bluuebunny/crossref_metadata_2025 huggingface dataset:
A Chloroplast-Localized Glucose-6-Phosphate Dehydrogenase Positively Regulates Stripe Rust Resistance in Wheat
Authors: Wei Xiaobo , Huang Xueling , Yang Weiling , Wang Xinran , Guan Tao , Kang Zhensheng , Liu Jie
Published: December 2022

Glucose-6-phosphate dehydrogenase (G6PDH), the rate-limiting enzyme of the pentose phosphate pathway (PPP), plays a pivotal role in plant stress responses. However, the function and mechanism of G6PDHs in crop plants challenged by fungal pathogens remain poorly understood. In this study, a wheat G6DPH gene responding to infection by Puccinia striiformis f. sp. tritici (Pst), designated TaG6PDH2, was cloned and functionally identified. TaG6PDH2 expression was significantly upregulated in wheat leaves inoculated with Pst or treated with abiotic stress factors. Heterologous mutant complementation and enzymatic properties indicate that TaG6PDH2 encodes a G6PDH protein. The transient expression of TaG6PDH2 in Nicotiana benthamiana leaves and wheat protoplasts revealed that TaG6PDH2 is a chloroplast-targeting protein. Silencing TaG6PDH2 via the barley stripe mosaic virus (BSMV)-induced gene silencing (VIGS) system led to compromised wheat resistance to the Pst avirulent pathotype CYR23, which is implicated in weakened H2O2 accumulation and cell death. In addition, TaG6PDH2 was confirmed to interact with the wheat glutaredoxin TaGrxS4. These results demonstrate that TaG6PDH2 endows wheat with increased resistance to stripe rust by regulating reactive oxygen species (ROS) production.
Comparative Analysis of Tumor-Associated microRNAs and Tetraspanines from Exosomes of Plasma and Ascitic Fluids of Ovarian Cancer Patients
Authors: Yunusova Natalia , Dzhugashvili Ekaterina , Yalovaya Alena , Kolomiets Larisa , Shefer Aleksei , Grigor’eva Alina , Tupikin Alexey , Kondakova Irina , Tamkovich Svetlana
Published: December 2022

Ovarian cancer (OC) is one of the most common and fatal types of gynecological cancer. In the early phase of OC detection, the current treatment and diagnostic methods are not efficient and sensitive enough. Therefore, it is crucial to explore the mechanisms of OC metastasis and discover valuable factors for early diagnosis of female cancers and novel therapeutic strategies for metastasis. Exosomes are known to be involved in the development, migration, and invasion of cancer cells, and their cargo could be useful for the non-invasive biopsy development. CD151- and Tspan8-positive exosomes are known to support the degradation of the extracellular matrix, and are involved in stroma remodeling, angiogenesis and cell motility, as well as the association of miR-24 and miR-101 with these processes. The objective of this study was to explore the relationship of these components of exosomal cargo, in patients with OC, to clarify the clinical significance of these markers in liquid biopsies. The levels of tetraspanins Tspan8+ and CD151+ exosomes were significantly higher in plasma exosomes of OC patients compared with healthy females (HFs). The relative levels of miR-24 and miR-101 in plasma exosomes of HFs were significantly higher than in plasma exosomes of OC patients, while the levels of these microRNAs in exosomes from plasma and ascites of ill females showed no difference. Our study revealed a strong direct correlation between the change in the ascites exosomes CD151+Tspan8+ subpopulation level and the expression levels of the ascites (R = 0.81, p < 0.05) and plasma exosomal miR-24 (R = 0.74, p < 0.05) in OC patients, which confirms the assumption that exosomal cargo act synergistically to increase cellular motility, affecting cellular processes and signaling. Bioinformatics analysis confirmed the involvement of CD151 and Tspan8 tetraspanins and genes controlled by miR-24-3p and miR-101 in signaling pathways, which are crucial for carcinogenesis, demonstrating that these tetraspanins and microRNAs are potential biomarkers for OC screening, and predictors of poor clinicopathological behavior in tumors.
Complementary Roles of Two DNA Protection Proteins from Deinococcus geothermalis
Authors: Bae Min K. , Shin Eunjung , Lee Sung-Jae
Published: December 2022

The roles of two interrelated DNA protection protein in starved cells (Dps)—putative Dps Dgeo_0257 and Dgeo_0281—as orthologous proteins to DrDps1 for DNA binding, protection, and metal ion sensing were characterised in a Deinococcus geothermalis strain. Dgeo_0257 exhibited high DNA-binding affinity and formed a multimeric structure but lacked the conserved amino acid sequence for ferroxidase activity. In contrast, the Dgeo_0281 (DgDps1) protein was abundant in the early exponential phase, had a lower DNA-binding activity than Dgeo_0257, and was mainly observed in its monomeric or dimeric forms. Electrophoretic mobility shift assays demonstrated that both purified proteins bound nonspecifically to DNA, and their binding ability was affected by certain metal ions. For example, in the presence of ferrous and ferric ions, neither Dgeo_0257 nor Dgeo_0281 could readily bind to DNA. In contrast, both proteins exhibited more stable DNA binding in the presence of zinc and manganese ions. Mutants in which the dps gene was disrupted exhibited higher sensitivity to oxidative stress than the wild-type strain. Furthermore, the expression levels of each gene showed an opposite correlation under H2O2 treatment conditions. Collectively, these findings indicate that the putative Dps Dgeo_0257 and DgDps1 from D. geothermalis are involved in DNA binding and protection in complementary interplay ways compared to known Dps.
TSLP as a Potential Therapy in the Treatment of CRLF2 B Cell Acute Lymphoblastic Leukemia
Authors: Alkashgari Hossam R. , Ruiz-Jimenez Caleb , Stoian Cornelia , Coats Jacqueline S. , Baez Ineavely , Chirshev Evgeny , Martinez Shannalee R. , Dovat Sinisa , Francis-Boyle Olivia L. , Casiano Carlos A. , Payne Kimberly J.
Published: December 2022

