bluuebunny/crossref_metadata_2025 huggingface dataset:
Multiple structural states of Ca2+-regulated PET hydrolase, Cut190, and its correlation with activity and stability
Authors: Senga Akane , Numoto Nobutaka , Yamashita Mitsuaki , Iida Akira , Ito Nobutoshi , Kawai Fusako , Oda Masayuki
Published: August 2020

An enzyme, Cut190, from a thermophilic isolate, Saccharomonospora viridis AHK190 could depolymerize polyethylene terephthalate (PET). The catalytic activity and stability of Cut190 and its S226P/R228S mutant, Cut190*, are regulated by Ca2+ binding. We previously determined the crystal structures of the inactive mutant of Cut190*, Cut190*S176A, in complex with metal ions, Ca2+ and Zn2+, and substrates, monoethyl succinate and monoethyl adipate. In this study, we determined the crystal structures of another mutant of Cut190*, Cut190**, in which the three C-terminal residues of Cut190* are deleted, and the inactive mutant, Cut190**S176A, in complex with metal ions. In addition to the previously observed closed, open and engaged forms, we determined the ejecting form, which would allow the product to irreversibly dissociate, followed by proceeding to the next cycle of reaction. These multiple forms would be stable or sub-stable states of Cut190, regulated by Ca2+ binding, and would be closely correlated with the enzyme function. Upon the deletion of the C-terminal residues, we found that the thermal stability increased while retaining the activity. The increased stability could be applied for the protein engineering of Cut190 for PET depolymerization as it requires the reaction above the glass transition temperature of PET.
A logic of blockchain updates
Authors: Brünnler Kai , Flumini Dandolo , Studer Thomas
Published: August 2020

Blockchains are distributed data structures that are used to achieve consensus in systems for cryptocurrencies (like Bitcoin) or smart contracts (like Ethereum). Although blockchains gained a lot of popularity recently, there are only few logic-based models for blockchains available. We introduce $\mathsf{BCL}$, a dynamic logic to reason about blockchain updates, and show that $\mathsf{BCL}$ is sound and complete with respect to a simple blockchain model.
Synthesis, chirality-dependent conformational and biological properties of siRNAs containing 5′-(R)- and 5′-(S)-C-methyl-guanosine
Authors: Mikami Atsushi , Erande Namrata , Matsuda Shigeo , Kel’in Alexander , Woods Lauren Blair , Chickering Tyler , Pallan Pradeep S , Schlegel Mark K , Zlatev Ivan , Egli Martin , Manoharan Muthiah
Published: August 2020

Various chemical modifications have been identified that enhance potency of small interfering RNAs (siRNAs) and that reduce off-target effects, immune stimulation, and toxicities of metabolites of these therapeutic agents. We previously described 5′-C-methyl pyrimidine nucleotides also modified at the 2′ position of the sugar. Here, we describe the synthesis of 2′-position unmodified 5′-(R)- and 5′-(S)-C-methyl guanosine and evaluation of these nucleotides in the context of siRNA. The (R) isomer provided protection from 5′ exonuclease and the (S) isomer provided protection from 3′ exonuclease in the context of a terminally modified oligonucleotide. siRNA potency was maintained when these modifications were incorporated at the tested positions of sense and antisense strands. Moreover, the corresponding 5′ triphosphates were not substrates for mitochondrial DNA polymerase. Models generated based on crystal structures of 5′ and 3′ exonuclease oligonucleotide complexes with 5′-(R)- and 5′-(S)-C-methyl substituents attached to the 5′- and 3′-terminal nucleotides, respectively, provided insight into the origins of the observed protections. Structural properties of 5′-(R)-C-methyl guanosine incorporated into an RNA octamer were analysed by X-ray crystallography, and the structure explains the loss in duplex thermal stability for the (R) isomer compared with the (S) isomer. Finally, the effect of 5′-C-methylation on endoribonuclease activity has been explained.
NEO100 enables brain delivery of blood‒brain barrier impermeable therapeutics
Authors: Wang Weijun , Marín-Ramos Nagore I , He Haiping , Zeng Shan , Cho Hee-Yeon , Swenson Stephen D , Zheng Long , Epstein Alan L , Schönthal Axel H , Hofman Florence M , Chen Ligang , Chen Thomas C
Published: August 2020

Background

Intracarotid injection of mannitol has been applied for decades to support brain entry of therapeutics that otherwise do not effectively cross the blood–brain barrier (BBB). However, the elaborate and high-risk nature of this procedure has kept its use restricted to well-equipped medical centers. We are developing a more straightforward approach to safely open the BBB, based on the intra-arterial (IA) injection of NEO100, a highly purified version of the natural monoterpene perillyl alcohol.

Methods

In vitro barrier permeability with NEO100 was evaluated by transepithelial/transendothelial electrical resistance and antibody diffusion assays. Its mechanism of action was studied by western blot, microarray analysis, and electron microscopy. In mouse models, we performed ultrasound-guided intracardiac administration of NEO100, followed by intravenous application of Evan’s blue, methotrexate, checkpoint-inhibitory antibodies, or chimeric antigen receptor (CAR) T cells.

Results

NEO100 opened the BBB in a reversible and nontoxic fashion in vitro and in vivo. It enabled greatly increased brain entry of all tested therapeutics and was well tolerated by animals. Mechanistic studies revealed effects of NEO100 on different BBB transport pathways, along with translocation of tight junction proteins from the membrane to the cytoplasm in brain endothelial cells.

