Generation of a Broadly Useful Model for COVID-19 Pathogenesis, Vaccination, and Treatment.

COVID-19, caused by SARS-CoV-2, is a virulent pneumonia, with >4,000,000 confirmed cases worldwide and >290,000 deaths as of May 15, 2020. It is critical that vaccines and therapeutics be developed very rapidly. Mice, the ideal animal for assessing such interventions, are resistant to SARS-CoV-2. Here, we overcome this difficulty by exogenous delivery of human ACE2 with a replication-deficient adenovirus (Ad5-hACE2). Ad5-hACE2-sensitized mice developed pneumonia characterized by weight loss, severe pulmonary pathology, and high-titer virus replication in lungs. Type I interferon, T cells, and, most importantly, signal transducer and activator of transcription 1 (STAT1) are critical for virus clearance and disease resolution in these mice. Ad5-hACE2-transduced mice enabled rapid assessments of a vaccine candidate, of human convalescent plasma, and of two antiviral therapies (poly I:C and remdesivir). In summary, we describe a murine model of broad and immediate utility to investigate COVID-19 pathogenesis and to evaluate new therapies and vaccines.

Genomic Anatomy of Homozygous XX Females and YY Males Reveals Early Evolutionary Trajectory of Sex-determining Gene and Sex Chromosomes in Silurus Fishes.

Sex chromosomes display remarkable diversity and variability among vertebrates. Compared with research on the X/Y and Z/W chromosomes, which have long evolutionary histories in mammals and birds, studies on the sex chromosomes at early evolutionary stages are limited. Here, we precisely assembled the genomes of homozygous XX female and YY male Lanzhou catfish (Silurus lanzhouensis) derived from an artificial gynogenetic family and a self-fertilized family, respectively. Chromosome 24 (Chr24) was identified as the sex chromosome based on resequencing data. Comparative analysis of the X and Y chromosomes showed an approximate 320 kb Y-specific region with a Y-specific duplicate of anti-Mullerian hormone type II receptor (amhr2y), which is consistent with findings in 2 other Silurus species but on different chromosomes (Chr24 of Silurus meridionalis and Chr5 of Silurus asotus). Deficiency of amhr2y resulted in male-to-female sex reversal, indicating that amhr2y plays a male-determining role in S. lanzhouensis. Phylogenetic analysis and comparative genomics revealed that the common sex-determining gene amhr2y was initially translocated to Chr24 of the Silurus ancestor along with the expansion of transposable elements. Chr24 was maintained as the sex chromosome in S. meridionalis and S. lanzhouensis, whereas a sex-determining region transition triggered sex chromosome turnover from Chr24 to Chr5 in S. asotus. Additionally, gene duplication, translocation, and degeneration were observed in the Y-specific regions of Silurus species. These findings present a clear case for the early evolutionary trajectory of sex chromosomes, including sex-determining gene origin, repeat sequence expansion, gene gathering and degeneration in sex-determining region, and sex chromosome turnover.

Deficiency of FRMD5 results in neurodevelopmental dysfunction and autistic-like behavior in mice.

The pathophysiology of autism spectrum disorders (ASDs) is causally linked to postsynaptic scaffolding proteins, as evidenced by numerous large-scale genomic studies [1, 2] and in vitro and in vivo neurobiological studies of mutations in animal models [3, 4]. However, due to the distinct phenotypic and genetic heterogeneity observed in ASD patients, individual mutation genes account for only a small proportion (<2%) of cases [1, 5]. Recently, a human genetic study revealed a correlation between de novo variants in FERM domain-containing-5 (FRMD5) and neurodevelopmental abnormalities [6]. In this study, we demonstrate that deficiency of the scaffolding protein FRMD5 leads to neurodevelopmental dysfunction and ASD-like behavior in mice. FRMD5 deficiency results in morphological abnormalities in neurons and synaptic dysfunction in mice. Frmd5-deficient mice display learning and memory dysfunction, impaired social function, and increased repetitive stereotyped behavior. Mechanistically, tandem mass tag (TMT)-labeled quantitative proteomics revealed that FRMD5 deletion affects the distribution of synaptic proteins involved in the pathological process of ASD. Collectively, our findings delineate the critical role of FRMD5 in neurodevelopment and ASD pathophysiology, suggesting potential therapeutic implications for the treatment of ASD.

