bluuebunny/crossref_metadata_2025 huggingface dataset:
Root-Zone CO2 Concentration Affects Partitioning and Assimilation of Carbon in Oriental Melon Seedlings
Authors: Han Xintong , Jing Yuna , Xu Chuanqiang , Gao Lijia , Li Minghui , Liu Yiling , Qi Hongyan
Published: September 2022

Root-zone CO2 is essential for plant growth and metabolism. However, the partitioning and assimilation processes of CO2 absorbed by roots remain unclear in various parts of the oriental melon. We investigated the time at which root-zone CO2 enters the oriental melon root system, and its distribution in different parts of the plant, using 13C stable isotopic tracer experiments, as well as the effects of high root-zone CO2 on leaf carbon assimilation-related enzyme activities and gene expressions under 0.2%, 0.5% and 1% root-zone CO2 concentrations. The results showed that oriental melon roots could absorb CO2 and transport it quickly to the stems and leaves. The distribution of 13C in roots, stems and leaves increased with an increase in the labeled root-zone CO2 concentration, and the δ13C values in roots, stems and leaves increased initially, and then decreased with an increase in feeding time, reaching a peak at 24 h after 13C isotope labeling. The total accumulation of 13C in plants under the 0.5% and 1% 13CO2 concentrations was lower than that in the 0.2% 13CO2 treatment. However, the distributional proportion of 13C in leaves under 0.5% and 1% 13CO2 was significantly higher than that under the 0.2% CO2 concentration. Photosynthetic carbon assimilation-related enzyme activities and gene expressions in the leaves of oriental melon seedlings were inhibited after 9 days of high root-zone CO2 treatment. According to these results, oriental melon plants’ carbon distribution was affected by long-term high root-zone CO2, and reduced the carbon assimilation ability of the leaves. These findings provide a basis for the further quantification of the contribution of root-zone CO2 to plant communities in natural field conditions.
Impact of Freeze-Drying on Cord Blood (CB), Serum (S), and Platelet-Rich Plasma (CB-PRP) Preparations on Growth Factor Content and In Vitro Cell Wound Healing
Authors: Valente Sabrina , Ciavarella Carmen , Astolfi Gloria , Bergantin Elisa , Curti Nico , Buzzi Marina , Fontana Luigi , Versura Piera
Published: September 2022

Blood-based preparations are used in clinical practice for the treatment of several eye disorders. The aim of this study is to analyze the effect of freeze-drying blood-based preparations on the levels of growth factors and wound healing behaviors in an in vitro model. Platelet-rich plasma (PRP) and serum (S) preparations from the same Cord Blood (CB) sample, prepared in both fresh frozen (FF) and freeze-dried (FD) forms (and then reconstituted), were analyzed for EGF and BDNF content (ELISA Quantikine kit). The human MIO-M1 glial cell line (Moorfield/Institute of Ophthalmology, London, UK) was incubated with FF and FD products and evaluated for cell migration with scratch-induced wounding (IncuCyte S3 Essen BioScience), proliferation with cyclin A2 and D1 gene expression, and activation with vimentin and GFAP gene expression. The FF and FD forms showed similar concentrations of EGF and BDNF in both the S and PRP preparations. The wound healing assay showed no significant difference between the FF and FD forms for both S and PRP. Additionally, cell migration, proliferation, and activation did not appear to change in the FD forms compared to the FF ones. Our study showed that reconstituted FD products maintained the growth factor concentrations and biological properties of FF products and could be used as a functional treatment option.
An Axis between the Long Non-Coding RNA HOXA11-AS and NQOs Enhances Metastatic Ability in Oral Squamous Cell Carcinoma
Authors: Nakashima Chie , Fujiwara-Tani Rina , Mori Shiori , Kishi Shingo , Ohmori Hitoshi , Fujii Kiyomu , Mori Takuya , Miyagawa Yoshihiro , Yamamoto Kazuhiko , Kirita Tadaaki , Luo Yi , Kuniyasu Hiroki
Published: September 2022

Long non-coding RNAs (lncRNAs) play critical roles in human cancers. HOXA11 anti-sense RNA (HOXA11-AS) is an lncRNA belonging to the homeobox (HOX) gene cluster that promotes liver metastasis in human colon cancer. However, its role and mechanism of action in human oral squamous cell carcinoma (OSCC) are unclear. In this study, we investigated HOXA11-AS expression and function in human OSCC tissues and cell lines, as well as a mouse model of OSCC. Our analyses showed that HOXA11-AS expression in human OSCC cases correlates with lymph node metastasis, nicotinamide adenine dinucleotide (NAD)(P)H: quinone oxidoreductase 1 (NQO1) upregulation, and dihydronicotinamide riboside (NRH): quinone oxidoreductase 2 (NQO2) downregulation. Using the human OSCC cell lines HSC3 and HSC4, we demonstrate that HOXA11-AS promotes NQO1 expression by sponging microRNA-494. In contrast, HOXA11-AS recruits zeste homolog 2 (EZH2) to the NQO2 promoter to suppress its expression via the trimethylation of H3K27. The upregulation of NQO1 enzymatic activity by HOXA11-AS results in the consumption of flavin adenine dinucleotide (FAD), which reduces FAD-requiring glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity and suppresses glycolysis. However, our analyses show that lactic acid fermentation levels are preserved by glutaminolysis due to increased malic enzyme-1 expression, promoting enhanced proliferation, invasion, survival, and drug resistance. In contrast, suppression of NQO2 expression reduces the consumption of NRH via NQO2 enzymatic activity and increases NAD levels, which promotes enhanced stemness and metastatic potential. In mouse tumor models, knockdown of HOXA11-AS markedly suppressed tumor growth and lung metastasis. From these findings, targeting HOXA11-AS may strongly suppress high-grade OSCC by regulating both NQO1 and NQO2.
The Role of Vitamin D and Vitamin D Binding Protein in Chronic Liver Diseases
Authors: Pop Tudor Lucian , Sîrbe Claudia , Benţa Gabriel , Mititelu Alexandra , Grama Alina
Published: September 2022

