bluuebunny/crossref_metadata_2025 huggingface dataset:
A Flexible Network of Lipid Droplet Associated Proteins Support Embryonic Integrity of C. elegans
Authors: Cao Zhe , Fung Chun Wing , Mak Ho Yi
Published: April 2022

In addition to coordinating the storage and mobilization of neutral fat, lipid droplets (LDs) are conserved organelles that can accommodate additional cargos in order to support animal development. However, it is unclear if each type of cargo is matched with a specific subset of LDs. Here, we report that SEIP-1/seipin defines a subset of oocyte LDs that are required for proper eggshell formation in C. elegans. Using a photoconvertible fluorescent protein-based imaging assay, we found that SEIP-1 positive LDs were selectively depleted after fertilization, coincident of the formation of a lipid-rich permeability barrier of the eggshell. Loss of SEIP-1 function caused impenetrant embryonic arrest, which could be worsened by FAT-3/fatty acyl-CoA desaturase deficiency or suppressed by PLIN-1/Perilipin deficiency. The embryonic development of seip-1; plin-1 mutant in turn depended on the recruitment of RAB-18/Rab18 to LDs, which was not observed in wild type embryos. We propose that SEIP-1 dependent and independent mechanisms act in parallel to ensure the packaging and export of lipid-rich permeability barrier constituents, which involve LDs. The identity of these LDs, as defined by their associated proteins, exhibits unexpected plasticity that ultimately ensures the survival of embryos ex utero.
USP13: Multiple Functions and Target Inhibition
Authors: Li Xiaolong , Yang Ge , Zhang Wenyao , Qin Biying , Ye Zifan , Shi Huijing , Zhao Xinmeng , Chen Yihang , Song Bowei , Mei Ziqing , Zhao Qi , Wang Feng
Published: April 2022

As a deubiquitination (DUB) enzyme, ubiquitin-specific protease 13 (USP13) is involved in a myriad of cellular processes, such as mitochondrial energy metabolism, autophagy, DNA damage response, and endoplasmic reticulum-associated degradation (ERAD), by regulating the deubiquitination of diverse key substrate proteins. Thus, dysregulation of USP13 can give rise to the occurrence and development of plenty of diseases, in particular malignant tumors. Given its implications in the stabilization of disease-related proteins and oncology targets, considerable efforts have been committed to the discovery of inhibitors targeting USP13. Here, we summarize an overview of the recent advances of the structure, function of USP13, and its relations to diseases, as well as discovery and development of inhibitors, aiming to provide the theoretical basis for investigation of the molecular mechanism of USP13 action and further development of more potent druggable inhibitors.
Advancing a Human Onchocerciasis Vaccine From Antigen Discovery to Efficacy Studies Against Natural Infection of Cattle With Onchocerca ochengi
Authors: Zhan Bin , Bottazzi Maria Elena , Hotez Peter J. , Lustigman Sara
Published: April 2022

Human onchocerciasis is a devastating neglected tropical disease caused by infection of the filarial nematodeOnchocerca volvulus. The infection can cause irreversible visual impairment or blindness and stigmatizing dermatitis. More than 32 million people were estimated to be infected withO. volvulusin Africa, and 385,000 suffered from blindness. Even though the implementation of mass drug administration (MDA) with ivermectin has reduced the global prevalence of onchocerciasis,O. volvulusinfection remains challenging to control because MDA with ivermectin cannot be implemented in endemic areas co-endemic with loiasis due to the risk of severe adverse events. There is also emerging drug resistance to ivermectin that further complicates the elimination of onchocerciasis. Thus, the development of a vaccine that would induce protective immunity and reduce infection burden is essential. Efforts to develop prophylactic and/or therapeutic vaccines for onchocerciasis have been explored since the late 1980s by many researchers and entities, and here we summarize the recent advances made in the development of vaccines against the infection ofO. volvulusand onchocerciasis.
Quantitative Flow Ratio or Angiography for the Assessment of Non-culprit Lesions in Acute Coronary Syndromes: Protocol of the Randomized Trial QUOMODO
Authors: Ullrich Helen , Olschewski Maximilian , Belhadj Khelifa-Anis , Münzel Thomas , Gori Tommaso
Published: April 2022

Background

Approximately 50% of the patients undergoing percutaneous coronary intervention (PCI) for acute coronary syndromes (ACS) have additional stenotic lesions in non-infarct-related coronary arteries. The decision whether these stenoses require further treatment is routinely based on angiography alone. The quantitative flow ratio (QFR) is a simple non-invasive method that may help quantify the functional significance of these intermediate coronary artery lesions. The aim of our single-center, randomized superiority trial is to test the impact and efficacy of QFR, as compared to angiography, in the treatment of patients with ACS with multivessel coronary artery disease. Primary goal of the study is to investigate 1. The impact of QFR on the proportion of patients receiving PCI vs. conservative therapy and 2. whether QFR improves angina pectoris and overall cardiovascular outcomes.

Methods and Analysis

After treatment of the culprit lesion(s), a total of 200 consecutive ACS patients will be randomized 1:1 to angiography- vs. QFR-guided revascularization of non-culprit stenoses. Patients and clinicians responsible are blinded to the randomization group. The primary functional endpoint is defined as the proportion of patients assigned to medical treatment in the two groups. The primary clinical endpoint is a composite of death, non-fatal myocardial infarction, revascularization and significant angina at 12 months. Secondary endpoints include changes in the SAQ subgroups, and clinical events at 3- and 12-month follow-up.