Two duplicated gsdf homeologs cooperatively regulate male differentiation by inhibiting cyp19a1a transcription in a hexaploid fish.

Although evolutionary fates and expression patterns of duplicated genes have been extensively investigated, how duplicated genes co-regulate a biological process in polyploids remains largely unknown. Here, we identified two gsdf (gonadal somatic cell-derived factor) homeologous genes (gsdf-A and gsdf-B) in hexaploid gibel carp (Carassius gibelio), wherein each homeolog contained three highly conserved alleles. Interestingly, gsdf-A and gsdf-B transcription were mainly activated by dmrt1-A (dsx- and mab-3-related transcription factor 1) and dmrt1-B, respectively. Loss of either gsdf-A or gsdf-B alone resulted in partial male-to-female sex reversal and loss of both caused complete sex reversal, which could be rescued by a nonsteroidal aromatase inhibitor. Compensatory expression of gsdf-A and gsdf-B was observed in gsdf-B and gsdf-A mutants, respectively. Subsequently, we determined that in tissue culture cells, Gsdf-A and Gsdf-B both interacted with Ncoa5 (nuclear receptor coactivator 5) and blocked Ncoa5 interaction with Rora (retinoic acid-related orphan receptor-alpha) to repress Rora/Ncoa5-induced activation of cyp19a1a (cytochrome P450, family 19, subfamily A, polypeptide 1a). These findings illustrate that Gsdf-A and Gsdf-B can regulate male differentiation by inhibiting cyp19a1a transcription in hexaploid gibel carp and also reveal that Gsdf-A and Gsdf-B can interact with Ncoa5 to suppress cyp19a1a transcription in vitro. This study provides a typical case of cooperative mechanism of duplicated genes in polyploids and also sheds light on the conserved evolution of sex differentiation.

MSKI-RADS: An MRI-based Musculoskeletal Infection Reporting and Data System for the Diagnosis of Extremity Infections.

Background Current terms used to describe the MRI findings for musculoskeletal infections are nonspecific and inconsistent. Purpose To develop and validate an MRI-based musculoskeletal infection classification and scoring system. Materials and Methods In this retrospective cross-sectional internal validation study, a Musculoskeletal Infection Reporting and Data System (MSKI-RADS) was designed. Adult patients with radiographs and MRI scans of suspected extremity infections with a known reference standard obtained between June 2015 and May 2019 were included. The scoring categories were as follows: 0, incomplete imaging; I, negative for infection; II, superficial soft-tissue infection; III, deeper soft-tissue infection; IV, possible osteomyelitis (OM); V, highly suggestive of OM and/or septic arthritis; VI, known OM; and NOS (not otherwise specified), nonspecific bone lesions. Interreader agreement for 20 radiologists from 13 institutions (intraclass correlation coefficient [ICC]) and true-positive rates of MSKI-RADS were calculated and the accuracy of final diagnoses rendered by the readers was compared using generalized estimating equations for clustered data. Results Among paired radiographs and MRI scans from 208 patients (133 male, 75 female; mean age, 55 years ± 13 [SD]), 20 were category I; 34, II; 35, III; 30, IV; 35, V; 18, VI; and 36, NOS. Moderate interreader agreement was observed among the 20 readers (ICC, 0.70; 95% CI: 0.66, 0.75). There was no evidence of correlation between reader experience and overall accuracy (P = .94). The highest true-positive rate was for MSKI-RADS I and NOS at 88.7% (95% CI: 84.6, 91.7). The true-positive rate was 73% (95% CI: 63, 80) for MSKI-RADS V. Overall reader accuracy using MSKI-RADS across all patients was 65% ± 5, higher than final reader diagnoses at 55% ± 7 (P < .001). Conclusion MSKI-RADS is a valid system for standardized terminology and recommended management of imaging findings of peripheral extremity infections across various musculoskeletal-fellowship-trained reader experience levels. © RSNA, 2024 Supplemental material is available for this article. See also the editorial by Schweitzer in this issue.

