bluuebunny/crossref_metadata_2025 huggingface dataset:
Reference intervals of amino acid for newborns in North Asia: Diagnosing Inborn Errors of Metabolism Using Liquid Chromatography Tandem Mass Spectrometry
Authors: Bairova T.A. , Belskikh A.V. , Atalyan A.V. , Bugun O.V. , Nemchinova N.V. , Rychkova L.V.
Published: April 2021

Aim

To establish reference intervals (RI) for blood amino acids (AA) in healthy newborns of North Asia measured by liquid chromatography tandem mass spectrometry (LC-MS/MS) and evaluate their differences from respective reference values for newborns from other populations.

Objectives

A cross-sectional study of 381 healthy newborns was conducted. De-identified dried blood spots annotated by age, birth-weight, and sex were obtained from 381 healthy newborns aged 0–7 days. Data was collected from April to May of 2020.

Methods

Dried blood spots collected from filtered paper were used to analyze and measure of 13 derivatized amino acids using LC-MS/MS method. Nonparametric statistical approaches were used to generate 2.5th–97.5th percentile distributions for newborns in North Asia in accordance with CLSI EP28-A3c.

Results

Reference intervals (RI) for phenylalanine, tyrosine, citrulline, alanine, ornithine, proline in North Asian newborns differ slightly from those of newborns in other countries around the world. This allows for the use of universal RI in the diagnosis of congenital metabolic disorders of the indicated amino acids (AA) in different populations around the world. The RI for branched-chain essential AA (valine, leucine, and isoleucine are metabolic criteria for maple syrup disease) are higher in North Asian infants as compared to infants in other populations. In addition, the RI for arginine, aspartic acid, and glutamic acid in North Asian newborns are higher than in newborns in other countries around the world. In our study, the RI for methionine in newborns were lower than in many countries worldwide [McHugh D, 2011]. For optimal clinical practice, RI for certain AA in newborns (valine, leucine, isoleucine, methionine, arginine, aspartic acid, glutamic acid) should be determined for specific populations-further augmenting the diagnosis of inborn errors of metabolism.
Combining Indoor Positioning Using Wi-Fi Round-trip Time with Dust Measurement in the Field of Occupational Health
Authors: Ando Hajime , Sekoguchi Shingo , Ikegami Kazunori , Yoshitake Hidetaka , Baba Hiroka , Myojo Toshihiko , Ogami Akira
Published: April 2021

[Objectives]

The measurement of personal exposure has been attracting increased attention for the control of hazardous substances. Although the combination of location information and personal exposure is considered highly useful, to-this-date, no simple or highly accurate indoor positioning method has been presented. Recently, Wi-Fi round-trip time (RTT) has attracted scientific attention as a highly accurate indoor positioning method. In this study, we investigated the possibility of the use of a combination of this indoor positioning method and small-dust sensors.

[Methods]

We measured indoor positioning and dust concentration three times during a walk in a simulated factory. A wearable particle monitor was worn on both upper arms, and a smartphone was attached to the helmet for positioning. An ultrasonic humidifier was used to spray sodium chloride solution in the factory. The measurements were conducted three times on different routes on foot (Experiment A, B, C).

[Results]

In indoor positioning, the error percentages, i.e., measurements that were outside the correct measurement area were 7% (49 s/700 s) in Experiment A, 2.3% (15 s/660 s) in Experiment B, and 7.8% (50 s/645 s) in Experiment C. The dust measurements were also taken without interruption. A heat map was created based on the results of both measurements.

[Conclusions]

Wi-Fi RTT was able to implement indoor positioning with high accuracy. Thus, the combination of indoor positioning and various sensors can be considered useful for occupational health.
Assess the Benefit of Interventional Radiologist Performing an Image Guided Biopsy
Authors: Rozenberg Radu , Shahi Niharika , Burute Nishigandha , Kotb Ahmed , Shahrour Walid , Kisselgoff David , van der Pol Christian B. , Shuster Anatoly
Published: April 2021

Purpose

To determine whether there is any benefit in diagnostic accuracy, reduction in periprocedural complications, when image-guided biopsies are performed by interventional radiologist as compared to a general radiologist or other physician without prior special interventional training.

Patients and Methods

This study is a retrospective chart review of all consecutive patients that underwent imaging-guided core biopsies during one year in our hospital. Information collected included: patient age, gender, coagulation status, organ biopsied, imaging modality and equipment used, number of samples obtained, post-biopsy complications, histopathological results, and previous interventional training of a physician performing the biopsy. The quantitative data from patient charts was analyzed using a statistical program, Statistical Package for Social Sciences (SPSS).

Results

449 patients were included in this study: 132 were performed using Computed Tomography guidance and 317 were performed under ultrasound guidance. The success rate of core biopsies was compared between different specialists, and was measured based on periprocedural complications, necessity of medical intervention and/or hospitalization after biopsy procedure, true positive histopathology results, and need to perform repeat biopsy of the same lesion owing to inconclusive results. Overall, IR had a success rate of 88.13%, non-IR had a success rate of 61.07%, and nephrologists had a success rate of 77.65%. The post biopsy complication rates were 5.48%, 27.52%, and 17.65% for procedures performed by IR, non-IR, and nephrologists respectively.

