bluuebunny/crossref_metadata_2025 huggingface dataset:
Optic Nerve Angle in Idiopathic Intracranial Hypertension
Authors: Chen Benson S. , Asnafi Solmaz , Lin Mung Y. , Bruce Beau B. , Lock Jane H. , Sharma Rahul A. , Newman Nancy J. , Biousse Valérie , Saindane Amit M.
Published: June 2020

Background:

The tortuosity of the optic nerve can be quantified radiologically by measuring the angle of optic nerve deformation (the “optic nerve angle” [ONA]). In patients with idiopathic intracranial hypertension (IIH), lowering the intracranial pressure (ICP) to a normal range by lumbar puncture leads to straightening of the optic nerve and an increase in the measured sagittal ONA on MRI. It is uncertain whether there is any correlation between ONA and cerebrospinal fluid (CSF) opening pressure or visual function.

Methods:

Retrospective study of patients with and without IIH who had MRI of the brain followed by lumbar puncture with CSF opening pressure within 24 hours of MRI. Before LP and within 24 hours of MRI of the brain, all patients with IIH had neuro-ophthalmologic assessment including visual acuity, Humphrey Visual Field (HVF), and fundus photography. Sagittal ONA was measured on multiplanar T2-SPACE images on a DICOM viewer. Papilledema on the fundus photographs was graded using the Frisén scale.

Results:

Fifty-four patients with IIH and 30 unmatched controls were included. The IIH group was 6.3 years younger (95% CI 2.4–10.3, P = 0.002), had 8.7 kg/m2 higher body mass index (4.9–12.5, P < 0.001), and 26.3% more women (P = 0.011) compared with controls. In both eyes, the ONA was significantly smaller in patients with IIH by 12° compared with controls (7°–17°, P < 0.00001). In the IIH group, no correlation between ONA and the CSF opening pressure was present in either eye (right eye r = 0.19, P = 0.15; left eye r = 0.18, P = 0.19) The ONA did not correlate with logarithm of the minimum angle of resolution visual acuity (right eye r = 0.26, P = 0.063; left eye r = 0.15, P = 0.27), HVF mean deviation (right eye r = 0.0059, P = 0.97; left eye r = −0.069, P = 0.63), or Frisén grade (Spearman's rho right eye 0.058, P = 0.67; left eye 0.14, P = 0.30).

Conclusions:

The ONA is significantly smaller in patients with IIH compared to controls, but does not correlate with CSF opening pressure, severity of papilledema, or visual function. The ONA may be useful in identifying patients with raised ICP, but not necessarily those with a poor visual prognosis.
Progressive Functional and Neuroretinal Affectation in Patients With Multiple Sclerosis Treated With Fingolimod
Authors: Garcia-Martin Elena , Ruiz de Gopegui Erika , Satue Maria , Gil-Arribas Laura , Jarauta Laura , Ara Jose R. , Martin Jesus , Fernandez Francisco J. , Vilades Elisa , Rodrigo Maria J.
Published: June 2020

Background:

To evaluate the effect of fingolimod in visual function and neuroretinal structures in patients with multiple sclerosis (MS) for a period of 1 year.

Methods:

This longitudinal and observational cohort study included 78 eyes of 78 patients with MS treated with fingolimod. All subjects were evaluated every 3 months during 12 months and compared with 32 patients treated with interferon beta. All patients were examined for high-contrast and low-contrast (2.5% and 1.25%) visual acuity (VA), contrast sensitivity vision (CSV) (using Pelli–Robson and CSV-1000E tests), color vision (Farnsworth D-15 and L'Anthony D-15 desaturated tests), and retinal structural measurements (retinal nerve fiber layer [RNFL] and ganglion cell layer [GCL] thickness) using optical coherence tomography (OCT) technology.

Results:

Patients with MS treated with fingolimod for a period of 1 year showed significant reduction in 100% and 1.25% contrast VA (P = 0.009 and 0.008, respectively), an alteration of contrast sensitivity and color perception (Pelli–Robson test, CSV-1000E test, Farnsworth D-15 desaturated test, and L'Anthony D-15 desaturated test; P < 0.001), GCL thickness reduction (P = 0.007), and an average macular central thickness increase of 2.6 μm (P = 0.006). Patients with MS treated with interferon beta did not show significant changes in visual function tests neither in macular thickness measurements, but they showed a significant reduction in GCL and RNFL thicknesses. The reduction in neuroretinal structures observed by OCT was significantly higher in the interferon-beta group, but patients treated with fingolimod showed a significant increase in macular central thickness and a reduction in low contrast vision (P < 0.001).

Conclusions:

Patients with MS treated with fingolimod and with no clinically observable macular edema show a significant change in visual function parameters and average macular central thickness increase compared with those treated with interferon beta. These findings are probably due to subclinical macular edema produced by fingolimod, which might be considered as an indicator for pharmacovigilance of sphingosine-1-phosphate inhibitors to be improved.
Cyclotorsion Measurement on Scanning Laser Ophthalmoscopy Imaging Compared With Fundus Photography in Patients With Fourth Nerve Palsy and Healthy Controls: CySLO-IV Study
Authors: Kanku Madeleine S. , Sele Silvano , Held Ulrike , Jaggi Gregor P. , Landau Klara , Traber Ghislaine L.
Published: June 2020

Background:

The purpose of the study is to evaluate the agreement of the foveopapillary angle (FPA) on conventional fundus photography (c-FPA) with the FPA on scanning laser ophthalmoscopy (SLO) imaging (SLO-FPA) in patients with fourth nerve palsy and healthy controls (HCs).

