bluuebunny/crossref_metadata_2025 huggingface dataset:
Record-Breaking Slow Temperature Evolution of Spring Water During 2020 and Its Impacts on Spring Bloom in the Yellow Sea
Authors: Kim Go-Un , Lee Keunjong , Lee Jaeik , Jeong Jin-Yong , Lee Meehye , Jang Chan Joo , Ha Kyung-Ja , Nam SungHyun , Noh Jae Hoon , Kim Yong Sun
Published: April 2022

Unprecedented retardation of spring water temperature rising during the 2020 pandemic year was identified in the Socheongcho Ocean Research Station within the northeastern basin of the Yellow Sea: an exceptionally high water temperature anomaly in March and a contrasted low-temperature anomaly in May. The slowest temperature evolution was principally caused by the significant increase in latent heat releases in April (117% higher than the climatology of 1982–2019). Strong northwesterly winds generated these exceptional heat fluxes associated with the dipole-like atmospheric circulation pattern over Siberia and the East Sea (Japan Sea). Besides, warm winter water facilitated the enhanced release of latent heat fluxes as a precondition. The oceanic heat redistribution partially supported the cold anomaly in the surface layer up to the middle of May through positive feedback between the low surface temperature and the active entrainment associated with tidal turbulent mixing. The resultant low temperature at the surface weakened the vertical stratification, both impeding the activation of phytoplankton’s photosynthesis albeit under the eutrophic surface layer, consequently resulting in the delayed and suppressed spring bloom during 2020. Since such extreme events are anticipated to occur more frequently under global warming, our results highlight the importance of continuously monitoring multi-disciplinary environmental conditions, climate extremes, and their impact on the Yellow Sea marine ecosystem.
On the Utilization of Dietary Glycerol in Carnivorous Fish - Part I: Insights Into Hepatic Carbohydrate Metabolism of Juvenile Rainbow Trout (Oncorhynchus mykiss) and European Seabass (Dicentrarchus labrax)
Authors: Viegas Ivan , Tavares Ludgero C. , Plagnes-Juan Elisabeth , Silva Emanuel , Rito João , Marandel Lucie , Palma Mariana , Ozório Rodrigo O. A. , Magnoni Leonardo J. , Panserat Stéphane
Published: April 2022

Glycerol is a 3-carbon sugar alcohol successfully employed as an alternative feed ingredient for land-farmed animals and more recently for farmed fish. While most studies address zootechnical performance, few have delved into the metabolic utilization of dietary glycerol. A growth trial was performed using diets with increasing levels of glycerol (0, 2.5 and 5%) on two relevant species for aquaculture: rainbow trout (8-week trial; 3 tank per diet/25 fish per tank, on a 15 ± 1°C flow-through freshwater system); and European seabass (6-week trial; 5 tank per diet/6-8 fish per tank on a 21°C indoor saltwater RAS system). After this period, fish were subjected to a metabolic trial consisting of a 6-day residence in deuterated water (2H2O). Measurements of blood glucose and hepatic glycogen 2H-enrichments through Nuclear Magnetic Resonance, complemented by mRNA levels of key-enzymes for intermediary metabolism were used to evaluate the catabolic pathways of dietary glycerol. Dietary glycerol had no impact on plasma glucose, but hepatic glycogen levels increased significantly with increasing dietary glycerol levels in both species. While trout was able to regulate circulating glycerol plasma, seabass presented elevated levels on the glycerol-supplemented diets. Despite revealing some significant differences between sampling time (6 and 24 h), none of the enzymes’ mRNA levels responded to the dietary treatment. In trout, the main source of blood glucose was not labeled with 2H (~60%, likely from diet) while other contributors did not differ with glycerol supplementation. In seabass, the unlabeled contribution was approximately half of that observed in trout (~30%), accompanied by a significant increase of gluconeogenic contributions at the triose-phosphate level to the blood glucose with increasing dietary glycerol. In trout, labeling from 2H2O into hepatic glycogen revealed significant differences, with the contribution from the indirect pathway at the triose-phosphate level increasing with increased dietary glycerol. No such differences were found in seabass’ glycogen pool. These findings suggest that fish species are able to retain, catabolize glycerol and incorporate it into carbohydrates. The gluconeogenic utilization of exogenous glycerol differed between species and affected the synthesis of hepatic glycogen in trout and the appearance of blood glucose in seabass.
The W-Acidic Motif of Histidine Kinase WalK Is Required for Signaling and Transcriptional Regulation in Streptococcus mutans
Authors: Kong Lingyuan , Su Mingyang , Sang Jiayan , Huang Shanshan , Wang Min , Cai Yongfei , Xie Mingquan , Wu Jun , Wang Shida , Foster Simon J. , Zhang Jiaqin , Han Aidong
Published: April 2022

In Streptococcus mutans, we find that the histidine kinase WalK possesses the longest C-terminal tail (CTT) among all 14 TCSs, and this tail plays a key role in the interaction of WalK with its response regulator WalR. We demonstrate that the intrinsically disordered CTT is characterized by a conserved tryptophan residue surrounded by acidic amino acids. Mutation in the tryptophan not only disrupts the stable interaction, but also impairs the efficient phosphotransferase and phosphatase activities of WalRK. In addition, the tryptophan is important for WalK to compete with DNA containing a WalR binding motif for the WalR interaction. We further show that the tryptophan is important for in vivo transcriptional regulation and bacterial biofilm formation by S. mutans. Moreover, Staphylococcus aureus WalK also has a characteristic CTT, albeit relatively shorter, with a conserved W-acidic motif, that is required for the WalRK interaction in vitro. Together, these data reveal that the W-acidic motif of WalK is indispensable for its interaction with WalR, thereby playing a key role in the WalRK-dependent signal transduction, transcriptional regulation and biofilm formation.
Virulence and Host Range of Fungi Associated With the Invasive Plant Ageratina adenophora
Authors: Chen Lin , Yang Ai-Ling , Li Yu-Xuan , Zhang Han-Bo
Published: April 2022

