bluuebunny/crossref_metadata_2025 huggingface dataset:
Validation of Effective Extracellular Vesicles Isolation Methods Adapted to Field Studies in Malaria Endemic Regions
Authors: Zoia Matteo , Yesodha Subramanian Bibin , Eriksson Klara Kristin , Ravi Meera Sruthi , Yaghmaei Shekoofeh , Fellay Isabelle , Scolari Brigitte , Walch Michael , Mantel Pierre-Yves
Published: May 2022

Malaria affects the poorer regions of the world and is of tremendous health and economic burden for developing countries. Extracellular vesicles (EVs) are small vesicles released by almost any cells in the human body, including malaria infected red blood cells. Recent evidence shows that EVs might contribute to the pathogenesis of malaria. In addition, EVs hold considerable value in biomarker discovery. However, there are still significant gaps in our understanding of EV biology. So far most of our knowledge about EVs in malaria comes from in vitro work. More field studies are required to gain insight into their contribution to the disease and pathogenesis under physiological conditions. However, to perform research on EVs in low-income regions might be challenging due to the lack of appropriate equipment to isolate EVs. Therefore, there is a need to develop and validate EV extraction protocols applicable to poorly equipped laboratories. We established and validated two protocols for EV isolation from cell culture supernatants, rodent and human plasma. We compared polyethylene glycol (PEG) and salting out (SA) with sodium acetate for precipitation of EVs. We then characterized the EVs by Transmission Electron Microscopy (TEM), Western Blot, Size-exclusion chromatography (SEC), bead-based flow cytometry and protein quantification. Both protocols resulted in efficient purification of EVs without the need of expensive material or ultracentrifugation. Furthermore, the procedure is easily scalable to work with large and small sample volumes. Here, we propose that both of our approaches can be used in resource limited countries, therefore further helping to close the gap in knowledge of EVs during malaria.
Pancreatic Cancer: Nucleic Acid Drug Discovery and Targeted Therapy
Authors: Dai Hong , Abdullah Razack , Wu Xiaoqiu , Li Fangfei , Ma Yuan , Lu Aiping , Zhang Ge
Published: May 2022

Pancreatic cancer (PC) is one of the most lethal cancers with an almost 10% 5-year survival rate. Because PC is implicated in high heterogeneity, desmoplastic tumor-microenvironment, and inefficient drug-penetration, the chemotherapeutic strategy currently recommended for the treatment of PC has limited clinical benefit. Nucleic acid-based targeting therapies have become strong competitors in the realm of drug discovery and targeted therapy. A vast evidence has demonstrated that antibody-based or alternatively aptamer-based strategy largely contributed to the elevated drug accumulation in tumors with reduced systematic cytotoxicity. This review describes the advanced progress of antisense oligonucleotides (ASOs), small interfering RNAs (siRNAs), microRNAs (miRNAs), messenger RNA (mRNAs), and aptamer-drug conjugates (ApDCs) in the treatment of PC, revealing the bright application and development direction in PC therapy.
Lung Cancer Induces NK Cell Contractility and Cytotoxicity Through Transcription Factor Nuclear Localization
Authors: Wong Darren Chen Pei , Lee E Hui Clarissa , Er Junzhi , Yow Ivan , Koean Ricky Abdi Gunawan , Ang Owen , Xiao Jingwei , Low Boon Chuan , Ding Jeak Ling
Published: May 2022

Actomyosin-mediated cellular contractility is highly conserved for mechanotransduction and signalling. While this phenomenon has been observed in adherent cell models, whether/how contractile forces regulate the function of suspension cells like natural killer (NK) cells during cancer surveillance, is unknown. Here, we demonstrated in coculture settings that the evolutionarily conserved NK cell transcription factor, Eomes, undergoes nuclear shuttling during lung cancer cell surveillance. Biophysical and biochemical analyses revealed mechanistic enhancement of NK cell actomyosin-mediated contractility, which is associated with nuclear flattening, thus enabling nuclear entry of Eomes associated with enhanced NK cytotoxicity. We found that NK cells responded to the presumed immunosuppressive TGFβ in the NK-lung cancer coculture medium to sustain its intracellular contractility through myosin light chain phosphorylation, thereby promoting Eomes nuclear localization. Therefore, our results demonstrate that lung cancer cells provoke NK cell contractility as an early phase activation mechanism and that Eomes is a plausible mechano-responsive protein for increased NK cytotoxicity. There is scope for strategic application of actomyosin-mediated contractility modulating drugsex vivo,to reinvigorate NK cells prior to adoptive cancer immunotherapyin vivo(177 words).
G Protein-Coupled Receptors as Potential Intercellular Communication Mediators in Trypanosomatidae
Authors: Díaz Emilia , Febres Anthony , Giammarresi Michelle , Silva Adrian , Vanegas Oriana , Gomes Carlos , Ponte-Sucre Alicia
Published: May 2022

