bluuebunny/crossref_metadata_2025 huggingface dataset:
Prenatal Genetic Diagnosis in Three Fetuses With Left Heart Hypoplasia (LHH) From Three Unrelated Families
Authors: Luo Sukun , Chen Luyi , Wei Weizhong , Tan Li , Zhang Meng , Duan Zhengrong , Cao Jiangxia , Zhou Yan , Zhou Aifen , He Xuelian
Published: April 2021

Background: Congenital heart defects (CHDs) are the most common birth defects, and left heart hypoplasia (LHH) is a severe form of CHD and responsible for more than 20% cardiac deaths during the first week of life, however, its genetic causes remain largely elusive.

Methods: Three families with fetal LHH were recruited. Genomic DNA from amniotic fluid or peripheral blood, and trio whole exome sequencing (trio-WES) and copy number variation sequencing (CNV-seq) were performed.

Results: All the three couples had no family history, and mid-gestation ultrasound revealed LHH and other variable cardiovascular defects in the fetuses. Trio-WES revealed de novo pathogenic variations in KMT2D (p.Gly3465Aspfs*37) (NM_003482) and WDFY3 (p.Ser117Xfs*) (NM_014991), and CNV-seq identified a deletion of 150 kb encompassing NOTCH1. KMT2D and NOTCH1 previously have been reported to be associated with CHDs, however, WDFY3 is reported for the first time to be possibly related to CHD in human.

Conclusion: Our study suggested that genetic component is an important risk factor for the development of LHH, and next generation sequencing is a powerful tool for genetic diagnosis in fetuses with CHDs and genetic counseling, however, more studies and data are need to establish the correlation of fetal phenotypes and genotypes.
How Personality Traits Are Related to the Attitudes Toward Forced Remote Learning During COVID-19: Predictive Analysis Using Generalized Additive Modeling
Authors: Dikaya Liudmila A. , Avanesian Garen , Dikiy Igor S. , Kirik Vladimir A. , Egorova Valeria A.
Published: April 2021

Background

During the COVID-19 pandemic, universities all over the world have had to reorganize their work for remote education to ensure continuity of learning. This situation has forced both teachers and students into an atypical, very specific situation, in which they need to cope with a number of psychological factors. Meanwhile, there is a research gap in academic knowledge of the social and psychological factors that determine attitudes toward forced remote learning.

Objective

To analyze the psychological traits of students in relation to their attitudes toward forced remote learning.

Methods

The study assessed self-regulation and communication skills, as well thinking and learning styles of 280 students in the situation of forced remote learning. We used the methods of surveying and psychological testing for data collection. The data were analyzed in R statistical software using the regression modeling methods.

Results

We found that the number of students with positive (46%) and negative (54%) attitudes toward forced remote learning during the COVID-19 pandemic was approximately equal. Using regression analysis, we identified several statistically significant associations between the interpersonal communicative skills of students (self-regulation, shyness, alienation, manipulative and cooperative communication styles) and their thinking styles (right-hemispheric and integrated), on the one hand, and their attitude to remote learning, on the other. It was also illustrated that depending on the attitude to the forced remote learning, students differ by the percentage of assimilated learning materials while studying under the lockdown.

Conclusion

The results identify that success of remote learning in many ways depends on the extent to which it accommodates psychological traits of students who are forced to acquire new knowledge without traditional classroom instruction.
Mitigation of Acute Ammonia Emissions With Biochar During Swine Manure Agitation Before Pump-Out: Proof-of-the-Concept
Authors: Chen Baitong , Koziel Jacek A. , Białowiec Andrzej , Lee Myeongseong , Ma Hantian , O’Brien Samuel , Li Peiyang , Meiirkhanuly Zhanibek , Brown Robert C.
Published: April 2021

Ammonia (NH3) emissions from animal agriculture can cause eutrophication of water ecosystems and are precursors to secondary particulate matter (PM2.5). NH3 emissions from stored swine manure represent nutrient loss affecting the fertilizing value of manure. The short-term emission bursts occur when farmers agitate manure before emptying storage and fertilizing fields. There is no proven technology to mitigate gaseous emissions during agitation, while the hazards of acute releases (e.g., H2S) are well-known. Biochar mitigates NH3 emissions from manure over the long-term. The objective of this research was to evaluate the mitigation of acute NH3 emissions during/after agitation. Two biochars, highly alkaline and porous (HAP from corn stover) and red oak (RO), were tested. The 6 and 12 mm-thick layers of biochar powder were surficial applied followed by 3 min agitation. NH3 concentrations were measured before/during/after agitation. Mitigation was assessed by comparing: (i) the maximum (peak) flux, (ii) total emission (from agitation start till NH3 concentration returned to the before-agitation), and (iii) the total emissions during agitation. The 12 mm HAP significantly (p < 0.05) reduced (i–iii) by 63, 70, and 85%, respectively. The 6 mm HAP significantly reduced (i–iii) by 76, 75, and 78%, respectively. The 12 mm RO significantly reduced (i–iii) by 9, 53, and 57%, respectively. The 6 mm RO significantly reduced (i–iii) by 61, 86, and 63%, respectively. The NH3 emission kinetics model confirmed that a 6 mm dose was just as effective as the larger dose. More research is needed to optimize and scale-up mitigating emissions and retention of nutrients in manure with biochar.
Bark-Water Interactions Across Ecosystem States and Fluxes
Authors: Van Stan John T. , Dymond Salli F. , Klamerus-Iwan Anna
Published: April 2021