Methods:

The FPA was measured in both eyes of 25 patients and 25 HCs in synedra View (c-FPA) and with the integrated algorithm of the Heidelberg Spectralis OCT (SLO-FPA). The primary endpoint was the agreement of both measurements. Furthermore, we evaluated the influence of the eye tracker, the influence of fixation on objective torsion, and the FPA cutoff between patients and HCs.

Results:

The mean SLO-FPA in patients (6/25 acquired palsies) was 11.3 ± 3.6° and 6.4 ± 2.1° in HCs. The mean c-FPA was 11.4 ± 4.0° and 5.8 ± 2.2°, respectively. The Bland–Altman plot of c-FPA vs SLO-FPA in patients and HCs shows no systematic bias (mean of −0.28°). Limits of agreement were −6.58 and 6.02°. Using the eye tracker had no systematic effect. There was no evidence for an immediate shift of torsion with change of fixation (24/25 patients and 23/25 HCs). Discrimination between patients and HCs by the SLO-FPA is very good with an area under the curve = 0.92 (95% confidence interval: 0.84–0.99).

Conclusions:

SLO-FPA measurement allows convenient and consistent assessment of objective cyclotorsion. There was no systematic bias in the difference between SLO-FPA and traditional c-FPA; thus, SLO-FPA is a valuable alternative to the commonly used c-FPA. Using the eye tracker is recommended for proper centering of the ring scan.
Idiopathic Intracranial Hypertension: Glymphedema of the Brain
Authors: Nicholson Patrick , Kedra Alice , Shotar Eimad , Bonnin Sophie , Boch Anne-Laure , Shor Natalia , Clarençon Frédéric , Touitou Valérie , Lenck Stephanie
Published: June 2020

Background:

During the last decade, our understanding of cerebrospinal fluid (CSF) physiology has dramatically improved, thanks to the discoveries of both the glymphatic system and lymphatic vessels lining the dura mater in human brains.

Evidence Acquisition:

We detail the recent basic science findings in the field of CSF physiology and connect them with our current understanding of the pathophysiology of idiopathic intracranial hypertension (IIH).

Results:

Transverse sinus (TS) stenoses seem to play a major causative role in the symptoms of IIH, as a result of a decrease in the pressure gradient between the venous system and the subarachnoid space. However, the intracranial pressure can be highly variable among different patients, depending on the efficiency of the lymphatic system to resorb the CSF and on the severity of TS stenoses. It is likely that there is a subclinical form of IIH and that IIH without papilledema is probably under-diagnosed among patients with chronic migraines or isolated tinnitus.

Conclusions:

IIH can be summarized in the following pathological triad: restriction of the venous CSF outflow pathway—overflow of the lymphatic CSF outflow pathway—congestion of the glymphatic system. To better encompass all the stages of IIH, it is likely that the Dandy criteria need to be updated and that perhaps renaming IIH should be considered.
Saliva sampling is an excellent option to increase the number of SARS CoV2 diagnostic tests in settings with supply shortages
Authors: Moreno-Contreras Joaquín , Espinoza Marco A. , Sandoval-Jaime Carlos , Cantú-Cuevas Marco A. , Barón-Olivares Héctor , Ortiz-Orozco Oscar D. , Muñoz-Rangel Asunción V. , Hernández-de la Cruz Manuel , Eroza-Osorio César M. , Arias Carlos F. , López Susana
Published: June 2020

As part of any plan to lift or ease the confinement restrictions that are in place in many different countries, there is an urgent need to increase the capacity of laboratory testing for SARS CoV-2. Detection of the viral genome through RT-qPCR is the golden standard for this test, however, the high demand of the materials and reagents needed to sample individuals, purify the viral RNA, and perform the RT-qPCR test has resulted in a worldwide shortage of several of these supplies. Here, we show that directly lysed saliva samples can serve as a suitable source for viral RNA detection that is cheaper and can be as efficient as the classical protocol that involves column purification of the viral RNA. In addition, it surpasses the need for swab sampling, decreases the risk of the healthcare personnel involved in this process, and accelerates the diagnostic procedure.
Increased axon initial segment length results in increased Na+currents in spinal motoneurones at symptom onset in the G127X SOD1 mouse model of Amyotrophic Lateral Sclerosis
Authors: Jørgensen H. S. , Jensen D.B. , Dimintiyanova K.P. , Bonnevie V.S. , Hedegaard A. , Lehnhoff J. , Moldovan M. , Grondahl L. , Meehan C.F.
Published: June 2020

Amyotrophic lateral sclerosis is a neurodegenerative disease preferentially affecting motoneurones. Transgenic mouse models have been used to investigate the role of abnormal motoneurone excitability in this disease. Whilst an increased excitability has repeatedly been demonstratedin vitroin neonatal and embryonic preparations from SOD1 mouse models, the results from the only studies to recordin vivofrom spinal motoneurones in adult SOD1 models have produced conflicting findings. Deficits in repetitive firing have been reported in G93A SOD1 mice but not in presymptomatic G127X SOD1 mice despite shorter motoneurone axon initial segments (AISs) in these mice.

These discrepancies may be due to the earlier disease onset and prolonged disease progression in G93A SOD1 mice with recordings potentially performed at a later sub-clinical stage of the disease in this mouse. To test this, and to explore how the evolution of excitability changes with symptom onset we performedin vivointracellular recording and AIS labelling in G127X SOD1 mice immediately after symptom onset. No reductions in repetitive firing were observed showing that this is not a common feature across all ALS models. Immunohistochemistry for the Na+channel Nav1.6 showed that motoneurone AISs increase in length in G127X SOD1 mice at symptom onset. Consistent with this, the rate of rise of AIS components of antidromic action potentials were significantly faster confirming that this increase in length represents an increase in AIS Na+channels occurring at symptom onset in this model.