To determine whether disease-mediated invasion of exotic plants can occur and whether this increases the risk of disease transmission in local ecosystems, it is necessary to characterize the species composition and host range of pathogens accumulated in invasive plants. In this study, we found that Didymellaceae, a family containing economically important plant fungal pathogens, is commonly associated with the invasive plantAgeratina adenophora. Accordingly, we characterized its phylogenetic position through multi-locus phylogenetic analysis, as well as its environmental distribution, virulence, and host range. The results indicated that 213 fungal collections were from 11 genera in Didymellaceae, ten of which are known, and one is potentially new.Didymella,Epicoccum,Remotididymella,andMesophomawere the dominant genera, accounting for 93% of total isolates. The virulence and host ranges of these fungi were related to their phylogenetic relationship.Boeremia exigua, Epicoccum latusicollum,andE. sorghinumwere found to be strongly virulent toward all tested native plants as well as towardA. adenophora;M. speciosaandM. ageratinaewere weakly virulent toward native plants but strongly virulent towardA. adenophora, thus displaying a narrow host range. Co-evolution analysis showed no strong phylogenetical signal between Didymellaceae and host plants. Isolates S188 and Y122 (belonging toM. speciosaandM. ageratinae, respectively) showed strong virulence towardA. adenophorarelative to native plants, highlighting their potential as biocontrol agents forA. adenophorainvasion. This study provides new insights into the understanding of the long-term ecological consequences of disease transmission driven by plant invasion.
Pharmaceutical Pollution in Aquatic Environments: A Concise Review of Environmental Impacts and Bioremediation Systems
Authors: Ortúzar Maite , Esterhuizen Maranda , Olicón-Hernández Darío Rafael , González-López Jesús , Aranda Elisabet
Published: April 2022

The presence of emerging contaminants in the environment, such as pharmaceuticals, is a growing global concern. The excessive use of medication globally, together with the recalcitrance of pharmaceuticals in traditional wastewater treatment systems, has caused these compounds to present a severe environmental problem. In recent years, the increase in their availability, access and use of drugs has caused concentrations in water bodies to rise substantially. Considered as emerging contaminants, pharmaceuticals represent a challenge in the field of environmental remediation; therefore, alternative add-on systems for traditional wastewater treatment plants are continuously being developed to mitigate their impact and reduce their effects on the environment and human health. In this review, we describe the current status and impact of pharmaceutical compounds as emerging contaminants, focusing on their presence in water bodies, and analyzing the development of bioremediation systems, especially mycoremediation, for the removal of these pharmaceutical compounds with a special focus on fungal technologies.
Identification of Prognostic Biomarkers in Patients With Malignant Rhabdoid Tumor of the Kidney Based on mTORC1 Signaling Pathway-Related Genes
Authors: Zhanghuang Chenghao , Yao Zhigang , Tang Haoyu , Zhang Kun , Wu Chengchuang , Li Li , Xie Yucheng , Yang Zhen , Yan Bing
Published: April 2022

Background: Malignant rhabdoid tumor of the kidney (MRTK) is an infrequent malignant tumor in childhood, accounting for approximately 2% of all childhood kidney tumors. Although the development of current treatments, the overall survival (OS) rate of MRTK patients is only 25%. The aim of this research was to explore the prognostic value of genes associated with the mTORC1 signaling pathway in MRTK.

Methods: The transcriptome data of MRTK samples were downloaded from the TARGET database. The 200 genes of HALLMARK_MTORC1_SIGNALING were downloaded from the Molecular Signatures Database (MSigDB). Furthermore, we applied gene set variation analysis (GSVA) to screen differentially expressed gene sets between the MRTK and normal samples. The 200 genes were combined with differentially expressed genes (DEGs) identified from differentially expressed gene sets. Then, a gene signature of mTORC1 pathway-related genes (mTRGs) was constructed in MRTK. The molecular mechanism of prognostic factors in MRTK was further analyzed using gene set enrichment analysis (GSEA). The target drugs based on these prognostic factors were explored from The Comparative Toxicogenomics Database (CTD). Moreover, six paired fresh tumor tissues and paraneoplastic tissues from children with MRTK were collected to validate the expressions of P4HA1, MLLT11, AURKA, and GOT1 in clinical samples via real-time fluorescence quantitative PCR and Western blot.

Results: A four-gene signature (P4HA1, MLLT11, AURKA, and GOT1) related to the mTORC1 pathway was developed in MRTK, which divided the MRTK patients into high-risk and low-risk groups. The patients with high-risk scores were strongly associated with reduced OS. Receiver operating characteristic (ROC) analysis indicated a good prediction performance of the four biomarker signatures. GSEA revealed that the mTOR signaling pathway was significantly enriched. The risk score was demonstrated to be an independent predictor for MRTK outcome. According to the correlation of tumor stem cell index and prognostic factors, the target drugs were obtained for the treatment of MRTK patients. Furthermore, the expressions of RT-qPCR and Western blot were consistent with RNA-sequencing data such that their expressions were significantly elevated in tumor tissues.