Conclusion

We envision that this procedure can be translated to patients in the form of transfemoral arterial catheterization and cannulation to the cerebral arteries, which represents a low-risk procedure commonly used in a variety of clinical settings. Combined with NEO100, it is expected to provide a safe, widely available approach to enhance brain entry of any therapeutic.
Depression Partially Mediates the Association of Adverse Childhood Experiences with Pain Intensity in Patients with Chronic Pelvic Pain Syndrome: Results from a Cross-Sectional Patient Survey
Authors: Piontek Katharina , Apfelbacher Christian , Ketels Gesche , Brünahl Christian , Löwe Bernd
Published: August 2020

Objective

Adverse childhood experiences (ACEs), such as emotional and physical maltreatment, are linked to chronic pelvic pain syndrome (CPPS) in adults. Psychological factors are important in understanding CPPS. We aimed to determine the nature and frequency of ACEs in male and female patients with CPPS and to investigate whether somatic symptoms and psychological comorbidities mediate the relationship of ACE severity with pain intensity.

Design

Cross-sectional study.

Setting

Interdisciplinary outpatient clinic for CPPS in Hamburg, Germany.

Subjects

Individuals with CPPS (n = 234) who were 18 to 84 years of age.

Methods

Using a self-administered questionnaire, we assessed the history of ACEs (ACE Scale), pain intensity (McGill Pain Questionnaire), somatic symptoms (Patient Health Questionnaire–15]), depression (Patient Health Questionnaire–9), and anxiety (Generalized Anxiety Disorder Scale). Parallel mediation analysis was conducted to examine whether the association of ACE severity with pain intensity is mediated by somatic symptoms, depression, and anxiety.

Results

Emotional abuse and neglect were reported more than twice as often as physical abuse and neglect (37.2% vs 17.1%). Depression partially mediated the association of ACE severity with pain intensity in the whole study population. In sex-stratified analyses, different patterns of associations were observed, but somatic symptoms predicted pain intensity in both sexes.

Conclusions

Emotional maltreatment was highly prevalent, supporting an increased consideration of psychological factors in CPPS and indicating the need to screen for ACEs in patients with CPPS. Findings further suggest that depression and somatic symptoms may be important targets for therapeutic interventions in patients with CPPS who have a history of childhood adversity.
Peripheral Nerve Stimulation: A New Treatment for Meralgia Paresthetica
Authors: Langford Brendan , Mauck William D
Published: August 2020

Background

Meralgia paresthetica is a condition caused by entrapment of the lateral femoral cutaneous nerve that leads to paresthesia along the anterolateral portion of the thigh. Because of advancements in neuromodulation, peripheral nerve stimulation (PNS) has been considered a new treatment option for meralgia paresthetica. Newer PNS technology targets peripheral nerves directly yet in a minimally invasive manner. We report a case in which a PNS device provided more than 12 months of complete pain relief in a patient with meralgia paresthetica and helped the patient avoid a neurolysis procedure.

Case Presentation

A 57-year-old male presented to clinic with a 6-year history of “painful numbness [and] burning” along the right lateral thigh. He rated his pain as 8 out of 10, which decreased to a rating of 2 out of 10 with the use of gabapentin, but unwanted side effects motivated him to seek alternative treatment. On the basis of his history, physical exam, and imaging results, he was diagnosed with meralgia paresthetica. He was offered neurolysis; however, after seeing a pain specialist, he agreed to the implantation of a SPRINT peripheral nerve stimulator. After the implantation procedure, his pain reduced to 0 out of 10, and his quality of life improved, with better sleep and less somnolence. The device was removed after 60 days, as planned. He continued to have complete resolution of pain at 12 months after the date of device implantation.

Conclusion

With recent advancements, PNS can be used to treat meralgia paresthetica in an effective yet minimally invasive manner. As newer PNS technology becomes more familiar to physicians and pain specialists, it is likely to be used as a mainstay treatment for meralgia paresthetica.
The Yield Spread and Bond Return Predictability in Expansions and Recessions
Authors: Andreasen Martin M , Engsted Tom , Møller Stig V , Sander Magnus
Published: August 2020

This paper uncovers that expected excess bond returns display a positive correlation with the slope of the yield curve (i.e., yield spread) in expansions but a negative correlation in recessions. We use a macro-finance term structure model with different market prices of risk in expansions and recessions to show that a very accommodating monetary policy in recessions is a key driver of this switch in return predictability.
Immunotoxicity of an Electrochemically Fluorinated Aqueous Film-Forming Foam
Authors: McDonough Carrie A , Ward Chastity , Hu Qing , Vance Samuel , Higgins Christopher P , DeWitt Jamie C
Published: August 2020

Aqueous film-forming foams (AFFFs) are complex per- and polyfluoroalkyl substance (PFAS)-containing mixtures used extensively as fire suppressants. AFFF-impacted groundwater and surface water have contaminated drinking water with PFASs in many communities, raising concerns about health effects from drinking water exposures. As individual PFASs have been identified as immune hazards, the immunotoxicity of complex PFAS mixtures is also a concern. Adult female and male C57BL/6 mice were given a commercial AFFF formulation for 10 days via gavage; administered dose was based on combined content of perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS) measured in the formulation (0, 1.88, 3.75, 7.5, or 10 mg PFOS+PFOA/kg body weight). A PFOA positive control of 7.5 mg/kg body weight was also given. Compared with the 0 mg/kg group, the following changes were noted: Body weights of males exposed to 7.5 and 10 mg PFOS+PFOA/kg were reduced by 15%, on average; female body weights did not differ. Average relative liver weights were increased 50%–200% in males and 37.5%–193% in females and liver peroxisome proliferation was increased 2- to 12-fold in all doses of both sexes. Antigen-specific antibody production was suppressed, on average, by 13% in males and by 12.4% in females across all doses. Spleen cellularity and lymphocyte subpopulations did not differ by dose for either sex. Our data indicate that though this complex PFAS mixture contained fairly low PFOA content, it induced changes in C57BL/6 mice similar to changes induced by PFOA alone, likely due to the presence of PFOS and many other PFASs.
A Survey of Neonatal Clinicians' Use, Needs, and Preferences for Kangaroo Care Devices
Authors: Weber Ashley , Jackson Yamile
Published: August 2020

Background:

Decades of research supports the benefits of kangaroo care (KC) for the parent and newborn. Supportive KC devices may be an important tool clinicians can use to assist parents with KC. In recent years, there has been a rise in the availability of KC devices. However, the use, needs, and preferences for these supportive devices by neonatal clinicians have not been documented.

