bluuebunny/crossref_metadata_2025 huggingface dataset:
Cyclic Loading Induces Anabolic and Catabolic Gene Expression in ACLs in a Load-Dependent and Sex-Specific Manner
Authors: Paschall Lauren , Carrozzi Sabrina , Tabdanov Erdem , Dhawan Aman , Szczesny Spencer
Published: November 2022

Anterior cruciate ligament (ACL) injuries are historically thought to be a result of a single acute overload or traumatic event. However, recent studies suggest that ACL failure may be a consequence of fatigue damage. Additionally, the remodeling response of ACLs to fatigue loading is unknown. Therefore, the objective of this study was to investigate the remodeling response of ACLs to cyclic loading. Furthermore, given that women have an increased rate of ACL rupture, we investigated whether this remodeling response is sex specific. ACLs were harvested from male and female New Zealand white rabbits and cyclically loaded in a tensile bioreactor mimicking the full range of physiological loading (2, 4, and 8 MPa). Expression of markers for anabolic and catabolic tissue remodeling, as well as inflammatory cytokines, was quantified using RT-qPCR. We found that the expression of markers for tissue remodeling of the ACL is dependent on the magnitude of loading and is sex specific. Male ACLs activated a tissue remodeling response to cyclic loading below 4 MPa loads but turned off remodeling at 8 MPa. These data support the hypothesis that noncontact ACL injury is a consequence of failed tissue remodeling and inadequate repair of microtrauma resulting from fatigue loading. Conversely, female ACLs downregulate genes responsible for tissue remodeling in response to cyclic loading at all magnitudes, which may explain the increased rate of ACL tears in women. Together, these data provide insight into the remodeling response of ACLs in vivo and potentially offer novel approaches for preventing ACL rupture.
Glucocerebrosidase activity and lipid levels are related to protein pathologies in Parkinson’s disease
Authors: Leyns Cheryl E. G. , Prigent Alice , Beezhold Brenna , Yao Lihang , Hatcher Nathan , Tao Peining , Kang John , Suh EunRan , Van Deerlin Vivianna M , Trojanowski John Q. , Lee Virginia M.Y. , Kennedy Matthew , Fell Matthew , Henderson Michael X.
Published: November 2022

Parkinson’s disease (PD) and dementia with Lewy bodies (DLB) are progressive neurodegenerative diseases characterized by the accumulation of misfolded α-synuclein in the form of Lewy pathology. While most cases are sporadic, there are rare genetic mutations that cause disease and more common variants that increase incidence of disease. The most prominent genetic mutations for PD and DLB are in theGBA1andLRRK2genes.GBA1mutations are associated with decreased glucocerebrosidase activity and lysosomal accumulation of its lipid substrates, glucosylceramide and glucosylsphingosine. Previous studies have shown a link between this enzyme and lipids even in sporadic PD. However, it is unclear how the protein pathologies of disease are related to enzyme activity and glycosphingolipid levels. To address this gap in knowledge, we examined quantitative protein pathology, glucocerebrosidase activity and lipid substrates in parallel from 4 regions of 91 brains with no neurological disease, idiopathic,GBA1-linked, orLRRK2-linked PD and DLB. We find that several biomarkers are altered with respect to mutation and disease severity. We found mild association of glucocerebrosidase activity with disease, but a strong association of glucosylsphingosine with α-synuclein pathology, irrespective of genetic mutation. This association suggests that Lewy pathology precipitates changes in lipid levels related to disease severity.
Emergent Electrical Activity, Tissue Heterogeneity, and Robustness in a Calcium Feedback Regulatory Model of the Sinoatrial Node
Authors: Moise Nicolae , Weinberg Seth H.
Published: November 2022

The sinoatrial node (SAN) is the primary pacemaker of the heart. SAN activity emerges at an early point in life and maintains a steady rhythm for the lifetime of the organism. The ion channel composition and currents of SAN cells can be influenced by a variety of factors. Therefore, the emergent activity and long-term stability imply some form of dynamical feedback control of SAN activity. We adapt a recent feedback model - previously utilized to describedion conductances in neurons - to a model of SAN cells and tissue. The model describes a minimal regulatory mechanism of ion channel conductances via feedback between intracellular calcium and an intrinsic target calcium level. By coupling a SAN cell to the calcium feedback model, we show that spontaneous electrical activity emerges from quiescence and is maintained at steady-state. In a 2D SAN tissue, spatial variability in intracellular calcium targets lead to significant, self-organized heterogeneous ion channel expression and calcium transients throughout the tissue. Further, multiple pacemaking regions appear, which interact and lead to time-varying cycle length, demonstrating that variability in heart rate is an emergent property of the feedback model. Finally, we demonstrate that the SAN tissue is robust to the silencing of leading cells or ion channel knockouts. Thus, the calcium feedback model can reproduce and explain many fundamental emergent properties of activity in the SAN that have been observed experimentally based on a minimal description of intracellular calcium and ion channel regulatory networks.

