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Background: Chinese hamster ovarian (CHO) cells are widely employed in biotechnology for the production of recombinant proteins. Extending the life span of CHO cells and inhibiting the loss of producing cell population through the inhibition of apoptosis can benefit the productivity of those cells. In this study, we aimed to screen and evaluate the impact of some caspase-7 inhibitor candidates on the lifespan of CHO cells. Results: Through virtual screening and molecular docking, risperidone was screened and selected as a potential inhibitor of caspase-7 in CHO cells. The results of MTT assay revealed that the cytotoxicity of risperidone at all concentrations was lower than 50%, and thus it can be suggested as a safe treatment for CHO cells. Annexin V apoptosis and flow cytometry assays revealed that risperidone at 1, 25, and 50 µM concentrations inhibited apoptosis 72 h post-treatment through caspase-7 inhibition. Although gene expression analysis through qRT-PCR demonstrates that risperidone did not affect caspase-7 gene expression. Conclusions: This bioinformatics and experimental study suggests risperidone as a caspase-7 inhibitor with the potential to extend the lifespan of CHO cells and offers possible opportunities in biotechnology.How to cite: Kafi S, Najafi S, Mahnam K, et al. Virtual screening and experimental analysis of caspase-7 inhibitors as candidates for extending the lifespan of CHO cells. Electron J Biotechnol 2024;71. https://doi.org/10.1016/j.ejbt.2024.04.007.

The potential of L-glutathione (GSH) (0, 4, 16, 32 and 64 mM) to improve the post-harvest quality and antioxidant capacity of strawberries was investigated during storage (0, 5, 10, and 15 days) in this study. Results showed that weight loss in fruits treated with 64 mM GSH was significantly lower than the control. GSH treatments resulted in higher levels of total phenol content and antioxidant capacity in treated fruits of strawberry. Based on the results, GSH 64 mM significantly increased the levels of total flavonoid, anthocyanin, ascorbic acid, total soluble protein, antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), guaiacol peroxidase (POD), ascorbate peroxidase (APX) and Phenylalanine ammonia-lyase (PAL). In addition, GSH 64 mM decreased Malondialdehyde (MDA) levels and prevented cell membrane lipid peroxidation. In conclusion, the results of the present study showed that the use of GSH 64 mM may be a promising strategy to improve the marketability, quality and antioxidant capacity of strawberry fruits during storage.

One of the multifactorial worldwide health syndromes is diabetes mellitus which is increasing at a disturbing rate. The inhibition of α-glucosidase, an enzyme that catalyzes starch hydrolysis in the intestine, is one helpful therapeutic approach for controlling hyperglycemia related to type-2 diabetes. To discover α-glucosidase inhibitors, some 2-hydrazolyl-4-thiazolidinone hybrids (3a-e) were synthesized from new one-pot reaction procedures. Next, their chemical structures were confirmed by 1H NMR, 13C NMR, and FT-IR spectra, and elemental analysis technique. Then, the α-glucosidase inhibitory activity of the titled compounds was evaluated. Among them, derivatives 3b and 3c revealed the highest activity against α-glucosidase compared to acarbose as a drug. Enzyme kinetic studies of the most active derivative (3b) indicated a competitive inhibition. Finally, molecular modeling studies were accomplished to describe vital interactions of the most potent compounds (3b and 3c) with the α-glucosidase enzyme.

Compressions are prevalent in industrial applications and are notable for their substantial energy consumption. Therefore, the simulation and analysis of the compression process are essential for maintenance and energy conservation efforts. These systems are prone to potentially unstable surge conditions, necessitating the use of traditional anti-surge valves that result in considerable energy losses. Ensuring the near-optimal operation of these systems is critical to minimizing energy consumption.In this article, a conceptual framework for a cylinder-piston mechanism is delineated, intended for design and operation as an active surge control system. Additionally, a modular quasi-one-dimensional model is articulated for the transient simulation of an industrial compression system, which integrates models for both the anti-surge and active control systems. The manuscript presents a novel design, featured by a cylinder-piston system integrated with a robust controller, posited as a potential alternative to traditional anti-surge systems. The effectiveness of this design in expanding the operational envelope of the compression system and surge prevention is rigorously examined. Moreover, a thermodynamic model, grounded in the fundamental laws of mass, momentum, and energy conservation, is applied to each component of the system. Furthermore, the manuscript explores the benefits of the innovative design in achieving a marked decrease in energy wastage. Simulation results from a test scenario reveals that the implementation of the cylinder-piston design, as opposed to the conventional anti-surge system, can diminish energy losses and associated pollutant emissions by approximately 33 percent.