Conclusion

Our study shows an overall higher success rate and improved patient outcome when image guided biopsies were performed by IR, as compared to other non-IR physicians.
A Real World Evaluation of the safety and immunogenicity of the Covishield vaccine, ChAdOx1 nCoV- 19 Corona Virus Vaccine (Recombinant) in Health Care Workers (HCW) in National Capital Region (NCR) of India: A preliminary report
Authors: Kataria Sushila , Sharma Pooja , Deswal Vikas , Kumar Kuldeep , Singh Manish , Alam Sazid , Gupta Vaibhav , Singh Padam , Phogat Rashmi , Sarma Smita , Patil Nipun , Dutt Rohit , Saxena Renu , Trehan Naresh
Published: April 2021

Background

The SARS-CoV-2 pandemic has severely impacted health systems, economic and social progress globally in 2020. The rollout of vaccines in several parts of the world is being hailed as a solution to the crisis. With newer and more virulent serotypes on the horizon and limited vaccine available, evaluation of safety and immunogenicity is critical for rationalization of vaccine use in public health.

Objective

To evaluate real world safety, and, immunogenicity of the Covishield vaccine, ChAdOx1 nCoV-19 Corona Virus Vaccine (Recombinant) in Health Care Workers (HCW) during the national vaccine roll out in the NCR, New Delhi. The safety is evaluated through Adverse Events and Serious Adverse Events reported though enhanced pharmacovigilance protocols, and, the immunogenicity by quantitative determination of anti-S1 and anti-S2 specific IgG antibodies to SARS-CoV-2 in serum samples collected before the receipt of the vaccine and 14 days after dose 1, using the fully automated LIAISON® SARS-CoV-2 S1/S2 IgG test using the chemiluminescence immunoassay (CLIA)

Results

In the two weeks after immunization with the Covishield vaccine {ChAdOx1 nCoV-19 Corona Virus Vaccine (Recombinant)}, none of the 1638 evaluated participants reported any serious adverse events (ie require hospitalization or emergency room visit). Solicited adverse events reported via daily diary cards included pain (62.7%) and soreness (24.1%) at injection site as most common, whereas fever (48.4%), headache (43.4%), myalgia (38.4%), fatigue (33.4%), joint pain (27.0%) and nausea (16.0%) were most common solicited systemic adverse events on day 1. Majority of local and systemic adverse events were seen in first 2 days post vaccination and thereafter they resolved. Lesser reactogenicity was observed in subjects with age >50 years. No major difference was observed in adverse events when subjects were stratified based on history of COVID 19 disease or baseline seropositivity. In our study serostatus improved from 48.2% positive at baseline to 79.0% positive 2 weeks following first dose of vaccination. After first dose of vaccination overall higher percentage (98.2%) of seropositivity rates were observed in those with past history of COVID 19 disease

Conclusion

The Covishield vaccine {ChAdOx1 nCoV-19 Corona Virus Vaccine (Recombinant)}, was safe and reported mild self limiting adverse events over 2-4 days and had an good early (within 2 weeks) seroresponse. This holds the promise of far reaching impact on vaccine availability for a larger population and thereby providing a widespread coverage.
Diagnostic accuracy of SARS-CoV-2 rapid antigen detection testing in symptomatic and asymptomatic children in the clinical setting
Authors: L’Huillier Arnaud G. , Lacour Matthieu , Sadiku Debora , Gadiri Mehdi A. , De Siebenthal Loraine , Schibler Manuel , Eckerle Isabella , Pinösch Selina , Kaiser Laurent , Gervaix Alain , Glangetas Alban , Galetto-Lacour Annick , Lacroix Laurence
Published: April 2021

Importance

Antigen-based rapid diagnostic tests (RDTs) have shown good sensitivity for SARS-CoV-2 detection in adults and are used in children despite the lack data from children.

Objective

We evaluated the diagnostic performance of the Panbio™-COVID-19 Ag Rapid Test Device (P-RDT) in symptomatic and asymptomatic children against reverse-transcription polymerase chain reaction (RT-PCR) on nasopharyngeal swabs (NPS).

Design

Prospective diagnostic study from 11.2020 to 03.2021.

Setting

Single-center.

Participants

Consecutive symptomatic and asymptomatic participants 0-16yo.

Intervention

Two NPS for both RT-PCR and P-RDT.

Main outcome

P-RDT sensitivity and specificity.

Results

Eight-hundred and twenty-two participants completed the study, of which 533 (64.9%) were symptomatic. Among the 119 (14.5%) RT-PCR positive patients, the overall P-RDT sensitivity was 0.66 (95%CI 0.57-0.74). Mean viral load (VL) was higher among P-RDT positive than negative ones (p<0.001). Sensitivity was 0.87 in specimens with VL>1.0E6 copies/mL (95%CI 0.87-1.00), which is the accepted cut-off for the presence of infectious virus, and decreased to 0.67 (95%CI 0.59-0.76) for specimens >1.0E3 copies/mL.

Among symptomatic participants, the P-RDT displayed a sensitivity of 0.73 (95%CI 0.64-0.82), which peaked at 1.00 at 2 days post onset of symptoms (DPOS; 95%CI 1.00-1.00), then decreased to 0.56 (95%CI 0.23-0.88) at 5 DPOS. There was a trend towards lower P-RDT sensitivity in symptomatic children <12 years (0.62 [95%CI 0.45-0.78]) versus ≥12 years (0.80 [95%CI 0.69-0.91]; p=0.09). VL which was significantly lower in asymptomatic participants than in symptomatic ones (p<0.001). The P-RDT displayed a sensitivity of 0.43 (95%CI 0.26-0.61).

Specificity was 1.00 in symptomatic and asymptomatic children (95%CI 0.99-1.00).

