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Abstract In this study, the endophytic fungus Coniothyrium chaingmaiense-KUMBMDBT-25 was isolated from the healthy stem of Euphorbia tirucalli, mass cultivated by submerged fermentation, and extracted using ethyl acetate as a solvent. The extract was subjected to GC-MS analysis. The synthesized Con-AgNPs were characterized through various bioanalytical methods. The synthesis was confirmed by Bio- spectrophotometry, which showed an absorption peak at 404 nm. FTIR analysis verified the reduction and capping of Con-AgNPs, displaying peaks corresponding to various functional groups. SEM-EDAX and HR-TEM examinations revealed that the Con-AgNPs were spherical, and EDAX analysis confirmed the presence of silver atoms at 3 keV. XRD studies revealed the crystalline structure of Con-AgNPs. DLS and Zeta potential tests determined the size and stability of the synthesized Con-AgNPs, which were 65.81 nm. The Con-AgNPs demonstrated strong antibacterial activity against P. aeruginosa (14.06 ± 0.11 mm, 10 mg/mL) and effective antifungal activity against A. flavus (13.03 ± 0.05 mm, 10 mg/mL). Con-AgNPs exhibited notable biological attributes, including a cytotoxic effect of up to 38.82% and 19.15% at 200 µg/mL in an MTT assay measuring cell viability. Additionally, the nanoparticles demonstrated significant anti-inflammatory effects in both in vitro and in vivo studies, validating the biological and pharmacological potential of Con-AgNPs.

Abstract Hybrid nanofluids’ enhanced thermophysical properties make them applicable in a plethora of mechanical and engineering applications requiring augmented heat transfer. The present study focuses on a three-dimensional Copper-Aluminium Oxide $$\left( Cu\text{- }Al_{2}O_{3}\right)$$ C u - A l 2 O 3 -water based hybrid nanofluid flow within the boundary layer with heat transfer over a rotating exponentially stretching plate, subjected to an inclined magnetic field. The sheet rotates at an angular velocity $$\Omega$$ Ω and the angle of inclination of the magnetic field is $$\gamma$$ γ . Employing a set of appropriate similarity transformation reduces the governing PDEs to ODEs. The resulting ODEs are solved with the finite difference code with Shooting Technique. Primary velocity increases at large rotation but the secondary velocity reduces as the rotation increases. In addition, the magnetic field is found to oppose the flow and thereby causing a reduction in both the primary and secondary velocities. Increasing the volume fraction reduces the skin friction coefficient and enhances the heat transfer rate.

Abstract Epitranscriptome modifications are crucial in translation regulation and essential for maintaining cellular homeostasis. N6 methyladenosine (m6A) is one of the most abundant and well-conserved epitranscriptome modifications, which is known to play a pivotal role in diverse aspects of neuronal functions. However, the role of m6A modifications with respect to activity-mediated translation regulation and synaptic plasticity has not been studied. Here, we investigated the role of m6A modification in response to NMDAR stimulation. We have consistently observed that 5 min NMDAR stimulation causes an increase in eEF2 phosphorylation. Correspondingly, NMDAR stimulation caused a significant increase in the m6A signal at 5 min time point, correlating with the global translation inhibition. The NMDAR induced increase in the m6A signal is accompanied by the redistribution of the m6A marked RNAs from translating to the non-translating pool of ribosomes. The increased m6A levels are well correlated with the reduced FTO levels observed on NMDAR stimulation. Additionally, we show that inhibition of FTO prevents NMDAR mediated changes in m6A levels. Overall, our results establish RNA-based molecular readout which corelates with the NMDAR-dependent translation regulation which helps in understanding changes in protein synthesis.

Abstract Influenza remains a world-wide health concern, causing 290,000–600,000 deaths and up to 5 million cases of severe illnesses annually. Noticing the host factors that control biological responses, such as inflammatory cytokine secretion, to influenza virus infection is important for the development of novel drugs. Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid metabolite and has essential biological functions in inflammation. However, the kinetic effects of influenza virus infection on physiological S1P levels and their signaling in multiple tissues remain unknown. In this study, we utilized a mouse model intranasally infected with 50 or 500 plaque forming units (PFU) of A/Puerto Rico/8/34 (H1N1; PR8) virus to investigate how S1P levels and expression of its regulating factors are affected by influenza virus infection by the liquid-chromatography/mass spectrometry and real-time PCR, respectively. The S1P level was significantly high in the plasma of mice infected with 500 PFU of the virus than that in control mice at 6 day-post-infection (dpi). Elevated gene expression of sphingosine kinase-1 (Sphk1), an S1P synthase, was observed in the liver, lung, white adipose tissue, heart, and aorta of infected mice. This could be responsible for the increased plasma S1P levels as well as the decrease in the hepatic S1P lyase (Sgpl1) gene in the infected mice. These results indicate modulation of S1P-signaling by influenza virus infection. Since S1P regulates inflammation and leukocyte migration, it must be worth trying to target this signaling to control influenza-associated symptoms.