Detection and transduction of environmental signals, constitute a prerequisite for successful parasite invasion; i.e., Leishmania transmission, survival, pathogenesis and disease manifestation and dissemination, with diverse molecules functioning as inter-cellular signaling ligands. Receptors [i.e., G protein-coupled receptors (GPCRs)] and their associated transduction mechanisms, well conserved through evolution, specialize in this function. However, canonical GPCR-related signal transduction systems have not been described in Leishmania, although orthologs, with reduced domains and function, have been identified in Trypanosomatidae. These inter-cellular communication means seem to be essential for multicellular and unicellular organism’s survival. GPCRs are flexible in their molecular architecture and may interact with the so-called receptor activity-modifying proteins (RAMPs), which modulate their function, changing GPCRs pharmacology, acting as chaperones and regulating signaling and/or trafficking in a receptor-dependent manner. In the skin, vasoactive- and neuro- peptides released in response to the noxious stimuli represented by the insect bite may trigger parasite physiological responses, for example, chemotaxis. For instance, in Leishmania (V.) braziliensis, sensory [Substance P, SP, chemoattractant] and autonomic [Vasoactive Intestinal Peptide, VIP, and Neuropeptide Y, NPY, chemorepellent] neuropeptides at physiological levels stimulate in vitro effects on parasite taxis. VIP and NPY chemotactic effects are impaired by their corresponding receptor antagonists, suggesting that the stimulated responses might be mediated by putative GPCRs (with essential conserved receptor domains); the effect of SP is blocked by [(D-Pro 2, D-Trp7,9]-Substance P (10-6 M)] suggesting that it might be mediated by neurokinin-1 transmembrane receptors. Additionally, vasoactive molecules like Calcitonin Gene-Related Peptide [CGRP] and Adrenomedullin [AM], exert a chemorepellent effect and increase the expression of a 24 kDa band recognized in western blot analysis by (human-)-RAMP-2 antibodies. In-silico search oriented towards GPCRs-like receptors and signaling cascades detected a RAMP-2-aligned sequence corresponding to Leishmania folylpolyglutamate synthase and a RAMP-3 aligned protein, a hypothetical Leishmania protein with yet unknown function, suggesting that in Leishmania, CGRP and AM activities may be modulated by RAMP- (-2) and (-3) homologs. The possible presence of proteins and molecules potentially involved in GPCRs cascades, i.e., RAMPs, signpost conservation of ancient signaling systems associated with responses, fundamental for cell survival, (i.e., taxis and migration) and may constitute an open field for description of pharmacophores against Leishmania parasites.
Incorporation of Laboratory Test Biomarkers Into Dual Antiplatelet Therapy Score Improves Prediction of Ischemic and Bleeding Events in Post-percutaneous Coronary Intervention Patients
Authors: Sun Chengming , Zhong Lin , Wu Yanqiu , Cao Chengfu , Guo Danjie , Liu Jie , Gong Lei , Zhang Shouxin , Sun Jun , Yu Yingqi , Tong Weiwei , Yang Jun
Published: May 2022

This study aimed to examine the performance of the dual antiplatelet therapy (DAPT) score in two retrospective cohorts of post-percutaneous coronary intervention (PCI) patients and to explore whether incorporating additional biomarkers could further improve the predictive power of the DAPT score. In a retrospective derivation cohort of 4,798 PCI patients, the validity of DAPT score for stratifying ischemic/bleeding risks was explored. Then, the association between the baseline status of 54 laboratory test biomarkers and ischemic/bleeding events was revealed while adjusting for the DAPT score. Combinations of individual laboratory test biomarkers that were significantly associated with ischemic/bleeding events were explored to identify the ones that improved discrimination of ischemic and bleeding events when incorporated into DAPT score. Finally, the impact of the combination of biomarkers with DAPT score was validated in an independent retrospective validation cohort of 1,916 PCI patients. Patients with a high DAPT score (DAPT score ≥ 2) had significantly higher risk of ischemic events and significantly lower risk of bleeding than patients with a low DAPT score (DAPT score < 2). Moreover, the addition of aspartate aminotransferase (AST) and red cell distribution width CV (RDW-CV) into the DAPT score further improved discrimination of ischemia and bleeding. Furthermore, the incremental predictive value of AST + RDW-CV maintained with measurements was updated at post-baseline time points. DAPT score successfully stratified the risks of ischemia/bleeding post PCI in the current cohorts. Incorporation of AST + RDW-CV into the DAPT score further improved prediction for both ischemic and bleeding events.
Risk for Cardiovascular Death Associated With Waist Circumference and Diabetes: A 9-Year Prospective Study in the Wan Shou Lu Cohort
Authors: Li Man , Zhu Ping , Wang Shu-xia
Published: May 2022

Background

It has been reported that obesity and diabetes are both risk factors for the development of cardiovascular diseases (CVDs). However, recent articles reported that compared with body mass index, waist circumference (WC) can better reflect obesity, more closely related to visceral fat tissue which is positively associated with an increased risk of cardiovascular death. Moreover, few studies have investigated the prognostic value of both WC and diabetes during a long-term follow-up. We aimed to investigate whether the higher level of WC measurements and diabetes were able to predict cardiovascular mortality in the general population.

Methods

In this prospective cohort study, a total of 1,521 consecutive subjects free of clinical CVD were included. The endpoint was cardiovascular death. The Kaplan–Meier method and Cox regression models were used to evaluate the cumulative risk of the outcome at different WC levels with or without diabetes.

Results

During a median follow-up of 9.2 years, 265 patients died due to cardiovascular conditions. Kaplan–Meier survival estimates indicated that the patients with higher levels of WC (WC > 94 cm) coexisted with diabetes had a significantly increased risk of cardiovascular death (log-rank p < 0.05). After adjustment for potential confounders, multiple COX regression models showed that the incidence of cardiovascular death was significantly higher when patients with high WC coexisted with diabetes mellitus (hazard ratio: 3.78; 95% CI: 3.35–3.98; p < 0.001).

