of pausing factor TRIM28 and converting it into a transactivator by catalyzing its phosphorylation at S824 site. C_LIO_LIThus, cocaine, by activating DNA-PK, facilitates the multiple phases of HIV transcription, namely, initiation, RNAP II pause-release, and elongation. C_LI

molecular biology↗

De novo Sequencing and Whole Transcriptome Analysis for Biosynthesis Pathway of Diosgenin in Pedalium murex L.: A Medicinal herb

Transcriptome-based investigations of candidate genes, critical pathways, and gene regulation in non- model species have been transformed by high-throughput RNA sequencing in different studies. Pedalium murex L. is one of the most important annual medicinal herbs of the Pedaliaceae family. Because of its anti-inflammatory, antioxidant, antimicrobial, and anticancerous properties, P. murex is widely used in traditional medicine to treat gonorrhoea, leucorrhoea, urinary disorders, gastrointestinal tract disorders, cough, and asthma. Steroidal diosgenin is the major bioactive compound of P. murex. However, transcriptional technologies have yet to be used to study the steroid diosgenin biosynthetic pathway of this herb. In this study, we performed a whole transcriptomic analysis of the root, fruit, and leaf tissues of P. murex with three biological replicates and obtained [~]6.77 Gb of clean raw data. A total of 148871 unigenes were assembled with an average length of N50 and 1167 bp. Putative functions could be annotated to 75198 unigenes based on a BLASTX search against the NR, Uniprot, KEGG, Pfam, GO, and COG databases. Most of the unigenes related to steroidal diosgenin backbone biosynthesis were up- regulated in the root, fruit and leaf, except for the MVD gene in the leaf. qRT-PCR further verified the differential expression analysis of selected genes. It shows the highest homology with Sesamum indicum, Handroanthus impetiginosus, Erythranthe guttata, Oleaeuropaea var. sylvestris, and Dorcoceras hygrometricum. A total of 21026 unigenes of transcription factors were assembled into transcription factor families. A total of 8760 unigenes of SSR were assembled. The transcriptome data presented here will make it easier to study the functional genomics of steroidal diosgenin biosynthesis and to change the genes of P. murex to make it more diosgenin.

bioinformatics↗

Analytical Method Development for Evaluation of Pharmacokinetics and Tissue Distribution of Thiourea-Based Antivirals through nLC/MS-MS

Thiourea-based antivirals are organosulfur chemical compounds. Due to their various medicinal uses, including antiviral, antioxidant, and anticancer properties, they are becoming more popular. In this study, pharmacokinetics, Metabolism, bioavailability, and distribution of thiourea derivatives to organs of rats. Thiourea derivatives, namely DSA-00, DSA-02, and DSA-09, exhibit characteristics similar to those of conventional drugs when evaluated in terms of pharmacokinetics, drug-likeness, and medicinal chemistry through in-silico analysis. Then a simple and sensitive analytical method was developed for quantified DSA-00, DSA-02, and DSA-09, using prednisolone as an internal standard (IS) and nano liquid chromatography tandem mass spectrometry (nLC-MS/MS). The plasma and tissue samples were preprocessing with acetonitrile before chromatographic separation by C18 column with isocratic elution using a mobile phase of water-methanol (30:70, v/v) at a flow rate of 0.6 mL/min. A triplequadrupole tandem mass spectrometer in multiple-reaction monitoring (MRM) scanning via an electrospray ionization (ESI) source functioning in negative and positive mode detection. For DSA-00, DSA-02, DSA-09 and, internal standard the optimized mass transition ion-pairs (m/z) for quantification were 297.2, 311.2, 309.09, and 361.19 respectively. Each analytic run required only 15 minutes between injections. The calibration curve for thiourea derivatives showed good linear regression coefficient (R2 >.99) over a range of 1.00-10000 pg/mL except DSA-09. The lower limit of quantification (LLOQ) was 1.00 ng/mL. The intra- and inter-day precisions were no more than 10.8%, and relative errors (RE) ranged from 0.5 to 5.98%. The validated method was effectively used to explore the pharmacokinetics of orally administrative thiourea derivatives in rats.