Conclusion: A total of four genes (P4HA1, MLLT11, AURKA, and GOT1) were screened as prognostic markers, further providing a new understanding for the treatment of patients with MRTK.
A Computational Model for the PLP-Dependent Enzyme Methionine γ-Lyase
Authors: Chen Xingyu , Briozzo Pierre , Machover David , Simonson Thomas
Published: April 2022

Pyridoxal-5′-phosphate (PLP) is a cofactor in the reactions of over 160 enzymes, several of which are implicated in diseases. Methionine γ-lyase (MGL) is of interest as a therapeutic protein for cancer treatment. It binds PLP covalently through a Schiff base linkage and digests methionine, whose depletion is damaging for cancer cells but not normal cells. To improve MGL activity, it is important to understand and engineer its PLP binding. We develop a simulation model for MGL, starting with force field parameters for PLP in four main states: two phosphate protonation states and two tautomeric states, keto or enol for the Schiff base moiety. We used the force field to simulate MGL complexes with each form, and showed that those with a fully-deprotonated PLP phosphate, especially keto, led to the best agreement with MGL structures in the PDB. We then confirmed this result through alchemical free energy simulations that compared the keto and enol forms, confirming a moderate keto preference, and the fully-deprotonated and singly-protonated phosphate forms. Extensive simulations were needed to adequately sample conformational space, and care was needed to extrapolate the protonation free energy to the thermodynamic limit of a macroscopic, dilute protein solution. The computed phosphate pKa was 5.7, confirming that the deprotonated, −2 form is predominant. The PLP force field and the simulation methods can be applied to all PLP enzymes and used, as here, to reveal fine details of structure and dynamics in the active site.
A Novel Blood Inflammatory Indicator for Predicting Deterioration Risk of Mild Traumatic Brain Injury
Authors: Ge Xintong , Zhu Luoyun , Li Meimei , Li Wenzhu , Chen Fanglian , Li Yongmei , Zhang Jianning , Lei Ping
Published: April 2022

Mild traumatic brain injury (mTBI) has a relatively higher incidence in aging people due to walking problems. Cranial computed tomography and magnetic resonance imaging provide the standard diagnostic tool to identify intracranial complications in patients with mTBI. However, it is still necessary to further explore blood biomarkers for evaluating the deterioration risk at the early stage of mTBI to improve medical decision-making in the emergency department. The activation of the inflammatory response is one of the main pathological mechanisms leading to unfavorable outcomes of mTBI. As complete blood count (CBC) analysis is the most extensively used laboratory test in practice, we extracted clinical data of 994 patients with mTBI from two large clinical cohorts (MIMIC-IV and eICU-CRD) and selected inflammation-related indicators from CBC analysis to investigate their relationship with the deterioration after mTBI. The combinatorial indices neutrophil-to-lymphocyte ratio (NLR), red cell distribution width-to-platelet ratio (RPR), and NLR times RPR (NLTRP) were supposed to be potential risk predictors, and the data from the above cohorts were integratively analyzed using our previously reported method named MeDICS. We found that NLR, RPR, and NLTRP levels were higher among deteriorated patients than non-deteriorated patients with mTBI. Besides, high NLTRP was associated with increased deterioration risk, with the odds ratio increasing from NLTRP of 1–2 (2.69, 1.48–4.89) to > 2 (4.44, 1.51–13.08), using NLTRP of 0–1 as the reference. NLTRP had a moderately good prognostic performance with an area under the ROC curve of 0.7554 and a higher prediction value than both NLR and RPR, indicated by the integrated discrimination improvement index. The decision curve analysis also showed greater clinical benefits of NLTRP than NLR and RPR in a large range of threshold probabilities. Subgroup analysis further suggested that NLTRP is an independent risk factor for the deterioration after mTBI. In addition, in vivo experiments confirmed the association between NLTRP and neural/systemic inflammatory response after mTBI, which emphasized the importance of controlling inflammation in clinical treatment. Consequently, NLTRP is a promising biomarker for the deterioration risk of mTBI. It can be used in resource-limited settings, thus being proposed as a routinely available tool at all levels of the medical system.
2-Methoxyestradiol Alleviates Neuroinflammation and Brain Edema in Early Brain Injury After Subarachnoid Hemorrhage in Rats
Authors: Hu Qiang , Du Quan , Yu Wenhua , Dong Xiaoqiao
Published: April 2022

Objective

Numerous studies have shown that neuroinflammation and brain edema play an important role in early brain injury (EBI) after subarachnoid hemorrhage (SAH). 2-Methoxyestradiol (2-ME) has been shown to have anti-inflammatory and anti-angiogenic effects. This study aimed to investigate the effects of 2-ME on neuroinflammation and brain edema after SAH and its underlying mechanism of action.

Methods

Rats were used to produce an endovascular puncture model of SAH. 2-ME or the control agent was injected intraperitoneally 1 h after SAH induction. At 24 h after surgery, the neurological score, SAH grading, brain water content, and blood–brain barrier (BBB) permeability were examined. The microglial activation level in the rat brain tissue was determined using immunofluorescence staining, whereas the cell apoptosis in the rat brain tissue was assessed using terminal deoxynucleotidyl transferase dUTP nick-end labeling assay, the levels of Interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α were measured by enzyme linked immunosorbent assay, and the expression levels of ZO-1, occludin, hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), and matrix metallopeptidase (MMP)-9 in the rat brain tissue were determined using western blotting.

Results

Twenty-four hours after SAH, brain water content, BBB permeability, microglial activation, and cell apoptosis were significantly increased, whereas neurological function deteriorated significantly in rats. Treatment with 2-ME significantly decreased brain water content, BBB permeability, microglial cell activation, and cell apoptosis and improved neurological dysfunction in rats. Treatment with 2-ME reduced the expression levels of inflammatory factors (IL-1β, IL-6, and TNF-α), which were significantly elevated 24 h after SAH. Treatment with 2-ME alleviated the disruption of tight junction proteins (ZO-1 and occludin), which significantly decreased 24 h after SAH. To further determine the mechanism of this protective effect, we found that 2-ME inhibited the expression of HIF-1α, MMP-9, and VEGF, which was associated with the inflammatory response to EBI and BBB disruption after SAH.