To date, the perspective of forest ecohydrologists has heavily focused on leaf-water interactions – leaving the ecohydrological roles of bark under-studied, oversimplified, or omitted from the forest water cycle. Of course, the lack of study, oversimplification, or omission of processes is not inherently problematic to advancing ecohydrological theory or operational practice. Thus, this perspective outlines the relevance of bark-water interactions to advancing ecohydrological theory and practice: (i) across scales (by briefly examining the geography of bark); (ii) across ecosystem compartments (i.e., living and dead bark on canopies, stems, and in litter layers); and, thereby, (iii) across all major hydrologic states and fluxes in forests (providing estimates and contexts where available in the scant literature). The relevance of bark-water interactions to biogeochemical aspects of forest ecosystems is also highlighted, like canopy-soil nutrient exchanges and soil properties. We conclude that a broad ecohydrological perspective of bark-water interactions is currently merited.
TOX Acts as a Tumor Suppressor by Inhibiting mTOR Signaling in Colorectal Cancer
Authors: Yang Mengdi , Huang Qianru , Li Changcan , Jiang Zhiyuan , Sun Jing , Wang Zhiyu , Liang Rui , Li Dan , Li Bin , Zhao Hui
Published: April 2021

The treatment and prognosis of advanced colorectal cancer (CRC) remain a challenging clinical research focus. Here, we describe a new CRC tumor suppressor and potential therapeutic target: thymocyte selection associated high mobility group box (TOX) protein. The expression of TOX was lower in CRC than para-CRC. With the increase of tumor stage, TOX expression decreased, indicating the presence of TOX relates to better overall survival (OS). TOX suppressed the mechanistic target of rapamycin kinase (mTOR) signaling to inhibit cell proliferation, migration, invasion, and change the epithelial-mesenchymal transition (EMT) process. In addition, TOX promoted apoptosis. As tumor mutation burden and tumor microenvironment play vital roles in the occurrence and development of tumors, we analyzed the TOX expression in the immune microenvironment of CRC. The high TOX expression was negatively correlated with TumorPurity. Moreover, it was positively related to ImmuneScore, StromalScore, microsatellite instability (MSI) status, and Consensus Molecular Subtypes (CMS) 3 typing. Based on gene set enrichment analysis (GSEA), the reduced expression of TOX activated mTOR. We found rapamycin, a mTOR inhibitor, partly inhibited cell proliferation, invasion, and migration in shTOX HCT116 cells. Lastly, TOX suppressed tumorigenesis and lung metastasis of CRC in vivo. Rapamycin alone or combined with PD1 inhibitor is more effective than PD1 inhibitor alone in a tumor model. Taken together, these findings highlight the tumor-suppressive role of TOX in CRC, especially in MSI CRC, and provide valuable information that rapamycin alone or combined with PD1 inhibitor has therapeutic potential in CRC.
An Improved Equivalent Circuit Simulation of High Frequency Ultrasound Transducer
Authors: Shen Zhitian , Xu Jie , Li Zhangjian , Chen Youwei , Cui Yaoyao , Jian Xiaohua
Published: April 2021

The equivalent circuit simulation plays an important role in the design of ultrasound transducer. However, the existing methods are difficult to achieve the effect of matching and backing layer, and not able to accurately simulate the transducer with cable. Especially in the application of high frequency ultrasound, the long cable has a great influence on the performance of the transducer. To overcome these limitations, this paper proposed an improved equivalent circuit method, which combined Leach model and transmission line model. It can realize the complete simulation of ultrasound transducer with a long cable, matching layer, and backing layer in PSPICE circuit simulation software when the parameters were measured. Its principles were briefly introduced, and ultrasound transducers with different frequencies (12 and 20 MHz), different matching layers, and different cable lengths (0.5–2.5 m) were designed and fabricated to verify the effectiveness of the method, which is also compared with the traditional KLM method using PiezoCAD. The experiment results showed that the long cable, matching layer, and backing layer have a significant impact on the performance of high frequency ultrasound transducers, and this proposed method has good agreement with these results. Moreover, for the simulation of the complete transducer, the effect of this method is better than KLM model. Besides, this method does not need to know the specific equivalent circuit of matching, backing layer, or cable wire, it can accurately predict the impedance and phase of the transducer through the material parameters, which is very helpful for the material selection and optimization of subsequent transducer design and fabrication. The study indicates that this improved equivalent circuit method is suitable to be applied in the general circuit simulation software and provides strong support for the high frequency transducer and system design.
Natural History of Clinical, Laboratory, and Echocardiographic Parameters of a Primary Hyperoxaluria Cohort on Long Term Hemodialysis
Authors: Sas David J. , Enders Felicity T. , Gunderson Tina M. , Mehta Ramila A. , Olson Julie B. , Seide Barbara M. , Banks Carly J. , Dehmel Bastian , Pellikka Patricia A. , Lieske John C. , Milliner Dawn S.
Published: April 2021

Background: Primary hyperoxaluria type 1 (PH1) is a rare monogenic disorder characterized by excessive hepatic production of oxalate leading to recurrent nephrolithiasis, nephrocalcinosis, and progressive kidney damage, often requiring renal replacement therapy (RRT). Though systemic oxalate deposition is well-known, the natural history of PH1 during RRT has not been systematically described. In this study, we describe the clinical, laboratory, and echocardiographic features of a cohort of PH1 patients on RRT.

Methods: Patients with PH1 enrolled in the Rare Kidney Stone Consortium PH Registry who progressed to require RRT, had ≥2 plasma oxalate (pOx) measurements 3–36 months after start of RRT, and at least one pair of pOx measurements between 6 and 18 months apart were retrospectively analyzed. Clinical, echocardiographic, and laboratory results were obtained from the Registry.

