bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ephyrae and metaephyrae of Pelagia noctiluca: stage determination, morphometry and shrinkage
Authors: Ottmann Daniel , Leyva Laura , Reglero Patricia , Prieto Laura , Alvarez Itziar 
Published: August 2021

Abstract
Understanding processes occurring in the different life stages of jellyfish is key to advance knowledge on their trophic interactions and population dynamics. We describe four developmental stages of Pelagia noctiluca ephyrae and metaephyrae based on the progress of feeding structures and morphometric measurements on the central disc diameter and total body diameter. Size differs significantly among stages, but it can overlap substantially, suggesting that it is not always coupled with development progress due to different somatic growth. Morphological distinction of stages is biologically important because it implies different levels of food specialization and capture efficiency. We further report a 25% (±13 SD) shrinkage of ephyrae and metaephyrae after storage in 4% formaldehyde solution. This metric can be used in ecological studies focusing on size-related traits of field observed individuals.
Multiparameter in vivo imaging in plants using genetically encoded fluorescent indicator multiplexing
Authors: Waadt Rainer , Kudla Jörg , Kollist Hannes 
Published: August 2021

Abstract
Biological processes are highly dynamic, and during plant growth, development, and environmental interactions, they occur and influence each other on diverse spatiotemporal scales. Understanding plant physiology on an organismic scale requires analyzing biological processes from various perspectives, down to the cellular and molecular levels. Ideally, such analyses should be conducted on intact and living plant tissues. Fluorescent protein (FP)-based in vivo biosensing using genetically encoded fluorescent indicators (GEFIs) is a state-of-the-art methodology for directly monitoring cellular ion, redox, sugar, hormone, ATP and phosphatidic acid dynamics, and protein kinase activities in plants. The steadily growing number of diverse but technically compatible genetically encoded biosensors, the development of dual-reporting indicators, and recent achievements in plate-reader-based analyses now allow for GEFI multiplexing: the simultaneous recording of multiple GEFIs in a single experiment. This in turn enables in vivo multiparameter analyses: the simultaneous recording of various biological processes in living organisms. Here, we provide an update on currently established direct FP-based biosensors in plants, discuss their functional principles, and highlight important biological findings accomplished by employing various approaches of GEFI-based multiplexing. We also discuss challenges and provide advice for FP-based biosensor analyses in plants.
Cerebello-Thalamo-Cortical Hyperconnectivity Classifies Patients and Predicts Long-Term Treatment Outcome in First-Episode Schizophrenia
Authors: Cao Hengyi , Wei Xia , Hu Na , Zhang Wenjing , Xiao Yuan , Zeng Jiaxin , Sweeney John A , Lencer Rebekka , Lui Su , Gong Qiyong 
Published: August 2021

Abstract
It has previously been shown that cerebello-thalamo-cortical (CTC) hyperconnectivity is likely a state-independent neural signature for psychosis. However, the potential clinical utility of this change has not yet been evaluated. Here, using fMRI and clinical data acquired from 214 untreated first-episode patients with schizophrenia (62 of whom were clinically followed-up at least once at the 12th and 24th months after treatment initiation) and 179 healthy controls, we investigated whether CTC hyperconnectivity would serve as an individualized biomarker for diagnostic classification and prediction of long-term treatment outcome. Cross-validated LASSO regression was conducted to estimate the accuracy of baseline CTC connectivity for patient-control classification, with the generalizability of classification performance tested in an independent sample including 42 untreated first-episode patients and 65 controls. Associations between baseline CTC connectivity and clinical outcomes were evaluated using linear mixed model and leave-one-out cross validation. We found significantly increased baseline CTC connectivity in patients (P = .01), which remained stable after treatment. Measures of CTC connectivity discriminated patients from controls with moderate classification accuracy (AUC = 0.68, P &lt; .001), and the classification model had good generalizability in the independent sample (AUC = 0.70, P &lt; .001). Higher CTC connectivity at baseline significantly predicted poorer long-term symptom reduction in negative symptoms (R = 0.31, P = .01) but not positive or general symptoms. These findings provide initial evidence for the putative “CTC hyperconnectivity” anomaly as an individualized diagnostic and prognostic biomarker for schizophrenia, and highlight the potential of this measure in precision psychiatry.
Alterations in Sucrose and Phenylpropanoid Metabolism Affected by BABA-Primed Defense in Postharvest Grapes and the Associated Transcriptional Mechanism
Authors: Li Chunhong , Wang Kaituo , Lei Changyi , Cao Shifeng , Huang Yixiao , Ji Nana , Xu Feng , Zheng Yonghua 
Published: August 2021