Discussion

This study is designed to investigate whether QFR-based decision-making is associated with a decrease in angina and an improved prognosis in patients with multivessel disease.

Trial Registration Number

ClinicalTrials.gov Registry (NCT04808310).
Pre-operative Considerations in Adult Mucopolysaccharidosis Patients Planned for Cardiac Intervention
Authors: Cross Benjamin , Stepien Karolina M. , Gadepalli Chaitanya , Kharabish Ahmed , Woolfson Peter , Tol Govind , Jenkins Petra
Published: April 2022

Mucopolysaccharidoses (MPS) are rare lysosomal storage diseases characterized by multiorgan involvement and shortened longevity. Due to advances in therapies such as enzyme replacement therapy and haematopoietic stem cell therapy, life expectancy has increased posing newer challenges to patients and health professionals. One such challenge is cardiovascular manifestations of MPS, which can be life limiting and cause reduction in quality of life. Any cardiovascular intervention mandates comprehensive, multi-systemic work-up by specialist teams to optimize outcome. We highlight the importance of multidisciplinary evaluation of adult MPS patients requiring cardiovascular intervention. Clinical assessments and investigations are discussed, with a focus on the cardiac, anesthetic, airway, respiratory, radiological and psychosocial factors.
Comparison of Test Methods for Determining the Tensile Strength of Soil and Weak Rocks
Authors: Wang Rong , Li Yanrong , Lv Dongdong , Zhao Weichao , Zhang Chaobo , Zachert Hauke , Eichhoff Gerald , Beroya-Eitner Mary Antonette
Published: April 2022

Tensile strength is a key parameter governing tensile cracking and subsequent failure of soil or rock mass. Existing methods for measuring tensile strength are mainly designed for hard materials and come with inherent problems. As such, they are continuously being adapted and improved by the scientific community. In line with this effort, we recently developed two new tensile test methods for application to soil and weak rocks, namely, the inner hole fracturing test (IHFT) and horizontal compression test (HCT). In this study, we compared the performance of these newly developed methods and the three most commonly used approaches for tensile strength determination, namely, the uniaxial direct tensile test (UDTT), Brazilian test (BT) and three-point bending test (TPBT). Results show that IHFT and HCT exhibit distinct advantages over the three conventional methods when testing soil and weak rocks: first, IHFT and HCT can overcome the eccentric force problem that is a main challenge in UDTT and BT, and second, results obtained from these tests are highly reproducible and stable. Between IHFT and HCT, the latter is found more suitable for routine laboratory testing because of simpler and easier procedure, more stable and reliable results and uniform stress distribution within specimens.
Epidemiology, Treatment and Prognosis Analysis of Small Cell Breast Carcinoma: A Population-Based Study
Authors: Zhu Jiahao , Wu Gang , Zhao Yutian , Yang Bo , Chen Qingqing , Jiang Jianwei , Meng You , Ji Shengjun , Gu Ke
Published: April 2022

Background

Primary small cell breast carcinoma (SCBC) is an uncommon malignancy with highly invasive behavior. The aim of this study was to find out more about the incidence, clinicopathologic characteristics and identify potential prognostic factors of SCBC.

Methods

Data of patients with primary diagnosis of SCBC between 1975 and 2018 were extracted from the Surveillance, Epidemiology, and End Results (SEER) database. The incidence after adjustment for age and percentage change per year in incidence were calculated. Disease-specific survival (DSS) and overall survival (OS) were analyzed among these SCBC patients identified from the SEER database. The whole cohorts were randomized into training and validation cohorts as ratio of 7: 3. Cox regression analysis was performed to determine predictors of survival with the training cohorts. Predictive models were constructed with training cohorts, and nomogram validation was performed using receiver operating characteristic curves, concordance indices and calibration curves in both training and validation cohorts.

Results

323 SCBC patients were enrolled finally during the research period. The overall incidence after adjustment for age between 1990 and 2018 was 0.14 per million per year, and the prevalence of the incidence has plateaued. Most of these tumors were poorly differentiated or undifferentiated. The most prevalent presenting stage was Stage II. Patients identified in this study were randomly divided into training (n = 226) and testing (n = 97) cohorts. Multivariate Cox proportional hazards model showed that chemotherapy, surgery and stage were important predictors of DSS and OS.

Conclusion

SCBC is considered an infrequent breast neoplasm with aggressive characteristics. Tumor stage is associated with poor prognosis. Combination of surgery and chemotherapy is the main treatment for SCBC.
Spermatogonial Stem Cell-Based Therapies: Taking Preclinical Research to the Next Level
Authors: Sanou Iris , van Maaren Jillis , Eliveld Jitske , Lei Qijing , Meißner Andreas , de Melker Annemieke A. , Hamer Geert , van Pelt Ans M. M. , Mulder Callista L.
Published: April 2022

Fertility preservation via biobanking of testicular tissue retrieved from testicular biopsies is now generally recommended for boys who need to undergo gonadotoxic treatment prior to the onset of puberty, as a source of spermatogonial stem cells (SSCs). SSCs have the potential of forming spermatids and may be used for therapeutic fertility approaches later in life. Although in the past 30 years many milestones have been reached to work towards SSC-based fertility restoration therapies, including transplantation of SSCs, grafting of testicular tissue and various in vitro and ex vivo spermatogenesis approaches, unfortunately, all these fertility therapies are still in a preclinical phase and not yet available for patients who have become infertile because of their treatment during childhood. Therefore, it is now time to take the preclinical research towards SSC-based therapy to the next level to resolve major issues that impede clinical implementation. This review gives an outline of the state of the art of the effectiveness and safety of fertility preservation and SSC-based therapies and addresses the hurdles that need to be taken for optimal progression towards actual clinical implementation of safe and effective SSC-based fertility treatments in the near future.
Subsidized or Not, the Impact of Firm Internationalization on Green Innovation—Based on a Dynamic Panel Threshold Model
Authors: Chen Pengyu
Published: April 2022

