bluuebunny/crossref_metadata_2025 huggingface dataset:
A comparative study on the tax and fee reduction policies for sustainable development: Empirical analysis from the world’s three major economies in tackling COVID-19
Authors: Yujuan Ma , Guangli Yang , Bo Li
Published: October 2022

The global COVID-19 epidemic in 2020 has caused a significant drop in many countries’ employment rates and economies. Numerous countries have implemented a number of tax and fee reduction policies in response to the COVID-19 epidemic. This essay analyzes the tax and fee policies and results of the three largest economies in the world—China, the United States, and Japan—in response to COVID-19 between 2020 and 2021. We will first compare the effects of tax and fee reduction policies horizontally across China, the US, and Japan. The second step is to compare the effects of tax and fee reduction policies. According to the study, China, the United States and Japan have each implemented separate tax and fee policies in response to COVID-19. The United States primarily provides corporate income tax, personal income tax relief, and increased social welfare and subsidies that reduce the tax burden of enterprises, stimulate the vitality of enterprises, increase the disposable income of residents, stimulate consumption, and thus promote economic growth; Japan primarily provides income tax, consumption tax and business tax relief to reduce the cost of consumption for residents, increase corporate profits and cash flow, restore economic growth and stabilize employment. China primarily provides tax relief for VAT, corporate income tax and social security funds. It indirectly lowers the selling price of products, reduces the operating costs of enterprises, increases personal disposable income, stimulates consumption, promotes the growth of business performance and plays a certain role in curbing inflation. Based on the empirical analysis of the impact of different tax and fee reduction policies on the economy and employment in China, the United States and Japan, it is of some relevance for other countries. Therefore, in response to COVID-19 or major emergencies, each country can formulate targeted tax and fee reduction measures based on its own national conditions to support economic growth and stable employment.
Environmental sustainability with the role of green innovation and economic growth in India with bootstrap ARDL approach
Authors: Pachiyappan Duraisamy , Alam Md Shabbir , Khan Uzma , Khan Aarif Mohammed , Mohammed Shariq , Alagirisamy Kuppusamy , Manigandan Palanisamy
Published: October 2022

This paper applies a novel Bootstrap Autoregressive Distributed Lag (BARDL) approach to investigate the relationship between green innovation (GI), economic growth (GDP), drama and film (D&F) industry, and environmental sustainability in India for the 1995 to 2020 period. The data has been checked for its stationarity by applying the Zivot and Andrews (ZA) unit root test, and the cointegration test results suggest a long-run equilibrium relationship between the variables. The empirical finding of long-run estimates reveals that 1% augments of GI, GDP, and D&F industry increase CO2 emissions by −0.079, 0.566%, and 0.143%, respectively. Furthermore, the main results indicate that GDP and the D&F industry have statistically significant positive effects on CO2 emissions, and GI has statistically significant negative effects on CO2 emissions. The GI leads to lower environmental damage by reducing carbon emissions. Regarding causal relationships, bidirectional causality is found between D&F and CO2 emissions, GI and CO2 emissions. In addition, a unidirectional causality is also revealed from GDP to CO2 emissions. Based on the finding of this study, policy implications are suggested for India.
Genetic deletions and high diversity of Plasmodium falciparum histidine-rich proteins 2 and 3 genes in parasite populations in Ghana
Authors: Duah-Quashie Nancy Odurowah , Opoku-Agyeman Philip , Bruku Selassie , Adams Tryphena , Tandoh Kwesi Zandoh , Ennuson Nana Aba , Matrevi Sena Adzoa , Abuaku Benjamin , Quashie Neils Ben , Watters Chaselynn , Wolfe David , Quijada Hugo Miranda , Sanders Terrel
Published: October 2022

Rapid diagnostic tests (RDTs) are used to diagnose malaria in Ghana and other malaria endemic countries. Plasmodium falciparum histidine-rich protein 2 (PFHRP2) based RDTs are widely used, however the occurrence of deletions of the pfhrp2 gene in some parasites have resulted in false negative test results. Monoclonal antibodies of PFHRP2 cross reacts with PFHRP3 because they share structural similarities and this complements the detection of the parasites by RDT. These two genes were investigated in Ghanaian P. falciparum parasite population to detect deletions and the polymorphisms in exon 2 of the pfhrp2 and pfhrp3 genes. Parasite isolates (2,540) from children ≤ 12 years with uncomplicated malaria from 2015 to 2020 transmission seasons were used. Both genes were amplified using nested PCR and negative results indicated the presence of the deletion of genes. Amplified genes were sequenced for the detection of the amino acid repeats. Deletions were observed in 30.7% (780/2,540) and 17.2% (438/2,540) of the samples for pfhrp2 and pfhrp3 respectively with increasing trends over the three time periods (χ2 −10.305, p = 0.001). A total of 1,632 amplicons were sequenced for each gene, analysis was done on 1,124 and 1,307 good quality sequences for pfhrp2 and pfhrp3 respectively. Pfhrp2 repeat polymorphisms were dominantly of types 2 (AHHAHHAAD) and 7 (AHHAAD) with large numbers of variants. A novel variant of type 14 (AHHANHATD) was seen for pfhrp2. For the pfhrp3 repeat types, 16 (AHHAAN), 17 (AHHDG) and 18 (AHHDD) were the dominant types observed. Variants of type 16 (AHHAAH) and (AHHASH) were also dominant. Repeat types 1, 2, 3, 4, 5, 6, 7, 8, 11, 13, 15, 16, and 19 were observed be shared by both genes. The haplotype diversity of both genes ranged between 0.872 and 1 indicating high diversity of the polymorphisms in the isolates. The implication of the findings of the frequencies of the pfhrp2 and pfhrp3 deletions as well as the variants of the main epitopes of the monoclonal antibodies for the RDT (types 2 and 7) in our isolates is an indication of decreased sensitivity of the RDTs in diagnosing malaria infections in Ghana.
Comprehensive analysis of epigenomics and transcriptome data to identify potential target genes associated with obesity
Authors: Wu Peili , Guo Lei , Li Xuelin , Du Yuejun , Lin Xiaochun , Ma Xiaoqin , Lin Yingbei , Wen Churan , Yang Chuyi , Liu Nannan , Feng Qijian , Xue Yaoming , Guan Meiping
Published: October 2022