Results: The 17 PH1 patients in our cohort had a mean total HD hours/week of 17.4 (SD 7.9; range 7.5–36) and a range of age of RRT start of 0.2–75.9 years. The average change in plasma oxalate (pOx) over time on RRT was −0.74 [−2.9, 1.4] μmol/L/month with the mean pOx never declining below 50 μmol/L. Over time on RRT, oxalosis progressively developed in multiple organ systems. Echocardiography performed on 13 subjects showed worsening of left ventricular global longitudinal strain correlated with pOx (p < 0.05).

Conclusions: Even when a cohort of PH1 patients were treated with intensified RRT, their predialysis pOx remained above target and they developed increasing evidence of oxalosis. Echocardiographic data suggest that cardiac dysfunction could be related to elevated pOx and may worsen over time.
Identification and Expression of Secreted In Xylem Pathogenicity Genes in Fusarium oxysporum f. sp. pisi
Authors: Jenkins Sascha , Taylor Andrew , Jackson Alison C. , Armitage Andrew D. , Bates Helen J. , Mead Andrew , Harrison Richard J. , Clarkson John P.
Published: April 2021

Fusarium oxysporum is a soilborne fungal plant pathogen responsible for causing disease in many economically important crops with “special forms” (formae speciales) adapted to infect specific plant hosts. F. oxysporum f. sp. pisi (FOP) is the causal agent of Fusarium wilt disease of pea. It has been reported in every country where peas are grown commercially. Disease is generally controlled using resistant cultivars possessing single major gene resistance and therefore there is a constant risk of breakdown. The main aim of this work was to characterise F. oxysporum isolates collected from diseased peas in the United Kingdom as well as FOP isolates obtained from other researchers representing different races through sequencing of a housekeeping gene and the presence of Secreted In Xylem (SIX) genes, which have previously been associated with pathogenicity in other F. oxysporum f. spp. F. oxysporum isolates from diseased United Kingdom pea plants possessed none or just one or two known SIX genes with no consistent pattern of presence/absence, leading to the conclusion that they were foot-rot causing isolates rather than FOP. In contrast, FOP isolates had different complements of SIX genes with all those identified as race 1 containing SIX1, SIX6, SIX7, SIX9, SIX10, SIX11, SIX12, and SIX14. FOP isolates that were identified as belonging to race 2 through testing on differential pea cultivars, contained either SIX1, SIX6, SIX9, SIX13, SIX14 or SIX1, SIX6, SIX13. Significant upregulation of SIX genes was also observed in planta over the early stages of infection by different FOP races in pea roots. Race specific SIX gene profiling may therefore provide potential targets for molecular identification of FOP races but further research is needed to determine whether variation in complement of SIX genes in FOP race 2 isolates results in differences in virulence across a broader set of pea differential cultivars.
Improvement of Free Fatty Acid Secretory Productivity in Aspergillus oryzae by Comprehensive Analysis on Time-Series Gene Expression
Authors: Wong Pui Shan , Tamano Koichi , Aburatani Sachiyo
Published: April 2021

Aspergillus oryzae is a filamentous fungus that has historically been utilized in the fermentation of food products. In recent times, it has also been introduced as a component in the industrial biosynthesis of consumable compounds, including free fatty acids (FFAs), which are valuable and versatile products that can be utilized as feedstocks in the production of other commodities, such as pharmaceuticals and dietary supplements. To improve the FFA secretory productivity of A. oryzae in the presence of Triton X-100, we analyzed the gene expression of a wild-type control strain and a disruptant strain of an acyl-CoA synthetase gene, faaA, in a time-series experiment. We employed a comprehensive analysis strategy using the baySeq, DESeq2, and edgeR algorithms to clarify the vital pathways for FFA secretory productivity and select genes for gene modification. We found that the transport and metabolism of inorganic ions are crucial in the initial stages of FFA production and revealed 16 candidate genes to be modified in conjunction with the faaA disruption. These genes were verified through the construction of overexpression strains, and showed that the manipulation of reactions closer to the FFA biosynthesis step led to a higher increase in FFA secretory productivity. This resulted in the most successful overexpression strains to have an FFA secretory productivity more than two folds higher than that of the original faaA disruptant. Our study provides guidance for further gene modification for FFA biosynthesis in A. oryzae and for enhancing the productivity of other metabolites in other microorganisms through metabolic engineering.
Electron & Biomass Dynamics of Cyanothece Under Interacting Nitrogen & Carbon Limitations
Authors: Rabouille Sophie , Campbell Douglas A. , Masuda Takako , Zavřel Tomáš , Bernát Gábor , Polerecky Lubos , Halsey Kimberly , Eichner Meri , Kotabová Eva , Stephan Susanne , Lukeš Martin , Claquin Pascal , Bonomi-Barufi José , Lombardi Ana Teresa , Červený Jan , Suggett David J. , Giordano Mario , Kromkamp Jacco C. , Prášil Ondřej
Published: April 2021

