bluuebunny/crossref_metadata_2025 huggingface dataset:
Combining functional metagenomics and glycoanalytics to identify enzymes that facilitate structural characterization of sulfated N-glycans
Authors: Chuzel Léa , Fossa Samantha L. , Boisvert Madison L. , Cajic Samanta , Hennig René , Ganatra Mehul B. , Reichl Udo , Rapp Erdmann , Taron Christopher H.
Published: August 2021

Background

Sulfate modification ofN-glycans is important for several biological functions such as clearance of pituitary hormones or immunoregulation. Yet, the prevalence of thisN-glycan modification and its functions remain largely unexplored. Characterization ofN-glycans bearing sulfate modifications is hampered in part by a lack of enzymes that enable site-specific detection ofN-glycan sulfation. In this study, we used functional metagenomic screening to identify enzymes that act upon sulfated N-acetylglucosamine (GlcNAc). Using multiplexed capillary gel electrophoresis with laser-induced fluorescence detection (xCGE-LIF) -based glycoanalysis we proved their ability to act upon GlcNAc-6-SO4onN-glycans.

Results

Our screen identified a sugar-specific sulfatase that specifically removes sulfate from GlcNAc-6-SO4when it is in a terminal position on anN-glycan. Additionally, in the absence of calcium, this sulfatase binds to the sulfated glycan but does not remove the sulfate group, suggesting it could be used for selective isolation of sulfatedN-glycans. Further, we describe isolation of a sulfate-dependent hexosaminidase that removes intact GlcNAc-6-SO4(but not asulfated GlcNAc) from a terminal position onN-glycans. Finally, the use of these enzymes to detect the presence of sulfatedN-glycans by xCGE-LIF is demonstrated.

Conclusion

The present study demonstrates the feasibility of using functional metagenomic screening combined with glycoanalytics to discover enzymes that act upon chemical modifications of glycans. The discovered enzymes represent new specificities that can help resolve the presence of GlcNAc-6-SO4inN-glycan structural analyses.
The m6A methyltransferase METTL3 controls epithelial-mesenchymal transition, migration and invasion of breast cancer through the MALAT1/miR-26b/HMGA2 axis
Authors: Zhao Chengpeng , Ling Xiaoling , Xia Yunxia , Yan Bingxue , Guan Quanlin
Published: August 2021

Background

Previous studies have revealed the key functions of N6-methyladenosine (m6A) modification in breast cancer (BC). MALAT1 as a highly m6A modified lncRNA associated with cancer development and metastasis, but the functional relevance of m6A methyltransferase and MALAT1 in BC is still unknown. Here, our study investigated the effects of the novel m6A methyltransferase METTL3 on epithelial-mesenchymal transition (EMT) in BC via the MALAT1/miR-26b/HMGA2 axis.

Methods

Firstly, we collected clinical BC samples and cultured BC cells, and detected mRNA and protein levels in the human samples and human cell lines by RT-qPCR and Western blot, respectively. Then, the binding of MALAT1 and miR-26b and the targeting relationship between miR-26b and HMGA2 were examined by dual-luciferase assay. Moreover, the binding of MALAT1 and miR-26b was tested by RNA pull down and RNA immunoprecipitation (RIP) assays. Methylated-RNA immunoprecipitation (Me-RIP) was used to detect the m6A modification level of MALAT1. The interaction of METTL3 and MALAT1 was detected by photoactivatable ribonucleoside-crosslinking immunoprecipitation (PAR-CLIP). Finally, effects on invasion and migration were detected by Transwell.

Results

In BC, the level of miR-26b was consistently low, while the levels of METTL3, MALAT1 and HMGA2 were high. Further experiments showed that METTL3 up-regulated MALAT1 expression by modulating the m6A modification of MALAT1, and that MALAT1 could promote the expression of HMGA2 by sponging miR-26b. In BC cells, we found that silencing METTL3 could inhibit EMT and tumor cell invasion by suppressing MALAT1. Furthermore, MALAT1 mediated miR-26b to target HMGA2 and promote EMT, migration, and invasion. In summary, METTL3 promoted tumorigenesis of BC via the MALAT1/miR-26b/HMGA2 axis.

Conclusions

Silencing METTL3 down-regulate MALAT1 and HMGA2 by sponging miR-26b, and finally inhibit EMT, migration and invasion in BC, providing a theoretical basis for clinical treatment of BC.
NCAPG promotes the progression of lung adenocarcinoma via the TGF-β signaling pathway
Authors: Wu Yun , Lin Ying , Pan Junfan , Tu Xunwei , Xu Yiquan , Li Hongru , Chen Yusheng
Published: August 2021

Background

Lung cancer has the highest case fatality rate among cancers because of uncontrolled proliferation and early metastasis of cancer cells in the lung tissue. This study aimed to clarify the role of the non-SMC condensin I complex, subunit G (NCAPG) in lung adenocarcinoma (LUAD), explore the mechanisms of its progression, and lay the foundation for the search for new biological markers.

Methods

We analyzed overlapping differentially expressed genes (DEGs) from three datasets; a protein–protein interaction (PPI) network was subsequently constructed and analyzed using Cytoscape. We then selected NCAPG for validation because of its poor prognosis and because it has not been sufficiently studied in the context of LUAD. Immunohistochemical analysis was used to detect the expression of NCAPG in LUAD tissues, and the relationships between NCAPG and clinical parameters were analyzed. In vitro and in vivo experiments were conducted to verify the role of NCAPG in LUAD. Finally, we studied the specific mechanism of action of NCAPG in LUAD.