To alleviate the increasingly serious environmental problems, the environmental governance of relevant firms has received widespread attention. In this paper, based on panel data of Chinese listed firms from 2010–2019, we use the dynamic panel model to verify the non-linear relationship between internationalization and green innovation performance. The dynamic panel threshold model is also constructed to estimate the threshold effect of subsidies between internationalization and green innovation performance. The results show that there is a “U” relationship between internationalization and green innovation. Subsidies can help firms cross the inflection point earlier, and internationalization positively affects green innovation output only when the subsidy exceeds the threshold (16.994). Considering the heterogeneity issue, our study finds that the subsidy threshold for internationalization is bigger for state-owned, non-coastal enterprises, and enterprises with environmental information disclosure compared to other enterprises. In addition, when across the subsidy threshold, state-owned, non-coastal enterprises, and enterprises without environmental information disclosure are better able to stimulate green innovation output. This provides evidence and policy directions for other emerging developing countries.
Phase-Resolved Lockdown Features of Pollution Parameters Over an Urban and Adjoining Rural Region During COVID-19
Authors: Sonbawne Sunil M. , Fadnavis Suvarna , Vijayakumar K. , Devara Panuganti C. S. , Chavan Prashant
Published: April 2022

In this study, we investigate the temporal variations in columnar aerosol pollutants and their possible association with the simultaneously measured black carbon (BC) aerosol mass concentration and associated biomass burning (BB) over urban (Delhi) and rural (Panchgaon) sites during the lockdown phases of the COVID-19 pandemic. We also show the impact of lockdown measures on boundary layer ozone and its primary precursors, NO2, and water vapor (H2O), potent greenhouse gases that destroy protective ozone. For this purpose, we used multiple datasets, namely, black carbon (BC) aerosol mass concentration and biomass burning (BB) aerosols using an aethalometer at Amity University Haryana (AUH), Panchgaon, India, and satellite retrievals from NASA’s MODIS and OMI at both the stations. The analysis was conducted during the pre-lockdown period (1–25 March), lockdown 1st phase (25 March–14 April), lockdown 2nd phase (15 April–3 May), lockdown 3rd phase (4–17 May), lockdown 4th phase (18–31 May), and post-lockdown (1–30 June) period in 2020. Our diagnostic analysis shows a substantial reduction in AOD (Delhi: −20% to −80%, Panchgaon: −20% to −80%) and NO2 (Delhi: −10% to −42.03%, Panchgaon −10% to −46.54%) in comparison with climatology (2010–2019) during all four phases of lockdown. The reduction in AOD is attributed to lockdown measures and less transport of dust from west Asia than climatology. Despite a reduction in NO2, there is an increase in the ozone amount (Delhi: 1% to 8% and Panchgaon: 1% to 10%) during lockdown I, II, and III phases. The observed enhancement in ozone may be resultant from the complex photochemical processes that involve the presence of NO2, CO, volatile organic compounds (VOCs), and water vapor. The reduction in AOD and NO2 and enhancement in ozone are stronger at the rural site, Panchgaon than that at the urban site, Delhi.
Evaluation of Sea Ice Concentration Data Using Dual-Polarized Ratio Algorithm in Comparison With Other Satellite Passive Microwave Sea Ice Concentration Data Sets and Ship-Based Visual Observations
Authors: Zong Fangyi , Zhang Shugang , Chen Ping , Yang Lipeng , Shao Qiuli , Zhao Jinping , Wei Lai
Published: April 2022

The dual-polarized ratio (DPR) algorithm is a new algorithm that enable calculation of Arctic sea ice concentration from the 36.5-GHz channel of the sensor Advanced Microwave Scanning Radiometer for EOS/Advanced Microwave Scanning Radiometer 2 (AMSR-E/AMSR2). In this paper, we demonstrate results that the sea ice concentration data using DPR algorithm (DPR-AMSR) are evaluated and compared with other eight Arctic sea ice concentration data products with respect to differences in sea ice concentration, sea ice area, and sea ice extent. On a pan-Arctic scale, the evaluation results are mostly very similar between DPR-AMSR and the bootstrap algorithm from AMSR-E/AMSR2 (BT-AMSR), the bootstrap algorithm from SSM/I or SSMIS (BT-SSMI), the ARTIST Sea Ice algorithm from AMSR-E/AMSR2 (ASI-AMSR), and the enhanced NASA Team algorithm from AMSR-E/AMSR2 (NT2-AMSR). Among of these products, the differences in sea ice concentration agree within ±5%. However, European Space Agency Climate Change Initiative algorithm from AMSR-E/AMSR2 (SICCI-AMSR), the European Organisation for the Exploitation of Meteorological Satellites Ocean and Sea Ice Satellite Application Facility from SSM/I or SSMIS (OSI-SSMI), the ARTIST Sea Ice algorithm from SSM/I or SSMIS (ASI-SSMI), and the NASA Team algorithm from SSM/I or SSMIS (NT1-SSMI) are all lower than DPR-AMSR at sea ice edge. And NT1-SSMI had the largest negative difference, which was lower than -15% or even 20%.The difference of sea ice area was consistently within ±0.5 million km2 between DPR-AMSR and BT-AMSR, BT-SSMI, ASI-AMSR, and NT2-AMSR in all years. The smallest difference was with BT-SSMI (less than 0.1 million km2), whereas the largest difference was with NT1-SSMI (up to 1.5 million km2). In comparisons of sea ice extent, BT-AMSR, NT1-SSMI, and NT2-AMSR estimates were consistent with that of DPR-AMSR and were within ±0.5 million km2. However, differences exceeded 0.5 million km2 between DPR-AMSR and the other data sets. When ship-based visual observation (OBS) values ranged from 85% to 100%, the difference between DPR-AMSR and OBS was less than 1%. There were relatively large differences between DPR-AMSR and OBS when OBS values were less than 85% or were recorded during the summer, although those differences were also within 10%.
Soil Nitrous Oxide Emission and Methane Exchange From Diversified Cropping Systems in Pannonian Region
Authors: Hüppi Roman , Horváth László , Dezső József , Puhl-Rezsek Marietta , Six Johan
Published: April 2022