Purpose:

To survey clinicians' use, needs, and preferences of KC supportive devices, and examine whether differences exist based on clinician and organizational characteristics.

Methods:

A cross-sectional, online survey was sent through neonatal organization Web sites, conferences, and social media.

Results:

Many clinicians (n = 68, 43%; N = 158) facilitated KC with a supportive device, with 81% of devices provided by the clinician's employer. The most important “Must Have” feature of a KC device was “Safety: Reduces patient falls if caregiver sleeps or needs to use hands” (84% of respondents) followed by washability (82%), and “immediate, effective access to the baby” (78%). Clinicians' responses did not differ based on hospital setting, type of unit, KC experience, or experience using a KC device.

Implications for Practice:

To support safe use of KC devices in neonatal intensive care unit (NICU) clinical care, a device must hold the proper KC position consistently, allow immediate access to the infant, and hold the infant in place without the parent's hands to prevent falls. Training is needed to ensure safe device use.

Implications for Research:

Future research should evaluate the safety, efficacy, and cost-effectiveness of these devices.
Opioid Prescribing Patterns for Low Back Pain Among Commercially Insured Children
Authors: Azad Tej D. , Harries Michael D. , Veeravagu Anand , Ratliff John K.
Published: August 2020

Level of Evidence: 3
Age at menarche in Seveso daughters exposed in utero to 2,3,7,8-tetrachlorodibenzo-p-dioxin
Authors: Warner Marcella , Rauch Stephen , Ames Jennifer , Mocarelli Paolo , Brambilla Paolo , Eskenazi Brenda
Published: August 2020

Background:

In utero exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is associated with delayed pubertal development in animal studies. No epidemiologic study has investigated this association. We examined the relationship of in utero exposure to TCDD with reported age at onset of menarche in female children born to a unique cohort of TCDD-exposed women resulting from an explosion in Seveso, Italy, on 10 July 1976.

Methods:

In 2014, nearly 40 years after the explosion, we enrolled postexplosion offspring, 2 to 39 years of age, in the Seveso Second Generation Study. Age at onset of menarche (years) was collected for 316 daughters by maternal report or self-report at interview. In utero TCDD exposure was defined by maternal TCDD serum concentrations extrapolated to the pregnancy.

Results:

At interview, 287 daughters were postmenarche and reported age at menarche averaged 12.1 years (±1.3 years). Overall, we found no change in risk of menarche onset with a 10-fold increase in in utero TCDD exposure (hazard ratio [HR] = 0.86; 95% confidence interval [CI] = 0.71, 1.04). When we considered maternal menarche status in 1976 as a potentially sensitive developmental exposure window, in utero TCDD (log10) was associated with later age at menarche among daughters whose mothers were premenarche (HR = 0.71; 95% CI = 0.52, 0.97) but not postmenarche (HR = 0.89; 95% CI = 0.71, 1.12) at the time of the explosion (P int = 0.24).

Conclusions:

These results suggest that in utero TCDD exposure may alter pubertal timing among daughters of women who were prepubescent at the time of the Seveso accident.
CHARACTERISTICS AND SURGICAL OUTCOMES OF RHEGMATOGENOUS RETINAL DETACHMENT IN OLDER ADULTS
Authors: Patel Samir N. , Starr Matthew R. , Obeid Anthony , Ryan Edwin H. , Ryan Claire , Forbes Nora J. , Soares Rebecca R. , Ammar Michael , Patel Luv G. , Capone Antonio , Emerson Geoffrey G. , Joseph Daniel P. , Eliott Dean , Regillo Carl D. , Gupta Omesh P. , Hsu Jason , Yonekawa Yoshihiro ,   for the Primary Retinal Detachment Outcomes (PRO) Study Group
Published: August 2020

Purpose:

To describe characteristics and outcomes of primary rhegmatogenous retinal detachment in older adults (age ≥ 80).

Methods:

Consecutive patients with rhegmatogenous retinal detachment undergoing pars plana vitrectomy (PPV), scleral buckling (SB), or PPV/SB in the Primary Retinal Detachment Outcomes Study were evaluated. Outcome measures included single surgery anatomic success and visual acuity.

Results:

Of 2,144 patients included, 125 (6%) were 80 years or older. Compared with younger patients (age 40–79), older adults were more likely to be pseudophakic (P < 0.001), have macula-off detachments (P < 0.001), and have preoperative proliferative vitreoretinopathy (P = 0.02). In older adults, initial surgery was PPV in 73%, PPV/SB in 27%, and primary SB in 0%. Single surgery anatomic success was 78% in older adults compared with 84% in younger patients (P = 0.03). In older adults, single surgery anatomic success was 74% for PPV and 91% for PPV/SB (P = 0.03). The final mean logMAR was lower for older adults (0.79 [20/125] vs. 0.40 [20/40], [P < 0.001]). In older adults, the final mean logMAR for eyes that underwent PPV was 0.88 (20/160) compared with 0.50 (20/63) for PPV/SB (P = 0.03).