Conclusion

2-ME alleviated neuroinflammation and brain edema as well as improved neurological deficits after SAH in rats. The neuroprotective effect of 2-ME on EBI after SAH in rats may be related to the inhibition of neuroinflammation and brain edema.
Imaging Insights of Isolated Idiopathic Dystonia: Voxel-Based Morphometry and Activation Likelihood Estimation Studies
Authors: Wu Yunhao , Zhang Chao , Li Yufei , Feng Jie , Zhang Ming , Li Hongxia , Wang Tao , Zhang Yingying , Jin Zhijia , Zhang Chencheng , Zhang Yuyao , Li Dianyou , Wu Yiwen , Wei Hongjiang , Sun Bomin
Published: April 2022

The understanding of brain structural abnormalities across different clinical forms of dystonia and their contribution to clinical characteristics remains unclear. The objective of this study is to investigate shared and specific gray matter volume (GMV) abnormalities in various forms of isolated idiopathic dystonia. We collected imaging data from 73 isolated idiopathic dystonia patients and matched them with healthy controls to explore the GMV alterations in patients and their correlations with clinical characteristics using the voxel-based morphometry (VBM) technique. In addition, we conducted an activation likelihood estimation (ALE) meta-analysis of previous VBM studies. Our study demonstrated widespread morphometry alterations in patients with idiopathic dystonia. Multiple systems were affected, which mainly included basal ganglia, sensorimotor, executive control, and visual networks. As the result of the ALE meta-analysis, a convergent cluster with increased GMV was found in the left globus pallidus. In subgroup VBM analyses, decreased putamen GMV was observed in all clinic forms, while the increased GMV was observed in parahippocampal, lingual, and temporal gyrus. GD demonstrated the most extensive GMV abnormalities in cortical regions, and the aberrant GMV of the posterior cerebellar lobe was prominent in CD. Moreover, trends of increased GMV regions of the left precuneus and right superior frontal gyrus were demonstrated in the moderate-outcome group compared with the superior-outcome group. Results of our study indicated shared pathophysiology of the disease-centered on the dysfunction of the basal ganglia-thalamo-cortical circuit, impairing sensorimotor integration, high-level motor execution, and cognition of patients. Dysfunction of the cerebello-thalamo-cortical circuit could also be involved in CD especially. Finally, the frontal-parietal pathway may act as a potential marker for predicting treatment outcomes such as deep brain stimulation.
Clinical Study of 30 Novel KCNQ2 Variants/Deletions in KCNQ2-Related Disorders
Authors: Xiao Tiantian , Chen Xiang , Xu Yan , Chen Huiyao , Dong Xinran , Yang Lin , Wu Bingbing , Chen Liping , Li Long , Zhuang Deyi , Chen Dongmei , Zhou Yuanfeng , Wang Huijun , Zhou Wenhao
Published: April 2022

Background

KCNQ2-related disorder is typically characterized as neonatal onset seizure and epileptic encephalopathy. The relationship between its phenotype and genotype is still elusive. This study aims to provide clinical features, management, and prognosis of patients with novel candidate variants of the KCNQ2 gene.

Methods

We enrolled patients with novel variants in the KCNQ2 gene from the China Neonatal Genomes Project between January 2018 and January 2021. All patients underwent next-generation sequencing tests and genetic data were analyzed by an in-house pipeline. The pathogenicity of variants was classified according to the guideline of the American College of Medical Genetics. Each case was evaluated by two geneticists back to back. Patients' information was acquired from clinical records.

Results

A total of 30 unrelated patients with novel variants in the KCNQ2 gene were identified, including 19 patients with single-nucleotide variants (SNVs) and 11 patients with copy number variants (CNVs). For the 19 SNVs, 12 missense variants and 7 truncating variants were identified. Of them, 36.8% (7/19) of the KCNQ2 variants were located in C-terminal regions, 15.7% (3/19) in segment S2, and 15.7% (3/19) in segment S4. Among them, 18 of 19 patients experienced seizures in the early neonatal period. However, one patient presented neurodevelopmental delay (NDD) as initial phenotype when he was 2 months old, and he had severe NDD when he was 3 years old. This patient did not present seizure but had abnormal electrographic background activity and brain imaging. Moreover, for the 11 patients with CNVs, 20q13.3 deletions involving EEF1A2, KCNQ2, and CHRNA4 genes were detected. All of them presented neonatal-onset seizures, responded to antiepileptic drugs, and had normal neurological development.

Conclusion

In this study, patients with novel KCNQ2 variants have variable phenotypes, whereas patients with 20q13.3 deletion involving EEF1A2, KCNQ2, and CHRNA4 genes tend to have normal neurological development.
The Burden of Undernutrition and Its Associated Factors Among Children Below 5 Years of Age in Bambao Region, Comoros
Authors: Ahamada Hadji , Sunguya Bruno F.
Published: April 2022

Background

Undernutrition remains a major public health problem in low- and middle-income countries and Comoros is no exception. This study aimed to examine the prevalence and identify the risk factors of undernutrition among children under-five years in Bambao region, Comoros.

Methods

This cross-sectional study was conducted in Bambao region among 837 under-five years and their caregivers. Analyses were conducted using both descriptive and logistic regression to examine the magnitude and factors associated with stunting, wasting and underweight.