Cytokine receptor-like factor 2 B-cell acute lymphoblastic leukemia (CRLF2 B-ALL) is a high-risk subtype characterized by CRLF2 overexpression with poor survival rates in children and adults. CRLF2 and interleukin-7 receptor alpha (IL-7Rα) form a receptor for the cytokine thymic stromal lymphopoietin (TSLP), which induces JAK/STAT and PI3K/AKT/mTOR pathway signals. Previous studies from our group showed that low TSLP doses increased STAT5, AKT, and S6 phosphorylation and contributed to CRLF2 B-ALL cell survival. Here we investigated the role of TSLP in the survival and proliferation of CRLF2 B-ALL cells in vitro and in vivo. We hypothesized that high doses of TSLP increase CRLF2 signals and contribute to increased proliferation of CRLF2 B-ALL cells in vitro and in vivo. Interestingly, we observed the opposite effect. Specifically, high doses of TSLP induced apoptosis in human CRLF2 B-ALL cell lines in vitro, prevented engraftment of CRLF2 B-ALL cells, and prolonged the survival of +TSLP patient-derived-xenograft mice. Mechanistically, we showed that high doses of TSLP induced loss of its receptor and loss of CRLF2 signals in vitro. These results suggest that high doses of TSLP could be further investigated as a potential therapy for the treatment of CRLF2 B-ALL.
Half-Life Extension and Biodistribution Modulation of Biotherapeutics via Red Blood Cell Hitch-Hiking with Novel Anti-Band 3 Single-Domain Antibodies
Authors: Nguyen Toan D. , Bordeau Brandon M. , Zhang Yu , Mattle Anna G. , Balthasar Joseph P.
Published: December 2022

Small therapeutic proteins are receiving increased interest as therapeutic drugs; however, their clinical success has been limited due to their rapid elimination. Here, we report a half-life extension strategy via strategy via red blood cell red blood cell (RBC) hitch-hiking. This manuscript details the development and characterization of novel anti-RBC single-domain antibodies (sdAbs), their genetic fusion to therapeutic antibody fragments (TAF) as bispecific fusion constructs, and their influence on TAF pharmacokinetics and biodistribution. Several sdAbs specific to the band 3 antigen were generated via phage-display technology. Binding affinity to RBCs was assessed via flow cytometry. Affinity maturation via random mutagenesis was carried out to improve the binding affinity of the sdAbs. Bi-specific constructs were generated by fusing the anti-RBC sdAbs with anti-tissue necrosis factor alpha (TNF-α) TAF via the use of a glycine-serine flexible linker, and assessments for binding were performed via enzyme-linked immunosorbent assay and flow cytometry. Pharmacokinetics of anti-RBC sdAbs and fusion constructs were evaluated following intravenous bolus dosing in mice at a 1 mg/kg dose. Two RBC-binding sdAbs, RB12 and RE8, were developed. These two clones showed high binding affinity to human RBC with an estimated KD of 17.7 nM and 23.6 nM and low binding affinity to mouse RBC with an estimated KD of 335 nM and 528 nM for RB12 and RE8, respectively. Two derivative sdAbs, RMA1, and RMC1, with higher affinities against mouse RBC, were generated via affinity maturation (KD of 66.9 nM and 30.3 nM, respectively). Pharmacokinetic investigations in mice demonstrated prolonged circulation half-life of an anti-RBC-TNF-α bispecific construct (75 h) compared to a non-RBC binding control (1.3 h). In summary, the developed anti-RBC sdAbs and fusion constructs have demonstrated high affinity in vitro, and sufficient half-life extension in vivo.
A New Indicator to Differentiate Thyroid Follicular Inclusions in Cervical Lymph Nodes from Patients with Thyroid Cancer
Authors: Otsubo Chieko , Mussazhanova Zhanna , Kurohama Hirokazu , Shalgimbayeva Gulzira , Ueki Nozomi , Matsuoka Yuki , Madiyeva Madina , Sato Shinya , Yamashita Hiroyuki , Nakashima Masahiro
Published: December 2022

Nodal metastasis is crucial for determining the stage of well-differentiated thyroid cancer (WTC) in patients older than 55. Well-formed thyroid follicular inclusions (TFIs) are occasionally encountered in the cervical lymph nodes (LNs) of patients with papillary thyroid carcinoma (PTC), and it is difficult to determine whether they are true nodal metastases or ectopic thyroid tissues (ETT). This study aimed to elucidate the impact of the expression of the DNA damage response molecule TP53-binding protein 1 (53BP1) using immunofluorescence (IF) as a biomarker to differentiate TFIs in cervical LN by comparing the mutation analyses of primary thyroid cancers. The data demonstrated the necessity for the differential diagnosis of true metastases from ETT among TFIs in cervical LNs. PTC-like nuclear features using hematoxylin–eosin staining combined with immunohistochemistry for conventional biomarkers of PTC, including BRAFV600E protein, were most helpful in identifying metastatic follicular-patterned carcinomas. In conclusion, IF analysis of 53BP1 expression could be an excellent ancillary technique to distinguish metastatic carcinoma or ETT from TFIs in LNs, particularly in cases other than BRAFV600E-mutated PTC.
Salt-Induced Changes in Cytosolic pH and Photosynthesis in Tobacco and Potato Leaves
Authors: Pecherina Anna , Grinberg Marina , Ageyeva Maria , Zanegina Daria , Akinchits Elena , Brilkina Anna , Vodeneev Vladimir
Published: December 2022

