bluuebunny/crossref_metadata_2025 huggingface dataset:
“Stay at Home” during the COVID-19 Pandemic: Effects on Physical Activity and Sedentary Behavior in an Italian Academic Community
Authors: Mulasso Anna , Micheletti Cremasco Margherita , Cuomo Simone , Testa Andrea , Lattke Lynda Stella , De Lorenzo Aurelia , Rainoldi Alberto , Rabaglietti Emanuela
Published: January 2023

This study investigates changes in physical activity (PA) and sedentary behavior in an Italian university community during Phase 1 of SARS-CoV-2 lockdown (“stay at home” government decree, from March 8th to May 4th, 2020) compared to their habits prior to the COVID-19 pandemic. We also examine differences according to gender, university position, BMI categories, and sport participation. A total of 2596 people (median age 24, IQR 11 years; 70.8% women) filled out a survey after eight weeks of statutory confinement at home. The International Physical Activity Questionnaire measured PA and sedentary behavior in a typical week before and during lockdown. Physically inactive people passed from 10.9% to 35.0% before–during the lockdown. The total amount of PA decreased (median 2307 vs. 1367 MET-min/week; p < 0.001), while sedentary behavior increased (median 349 vs. 440 min/day; p < 0.001) between before and during the lockdown. The following categories showed a higher reduction in the total amount of PA: men when compared to women; people with normal weight when compared to pre-obese/obese people; and people who played sports when compared to those who did not play sports. There is a need to propose PA/exercise programs to counteract physical inactivity and sedentarism during a social emergency, with special attention to people who showed higher PA reduction.
First Data on PAE Levels in Surface Water in Lakes of the Eastern Coast of Baikal
Authors: Bazarsadueva Selmeg V. , Taraskin Vasilii V. , Budaeva Olga D. , Nikitina Elena P. , Zhigzhitzhapova Svetlana V. , Shiretorova Valentina G. , Bazarzhapov Tcogto Zh. , Radnaeva Larisa D.
Published: January 2023

The increasing consumption of phthalates (PAEs), along with their high toxicity and high mobility, poses a threat to the environment. This study presents initial data on the contents of six priority PAEs in the water of lakes located on the eastern shore of Lake Baikal-Arangatui, Bormashevoe, Dukhovoe, Kotokel, and Shchuchye. The mean total concentrations of the six PAEs in lakes Arangatui and Bormashevoe (low anthropogenic load) were comparable to those in Kotokel (medium anthropogenic load, 17.34 µg/L) but were significantly higher (p < 0.05) than in Dukhovoe and Shchuchye (high anthropogenic load, 10.49 and 2.30 µg/L, respectively). DBP and DEHP were the main PAEs in all samples. The DEHP content in lakes Arangatui and Bormashevoe was quite high, and at some sampling sites it exceeded the MACs established by Russian, U.S. EPA, and WHO regulations. The assessment showed that there is no potential risk to humans associated with the presence of PAEs in drinking water. However, the levels of DEHP, DBP, and DnOP in the water pose a potential threat to sensitive aquatic organisms, as shown by the calculated risk quotients (RQs). It is assumed that the origin of the phthalates in the studied lakes is both anthropogenic and biogenic.
Upconverting Nanoparticles as a New Bio-Imaging Strategy—Investigating Intracellular Trafficking of Endogenous Processes in Neural Tissue
Authors: Zajdel Karolina , Janowska Justyna , Frontczak-Baniewicz Małgorzata , Sypecka Joanna , Sikora Bożena
Published: January 2023

In recent years, rare-earth-doped upconverting nanoparticles (UCNPs) have been widely used in different life sciences due to their unique properties. Nanoparticles have become a multifunctional and promising new approach to neurobiological disorders and have shown extraordinary application potential to overcome the problems related to conventional treatment strategies. This study evaluated the internalization mechanisms, bio-distribution, and neurotoxicity of NaYF4:20%Yb3+,2%Er3+ UCNPs in rat organotypic hippocampal slices. TEM results showed that UCNPs were easily internalized by hippocampal cells and co-localized with selected organelles inside neurons and astrocytes. Moreover, the UCNPs were taken into the neurons via clathrin- and caveolae-mediated endocytosis. Propidium iodide staining and TEM analysis did not confirm the adverse effects of UCNPs on hippocampal slice viability and morphology. Therefore, UCNPs may be a potent tool for bio-imaging and testing new therapeutic strategies for brain diseases in the future.
Gene Therapy in ALS and SMA: Advances, Challenges and Perspectives
Authors: Lejman Jan , Panuciak Kinga , Nowicka Emilia , Mastalerczyk Angelika , Wojciechowska Katarzyna , Lejman Monika
Published: January 2023

Gene therapy is defined as the administration of genetic material to modify, manipulate gene expression or alter the properties of living cells for therapeutic purposes. Recent advances and improvements in this field have led to many breakthroughs in the treatment of various diseases. As a result, there has been an increasing interest in the use of these therapies to treat motor neuron diseases (MNDs), for which many potential molecular targets have been discovered. MNDs are neurodegenerative disorders that, in their most severe forms, can lead to respiratory failure and death, for instance, spinal muscular atrophy (SMA) or amyotrophic lateral sclerosis (ALS). Despite the fact that SMA has been known for many years, it is still one of the most common genetic diseases causing infant mortality. The introduction of drugs based on ASOs—nusinersen; small molecules—risdiplam; and replacement therapy (GRT)—Zolgensma has shown a significant improvement in both event-free survival and the quality of life of patients after using these therapies in the available trial results. Although there is still no drug that would effectively alleviate the course of the disease in ALS, the experience gained from SMA gene therapy gives hope for a positive outcome of the efforts to produce an effective and safe drug. The aim of this review is to present current progress and prospects for the use of gene therapy in the treatment of both SMA and ALS.
Adiponectin, Leptin, and Resistin Are Dysregulated in Patients Infected by SARS-CoV-2
Authors: Perrotta Fabio , Scialò Filippo , Mallardo Marta , Signoriello Giuseppe , D’Agnano Vito , Bianco Andrea , Daniele Aurora , Nigro Ersilia
Published: January 2023

