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The creation of ex vivohuman liver models has long been a critical objective in academic, clinical, and pharmaceutical research, particularly for drug development, where accurate evaluation of hepatic metabolic dynamics is crucial. We have developed a bioengineered, perfused, organ-level human liver model that accurately replicates key liver functions, including metabolic activities, and protein synthesis, thus addressing some of the limitations associated with traditional liver monolayers, organoids, and matrix-embedded liver cells. Our approach utilizes liver-specific biomatrix scaffolds, prepared using an innovative protocol and fortified with matrix components that facilitate cellular interactions. These scaffolds, when seeded with human fetal liver cells or co-seeded with liver parenchymal and endothelial cell lines, enable the formation of three-dimensional (3D) human livers with enhanced cellular organization. The “recellularized tissue-engineered livers” (RCLs) have undergone various analyses, demonstrating the capability for establishing liver microenvironments ex vivo. Within 7–14 days, the RCLs exhibit evidence of liver differentiation and metabolic capabilities, underscoring the potential for use in drug metabolism and toxicity studies. Although our study represents a significant step forward, we acknowledge the need for direct comparisons with existing models and further research to fully elucidate the spectrum of regenerative responses. The high drug-metabolizing enzyme activity of RCLs, as demonstrated in our study, provides a promising avenue for investigating drug-induced liver injury mechanisms, contributing to a more detailed understanding of early drug discovery processes.

Biomethane production from lignocellulosic biomass via anaerobic digestion (AD) is a promising avenue for bioenergy. However, maximizing biomethane yield using developed microbial consortium (DMC) remains a complex challenge. This study aims to elucidate the comparative efficacy of a purposefully DMC under controlled conditions for AD of kallar grass (KG), rice husk (RH) and wheat residue (WR). The DMC encompasses Bacteroidetes, Firmicutes, Proteobacteria, Euryarchaeota, and Chloroflexi, with methanogens representing 14.6% of the community. Thorough physico-chemical characteristics and ultimate analyses provided comprehensive insights into compositions and bio-methane potentials prediction of biomasses. The digester revealed higher biomethane potentials (BMPs) with short lag phase and retention time and showed methane content of 61–64.6% during the first week of AD. The amount of volatile fatty acids (VFA) did not exceed the threshold levels, which facilitated the smooth operation of AD. The BMP was found highest in KG (289.7 mL/g VS), followed by RH (283.3 mL/g VS), and WR(269.7 mL/g VS) which is significantly higher than previously reported studies. The modified Gompertz model showed the best fit, followed by logistic and transference function models. The observed results signpost the tremendous potential of waste biomass particularly KG, RH and WRto produce biomethane by DMC. This study provides crucial insights for optimizing BMP emphasizing the promising prospects of KG, RH, and WRfor sustainable bioenergy applications.

Herein, the electrochemical technique was employed to detect hydroquinone (HQ) using a modified glassy carbon electrode (GCE) with reduced graphene oxide (rGO) and silver (Ag)-decorated tin oxy-nanoparticles (SnONPs) to form Ag@SnONPs/rGO nanocomposites (NC). The Ag@SnONPs/rGO nanocomposites were morphologically characterized using multiple analytical methods such as XRD, Raman, XPS, HR-SEM, and HR-TEM. This study revealed that Ag@SnONPs/rGO-NC exhibits excellent conductivity due to the presence of rGO that provides potential π–π interactions with SnONPs, while Ag enhances electron-transfer kinetics. This facilitates efficient charge transport within the sensor, thereby improving HQ adsorption. The key advantages of the sensor demonstrate a concentration of 0.5–200 µM, and a low detection limit value of 0.010 µM, and a high sensitivity value of 6.0746 µA µM−1cm2. Under optimal conditions, the Ag@SnONPs/rGO sensor may be used to determine HQ and its concentration using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The Ag@SnONPs-rGO/GCE sensor demonstrated excellent reproducibility, repeatability, and stability. Moreover, the suggested bimetallic nanocomposite effectively determined the presence of HQ in water and cosmetic samples.

The phenomenon of linear motion of conductor in a magnetic field is commonly found in electric machineries, such as electromagnetic brakes, linear induction motor, electromagnetic flowmeter, and so on. The design and analysis of the same require an accurate evaluation of induced currents and the associated reaction magnetic fields. The finite-element method (FEM) is a generally employed numerical technique for this purpose. However, it needs stabilization techniques to provide an accurate solution. In this work, such a stabilization technique is developed for the edge elements. The stability and, hence, the accuracy are brought in by a suitable representation of the source term. The stability and accuracy of the proposed scheme are first shown analytically and then demonstrated with the help of 2-D and 3-D simulations. The proposed scheme would require a graded regular mesh along the direction of motion.