Conclusion and relevance

The overall respective 73% and 43% sensitivities of P-RDT in symptomatic and asymptomatic children was below the 80% cut-off recommended by the World Health Organization. These findings are likely explained by lower VLs in children at the time of diagnosis. As expected, we observed a direct correlation between VL and P-RDT sensitivity as well as variation of sensitivity according to DPOS, a major determinant of VL. These data highlight the limitations of RDTs both in symptomatic and asymptomatic children, with the potential exception in early symptomatic children ≥12yrs where sensitivity reached 80%.
A novel proneural function of Asense, integrated with of L’sc and Notch, promotes the Neuroepithelial to Neuroblast transition
Authors: Martin Mercedes , Shaikh Mirja N. , Gutierrez-Avino Francisco , Tejedor Francisco J.
Published: April 2021

In the developingDrosophilaoptic lobe, neuroepithelial (NE) cells are transformed progressively into neurogenic progenitors called neuroblasts (NBs). The progenitors undergoing this transition are identified by the expression of the Acheate Scute Complex (AS-C) factor Lethal of Scute (L’sc).

Here we found that Asense (Ase), another AS-C factor, presents a peak of expression in the cells neighboring those transition L’sc expressing cells. This peak of Ase identifies a new transition step and it is necessary and sufficient to promote the NE to NB transition. Thus, our data provide the first direct evidence for a proneural role of Ase in CNS neurogenesis. Furthermore, we found that the peak of Ase is induced in a non-cell autonomous manner by L’sc through the activation of Notch signaling in the adjacent cells. This suggests that the two classic proneural activities, promoting neurogenesis and Notch signaling, have been split between Ase and L’sc. Thus, our data fit with a model in which the key proneural role of Ase is integrated with Notch and L’sc activities, facilitating the progressive transformation of NE cells into NBs.

SUMMARY STATEMENT

The switch of progenitor cells towards neuron production is crucial for proper brain development. The transcription factor Asense promotes this transition through a mechanism integrated with timing and neurogenic signals.
ODACH: A One-shot Distributed Algorithm for Cox model with Heterogeneous Multi-center Data
Authors: Luo Chongliang , Duan Rui , Chen Yong
Published: April 2021

Objective

We developed and evaluated a privacy-preserving One-shot Distributed Algorithm for Cox model to analyze multi-center time-to-event data without sharing patient-level information across sites, while accounting for heterogeneity across sites by allowing site-specific baseline hazard functions and feature distributions.

Materials and Methods

We constructed a surrogate likelihood function to approximate the Cox log partial likelihood function which is stratified by site, using patient-level data from a single site and aggregated information from other sites. The ODAC estimator was obtained by maximizing the surrogate likelihood function. We evaluated and compare the performance of ODACH with meta-analysis by extensive numerical studies.

Results

The simulation study showed that ODACH provided estimates close to the pooled estimator, which is obtained by directly analyzing patient-level data from all sites via a stratified Cox model. The relative bias was <1% across all scenarios. As a comparison, the meta-analysis estimator, which was obtained by the inverse variance weighted average of the site-specific estimates, had substantial bias when the event rate is <5%, with the relative bias reaching 12% when the event rate is 1%.

Conclusions

ODACH is a privacy-preserving and communication-efficient method for analyzing multi-center time-to-event data, which allows the baseline hazard functions as well as the distribution of covariate variables to vary across sites. It provides estimates that is close to the pooled estimator and substantially outperforms the meta-analysis estimator when the event is rare. It is thus extremely suitable for studying rare events with heterogeneous baseline hazards across sites in a distributed manner.
A Simplified Point-of-Care Lung Ultrasound Protocol to Detect Coronavirus Disease 2019 in Inpatients: A Prospective Observational Study
Authors: Heyne Thomas F. , Geisler Benjamin P. , Negishi Kay , Choi Daniel , Saud Ahad A. Al , Marinacci Lucas X. , Smithedajkul Patrick Y. , Devaraj Lily R. , Little Brent P. , Mendoza Dexter P. , Flores Efren J. , Petranovic Milena , Toal Steven P. , Shokoohi Hamid , Liteplo Andrew S.
Published: April 2021

Objectives

To assess the diagnostic performance of lung point-of-care ultrasound (POCUS) compared to either a positive nucleic acid test (NAT) or a COVID-19-typical pattern on computed tomography (CT) and to evaluate opportunities to simplify a POCUS algorithm.

Methods

Hospital-admitted adult inpatients with (1) either confirmed or suspected COVID-19 and (2) a completed or ordered CT within the preceding 24 hours were recruited. Twelve lung zones were scanned with a handheld POCUS machine. POCUS, CT, and X-ray (CXR) images were reviewed independently by blinded experts. A simplified POCUS algorithm was developed via machine learning.

Results

Of 79 enrolled subjects, 26.6% had a positive NAT and 31.6% had a CT typical for COVID-19. The receiver operator curve (ROC) for a 12-zone POCUS protocol had an area under the curve (AUC) of 0.787 for positive NAT and 0.820 for typical CT. A simplified four-zone protocol had an AUC of 0.862 for typical CT and 0.862 for positive NAT. CT had an AUC of 0.815 for positive NAT; CXR had AUCs of 0.793 for positive NAT and 0.733 for typical CT. Performance of the four-zone protocol was superior to CXR for positive NAT (p=0.0471). Using a two-point cutoff system, the four-zone POCUS protocol had a sensitivity of 0.920 and 0.904 compared to CT and NAT, respectively, at the lower cutoff; it had a specificity of 0.926 and 0.948 at the higher cutoff, respectively.