Abstract The role of RORγ as a transcription factor for Th17 cell differentiation and thereby regulation of IL-17 levels is well known. Increased RORγ expression along with IL-17A levels was observed in animal models, immune cells and BAL fluid of COPD patients. Increased IL-17A levels in severe COPD patients are positively correlated with decreased lung functions and increased severity symptoms and emphysema, supporting an urgency to develop novel therapies modulating IL-17 or RORγ for COPD treatment. We identified a potent RORγ inhibitor, PCCR-1 using hit to lead identification followed by extensive lead optimization by structure–activity relationship. PCCR-1 resulted in RORγ inhibition with a high degree of specificity in a biochemical assay, with > 300-fold selectivity over other isoforms of ROR. Our data suggest promising potency for IL-17A inhibition in human and canine PBMCs and mouse splenocytes with no significant impact on Th1 and Th2 cytokines. In vivo, PCCR-1 exhibited significant efficacy in the acute CS model with dose-dependent inhibition of the PD biomarkers that correlated well with the drug concentration in lung and BAL fluid, demonstrating an acceptable safety profile. This inhibitor effectively inhibited IL-17A release in whole blood and BALf samples from COPD patients. Overall, we identified a selective inhibitor of RORγ to pursue further development of novel scaffolds for COPD treatment.

Carcinogenic effect of tobacco in oral cancer is through chewing and/or smoking. Significant differences exist in development of oral cancer between tobacco users and non-users. However, molecular alterations induced by different forms of tobacco are yet to be fully elucidated. We developed cellular models of chronic exposure to chewing tobacco and cigarette smoke using immortalized oral keratinocytes. Chronic exposure to tobacco resulted in increased cell scattering and invasiveness in immortalized oral keratinocytes. miRNA sequencing using Illumina HiSeq 2500 resulted in the identification of 10 significantly dysregulated miRNAs (4 fold; p ≤ 0.05) in chewing tobacco treated cells and 6 in cigarette smoke exposed cells. We integrated this data with global proteomic data and identified 36 protein targets that showed inverse expression pattern in chewing tobacco treated cells and 16 protein targets that showed inverse expression in smoke exposed cells. In addition, we identified 6 novel miRNAs in chewing tobacco treated cells and 18 novel miRNAs in smoke exposed cells. Integrative analysis of dysregulated miRNAs and their targets indicates that signaling mechanisms leading to oncogenic transformation are distinct between both forms of tobacco. Our study demonstrates alterations in miRNA expression in oral cells in response to two frequently used forms of tobacco.

Abstract In-car passive stress sensing could enable the monitoring of stress biomarkers while driving and reach millions of commuters daily (i.e., 123 million daily commuters in the US alone). Here, we present a nonintrusive method to detect stress solely from steering angle data of a regular car. The method uses inverse filtering to convert angular movement data into a biomechanical Mass Spring Damper model of the arm and extracts its damped natural frequency as an approximation of muscle stiffness, which in turn reflects stress. We ran a within-subject study (N = 22), in which commuters drove a vehicle around a closed circuit in both stress and calm conditions. As hypothesized, cohort analysis revealed a significantly higher damped natural frequency for the stress condition (P = .023, d = 0.723). Subsequent automation of the method achieved rapid (i.e., within 8 turns) stress detection in the individual with a detection accuracy of 77%.

Abstract Human error has been implicated as a causal factor in a large proportion of road accidents. Automated driving systems purport to mitigate this risk, but self-driving systems that allow a driver to entirely disengage from the driving task also require the driver to monitor the environment and take control when necessary. Given that sleep loss impairs monitoring performance and there is a high prevalence of sleep deficiency in modern society, we hypothesized that supervising a self-driving vehicle would unmask latent sleepiness compared to manually controlled driving among individuals following their typical sleep schedules. We found that participants felt sleepier, had more involuntary transitions to sleep, had slower reaction times and more attentional failures, and showed substantial modifications in brain synchronization during and following an autonomous drive compared to a manually controlled drive. Our findings suggest that the introduction of partial self-driving capabilities in vehicles has the potential to paradoxically increase accident risk.