DNA methylation is closely related to the occurrence and development of many diseases, but its role in obesity is still unclear. This study aimed to find the potential differentially methylated genes associated with obesity occurrence and development. By combining methylation and transcriptome analysis, we identified the key genes in adipose tissue affecting the occurrence and development of obesity and revealed the possible molecular mechanisms involved in obesity pathogenesis. We first screened 14 methylation-related differential genes and verified their expression in adipose tissue by quantitative polymerase chain reaction (qPCR). Seven genes with the same expression pattern were identified as key genes, namely, CCRL2, GPT, LGALS12, PC, SLC27A2, SLC4A4, and TTC36. Then, the immune microenvironment of adipose tissue was quantified by CIBERSORT, and we found that the content of M0 macrophages and T follicular helper cells in adipose tissue was significantly increased and decreased, respectively, in the obese group. Furthermore, the relationship between key genes and the immune microenvironment was analyzed. Additionally, the metabolic pathway activity of each sample was calculated based on the ssGSEA algorithm, and the key gene–metabolic network was constructed. Moreover, we performed a CMAP analysis based on the differential genes in adipose tissue to screen out drugs potentially effective in obesity treatment. In conclusion, we identified seven methylation-related key genes closely related to obesity pathogenesis and explored the potential mechanism of their role in obesity. This study provided novel insights into the molecular mechanisms and management of obesity.
Genetic evidence suggests that depression increases the risk of erectile dysfunction: A Mendelian randomization study
Authors: Ma Kai , Song Pan , Liu Zhenghuan , Yang Luchen , Wang Linchun , Zhou Jing , Chen Junhao , Dong Qiang
Published: October 2022

Background: The causal relationship between depression and erectile dysfunction (ED) is still uncertain.

Objectives: To identify the genetically predicted causality of depression on ED through Mendelian randomization (MR).

Materials and methods: A comprehensive GWAS meta-analysis comprising 807,553 Europeans provided single-nucleotide polymorphism (SNP) information for depression, and another genome-wide association analysis involving 223,805 European ancestries measured SNPs for ED. The inverse variance weighted (IVW) method was used as the primary MR analysis method to evaluate causal effects. In addition, the maximum likelihood method, MR-Egger, weighted median, robust adjusted contour score (MR.RAPS), and MR pleiotropic residual and outlier (MR-PRESSO) methods were used as supplements for sensitivity analysis.

Results: According to the IVW analysis, depression significantly increases the incidence of ED (odds ratio [OR] = 1.68, 95% confidence interval [CI] = 1.38–2.05, p < 0.001). In sensitivity analyses, the ORs for the maximum likelihood method, MR-Egger, weighted median, MR.RAPS, and MR-PRESSO are 1.70 (95% CI = 1.39–2.08, p < 0 .001), 1.94 (95% CI = 0.63–6.01, p > 0 .05), 1.59 (95% CI = 1.21–2.10, p < 0 .001), 1 .70 (95% CI = 1.39–2.08, p < 0 .001), and 1.68 (95% CI = 1.40–2.04, p < 0 .001). There is no clear indication of potential heterogeneity or pleiotropy (p for the MR-Egger intercept = 0.804; p for the global test = 0.594; and p for Cochran’s Q statistics >0.05).

Conclusion: Genetically predicted depression plays a potentially causal role in the occurrence of ED.
Establishment of m7G-related gene pair signature to predict overall survival in colorectal cancer
Authors: Li Kai , Wang Weixing
Published: October 2022

Background: N7-methylguanosine (m7G) is an emerging research hotspot in the field of RNA methylation, and its role in tumor regulation is becoming increasingly recognized. However, its role in colorectal cancer (CRC) remains unclear. Hence, our study explored the role of m7G in CRC.

Methods: The mRNA expression data and the corresponding clinical information of the patients with CRC were obtained from The Cancer Genome Atlas (TCGA). A m7G-related gene pair signature was established using the Cox and LASSO regression analyses. A series of in silico analyses based on the signature included analysis of prognosis, correlation analysis, immune-related analysis, and estimation of tumor mutational burden (TMB), microsatellite instability (MSI), and response to immunotherapy. A nomogram prediction model was then constructed.

Results: In total, 2156 m7G-related gene pairs were screened based on 152 m7G-related genes. Then, a prognostic signature of seven gene pairs was constructed, and the patients were stratified into high- or low-risk groups. Better overall survival (OS), left-sided tumor, early stage, immune activity, and low proportion of MSI-low and MSI-high were all associated with a low risk score. High-risk patients had a higher TMB, and patients with a high TMB had a poor OS. Furthermore, the risk score was linked to immune checkpoint expression (including PD-L1), the tumor immune dysfunction and exclusion (TIDE) score, and chemotherapy sensitivity. We also created an accurate nomogram to increase the clinical applicability of the risk score.