Results

Through comprehensive analysis of the GSE43458, GSE75037, and The Cancer Genome Atlas databases, we identified 517 overlapping DEGs. Among them, NCAPG was identified as a hub gene. Immunohistochemical analysis revealed that NCAPG was strongly associated with the clinical stage, M-classification, and N-classification. Univariate and multivariate Cox regression analyses indicated that NCAPG was a prognostic risk factor for LUAD, while the in vitro experiments showed that NCAPG overexpression promoted proliferation, migration, invasion, and epithelial-mesenchymal transition. Furthermore, knockdown of NCAPG inhibited LUAD progression, both in vitro and in vivo. Mechanistically, NCAPG overexpression increased p-Smad2 and p-Smad3 expressions in the transforming growth factor β (TGF-β) signaling pathway. Additionally, rescue experiments indicated that TGF-β signaling pathway inhibitors could restore the effect of NCAPG overexpression in LUAD cells.

Conclusions

NCAPG may promote proliferation and migration via the TGF-β signaling pathway in LUAD.
NF-kB-dependent activation of STAT3 by H. pylori is suppressed by TFF1
Authors: Soutto Mohammed , Bhat Nadeem , Khalafi Shayan , Zhu Shoumin , Poveda Julio , Garcia-Buitrago Monica , Zaika Alexander , El-Rifai Wael
Published: August 2021

Background

H. pylori infection is the main risk factor for gastric cancer. In this study, we investigated H. pylori-mediated activation of STAT3 and NF-κB in gastric cancer, using in vitro and in vivo models.

Methods

To investigate the activation of NF-κB and STAT3 by H. pylori strains we used in vitro and in vivo mouse models, western blots, immunofluorescence, ChIP Assay, luciferase and quantitative real-time PCR assays.

Results

Following infection with H. pylori in vitro, we found an earlier phosphorylation of NF-kB-p65 (S536), followed by STAT3 (Y705). Immunofluorescence, using in vitro and in vivo models, demonstrated nuclear localization of NF-kB and STAT3, following H. pylori infection. NF-kB and STAT3 luciferase reporter assays confirmed earlier activation of NF-kB followed by STAT3. In vitro and in vivo models demonstrated induction of mRNA expression of IL-6 (p < 0.001), VEGF-α (p < 0.05), IL-17 (p < 0.001), and IL-23 (p < 0.001). Using ChIP, we confirmed co-binding of both NF-kB-p65 and STAT3 on the IL6 promoter. The reconstitution of Trefoil Factor 1 (TFF1) suppressed activation of NF-kB with reduction in IL6 levels and STAT3 activity, in response to H. pylori infection. Using pharmacologic (BAY11-7082) and genetic (IκB super repressor (IκBSR)) inhibitors of NF-kB-p65, we confirmed the requirement of NF-kB-p65 for activation of STAT3, as measured by phosphorylation, transcription activity, and nuclear localization of STAT3 in in vitro and in vivo models.

Conclusion

Our findings suggest the presence of an early autocrine NF-kB-dependent activation of STAT3 in response to H. pylori infection. TFF1 acts as an anti-inflammatory guard against H. pylori-mediated activation of pro-inflammatory networks.
NEK7 promotes gastric cancer progression as a cell proliferation regulator
Authors: Li Yi-Ke , Zhu Xiao-Ran , Zhan Yue , Yuan Wen-Zhen , Jin Wei-Lin
Published: August 2021

Background

Gastric cancer is one of the most common malignant tumors of the digestive system. However, its targeted therapy develops at a slow pace. Thus, exploring the mechanisms of the malignant behavior of gastric cancer cells is crucial to exploit its treatment. Mammalian never-in-mitosis A (NIMA)-related kinases (NEKs) are considered to play a significant role in cancer cell proliferation. However, no study has reported on NIMA family proteins in gastric cancer.

Methods

Bioinformatics analysis was employed to clarify the expression patterns of NEK1–NEK11 and their effects on prognosis. The effects of NEK7 on immune infiltration and NEK7 related pathways were also analyzed. At the cell level, 5-ethynyl-2-deoxyuridine, cell cycle, and Cell Counting Kit-8 assays were utilized to clarify the effect of NEK7 on gastric cancer cell proliferation. A mouse subcutaneous model revealed the regulating effect of NEK7 on gastric cancer cell proliferation in vivo.

Results

Bioinformatics analysis revealed that NEK7 is upregulated in gastric cancer and is related to poor prognosis. NEK7 is also related to T-stage, which is closely associated with cell proliferation. Further analysis showed that NEK7 was correlated with infiltration of multiple immune cells as well as gastric cancer-related pathways. Cell experiments indicated the promoting effect of NEK7 on cell proliferation, while the absence of NEK7 could lead to inhibition of gastric cancer proliferation and G1/S arrest.

Conclusion

NEK7 exerts a regulatory effect on cell proliferation and is closely related to tumor immune infiltration.
Molecular mechanisms of the microRNA-132 during tumor progressions
Authors: Moghbeli Meysam , Zangouei Amir Sadra , Nasrpour Navaii Zahra , Taghehchian Negin
Published: August 2021