Obesity, through adipose tissue (AT) inflammation and dysregulation, represents a critical factor for COVID-19; here, we investigated whether serum levels of adiponectin, HMW oligomers, leptin, and resistin are modulated and/or correlated with clinical and biochemical parameters of severe COVID-19 patients. This study included 62 severe COVID-19 patients; 62 age and sex-matched healthy subjects were recruited as a control group. Anthropometric and biochemical parameters were obtained and compared. Adiponectin, HMW oligomers, leptin, and resistin were analyzed by ELISA. The adiponectin oligomerization state was visualized by Western blotting. When compared to healthy subjects, total adiponectin levels were statistically lower in severe COVID-19 while, in contrast, the levels of leptin and resistin were statistically higher. Interestingly, HMW adiponectin oligomers negatively correlated with leptin and were positively associated with LUS scores. Resistin showed a positive association with IL-6, IL-2R, and KL-6. Our data strongly support that adipose tissue might play a functional role in COVID-19. Although it needs to be confirmed in larger cohorts, adiponectin HMW oligomers might represent a laboratory resource to predict patient seriousness. Whether adipokines can be integrated as a potential additional tool in the evolving landscape of biomarkers for the COVID-19 disease is still a matter of debate. Other studies are needed to understand the molecular mechanisms behind adipokine’s involvement in COVID-19.
CsCER6 and CsCER7 Influence Fruit Glossiness by Regulating Fruit Cuticular Wax Accumulation in Cucumber
Authors: Liu Xiaofeng , Ge Xinshuang , An Jingbo , Liu Xingwang , Ren Huazhong
Published: January 2023

Fruit glossiness is an important external fruit quality trait that greatly affects the marketability of fresh cucumber (Cucumis sativus) fruits. A few reports have suggested that the extent of cuticular wax loading influences the glossiness of the fruit surface. In the present study, we tested the wax contents of two inbred cucumber lines, comparing a line with waxy fruit (3401) and a line with glossy fruit (3413). Wax content analysis and dewaxing analysis demonstrate that fruit cuticular wax loads negatively correlate with fruit glossiness in cucumber. Identifying genes that were differentially expressed in fruit pericarps between 3401 and 3413 and genes induced by abscisic acid suggested that the wax biosynthesis gene CsCER6 (Cucumis sativus ECERIFERUM 6) and the regulatory gene CsCER7 may affect wax accumulation on cucumber fruit. Expression analysis via RT-qPCR, GUS-staining, and in situ hybridization revealed that CsCER6 and CsCER7 are abundantly expressed in the epidermis cells in cucumber fruits. Furthermore, the overexpression and RNAi lines of CsCER6 and CsCER7 showed dramatic effects on fruit cuticular wax contents, fruit glossiness, and cuticle permeability. Our results suggest that CsCER6 and CsCER7 positively regulate fruit cuticular wax accumulation and negatively influence fruit glossiness.
Layer-Wise Relevance Analysis for Motif Recognition in the Activation Pathway of the β2-Adrenergic GPCR Receptor
Authors: Gutiérrez-Mondragón Mario A. , König Caroline , Vellido Alfredo
Published: January 2023

G-protein-coupled receptors (GPCRs) are cell membrane proteins of relevance as therapeutic targets, and are associated to the development of treatments for illnesses such as diabetes, Alzheimer’s, or even cancer. Therefore, comprehending the underlying mechanisms of the receptor functional properties is of particular interest in pharmacoproteomics and in disease therapy at large. Their interaction with ligands elicits multiple molecular rearrangements all along their structure, inducing activation pathways that distinctly influence the cell response. In this work, we studied GPCR signaling pathways from molecular dynamics simulations as they provide rich information about the dynamic nature of the receptors. We focused on studying the molecular properties of the receptors using deep-learning-based methods. In particular, we designed and trained a one-dimensional convolution neural network and illustrated its use in a classification of conformational states: active, intermediate, or inactive, of the β2-adrenergic receptor when bound to the full agonist BI-167107. Through a novel explainability-oriented investigation of the prediction results, we were able to identify and assess the contribution of individual motifs (residues) influencing a particular activation pathway. Consequently, we contribute a methodology that assists in the elucidation of the underlying mechanisms of receptor activation–deactivation.
Development of a Method for the In Vivo Generation of Allogeneic Hearts in Chimeric Mouse Embryos
Authors: Founta Konstantina-Maria , Tourkodimitri Magdalini-Ioanna , Kanaki Zoi , Bisti Sylvia , Papanayotou Costis
Published: January 2023