In this study, high-Fidelity Large eddy simulation (LES) data was used to create a surrogate turbulence model to enhance the estimation of turbulent heat flux in a Reynolds averaged Navier Stokes (RANS) solver for film cooling applications. The LES simulation (A flat plate film cooling application with a Reynolds number of 17,382, a blowing ratio of 1 and a density ratio of 2) was validated by comparison with experimental data. A correlation analysis was performed to identify the most influential parameters, and temperature gradient, velocity gradient, turbulent dissipation rate, shear strain rate, and turbulent viscosity ratio were selected as the most effective parameters. A second model was developed to create a Galilean-invariant model that remains unaffected by changes in the coordinate system. Neural network was applied to the LES training data to propose a modified surrogate dynamic turbulent Prandtl number to be applied to the k − ε realizable RANS simulation. The first surrogate model was implemented to two different geometries, one geometry was a flat plate film cooling similar to the training data but with different flow conditions, and another one was a more complex geometry with pressure gradient. The second model was only implemented on the second geometry and produced similar results. The surrogate models predicted a more accurate thermal field near the cooling wall (up-to 58 % error reduction) with a computational time complexity similar to the base RANS solver.

Colorectal cancer (CRC) is a multi-stage process initiated through the formation of a benign adenoma, progressing to an invasive carcinoma and finally metastatic spread. Tumour cells must adapt their metabolism to support the energetic and biosynthetic demands associated with disease progression. As such, targeting cancer cell metabolism is a promising therapeutic avenue in CRC. However, to identify tractable nodes of metabolic vulnerability specific to CRC stage, we must understand how metabolism changes during CRC development. Here, we use a unique model system – comprising human early adenoma to late adenocarcinoma. We show that adenoma cells transition to elevated glycolysis at the early stages of tumour progression but maintain oxidative metabolism. Progressed adenocarcinoma cells rely more on glutamine-derived carbon to fuel the TCA cycle, whereas glycolysis and TCA cycle activity remain tightly coupled in early adenoma cells. Adenocarcinoma cells are more flexible with respect to fuel source, enabling them to proliferate in nutrient-poor environments. Despite this plasticity, we identify asparagine (ASN) synthesis as a node of metabolic vulnerability in late-stage adenocarcinoma cells. We show that loss of asparagine synthetase (ASNS) blocks their proliferation, whereas early adenoma cells are largely resistant to ASN deprivation. Mechanistically, we show that late-stage adenocarcinoma cells are dependent on ASNS to support mTORC1 signalling and maximal glycolytic and oxidative capacity. Resistance to ASNS loss in early adenoma cells is likely due to a feedback loop, absent in late-stage cells, allowing them to sense and regulate ASN levels and supplement ASN by autophagy. Together, our study defines metabolic changes during CRC development and highlights ASN synthesis as a targetable metabolic vulnerability in later stage disease.

Summary: Background: The high variability in clinical and metabolic presentations of inborn errors of cobalamin (cbl) metabolism (IECM), such as the cblC/epicblC types with combined deficits in methylmalonyl-coA mutase (MUT) and methionine synthase (MS), are not well understood. They could be explained by the impaired expression/activity of enzymes from other metabolic pathways. Methods: We performed metabolomic, genomic, proteomic, and post-translational modification (PTM) analyses in fibroblasts from three cblC cases and one epi-cblC case compared with three cblG cases with specific MS deficits and control fibroblasts. Findings: CblC patients had metabolic profilings consistent with altered urea cycle, glycine, and energy mitochondrial metabolism. Metabolomic analysis showed partial disruption and increased glutamate/ketoglutarate anaplerotic pathway of the tricarboxylic acid cycle (TCA), in patient fibroblasts. RNA-seq analysis showed decreased expression of MT-TT (mitochondrial tRNA threonine), MT-TP (mitochondrial tRNA proline), OXCT1 (succinyl CoA:3-oxoacid CoA transferase deficiency), and MT-CO1 (cytochrome C oxidase subunit 1). Proteomic changes were observed for key mitochondrial enzymes, including NADH:ubiquinone oxidoreductase subunit A8 (NDUFA8), carnitine palmitoyltransferase 2 (CPT2), and ubiquinol-cytochrome C reductase, complex III subunit X (UQCR10). Propionaldehyde addition in ornithine aminotransferase was the predominant PTM in cblC cells and could be related with the dramatic cellular increase in propionate and methylglyoxalate. It is consistent with the decreased concentration of ornithine reported in 3 cblC cases. Whether the changes detected after multi-omic analyses underlies clinical features in cblC and cblG types of IECM, such as peripheral and central neuropathy, cardiomyopathy, pulmonary hypertension, development delay, remains to be investigated. Interpretation: The omics-related effects of IECM on other enzymes and metabolic pathways are consistent with the diversity and variability of their age-related metabolic and clinical manifestations. PTMs are expected to produce cumulative effects, which could explain the influence of age on neurological manifestations. Funding: French Agence Nationale de la Recherche (Projects PREDICTS and EpiGONE) and Inserm.

Introduction: Intramedullary nailing has become the gold standard in surgical treatment for long bones shaft fractures. Even in expert hands, many intraoperative complications may occur. Clinical case: We report an unusual complication of trapped bone fragments between the reamer and the cortex during intramedullary nailing of a simple femur shaft fracture. All Attempts to withdraw the reamer were unsuccessful. The fragment had to be removed through an open incision.This complication was probably caused by a migrated chip of cortical bone fragment from the inner cortex which caused incarceration of the reamer. Conclusion: If a loose intramedullary fragment is noted during reaming or nailing, it must absolutely not be ignored.

