bluuebunny/crossref_metadata_2025 huggingface dataset:
Vitreoscilla Hemoglobin Improves Sophorolipid Production in Starmerella Bombicola O-13–1 Under Oxygen Limited Conditions
Authors: Li Jun-feng , Li Hong-fang , Yao Shu-min , Zhao Meng-juan , Dong Wen-xun , Liang Sheng-kang , Xu Xing-yong
Published: October 2021

Sophorolipids (SLs) are homologous microbial secondary metabolites produced by Starmerella bombicola and have been widely applied in many industrial fields. The biosynthesis of SLs is a highly aerobic process and is often limited by low dissolved oxygen (DO) levels. In this study, the Vitreoscilla hemoglobin (VHb) gene was transformed into S. bombicola O-13–1 by homologous recombination to alleviate oxygen limitation. VHb expression improved the intracellular oxygen utilization efficiency under either oxygen-rich or oxygen-limited conditions. In shake flask culture, the production of SLs was higher in the recombinant (VHb+) strain than in the wild-type (VHb−) strain, while the oxygen uptake rate of the recombinant (VHb+) strain was significantly lower than that of the wild-type (VHb−) strain. In a 5 L bioreactor, the production of SLs did not increase significantly, but the DO level in the fermentation broth of the VHb+ strain was 21.8% higher than that of VHb− strain under oxygen-rich conditions. Compared to wide-type strains (VHb−), VHb expression enhanced SLs production by 25.1% in the recombinants (VHb+) under oxygen-limited conditions. In addition, VHb expression raised the transcription levels of key genes involved in the electron transfer chain (NDH, SDH, COX), TCA cycle (CS, ICD, KDG1) and SL synthesis (CYP52M1 and UGTA1) in the recombinant (VHb+) strains. VHb expression in S. bombicola could enhance SLs biosynthesis and intracellular oxygen utilization efficiency by increasing ATP production and cellular respiration. Our findings highlight the potential use of VHb to improve the oxygen utilization efficiency of S. bombicola in the industrial-scale production of SLs using industrial and agricultural by-products like molasses and waste oil as fermentation feedstock.
Membrane Engineering for Biogas Valorization
Authors: Brunetti Adele , Barbieri Giuseppe
Published: October 2021

Membrane operations nowadays drive the innovative design of important separation, conversion, and upgrading processes, and contribute to realizing the main principles of “green process engineering” in various sectors. In this perspective, we propose the re-design of traditional plants for biogas upgrading and integrating and/or replacing conventional operations with innovative membrane units. Bio-digester gas streams contain valuable products such as biomethane, volatile organic compounds, and volatile fatty acids, whose recovery has important advantages for environment protection, energy saving, and waste valorization. Advanced membrane units can valorize biogas by separating its various components, and establishing environmentally friendly and small-scale energivorous novel separation processes enables researchers to pursue the requirements of circular economy.
Transition of Serotype 35B Pneumococci From Commensal to Prevalent Virulent Strain in Children
Authors: Fuji Naoko , Pichichero Michael , Ehrlich Rachel L. , Mell Joshua Chang , Ehrlich Garth D. , Kaur Ravinder
Published: October 2021

In our community-based prospective cohort study in young children, we observed a significant increase in pneumococcal serotype 35B nasopharyngeal (NP) commensal colonization during the 2011–2014 timeframe, but these strains were not associated with disease. Beginning in 2015 and continuing through to the present, the serotype 35B virulence changed, and it became the dominant bacteria isolated and associated with pneumococcal acute otitis-media (AOM) in our cohort. We performed comparative analyses of 250 35B isolates obtained from 140 children collected between 2006 and 2019. Changes in prevalence, clonal-complex composition, and antibiotic resistance were analyzed. Seventy-two (29%) of 35B isolates underwent whole-genome sequencing to investigate genomic changes associated with the shift in virulence that resulted in increased rates of 35B-associated AOM disease. 35B strains that were commensals and AOM disease-causing were mainly associated with sequence type (ST) 558. Antibiotic concentrations of β-lactams and ofloxacin necessary to inhibit growth of 35B strains rose significantly (2006–2019) (p<0.005). However, only isolates from the 35B/ST558 showed significant increases in MIC50 of penicillin and ofloxacin between the years 2006–2014 and 2015–2019 (p=0.007 and p<0.0001). One hundred thirty-eight SNPs located in 34 different genes were significantly associated with post-2015 strains. SNPs were found in nrdG (metal binding, 10%); metP and metN (ABC transporter, 9%); corA (Mg2+ transporter, 6%); priA (DNA replication, 5%); and on the enzymic gene ldcB (LD-carboxypeptidase, 3%). Pneumococcal serotype 35B strains was a common NP commensal during 2010–2014. In 2015, a shift in increasing number of AOM cases occurred in young children caused by 35B, that was associated with changes in genetic composition and antibiotic susceptibility.
Antimicrobial Activity of Phytic Acid: An Emerging Agent in Endodontics
Authors: Nassar Rania , Nassar Mohannad , Vianna Morgana E. , Naidoo Nerissa , Alqutami Fatma , Kaklamanos Eleftherios G. , Senok Abiola , Williams David
Published: October 2021

Background

Phytic acid (IP6) is a promising and emerging agent, and because of its unique structure and distinctive properties, it lends itself to several applications in dentistry. Recently, IP6 was proposed as a potential chelating agent in endodontics. However, there is limited knowledge regarding its antimicrobial and antibiofilm effectiveness. The aims of this study, were therefore to evaluate the antimicrobial and antibiofilm activities of IP6 against a range of microbial species and compare these with ethylenediaminetetraacetic acid (EDTA) and sodium hypochlorite (NaOCl). The contact time required for IP6 to exert its bactericidal effect on Enterococcus faecalis was also determined.