Conclusion

Patients with high WC and diabetes represent a high-risk population for cardiovascular death. WC and diabetes may provide incremental prognostic value beyond traditional risks factors.
Realtime Remote Programming in Patients Carrying Cardiac Implantable Electronic Devices Requiring Emergent Reprogramming
Authors: Xiong Shiqiang , Li Jin , Tong Lin , Hou Jun , Yang Siqi , Qi Lingyao , Chen Xu , Luo Yan , Zhang Zhen , Liu Hanxiong , Cai Lin
Published: May 2022

To protect cardiac implantable electronic device (CIED) patients with arrhythmia or possible device malfunction, it is important for health care professionals to provide emergent device evaluation and reprogramming. This case series illustrated the clinical application of realtime remote programming in CIED patients requiring emergent in-person evaluation and reprogramming (ChiCTR2100046883 chictr.org). All remote sessions were performed safely and efficiently by remote electrophysiologists without being in the physical presence of a patient. The implementation of realtime remote programming not only largely reduces the response time to urgent events but also greatly helps to minimize personnel exposure to COVID-19 infection.
Social Contexts and Gender Disparities in Students’ Competence Beliefs: The Role of Gender-Stereotypical Beliefs and Achievement Patterns in the Classroom for Students’ Self-Concept in Gender-Stereotypical Subjects
Authors: Andersen Ida G. , Smith Emil
Published: May 2022

This study investigated the role of social contexts for gender disparities in education by examining the associations between gender-stereotypical beliefs (GSB) of students, peers, and teachers and gender achievement patterns in the classroom and students’ self-concept in language and math. We applied multilevel models with school fixed effects to a unique sample of combined survey and register data from Denmark to analyze detailed learning environments within schools and their correlations with gender differences in self-concept across subject domains. Results showed a gender gap in favor of boys in mathematics, net of academic achievement that were consistent across classrooms. In language, the influence of gender varied across classrooms. Furthermore, although GSB and gender achievement patterns did not alter the gender gap in either language or mathematics, we found that they moderated the relationship between gender and self-concept in heterogeneous ways across subjects. While teachers’ GSB increased the gender gap in language by decreasing boys’ self-concept, the students’ own GSB was more important for students’ self-concept in mathematics. Moreover, girls’ mathematics self-concept was lower in classrooms, in which, female peers had a relatively higher level of mathematics achievement compared to boys, suggesting that counter-stereotypical achievement patterns in the classroom do not increase students’ self-concept in subjects with strong gender stereotypes. On the contrary, girls are most likely to compare themselves to female peers, resulting in a negative association with self-evaluations. Our results highlighted the role played by social contexts in schools in the generation of gender differences in self-concept in traditionally stereotyped subject domains, but also showed important differences in how boys and girls were affected by their learning environments across different subject domains, suggesting there are different mechanisms at play.
Economic Burdens for Treatment of Patients With Type 2 Diabetes in North Thailand: A Hospital-Based Observational Study
Authors: Phrommintikul Arintaya , Dilokthornsakul Piyameth , Permsuwan Unchalee
Published: May 2022

Purpose

Diabetes and its complications pose an economic burden to healthcare systems, family, and society. Therefore, this study aimed to estimate the real-world financial burden of type 2 diabetes (T2D) treatment, complications, and cardiovascular death.

Materials and Methods

An electronic database of the largest university-affiliated hospital in the North of Thailand was retrieved for a 10-year period (2009-2019). We used the International Classification of Disease 10th Revision codes of diabetes and complications to obtain relevant patient records. All included records based on the inclusion and exclusion criteria were analyzed. Expenditures for diabetes treatment, complications, and cardiovascular death for two years were reported as mean, standard deviation, median, and interquartile range.

Results

Of a total of 9,161 patient records, the average age of patients was 57.8 ± 12.7 years. The average total outpatient cost was THB 22,874 ± 38,066 (US$ 759 ± 1,264) for the first year and THB 23,462 ± 34,441 (US$ 779 ± 1,143) for the second year. The average inpatient expenditure was THB 160,790 ± 411,607 (US$ 5,338 ± 13,666) for the first year and THB 181,804 ± 190,257 (US$ 6,036 ± 6,317) for the second year. Drug was the main component for outpatient expenditure while surgery was the main component for inpatient expenditure. Diabetes patients with complications incurred a greater cost of treatment than those without complications. Cardiovascular death led to about seven times higher cost of treatment than the average total cost of diabetes treatment. Heart failure complications (THB 846,345 ± 752,884 or US$ 28,099 ± 24,996) had the highest inpatient costs compared with other complications in the first year. Stroke complications (THB 71,927 ± 143,414 or US$ 2,388 ± 4,761) had the highest outpatient costs compared with other complications. In general, the first-year expenditure was higher than the second year for all complications.