Adaptive mesh refinement (AMR) is the art of solving PDEs on a mesh hierarchy with increasing mesh refinement at each level of the hierarchy. Accurate treatment on AMR hierarchies requires accurate prolongation of the solution from a coarse mesh to a newly defined finer mesh. For scalar variables, suitably high-order finite volume WENO methods can carry out such a prolongation. However, classes of PDEs, such as computational electrodynamics (CED) and magnetohydrodynamics (MHD), require that vector fields preserve a divergence constraint. The primal variables in such schemes consist of normal components of the vector field that are collocated at the faces of the mesh. As a result, the reconstruction and prolongation strategies for divergence constraint-preserving vector fields are necessarily more intricate. In this paper we present a fourth-order divergence constraint-preserving prolongation strategy that is analytically exact. Extension to higher orders using analytically exact methods is very challenging. To overcome that challenge, a novel WENO-like reconstruction strategy is invented that matches the moments of the vector field in the faces, where the vector field components are collocated. This approach is almost divergence constraint-preserving, therefore, we call it WENO-ADP. To make it exactly divergence constraint-preserving, a touch-up procedure is developed that is based on a constrained least squares (CLSQ) method for restoring the divergence constraint up to machine accuracy. With the touch-up, it is called WENO-ADPT. It is shown that refinement ratios of two and higher can be accommodated. An item of broader interest in this work is that we have also been able to invent very efficient finite volumeWENO methods, where the coefficients are very easily obtained and the multidimensional smoothness indicators can be expressed as perfect squares. We demonstrate that the divergence constraint-preserving strategy works at several high orders for divergence-free vector fields as well as vector fields, where the divergence of the vector field has to match a charge density and its higher moments. We also show that our methods overcome the late time instability that has been known to plague adaptive computations in CED.

GPU computing is expected to play an integral part in all modern Exascale supercomputers. It is also expected that higher order Godunov schemes will make up about a significant fraction of the application mix on such supercomputers. It is, therefore, very important to prepare the community of users of higher order schemes for hyperbolic PDEs for this emerging opportunity.

Fibrolamellar carcinoma (FLC) is a rare, lethal, early-onset liver cancer with a critical need for new therapeutics. The primary driver in FLC is the fusion oncoprotein, DNAJ-PKAc, which remains challenging to target therapeutically. It is critical, therefore, to expand understanding of the FLC molecular landscape to identify druggable pathways/targets. Here, we perform the most comprehensive integrative proteo-metabolomic analysis of FLC. We also conduct nutrient manipulation, respirometry analyses, as well as key loss-of-function assays in FLC tumor tissue slices from patients. We propose a model of cellular energetics in FLC pointing to proline anabolism being mediated by ornithine aminotransferase hyperactivity and ornithine transcarbamylase hypoactivity with serine and glutamine catabolism fueling the process. We highlight FLC’s potential dependency on voltage-dependent anion channel (VDAC), a mitochondrial gatekeeper for anions including pyruvate. The metabolic rewiring in FLC that we propose in our model, with an emphasis on mitochondria, can be exploited for therapeutic vulnerabilities.

To identify pain-relevant miRNAs downstream of beta adrenergic receptor (βAR) activation and determine their functional effects in chronic primary pain conditions (CPPCs). Rats received peripheral delivery of the β2- and β3AR antagonists ICI-118,511 (1.5mg/kg/day) and SR59230A (1.67 mg/kg/day) alongside systemic delivery of the catechol-o-methlytransferase inhibitor OR486 (15mg/kg/day) or vehicle over 14 days (N=20). RNA sequencing shows that OR486 treatment decreased levels of miR-133a in circulation, and this downregulation was prevented by βAR antagonism. miR-133a downregulation was replicated in acute (N=32) and sustained (N=23) CPPC mice and in human cohorts of vestibulodynia (N=75) and fibromyalgia (N=48) patients. qPCR in peripheral (adipose, muscle) and central (spinal cord) tissues collected from CPPC mice indicates that miR-133a was downregulated in adipose at both times, while in the spinal cord there was differential expression only at the 14-day time point. Further, β3AR activation in primary adipocytes drives miR-133a downregulation (N=15). We then induced adipose-specific overexpression of miR-133a to directly test its role in modulating pain. Compared to CPPC mice receiving a nonsense sequence, those treated with adipose-specific miR-133a exhibit significant increases in mechanical pain thresholds at multiple body sites, indicative of widespread analgesia (N=51). Further, miR-133a overexpression in primary dorsal root ganglion cultures had anti-nociceptive effects on capsaicin-induced phosphorylation of extracellular signal-regulated kinase (N=12) and capsaicin-induced calcium responses (N=32). These data provide the first link between miR-133a and CPPCs across species and support the use of peripherally-targeted miR-133a overexpression strategies for the treatment of CPPCs. Funding: NIH/NINDS R03NS106166 and R01NS109541 A Nackley. NIH/NIGMS T32GM133352 and NIH/NINDS F31NS130861 to N Hernandez.