Vitamin D (calciferol) is a fat-soluble vitamin that has a significant role in phospho-calcium metabolism, maintaining normal calcium levels and bone health development. The most important compounds of vitamin D are cholecalciferol (vitamin D3, or VD3) and ergocalciferol (vitamin D2, or VD2). Besides its major role in maintaining an adequate level of calcium and phosphate concentrations, vitamin D is involved in cell growth and differentiation and immune function. Recently, the association between vitamin D deficiency and the progression of fibrosis in chronic liver disease (CLD) was confirmed, given the hepatic activation process and high prevalence of vitamin D deficiency in these diseases. There are reports of vitamin D deficiency in CLD regardless of the etiology (chronic viral hepatitis, alcoholic cirrhosis, non-alcoholic fatty liver disease, primary biliary cirrhosis, or autoimmune hepatitis). Vitamin D binding protein (VDBP) is synthesized by the liver and has the role of binding and transporting vitamin D and its metabolites to the target organs. VDBP also plays an important role in inflammatory response secondary to tissue damage, being involved in the degradation of actin. As intense research during the last decades revealed the possible role of vitamin D in liver diseases, a deeper understanding of the vitamin D, vitamin D receptors (VDRs), and VDBP involvement in liver inflammation and fibrogenesis could represent the basis for the development of new strategies for diagnosis, prognosis, and treatment of liver diseases. This narrative review presents an overview of the evidence of the role of vitamin D and VDBP in CLD, both at the experimental and clinical levels.
Study on Enhancing the Corrosion Resistance and Photo-Thermal Antibacterial Properties of the Micro-Arc Oxidation Coating Fabricated on Medical Magnesium Alloy
Authors: Li Tianlu , Zhao Yun , Chen Minfang
Published: September 2022

Photo-thermal antibacterial properties have attracted much attention in the biomedical field because of their higher antibacterial efficiency. Through fabricating micro-arc oxidation coatings with different treating current densities set on a Mg-Zn-Ca alloy, the present study tried to systematically investigate and optimize the corrosion resistance and photo-thermal antibacterial properties of MAO coatings. The results indicated that different current densities had great influence on the corrosion resistance and photo-thermal property of the MAO coatings, and a current density at 30 A·dm−2 exhibited the best corrosion resistance, light absorption capacity at 808 nm, and photo-thermal capability, simultaneously with good antibacterial activity against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). This photo-thermal property of MAO coatings was probably related to the effect of current density on MgO content in the coating that could promote the separation of photo-generated electron carriers and hinder the recombination of photo-generated electron carriers and holes.
PSP-GNM: Predicting Protein Stability Changes upon Point Mutations with a Gaussian Network Model
Authors: Mishra Sambit Kumar
Published: September 2022

Understanding the effects of missense mutations on protein stability is a widely acknowledged significant biological problem. Genomic missense mutations may alter one or more amino acids, leading to increased or decreased stability of the encoded proteins. In this study, we describe a novel approach—Protein Stability Prediction with a Gaussian Network Model (PSP-GNM)—to measure the unfolding Gibbs free energy change (ΔΔG) and evaluate the effects of single amino acid substitutions on protein stability. Specifically, PSP-GNM employs a coarse-grained Gaussian Network Model (GNM) that has interactions between amino acids weighted by the Miyazawa–Jernigan statistical potential. We used PSP-GNM to simulate partial unfolding of the wildtype and mutant protein structures, and then used the difference in the energies and entropies of the unfolded wildtype and mutant proteins to calculate ΔΔG. The extent of the agreement between the ΔΔG calculated by PSP-GNM and the experimental ΔΔG was evaluated on three benchmark datasets: 350 forward mutations (S350 dataset), 669 forward and reverse mutations (S669 dataset) and 611 forward and reverse mutations (S611 dataset). We observed a Pearson correlation coefficient as high as 0.61, which is comparable to many of the existing state-of-the-art methods. The agreement with experimental ΔΔG further increased when we considered only those measurements made close to 25 °C and neutral pH, suggesting dependence on experimental conditions. We also assessed for the antisymmetry (ΔΔGreverse = −ΔΔGforward) between the forward and reverse mutations on the Ssym+ dataset, which has 352 forward and reverse mutations. While most available methods do not display significant antisymmetry, PSP-GNM demonstrated near-perfect antisymmetry, with a Pearson correlation of −0.97. PSP-GNM is written in Python and can be downloaded as a stand-alone code.
BAG3 Attenuates Ischemia-Induced Skeletal Muscle Necroptosis in Diabetic Experimental Peripheral Artery Disease
Authors: Mani Arul M. , Dhanabalan Karthik , Lamin Victor , Wong Thomas , Singh Madhu V. , Dokun Ayotunde O.
Published: September 2022