Conclusions

Diabetes incurs a substantial financial burden resulting from its complications. Effective management of diabetes with a multi-sectoral effort from government, providers, patients, and private is required.
BRAFV600E Mutation-Responsive miRNA-222-3p Promotes Metastasis of Papillary Thyroid Cancer Cells via Snail-Induced EMT
Authors: Gao Yuan , Xiang Dapeng , Li Weijie , Zheng Xi , Wang Lin , Li Zhiyu , Chen Ting
Published: May 2022

BRAF mutation accounts for 50% of the PTC (papillary thyroid carcinoma) and is closely associated with high-risk clinicopathological characteristics. Increasing evidence implied that dysregulation of miRNA participated in carcinogenesis and progression of cancer. Clinical data showed the significant up-regulation of miR-222-3p in PTC; however, the role of miR-222-3p and possible relationship with BRAF mutation remained unclear. Here, we identified significant up-regulation of miR-222-3p in PTC tissues harboring BRAFV600E mutation compared with BRAF wild type (BRAFWT) from collected PTC clinical samples. External validation performed with The Cancer Genome Atlas (TCGA) databases was consistent with the above result. Exogenous expression of BRAFV600E oncoprotein increased the expression of miR-222-3p in B-CPAP and TPC-1 cells. The treatment of BRAFV600E and MEK inhibitor, PLX4720 and PD0325901, decreased the expression of miR-222-3p in B-CPAP but not in TPC-1. Inhibition of miR-222-3p significantly suppressed the migration of B-CPAP and induced a mesenchymal-epithelial transition (MET) phenotype via the Snail transcription factor. Immunohistochemistry (IHC) analysis demonstrated the up-regulation of Snail correlated with lymph node metastasis and BRAFV600E mutation in PTC. Besides, in situ hybridization (ISH) and IHC analysis of PTC clinical samples confirmed the correlation between the expression of miR-222-3p and Snail. These results showed miR-222-3p conduced more aggressive clinical manifestation of PTC by promoting Snail-induced EMT.
Analysis of Differential Diagnosis of Benign and Malignant Partially Cystic Thyroid Nodules Based on Ultrasound Characterization With a TIRADS Grade-4a or Higher Nodules
Authors: Wang Chen-Yi , Li Yang , Zhang Meng-Meng , Yu Zhi-Long , Wu Zi-Zhen , Li Chen , Zhang Dong-Chen , Ye Ying-Jiang , Wang Shan , Jiang Ke-Wei
Published: May 2022

Partially cystic thyroid nodules (PCTNs) are a kind of thyroid nodule with both solid and cystic components, and are usually misdiagnosed as benign nodules. The objective of this study was to determine the ultrasound (US) characterizations with a TIRADS Grade-4a or higher partially cystic thyroid nodules (PCTNs) which are associated with being malignant or benign. In this study, 133 PCTNs with a TIRADS Grade-4a or higher were enrolled; 83 were malignant and 50 were benign. TI-RADS classification can detect malignant PCTNs, and its sensitivity, specificity, positive predictive value, negative predictive value, and accuracy are 39.8%, 96.0%, 94.3%, 49.0%, and 60.9%, respectively. Univariate analyses revealed that nodule shape, margin, and structure were related to PCTNs’ being benign and malignant, among which nodules taller-than-wide, with an irregular shape, non-smooth margin, eccentric sharp angle, or edge sharp angle were significantly associated with malignancy while ovoid to round nodules, smooth margins, multiple separation, and eccentric obtuse angle structures were significantly associated with a benign nature. For the solid part of PCTNs, its free margin, echo, and calcification are related to benign and malignant PCTNs. Among them, the free margin of the solid part is non-smooth, hypoechoic, and microcalcification, which are related to malignant PCTNs, while the free margin of the solid part is smooth, isoechoic, macrocalcification, non-calcification and are related to benign PCTNs. Calcification of solid part and free margin are important factors for predicting malignant PCTNs. In addition, nodules’ composition, blood flow signal, and other factors had nothing to do with PCTNs’ being benign or malignant. In the multivariate Logistic regression analysis, solid part calcification (OR: 17.28; 95%CI: 5.14~58.08) and free margin (OR: 3.18; 95%CI: 1.01~10.00) were revealed to be the strongest independent predictors for malignancy (P<0.05). Our study indicated that understanding the ultrasound characteristics of malignant PCTNs, to avoid misdiagnosed PCTNs patients, is important to make a precise diagnosis and prognosis of PCTNs.
Association Between Fat Mass or Fat Fibrotic Gene Expression and Polyneuropathy in Subjects With Obesity: A Korean Metabolic Bariatric Surgery Cohort
Authors: Kim Kyuho , Oh Tae Jung , Park Young Suk , Chang Won , Cho Hyen Chung , Lee Jihye , Lee Yun Kyung , Choi Sung Hee , Jang Hak Chul
Published: May 2022

Aim

We aimed to investigate the association between obesity-related parameters and polyneuropathy (PN) and to evaluate inflammatory and fibrotic gene expression of fat as a potential mediator in subjects scheduled to undergo metabolic bariatric surgery (MBS).

Methods

This was a cross-sectional study of MBS cohort. Body composition and visceral fat area (VFA) were quantified by bioimpedance analysis and computed tomography scan. PN was defined by Michigan Neuropathy Screening Instrument–Physical Examination score was > 2. We measured mRNA expression level of FN1, TIMP1, CCL2, and CXCL8 in omental fat tissue.