Defense elicitors can induce fruit disease resistance to control postharvest decay but may incur quality impairment. Our present work aimed to investigate the resistance against Botrytis cinerea induced by the elicitor β-aminobutyric acid (BABA) and to elucidate the specific transcriptional mechanism implicated in defense-related metabolic regulations. The functional dissection results demonstrated that, after inoculation with the fungal necrotroph B. cinerea, a suite of critical genes encoding enzymes related to the sucrose metabolism and phenylpropanoid pathway in priming defense in grapes were transcriptionally induced by treatment with 10 mM BABA. In contrast, more UDP-glucose, a shared precursor of phenylpropanoid and sucrose metabolism, may be redirected to the phenylpropanoid pathway for the synthesis of phytoalexins, including trans-resveratrol and ɛ-viniferin, in 100 mM BABA–treated grapes, resulting in direct resistance but compromised soluble sugar contents. An R2R3-type MYB protein from Vitis vinifera, VvMYB44, was isolated and characterized. VvMYB44 expression was significantly induced upon the grapes expressed defensive reaction. Subcellular localization, yeast two-hybrid, and coimmunoprecipitation assays revealed that the nuclear-localized VvMYB44 physically interacted with the salicylic acid–responsive transcription coactivator NPR1 in vivo for defense expression. In addition, VvMYB44 directly bound to the promoter regions of sucrose and phenylpropanoid metabolism-related genes and transactivated their expression, thus tipping the balance of antifungal compound accumulation and soluble sugar maintenance. Hence, these results suggest that 2R-type VvMYB44 might be a potential positive participant in BABA-induced priming defense in grape berries that contributes to avoiding the excessive consumption of soluble sugars during the postharvest storage. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .
Circadian rhythm disruption with high‐fat diet impairs glycemic control and bone quality
Authors: LLabre Joan E. , Trujillo Ruben , Sroga Grażyna E. , Figueiro Mariana G. , Vashishth Deepak 
Published: August 2021

AbstractBiological functions, including glycemic control and bone metabolism, are highly influenced by the body's internal clock. Circadian rhythms are biological rhythms that run with a period close to 24 hours and receive input from environmental stimuli, such as the light/dark cycle. We investigated the effects of circadian rhythm disruption (CRD), through alteration of the light/dark schedule, on glycemic control and bone quality of mice. Ten‐week‐old male mice (C57/BL6, n = 48) were given a low‐fat diet (LFD) or a high‐fat diet (HFD) and kept on a dayshift or altered schedule (RSS3) for 22 weeks. Mice were divided into four experimental groups (n = 12/group): Dayshift/LFD, Dayshift/HFD, RSS3/LFD, and RSS3/HFD. CRD in growing mice fed a HFD resulted in a diabetic state, with a 36.2% increase in fasting glucose levels compared to the Dayshift/LFD group. Micro‐CT scans of femora revealed a reduction in inner and outer surface expansion for mice on a HFD and altered light schedule. Cancellous bone demonstrated deterioration of bone quality as trabecular number and thickness decreased while trabecular separation increased. While HFD increased cortical bone mineral density, its combination with CRD reduced this phenomenon. The growth of mineral crystals, determined by small angle X‐ray scattering, showed HFD led to smaller crystals. Considering modifications of the organic matrix, regardless of diet, CRD exacerbated the accumulation of fluorescent advanced glycation end‐products (fAGEs) in collagen. Strength testing of tibiae showed that CRD mitigated the higher strength in the HFD group and increased brittleness indicated by lower post‐yield deflection and work‐to‐fracture. Consistent with accumulation of fAGEs, various measures of toughness were lowered with CRD, but combination of CRD with HFD protected against this decrease. Differences between strength and toughness results represent different contributions of structural and material properties of bone to energy dissipation. Collectively, these results demonstrate that combination of CRD with HFD impairs glycemic control and have complex effects on bone quality.
8‐acetoxy‐trisulfopyrene as the first activatable fluorogenic probe for add‐and‐read assessment of Organic anion‐transporting polypeptides, OATP1B1, OATP1B3, and OATP2B1
Authors: Ungvári Orsolya , Király Laura , Bakos Éva , Özvegy‐Laczka Csilla 
Published: August 2021