Performance of thermoacoustic (TA) systems are very complex, and their performance optimization is a difficult task, and most TA systems have relatively very low efficiencies. The main objective of this study was to develop an exergy analysis to introduce a guide for a designer, how to optimize the performance of a TA system. We have applied this methodology to a coupled thermoacoustic engine and refrigerator with a standing wave in the engine and a traveling wave in the refrigerator. In an optimization procedure, a lumped systems numerical analysis was performed to predict the main parameters of the thermoacoustic system. The numerical method was validated by comparison with numerical and experimental results in the literature. The cost function was the total system coefficient of performance, and design parameters include the refrigerator regenerator length, location, and hydraulic radius. The performance index was decomposed into three main subsystem indices, i.e., the efficiency of the engine, the efficiency of the resonator, and the coefficient of performance of the refrigerator. The optimum point for each component was found, and the total system optimal point which is a result of thermoacoustic interaction of all segments were analyzed. It was illustrated how a change in design parameters could affect the pressure and velocity amplitude and phase distribution in different subsystems, which help the designer deal with the challenge of the optimal parameter selection to enhance the system performance. Finally, an exergy analysis was introduced to assess loss mechanism's contribution to each refrigerator's component. This result in a roadmap for optimization of the system. A new parameter called non-dimensional exergy loss portion (NELP) was introduced, which may significantly show the improvement potential in each system component.

Introduction: Subtalar dislocation is defined by a simultaneous total separation of the talocalcaneal and talonavicular joints. Lateral variety is the rarest. It is the result of a high energy trauma. It occurs following forced eversion, with the foot locked in a rut with pressure on the lateral aspect of the leg. Articular fractures of the calcaneus occur as a result of axial impact on the greater tuberosity.We report the case of a lateral subtalar dislocation associated with a calcaneal fracture. Case presentation: A 62-year-old man who presented to the emergency room following a 9 m fall. Initial examination revealed a swelling of the right foot and ankle with a deformity giving the appearance of a “flat foot” without skin opening. The neuro-vascular examination was normal.A radiological assessment including an antero-posterior and lateral X-ray of the right ankle and a axial view if the calcaneus, showed a calcaneal articular fracture and a lateral dislocation of the subtalar joint. CT scan confirmed the diagnosis. Closed reduction by external maneuvers failed. Through a lateral mini-approach centered on the subtalar joint, open reduction of the dislocation was achieved and controlled fluoroscopically. Osteosynthesis of the calcaneal fracture was performed using a percutaneous pinning. Consolidation was obtained at 3 months post-operatively.At the last follow-up, the patient reported occasional pain. The radiological assessment showed early arthritic changes of the subtalar joint. Conclusion: In rare cases of subtalar dislocation, the direction of displacement, associated bone injuries, and open reduction would be responsible for a poor functional outcome. At present, there is no certain way to avoid subtalar osteoarthritis.

Compression systems are extensively used in the petrochemical industry. They are very intensive energy consumers, and simulation and analysis of this process are vital for their better maintenance and limiting their energy consumption. Here we follow an industrial case study to model and simulate a compression loop of the petrochemical industry and show potentials for energy savings and environmental improvements In this paper we introduce a modular quasi-one-dimensional model to simulate a two-step seven-stage compressor and all components of the compression loop, including the surge control system. For each component, including pipes, separation drums, heat exchangers, and compressors, a thermodynamic model based on the mass, momentum, and energy conservation laws is obtained, then each component of the system can be presented as an independent module. By stacking all modules together, the entire compression system is modeled. Finally, a numerical simulation is performed for many different scenarios to study the behavior of the surge phenomenon, the surge control system, and especially the effects of the intentional anti-surge valve openness for extra surge margin, which results in huge energy loss and environmental impacts. This simulation shows that removing the intentional openness of the anti-surge valves will reduce about 15.8% of the CO2 emissions, and the power consumption required to run the turbine may be reduced up to 1.8 MW.

Abstract: Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare and aggressive leukemia for which we developed a nationwide network to collect data from new cases diagnosed in France. In a retrospective, observational study of 86 patients (2000-2013), we described clinical and biological data focusing on morphologies and immunophenotype. We found expression of markers associated with plasmacytoid dendritic cell origin (HLA-DRhigh, CD303+, CD304+, and cTCL1+) plus CD4 and CD56 and frequent expression of isolated markers from the myeloid, B-, and T-lymphoid lineages, whereas specific markers (myeloperoxidase, CD14, cCD3, CD19, and cCD22) were not expressed. Fifty-one percent of cytogenetic abnormalities impact chromosomes 13, 12, 9, and 15. Myelemia was associated with an adverse prognosis. We categorized chemotherapeutic regimens into 5 groups: acute myeloid leukemia (AML)–like, acute lymphoid leukemia (ALL)–like, lymphoma (cyclophosphamide, doxorubicin, vincristine, and prednisone [CHOP])–like, high-dose methotrexate with asparaginase (Aspa-MTX) chemotherapies, and not otherwise specified (NOS) treatments. Thirty patients received allogeneic hematopoietic cell transplantation (allo-HCT), and 4 patients received autologous hematopoietic cell transplantation. There was no difference in survival between patients receiving AML-like, ALL-like, or Aspa-MTX regimens; survival was longer in patients who received AML-like, ALL-like, or Aspa-MTX regimens than in those who received CHOP-like regimens or NOS. Eleven patients are in persistent complete remission after allo-HCT with a median survival of 49 months vs 8 for other patients. Our series confirms a high response rate with a lower toxicity profile with the Aspa-MTX regimen, offering the best chance of access to hematopoietic cell transplantation and a possible cure.