Salinity is one of the most common factors limiting the productivity of crops. The damaging effect of salt stress on many vital plant processes is mediated, on the one hand, by the osmotic stress caused by large concentrations of Na+ and Cl− outside the root and, on the other hand, by the toxic effect of these ions loaded in the cell. In our work, the influence of salinity on the changes in photosynthesis, transpiration, water content and cytosolic pH in the leaves of two important crops of the Solanaceae family—tobacco and potato—was investigated. Salinity caused a decrease in photosynthesis activity, which manifested as a decrease in the quantum yield of photosystem II and an increase in non-photochemical quenching. Along with photosynthesis limitation, there was a slight reduction in the relative water content in the leaves and a decrease in transpiration, determined by the crop water stress index. Furthermore, a decrease in cytosolic pH was detected in tobacco and potato plants transformed by the gene of pH-sensitive protein Pt-GFP. The potential mechanisms of the salinity influence on the activity of photosynthesis were analyzed with the comparison of the parameters’ dynamics, as well as the salt content in the leaves.
Products of Bisphenol A Degradation Induce Cytotoxicity in Human Erythrocytes (In Vitro)
Authors: Makarova Katerina , Olchowik-Grabarek Ewa , Drabikowski Krzysztof , Kurkowiak Justyna , Zawada Katarzyna
Published: December 2022

The aim of this work has been to study the possible degradation path of BPA under the Fenton reaction, namely to determine the energetically favorable intermediate products and to compare the cytotoxicity of BPA and its intermediate products of degradation. The DFT calculations of the Gibbs free energy at M06-2X/6-311G(d,p) level of theory showed that the formation of hydroquinone was the most energetically favorable path in a water environment. To explore the cytotoxicity the erythrocytes were incubated with BPA and three intermediate products of its degradation, i.e., phenol, hydroquinone and 4-isopropylphenol, in the concentrations 5–200 μg/mL, for 1, 4 and 24 h. BPA induced the strongest hemolytic changes in erythrocytes, followed by hydroquinone, phenol and 4-isopropylphenol. In the presence of hydroquinone, the highest level of RONS was observed, whereas BPA had the weakest effect on RONS generation. In addition, hydroquinone decreased the level of GSH the most. Generally, our results suggest that a preferable BPA degradation path under a Fenton reaction should be controlled in order to avoid the formation of hydroquinone. This is applicable to the degradation of BPA during waste water treatment and during chemical degradation in sea water.
Ipconazole Disrupts Mitochondrial Homeostasis and Alters GABAergic Neuronal Development in Zebrafish
Authors: Lee Giyoung , Banik Amit , Eum Juneyong , Hwang Byung Joon , Kwon Seung-Hae , Kee Yun
Published: December 2022

Ipconazole, a demethylation inhibitor of fungal ergosterol biosynthesis, is widely used in modern agriculture for foliar and seed treatment, and is authorized for use in livestock feed. Waste from ipconazole treatment enters rivers and groundwater through disposal and rain, posing potential toxicity to humans and other organisms. Its metabolites remain stable under standard hydrolysis conditions; however, their neurodevelopmental toxicity is unknown. We investigated the potential neurodevelopmental toxicity of ipconazole pesticides in zebrafish (Danio rerio). Our behavioral monitoring demonstrated that the locomotive activity of ipconazole-exposed zebrafish larvae was reduced during early development, even when morphological abnormalities were undetected. Molecular profiling demonstrated that the mitochondrial-specific antioxidants, superoxide dismutases 1 and 2, and the genes essential for mitochondrial genome maintenance and functions were specifically reduced in ipconazole-treated (0.02 μg/mL) embryos, suggesting underlying ipconazole-driven oxidative stress. Consistently, ipconazole treatment substantially reduced hsp70 expression and increased ERK1/2 phosphorylation in a dose-dependent manner. Interrupted gad1b expression confirmed that GABAergic inhibitory neurons were dysregulated at 0.02 μg/mL ipconazole, whereas glutamatergic excitatory and dopaminergic neurons remained unaffected, resulting in an uncoordinated neural network. Additionally, ipconazole-treated (2 μg/mL) embryos exhibited caspase-independent cell death. This suggests that ipconazole has the potential to alter neurodevelopment by dysregulating mitochondrial homeostasis.
The MAOA rs979605 Genetic Polymorphism Is Differentially Associated with Clinical Improvement Following Antidepressant Treatment between Male and Female Depressed Patients
Authors: Chappell Kenneth , Colle Romain , Bouligand Jérôme , Trabado Séverine , Fève Bruno , Becquemont Laurent , Corruble Emmanuelle , Verstuyft Céline
Published: December 2022