Conclusion:

Octogenarians and nonagenarians presented with relatively complex pseudophakic rhegmatogenous retinal detachments. Single surgery anatomic success and visual outcomes were worse compared with younger patients, and PPV/SB had better outcomes compared with PPV alone.
A Cluster of Corneal Donor Rim Cultures Positive for Achromobacter Species Associated With Contaminated Eye Solution
Authors: André Camille , Durand Marlene L. , Buckley Thomas , Cadorette James , Gilmore Michael S. , Ciolino Joseph B. , Bispo Paulo J. M.
Published: August 2020

Purpose:

To investigate a cluster of corneoscleral rim cultures positive for Achromobacter species over a 6-month period at Massachusetts Eye and Ear.

Methods:

An increased rate of positive corneal donor rim cultures was noted at Massachusetts Eye and Ear between July and December 2017. Positive cultures were subjected to identification and antimicrobial susceptibility testing by phenotypic (MicroScan WalkAway) and genotypic (16S rDNA sequencing) methods. Samples of the eye wash solution (GeriCare) used in the eye bank were also evaluated. Antimicrobial activity of Optical-GS against Achromobacter spp. at 4°C and 37°C was assessed by time-kill kinetics assay.

Results:

Of 99 donor rims cultured, 14 (14.1%) grew bacteria with 11 (78.6%) due to uncommon nonfermenting Gram-negative bacilli. These had been identified by standard automated methods as Achromobacter (n = 3), Alcaligenes (n = 3), Ralstonia (n = 2), Pseudomonas (n = 2), and Stenotrophomonas (n = 1). Eight of these 11 isolates were subsequently available for molecular identification, and all were identified as Achromobacter spp. Six bottles of eyewash solution were evaluated and were positive for abundant Achromobacter spp. (3.4 × 105 ± 1.1 CFU/mL). Optisol-GS had no bactericidal activity against Achromobacter spp. at 4°C after 24-hour incubation but was bactericidal at 37°C. None of the patients who had received the contaminated corneas developed postoperative infection.

Conclusions:

An eyewash solution arising from a single lot was implicated in the contamination of donor rims by Achromobacter spp. The isolates were able to survive in the Optisol-GS medium at the recommended storage temperature. This highlights the need to continue improving protocols for tissue preparation and storage.
Combined Epithelial, Stromal, and Anterior Chamber Ingrowth After Penetrating Keratoplasty
Authors: Farahani Mina , Minckler Don , Chu Joshua , Huynh Kyle , Farid Marjan
Published: August 2020

Purpose:

To present a case of primary graft failure after penetrating keratoplasty found to have epithelial ingrowth into the host stroma on histopathologic analysis.

Methods:

This is a single observational case report.

Results:

We herein describe the clinical course of a case of primary graft failure after penetrating keratoplasty. The corneal button was sent for histopathologic analysis. Analysis of the patient's failed corneal button revealed circumferential epithelial full-thickness wound invasion and stromal epithelial invasion into corneal stroma.

Conclusions:

Based on histopathologic analysis and this patient's presentation, the stromal ingrowth followed recipient epithelial invasion of the wound and stromal invasion through clefts in the donor corneal edges. Cases of primary graft failure should be assessed for histopathologic evidence of epithelial stromal ingrowth, despite its rarity. To our knowledge, epithelial ingrowth into the corneal donor stroma after penetrating keratoplasty has not been previously reported.
Type D Personality Associated With Increased Risk for Mortality in Adults With Congenital Heart Disease
Authors: Kauw Dirkjan , Schoormans Dounya , Sieswerda Gertjan Tj. , Van Melle Joost P. , Vliegen Hubert W. , Van Dijk Arie P.J. , Hulsbergen-Zwarts Mariët S. , Post Marco C. , Ansink Tieneke J. , Mulder Barbara J.M. , Bouma Berto J. , Schuuring Mark J.
Published: August 2020

Background

Type D personality has been previously shown to increase the risk for mortality in patients with acquired heart disease.

Objective

We aimed to compare mortality in adult patients with congenital heart disease (CHD) with and without type D.

Methods

Survival was assessed using prospective data from the Dutch national Congenital Corvitia registry for adults with CHD. Patients were randomly selected from the registry and characterized at inclusion in 2009 for the presence of type D using the DS14 questionnaire.

Results

One thousand fifty-five patients, with 484 (46%) males, a mean (SD) age of 41 (14) years, 613 (58%) having mild CHD, 348 (33%) having moderate CHD, and 94 (9%) having severe CHD, were included. Type D personality was present in 225 patients (21%). Type D was associated with an increased risk for all-cause mortality independent of age, sex, New York Heart Association class, number of prescribed medications, depression, employment status, and marital status (hazard ratio, 1.94; 95% confidence interval, 1.05–3.57; P = .033).

Conclusion

Type D personality was associated with an increased risk for all-cause mortality in adult patients with CHD.
Improving Timeliness of Internal Medicine Consults in the Emergency Department: A Quality Improvement Initiative
Authors: Beckerleg Weiwei , Hasimja-Saraqini Delvina , Kwok Edmund S. H. , Hamdy Noha , Battram Erica , Wooller Krista R.
Published: August 2020

Introduction:

Emergency department (ED) wait time is an important health system quality indicator. Prolonged consult to decision time (CTDT), the time it takes to reach a disposition decision after receiving a specialty consultation request, can contribute to increased overall length of stay in the ED.

Objective:

To identify delays in the consultation process for general internal medicine (GIM) and trial interventions to reduce CTDT.