Worldwide, there is a great gap between the demand and supply of organs for transplantations. Organs generated from the patients’ cells would not only solve the problem of transplant availability but also overcome the complication of incompatibility and tissue rejection by the host immune system. One of the most promising methods tested for the production of organs in vivo is blastocyst complementation (BC). Regrettably, BC is not suitable for the creation of hearts. We have developed a novel method, induced blastocyst complementation (iBC), to surpass this shortcoming. By applying iBC, we generated chimeric mouse embryos, made up of “host” and “donor” cells. We used a specific cardiac enhancer to drive the expression of the diphtheria toxin gene (dtA) in the “host” cells, so that these cells are depleted from the developing hearts, which now consist of “donor” cells. This is a proof-of-concept study, showing that it is possible to produce allogeneic and ultimately, xenogeneic hearts in chimeric organisms. The ultimate goal is to generate, in the future, human hearts in big animals such as pigs, from the patients’ cells, for transplantations. Such a system would generate transplants in a relatively short amount of time, improving the quality of life for countless patients around the world.
The Role of Neutrophils in Lower Limb Peripheral Artery Disease: State of the Art and Future Perspectives
Authors: Buso Giacomo , Faggin Elisabetta , Rosenblatt-Velin Nathalie , Pellegrin Maxime , Galliazzo Silvia , Calanca Luca , Rattazzi Marcello , Mazzolai Lucia
Published: January 2023

In recent years, increasing attention has been paid to the role of neutrophils in cardiovascular (CV) disease (CVD) with evidence supporting their role in the initiation, progression, and rupture of atherosclerotic plaque. Although these cells have long been considered as terminally differentiated cells with a relatively limited spectrum of action, recent research has revealed intriguing novel cellular functions, including neutrophil extracellular trap (NET) generation and inflammasome activation, which have been linked to several human diseases, including CVD. While most research to date has focused on the role of neutrophils in coronary artery and cerebrovascular diseases, much less information is available on lower limb peripheral artery disease (PAD). PAD is a widespread condition associated with great morbidity and mortality, though physician and patient awareness of the disease remains low. To date, several studies have produced some evidence on the role of certain biomarkers of neutrophil activation in this clinical setting. However, the etiopathogenetic role of neutrophils, and in particular of some of the newly discovered mechanisms, has yet to be fully elucidated. In the future, complementary assessment of neutrophil activity should improve CV risk stratification and provide personalized treatments to patients with PAD. This review aims to summarize the basic principles and recent advances in the understanding of neutrophil biology, current knowledge about the role of neutrophils in atherosclerosis, as well as available evidence on their role of PAD.
Plasma Concentrations of Neurofilament Light Chain Protein and Brain-Derived Neurotrophic Factor as Consistent Biomarkers of Cognitive Impairment in Alcohol Use Disorder
Authors: Requena-Ocaña Nerea , Araos Pedro , Serrano-Castro Pedro J. , Flores-López María , García-Marchena Nuria , Oliver-Martos Begoña , Ruiz Juan Jesús , Gavito Ana , Pavón Francisco Javier , Serrano Antonia , Mayoral Fermín , Suarez Juan , Fonseca Fernando Rodríguez de
Published: January 2023

For a long time, Substance Use Disorders (SUDs) were not considered a component in the etiology of dementia. The fifth edition of the Diagnostic and Statistical Manual of Mental Disorders introduced substance-induced neurocognitive disorders, incorporating this notion to clinical practice. However, detection and monitoring of neurodegenerative processes in SUD patients remain a major clinical challenge, especially when early diagnosis is required. In the present study, we aimed to investigate new potential biomarkers of neurodegeneration that could predict cognitive impairment in SUD patients: the circulating concentrations of Neurofilament Light chain protein (NfL) and Brain-Derived Neurotrophic Factor (BDNF). Sixty SUD patients were compared with twenty-seven dementia patients and forty healthy controls. SUD patients were recruited and assessed using the Psychiatric Research Interview for Substance and Mental (PRISM) and a battery of neuropsychological tests, including the Montreal Cognitive Assessment test for evaluation of cognitive impairment. When compared to healthy control subjects, SUD patients showed increases in plasma NfL concentrations and NfL/BDNF ratio, as well as reduced plasma BDNF levels. These changes were remarkable in SUD patients with moderate–severe cognitive impairment, being comparable to those observed in dementia patients. NfL concentrations correlated with executive function and memory cognition in SUD patients. The parameters “age”, “NfL/BDNF ratio”, “first time alcohol use”, “age of onset of alcohol use disorder”, and “length of alcohol use disorder diagnosis” were able to stratify our SUD sample into patients with cognitive impairment from those without cognitive dysfunction with great specificity and sensibility. In conclusion, we propose the combined use of NfL and BDNF (NfL/BDNF ratio) to monitor substance-induced neurocognitive disorder.
Inhibition of ANO1 by Cis- and Trans-Resveratrol and Their Anticancer Activity in Human Prostate Cancer PC-3 Cells
Authors: Jeon Dongkyu , Jo Minjae , Lee Yechan , Park So-Hyeon , Phan Hong Thi Lam , Nam Joo Hyun , Namkung Wan
Published: January 2023

Anoctamin1 (ANO1), a calcium-activated chloride channel, is involved in the proliferation, migration, and invasion of various cancer cells including head and neck squamous cell carcinoma, lung cancer, and prostate cancer. Inhibition of ANO1 activity or downregulation of ANO1 expression in these cancer cells is known to exhibit anticancer effects. Resveratrol, a natural polyphenol abundant in wines, grapes, berries, soybeans, and peanuts, shows a wide variety of biological effects including anti-inflammatory, antioxidant, and anticancer activities. In this study, we investigated the effects of two stereoisomers of resveratrol on ANO1 activity and found that cis- and trans-resveratrol inhibited ANO1 activity with different potencies. Cis- and trans-resveratrol inhibited ANO1 channel activity with IC50 values of 10.6 and 102 μM, respectively, and had no significant effect on intracellular calcium signaling at 10 and 100 μM, respectively. In addition, cis-resveratrol downregulated mRNA and protein expression levels of ANO1 more potently than trans-resveratrol in PC-3 prostate cancer cells. Cis- and trans-resveratrol significantly reduced cell proliferation and cell migration in an ANO1-dependent manner, and both resveratrol isomers strongly increased caspase-3 activity, PARP cleavage, and apoptotic sub-G1 phase ratio in PC-3 cells. These results revealed that cis-resveratrol is a potent inhibitor of ANO1 and exhibits ANO1-dependent anticancer activity against human metastatic prostate cancer PC-3 cells.
Proton-Pump Inhibitors Suppress T Cell Response by Shifting Intracellular Zinc Distribution
Authors: Liu Wenlei , Jakobs Jana , Rink Lothar
Published: January 2023