Key Points Summary

The robust function of the sinoatrial node (SAN) is reproduced in an intracellular calcium feedback model governing ion channel conductances

The feedback model predicts the emergence and long-term maintenance of spontaneous oscillatory electrical activity

Integrating the feedback model into a 2D SAN tissue leads to emergent spatial heterogeneity, multiple pacemaking regions, and variable cycle length

SAN cells and tissue with feedback are robust to cell injury and channel knock-outs
Brain-wide representations of behavior spanning multiple timescales and states inC. elegans
Authors: Atanas Adam A. , Kim Jungsoo , Wang Ziyu , Bueno Eric , Becker McCoy , Kang Di , Park Jungyeon , Estrem Cassi , Kramer Talya S. , Baskoylu Saba , Mansingkha Vikash K. , Flavell Steven W.
Published: November 2022

Changes in an animal’s behavior and internal state are accompanied by widespread changes in activity across its brain. However, how neurons across the brain encode behavior and how this is impacted by state is poorly understood. We recorded brain-wide activity and the diverse motor programs of freely-movingC. elegansand built probabilistic models that explain how each neuron encodes quantitative features of the animal’s behavior. By determining the identities of the recorded neurons, we created, for the first time, an atlas of how the defined neuron classes in theC. elegansconnectome encode behavior. Many neuron classes have conjunctive representations of multiple behaviors. Moreover, while many neurons encode current motor actions, others encode recent actions. Changes in behavioral state are accompanied by widespread changes in how neurons encode behavior, and we identify these flexible nodes in the connectome. Our results provide a global map of how the cell types across an animal’s brain encode its behavior.
3D-bioprinted, phototunable hydrogel models for studying adventitial fibroblast activation in pulmonary arterial hypertension
Authors: Davis-Hall Duncan , Thomas Emily , Peña Brisa , Magin Chelsea M.
Published: November 2022

Pulmonary arterial hypertension (PAH) is a progressive disease of the lung vasculature, characterized by elevated pulmonary blood pressure, remodeling of the pulmonary arteries, and ultimately right ventricular failure. Therapeutic interventions for PAH are limited in part by the lack ofin vitroscreening platforms that accurately reproduce dynamic arterial wall mechanical properties. Here we present a 3D-bioprinted model of the pulmonary arterial adventitia comprised of a phototunable poly(ethylene glycol) alpha methacrylate (PEG-αMA)-based hydrogel and primary human pulmonary artery adventitia fibroblasts (HPAAFs). This unique biomaterial emulates PAH pathogenesisin vitrothrough a two-step polymerization reaction. First, PEG-αMA macromer was crosslinked off-stoichiometry by 3D bioprinting an acidic bioink solution into a basic gelatin support bath initiating a base-catalyzed thiol-ene reaction with synthetic and biodegradable crosslinkers. Then, matrix stiffening was induced by photoinitiated homopolymerization of unreacted αMA end groups. A design of experiments approach produced a hydrogel platform that exhibited an initial elastic modulus (E) within the range of healthy pulmonary arterial tissue (E = 4.7 ± 0.09 kPa) that was stiffened to the pathologic range of hypertensive tissue (E = 12.8 ± 0.47 kPa) and supported cellular proliferation over time. A higher percentage of HPAAFs cultured in stiffened hydrogels expressed the fibrotic marker alpha-smooth muscle actin than cells in soft hydrogels (88 ± 2% versus 65 ± 4%). Likewise, a greater percentage of HPAAFs were positive for the proliferation marker 5-ethynyl-2’-deoxyuridine (EdU) in stiffened models (66 ± 6%) compared to soft (39 ± 6%). These results demonstrate that 3D-bioprinted, phototunable models of pulmonary artery adventitia are a tool that enable investigation of fibrotic pathogenesisin vitro.
SUMO2 Protects Against Tau-induced Synaptic and Cognitive Dysfunction
Authors: Orsini Franca , Argyrousi Elentina , Restelli Elena , Ford Lenzie K. , Takamura Hironori , Matsuzaki Shinsuke , Zentilin Lorena , Pascente Rosaria , Kanaan Nicholas M , Soni Rajesh , Katayama Taiichi , Chiesa Roberto , Forloni Gianluigi , Kosik Kenneth S. , Kandel Eric R. , Fraser Paul E. , Arancio Ottavio , Fioriti Luana
Published: November 2022

Abnormal intracellular accumulation of Tau aggregates is a hallmark of Alzheimer’s disease (AD) and other Tauopathies, such as Frontotemporal dementia (FTD), which can be caused by mutations of Tau. Mutated and pathological Tau can undergo a range of post-translational modifications (PTMs) that might trigger or modulate disease pathology. Recent studies indicate that modification of wild type Tau bySmallubiquitin-likemodifier SUMO isoform 1 (SUMO1) controls Tau hyperphosphorylation and aggregation, suggesting that SUMOylation acts as a central regulator of Tau’s biochemical properties. Besides SUMO1, Tau is modified by SUMO2/3, however the consequences of this modification have not been investigated. Here, using viral approaches on primary hippocampal neurons, transgenic mice expressing mutant Tau and SUMO2, and iPSC-derived neurons from FTD patients, we evaluated whether SUMO2/3 conjugation modifies the neurodegenerative disease pathology associated with the aggregation-prone mutant Tau P301L, P301S, and R406W variants. We found that mutant forms of Tau are targets of SUMO2/3, and SUMO2/3 conjugation is neuroprotective. Importantly, expression of mutant Tau is accompanied by a significant reduction of SUMO2/3 conjugation levels, and restoring levels of SUMO2 reduces mutant Tau aggregation and phosphorylation in all model systems Furthermore, overexpression of SUMO2 restores levels of pre- and post-synaptic markers, associated with a complete rescue of the LTP and memory deficits in transgenic mice expressing mutant Tau. These findings bring to light the potential therapeutic implication of manipulating SUMO conjugation to detoxify Tau through PTM-based approaches.
Hemodynamic timing in resting-state and breathing-task BOLD fMRI
Authors: Gong Jingxuan , Stickland Rachael C. , Bright Molly G.
Published: November 2022