Abstract In this study, a mathematical model is developed to scrutinize the transient magnetic flow of Cross nanoliquid past a stretching sheet with thermal radiation effects. Binary chemical reactions and heat source/sink effects along with convective boundary condition are also taken into the consideration. Appropriate similarity transformations are utilized to transform partial differential equations (PDE’s) into ordinary ones and then numerically tackled by shooting method. The impacts of different emerging parameters on the thermal, concentration, velocity, and micro-rotation profiles are incorporated and discussed in detail by means of graphs. Results reveal that, the escalation in magnetic parameter and Rayleigh number slowdowns the velocity and momentum of the fluid. The increase in Biot number, radiation and heat sink/source parameters upsurges the thermal boundary but, converse trend is seen for escalating Prandtl number. The density number of motile microorganisms acts as a growing function of bioconvection Lewis number and declining function of bioconvection Peclet number.

Abstract To estimate the reproductive number (R0) of the coronavirus in the present scenario and to predict the incidence of daily and probable cumulative cases, by 20 August, 2020 for Karnataka state in India. The model used serial interval with a gamma distribution and applied ‘early R’ to estimate the R0 and ‘projections’ package in R program. This was performed to mimic the probable cumulative epidemic trajectories and predict future daily incidence by fitting the data to existing daily incidence and the estimated R0 by a model based on the assumption that daily incidence follows Poisson distribution. The maximum-likelihood (ML) value of R0 was 2.242 for COVID-19 outbreak, as on June 2020. The median with 95% CI of R0 values was 2.242 (1.50–3.00) estimated by bootstrap resampling method. The expected number of new cases for the next 60 days would progressively increase, and the estimated cumulative cases would reach 27,238 (26,008–28,467) at the end of 60th day in the future. But, if R0 value was doubled the estimated total number of cumulative cases would increase up to 432,411 (400,929–463,893) and if, R0 increase by 50%, the cases would increase up to 86,386 (80,910–91,861). The probable outbreak size and future daily cumulative incidence are largely dependent on the change in R0 values. Hence, it is vital to expedite the hospital provisions, medical facility enhancement work, and number of random tests for COVID-19 at a very rapid pace to prepare the state for exponential growth in next 2 months.

Abstract Shammah is a smokeless tobacco product often mixed with lime, ash, black pepper and flavorings. Exposure to shammah has been linked with dental diseases and oral squamous cell carcinoma. There is limited literature on the prevalence of shammah and its role in pathobiology of oral cancer. In this study, we developed a cellular model to understand the effect of chronic shammah exposure on oral keratinocytes. Chronic exposure to shammah resulted in increased proliferation and invasiveness of non-transformed oral keratinocytes. Quantitative proteomics of shammah treated cells compared to untreated cells led to quantification of 4712 proteins of which 402 were found to be significantly altered. In addition, phosphoproteomics analysis of shammah treated cells compared to untreated revealed hyperphosphorylation of 36 proteins and hypophosphorylation of 83 proteins (twofold, p-value ≤ 0.05). Bioinformatics analysis of significantly altered proteins showed enrichment of proteins involved in extracellular matrix interactions, necroptosis and peroxisome mediated fatty acid oxidation. Kinase-Substrate Enrichment Analysis showed significant increase in activity of kinases such as ROCK1, RAF1, PRKCE and HIPK2 in shammah treated cells. These results provide better understanding of how shammah transforms non-neoplastic cells and warrants additional studies that may assist in improved early diagnosis and treatment of shammah induced oral cancer.

Abstract Oral squamous cell carcinoma (OSCC) is known for its aggressiveness associated with poor prognosis. The molecular mechanisms underlying the invasion and metastasis are still poorly understood. An improved understanding of these mechanisms shall precede the development of new diagnostic tools and targeted therapies. We report an integrated approach using bioinformatics to predict candidate genes, coupled with proteomics and immunohistochemistry for validating their presence and involvement in OSCC pathways heralding invasion and metastasis. Four genes POSTN, TNC, CAV1 and FSCN1 were identified. A protein–protein interaction network analysis teamed with pathway analysis led us to propose the role of the identified genes in invasion and metastasis in OSCC. Further analyses of archived FFPE blocks of various grades of oral cancer was carried out using TMT-based mass spectrometry and immunohistochemistry. Results of this study expressed a strong communiqué and interrelationship between these candidate genes. This study emphasizes the significance of a molecular biomarker panel as a diagnostic tool and its correlation with the invasion and metastatic pathway of OSCC. An insight into the probable association of CAF's and these biomarkers in the evolution and malignant transformation of OSCC further magnifies the molecular-biological spectrum of OSCC tumour microenvironment.