Cancer as one of the leading causes of human deaths has always been one of the main health challenges in the world. Despite recent advances in therapeutic and diagnostic methods, there is still a high mortality rate among cancer patients. Late diagnosis is one of the main reasons for the high ratio of cancer related deaths. Therefore, it is required to introduce novel early detection methods. Various molecular mechanisms are associated with the tumor progression and metastasis. MicroRNAs (miRNAs) are a class of non-coding RNAs (ncRNAs) family that has important functions in regulation of the cellular processes such as cell proliferation, apoptosis, and tumor progression. Moreover, they have higher stability in body fluids compared with mRNAs which can be introduced as non-invasive diagnostic markers in cancer patients. MiR-132 has important functions as tumor suppressor or oncogene in different cancers. In the present review, we have summarized all of the studies which have been reported the role of miR-132 during tumor progressions. We categorized the miR-132 target genes based on their cell and molecular functions. Although, it has been reported that the miR-132 mainly functions as a tumor suppressor, it has also oncogenic functions especially in pancreatic tumors. MiR-132 mainly exerts its roles during tumor progressions by regulation of the transcription factors and signaling pathways. Present review clarifies the tumor specific molecular mechanisms of miR-132 to introduce that as an efficient non-invasive diagnostic marker in various cancers.
Assessment of turbulent blood flow and wall shear stress in aortic coarctation using image-based simulations
Authors: Perinajová Romana , Juffermans Joe F. , Mercado Jonhatan Lorenzo , Aben Jean-Paul , Ledoux Leon , Westenberg Jos J. M. , Lamb Hildo J. , Kenjereš Saša
Published: August 2021

In this study, we analyzed turbulent flows through a phantom (a 180$$^{\circ }$$∘bend with narrowing) at peak systole and a patient-specific coarctation of the aorta (CoA), with a pulsating flow, using magnetic resonance imaging (MRI) and computational fluid dynamics (CFD). For MRI, a 4D-flow MRI is performed using a 3T scanner. For CFD, the standard$$k-\epsilon $$k-ϵ, shear stress transport$$k-\omega $$k-ω, and Reynolds stress (RSM) models are applied. A good agreement between measured and simulated velocity is obtained for the phantom, especially for CFD with RSM. The wall shear stress (WSS) shows significant differences between CFD and MRI in absolute values, due to the limited near-wall resolution of MRI. However, normalized WSS shows qualitatively very similar distributions of the local values between MRI and CFD. Finally, a direct comparison between in vivo 4D-flow MRI and CFD with the RSM turbulence model is performed in the CoA. MRI can properly identify regions with locally elevated or suppressed WSS. If the exact values of the WSS are necessary, CFD is the preferred method. For future applications, we recommend the use of the combined MRI/CFD method for analysis and evaluation of the local flow patterns and WSS in the aorta.
‘Doing’ or ‘using’ intersectionality? Opportunities and challenges in incorporating intersectionality into knowledge translation theory and practice
Authors: Kelly Christine , Kasperavicius Danielle , Duncan Diane , Etherington Cole , Giangregorio Lora , Presseau Justin , Sibley Kathryn M. , Straus Sharon
Published: August 2021

Intersectionality is a widely adopted theoretical orientation in the field of women and gender studies. Intersectionality comes from the work of black feminist scholars and activists. Intersectionality argues identities such as gender, race, sexuality, and other markers of difference intersect and reflect large social structures of oppression and privilege, such as sexism, racism, and heteronormativity. The reach of intersectionality now extends to the fields of public health and knowledge translation. Knowledge translation (KT) is a field of study and practice that aims to synthesize and evaluate research into an evidence base and move that evidence into health care practice. There have been increasing calls to bring gender and other social issues into the field of KT. Yet, as scholars outline, there are few guidelines for incorporating the principles of intersectionality into empirical research. An interdisciplinary, team-based, national health research project in Canada aimed to bring an intersectional lens to the field of knowledge translation. This paper reports on key moments and resulting tensions we experienced through the project, which reflect debates in intersectionality: discomfort with social justice, disciplinary divides, and tokenism. We consider how our project advances intersectionality practice and suggests recommendations for using intersectionality in health research contexts. We argue that while we encountered many challenges, our process and the resulting co-created tools can serve as a valuable starting point and example of how intersectionality can transform fields and practices.
Impact of imprecise household location on effective coverage estimates generated through linking household and health provider data by geographic proximity: a simulation study
Authors: Carter Emily D. , Munos Melinda K.
Published: August 2021

Background

Geographic proximity is often used to link household and health provider data to estimate effective coverage of health interventions. Existing household surveys often provide displaced data on the central point within household clusters rather than household location. This may introduce error into analyses based on the distance between households and providers.

Methods

We assessed the effect of imprecise household location on quality-adjusted effective coverage of child curative services estimated by linking sick children to providers based on geographic proximity. We used data on care-seeking for child illness and health provider quality in Southern Province, Zambia. The dataset included the location of respondent households, a census of providers, and data on the exact outlets utilized by sick children included in the study. We displaced the central point of each household cluster point five times. We calculated quality-adjusted coverage by assigning each sick child to a provider’s care based on three measures of geographic proximity (Euclidean distance, travel time, and geographic radius) from the household location, cluster point, and displaced cluster locations. We compared the estimates of quality-adjusted coverage to each other and estimates using each sick child’s true source of care. We performed sensitivity analyses with simulated preferential care-seeking from higher-quality providers and randomly generated provider quality scores.

Results

Fewer children were linked to their true source of care using cluster locations than household locations. Effective coverage estimates produced using undisplaced or displaced cluster points did not vary significantly from estimates produced using household location data or each sick child’s true source of care. However, the sensitivity analyses simulating greater variability in provider quality showed bias in effective coverage estimates produced with the geographic radius and travel time method using imprecise location data in some scenarios.