AbstractOrganic anion‐transporting polypeptides, OATP1B1, OATP1B3, and OATP2B1 are multispecific membrane proteins mediating the hepatocellular uptake of structurally diverse endo‐ and exogenous compounds, including various kinds of drugs. Co‐administration of OATP1B/2B1 substrates may lead to altered pharmacokinetics or even toxicity. Therefore, the study of the interaction with these OATPs is essential in drug development and is recommended by international regulatory agencies, the FDA, EMA, and PMDA. In general, radiolabeled indicators are used to measure drug interactions of OATPs, and, lately, fluorescent probes are also gaining wider application in OATP tests. However, all of the currently available methods (either radioactive or fluorescence‐based) comprise multiple steps, including the removal of the indicator in the end of the experiment. Hence, they are not ideally suited for high‐throughput screening. In the current study, in order to find an indicator allowing real‐time assessment of hepatic OATP function, we searched for an activatable fluorogenic OATP substrate. Here, we show that 8‐acetoxypyrene‐1,3,6‐trisulfonate (Ace), a fluorogenic derivative of the hepatic OATP substrate pyranine (8‐hydroxypyrene‐1,3,6‐trisulfonate) enters the cells via OATP1B1/3 or OATP2B1 function. In living cells, Ace is then converted into highly fluorescent pyranine, allowing “no‐wash” measurement of OATP function and drug interactions. Furthermore, we demonstrate that Ace can be used in an indirect assay termed as competitive counterflow suitable to distinguish between transported substrates and inhibitors of OATP1B1. The fluorescence‐based methods described here are unique and open the way toward high‐throughput screening of interactions between new molecular entities and OATPs.
A mutation in desmin makes skeletal muscle less vulnerable to acute muscle damage after eccentric loading in rats
Authors: Langer Henning T. , Mossakowski Agata A. , Avey Alec M. , Wohlgemuth Ross P. , Smith Lucas R. , Zbinden‐Foncea Herman , Baar Keith 
Published: August 2021

AbstractDesminopathy is the most common intermediate filament disease in humans. The most frequent mutation causing desminopathy in patients is a R350P DES missense mutation. We have developed a rat model with an analogous mutation in R349P Des. To investigate the role of R349P Des in mechanical loading, we stimulated the sciatic nerve of wild‐type littermates (WT) (n = 6) and animals carrying the mutation (MUT) (n = 6) causing a lengthening contraction of the dorsi flexor muscles. MUT animals showed signs of ongoing regeneration at baseline as indicated by a higher number of central nuclei (genotype: P < .0001). While stimulation did not impact central nuclei, we found an increased number of IgG positive fibers (membrane damage indicator) after eccentric contractions with both genotypes (stimulation: P < .01). Interestingly, WT animals displayed a more pronounced increase in IgG positive fibers with stimulation compared to MUT (interaction: P < .05). In addition to altered histology, molecular signaling on the protein level differed between WT and MUT. The membrane repair protein dysferlin decreased with eccentric loading in WT but increased in MUT (interaction: P < .05). The autophagic substrate p62 was increased in both genotypes with loading (stimulation: P < .05) but tended to be more elevated in WT (interaction: P = .05). Caspase 3 levels, a central regulator of apoptotic cell death, was increased with stimulation in both genotypes (stimulation: P < .01) but more so in WT animals (interaction: P < .0001). Overall, our data indicate that R349P Des rats have a lower susceptibility to structural muscle damage of the cytoskeleton and sarcolemma with acute eccentric loading.
Results of Early Proximal Femoral Osteotomy at Skeletal Maturity in Legg-Calvé-Perthes Disease: Implication for the Bypass of Fragmentation Stage
Authors: Kim Sungmin , Oh Ho-Seok , Lim Jun-Hyuk , Cho Seunghyeon , Jung Sung Taek 
Published: August 2021

Background:
The aim was to assess the long-term radiographic outcomes of patients with Legg-Calvé-Perthes disease following an early proximal femoral osteotomy (PFO) performed in avascular necrosis stage or early fragmentation stage.