Conclusion

POCUS outperformed CXR to predict positive NAT. POCUS could potentially replace other chest imaging for persons under investigation for COVID-19.
On whether therapeutic plasma exchange is an effective cure against severe/critical COVID-19 pneumonia
Authors: Cegolon Luca , Einollahi Behzad , Imanizadeh Sina , Rezapour Mohammad , Javanbakht Mohammad , Nikpouraghdam Mohammad , Abolghasemi Hassan , Mastrangelo Giuseppe
Published: April 2021

Background

There is a risk of novel mutations of SARS-CoV-2 that may render COVID-19 resistant to most of the therapies, including antiviral drugs. The evidence around the application of therapeutic plasma exchange (TPE) for the management of critically ill COVID-19 patients is still provisional and further investigations are needed to confirm its eventual beneficial effects.

Methods

We therefore carried out a single-centered retrospective observational non-placebo-controlled trial enrolling 73 inpatients from Baqiyatallah Hospital in Tehran (Iran) with diagnosis of COVID-19 pneumonia confirmed by real-time polymerase chain reaction (RT-PCR) on nasopharyngeal swabs and high-resolution computerized tomography chest scan. These patients were broken down into two groups: Group 1 (30 patients) receiving standard of care (corticosteroids, ceftriaxone, azithromycin, pantoprazole, hydroxychloroquine, lopinavir/ritonavir); and Group 2 (43 patients) receiving the above regimen plus TPE (replacing 2 liter of patients’ plasma by a solution, 50% of normal plasma and 50% of albumin at 5%) administered according to various time schedules. The follow-up time was 30 days and all-cause mortality was the endpoint.

Results

Deaths were 6 (14%) in Group 2 and 14 (47%) in Group 1. However, different harmful risk factors prevailed among patients not receiving TPE rather than being equally split between the intervention and control group. We used an algorithm of Structural Equation Modeling (of STATA) to summarize a large pool of potential confounders into a single score (called with the descriptive name “severity”). Disease severity was significantly (Wilkinson rank sum test p-value=0.0000) lower among COVID-19 patients undergoing TPE (median: −2.82; range: −5.18; 7.96) as compared to those non receiving TPE (median: −1.35; range: −3.89; 8.84), confirming that treatment assignment involved a selection bias of patients according to the severity of COVID-19 at hospital admission. The adjustment for confounding was carried out using severity as covariate in Cox regression models. The univariate Hazard Ratio (HR) of 0.68 (95%CI: 0.26; 1.80; p=0.441) for TPE turned to 1.19 (95%CI: 0.43; 3.29; p=0.741) after adjusting for severity.

Conclusions

The lower mortality observed among patients receiving TPE was due to a lower severity of COVID-19 rather than TPE effects.

TRIAL REGISTRATION

IRCT registration number: IRCT20080901001165N58 (Iranian Registry of Clinical Trials)

Registration date: 2020-05-27, 1399/03/07 (retrospectively registered)
Analyzing the effect of relaxing restriction on the COVID-19 outbreak for some US states
Authors: Demir Mahir , Aslan ibrahim Halil , Lenhart Suzanne
Published: April 2021

The ongoing pandemic disease COVID-19 has caused worldwide social and financial disruption. As many countries are engaged in designing vaccines, the harmful second and third waves of COVID-19 have already appeared in many countries. To investigate changes in transmission rates and the effect of social distancing in the USA, we formulate a system of ordinary differential equations using data of confirmed cases and deaths in these states: California, Texas, Florida, Georgia, Illinois, Louisiana, Michigan, and Missouri in the USA to be able to investigate changes in transmission rates of the outbreak and effect of social distancing. Our models and the corresponding parameter estimations show social distancing reduces the transmission by 60% to 90%, and thus obeying the movement restriction rules plays a crucial rule to reduce the magnitudes of the outbreak waves. Our analysis shows the current management restrictions do not sufficiently slow the disease propagation.
Novel genome characteristics contribute to the invasiveness ofPhragmites australis(common reed)
Authors: Oh Dong-Ha , Kowalski Kurt P. , Quach Quynh N. , Wijesinghege Chathura , Tanford Philippa , Dassanayake Maheshi , Clay Keith
Published: April 2021

The rapid invasion of the non-nativePhragmites australis(Poaceae, subfamily Arundinoideae) is a major threat to native ecosystems in North America. We describe a 1.14 Gbp reference genome forP. australisand compare invasive (ssp.australis) and native (ssp.americanus) genotypes collected across the Laurentian Great Lakes to deduce genomic bases driving its invasive success. We report novel genomic features including a lineage-specific whole genome duplication, followed by gene loss and preferential retention of genes associated with transcription factors and regulatory functions in the remaining duplicates. The comparative transcriptomic analyses revealed that genes associated with biotic stress and defense responses were expressed at a higher basal level in invasive genotypes, but the native genotypes showed a stronger induction of defense responses following fungal inoculation. The reference genome and transcriptomes, combined with previous ecological and environmental data, support the development of novel, genomics-assisted management approaches for invasivePhragmites.
Nanoconfinement of Microvilli Alters Gene Expression and Boosts T cell Activation
Authors: Aramesh Morteza , Stoycheva Diana , Sandu Ioana , Ihle Stephan J. , Zünd Tamara , Shiu Jau-Ye , Forró Csaba , Asghari Mohammad , Bernero Margherita , Lickert Sebastian , Oxenius Annette , Vogel Viola , Klotzsch Enrico
Published: April 2021