Conclusions

Use of undisplaced or displaced cluster location reduced the proportion of children that linked to their true source of care. In settings with minimal variability in quality within provider categories, the impact on effective coverage estimates is limited. However, use of imprecise household location and choice of geographic linking method can bias estimates in areas with high variability in provider quality or preferential care-seeking.
Serum levels of Asprosin in patients diagnosed with coronary artery disease (CAD): a case-control study
Authors: Moradi Nariman , Fouani Fatima Zahraa , Vatannejad Akram , Bakhti Arani Abbas , Shahrzad Soraya , Fadaei Reza
Published: August 2021

Background

Coronary artery disease (CAD) is considered as a multi-faceted chronic inflammatory disease involving reduced blood supply to the myocardium as a result of accumulating lipids in the atrial walls. Visceral adiposity with disrupted release of adipokines play a key role in its pathogenesis. Asprosin is a newly identified fasting-induced glucogenic adipokine that has been related with metabolic disorders such as type II diabetes mellitus and polycystic ovary syndrome. The preset study sought to assess circulating asprosin in context of CAD.

Methods

In this study, serum levels of asprosin were determined in 88 CAD patients and 88 non-CAD healthy controls. Serum IL-6, TNF-α, asprosin and adiponectin were assessed using ELISA kits.

Results: Serum asprosin was found to be higher in CAD patients when compared to non-CAD subjects (7.84 ± 2.08 versus 5.02 ± 1.29 μg/mL, p <  0.001). Similarly, serum TNF-α, and IL-6 elevated in CAD group significantly (p <  0.001). However, circulating adiponectin diminished in CAD group when compared with non-CAD subjects (p < 0.001). Moreover, serum asprosin levels directly correlated with BMI, FBG, HOMA-IR, TG and TC. Logistic regression analyses showed that asprosin levels were associated with increased risk of developing CAD (odds ratio: 3.01, 95% CI: 2.16, 4.20 and p < 0.001), after adjusting for potential confounders (age, sex and BMI).

Conclusions

The present study findings suggested a possible relation of serum asprosin with the pathogenesis of CAD, in particular through insulin resistance and dyslipidemia.
Food access among people who inject drugs in West Virginia
Authors: Rouhani Saba , Allen Sean T. , Whaley Sara , White Rebecca Hamilton , O’Rourke Allison , Schneider Kristin E. , Kilkenny Michael E. , Weir Brian W. , Sherman Susan G.
Published: August 2021

Background

The substance use epidemic in the United States continues to drive high levels of morbidity and mortality, particularly among people who inject drugs (PWID). Poor access to food often co-occurs with drug use and contributes to associated sequelae, such as risks for HIV and diabetes. The objective of this study was to examine factors associated with adequate food access among PWID in a rural Appalachian community.

Methods

Cross-sectional surveys were used to collect data among PWID aged 18 and older in Cabell County, West Virginia. Frequency of hunger and sociodemographic, structural and drug use characteristics were measured. Adequate food access was defined as reporting ‘never’ going to bed hungry at night in the past six months. Pearson’s χ2 and t-tests and multivariable logistic regression were used to identify factors associated with food access.

Results

Only 71 individuals (17%) reported never going to bed hungry at night in the past six months. Adjusted odds of having adequate food access were higher among PWID who completed high school (aOR 2.94; P = 0.010) and usually used drugs alone (aOR 1.97; P = 0.025), and lower among PWID who were female (aOR 0.51; P = 0.037), experienced homelessness (aOR 0.23, P < 0.001), were recently arrested (aOR 0.50 P = 0.047), and engaged in receptive sharing of injection equipment (aOR 0.52, P = 0.035).

Conclusions

We found extremely low food access in a population of PWID in Appalachia who are vulnerable to overdose and infectious disease transmission. Integrated interventions promoting food access are needed to improve the public health and wellbeing of people who inject drugs in Appalachia.
Content validation of the Well-being of Older People measure (WOOP)
Authors: Hackert Mariska Q. N. , van Exel Job , Brouwer Werner B. F.
Published: August 2021

Background

Valid measures of the well-being of older people are important for the evaluation of health and social care services. The nine-item Well-being of Older People measure (WOOP) was based on a novel framework derived from a recent Q-methodology study, and was developed to capture a comprehensive set of well-being domains relevant to older people, as identified by themselves. This study introduces the WOOP and describes the qualitative assessment of its feasibility and content validity.

Methods

Between December 2017 and January 2018, a sampling agency retrieved data from 269 adults aged 65 years and older in the Netherlands. Using an online survey, participants were asked to complete the WOOP and to indicate the importance of each item to their well-being. Open-ended questions were used to collect information about participants’ own definition of well-being, their interpretation of the items of the WOOP, and their assessment of the descriptions and response options provided with each item. Data were analysed using inductive content analysis with the software package ATLAS.ti.

Results

The WOOP closely resembled respondents’ own description of what well-being means to them. The majority of the respondents reported no important well-being aspects to be missing from the WOOP, and indicated all WOOP items to be at least ‘reasonably important’ to their well-being. Many linked the WOOP items to well-being aspects as intended, and only a few had suggestions for improving the items’ descriptions and response options.

Conclusions

Given these results, all nine items were retained, and no items were added to the measure. Based on respondents’ feedback, minor changes were made to the wording of some descriptions and response options of items. Concluding, the feasibility and content validity of the WOOP seem satisfactory. Further validation of this new measure is required, in different health and social care settings and among subgroups of older people with potentially different views on what constitutes well-being.
Association between cardiovascular risk-factors and venous thromboembolism in a large longitudinal study of French women
Authors: MacDonald C. J. , Madika A. L. , Lajous M. , Canonico M. , Fournier A. , Boutron-Ruault M. C.
Published: August 2021

Background

Previous studies have shown conflicting results regarding the influence of cardiovascular risk-factors on venous thromboembolism. This study aimed to determine if these risk-factors, i.e. physical activity, smoking, hypertension, dyslipidaemia, and diabetes, were associated with the risk of venous thromboembolism, and to determine if these associations were confounded by BMI.