Results

Of 189 subjects (mean age, 39.4 years; 69 [36.5%] male; mean body mass index, 38.5 kg/m2), prevalence of PN was 9.1% in subjects without diabetes (n = 110) and 20.3% in those with diabetes (n = 79). Nondiabetic subjects with PN had higher homeostatic model assessment-insulin resistance (6.8 ± 3.5 vs 4.5 ± 2.8, p = 0.041), and increased fat mass (58.5 ± 12.5 kg vs 50.5 ± 10.7 kg, p = 0.034), and VFA (309.4 ± 117.6 cm2vs 243.5 ± 94.2 cm2, p = 0.046) compared to those without PN. These obesity-related parameters were significantly associated with the presence of PN after adjusting for conventional risk factors of PN only in subjects without diabetes. In contrast, a fibrotic gene such as TIMP1 was independently associated with PN (adjusted odds ratio of 1.56; 95% confidence interval 1.06, 2.30) only in subjects with diabetes.

Conclusion

Increased adiposity was independently associated with PN in obese subjects without diabetes. In contrast, this association was not significant after adjusting conventional risk factors of PN in obese subjects with diabetes but increased fibrotic gene expression in fat was associated with PN in this group.
Contribution of Liver and Pancreatic Islet Crosstalk to β-Cell Function/Dysfunction in the Presence of Fatty Liver
Authors: López-Bermudo Lucía , Luque-Sierra Amparo , Maya-Miles Douglas , Gallego-Durán Rocío , Ampuero Javier , Romero-Gómez Manuel , Berná Genoveva , Martín Franz
Published: May 2022

Tissue-to-tissue crosstalk regulates organ function, according to growing data. This phenomenon is relevant for pancreatic β-cells and the liver, as both tissues are involved in glucose homeostasis and lipid metabolism. The ability to fine-tune regulation and adaptive responses is enabled through communication between pancreatic β-cells and the liver. However, the crosstalk between both tissues changes when metabolic dysregulation is present. Factors and cargo from extracellular vesicles (EVs) released by liver and pancreatic β-cells that reach the circulation form the words of this interaction. The molecules released by the liver are called hepatokines and are usually secreted in response to the metabolic state. When hepatokines reach the pancreatic islets several mechanisms are initiated for their protection or damage. In the case of the crosstalk between pancreatic β-cells and the liver, only one factor has been found to date. This protein, pancreatic derived factor (PANDER) has been proposed as a novel linker between insulin resistance (IR) and type 2 diabetes mellitus (T2D) and could be considered a biomarker for non-alcoholic fatty liver disease (NAFLD) and T2D. Furthermore, the cargo released by EVs, mainly miRNAs, plays a significant role in this crosstalk. A better knowledge of the crosstalk between liver and pancreatic β-cells is essential to understand both diseases and it could lead to better prevention and new therapeutic options.
Investigation of Fluid Excitation Forces Induced by Cylindrical and Conical Whirls in a Long Annular Seal Using Transient CFD Methods
Authors: Li Fengqin , Zhai Lulu , Cui Baoling , Zhu Zuchao , Guo Jia
Published: May 2022

Annular seals can induce fluid excitation forces that severely affect the stability of the pump rotor systems. In this paper, a transient computational fluid dynamic method with a mesh deformation technique is used to study the fluid excitation forces generated on the long annular seal caused by cylindrical whirls and conical whirls. The reliability of the method is verified by comparing the simulation results with the experimental results. The whirl motion, composed of cylindrical and conical whirls, is realized by the phase difference between the whirl movements at the annular seal inlet and outlet, and the effects of the phase difference and inlet preswirl on the fluid excitation forces are studied numerically. The results show that the fluid excitation forces significantly depend on the phase difference of the rotor whirl movement, and tangential force acts as a stabilizing force only when the phase difference approaches 45°. The tangential forces of the cylindrical whirl, the conical whirl and the form of cylindrical and conical combined whirl are sensitive to inlet preswirl, while the effect of inlet preswirl on the radial forces becomes obvious only when the rotor whirls in the form of combined whirls.
The Symmetric and Asymmetric Effect of Defense Expenditures, Financial Liberalization, Health Expenditures on Sustainable Development
Authors: Meiling Li , Taspinar Nigar , Yahya Farzan , Hussain Muhammad , Waqas Muhammad
Published: May 2022

This study examines the short run, long run, and causal relationships among financial liberalization, healthcare expenditures, and defense expenditures on sustainable development in Pakistan covering the period from 1971 to 2017. The paper also explored the asymmetric relationships among the target variables. To explore these relationships, ARDL and NARDL Models are utilized. Additionally, advanced econometric techniques such as Maki cointegration and quasi-GLS unit root are used to take multiple structural breaks into account. Maki cointegration results show a stable long run relationship between the underlying variables. The findings of ARDL suggest a positive effect of financial liberalization and health expenditures while the negative effect of military expenditures on sustainable development. NARDL estimates suggest strong asymmetry as sustainability responds to positive (negative) shocks in militarization, health expenditures, and financial liberalization differently. The Toda-Yamamoto causality test shows that any policy to target health expenditures and financial liberalization significantly alters sustainable development and vice versa. For robustness checks, FMOLS and alternative proxy of sustainable development are used. The key findings posited the need to shift military expenditures to health expenditures and financial markets to achieve sustainable development goals in Pakistan.
On-Campus Field Experiences Help Students to Learn and Enjoy Water Science During the COVID-19 Pandemic
Authors: Saup C. , Lamantia K. , Chen Z. , Bell B. , Schulze J. , Alsdorf D. , Sawyer A.H.
Published: May 2022