Fibrolamellar carcinoma (FLC) is a liver tumor with a high mortality burden and few treatment options. A promising therapeutic vulnerability in FLC is its driver mutation, a conserved DNAJB1-PRKACAgene fusion that could be an ideal target neoantigen for immunotherapy. In this study, we aim to define endogenous CD8 T cell responses to this fusion in FLC patients and evaluate fusion-specific T cell receptors (TCRs) for use in cellular immunotherapies. We observe that fusion-specific CD8 T cells are rare and that FLC patient TCR repertoires lack large clusters of related TCR sequences characteristic of potent antigen-specific responses, potentially explaining why endogenous immune responses are insufficient to clear FLC tumors. Nevertheless, we define two functional fusion-specific TCRs, one of which has strong anti-tumor activity in vivo. Together, our results provide insights into the fragmented nature of neoantigen-specific repertoires in humans and indicate routes for clinical development of successful immunotherapies for FLC.

In this Resource/Methodology, O'Connor et al. performed high-throughput sequencing of RNA isolated by cross-linking immunoprecipitation (HITS-CLIP) to comprehensively characterize the network of high-confidence, in vivo mRNA:miRNA interactions across white and brown fat, revealing >27,000 unique AGO binding sites. Their work represents the only experimentally generated miRNA targetome in adipose tissue and identifies multiple regulatory pathways that may specify the unique identities of white and brown fat.

Dyslipidemia and insulin resistance are significant adverse outcomes of consuming high-sugar diets. Conversely, dietary fish oil (FO) reduces plasma lipids. Diet-induced dyslipidemia in a rhesus model better approximates the pathophysiology of human metabolic syndrome (MetS) than rodent models. Here, we investigated relationships between metabolic parameters and hypertriglyceridemia in rhesus macaques consuming a high-fructose diet (n = 59) and determined the effects of FO supplementation or RNA interference (RNAi) on plasma ApoC3 and triglyceride (TG) concentrations. Fructose supplementation increased body weight, fasting insulin, leptin, TGs, and large VLDL particles and reduced adiponectin concentrations (all P< 0.001). In multiple regression analyses, increased plasma ApoC3 was the most consistent and significant variable related to diet-induced hypertriglyceridemia. FO supplementation, which attenuated increases of plasma TG and ApoC3 concentrations, reversed fructose-induced shifts of lipoprotein particle size toward IDL and VLDL, a likely mechanism contributing to beneficial metabolic effects, and reduced hepatic expression of genes regulated by the SREBP pathway, particularly acetyl-CoA carboxylase. Furthermore, RNAi-mediated ApoC3 inhibition lowered plasma TG concentrations in animals with diet-induced hypertriglyceridemia. In summary, ApoC3 is an important independent correlate of TG-rich lipoprotein concentrations in rhesus macaques consuming a high-fructose diet. ApoC3 is a promising therapeutic target for hypertriglyceridemia in patients with MetS and diabetes.