Highights

In vivoelectrophysiological recordings were made in symptomatic G127X SOD1 mice.

There were no deficits in repetitive firing in motoneurones in G127X mice.

Increased persistent inward currents were still present in the symptomatic mice.

Results suggest increases in Na+currents at axon initial segments (AISs).

Immunohistochemistry showed that motoneurone AISs were longer and thinner.
Checkpoint phosphorylation sites on budding yeast Rif1 protect nascent DNA from degradation by Sgs1-Dna2
Authors: Monerawela Chandre , Hiraga Shin-ichiro , Donaldson Anne D.
Published: June 2020

In budding yeast the Rif1 protein is important for protecting nascent DNA at blocked replication forks, but the mechanism has been unclear. Here we show that budding yeast Rif1 must interact with Protein Phosphatase 1 to protect nascent DNA. In the absence of Rif1, removal of either Dna2 or Sgs1 prevents nascent DNA degradation, implying that Rif1 protects nascent DNA by targeting Protein Phosphatase 1 to oppose degradation by the Sgs1-Dna2 nuclease-helicase complex. This functional role for Rif1 is conserved from yeast to human cells. Yeast Rif1 was previously identified as a target of phosphorylation by the Tel1/Mec1 checkpoint kinases, but the importance of this phosphorylation has been unclear. We find that nascent DNA protection depends on a cluster of Tel1/Mec1 consensus phosphorylation sites in the Rif1 protein sequence, indicating that the intra-S phase checkpoint acts to protect nascent DNA through Rif1 phosphorylation. Our observations uncover the pathway by which budding yeast Rif1 stabilises newly synthesised DNA, highlighting the crucial role Rif1 plays in maintaining genome stability from lower eukaryotes to humans.

Author summary

Genome instability is a leading factor contributing to cancer. Maintaining efficient error-free replication of the genome is key to preventing genome instability. During DNA replication, replication forks can be stalled by external and intrinsic obstacles, leading to processing of nascent DNA ends to enable replication restart. However, the nascent DNA must be protected from excessive processing to prevent terminal fork arrest, which could potentially lead to more serious consequences including failure to replicate some genome sequences. Using a nascent DNA protection assay we have investigated the role of the budding yeast Rif1 protein at blocked replication forks. We find that Rif1 protects nascent DNA through a mechanism that appears conserved from yeast to humans. We show that budding yeast Rif1 protects nascent DNA by targeting Protein Phosphatase 1 activity to prevent degradation of nascent DNA by the Sgs1-Dna2 helicase-nuclease complex. Furthermore, we find that Rif1 phosphorylation by the checkpoint pathway during replication stress is crucial for this function. Our results indicate that the S phase checkpoint machinery acts by phosphorylating Rif1 to protect nascent DNA, providing important clues concerning the conserved role of Rif1 in regulating events when replication is challenged.
Binding Ability Prediction between Spike Protein and Human ACE2 Reveals the Adaptive Strategy of SARS-CoV-2 in Humans
Authors: Xue Xia , Shi Jianxiang , Xu Hongen , Qin Yaping , Yang Zengguang , Feng Shuaisheng , Liu Danhua , Jian Liguo , Hua Linlin , Wang Yaohe , Zhang Qi , Huang Xueyong , Zhang Xiaoju , Li Xinxin , Chen Chunguang , Guo Jiancheng , Tang Wenxue , Liu Jianbo
Published: June 2020

SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) is a novel coronavirus causing an outbreak of COVID-19 globally in the past six months. A relatively higher divergence on the spike protein of SASR-CoV-2 enables it to transmit across species efficiently. We particularly believe that the adaptive mutations of the receptor-binding domain (RBD) of spike protein in SARS-CoV-2 might be essential to its high transmissibility among humans. Thus here we collected 2,142 high-quality genome sequences of SARS-CoV-2 from 160 regions in over 50 countries and reconstructed their phylogeny, and also analyzed the interaction between the polymorphisms of spike protein and human ACE2 (hACE2). Phylogenetic analysis of SARS-CoV-2 and coronavirus in other hosts show SARS-CoV-2 is highly possible originated from Bat-CoV (RaTG13) found in horseshoe bat and a recombination event may occur on the spike protein of Pangolin-CoV to imbue it the ability to infect humans. Moreover, compared to the S gene of SARS-CoV-2, it is more conserved in the direct-binding sites of RBD and we noticed that spike protein of SARS-CoV-2 may under a consensus evolution to adapt to human hosts better. 3,860 amino acid mutations in spike protein RBD (T333-C525) of SARS-CoV-2 were simulated and their stability and affinity binding to hACE2 (S19-D615) were calculated. Our analysis indicates SARS-CoV-2 could infect humans from different populations with no preference, and a higher divergence in the spike protein of SARS-CoV-2 at the early stage of this pandemic may be a good indicator that could show the pathway of SARS-CoV-2 transmitting from the natural reservoir to human beings.
Cis-regulatory mutations associate with transcriptional and post-transcriptional deregulation of the gene regulatory program in cancers
Authors: Castro-Mondragon Jaime A. , Ragle Aure Miriam , Lingjærde Ole Christian , Langerød Anita , Martens John W. M. , Børresen-Dale Anne-Lise , Kristensen Vessela , Mathelier Anthony
Published: June 2020

Background

Most cancer alterations occur in the noncoding portion of the human genome, which contains important regulatory regions acting as genetic switches to ensure gene expression occurs at correct times and intensities in correct tissues. However, large scale discovery of noncoding events altering the gene expression regulatory program has been limited to a few examples with high recurrence or high functional impact.