Diversified farming systems are promoted to improve ecosystem services in agriculture while maintaining productivity. Intercropping could improve soil quality, the stability of yields and climate resilience. Whether direct emissions of greenhouse gases from soil are reduced as well, depends on the specific measures of diversification. Here, we determined the greenhouse gas emissions from soils of two diversification experiments in the Pannonian climate of Hungary. Firstly, in an asparagus field, oat and field pea was introduced as intercrop between the asparagus berms. Secondly, grass and aromatic herbs were intercropped in a vineyard between the grape rows. The results show that especially for nitrous oxide, average treatment emissions can increase with additional legumes (+252% with intercropped field peas) but decrease with aromatic herbs (−66%). No significant changes were found for methane exchange. This shows that, while other ecosystem services can be increased by intercropping, changes in soil greenhouse gas emissions by intercropping are highly context dependent.
Bioaccumulation and Risk Assessment of Potentially Toxic Elements in Soil-Rice System in Karst Area, Southwest China
Authors: Zhang Chunlai , Zou Xia , Yang Hui , Liang Jianhong , Zhu Tongbin
Published: April 2022

The accumulation of potentially toxic elements (PTE) in a soil–rice system poses a significant issue of concern in agricultural soils, particularly in the polluted or high PTE geological background regions, such as karst areas. The source identification, bioaccumulation factors of PTE, and its health risk were investigated by correlation analysis, principal components analysis, and single/comprehensive assessments in a soil-rice system in Mashan County, Guangxi Province. The results showed that the mean contents of PTE in rice rhizosphere soil samples were higher than Guangxi surface soil, but lower than Mashan background. Of the samples, 84.21% have Cd content exceeding the soil environmental quality -- risk control standard for soil contamination of agricultural land in China (GB 15618-2018) risk screening value. The Nemerow comprehensive pollution index indicated that 21.05 and 26.32% of the soil samples were moderately and heavily polluted. The contents of pH-related exchangeable Ca, exchangeable Mg, and redox-related available Fe and available Mn in soil affected the bioaccumulation of PTE in rice. In all the rice samples, 55.26% of Cd and 31.58% of Pb exceed the maximum allowable value of contaminants in rice recommended by the national food safety standard for maximum levels of contaminants in foods in China (GB 2762-2017). The average targeted hazard quotient values (THQ) of PTE decreased in an order of As > Cd > Cr > Cu > Zn > Pb > Hg, and the degree of health risk it posed to the population was Children > Female > Male. The hazard index (HI) of all samples was greater than one due to all THQAs and the THQCd of more than half samples were above 1, which implied that the residents were exposed to non-carcinogenic risk by rice ingestion. Therefore, the PTE in the karst area with a high geological background can be absorbed and migrated by crops, leading to a greater health risk to humans, which should be paid attention to in future research and agricultural management.
Climate-Driven Legacies in Simulated Microbial Communities Alter Litter Decomposition Rates
Authors: Wang Bin , Allison Steven D.
Published: April 2022

The mechanisms underlying diversity-functioning relationships have been a consistent area of inquiry in biogeochemistry since the 1950s. Though these mechanisms remain unresolved in soil microbiomes, many approaches at varying scales have pointed to the same notion—composition matters. Confronting the methodological challenge arising from the complexity of microbiomes, this study used the model DEMENTpy, a trait-based modeling framework, to explore trait-based drivers of microbiome-dependent litter decomposition. We parameterized DEMENTpy for five sites along a climate gradient in Southern California, United States, and conducted reciprocal transplant simulations analogous to a prior empirical study. The simulations demonstrated climate-dependent legacy effects of microbial communities on plant litter decomposition across the gradient. This result is consistent with the previous empirical study across the same gradient. An analysis of community-level traits further suggests that a 3-way tradeoff among resource acquisition, stress tolerance, and yield strategies influences community assembly. Simulated litter decomposition was predictable with two community traits (indicative of two of the three strategies) plus local environment, regardless of the system state (transient vs. equilibrium). Although more empirical confirmation is still needed, community traits plus local environmental factors (e.g., environment and litter chemistry) may robustly predict litter decomposition across spatial-temporal scales. In conclusion, this study offers a potential trait-based explanation for climate-dependent community effects on litter decomposition with implications for improved understanding of whole-ecosystem functioning across scales.
Comparing the Effectiveness of Biodiversity Conservation Across Different Regions by Considering Human Efforts
Authors: Dong Kaikai , Liu Zhaoli , Li Ying , Chen Ziqi , Hou Guanglei , Sun Jingkuan
Published: April 2022