Proton-pump inhibitors (PPI), e.g., omeprazole or pantoprazole, are the most widely used drugs for various gastrointestinal diseases. However, more and more side effects, especially an increased risk of infections, have been reported in recent years. The underlying mechanism has still not yet been fully uncovered. Hence, in this study, we analyzed the T cell response after treatment with pantoprazole in vitro. Pantoprazole preincubation reduced the production and secretion of interferon (IFN)-γ and interleukin (IL)-2 after the T cells were activated with phytohemagglutinin (PHA)-L or toxic shock syndrome toxin-1 (TSST-1). Moreover, a lower zinc concentration in the cytoplasm and a higher concentration in the lysosomes were observed in the pantoprazole-treated group compared to the untreated group. We also tested the expression of the zinc transporter Zrt- and Irt-like protein (Zip)8, which is located in the lysosomal membrane and plays a key role in regulating intracellular zinc distribution after T cell activation. Pantoprazole reduced the expression of Zip8. Furthermore, we measured the expression of cAMP-responsive element modulator (CREM) α, which directly suppresses the expression of IL-2, and the expression of the phosphorylated cAMP response element-binding protein (pCREB), which can promote the expression of IFN-γ. The expression of CREMα was dramatically increased, and different isoforms appeared, whereas the expression of pCREB was downregulated after the T cells were treated with pantoprazole. In conclusion, pantoprazole downregulates IFN-γ and IL-2 expression by regulating the expression of Zip8 and pCREB or CREMα, respectively.
Subacute Ruminal Acidosis as a Potential Factor that Induces Endometrium Injury in Sheep
Authors: Zeng Jianlin , Lv Jianshu , Duan Hongwei , Yang Shuai , Wu Jianxin , Yan Zhenxing , Zhang Rong , Hu Junjie , Zhang Yong
Published: January 2023

The demand for economic benefits has led to an increase in the proportion of high-concentrate (HC) feed in the ruminant diet, resulting in an increased incidence of subacute ruminal acidosis (SARA). During SARA, a high concentration of lipopolysaccharide (LPS) translocated in the rumen induces a systemic inflammatory response. Inflammatory diseases, such as endometritis and mastitis, are often associated with SARA; however, in sheep, the mechanism of the effect of SARA on the endometrium has rarely been reported. Therefore, the aim of this study was to investigate, for the first time, the influence of LPS translocation on endometrial tight junctions (TJs) during SARA in sheep. The results showed that LPS and TNFα levels in the ruminal fluid, serum, and endometrial tissue supernatant during SARA increased, transcription levels of TLR4, NFκB, and TNFα in the endometrium increased, the protein expression level of claudin-1 in the endometrium increased, and the protein expression level of occludin decreased. 17β-estradiol (E2) inhibits claudin-1 protein expression and promotes occludin expression, and progesterone (P4) promotes claudin-1 protein expression and inhibits occludin protein expression. E2 and P4 regulate claudin-1 and occludin protein expression through their receptor pathways. Here, we found that LPS hindered the regulatory effect of E2 and P4 on endometrial TJs by inhibiting their receptor expression. The results of this study indicate that HC feeding can cause SARA-induced LPS translocation in sheep, increase susceptibility to systemic inflammation, induce the endometrial inflammatory response, and cause endometrial epithelial TJ damage directly and/or by obstructing E2 and P4 function. LPS translocation caused by SARA has also been suggested to induce an endometrial inflammatory response, resulting in endometrial epithelial barrier damage and physiological dysfunction, which seriously affects ruminant production. Therefore, this study provides new evidence that SARA is a potential factor that induces systemic inflammation in ruminants. It provides theoretical support for research on the prevention of endometritis in ruminants.
Effects and Mechanisms Activated by Treatment with Cationic, Anionic and Zwitterionic Liposomes on an In Vitro Model of Porcine Pre-Pubertal Sertoli Cells
Authors: Collodel Giulia , Moretti Elena , Noto Daria , Corsaro Roberta , Signorini Cinzia , Bonechi Claudia , Cangeloni Lorenzo , Luca Giovanni , Arato Iva , Mancuso Francesca
Published: January 2023

Liposomes have been successfully used as drug-delivery vehicles, but there are no clinical studies on improved fertility and the few reported experimental studies have been performed in animal models far from humans. The aim of this paper was to study the effects of treatment with cationic, anionic and zwitterionic liposomes on our superior mammalian model of porcine prepubertal Sertoli cells (SCs) to find a carrier of in vitro test drugs for SCs. Porcine pre-pubertal SCs cultures were incubated with different liposomes. Viability, apoptosis/necrosis status (Annexin-V/Propidium iodide assay), immunolocalisation of β-actin, vimentin, the phosphorylated form of AMP-activated protein Kinase (AMPK)α and cell ultrastructure (Transmission Electron Microscopy, TEM) were analysed. Zwitterionic liposomes did not determine changes in the cell cytoplasm. The incubation with anionic and cationic liposomes modified the distribution of actin and vimentin filaments and increased the levels of the phosphorylated form of AMPKα. The Annexin/Propidium Iodide assay suggested an increase in apoptosis. TEM analysis highlighted a cytoplasmic vacuolisation. In conclusion, these preliminary data indicated that zwitterionic liposomes were the best carrier to use in an in vitro study of SCs to understand the effects of molecules or drugs that could have a clinical application in the treatment of certain forms of male infertility.
Kalkitoxin: A Potent Suppressor of Distant Breast Cancer Metastasis
Authors: Shrestha Saroj Kumar , Min Kyung Hyun , Kim Se Woong , Kim Hyoungsu , Gerwick William H. , Soh Yunjo
Published: January 2023