Online modes of teaching and learning have gained increased attention following the COVID-19 pandemic, resulting in education delivery trends likely to continue for the foreseeable future. It is therefore critical to understand the implications for student learning outcomes and their interest in or affinity towards the subject, particularly in water science classes, where educators have traditionally employed hands-on outdoor activities that are difficult to replicate online. In this study, we share our experiences adapting a field-based laboratory activity on groundwater to accommodate more than 700 students in our largest-enrollment general education course during the pandemic. As part of our adaptation strategy, we offered two versions of the same exercise, one in-person at the Mirror Lake Water Science Learning Laboratory, located on Ohio State University’s main campus, and one online. Although outdoor lab facilities have been used by universities since at least the 1970s, this research is novel in that 1) it considers not only student achievement but also affinity for the subject, 2) it is the first of its kind on The Ohio State University’s main campus, and 3) it was conducted during the COVID-19 pandemic, at a time when most university classes were unable to take traditional field trips. We used laboratory grades and a survey to assess differences in student learning and affinity outcomes for in-person and online exercises. Students who completed the in-person exercise earned better scores than their online peers. For example, in Fall 2021, the median lab score for the in-person group was 97.8%, compared to 91.7% for the online group. The in-person group also reported a significant (p < 0.05) increase in how much they enjoyed learning about water, while online students reported a significant decrease. Online students also reported a significant decrease in how likely they would be to take another class in water or earth sciences. It is unclear whether the in-person exercise had better learning and affinity outcomes because of the hands-on, outdoor qualities of the lab or because the format allowed greater interaction among peers and teaching instructors (TAs). To mitigate disparities in student learning outcomes between the online and in-person course delivery, instructors will implement future changes to the online version of the lab to enhance interactions among students and TAs.
Revisiting the EKC Hypothesis With Export Diversification and Ecological Footprint Pressure Index for India: A RALS-Fourier Cointegration Test
Authors: Pata UK , Shahzad Farrukh , Fareed Zeeshan , Rehman MA
Published: May 2022

This study assesses India’s environmental problems by focusing on the ecological footprint pressure index (EFPI), which provides a simultaneous analysis of biocapacity and ecological footprint. In particular, the study examines the impact of export diversification, economic growth, and renewable energy on EFPI under the environmental Kuznets curve (EKC) hypothesis for India. To analyze the long-run relationships, the Residual Augmented Ordinary Least Squares (RALS)-Fourier cointegration test is applied to annual data from 1965 to 2014. Contrary to the EKC hypothesis, the results show a U-shaped relationship between income and EFPI. The absence of the EKC hypothesis implies that economic development does not help to ensure environmental quality; hence, other measures are required to minimize irreversible environmental problems. In this regard, the results of the study suggest that renewable energy consumption and export diversification reduce EFPI. Based on the overall findings of the study, it is recommended that the Indian government improve environmental values by changing the export structure and energy mix.
Fiscal Decentralization, Government Behavior, and Environmental Pollution: Evidence From China
Authors: Cai Hongying , Tong Zefeng , Xu Shulin , Chen Shuoqi , Zhu Peng , Liu Wenjie
Published: May 2022

Environmental protection is a basic public service that the government must guarantee and is closely related to public health. An important driver of environmental pollution in China is the local government’s pursuit of a rapid economic development while ignoring environmental protection under the Chinese-style fiscal decentralization system. On the basis of the principal–agent theory between the central and local governments, this study analyzes the environmental deterioration caused by the distortion of local government behavior under fiscal decentralization. In addition, using China’s prefecture-level city data from 2014 to 2018, this study empirically estimates the impact of fiscal decentralization on environmental pollution. SO2 emissions and PM2.5 concentrations are used to measure the degree of environmental pollution. Results show that Chinese-style fiscal decentralization exacerbates environmental pollution and that the impact of fiscal decentralization on environmental pollution differs in regions with varying levels of economic development and cultural penetration. Moreover, fiscal decentralization does not significantly impact environmental pollution in eastern China and in those areas influenced by Confucian culture yet aggravates the environmental pollution in central and western China and in those areas that are not affected by Confucian culture. These results offer important policy implications. Clearly dividing the power and financial power between the central and local governments, establishing an environmental governance system compatible with economic incentives, and building an environmental public finance system can alleviate the impact of Chinese-style fiscal decentralization on environmental pollution.
The Deubiquitinating Enzyme MrUbp14 Is Involved in Conidiation, Stress Response, and Pathogenicity in Metarhizium robertsii
Authors: Wang Zhangxun , Chen Hua , Li Hao , Chen Hanyuan , Huang Bo
Published: May 2022