Peripheral artery disease (PAD) is characterized by impaired blood flow to the lower extremities, resulting in ischemic limb injuries. Individuals with diabetes and PAD typically have more severe ischemic limb injuries and limb amputations, but the mechanisms involved are poorly understood. Previously, we identified BAG3 as a gene within a mouse genetic locus termed limb salvage QTL1 on mouse chromosome 7 that determined the extent of limb necrosis following ischemic injury in C57Bl/6 mice. Whether BAG3 deficiency plays a role in the severe ischemic injury observed in diabetic PAD is not known. In vitro, we found simulated ischemia enhanced BAG3 expression in primary human skeletal muscle cells, whereas BAG3 knockdown increased necroptosis markers and decreased cell viability. In vivo, ischemic skeletal muscles from hind limbs of high-fat diet (HFD)-fed mice showed poor BAG3 expression compared to normal chow diet (NCD)-fed mice, and this was associated with increased limb amputations. BAG3 overexpression in ischemic skeletal muscles from hind limbs of HFD mice rescued limb amputation and improved autophagy, necroptosis, skeletal muscle function and regeneration. Therefore, BAG3 deficiency in ischemic skeletal muscles contributes to the severity of ischemic limb injury in diabetic PAD, likely through autophagy and necroptosis pathways.
Novel Dopamine Transporter Inhibitor, CE-123, Ameliorates Spatial Memory Deficits Induced by Maternal Separation in Adolescent Rats: Impact of Sex
Authors: Grochecki Pawel , Smaga Irena , Surowka Paulina , Marszalek-Grabska Marta , Kalaba Predrag , Dragacevic Vladimir , Kotlinska Patrycja , Filip Malgorzata , Lubec Gert , Kotlinska Jolanta H.
Published: September 2022

Maternal separation (MS) is a key contributor to neurodevelopmental disorders, including learning disabilities. To test the hypothesis that dopamine signaling is a major factor in this, an atypical new dopamine transporter (DAT) inhibitor, CE-123, was assessed for its potential to counteract the MS-induced spatial learning and memory deficit in male and female rats. Hence, neonatal rats (postnatal day (PND)1 to 21) were exposed to MS (180 min/day). Next, the acquisition of spatial learning and memory (Barnes maze task) and the expression of dopamine D1 receptor, dopamine transporter (DAT), and the neuronal GTPase, RIT2, which binds DAT in the vehicle-treated rats were evaluated in the prefrontal cortex and hippocampus in the adolescent animals. The results show that MS impairs the acquisition of spatial learning and memory in rats, with a more severe effect in females. Moreover, the MS induced upregulation of DAT and dopamine D1 receptors expression in the prefrontal cortex and hippocampus in adolescent rats. Regarding RIT2, the expression was decreased in the hippocampus for both the males and females, however, in the prefrontal cortex, reduction was found only in the females, suggesting that there are region-specific differences in DAT endocytic trafficking. CE-123 ameliorated the behavioral deficits associated with MS. Furthermore, it decreased the MS-induced upregulation of D1 receptor expression level in the hippocampus. These effects were more noted in females. Overall, CE-123, an atypical DAT inhibitor, is able to restore cognitive impairment and dopamine signaling in adolescent rats exposed to MS—with more evident effect in females than males.
Implementation of QbD Approach to the Development of Chromatographic Methods for the Determination of Complete Impurity Profile of Substance on the Preclinical and Clinical Step of Drug Discovery Studies
Authors: Gurba-Bryśkiewicz Lidia , Dawid Urszula , Smuga Damian A. , Maruszak Wioleta , Delis Monika , Szymczak Krzysztof , Stypik Bartosz , Moroz Aleksandra , Błocka Aleksandra , Mroczkiewicz Michał , Dubiel Krzysztof , Wieczorek Maciej
Published: September 2022

The purpose of this work was to demonstrate the use of the AQbD with the DOE approach to the methodical step-by-step development of a UHPLC method for the quantitative determination of the impurity profile of new CPL409116 substance (JAK/ROCK inhibitor) on the preclinical and clinical step of drug discovery studies. The critical method parameters (CMPs) have been tested extensively: the kind of stationary phase (8 different columns), pH of the aqueous mobile phase (2.6, 3.2, 4.0, 6.8), and start (20–25%) and stop (85–90%) percentage of organic mobile phase (ACN). The critical method attributes (CMAs) are the resolution between the peaks (≥2.0) and peak symmetry of analytes (≥0.8 and ≤1.8). In the screening step, the effects of different levels of CMPs on the CMAs were evaluated based on a full fractional design 22. The robustness tests were established from the knowledge space of the screening step and performed by application fractional factorial design 2(4−1). Method operable design region (MODR) was generated. The probability of meeting the specifications for the CMAs was calculated by Monte-Carlo simulations. In relation to literature such a complete AQbD approach including screening, optimization, and validation steps for the development of a new method for the quantitative determination of the full profile of nine impurities of an innovative pharmaceutical substance with the structure-based pre-development pointed out the novelty of our work. The final working conditions were as follows: column Zorbax Eclipse Plus C18, aqueous mobile phase 10 mM ± 1 mM aqueous solution of HCOOH, pH 2.6, 20% ± 1% of ACN at the start and 85% ± 1% of ACN at the end of the gradient, and column temperature 30 °C ± 2 °C. The method was validated in compliance with ICH guideline Q2(R1). The optimized method is specified, linear, precise, and robust. LOQ is on the reporting threshold level of 0.05% and LOD at 0.02% for all impurities.
Novel Anti-Neuroinflammatory Properties of a Thiosemicarbazone–Pyridylhydrazone Copper(II) Complex
Authors: Choo Xin Yi , McInnes Lachlan E. , Grubman Alexandra , Wasielewska Joanna M. , Belaya Irina , Burrows Emma , Quek Hazel , Martín Jorge Cañas , Loppi Sanna , Sorvari Annika , Rait Dzhessi , Powell Andrew , Duncan Clare , Liddell Jeffrey R. , Tanila Heikki , Polo Jose M. , Malm Tarja , Kanninen Katja M. , Donnelly Paul S. , White Anthony R.
Published: September 2022