Results

We focused on transcription factor binding sites (TFBSs) that show similar mutation loads than what is observed in protein-coding exons. By combining cancer somatic mutations in TFBSs and expression data for protein-coding and miRNA genes, we evaluated the combined effects of transcriptional and post-transcriptional alteration on the dysregulation of the regulatory programs in cancer. The analysis of seven cancer cohorts culminated with the identification of protein-coding and miRNA genes linked to mutations at TFBSs that were associated with a cascading trans-effect deregulation on the cells’ regulatory program. Our analyses of cis-regulatory mutations associated with miRNAs recurrently predicted 17 miRNAs as pan-cancer-associated through deregulation of their target gene networks. Overall, our predictions were enriched for protein-coding and miRNA genes previously annotated as cancer drivers. Functional enrichment analyses highlighted that cis-regulatory mutations are associated with the dysregulation of key pathways associated with carcinogenesis

Conclusions

These pan-cancer results suggest that our method predicts cis-regulatory mutations related to the dysregulation of key gene regulatory networks in cancer patients. It highlights how the gene regulatory program is disrupted in cancer cells by combining transcriptional and post-transcriptional regulation of gene expression.
When do apples stop growing, and why does it matter?
Authors: Christodoulou Maria D , Culham Alastair
Published: June 2020

Background

Apples in the commercial food chain are harvested up to two weeks before maturity. We explore apple fruit development through the growing season to establish the point at which the features differentiating those cultivars become evident. This is relevant both for the understanding of the growing process and to ensure that any identification and classification tools can be used both on ripened-on-tree and stored fruit. Current literature presents some contradictory findings on apple development, we explored the size development of 12 apple cultivars in the Brogdale National Fruit Collection, UK over two growing seasons.

Methods

Fruit were sampled at regular time points throughout the growing season and four morphometrics (maximum length, maximum diameter, weight, and centroid size) were collected. These were regressed against growing degree days in order to appropriately describe the growth pattern observed.

Results

All four morphometrics were adequately described using log-log linear regressions, with adjusted R2estimates ranging from 78.3% (maximum length) to 86.7% (weight). For all four morphometrics, a 10% increase in growing degree days was associated with a 1% increase in the morphometric measurement.

Discussion

Our findings refine previous work presenting rapid early growth followed by a plateau in later stages of development and are in disagreement with published expo-linear models. We established that apples harvested for commercial storage purposes, two weeks prior to maturity, showed only a modest decrease in size, demonstrating that size morphometric approaches are appropriate for classification of apples, both ripened-on-tree and stored.
Expansin-controlled cell wall stiffness regulates root growth inArabidopsis
Authors: Samalova Marketa , Elsayad Kareem , Melnikava Alesia , Peaucelle Alexis , Gahurova Evelina , Gumulec Jaromir , Spyroglou Ioannis , Zemlyanskaya Elena V. , Ubogoeva Elena V. , Hejatko Jan
Published: June 2020

Expansins facilitate cell expansion via mediating pH-dependent cell wall (CW) loosening. However, the role of expansins in the control of biomechanical CW properties in the tissue and organ context remains elusive. We determined hormonal responsiveness and specificity of expression and localization of expansins predicted to be direct targets of cytokinin signalling. We found EXPA1 homogenously distributed throughout the CW of columella/ lateral root cap, while EXPA10 and EXPA14 localized predominantly at the three-cell boundaries of epidermis/cortex in various root zones. Cell type-specific localization of EXPA15 overlaps with higher CW stiffness measured via Brillouin light scattering microscopy. As indicated by both Brillouin frequency shift and AFM-measured Young’s modulus,EXPA1overexpression upregulated CW stiffness, associated with shortening of the root apical meristem and root growth arrest. We propose that root growth inArabidopsisrequires delicate orchestration of biomechanical CW properties via tight regulation of various expansins’ localization to specific cell types and extracellular domains.
Fi-score: a novel approach to characterise protein topology and aid in drug discovery studies
Authors: Kanapeckaitė Austė , Beaurivage Claudia , Hancock Matthew , Verschueren Erik
Published: June 2020

Target evaluation is at the centre of rational drug design and biologics development. In order to successfully engineer antibodies, T-cell receptors or small molecules it is necessary to identify and characterise potential binding or contact sites on therapeutically relevant target proteins. Currently, there are numerous challenges in achieving a better docking precision as well as characterising relevant sites. We devised a first-of-its-kindin silicoprotein fingerprinting approach based on dihedral angle and B-factor distribution to probe binding sites and sites of structural importance. In addition, we showed that the entire protein regions or individual structural subsets can be profiled using our derived fi-score based on amino acid dihedral angle and B-factor distribution. We further described a method to assess the structural profile and extract information on sites of importance using machine learning Gaussian mixture models. In combination, these biophysical analytical methods could potentially help to classify and systematically analyse not only targets but also drug candidates that bind to specific sites which would greatly improve pre-screening stage, target selection and drug repurposing efforts in finding other matching targets.
Dissection ofPlasmodium falciparumdevelopmental stages with multiple imaging methods
Authors: Preißinger Katharina , Vértessy Beáta , Kézsmárki István , Kellermayer Miklós
Published: June 2020