Periodontitis is associated with a dysbiotic shift in the oral microbiome. Vaccine approaches to prevent microbial shifts from healthy to diseased state in oral biofilms would provide a fundamental therapeutic strategy against periodontitis. Since dental plaque formation is a polymicrobial and multilayered process, vaccines targeting single bacterial species would have limited efficacy in clinical applications. In this study, we developed a divalent mucosal vaccine consisting of a mixture of FlaB-tFomA and Hgp44-FlaB fusion proteins targeting virulence factors of inflammophilic bacteria Fusobacterium nucleatumand Porphyromonas gingivalis, respectively. Introduction of peptide linkers between FlaB and antigen improved the stability and immunogenicity of engineered vaccine antigens. The intranasal immunization of divalent vaccine induced protective immune responses inhibiting alveolar bone loss elicited by F. nucleatumand P. gingivalisinfection. The built-in flagellin adjuvant fused to protective antigens enhanced antigen-specific antibody responses and class switch recombination. The divalent vaccine antisera recognized natural forms of surface antigens and reacted with diverse clinical isolates of Fusobacteriumsubspecies and P. gingivalis. The antisera inhibited F. nucleatum-mediated biofilm formation, co-aggregation of P. gingivalisand Treponema denticola, and P. gingivalis-host cell interactions. Taken together, the built-in adjuvant-engineered mucosal vaccine provides a technological platform for multivalent periodontitis vaccines targeting dysbiotic microbiome.

Periodontitis is associated with a dysbiotic shift in the oral microbiome. Vaccine approaches to prevent microbial shifts from healthy to diseased state in oral biofilms would provide a fundamental therapeutic strategy against periodontitis. Since dental plaque formation is a polymicrobial and multilayered process, vaccines targeting single bacterial species would have limited efficacy in clinical applications. In this study, we developed a divalent mucosal vaccine consisting of a mixture of FlaB-tFomA and Hgp44-FlaB fusion proteins targeting virulence factors of inflammophilic bacteria Fusobacterium nucleatumand Porphyromonas gingivalis, respectively. Introduction of peptide linkers between FlaB and antigen improved the stability and immunogenicity of engineered vaccine antigens. The intranasal immunization of divalent vaccine induced protective immune responses inhibiting alveolar bone loss elicited by F. nucleatumand P. gingivalisinfection. The built-in flagellin adjuvant fused to protective antigens enhanced antigen-specific antibody responses and class switch recombination. The divalent vaccine antisera recognized natural forms of surface antigens and reacted with diverse clinical isolates of Fusobacteriumsubspecies and P. gingivalis. The antisera inhibited F. nucleatum-mediated biofilm formation, co-aggregation of P. gingivalisand Treponema denticola, and P. gingivalis-host cell interactions. Taken together, the built-in adjuvant-engineered mucosal vaccine provides a technological platform for multivalent periodontitis vaccines targeting dysbiotic microbiome.

Arsenic (As) is a toxic metalloid. Inorganic arsenic (iAs) is a form of As commonly found in drinking water and in some foods. Overwhelming evidence suggests that people chronically exposed to iAs are at risk of developing cancer or cardiovascular, neurological, and metabolic diseases. Although the mechanisms underlying iAs-associated illness remain poorly characterized, a growing body of literature raises the possibility that microRNAs (miRNAs), post-transcriptional gene suppressors, may serve as mediators and/or early indicators of the pathologies associated with iAs exposure. To characterize the circulating miRNA profiles of individuals chronically exposed to iAs, samples of plasma were collected from 109 healthy residents of the city of Zimapán and the Lagunera area in Mexico, the regions with historically high exposures to iAs in drinking water. These plasma samples were analyzed for small RNAs using high-throughput sequencing and for iAs and its methylated metabolites. Associations between plasma levels of arsenic species and miRNAs were evaluated. Six circulating miRNAs (miRs-423-5p, -142-5p -2, -423-5p +1, -320c-1, -320c-2, and -454-5p), two of which have been previously linked to cardiovascular disease and diabetes (miRs-423-5p, -454-5p), were found to be significantly correlated with plasma MAs. No miRNAs were associated with plasma iAs or DMAs after correction for multiple testing. These miRNAs may represent mechanistic links between iAs exposure and disease or serve as markers of disease risks associated with this exposure.

Additive manufacturing, using three-dimensional (3D) printing technology, is considered revolutionary in manufacturing, pharmaceutical, food, and biomedical industries, as it provides a platform for constructing a wide range of tailored object geometries. This rapidly growing technology mainly uses renewable natural polymers combined with synthetic materials to achieve the printability with desired features of the printed 3D structures. Among different polymers, lignin is receiving immense consideration as a renewable bio-based raw material for manufacturing high-performance 3D printed products owing to its amorphous macromolecular structure and antioxidative, antimicrobial, and other beneficial properties. Lignin is a natural organic polymer found in all plants, particularly wood and tree bark, with few exceptions like bryophytes. It accounts for about 30% of the total biomass worldwide, ranking it as the second most abundant renewable material after cellulose. Despite such advantages, the complex polymeric and amorphous structure of lignin is a great challenge to its utilization in 3D printing for the manufacturing of high-value-added products. This review summarizes the use of lignin in the preparation of ink for the 3D printing of different materials, such as degradable composites, hydrogels, and 3D thermoplastic materials. This review discusses the potential merits and limitations of different types of lignin and their innate features in the preparation of 3D printed materials. The challenges and future perspectives for use of lignin in the preparation of ink for 3D printing are also discussed to underscore the critical issues and opportunities.