Placer mining is a mineral extraction method in floodplains that involves the removal of earth material to access mineral-laden sediments, a process that can have significant and long-term impacts on aquatic ecosystems. Given the widespread nature of mining, new tools are required to monitor the potential watershed-scale ecological impacts of placer mining. This study adapted and evaluated a deep learning model – a U-Net convolution neural network, and compared it to a traditional image classification method – random forests (RF) – to detect and quantify the area of post-placer mining disturbance at the watershed scale. Overall, both random forest and U-Net models performed well at classifying digitized image samples where placer disturbances were mapped. Sensitivity in placer classification was high, with both modelling frameworks achieving at least 75% accuracy in the classification of digitized placer samples in 7 out of 12 modelling scenarios. Misclassification of non-placer pixels as placer was highly variable among different models, data configurations, study sites, and time periods. Commission errors (i.e., incorrectly classifying a non-placer pixel as placer) were typically the result of models labelling water areas or forest areas as placer – errors which may have only marginal practical significance. In general, U-Net models performed better in terms of minimizing misclassification errors, whereas RF models performed slightly better in classifying known placer pixels. We conclude with discussions on the advantages of deploying U-Net and RF models for placer detection, challenges that may be encountered in operational systems that employ the models, and identifying outstanding issues which need to be addressed in future placer modelling studies.

Abstract: Chronic lymphocytic leukemia (CLL), the most common adulthood leukemia in Western countries, is a very heterogeneous disease characterized by a peripheral accumulation of abnormal CD5+ B lymphocytes in the immune system. Despite new therapeutic developments, there remains an unmet medical need for CLL. Here, we demonstrate that the use of N-methylated thrombospondin-1 (TSP-1)–derived peptides is an efficient way to kill the malignant CLL cells, including those from high-risk individuals with poor clinical prognosis, del11q, del17p, 2p gain, or complex karyotype. PKT16, our hit N-methylated peptide, triggers the elimination of the leukemic cells, sparing the nontumor cells, including the hematopoietic precursors, and reduces the in vivo tumor burden of a CLL-xenograft mice model. A complementary analysis underscores the improved cytotoxic efficiency of PKT16 compared with the previously described TSP-1–derived probes, such as PKHB1. PKT16 elicits an original caspase-independent programmed necrotic mode of cell death, different from necroptosis or ferroptosis, implicating an intracellular Ca2+ deregulation that provokes mitochondrial damage, cell cycle arrest, and the specific death of the malignant CLL cells. The activation of the Gαi proteins and the subsequent drop of cyclic adenosine monophosphate levels and protein kinase A activity regulate this cytotoxic cascade. Remarkably, PKT16 induces the molecular hallmarks of immunogenic cell death, as defined by the calreticulin plasma membrane exposure and the release of adenosine triphosphate and high-mobility group box 1 protein from the dying CLL cells. Thus, PKT16 appears to be able to stimulate an anticancer in vivo immune response. Collectively, our results pave the way toward the development of an efficient strategy against CLL.

Neuroblastoma is one of the commonest and deadliest solid tumours of childhood, and is thought to result from disrupted differentiation of the developing sympathoadrenergic lineage of the neural crest. Neuroblastoma exhibits intra- and intertumoural heterogeneity, with high risk tumours characterised by poor differentiation, which can be attributable to MYCN-mediated repression of genes involved in neuronal differentiation. MYCN is known to co-operate with oncogenic signalling pathways such as Alk, Akt and MEK/ERK signalling, and, together with c-MYC has been shown to be activated by Wnt signalling in various tissues. However, our previous work demonstrated that Wnt3a/Rspo2 treatment of some neuroblastoma cell lines can, paradoxically, decrease c-MYC and MYCN proteins. This prompted us to define the neuroblastoma-specific Wnt3a/Rspo2-driven transcriptome using RNA sequencing, and characterise the accompanying changes in cell biology. Here we report the identification of ninety Wnt target genes, and show that Wnt signalling is upstream of numerous transcription factors and signalling pathways in neuroblastoma. Using live-cell imaging, we show that Wnt signalling can drive differentiation of SK-N-BE(2)-C and SH-SY5Y cell-lines, but, conversely, proliferation of SK-N-AS cells. We show that cell-lines that differentiate show induction of pro-differentiation BMP4 and EPAS1 proteins, which is not apparent in the SK-N-AS cells. In contrast, SK-N-AS cells show increased CCND1, phosphorylated RB and E2F1 in response to Wnt3a/Rspo2, consistent with their proliferative response, and these proteins are not increased in differentiating lines. By meta-analysis of the expression of our 90 genes in primary tumour gene expression databases, we demonstrate discrete expression patterns of our Wnt genes in patient cohorts with different prognosis. Furthermore our analysis reveals interconnectivity within subsets of our Wnt genes, with one subset comprised of novel putative drivers of neuronal differentiation repressed by MYCN. Assessment of β-catenin immunohistochemistry shows high levels of β-catenin in tumours with better differentiation, further supporting a role for canonical Wnt signalling in neuroblastoma differentiation.