Neuroinflammation has a major role in several brain disorders including Alzheimer’s disease (AD), yet at present there are no effective anti-neuroinflammatory therapeutics available. Copper(II) complexes of bis(thiosemicarbazones) (CuII(gtsm) and CuII(atsm)) have broad therapeutic actions in preclinical models of neurodegeneration, with CuII(atsm) demonstrating beneficial outcomes on neuroinflammatory markers in vitro and in vivo. These findings suggest that copper(II) complexes could be harnessed as a new approach to modulate immune function in neurodegenerative diseases. In this study, we examined the anti-neuroinflammatory action of several low-molecular-weight, charge-neutral and lipophilic copper(II) complexes. Our analysis revealed that one compound, a thiosemicarbazone–pyridylhydrazone copper(II) complex (CuL5), delivered copper into cells in vitro and increased the concentration of copper in the brain in vivo. In a primary murine microglia culture, CuL5 was shown to decrease secretion of pro-inflammatory cytokine macrophage chemoattractant protein 1 (MCP-1) and expression of tumor necrosis factor alpha (Tnf), increase expression of metallothionein (Mt1), and modulate expression of Alzheimer’s disease-associated risk genes, Trem2 and Cd33. CuL5 also improved the phagocytic function of microglia in vitro. In 5xFAD model AD mice, treatment with CuL5 led to an improved performance in a spatial working memory test, while, interestingly, increased accumulation of amyloid plaques in treated mice. These findings demonstrate that CuL5 can induce anti-neuroinflammatory effects in vitro and provide selective benefit in vivo. The outcomes provide further support for the development of copper-based compounds to modulate neuroinflammation in brain diseases.
The Perspective of Vitamin D on suPAR-Related AKI in COVID-19
Authors: Liao Tzu-Hsien , Wu Hsien-Chang , Liao Min-Tser , Hu Wan-Chung , Tsai Kuo-Wang , Lin Ching-Chieh , Lu Kuo-Cheng
Published: September 2022

The coronavirus disease 2019 (COVID-19) pandemic has claimed the lives of millions of people around the world. Severe vitamin D deficiency can increase the risk of death in people with COVID-19. There is growing evidence that acute kidney injury (AKI) is common in COVID-19 patients and is associated with poorer clinical outcomes. The kidney effects of SARS-CoV-2 are directly mediated by angiotensin 2-converting enzyme (ACE2) receptors. AKI is also caused by indirect causes such as the hypercoagulable state and microvascular thrombosis. The increased release of soluble urokinase-type plasminogen activator receptor (suPAR) from immature myeloid cells reduces plasminogen activation by the competitive inhibition of urokinase-type plasminogen activator, which results in low plasmin levels and a fibrinolytic state in COVID-19. Frequent hypercoagulability in critically ill patients with COVID-19 may exacerbate the severity of thrombosis. Versican expression in proximal tubular cells leads to the proliferation of interstitial fibroblasts through the C3a and suPAR pathways. Vitamin D attenuates the local expression of podocyte uPAR and decreases elevated circulating suPAR levels caused by systemic inflammation. This decrease preserves the function and structure of the glomerular barrier, thereby maintaining renal function. The attenuated hyperinflammatory state reduces complement activation, resulting in lower serum C3a levels. Vitamin D can also protect against COVID-19 by modulating innate and adaptive immunity, increasing ACE2 expression, and inhibiting the renin–angiotensin–aldosterone system. We hypothesized that by reducing suPAR levels, appropriate vitamin D supplementation could prevent the progression and reduce the severity of AKI in COVID-19 patients, although the data available require further elucidation.
Impact of Leptin on the Expression Profile of Macrophages during Mechanical Strain In Vitro
Authors: Paddenberg Eva , Osterloh Hannah , Jantsch Jonathan , Nogueira Andressa , Proff Peter , Kirschneck Christian , Schröder Agnes
Published: September 2022

Childhood obesity is a growing problem in industrial societies and associated with increased leptin levels in serum and salvia. Orthodontic treatment provokes pressure and tension zones within the periodontal ligament, where, in addition to fibroblasts, macrophages are exposed to these mechanical loadings. Given the increasing number of orthodontic patients with these conditions, insights into the effects of elevated leptin levels on the expression profile of macrophages during mechanical strain are of clinical interest. Therefore, the aim of this in vitro study was to assess the influence of leptin on the expression profile of macrophages during simulated orthodontic treatment. RAW264.7 macrophages were incubated with leptin and lipopolysaccharides (LPS) from Porphyromonas gingivalis (P. gingivalis) or with leptin and different types of mechanical strain (tensile, compressive strain). Expression of inflammatory mediators including tumor necrosis factor (TNF), Interleukin-1-B (IL1B), IL6, and prostaglandin endoperoxide synthase (PTGS2) was assessed by RT-qPCR, ELISAs, and immunoblot. Without additional mechanical loading, leptin increased Tnf, Il1b, Il6, and Ptgs2 mRNA in RAW264.7 macrophages by itself and after stimulation with LPS. However, in combination with tensile or compressive strain, leptin reduced the expression and secretion of these inflammatory factors. By itself and in combination with LPS from P. gingivalis, leptin has a pro-inflammatory effect. Both tensile and compressive strain lead to increased expression of inflammatory genes. In contrast to its effect under control conditions or after LPS treatment, leptin showed an anti-inflammatory phenotype after mechanical stress.
Interaction of miR-155 with Human Serum Albumin: An Atomic Force Spectroscopy, Fluorescence, FRET, and Computational Modelling Evidence
Authors: Botti Valentina , Cannistraro Salvatore , Bizzarri Anna Rita
Published: September 2022