Methods

The inhibitory and biocidal activities of IP6, EDTA and NaOCl were assessed using a broth microdilution assay against 11 clinical and reference strains of bacteria and a reference strain of Candida albicans. The contact time required for various IP6 concentrations to eliminate planktonic cultures of E. faecalis was determined using a membrane filtration method according to BS-EN-1040:2005. IP6 bactericidal activity was also evaluated using fluorescent microscopy, and the antibiofilm activity of the test agents was also determined.

Results

IP6 was biocidal against all tested microorganisms. At concentrations of 0.5%, 1% and 2%, IP6 required 5 min to exert a bactericidal effect on E. faecalis, while 5% IP6 was bactericidal after 30 s. IP6 also eradicated biofilms of the tested microorganisms. In conclusion, IP6 had notable antimicrobial effects on planktonic and biofilm cultures and exhibited rapid bactericidal effects on E. faecalis. This research highlighted, for the first time the antimicrobial and antibiofilm properties of IP6, which could be exploited, not only in dental applications, but also other fields where novel strategies to counter antimicrobial resistance are required.
Trypanosoma cruzi trans-Sialidase as a Potential Vaccine Target Against Chagas Disease
Authors: da Costa Kelli Monteiro , Marques da Fonseca Leonardo , dos Reis Jhenifer Santos , Santos Marcos André Rodrigues da Costa , Previato José Osvaldo , Mendonça-Previato Lucia , Freire-de-Lima Leonardo
Published: October 2021

Chagas’ disease is caused by the protozoanTrypanosoma cruzi, described in the early 20thcentury by the Brazilian physician Dr. Carlos Chagas. There was a great amount of research devoted to diagnosis, treatment and prevention of the disease. One of the most important discoveries made since then, impacting the understanding of how the parasite interacts with the host’s immune system, was the description oftrans-sialidase. It is an unique enzyme, capable of masking the parasite’s presence from the host, while at the same time dampening the activation of CD8+ T cells, the most important components of the immune response. Since the description of Chagas’ disease in 1909, extensive research has identified important events in the disease in order to understand the biochemical mechanism that modulatesT. cruzi-host cell interactions and the ability of the parasite to ensure its survival. The importance of thetrans-sialidase enzyme brought life to many studies for the design of diagnostic tests, drugs and vaccines. While many groups have been prolific, such efforts have encountered problems, among them: the fact that whileT. cruzihave many genes that are unique to the parasite, it relies on multiple copies of them and the difficulty in providing epitopes that result in effective and robust immune responses. In this review, we aim to convey the importance oftrans-sialidase as well as to provide a history, including the initial failures and the most promising successes in the chasing of a working vaccine for a disease that is endemic in many tropical countries, including Brazil.
Attitude Towards and Adoption of the Novel Learning Environment Among Undergraduate Medical Students During COVID-19
Authors: Mohr Sonja , Küfe Birgit , Rheingans Anke , Guse Jennifer
Published: October 2021

Higher education has changed significantly since the beginning of the COVID-19 pandemic. Medical education programs with extensive practical and interactive components faced major challenges to protect students, faculty and patients. In response to COVID-19, many medical schools worldwide shut down undergraduate medical education and converted most of their teaching to digital formats. The aim of this paper is to assess the attitude towards and adoption of the novel learning environment among undergraduate medical students during COVID-19. Two studies were conducted to analyze specific aspects of the adaption of digital teaching during the digital summer semester 2020 (study 1), and to compare student satisfaction and their feeling of preparedness for exams in the digital semester compared to traditional semesters (study 2). Results show that there are numerous pros and cons of digital teaching. Pros were the large flexibility and large-scale availability of digital teaching materials. Cons were the lack of interactions with peers, professionals, and patients in practice. Results also show that female students as well as year 1 students seemingly coped better with the digital learning environment. Students with childcare or job obligations also benefited from the large flexibility. While student satisfaction decreased in the digital semester, they did not feel less prepared for exams. Cross-sectional comparisons revealed that student satisfaction and agreement gradually decreased in the comparison of the different cohorts with fourth year students being the least satisfied and showing the least agreement regarding the feeling of preparedness for exams. Altogether, our results indicate that students were able to cope with digital teaching, but clearly, some groups of students were able to better adapt to the novel learning environment. This might demand the introduction of tailored educational support services for different groups of students during COVID-19 as they progress through medical school.
Development and Validation of a Ferroptosis-Related Gene Signature and Nomogram for Predicting the Prognosis of Esophageal Squamous Cell Carcinoma
Authors: Ye Jiecheng , Wu Yining , Cai Heyuan , Sun Li , Deng Wanying , Liang Ruikun , Han Anjia
Published: October 2021