Marine diazotrophs are a diverse group with key roles in biogeochemical fluxes linked to primary productivity. The unicellular, diazotrophic cyanobacterium Cyanothece is widely found in coastal, subtropical oceans. We analyze the consequences of diazotrophy on growth efficiency, compared to NO3–-supported growth in Cyanothece, to understand how cells cope with N2-fixation when they also have to face carbon limitation, which may transiently affect populations in coastal environments or during blooms of phytoplankton communities. When grown in obligate diazotrophy, cells face the double burden of a more ATP-demanding N-acquisition mode and additional metabolic losses imposed by the transient storage of reducing potential as carbohydrate, compared to a hypothetical N2 assimilation directly driven by photosynthetic electron transport. Further, this energetic burden imposed by N2-fixation could not be alleviated, despite the high irradiance level within the cultures, because photosynthesis was limited by the availability of dissolved inorganic carbon (DIC), and possibly by a constrained capacity for carbon storage. DIC limitation exacerbates the costs on growth imposed by nitrogen fixation. Therefore, the competitive efficiency of diazotrophs could be hindered in areas with insufficient renewal of dissolved gases and/or with intense phytoplankton biomass that both decrease available light energy and draw the DIC level down.
Anti-biofilm Potential of Elletaria cardamomum Essential Oil Against Escherichia coli O157:H7 and Salmonella Typhimurium JSG 1748
Authors: Abdullah  , Asghar Ali , Algburi Ammar , Huang Qingrong , Ahmad Talha , Zhong Hao , Javed Hafiz U. , Ermakov Alexey M. , Chikindas Michael L.
Published: April 2021

Foodborne pathogens, microbial recurrent infections, and antibiotic resistance have driven researchers to explore natural compounds as safe alternative antimicrobials. In this study, the chemical profile, antimicrobial, and mutagenic activities of the Elletaria cardamomum essential oil were investigated. GC-MS analysis identified the major bioactive components as α-terpinyl acetate, 1,8-cineole, linalool acetate, and sabinene, at concentrations of 34.95, 25.30, 8.13, and 5.48% respectively, of the essential oil’s content. Regarding antimicrobial activity, the minimum inhibitory concentration of green cardamom essential oil was 1% against Escherichia coli O157:H7 and Pseudomonas aeruginosa ATCC 14213. Green cardamom essential oil, when used at concentrations of 0.015, 0.031, 0.062, and 0.125% (v/v) prevented biofilm formation of Escherichia coli O157:H7 by 64.29, 65.98, 70.41, and 85.59%, respectively. Furthermore, these concentrations inhibited 6.13, 45.50, 49.45, and 100%, respectively, of the Salmonella Typhimurium JSG 1748 biofilm. A mutagenicity assay confirmed that green cardamom essential oil has no demonstrable mutagenic activity against the tested strains. The study’s findings suggest that green cardamom derived bioactive compounds are safe organic antimicrobials, effective in controlling biofilm formation by Gram-negative pathogens. Moreover, such compounds could possibly be used in the food industry (e.g., bakery, dairy, meat, and other food products) as a safe alternative to chemical preservatives (antimicrobials) to enhance shelf life by improving the antimicrobial status while at the same time imparting a pleasant and appealing aroma for consumers.
The Heat Shock Transcription Factor HsfA Is Essential for Thermotolerance and Regulates Cell Wall Integrity in Aspergillus fumigatus
Authors: Fabri João Henrique Tadini Marilhano , Rocha Marina Campos , Fernandes Caroline Mota , Persinoti Gabriela Felix , Ries Laure Nicolas Annick , Cunha Anderson Ferreira da , Goldman Gustavo Henrique , Del Poeta Maurizio , Malavazi Iran
Published: April 2021

The deleterious effects of human-induced climate change have long been predicted. However, the imminent emergence and spread of new diseases, including fungal infections through the rise of thermotolerant strains, is still neglected, despite being a potential consequence of global warming. Thermotolerance is a remarkable virulence attribute of the mold Aspergillus fumigatus. Under high-temperature stress, opportunistic fungal pathogens deploy an adaptive mechanism known as heat shock (HS) response controlled by heat shock transcription factors (HSFs). In eukaryotes, HSFs regulate the expression of several heat shock proteins (HSPs), such as the chaperone Hsp90, which is part of the cellular program for heat adaptation and a direct target of HSFs. We recently observed that the perturbation in cell wall integrity (CWI) causes concomitant susceptibility to elevated temperatures in A. fumigatus, although the mechanisms underpinning the HS response and CWI cross talking are not elucidated. Here, we aim at further deciphering the interplay between HS and CWI. Our results show that cell wall ultrastructure is severely modified when A. fumigatus is exposed to HS. We identify the transcription factor HsfA as essential for A. fumigatus viability, thermotolerance, and CWI. Indeed, HS and cell wall stress trigger the coordinated expression of both hsfA and hsp90. Furthermore, the CWI signaling pathway components PkcA and MpkA were shown to be important for HsfA and Hsp90 expression in the A. fumigatus biofilms. Lastly, RNA-sequencing confirmed that hsfA regulates the expression of genes related to the HS response, cell wall biosynthesis and remodeling, and lipid homeostasis. Our studies collectively demonstrate the connection between the HS and the CWI pathway, with HsfA playing a crucial role in this cross-pathway regulation, reinforcing the importance of the cell wall in A. fumigatus thermophily.
Rock-Hosted Subsurface Biofilms: Mineral Selectivity Drives Hotspots for Intraterrestrial Life
Authors: Casar Caitlin P. , Kruger Brittany R. , Osburn Magdalena R.
Published: April 2021