The effective allocation of funds is of significant importance for biodiversity conservation, but there is currently no scientific method for comparing the effectiveness of biodiversity conservation across different regions. Existing studies omit differences in the ecological background, such as the terrain, climate, hydrology, soil, and ecosystem, or do not differentiate between the impacts caused by humans and nature. To address these limitations, we take habitat quality as a proxy for biodiversity and quantify the human-induced habitat quality changes as a means of measuring the efforts of management departments, with the background differences eliminated using a reference condition index. The method is applied to the San Jiang Plain Wetlands and Northwest Tibet Qiang Tang Plateau Biodiversity National Key Ecological Function Region in China. The results show that the effects of human activities on habitat improvement or degradation are overestimated or underestimated if there is no differentiation between human and natural causes. Human-induced habitat quality changes broadly reflect the human efforts toward biodiversity conservation. By considering the human efforts and background differentiation, the proposed method allows the effectiveness of biodiversity conservation to be compared across different regions. This study provides a scientific reference for China’s transfer payment policy and for the biodiversity funds allocated in other countries. Furthermore, our results will guide the practice of improving habitat quality and biodiversity.
Soil Fungi Exposed to Warming Temperatures and Shrinking Snowpack in a Northern Hardwood Forest Have Lower Capacity for Growth and Nutrient Cycling
Authors: Finestone Julia , Templer Pamela H. , Bhatnagar Jennifer M.
Published: April 2022

Projections for the northeastern United States indicate that mean air temperatures will rise and snowfall will become less frequent, causing more frequent soil freezing. To test fungal responses to these combined chronic and extreme soil temperature changes, we conducted a laboratory-based common garden experiment with soil fungi that had been subjected to different combinations of growing season soil warming, winter soil freeze/thaw cycles, and ambient conditions for 4 years in the field. We found that fungi originating from field plots experiencing a combination of growing season warming and winter freeze/thaw cycles had inherently lower activity of acid phosphatase, but higher cellulase activity, that could not be reversed in the lab. In addition, fungi quickly adjusted their physiology to freeze/thaw cycles in the laboratory, reducing growth rate, and potentially reducing their carbon use efficiency. Our findings suggest that less than 4 years of new soil temperature conditions in the field can lead to physiological shifts by some soil fungi, as well as irreversible loss or acquisition of extracellular enzyme activity traits by other fungi. These findings could explain field observations of shifting soil carbon and nutrient cycling under simulated climate change.
Genetic Analysis of a Pedigree With Antithrombin and Prothrombin Compound Mutations and Antithrombin Heterozygotes
Authors: Zhang Haiyue , Hu Yiling , Pan Dongli , Xv Yuehua , Shen Weifeng
Published: April 2022

Background and Aims: Antithrombin (AT) is the most important physiological inhibitor in vivo, and coagulation factor II (FII) or prothrombin is a coagulation factor vital to life. The purpose of our research was to illustrate the connection between gene mutations and the corresponding deficiencies of AT and FII.

Methods: Functional and molecular analyses were performed. The possible impact of the mutation was analyzed by online bioinformatics software. ClustalX-2.1-win and PyMol/Swiss-Pdb Viewer software were used for conservative analyses and to generate molecular graphic images, respectively.

Results: The proband showed a lower limb venous thrombosis and acute pulmonary embolism infarction with reduced AT activity (50%). His mother, with subcutaneous ecchymosis, had reduced activities of AT and FII, of 44 and 5%, respectively. Molecular analysis showed that both the proband and his mother carried c.964A > T (p.Lys322stop) heterozygotes in SERPINC1. The difference was that his mother carried homozygous c.494C > T (p.Thr165Met) in F2, while the proband was wild type. Bioinformatics and model analysis indicated that mutations may destroy the function and structure of AT and FII protein.

Conclusion: This study identified a novel mutation of SERPINC1 and a missense mutation of F2, which may be the molecular mechanism leading to AT and FII deficiency in this family. It will help genetic diagnosis and counseling for thrombotic families.
Evaluating the Causal Effects of TIMP-3 on Ischaemic Stroke and Intracerebral Haemorrhage: A Mendelian Randomization Study
Authors: Xiao Linxiao , Zou Xuelun , Liang Yan , Wang Yuxiang , Zeng Lang , Wu Jianhuang
Published: April 2022

Aim: Since tissue inhibitors of matrix metalloproteinase 3 (TIMP-3) was reported to be a potential risk factor of atherosclerosis, aneurysm, hypertension, and post-ischaemic neuronal injury, it may also be a candidate risk factor of stress. Therefore, this study was designed to explore the causal role of TIMP-3 in the risk of ischaemic stroke (IS) and intracerebral haemorrhage (ICH), which are the two main causes of stress via this Mendelian Randomisation (MR) study.

Methods: The summarised data of TIMP-3 level in circulation was acquired from the Cooperative Health Research in the Region of Augsburg public database and the outcome of IS and ICH was obtained from genome-wide association studies conducted by MEGASTROKE and the International Stroke Genetics Consortium, respectively. Five statistical methods including inverse-variance weighting, weighted-median analysis, MR-Egger regression, MR Pleiotropy RESidual Sum and Outlier test, and MR-Robust Adjusted Profile Score were applied to evaluate the causal role of TIMP-3 in the occurrence of IS and ICH. Inverse-variance weighting was applied for assessing causality. Furthermore, heterogeneity and pleiotropic tests were utilised to confirm the reliability of this study.