pharmacology and toxicology↗

Safety Assessment and Evaluation of Novel Thiourea Derivatives as Antivirals

DSA-00 and its two derivatives (DSA-02 & DSA-09) are new thiourea derivatives that exhibit strong antiviral activity against hepatitis B virus comparable to Entecavir. Here these compounds were evaluated for in-vitro cytotoxicity and genotoxicity, and in vivo acute toxicity for their potential therapeutic use. The cytotoxicity of thiourea derivatives was assessed in HepG2 and HepG2.2.15 cells by MTT assay whereas their genotoxicity was measured by Ames II test. The acute toxicity study was carried out in the Sprague-Dawley rats by observing the following parameters: mortality, clinical symptoms, hematological parameters, urine and changes in animal body & organs weight, gross necropsy and histopathology. DSA-00, DSA-02, and DSA-09 were non-cytotoxic even at the 320M concentration with respective CC50 values of 329.6, 323.5, and 349.7 M. The Ames II test revealed that these molecules were non-mutagenic at a [~]1M concentration. The acute toxicity studies revealed LD50 values belong from the moderate range of toxicity of DSA-00, DSA-02 and DSA-09. Importantly, there were no abnormal change in body weight and organs weight. Moreover, no abnormal clinical signs such as hematological parameter, urinalysis, gross necropsy and or histopathological in any of the animals after receiving oral doses of thiourea derivatives. Thiourea derivatives DSA-00, DSA-02, and DSA-09 appear to have a moderate range of acute toxicity at dosages used and thus, appear to be generally safe. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=125 SRC="FIGDIR/small/546447v1_ufig1.gif" ALT="Figure 1"> View larger version (32K): org.highwire.dtl.DTLVardef@8c8abaorg.highwire.dtl.DTLVardef@e7bcb1org.highwire.dtl.DTLVardef@15446dorg.highwire.dtl.DTLVardef@3f5d7b_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGraphical AbstractC_FLOATNO Illustration of the study design and the findings for each aspect of the study: in-vitro cytotoxicity (1), genotoxicity by the Ames test (2), and oral toxicity in rats (3). C_FIG

pharmacology and toxicology↗

Illumina-based Whole De Novo Transcriptome Profiling and Diosgenin Biosynthetic pathway of Tribulus terrestris: A Medicinal herb

Tribulus terrestris L. (Chota gokhuru), an annual herb of the Zygophyllaceae family accumulates steroidal-type triterpenoids of diosgenin in root, leaf, fruit, which have been used as a traditional medicine to cure steroidal hormonal diseases such as estrogen, progesterone, and cardiovascular and urogenital. However, to date, the diosgenin biosynthetic pathway of T. terrestris is unknown. In this research paper, identification of candidate genes involved in bioactive compounds (diosgenin) was conducted via whole transcriptome profiling of T. terrestris using the Illumina HiSeq sequencing platform, which generated 482 million reads. RNA-Seq reads were assembled and clustered into 148871 unigenes. NCBI non-redundant (NR) and Swiss-Prot databases search anchored 50% unigenes with the functional annotations based on similarities of sequence. Further, in Gene Ontology and KEGG-KAAS pathways, a total of 126 unigenes were highly expressed in the terpenoids backbone biosynthesis, in which diosgenin pathways (two genes of glycolytic, three genes of mevalonate, two genes of MEP/DOX, one gene of mono-terpenoids, and seven genes of steroidal pathways) were identified. In addition, 9 genes were randomly selected for confirmation through real-time PCR. T. terrestris showed the highest homology with the Citrus sinensis and Juglans regia. 27478 unigenes of transcription factors were assembled, in which top ten transcription factors were highly expressed. A total of 17623 SSR was assembled, amongst them unigenes of di-nucleotide were highly repeated. This research work will improve our understanding of the diosgenin pathway and lay the groundwork for future molecular research on metabolic engineering, as well as raise diosgenin content in T. terrestris.

molecular biology↗

Natural genetic variation underlying tiller development in barley (Hordeum vulgare L)

In barley (Hordeum vulgare L.), lateral branches called tillers contribute to grain yield and define shoot architecture, but genetic control of tiller number and developmental rate are not well characterized. The primary objectives of this work were to examine relationships between tiller number and other agronomic and morphological traits and identify natural genetic variation associated with tiller number and rate, and related traits. We grew 768 lines from the USDA National Small Grain Core Collection in the field and collected data over two years for tiller number and rate, and agronomic and morphological traits. Our results confirmed that spike row-type and days to heading are correlated with tiller number, and as much as 28% of tiller number variance is attributed to these traits. In addition, negative correlations between tiller number and leaf width and stem diameter were observed, indicating trade-offs between tiller development and other vegetative growth. Thirty-three quantitative trait loci (QTL) were associated with tiller number or rate. Of these, 40% overlapped QTL associated with days to heading and 22% overlapped QTL associated with spike row-type, further supporting that tiller development is influenced by these traits. Despite this, some QTL associated with tiller number or rate, including the major QTL on chromosome 3H, were not associated with any other traits, suggesting that tiller number can be modified independently of other important agronomic traits. These results enhance our knowledge of the genetic control of tiller development in barley, which is important for optimizing tiller number and rate for yield improvement.