This study investigated the interaction between Human Serum Albumin (HSA) and microRNA 155 (miR-155) through spectroscopic, nanoscopic and computational methods. Atomic force spectroscopy together with static and time-resolved fluorescence demonstrated the formation of an HSA/miR-155 complex characterized by a moderate affinity constant (KA in the order of 104 M−1). Förster Resonance Energy Transfer (FRET) experiments allowed us to measure a distance of (3.9 ± 0.2) nm between the lone HSA Trp214 and an acceptor dye bound to miR-155 within such a complex. This structural parameter, combined with computational docking and binding free energy calculations, led us to identify two possible models for the structure of the complex, both characterized by a topography in which miR-155 is located within two positively charged pockets of HSA. These results align with the interaction found for HSA and miR-4749, reinforcing the thesis that native HSA is a suitable miRNA carrier under physiological conditions for delivering to appropriate targets.
Gut Dysbiosis and Fecal Microbiota Transplantation in Autoimmune Diseases
Authors: Belvoncikova Paulina , Maronek Martin , Gardlik Roman
Published: September 2022

Gut microbiota dysbiosis has recently been reported in a number of clinical states, including neurological, psychiatric, cardiovascular, metabolic and autoimmune disorders. Yet, it is not completely understood how colonizing microorganisms are implicated in their pathophysiology and molecular pathways. There are a number of suggested mechanisms of how gut microbiota dysbiosis triggers or sustains extraintestinal diseases; however, none of these have been widely accepted as part of the disease pathogenesis. Recent studies have proposed that gut microbiota and its metabolites could play a pivotal role in the modulation of immune system responses and the development of autoimmunity in diseases such as rheumatoid arthritis, multiple sclerosis or type 1 diabetes. Fecal microbiota transplantation (FMT) is a valuable tool for uncovering the role of gut microbiota in the pathological processes. This review aims to summarize the current knowledge about gut microbiota dysbiosis and the potential of FMT in studying the pathogeneses and therapies of autoimmune diseases. Herein, we discuss the extraintestinal autoimmune pathologies with at least one published or ongoing FMT study in human or animal models.
BAT26 Only Microsatellite Instability with High Tumor Mutation Burden—A Rare Entity Associated with PTEN Protein Loss and High PD-L1 Expression
Authors: Kang So Young , Kim Deok Geun , Kim Kyoung-Mee
Published: September 2022

Detecting microsatellite instability (MSI) in advanced cancers is crucial for clinical decision-making, as it helps in identifying patients with differential treatment responses and prognoses. BAT26 is a highly sensitive MSI marker that defines the mismatch repair (MMR) status with high sensitivity and specificity. However, isolated BAT26-only instability is rare and has not been previously reported. Of the 6476 cases tested using pentaplex MSI polymerase chain reaction, we identified two BAT26-only instability cases (0.03%) in this study. The case #1 patient was diagnosed with endometrial adenocarcinoma without MMR germline mutations. The endometrial tumor showed BAT26-only instability, partial loss of MLH1/PMS2 protein expression, and a high programmed cell death ligand 1 (PD-L1) combined positive score (CPS = 8). The tumor exhibited a somatic phosphatase and tensin homolog (PTEN) R303P missense mutation and loss of the PTEN protein. On a comprehensive cancer panel sequencing with ≥500 genes, the tumor showed an MSI score of 11.38% and high tumor mutation burden (TMB) (19.5 mt/mb). The case #2 patient was diagnosed with colorectal carcinoma with proficient MMR and PTEN protein loss without PTEN alteration, as well as a high PD-L1 CPS (CPS = 10). A pathogenic KRAS A146T mutation was detected with an MSI score of 3.36% and high TMB (13 mt/mb). In conclusion, BAT26-only instability is very rare and associated with PTEN protein loss, high TMB, and a high PD-L1 score. Our results suggest that patients with BAT26-only instability may show good responses to immunotherapy.
New Insights into Ion Channels: Predicting hERG-Drug Interactions
Authors: Wempe Michael Fitzpatrick
Published: September 2022

Drug-induced long QT syndrome can be a very dangerous side effect of existing and developmental drugs. In this work, a model proposed two decades ago addressing the ion specificity of potassium channels is extended to the human ether-à-gogo gene (hERG). hERG encodes the protein that assembles into the potassium channel responsible for the delayed rectifier current in ventricular cardiac myocytes that is often targeted by drugs associated with QT prolongation. The predictive value of this model can guide a rational drug design decision early in the drug development process and enhance NCE (New Chemical Entity) retention. Small molecule drugs containing a nitrogen that can be protonated to afford a formal +1 charge can interact with hERG to prevent the repolarization of outward rectifier currents. Low-level ab initio calculations are employed to generate electronic features of the drug molecules that are known to interact with hERG. These calculations were employed to generate structure–activity relationships (SAR) that predict whether a small molecule drug containing a protonated nitrogen has the potential to interact with and inhibit the activity of the hERG potassium channels of the heart. The model of the mechanism underlying the ion specificity of potassium channels offers predictive value toward optimizing drug design and, therefore, minimizes the effort and expense invested in compounds with the potential for life-threatening inhibitory activity of the hERG potassium channel.
Hypochlorous Acid Chemistry in Mammalian Cells—Influence on Infection and Role in Various Pathologies
Authors: Andrés Celia María Curieses , Pérez de la Lastra José Manuel , Juan Celia Andrés , Plou Francisco J. , Pérez-Lebeña Eduardo
Published: September 2022