Results: We found that TIMP-3 could be a positively causal relationship with the incidence of IS (OR = 1.026, 95% CI: 1.007–1.046, p = 0.0067), especially for the occurrence of small vessel stroke (SVS; OR = 1.045, 95% CI: 1.016–1.076, p = 0.0024). However, the causal effects of TIMP-3 on another IS subtype cardioembolic stroke (CES; OR = 1.049, 95% CI: 1.006–1.094, p = 0.024), large artery stroke (LAS; OR = 1.0027, 95% CI: 0.9755–1.0306, p = 0.849) and ICH (OR = 0.9900, 95% CI: 0.9403–1.0423, p = 0.701), as well as ICH subtypes were not observed after Bonferroni corrections (p = 0.00714).

Conclusion: Our results revealed that high levels of circulating TIMP-3 causally increased the risk of developing IS and SVS, but not CES, LAS, ICH, and all ICH subtypes. Further investigation is required to elucidate the underlying mechanism.
Identification of tRNA-Related Fragments and Their Potential Regulatory Effects in Thyroid-Associated Ophthalmopathy
Authors: Yue Zifan , Tong Fei , Zeng Chengcheng , Wei Ruili
Published: April 2022

Recently, the potential role of tRNA-related fragments (tRFs) in ophthalmic diseases has been extensively researched. However, systematic studies on the potential regulatory effects of tRFs in thyroid-associated ophthalmopathy (TAO) are lacking. We used high-throughput sequencing techniques to measure expression levels of mRNAs and tRFs in patients with TAO, and the results were verified by real-time quantitative reverse transcription polymerase chain reaction (q-PCR). Next, the potential biological regulatory effect of differentially expressed tRFs was analyzed, and potential downstream target RNAs of differentially expressed tRFs were predicted to explore the potential role of tRFs as therapeutic targets and biomarkers of TAO. A total of 50 tRFs and 361 mRNAs were dysregulated in the TAO group, and tRF5-GluCTC, PMAIP1, HSD17B2 and ATF3 were verified to be significantly differentially expressed in TAO. Our research reveals that several associated pathways likely play a role in the pathogenesis of TAO. By targeting ATF3, HSD17B2 and PMAIP1, tRF5-GluCTC may play a potential role in regulating the orbital fibroblast adipogenic response and fibrotic hyperplasia in patients with TAO.
Early B Cell and Plasma Cell Kinetics Upon Treatment Initiation Portend Flares in Systemic Lupus Erythematosus: A Post-Hoc Analysis of Three Phase III Clinical Trials of Belimumab
Authors: Parodis Ioannis , Gomez Alvaro , Chow Jun Weng , Borg Alexander , Lindblom Julius , Gatto Mariele
Published: April 2022

Objective

To investigate changes in B cell subsets in relation to disease flares upon initiation of standard therapy (ST) plus belimumab or placebo in patients with systemic lupus erythematosus (SLE).

Patients and Methods

Using data from the BLISS-76, BLISS-SC and BLISS Northeast Asia trials, we investigated associations of relative to baseline rapid (through week 8) and early (through week 24) changes in peripheral B cell subsets, anti-dsDNA and complement levels with the occurrence of disease flares from week 24 through week 52 (Mann-Whitney U tests) or the entire study follow-up (Cox regression analysis), assessed using the SELENA-SLEDAI Flare Index.

Results

Patients on ST alone who flared displayed less prominent early decreases in CD19+CD20-CD138+ long-lived plasma cells (-16.1% versus -35.1%; P=0.012). In all arms combined, patients who developed severe flares showed less prominent early decreases in CD19+CD20-CD138+ long-lived plasma cells (-23.5% versus -39.4%; P=0.028) and CD19+CD27brightCD38bright SLE-associated plasma cells (-19.0% versus -27.8%; P=0.045). After adjustment for rapid changes, early increases in overall CD19+CD20+ B cells (HR: 1.81; 95% CI: 1.08–3.05; P=0.024) and early increases or no return after a rapid expansion in CD19+CD20+CD27+ memory B cells (HR: 1.58; 95% CI: 1.18–2.11; P=0.002) portended subsequent severe flares. Patients who developed flares of any severity showed no or less prominent rapid (0.0% versus -12.5%; P<0.001) or early (-1.9% versus -21.7%; P<0.001) decreases in anti-dsDNA levels, and patients who developed severe flares showed no or less prominent early decreases in anti-dsDNA levels (0.0% versus -13.3%; P=0.020). Changes in complement levels exhibited no ability to distinguish flaring from non-flaring patients.