Major depressive disorder (MDD) is the leading cause of disability worldwide. Treatment with antidepressant drugs (ATD), which target monoamine neurotransmitters including serotonin (5HT), are only modestly effective. Monoamine oxidase (MAO) metabolizes 5HT to 5-hydroxy indoleacetic acid (5HIAA). Genetic variants in the X-chromosome-linked MAO-encoding genes, MAOA and MAOB, have been associated with clinical improvement following ATD treatment in depressed patients. Our aim was to analyze the association of MAOA and MAOB genetic variants with (1) clinical improvement and (2) the plasma 5HIAA/5HT ratio in 6-month ATD-treated depressed individuals. Clinical (n = 378) and metabolite (n = 148) data were obtained at baseline and up to 6 months after beginning ATD treatment (M6) in patients of METADAP. Mixed-effects models were used to assess the association of variants with the Hamilton Depression Rating Scale (HDRS) score, response and remission rates, and the plasma 5HIAA/5HT ratio. Variant × sex interactions and dominance terms were included to control for X-chromosome-linked factors. The MAOA rs979605 and MAOB rs1799836 polymorphisms were analyzed. The sex × rs979605 interaction was significantly associated with the HDRS score (p = 0.012). At M6, A allele-carrying males had a lower HDRS score (n = 24, 10.9 ± 1.61) compared to AA homozygous females (n = 14, 18.1 ± 1.87; p = 0.0067). The rs1799836 polymorphism was significantly associated with the plasma 5HIAA/5HT ratio (p = 0.018). Overall, CC/C females/males had a lower ratio (n = 44, 2.18 ± 0.28) compared to TT/T females/males (n = 60, 2.79 ± 0.27; p = 0.047). The MAOA rs979605 polymorphism, associated with the HDRS score in a sex-dependent manner, could be a useful biomarker for the response to ATD treatment.
Genome Editing Using Cas9 Ribonucleoprotein Is Effective for Introducing PDGFRA Variant in Cultured Human Glioblastoma Cell Lines
Authors: Hamada Taiji , Yokoyama Seiya , Akahane Toshiaki , Matsuo Kei , Tanimoto Akihide
Published: December 2022

Many variants of uncertain significance (VUS) have been detected in clinical cancer cases using next-generation sequencing-based cancer gene panel analysis. One strategy for the elucidation of VUS is the functional analysis of cultured cancer cell lines that harbor targeted gene variants using genome editing. Genome editing is a powerful tool for creating desired gene alterations in cultured cancer cell lines. However, the efficiency of genome editing varies substantially among cell lines of interest. We performed comparative studies to determine the optimal editing conditions for the introduction of platelet-derived growth factor receptor alpha (PDGFRA) variants in human glioblastoma multiforme (GBM) cell lines. After monitoring the copy numbers of PDGFRA and the expression level of the PDGFRα protein, four GBM cell lines (U-251 MG, KNS-42, SF126, and YKG-1 cells) were selected for the study. To compare the editing efficiency in these GBM cell lines, the modes of clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein 9 (Cas9) delivery (plasmid vs. ribonucleoprotein (RNP)), methods of transfection (lipofection vs. electroporation), and usefulness of cell sorting were then evaluated. Herein, we demonstrated that electroporation-mediated transfer of Cas9 with single-guide RNA (Cas9 RNP complex) could sufficiently edit a target nucleotide substitution, irrespective of cell sorting. As the Cas9 RNP complex method showed a higher editing efficiency than the Cas9 plasmid lipofection method, it was the optimal method for single-nucleotide editing in human GBM cell lines under our experimental conditions.
Influence of Two Widely Used Solvents, Ethanol and Dimethyl Sulfoxide, on Human Sperm Parameters
Authors: Bisconti Marie , Grosjean Philippe , Arcolia Vanessa , Simon Jean-François , Hennebert Elise
Published: December 2022

To study mechanisms involved in fertility, many experimental assays are conducted by incubating spermatozoa in the presence of molecules dissolved in solvents such as ethanol (EtOH) or dimethyl sulfoxide (DMSO). Although a vehicle control group is usually included in such studies, it does not allow to evaluate the intrinsic effect of the solvent on sperm parameters and its potential influence on the outcome of the experiment. In the present study, we incubated human spermatozoa for 4 h in a capacitation medium in the absence or the presence of different concentrations of EtOH and DMSO (0.1, 0.5, 1.0, and 2.0%) to assess the impact of these solvents on sperm motility, vitality, capacitation, and acrosome integrity. The presence of statistically significant relationships between increasing solvent concentrations and the investigated parameters was assessed using linear mixed models. A significant effect was observed with both solvents for total and progressive sperm motilities. We also evaluated the effect of time for these parameters and showed that the influence of the solvents was stable between 0 and 4 h, indicating an almost direct impact of the solvents. While EtOH did not influence sperm vitality and acrosome integrity, a significant effect of increasing DMSO concentrations was observed for these parameters. Finally, regarding capacitation, measured via phosphotyrosine content, although a dose-dependent effect was observed with both solvents, the statistical analysis did not allow to precisely evaluate the intensity of the effect. Based on the results obtained in the present study, and the corresponding linear mixed models, we calculated the concentration of both solvents which would result in a 5% decline in sperm parameters. For EtOH, these concentrations are 0.9, 0.7, and 0.3% for total motility, progressive motility, and capacitation, respectively, while for DMSO they are 1.5, 1.1, >2, 0.3 and >2% for total motility, progressive motility, vitality, capacitation, and acrosome integrity, respectively. We recommend using solvent concentrations below these values to dissolve molecules used to study sperm function in vitro, to limit side effects.
Correction: Chen et al. The Chemerin/CMKLR1 Axis Is Involved in the Recruitment of Microglia to Aβ Deposition through p38 MAPK Pathway. Int. J. Mol. Sci. 2022, 23, 9041
Authors: Chen Yanqing , Liu Zhen , Gong Ping , Zhang Haibo , Chen Yijun , Yao Songquan , Li Wei , Zhang Yan , Yu Yang
Published: December 2022