Esophageal squamous cell carcinoma (ESCC) is a common malignant tumor with high mortality and poor prognosis. Ferroptosis is a newly discovered form of cell death induced by iron-catalyzed excessive peroxidation of polyunsaturated fatty acids (PUFAs). However, the prognostic value of ferroptosis-related genes (FRGs) for ESCC remains unclear. Based on the ESCC dataset from the Gene Expression Omnibus (GEO) database, we identified 39 prognostic FRGs through univariate Cox regression analysis. After LASSO regression and multivariate Cox regression analyses, a multigene signature based on 10 prognostic FRGs was constructed and successfully divided ESCC patients into two risk groups. Patients in the low-risk group showed a significantly better prognosis than patients in the high-risk group. In addition, we combined the risk score with clinical predictors to construct a nomogram for ESCC. The predictive ability of the nomogram was further verified by ROC curves and calibration plots in both the training and validation sets. The predictive power of the nomogram was demonstrated to be better than that of either the risk score or clinical variable alone. Furthermore, functional analysis revealed that the 10-FRG signature was mainly associated with ferroptosis, differentiation and immune response. Connectivity map analysis identified potential compounds capable of targeting FRGs in ESCC. Finally, we demonstrated the prognostic value of SRC gene in ESCC using the clinical samples and found that SRC inhibition sensitized ESCC cells to ferroptosis inducers by in vitro experiments. In conclusion, we identified and verified a 10-FRG prognostic signature and a nomogram, which provide individualized prognosis prediction and provide insight into potential therapeutic targets for ESCC.
Developing an Embedding, Koopman and Autoencoder Technologies-Based Multi-Omics Time Series Predictive Model (EKATP) for Systems Biology research
Authors: Liu Suran , You Yujie , Tong Zhaoqi , Zhang Le
Published: October 2021

It is very important for systems biologists to predict the state of the multi-omics time series for disease occurrence and health detection. However, it is difficult to make the prediction due to the high-dimensional, nonlinear and noisy characteristics of the multi-omics time series data. For this reason, this study innovatively proposes an Embedding, Koopman and Autoencoder technologies-based multi-omics time series predictive model (EKATP) to predict the future state of a high-dimensional nonlinear multi-omics time series. We evaluate this EKATP by using a genomics time series with chaotic behavior, a proteomics time series with oscillating behavior and a metabolomics time series with flow behavior. The computational experiments demonstrate that our proposed EKATP can substantially improve the accuracy, robustness and generalizability to predict the future state of a time series for multi-omics data.
Nucleotide Diversity of the Maize ZmCNR13 Gene and Association With Ear Traits
Authors: Zuo Zhihao , Lu Yue , Zhu Minyan , Chen Rujia , Zhang Enying , Hao Derong , Huang Qianfeng , Wang Hanyao , Su Yanze , Wang Zhichao , Xu Yang , Li Pengcheng , Xu Chenwu , Yang Zefeng
Published: October 2021

The maize (Zea mays L.) ZmCNR13 gene, encoding a protein of fw2.2-like (FWL) family, has been demonstrated to be involved in cell division, expansion, and differentiation. In the present study, the genomic sequences of the ZmCNR13 locus were re-sequenced in 224 inbred lines, 56 landraces and 30 teosintes, and the nucleotide polymorphism and selection signature were estimated. A total of 501 variants, including 415 SNPs and 86 Indels, were detected. Among them, 51 SNPs and 4 Indels were located in the coding regions. Although neutrality tests revealed that this locus had escaped from artificial selection during the process of maize domestication, the population of inbred lines possesses lower nucleotide diversity and decay of linkage disequilibrium. To estimate the association between sequence variants of ZmCNR13 and maize ear characteristics, a total of ten ear-related traits were obtained from the selected inbred lines. Four variants were found to be significantly associated with six ear-related traits. Among them, SNP2305, a non-synonymous mutation in exon 2, was found to be associated with ear weight, ear grain weight, ear diameter and ear row number, and explained 4.59, 4.61, 4.31, and 8.42% of the phenotypic variations, respectively. These results revealed that natural variations of ZmCNR13 might be involved in ear development and can be used in genetic improvement of maize ear-related traits.
Case Report: The Carotid Body in COVID-19: Histopathological and Virological Analyses of an Autopsy Case Series
Authors: Porzionato Andrea , Emmi Aron , Contran Martina , Stocco Elena , Riccetti Silvia , Sinigaglia Alessandro , Macchi Veronica , Barzon Luisa , De Caro Raffaele
Published: October 2021

Various authors have hypothesized carotid body (CB) involvement in Coronavirus Disease 2019 (COVID-19), through direct invasion or indirect effects by systemic stimuli (‘cytokine storm’, angiotensin-converting enzyme [ACE]1/ACE2 imbalance). However, empirical evidence is limited or partial. Here, we present an integrated histopathological and virological analysis of CBs sampled at autopsy from four subjects (2 males and 2 females; age: >70 years old) who died of COVID-19. Histopathological, immunohistochemical and molecular investigation techniques were employed to characterize Severe Acute Respiratory Syndrome – Coronavirus 2 (SARS-CoV2) viral invasion and inflammatory reaction. SARS-CoV2 RNA was detected in the CBs of three cases through Real-Time Reverse Transcription Polymerase Chain Reaction (RT-PCR). In these cases, positive immunostaining for Nucleocapsid and Spike protein were also demonstrated, mainly at the level of large roundish cells consistent with type I cells, confirming direct CB invasion. In these cases, T lymphocytes showed focal aggregations in the CBs, suggestive of local inflammatory reaction. Blood congestion and microthrombosis were also found in one of the positive cases. Intriguingly, microthrombosis, blood congestion and microhaemorrages were also bilaterally detected in the CBs of the negative case, supporting the possibility of COVID-19 effects on the CB even in the absence of its direct invasion. SARS-CoV-2 direct invasion of the CB is confirmed through both immunohistochemistry and RT-PCR, with likely involvement of different cell types. We also reported histopathological findings which could be ascribed to local and/or systemic actions of SARS-CoV-2 and which could potentially affect chemoreception.
Preventive Administration of Non-Allergenic Bet v 1 Peptides Reduces Allergic Sensitization to Major Birch Pollen Allergen, Bet v 1
Authors: Akinfenwa Oluwatoyin , Huang Huey-Jy , Linhart Birgit , Focke-Tejkl Margarete , Vrtala Susanne , Poroshina Alina , Nikonova Alexandra , Khaitov Musa , Campion Nicholas J. , Eckl-Dorna Julia , Niederberger-Leppin Verena , Kratzer Bernhard , Tauber Peter Anton , Pickl Winfried F. , Kundi Michael , Campana Raffaela , Valenta Rudolf
Published: October 2021