Protein ubiquitination, which is involved in various biological processes in eukaryotic cells, is a reversible modification of proteins. Deubiquitinases can maintain ubiquitin homeostasis by removing ubiquitin or modulating protein degradation via the ubiquitin-proteasome system (UPS). Metarhizium robertsii, an entomopathogenic fungus, has become a model fungus for investigating the interactions between insects and fungal pathogens. To explore the possible effects of the deubiquitination process on the development, stress response, and virulence of M. robertsii, disruption of MrUbp14 (an ortholog of the yeast ubiquitin-specific protease gene, Ubp14) was performed. The results of this study showed that the deletion of MrUbp14 led to accelerated conidial germination, reduced conidial yields, and decreased expression levels of some genes involved in conidiation. Furthermore, the MrUbp14 mutant (ΔMrUbp14) exhibited decreased tolerance to cell wall-damaging stressors (Congo red and SDS) and heat stress. Importantly, the results of the bioassay demonstrated that the fungal virulence of the ΔMrUbp14 strain was largely reduced in cuticle infection, but not in direct injection, which was accompanied by a significant decline in appressorium formation and cuticle penetration. Moreover, our results demonstrated that the disruption of MrUbp14 resulted in significantly increased ubiquitination levels of total protein, suggesting that MrUbp14 acts as a deubiquitinating enzyme in M. robertsii. In summary, our phenotypic changes in the gene disruption mutants suggest that MrUbp14 is important for conidiation, stress response, and fungal virulence in M. robertsii.
Primary Cell Lines From Feathers and Blood of Free-Living Tawny Owls (Strix aluco): A New In Vitro Tool for Non-Lethal Toxicological Studies
Authors: Kroglund Ingvild Buran , Eide Sara Kristiane Kjærgård , Østnes Jan Eivind , Kroglund Rolf Terje , Frisli Jan-Erik , Waugh Courtney Alice
Published: May 2022

The validation of the use of primary cell lines from non-lethal matrixes of feathers and blood of nestlings of a wild bird species, the tawny owl (Strix aluco) is described. Tawny Owl Feather Fibroblast (TOFF) cells and peripheral blood mononuclear cells (PBMCs) were isolated and cultured from the pulp of the secondary wing feathers and whole blood respectively from free-living tawny owl nestlings. Cell growth was registered up until 48 h for both the PBMC cells and the TOFFs. The validation of these primary cell lines in free-living birds has the potential to advance the assessment of immunotoxicological effects in wildlife via non-lethal manner. They provide a key tool with which to study cell toxicity and responses to environmental stressors on a cellular level in wild bird species of interest.
Identifying Candidate Genes for Short Gestation Length Trait in Chinese Qingping Pigs by Whole-Genome Resequencing and RNA Sequencing
Authors: Liu Zezhang , Yang Jun , Li Hong , Zhong Zhuxia , Huang Jian , Fu Jie , Zhao Hucheng , Liu Xiaolei , Jiang Siwen
Published: May 2022

Gestation length is a complex polygenic trait that affects pig fetal development. The Qingping (QP) pig, a Chinese native black pig breed, is characterized by short gestation length. However, the genetic architecture of short gestation length is still not clear. The present study aimed to explore the genetic architecture of short gestation length in QP pigs. In this study, selective sweep analyses were performed to detect selective sweep signatures for short gestation length traits between 100 QP pigs and 219 pigs from 15 other breeds. In addition, differentially expressed genes for the short gestation length between QP pigs and Large White pigs were detected by RNA sequencing. Comparing candidate genes from these methods with known genes for preterm birth in the database, we obtained 111 candidate genes that were known preterm birth genes. Prioritizing other candidate genes, 839 novel prioritized candidate genes were found to have significant functional similarity to preterm birth genes. In particular, we highlighted EGFR, which was the most prioritized novel candidate relative to preterm birth genes. Experimental validations in placental and porcine trophectoderm cells suggest that EGFR is highly expressed in the QP pigs with short gestation length and could regulate the NF-κΒ pathway and downstream expression of PTGS2. These findings comprehensively identified candidate genes for short gestation length trait at the genomic and transcriptomic levels. These candidate genes provide an important new resource for further investigation and genetic improvement of gestation length.
Novel Missense and Splice Site Mutations in USH2A, CDH23, PCDH15, and ADGRV1 Are Associated With Usher Syndrome in Lebanon
Authors: Jaffal Lama , Akhdar Hanane , Joumaa Hawraa , Ibrahim Mariam , Chhouri Zahraa , Assi Alexandre , Helou Charles , Lee Hane , Seo Go Hun , Joumaa Wissam H. , El Shamieh Said
Published: May 2022

The purpose of this study was to expand the mutation spectrum by searching the causative mutations in nine Lebanese families with Usher syndrome (USH) using whole-exome sequencing. The pathogenicity of candidate mutations was first evaluated according to their frequency, conservation, and in silico prediction tools. Then, it was confirmed via Sanger sequencing, followed by segregation analysis. Finally, a meta-analysis was conducted to calculate the prevalence of USH genes in the Lebanese population. Three missense mutations, two splice site mutations, and one insertion/deletion were detected in eight of the families. Four of these variants were novel: c.5535C > A; p.(Asn1845Lys) in exon 41 of CDH23, c.7130G > A; p.(Arg2377Gln) in exon 32 of ADGRV1, c.11390-1G > A in USH2A, and c.3999–6A > G in PCDH15. All the identified mutations were shown to be likely disease-causing through our bioinformatics analysis and co-segregated with the USH phenotype. The mutations were classified according to the ACMG standards. Finally, our meta-analysis showed that the mutations in ADGRV1, USH2A, and CLRN1 are the most prevalent and responsible for approximately 75% of USH cases in Lebanon. Of note, the frequency USH type 3 showed a relatively high incidence (23%) compared to the worldwide prevalence, which is around 2–4%. In conclusion, our study has broadened the mutational spectrum of USH and showed a high heterogeneity of this disease in the Lebanese population.
Spectrum of Mutations in NDP Resulting in Ocular Disease; a Systematic Review
Authors: Wawrzynski James , Patel Aara , Badran Abdul , Dowell Isaac , Henderson Robert , Sowden Jane C.
Published: May 2022

Aims and Rationale: The inner retina is supplied by three intraretinal capillary plexi whereas the outer retina is supplied by the choroidal circulation: NDP is essential for normal intraretinal vascularisation. Pathogenic variants in NDP (Xp11.3) may result in either a severe retinal phenotype associated with hearing loss (Norrie Disease) or a moderate retinal phenotype (Familial Exudative Vitreoretinopathy, FEVR). However, little is known about whether the nature or location of the NDP variant is predictive of severity. In this systematic review we summarise all reported NDP variants and draw conclusions about whether the nature of the NDP variant is predictive of the severity of the resulting ocular pathology and associated hearing loss and intellectual disability.