This review discusses the formation of hypochlorous acid HOCl and the role of reactive chlorinated species (RCS), which are catalysed by the enzyme myeloperoxidase MPO, mainly located in leukocytes and which in turn contribute to cellular oxidative stress. The reactions of RCS with various organic molecules such as amines, amino acids, proteins, lipids, carbohydrates, nucleic acids, and DNA are described, and an attempt is made to explain the chemical mechanisms of the formation of the various chlorinated derivatives and the data available so far on the effects of MPO, RCS and halogenative stress. Their presence in numerous pathologies such as atherosclerosis, arthritis, neurological and renal diseases, diabetes, and obesity is reviewed and were found to be a feature of debilitating diseases.
Human IRP1 Translocates to the Nucleus in a Cell-Specific and Iron-Dependent Manner
Authors: Gu Wen , Fillebeen Carine , Pantopoulos Kostas
Published: September 2022

Iron regulatory protein 1 (IRP1) is a bifunctional protein with mutually exclusive RNA-binding or enzymatic activities that depend on the presence of a 4Fe-4S cluster. While IRP1 is a well-established cytosolic protein, work in a Drosophila model suggested that it may also exhibit nuclear localization. Herein, we addressed whether mammalian IRP1 can likewise translocate to the nucleus. We utilized primary cells and tissues from wild type and Irp1−/− mice, as well as human cell lines and tissue biopsy sections. IRP1 subcellular localization was analyzed by Western blotting, immunofluorescence and immunohistochemistry. We did not detect presence of nuclear IRP1 in wild type mouse embryonic fibroblasts (MEFs), primary hepatocytes or whole mouse liver. However, we observed IRP1-positive nuclei in human liver but not ovary sections. Biochemical fractionation studies revealed presence of IRP1 in the nucleus of human Huh7 and HepG2 hepatoma cells, but not HeLa cervical cancer cells. Importantly, nuclear IRP1 was only evident in iron-replete cells and disappeared following pharmacological iron chelation. These data provide the first experimental evidence for nuclear IRP1 expression in mammals, which appears to be species- and cell-specific. Furthermore, they suggest that the nuclear translocation of IRP1 is mediated by an iron-dependent mechanism.
Screening of Induced Mutants Led to the Identification of Starch Biosynthetic Genes Associated with Improved Resistant Starch in Wheat
Authors: Irshad Ahsan , Guo Huijun , Ur Rehman Shoaib , Gu Jiayu , Wang Chaojie , Xiong Hongchun , Xie Yongdun , Zhao Shirong , Liu Luxiang
Published: September 2022

Several health benefits are obtained from resistant starch, also known as healthy starch. Enhancing resistant starch with genetic modification has huge commercial importance. The variation of resistant starch content is narrow in wheat, in relation to which limited improvement has been attained. Hence, there is a need to produce a wheat population that has a wide range of variations in resistant starch content. In the present study, stable mutants were screened that showed significant variation in the resistant starch content. A megazyme kit was used for measuring the resistant starch content, digestible starch, and total starch. The analysis of variance showed a significant difference in the mutant population for resistant starch. Furthermore, four diverse mutant lines for resistant starch content were used to study the quantitative expression patterns of 21 starch metabolic pathway genes; and to evaluate the candidate genes for resistant starch biosynthesis. The expression pattern of 21 starch metabolic pathway genes in two diverse mutant lines showed a higher expression of key genes regulating resistant starch biosynthesis (GBSSI and their isoforms) in the high resistant starch mutant lines, in comparison to the parent variety (J411). The expression of SBEs genes was higher in the low resistant starch mutants. The other three candidate genes showed overexpression (BMY, Pho1, Pho2) and four had reduced (SSIII, SBEI, SBEIII, ISA3) expression in high resistant starch mutants. The overexpression of AMY and ISA1 in the high resistant starch mutant line JE0146 may be due to missense mutations in these genes. Similarly, there was a stop_gained mutation for PHO2; it also showed overexpression. In addition, the gene expression analysis of 21 starch metabolizing genes in four different mutants (low and high resistant starch mutants) shows that in addition to the important genes, several other genes (phosphorylase, isoamylases) may be involved and contribute to the biosynthesis of resistant starch. There is a need to do further study about these new genes, which are responsible for the fluctuation of resistant starch in the mutants.
AtABCG14: A Long-Distance Root-to-Shoot Carrier of Cytokinin
Authors: Kaur Gurleen , Mishra Divya
Published: September 2022

Root-synthesized cytokinins regulate the growth, development, and stress responses of aboveground tissues and follow the transport route via xylem tissue. Arabidopsis ATP-binding cassette (ABC) transporter G family member 14 (AtABCG14) is involved in the xylem loading of root-synthesized cytokinins. However, the phloem unloading of root-synthesized cytokinin and shoot distribution have remained elusive. The recent study by Zhao et al., (2021) proved that the AtABCG14 protein mediates the phloem unloading of cytokinins through the apoplastic pathway indicating the AtABCG14 is a master regulator of shoot cytokinin distribution.
The Lyon Consensus Criteria for GERD Diagnosis in a Greek Population: The Clinical Impact and Changes in GERD Diagnosis in a Real-World, Retrospective Study
Authors: Voulgaris Theodoros , Lekakis Vasileios , Orfanidou Afroditi , Vlachogiannakos Jiannis , Kamberoglou Dimitrios , Papatheodoridis George , Karamanolis George
Published: September 2022