In the original publication [...]
The Role of Nucleases Cleaving TLR3, TLR7/8 and TLR9 Ligands, Dicer RNase and miRNA/piRNA Proteins in Functional Adaptation to the Immune Escape and Xenophagy of Prostate Cancer Tissue
Authors: Kocic Gordana , Hadzi-Djokic Jovan , Colic Miodrag , Veljkovic Andrej , Tomovic Katarina , Roumeliotis Stefanos , Smelcerovic Andrija , Liakopoulos Vassilios
Published: December 2022

The prototypic sensors for the induction of innate and adaptive immune responses are the Toll-like receptors (TLRs). Unusually high expression of TLRs in prostate carcinoma (PC), associated with less differentiated, more aggressive and more propagating forms of PC, changed the previous paradigm about the role of TLRs strictly in immune defense system. Our data reveal an entirely novel role of nucleic acids-sensing Toll-like receptors (NA-TLRs) in functional adaptation of malignant cells for supply and digestion of surrounding metabolic substrates from dead cells as specific mechanism of cancer cells survival, by corresponding ligands accelerated degradation and purine/pyrimidine salvage pathway. The spectrophotometric measurement protocols used for the determination of the activity of RNases and DNase II have been optimized in our laboratory as well as the enzyme-linked immunosorbent method for the determination of NF-κB p65 in prostate tissue samples. The protocols used to determine Dicer RNase, AGO2, TARBP2 and PIWIL4 were based on enzyme-linked immunosorbent assay. The amount of pre-existing acid-soluble oligonucleotides was measured and expressed as coefficient of absorbance. The activities of acid DNase II and RNase T2, and the activities of nucleases cleaving TLR3, TLR7/8 and TLR9 ligands (Poly I:C, poly U and unmethylated CpG), increased several times in PC, compared to the corresponding tumor adjacent and control tissue, exerting very high sensitivity and specificity of above 90%. Consequently higher levels of hypoxanthine and NF-κB p65 were reported in PC, whereas the opposite results were observed for miRNA biogenesis enzyme (Dicer RNase), miRNA processing protein (TARB2), miRNA-induced silencing complex protein (Argonaute-AGO) and PIWI-interacting RNAs silence transposon. Considering the crucial role of purine and pyrimidine nucleotides as energy carriers, subunits of nucleic acids and nucleotide cofactors, future explorations will be aimed to design novel anti-cancer immune strategies based on a specific acid endolysosomal nuclease inhibition.
Influence of the Peripheral Nervous System on Murine Osteoporotic Fracture Healing and Fracture-Induced Hyperalgesia
Authors: Wank Isabel , Niedermair Tanja , Kronenberg Daniel , Stange Richard , Brochhausen Christoph , Hess Andreas , Grässel Susanne
Published: December 2022

Osteoporotic fractures are often linked to persisting chronic pain and poor healing outcomes. Substance P (SP), α-calcitonin gene-related peptide (α-CGRP) and sympathetic neurotransmitters are involved in bone remodeling after trauma and nociceptive processes, e.g., fracture-induced hyperalgesia. We aimed to link sensory and sympathetic signaling to fracture healing and fracture-induced hyperalgesia under osteoporotic conditions. Externally stabilized femoral fractures were set 28 days after OVX in wild type (WT), α-CGRP- deficient (α-CGRP −/−), SP-deficient (Tac1−/−) and sympathectomized (SYX) mice. Functional MRI (fMRI) was performed two days before and five and 21 days post fracture, followed by µCT and biomechanical tests. Sympathectomy affected structural bone properties in the fracture callus whereas loss of sensory neurotransmitters affected trabecular structures in contralateral, non-fractured bones. Biomechanical properties were mostly similar in all groups. Both nociceptive and resting-state (RS) fMRI revealed significant baseline differences in functional connectivity (FC) between WT and neurotransmitter-deficient mice. The fracture-induced hyperalgesia modulated central nociception and had robust impact on RS FC in all groups. The changes demonstrated in RS FC in fMRI might potentially be used as a bone traumata-induced biomarker regarding fracture healing under pathophysiological musculoskeletal conditions. The findings are of clinical importance and relevance as they advance our understanding of pain during osteoporotic fracture healing and provide a potential imaging biomarker for fracture-related hyperalgesia and its temporal development. Overall, this may help to reduce the development of chronic pain after fracture thereby improving the treatment of osteoporotic fractures.
Functional Characterization of the Cystine-Rich-Receptor-like Kinases (CRKs) and Their Expression Response to Sclerotinia sclerotiorum and Abiotic Stresses in Brassica napus
Authors: Sarwar Rehman , Li Lei , Yu Jiang , Zhang Yijie , Geng Rui , Meng Qingfeng , Zhu Keming , Tan Xiao-Li
Published: December 2022

Cysteine-rich receptor-like kinases (CRKs) are transmembrane proteins that bind to the calcium ion to regulate stress-signaling and plant development-related pathways, as indicated by several pieces of evidence. However, the CRK gene family hasn’t been inadequately examined in Brassica napus. In our study, 27 members of the CRK gene family were identified in Brassica napus, which are categorized into three phylogenetic groups and display synteny relationship to the Arabidopsis thaliana orthologs. All the CRK genes contain highly conserved N-terminal PKINASE domain; however, the distribution of motifs and gene structure were variable conserved. The functional divergence analysis between BnaCRK groups indicates a shift in evolutionary rate after duplication events, demonstrating that BnaCRKs might direct a specific function. RNA-Seq datasets and quantitative real-time PCR (qRT-PCR) exhibit the complex expression profile of the BnaCRKs in plant tissues under multiple stresses. Nevertheless, BnaA06CRK6-1 and BnaA08CRK8 from group B were perceived to play a predominant role in the Brassica napus stress signaling pathway in response to drought, salinity, and Sclerotinia sclerotiorum infection. Insights gained from this study improve our knowledge about the Brassica napus CRK gene family and provide a basis for enhancing the quality of rapeseed.
Dosimetry Effects Due to the Presence of Fe Nanoparticles for Potential Combination of Hyperthermic Cancer Treatment with MRI-Based Image-Guided Radiotherapy
Authors: Gayol Amiel , Malano Francisco , Ribo Montenovo Clara , Pérez Pedro , Valente Mauro
Published: December 2022