Zebrafish TMEM47 is an effective blocker of IFN production during RNA and DNA virus infection.

IMPORTANCE: Mitochondrial antiviral signaling protein (MAVS) and stimulator of interferon (IFN) genes (STING) are key adaptor proteins required for innate immune responses to RNA and DNA virus infection. Here, we show that zebrafish transmembrane protein 47 (TMEM47) plays a critical role in regulating MAVS- and STING-triggered IFN production in a negative feedback manner. TMEM47 interacted with MAVS and STING for autophagic degradation, and ATG5 was essential for this process. These findings suggest the inhibitory function of TMEM47 on MAVS- and STING-mediated signaling responses during RNA and DNA virus infection.

Zebrafish MAP2K7 Simultaneously Enhances Host IRF7 Stability and Degrades Spring Viremia of Carp Virus P Protein via Ubiquitination Pathway.

During viral infection, host defensive proteins either enhance the host immune response or antagonize viral components directly. In this study, we report on the following two mechanisms employed by zebrafish mitogen-activated protein kinase kinase 7 (MAP2K7) to protect the host during spring viremia of carp virus (SVCV) infection: stabilization of host IRF7 and degradation of SVCV P protein. In vivo, map2k7+/- (map2k7-/- is a lethal mutation) zebrafish showed a higher lethality, more pronounced tissue damage, and more viral proteins in major immune organs than the controls. At the cellular level, overexpression of map2k7 significantly enhanced host cell antiviral capacity, and viral replication and proliferation were significantly suppressed. Additionally, MAP2K7 interacted with the C terminus of IRF7 and stabilized IRF7 by increasing K63-linked polyubiquitination. On the other hand, during MAP2K7 overexpression, SVCV P proteins were significantly decreased. Further analysis demonstrated that SVCV P protein was degraded by the ubiquitin-proteasome pathway, as the attenuation of K63-linked polyubiquitination was mediated by MAP2K7. Furthermore, the deubiquitinase USP7 was indispensable in P protein degradation. These results confirm the dual functions of MAP2K7 during viral infection. IMPORTANCE Normally, during viral infection, host antiviral factors individually modulate the host immune response or antagonize viral components to defense infection. In the present study, we report that zebrafish MAP2K7 plays a crucial positive role in the host antiviral process. According to the weaker antiviral capacity of map2k7+/- zebrafish than that of the control, we find that MAP2K7 reduces host lethality through two pathways, as follows: enhancing K63-linked polyubiquitination to promote host IRF7 stability and attenuating K63-mediated polyubiquitination to degrade the SVCV P protein. These two mechanisms of MAP2K7 reveal a special antiviral response in lower vertebrates.

Fish female-biased gene cyp19a1a leads to female antiviral response attenuation between sexes by autophagic degradation of MITA.

From insects to mammals, both innate and adaptive immune response are usually higher in females than in males, with the sex chromosome and hormonal differences considered the main reasons. Here, we report that zebrafish cyp19a1a (cytochrome P450, family 19, subfamily A, polypeptide 1a), an autosomal gene with female-biased expression, causes female fish to exhibit a lower antiviral response. First, we successfully constructed an infection model by intraperitoneal injection of spring viremia of carp virus (SVCV) into zebrafish (Danio rerio) and Carassius auratus herpesvirus (CaHV) in gibel carp (Carassius gibelio). Specifically, female fish were more vulnerable to viral infection than males, accompanied by a significantly weaker interferon (IFN) expression. After screening several candidates, cyp19a1a, which was highly expressed in female fish tissues, was selected for further analysis. The IFN expression and antiviral response were significantly higher in cyp19a1a-/- than in cyp19a1a+/+. Further investigation of the molecular mechanism revealed that Cyp19a1a targets mediator of IRF3 activation (MITA) for autophagic degradation. Interestingly, in the absence of MITA, Cyp19a1a alone could not elicit an autophagic response. Furthermore, the autophagy factor ATG14 (autophagy-related 14) was found interacted with Cyp19a1a to either promote or attenuate Cyp19a1a-mediated MITA degradation by either being overexpressed or knocked down, respectively. At the cellular level, both the normal and MITA-enhanced cellular antiviral responses were diminished by Cyp19a1a. These findings demonstrated a sex difference in the antiviral response based on a regulation mechanism controlled by a female-biased gene besides sex chromosome and hormonal differences, supplying the current understanding of sex differences in fish.