Methods

We used data from the E3N cohort study, a French prospective population-based study initiated in 1990, consisting of 98,995 women born between 1925 and 1950. From the women in the study we included those who did not have prevalent arterial disease or venous thromboembolism at baseline; thus 91,707 women were included in the study. Venous thromboembolism cases were self-reported during follow-up, and verified via specific mailings to medical practitioners or via drug reimbursements for anti-thrombotic medications. Hypertension, diabetes and dyslipidaemia were self-reported validated against drug reimbursements or specific questionnaires. Physical activity, and smoking were based on self-reports. Cox-models, adjusted for BMI and other potential risk-factors were used to determine hazard ratios for incident venous thromboembolism.

Results

During 1,897,960 person-years (PY), 1, 649 first incident episodes of thrombosis were identified at an incidence rate of 0.9 per 1000 PY. This included 505 cases of pulmonary embolism and 1144 cases of deep vein thrombosis with no evidence of pulmonary embolism. Hypertension, dyslipidaemia, diabetes, smoking and physical activity were not associated with the overall risk of thrombosis after adjustment for BMI.

Conclusions

Traditional cardiovascular risk factors were not associated with the risk of venous thromboembolism after adjustment for BMI. Hypertension, dyslipidaemia and diabetes may not be risk-factors for venous thromboembolism.
Integrative analysis and experiments to explore angiogenesis regulators correlated with poor prognosis, immune infiltration and cancer progression in lung adenocarcinoma
Authors: Cai Songhua , Guo Xiaotong , Huang Chujian , Deng Youjun , Du Longde , Liu Wenyi , Yang Chenglin , Zhao Hongbo , Ma Kai , Wang Lixu , He Jie , Yu Zhentao
Published: August 2021

Angiogenesis is the process of capillary sprouting from pre-existing vessels and it plays a critical role in the carcinogenic process of lung adenocarcinoma (LUAD). However, the association of angiogenesis regulators with the prognosis and progression of LUAD needs to be further elucidated. In this study, we adopted differential expression analysis, Cox proportional hazards (PH) regression analysis and experimental validation to identify angiogenesis regulators correlated with a poor prognosis, immune infiltration and cancer progression in LUAD. These results showed that the diagnostic and prognostic models based on COL5A2 and EPHB2 served as independent biomarkers with superior predictive ability. The patients in the high-risk group exhibited a worse prognosis in the TCGA cohort (P < 0.001, HR = 1.72, 95% CI 1.28–2.30), GSE310210 cohort (P = 0.005, HR = 2.87, 95% CI 1.46–5.61), and GSE31019 cohort (P = 0.01, HR = 2.14, 95% CI 1.19–3.86) than patients in the low-risk group. The high prognostic risk patients had a higher TMB (P < 0.001); higher fractions of M0 macrophages, neutrophils, NK cells resting, and T cells CD4 memory activated (P < 0.05); and higher expression of immune checkpoints PD-1, PDL-1, PDL-2, and B7H3 (P < 0.001). Patients in the high-risk group were more sensitive to chemotherapeutic drugs and molecular targeted drugs such as cisplatin, doxorubicin, gefitinib, and bosutinib (P < 0.0001). In addition, inhibition of COL5A2 and EPHB2 effectively suppressed the proliferation and migration of LUAD cells. The current study identified angiogenesis regulators as potential biomarkers and therapeutic targets for LUAD and may help to further optimize cancer therapy.
Integrated analysis of competing endogenous RNA networks in peripheral blood mononuclear cells of systemic lupus erythematosus
Authors: Song Wencong , Qiu Jie , Yin Lianghong , Hong Xiaoping , Dai Weier , Tang Donge , Liu Dongzhou , Dai Yong
Published: August 2021

Background

Systemic lupus erythematosus (SLE) is an autoimmune disease with a complicated pathogenesis, and its aetiology has not been clearly unveiled. The lack of effective diagnosis and treatment methods makes it necessary to explore the molecular mechanism of SLE. We aimed to identify some critical signalling pathways and key competing endogenous RNAs (ceRNAs) underlying the molecular mechanism of SLE and to map out the systematic signalling networks by integrating the data on different kinds of RNAs.

Methods

Peripheral blood mononuclear cells (PBMCs) were collected from both SLE patients and healthy subjects, RNA was extracted from the PBMCs, and RNA libraries including ribosomal RNA-depleted strand-specific libraries and small RNA libraries were built for deep RNA sequencing (RNA-seq). RNA-seq yielded differential expression profiles of lncRNAs/circRNAs/miRNAs/mRNAs related to SLE. The DAVID database (v. 6.8) was employed for Gene Ontology (GO) and KEGG pathway analysis. ceRNA networks (circRNA/lncRNA-miRNA-mRNA) were constructed and visualized using Cytoscape software (v. 3.5.0). The TargetScan and miRanda databases were used to predict target relationships in ceRNA networks. qRT-PCR was used to verify our data.

Results

Differential expression of ceRNAs related to SLE was detected in SLE patients’ PBMCs: 644 mRNAs (384 upregulated, 260 downregulated), 326 miRNAs (223 upregulated, 103 downregulated), 221 lncRNAs (79 upregulated, 142 downregulated), and 31 circRNAs (21 upregulated, 10 downregulated). We drew ceRNA signalling networks made up of the differentially expressed mRNAs/miRNAs/lncRNAs/circRNAs mentioned above, and the hub genes included IRF5, IFNAR2, TLR7, IRAK4, STAT1, STAT2, C2, and Tyk2. These hub genes were involved in ceRNA signalling pathways, such as the IL-17 signalling pathway and type I interferon signalling pathway.