Conclusion: We identified an m7G pair-based prognostic signature associated with prognosis, immune landscape, immunotherapy, and chemotherapy in CRC. These findings could help us to better understand the role of m7G in CRC, as well as pave the path for novel methods to assess prognosis and design more effective individualized therapeutic strategies.
Gasdermin D protects against Streptococcus equi subsp. zooepidemicus infection through macrophage pyroptosis
Authors: Xu Guobin , Guo Zheng , Liu Yuxuan , Yang Yalin , Lin Yongjin , Li Chunliu , Huang Yunfei , Fu Qiang
Published: October 2022

Streptococcus equi subsp. zooepidemicus (S. zooepidemicus, SEZ) is an essential zoonotic bacterial pathogen that can cause various inflammation, such as meningitis, endocarditis, and pneumonia. Gasdermin D (GSDMD) is involved in cytokine release and cell death, indicating an important role in controlling the microbial infection. This study investigated the protective role of GSDMD in mice infected with SEZ and examined the role of GSDMD in peritoneal macrophages in the infection. GSDMD-deficient mice were more susceptible to intraperitoneal infection with SEZ, and the white pulp structure of the spleen was seriously damaged in GSDMD-deficient mice. Although the increased proportion of macrophages did not depend on GSDMD in both spleen and peritoneal lavage fluid (PLF), deficiency of GSDMD caused the minor release of the pro-inflammatory cytokines interleukin-1β (IL-1β) and interleukin-18 (IL-18) during the infection in vivo. In vitro, SEZ infection induced more release of IL-1β, IL-18, and lactate dehydrogenase (LDH) in wild-type macrophages than in GSDMD-deficient macrophages. Finally, we demonstrated that pore formation and pyroptosis of macrophages depended on GSDMD. Our findings highlight the host defense mechanisms of GSDMD against SEZ infection, providing a potential therapeutic target in SEZ infection.
IL-1β is not critical to chronic heart dysfunction in mice with Chagas disease
Authors: Sousa Oliveira Camila Victória , Moreno-Loaiza Oscar , Figueiredo-Vanzan Daniel , Peroba Ramos Isalira , Mata-Santos Hilton , Torres Bozza Marcelo , Neto Paiva Claudia , Medei Emiliano
Published: October 2022

Long after Trypanosoma cruzi infection, 40% of individuals develop a progressive chronic chagasic cardiomyopathy (CCC), with systolic dysfunction and arrhythmias. Since we previously showed IL-1β mediates the development of systolic dysfunction and cardiac arrhythmias in diabetes mellitus and cardiorenal syndrome, and IL-1β remains elevated in Chagas disease patients, here we tested the role of IL-1β in CCC using a mouse model. Mice deficient in IL-1R expression (Il-1r−/−) survived acute T. cruzi infection with greater parasitemia than controls but did not lose weight as wild-type (WT) did. At the chronic stage, WT presented prolonged ventricular repolarization intervals (QJ), while Il-1r−/− presented intervals like noninfected controls. Infected Il-1r−/− and WT did not differ in stroke volume (SV), the incidence of cardiac arrhythmias on electrocardiography (EKG), whole heart action potential duration (APD), or the incidence of triggered activity after S1–S2 protocol, which is a measure of susceptibility to cardiac arrhythmias. We also treated chronically infected WT mice with an IL-1R antagonist, anakinra. Treatment shortened the QJ interval but did not improve the SV or the incidence of cardiac arrhythmias on EKG. Anakinra failed to reduce triggered activity following the electrical extra-stimulation protocol. In conclusion, the absence of functional IL-1β/IL-1R signaling did not prevent or reverse the decrease of SV or the incidence of cardiac arrhythmias induced by chronic T. cruzi infection, implying this is not a critical mechanism in generating or maintaining CCC. Since similar cardiac abnormalities were previously credited to IL-1β signaling, ruling out this mechanism is important to discourage further attempts of IL-1β blockade as a therapeutical measure.
Increased prevalence of clonal hematopoiesis of indeterminate potential in hospitalized patients with COVID-19
Authors: Schenz Judith , Rump Katharina , Siegler Benedikt Hermann , Hemmerling Inga , Rahmel Tim , Thon Jan N. , Nowak Hartmuth , Fischer Dania , Hafner Anna , Tichy Lucas , Bomans Katharina , Meggendorfer Manja , Koos Björn , von Groote Thilo , Zarbock Alexander , Fiedler Mascha O. , Zemva Johanna , Larmann Jan , Merle Uta , Adamzik Michael , Müller-Tidow Carsten , Haferlach Torsten , Leuschner Florian , Weigand Markus A.
Published: October 2022

Clonal hematopoiesis of indeterminate potential (CHIP) leads to higher mortality, carries a cardiovascular risk and alters inflammation. All three aspects harbor overlaps with the clinical manifestation of COVID-19. This study aimed to identify the impact of CHIP on COVID-19 pathophysiology. 90 hospitalized patients were analyzed for CHIP. In addition, their disease course and outcome were evaluated. With a prevalence of 37.8%, the frequency of a CHIP-driver mutation was significantly higher than the prevalence expected based on median age (17%). CHIP increases the risk of hospitalization in the course of the disease but has no age-independent impact on the outcome within the group of hospitalized patients. Especially in younger patients (45 – 65 years), CHIP was associated with persistent lymphopenia. In older patients (> 65 years), on the other hand, CHIP-positive patients developed neutrophilia in the long run. To what extent increased values of cardiac biomarkers are caused by CHIP independent of age could not be elaborated solely based on this study. In conclusion, our results indicate an increased susceptibility to a severe course of COVID-19 requiring hospitalization associated with CHIP. Secondly, they link it to a differentially regulated cellular immune response under the pressure of SARS-CoV-2 infection. Hence, a patient’s CHIP-status bears the potential to serve as biomarker for risk stratification and to early guide treatment of COVID-19 patients.
Response of plankton community respiration under variable simulated upwelling events
Authors: Baños Isabel , Arístegui Javier , Benavides Mar , Gómez-Letona Markel , Montero María F. , Ortiz Joaquín , Schulz Kai G. , Ludwig Andrea , Riebesell Ulf
Published: October 2022