Bone metastasis resulting from advanced breast cancer causes osteolysis and increases mortality in patients. Kalkitoxin (KT), a lipopeptide toxin derived from the marine cyanobacterium Moorena producens (previously Lyngbya majuscula), has an anti-metastatic effect on cancer cells. We verified that KT suppressed cancer cell migration and invasion in vitro and in animal models in the present study. We confirmed that KT suppressed osteoclast-soup-derived MDA-MB-231 cell invasion in vitro and induced osteolysis in a mouse model, possibly enhancing/inhibiting metastasis markers. Furthermore, KT inhibits CXCL5 and CXCR2 expression, suppressing the secondary growth of breast cancer cells on the bone, brain, and lungs. The breast-cancer-induced osteolysis in the mouse model further reveals that KT plays a protective role, judging by micro-computed tomography and immunohistochemistry. We report for the first time the novel suppressive effects of KT on cancer cell migration and invasion in vitro and on MDA-MB-231-induced bone loss in vivo. These results suggest that KT may be a potential therapeutic drug for the treatment of breast cancer metastasis.
DMSO-Induced Unfolding of the Antifungal Disulfide Protein PAF and Its Inactive Variant: A Combined NMR and DSC Study
Authors: Czajlik András , Batta Ágnes , Kerner Kinga , Fizil Ádám , Hajdu Dorottya , Raics Mária , Kövér Katalin E. , Batta Gyula
Published: January 2023

PAF and related antifungal proteins are promising antimicrobial agents. They have highly stable folds around room temperature due to the presence of 3–4 disulfide bonds. However, unfolded states persist and contribute to the thermal equilibrium in aqueous solution, and low-populated states might influence their biological impact. To explore such equilibria during dimethyl sulfoxide (DMSO)-induced chemical unfolding, we studied PAF and its inactive variant PAFD19S using nuclear magnetic resonance (NMR) and differential scanning calorimetry (DSC). According to the NMR monitoring at 310 K, the folded structures disappear above 80 v/v% DMSO concentration, while the unfolding is completely reversible. Evaluation of a few resolved peaks from viscosity-compensated 15N-1H HSQC spectra of PAF yielded ∆G = 23 ± 7 kJ/M as the average value for NMR unfolding enthalpy. The NMR-based structures of PAF and the mutant in 50 v/v% DMSO/H2O mixtures were more similar in the mixed solvents then they were in water. The 15N NMR relaxation dynamics in the same mixtures verified the rigid backbones of the NMR-visible fractions of the proteins; still, enhanced dynamics around the termini and some loops were observed. DSC monitoring of the Tm melting point showed parabolic dependence on the DMSO molar fraction and suggested that PAF is more stable than the inactive PAFD19S. The DSC experiments were irreversible due to the applied broad temperature range, but still suggestive of the endothermic unfolding of PAF.
Infection and Persistence of Coxiella burnetii Clinical Isolate in the Placental Environment
Authors: Zarza Sandra Madariaga , Militello Muriel , Gay Laetitia , Levasseur Anthony , Lepidi Hubert , Bechah Yassina , Mezouar Soraya , Mege Jean-Louis
Published: January 2023

Infection by Coxiella burnetii, the etiological agent of Q fever, poses the risk of causing severe obstetrical complications in pregnant women. C. burnetii is known for its placental tropism based on animal models of infection. The Nine Mile strain has been mostly used to study C. burnetii pathogenicity but the contribution of human isolates to C. burnetii pathogenicity is poorly understood. In this study, we compared five C. burnetii isolates from human placentas with C. burnetii strains including Nine Mile (NM) as reference. Comparative genomic analysis revealed that the Cb122 isolate was distinct from other placental isolates and the C. burnetii NM strain with a set of unique genes involved in energy generation and a type 1 secretion system. The infection of Balb/C mice with the Cb122 isolate showed higher virulence than that of NM or other placental isolates. We evaluated the pathogenicity of the Cb122 isolate by in vitro and ex vivo experiments. As C. burnetii is known to infect and survive within macrophages, we isolated monocytes and placental macrophages from healthy donors and infected them with the Cb122 isolate and the reference strain. We showed that bacteria from the Cb122 isolate were less internalized by monocyte-derived macrophages (MDM) than NM bacteria but the reference strain and the Cb122 isolate were similarly internalized by placental macrophages. The Cb122 isolate and the reference strain survived similarly in the two macrophage types. While the Cb122 isolate and the NM strain stimulated a poorly inflammatory program in MDM, they elicited an inflammatory program in placenta macrophages. We also reported that the Cb122 isolate and NM strain were internalized by trophoblastic cell lines and primary trophoblasts without specific replicative profiles. Placental explants were then infected with the Cb122 isolate and the NM strain. The bacteria from the Cb122 isolate were enriched in the chorionic villous foetal side. It is likely that the Cb122 isolate exhibited increased virulence in the multicellular environment provided by explants. Taken together, these results showed that the placental isolate of C. burnetii exhibits a specific infectious profile but its pathogenic role is not as high as the host immune response in pregnant women.
Effect of Octacalcium Phosphate Crystals on the Osteogenic Differentiation of Tendon Stem/Progenitor Cells In Vitro
Authors: Liu Xianchen , Shiwaku Yukari , Hamai Ryo , Tsuchiya Kaori , Takahashi Tetsu , Suzuki Osamu
Published: January 2023