T cells sense and respond to their local environment at the nanoscale by forming small actin-rich protrusions, called microvilli, which play critical roles in signaling and antigen recognition, particularly at the interface with the antigen presenting cells. However, the mechanisms by which microvilli contribute to cell signaling and activation is largely unknown. Here, we present a tunable engineered system that promotes microvilli formation and T cell signaling via physical stimuli. We discovered that nanoporous surfaces favored microvilli formation, and markedly altered gene expression in T cells and promoted their activation. Mechanistically, confinement of microvilli inside of nanopores leads to size-dependent sorting of membrane-anchored proteins, specifically segregating CD45 phosphatases and T cell receptors (TCR) from the tip of the protrusions when microvilli are confined in 200 nm pores, but not in 400 nm pores. Consequently, formation of TCR nanoclustered hotspots within 200 nm pores, allows sustained and augmented signaling that prompts T cell activation even in the absence of TCR agonists. The synergistic combination of mechanical and biochemical signals on porous surfaces presents a straightforward strategy to investigate the role of microvilli in T cell signaling as well as to boost T cell activation and expansion for application in the growing field of adoptive immunotherapy.
Contrasting impacts of environmental variability on the breeding biology of two sympatric small procellariiform seabirds in south-eastern Australia
Authors: Eizenberg Yonina H. , Fromant Aymeric , Lec’hvien Arnaud , Arnould John P.Y.
Published: April 2021

Seabirds play a vital role in marine ecosystems and are determinant sentinels of the productivity of their environments. The long-term study of their breeding biology and their responses to environmental variations can be used to monitor the effects of climate change on marine fauna. However, the ecological and physiological differences among seabirds induce a large range of responses complicating our understanding of the effects of environmental changes on marine ecosystems. The present study investigated the impact of environmental variability on breeding biology in two sympatric small Procellariiform species, the fairy prion (Pachyptila turtur) and the common diving petrel (Pelecanoides urinatrix), over four reproductive seasons (2017-2020) in Bass Strait, south-eastern Australia. Marine heatwaves had a negative effect on chick growth, breeding success, and induced a delay in laying dates in both species. While fairy prions maintained a relatively high breeding success and broadly constant breeding phenology, common diving petrels delayed the start of the breeding season by up to 50 days and experienced dramatic collapses in breeding success in years of high marine heat wave occurrence. The high wing loading and absence of stomach oils in the common diving petrel are likely to have limited the capacity of this species to increase foraging effort in years of low food availability.
The Psychometrics of Automatic Speech Recognition
Authors: Weerts Lotte , Rosen Stuart , Clopath Claudia , Goodman Dan F. M.
Published: April 2021

Deep neural networks have had considerable success in neuroscience as models of the visual system, and recent work has suggested this may also extend to the auditory system. We tested the behaviour of a range of state of the art deep learning-based automatic speech recognition systems on a wide collection of manipulated sounds used in standard human psychometric experiments. While some systems showed qualitative agreement with humans in certain tests, in others all tested systems diverged markedly from humans. In particular, all systems used spectral invariance, temporal fine structure and speech periodicity differently from humans. We conclude that despite some promising results, none of the tested automatic speech recognition systems can yet act as a strong proxy for human speech recognition. However, we note that the more recent systems with better performance also tend to better match human results, suggesting that continued cross-fertilisation of ideas between human and automatic speech recognition may be fruitful. Our open source toolbox allows researchers to assess future automatic speech recognition systems or add additional psychoacoustic measures.
Hexavalent Sperm-Binding IgG Antibody Released from Self-Dissolving Vaginal Film Enables Potent, On-Demand Non-Hormonal Female Contraception
Authors: Shrestha Bhawana , Vincent Kathleen , Schaefer Alison , Zhu Yong , Vargas Gracie , Motamedi Massoud , Swope Kelsi , Morton Josh , Simpson Carrie , Pham Henry , Brennan Miles B. , Pauly Michael H. , Zeitlin Larry , Bratcher Barry , Whaley Kevin J. , Moench Thomas R. , Lai Samuel K.
Published: April 2021

Non-hormonal products for on-demand contraception are a global health technology gap, motivating us to pursue the use of sperm-binding monoclonal antibodies as a strategy to enable safe, effective, desirable, on-demand contraception. Here, using cGMP-compliant Nicotiana-expression system, we produce an ultra-potent sperm-binding IgG antibody possessing 6 Fab arms per molecule that bind a well-established contraceptive antigen target, CD52g. We term this hexavalent antibody “Fab-IgG-Fab” (FIF) to reflect its molecular orientation. The Nicotiana-produced FIF exhibits at least 10-fold greater sperm agglutination potency and kinetics than the parent IgG, while preserving Fc-mediated trapping of individual spermatozoa in mucus. We formulate the Nicotiana-produced FIF into a polyvinyl alcohol-based water-soluble contraceptive film, and evaluate its potency in reducing progressively motile sperm in the sheep vagina. Two minutes after vaginal instillation of human semen, no progressively motile sperm are recovered from the vaginas of sheep receiving FIF-Film. In contrast, high numbers of progressively motile sperm are recovered from sheep receiving a placebo film control. Our work supports the potential of highly multivalent contraceptive antibodies to provide safe, effective, on-demand non-hormonal contraception.
Repair of critical-size porcine craniofacial bone defects using a collagen-polycaprolactone composite biomaterial
Authors: Dewey Marley J. , Milner Derek J. , Weisgerber Daniel , Flanagan Colleen L. , Rubessa Marcello , Lotti Sammi , Polkoff Kathryn M. , Crotts Sarah , Hollister Scott J. , Wheeler Matthew B. , Harley Brendan A.C.
Published: April 2021