Nanoparticles have proven to be biocompatible and suitable for many biomedical applications. Currently, hyperthermia cancer treatments based on Fe nanoparticle infusion excited by alternating magnetic fields are commonly used. In addition to this, MRI-based image-guided radiotherapy represents, nowadays, one of the most promising accurate radiotherapy modalities. Hence, assessing the feasibility of combining both techniques requires preliminary characterization of the corresponding dosimetry effects. The present work reports on a theoretical and numerical simulation feasibility study aimed at pointing out preliminary dosimetry issues. Spatial dose distributions incorporating magnetic nanoparticles in MRI-based image-guided radiotherapy have been obtained by Monte Carlo simulation approaches accounting for all relevant radiation interaction properties as well as charged particles coupling with strong external magnetic fields, which are representative of typical MRI-LINAC devices. Two main effects have been evidenced: local dose enhancement (up to 60% at local level) within the infused volume, and non-negligible changes in the dose distribution at the interfaces between different tissues, developing to over 70% for low-density anatomical cavities. Moreover, cellular uptakes up to 10% have been modeled by means of considering different Fe nanoparticle concentrations. A theoretical temperature-dependent model for the thermal enhancement ratio (TER) has been used to account for radiosensitization due to hyperthermia. The outcomes demonstrated the reliability of the Monte Carlo approach in accounting for strong magnetic fields and mass distributions from patient-specific anatomy CT scans to assess dose distributions in MRI-based image-guided radiotherapy combined with magnetic nanoparticles, while the hyperthermic radiosensitization provides further and synergic contributions.
Toxicity of Different Types of Surfactants via Cellular and Enzymatic Assay Systems
Authors: Sutormin Oleg S. , Kolosova Elizaveta M. , Torgashina Irina G. , Kratasyuk Valentina A. , Kudryasheva Nadezhda S. , Kinstler Julia S. , Stom Devard I.
Published: December 2022

Surfactants have a widespread occurrence, not only as household detergents, but also in their application in industry and medicine. There are numerous bioassays for assessing surfactant toxicity, but investigations of their impact on biological systems at the molecular level are still needed. In this paper, luminous marine bacteria and their coupled NAD(P)H:FMN-oxidoreductase + luciferase (Red + Luc) enzyme system was applied to examine the effects of different types of surfactants, including cationic cetyltrimethylammonium bromide (CTAB), non-ionic polyoxyethylene 20 sorbitan monooleate (Tween 80) and anionic sodium lauryl sulfate (SLS), and to assess whether the Red + Luc enzyme system can be used as a more sensitive indicator of toxicity. It was shown that the greatest inhibitory effect of the surfactants on the activity of luminous bacteria and the Red + Luc enzyme system was in the presence of SLS samples. The calculated IC50 and EC50 values of SLS were 10−5 M and 10−2 M for the enzymatic and cellular assay systems, respectively. The results highlight the benefits of using the enzymatic assay system in ecotoxicology as a tool for revealing surfactant effects on intracellular proteins if the cellular membrane is damaged under a long-term exposure period in the presence of the surfactants. For this purpose, the bioluminescent enzyme-inhibition-based assay could be used as an advanced research tool for the evaluation of surfactant toxicity at the molecular level of living organisms due to its technical simplicity and rapid response time.
Screening and Druggability Analysis of Marine Active Metabolites against SARS-CoV-2: An Integrative Computational Approach
Authors: Murugesan Selvakumar , Ragavendran Chinnasamy , Ali Amir , Arumugam Velusamy , Lakshmanan Dinesh Kumar , Palanichamy Palanikumar , Venkatesan Manigandan , Kamaraj Chinnaperumal , Luna-Arias Juan Pedro , Fabián Fernández-Luqueño , Khan Safir Ullah , Mashwani Zia ur-Rehman , Younas Muhammad
Published: December 2022

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections have triggered a recent pandemic of respiratory disease and affected almost every country all over the world. A large amount of natural bioactive compounds are under clinical investigation for various diseases. In particular, marine natural compounds are gaining more attention in the new drug development process. The present study aimed to identify potential marine-derived inhibitors against the target proteins of COVID-19 using a computational approach. Currently, 16 marine clinical-level compounds were selected for computational screening against the 4 SARS-CoV-2 main proteases. Computational screening resulted from the best drug candidates for each target based on the binding affinity scores and amino acid interactions. Among these, five marine-derived compounds, namely, chrysophaentin A (−6.6 kcal/mol), geodisterol sulfates (−6.6 kcal/mol), hymenidin (−6.4 kcal/mol), plinabulin (−6.4 kcal/mol), and tetrodotoxin (−6.3 kcal/mol) expressed minimized binding energy and molecular interactions, such as covalent and hydrophobic interactions, with the SARS CoV-2 main protease. Using molecular dynamic studies, the root-mean-square deviation (RMSD), root-mean-square fluctuation (RMSF), radius of gyration (ROG), and hydrogen bond (H-Bond) values were calculated for the SARS-CoV-2 main protease with a hymenidin docked complex. Additionally, in silico drug-likeness and pharmacokinetic property assessments of the compounds demonstrated favorable druggability. These results suggest that marine natural compounds are capable of fighting SARS-CoV-2. Further in vitro and in vivo studies need to be carried out to confirm their inhibitory potential.
Basketball Action Recognition Method of Deep Neural Network Based on Dynamic Residual Attention Mechanism
Authors: Xiao Jiongen , Tian Wenchun , Ding Liping
Published: December 2022