To date, numerous nucleic acid species have been detected in the systemic circulation including microRNAs (miRNAs); however, their functional role in this compartment remains unclear.

Plasma membrane (PM) free cholesterol (FC) is emerging as an important modulator of signal transduction. Here, we show that hepatocyte-specific knockout (HSKO) of the cellular FC exporter, ATP-binding cassette transporter A1 (ABCA1), leads to decreased PM FC content and defective trafficking of lysosomal FC to the PM. Compared with controls, chow-fed HSKO mice had reduced hepatic (1) insulin-stimulated Akt phosphorylation, (2) activation of the lipogenic transcription factor Sterol Regulatory Element Binding Protein (SREBP)-1c, and (3) lipogenic gene expression. Consequently, Western-type diet-fed HSKO mice were protected from steatosis. Surprisingly, HSKO mice had intact glucose metabolism; they showed normal gluconeogenic gene suppression in response to re-feeding and normal glucose and insulin tolerance. We conclude that: (1) ABCA1 maintains optimal hepatocyte PM FC, through intracellular FC trafficking, for efficient insulin signaling; and (2) hepatocyte ABCA1 deletion produces a form of selective insulin resistance so that lipogenesis is suppressed but glucose metabolism remains normal.

Maintenance of genomic integrity is critical during neurodevelopment, particularly in rapidly dividing cerebellar granule neuronal precursors that experience constitutive replication-associated DNA damage. As Dicer was recently recognized to have an unexpected function in the DNA damage response, we examined whether Dicer was important for preserving genomic integrity in the developing brain. We report that deletion of Dicer in the developing mouse cerebellum resulted in the accumulation of DNA damage leading to cerebellar progenitor degeneration, which was rescued with p53 deficiency; deletion of DGCR8 also resulted in similar DNA damage and cerebellar degeneration. Dicer deficiency also resulted in DNA damage and death in other rapidly dividing cells including embryonic stem cells and the malignant cerebellar progenitors in a mouse model of medulloblastoma. Together, these results identify an essential function of Dicer in resolving the spontaneous DNA damage that occurs during the rapid proliferation of developmental progenitors and malignant cells.

Estrogen receptor α (ERα) is an important biomarker of breast cancer severity and a common therapeutic target. In response to estrogen, ERα stimulates a dynamic transcriptional program including both coding and noncoding RNAs. We generate a fine-scale map of expression dynamics by performing a temporal profiling of both messenger RNAs (mRNAs) and microRNAs (miRNAs) in MCF-7 cells (an ER+ model cell line for breast cancer) in response to estrogen stimulation. We identified three primary expression trends—transient, induced, and repressed—that were each enriched for genes with distinct cellular functions. Integrative analysis of mRNA and miRNA temporal expression profiles identified miR-503 as the strongest candidate master regulator of the estrogen response, in part through suppression of ZNF217—an oncogene that is frequently amplified in cancer. We confirmed experimentally that miR-503 directly targets ZNF217and that overexpression of miR-503 suppresses MCF-7 cell proliferation. Moreover, the levels of ZNF217and miR-503 are associated with opposite outcomes in breast cancer patient cohorts, with high expression of ZNF217associated with poor survival and high expression of miR-503 associated with improved survival. Overall, these data indicate that miR-503 acts as a potent estrogen-induced candidate tumor suppressor miRNA that opposes cellular proliferation and has promise as a novel therapeutic for breast cancer. More generally, our work provides a systems-level framework for identifying functional interactions that shape the temporal dynamics of gene expression.