Synthetic octacalcium phosphate (OCP) activates bone tissue-related cells, such as osteoblasts, osteoclasts, and vascular endothelial cells. However, the effect of OCP on tendon-related cell activation remains unknown. This study examined the response of rat tendon stem/progenitor cells (TSPCs) to OCP and related calcium phosphate crystals in vitro. TSPCs were cultured with OCP and Ca-deficient hydroxyapatite (CDHA) obtained from the original OCP hydrolysis to assess the activity of alkaline phosphatase (ALP) and the expression of osteogenesis-related genes. Compared with CDHA, the effect of OCP on promoting the osteogenic differentiation of TSPCs was apparent: the ALP activity and mRNA expression of RUNX2, Col1a1, OCN, and OPN were higher in OCP than in CDHA. To estimate the changes in the chemical environment caused by OCP and CDHA, we measured the calcium ion (Ca2+) and inorganic phosphate (Pi) ion concentrations and pH values of the TSPCs medium. The results suggest that the difference in the osteogenic differentiation of the TSPCs is related to the ionic environment induced by OCP and CDHA, which could be related to the progress of OCP hydrolysis into CDHA. These results support the previous in vivo observation that OCP has the healing function of rabbit rotator cuff tendon in vivo.
Kaposi’s Sarcoma-Associated Herpesvirus ORF21 Enhances the Phosphorylation of MEK and the Infectivity of Progeny Virus
Authors: Yamaguchi Tatsuo , Watanabe Tadashi , Iwaisako Yuki , Fujimuro Masahiro
Published: January 2023

Kaposi’s sarcoma-associated herpesvirus (KSHV), also known as human herpesvirus-8, is the causative agent of Kaposi’s sarcoma, Castleman’s disease, and primary effusion lymphoma. Although the functions of the viral thymidine kinases (vTK) of herpes simplex virus-1/2 are well understood, that of KSHV ORF21 (an ortholog of vTK) is largely unknown. Here, we investigated the role of ORF21 in lytic replication and infection by generating two ORF21-mutated KSHV BAC clones: ORF21-kinase activity deficient KSHV (21KD) and stop codon-induced ORF21-deleted KSHV (21del). The results showed that both ORF21 mutations did not affect viral genome replication, lytic gene transcription, or the production of viral genome-encapsidated particles. The ORF21 molecule-dependent function, other than the kinase function of ORF21, was involved in the infectivity of the progeny virus. ORF21 was expressed 36 h after the induction of lytic replication, and endogenously expressed ORF21 was localized in the whole cytoplasm. Moreover, ORF21 upregulated the MEK phosphorylation and anchorage-independent cell growth. The inhibition of MEK signaling by U0126 in recipient target cells suppressed the number of progeny virus-infected cells. These suggest that ORF21 transmitted as a tegument protein in the progeny virus enhances the new infection through MEK up-regulation in the recipient cell. Our findings indicate that ORF21 plays key roles in the infection of KSHV through the manipulation of the cellular function.
Fabrication and Performance Evaluation of Gelatin/Sodium Alginate Hydrogel-Based Macrophage and MSC Cell-Encapsulated Paracrine System with Potential Application in Wound Healing
Authors: Yao Hang , Yuan Xiaohui , Wu Zhonglian , Park Sumin , Zhang Wang , Chong Hui , Lin Liwei , Piao Yuanzhe
Published: January 2023

A gelatin/sodium alginate-based hydrogel microsphere has been fabricated after reaction condition optimization. Macrophages (RAW246.7) and adipose mesenchymal stem cells (ADSC) have been subsequently encapsulated in the microsphere in order to construct a 3D paracrine system for wound healing treatment. The synthesized microsphere displayed neglectable cytotoxicity toward both encapsulated cells until 10 days of incubation, indicating promising biocompatibility of the microsphere. A qRT-PCR and ELISA experiment revealed positive regulation of cytokines (Arg-1, IL-6, IL-8, IL-10, bFGF, HGF, VEGF, TLR-1, and CXCL13) expression regarding macrophage phenotype transformation and anti-inflammatory performance both inside the microsphere and in the microenvironment of established in vitro inflammatory model. Additionally, positive tendency of cytokine expression benefit wound healing was more pronounced in a fabricated 3D paracrine system than that of a 2D paracrine system. Furthermore, the 3D paracrine system exhibited more efficiently in the wound healing rate compared to the 2D paracrine system in an in vitro model. These results suggested the current paracrine system could be potentially used as a robust wound healing dressing.
Metal Complexes as Promising Matrix Metalloproteinases Regulators
Authors: Nguyen Yen Thi , Kim Namdoo , Lee Hyuck Jin
Published: January 2023

Nowadays, cancers and dementia, such as Alzheimer’s disease, are the most fatal causes of death. Many studies tried to understand the pathogenesis of those diseases clearly and develop a promising way to treat the diseases. Matrix metalloproteinases (MMPs) have been reported to be involved in the pathology of cancers and AD through tumor cell movement and amyloid degradation. Therefore, control of the levels and actions of MMPs, especially MMP-2 and MMP-9, is necessary to care for and/or cure cancer and AD. Various molecules have been examined for their potential application as regulators of MMPs expression and activity. Among the molecules, multiple metal complexes have shown advantages, including simple synthesis, less toxicity and specificity toward MMPs in cancer cells or in the brain. In this review, we summarize the recent studies and knowledge of metal complexes (e.g., Pt-, Ru-, Au-, Fe-, Cu-, Ni-, Zn-, and Sn-complexes) targeting MMPs and their potentials for treating and/or caring the most fatal human diseases, cancers and AD.
Molecular Mechanism of Cold Tolerance of Centipedegrass Based on the Transcriptome
Authors: Liu Yingjie , Xiong Yi , Zhao Junming , Bai Shiqie , Li Daxu , Chen Limin , Feng Junjie , Li Yingzhu , Ma Xiao , Zhang Jianbo
Published: January 2023