Conclusions

We explored the differential expression profiles of various kinds of ceRNAs and integrated signalling networks constructed by ceRNAs. Our findings offer new insights into the pathogenesis of SLE and hint at therapeutic strategies.
Clinical remission and subsequent relapse in patients with juvenile idiopathic arthritis: predictive factors according to therapeutic approach
Authors: Castillo-Vilella Mireia , Giménez Nuria , Tandaipan Jose Luis , Quintana Salvador , Modesto Consuelo
Published: August 2021

Background

Juvenile idiopathic arthritis constitutes a significant cause of disability and quality of life impairment in pediatric and adult patients. The aim of this study was to ascertain clinical remission (CR) and subsequent relapse in juvenile idiopathic arthritis (JIA) patients, according to therapeutic approach and JIA subtype. Evidence in literature regarding its predictors is scarce.

Methods

We conducted an observational, ambispective study. Patients diagnosed of JIA, treated with synthetic and/or biologic disease modifying antirheumatic drugs (DMARD) were included and followed-up to December 31st, 2015. Primary outcome was clinical remission defined by Wallace criteria, both on and off medication. In order to ascertain CR according to therapeutic approach, DMARD treatments were divided in four groups: 1) synthetic DMARD (sDMARD) alone, 2) sDMARD combined with another sDMARD, 3) sDMARD combined with biologic DMARD (bDMARD), and 4) bDMARD alone.

Results

A total of 206 patients who received DMARD treatment were included. At the time the follow-up was completed, 70% of the patients in the cohort had attained CR at least once (144 out of 206), and 29% were in clinical remission off medication (59 out of 206).

According to treatment group, CR was more frequently observed in patients treated with synthetic DMARD alone (53%). Within this group, CR was associated with female sex, oligoarticular persistent subtypes, ANA positivity, Methotrexate treatment and absence of HLA B27, comorbidities and DMARD toxicity. 124 DMARD treatments (62%) were withdrawn, 64% of which relapsed. Lower relapse rates were observed in those patients with persistent oligoarticular JIA (93%) when DMARD dose was tapered before withdrawal (77%).

Conclusions

More than two thirds of JIA patients attained CR along the 9 years of follow-up, and nearly one third achieved CR off medication. Females with early JIA onset, lower active joint count and ANA positivity were the ones achieving and sustaining remission more frequently, especially when receiving synthetic DMARD alone and in the absence of HLA B27, comorbidities or previous DMARD toxicity.
Anti-AQP4 autoantibodies promote ATP release from astrocytes and induce mechanical pain in rats
Authors: Ishikura Teruyuki , Kinoshita Makoto , Shimizu Mikito , Yasumizu Yoshiaki , Motooka Daisuke , Okuzaki Daisuke , Yamashita Kazuya , Murata Hisashi , Beppu Shohei , Koda Toru , Tada Satoru , Shiraishi Naoyuki , Sugiyama Yasuko , Miyamoto Katsuichi , Kusunoki Susumu , Sugimoto Tomoyuki , Kumanogoh Atsushi , Okuno Tatsusada , Mochizuki Hideki
Published: August 2021

Background

Intractable neuropathic pain is a common symptom of neuromyelitis optica spectrum disorder (NMOSD). However, the underlying mechanism of NMOSD pain remains to be elucidated. In this study, we focused on ATP, which is one of the damage-associated molecular patterns, and also a well-recognized molecule involved in peripheral neuropathic pain.

Methods

We assessed the development of pain symptoms by injecting anti-AQP4 recombinant autoantibodies (rAQP4 IgG) into rat spinal cords. We incubated HEK293 cells expressing AQP4 (HEK-AQP4) and rat astrocytes with rAQP4 IgG and assessed the level of ATP in the supernatant. We performed transcriptome analysis of the spinal cords injected with rAQP4 IgG. Pharmacological inhibition was also applied to investigate the involvement of ATP in the development of neuropathic pain in our rat model. The ATP concentration within the cerebrospinal fluid was examined in patients with NMOSD and other neurological diseases.

Results

Development of mechanical allodynia was confirmed in rAQP4 IgG–treated rats. AQP4-Ab–mediated extracellular ATP release from astrocytes was observed in vitro, and pharmacological inhibition of ATP receptor reversed mechanical allodynia in the rAQP4 IgG–treated rats. Furthermore, transcriptome analysis revealed elevation of gene expressions related to several ATP receptors including P2rx4 and IL1B in the spinal cord of rAQP4 IgG–treated rats. In patients, CSF ATP concentration was significantly higher in the acute and remission phase of NMOSD than in multiple sclerosis or other neurological disorders.

Conclusion

Anti-AQP4 antibody was shown to induce the release of extracellular ATP from astrocytes. The ATP-mediated development of mechanical allodynia was also suggested in rats treated with anti-AQP4 antibody. Our study indicates the pivotal role of ATP in the pain mechanism of NMOSD.
Repeated neonatal sevoflurane induced neurocognitive impairment through NF-κB-mediated pyroptosis
Authors: Dai Jing , Li Xue , Wang Cai , Gu Shuxin , Dai Lei , Zhang Jingyun , Fan Yunxia , Wu Jing
Published: August 2021

Background

Exposure to general anesthesia (GA) during the postnatal period is associated with neuroinflammation and long-term neurocognitive impairment in preclinical and clinical settings. Pyroptosis is a novel type of programmed cell death that, along with inflammation, has been found to play an important role in the mechanism of diverse neurological diseases. However, its roles in GA-induced neuroinflammation and neurocognitive impairment in the developing brain have not been investigated.