Climate change is expected to alter the intensity and frequency of upwelling in high productive coastal regions, thus impacting nutrient fluxes, primary productivity and consequently carbon cycling. However, it is unknown how these changes will impact the planktonic (phytoplankton and bacteria) community structure, which affects community respiration (CR) and hence the carbon available for sequestration or transfer to upper trophic levels. Here we present results from a 37-day mesocosm experiment where we examined the response of CR to nutrient additions by simulating upwelling events at different intensities (low, medium, high and extreme) and modes (singular and recurring additions). We also analysed the potential contribution of different plankton size classes and functional groups to CR. The trend in accumulated CR with respect to nutrient fertilisation (total nitrogen added during the experiment) was linear in the two modes. Microplankton (mostly diatoms) and nanoplankton (small flagellates) dominated under extreme upwelling intensities and high CR in both singular and recurring upwelling modes, explaining >65% of the observed variability in CR. In contrast, prokaryotic picoplankton (heterotrophic bacteria and autotrophic cyanobacteria) explained <43% of the variance in CR under the rest of the upwelling intensities and modes tested. Changes in planktonic community structure, while modulating CR variability, would regulate the metabolic balance of the ecosystem, shifting it towards net-heterotrophy when the community is dominated by small heterotrophs and to net-autotrophy when large autotrophs prevail; although depending on the mode in which nutrients are supplied to the system. This shift in the dominance of planktonic organism will hence affect not only CR but also carbon sequestration in upwelling regions
Play the plug: How bacteria modify recognition by host receptors?
Authors: Tiruvayipati Suma , Hameed Dharjath S. , Ahmed Niyaz
Published: October 2022

The diverse microbial community that colonizes the gastrointestinal tract has remarkable effects on the host immune system and physiology resulting in homeostasis or disease. In both scenarios, the gut microbiota interacts with their host through ligand-receptor binding whereby the downstream signaling processes determine the outcome of the interaction as disease or the counteractive immune responses of the host. Despite several studies on microbe-host interactions and the mechanisms by which this intricate process happens, a comprehensive and updated inventory of known ligand-receptor interactions and their roles in disease is paramount. The ligands which originate as a result of microbial responses to the host environment contribute to either symbiotic or parasitic relationships. On the other hand, the host receptors counteract the ligand actions by mounting a neutral or an innate response. The varying degrees of polymorphic changes in the host receptors contribute to specificity of interaction with the microbial ligands. Additionally, pathogenic microbes manipulate host receptors with endogenous enzymes belonging to the effector protein family. This review focuses on the diversity and similarity in the gut microbiome-host interactions both in health and disease conditions. It thus establishes an overview that can help identify potential therapeutic targets in response to critically soaring antimicrobial resistance as juxtaposed to tardy antibiotic development research.
Strain level and comprehensive microbiome analysis in inflammatory bowel disease via multi-technology meta-analysis identifies key bacterial influencers of disease
Authors: Ravichandar Jayamary Divya , Rutherford Erica , Chow Cheryl-Emiliane T. , Han Andrew , Yamamoto Mitsuko Lynn , Narayan Nicole , Kaplan Gilaad G. , Beck Paul L. , Claesson Marcus J. , Dabbagh Karim , Iwai Shoko , DeSantis Todd Z.
Published: October 2022

Objective

Inflammatory bowel disease (IBD) is a heterogenous disease in which the microbiome has been shown to play an important role. However, the precise homeostatic or pathological functions played by bacteria remain unclear. Most published studies report taxa-disease associations based on single-technology analysis of a single cohort, potentially biasing results to one clinical protocol, cohort, and molecular analysis technology. To begin to address this key question, precise identification of the bacteria implicated in IBD across cohorts is necessary.

Methods

We sought to take advantage of the numerous and diverse studies characterizing the microbiome in IBD to develop a multi-technology meta-analysis (MTMA) as a platform for aggregation of independently generated datasets, irrespective of DNA-profiling technique, in order to uncover the consistent microbial modulators of disease. We report the largest strain-level survey of IBD, integrating microbiome profiles from 3,407 samples from 21 datasets spanning 15 cohorts, three of which are presented for the first time in the current study, characterized using three DNA-profiling technologies, mapping all nucleotide data against known, culturable strain reference data.

Results

We identify several novel IBD associations with culturable strains that have so far remained elusive, including two genome-sequenced but uncharacterized Lachnospiraceae strains consistently decreased in both the gut luminal and mucosal contents of patients with IBD, and demonstrate that these strains are correlated with inflammation-related pathways that are known mechanisms targeted for treatment. Furthermore, comparative MTMA at the species versus strain level reveals that not all significant strain associations resulted in a corresponding species-level significance and conversely significant species associations are not always re-captured at the strain level.