Results

Prevalence of stunting, wasting and underweight were 21.6, 13.7, and 13.6% respectively. Factors associated with stunting were caregiver's secondary education level compared to no education (AOR = 1.89, 95% CI: 1.04–3.43, P < 0.04), age of child between 13–24 months compared to 0–12 months (AOR = 2.69, 95% CI: 1.44–5.01, P < 0.001), and food insecurity (AOR = 2.55, 95% CI: 1.20–5.41, P < 0.02). Children aged 25–59 months were 78% less likely to have wasting compared to those with 0–12 months (AOR = 0.22, 95% CI: 0.10–0.51, P < 0.001). Wasting was also associated with food insecurity (AOR = 2.70, 95% CI: 1.12–6.49, P < 0.03), and low birthweight (AOR = 3.21, 95% CI: 1.73–5.94, P < 0.001). Children aged between 25–59 months were 86% less likely to have underweight compared to those aged 0–12 months (AOR = 0.14, 95% CI: 0.06–0.36, P < 0.001). Food insecurity (AOR = 2.65, 95% CI: 1.08–6.54, P < 0.03), low birthweight (AOR = 3.15, 95% CI: 1.67–5.93, P < 0.001), and non-exclusively breastfeeding (AOR = 2.37, 95% CI: 1.15–4.90, P < 0.02) were also associated with underweight.

Conclusion

More than one in five children under-five is stunted in Bambao region, Comoros. Moreover, more than 13% are underweight or wasted calling for streamlined efforts to address poor feeding practices, food insecurity, low birthweight, and socio-demographic disadvantages in this and other areas with similar context.
Optimisation and Application of a Novel Method to Identify Bacteriophages in Maternal Milk and Infant Stool Identifies Host-Phage Communities Within Preterm Infant Gut
Authors: Young Gregory R. , Yew Wen C. , Nelson Andrew , Bridge Simon H. , Berrington Janet E. , Embleton Nicholas D. , Smith Darren L.
Published: April 2022

Human milk oligosaccharides, proteins, such as lactoferrin, and bacteria represent just some of the bioactive components of mother's breast milk (BM). Bacteriophages (viruses that infect bacteria) are an often-overlooked component of BM that can cause major changes in microbial composition and metabolism. BM bacteriophage composition has been explored in term and healthy infants, suggesting vertical transmission of bacteriophages occurs between mothers and their infants. Several important differences between term and very preterm infants (<30 weeks gestational age) may limit this phenomenon in the latter. To better understand the link between BM bacteriophages and gut microbiomes of very preterm infants in health and disease, standardised protocols are required for isolation and characterisation from BM. In this study, we use isolated nucleic acid content, bacteriophage richness and Shannon diversity to validate several parameters applicable during bacteriophage isolation from precious BM samples. Parameters validated include sample volume required; centrifugal sedimentation of microbes; hydrolysis of milk samples with digestive enzymes; induction of temperate bacteriophages and concentration/purification of isolated bacteriophage particles in donor milk (DM). Our optimised method enables characterisation of bacteriophages from as little as 0.1 mL BM. We identify viral families that were exclusively identified with the inclusion of induction of temperate bacteriophages (Inoviridae) and hydrolysis of milk lipid processes (IridoviridaeandBaculoviridae). Once applied to a small clinical cohort we demonstrate the vertical transmission of bacteriophages from mothers BM to the gut of very preterm infants at the species level. This optimised method will enable future research characterising the bacteriophage composition of BM in very preterm infants to determine their clinical relevance in the development of a healthy preterm infant gut microbiome.
Visual Function Score: A New Clinical Tool to Assess Visual Function and Detect Visual Disorders in Children
Authors: Signorini Sabrina , Luparia Antonella , Cappagli Giulia , Perotto Eleonora , Antonini Mauro , Morelli Federica , Aprile Giorgia , Ballante Elena , Figini Silvia , Borgatti Renato ,   VFS Study Group
Published: April 2022

Introduction

A comprehensive assessment of visual functioning at an early age is important not only for identifying and defining visual impairment but also for planning personalized rehabilitation programs based on the visual diagnosis. Since existing tools to evaluate visual functioning present some important limitations (e.g., they are based on qualitative reports, they do not take into account environmental adaptations of visual testing or they have not been formally validated as clinical instruments), the present work has the main aim to propose a new clinical tool (Visual Function Score, VFS) to detect and define visual disorders at an early age.

Methods

The Visual Function Score was administered to one hundred visually impaired children (age range 4 months to 17.75 years old) in the form of a professional-reported protocol for a total of 51 items, each of which is assigned a score from 1 to 9 (or from 0 to 9 in some specific cases). The VFS produces three sub-scores and a global score (from 0 to 100), resulting in a quantitative evaluation of visual functioning.

Results

The VFS can detect the well-known differences between different types of visual impairment (cerebral, oculomotor, and peripheral or grouped as central and peripheral) and takes into account different environments in the definition of a quantitative score of visual functioning.

Discussion

Overall, the use of a quantitative tool to evaluate visual functions and functional vision such as the VFS would be fundamental to monitor the progresses of patients over time in response to rehabilitation interventions.
Modified Adenosines Sensitize Glioblastoma Cells to Temozolomide by Affecting DNA Methyltransferases
Authors: Proto Maria Chiara , Fiore Donatella , Piscopo Chiara , Laezza Chiara , Bifulco Maurizio , Gazzerro Patrizia
Published: April 2022