MicroRNAs (miRNAs) are small noncoding RNA molecules that modulate gene expression at the posttranscriptional level. Numerous reports have elucidated the importance of miRNAs in the regulation of a wide array of biological processes including metabolism and energy homeostasis. miRNAs in the endocrine pancreas have been intensively studied over the last 15 years and linked to pancreatic islet development and function. In comparison, knowledge of miRNAs in gut endocrine cells, or enteroendocrine cells (EECs), is severely lacking. EECs have important roles in systemic energy homeostasis, are highly relevant to type 2 diabetes etiology, and may be critical to the mechanisms that underlie the rapid positive metabolic effects of bariatric surgery. Very recent studies reveal that several miRNAs are highly enriched in mature EECs and/or in intestinal stem cells that are primed to the EEC lineage. Moreover, functional experiments in enteroids/intestinal organoids suggest that some of these miRNAs may be important for the regulation of EEC differentiation and function. Another report has raised the possibility that EECs secrete miRNAs into circulation. These intriguing findings merit further investigation, particularly as it pertains to EEC miRNAs as novel therapeutic targets in type 2 diabetes and related diseases.

MicroRNAs (miRNAs) are key posttranscriptional regulators of biological pathways that govern lipid metabolic phenotypes. Recent advances in high-throughput small RNA sequencing technology have revealed the complex and dynamic repertoire of miRNAs. Specifically, it has been demonstrated that a single genomic locus can give rise to multiple, functionally distinct miRNA isoforms (isomiR). There are several mechanisms by which isomiRs can be generated, including processing heterogeneity and posttranscriptional modifications, such as RNA editing, exonuclease-mediated nucleotide trimming, and/or nontemplated nucleotide addition (NTA). NTAs are dominant at the 3′-end of a miRNA, are most commonly uridylation or adenlyation events, and are catalyzed by one or more of several nucleotidyl transferase enzymes. 3′ NTAs can affect miRNA stability and/or activity and are physiologically regulated, whereas modifications to the 5′-ends of miRNAs likely alter miRNA targeting activity. Recent evidence also suggests that the biogenesis of specific miRNAs, or small RNAs that act as miRNAs, can occur through unconventional mechanisms that circumvent key canonical miRNA processing steps. The unveiling of miRNA diversity has significantly added to our view of the complexity of miRNA function. In this review we present the current understanding of the biological relevance of isomiRs and their potential role in regulating lipid metabolism.

In recent years, microRNAs (miRNA) have emerged as important posttranscriptional regulators of gene expression in a wide variety of biological pathways. Since the discovery of the liver-specific miRNA-122 (miR-122) and its critical role in hepatic function, numerous additional miRNAs have been implicated in lipid metabolism. It is now apparent that lipid homeostasis is governed in part by an intricate web of miRNA activity. miRNAs are thought to confer robustness against environmental changes, such as diet modifications. Therefore, naturally occurring genetic variation that perturbs miRNA expression and/or function is likely to contribute to interindividual variability in lipid phenotypes. Although the field is still in its infancy, this review describes the growing evidence for miRNA-related genetic variation as etiological factors in lipid disorders. Specific examples, including a variant in a miRNA transcriptional control element that leads to dyslipidemia as well as a variant in a miRNA target site that modulates the effect of diet on plasma lipid levels, are discussed. Finally, the utility of recent systems genetics approaches to uncover hidden miRNA-related genetic associations with lipid disorders are considered, thereby illuminating the needles in the genetic haystack.

Nano size Polypyrrole (PPY) was synthesized electrochemically in presence of three different surfactants namely CTAB, CSA and TTX-100. Cyclic voltammetric studies showed an irreversible oxidation of pyrrole in the presence of surfactants. The currents of the broad peak were 4.26mA, 5.3mA and 8.6mA for PPY/CTAB, PPY/CSA and PPY/TTY-100 respectively. The FT IR spectra of electrochemically synthesized PPY in presence and absence of surfactants were analyzed and the prominent peaks were assigned properly. The presence of N-H group in both cases suggests the presence of pyrrole units in the polymer. From the XRD results the particle sizes of all the four PPYs were determined and the values are as follows: PPY (34 nm), PPY/CSA (31 nm), PPY/CTAB (30nm), PPY/TTX-100 (29 nm). The PPY/TTX-100 exhibited highest electrical conductivity (2.04 ×10−3 S/cm) among other PPY synthesized (PPY/CTAB - 1.13 ×10−3 S/cm, PPY/CSA - 1.09×10−3 S/cm and PPY - 1.27 ×10−3 S/cm). SEM images of PPY/CSA showed agglomerated small polymeric fragments whereas PPY/CTAB showed large sized polymeric plates. Interestingly PPY/TTX–100 showed uniform hollow structure. The presence of surfactants has resulted in uniform polymer formation.