The blood flow response to a vasoactive stimulus demonstrates regional heterogeneity across both the healthy brain and in cerebrovascular pathology. The timing of a regional hemodynamic response is emerging as an important biomarker of cerebrovascular dysfunction, as well as a confound within fMRI analyses. Previous research demonstrated that hemodynamic timing is more robustly characterized when a larger systemic vascular response is evoked by a breathing challenge, compared to when only spontaneous fluctuations in vascular physiology are present (i.e., in resting-state data). However, it is not clear whether hemodynamic delays in these two conditions are physiologically interchangeable, and how methodological signal-to-noise factors may limit their agreement. To address this, we generated whole-brain maps of hemodynamic delays in nine healthy adults. We assessed the agreement of voxel-wise gray matter (GM) hemodynamic delays between two conditions: resting-state and breath-holding. We found that delay values demonstrated poor agreement when considering all GM voxels, but increasingly greater agreement when limiting analyses to voxels showing strong correlation with the GM mean time-series. Voxels showing the strongest agreement with the GM mean time-series were primarily located near large venous vessels, however these voxels explain some, but not all, of the observed agreement in timing. Increasing the degree of spatial smoothing of the fMRI data enhanced the correlation between individual voxel time-series and the GM mean time-series. These results suggest that signal-to-noise factors may be limiting the accuracy of voxel-wise timing estimates and hence their agreement between the two data segments. In conclusion, caution must be taken when using voxel-wise delay estimates from resting-state and breathing-task data interchangeably, and additional work is needed to evaluate their relative sensitivity and specificity to aspects of vascular physiology and pathology.
Differential modulation of cellular phenotype and drug sensitivity by extracellular matrix proteins in primary and metastatic pancreatic cancer cells
Authors: Usman Olalekan H , Kumar Sampath , Walker Reddick R , Xie Gengqiang , Sumajit Hyeje , Jalil AbdelAziz R. , Ramakrishnan Subramanian , Dooling Lawrence J , Wang Yue Julia , Irianto Jerome
Published: November 2022

Pancreatic cancer adenocarcinoma (PDAC) is reported to be the third highest cause of cancer-related deaths in the United States. PDAC is known for its high proportion of stroma, which accounts for 90% of the tumor mass. The stroma is made up of extracellular matrix (ECM) and non-malignant cells such as inflammatory cells, cancer-associated fibroblasts, and lymphatic and blood vessels. Here, we decoupled the effects of the ECM on PDAC cell lines by culturing cells on surfaces coated with different ECM proteins. Our data show that the primary tumor-derived cell lines have different morphology depending on the ECM proteins on which they are cultured, while metastatic lesion-derived PDAC lines’ morphology does not change with respect to the different ECM proteins. Similarly, ECM proteins modulate the proliferation rate and the gemcitabine sensitivity of the primary tumor PDAC cell lines, but not the metastatic PDAC lines. Lastly, transcriptomics analysis of the primary tumor PDAC cells cultured on different ECM proteins reveals the regulation of various pathways, such as cell cycle, cell adhesion molecules, and focal adhesion, including the regulation of several integrin genes that are essential for ECM recognition.
Characterizing microplastic ingestion, transformation, and excretion in insects using fluorescent plastics
Authors: Ritchie Marshall W. , Cheslock Alexandra , Bourdages Madelaine P.T. , Hamilton Bonnie M. , Provencher Jennifer F. , Allison Jane E. , MacMillan Heath A.
Published: November 2022

Plastic pollution is a growing threat to our natural environment. Plastic waste/pollution results from high emissions of both macro (> 5 mm) and microplastics (MPs; < 5 mm) as well as environmental fractioning of macroplastics into microplastics. Microplastics have been shown to have a range of negative impacts on biota. Harmonized methods to accurately measure and count MPs from animal samples are limited, but what methods exist are not ideal for a controlled laboratory environment where plastic ingestion, transformation, and elimination can be quantified and related to molecular, physiological, and organismal traits. Here we propose a complete method for isolating and characterizing fluorescent MPs by combining several previously reported approaches into one comprehensive workflow. We combine tissue dissection, organic material digestion, sample filtering, and automated imaging techniques to show how fluorescently-labelled MPs provided to animals (e.g. in their diet) in a laboratory setting can be isolated, identified, and quantified. As a proof of concept, we fed crickets (Gryllodes sigillatus) a diet of 2.5% (w/w) fluorescently-labelled plastics and isolated and characterized plastic particles within the gut and frass.
Phenotypic, genomic, and transcriptomic heterogeneity in a pancreatic cancer cell line
Authors: Xie Gengqiang , Zhang Liting , Usman Olalekan H , Kumar Sampath , Modak Chaity , Patel Dhenu , Kavanaugh Megan , Mallory Xian , Wang Yue Julia , Irianto Jerome
Published: November 2022

Objectives

To evaluate the suitability of the MIA PaCa-2 cell line for studying pancreatic cancer intratumor heterogeneity, we aim to further characterize the nature of MIA PaCa-2 cells’ phenotypic, genomic, and transcriptomic heterogeneity.