Efficient malaria treatment is a global challenge, requiring in-depth view into the maturation of malaria parasites during the intraerythrocytic cycle. Exploring structural and functional variations of the parasites through the intraerythrocytic stages and their impact on red blood cell (RBCs) is a cornerstone of antimalarial drug development. In order to trace such changes in fine steps of parasite development, we performed an imaging study of RBCs infected byPlasmodium falciparum, using atomic force microscopy (AFM) and total internal reflection fluorescence microscopy (TIRF), further supplemented with bright field microscopy for the direct assignment of the stages. This multifaceted imaging approach allows to reveal structure–functionality relations via correlations of the parasite maturation with morphological and fluorescence properties of the stages. We established identification patterns characteristic to the different parasite stages based on the height profile of infected RBCs, as obtained by AFM, which show close correlation with typical fluorescence (TIRF) maps of RBCs. Furthermore, we found that hemozoin crystals exhibit a strong optical contrast by quenching fluorescence. We demonstrate that these topographic and optical features also provide a tool to locate the hemozoin crystals within the RBCs and, in turn, to follow their growth.
The genome of North American beaver provides insights into the mechanisms of its longevity and cancer resistance
Authors: Zhang Quanwei , Tombline Gregory , Ablaeva Julia , Zhang Lei , Zhou Xuming , Smith Zachary , Xiaoli Alus M. , Wang Zhen , Lin Jhih-Rong , Jabalameli M. Reza , Mitra Joydeep , Nguyen Nha , Vijg Jan , Seluanov Andrei , Gladyshev Vadim N. , Gorbunova Vera , Zhang Zhengdong D.
Published: June 2020

The North American beaver (Castor canadensis) is an exceptionally long-lived and cancer-resistant rodent species, and thus an excellent model organism for comparative genomic studies of longevity. Here, we utilize a significantly improved beaver genome assembly to assess evolutionary changes in gene coding sequences, copy number, and expression. We found that the beaverAldh1a1, a stem cell marker gene encoding an enzyme required for detoxification of ethanol and aldehydes, is expanded (~10 copies vs. two in mouse and one in human). We also show that the beaver cells are more resistant to ethanol, and beaver liver extracts show higher ability to metabolize aldehydes than the mouse samples. Furthermore,Hpgd, a tumor suppressor gene, is uniquely duplicated in the beaver among rodents. Our evolutionary analysis identified beaver genes under positive selection which are associated with tumor suppression and longevity. Genes involved in lipid metabolism show positive selection signals, changes in copy number and altered gene expression in beavers. Several genes involved in DNA repair showed a higher expression in beavers which is consistent with the trend observed in other long-lived mammals. In summary, we identified several genes that likely contribute to beaver longevity and cancer resistance, including increased ability to detoxify aldehydes, enhanced tumor suppression and DNA repair, and altered lipid metabolism.
Multicompartmental Non-invasive Sensing of Postprandial Lipemia in Humans with Multispectral Optoacoustic Tomography
Authors: Fasoula Nikolina-Alexia , Karlas Angelos , Kallmayer Michael , Milik Anamaria Beatrice , Pelisek Jaroslav , Eckstein Hans-Henning , Klingenspor Martin , Ntziachristos Vasilis
Published: June 2020

Disturbed blood lipid profiles after food intake have been associated with increased risk for cardiovascular and metabolic disease. Postprandial lipid profiling (PLP) can be used as a risk factor for cardiovascular disease, insulin resistance and other metabolic diseases but is based today on frequent blood sampling over several hours after a meal, an approach that is invasive and inconvenient for patients. Non-invasive PLP may offer a favorable alternative for disseminated monitoring in humans. In this study, we investigate the use of localized lipid sensing guided by Multispectral Optoacoustic Tomography (MSOT) for non-invasive, label-free assessment of postprandial lipemia in human vasculature and in soft tissues. For penetrating deep in human tissue, we utilize measurements at 930 nm, where lipids exhibit strong light absorption in the near-infrared spectral range (NIR). In a pilot study, we longitudinally measured postprandial lipemia in healthy subjects over 6 hours following consumption of a high-fat meal. Localized measurements were obtained from four anatomical structures: the radial artery, the cephalic vein, the brachioradialis muscle and the subcutaneous fat of the forearm. Analysis of optoacoustic signals demonstrated a 63.4% mean lipid increase in intra-arterial lipids at approximately 4 hours postprandially, a 89.7% mean increase in intra-venous lipids at 3-hours, a 120.8% mean increase in intra-muscular lipids at 3-hours and a 30.5% mean increase in subcutaneous fat lipids at 4-hours. We discuss how portable MSOT offers unprecedented potential to study lipid metabolism that could lead to novel diagnostics and prevention strategies by offering label-free and non-invasive detection of tissue biomarkers implicated in cardiometabolic diseases.
Thin Film Metallization Stacks Serve as Reliable Conductors on Ceramic-based Substrates for Active Implants
Authors: Kiele Patrick , Čvančara Paul , Langenmair Michael , Mueller Matthias , Stieglitz Thomas
Published: June 2020

Hermetic and non-hermetic packages of active implantable medical devices are often fabricated of ceramics like alumina. Screen printed PtAu paste is the state of the art metallization for functional structures. Due to solid state and liquid diffusion of Au at thermal exposure, solder times are limited; otherwise metal structures tend to delaminate. Moreover, it was shown that PtAu with solder fails after 37.4 years. We established a thin film metallization on alumina process to overcome these disadvantages. The metallization consists of sputtered platinum with an underlying adhesion layer made of tungsten-titanium to increase the adhesion strength to the alumina substrate. We avoided using gold in this work due to its high diffusion tendency. Instead, the materials in use provide relatively low diffusion properties, which potentially increases the long term mechanical performance and usability during assembly and packaging.