(1) Introduction/aim: Gastroesophageal reflux disease (GERD) affects 8–33% globally. The gold standard examination technique in diagnosing GERD is 24 h pHmetry ± impedance. Recently, new diagnostic criteria were introduced by the Lyon Consensus for GERD diagnosis. Our aim was to investigate the diagnostic yield of pHmetry + impedance using the Lyon Consensus criteria in a real-world study. (2) Patients and methods: Our study included 249 consecutive patients (M/F: 120/129, mean age 50 ± 15 years) who underwent 24 h pH+ impedance monitoring in our department, during a 5-year period. Epidemiological, endoscopic, clinical, and 24 h pH+ impedance data were retrospectively collected. (3) Results: Typical GERD symptoms were reported by 140/249 (56.2%) patients, whereas 99/249 (39.6%) patients reported various extraesophageal symptoms. Endoscopic findings supportive of GERD based on the Lyon Consensus were present in 42/185 (22.7%). An AET value of >6% was observed in 60/249 (24.1%). GERD diagnosis according to the Lyon Consensus criteria was set in 63/249 (25.3%) patients; a rate significantly lower than that observed by implementing the older criteria (32.1%), p < 0.001. In the multivariate analysis, the existence of endoscopic findings supportive of GERD diagnosis as defined by the Lyon Consensus (p = 0.036), a De Meester score of over 14.7, and the presence of typical GERD symptoms were correlated to GERD diagnosis (p < 0.001, respectively) using the criteria defined for pH–impedance monitoring. (4) Conclusions: Changes in the diagnostic criteria concerning the 24 h pH–impedance monitoring of GERD based on the Lyon Consensus led to a conclusive GERD diagnosis in approximately 25% of the patients. This rate of GERD diagnosis is reduced in comparison to the one confirmed with the use of previously established criteria.
Long-Term Outcomes of Same Patient Eyes Treated with Pars Plana Vitrectomy in One Eye and Conventional Treatment in the Other for Complications of Proliferative Diabetic Retinopathy
Authors: Berrocal Maria H. , Acaba-Berrocal Luis , Acaba Alexandra M.
Published: September 2022

The purpose of this study was to evaluate the long-term, real-world outcomes of pars plana vitrectomy (PPV) for complications of proliferative diabetic retinopathy. A retrospective review involving 64 patients with proliferative diabetic retinopathy that underwent PPV in their worse-seeing eye were followed for a minimum of 8 years. The fellow eye underwent conventional treatment. Patients were divided into two groups by age: patients younger than 50 years of age and patients older than 50. In the younger than 50 group, 89% of vitrectomized eyes had improved visual acuity (VA) while 3.6% had decreased VA. A total of 14% of vitrectomized eyes required additional laser and 11% required reoperations. In the conventional treatment eyes, 25% had improved VA while 68% had decreased VA (p < 0.05). A total of 72% required additional laser and 60% required PPV. In the older than 50 group, 86% of vitrectomized eyes had VA improvement and 3% had decreased VA. A total of 8% required laser and 8% required reoperations. In the conventional treatment eyes, 30% improved VA and 48% had decreased VA (p < 0.05). Additional procedures required included laser in 70% and PPV in 17%. In both age groups, eyes that underwent PPV had better final visual outcomes than eyes that received conventional treatment for PDR.
Biomarker-Guided Assessment of Acute Kidney Injury Phenotypes E among ST-Segment Elevation Myocardial Infarction Patients
Authors: Banai Ariel , Frydman Shir , Abu Katash Hytham , Stark Moshe , Goldiner Ilana , Banai Shmuel , Shacham Yacov
Published: September 2022

Recent practice guidelines recommended the use of new stress, functional, and damage biomarkers in clinical practice to prevent and manage acute kidney injury (AKI). Biomarkers are one of the tools used to define various AKI phenotypes and provide prognostic information regardless of an acute decline in renal function. We investigated the incidence and possible implications of AKI phenotypes among ST elevation myocardial infarction patient treated with primary coronary intervention. We included 281 patients with STEMI treated with PCI. Neutrophil gelatinase associated lipocalin (NGAL) was utilized to determine structural renal damage and functional AKI was determined using the KDIGO criteria. Patients were stratified into four AKI phenotypes: no AKI, subclinical AKI, hemodynamic AKI, and severe AKI. Patients were assessed for in-hospital adverse events (MACE). A total of 46 patients (44%) had subclinical AKI, 17 (16%) had hemodynamic AKI, and 42 (40%) had severe AKI. We observed a gradual and significant increase in the occurrence of MACE between the groups being highest among patients with severe AKI (10% vs. 19% vs. 29% vs. 43%; p < 0.001). In a multivariable regression model, any AKI phenotype was independently associated with MACE with an odds ratio of 4.15 (95% CI 2.1–8.3, p < 0.001,) for subclinical AKI, 4.51 (95% CI 1.61–12.69; p = 0.004) for hemodynamic AKI, and 12.9 (95% CI 5.59–30.1, p < 0.001) for severe AKI. In conclusion, among STEMI patients, AKI is a heterogeneous condition consisting of distinct phenotypes, addition of novel biomarkers may overcome the limitations of sCr-based AKI definitions to improve AKI phenotyping and direct potential therapies.
Patterns of Outpatient Phecodes Predating the Diagnosis of Systemic Lupus Erythematosus in Taiwanese Women
Authors: Lu Ming-Chi , Hsu Chia-Wen , Koo Malcolm
Published: September 2022