Regenerative medicine approaches for massive craniomaxillofacial bone defects face challenges associated with the scale of missing bone, the need for rapid graft-defect integration, and challenges related to inflammation and infection. Mineralized collagen scaffolds have been shown to promote mesenchymal stem cell osteogenesis due to their porous nature and material properties, but are mechanically weak, limiting surgical practicality. Previously, these scaffolds were combined with 3D-printed polycaprolactone mesh to form a scaffold-mesh composite to increase strength and promote bone formation in sub-critical sized porcine ramus defects. Here, we compare the performance of mineralized collagen-polycaprolactone composites to the polycaprolactone mesh in a critical-sized porcine ramus defect model. While there were no differences in overall healing response between groups, our data demonstrated broadly variable metrics of healing regarding new bone infiltration and fibrous tissue formation. Abscesses were present surrounding some implants and polycaprolactone polymer was still present after 9-10 months of implantation. Overall, while there was limited successful healing, with 2 of 22 implants showed substantial levels of bone regeneration, and others demonstrating some form of new bone formation, the results suggest targeted improvements to improve repair of large animal models to more accurately represent craniomaxillofacial bone healing. Notably, strategies to increase osteogenesis throughout the implant, modulate the immune system to support repair, and employ shape-fitting tactics to avoid implant micromotion and resultant fibrosis. Improvements to the mineralized collagen scaffolds involve changes in pore size and shape to increase cell migration and osteogenesis and inclusion or delivery of factors to aid vascular ingrowth and bone regeneration.
Drug repurposing to face Covid-19: Celastrol, a potential leading drug capable of inhibiting SARS-CoV-2 replication and induced inflammation
Authors: Fuzo Carlos A. , Martins Ronaldo B. , Fraga-Silva Thais F.C. , Amstalden Martin K. , De Leo Thais Canassa , Souza Juliano P. , Lima Thais M. , Faccioli Lucia H. , França Suzelei C. , Bonato Vania L.D. , Arruda Eurico , Dias-Baruffi Marcelo
Published: April 2021

The global emergence of Covid-19 has caused huge human casualties. Clinical manifestations of the disease vary from asymptomatic to lethal, and the symptomatic form can be associated with cytokine storm and non-homeostatic inflammatory response. In face of the urgent demand for effective drugs to treat Covid-19, we have searched for candidate compounds using a drug repurposing approach based onin silicoanalysis followed by biological validation. Here we identified celastrol, a pentacyclic triterpene isolated fromTripterygium wilfordiiHook F – a plant used in traditional Chinese medicine – as one of the best compounds out of 39 repurposed drug candidates. Celastrol reverted gene expression signature from SARS-CoV-2-infected cells; bound with high-affinity energy to viral molecular targets such as main protease (Mpro) and receptor-biding domain (RBD); inhibited SARS-CoV-2 replication in monkey (Vero and Vero-ACE2) and human (Caco-2 and Calu-3) cell lines; and decreased interleukin-6 (IL-6) secretion in SARS-CoV-2-infected human cell lines. Interestingly, celastrol acted in a concentration-dependent manner, with undetectable signs of cytotoxicity. Therefore, celastrol is a promising lead drug candidate to treat Covid-19 due to its ability to suppress SARS-CoV-2 replication and IL-6 production in infected cells, two critical events in the pathophysiology of this disease.
Structures of a deAMPylation complex rationalise the switch between antagonistic catalytic activities of FICD (14/96/109)
Authors: Perera Luke A. , Preissler Steffen , Zaccai Nathan R , Prévost Sylvain , Devos Juliette M , Haertlein Michael , Ron David
Published: April 2021

The endoplasmic reticulum (ER) Hsp70 chaperone BiP is regulated by AMPylation, a reversible inactivating post-translational modification. Both BiP AMPylation and deAMPylation are catalysed by a single ER-localised enzyme, FICD. Here we present long-sought crystallographic and solution structures of a deAMPylation Michaelis complex formed between mammalian AMPylated BiP and FICD. The latter, via its tetratricopeptide repeat domain, binds a surface that is specific to ATP-state Hsp70 chaperones, explaining the exquisite selectivity of FICD for BiP’s ATP-bound conformation both when AMPylating and deAMPylating Thr518. The eukaryotic deAMPylation mechanism thus revealed, rationalises the role of the conserved Fic domain Glu234 as a gatekeeper residue that both inhibits AMPylation and facilitates hydrolytic deAMPylation catalysed by dimeric FICD. These findings point to a monomerisation-induced increase in Glu234 flexibility as the basis of an oligomeric state-dependent switch between FICD’s antagonistic activities, despite a similar mode of engagement of its two substrates — unmodified and AMPylated BiP.
Expansion microscopy facilitates quantitative super-resolution studies of cytoskeletal structures in kinetoplastid parasites
Authors: Gorilak Peter , Pružincová Martina , Vachova Hana , Olšinová Marie , Varga Vladimir
Published: April 2021

Expansion microscopy (ExM) has become a powerful super-resolution method in cell biology. It is a simple, yet robust approach, which does not require any instrumentation or reagents beyond those present in a standard microscopy facility. In this study, we used kinetoplastid parasitesTrypanosoma bruceiandLeishmania major, which possess a complex, yet well-defined microtubule-based cytoskeleton, to demonstrate that this method recapitulates faithfully morphology of structures as previously revealed by a combination of sophisticated electron microscopy (EM) approaches. Importantly, we also show that due to rapidness of image acquisition and 3D reconstruction of cellular volumes ExM is capable of complementing EM approaches by providing more quantitative data. This is demonstrated on examples of less well-appreciated microtubule structures, such as the neck microtubule ofT. bruceior the pocket, cytosolic, and multivesicular tubule-associated microtubules ofL. major. We further demonstrate that ExM enables identifying cell types rare in a population, such as cells in mitosis and cytokinesis. 3D reconstruction of an entire volume of these cells provided details on morphology of the mitotic spindle and the cleavage furrow. Finally, we show that established antibody markers of major cytoskeletal structures function well in ExM, which together with the ability to visualize proteins tagged with small epitope tags will facilitate studies of the kinetoplastid cytoskeleton.
Training-Induced Circuit-Specific Excitatory Synaptogenesis is Required for Effort Control
Authors: Severino Francesco Paolo Ulloa , Lawal Oluwadamilola , Sakers Kristina , Wang Shiyi , Kim Namsoo , Friedman Alexander , Johnson Sarah , Sriworarat Chaichontat , Hughes Ryan , Soderling Scott , Kim Il Hwan , Yin Henry , Eroglu Cagla
Published: April 2021