Low temperature is an important limiting factor in the environment that affects the distribution, growth and development of warm-season grasses. Transcriptome sequencing has been widely used to mine candidate genes under low-temperature stress and other abiotic stresses. However, the molecular mechanism of centipedegrass in response to low-temperature stress was rarely reported. To understand the molecular mechanism of centipedegrass in response to low-temperature stress, we measured physiological indicators and sequenced the transcriptome of centipedegrass under different stress durations. Under cold stress, the SS content and APX activity of centipedegrass increased while the SOD activity decreased; the CAT activity, POD activity and flavonoid content first increased and then decreased; and the GSH-Px activity first decreased and then increased. Using full-length transcriptome and second-generation sequencing, we obtained 38.76 G subreads. These reads were integrated into 177,178 isoforms, and 885 differentially expressed transcripts were obtained. The expression of AUX_IAA and WRKY transcription factors and HSF transcription-influencing factors increased during cold stress. Through KEGG enrichment analysis, we determined that arginine and proline metabolism, plant circadian rhythm, plant hormone signal transduction and the flavonoid biosynthesis pathways played important roles in the cold stress resistance of centipedegrass. In addition, by using weighted gene coexpression network analysis (WGCNA), we determined that the turquoise module was significantly correlated with SS content and APX activity, while the blue module was significantly negatively correlated with POD and CAT activity. This paper is the first to report the response of centipedegrass to cold stress at the transcriptome level. Our results help to clarify the molecular mechanisms underlying the cold tolerance of warm-season grasses.
Interleukin-19 Levels Are Increased in Palmoplantar Pustulosis and Reduced following Apremilast Treatment
Authors: Wolk Kerstin , Wilsmann-Theis Dagmar , Witte Katrin , Brembach Theresa-Charlotte , Kromer Christian , Gerdes Sascha , Ghoreschi Kamran , Reich Kristian , Mössner Rotraut , Sabat Robert
Published: January 2023

Palmoplantar pustulosis (PPP) is a chronic inflammatory skin disease characterised by neutrophilic granulocyte (neutrophil)-filled pustules on the palms and soles. The pathogenesis of PPP is poorly understood. This study conducted an identification of the immune mediators associated with PPP and an exploration of apremilast treatment effects on them. We screened for immune mediators elevated in blood taken from 68 patients with PPP versus control participants and included the most promising parameters in the protocol of phase the 2, multicentre study of apremilast (PDE4 inhibitor) in 21 patients with moderate-to-severe PPP (APLANTUS; EudraCT 2016-005122-11) for respective analysis of blood and skin samples of study patients. We investigated stimulated neutrophils and three-dimensional reconstituted epidermis cultures. Interleukin (IL)-19 was found to be the most upregulated immune mediator in the blood of PPP patients. IL-19 serum levels were independent of patients’ age, gender, and BMI but were associated with strongly upregulated cutaneous IL-19 expression and correlated with the number of palmoplantar pustules. In patients participating in the APLANTUS study, apremilast reduced pustules more effectively than erythema and scaling. Moreover, this treatment significantly reduced IL-19 blood and skin levels. The reduction in IL-19 blood levels at week 4 correlated with the reduction in pustule counts at week 20 (end of treatment). IL-19 was expressed by neutrophils activated in vitro and induced CXCL6, a neutrophil-attracting chemokine, in epidermis models. This work demonstrates elevated IL-19 levels in the blood and skin of PPP patients and suggests a relevant role of this cytokine in the appearance of pustules in this disorder. It also suggests the suitability of IL-19 blood levels as a predictive biomarker for the treatment response of PPP patients, which should be validated in further studies.
Influence of the Method of Fe Deposition on the Surface of Hydrolytic Lignin on the Activity in the Process of Its Conversion in the Presence of CO2
Authors: Medvedev Artem A. , Kustov Alexander L. , Beldova Daria A. , Kalmykov Konstantin B. , Mashkin Mikhail Yu. , Shesterkina Anastasia A. , Dunaev Sergey F. , Kustov Leonid M.
Published: January 2023

Hydrolytic lignin is one of the non-demanded carbon materials. Its CO2-assisted conversion is an important way to utilize it. The use of the catalysts prepared by metal deposition on the surface of hydrolytic lignin makes it possible to apply milder conditions of the conversion process with CO2 and to improve the economic indicators. The development of methods of deposition of the active phase is a problem of high importance for any heterogeneous catalytic processes. This work aimed at investigating the influence of the conditions of iron deposition on the surface of hydrolytic lignin on the process of CO2-assisted conversion of lignin. Different Fe precursors (Fe(NO3)3, FeSO4, Fe2(SO4)3), solvents (water, isopropanol, acetone, and ethanol), and concentrations of the solution were used; the properties of Fe/lignin composites were estimated by SEM, EDX, TEM, XRD methods and catalytic tests. All the prepared samples demonstrate a higher conversion compared to starting lignin itself in the carbon dioxide-assisted conversion process. The carbon dioxide conversion was up to 66% at 800 °C for the sample prepared from Fe(NO3)3 using a twofold water volume compared to incipient wetness water volume as a solvent (vs. 39% for pure lignin).
Calcium-Signalling in Human Glaucoma Lamina Cribrosa Myofibroblasts
Authors: Irnaten Mustapha , O’Brien Colm J.
Published: January 2023