Conclusion

We propose MTMA for uncovering experimentally testable strain-disease associations that, as demonstrated here, are beneficial in discovering mechanisms underpinning microbiome impact on disease or novel targets for therapeutic interventions.
Untargeted muscle tissue metabolites profiling in young, adult, and old rats supplemented with tocotrienol-rich fraction
Authors: Saud Gany Siti Liyana , Tan Jen Kit , Chin Kok Yong , Hakimi Nur Haleeda , Ab Rani Nazirah , Ihsan Nurhazirah , Makpol Suzana
Published: October 2022

The greatest significant influence on human life span and health is inevitable ageing. One of the distinguishing characteristics of ageing is the gradual decrease of muscle mass and physical function. There has been growing evidence that tocotrienol can guard against age-associated chronic diseases and metabolic disorders. This study aimed to elucidate the effects of tocotrienol-rich fraction (TRF) on muscle metabolomes and metabolic pathways in ageing Sprague Dawley (SD) rats. Three months, 9 months, and 21 months old male SD rats were divided into control and treated groups with 10 rats per group. Rats in control and treated groups were given 60 mg/kg body weight/day of palm olein and 60 mg/kg body weight/day of TRF, respectively, via oral gavage for 3 months. Muscle performance was assessed at 0 and 3 months of treatment by measuring muscle strength and function. Our results showed that TRF treatment caused a significant increase in the swimming time of the young rats. Comparison in the control groups showed that metabolites involved in lipid metabolisms such as L-palmitoyl carnitine and decanoyl carnitine were increased in ageing. In contrast, several metabolites, such as 3-phosphoglyceric acid, aspartic acid and aspartyl phenylalanine were decreased. These findings indicated that muscle metabolomes involved in lipid metabolism were upregulated in aged rats. In contrast, the metabolites involved in energy and amino acid metabolism were significantly downregulated. Comparison in the TRF-supplemented groups showed an upregulation of metabolites involved in energy and amino acid metabolism. Metabolites such as N6-methyl adenosine, spermine, phenylalanine, tryptophan, aspartic acid, histidine, and N-acetyl neuraminic acid were up-regulated, indicating promotion of amino acid synthesis and muscle regeneration. Energy metabolism was also improved in adult and old rats with TRF supplementation as indicated by the upregulation of nicotinamide adenine dinucleotide and glycerol 3-phosphate compared to the control group. In conclusion, the mechanism underlying the changes in skeletal muscle mass and functions in ageing was related to carbohydrate, lipid and amino acid metabolism. Tocotrienol supplementation showed beneficial effects in alleviating energy and amino acid synthesis that may promote the regeneration and renewal of skeletal muscle in ageing rats.
The endocannabinoidome mediator N-oleoylglycine is a novel protective agent against 1-methyl-4-phenyl-pyridinium-induced neurotoxicity
Authors: Lauritano Anna , Cipollone Irene , Verde Roberta , Kalkan Hilal , Moriello Claudia , Iannotti Fabio Arturo , Di Marzo Vincenzo , Piscitelli Fabiana
Published: October 2022

N-oleoylglycine (OlGly) is a lipid mediator that belongs to the expanded version of the endocannabinoid (eCB) system, the endocannabinoidome (eCBome), which has recently gained increasing attention from the scientific community for its protective effects in a mouse model of mild traumatic brain injury. However, the effects of OlGly on cellular models of Parkinson’s disease (PD) have not yet been investigated, whilst other lipoaminoacids have been reported to have beneficial effects. Moreover, the protective effects of OlGly seem to be mediated by direct activation of proliferator-activated receptor alpha (PPARα), which has already been investigated as a therapeutic target for PD. Therefore, this study aims to investigate the possible protective effects of OlGly in an in vitro model obtained by treating the neuroblastoma cell line, SH-SY5Y (both differentiated and not) with 1-methyl-4-phenyl-pyridinium (MPP+), which mimics some cellular aspects of a PD-like phenotype, in the presence or absence of the PPARα antagonist, GW6471. Our data show that MPP+ increases mRNA levels of PPARα in both non differentiated and differentiated cells. Using assays to assess cell metabolic activity, cell proliferation, and pro-inflammatory markers, we observed that OlGly (1 nM), both as treatment (1 h) and pre-treatment (4 h), is able to protect against neuronal damage induced by 24 h MPP+ exposure through PPARα. Moreover, using a targeted lipidomics approach, we demonstrate that OlGly exerts its effects also through the modulation of the eCBome. Finally, treatment with OlGly was able also to reduce increased IL-1β induced by MPP+ in differentiated cells. In conclusion, our results suggest that OlGly could be a promising therapeutic agent for the treatment of MPP+-induced neurotoxicity.
A combined use of silver pretreatment and impregnation with consequent Nissl staining for cortex and striatum architectonics study
Authors: Piavchenko Gennadii , Soldatov Vladislav , Venediktov Artem , Kartashkina Natalia , Novikova Natalia , Gorbunova Marina , Boronikhina Tatiana , Yatskovskiy Alexander , Meglinski Igor , Kuznetsov Sergey
Published: October 2022