Synaptogenesis is essential for circuit development; however, it is unknown whether it is critical for the establishment and performance of goal-directed voluntary behaviors. Here, we show that operant-conditioning via lever-press for food reward-training in mice induces excitatory synapse formation onto a subset of Anterior Cingulate Cortex neurons projecting to the dorsomedial striatum (ACC->DMS). Training-induced synaptogenesis is controlled by the Gabapentin/Thrombospondin receptor α2δ-1, which is an essential neuronal protein for proper intracortical excitatory synaptogenesis. Using germline and conditional knockout mice, we found that deletion of α2δ-1 in the adult ACC->DMScircuit diminishes training-induced excitatory synaptogenesis. Surprisingly, this manipulation did not impact learning but instead resulted in a profound increase in effort exertion without affecting sensitivity to reward value or changing contingencies. Bidirectional optogenetic manipulation of ACC->DMSneurons rescued or phenocopied the behaviors of the α2δ-1 cKO mice highlighting the importance of synaptogenesis within this cortico-striatal circuit in regulating effort exertion.
Immune Correlates of Protection by mRNA-1273 Immunization against SARS-CoV-2 Infection in Nonhuman Primates
Authors: Corbett Kizzmekia S. , Nason Martha C. , Flach Britta , Gagne Matthew , O’ Connell Sarah , Johnston Timothy S. , Shah Shruti N. , Edara Venkata Viswanadh , Floyd Katharine , Lai Lilin , McDanal Charlene , Francica Joseph R. , Flynn Barbara , Wu Kai , Choi Angela , Koch Matthew , Abiona Olubukola M. , Werner Anne P. , Alvarado Gabriela S. , Andrew Shayne F. , Donaldson Mitzi M. , Fintzi Jonathan , Flebbe Dillon R. , Lamb Evan , Noe Amy T. , Nurmukhambetova Saule T. , Provost Samantha J. , Cook Anthony , Dodson Alan , Faudree Andrew , Greenhouse Jack , Kar Swagata , Pessaint Laurent , Porto Maciel , Steingrebe Katelyn , Valentin Daniel , Zouantcha Serge , Bock Kevin W. , Minai Mahnaz , Nagata Bianca M. , Moliva Juan I. , van de Wetering Renee , Boyoglu-Barnum Seyhan , Leung Kwanyee , Shi Wei , Yang Eun Sung , Zhang Yi , Todd John-Paul M. , Wang Lingshu , Andersen Hanne , Foulds Kathryn E. , Edwards Darin K. , Mascola John R. , Moore Ian N. , Lewis Mark G. , Carfi Andrea , Montefiori David , Suthar Mehul S. , McDermott Adrian , Sullivan Nancy J. , Roederer Mario , Douek Daniel C. , Graham Barney S. , Seder Robert A.
Published: April 2021

Immune correlates of protection can be used as surrogate endpoints for vaccine efficacy. The nonhuman primate (NHP) model of SARS-CoV-2 infection replicates key features of human infection and may be used to define immune correlates of protection following vaccination. Here, NHP received either no vaccine or doses ranging from 0.3 – 100 μg of mRNA-1273, a mRNA vaccine encoding the prefusion-stabilized SARS-CoV-2 spike (S-2P) protein encapsulated in a lipid nanoparticle. mRNA-1273 vaccination elicited robust circulating and mucosal antibody responses in a dose-dependent manner. Viral replication was significantly reduced in bronchoalveolar lavages and nasal swabs following SARS-CoV-2 challenge in vaccinated animals and was most strongly correlated with levels of anti-S antibody binding and neutralizing activity. Consistent with antibodies being a correlate of protection, passive transfer of vaccine-induced IgG to naïve hamsters was sufficient to mediate protection. Taken together, these data show that mRNA-1273 vaccine-induced humoral immune responses are a mechanistic correlate of protection against SARS-CoV-2 infection in NHP.

One-Sentence Summary

mRNA-1273 vaccine-induced antibody responses are a mechanistic correlate of protection against SARS-CoV-2 infection in NHP.
Phylogenetics of a Rapid, Continental Radiation: Diversification, Biogeography, and Circumscription of the Beardtongues (Penstemon; Plantaginaceae)
Authors: Wolfe Andrea D. , Blischak Paul D. , Kubatko Laura S.
Published: April 2021

Penstemon(Plantaginaceae), the largest genus of plants native to North America, represents a recent continental evolutionary radiation. We investigated patterns of diversification, phylogenetic relationships, and biogeography, and determined the age of the lineage using 43 nuclear gene loci. We also assessed the current taxonomic circumscription of the ca. 285 species by developing a phylogenetic taxonomic bootstrap method.Penstemonoriginated during the Pliocene/Pleistocene transition. Patterns of diversification and biogeography are associated with glaciation cycles during the Pleistocene, with the bulk of diversification occurring from 1.0–0.5 mya. The radiation across the North American continent tracks the advance and retreat of major and minor glaciation cycles during the past 2.5 million years with founder-event speciation contributing the most to diversification ofPenstemon. Our taxonomic bootstrap analyses suggest the current circumscription of the genus is in need of revision. We propose rearrangement of subgenera, sections, and subsections based on our phylogenetic results. Given the young age and broad distribution ofPenstemonacross North America, it offers an excellent system for studying a rapid evolutionary radiation in a continental setting.
A non-canonical planar cell polarity pathway triggered by light
Authors: Housset Michael , Filion Dominic , Cortes Nelson , Vali Hojatollah , Mandato Craig , Casanova Christian , Cayouette Michel
Published: April 2021