The continental deep subsurface is likely the largest reservoir of biofilm-based microbial biomass on Earth, but the role of mineral selectivity in regulating its distribution and diversity is unclear. Minerals can produce hotspots for intraterrestrial life by locally enhancing biofilm biomass. Metabolic transformations of minerals by subsurface biofilms may occur widely with the potential to significantly impact subsurface biogeochemical cycles. However, the degree of impact depends upon the amount of biofilm biomass and its relationship to host rock mineralogy, estimates that are currently loosely constrained to non-existent. Here, we use in situ cultivation of biofilms on native rocks and coupled microscopy/spectroscopy to constrain mineral selectivity by biofilms in a deep continental subsurface setting: the Deep Mine Microbial Observatory (DeMMO). Through hotspot analysis and spatial modeling approaches we find that mineral distributions, particularly those putatively metabolized by microbes, indeed drive biofilm distribution at DeMMO, and that bioleaching of pyrite may be a volumetrically important process influencing fluid geochemistry at this site when considered at the kilometer scale. Given the ubiquity of iron-bearing minerals at this site and globally, and the amount of biomass they can support, we posit that rock-hosted biofilms likely contribute significantly to subsurface biogeochemical cycles. As more data becomes available, future efforts to estimate biomass in the continental subsurface should incorporate host rock mineralogy.
Age-Related Memory Impairment and Sex-Specific Alterations in Phosphorylation of the Rpt6 Proteasome Subunit and Polyubiquitination in the Basolateral Amygdala and Medial Prefrontal Cortex
Authors: Dulka Brooke N. , Trask Sydney , Helmstetter Fred J.
Published: April 2021

Aging is marked by an accumulation of damaged and modified brain proteins, and the ubiquitin-proteasome system (UPS) is important for cellular protein degradation. Recent work has established a critical role for the UPS in memory and synaptic plasticity, but the role of the UPS in age-related cognitive decline remains poorly understood. We trained young, middle-aged, and aged male and female rats using trace fear conditioning (TFC) to investigate the effects of age and sex on memory. We then measured markers of UPS-related protein degradation (phosphorylation of the Rpt6 proteasome regulatory subunit and K48-linked polyubiquitination) using western blots. We found that aged males, but not aged females, showed behavioral deficits at memory retrieval. Aged males also displayed reduced phosphorylation of the Rpt6 proteasome subunit and accumulation of K48 in the basolateral amygdala, while aged females displayed a similar pattern in the medial prefrontal cortex. These findings suggest that markers of UPS function are differentially affected by age and sex in a brain region-dependent manner. Together these results provide an important step toward understanding the UPS and circuit-level differences in aging males and females.
Convergent and Divergent Mechanisms of Epileptogenesis in mTORopathies
Authors: Nguyen Lena H. , Bordey Angélique
Published: April 2021

Hyperactivation of the mechanistic target of rapamycin complex 1 (mTORC1) due to mutations in genes along the PI3K-mTOR pathway and the GATOR1 complex causes a spectrum of neurodevelopmental disorders (termed mTORopathies) associated with malformation of cortical development and intractable epilepsy. Despite these gene variants’ converging impact on mTORC1 activity, emerging findings suggest that these variants contribute to epilepsy through both mTORC1-dependent and -independent mechanisms. Here, we review the literature on in utero electroporation-based animal models of mTORopathies, which recapitulate the brain mosaic pattern of mTORC1 hyperactivity, and compare the effects of distinct PI3K-mTOR pathway and GATOR1 complex gene variants on cortical development and epilepsy. We report the outcomes on cortical pyramidal neuronal placement, morphology, and electrophysiological phenotypes, and discuss some of the converging and diverging mechanisms responsible for these alterations and their contribution to epileptogenesis. We also discuss potential therapeutic strategies for epilepsy, beyond mTORC1 inhibition with rapamycin or everolimus, that could offer personalized medicine based on the gene variant.
Acetylcholine Receptor Stimulation Activates Protein Kinase C Mediated Internalization of the Dopamine Transporter
Authors: Underhill Suzanne M. , Amara Susan G.
Published: April 2021

The dopamine transporter (DAT) clears neurotransmitters from the extracellular space and serves as an important regulator of signal amplitude and duration at sites of dopamine release. Several different intracellular signaling pathways have been observed to modulate DAT activity through the regulation of the trafficking of the carriers to and from the cell surface. Acute activation of protein kinase C (PKC) by phorbol esters facilitates clathrin-dependent internalization of the DAT in a variety of model systems; however, the physiological stimuli and cell-surface receptor systems that activate PKC and regulate the DAT in dopamine neurons remain elusive. We report here that stimulation of M1/M5 muscarinic receptors in midbrain cultures decreases the ability of dopamine neurons to transport dopamine through DAT. Application of the cholinomimetic drug carbachol leads to a decrease in DAT activity in primary cultures while the M1/M5-specific antagonist, pirenzepine, blocks these effects. The M3 antagonist, DAU 5884, does not affect, but a positive modulator of M5, VU 0238429, enhances the loss of DAT function in response to carbachol and acetylcholine. These data implicate M1/M5 receptors on dopamine neurons in the modulation of DAT function. Bisindolylmaleimide, a PKC inhibitor, blocks the effects of carbachol stimulation on dopamine uptake, supporting a role for PKC in muscarinic receptor-mediated DAT internalization. Furthermore, as shown previously for PKC-induced internalization, downregulation of the DAT is dependent on both clathrin and dynamin. A Gq-specific inhibitor peptide also blocks the effects of carbachol on DAT in primary cultures, confirming Gq as the G-protein that couples M1/M5 receptors to PKC activation in these cells. In acute midbrain slices, biotinylation of cell-surface proteins revealed the loss of dopamine transport mediated by muscarinic receptor stimulation was, indeed, due to loss of membrane expression of the DAT in endogenous tissue. These data indicate that stimulation of cholinergic pathways can lead to modulation of dopamine through internalization of the DAT.
Prolonged Intubation in Patients With Prior Cerebrovascular Disease and COVID-19
Authors: Mukerji Shibani S. , Das Sudeshna , Alabsi Haitham , Brenner Laura N. , Jain Aayushee , Magdamo Colin , Collens Sarah I. , Ye Elissa , Keller Kiana , Boutros Christine L. , Leone Michael J. , Newhouse Amy , Foy Brody , Li Matthew D. , Lang Min , Anahtar Melis N. , Shao Yu-Ping , Ge Wendong , Sun Haoqi , Triant Virginia A. , Kalpathy-Cramer Jayashree , Higgins John , Rosand Jonathan , Robbins Gregory K. , Westover M. Brandon
Published: April 2021