CT-based comprehensive scoring system for assessing hip arthropathy in ankylosing spondylitis: feasibility study with clinical correlation.

OBJECTIVE: To develop a CT-based scoring system for assessment of hip arthropathy in AS. METHODS: All AS patients were prospectively recruited, consented, and underwent whole-body stereoradiographs and pelvis CT, which were assessed by two independent radiologists. Stereoradiographs were assessed according to Kellgreen-Lawrence and BASRI-h. For the Hip arthropathy CT score in AS (HACTSAS), joints were divided into 7 segments and scored for joint space, osteophytes, subchondral cysts/erosions. Patients were clinically assessed for range of motion (ROM), pain, and clinical scores (BASMI, BASFI, ASQol, BASDAI and ASDAS). Radiological scores correlations with clinical parameters were compared. ROM sensitivity and specificity for hip arthropathy (BASRI-h ≥ 2) were calculated. RESULTS: Sample included 112 patients, with 36/112 females and 76/112 males. Average age was 51.0 ± 11.2 years and mean duration of AS was 20.9 ± 9.6 years. ICC for HACTSAS, Kellgreen-Lawrence and BASRI-h were 0.89, 0.89 and 0.82 respectively. HACTSAS showed moderate absolute correlation with ROM (ρ=-0.41) and BASMI (ρ = 0.45), and weak with pain (ρ = 0.18) and BASFI (ρ = 0.25). BASRI-h and Kellgreen-Lawrence exhibited moderate correlation with ROM (ρ=-0.44 and ρ=-0.40, respectively), weak with pain (ρ=-.27and ρ=-0.23, respectively) and BASFI (ρ=-0.16 and ρ=-0.18, respectively), but only weak with BASMI (ρ=-0.34 and ρ=-0.36, respectively). Internal rotation <15°, abduction <31°, and intermalleolar distance <75cm were, respectively, 73%, 70% and 73% sensitivity and 81%, 65% and 68% specific for hip arthropathy. CONCLUSION: HACTSAS exhibited higher correlation with BASMI and BASFI when compared with BASRI-h, but less correlation with pain and ROM. Internal rotation was the best clinical discriminator for hip arthropathy.

3D color-rendered MR neurography heatmaps in visualizing normal lumbosacral (LS) plexus and increasing conspicuity of LS plexopathy.