Methods:

The study was conducted at a large tertiary teaching hospital with GIM inpatient wards at two campuses. Four interventions were trialed over sequential Plan-Do-Study-Act cycles: (1) process mapping, (2) resident education sessions, (3) audit and feedback of CTDT, and (4) adding a swing shift during peak consult volume.

Measurements:

The primary outcome measures were mean CTDT for patients admitted to GIM and the proportion of admitted patients with CTDT of less than 3 hours.

Results:

Mean CTDT decreased from 4.61 hours before intervention to 4.18 hours after intervention (p < .0001). The proportion of GIM patients with CTDT less than 3 hours increased from 25% to 33% (p < .0001).

Conclusions:

The interventions trialed led to a sustained reduction in CTDT over a 12-month period and demonstrated the effectiveness of education in influencing physician performance.
Effect of blood glucose and body weight on image quality in brain [18F]FDG PET imaging
Authors: Henriksen Otto M. , Holm Søren , Marner Lisbeth , Law Ian
Published: August 2020

Objectives

The aims of the present study were to assess the influence of mild to moderate hyperglycaemia and body weight on brain 2-[18F]fluoro-2-deoxy-d-glucose ([18F]FDG) PET, and to what extent a simple algorithm for maintaining count density may compensate for these effects.

Methods

We prospectively included 63 patients undergoing routine brain [18F]FDG PET. Scan time and injected activity were adjusted in patients with hyperglycaemia or increased body weight. Measures of perceived image quality, image noise and image contrast were obtained in both standard scans and intervention scans.

Results

Elevated blood glucose and increased body weight were associated with reduced count density and increased image noise that in turn were associated with lower scores of perceived image quality. The proposed simple algorithm effectively maintained the image noise level and improved perceived image quality across the full range of elevated blood glucose values and body weights, although the effect of intervention on perceived image quality was attenuated by lower image contrast in patients with moderate hyperglycaemia. In patients with increased body weight or blood glucose, the fraction of scans with poor image quality decreased from 9/29 to 2/29 (P = 0.04) and the fraction with good image quality increased from 7/29 to 20/29 (P = 0.001) when applying the proposed algorithm.

Conclusions

Increasing blood glucose and body weight are associated with increased image noise in standard imaging conditions. Improving count density by prolonging scan time and increasing injected activity significantly improves image quality in hyperglycaemic patients, although the image contrast remains reduced in patients with most pronounced hyperglycaemia.
Understanding Variation in Postacute Care
Authors: Reistetter Timothy A. , Eschbach Karl , Prochaska John , Jupiter Daniel C. , Hong Ickpyo , Haas Allen M. , Ottenbacher Kenneth J.
Published: August 2020

Objective

The aims of the study were to demonstrate a method for developing rehabilitation service areas and to compare service areas based on postacute care rehabilitation admissions to service areas based on acute care hospital admissions.

Design

We conducted a secondary analysis of 2013–2014 Medicare records for older patients in Texas (N = 469,172). Our analysis included admission records for inpatient rehabilitation facilities, skilled nursing facilities, long-term care hospitals, and home health agencies. We used Ward’s algorithm to cluster patient ZIP Code Tabulation Areas based on which facilities patients were admitted to for rehabilitation. For comparison, we set the number of rehabilitation clusters to 22 to allow for comparison to the 22 hospital referral regions in Texas. Two methods were used to evaluate rehabilitation service areas: intraclass correlation coefficient and variance in the number of rehabilitation beds across areas.

Results

Rehabilitation service areas had a higher intraclass correlation coefficient (0.081 vs. 0.076) and variance in beds (27.8 vs. 21.4). Our findings suggest that service areas based on rehabilitation admissions capture has more variation than those based on acute hospital admissions.

Conclusions

This study suggests that the use of rehabilitation service areas would lead to more accurate assessments of rehabilitation geographic variations and their use in understanding rehabilitation outcomes.
A New Surgical Method for Medial Ectropion Repair: Fixing the Lower Eyelid Retractor to the Back of the Tarsal Plate
Authors: Kokubo Kenichi , Katori Nobutada , Hayashi Kengo , Fujii Akiko , Kitamura Sho , Maeda Chie , Haga Shoko , Kawabata Tomotaka , Maegawa Jiro
Published: August 2020

Involutional ectropion is a disease in which the eyelids are everted outwards, and because the eyelids move away from the eyeballs, the ocular surface and conjunctiva are exposed causing inflammation, pain, photophobia, foreign body sensation, epiphora, and blurred vision. It is thought to be caused by horizontal and vertical laxity. Various surgical methods have reportedly been used to correct involutional ectropion. Shortening the lower eyelid retractor (LER) is an indispensable surgical operation for medial ectropion. When the LER is shortened, it is usually fixed to the lower edge of the tarsal plate. Herein we describe a new type of surgery that has now been performed on 6 eyes in 4 patients. The procedure involves separating the conjunctiva from the tarsal plate, inserting the LER between the conjunctiva and the tarsal plate, and then fixing it to the back of the tarsal plate. In all 6 eyes, the lower eyelid now contacts the eyeball, and morphological improvements were achieved. This new surgical method is a useful way to raise the tarsal plate.
Three-Dimensional Analysis of Craniofacial Structures of Individuals With Nonsyndromic Unilateral Complete Cleft Lip and Palate
Authors: Parveen Shahistha , Husain Akhter , Johns Gejo , Mascarenhas Rohan , Reddy Srinivas Gosla
Published: August 2020