Objectives: Patients with comorbidities are at increased risk for poor outcomes in COVID-19, yet data on patients with prior neurological disease remains limited. Our objective was to determine the odds of critical illness and duration of mechanical ventilation in patients with prior cerebrovascular disease and COVID-19.

Methods: A observational study of 1,128 consecutive adult patients admitted to an academic center in Boston, Massachusetts, and diagnosed with laboratory-confirmed COVID-19. We tested the association between prior cerebrovascular disease and critical illness, defined as mechanical ventilation (MV) or death by day 28, using logistic regression with inverse probability weighting of the propensity score. Among intubated patients, we estimated the cumulative incidence of successful extubation without death over 45 days using competing risk analysis.

Results: Of the 1,128 adults with COVID-19, 350 (36%) were critically ill by day 28. The median age of patients was 59 years (SD: 18 years) and 640 (57%) were men. As of June 2nd, 2020, 127 (11%) patients had died. A total of 177 patients (16%) had a prior cerebrovascular disease. Prior cerebrovascular disease was significantly associated with critical illness (OR = 1.54, 95% CI = 1.14–2.07), lower rate of successful extubation (cause-specific HR = 0.57, 95% CI = 0.33–0.98), and increased duration of intubation (restricted mean time difference = 4.02 days, 95% CI = 0.34–10.92) compared to patients without cerebrovascular disease.

Interpretation: Prior cerebrovascular disease adversely affects COVID-19 outcomes in hospitalized patients. Further study is required to determine if this subpopulation requires closer monitoring for disease progression during COVID-19.
Gene-Environment Interactions in Progressive Supranuclear Palsy
Authors: Litvan Irene , Proudfoot James A. , Martin Eden R. , Standaert David , Riley David , Hall Deborah , Marras Connie , Bayram Ece , Dubinsky Richard M. , Bordelon Yvette , Reich Stephen , Shprecher David , Kluger Benzi , Cunningham Christopher , Schellenberg Gerard D. , Jankovic Joseph
Published: April 2021

Several genetic and environmental factors have been reported in progressive supranuclear palsy (PSP), although none were identified as a definitive cause. We aimed to explore potential gene-environment interactions in PSP. Two hundred and ninety two PSP cases and 292 controls matched for age, sex, and race from the ENGENE-PSP were analyzed to determine the association between PSP and minor alleles of 5 single nucleotide polymorphisms (SNPs) in 4 genes (MAPT, MOBP, EIF2AK3, and STX6), which were previously associated with PSP risk. Interactions between these SNPs and environmental factors, including previously reported occupational and agricultural risk factors for PSP, were assessed for PSP odds and age of symptom onset. Minor alleles of MAPTrs242557 and EIF2AK3rs7571971 were individually associated with increased odds; MAPTrs8070723 minor alleles were associated with lower PSP odds. There were several gene-environment interactions for PSP odds and age of symptom onset, however, they did not remain significant after FDR-correction. Larger scale studies are required to determine potential interactions.
Dietary Quality and Sociodemographic and Health Behavior Characteristics Among Pregnant Women Participating in the New York University Children's Health and Environment Study
Authors: Deierlein Andrea L. , Ghassabian Akhgar , Kahn Linda G. , Afanasyeva Yelena , Mehta-Lee Shilpi S. , Brubaker Sara G. , Trasande Leonardo
Published: April 2021

Maternal diet, prior to and during pregnancy, plays an important role in the immediate and long-term health of the mother and her offspring. Our objectives were to assess diet quality among a large, diverse, urban cohort of pregnant women, and examine associations with sociodemographic and health behavior characteristics. Data were from 1,325 pregnant women enrolled in New York University Children's Health and Environment Study (NYU CHES). Diet quality was assessed using the Healthy Eating Index (HEI)-2015. Mean total HEI-2015 score was 74.9 (SD = 8.5); 376 (28%), 612 (46%), 263 (20%), and 74 (6%) of women had scores that fell into the grade range of A/B, C, D, and F, respectively. Mean HEI-2015 component scores were high for fruit and whole grains and low for protein-related, sodium, and fat-related components. In multivariable linear regression models, Hispanic women scored 1.65 points higher on the total HEI-2015 (95% CI: 0.21, 3.10) compared to non-Hispanic White women, while younger age (<30 years), parity, single status, pre-pregnancy obesity, smoking, pre-existing hypertension, moderate/severe depressive symptoms, not meeting physical activity recommendations, and not taking a vitamin before pregnancy were associated with ~1.5–5-point lower mean total HEI-2015 scores. Diet is a modifiable behavior; our results suggest a continued need for pre-conceptional and prenatal nutritional counseling.
Molecular Chaperone GRP94/GP96 in Cancers: Oncogenesis and Therapeutic Target
Authors: Duan Xiaofeng , Iwanowycz Stephen , Ngoi Soo , Hill Megan , Zhao Qiang , Liu Bei
Published: April 2021