Methods

MIA PaCa-2 single-cell clones were established through flow cytometry. For the phenotypic study, we quantified the cellular morphology, proliferation rate, migration potential, and drug sensitivity of the clones. The chromosome copy number and transcriptomic profiles were quantified using SNPa and RNA-seq, respectively.

Results

Four MIA PaCa-2 clones showed distinctive phenotypes, with differences in cellular morphology, proliferation rate, migration potential, and drug sensitivity. We also observed a degree of genomic variations between these clones in form of chromosome copy number alterations and single nucleotide variations, suggesting the genomic heterogeneity of the population, and the intrinsic genomic instability of MIA PaCa-2 cells. Lastly, transcriptomic analysis of the clones also revealed gene expression profile differences between the clones, including the uniquely regulatedITGAV, which dictates the morphology of MIA PaCa-2 clones.

Conclusions

MIA PaCa-2 is comprised of cells with distinctive phenotypes, heterogeneous genomes, and differential transcriptomic profiles, suggesting its suitability as a model to study the underlying mechanisms behind pancreatic cancer heterogeneity.
Individual regional associations between Aβ-, tau- and neurodegeneration (ATN) with microglial activation in patients with primary and secondary tauopathies
Authors: Finze Anika , Biechele Gloria , Rauchmann Boris-Stephan , Franzmeier Nicolai , Palleis Carla , Katzdobler Sabrina , Weidinger Endy , Guersel Selim , Schuster Sebastian , Harris Stefanie , Schmitt Julia , Beyer Leonie , Gnörich Johannes , Lindner Simon , Albert Nathalie L. , Wetzel Christian , Rupprecht Rainer , Rominger Axel , Danek Adrian , Burow Lena , Kurz Carolin , Tato Maia , Utecht Julia , Papazov Boris , Zaganjori Mirlind , Trappmann Lena-Katharina , Goldhardt Oliver , Grimmer Timo , Haeckert Jan , Janowitz Daniel , Buerger Katharina , Keeser Daniel , Stoecklein Sophia , Dietrich Olaf , Morenas-Rodriguez Estrella , Barthel Henryk , Sabri Osama , Bartenstein Peter , Simons Mikael , Haass Christian , Höglinger Günter U. , Levin Johannes , Perneczky Robert , Brendel Matthias
Published: November 2022

β-amyloid (Aβ) and tau aggregation as well as neuronal injury and atrophy (ATN) are the major hallmarks of Alzheimer’s disease (AD), and biomarkers for these hallmarks have been linked to neuroinflammation. However, the detailed regional associations of these biomarkers with microglial activation in individual patients remain to be elucidated.

We investigated a cohort of 55 patients with AD and primary tauopathies and 10 healthy controls that underwent TSPO-, A-, tau-, and perfusion-surrogate-PET, as well as structural MRI. Z-score deviations for 246 brain regions were calculated and biomarker contributions of Aβ (A), tau (T), perfusion (N1) and gray matter atrophy (N2) to microglial activation (TSPO, I) were calculated for each individual subject. Individual ATN-related microglial activation was correlated with clinical performance and CSF soluble TREM2 (sTREM2) concentrations.

In typical and atypical AD, regional tau was stronger and more frequently associated with microglial activation when compared to regional A (AD: βT= 0.412±0.196 vs. βA= 0.142±0.123, p < 0.001; AD-CBS: βT= 0.385±0.176 vs. βA= 0.131±0.186, p = 0.031). The strong association between regional tau and microglia reproduced well in primary tauopathies (βT= 0.418±0.154). Stronger individual associations between tau and microglial activation were associated with poorer clinical performance. In patients with 4RT, sTREM2 levels showed a positive association with tau-related microglial activation.

Tau pathology has strong regional associations with microglial activation in primary and secondary tauopathies. An index of tau- and Aβ-associated microglia activation accounts for regional heterogeneity and allows for clinical and biomarker correlations with ATN-specific neuroinflammation.
Clinical profiles and risk factors of hepatocellular carcinoma with viral hepatitis at a tertiary hospital in Mogadishu
Authors: Hassan Mohammed A , Bashir Ahmed M , Akuku Isaiah G , Evelia Janet , Aweis Ahmed
Published: November 2022

Objectives

To describe the clinical profiles of HCC patients with or without viral hepatitis. Additionally, to evaluate the risk factors for HCC with viral hepatitis.

Methods

In a cross-sectional study, 52 HCC patients diagnosed by imaging in Mogadishu were enrolled. Serum liver blood samples were taken, and sociodemographic and risk data recorded. In addition, we used logistic regression to evaluate relevant sociodemographic and clinical risk factors for HCC with viral hepatitis.

Results

Of all patients, 86.5% were men, a median age of 64.0 years (interquartile range [IQR]: 51.5 to 72.0). 50% of the patients were cases of HBV or HCV, 44.2% was HBV and 15.4% were HCV. Diabetes, fatty liver disease and underweight was present in 23.1%, 30.8%, and 32.7%, respectively. The median of Gamma-glutamyl transferase (GGT) was 176.0 U/L (IQR 106.0–302.2 U/L; normal 1-60 U/L) and alpha-fetoprotein was 1337.5 ng/ml (IQR 566–2625; normal 0-8.78 ng/ml). Aspartate aminotransferase (AST) (121.5; IQR 64.5–198.0; normal range 0-31U/L), alkaline phosphatase (ALP) (173.5; IQR 125.2–232.5; normal 30-155 U/L) and direct bilirubin 0.5 (0.2–0.9; normal 0.01-0.4mg/dl) were also elevated. AST (median greatest dimension 164.5 U/L versus 70.0 U/L; p<0.001) and ALT (55.0 versus 30.0; p=0.048), and the distribution of international normalized ratio (INR) was higher (69.2% versus 34.6%; p=0.025) in viral hepatitis-positive patients than the non-viral group. Adjusted analysis showed INR was associated with viral hepatitis-HCC.