Cleft lip and palate (CLP) is one of the most common congenital deformities. Primary surgeries at an early age result in scar formation, which may impede the growth of craniofacial structures of the maxilla. Orthodontist's role in the management of individuals with CLP is important and starts from the time of birth. The knowledge of craniofacial structures in individuals with a cleft is essential for treatment planning. The purpose of this study was to analyze and compare craniofacial structures of cleft and noncleft side of individuals with non-syndromic unilateral complete cleft lip and palate (NSUCCLP) using cone-beam computed tomography (CBCT). CBCT scans of individuals with NSUCCLP (n = 42) were retrieved from the databases of two cleft centers, which followed the same protocols for timing and type of primary surgeries and secondary alveolar bone grafting (SABG). DICOM files of CBCT scans were integrated into Dolphin 3D software, and analysis was carried out in multiplanar views. The craniofacial structures of individuals with NSUCCLP were analyzed using fourteen parameters. Measurements were also recorded between the cleft and noncleft sides for comparison. The volume of the maxilla was generated by isolating it from adjacent structures on a 3D reconstructed model. MAWC, MAWPM1, MAWPM2, MAWM1, and MV of the cleft side was less than noncleft side (P < 0.05). MHP @ N Aper is less on the noncleft side (P < 0.05). There is an asymmetry of structures around the dentoalveolar and nasal region; however, asymmetries were not affected at deeper structures of the craniofacial region of individuals with NSUCCLP.
Identification of specific amino acid residues in the border disease virus glycoprotein E2 that modify virus growth in pig cells but not in sheep cells
Authors: Fahnøe Ulrik , Deng Yu , Davids Nana A. , Lohse Louise , Bukh Jens , Belsham Graham J. , Rasmussen Thomas Bruun
Published: August 2020

Border disease virus (BDV) envelope glycoprotein E2 is required for entry into cells and is a determinant of host tropism for sheep and pig cells. Here, we describe adaptive changes in the BDV E2 protein that modify virus replication in pig cells. To achieve this, two BDV isolates, initially collected from a pig and a sheep on the same farm, were passaged in primary sheep and pig cells in parallel with a rescued variant of the pig virus derived from a cloned full-length BDV cDNA. The pig isolate and the rescued virus shared the same amino acid sequence, but the sheep isolate differed at ten residues, including two substitutions in E2 (K771E and Y925H). During serial passage in cells, the viruses displayed clear selectivity for growth in sheep cells; only the cDNA-derived virus adapted to grow in pig cells. Sequencing revealed an amino acid substitution (Q739R) in the E2 domain DA of this rescued virus. Adaptation at the same residue (Q739K/Q739R) was also observed after passaging of the pig isolate in sheep cells. Use of reverse genetics confirmed that changing residue Q739 to R or K (each positively charged) was sufficient to achieve adaptation to pig cells. Furthermore, this change in host tropism was suppressed if Q739R was combined with K771E. Another substitution (Q728R), conferring an additional positive charge, acquired during passaging, restored the growth of the Q739R/K771E variant. Overall, this study provided evidence that specific, positively charged, residues in the E2 domain DA are crucial for pig-cell tropism of BDV.
Genetic and epigenetic contributions to variation in transposable element expression responses to abiotic stress in maize
Authors: Liang Zhikai , Anderson Sarah N. , Noshay Jaclyn M. , Crisp Peter A. , Enders Tara A. , Springer Nathan M.
Published: August 2020

Transposable elements (TEs) pervade most eukaryotic genomes but the repetitive nature of TEs has complicated the analysis of their expression. Although the majority of TEs are silent, we document the activation of some TEs during abiotic stress.

TE expression was monitored in seedling leaf tissue of maize inbreds subjected to heat or cold stress conditions. DNA methylation profiles and comparative genomics were used to probe the variability of TE expression responses.

Although there was no evidence for a genome-wide activation of TEs, a subset of TE families generate transcripts only in stress conditions. There is substantial variation for which TE families exhibit stress-responsive expression in the three genotypes. The stress-responsive activation of a TE family can often be attributed to a small number of elements in the family. These elements that are activated often contain small regions lacking DNA methylation, while fully methylated elements are rarely expressed. A comparison of the expression of specific TEs in different maize genotypes reveals high levels of variability that can be attributed to both genome content differences and epigenetic variation.

This study provides insights into the genetic and epigenetic factors that influence TE regulation in normal and stress conditions.
Serum Amyloid P inhibits single stranded RNA-induced lung inflammation, lung damage, and cytokine storm in mice
Authors: Karhadkar Tejas R. , Pilling Darrell , Gomer Richard H.
Published: August 2020

SARS-CoV-2 is a single stranded RNA (ssRNA) virus and contains GU-rich sequences distributed abundantly in the genome. In COVID-19, the infection and immune hyperactivation causes accumulation of inflammatory immune cells, blood clots, and protein aggregates in lung fluid, increased lung alveolar wall thickness, and upregulation of serum cytokine levels. A serum protein called serum amyloid P (SAP) has a calming effect on the innate immune system and shows efficacy as a therapeutic for fibrosis in animal models and clinical trials. In this report, we show that aspiration of the GU-rich ssRNA oligonucleotide ORN06 into mouse lungs induces all of the above COVID-19-like symptoms. Men tend to have more severe COVID-19 symptoms than women, and in the aspirated ORN06 model, male mice tended to have more severe symptoms than female mice. Intraperitoneal injections of SAP starting from day 1 post ORN06 aspiration attenuated the ORN06-induced increase in the number of inflammatory cells and formation of clot-like aggregates in the mouse lung fluid, reduced ORN06-increased alveolar wall thickness and accumulation of exudates in the alveolar airspace, and attenuated an ORN06-induced upregulation of the inflammatory cytokines IL-1β, IL-6, IL-12p70, IL-23, and IL-27 in serum. Together, these results suggest that aspiration of ORN06 is a simple model for both COVID-19 as well as cytokine storm in general, and that SAP is a potential therapeutic for diseases with COVID-19-like symptoms as well as diseases that generate a cytokine storm.
Novel combination of CRISPR-based gene drives eliminates resistance and localises spread
Authors: Faber Nicky R. , McFarlane Gus R. , Gaynor R. Chris , Pocrnic Ivan , Whitelaw C. Bruce A. , Gorjanc Gregor
Published: August 2020