Conclusions

Increase or lack of decrease in certain circulating B cell subsets or anti-dsDNA levels upon treatment initiation for active SLE heralded subsequent severe disease flares. A rapid expansion of memory B cells may signify sustained response to therapy when followed by a subsequent drop, while no return or delayed increases in memory B cells may portend flaring. Peripheral B cell and serological marker kinetics may help identify patients in whom therapeutic modifications could protect against flare development, and may hence prove a useful complement to traditional surveillance and early treatment evaluation in SLE.
Genetic Surveillance of Five SARS-CoV-2 Clinical Samples in Henan Province Using Nanopore Sequencing
Authors: Wang Yanan , Chen Duo , Zhu Chaofeng , Zhao Zhenhua , Gao Shanshan , Gou Jianjun , Guo Yongjun , Kong Xiangdong
Published: April 2022

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly spread and poses a major threat to public health worldwide. The whole genome sequencing plays a crucial role in virus surveillance and evolutionary analysis. In this study, five genome sequences of SARS-CoV-2 were obtained from nasopharyngeal swab samples from Zhengzhou, China. Following RNA extraction and cDNA synthesis, multiplex PCR was performed with two primer pools to produce the overlapped amplicons of ~1,200 bp. The viral genomes were obtained with 96% coverage using nanopore sequencing. Forty-five missense nucleotide mutations were identified; out of these, 5 mutations located at Nsp2, Nsp3, Nsp14, and ORF10 genes occurred with a <0.1% frequency in the global dataset. On the basis of mutation profiles, five genomes were clustered into two sublineages (B.1.617.2 and AY.31) or subclades (21A and 21I). The phylogenetic analysis of viral genomes from several regions of China and Myanmar revealed that five patients had different viral transmission chains. Taken together, we established a nanopore sequencing platform for genetic surveillance of SARS-CoV-2 and identified the variants circulating in Zhengzhou during August 2021. Our study provided crucial support for government policymaking and prevention and control of COVID-19.
AMG853, A Bispecific Prostaglandin D2 Receptor 1 and 2 Antagonist, Dampens Basophil Activation and Related Lupus-Like Nephritis Activity in Lyn-Deficient Mice
Authors: Pellefigues Christophe , Tchen John , Saji Chaimae , Lamri Yasmine , Charles Nicolas
Published: April 2022

Systemic lupus erythematosus is a complex autoimmune disease during which patients develop autoantibodies raised against nuclear antigens. During the course of the disease, by accumulating in secondary lymphoid organs (SLOs), basophils support autoreactive plasma cells to amplify autoantibody production. We have recently shown that murine lupus-like disease could be controlled by 10 days of oral treatment with a combination of prostaglandin D2 (PGD2) receptor (PTGDR) antagonists through the inhibition of basophil activation and recruitment to SLOs. Importantly, inhibiting solely PTGDR-1 or PTGDR-2 was ineffective, and the development of lupus-like disease could only be dampened by using antagonists for both PTGDR-1 and PTGDR-2. Here, we aimed at establishing a proof of concept that a clinically relevant bispecific antagonist of PTGDR-1 and PTGDR-2 could be efficient to treat murine lupus-like nephritis. Diseased Lyn-deficient female mice received treatment with AMG853 (vidupiprant, a bispecific PTGDR-1/PTGDR-2 antagonist) for 10 days. This led to the dampening of basophil activation and recruitment in SLOs and was associated with a decrease in plasmablast expansion and immunoglobulin E (IgE) production. Ten days of treatment with AMG853 was consequently sufficient in reducing the dsDNA-specific IgG titers, circulating immune complex glomerular deposition, and renal inflammation, which are hallmarks of lupus-like disease. Thus, bispecific PTGDR-1 and PTGDR-2 antagonists, such as AMG853, are a promising class of drugs for the treatment or prevention of organ damage in systemic lupus erythematosus.
Conservation and Evolution of Antigenic Determinants of SARS-CoV-2: An Insight for Immune Escape and Vaccine Design
Authors: Jaiswal Varun , Lee Hae-Jeung
Published: April 2022

Coronavirus disease 2019 (COVID-19) is the most devastating pandemic of the century, which is still far from over. The remarkable success of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines is the working hope, but the evolving variants are the huge concern that can turn the tide. Potential immune escape mutations (PIEMs) in the past and circulating variants were not studied at large scale (all available data). Hence, the conservation of antigenic determinants (epitopes) was analyzed in all available sequences of SARS-CoV-2 according to time (months), proteins, hosts, and variants. Numerous highly conserved B- and T-cell epitopes were identified in 24 proteins of SARS-CoV-2. A decrease in the conservation of epitopes with time was observed in almost all proteins, which was more rapid in neutralizing epitopes. Delta variant still has the highest PIEM in the circulating strains, which pose threat to the effectiveness of current vaccines. The inclusion of identified, highly conserved, and important epitopes in subunit vaccines can increase vaccine effectiveness against evolving variants. Trends in the conservation of epitopes in different proteins, hosts, and variants with time may also help to inspire the counter measure against the current pandemic.
Effect of Factor H on Complement Alternative Pathway Activation in Human Serum Remains on Porcine Cells Lacking N-Glycolylneuraminic Acid
Authors: Lee Haneulnari , Park Eun Mi , Ko Nayoung , Choi Kimyung , Oh Keon Bong , Kang Hee Jung
Published: April 2022

Background

Triple knockout (TKO) donor pigs lacking alpha-1,3-galactose (Gal),N-glycolylneuraminic acid (Neu5Gc), and Sd(a) expressions were developed to improve the clinical success of xenotransplantation. Neu5Gc, a sialic acid expressed on cell surfaces, recruits factor H to protect cells from attack by the complement system. Lack of Neu5Gc expression may cause unwanted complement activation, abrogating the potential benefit of gene-modified donor pigs. To investigate whether TKO porcine cells display increased susceptibility to complement activation in human serum, pathway-specific complement activation, apoptosis, and human platelet aggregation by porcine cells were compared betweenalpha-1,3-galactosyltransferasegene-knockout (GTKO) and TKO porcine cells.