PURPOSE: To determine whether color-rendered 3D MR neurography (MRN) images (heatmaps) improve diagnostic accuracy, reader confidence levels, and time savings to assess LS plexus lesions compared to the conventional grayscale images. MATERIALS AND METHODS: A cross-sectional study included adults of all genders with randomly chosen MRNs of LS plexus and known reference standards of normal or neuropathy (plexopathy and radiculopathy). Heatmaps were constructed using 3D MRN STIR images and color rendered with higher intensity to yellow and lower intensity to darker-red colors in 1-2 min on average and were available on PACS for the readers. 2D plus 3D grayscale MIP images and 2D plus 3D MIP heatmaps were analyzed by four musculoskeletal radiologists (two faculty and two fellows) in two separate rounds blinded to the final diagnosis. Readers evaluated: neuropathy and number of nerves affected (neuropathy score: 0-normal; 1-one nerve affected; and 2-two or more nerves affected); final diagnosis; confidence levels; and time taken to evaluate the studies. Conger's kappa and paired t-test were used for analysis. RESULTS: Among 70 MRNs from 70 patients, there were 32 males and 38 females with average age ± SD of 54.8 ± 20.1 years and 49.9 ± 16.6 years, respectively. There were 30 normal and 40 LS plexus lesion scans. Interreader agreements for neuropathy scores were substantial to moderate on conventional imaging and heat maps (Conger's kappa: 0.65; 95% CI: 0.55, 0.73, and 0.59; 95% CI: 0.47, 0.69), respectively. The mean neuropathy score and final diagnosis accuracies were similar in both rounds 85.7% ± 0.1% vs 83.2% ± 0.1% (p = 0.13), and 83.6% ± 0.1% vs 80.0% ± 0.1%; p = 0.16), respectively. Time savings were significant when using heatmaps for all readers (p < 0.001). Time savings using heatmaps ranged from 57.7% to 74.6% and 56.3% to 75% of the original time for the fellows and faculty, respectively. Average confidence levels for neuropathy score significantly increased using heatmaps for one fellow and one faculty (p < 0.05), while average confidence levels for final diagnosis improved for both fellows and one faculty (p < 0.05). CONCLUSION: 3D color-rendered MRN heatmaps show comparable diagnostic accuracy to conventional MRN imaging but with significant time savings to identify LS plexus lesions. KEY POINTS: Question Do color-rendered 3D MRN images (heatmaps) improve accuracy, and confidence, and save time when assessing lumbosacral (LS) plexus lesions compared to conventional grayscale images? Findings 3D-rendered heatmaps showed comparable diagnostic accuracy with time savings ranging from 56.3% to 75%. Clinical relevance 3D color-rendered heatmaps increase time efficiency in evaluating MRNs of LS plexus, allowing for improved radiologist productivity and diagnostic confidence.

MRI-based Neuropathy Score Reporting And Data System (NS-RADS): multi-institutional wider-experience usability study of peripheral neuropathy conditions among 32 radiology readers.

OBJECTIVE: To determine the inter-reader reliability and diagnostic performance of classification and severity scales of Neuropathy Score Reporting And Data System (NS-RADS) among readers of differing experience levels after limited teaching of the scoring system. METHODS: This is a multi-institutional, cross-sectional, retrospective study of MRI cases of proven peripheral neuropathy (PN) conditions. Thirty-two radiology readers with varying experience levels were recruited from different institutions. Each reader attended and received a structured presentation that described the NS-RADS classification system containing examples and reviewed published articles on this subject. The readers were then asked to perform NS-RADS scoring with recording of category, subcategory, and most likely diagnosis. Inter-reader agreements were evaluated by Conger's kappa and diagnostic accuracy was calculated for each reader as percent correct diagnosis. A linear mixed model was used to estimate and compare accuracy between trainees and attendings. RESULTS: Across all readers, agreement was good for NS-RADS category and moderate for subcategory. Inter-reader agreement of trainees was comparable to attendings (0.65 vs 0.65). Reader accuracy for attendings was 75% (95% CI 73%, 77%), slightly higher than for trainees (71% (69%, 72%), p = 0.0006) for nerves and comparable for muscles (attendings, 87.5% (95% CI 86.1-88.8%) and trainees, 86.6% (95% CI 85.2-87.9%), p = 0.4). NS-RADS accuracy was also higher than average accuracy for the most plausible diagnosis for attending radiologists at 67% (95% CI 63%, 71%) and for trainees at 65% (95% CI 60%, 69%) (p = 0.036). CONCLUSION: Non-expert radiologists interpreted PN conditions with good accuracy and moderate-to-good inter-reader reliability using the NS-RADS scoring system. CLINICAL RELEVANCE STATEMENT: The Neuropathy Score Reporting And Data System (NS-RADS) is an accurate and reliable MRI-based image scoring system for practical use for the diagnosis and grading of severity of peripheral neuromuscular disorders by both experienced and general radiologists. KEY POINTS: • The Neuropathy Score Reporting And Data System (NS-RADS) can be used effectively by non-expert radiologists to categorize peripheral neuropathy. • Across 32 different experience-level readers, the agreement was good for NS-RADS category and moderate for NS-RADS subcategory. • NS-RADS accuracy was higher than the average accuracy for the most plausible diagnosis for both attending radiologists and trainees (at 75%, 71% and 65%, 65%, respectively).