Invasive species are among the major driving forces behind biodiversity loss. Gene drive technology may offer a humane, efficient and cost-effective method of control. For safe and effective deployment it is vital that a gene drive is both self-limiting and can overcome evolutionary resistance. We present HD-ClvR, a novel combination of CRISPR-based gene drives that eliminates resistance and localises spread. As a case study, we model HD-ClvR in the grey squirrel (Sciurus carolinensis), which is an invasive pest in the UK and responsible for both biodiversity and economic losses. HD-ClvR combats resistance allele formation by combining a homing gene drive with a cleave-and-rescue gene drive. The inclusion of a self-limiting daisyfield gene drive allows for controllable localisation based on animal supplementation. We use both randomly mating and spatial models to simulate this strategy. Our findings show that HD-ClvR can effectively control a targeted grey squirrel population, with little risk to other populations. HD-ClvR offers an efficient, self-limiting and controllable gene drive for managing invasive pests.
Common Marmoset Gut Microbiome Profiles in Health and Intestinal Disease
Authors: Sheh Alexander , Artim Stephen C. , Burns Monika A. , Molina-Mora Jose Arturo , Lee Mary Anne , Dzink-Fox JoAnn , Muthupalani Sureshkumar , Fox James G.
Published: August 2020

Chronic gastrointestinal (GI) diseases are the most common diseases in captive marmosets. The gut microbiome of healthy (n=91), inflammatory bowel disease (IBD) (n=59), and duodenal ulcer/stricture (n=23) captive marmosets was characterized. Healthy marmosets exhibited a “humanized,”Bacteroidetes-dominantmicrobiome. Despite standardized conditions, cohorts subdivided intoPrevotella- andBacteroides-dominantgroups based on marmoset source. IBD was highest in aPrevotella-dominant cohort while strictures were highest in aBacteroides-dominant cohort. Stricture-associated dysbiosis was characterized byAnaerobiospirillumloss andClostridium perfringensincreases. Stricture tissue presented upregulation of lipid metabolism genes and increased abundance ofC. perfringens, a causative agent of GI diseases and intestinal strictures in humans. IBD was associated with a lowerBacteroides:P. copriratio within each source. Consistent withPrevotella-linked diseases, pro-inflammatory genes were upregulated. This report highlights the humanization of the captive marmoset microbiome and its potential as a “humanized” animal model ofC. perfringens-inducedenteritis/strictures andP. copri-associatedIBD.
Crucial Role ofSalmonellaGenomic Island 1 Master Activator in Parasitism of IncC plasmids
Authors: Durand Romain , Huguet Kévin T. , Rivard Nicolas , Carraro Nicolas , Rodrigue Sébastien , Burrus Vincent
Published: August 2020

IncC conjugative plasmids and the multiple variants ofSalmonellaGenomic Island 1 (SGI1) are two functionally interacting families of mobile genetic elements commonly associated with multidrug resistance inGammaproteobacteria. SGI1 and its siblings are specifically mobilisedin transby IncC conjugative plasmids. Conjugative transfer of IncC plasmids is activated by the plasmid-encoded master activator AcaCD. SGI1 carries five AcaCD-responsive promoters that drive the expression of genes involved in its excision, replication, and mobilisation. SGI1 encodes an AcaCD homologue, the transcriptional activator complex SgaCD (also known as FlhDCSGI1) that seems to recognise and activate the same SGI1 promoters. Here, we investigated the relevance of SgaCD in SGI1’s lifecycle. Mating assays revealed the requirement for SgaCD and its IncC-encoded counterpart AcaCD in the mobilisation of SGI1. An integrative approach combining ChIP-exo, Cappable-seq, and RNA-seq confirmed that SgaCD activates each of the 18 AcaCD-responsive promoters driving the expression of the plasmid transfer functions. A comprehensive analysis of the activity of the complete set of AcaCD-responsive promoters in both SGI1 and IncC plasmid was performed through reporter assays. qPCR and flow cytometry assays revealed that SgaCD is essential for the excision and replication of SGI1, and the destabilisation of the helper IncC plasmid.
Presence of the Hmq system and production of 4-hydroxy-3-methyl-2-alkylquinolines is heterogeneously distributed betweenBurkholderia cepaciacomplex species and more prevalent among environmental than clinical isolates
Authors: Coulon Pauline M.L. , Zlosnik James E.A. , Déziel Eric
Published: August 2020