Despite a rapid growth in the application of modern techniques for visualization studies in life sciences, the classical methods of histological examination are yet to be outdated. Herein, we introduce a new approach that involves combining silver nitrate pretreatment and impregnation with consequent Nissl (cresyl violet) staining for cortex and striatum architectonics study on the same microscopy slide. The developed approach of hybrid staining provides a high-quality visualization of cellular and subcellular structures, including impregnated neurons (about 10%), Nissl-stained neurons (all the remaining ones), and astrocytes, as well as chromatophilic substances, nucleoli, and neuropil in paraffin sections. We provide a comparative study of the neuronal architectonics in both the motor cortex and striatum based on the differences in their tinctorial properties. In addition to a comparative study of the neuronal architectonics in both the motor cortex and striatum, the traditional methods to stain the cortex (motor and piriform) and the striatum are considered. The proposed staining approach compiles the routine conventional methods for thin sections, expanding avenues for more advanced examination of neurons, blood–brain barrier components, and fibers both under normal and pathological conditions. One of the main hallmarks of our method is the ability to detect changes in the number of glial cells. The results of astrocyte visualization in the motor cortex obtained by the developed technique agree well with the alternative studies by glial fibrillary acidic protein (GFAP) immunohistochemical reaction. The presented approach of combined staining has great potential in current histological practice, in particular for the evaluation of several neurological disorders in clinical, pre-clinical, or neurobiological animal studies.
Psychopathic tendency in violent offenders is associated with reduced aversive Pavlovian inhibition of behavior and associated striatal BOLD signal
Authors: Geurts Dirk E. M. , von Borries Katinka , Huys Quentin J. M. , Bulten Berend H. , Verkes Robbert-Jan , Cools Roshan
Published: October 2022

Background

Violent offenders with psychopathic tendencies are characterized by instrumental, i.e., planned, callous, and unemotional (aggressive) behavior and have been shown to exhibit abnormal aversive processing. However, the consequences of abnormal aversive processing for instrumental action and associated neural mechanisms are unclear.

Materials and methods

Here we address this issue by using event-related functional magnetic resonance imaging (fMRI) in 15 violent offenders with high psychopathic tendencies and 18 matched controls during the performance of an aversive Pavlovian-to-instrumental transfer paradigm. This paradigm allowed us to assess the degree to which aversive Pavlovian cues affect instrumental action and associated neural signaling.

Results

Psychopathic tendency scores were associated with an attenuation of aversive Pavlovian inhibition of instrumental action. Moreover, exploratory analyses revealed an anomalous positive association between aversive inhibition of action and aversive inhibition of BOLD signal in the caudate nucleus of violent offenders with psychopathic tendencies. In addition, psychopathic tendency also correlated positively with amygdala reactivity during aversive versus neutral cues in Pavlovian training.

Conclusion

These findings strengthen the hypothesis that psychopathic tendencies in violent offenders are related to abnormal impact of aversive processing on instrumental behavior. The neural effects raise the possibility that this reflects deficient transfer of aversive Pavlovian inhibitory biases onto neural systems that implement instrumental action, including the caudate nucleus.
CCR7-mediated T follicular helper cell differentiation is associated with the pathogenesis and immune microenvironment of spinal cord injury-induced immune deficiency syndrome
Authors: Li Chaochen , Wu Chunshuai , Xu Guanhua , Liu Yang , Chen Jiajia , Zhang Jinlong , Hong Hongxiang , Ji Chunyan , Cui Zhiming
Published: October 2022

Spinal cord injury-induced immune deficiency syndrome (SCI-IDS) is a disorder characterized by systemic immunosuppression secondary to SCI that dramatically increases the likelihood of infection and is difficult to treat. T follicular helper (Tfh) cells regulated by chemokine receptor CCR7 are associated with SCI-IDS after acute SCI. The present study explored the roles of CCR7 in SCI-IDS occurrence and immune microenvironment composition. Gene expression profile data of peripheral blood leukocytes from SCI and non-SCI subjects were collected from the Gene Expression Omnibus database. According to differential gene expression analysis, a protein-protein interaction (PPI) network, and risk model construction, the CCR7 expression level was prominently related to acute SCI and CCR7 expression was significantly downregulated after acute SCI. Next, we constructed a clinical prediction model and used it to identify patients with acute SCI. Using Gene Ontology (GO) analysis and gene set enrichment analysis (GSEA), we discovered that immune-related biological processes, such as T cell receptor signaling pathway, were suppressed, whereas chemokine-related signaling pathways were activated after acute SCI. Immune infiltration analysis performed using single sample GSEA and CIBERSORT suggested that Tfh cell function was significantly correlated with the CCR7 expression levels and was considerably reduced after acute SCI. Acute SCI was divided into two subtypes, and we integrated multiple classifiers to analyze and elucidate the immunomodulatory relationships in both subtypes jointly. The results suggested that CCR7 suppresses the immunodeficiency phenotype by activating the chemokine signaling pathway in Tfh cells. In conclusion, CCR7 exhibits potential as a diagnostic marker for acute SCI.
The efficient generation of knockout microglia cells using a dual-sgRNA strategy by CRISPR/Cas9
Authors: Zhang Mengfei , Yi Fang , Wu Junjiao , Tang Yu
Published: October 2022