Utilizing the Design of Experiment (DoE) methodology, we derived an optimal Pt thickness of 500 nm with 43 nm of WTi as adhesion promoting layer. After accelerated aging at 150 °C, corresponding to 125 years at body temperature (37 °C), the contact pad adhesion strength was with 32.75 MPa ± 7.08 MPa still significantly higher than the safety limit of 17 MPa, following the recommendations for a robust screen-printing metallization process. Moreover, soldering times of up to 120 s did not influence the adhesive strength. The new process reduced the minimum track distance to 50% of screen printing values and is capable of rapid prototyping. It helps to make the assembly process independent of the manufacturing person, to increase the yield of device fabrication and -most important in implantable device manufacturing-to make it more robust and thereby more safe for the patient.
Transgressive segregation for salt tolerance in rice due to physiological coupling and uncoupling and genetic network rewiring
Authors: Pabuayon Isaiah C.M. , Kitazumi Ai , Cushman Kevin R. , Singh Rakesh Kumar , Gregorio Glenn B. , Dhatt Balpreet , Zabet-Moghaddam Masoud , Walia Harkamal , de los Reyes Benildo G.
Published: June 2020

Transgressive segregation is common in plant breeding populations, where a small minority of recombinants are outliers relative to parental phenotypes. While this phenomenon has been attributed to complementation and epistatic effects, the physiological, biochemical, and molecular bases have not been fully illuminated. By systems-level scrutiny of the IR29 x Pokkali recombinant inbred population of rice, we addressed the hypothesis that novel salt tolerance phenotypes are created by positive or negative coupling or uncoupling effects and novel regulatory networks. Hyperspectral profiling distinguished the transgressive individuals in terms of stress penalty to growth. Non-parental network signatures that led to either optimal or non-optimal integration of developmental with stress-related mechanisms were evident at the macro-physiological, biochemical, metabolic, and transcriptomic levels. The large positive net gain in super-tolerant progeny was due to ideal complementation of beneficial traits, while shedding antagonistic traits. Super-sensitivity was explained by the stacking of multiple antagonistic traits and loss of major beneficial traits. The mechanisms elucidated in this study are consistent with the Omnigenic Theory, emphasizing the synergy or lack thereof between core and peripheral components. This study supports a breeding paradigm based on genomic modeling to create the novel adaptive phenotypes for the crops of the 21stcentury.
A survivin-driven tumour-activatable minicircle system for prostate cancer theranostics
Authors: Wang TianDuo , Chen Yuanxin , Goodale David , Allan Alison L. , Ronald John A.
Published: June 2020

Gene vectors driven by tumour-specific promoters to express reporter genes and therapeutic genes are an emerging approach for improved cancer diagnosis and treatment. Minicircles (MCs) are shortened plasmids stripped of prokaryotic sequences and have potency and safety characteristics that are beneficial for clinical translation. We previously developed survivin-driven, tumour-activatable MCs for cancer detection via a secreted blood reporter assay. Here we present a novel theranostic system for prostate cancer featuring a pair of survivin-driven MCs, combining selective detection of aggressive tumours via a urinary reporter test and subsequent tumour treatment with gene-directed enzyme prodrug therapy.

Methods

We engineered both diagnostic and therapeutic survivin-driven MCs expressing Gaussia luciferase, a secreted reporter that is detectable in the urine, and cytosine deaminase:uracil phosphoribosyltransferase fusion, respectively. Diagnostic MCs were evaluated in mice carrying orthotopic prostate tumours with varying survivin levels, measuring reporter activity in serial urine samples. Therapeutic MCs were evaluated in mice receiving prodrug using bioluminescence imaging to assess cancer cell viability over time.

Results

Diagnostic MCs revealed mice with aggressive prostate tumours exhibited significantly higher urine reporter activity than mice with non-aggressive tumours and tumour-free mice. Combined with 5-fluorocytosine prodrug treatment, therapeutic MCs resulted in reduced bioluminescence signal in mice with aggressive prostate tumours compared to control mice.

Conclusion

Sequential use of these MCs may be used to first identify patients carrying aggressive prostate cancer by a urinary reporter test, followed by stringent treatment in stratified individuals identified to have high-risk lesions. This work serves to highlight tumour-activatable MCs as a viable platform for development of gene-based tumour-activatable theranostics.
Heterogeneous adaptation of cysteine reactivity to a covalent oncometabolite
Authors: Perez Minervo , Bak Daniel W. , Bergholtz Sarah E. , Crooks Daniel R. , Yang Youfeng , Weerapana Eranthie , Linehan W. Marston , Meier Jordan L.
Published: June 2020