IgE-mediated allergy to birch pollen affects more than 100 million patients world-wide. Bet v 1, a 17 kDa protein is the major allergen in birch pollen responsible for allergic rhinoconjunctivitis and asthma in birch pollen allergic patients. Allergen-specific immunotherapy (AIT) based on therapeutic administration of Bet v 1-containing vaccines is an effective treatment for birch pollen allergy but no allergen-specific forms of prevention are available. We developed a mouse model for IgE sensitization to Bet v 1 based on subcutaneous injection of aluminum-hydroxide adsorbed recombinant Bet v 1 and performed a detailed characterization of the specificities of the IgE, IgG and CD4+T cell responses in sensitized mice using seven synthetic peptides of 31-42 amino acids length which comprised the Bet v 1 sequence and the epitopes recognized by human CD4+T cells. We then demonstrate that preventive systemic administration of a mix of synthetic non-allergenic Bet v 1 peptides to 3-4 week old mice significantly reduced allergic immune responses, including IgE, IgG, IgE-mediated basophil activation, CD4+T cell and IL-4 responses to the complete Bet v 1 allergen but not to the unrelated major grass pollen allergen Phl p 5, without inducing Bet v 1-specific allergic sensitization or adaptive immunity. Our results thus demonstrate that early preventive administration of non-allergenic synthetic T cell epitope-containing allergen peptides could be a safe strategy for the prevention of allergen-specific IgE sensitization.
Processing and Preparation Method for High-Quality Opto-Electronic Perovskite Film
Authors: Chen Zheng , He Ping , Wu Dan , Chen Chen , Mujahid Muhammad , Li Ye , Duan Yu
Published: October 2021

The key to improving the energy conversion efficiency of perovskite solar cells lies in the optimization of the film morphology. The optical and electrical properties of the perovskite film, such as light absorption, carrier diffusion length, and charge transport, are all directly affected by the film morphology. Therefore, this review starts from the perovskite solar cells structure, and it summarizes the state-of-art perovskite film fabrication technologies and the caused film morphology to the performance perovskite solar cells. The spin coating method has an enormous waste of materials and only a small area of the device can be utilized. It is difficult to be used in commercial manufacturing. However, due to the high efficiency of this preparation method, it is irreplaceable in the initial research and development of perovskite materials, and so this method will be popular for a long time in the laboratory. Chemical vapor deposition and thermal vapor deposition have high technical requirements and a good repeatability of processing and manufacturing, and large-scale production can be realized. It may be the first technology to admit industrial application; the scratch coating method and slot-die have significant technical aspects. The similarity of the roll-to-roll manufacturing technology is also an efficient preparation method. Still, to achieve high-efficiency devices, it is necessary to consider the thickness control of each functional layer, and to find or prepare perovskite paste. Finally, we summarized the various fabrication processes and the prospects for the commercialization of perovskite solar cells. We predict that to achieve the commercialization of perovskite solar cells, the existing fabrication technologies should be optimized and more studies should be conducted.
An Eight Year Experience of Autologous Oocyte Vitrification for Infertile Patients Owing to Unavailability of Sperm on Oocyte Retrieval Day
Authors: Fu Xiao , Liu Xiaojie , Li Jing , Zhang Meng , Jiang Jingjing , Chen Qianqian , Li Mei , Gao Shanshan , Ma Jinlong
Published: October 2021

Objective: The objective of this study was to provide a descriptive analysis of the clinical outcomes achieved in oocyte vitrification in cases where sperm was unavailable on oocyte retrieval day, and to identify predictors of oocyte survival.

Methods: This retrospective cohort study used data from a university-affiliated reproductive medical center. There were 321 cycles in which some of, or all oocytes were vitrified owing to the unavailability of sperm between March 2009 and October 2017. A descriptive analysis of the clinical outcomes including both fresh embryo transfers and cryopreserved embryo transfers was provided. The ability of an individual parameter to forecast oocyte survival per thawing cycle was assessed by binary logistic regression analysis. The cumulative probability of live birth (CPLB) was estimated by using the Kaplan-Meier method according to the total number of oocytes thawed in consecutive procedures.

Results: The average survival rate was 83.13%. High-quality embryo rate and blastocyst rate decreased significantly decreased significantly in vitrification oocyte group compared to fresh control oocytes. The comparison of sibling oocytes in part-oocyte-vitrified cycles shows fewer high-quality embryos developed in the vitrified group. The live birth rate per warmed-oocyte was 4.3%. Reasons for lack of sperm availability on oocyte retrieval day and serum cholesterol levels were found to be associated with oocyte survival rate in the present study. Kaplan-Meier analysis showed no significant difference in CPLB between patients ≤35 vs. >35 years.