SomeBurkholderia cepaciacomplex (Bcc) strains have been reported to produce 4-hydroxy-3-methyl-2-alkylquinolines (HMAQs), analogous to the 4-hydroxy-2-alkylquinolines ofPseudomonas aeruginosa. Usingin silicoanalyses, we previously showed that thehmqABCDEFGoperon, which encodes enzymes involved in the biosynthesis of HMAQs, is carried by about one-third of Bcc strains, with considerable inter- and intra-species variability. In the present study, we investigated by PCR, using consensus primers, the distribution ofhmqABCDEFGin a collection of 313 Bcc strains (222 of clinical and 91 of environmental origins) - belonging to 18 Bcc species. We confirmed that the distribution is species-specific, although not all strains within a species carry thehmqABCDEFGoperon. Among the 30% of strains bearing thehmqABCDEFGoperon, we measured the total HMAQs production and showed that 90% of environmental isolates and 68% of clinically isolated Bcc produce detectable levels of HMAQs when cultured in TSB medium. For the strains having thehmqABCDEFGoperon but not producing HMAQs, we studied the transcription and showed that none expressed thehmqAgene under the specified culture conditions. Interestingly, thehmqABCDEFGoperon is more prevalent among plant root environment species (e.g.B. ambifaria, B. cepacia) and absent in species commonly found in chronically colonized individuals with cystic fibrosis (e.g.B. cenocepacia, B. multivorans), suggesting that the Hmq system could play a role in niche adaptation by influencing rhizosphere microbial community and could have been lost through evolution. Understanding the Hmq system and its regulation will provide clues concerning the production of HMAQs and their functions in Bcc.
First isolation and genetic characterization of Puumala orthohantavirus strains from France
Authors: Vulin Johann , Murri Séverine , Madrières Sarah , Galan Maxime , Tatard Caroline , Piry Sylvain , Vaccari Gabriele , D’agostino Claudia , Charbonnel Nathalie , Castel Guillaume , Marianneau Philippe
Published: August 2020

Puumala orthohantavirus (PUUV) causes a mild form of haemorrhagic fever with renal syndrome (HFRS) called nephropathia epidemica (NE), regularly diagnosed in Europe. France represents the western frontier of the expansion of NE in Europe with two distinct areas: an endemic area (north-eastern France) where PUUV circulates in rodent populations, with detection of many human NE cases, and a non-endemic area (south-western France) where the virus is not detected, with only a few human cases being reported. France is thus a relevant country in which to study the factors that influence the evolution of PUUV distribution. In this study, we describe for the first time the isolation of two PUUV strains from two distinct French geographical areas: Ardennes (endemic area) and Loiret (non-endemic area). To isolate PUUV efficiently, we selected wild bank voles (Myodes glareolus, the specific reservoir of PUUV) captured in these areas and that were seronegative for anti-PUUV IgG (ELISA), but showed a non-negligible viral RNA load in their lung tissue (qRT-PCR). With this study design, we were able to cultivate and maintain these two strains in Vero E6 cells and also propagate both strains in immunologically neutral bank voles efficiently and rapidly. Complete coding sequences of the S and M segments were determined by Sanger sequencing from RNA extracted from positive bank voles (naturally and experimentally infected) and from supernatants of Vero E6 cell extracts. For the M segment, nucleotide sequences were 100% identical for both strains. For the S segment, the amino-acid sequences from each strain revealed one mismatch between sequences obtained from tissue and from cell supernatants, revealing distinct “bank vole” and a “cell” molecular profile. High-throughput sequencing confirmed Sanger results, and provided a better assessment of the impact of isolation methods on intra-host viral diversity.
Evading resistance to gene drives
Authors: Gomulkiewicz Richard , Thies Micki L. , Bull James J.
Published: August 2020

Gene drives offer the possibility of altering and even suppressing wild populations of countless plant and animal species, and CRISPR technology now provides the technical feasibility of engineering them. However, population-suppression gene drives are prone to select resistance, should it arise. Here we develop mathematical and computational models to identify conditions under which suppression drives will evade resistance, even if resistance is present initially. Previous models assumed resistance is allelic to the drive. We relax this assumption and show that linkage between the resistance and drive loci is critical to the evolution of resistance and that evolution of resistance requires (negative) linkage disequilibrium between the two loci. When the two loci are unlinked or only partially so, a suppression drive that causes limited inviability can evolve to fixation while causing only a minor increase in resistance frequency. Once fixed, the drive allele no longer selects resistance. Our analyses suggest that among gene drives that cause moderate suppression, toxin-antidote systems are less apt to select for resistance than homing drives. Single drives of moderate effect might cause only moderate population suppression, but multiple drives (perhaps delivered sequentially) would allow arbitrary levels of suppression. The most favorable case for evolution of resistance appears to be with suppression homing drives in which resistance is dominant and fully suppresses transmission distortion; partial suppression by resistance heterozygotes or recessive resistance are less prone to resistance evolution. Given that it is now possible to engineer CRISPR-based gene drives capable of circumventing allelic resistance, this design may allow for the engineering of suppression gene drives that are effectively resistance-proof.
Structure of SARS-CoV-2 ORF8, a rapidly evolving coronavirus protein implicated in immune evasion
Authors: Flower Thomas G. , Buffalo Cosmo Z. , Hooy Richard M. , Allaire Marc , Ren Xuefeng , Hurley James H.
Published: August 2020

The molecular basis for the severity and rapid spread of the COVID-19 disease caused by SARS-CoV-2 is largely unknown. ORF8 is a rapidly evolving accessory protein that has been proposed to interfere with immune responses. The crystal structure of SARS-CoV-2 ORF8 was determined at 2.04 Å resolution by x-ray crystallography. The structure reveals a ~60 residue core similar to SARS-CoV ORF7a with the addition of two dimerization interfaces unique to SARS-CoV-2 ORF8. A covalent disulfide-linked dimer is formed through an N-terminal sequence specific to SARS-CoV-2, while a separate non-covalent interface is formed by another SARS-CoV-2-specific sequence,73YIDI76. Together the presence of these interfaces shows how SARS-CoV-2 ORF8 can form unique large-scale assemblies not possible for SARS-CoV, potentially mediating unique immune suppression and evasion activities.