Glioblastoma (GBM) is the most common and lethal primary malignant brain tumor, and due to its unique features, its management is certainly one of the most challenging ones among all cancers. N6-isopentenyladenosine (IPA) and its analog N6-benzyladenosine (N6-BA) are modified nucleosides endowed with potent antitumor activity on different types of human cancers, including GBM. Corroborating our previous finding, we demonstrated that IPA and N6-BA affect GBM cell line proliferation by modulating the expression of the F-box WD repeat domain-containing-7 (FBXW7), a tumor suppressor with a crucial role in the turnover of many proteins, such as SREBPs and Mcl1, involved in malignant progression and chemoresistance. Luciferase assay revealed that IPA-mediated upregulation of FBXW7 translates in transcriptional inactivation of its oncogenic substrates (Myc, NFkB, or HIF-1α). Moreover, downregulating MGMT expression, IPA strongly enhances the killing effect of temozolomide (TMZ), producing a favorable sensitizing effect starting from a concentration range much lower than TMZ EC50. Through DNA methyltransferase (DNMT) activity assay, analysis of the global DNA methylation, and the histone modification profiles, we demonstrated that the modified adenosines behave similar to 5-AZA-dC, known DNMT inhibitor. Overall, our results provide new perspectives for the first time, suggesting the modified adenosines as epigenetic tools able to improve chemo- and radiotherapy efficacy in glioblastoma and potentially other cancers.
Cost-Effectiveness Analysis of Hepatic Arterial Infusion Chemotherapy of Infusional Fluorouracil, Leucovorin, and Oxaliplatin Versus Transarterial Chemoembolization in Patients With Large Unresectable Hepatocellular Carcinoma
Authors: Zhang Haixia , Zeng Xiaohui , Peng Ye , Tan Chongqing , Wan Xiaomin
Published: April 2022

Purpose: The purpose of this study was to evaluate a cost-effectiveness analysis of hepatic arterial infusion chemotherapy with infusional fluorouracil, leucovorin, and oxaliplatin (FOLFOX-HAIC) as the first-line treatment for patients with large unresectable hepatocellular carcinoma (HCC) compared with transarterial chemoembolization (TACE).

Methods: A Markov model was constructed to simulate the first-line treatment, disease recurrence, and survival of patients with large unresectable HCC. Transition probabilities were based on clinical trial data. The costs and health utilities were derived from the public literature. The outputs were total cost, quality-adjusted life year (QALY), and incremental cost-effectiveness ratios (ICER). One-way and probabilistic sensitivity analyses were performed to examine model uncertainty. We also performed subgroup analyses.

Results: The results of the base case analysis found that FOLFOX-HAIC increased overall costs by $9,381 and improved effectiveness by 1.01 QALYs compared with TACE. The one-way sensitivity analysis showed that the hazard ratio of progression-free survival and overall survival for FOLFOX-HAIC relative to TACE had the greatest impact on the ICER. Probabilistic sensitivity analysis found that the probability of FOLFOX-HAIC treatment being cost-effective was 99.54% at the willingness-to-pay threshold of $30,552/QALY. Patients in most subgroups favored FOLFOX-HAIC treatment because it had a more than 50% probability of being cost-effective than TACE, except for patients with negative hepatitis B infection.

Conclusion: In conclusion, our study found that the FOLFOX-HAIC was a cost-effective option compared to TACE for patients with large unresectable HCC in China.
Echinacea Purpurea For the Long-Term Prevention of Viral Respiratory Tract Infections During Covid-19 Pandemic: A Randomized, Open, Controlled, Exploratory Clinical Study
Authors: Kolev Emil , Mircheva Lilyana , Edwards Michael R. , Johnston Sebastian L. , Kalinov Krassimir , Stange Rainer , Gancitano Giuseppe , Berghe Wim Vanden , Kreft Samo
Published: April 2022

SARS-CoV-2 vaccination is effective in preventing severe Covid-19, but efficacy in reducing viral load and transmission wanes over time. In addition, the emergence of novel SARS-CoV-2 variants increases the threat of uncontrolled dissemination and additional antiviral therapies are urgently needed for effective containment. In previous in vitro studies Echinacea purpurea demonstrated strong antiviral activity against enveloped viruses, including SARS-CoV-2. In this study, we examined the potential of Echinacea purpurea in preventing and treating respiratory tract infections (RTIs) and in particular, SARS-CoV-2 infections. 120 healthy volunteers (m,f, 18—75 years) were randomly assigned to Echinacea prevention or control group without any intervention. After a run-in week, participants went through 3 prevention cycles of 2, 2 and 1 month with daily 2,400 mg Echinacea purpurea extract (Echinaforce®, EF). The prevention cycles were interrupted by breaks of 1 week. Acute respiratory symptoms were treated with 4,000 mg EF for up to 10 days, and their severity assessed via a diary. Naso/oropharyngeal swabs and venous blood samples were routinely collected every month and during acute illnesses for detection and identification of respiratory viruses, including SARS-CoV-2 via RT-qPCR and serology. Summarized over all phases of prevention, 21 and 29 samples tested positive for any virus in the EF and control group, of which 5 and 14 samples tested SARS-CoV-2 positive (RR = 0.37, Chi-square test, p = 0.03). Overall, 10 and 14 symptomatic episodes occurred, of which 5 and 8 were Covid-19 (RR = 0.70, Chi-square test, p > 0.05). EF treatment when applied during acute episodes significantly reduced the overall virus load by at least 2.12 log10 or approx. 99% (t-test, p < 0.05), the time to virus clearance by 8.0 days for all viruses (Wilcoxon test, p = 0.02) and by 4.8 days for SARS-CoV-2 (p > 0.05) in comparison to control. Finally, EF treatment significantly reduced fever days (1 day vs 11 days, Chi-square test, p = 0.003) but not the overall symptom severity. There were fewer Covid-19 related hospitalizations in the EF treatment group (N = 0 vs N = 2). EF exhibited antiviral effects and reduced the risk of viral RTIs, including SARS-CoV-2. By substantially reducing virus loads in infected subjects, EF offers a supportive addition to existing mandated treatments like vaccinations. Future confirmatory studies are warranted.
Berberine Attenuates Cell Motility via Inhibiting Inflammation-Mediated Lysyl Hydroxylase-2 and Glycolysis
Authors: Du Yishan , Khan Muhammad , Fang Nana , Ma Fang , Du Hongzhi , Tan Zhenya , Wang Hua , Yin Shi , Wei Xiaohui
Published: April 2022