Shortening the time to diagnosis and initiating early treatment are imperative to improve outcomes in patients with systemic lupus erythematosus (SLE). The aim of this case-control study, based on the data from the Taiwan’s National Health Insurance Research Database (NHIRD), was to investigate the patterns of diagnoses of disease phenotypes in female patients with SLE up to eight years prior to its definitive diagnosis. The 547 cases were selected from the 2000–2012 NHIRD catastrophic illness datafile and frequency-matched with 2188 controls. The primary diagnosis based on the first ICD-9-CM code for each outpatient visit was converted to Phecodes. Separate regression models, based on least absolute shrinkage and selection operator (lasso) regularization, with seven different lag periods from 1–2 to 7–8 years, were conducted. Results showed that SLE was associated with 46 disease phenotypes in a lag period of 2–3 years, but fewer in other lag periods. A number of SLE-associated disease phenotypes, such as primary thrombocytopenia, thyroid diseases, Raynaud’s syndrome, renal disease, and several infectious diseases, occurred mainly in the first few years prior to SLE diagnosis. In conclusion, SLE should be suspected when the disease phenotypes identified in the present study occurred concomitantly.
Clinical Characteristics and Risk Factors of First-Ever Stroke in Young Adults: A Multicenter, Prospective Cohort Study
Authors: Jo Yea Jin , Kim Dae Hyun , Sohn Min Kyun , Lee Jongmin , Shin Yong-Il , Oh Gyung-Jae , Lee Yang-Soo , Joo Min Cheol , Lee So Young , Song Min-Keun , Han Junhee , Ahn Jeonghoon , Chang Won Hyuk , Kim Yun-Hee , Kim Deog Young
Published: September 2022

Stroke in young adults has catastrophic consequences and has increased in prevalence, contrary to the trends of most other diseases. This study aimed to determine the major characteristics and risk factors for stroke in younger adults compared with older adults. From the Korean Stroke Cohort for Functioning and Rehabilitation, 10,584 patients with first-ever stroke between August 2012 and March 2015 were enrolled retrospectively and divided into younger (age ≤ 45) and older groups (age > 45). The clinical characteristics and risk factors of stroke were compared between the younger and older groups. The younger group comprised 915 patients (8.6%). The proportion of hemorrhage strokes in the younger group (42.3%) was significantly higher than in the older group (20.0%) (p < 0.001). Obesity, current smoking, and heavy alcohol consumption were significantly more common risk factors in the younger group than in the older group for all stroke types, whereas hypertension, diabetes mellitus, hyperlipidemia, atrial fibrillation, and coronary heart disease were significantly more frequent in the older group (both p < 0.001). The major risk factors in the younger group may be lifestyle-related. Therefore, increasing awareness of lifestyle-related risk factors may be necessary to prevent stroke in young adults.
Factors Affecting Success and Survival of Small and Medium Enterprises in the Middle East
Authors: Ismail Albalushi Khalid , Naqshbandi M. Muzamil
Published: September 2022

SMEs are one of the leading solutions for reducing unemployment and poverty and boosting economic growth. Due to this, the determinants of the survival and success of SMEs have received increasing scrutiny in recent years. Empirical evidence has been uncovered in different countries. Enriching this growing body of evidence, this paper explores the internal and external factors that affect the survival and success of SMEs in the middle eastern country of Oman. We used a quantitative approach to collect the data by distributing a survey questionnaire among SME owners and prospective entrepreneurs. The survey was distributed in different industries throughout Oman. The findings, based on 344 responses, show that for SME survival and success, the education system needs intervention. Other areas of intervention include transforming Omani business culture, focusing on managerial skills, and improving the procedures required for establishing a business. These findings offer vital implications for Oman’s economy and for SME owners. The findings of this study can help policymakers make the appropriate interventions at various levels to enhance SME survival and success in Oman. The study also provides insights for existing and prospective entrepreneurs to bridge the skillset gaps to keep pace with ever-changing market demands.
Research on Manufacturers’ Referral Strategy Considering Store Brand Retailers and Traditional Retailers
Authors: Han Feiyan , Wang Herui , Lv Hongyu , Li Bo
Published: September 2022

It has become a common commercial phenomenon for retailers to establish their own brands. The manufacturer referral strategy is studied through a model which includes a manufacturer, a traditional retailer and a store brand retailer. We conduct research on the three cooperation methods of the manufacturer: “no information referral”, “exclusive referral” and “nonexclusive referral”. The equilibrium wholesale price, the manufacturer’s order quantity and the retailer’s own product output are studied by constructing game models, and the best referral cooperation choice between the manufacturer and the retailer is analysed according to their profit. The results show that the manufacturer’s referral level choice does not change the number of products, while the manufacturer’s market loss rate leads to a change in product order quantity among different choices. Under the combined effect of the market loss rate and the intensity of market competition, the store brand retailer will change the output decision of its own products. When the market loss rate meets a certain range, the manufacturer’s product sales can be maximized. For the manufacturer, any referral strategy is better than no referral strategy, and in most cases, the manufacturer prefers nonexclusive referrals. The traditional retailer is willing to accept the manufacturer’s referral cooperation, and the traditional retailer’s profit is better under the nonexclusive referrals; while most store brand retailers are willing to choose the nonexclusive referrals.
Membrane Separation Processes and Post-Combustion Carbon Capture: State of the Art and Prospects
Authors: Favre Eric
Published: September 2022

Membrane processes have been investigated for carbon capture for more than four decades. Important efforts have been more recently achieved for the development of advanced materials and, to a lesser extent, on process engineering studies. A state-of-the-art analysis is proposed with a critical comparison to gas absorption technology, which is still considered as the best available technology for this application. The possibilities offered by high-performance membrane materials (zeolites, Carbon Molecular Sieves, Metal Oxide Frameworks, graphenes, facilitated transport membranes, etc.) are discussed in combination to process strategies (multistage design, hybrid processes, energy integration). The future challenges and open questions of membranes for carbon capture are finally proposed.