Conclusion

Most HCC patients were old and presented with HBV infection. Age, AST, ALT and INR biomarkers appeared to be influencing factors of viral hepatitis with HCC. The coexistence of viral hepatitis and metabolic factors enhanced the HCC development. In the primary care setting, evaluating the clinical profile could support risk stratification for HCC patients, including viral hepatitis-positive patients, timely detection, and decision-making in patient management.
Excess Mortality and Years of Potential Life Lost Among Black People in the US from 1999 to 2020
Authors: Caraballo César , Massey Daisy S. , Ndumele Chima D. , Haywood Trent , Kaleem Shayaan , King Terris , Liu Yuntian , Lu Yuan , Nunez-Smith Marcella , Taylor Herman A. , Watson Karol E. , Herrin Jeph , Yancy Clyde W. , Faust Jeremy S. , Krumholz Harlan M.
Published: November 2022

Importance

Amid efforts in the United States to promote health equity, there is a need to assess progress in reducing excess deaths and years of potential life lost (YPLL) among Black people compared with White people.

Objective

To evaluate trends in excess mortality and YPLL among Black people compared with White people.

Design

Serial cross-sectional.

Setting

National data from the Centers for Disease Control and Prevention, 1999-2020

Participants

Non-Hispanic White and non-Hispanic Black people

Exposures

Race as documented in the death certificates.

Main outcomes and measures

Excess age-adjusted all-cause and disease-specific mortality rate (per 100,000 individuals) and YPLL among Black people compared with White people.

Results

From 1999 to 2020, the total number of excess deaths was 658,356 and 1,154,108 among Black females and males, representing 34,938,070 and 47,005,048 excess YPLL among Black females and males. The excess deaths and YPLL were largest among infants and non-elderly adults. Heart disease had the most excess deaths. From 1999, the age-adjusted excess mortality rate declined at an annual average of -9.0 (95% CI: -10.0, -8.0; P<0.001) until 2015 among Black women and at an annual average of -16.3 (95% CI: -20.9, -11.6; P<0.001) until 2012 among Black men, followed by no significant change until 2019 in either group. From 2019 to 2020, excess deaths increased from 90.4 to 192 per 100,000 Black women and from 209.8 to 395 per 100,000 Black men, reaching rates approximating those of 1999. The trends in rates of excess YPLL followed a similar pattern.

Conclusions and relevance

Over a recent 22-year period, Black people in the US lost more than 80 million years of life when compared with White people. After a period of progress, improvements stalled, and most gains were eliminated in 2020.

KEY POINTS

Question

How many excess deaths and years of potential life lost (YPLL) for Black people, compared with White people, occurred in the United States from 1999 through 2020?

Findings

Based on Centers for Disease Control and Prevention data, excess deaths and YPLL persisted throughout the period, with initial progress followed by little improvement, and then worsening in 2020 to about 1999 levels. Black people had 1.8 million excess deaths and over 80 million YPLL over the study period.

Meaning

After initial progress, excess mortality and YPLL among Black people stagnated and then worsened, indicating a need for new approaches.
Evaluation and Clinical Validation of Monkeypox Virus Real-Time PCR Assays
Authors: Mills Margaret G. , Juergens Kate B. , Gov Jolene , McCormick Carter , Sampoleo Reigran , Kachikis Alisa , Amory John K. , Fang Ferric C. , Pérez-Osorio Ailyn C. , Lieberman Nicole A. P. , Greninger Alexander L.
Published: November 2022

BACKGROUND

In spring of 2022, an outbreak of monkeypox spread worldwide. Here, we describe performance characteristics of monkeypox virus (MPXV)-specific and pan-orthopoxvirus qPCR assays for clinical use.

METHODS

We validated probe-based qPCR assays targeting MPXV-specific loci F3L and G2R (genes MPXVgp052/OPG065 and MPXVgp002 and gp190/OPG002, respectively) and a pan-orthopoxvirus assay targeting the E9L locus (MPXVgp057/OPG071). Clinical samples and synthetic controls were extracted using the Roche MP96 or Promega Maxwell 48 instrument. qPCR was performed on the AB7500 thermocycler. Synthetic control DNA and high concentration clinical samples were quantified by droplet PCR. Cross-reactivity was evaluated for camelpox and cowpox genomic DNA, vaccinia culture supernatant, and HSV- and VZV-positive clinical specimens. We also determined analytical performance of the F3L assay using dry swabs, Aptima vaginal and rectal swabs, nasopharyngeal, rectal, and oral swabs, cerebrospinal fluid, plasma, serum, whole blood, breastmilk, urine, saliva, and semen.