Methods

Rats at postnatal day 6 or primary hippocampal neurons at 9 days in vitro received 3% sevoflurane for 2 h daily for three consecutive days. A pharmacological inhibitor of nuclear factor (NF)-κB (BAY 11-7082) was administered to suppress NF-κB activation. Histological and biochemical analyses were performed to assess the pyroptosis as well as neuronal and synaptic damage both in vivo and in vitro. In addition, behavioral tests were performed to evaluate neurocognitive ability in rats.

Results

Repeated sevoflurane exposure activated NF-κB-mediated pyroptosis and neuroinflammation in the hippocampus in developing rats, damaged the neuronal morphology and synaptic integrity, and induced neurocognitive impairment in rats. BAY 11-7082 treatment suppressed the activation of pyroptosis, attenuated the neuronal and synaptic damage, and ameliorated the neurocognitive impairment induced by repeated sevoflurane administration to developing rats.

Conclusions

Repeated sevoflurane GA may induce neuroinflammation and neurocognitive impairment in developing rats via the activation of NF-κB-mediated pyroptosis. Our findings characterize a novel role of pyroptosis as a potential therapeutic target in neuroinflammation after repeated neonatal GA.
On the relationship between inhibition and receptor occupancy by nondepolarizing neuromuscular blocking drugs
Authors: Hoshino Hikaru , Furutani Eiko
Published: August 2021

Background

Nondepolarizing neuromuscular blocking drugs (NDNBs) are clinically used to produce muscle relaxation during general anesthesia. To better understand clinical properties of NDNBs, comparative in vitro pharmacologic studies have been performed. In these studies, a receptor binding model, which relies on the assumption that the inhibition, i.e., the effect of an NDNB, is proportional to the receptor occupancy by the drug, has been effectively used to describe obtained experimental data. However, it has not been studied in literature under which conditions the above assumption can be justified nor the assumption still holds in vivo. The purpose of this study is to explore the in vivo relationship between the inhibition and the receptor occupancy by an NDNB and to draw implications on how in vitro experimental results can be used to discuss the in vivo properties of NDNBs.

Methods

An ordinary differential equation model is employed to simulate physiologic processes of the activation of receptors by acetylcholine (ACh) as well as inhibition by an NDNB. With this model, the degree of inhibition is quantified by the fractional amount of receptors that are not activated by ACh due to the presence of an NDNB. The results are visualized by plotting the fractional amounts of the activated receptors as a function of the receptor occupancy.

Results

Numerical investigations reflecting in vivo conditions show that the degree of inhibition is not proportional to the receptor occupancy, i.e., there is a nonlinear relationship between the inhibition and the receptor occupancy. However, under a setting of high concentration of ACh reflecting a typical situation of in vitro experiments, the relationship between the inhibition and the receptor occupancy becomes linear, suggesting the validity of the receptor binding model. Also, it is found that the extent of nonlinearity depends on the selectivity of NDNBs for the two binding sites of the receptors.

Conclusions

While the receptor binding model may be effective for estimating affinity of an NDNB through in vitro experiments, these models do not directly describe in vivo properties of NDNBs, because the nonlinearity between the inhibition and the receptor occupancy causes the modulation of the resultant concentration-effect relationships of NDNBs.
Modern contraceptive use among women in need of family planning in India: an analysis of the inequalities related to the mix of methods used
Authors: Ewerling Fernanda , McDougal Lotus , Raj Anita , Ferreira Leonardo Z. , Blumenberg Cauane , Parmar Divya , Barros Aluisio J. D.
Published: August 2021

Objective

To evaluate the type of contraceptives used by women in need of family planning in India and the inequalities associated with that use according to women's age, education, wealth, subnational region of residence and empowerment level.

Methods

Using data from the Indian National Family and Health Survey-4 (2015–2016), we evaluated the proportion of partnered women aged 15–49 years with demand for family planning satisfied (DFPS) with modern contraceptive methods. We also explored the share of each type of contraception [short- (e.g., condom, pill) and long-acting (i.e., IUD) reversible contraceptives and permanent methods] and related inequalities.

Results

The majority (71.8%; 95% CI 71.4–72.2) of women in need of contraception were using a modern method, most (76.1%) in the form of female sterilization. Condom and contraceptive pill were the second and third most frequently used methods (11.8% and 8.5%, respectively); only 3.2% reported IUD. There was a nearly linear exchange from short-acting to permanent contraceptive methods as women aged. Women in the poorest wealth quintile had DFPS with modern methods at least 10 percentage points lower than other women. We observed wide geographic variation in DFPS with modern contraceptives, ranging from 23.6% (95% CI 22.1–25.2) in Manipur to 93.6% (95% CI 92.8–94.3) in Andhra Pradesh. Women with more accepting attitudes towards domestic violence and lower levels of social independence had higher DFPS with modern methods but also had higher reliance on permanent methods. Among sterilized women, 43.2% (95% CI 42.7–43.7) were sterilized before age 25, 61.5% (95% CI 61.0–62.1) received monetary compensation for sterilization, and 20.8% (95% CI 20.3–21.3) were not informed that sterilization prevented future pregnancies.