Methods:
In this retrospective study, we analyzed data of 65 patients aged above 6 years at the time of diagnosis with unilateral Legg-Calvé-Perthes disease, following early PFO performed at our institution between 1979 and 2013. We observed the presence of bypassing fragmentation stage, which was classified into complete and incomplete. We compared radiographic outcomes between patients with bypass of fragmentation stage (26 hips) and those without (31 hips). Regarding skeletal maturity, the hips were graded according to the femoral head shapes: spherical, ovoid, or flat.

Results:
The mean age at diagnosis was 7.9 years (range: 6.0 to 11.9 y). The average follow-up period was 11.8 years (range: 5.9 to 22.9 y). Fragmentation bypass occurred in 40.5% (26/65) of patients, with 8 (12.3%) “undetermined” cases. Patients who completely or partially bypassed fragmentation experienced significantly less severe lateral pillar collapse (P=0.016). The femoral head was also significantly more spherical in patients with bypass (P=0.024).

Conclusions:
Our results show that 40.5% of patients who underwent early PFO bypassed the fragmentation stage. The degree of lateral pillar collapse was lower in patients with bypass. In addition, there were significantly more patients with more spherical femoral head in the bypass group.

Level of Evidence:
Level IV—case series.
When Is a Growth-friendly Strategy Warranted? A Matched Comparison of Growing Rods Versus Primary Posterior Spinal Fusion in Juveniles With Early-onset Scoliosis
Authors: Keil Lukas G. , Nash Alysa B. , Stürmer Til , Golightly Yvonne M. , Lin Feng-Chang , Stone Joseph D. , Sanders James O. , Louer Craig R. 
Published: August 2021

Background:
In 7 to 11-year-old juveniles with severe early-onset scoliosis (EOS) the optimal surgical option remains uncertain. This study compares growing rods (GRs) followed by definitive posterior spinal fusion (PSF) versus primary PSF in this population. We hypothesized that the thoracic height afforded by GRs would be offset by increased rigidity, more complications, and more operations.

Methods:
This retrospective comparative study included EOS patients aged 7.0 to 11.9 years at index surgery treated with GR→PSF or primary PSF during 2013 to 2020. Primary outcomes were thoracic height gain (ΔT1-12H), major curve, complications, and total operations. Primary PSFs were matched with replacement 1-to-n to GR→PSFs by age at index, etiology, and major curve.

Results:
Twenty-eight GR→PSFs met criteria: 19 magnetically controlled GRs and 9 traditional GRs. Three magnetically controlled GRs were definitively explanted without PSF due to complications. The remaining 25 GR→PSFs were matched to 17 primary PSFs with 100% etiology match, mean Δ major curve 1 degree, and mean Δ age at index 0.5 years (PSFs older). Median ΔT1-12H pre-GR to post-PSF was 4.7 cm with median deformity correction of 37%. Median ΔT1-12H among primary PSFs was 1.9 cm with median deformity correction of 62%. GR→PSFs had mean 1.8 complications and 3.4 operations. Primary PSFs had mean 0.5 complications and 1.3 operations. Matched analysis showed adjusted mean differences of 2.3 cm greater ΔT1-12H among GR→PSFs than their matched primary PSFs, with 25% less overall coronal deformity correction, 1.2 additional complications, and 2.2 additional operations per patient.

Conclusions:
In juveniles aged 7 to 11 with EOS, on average GRs afford 2 cm of thoracic height over primary PSF at the cost of poorer deformity correction and additional complications and operations. Primary PSF affords an average of 2 cm of thoracic height gain; if an additional 2 cm will be impactful then GRs should be considered. However, in most juveniles the height gained may not warrant the iatrogenic stiffness, complications, and additional operations. Surgeons and families should weigh these benefits and harms when choosing a treatment plan.

Level of Evidence:
Level III—retrospective comparative study.
Entire-spine Magnetic Resonance Imaging Findings and Costs in Children With Presumed Adolescent Idiopathic Scoliosis
Authors: Johnson Mitchell A. , Gohel Shivani , Mitchell Stuart L. , Flynn John (Jack) M. , Baldwin Keith D. 
Published: August 2021

Background:
Patients with adolescent idiopathic scoliosis (AIS) are commonly monitored for curve progression with spinal radiographs; however, the utility of magnetic resonance imaging (MRI) screening is unclear. The purpose of this study was to assess the findings of screening MRI for patients with a nonsurgical curve size ordered during routine clinical care and compare them with MRI ordered for patients with large curves as part of preoperative screening.