Cost-Effectiveness of an Ultrasound-First Strategy in the Diagnostic Evaluation of Noncalcified Lesions Recalled From Screening Digital Breast Tomosynthesis.

BACKGROUND. Ultrasound may be sufficient in the diagnostic evaluation of many noncalcified lesions recalled from screening digital breast tomosynthesis (DBT). In some scenarios, omission of diagnostic mammography can save health care costs. OBJECTIVE. The purpose of this study was to evaluate the cost-effectiveness of a strategy of performing ultrasound first versus diagnostic mammography first in the diagnostic evaluation of noncalcified lesions recalled from screening DBT. METHODS. Decision tree analysis was performed to compare ultrasound first versus diagnostic mammography first in the diagnostic evaluation of DBT-recalled noncalcified lesions from a U.S. health care system perspective with a 40-year horizon. The analysis used probabilities and prevalence information from published single-institution prospective data, additional literature-derived estimates of diagnostic test performance, and Medicare-allowable reimbursement rates. Health states were represented in a Markov chain model. For each strategy, the total cost and effectiveness (expressed in quality-adjusted life-years [QALYs]) were estimated. Cost-effectiveness was assessed through incremental cost-effectiveness ratios (ICERs) and incremental net monetary benefit, with use of a willingness-to-pay (WTP) threshold of US$100,000 per QALY gained. Deterministic sensitivity analyses were performed to estimate the impact of different input parameters, and probabilistic sensitivity analysis with Monte Carlo simulations was conducted to estimate the impact of combined uncertainty across parameters. RESULTS. In the base-case scenario, for diagnostic evaluation of DBT-recalled noncalcified lesions, a strategy of performing ultrasound first versus diagnostic mammography first resulted in more cost savings (total cost, US$17,672 vs US$18,323) and greater cost-effectiveness (QALYs, 23.1309 vs 23.1306) over the 40-year horizon. The ultrasound-first strategy resulted in an ICER of -2,170,000 (expressed as U.S. dollars per QALY) and an incremental net monetary benefit of US$681 versus the diagnostic mammography-first strategy. Therefore, performing ultrasound first was deemed the more cost-effective strategy at the WTP threshold. In deterministic sensitivity analyses, the most important driver of cost-effectiveness was lost utility from delayed diagnosis, followed by the relative sensitivities of ultrasound and diagnostic mammography. In probabilistic sensitivity analysis, ultrasound first was the better strategy in 93.0% of iterations. CONCLUSION. A strategy of performing ultrasound first, with or without diagnostic mammography, is more cost-effective than a traditional strategy of conducting diagnostic mammography first. CLINICAL IMPACT. This cost-effectiveness analysis supports the growing prioritization of ultrasound as the primary method for evaluating DBT-recalled noncalcified lesions.

Understanding and Mitigating Bias in Imaging Artificial Intelligence.