Conclusion

Indian family planning policy should prioritize women-centered care, making reversible contraceptive methods widely available and promoted.
Tenofovir alafenamide nephrotoxicity: a case report and literature review
Authors: Ueaphongsukkit Thornthun , Gatechompol Sivaporn , Avihingsanon Anchalee , Surintrspanont Jerasit , Iampenkhae Kroonpong , Avihingsanon Yingyos , Udomkarnjananun Suwasin
Published: August 2021

Background

Tenofovir alafenamide (TAF), a novel prodrug of tenofovir (TFV), has become the preferred drug for the treatment of HIV-1 and chronic hepatitis B infection in clinical practice. Results from clinical trials showed that it had better renal and bone mineral outcomes compared to tenofovir disoproxil fumarate (TDF). However, as we have seen with TDF, side effects from the new medication can be more prevalent and recognized after extensive use in real world situations. Sporadic cases of acute kidney injury in patients using TAF have started to emerge.

Case presentation

We report a case of 49-year-old Thai, HIV treatment-experienced female with hypertension presented with worsening renal function after switching her antiretroviral regimen from TDF, emtricitabine (FTC), and lopinavir/ritonavir (LPV/r) to TAF, FTC and dolutegravir (DTG) for 3 months. Kidney biopsy showed distinctive picture of tenofovir nephrotoxicity with acute tubular injury and mitochondrial injury. The possible causes of acute kidney injury and nephrotoxicity from TAF for this patient were discussed. We have extensively reviewed all published case reports of TAF-associated nephrotoxicity and summarized the essential information in this article.

Conclusion

Although TAF has less nephrotoxicity compared with TDF; renal function should always be monitored after the initiation of both drugs. Future large cohort studies are required to identify the risk factors of TAF-associated nephrotoxicity and to design an effective preventive strategy.
Cytotoxicity analysis of biomass combustion particles in human pulmonary alveolar epithelial cells on an air–liquid interface/dynamic culture platform
Authors: Ke Shaorui , Liu Qi , Zhang Xinlian , Yao Yuhan , Yang Xudong , Sui Guodong
Published: August 2021

Background

Exposure to indoor air pollution from solid fuel combustion is associated with lung diseases and cancer. This study investigated the cytotoxicity and molecular mechanisms of biomass combustion-derived particles in human pulmonary alveolar epithelial cells (HPAEpiC) using a platform that combines air–liquid interface (ALI) and dynamic culture (DC) systems.

Methods

HPAEpiC were cultured on the surface of polycarbonate (PC) membranes on the ALI–DC platform. The cells were sprayed with an aerosolized solution of biomass combustion soluble constituents (BCSCs) and simultaneously nourished with culture medium flowing beneath the permeable PC membranes. The ALI–DC method was compared with the traditional submerged culture approach. BCSC particle morphology and dosages deposited on the chip were determined for particle characterization. Flow cytometry, scanning electron microscopy, and transmission electron microscopy were used to investigate the apoptosis rate of HPAEpiC and changes in the cell ultrastructure induced by BCSCs. Additionally, the underlying apoptotic pathway was examined by determining the protein expression levels by western blotting.

Results

Scanning electron microscope images demonstrated that the sample processing and delivering approach of the ALI–DC platform were suitable for pollutant exposure. Compared with the submerged culture method, a significant decline in cell viability and increase in apoptosis rate was observed after BCSC exposure on the ALI–DC platform, indicating that the ALI–DC platform is a more sensitive system for investigating cytotoxicity of indoor air pollutants in lung cells. The morphology and ultrastructure of the cells were damaged after exposure to BCSCs, and the p53 pathway was activated. The Bcl-2/Bax ratio was reduced, upregulating caspase-9 and caspase-3 expression and subsequently inducing apoptosis of HPAEpiC. The addition ofN-acetyl cysteine antioxidant significantly alleviated the cytotoxicity induced by BCSCs.

Conclusion

A novel ALI–DC platform was developed to study the cytotoxicity of air pollutants on lung cells. Using the platform, we demonstrated that BCSCs could damage the mitochondria, produce reactive oxygen species, and activate p53 in HPAEpiC, ultimately inducing apoptosis.
Perceived stress and post-traumatic stress disorder symptoms among intensive care unit staff caring for severely ill coronavirus disease 2019 patients during the pandemic: a national study
Authors: Kader Nisha , Elhusein Bushra , Chandrappa Nirvana Swamy Kudlur , Nashwan Abdulqadir J. , Chandra Prem , Khan Abdul Waheed , Alabdulla Majid
Published: August 2021

Background

Intensive care unit (ICU) staff have faced unprecedented challenges during the coronavirus disease 2019 (COVID-19) pandemic, which could significantly affect their mental health and well-being. The present study aimed to investigate perceived stress and post-traumatic stress disorder (PTSD) symptoms reported by ICU staff working directly with COVID-19 patients.

Methods

The Perceived Stress Scale was used to assess perceived stress, the PTSD Diagnostic Scale for the Diagnostic and Statistical Manual of Mental Disorders (5th edition) was used to determine PTSD symptoms, and a sociodemographic questionnaire was used to record different sociodemographic variables.

Results

Altogether, 124 participants (57.2% of whom were men) were included in the analysis. The majority of participants perceived working in the ICU with COVID-19 patients as moderately to severely stressful. Moreover, 71.4% of doctors and 74.4% of nurses experienced moderate-to-severe perceived stress. The staff with previous ICU experience were less likely to have a probable diagnosis of PTSD than those without previous ICU experience.

Conclusions

Assessing perceived stress levels and PTSD among ICU staff may enhance our understanding of COVID-19-induced mental health challenges. Specific strategies to enhance ICU staff’s mental well-being during the COVID-19 pandemic should be employed and monitored regularly. Interventions aimed at alleviating sources of anxiety in a high-stress environment may reduce the likelihood of developing PTSD.