During tumor development and progression, intrinsic and extrinsic factors trigger endoplasmic reticulum (ER) stress and the unfolded protein response, resulting in the increased expression of molecular chaperones to cope with the stress and maintain tumor cell survival. Heat shock protein (HSP) GRP94, also known as GP96, is an ER paralog of HSP90 and has been shown to promote survival signaling during tumor-induced stress and modulate the immune response through its multiple clients, including TLRs, integrins, LRP6, GARP, IGF, and HER2. Clinically, elevated expression of GRP94 correlates with an aggressive phenotype and poor clinical outcome in a variety of cancers. Thus, GRP94 is a potential molecular marker and therapeutic target in malignancies. In this review, we will undergo deep molecular profiling of GRP94 in tumor development and summarize the individual roles of GRP94 in common cancers, including breast cancer, colon cancer, lung cancer, liver cancer, multiple myeloma, and others. Finally, we will briefly review the therapeutic potential of selectively targeting GRP94 for the treatment of cancers.
Challenges and Initiatives in Diversity, Equity and Inclusion in Cancer Molecular Imaging
Authors: Daldrup-Link Heike E. , Esposito Giuseppe , Bhujwalla Zaver M.
Published: April 2021

A diverse biomedical workforce is essential to achieve excellence in patient care, clinical translational, and basic research. Diversity, equity, and inclusion challenges in cancer molecular represent a combination of the challenges facing the science, technology, engineering, and mathematics (STEM) field, and challenges in Radiology and Nuclear Medicine. Although there is a growing awareness of conscious and unconscious bias that negatively affect the cancer imaging world, many challenges remain such as overcoming barriers to entry into the pipeline, avoiding program dropout, and providing long-term career prospect. The COVID-19 pandemic has resulted in a significant setback and further highlighted problems faced by women and underrepresented minorities. In this perspective, we have identified some of the challenges faced and highlighted ongoing and future initiatives to address these challenges.
Effects of Hydroxylated Mephedrone Metabolites on Monoamine Transporter Activity in vitro
Authors: Niello Marco , Cintulová Daniela , Raithmayr Philip , Holy Marion , Jäntsch Kathrin , Colas Claire , Ecker Gerhard F. , Sitte Harald H. , Mihovilovic Marko D.
Published: April 2021

Mephedrone is a largely abused psychostimulant. It elicits the release of monoamines via the high affinity transporters for dopamine (DAT), norepinephrine (NET) and serotonin (SERT). Stereoselective metabolic reactions are involved in the inactivation and the elimination of its chemical structure. However, during these processes, several structures are generated and some of them have been reported to be still pharmacologically active. In this study 1) we have newly synthetized several putative mephedrone metabolites, 2) compared their activity at monoamine transporters, 3) generated quantitative structure activity relationships, and 4) exploited the chemical structure of the putative metabolites to screen a urine sample from a drug user and dissect mephedrone metabolism. We have found that most of the tested metabolites are weak inhibitors of monoamine transporters and that all of them are more potent at DAT and NET in comparison to SERT. The only exception is represented by the COOH-metabolite which shows no pharmacological activity at all three monoamine transporters. The enantioselectivity of mephedrone and its metabolites is present mainly at SERT, with only minor effects at DAT and NET being introduced when the β-keto group is reduced to an OH-group. Importantly, while at DAT the putative metabolites did not show changes in inhibitory potencies, but rather changes in their substrate/blocker profile, at SERT they showed mainly changes in inhibitory potencies. Molecular modeling suggests that the hydrophobic nature of a specific SERT subpocket may be involved in such loss of affinity. Finally, the assessment of the putative metabolites in one urine sample of mephedrone user displayed two previously uncharacterized metabolites, 4-COOH-nor-mephedrone (4-COOH-MC) and dihydro-4- nor-mephedrone (dihydro-4-MC). These results confirm and expand previous studies highlighting the importance of the stereochemistry in the pharmacodynamics of phase-1 metabolites of mephedrone, established their structure-activity relationships at DAT, NET and SERT and pave the way for a systematic dissection of mephedrone metabolic routes. Given the number of structures found having residual and modified pharmacological profiles, these findings may help in understanding the complex subjective effects of administered mephedrone. Moreover, the dissection of mephedrone metabolic routes may help in developing new therapies for treating psychostimulants acute intoxications.
Microbial Metabolite Sodium Butyrate Attenuates Cartilage Degradation by Restoring Impaired Autophagy and Autophagic Flux in Osteoarthritis Development
Authors: Zhou Haikang , Li Guoqing , Wang Yang , Jiang Rendong , Li Yicheng , Wang Huhu , Wang Fei , Ma Hairong , Cao Li
Published: April 2021