Artificial intelligence (AI) algorithms are prone to bias at multiple stages of model development, with potential for exacerbating health disparities. However, bias in imaging AI is a complex topic that encompasses multiple coexisting definitions. Bias may refer to unequal preference to a person or group owing to preexisting attitudes or beliefs, either intentional or unintentional. However, cognitive bias refers to systematic deviation from objective judgment due to reliance on heuristics, and statistical bias refers to differences between true and expected values, commonly manifesting as systematic error in model prediction (ie, a model with output unrepresentative of real-world conditions). Clinical decisions informed by biased models may lead to patient harm due to action on inaccurate AI results or exacerbate health inequities due to differing performance among patient populations. However, while inequitable bias can harm patients in this context, a mindful approach leveraging equitable bias can address underrepresentation of minority groups or rare diseases. Radiologists should also be aware of bias after AI deployment such as automation bias, or a tendency to agree with automated decisions despite contrary evidence. Understanding common sources of imaging AI bias and the consequences of using biased models can guide preventive measures to mitigate its impact. Accordingly, the authors focus on sources of bias at stages along the imaging machine learning life cycle, attempting to simplify potentially intimidating technical terminology for general radiologists using AI tools in practice or collaborating with data scientists and engineers for AI tool development. The authors review definitions of bias in AI, describe common sources of bias, and present recommendations to guide quality control measures to mitigate the impact of bias in imaging AI. Understanding the terms featured in this article will enable a proactive approach to identifying and mitigating bias in imaging AI. Published under a CC BY 4.0 license. Test Your Knowledge questions for this article are available in the supplemental material. See the invited commentary by Rouzrokh and Erickson in this issue.

Investigation of the mechanism of poly (ADP-ribose) polymerase (PARP) in elderly mouse myocardial ischemia-reperfusion injury.

This study investigated the role of Poly (ADP-ribose) Polymerase (PARP) in myocardial ischemia-reperfusion injury (MIRI) in elderly mice. It involves 30 elderly male KM mice divided into three groups: Sham, MIRI, and DPQ, where the MIRI and DPQ groups undergo myocardial ischemia-reperfusion with the DPQ group also receiving DPQ for PARP-1 inhibition. Over three weeks, assessments include histological analysis of myocardial lesions, left ventricular ejection fraction (LVEF) measurements, and evaluations of serum cardiac enzymes and inflammatory markers. This approach aims to understand the protective effects of DPQ in MIRI, focusing on its impact on cardiac health and inflammation via the JAK2/STAT3 pathway. The findings suggest that PARP activation exacerbates cardiac dysfunction and inflammation in MIRI by possibly modulating the JAK2/STAT3 signaling pathway. Inhibition of PARP-1 with DPQ mitigates these effects, as indicated by reduced myocardial lesions and inflammatory infiltration, improved LVEF, and altered levels of inflammatory markers and signaling molecules. However, the differences in STAT3 and p-STAT3 protein expression between the DPQ and MIRI groups were not statistically significant, suggesting that while PARP inhibition affects many aspects of MIRI pathology, its impact on the JAK2/STAT3 pathway may not fully explain the observed benefits. This study contributes to our understanding of the complex mechanisms underlying myocardial ischemia-reperfusion injury, particularly in the context of aging. It highlights the potential of PARP inhibition as a therapeutic strategy to attenuate cardiac dysfunction and inflammation in MIRI, though further research is necessary to fully elucidate the underlying molecular pathways and to explore the clinical relevance of these findings in humans.

Three-dimensional Volumetric Visceral and Subcutaneous Fat Analysis on Opportunistic Computed Tomography Imaging of Patients With Greater Trochanteric Pain Syndrome Compared With Those With Predominant Osteoarthritis: A Case-Control Study.

OBJECTIVE: This study aimed to address the gap in knowledge assessing the impact of visceral and subcutaneous body fat on 3-dimensional computed tomography imaging in patients with greater trochanteric pain syndrome (GTPS) in comparison with those primarily diagnosed with osteoarthritis (OA). MATERIALS AND METHODS: We evaluated adult patients with a confirmed diagnosis of GTPS from our institutional hip-preservation clinic spanning 2011 to 2022. Selection criteria included their initial clinic visit for hip pain and a concurrent pelvis computed tomography scan. These patients were age- and sex-matched to mild-moderate OA patients selected randomly from the database. Visceral and subcutaneous fat areas were measured volumetrically from the sacroiliac joint to the lesser trochanter using an independent software. Interreader reliability was also calculated. RESULTS: A total of 93 patients met the study criteria, of which 37 belonged to the GTPS group and 56 belonged to the OA group. Both groups were sex and race matched. Average age in GTPS and OA groups was 59.3 years and 56 years, respectively. For GTPS group, average body mass index was 28.9 kg/m 2 , and for the OA group, average body mass index was 29.9 kg/m 2 , with no significant difference ( P > 0.05). Two-sample t test showed no significant differences in the visceral fat, subcutaneous fat, or the visceral fat to total fat volume ratio between the GTPS and OA groups. There was excellent interreader reliability. CONCLUSIONS: Our results indicate that there is no significant difference in fat distribution and volumes among GTPS and OA patients. This suggests that being overweight or obese may not be directly linked or contribute to the onset of GTPS. Other factors, such as gluteal tendinopathy, bursitis, or iliotibial band syndrome, might be responsible and need further investigation.