Conclusions: Oocyte vitrification is an indispensable and effective alternative when sperm are not available on oocyte retrieval day. The present study provided evidence that oocytes from infertile couples were more likely to suffer oocyte/embryo vitrification injury. Clinicians need to take this into account when advising patients in similar situations. Further studies will be necessary to clarify the correlation between serum metabolism parameters and human oocyte survival after vitrification.
Effects of Arbuscular Mycorrhizal Fungi on Rice Growth Under Different Flooding and Shading Regimes
Authors: Wang Yutao , Bao Xiaozhe , Li Shaoshan
Published: October 2021

Arbuscular mycorrhizal fungi (AMF) are present in paddy fields, where they suffer from periodic soil flooding and sometimes shading stress, but their interaction with rice plants in these environments is not yet fully explained. Based on two greenhouse experiments, we examined rice-growth response to AMF under different flooding and/or shading regimes to survey the regulatory effects of flooding on the mycorrhizal responses of rice plants under different light conditions. AMF had positive or neutral effects on the growth and yields of both tested rice varieties under non-flooding conditions but suppressed them under all flooding and/or shading regimes, emphasizing the high importance of flooding and shading conditions in determining the mycorrhizal effects. Further analyses indicated that flooding and shading both reduced the AMF colonization and extraradical hyphal density (EHD), implying a possible reduction of carbon investment from rice to AMF. The expression profiles of mycorrhizal P pathway marker genes (GintPT and OsPT11) suggested the P delivery from AMF to rice roots under all flooding and shading conditions. Nevertheless, flooding and shading both decreased the mycorrhizal P benefit of rice plants, as indicated by the significant decrease of mycorrhizal P responses (MPRs), contributing to the negative mycorrhizal effects on rice production. The expression profiles of rice defense marker genes OsPR1 and OsPBZ1 suggested that regardless of mycorrhizal growth responses (MGRs), AMF colonization triggered the basal defense response, especially under shading conditions, implying the multifaceted functions of AMF symbiosis and their effects on rice performance. In conclusion, this study found that flooding and shading both modulated the outcome of AMF symbiosis for rice plants, partially by influencing the mycorrhizal P benefit. This finding has important implications for AMF application in rice production.
Integrating Whole Blood Transcriptomic Collection Procedures Into the Current Anti-Doping Testing System, Including Long-Term Storage and Re-Testing of Anti-Doping Samples
Authors: Lima Giscard , Kolliari-Turner Alexander , Malinsky Fernanda Rossell , Guppy Fergus M. , Martin Renan Paulo , Wang Guan , Voss Sven Christian , Georgakopoulos Costas , Borrione Paolo , Pigozzi Fabio , Pitsiladis Yannis
Published: October 2021

Introduction: Recombinant human erythropoietin (rHuEPO) administration studies involving transcriptomic approaches have demonstrated a gene expression signature that could aid blood doping detection. However, current anti-doping testing does not involve collecting whole blood into tubes with RNA preservative. This study investigated if whole blood in long-term storage and whole blood left over from standard hematological testing in short-term storage could be used for transcriptomic analysis despite lacking RNA preservation.

Methods: Whole blood samples were collected from twelve and fourteen healthy nonathletic males, for long-term and short-term storage experiments. Long-term storage involved whole blood collected into Tempus™ tubes and K2EDTA tubes and subjected to long-term (i.e., ‒80°C) storage and RNA extracted. Short-term storage involved whole blood collected into K2EDTA tubes and stored at 4°C for 6‒48 h and then incubated at room temperature for 1 and 2 h prior to addition of RNA preservative. RNA quantity, purity, and integrity were analyzed in addition to RNA-Seq using the MGI DNBSEQ-G400 on RNA from both the short- and long-term storage studies. Genes presenting a fold change (FC) of >1.1 or < ‒1.1 with p ≤ 0.05 for each comparison were considered differentially expressed. Microarray analysis using the Affymetrix GeneChip® Human Transcriptome 2.0 Array was additionally conducted on RNA from the short-term study with a false discovery ratio (FDR) of ≤0.05 and an FC of >1.1 or < ‒1.1 applied to identify differentially expressed genes.

Results: RNA quantity, purity, and integrity from whole blood subjected to short- and long-term storage were sufficient for gene expression analysis. Long-term storage: when comparing blood tubes with and without RNA preservation 4,058 transcripts (6% of coding and non-coding transcripts) were differentially expressed using microarray and 658 genes (3.4% of mapped genes) were differentially expressed using RNA-Seq. Short-term storage: mean RNA integrity and yield were not significantly different at any of the time points. RNA-Seq analysis revealed a very small number of differentially expressed genes (70 or 1.37% of mapped genes) when comparing samples stored between 6 and 48 h without RNA preservative. None of the genes previously identified in rHuEPO administration studies were differently expressed in either long- or short-term storage experiments.