Gene deletion in microglia has become an important and exciting approach for studying neuroinflammation, especially after the development of the CRISPR/Cas9 system for genome editing during the last decade. In this study, we described a protocol for the highly efficient generation of knockout microglia cells using a dual-short guide RNA (sgRNA) strategy by CRISPR/Cas9. Leucine-rich repeat kinase 2 (LRRK2), a pathogenic gene of Parkinson's disease (PD), has played versatile roles during the disease development. Despite many key insights into LRRK2 studies, the normal and disease-related functions of LRRK2 in microglia and neuroinflammation remain to be fully investigated. Given the importance of LRRK2 in PD pathogenesis, we designed and applied the protocol to target LRRK2. Specifically, we designed two sgRNAs targeting the N terminus of LRRK2, spanning the 5' untranslated region (UTR) and exon 1, and screened knockout cells by single-cell expansion. In practice, the dual-sgRNA system can facilitate in obtaining knockout cells in a more convenient, rapid, and accurate way. Candidate knockout cells can be easily distinguished by genomic PCR and running on agarose gels, based on the different band sizes. Successful knockouts were further verified by Sanger sequencing and Western blot. Using this protocol, we obtained an LRRK2-deficient microglia cell line, which was characterized by longer cellular processes, enhanced adhesion, and weakened migration capacity. The knockout microglia may further serve as an important cellular tool to reveal conserved and novel aspects of LRRK2 functions in the development and progression of PD. Our protocol using dual-sgRNA targeting guarantees > 60% targeting efficiency and could also be applied to targeting other genes/loci, especially non-coding RNAs and regulatory elements.
Drunken lipid membranes, not drunken SNARE proteins, promote fusion in a model of neurotransmitter release
Authors: Coffman Robert E. , Kraichely Katelyn N. , Kreutzberger Alex J. B. , Kiessling Volker , Tamm Lukas K. , Woodbury Dixon J.
Published: October 2022

Alcohol affects many neuronal proteins that are upstream or down-stream of synaptic vesicle fusion and neurotransmitter release. Less well studied is alcohol’s effect on the fusion machinery including SNARE proteins and lipid membranes. Using a SNARE-driven fusion assay we show that fusion probability is significantly increased at 0.4% v/v (68 mM) ethanol; but not with methanol up to 10%. Ethanol appears to act directly on membrane lipids since experiments focused on protein properties [circular dichroism spectrometry, site-directed fluorescence interference contrast (sdFLIC) microscopy, and vesicle docking results] showed no significant changes up to 5% ethanol, but a protein-free fusion assay also showed increased lipid membrane fusion rates with 0.4% ethanol. These data show that the effects of high physiological doses of ethanol on SNARE-driven fusion are mediated through ethanol’s interaction with the lipid bilayer of membranes and not SNARE proteins, and that methanol affects lipid membranes and SNARE proteins only at high doses.
Folate vitamers in the Australian green plum: Through growth and ripening and across locations
Authors: Fyfe Selina , Hong Hung , Schirra Horst Joachim , Smyth Heather E. , Sultanbawa Yasmina , Rychlik Michael
Published: October 2022

The green plum is a native fruit of Australia that grows on the tree Buchanania obovata. This study aimed to confirm the high level of folate in green plums by analyzing a large number of ripe samples from multiple locations and to understand how folate vitamers change as the fruit grows through maturity stages. This study analyzed green plums for five vitamers of folate, H4folate, 5-CH3-H4folate, 5-CHO-H4folate, 10-CHO-PteGlu, and PteGlu (folic acid) using a stable isotope dilution assay on a liquid chromatograph mass spectrometer (LC-MS). Green plums were tested from four locations, two harvests and five maturity stages. Another 11 ripe samples, each from different tree clumps from one location, were also tested as were ripe red-colored green plums. The results show the 5-CH3-H4folate in green plum increases and accumulates in the fruit through development, ripening and senescence. The ripe green plums contain between 82.4 ± 5.5 and 149.4 ± 10.7 μg/100 g Fresh Weight (FW). The red-colored green plums are even higher in folate, with total folate measured as 192.5 ± 7.0 and 293.7 ± 27.4 μg/100 g FW, and further analysis of them is suggested. There is some variation in amounts of folate between fruit from different locations and sets of trees, but all ripe green plums tested are considered good dietary sources of folate.
A novel DNA methylation marker to identify lymph node metastasis of colorectal cancer
Authors: Yu Yingdian , Xue Wenyuan , Liu Zefeng , Chen Shang , Wang Jun , Peng Quanzhou , Xu Linhao , Liu Xin , Cui Chunhui , Fan Jian-Bing
Published: October 2022

Lymph node metastasis (LNM) of colorectal cancer (CRC) is an important factor for both prognosis and treatment. Given the deficiencies of conventional tests, we aim to discover novel DNA methylation markers to efficiently identify LNM status of CRC. In this study, genome-wide methylation sequencing was performed in a cohort (n=30) using fresh CRC tissue to discover differentially methylated markers. These markers were subsequently validated with fluorescence quantitative PCR in a cohort (n=221), and the optimal marker was compared to conventional diagnostic methods. Meanwhile, immunohistochemistry was used to verify the effectiveness of the antibody corresponding to this marker in a cohort (n=56). LBX2 achieved an AUC of 0.87, specificity of 87.3%, sensitivity of 75.7%, and accuracy of 81.9%, which outperformed conventional methods including imaging (CT, PET-CT) with an AUC of 0.52, CA199 with an AUC of 0.58, CEA with an AUC of 0.56. LBX2 was also superior to clinicopathological indicators including the depth of tumor invasion and lymphatic invasion with an AUC of 0.61and 0.63 respectively. Moreover, the AUC of LBX2 antibody was 0.84, which was also better than these conventional methods. In conclusion, A novel methylation marker LBX2 could be used as a simple, cost-effective, and reliable diagnostic method for LNM of CRC.
Atypical metanephric adenoma: Shares similar histopathological features and molecular changes of metanephric adenoma and epithelial-predominant Wilms’ tumor
Authors: Yin Xiaoxue , Zhang Xingming , Pan Xiuyi , Tan Junya , Zheng Linmao , Zhou Qiao , Chen Ni
Published: October 2022

Background

Metanephric adenomas (MAs) are rare, benign renal tumors. Wilms’ tumors (WTs) are malignant embryonic tumors that originated from nephrogenic blastemal cells. However, some tumors have similar morphology to both MA and epithelial-predominant WT, which makes differential diagnosis difficult. We aimed to analyze the morphological, immunophenotypic and molecular changes in overlapping cases to explore their attribution.