Abstract Docosahexaenoic acid (DHA) is known to inhibit breast cancer in the rat. Here we investigated whether DHA itself or select metabolites can account for its antitumor action. We focused on metabolites derived from the lipoxygenase (LOX) pathway since we previously showed that they were superior anti-proliferating agents compared to DHA; 4-OXO-DHA was the most potent. A lipidomics approach detected several LOX-metabolites in plasma and the mammary gland in rats fed DHA; we also identified for the first time, 4-OXO-DHA in rat plasma. In a reporter assay, 4-OXO-DHA and 4-HDHA were more effective activators of PPARɣ than DHA. In breast cancer cell lines, 4-OXO-DHA induced PPARɣ and 15-hydroxyprostaglandin dehydrogenase (15-PGDH) but inhibited the activity of NF-κB and suppressed PI3K and mTOR signaling. Because of the structural characteristics of 4-OXO-DHA (Michael acceptor), not shared by any of the other hydroxylated-DHA, we used MS and showed that it can covalently modify the cysteine residue of NF-κB. We have also shown that the chemopreventive effect of DHA is associated with significant reduction of PGE2 levels, in both rat mammary tumors induced by MNU and non-involved mammary tissues. Collectively, our results indicate that 4-OXO-DHA is the metabolite of choice in future chemoprevention studies.

Abstract Bacterial blight caused by Xanthomonas axonopodis pv. punicae is a major disease of pomegranate. Bacterial blight drastically reduces the yield and quality of fruits, which are critical for pomegranate production. Precise and early diagnosis of bacterial blight is crucial for active surveillance and effective management of the disease. Symptoms based disease diagnostic methods are labor-intensive, time-consuming and may not detect disease on asymptomatic plants. DNA-based disease diagnostics using polymerase chain reaction (PCR) are reliable, precise, accurate and quick. PCR coupled with agarose gel electrophoresis (PCR-AGE), PCR coupled with capillary electrophoresis (PCR-CE) and real-time PCR (qPCR) were applied for the early and accurate diagnosis of bacterial blight in pomegranate. PCR-CE and qPCR were capable of diagnosing bacterial blight 6 to 10 days before symptom appearance, with detection limits of 100 fg and 10 fg of bacterial DNA respectively. However, conventional PCR-AGE detected pathogen at the onset of disease symptoms with a detection limit of 10 pg of bacterial DNA. qPCR detected bacterial blight in orchards that did not show any disease symptoms. Our data demonstrate that qPCR is more sensitive than other PCR methods along with being reliable for early diagnosis.

Abstract The use of Bt proteins in crops has revolutionized insect pest management by offering effective season-long control. However, field-evolved resistance to Bt proteins threatens their utility and durability. A recent example is field-evolved resistance to Cry1Fa and Cry1A.105 in fall armyworm (Spodoptera frugiperda). This resistance has been detected in Puerto Rico, mainland USA, and Brazil. A S. frugiperda population with suspected resistance to Cry1Fa was sampled from a maize field in Puerto Rico and used to develop a resistant lab colony. The colony demonstrated resistance to Cry1Fa and partial cross-resistance to Cry1A.105 in diet bioassays. Using genetic crosses and proteomics, we show that this resistance is due to loss-of-function mutations in the ABCC2 gene. We characterize two novel mutant alleles from Puerto Rico. We also find that these alleles are absent in a broad screen of partially resistant Brazilian populations. These findings confirm that ABCC2 is a receptor for Cry1Fa and Cry1A.105 in S. frugiperda, and lay the groundwork for genetically enabled resistance management in this species, with the caution that there may be several distinct ABCC2 resistances alleles in nature.

Abstract Carcinogenic effect of tobacco in oral cancer is through chewing and/or smoking. Significant differences exist in development of oral cancer between tobacco users and non-users. However, molecular alterations induced by different forms of tobacco are yet to be fully elucidated. We developed cellular models of chronic exposure to chewing tobacco and cigarette smoke using immortalized oral keratinocytes. Chronic exposure to tobacco resulted in increased cell scattering and invasiveness in immortalized oral keratinocytes. miRNA sequencing using Illumina HiSeq 2500 resulted in the identification of 10 significantly dysregulated miRNAs (4 fold; p ≤ 0.05) in chewing tobacco treated cells and 6 in cigarette smoke exposed cells. We integrated this data with global proteomic data and identified 36 protein targets that showed inverse expression pattern in chewing tobacco treated cells and 16 protein targets that showed inverse expression in smoke exposed cells. In addition, we identified 6 novel miRNAs in chewing tobacco treated cells and 18 novel miRNAs in smoke exposed cells. Integrative analysis of dysregulated miRNAs and their targets indicates that signaling mechanisms leading to oncogenic transformation are distinct between both forms of tobacco. Our study demonstrates alterations in miRNA expression in oral cells in response to two frequently used forms of tobacco.