Aiming at the problem that the features extracted from the original C3D (Convolutional 3D) convolutional neural network(C3D) were insufficient, and it was difficult to focus on keyframes, which led to the low accuracy of basketball players’ action recognition; hence, a basketball action recognition method of deep neural network based on dynamic residual attention mechanism was proposed. Firstly, the traditional C3D is improved to a dynamic residual convolution network to extract sufficient feature information. Secondly, the extracted feature information is selected by the improved attention mechanism to obtain the key video frames. Finally, the algorithm is compared with the traditional C3D in order to demonstrate the advance and applicability of the algorithm. Experimental results show that this method can effectively recognize basketball posture, and the average accuracy of posture recognition is more than 97%.
An Efficient and Simple Method for Collecting Haemolymph of Cerambycidae (Insecta: Coleoptera) Adults
Authors: Niu Yiming , Zhao Yuxuan , Shi Fengming , Li Meng , Zhang Sainan , Yang Jinglin , Zong Shixiang , Tao Jing
Published: December 2022

Cerambycid beetles (Cerambycidae) are major forest pests, posing a serious threat to the security of forest resources worldwide. Extensive research has focused on the control of cerambycid beetles from physiological and biochemical perspectives. Despite the important roles of insect haemolymph in physiological processes, efficient collection methods for Cerambycidae are lacking. For the efficient and easy collection of large amounts of pure haemolymph from adult cerambycid beetles, a new method, named net centrifugation, was developed. Three species of cerambycid beetles with large differences in size, Anoplophora chinensis, Monochamus saltuarius and Saperda populnea, were selected for the study. Haemolymph was collected by the newly developed net centrifugation method—in which an inner nylon net is used during centrifugation under optimised conditions, and a relatively small wound is generated on the insect—as well as the traditional tearing method and double centrifugation method. Among the three methods evaluated, the net centrifugation method caused the least damage to cerambycid beetles during the whole operation. This method resulted in the most haemolymph from a single beetle, with the lowest turbidity, mostly pure haemocytes in the precipitate, the clearest haemolymph smears by microscopy and the highest quality of RNA extracted from haemocytes. The net centrifugation method has a high collection efficiency, providing important technical support for haemolymph extraction and entomological research.
Labial Sensory Organs of Two Leptoglossus Species (Hemiptera: Coreidae): Their Morphology and Supposed Function
Authors: Taszakowski Artur , Masłowski Adrian , Brożek Jolanta
Published: December 2022

A detailed description of the labial sensory organs of Leptoglossus occidentalis Heidemann, 1910 and L. zonatus (Dallas, 1852) (Hemiptera: Heteroptera: Coreidae) is presented. The detailed morphology, location, and distribution of different sensilla types on mouthparts were investigated and shown in micrographs taken with a scanning electron microscope. Nine types of aporous sensilla, and three uniporous sensilla were found. The possible functions of these sensilla as well as similarities and differences between the mouthparts of Leptoglossus and those of other terrestrial Heteroptera are discussed. The tip of the labium constitutes a functional “touch and taste area”, combining the chemosensitivity of uniporous sensilla P1–P3 and the mechanoreceptivity of A8 and A9 hair-like sensilla. A set of two cone-like chemosensilla types (9 + 2) was found on each lateral lobe of the labial tip. Literature analysis showed that such a set of cone-like sensory organs on the labial tip may be common in terrestrial Heteroptera. This observation confirms that the number and arrangement of sensilla is conservative and can be important in diagnosing taxa at various levels and in phylogenetic studies based on morphology.
Does Frenotomy Modify Upper Airway Collapse in OSA Adult Patients? Case Report and Systematic Review
Authors: Correa Eduardo J. , O’Connor-Reina Carlos , Rodríguez-Alcalá Laura , Benjumea Felipe , Casado-Morente Juan Carlos , Baptista Peter M. , Casale Manuele , Moffa Antonio , Plaza Guillermo
Published: December 2022

Ankyloglossia (tongue-tie) is a condition of the oral cavity in which an abnormally short lingual frenulum affects the tongue’s mobility. Literature on the correlation between ankyloglossia and obstructive sleep apnea (OSA) is scarce. The main objective of this study was to report our preliminary experience in adult OSA patients before and after ankyloglossia treatment, using drug-induced sleep endoscopy (DISE) to evaluate the upper airway modifications resulting after treatment, and to present a systematic review of the impact of ankyloglossia and its treatment on OSA adults. We found that, after frenotomy, regarding the DISE findings, and according to the VOTE classification, two of the three patients showed an improvement in tongue level, from 2A-P (complete anteroposterior collapse) to 1ap (partial anteroposterior collapse). The third patient showed no changes in his UA after frenotomy, neither worsening nor showing improvement. Thus, the results of this study suggest that frenotomy in OSA patients with ankyloglossia could reduce tongue collapse, probably by allowing the tongue to take into the physiological position in the oral cavity. These patients should undergo speech therapy and oropharyngeal exercises prior to any surgical procedure, in order to avoid glossoptosis and to improve the quality of life and sleep apnea results.
New Challenges in Sexuality and Sexual Dysfunction
Authors: Montejo Angel L.
Published: December 2022