The mosquito-borne flaviviruses include important human pathogens such as dengue, Zika, West Nile, and yellow fever viruses, which pose a serious threat for global health. Recent genetic screens identified endoplasmic reticulum (ER)-membrane multiprotein complexes, including the oligosaccharyltransferase (OST) complex, as critical flavivirus host factors. Here, we show that a chemical modulator of the OST complex termed NGI-1 has promising antiviral activity against flavivirus infections. We demonstrate that NGI-1 blocks viral RNA replication and that antiviral activity does not depend on inhibition of the N-glycosylation function of the OST. Viral mutants adapted to replicate in cells deficient of the OST complex showed resistance to NGI-1 treatment, reinforcing the on-target activity of NGI-1. Lastly, we show that NGI-1 also has strong antiviral activity in primary and disease-relevant cell types. This study provides an example for advancing from the identification of genetic determinants of infection to a host-directed antiviral compound with broad activity against flaviviruses.

Wilms tumors (WTs) have a complex etiology, displaying genetic and epigenetic changes, including loss of imprinting (LOI) and tumor suppressor gene silencing. To identify new regions of epigenetic perturbation in WTs, we screened kidney and tumor DNA using CpG island (CGI) tags associated with cancer-specific DNA methylation changes. One such tag corresponded to a paralog of the glioma pathogenesis-related 1/related to testis-specific, vespid, and pathogenesis proteins 1 (GLIPR1/RTVP-1) gene, previously reported to be a tumor-suppressor gene silenced by hypermethylation in prostate cancer. Here we report methylation analysis of the GLIPR1/RTVP-1 gene in WTs and normal fetal and pediatric kidneys. Hypomethylation of the GLIPR1/RTVP-1 5'-region in WTs relative to normal tissue is observed in 21/24 (87.5%) of WTs analyzed. Quantitative analysis of GLIPR1/RTVP-1 expression in 24 WTs showed elevated transcript levels in 16/24 WTs (67%), with 12 WTs displaying in excess of 20-fold overexpression relative to fetal kidney (FK) control samples. Immunohistochemical analysis of FK and WT corroborates the RNA expression data and reveals high GLIPR1/RTVP-1 in WT blastemal cells together with variable levels in stromal and epithelial components. Hypomethylation is also evident in the WT precursor lesions and nephrogenic rests (NRs), supporting a role for GLIPR1/RTVP-1 deregulation early in Wilms tumorigenesis. Our data show that, in addition to gene dosage changes arising from LOI and hypermethylation-induced gene silencing, gene activation resulting from hypomethylation is also prevalent in WTs.