RESULTS

The MPXV-F3L assay is reproducible at a limit of detection (LoD) of 65.6 copies/mL of viral DNA in VTM/UTM, or 3.3 copies/PCR reaction. No cross-reactivity with herpesviruses or other poxviruses was observed. MPXV-F3L detects MPXV DNA in alternative specimen types, with an LoD ranging between 260-1000 copies/mL, or 5.7-10 copies/PCR reaction. In clinical swab VTM specimens, MPXV-F3L and MPXV-G2R assays outperformed OPXV-E9L by an average of 2.4 and 2.8 Cts, respectively. MPXV-G2R outperformed MPXV-F3L by 0.4 Cts, consistent with presence of two copies of G2R present in labile inverted terminal repeats (ITRs) of MPXV genome.

CONCLUSIONS

MPXV is readily detected by qPCR using three clinically validated assays.
A Delphi Process for Reaching Nationwide Consensus on Antimicrobial Intravenous-to-oral Switch Criteria and Development of an Antimicrobial Intravenous-to-oral Switch Decision Aid
Authors: Harvey Eleanor J , Hand Kieran , Weston Dale , Ashiru-Oredope Diane
Published: November 2022

Introduction

Antimicrobial stewardship (AMS) strategies, such as intravenous-to-oral switch (IVOS), promote optimal antimicrobial use, contributing to safer and more effective patient care and tackling antimicrobial resistance (AMR).

Aim

This study aimed to achieve nationwide multidisciplinary expert consensus on antimicrobial IVOS criteria for timely switch in hospitalised adult patients and to design an IVOS decision aid to operationalise agreed IVOS criteria in the hospital setting.

Method

A 4-step Delphi process was chosen to achieve expert consensus on IVOS criteria and decision aid; it included Step One) Pilot/1stround questionnaire, Step Two) Virtual meeting, Step Three) 2ndround questionnaire and Step 4) Workshop. This study follows the Appraisal of Guidelines for Research and Evaluation II instrument checklist.

Results

Step One questionnaire of 42 IVOS criteria had 24 respondents, 15 of whom participated in Step Two, where 37 criteria were accepted for the next step. Step Three had 242 respondents (England n=195, Northern Ireland n=18, Scotland n=18, Wales n=11); 27 criteria were accepted. Step Four had 48 survey respondents and 33 workshop participants; where consensus was achieved for 24 criteria and comments received on a proposed IVOS decision aid. Research recommendations include the use of evidence-based standardised IVOS criteria.

Discussion and Conclusion

This study achieved nationwide expert consensus on antimicrobial IVOS criteria for timely switch in the hospitalised adult population. For criteria operationalisation, an IVOS decision aid was developed. Further research is required to provide clinical validation of the consensus IVOS criteria and to expand this work into the paediatric and international settings.
Brain-wide measurement of synaptic protein turnover reveals localized plasticity during learning
Authors: Mohar Boaz , Michel Gabriela , Wang Yi-Zhi , Hernandez Veronica , Grimm Jonathan B. , Park Jin-Yong , Patel Ronak , Clarke Morgan , Brown Timothy A. , Bergmann Cornelius , Gebis Kamil K. , Wilen Anika P. , Liu Bian , Johnson Ricard , Graves Austin , Tchumatchenko Tatjana , Savas Jeffrey N. , Fornasiero Eugenio F. , Huganir Richard L. , Tillberg Paul , Lavis Luke D. , Svoboda Karel , Spruston Nelson
Published: November 2022

Synaptic plasticity underlies learning and memory by altering neuronal connections in response to experiences. However, the loci of learning-induced synaptic plasticity, and the degree to which plasticity is localized or distributed, remain largely unknown. We developed a method (DELTA) for mapping brain-wide changes in synaptic protein turnover with single-synapse resolution, based on Janelia Fluor dyes and HaloTag knock-in mice. During associative learning, the turnover of the ionotropic glutamate receptor GluA2, an indicator of synaptic plasticity, was enhanced in several brain regions, most markedly in the hippocampal area CA1. More broadly distributed increases in turnover of synaptic proteins were observed in response to environmental enrichment. In CA1, GluA2 stability was regulated in an input specific manner, with more turnover in layers containing input from CA3 compared to entorhinal cortex. DELTA will facilitate exploration of the molecular and circuit basis of learning and memory and other forms of adaptive and maladaptive plasticity at scales ranging from single synapses to the entire brain.
Dynamic spatial coding in parietal cortex mediates tactile-motor transformation
Authors: Klautke Janina , Foster Celia , Medendorp W. Pieter , Heed Tobias
Published: November 2022

Movements towards touch on the body require integrating tactile location and body posture information. Tactile processing and movement planning both rely on posterior parietal cortex (PPC) but their interplay is not understood. Here, human participants received tactile stimuli on their crossed and uncrossed feet, dissociating stimulus location relative to anatomy versus external space. Participants pointed to the touch or the equivalent location on the other foot, which dissociates sensory and motor locations. Multi-voxel pattern analysis of concurrently recorded fMRI signals revealed that tactile location was coded anatomically in anterior PPC but spatially in posterior PPC during sensory processing. After movement instructions were specified, PPC exclusively represented the movement goal in space, in regions associated with visuo-motor planning and with regional overlap for sensory, rule-related, and movement coding. Thus, PPC flexibly updates its spatial codes to accommodate rule-based transformation of sensory input to generate movement to environment and own body alike.
Degenerative changes are associated with severity of anterior cruciate ligament injury within the skeletally immature joint
Authors: Howe Danielle , Thompson Jacob D. , Teeter Stephanie D. , Easson Margaret , Barlow Olivia , Griffith Emily H. , Schnabel Lauren V. , Spang Jeffrey T. , Fisher Matthew B.
Published: November 2022