Metabolism and signaling intersect in the genetic cancer syndrome hereditary leiomyomatosis and renal cell carcinoma (HLRCC), a disease in which mutation of the TCA cycle enzyme fumarate hydratase (FH) causes hyperaccumulation of fumarate. This electrophilic oncometabolite can alter gene activity at the level of transcription, via reversible inhibition of epigenetic dioxygenases, as well as posttranslationally, via covalent modification of cysteine residues. To better understand how metabolites function as covalent signals, here we report a chemoproteomic analysis of a kidney-derived HLRCC cell line. Building on previous studies, we applied a general reactivity probe to compile a dataset of cysteine residues sensitive to rescue of cellular FH activity. This revealed a broad upregulation of cysteine reactivity upon FH rescue, caused by an approximately equal proportion of transcriptional and posttranslational regulation in the rescue cell line. Gene ontology analysis highlights new targets and pathways potentially modulated by FH mutation. Comparison of the new dataset to literature studies highlights considerable heterogeneity in the adaptive response of cysteine-containing proteins in different models of HLRCC. Our analysis provides a resource for understanding the proteomic adaptation to fumarate accumulation, and a foundation for future efforts to exploit this knowledge for cancer therapy.
Patient-derived xenograft culture-transplant system for investigation of human breast cancer metastasis
Authors: Ma Dennis , Hernandez Grace A. , Lefebvre Austin E.Y.T. , Alshetaiwi Hamad , Blake Kerrigan , Dave Kushal R. , Rauf Maha , Williams Justice W. , Davis Ryan T. , Evans Katrina T. , Masoud Madona Y.G. , Lee Regis , Edwards Robert A. , Digman Michelle A. , Kessenbrock Kai , Lawson Devon A.
Published: June 2020

Metastasis is a fatal disease where research progress has been hindered by a lack of authentic experimental models. Here, we develop a 3D tumor sphere culture-transplant system that facilitates the expansion and engineering of patient-derived xenograft (PDX) tumor cells for functional metastasis assaysin vivo. Orthotopic transplantation and RNA sequencing analyses show that PDX tumor spheres maintain tumorigenic potential, and the molecular marker and global transcriptome signatures of native tumor cells. Tumor spheres display robust capacity for lentiviral engineering and dissemination in spontaneous and experimental metastasis assaysin vivo. Inhibition of pathways previously reported to attenuate metastasis also inhibit metastasis after sphere culture, validating our approach for authentic investigations of metastasis. Finally, we demonstrate a new role for the metabolic enzyme NME1 in promoting breast cancer metastasis, providing proof-of-principle that our culture-transplant system can be used for authentic propagation and engineering of patient tumor cells for functional studies of metastasis.
Virus-Receptor Interactions of Glycosylated SARS-CoV-2 Spike and Human ACE2 Receptor
Authors: Zhao Peng , Praissman Jeremy L. , Grant Oliver C. , Cai Yongfei , Xiao Tianshu , Rosenbalm Katelyn E. , Aoki Kazuhiro , Kellman Benjamin P. , Bridger Robert , Barouch Dan H. , Brindley Melinda A. , Lewis Nathan E. , Tiemeyer Michael , Chen Bing , Woods Robert J. , Wells Lance
Published: June 2020

The current COVID-19 pandemic is caused by the SARS-CoV-2 betacoronavirus, which utilizes its highly glycosylated trimeric Spike protein to bind to the cell surface receptor ACE2 glycoprotein and facilitate host cell entry. We utilized glycomics-informed glycoproteomics to characterize site-specific microheterogeneity of glycosylation for a recombinant trimer Spike mimetic immunogen and for a soluble version of human ACE2. We combined this information with bioinformatic analyses of natural variants and with existing 3D-structures of both glycoproteins to generate molecular dynamics simulations of each glycoprotein alone and interacting with one another. Our results highlight roles for glycans in sterically masking polypeptide epitopes and directly modulating Spike-ACE2 interactions. Furthermore, our results illustrate the impact of viral evolution and divergence on Spike glycosylation, as well as the influence of natural variants on ACE2 receptor glycosylation that, taken together, can facilitate immunogen design to achieve antibody neutralization and inform therapeutic strategies to inhibit viral infection.
Megakaryopoiesis in Dengue virus infected K562 cell promotes viral replication which inhibits endomitosis and accumulation of ROS associated with differentiation
Authors: Kaur Jaskaran , Rawat Yogita , Sood Vikas , Rathore Deepak K. , Kumar Shrikant K. , Kumar Niraj K. , Bhattacharyya Sankar
Published: June 2020

In the human host blood Monocytes and bone marrow Megakaryocytes are implicated as major sites supporting high replication. The human K562 cell line supports DENV replication and represent Megakaryocyte-Erythrocyte progenitors (MEP), replicating features ofin vivoMegakaryopoiesis upon stimulation with Phorbol esters. In this article, we report results that indicate the mutual influence of Megakaryopoiesis and DENV replication on each other, through comparison of PMA-induced differentiation of either mock-infected or DENV-infected K562 cells. We present data showing PMA-induced differentiation to drastically increase DENV replication and a concomitant augmented secretion of infectious virus. Although the mechanism is not clear yet, we show that it is not through an increased uptake of virus by differentiated cells. On the other hand, DENV replication in cells undergoing PMA-induced differentiation, interferes with major differentiation markers of Megakaryopoiesis including activation of ERK1/2 MAP Kinase, endomitosis and surface expression of platelet-specific proteins without any drastic effect on cell death. Among signaling intermediaries of the JAK-STAT pathway, we observed infection associated degradation of SOC3 protein similar to earlier observations with STAT2. DENV infection leads to accumulation of Reactive-oxygen species (ROS) in different cells including K562. PMA-induced differentiation of uninfected K562 cells also leads to intracellular ROS accumulation. Interestingly, we observed ROS accumulation to be suppressed by concomitant DENV replication in K562 cells undergoing PMA-induced differentiation. This is the first report of a model system where DENV replication suppresses intracellular ROS accumulation. The implications of these results for Megakaryopoiesis and viral replication would be discussed.
In-vitro study of the activity of some medicinal plant leaf extracts on urinary tract infection causing bacterial pathogens isolated from indigenous people of Bolangir district, Odisha, India
Authors: Saha Mirkashim H. , Dash Saumya , Mishra Abhisek , Satpathy Jyotismita , Mishra Sujit K.
Published: June 2020