A novel third-generation biorefinery approach, including two paths of Ethanol/methane production pathway (EMP) and the direct methane production pathway (DMP), for astaxanthin and ethanol and biogas production from the freshwater microalgae Haematococcus pluvialis was developed previously. To ensure its environmental sustainability, a comprehensive life cycle assessment (LCA) study was conducted based on 1-GJ energy generation from biomethane as the functional unit. Results indicate that the EMP pathway had higher environmental impacts on all categories due to more stages and chemicals/energy consumption (at least five times greater effect). Results showed that while the enzymatic hydrolysis step followed by the fermentation stage was the main contributor to all environmental categories in the EMP route, astaxanthin induction dominated all environmental categories in the DMP route. The results showed that sodium nitrate, phosphate salts, inoculum sludge, acetone, and electricity had considerable environmental impacts. Moreover, despite low enzyme usage in enzymatic hydrolysis, these proteins significantly impacted all environmental categories in this stage. The baseline analysis concluded that to produce 1 GJ energy from methane, about 88 kg and 13 kg CO 2 were generated from the EMP and DMP pathways, respectively. A sensitivity analysis was also conducted to compare various ratios of chemicals, such as phosphate salts, with high contributions to enzymatic hydrolysis and astaxanthin induction stages in the EMP and DMP routes, respectively. Finally, the LCA results revealed that the DMP pathway is more environmentally friendly with the same economic value of biomethane and astaxanthin production. This LCA study updated the data related to the environmental assessment of processes to utilize H. pluvialis to produce biofuels and astaxanthin simultaneously., Competing Interests: Declaration of competing interest The current investigation is supported by the University of Nevada, Reno. Any opinions, findings, conclusions, or recommendations expressed in this paper are those of the authors and do not necessarily reflect the views of the University of Nevada, Reno., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The pandemic prevalence of COVID-19 has become a very serious global health issue. Scientists all over the world have been seriously attempting in the discovery of a drug to combat SARS-CoV-2. It has been found that RNA-dependent RNA polymerase (RdRp) plays a crucial role in SARS-CoV-2 replication, and thus could be a potential drug target. Here, comprehensive computational approaches including drug repurposing and molecular docking were employed to predict an effective drug candidate targeting RdRp of SARS-CoV-2. This study revealed that Rifabutin, Rifapentine, Fidaxomicin, 7-methyl-guanosine-5'-triphosphate-5'-guanosine and Ivermectin have a potential inhibitory interaction with RdRp of SARS-CoV-2 and could be effective drugs for COVID-19. In addition, virtual screening of the compounds from ZINC database also allowed the prediction of two compounds (ZINC09128258 and ZINC09883305) with pharmacophore features that interact effectively with RdRp of SARS-CoV-2, indicating their potentiality as effective inhibitors of the enzyme. Furthermore, ADME analysis along with analysis of toxicity was also undertaken to check the pharmacokinetics and drug-likeness properties of the two compounds. Comparative structural analysis of protein-inhibitor complexes revealed that the amino acids Y32, K47, Y122, Y129, H133, N138, D140, T141, S709 and N781 are crucial for drug surface hotspot in the RdRp of SARS-CoV-2., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Introduction: Quadrilateral plate fractures constitute one of the most challenging components of acetabular fractures. The objective of this study is to describe and evaluate the novel technique of buttress screw fixation of the quadrilateral plate component of the acetabular fractures., Patients & Methods: Forty cases of acetabular fracture with associated quadrilateral plate component were included in the study. Mean age was 35 years (range, 16-68years), with a mean follow-up 16.4 months (range, 9-36months). Fixation of the quadrilateral plate was achieved by one or more buttress screws. The screws were inserted through the reconstruction plate, and placed close to the edge of the pelvic brim. To effectively achieve the 3-point fixation principle, the screw was inserted through the plate hole then outside the bone rubbing on the pelvic surface of the quadrilateral plate., Results: Anatomical reduction of the quadrilateral plate component of the fractures was achieved in all but one patient. The modified Merle D'Aubigné and Postel score was excellent in 13 cases, good in 23 cases, fair in three cases, and poor in one case. No screw displacement or failures were observed during follow-up evaluation. No major complications related to this technique were observed in this series., Conclusion: Buttress screw fixation of the quadrilateral plate fracture component in associated acetabular fractures is a safe and effective technique for reduction and fixation of these challenging fractures with no major complications related to this novel technique., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Differentiated trophoblast cell lineages arise from trophoblast stem (TS) cells. To date such a stem cell population has only been established in the mouse. The objective of this investigation was to establish TS cell populations from rat blastocysts. Blastocysts were cultured individually on a feeder layer of rat embryonic fibroblasts (REFs) in fibroblast growth factor-4 (FGF4) and heparin supplemented culture medium. Once cell colonies were established REF feeder layers could be replaced with REF conditioned medium. The blastocyst-derived cell lines, in either proliferative or differentiated states, did not express genes indicative of ICM-derived tissues. In the proliferative state the cells expressed established stem cell-associated markers of TS cells. Cells ceased proliferation and differentiated when FGF4, heparin, and REF conditioned medium were removed. Differentiation was characterized by a decline of stem cell-associated marker gene expression, the appearance of large polyploid cells (trophoblast giant cells), and the expression of trophoblast differentiation-associated genes. Collectively, the data indicate that the rat blastocyst-derived cell lines not only possess many features characteristic of mouse TS cells but also possess some distinct properties. These rat TS cell lines represent valuable new in vitro models for analyses of mechanisms controlling TS cell renewal and differentiation., (Copyright © 2010 Elsevier Inc. All rights reserved.)

We conducted an observational cross-sectional study to determine if the prevalence of hematologic and metabolic abnormalities in chronic kidney disease (CKD) varied in different ethnic groups. We used a CKD provincial database where a complete data set at the time of registration was available as well as an estimated glomerular filtration rate (eGFR), which showed using the abbreviated MDRD formula that the patients had CKD of stages 3-5. We included patients with self-reported race of Caucasian, Oriental Asian, or South Asian. Primary outcomes were the prevalence of at least one of the following: anemia, hypocalcemia, hyperphosphatemia, hyperparathyroidism, hypoalbuminemia, and three or more laboratory abnormalities. All definitions were consistent with K/DOQI guidelines. When compared with Caucasians, Oriental Asians and South Asians had a higher prevalence of many of the metabolic abnormalities during most stages of CKD and were more likely to have any abnormality at all levels of eGFR. The prevalence of three or more laboratory abnormalities was higher in Oriental Asians at all stages and in South Asians at some levels of eGFR. These results were unchanged or exaggerated when controlled for age, gender, diabetes, and a primary diagnosis of renal disease. Hence, it appears that South Asians and Oriental Asians have more laboratory abnormalities compared with Caucasians at most levels of eGFR.