Lysyl hydroxylase-2 (LH2) involves in the hydroxylation of telopeptide lysine residues during collagen deposition. Recent studies indicate that interleukin (IL)-6 generated by the chronic inflammation disease may trigger the LH2 expression to accelerate cell motility. Berberine is the alkaloid derived from the traditional Chinese medicine Coptis chinensis, which displays potential anti-inflammatory activity in multiple diseases. The anti-inflammatory activity of berberine has been confirmed by reducing proinflammatory cytokines such as IL-6, IL-8, and IFN-γ. However, whether and how berberine inhibits cellular motility against metastatic spread in triple-negative breast cancer (TNBC) has not been demonstrated, and the underlying mechanism remains unclear. We investigated the effects of berberine on the inflammatory cytokine secretion, cell proliferation, and migration in vitro and further explored the effect of berberine on growth and metastasis in vivo. Berberine restrained TNBC cell proliferation, motility, and glycolysis process in a dose-dependent way. The secretion of IL-6 was abrogated by berberine in TNBC cells, and IL-6-stimulated cell migration was inhibited by berberine. Mechanistically, berberine remarkably suppressed LH2 expression at both mRNA and protein levels. LH2 depletion led to decreasing the antimotility effect of berberine, and this phenomenon was related to the suppressed glycolysis after LH2 inhibition. Conversely, ectopic restoration of LH2 could further increase the antimotility effect of berberine. Moreover, berberine was confirmed to inhibit cell growth and motility in vivo, and the expression of LH2 and glycolytic enzymes was also blocked by berberine in vivo. Collectively, this study indicated that berberine could be a promising therapeutic drug via regulating LH2 for TNBC.
Dysregulation of Vesicular Glutamate Transporter VGluT2 via BDNF/TrkB Pathway Contributes to Morphine Tolerance in Mice
Authors: He Liqiong , Xu Wei , Zhang Chengliang , Ding Zhuofeng , Guo Qulian , Zou Wangyuan , Wang Jian
Published: April 2022

Morphine is widely used in the treatment of moderate to severe pain. Long-term use of morphine leads to various adverse effects, such as tolerance and hyperalgesia. Vesicular glutamate transporter 2 (VGluT2) accumulates glutamate into synaptic vesicles and plays multiple roles in the central nervous system. However, the specific role of VGluT2 in morphine tolerance has not been fully elucidated. Here, we investigated the regulatory role of VGluT2 in morphine tolerance and assessed the potential role of the brain-derived neurotrophic factor (BDNF)/tyrosine kinase B (TrkB) pathway in VGluT2 mediated morphine antinociceptive tolerance in mice. In the present study, we found that VGluT2 is upregulated in the spinal cord after the development of morphine tolerance. Furthermore, inhibition of VGluT2 with its antagonist (Chicago sky blue 6 B, CSB6B) or knockdown of VGluT2 by lentivirus restored the analgesic effect of morphine, suppressed the activation of astrocytes and microglia, and decreased glial-derived pro-inflammatory cytokines. Overexpression of VGluT2 by lentivirus facilitated morphine tolerance and mechanical hyperalgesia. In addition, we found the expression of BDNF is correlated with VGluT2 expression in the spinal cord after chronic morphine administration. Intrathecal injection of the BDNF/TrkB pathway antagonist K252a attenuated the development of morphine tolerance and decreased the expression of VGluT2 in the spinal cord, which suggested the BDNF/TrkB pathway participates in the regulation of VGluT2 in morphine tolerance. This study elucidates the functional capability of VGluT2 in modulating morphine tolerance and identifies a novel mechanism and promising therapeutic target for morphine tolerance.
Dimethyl Fumarate Ameliorates Doxorubicin-Induced Cardiotoxicity By Activating the Nrf2 Pathway
Authors: Hu Xiaoliang , Li Cheng , Wang Qian , Wei Zhixing , Chen Taizhong , Wang Yuepeng , Li Yigang
Published: April 2022

Doxorubicin (DOX) is limited in clinical application because of its cardiotoxicity. Oxidative stress and apoptosis are crucial in DOX-induced cardiac injury. Dimethyl fumarate (DMF) is an FDA-approved oral drug with powerful effects to reduce oxidative stress and apoptosis through the Nrf2 pathway. This study was aimed to determine whether DMF can protect against DOX-induced cardiac injury. We used both neonatal rat cardiomyocytes (NRCMs) in vitro and DOX-induced cardiac toxicity in vivo to explore the effects of DMF. The results showed that DMF significantly improved cell viability and morphology in NRCMs. In addition, DMF alleviated DOX-induced cardiac injury in rats, as evidenced by decreased CK-MB, LDH levels, improved survival rates, cardiac function, and pathological changes. Moreover, DMF significantly inhibited cardiac oxidative stress by reducing MDA levels and increasing GSH, SOD, and GSH-px levels. And DMF also inhibited DOX-induced cardiac apoptosis by modulating Bax, Bcl-2 and cleaved caspase-3 expression. Moreover, DMF exerted its protective effects against DOX by promoting Nrf2 nuclear translocation, which activated its downstream antioxidant gene Hmox1. Silencing of Nrf2 attenuated the protective effects of DMF in NRCMs as manifested by increased intracellular oxidative stress, elevated apoptosis levels, and decreased cell viability. In addition, DMF showed no protective effects on the viability of DOX-treated tumor cells, which suggested that DMF does not interfere with the antitumor effect of DOX in vitro. In conclusion, our data confirmed that DMF alleviated DOX-induced cardiotoxicity by regulating oxidative stress and apoptosis through the Nrf2 pathway. DMF may serve as a new candidate to alleviate DOX-related cardiotoxicity in the future.
Semi-Synthetic Analogues of Cryptolepine as a Potential Source of Sustainable Drugs for the Treatment of Malaria, Human African Trypanosomiasis, and Cancer
Authors: Abacha Yabalu Z. , Forkuo Arnold Donkor , Gbedema Stephen Y. , Mittal Nimisha , Ottilie Sabine , Rocamora Frances , Winzeler Elizabeth A. , van Schalkwyk Donelly A. , Kelly John M. , Taylor Martin C. , Reader Janette , Birkholtz Lyn-Marie , Lisgarten David R. , Cockcroft Jeremy K. , Lisgarten John N. , Palmer Rex A. , Talbert Rosemary C. , Shnyder Steven D. , Wright Colin W.
Published: April 2022