Methods and results

Twenty MAs, ten WTs, and nine cases with MA/WT overlapping histological features were studied. Twenty tumors demonstrated the typical morphological spectrum of MA with high cellularity and were composed of tightly packed small, uniform, round acini with a lower Ki67 index. Almost all MAs (94.7%, 18/19) were detected with BRAF V600E mutation. The ten WTs were epithelial-predominant WTs with glands, rosettes and glomerular structures, which also showed a higher Ki-67 index (up to 60%), invasive growth patterns, and a lack of BRAF mutation. However, the other nine overlapping cases showed two components: typical MA-like areas and epithelial WT-like areas. The cells of the WT-like areas were tubular, columnar and showed marked cytological atypia, with a Ki-67 proliferative index of up to 30%. The immunophenotype of these overlapping lesions was not significantly different from that of typical MA and they positively expressed WT1 and CD57. The BRAF V600E mutation was detected in both WT-like and MA-like areas in nine overlapping tumors. The follow-up data of 31 patients were analyzed, with a median follow-up time of 66 months (range, 8-45 months). Even though most patients with WT underwent radiotherapy or chemotherapy after surgery, two died, and one had liver metastasis. No MA or overlapping cases showed any evidence of recurrence or metastasis after surgery.

Conclusions

The molecular changes in tumors with overlapping morphological features were the same as those of typical MA; thus, we think that these tumors should be classified as MA and further called atypical MA. It is important to note that atypical MA is not a neglected subtype of MA. It possesses different histological morphology and a higher Ki-67 index but has the common imaging characteristics, immunophenotype and gene expression as typical MA, and patients usually have a good prognosis.
Patient-derived primary breast cancer cells and their potential for predicting sensitivity to chemotherapy
Authors: Mou Yajun , Huang Jianjun , Yang Wenxiu , Wan Yu , Pu Zhenhong , Zhang Junhong , Liu Jinting , Li Qing , Zhang Peipei , Tian Yuan , Yang Hui , Cui Yi , Hu Pingsheng , Dou Xiaowei
Published: October 2022

Chemotherapy resistance exposes patients to side effects and delays the effect of therapy in patients. So far, there are no predictive tools to predict resistance to chemotherapy and select sensitive chemotherapeutic drugs for the patient. Here, we aim to develop an in-vitro primary cell culture model from breast cancer patients to predict sensitivity to chemotherapy. We created the primary breast cancer cell medium BCMI and culture system with higher efficiency of the model establishment. Immunofluorescence staining of ERa, PR and HER2 were done to identify the primary breast cancer cell from the counterpart breast cancer patient. The killing assay showed that these primary breast cancer cells responded differently to doxorubicin and pirarubicin treatment. These results indicate that our established primary breast cancer cell model holds great promise for predicting breast cancer sensitivity to chemotherapy drugs.
Nomogram development and validation to predict Ki-67 expression of hepatocellular carcinoma derived from Gd-EOB-DTPA-enhanced MRI combined with T1 mapping
Authors: Liu Ziwei , Yang Shaomin , Chen Xinjie , Luo Chun , Feng Jieying , Chen Haixiong , Ouyang Fusheng , Zhang Rong , Li Xiaohong , Liu Wei , Guo Baoliang , Hu Qiugen
Published: October 2022

Objective

As an important biomarker to reflect tumor cell proliferation and tumor aggressiveness, Ki-67 is closely related to the high early recurrence rate and poor prognosis, and pretreatment evaluation of Ki-67 expression possibly provides a more accurate prognosis assessment and more better treatment plan. We aimed to develop a nomogram based on gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (Gd-EOB-DTPA)-enhanced magnetic resonance imaging (MRI) combined with T1 mapping to predict Ki-67 expression in hepatocellular carcinoma (HCC).

Methods

This two-center study retrospectively enrolled 148 consecutive patients who underwent preoperative Gd-EOB-DTPA-enhanced MRI T1 mapping and surgically confirmed HCC from July 2019 to December 2020. The correlation between quantitative parameters from T1 mapping, ADC, and Ki-67 was explored. Three cohorts were constructed: a training cohort (n = 73) and an internal validation cohort (n = 31) from Shunde Hospital of Southern Medical University, and an external validation cohort (n = 44) from the Sixth Affiliated Hospital, South China University of Technology. The clinical variables and MRI qualitative and quantitative parameters associational with Ki-67 expression were analyzed by univariate and multivariate logistic regression analyses. A nomogram was developed based on these associated with Ki-67 expression in the training cohort and validated in the internal and external validation cohorts.

Results

T1rt-Pre and T1rt-20min were strongly positively correlated with Ki-67 (r = 0.627, r = 0.607, P < 0.001); the apparent diffusion coefficient value was moderately negatively correlated with Ki-67 (r = -0.401, P < 0.001). Predictors of Ki-67 expression included in the nomogram were peritumoral enhancement, peritumoral hypointensity, T1rt-20min, and tumor margin, while arterial phase hyperenhancement (APHE) was not a significant predictor even included in the regression model. The nomograms achieved good concordance indices in predicting Ki-67 expression in the training and two validation cohorts (0.919, 0.925, 0.850), respectively.

Conclusions

T1rt-Pre and T1rt-20min had a strong positive correlation with the Ki-67 expression in HCC, and Gd-EOB-DTPA enhanced MRI combined with T1 mapping-based nomogram effectively predicts high Ki-67 expression in HCC.