Findings: 201 different variants in the NDP gene have been reported as disease-causing. The pathological phenotype that may result from a disease-causing NDP variant is quite diverse but generally comprises a consistent cluster of features (retinal hypovascularisation, exudation, persistent foetal vasculature, tractional/exudative retinal detachment, intellectual disability and hearing loss) that vary predictably with severity. Previous reviews have found no clear pattern in the nature of NDP mutations that cause either FEVR or Norrie disease, with the exception that mutations affecting cysteine residues have been associated with Norrie Disease and that visual loss amongst patients with Norrie disease tends to be more severe if the NDP mutation results in an early termination of translation as opposed to a missense related amino acid change. A key limitation of previous reviews has been variability in the case definition of Norrie disease and FEVR amongst authors. We thus reclassified patients into two groups based only on the severity of their retinal disease. Of the reported pathogenic variants that have been described in more than one patient, we found that any given variant caused an equivalent severity of retinopathy each time it was reported with very few exceptions. We therefore conclude that specific NDP mutations generally result in a consistent retinal phenotype each time they arise. Reports by different authors of the same variant causing either FEVR or Norrie disease conflict primarily due to variability in the authors’ respective case definitions rather than true differences in disease severity.
Determining the Area of Ancestral Origin for Individuals From North Eurasia Based on 5,229 SNP Markers
Authors: Gorin Igor , Balanovsky Oleg , Kozlov Oleg , Koshel Sergey , Kostryukova Elena , Zhabagin Maxat , Agdzhoyan Anastasiya , Pylev Vladimir , Balanovska Elena
Published: May 2022

Currently available genetic tools effectively distinguish between different continental origins. However, North Eurasia, which constitutes one-third of the world’s largest continent, remains severely underrepresented. The dataset used in this study represents 266 populations from 12 North Eurasian countries, including most of the ethnic diversity across Russia’s vast territory. A total of 1,883 samples were genotyped using the Illumina Infinium Omni5Exome-4 v1.3 BeadChip. Three principal components were computed for the entire dataset using three iterations for outlier removal. It allowed the merging of 266 populations into larger groups while maintaining intragroup homogeneity, so 29 ethnic geographic groups were formed that were genetically distinguishable enough to trace individual ancestry. Several feature selection methods, including the random forest algorithm, were tested to estimate the number of genetic markers needed to differentiate between the groups; 5,229 ancestry-informative SNPs were selected. We tested various classifiers supporting multiple classes and output values for each class that could be interpreted as probabilities. The logistic regression was chosen as the best mathematical model for predicting ancestral populations. The machine learning algorithm for inferring an ancestral ethnic geographic group was implemented in the original software “Homeland” fitted with the interface module, the prediction module, and the cartographic module. Examples of geographic maps showing the likelihood of geographic ancestry for individuals from different regions of North Eurasia are provided. Validating methods show that the highest number of ethnic geographic group predictions with almost absolute accuracy and sensitivity was observed for South and Central Siberia, Far East, and Kamchatka. The total accuracy of prediction of one of 29 ethnic geographic groups reached 71%. The proposed method can be employed to predict ancestries from the populations of Russia and its neighbor states. It can be used for the needs of forensic science and genetic genealogy.
Uncovering BTB and CNC Homology1 (BACH1) as a Novel Cancer Therapeutic Target
Authors: Liu Zheming , Wang Jing , Chen Huiyong , Wu Zankai , Liao Fuben , Wang Sheng , Zhu Ting
Published: May 2022

BTB and CNC homology1 (BACH1), working as a transcriptional factor, is demonstrated to function on the regulation of epigenetic modifications by complex regulatory networks. Although BACH1 is reported as an oncogene, the overall analysis of its role remains lacking. In this study, we uncovered the capacity of BACH1 as a new pan-cancer therapeutic target. We found that BACH1 is highly expressed in abundant cancers and correlated with the poor prognosis of most cancers. The mutation sites of BACH1 varied in different cancer types and correlated to patients’ prognoses. The tumor mutation burden (TMB) in four cancer species and up to six tumor infiltrated immune cells had a significant relevance with BACH1. The enrichment analysis showed that the BACH1-associated genes were significantly enriched in the pathways of PD-1/L1 expression, ubiquitin-mediated proteolysis, T cell receptor, Th17 cell differentiation. We then demonstrated that BACH1 is positively correlated with the expression of many candidate genes, incluing SRPK2, GCLM, SLC40A1, and HK2 but negatively correlated with the expression of KEAP1 and GAPDH. Overall, our data shed light on BACH1’s effect on latent utility in cancer targeting therapy.