Glaucoma is one of the most common causes of treatable visual impairment in the developed world, affecting approximately 64 million people worldwide, some of whom will be bilaterally blind from irreversible optic nerve damage. The optic nerve head is a key site of damage in glaucoma where there is fibrosis of the connective tissue in the lamina cribrosa (LC) extracellular matrix. As a ubiquitous second messenger, calcium (Ca2+) can interact with various cellular proteins to regulate multiple physiological processes and contribute to a wide range of diseases, including cancer, fibrosis, and glaucoma. Our research has shown evidence of oxidative stress, mitochondrial dysfunction, an elevated expression of Ca2+ entry channels, Ca2+-dependent pumps and exchangers, and an abnormal rise in cytosolic Ca2+ in human glaucomatous LC fibroblast cells. We have evidence that this increase is dependent on Ca2+ entry channels located in the plasma membrane, and its release is from internal stores in the endoplasmic reticulum (ER), as well as from the mitochondria. Here, we summarize some of the molecular Ca2+-dependent mechanisms related to this abnormal Ca2+-signalling in human glaucoma LC cells, with a view toward identifying potential therapeutic targets for ongoing optic neuropathy.
CXADR: From an Essential Structural Component to a Vital Signaling Mediator in Spermatogenesis
Authors: Zhang Yang , Lui Wing-Yee
Published: January 2023

Canonical coxsackievirus and adenovirus receptor (CXADR) is a transmembrane component of cell junctions that is crucial for cardiac and testicular functions via its homophilic and heterophilic interaction. CXADR is expressed in both Sertoli cells and germ cells and is localized mainly at the interface between Sertoli-Sertoli cells and Sertoli-germ cells. Knockout of CXADR in mouse Sertoli cells specifically impairs male reproductive functions, including a compromised blood-testis barrier, apoptosis of germ cells, and premature loss of spermatids. Apart from serving as an important component for cell junctions, recent progress has showed the potential roles of CXADR as a signaling mediator in spermatogenesis. This review summarizes current research progress related to the regulation and role of CXADR in spermatogenesis as well as in pathological conditions. We hope this review provides some future directions and a blueprint to promote the further study on the roles of CXADR.
Evaluation of Chitosans as Coagulants—Flocculants to Improve Sand Filtration for Drinking Water Treatment
Authors: Holmes Eleanor B. , Oza Hemali H. , Bailey Emily S. , Sobsey Mark D.
Published: January 2023

The World Health Organization (WHO) reports that two billion people worldwide lack access to safely managed water sources, including 1.2 billion who already have access to improved water sources. In many countries, household point-of-use (POU) water-treatment options are used to remove or deactivate microorganisms in water, but not all POU technologies meet WHO performance requirements to achieve safe drinking water. To improve the effectiveness of POU technologies, the use of multiple treatment barriers should be used as a way to increase overall treatment performance. The focus of this research is to evaluate multiple barrier treatment using chitosan, an organic coagulant–flocculant, to improve microbial and turbidity reductions in combination with sand filtration. Bench-scale intermittently operated sand filters with 16 cm layers of sands of two different grain sizes representing slow and rapid sand filters were dosed daily over 57 days with microbially spiked surface water volumes corresponding to household use. E. coli bacteria and MS2 coliphage virus reductions were quantified biweekly (N = 17) using culture methods. Bacteria and virus removals were significantly improved over sand filtration without chitosan pretreatment (Wilcoxon Rank-Sum, p < 0.05). When water was pretreated at an optimal chitosan dose of 10 mg/L followed by sand filtration, log10 reductions in bacteria and viruses met the two-star WHO performance level of effectiveness. Microbial and turbidity reductions generally improved over the filter operating period but showed no trends with filtration rates.
Biomimetic Nanopillar Silicon Surfaces Rupture Fungal Spores
Authors: Linklater Denver P. , Le Phuc H. , Aburto-Medina Arturo , Crawford Russell J. , Maclaughlin Shane , Juodkazis Saulius , Ivanova Elena P.
Published: January 2023

The mechano-bactericidal action of nanostructured surfaces is well-documented; however, synthetic nanostructured surfaces have not yet been explored for their antifungal properties toward filamentous fungal species. In this study, we developed a biomimetic nanostructured surface inspired by dragonfly wings. A high-aspect-ratio nanopillar topography was created on silicon (nano-Si) surfaces using inductively coupled plasma reactive ion etching (ICP RIE). To mimic the superhydrophobic nature of insect wings, the nano-Si was further functionalised with trichloro(1H,1H,2H,2H-perfluorooctyl)silane (PFTS). The viability of Aspergillus brasiliensis spores, in contact with either hydrophobic or hydrophilic nano-Si surfaces, was determined using a combination of standard microbiological assays, confocal laser scanning microscopy (CLSM), and focused ion beam scanning electron microscopy (FIB-SEM). Results indicated the breakdown of the fungal spore membrane upon contact with the hydrophilic nano-Si surfaces. By contrast, hydrophobised nano-Si surfaces prevented the initial attachment of the fungal conidia. Hydrophilic nano-Si surfaces exhibited both antifungal and fungicidal properties toward attached A. brasisiensis spores via a 4-fold reduction of attached spores and approximately 9-fold reduction of viable conidia from initial solution after 24 h compared to their planar Si counterparts. Thus, we reveal, for the first time, the physical rupturing of attaching fungal spores by biomimetic hydrophilic nanostructured surfaces.