Osteoarthritis (OA) is a degenerative joint disease with multiple etiologies that affects individuals worldwide. No effective interventions are currently available to reverse the pathological process of OA. Sodium butyrate (NaB), a component of short-chain fatty acids (SCFAs), has multiple biological activities, including the attenuation of inflammation and anti-tumor activities in various diseases. However, whether the protective effects of NaB in OA are associated with the promotion of autophagy had not been investigated. Here, we explored the chondroprotective properties of NaB in an interleukin (IL)-1β-induced inflammatory chondrocyte model and an anterior cruciate ligament transection (ACLT) mouse model. Hematoxylin and eosin (HE), Safranin O, and immunohistochemical staining were performed to evaluate the effects of NaB treatment on articular cartilage. An optimal NaB dose for chondrocyte treatment was determined via cell counting kit-8 assays. Immunofluorescence and transmission electron microscopy were used to detect autophagy in chondrocytes. Flow cytometry was utilized to detect reactive oxygen species (ROS), cell cycle activity, and apoptosis in chondrocytes. Western blot and immunostaining were performed to evaluate the protein expression levels of relevant indicators. We found that the administration of NaB by oral gavage could attenuate cartilage degradation. In parallel, NaB treatment could enhance the activation of autophagy, increase autophagic flux, decrease extracellular matrix degradation, and reduce apoptosis by restraining inflammation, ROS production, and cell cycle arrest in IL-1β-treated chondrocytes. The protective effects of NaB could be partially abolished by the autophagy inhibitor 3-methyladenine (3-MA), which indicated that the protective effects of NaB against OA were partially governed by the enhancement of autophagy to restrain the formation of inflammatory mediators and ROS and regulate cell cycle progression and apoptosis in chondrocytes. In conclusion, NaB could attenuate OA progression by restoring impaired autophagy and autophagic flux via the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) pathway, both in vitro and in vivo, implying that NaB could represent a novel therapeutic approach for OA.
Characterization of Glycoside Hydrolase Families 13 and 31 Reveals Expansion and Diversification of α-Amylase Genes in the Phlebotomine Lutzomyia longipalpis and Modulation of Sandfly Glycosidase Activities by Leishmania Infection
Authors: da Costa-Latgé Samara Graciane , Bates Paul , Dillon Rod , Genta Fernando Ariel
Published: April 2021

Sugar-rich food sources are essential for sandflies to meet their energy demands, achieving more prolonged survival. The digestion of carbohydrates from food is mainly realized by glycoside hydrolases (GH). To identify genes coding for α-glycosidases and α-amylases belonging to Glycoside Hydrolase Family 13 (GH13) and Glycoside Hydrolase Family 31 (GH31) in Lutzomyia longipalpis, we performed an HMMER search against its genome using known sequences from other dipteran species. The sequences retrieved were classified based on BLASTP best hit, analysis of conserved regions by alignment with sequences of proteins with known structure, and phylogenetic analysis comparing with orthologous proteins from other dipteran species. Using RT-PCR analysis, we evaluated the expression of GH13 and GH31 genes, in the gut and rest of the body of females, in four different conditions: non-fed, sugar-fed, blood-fed, and Leishmania mexicana infected females. L. longipalpis has GH13/31 genes that code for enzymes involved in various aspects of sugar metabolism, as carbohydrate digestion, storage, and mobilization of glycogen reserves, proteins involved in transport, control of N-glycosylation quality, as well as others with a putative function in the regulation of myogenesis. These proteins are representatives of GH13 and GH31 families, and their roles seem to be conserved. Most of the enzymes seem to be active with conserved consense sequences, including the expected catalytic residues. α-amylases also demonstrated the presence of calcium and chloride binding sites. L. longipalpis genome shows an expansion in the α-amylase gene family, with two clusters. In contrast, a retraction in the number of α-glucosidases occurred. The expansion of α-amylases is probably related to the specialization of these proteins for different substrates or inhibitors, which might correlate with the higher diversity of plant foods available in the natural habitat of L. longipalpis. The expression of α-glucosidase genes is higher in blood-fed females, suggesting their role in blood digestion. Besides that, in blood-fed females infected with the parasite Leishmania mexicana, these genes were also modulated. Glycoside Hydrolases from families 13 and 31 are essential for the metabolism of L. longipalpis, and GH13 enzymes seem to be involved in the interaction between sandflies and Leishmania.
Vulnerable Plaque Is More Prevalent in Male Individuals at High Risk of Stroke: A Propensity Score-Matched Study
Authors: Li Jie , Gao Lijie , Zhang Ping , Liu Yingying , Zhou Ju , Yi Xingyang , Wang Chun
Published: April 2021

Objectives

To assess the gender differences in the prevalence of carotid vulnerable plaques in high-risk individuals for stroke in a multicenter, cross-sectional study.

Methods

In the year 2015, 18595 residents who were at the age of 40 or older participated in a face-to-face study in eight communities in southwestern China. Totally 2,644 participants at high risk of stroke were enrolled. Before and after propensity score matching (PSM), the prevalence of carotid plaques and vulnerable plaques were compared between men and women. Multivariate analyses were applied to explore the association between the gender and carotid plaques. Stratified analyses and interaction tests were performed to identify factors that might modify the association between the gender and carotid plaques.

Results

Among 2644 high-risk individuals enrolled, there were 1,202 (45.5%) men and 1442 (54.5%) women. Carotid plaques were detected in 904 (34.2%) participants, while vulnerable plaques were found in 425 (16.1%) participants. Before PSM, carotid plaques were more prevalent in male individuals than the female (36.7% vs. 32.1%, p = 0.01), as well as vulnerable plaque (20.0% vs. 12.8%, p < 0.01). Men tend to have a higher prevalence of vulnerable plaques in multivariate analyses (adjusted OR 1.70, 95% CI 1.10–2.62, p = 0.02). Stratified analyses and interaction tests demonstrated that the association between male sex and vulnerable carotid plaque did not change by age, family history of stroke, histories of chronic disease, smoking status, drinking status, physical activity, and BMI (all p for interaction > 0.05). After PSM, vulnerable plaques were still more prevalent in male individuals than the female (17.03% vs. 12.07%, p = 0.032).

Conclusion

Male individuals had a higher risk of vulnerable carotid plaque independent of classical vascular risk factors. Whether there is a gender-specific association between variations in genes related to inflammation, lipid metabolis, and endothelial function and plaque vulnerability needs to be further studied.