Conclusion: RNA quantity, purity, and integrity were not significantly compromised from short- or long-term storage in blood storage tubes lacking RNA stabilization, indicating that transcriptomic analysis could be conducted using anti-doping samples collected or biobanked without RNA preservation.
Coupling Between Production of Ribosomal RNA and Maturation: Just at the Beginning
Authors: Azouzi Chaima , Jaafar Mariam , Dez Christophe , Abou Merhi Raghida , Lesne Annick , Henras Anthony K. , Gadal Olivier
Published: October 2021

Ribosomal RNA (rRNA) production represents the most active transcription in the cell. Synthesis of the large rRNA precursors (35S/47S in yeast/human) is achieved by up to hundreds of RNA polymerase I (Pol I) enzymes simultaneously transcribing a single rRNA gene. In this review, we present recent advances in understanding the coupling between rRNA production and nascent rRNA folding. Mapping of the distribution of Pol I along ribosomal DNA at nucleotide resolution, using either native elongating transcript sequencing (NET-Seq) or crosslinking and analysis of cDNAs (CRAC), revealed frequent Pol I pausing, and CRAC results revealed a direct coupling between pausing and nascent RNA folding. High density of Pol I per gene imposes topological constraints that establish a defined pattern of polymerase distribution along the gene, with a persistent spacing between transcribing enzymes. RNA folding during transcription directly acts as an anti-pausing mechanism, implying that proper folding of the nascent rRNA favors elongation in vivo. Defects in co-transcriptional folding of rRNA are likely to induce Pol I pausing. We propose that premature termination of transcription, at defined positions, can control rRNA production in vivo.
C-Reactive Protein Levels and Gadolinium-Enhancing Lesions Are Associated With the Degree of Depressive Symptoms in Newly Diagnosed Multiple Sclerosis
Authors: Yalachkov Yavor , Anschuetz Victoria , Jakob Jasmin , Schaller-Paule Martin A. , Schaefer Jan Hendrik , Reilaender Annemarie , Friedauer Lucie , Behrens Marion , Foerch Christian
Published: October 2021

Background: Inflammation is essential for the pathogenesis of multiple sclerosis (MS). While the immune system contribution to the development of neurological symptoms has been intensively studied, inflammatory biomarkers for mental symptoms such as depression are poorly understood in the context of MS. Here, we test if depression correlates with peripheral and central inflammation markers in MS patients as soon as the diagnosis is established.

Methods: Forty-four patients were newly diagnosed with relapsing-remitting MS, primary progressive MS or clinically isolated syndrome. Age, gender, EDSS, C-reactive protein (CRP), albumin, white blood cells count in cerebrospinal fluid (CSF WBC), presence of gadolinium enhanced lesions (GE) on T1-weighted images and total number of typical MS lesion locations were included in linear regression models to predict Beck Depression Inventory (BDI) score and the depression dimension of the Symptoms Checklist 90-Revised (SCL90RD).

Results: CRP elevation and GE predicted significantly BDI (CRP: p = 0.007; GE: p = 0.019) and SCL90RD (CRP: p = 0.004; GE: p = 0.049). The combination of both factors resulted in more pronounced depressive symptoms (p = 0.04). CSF WBC and EDSS as well as the other variables were not correlated with depressive symptoms.

Conclusions: CRP elevation and GE are associated with depressive symptoms in newly diagnosed MS patients. These markers can be used to identify MS patients exhibiting a high risk for the development of depressive symptoms in early phases of the disease.
Association of Decreased Serum BDNF With Restless Legs Syndrome in Parkinson's Disease Patients
Authors: Huang Yi-xian , Zhang Qi-lin , Huang Cai-li , Wu Wen-qi , Sun Jia-wei
Published: October 2021

Objective: To objective of the study was to investigate whether serum brain-derived neurotrophic factor (BDNF) levels are associated with the severity of restless legs syndrome (RLS) in Parkinson's disease (PD).

Methods: A total of 249 PD patients with (n = 53) and without RLS (n = 196) and 326 age-matched controls were included in this study. All the serum BDNF levels of the participants were measured. The International Restless Legs Syndrome Study Group Rating Scale (IRLSSG-RS) was administered for the severity of RLS. The severity of PD patients were assessed by the Unified PD Rating Scale (UPDRS) and the Hoehn and Yahr (H-Y) stage.

Results: The prevalence of RLS was significantly higher in PD patients (21.3%) than in the controls group (7.4%) (p < 0.05). The IRLSSG-RS score in PD patients with RLS (16.25 ± 5.24) was significantly increased than in controls with RLS (12.08 ± 3.99) (p < 0.01). The serum BDNF levels were significantly decreased in PD patients with RLS than in PD patients without RLS, controls without RLS, and controls with RLS (p < 0.001). BDNF levels were negatively associated with IRLSSG-RS in both PD patients with RLS and controls with RLS group (both p < 0.01). Multiple regression analysis confirmed that in either PD with RLS or controls with RLS group, BDNF was an independent contributor to IRLSSG-RS (both p < 0.01).

Conclusions: Decreased serum BDNF levels may be involved in the pathophysiology of RLS in PD, suggesting that it may serve as a potential blood biomarker of diagnostic value for RLS in PD.
Development of the First WHO Guidelines for Risk Reduction of Cognitive Decline and Dementia: Lessons Learned and Future Directions
Authors: Stephen Ruth , Barbera Mariagnese , Peters Ruth , Ee Nicole , Zheng Lidan , Lehtisalo Jenni , Kulmala Jenni , Håkansson Krister , Chowdhary Neerja , Dua Tarun , Solomon Alina , Anstey Kaarin J. , Kivipelto Miia
Published: October 2021