The coordinated spatial arrangement of organelles within a tissue plane, generally referred to as Planar Cell Polarity (PCP), is critical for various processes ranging from organogenesis to sensory detection. Mechanistically, planar cell polarity is typically set-up by gradients of morphogens and their receptors, but whether non-molecular cues, akin to phototropism in plants, might play a part in setting up PCP in animals remains unknown. Here we report that the basal body of newborn photoreceptor cells in the mouse retina is centrally positioned, but then moves laterally around the first post-natal week, thereby generating cell-intrinsic asymmetry. During the second postnatal week, when eyes open, the basal bodies of cone photoreceptor cilia, but not rods, become coordinated across the tissue plane to face the center of the retina. Mechanistically, we show that light is required during a critical window in development, triggering a non-canonical cascade whereby cone transducin interacts with the G- protein signaling modulator protein 2 (GPSM2), which is in turn necessary for cone PCP. This work uncovers a non-canonical PCP pathway initiated by light.
Expanding the genotypic spectrum of ACTG2-related visceral myopathy
Authors: James Kiely N. , Lau Megan , Shayan Katayoon , Lenberg Jerica , Mardach Rebecca , Ignacio Romeo , Halbach Jonathan , Choi Lillian , Kumar Soma , Ellsworth Katarzyna A.
Published: April 2021

Visceral myopathies (VMs) encompass a spectrum of disorders characterized by chronic disruption of gastrointestinal function, with or without urinary system involvement. Pathogenic missense variation in smooth muscle γ-actin gene (ACTG2) is associated with autosomal dominant VM. Whole-genome sequencing of an infant presenting with chronic intestinal pseudo-obstruction revealed a homozygous 187 bp (c.589_613 + 163del188) deletion spanning the exon 6–intron 6 boundary within ACTG2. The patient's clinical course was marked by prolonged hospitalizations, multiple surgeries, and intermittent total parenteral nutrition dependence. This case supports the emerging understanding of allelic heterogeneity in ACTG2-related VM, in which both biallelic and monoallelic variants in ACTG2 are associated with gastrointestinal dysfunction of similar severity and overlapped clinical presentation. Moreover, it illustrates the clinical utility of rapid whole-genome sequencing, which can comprehensively and precisely detect different types of genomic variants including small deletions, leading to guidance of clinical care decisions.
Labour and the Patriot Game
Authors: Hayhurst Mark
Published: April 2021

The Labour Party has recently been advised to showcase its patriotism and let the electorate know it is proud to be British. This is never bad advice for a mainstream political party with designs on power and certainly Labour in the past has enjoyed electoral success when its redistributionist policies have sprung, at least in part, from an obvious love of country. There's a certain urgency, too, in the suggestion, since the Corbyn leadership convinced many former Labour voters that the party was careless about a British identity and perhaps even hostile to British interests. Nevertheless, it would be foolish for Keir Starmer's Labour Party to enter into a destructive competition with the Tories to see who looks better with the national flag. For reasons which the article explores, there are strict limits beyond which a social democratic party would not care to go in such a contest. But this ought not matter. Voters do not want to see Labour leaders telling them how patriotic they are. They are content to be shown. This article surveys the history and looks at some possibilities inherent in Labour and the patriot game.
Development of mycoinsecticide formulations with Beauveria bassiana and Metarhizium brunneum for the control of Orosanga japonica (Hemiptera: Ricaniidae)
Authors: Biryol Seda , Güney Ebru , Eski Ardahan , Bayramoğlu Zeynep , Sezen Kazım , Demirbag Zihni , Demir İsmail
Published: April 2021

Entomopathogenic fungi are promising microbial agents for agricultural pests, especially piercing‐sucking insects. In this study, eight indigenous fungal isolates including four isolates of Metarhizium brunneum and four isolates of Beauveria bassiana were tested on Orosanga japonica Melichar (Hemiptera: Ricaniidae) in order to find an effective control agent that can be utilised for the development of oil‐based fungal mycoinsecticides. In the screening test using 1 × 107 spores ml−1 concentration, KTU‐24 (B. bassiana) and KTU‐51 (M. brunneum) had the highest lethal activity against nymphs and adults of the target pest. Also, at the highest concentration (109 spores ml−1), KTU‐24 showed 92.33 and 94.88% mortality on nymphs and adults at the dose–response tests, respectively, and the KTU‐51 isolate showed 100% mortality on both stages of O. japonica under laboratory conditions. With these results, KTU‐24 and KTU‐51 isolates were used to develop biopesticides. While the mass production of B. bassiana was carried out with liquid‐state fermentation, solid‐state fermentation was used for mass production of M. brunneum. Spores of both isolates were formulated in oil, and products were named as RICANICIDAL Bbas‐TR61 and RICANICIDAL Met‐TR61, respectively. Formulations caused over 97% mortality against nymphs and adults of O. japonica under field conditions and both formulations showed higher efficacy than commercial mycoinsecticides at 108 spores/ml concentration 20 days after treatment. The currents study shows that both mycoinsecticide formulations are highly promising for management of O. japonica.