Anterior cruciate ligament (ACL) injuries are a major problem in the pediatric and adolescent populations. Some of these injuries extend only partially through the tissue cross-section; yet, there is limited data to inform clinical treatment of such partial tears. In particular, it is unknown how injury severity impacts long-term degenerative changes in the joint. Here, we leverage a skeletally immature preclinical porcine model to evaluate joint biomechanics and degeneration after partial (isolated anteromedial (AM) or posterolateral (PL) bundle) or complete ACL injury. Six months after injury, joint laxity increases were minimal after PL bundle injury, minor after AM bundle injury, and major after ACL injury. Joint degeneration (evaluated in the cartilage and meniscus) was minimal after PL bundle injury, moderate after AM bundle injury, and substantial after ACL injury. With subjects grouped by clinical Lachman grade (indicating the extent of joint destabilization), degeneration was associated with increasing grade, irrespective of injury type. These findings point to the importance of considering joint laxity as a factor when treating young patients, particularly those with partial ACL injuries.
Highly branched and complementary distributions of layer 5 and layer 6 auditory corticofugal axons in mouse
Authors: Issa Lina K. , Chandra Sekaran Nathiya Vaithiyalingam , Llano Daniel A.
Published: November 2022

The auditory cortex (AC) exerts a powerful, yet heterogeneous, effect on its subcortical targets. Auditory corticofugal projections emanate from distinct bands in layers 5 (L5) and 6 (L6), which have complementary anatomical and physiological properties. While several studies have suggested that corticofugal projections from L5 branch widely, others have suggested that there are multiple, mostly independent sets of L5 corticofugal projections. Even less is known about L6; no studies have examined whether the various L6 corticofugal projections are independent. Therefore, we examined branching patterns of L5 and L6 auditory corticofugal neurons, using the corticocollicular system as an index projection, using both traditional and novel approaches. We first confirmed that dual retrograde injections into the mouse inferior colliculus and auditory thalamus co-labeled subpopulations of L5 and L6 AC neurons. We then used an intersectional approach to selectively re-label L5 or L6 corticocollicular somata and found that both layers sent extensive branches to striatum, amygdala, superior colliculus, thalamus and nuclei of the lateral lemniscus. L5 corticocollicular axons also sent an unpaired projection to the superior olivary complex. Using a novel approach to separately label L5 and L6 axons in the same mouse, we found that L5/6 terminal distributions partially spatially overlapped and that a subset of giant terminals was only found in L5-derived axons. Overall, the high degree of branching and complementarity in the distributions of L5 vs. L6 axons suggest that corticofugal projections should be considered as two widespread systems of projections, rather than a collection of individual projections.
Can the brain strategically go on automatic pilot? An fMRI study investigating the effect of if-then planning on behavioral flexibility
Authors: van Timmeren Tim , Dzinalija Nadza , O’Doherty John P. , de Wit Sanne
Published: November 2022

People often have good intentions but fail to adhere to them. Implementation intentions, a form of strategic planning, can help people to close this intention-behavior gap. Their effectiveness has been proposed to depend on the mental formation of a stimulus-response association between a trigger and target behavior, thereby creating an ‘instant habit’. If implementation intentions do indeed lead to reliance on habitual control, then this may come at the cost of reduced behavioral flexibility. Furthermore, we would expect a shift from recruitment of corticostriatal brain regions implicated in goal-directed control towards habit regions. To test these ideas, we performed a functional MRI study in which participants received instrumental training supported by either implementation or goal intentions, followed by an outcome-revaluation to test reliance on habitual versus goal-directed control. We found that implementation intentions led to increased efficiency during training, as reflected in higher accuracy, faster reaction times, and decreased engagement of the anterior caudate. However, implementation intentions did not reduce behavioral flexibility when goals changed during the test phase, nor did it affect the underlying corticostriatal pathways. Additionally, this study showed that ‘slips of action’ towards devalued outcomes are associated with reduced activity in brain regions implicated in goal-directed control. In conclusion, our behavioral and neuroimaging findings suggest that strategic if-then planning does not lead to a shift from goal-directed towards habitual control.
A new method for quantifying APT and NOE(-3.5) using chemical exchange saturation transfer with double saturation powers (DSP-CEST)
Authors: Zhao Yu , Sun Casey , Zu Zhongliang
Published: November 2022

Purpose

Quantifications of amide proton transfer (APT) and nuclear Overhauser enhancement (NOE(−3.5)) mediated transfer with high specificity are challenging since their signals measured in a Z-spectrum are overlapped with confounding signals from direct water saturation (DS), semi-solid magnetization transfer (MT) and chemical exchange saturation transfer (CEST) of fast-exchange pools. In this study, based on two canonical CEST acquisitions with double saturation powers (DSP), a new data-postprocessing method is proposed to specifically quantify the effects of APT and NOE.

Methods

For CEST imaging with relatively low saturation powers, both the fast-exchange CEST effect and the semi-solid MT effect increase linearly withwhereas the slow-exchange APT/NOE(−3.5) effect has no such a dependence on, which is exploited to isolate the APT and NOE effects from the confounding signals in this study. After a mathematical derivation for the establishment of the proposed method, numerical simulations based on Bloch equations are then performed to demonstrate its specificity to detections of the APT and NOE effects. Finally, anin vivovalidation of the proposed method is conducted using an animal tumor model at a 4.7-T MRI scanner.