Methods:
All consecutive patients with presumed AIS who underwent entire-spine MRI with a presumed diagnosis of idiopathic scoliosis at a single institution between 2017 and 2019 were retrospectively reviewed. Patients were stratified based on MRI indication into the following groups: preoperative evaluation, pain, neurological symptoms, abnormal radiographic curve appearance, rapidly progressive curve, and other. Neural axis abnormalities recorded included concern for tethered spinal cord, syringomyelia, and Chiari malformation. The MRI findings of preoperative patients with large curves were compared with all other patients. The number needed to diagnose (NND) a neurological finding was calculated in patients whose MRIs were ordered during routine clinical care. The amount charged for each patient undergoing entire-spine MRI was determined by review of our institution’s Financial Decision Support system.

Results:
There were 344 patients included in this study with 214 (62%) MRIs performed for preoperative evaluation. Although MRI abnormalities were found in 49% of patients, only 7.0% (24/344) demonstrated neural axis abnormalities with no difference between preoperative and other indications (P=0.37). For patients with nonsurgical curves undergoing MRI due to a complaint of back pain (n=28), there were no neural axis abnormalities, and a lower rate of disk herniation/degenerative changes detected compared with preoperative MRI (3.6% vs. 18%, P=0.06). Among the 15 patients undergoing MRI for a neurological concern, 1 had a neural axis abnormality that required surgical detethering. The NND for MRI to detect a neural axis abnormality that potentially required neurosurgical intervention in nonpreoperative patients with a neurological concern was 34.4. The average cost for MRI was $17,816 (range: $2601 to $22,411) with a total cost of $2,368,439 for nonsurgical curves.

Conclusions:
Entire-spine MRI for nonpreoperative indications including pain, abnormal radiographic curve appearance, and rapid curve progression has minimal utility for patients with AIS. For patients with neurological complaints, the NND a potentially treatment-altering finding with MRI is 34.4.

Level of Evidence:
Level II—diagnostic.
Reliability and Validity of Visual Estimation of Femoral Head Hypoperfusion on Perfusion MRI in Legg-Calve-Perthes Disease
Authors: Chong David Y. , Schrader Tim , Laine Jennifer C. , Yang Scott , Gilbert Shawn R. , Kim Harry K.W. 
Published: August 2021

Background:
Assessing the severity of Legg-Calve-Perthes disease is important for treatment decision-making and determining prognosis. Perfusion magnetic resonance imaging (MRI) scans have improved our ability to assess femoral head hypoperfusion, and the use of an image analysis software called HipVasc (Scottish Rite for Children, Dallas TX) has allowed precise quantification for research purposes. In the clinical setting, using HipVasc software is not practical, and visual estimation is used to assess hypoperfusion. Currently, the reliability of visual estimation is unknown. The purpose of this study was to determine the reliability and validity of visual estimation of hypoperfusion on perfusion MRI and compare the results to measurements obtained with the HipVasc software.

Methods:
Fourteen pediatric orthopaedic surgeons, divided into 4 groups, participated in this study. Each surgeon completed a 30-minute in-person training on HipVasc before proceeding. Stratified by category of severity, 122 MRIs were randomly assigned to 1 of the 4 groups so that each group member measured the same set of ∼30 MRIs. Each surgeon documented their visual estimation of hypoperfusion first, then used HipVasc to measure the hypoperfusion and recorded their results. Interobserver reliability was assessed at the group level by calculating the intraclass correlation coefficient.

Results:
Good correlation and reliability was found between the visual estimate and HipVasc measurements with an overall mean intraclass correlation coefficient of 0.87 and mean Pearson coefficient of 0.90. The mean interobserver reliability of visual estimation was 0.84. Observers had a tendency to overestimate hypoperfusion with visual estimation.

Conclusions:
This study demonstrates the reliability and validity of visual estimation as a clinically applicable method for determining femoral head hypoperfusion. Our study is the first to measure the reliability of visual estimation and HipVasc with a large cohort of clinicians, specifically pediatric orthopaedic surgeons with varied experience in graphical software analysis. We recommend using visual estimation as a clinically applicable method to make treatment decisions, and its accuracy is comparable to HipVasc software analysis.

Level of Evidence:
Level III—diagnostic.