genetics↗

Mycobacterium tuberculosis reactivates HIV via exosomes mediated resetting of cellular redox potential and bioenergetics

The synergy between Mycobacterium tuberculosis (Mtb) and HIV-1 interferes with therapy and facilitates pathogenesis of both human pathogens. Fundamental mechanisms by which Mtb exacerbates HIV-1 are not clear. Here, we show that exosomes secreted by macrophages infected with Mtb, including drug-resistant clinical strains, reactivate HIV-1 by inducing oxidative stress. Mechanistically, Mtb-specific exosomes realign mitochondrial and non-mitochondrial oxygen consumption rate (OCR) and modulates the expression of genes mediating oxidative stress response, inflammation, and HIV-1 transactivation. Proteomics revealed the enrichment of several host factors (e.g., HIF-1, galectins, Hsp90) known to promote HIV-1 reactivation in the Mtb-specific exosomes. Treatment with a known antioxidant, N-acetyl cysteine, or with the inhibitors of host factors galectins and Hsp90 attenuated HIV-1 reactivation by Mtb-specific exosomes. Our findings uncovered new paradigms for understanding the redox and bioenergetics basis of HIV-TB co-infection, which will enable the design of effective therapeutic strategies.

microbiology↗

Sequence based mapping identifies AWNS1, a candidate transcription repressor underlying awn suppression at the B1 locus in wheat

Awns are stiff, hair-like structures that grow from the spikelets of wheat (Triticum aestivum L.) and other grasses. In wild wheat relatives, awns play a role in seed dispersal. Awn suppression in domesticated wheat, where awns are shortened or even eliminated entirely, is variably adaptive with both awned and awnless types under widespread cultivation. Although the B1 locus on the long arm of chromosome 5A is a major determinant of awn suppression, no underlying gene or mechanism of action has been identified. Using association mapping, we identified a SNP marker located on the distal end of 5AL (5A28417) predictive of awn status in a panel of 640 U.S. winter wheat breeding lines, indicating that B1 was the determinant of awn suppression in this germplasm. Analysis of historical data available for the panel determined 5A28417 was also significantly associated with grain test weight. Evaluation of spike morphology and kernel traits was undertaken in a 341 RIL population developed from a cross between awned soft winter wheat cultivar LA95135 and awnless cultivar SS-MPV57. Awn suppression in the population co-segregated with 5A28417 and was co-located with QTL for number of spikelets per spike, kernel weight and kernel length. Fine-mapping located B1 to a region containing only two predicted genes, including a C2H2 zinc finger transcription factor 219 bp from 5A28417 that we named AWNS-A1. Deletions encompassing both genes were present in awned mutants of the awnless cultivar Brundage. Polymorphisms in the AWNS-A1 coding region were not observed in diverse wheat germplasm. However, sequencing of wheat lines representing different marker haplotypes in the surrounding region identified a deletion 3 kb downstream of AWNS-A1. A marker for this deletion was highly predictive of awn suppression in a collection of diverse wheat accessions, and indicates that AWNS-A1 is likely the major determinant of awn suppression in global wheat germplasm. AWNS-A1 is more highly expressed in developing spikes of awnless individuals, suggesting a mechanism for awn suppression.

genetics↗

Genome-wide association studies for yield-related traits in soft red winter wheat grown in Virginia

Grain yield is a trait of paramount importance in the breeding of all cereals. In wheat (Triticum aestivum L.), yield has steadily increased since the Green Revolution, though the current rate of increase is not forecasted to keep pace with demand due to growing world population and affluence. While several genome-wide association studies (GWAS) on yield and related component traits have been performed in wheat, the previous lack of a reference genome has made comparisons between studies difficult. In this study, a GWAS for yield and yield-related traits was carried out on a population of 324 soft red winter wheat lines across a total of four rain-fed environments in the state of Virginia using single-nucleotide polymorphism (SNP) marker data generated by a genotyping-by-sequencing (GBS) protocol. Two separate mixed linear models were used to identify significant marker-trait associations (MTAs). The first was a single-locus model utilizing a leave-one-chromosome-out approach to estimating kinship. The second was a sub-setting kinship multi-locus method (FarmCPU). The single-locus model identified nine significant MTAs for various yield-related traits, while the FarmCPU model identified 74 significant MTAs. The availability of the wheat reference genome allowed for the description of MTAs in terms of both genetic and physical positions, and enabled more extensive post-GWAS characterization of significant MTAs. The results indicate promising avenues for increasing grain yield by exploiting variation in traits relating to the number of grains per unit area, as well as phenological traits influencing grain-filling duration of genotypes.

plant biology↗