Methods

Primary porcine peripheral blood mononuclear cells (pPBMCs) and endothelial cells (pECs) fromGTKO and TKO pigs were used. Cells were incubated in human serum diluted in gelatin veronal buffer (GVB++) or Mg++-EGTA GVB, and C3 deposition and apoptotic changes in these cells were measured by flow cytometry. C3 deposition levels were also measured after incubating these cells in 10% human serum supplemented with human factor H. Platelet aggregation in human platelet-rich plasma containingGTKO or TKO pECs was analyzed.

Results

The C3 deposition level inGTKO pPBMCs or pECs in GVB++was significantly higher than that of TKO pPBMCs or pECs, respectively, but C3 deposition levels in Mg++-EGTA-GVB were comparable between them. The addition of factor H into the porcine cell suspension in 10% serum in Mg++-EGTA-GVB inhibited C3 deposition in a dose-dependent manner, and the extent of inhibition by factor H was similar betweenGTKO and TKO porcine cells. The percentage of late apoptotic cells in porcine cell suspension in GVB++increased with the addition of human serum, of which the net increase was significantly less in TKO pPBMCs than inGTKO pPBMCs. Finally, the lag time of platelet aggregation in recalcified human plasma was significantly prolonged in the presence of TKO pECs compared to that in the presence ofGTKO pECs.

Conclusion

TKO genetic modification protects porcine cells from serum-induced complement activation and apoptotic changes, and delays recalcification-induced human platelet aggregation. It does not hamper factor H recruitment on cell surfaces, allowing the suppression of alternative complement pathway activation.
Signaling Pathways That Mediate Alveolar Macrophage Activation by Surfactant Protein A and IL-4
Authors: García-Fojeda Belén , Minutti Carlos M. , Montero-Fernández Carlos , Stamme Cordula , Casals Cristina
Published: April 2022

Activation of tissue repair program in macrophages requires the integration of IL-4/IL-13 cytokines and tissue-specific signals. In the lung, surfactant protein A (SP-A) is a tissue factor that amplifies IL-4Rα-dependent alternative activation and proliferation of alveolar macrophages (AMs) through the myosin18A receptor. However, the mechanism by which SP-A and IL-4 synergistically increase activation and proliferation of AMs is unknown. Here we show that SP-A amplifies IL-4-mediated phosphorylation of STAT6 and Akt by binding to myosin18A. Blocking PI3K activity or the myosin18A receptor abrogates SP-A´s amplifying effects on IL-4 signaling. SP-A alone activates Akt, mTORC1, and PKCζ and inactivates GSK3α/β by phosphorylation, but it cannot activate arginase-1 activity or AM proliferation on its own. The combined effects of IL-4 and SP-A on the mTORC1 and GSK3 branches of PI3K-Akt signaling contribute to increased AM proliferation and alternative activation, as revealed by pharmacological inhibition of Akt (inhibitor VIII) and mTORC1 (rapamycin and torin). On the other hand, the IL-4+SP-A-driven PKCζ signaling axis appears to intersect PI3K activation with STAT6 phosphorylation to achieve more efficient alternative activation of AMs. Consistent with IL-4+SP-A-driven activation of mTORC1 and mTORC2, both agonists synergistically increased mitochondrial respiration and glycolysis in AMs, which are necessary for production of energy and metabolic intermediates for proliferation and alternative activation. We conclude that SP-A signaling in AMs activates PI3K-dependent branched pathways that amplify IL-4 actions on cell proliferation and the acquisition of AM effector functions.
Platelet-Depletion of Whole Blood Reveals That Platelets Potentiate the Release of IL-8 From Leukocytes Into Plasma in a Thrombin-Dependent Manner
Authors: Quach Huy Quang , Johnson Christina , Ekholt Karin , Islam Rakibul , Mollnes Tom Eirik , Nilsson Per H.
Published: April 2022

Objective

In a recent study, we found an elevated level of interleukin 8 (IL-8) in response to bacterial incubation in thrombin-sufficient human whole blood anticoagulated by the fibrin polymerization blocking peptide GPRP. Whether thrombin directly activated leukocytes or mediated the release via thrombin-dependent activation of platelets remains unresolved. Herein, we addressed the role of thrombin and platelets in IL-8 release.

Methods

We separated platelets from whole blood using a combination of 0.7% (w/v) citrate and GPRP for attenuating the hemostatic response during the separation of platelets. Cytokine responses were compared in whole blood and platelet-depleted blood upon Escherichia coli incubation. Cytokine responses were also profiled with and without reconstitution of either platelets or the supernatant from activated platelets.

Results

Platelets were not activated during the separation process but responded to stimuli upon re-calcification. Plasma levels of IL-1β, IL-1Ra, IL-6, IL-8, IP-10, MIP-1α, and MIP-1β were significantly reduced in platelet-depleted blood compared to whole blood, but recovered in the presence of platelets, or with the supernatant of activated platelets. The leukocyte fraction and platelets were each found to contribute to the elevation of IL-8 at around 5 ng/ml; however, if combined, the release of IL-8 increased to 26 ng/ml. This process was dependent on thrombin since the levels of IL-8 remained at 5 ng/ml in whole blood if thrombin was blocked. Intracellular staining revealed that monocytes were the main source for IL-8 expression.

Conclusion

Our findings suggest that the release of IL-8 is mediated by the leukocytes, mainly monocytes, but potentiated via thrombin-dependent activation of platelets.