Results

The simulations show that DSP-CEST can quantify the effects of APT and NOE and substantially eliminate the confounding signals. The in vivo experiments demonstrate that the prosed DSP-CEST method is feasible for the imaging of tumors.

Conclusion

The data-postprocessing method proposed in this study can quantify the APT and NOE effects with considerably increased specificities and a reduced cost of imaging time.
Electron Videography of a Lipid–Protein Tango
Authors: Smith John W. , Carnevale Lauren N. , Das Aditi , Chen Qian
Published: November 2022

Biological phenomena ranging from enzymatic catalysis to synaptic transmission find their origins in the morphological fluctuations of biomolecules and biomolecular assemblies in liquid water. However, directly imaging these nanoscopic dynamics without probes or labels has been a fundamental methodological challenge. Here we developed an approach for ‘molecular videography’—combining liquid-phase transmission electron microscopy with molecular modelling—with which wefilmedthe nanoscale morphological fluctuations of individual, freely suspended, and unlabelled membrane protein nanodiscs in liquid. These unprecedented observations enabled us to quantify fingerprints of nanoscale lipid–protein interactions directly, revealing that nanodisc shape undulations are defined by the molecular mechanics of encircling scaffold proteins, whereas lipid interactions with a membrane protein reach across surprisingly long ranges to delineate dynamically modified membrane domains and increase nanodisc stability. Overall, this work illustrates an experimental approach to image, quantify, and understand biomolecular dynamics at the nanometre scale.
Determinants of regulatory compliance in health and social care services: a systematic review using the Consolidated Framework for Implementation Research
Authors: Dunbar Paul , Keyes Laura , Browne John P
Published: November 2022

Background

The delivery of high quality care is a fundamental goal for health systems worldwide. One policy tool to ensure quality is the regulation of minimum standards by an independent public authority. This systematic review seeks to identify determinants of compliance with such regulation in health and social care services.

Methods

Systematic searches were carried out on five electronic databases and grey literature sources. Quantitative, qualitative and mixed methods studies were eligible for inclusion. Titles and abstracts were screened by two reviewers independently. Determinants were identified from the included studies, extracted and allocated to constructs in the Consolidated Framework for Implementation Research (CFIR). The quality of included studies was appraised by two independent reviewers. The results were synthesised in a narrative review using the constructs of the CFIR as grouping themes.

Results

The search yielded 7,500 articles for screening, of which 157 were included. Most studies were quantitative designs in nursing home settings and were conducted in the United States. We found the following structural characteristics and compliance to be positively associated: smaller facilities (as measured by bed capacity); higher nurse-staffing levels; and lower staff turnover. A facility’s geographic location and compliance was also associated. We make no findings in respect of process determinants as qualitative studies were sparse, limiting investigation of the processes underlying regulatory compliance.

Conclusion

The literature in this field has focused to date on the structural attributes of compliant providers, perhaps because these are easier to measure, and has neglected more complex processes around the implementation of regulatory standards. A number of gaps, particularly in terms of qualitative work focused on the mechanisms involved in implementing regulations, are evident in the literature and further research in this area is needed to provide a clearer picture.
The dark side of channel loyalty programmes – managing tier demotion and deceitful behaviours
Authors: Cox David
Published: November 2022

Purpose

The purpose of this paper is to highlight the increased impact that academic research can gain when collaborating and sharing knowledge with corporate stakeholders and marketing practitioners, through the lens of managing tier demotion and deceitful behaviors in a channel marketing program. In particular, this paper highlights that true impact (defined as being meaningful change) can only be achieved when the research findings are operationalized and deployed as a business solution.

Design/methodology/approach

The approach taken in this paper is to challenge the traditional paradigm where field research is led by academics and places the corporate stakeholder at the centre of the research, where they have a leading role. This involves identifying, selecting, collaborating and engaging with key corporate stakeholders early in the research project, thus gaining their input and support so that the research recommendations secure the necessary funding to be successfully deployed.

Findings

The authors highlight two research projects focused on tier demotion and disengagement and deceitful behaviour driven from incentivise and gamifying learning. A new tier demotion process involving a highly customised customer care charter has been deployed and resulted in a re-engagement rate of 81% from those channel partners who were demoted (a rise from 40%). Deceitful behaviour detection tools were also deployed, resulting in 4,300 possible deceptive cases being investigated and resolved in Year 1, reducing to 451 in Year 6.

Research limitations/implications

The change in research approach where field research places the corporate stakeholder at the centre of the research will add to the complexity of the research project, impact on timelines and introduce more stakeholders into the research team, which may have wider implications to the original research goal.

Practical implications

This transition will require academics to know more about their corporate partner, the corporate landscape, adding to the complexity of the research project and cede some control over the project.

Originality/value

Whilst academic research contributes to a body of theoretical concepts, equally important is bringing to life the research findings to show actual impact and meaningful change within the research setting. The present research approach has since been applied to multiple channel loyalty programmes across numerous industries increasing revenue and driving the success of the respective programmes. One key learning would be to engage more corporate stakeholders as part of the research project from the outset. The author neglected the legal and trust and compliance team and they had a significant impact on what findings were eventually implemented.