The first WHO guidelines for risk reduction of cognitive decline and dementia marked an important milestone in the field of dementia prevention. In this paper, we discuss the evidence reviewed as part of the guidelines development and present the main themes emerged from its synthesis, to inform future research and policies on dementia risk reduction. The role of intervention effect-size; the mismatch between observational and intervention-based evidence; the heterogeneity of evidence among intervention trials; the importance of intervention duration; the role of timing of exposure to a certain risk factor and interventions; the relationship between intervention intensity and response; the link between individual risk factors and specific dementia pathologies; and the need for tailored interventions emerged as the main themes. The interaction and clustering of individual risk factors, including genetics, was identified as the overarching theme. The evidence collected indicates that multidomain approaches targeting simultaneously multiple risk factors and tailored at both individual and population level, are likely to be most effective and feasible in dementia risk reduction. The current status of multidomain intervention trials aimed to cognitive impairment/dementia prevention was also briefly reviewed. Primary results were presented focusing on methodological differences and the potential of design harmonization for improving evidence quality. Since multidomain intervention trials address a condition with slow clinical manifestation—like dementia—in a relatively short time frame, the need for surrogate outcomes was also discussed, with a specific focus on the potential utility of dementia risk scores. Finally, we considered how multidomain intervention could be most effectively implemented in a public health context and the implications world-wide for other non-communicable diseases targeting common risk factors, taking into account the limited evidence in low-middle income countries. In conclusion, the evidence from the first WHO guidelines for risk reduction of cognitive decline and dementia indicated that “one size does not fit all,” and multidomain approaches adaptable to different populations and individuals are likely to be the most effective. Harmonization in trial design, the use of appropriate outcome measures, and sustainability in large at-risk populations in the context of other chronic disorders also emerged as key elements.
LncRNA FOXP4-AS1 Promotes Progression of Ewing Sarcoma and Is Associated With Immune Infiltrates
Authors: Xiong Jiachao , Wu Liang , Huang Lu , Wu Chunyang , Liu Zhiming , Deng Wenqiang , Ma Shengbiao , Zhou Zhenhai , Yu Honggui , Cao Kai
Published: October 2021

Ewing sarcoma (ES) is a highly malignant primary bone tumor with poor prognosis. Studies have shown that abnormal expression of lncRNA influences the prognosis of tumor patients. Herein, we established that FOXP4-AS1 was up-regulated in ES and this correlated with poor prognosis. Further analysis illustrated that FOXP4-AS1 down-regulation repression growth, migration, along with invasion of ES. On the contrary, up-regulation of FOXP4-AS1 promoted the growth, migration, as well as invasion of ES. To explore the mechanism of FOXP4-AS1, Spearman correlation analysis was carried out to determine genes that were remarkably linked to FOXP4-AS1 expression. The potential functions and pathways involving FOXP4-AS1 were identified by GO analysis, Hallmark gene set enrichment analysis, GSEA, and GSVA. The subcellular fractionation results illustrated that FOXP4-AS1 was primarily located in the cytoplasm of ES cells. Then a ceRNA network of FOXP4-AS1 was constructed. Analysis of the ceRNA network and GSEA yielded two candidate mRNAs for FOXP4-AS1. Results of the combined survival analysis led us to speculate that FOXP4-AS1 may affect the expression of TMPO by sponging miR-298, thereby regulating the malignant phenotype of ES. Finally, we found that FOXP4-AS1 may modulates the tumor immune microenvironment in an extracellular vesicle-mediated manner. In summary, FOXP4-AS1 correlates with poor prognosis of ES. It promotes the growth, migration, as well as invasion of ES cells and may modulate the tumor immune microenvironment.
Suppression of EGFR/PKC-δ/NF-κB Signaling Associated With Imipramine-Inhibited Progression of Non-Small Cell Lung Cancer
Authors: Yueh Po-Fu , Lee Yuan-Hao , Chiang I-Tsang , Chen Wei-Ting , Lan Keng-Li , Chen Cheng-Hsien , Hsu Fei-Ting
Published: October 2021

Background

Anti-depressants have been reported to own anti-tumor potential types of cancers; however, the role of imipramine in non-small cell lung cancer (NSCLC) has not been elucidated. Epidermal growth factor receptor (EGFR) was known to be one of the key regulators that control NSCLC progression. Whether EGFR would be the target of imipramine for suppressing tumor signaling transduction and results in anti-tumor potential is remaining unclear.

Methods

We used CL-1-5-F4 cells and animal models to identify the underlying mechanism and therapeutic efficacy of imipramine. Cytotoxicity, apoptosis, invasion/migration, DNA damage, nuclear translocation of NF-κB, activation of NF-κB, phosphorylation of EGFR/PKC-δ/NF-κB was assayed by MTT, flow cytometry, transwell, wound healing assay, comet assay, immunofluorescence staining, NF-κB reporter gene assay and Western blotting, respectively. Tumor growth was validated by CL-1-5-F4/NF-κB-luc2 bearing animal model.

Results

Imipramine effectively induces apoptosis of NSCLC cells via both intrinsic and extrinsic apoptosis signaling. DNA damage was increased, while, invasion and migration potential of NSCLC cells was suppressed by imipramine. The phosphorylation of EGFR/PKC-δ/NF-κB and their downstream proteins were all decreased by imipramine. Similar tumor growth inhibition was found in imipramine with standard therapy erlotinib (EGFR inhibitor). Non-obvious body weight loss and liver pathology change were found in imipramine treatment mice.