Pantothenate kinase (PanK) is thought to catalyze the first rate-limiting step in CoA biosynthesis. The full-length cDNA encoding the human PanK1alpha protein was isolated, and the complete human PANK1 gene structure was determined. Bezafibrate (BF), a hypolipidemic drug and a peroxisome proliferator activator receptor-alpha (PPARalpha) agonist, specifically increased hPANK1alpha mRNA expression in human hepatoblastoma (HepG2) cells as a function of time and dose of the drug, compared with hPANK1beta, hPANK2, and hPANK3, which did not significantly increase. Four putative PPARalpha response elements were identified in the PANKIalpha promoter, and BF stimulated hPANK1alpha promoter activity but did not alter the mRNA half-life. Increased hPANK1alpha mRNA resulted in higher hPanK1 protein, localized in the cytoplasm, and elevated PanK enzyme activity. The enhanced hPANK1alpha gene expression translated into increased activity of the CoA biosynthetic pathway and established a higher steady-state CoA level in HepG2 cells. These data are consistent with a key role for PanK1alpha in the control of cellular CoA content and point to the PPARalpha transcription factor as a major factor governing hepatic CoA levels by specific modulation of PANK1alpha gene expression.

Hypergastrinemia is known to cause hyperplasia of the gastric mucosa, especially in gastric enterochromaffinlike (ECL) cells. In some clinical conditions causing hypergastrinemia, such as long-term gastric-acid inhibition and gastric-mucosa atrophy, hyperplastic ECL cells may develop into gastric carcinoid tumors. A newly developed gastrin-receptor antagonist, S-0509, has been reported to block gastrin-induced stimulation of gastric-acid secretion. We therefore investigated whether S-0509 inhibits the omeprazole- and gastrin-stimulated hyperproliferation of gastric mucosa, especially of ECL cells. Daily administration of omeprazole and gastrin in male Sprague-Dawley rats induced marked hypergastrinemia and increased proliferation of gastric-mucosa cells. The numbers of ECL cells and of ECL cells producing messenger RNA for regenerating gene, a potent growth factor for gastric-mucosa cells, were also augmented by long-term administration of omeprazole and gastrin. Coadministration of S-0509 with omeprazole or gastrin almost completely inhibited the omeprazole- and gastrin-induced changes in gastric mucosa, including mucosal thickening and ECL hyperplasia. S-0509 did not induce gastric-mucosa atrophy, even when administered for as long as 4 weeks. In summary, we have found that a newly developed gastrin receptor antagonist, S-0509, inhibits omeprazole- and gastrin-induced mucosal hyperplasia, especially ECL-cell hyperplasia, in rats.

Lafutidine is a novel histamine H(2)-receptor antagonist with a potent and long-lasting anti-acid secretory effect that has also been found to have a potent gastroprotective effect. We investigated the effect of lafutidine on gastric mucosal injury induced in rats with the use of water-immersion restraint stress (WRS) by examining serum calcitonin gene-related peptide (CGRP) concentrations, which we measured with the use of an enzyme immunometric assay. WRS-induced mucosal erosive injury in the stomach was reduced significantly by both lafutidine and famotidine pretreatment (from 7.79 +/- 2.02 mm(2) to 3.09 +/- 0.74 mm(2) and 4.05 +/- 1.18 mm(2), respectively). A single administration of lafutidine or famotidine did not change the serum CGRP concentration from the control value when these drugs were administered without WRS. Lafutidine pretreatment before WRS caused a significant increase in serum CGRP concentration compared with famotidine (lafutidine, 86.64 +/- 9.52 pg/mL; famotidine, 47.55 +/- 4.35 pg/mL; control, 58.43 +/- 6.07 pg/mL). Our results suggest that lafutidine augments CGRP release from the rat stomach when administered before the induction of WRS.

The concentration of type I interferon receptor (IFN-Rc) in the liver is a crucial factor in determining the efficacy of interferon (IFN) therapy in patients with chronic hepatitis C. Retinoic acids (RAs) can enhance the expression of type I IFN-Rc expression. The aim of this study was to investigate whether RAs increase the anti-hepatitis C virus (HCV) effect of IFN through an increase in IFN-Rc. The hepatocellular carcinoma cell line HuH-7 was treated with 10(-7) mol/L all-trans RA (ATRA) and 9-cis RA (9-CRA). Expression of type I IFN-Rc was investigated at both the mRNA and protein levels with the use of real-time quantitative polymerase chain reaction and flow cytometry, respectively. We investigated the anti-HCV effect, using in vitro HCV transfection, by monitoring the level of HCV RNA in the culture medium. ATRA and 9-CRA enhanced the expression of type I IFN-Rc at both the mRNA and protein levels. After IFN-alpha treatment, the activity of 2,5'-oligoadenylate synthetase was enhanced by RAs, and this enhancement was abolished when blocking antibodies had previously been bound to the surface receptors. IFN treatment decreased the concentration of HCV RNA, and this effect was enhanced by treatment with RAs. Our findings suggest that RAs enhance the anti-HCV replication effect of IFN-alpha through up-regulation of type I IFN-Rc in HuH-7 cells.