Human sexuality constitutes not only a basic need but also a right that significantly enriches interpersonal relationships, providing mutual satisfaction and pleasure [...]
The Association of Placental Abruption and Pediatric Neurological Outcome: A Systematic Review and Meta-Analysis
Authors: Oltean Irina , Rajaram Ajay , Tang Ken , MacPherson James , Hondonga Tadiwanashe , Rishi Aanchal , Toltesi Regan , Gowans Rachel , Jahangirnia Ashkan , Nasr Youssef , Lawrence Sarah , El Demellawy Dina
Published: December 2022

Placental histopathology provides insights, or “snapshots”, into relevant antenatal factors that could elevate the risk of perinatal brain injury. We present a systematic review and meta-analysis comparing frequencies of adverse neurological outcomes in infants born to women with placental abruption versus without abruption. Records were sourced from MEDLINE, Embase, and the CENTRAL Trials Registry from 1946 to December 2019. Studies followed the PRISMA guidelines and compared frequencies of neurodevelopmental morbidities in infants born to pregnant women with placental abruption (exposure) versus women without placental abruption (comparator). The primary endpoint was cerebral palsy. Periventricular and intraventricular (both severe and any grades of IVH) and any histopathological neuronal damage were the secondary endpoints. Study methodologic quality was assessed by the Ottawa–Newcastle scale. Estimated odds ratios (OR) and hazards ratio (HR) were derived according to study design. Data were meta-analyzed using a random effects model expressed as pooled effect sizes and 95% confidence intervals. We included eight observational studies in the review, including 1245 infants born to women with placental abruption. Results of the random effects meta-analysis show that the odds of infants born to pregnant women with placental abruption who experience cerebral palsy is higher than in infants born to pregnant women without placental abruption (OR 5.71 95% CI (1.17, 27.91); I2 = 84.0%). There is no statistical difference in the odds of infants born to pregnant women with placental abruption who experience severe IVH (grade 3+) (OR 1.20 95% CI (0.46, 3.11); I2 = 35.8%) and any grade of IVH (OR 1.20 95% CI (0.62, 2.32); I2 = 32.3%) vs. women without placental abruption. There is no statistically significant difference in the odds of infants born to pregnant women with placental abruption who experience PVL vs. pregnant women without placental abruption (OR 6.51 95% CI (0.94, 45.16); I2 = 0.0%). Despite our meta-analysis suggesting increased odds of cerebral palsy in infants born to pregnant women with placental abruption versus without abruption, this finding should be interpreted cautiously, given high heterogeneity and overall poor quality of the included studies.
Cross-Sectional Study on Lateral Skull Radiographs to Design a New Nasopharyngeal Swab for Simplified COVID-19 and Respiratory Infections Diagnostic Testing in Children
Authors: Goracci Cecilia , Volpe Alessandra , Salerni Lorenzo , Paolini Elisabetta , Vichi Alessandro , Franchi Lorenzo
Published: December 2022

Nasopharyngeal swab sample collection is the first-line testing method for diagnosing COVID-19 infection and other respiratory infections. Current information on how to properly perform nasopharyngeal swabbing in children is largely defective. This study aimed at collecting nostril to nasopharynx distance measurements on lateral skull radiographs of children and adolescents to design a nasopharyngeal swab meant to standardize and facilitate the sample collection procedure. A total of 323 cephalograms of 152 male and 171 female children aged 4–14 years taken for orthodontic reasons were selected. On each cephalogram, the shortest distance between the most anterosuperior point of the nostril contour and the nasopharynx outline was measured in mm parallel to the palatal plane. Descriptive statistics of the measurements were calculated for each age group. The lower limit of the 95% confidence intervals of the measurements was taken as a reference to design a swab shaft with marks that, at each age, delimitate a safety boundary for swab progression up to the posterior nasopharyngeal wall. The simplification of the procedure enabled by the newly designed nasopharyngeal swab is valuable to help healthcare providers perform specimen collection on children in a safe and effective way, perhaps under the less-than-ideal conditions possibly occurring in ‘point-of-need’ contexts.
Climate Change, Carbon Dioxide Emissions, and Medical Imaging Contribution
Authors: Picano Eugenio , Mangia Cristina , D’Andrea Antonello
Published: December 2022

Human activities have raised the atmosphere’s carbon dioxide (CO2) content by 50% in less than 200 years and by 10% in the last 15 years. Climate change is a great threat and presents a unique opportunity to protect cardiovascular health in the next decades. CO2 equivalent emission is the most convenient unit for measuring the greenhouse gas footprint corresponding to ecological cost. Medical imaging contributes significantly to the CO2 emissions responsible for climate change, yet current medical guidelines ignore the carbon cost. Among the common cardiac imaging techniques, CO2 emissions are lowest for transthoracic echocardiography (0.5–2 kg per exam), increase 10-fold for cardiac computed tomography angiography, and 100-fold for cardiac magnetic resonance. A conservative estimate of 10 billion medical examinations per year worldwide implies that medical imaging accounts for approximately 1% of the overall carbon footprint. In 2016, CO2 emissions from magnetic resonance imaging and computed tomography, calculated in 120 countries, accounted for 0.77% of global emissions. A significant portion of global greenhouse gas emissions is attributed to health care, which ranges from 4% in the United Kingdom to 10% in the United States. Assessment of carbon cost should be a part of the cost-benefit balance in medical imaging.