Cellular gene expression is governed by a complex, multi-faceted network of regulatory interactions. In the last decade, microRNAs (miRNAs) have emerged as critical components of this network. miRNAs are small, non-coding RNA molecules that serve as post-transcriptional regulators of gene expression. Although there has been substantive progress in our understanding of miRNA-mediated gene regulation, the mechanisms that control the expression of the miRNAs themselves are less well understood. Identifying the factors that control miRNA expression will be critical for further characterizing miRNA function in normal physiology and pathobiology. We describe recent progress in the efforts to map genomic regions that control miRNA transcription (such as promoters). In particular, we highlight the utility of large-scale “-omic” data, such as those made available by the ENCODE and the NIH Roadmap Epigenomics consortiums, for the discovery of transcriptional control elements that govern miRNA expression. Finally, we discuss how integrative analysis of complementary genetic datasets, such as the NHGRI Genome Wide Association Studies Catalog, can predict novel roles for transcriptional mis-regulation of miRNAs in complex disease etiology.

Uterine leiomyoma is a common benign uterine tumor of women in the reproductive age group. Although the common symptoms of leiomyoma are menorrhagia and dysmenorrhoea, this patient presenting as a near collapse is a rare finding. The patient presented with abdominal pain and worsening anemia within a span of hours and hypotension, tachycardia, and tachypnoea needing urgent surgical intervention and blood transfusion and intensive care support is relatively rare. Though every effort to know the exact cause of intraperitoneal hemorrhage in this patient was taken, the rarer diagnosis of capsular venous rupture was not identified prior to surgical intervention. Initially, laparoscopy was introduced first to identify the cause of massive hemorrhage; the approach was changed to open myomectomy keeping in mind the general condition of the patient. Hence, for any patient with a prior diagnosis of myoma with hemodynamic instability, the rarer diagnosis of leiomyomatous capsular venous erosion should be a differential diagnosis to aid in the appropriate management of the women. The team of interdisciplinary expertise will definitely improve the outcomes in such cases.

MicroRNAs (miRNAs) are approximately 22-nt RNA segments that are involved in the regulation of protein expression primarily by binding to one or more target sites on an mRNA transcript and inhibiting translation. MicroRNAs are likely to factor into multiple developmental pathways, multiple mechanisms of gene regulation, and underlie an array of inherited disease processes and phenotypic determinants. Several computational programs exist to predict miRNA targets in mammals, fruit flies, worms, and plants. However, to date, there is no systematic collection and description of miRNA targets with experimental support. We describe a database, TarBase, which houses a manually curated collection of experimentally tested miRNA targets, in human/mouse, fruit fly, worm, and zebrafish, distinguishing between those that tested positive and those that tested negative. Each positive target site is described by the miRNA that binds it, the gene in which it occurs, the nature of the experiments that were conducted to test it, the sufficiency of the site to induce translational repression and/or cleavage, and the paper from which all these data were extracted. Additionally, the database is functionally linked to several other useful databases such as Gene Ontology (GO) and UCSC Genome Browser. TarBase reveals significantly more experimentally supported targets than even recent reviews claim, thereby providing a comprehensive data set from which to assess features of miRNA targeting that will be useful for the next generation of target prediction programs. TarBase can be accessed at http://www.diana.pcbi.upenn.edu/tarbase.

Type 2 inflammation is associated with epithelial cell responses, including goblet cell hyperplasia, that promote worm expulsion during intestinal helminth infection. How these epithelial responses are regulated remains incompletely understood. Here, we show that mice deficient in the prostaglandin D2 (PGD2) receptor CRTH2 and mice with CRTH2 deficiency only in nonhematopoietic cells exhibited enhanced worm clearance and intestinal goblet cell hyperplasia following infection with the helminth Nippostrongylus brasiliensis. Small intestinal stem, goblet, and tuft cells expressed CRTH2. CRTH2-deficient small intestinal organoids showed enhanced budding and terminal differentiation to the goblet cell lineage. During helminth infection or in organoids, PGD2 and CRTH2 down-regulated intestinal epithelial Il13ra1 expression and reversed Type 2 cytokine–mediated suppression of epithelial cell proliferation and promotion of goblet cell accumulation. These data show that the PGD2–CRTH2 pathway negatively regulates the Type 2 cytokine–driven epithelial program, revealing a mechanism that can temper the highly inflammatory effects of the anti-helminth response.