The growth of human cancer cells expressing peroxisome proliferator-activated receptor-gamma (PPAR-gamma) has been reported to be inhibited by PPAR-gamma ligands. In esophageal squamous-cell carcinoma cell lines T.T, T.Tn, and EC-GI-10, we detected expression of PPAR-gamma and investigated the effects of PPAR-gamma ligands on these cell lines in vitro with the use of troglitazone, pioglitazone, and 15d-PGJ2. Marked growth inhibition by the PPAR-gamma ligands was observed in all cases. The growth-inhibitory effect was evidenced by a dose-dependent inhibition of deoxyribonucleic acid synthesis and was associated with altered cell-cycle progression manifesting G1 arrest. Cell-cycle arrest in T.Tn cells induced by troglitazone could be correlated with an increased level of cyclin-dependent kinase inhibitor p27(Kip1), p21(Cip1/Waf1), and p18(Ink4c). Moreover, troglitazone treatment increased the expression of interleukin-1 alpha, a multifunctional cytokine implicated in antitumor immunity. These findings suggest that troglitazone and other PPAR-gamma ligands have adjuvant or neoadjuvant therapeutic potentials in esophageal cancer.

This paper presents the results of a new process of soil decontamination termed as couple electric-hydraulic gradient assisted by ion exchange medium (CEHIXM) in which a hydraulic gradient is coupled with electric gradient in a non-conventional arrangement for the purpose of electrolysis and ion transport. A suitable ion exchange medium was used to capture and subsequently recover the cations. In this process, heavy metals no longer precipitated in the treated soil near the pretreatment zone. The cations were transported out of the soil and captured in the ion exchange medium. Several laboratory tests were conducted to examine the removal of Pb, Cd, Zn and Mn from sandy soils. The experimental results demonstrated the removal efficiencies more than 97% for all the four heavy metals at an energy expenditure of 345 kWh/m(3) of soil.

An unusual case of a coronary artery fistula with the right atrium is described. Four months following the placement of a stent in a high grade, complex lesion of the proximal right coronary artery, restenosis, and contrast spill with rapid clearance into the right atrial space was evident on coronary angiogram. The calculated shunt fraction was trivial and hemodynamically insignificant.

Background: Superficial and intramural thrombosis are reproducible histopathological features of the porcine coronary oversized stent injury model. Fibrin and its degradation products are chemotactic for mononuclear leukocytes, and promote the proliferation and migration of vascular smooth muscle cells (VSMC) in vitro. The goal of this study was to quantitate the serial histomorphologic evolution of thrombosis, leukocyte infiltration, VSMC proliferation and collagen accumulation following porcine coronary artery stent placement in a porcine model., Methods: Twenty-four normocholesterolemic swine underwent oversized balloon (3.5-4.0 mm) coronary angioplasty and tantalum metal stent placement. Twenty-six different arterial sites were injured, followed by serial sacrifice at day 4 (n = 6), 8 (n = 6), 14 (n = 6), and 28 (n = 6). Quantitative analysis of the neointima was performed using a high resolution video-microscopy interface and a validated histomorphometric software program. Alpha actin-positive VSMC density (per 10(4) micro(2) neointimal area) and collagen-specific picro-sirius red fluorescence (percent of neointima) were quantitated at sites adjacent to and distant from coronary artery stent placement., Results: The percent of total neointimal area occupied by resolving thrombus material was greater at days 4-8 compared to 14-28 days (59-63% vs. 1-2%; P = 0.001). Mononuclear leukocytes were also significantly increased at days 4 and 8 (92 +/- 1 and 70 +/- 8%) compared to days 14-28 (both 3 +/- 3%; P = 0.001), as a percentage of the total neointimal cellularity. Total neointimal cell density did not change (20 +/- 10, 12 +/- 6, 23 +/- 7 and 20 +/- 3 cells/10(4) micro(2); P-value, NS), despite progressive cross-sectional vascular area stenosis reduction from 7 +/- 3% at day 4 to 72 +/- 14% at day 28 (P = 0.001). Percent neointimal fibrinoid thrombus content and mononuclear leukocyte cellularity were correlated in this model (R = 0.81; P < 0.001). Peri-stent neointimal collagen staining exceeded that at vascular sites distant from porcine coronary stent placement by 14 days (29 +/- 6 vs. 15 +/- 3%), and remained greater at 28 days (35 +/- 11 vs. 16 +/- 12%) (both P < 0.05)., Conclusions: Quantitative serial histomorphometry of porcine coronary vascular stent delivery sites demonstrates early (4-8 day) neointimal mononuclear leukocyte infiltration which is histomorphologically and temporally related to intramural fibrinoid thrombosis. Significant vascular stenosis and collagen deposition occurs by 14-28 days at these vascular injury sites. These data suggest a local interaction between thrombotic and inflammatory elements in porcine coronary neointima following oversized stent injury.

The presence of prominent left atrial v-waves following interventricular septal rupture in acute myocardial infarction have been reported in the past. Hemodynamic profile obtained in one particular case highlighted some of the varying aspects of pressure wave, oxygen saturation, and compliance abnormalities that may also be present in such cases. The presence of large left atrial a-waves rather than v-waves was one of the findings.