Thousands of people over the world are suffering from Urinary tract infection (UTI) each day. Side effects associated with the most preferred allopathic treatment for UTI and antibiotic resistance shown by the pathogens has evolved as major challenge. Owing to this, the present study was carried out to identify the bacterial UTI pathogens among indigenous people of Bolangir district, Odisha, Indiavis-à-visexplore the alternative source of natural therapeutics. Bacterial pathogens causing the UTIs were identified using the colony morphology, Gram’s staining and biochemical characterization techniques. Microbial inhibitory test of antibiotics and leaf extracts were performed using the disc diffusion and well diffusion techniques respectively. MIC and MLC were determined by broth dilution method. Binary probit model was used to determine the prevalence of UTI across people of different age group and gender. Four bacterial strains namelyEscherichia coli,Enterococcus faecalis,Staphylococcus aureus, andKlebsiella pneumoniaewere identified as the causative agents of UTI among the people. TheE. coliwas identified as most infectious whileS. aureusas the least infectious pathogen. Females in the age group of ‘16-30 years’ and male within ‘61-75 years’ were more susceptible to UTI. Among the tested leaf extracts,Tamarindus indicaandClitoria ternateawere more effective to treat UTI as compared to the tested antibiotics Ceftriaxone and Piperacillin. The leaf extracts ofT. indicaandC. ternateamay effectively be utilized for herbal drug development for treatment of UTI.
In Vitroefficacy comparison of linezolid, tedizolid, sutezolid and delpazolid against rapid growing Mycobacteria isolated in Beijing, China
Authors: Wen Shuan , Gao Xiaopan , Zhao Weijie , Huo Fengmin , Jiang Guanglu , Dong Lingling , Zhao Liping , Wang Fen , Yu Xia , Huang Hairong
Published: June 2020

The natural resistance of rapid growth Mycobacterium (RGM) against multiple antibiotics renders the treatment of caused infections less successful and time consuming. Therefore, new effective agents are urgently needed. The aim of this study was to evaluate thein vitrosusceptibility of 115 isolates, constituting different RGM species, against four oxazolidinones i.e. delpazolid, sutezolid, tedizolid and linezolid. Additionally, 32 reference strains of different RGM species were also tested. The four oxazolidinones exhibited potentin vitroactivity against the recruited RGM reference strains, 24 out of 32 RGM species had MICs ≤ 8 μg/mL against all four oxazolidinones whereas tedizolid and delpazolid generally presented lower MICs than linezolid or sutezolid. Tedizolid showed the strongest activity against clinical isolates ofM. abscessuswith MIC50=1μg/mL and MIC90=2μg/mL. MIC values for tedizolid were usually 4- to 8-fold less than the MICs of linezolid forM. abscessussubsp.abscessus. The MIC distributions of sutezolid and linezolid were similar, while delpazolid showed 2-fold lower MIC as compared with linezolid. Linezolid was not active against most of the testedM. fortuitumisolates, 22 out of the 25M. fortuitumwere resistant against linezolid. However, delpazolid exhibited better antimicrobial activity against these isolates with 4-fold lower MIC values, in contrast with linezolid. In addition, the protein alignment of RplC and RplD and structure based analysis showed that there may be no correlation between oxazolidinones resistance and mutationsin rplC, rplDand23 srRNAgenes in tested RGM. This study showed tedizolid harbors the strongest inhibitory activity againstM. abscessus in vitro, while delpazolid presented the best activity againstM. fortuitum, which provided important insights on the potential clinical application of oxazolidinones to treat RGM infections.
A link between zinc uptake, bile salts, and a capsule required for virulence of a mastitis-associated extraintestinal pathogenicEscherichia colistrain
Authors: Olson Michael A. , Grimsrud Aleksander , Richards Amanda C. , Mulvey Matthew A. , Wilson Eric , Erickson David L.
Published: June 2020

Extraintestinal pathogenicEscherichia coli(ExPEC) are major causes of urinary and bloodstream infections. ExPEC reservoirs are not completely understood. Some mastitis-associatedE. coli(MAEC) strains carry genes associated with ExPEC virulence, including metal scavenging, immune avoidance, and host attachment functions. In this study, we investigated the role of the high-affinity zinc uptake (znuABC) system in the MAEC strain M12. Elimination ofznuABCmoderately decreased fitness during mouse mammary gland infections. The ΔznuABCmutant strain exhibited an unexpected growth delay in the presence of bile salts, which was alleviated by the addition of excess zinc. We isolated ΔznuABCmutant suppressor mutants with improved growth of in bile salts, several of which no longer produced the K96 capsule made by strain M12. Addition of bile salts also reduced capsule production by strain M12 and ExPEC strain CP9, suggesting that capsule synthesis may be detrimental when bile salts are present. To better understand the role of the capsule, we compared the virulence of mastitis strain M12 with its unencapsulated ΔkpsCSmutant in two models of ExPEC disease. The wild type strain successfully colonized mouse bladders and kidneys and was highly virulent in intraperitoneal infections. Conversely, the ΔkpsCSmutant was unable to colonize kidneys and was unable to cause sepsis. These results demonstrate that some MAEC may be capable of causing human ExPEC illness. Virulence of strain M12 in these infections is dependent on its capsule. However, capsule may interfere with zinc homeostasis in the presence of bile salts while in the digestive tract.