Genetic approaches in model organisms have consistently demonstrated that molecular traits such as gene expression are under genetic regulation, similar to clinical traits. The resulting expression quantitative trait loci (eQTL) have revolutionized our understanding of genetic regulation and identified numerous candidate genes for clinically relevant traits. More recently, these analyses have been extended to other molecular traits such as protein abundance, metabolite levels, and miRNA expression. Here, we performed global hepatic eQTL and microRNA expression quantitative trait loci (mirQTL) analysis in a population of Diversity Outbred mice fed two different diets. We identified several key features of eQTL and mirQTL, namely differences in the mode of genetic regulation (cisor trans) between mRNA and miRNA. Approximately 50% of mirQTL are regulated by a trans-acting factor, compared to ∼25% of eQTL. We note differences in the heritability of mRNA and miRNA expression and variance explained by each eQTL or mirQTL. In general, cis-acting variants affecting mRNA or miRNA expression explain more phenotypic variance than trans-acting variants. Finally, we investigated the effect of diet on the genetic architecture of eQTL and mirQTL, highlighting the critical effects of environment on both eQTL and mirQTL. Overall, these data underscore the complex genetic regulation of two well-characterized RNA classes (mRNA and miRNA) that have critical roles in the regulation of clinical traits and disease susceptibility

Although embryonic brain development and neurodegeneration have received considerable attention, the events that govern postnatal brain maturation are less understood. Here, we identify the miR-29 family to be strikingly induced during the late stages of brain maturation. Brain maturation is associated with a transient, postnatal period of de novonon-CG (CH) DNA methylation mediated by DNMT3A. We examine whether an important function of miR-29 during brain maturation is to restrict the period of CH methylation via its targeting of Dnmt3a. Deletion of miR-29 in the brain, or knockin mutations preventing miR-29 to specifically target Dnmt3a, result in increased DNMT3A expression, higher CH methylation, and repression of genes associated with neuronal activity and neuropsychiatric disorders. These mouse models also develop neurological deficits and premature lethality. Our results identify an essential role for miR-29 in restricting CH methylation in the brain and illustrate the importance of CH methylation regulation for normal brain maturation.

Brake friction linings are made of materials with a highly complex formulation that helps in improving the braking performance. The selection of friction materials with good physical, mechanical, and thermal properties is vital, which will decide the braking performance. Apart from giving good physio-mechanical properties, metallic fillers act as heat dissipaters. The objective of this work is to study the synergetic effect of prominent heat dissipaters, namely copper fibers, brass fibers, and zinc powders. Three simplified formulations were developed with 10, 14, and 18 wt.% of these heat dissipaters and named DB1, DB2, and DB3, respectively. It was observed that the addition of heat dissipaters increased the thermal properties. Tribological properties are tested based on SAE J661 standards. It was observed that DB2 had a consistent and higher coefficient of friction of 0.503 with a higher wear rate (7.6%) while DB3 had adequate μand lower wear rate. The same batches of brake pads were tested in an inertia brake dynamometer following JASO C406 and a wear test was carried out. It was observed that % fade and % recovery were better for DB2 in both cycles. The wear rate in terms of thickness was lesser for DB2 followed by DB1 and DB3. The wear mechanism was analyzed using a scanning electron microscope. The preference selection index method of optimization was used to evaluate the overall performance parameters of the brake friction composites. Heat dissipaters with 14 wt.% have proved to be the better performers, followed by 10 and 18 wt.%.

Fibrolamellar carcinoma (FLC) is a rare, therapeutically intractable liver cancer that disproportionately affects youth. Although FLC tumors exhibit a distinct gene expression profile, the chromatin regulatory landscape and the genes most critical for tumor cell survival remain unclear. Here, we use chromatin run-on sequencing to discover ∼7,000 enhancers and 141 enhancer hotspots activated in FLC relative to nonmalignant liver. Bioinformatic analyses reveal aberrant ERK/MEK signaling and candidate master transcriptional regulators. We also define the genes most strongly associated with hotspots of FLC enhancer activity, including CA12and SLC16A14. Treatment of FLC cell models with inhibitors of CA12 or SLC16A14 independently reduce cell viability and/or significantly enhance the effect of the MEK inhibitor cobimetinib. These findings highlight molecular targets for drug development, as well as drug combination approaches.