Conclusion

Imipramine-triggered anti-NSCLC effects in both in vitro and in vivo model are at least partially attributed to its suppression of EGFR/PKC-δ/NF-κB pathway.
Case Report: An Internal Mammary Rhabdomyosarcoma After Mastectomy and Systemic and Radiation Therapy in a Patient With Breast Cancer
Authors: Wang Dan-Qiong , Zhang Jing-Yi , Li Jing , Ying Jian-Ming , Wang Xiang , Fan Ying , Wang Shu-Lian
Published: October 2021

Post-radiation soft tissue sarcomas (PRSTSs) are rare secondary malignancies. In this report, we describe the clinical presentation of a 52-year-old woman who underwent postmastectomy radiation therapy (PMRT) for left-sided breast cancer 2.7 years ago and presented with a left internal mammary mass and left interpectoral nodule on computed tomography. On further evaluation, she was diagnosed with internal mammary rhabdomyosarcoma and interpectoral nodal breast cancer relapse, and was treated with chemotherapy, followed by surgery and endocrine therapy. She developed left pleural metastases and is currently receiving targeted therapy. Internal mammary rhabdomyosarcomas are rare among PRSTSs and pose a diagnostic challenge for patients with breast cancer. Histological evaluation is important for the differential diagnosis of breast cancer relapses with secondary malignancies. The management of post-radiation thoracic rhabdomyosarcomas is challenging, and the prognosis is poor.
The Clinical Spectrum of Inflammatory Bowel Disease Associated With Specific Genetic Syndromes: Two Novel Pediatric Cases and a Systematic Review
Authors: Gatti Simona , Gelzoni Giulia , Catassi Giulia N. , Catassi Carlo
Published: October 2021

Background and Aims:Inflammatory bowel disease (IBD) is a typical polygenic disorder and less frequently shows a monogenic origin. Furthermore, IBD can originate in the context of specific genetic syndromes associated with a risk of autoimmune disorders. We aimed to systematically evaluate the prevalence of IBD in specific genetic syndromes and to review the clinical characteristics of the published cases.

Methods:According to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, studies describing patients with IBD and a genetic syndrome and/or studies indicating the prevalence or incidence of IBD in subjects with a genetic syndrome were included.

Results:Forty-six studies describing a total of 67 cases of IBD in six genetic syndromes and two personally assessed unpublished cases were included in the review. The majority of cases were associated with Turner syndrome (TS) (38 cases), Down syndrome (DS) (18 cases) and neurofibromatosis type 1 (NF1) (8 cases). Sporadic cases were described in DiGeorge syndrome (2), Kabuki syndrome (2), and Williams syndrome (1). The prevalence of IBD ranged from 0.67 to 4% in TS and from 0.2 to 1.57% in DS. The incidence of IBD was increased in TS and DS compared to the general population. Eight cases of IBD in TS had a severe/lethal course, many of which described before the year 2000. Two IBD cases in DS were particularly severe.

Conclusion:Evidence of a greater prevalence of IBD is accumulating in TS, DS, and NF1. Management of IBD in patients with these genetic conditions should consider the presence of comorbidities and possible drug toxicities.

Systematic Review Registration: PROSPERO, identifier: CRD42021249820
Structure–Activity Relationship of Novel Second-Generation Synthetic Cathinones: Mechanism of Action, Locomotion, Reward, and Immediate-Early Genes
Authors: Nadal-Gratacós Nuria , Alberto-Silva Ana Sofia , Rodríguez-Soler Míriam , Urquizu Edurne , Espinosa-Velasco Maria , Jäntsch Kathrin , Holy Marion , Batllori Xavier , Berzosa Xavier , Pubill David , Camarasa Jordi , Sitte Harald H. , Escubedo Elena , López-Arnau Raúl
Published: October 2021

Several new synthetic cathinones, which mimic the effect of classical psychostimulants such as cocaine or MDMA, have appeared in the global illicit drug market in the last decades. In fact, the illicit drug market is continually evolving by constantly adding small modifications to the common chemical structure of synthetic cathinones. Thus, the aim of this study was to investigate the in vitro and in vivo structure–activity relationship (SAR) of six novel synthetic cathinones currently popular as recreational drugs, pentedrone, pentylone, N-ethyl-pentedrone (NEPD), N-ethyl-pentylone (NEP), 4-methyl-pentedrone (4-MPD), and 4-methyl-ethylaminopentedrone (4-MeAP), which structurally differ in the absence or presence of different aromatic substituents and in their amino terminal group. Human embryonic kidney (HEK293) cells expressing the human isoforms of SERT and DAT were used for the uptake inhibition and release assays. Moreover, Swiss CD-1 mice were used to investigate the psychostimulant effect, rewarding properties (3, 10, and 30 mg/kg, i.p.), and the induction of immediate-early genes (IEGs), such as Arc and c-fos in the dorsal striatum (DS) and ventral striatum (VS) as well as bdnf in the medial prefrontal cortex (mPFC), of the test compounds. Our results demonstrated that all tested synthetic cathinones are potent dopamine (DA) uptake inhibitors, especially the N-ethyl analogs, while the ring-substituted cathinones tested showed higher potency as SERT inhibitors than their no ring-substituted analogs. Moreover, unlike NEP, the remaining test compounds showed clear “hybrid” properties, acting as DAT blockers but SERT substrates. Regarding the locomotion, NEP and NEPD were more efficacious (10 mg/kg) than their N-methyl analogs, which correlates with their higher potency inhibiting the DAT and an overexpression of Arc levels in the DS and VS. Furthermore, all compounds tested induced an increase in c-fos expression in the DS, except for 4-MPD, the least effective compound in inducing hyperlocomotion. Moreover, NEP induced an up-regulation of bdnf in the mPFC that correlates with its 5-HTergic properties. Finally, the present study demonstrated for the first time that NEP, 4-MPD, and 4-MeAP induce reward in mice. Altogether, this study provides valuable information about the mechanism of action and psychostimulant and rewarding properties as well as changes in the expression of IEGs related to addiction induced by novel second-generation synthetic cathinones.