The prospect of eradicating malaria continues to be challenging in the face of increasing parasite resistance to antimalarial drugs so that novel antimalarials active against asexual, sexual, and liver-stage malaria parasites are urgently needed. In addition, new antimalarials need to be affordable and available to those most in need and, bearing in mind climate change, should ideally be sustainable. The West African climbing shrub Cryptolepis sanguinolenta is used traditionally for the treatment of malaria; its principal alkaloid, cryptolepine (1), has been shown to have antimalarial properties, and the synthetic analogue 2,7-dibromocryptolepine (2) is of interest as a lead toward new antimalarial agents. Cryptolepine (1) was isolated using a two-step Soxhlet extraction of C. sanguinolenta roots, followed by crystallization (yield 0.8% calculated as a base with respect to the dried roots). Semi-synthetic 7-bromo- (3), 7, 9-dibromo- (4), 7-iodo- (5), and 7, 9-dibromocryptolepine (6) were obtained in excellent yields by reaction of 1 with N-bromo- or N-iodosuccinimide in trifluoroacetic acid as a solvent. All compounds were active against Plasmodia in vitro, but 6 showed the most selective profile with respect to Hep G2 cells: P. falciparum (chloroquine-resistant strain K1), IC50 = 0.25 µM, SI = 113; late stage, gametocytes, IC50 = 2.2 µM, SI = 13; liver stage, P. berghei sporozoites IC50 = 6.13 µM, SI = 4.6. Compounds 3–6 were also active against the emerging zoonotic species P. knowlesi with 5 being the most potent (IC50 = 0.11 µM). In addition, 3–6 potently inhibited T. brucei in vitro at nM concentrations and good selectivity with 6 again being the most selective (IC50 = 59 nM, SI = 478). These compounds were also cytotoxic to wild-type ovarian cancer cells as well as adriamycin-resistant and, except for 5, cisplatin-resistant ovarian cancer cells. In an acute oral toxicity test in mice, 3–6 did not exhibit toxic effects at doses of up to 100 mg/kg/dose × 3 consecutive days. This study demonstrates that C. sanguinolenta may be utilized as a sustainable source of novel compounds that may lead to the development of novel agents for the treatment of malaria, African trypanosomiasis, and cancer.
Adolescent Binge Alcohol Enhances Early Alzheimer’s Disease Pathology in Adulthood Through Proinflammatory Neuroimmune Activation
Authors: Barnett Alexandra , David Emeraghi , Rohlman Aaron , Nikolova Viktoriya D. , Moy Sheryl S. , Vetreno Ryan P. , Coleman Leon G.
Published: April 2022

Epidemiological studies suggest that heavy alcohol use early in life is associated with increased risk for Alzheimer’s disease (AD). However, mechanisms connecting AD with alcohol use have not been identified. Both heavy alcohol use and AD feature increased proinflammatory signaling. Therefore, we hypothesized that adolescent binge ethanol would increase AD molecular and behavioral pathology in adulthood through proinflammatory signaling. The 3xTg-AD mouse model (APPSwe, tauP301, Psen1tm1Mpm) which features amyloid (Aβ) and tau pathology beginning at 6–12 months underwent adolescent intermittent ethanol (AIE, 5 g/kg/d, i.g., P25-55) with assessment of AD pathologic mediators at P200. A second group of mice received AIE +/− minocycline (30 mg/kg/d, IP) followed by behavioral testing in adulthood. Behavioral testing and age of testing included: locomotor activity and exploration (27–28 weeks), novel object recognition (NORT, 28-30 weeks), 3-chamber sociability and social memory (29–31 weeks), prepulse inhibition (PPI, 30–32 weeks), Morris Water Maze with reversal (MWM, 31–35 weeks), and Piezo sleep monitoring (35–37 weeks). We found that AIE increased levels of neurotoxic Aβ1–42 in adult female hippocampus as well as intraneuronal Aβ1–42 in amygdala and entorhinal cortex. Phosphorylated tau at residue Thr181 (p-tau-181) was also increased in female hippocampus by AIE. Several proinflammatory genes were persistently increased by AIE in the female hippocampus, including IL-1β, MCP-1, IL-6, and IFNα. Expression of these genes was strongly correlated with the levels of Aβ1–42 and p-tau-181 in hippocampus. AIE caused persistent decreases in locomotor activity (open-field and NORT habituation) and increased anxiety-like behavior (thigmotaxis) while reducing memory retention. Treatment with the anti-inflammatory compound minocycline during AIE blocked persistent increases in Aβ1–42 in amygdala and p-tau-181 in hippocampus, and prevented AIE-induced thigmotaxis and memory loss. Together, these data find that adolescent binge ethanol enhances AD molecular and behavioral pathology in adulthood through proinflammatory signaling. Blockade of proinflammatory signaling during ethanol exposure prevents ethanol-induced effects on pathologic accumulation of AD-associated proteins and persistent behavior changes relevant to human AD.