Single-Plane 3-Dimensional Isotropic Spin-Echo Magnetic Resonance Imaging Reconstructions of Shoulder Exhibit Superior Correlation to Surgical Findings Than 2-Dimensional Dixon Multiplanar Magnetic Resonance Imaging.

OBJECTIVE: The aim of the study is to evaluate concordance of multiplanar 2-dimensional magnetic resonance imaging (2D-MRI) versus 3D isotropic MRI for rotator cuff and labral tears with the reference standard of arthroscopic surgical findings. METHODS: It was an institutional review board-approved retrospective single-center study of consecutive preoperative patients with isotropic 3D-MRI on 3-Tesla scanners, multiplanar 2D-MRI, and shoulder arthroscopy. Scapular plane-oriented contiguous multiplanar reconstructions of 3D-images were evaluated by 2 experienced fellowship-trained musculoskeletal radiologists. Variables included the following: labral tear presence and rotator-cuff tear Ellman grade, thickness, and width. Sensitivities (Sen) and specificities (Spe) were calculated for binary variables. Mean squared errors (MSE) were calculated for ordinal variables. Lower MSE indicated higher concordance. RESULTS: Seventy-two patients (43 female) with a mean age of 50.75 ± 9.76 years were evaluated. For infraspinatus-tear presence, 3D-MRI showed higher sensitivity (0.96) and specificity (0.68) than 2D-MRI (Sen = 0.85, Spe = 0.32) ( Psen = 0.005, Pspe = 0.002). For subscapularis-tear presence, 3D-MRI showed higher sensitivity (0.94) and specificity (0.73) compared with 2D-MRI (Sen = 0.83, Spe = 0.56) ( Psen = 0.02, Pspe = 0.04). For supraspinatus-tear presence, there was no significant difference between 3D-MRI (Sen =0.96, Spe = 0.67) compared with 2D-MRI (Sen = 0.98, Spe = 0.83) ( Psen = 0.43, Pspe = 0.63). For infraspinatus-tear thickness, 3D-MRI showed lower MSE (0.35) compared with 2D-MRI MSE (0.82) ( P = 0.01). For subscapularis-tear thickness, 3D-MRI had lower MSE (0.31) compared with 2D-MRI MSE (0.51) ( P = 0.007). However, no difference noted for supraspinatus-tear thickness when comparing 3D-MRI MSE (0.39) and 2D-MRI MSE (0.51) ( P = 0.49). For labral-tear presence, 3D-MRI had a lower MSE (0.20) compared with 2D-MRI MSE (0.57) ( P < 0.001). CONCLUSIONS: Three-dimensional MRI of the shoulder is time efficient with a shorter acquisition time and exhibits comparable with superior correlation to surgical findings than 2D-MRI for detection of labral tears and some rotator cuff tears. Three-dimensional MRI may be used in place of traditional 2D-MRI in detection of soft-tissue shoulder injury in centers equipped to do so.

Characterization of Demographical Histologic Diversity in Small Renal Masses With the Clear Cell Likelihood Score.

OBJECTIVE: This study aimed to develop a diagnostic model to estimate the distribution of small renal mass (SRM; ≤4 cm) histologic subtypes for patients with different demographic backgrounds and clear cell likelihood score (ccLS) designations. MATERIALS AND METHODS: A bi-institution retrospective cohort study was conducted where 347 patients (366 SRMs) underwent magnetic resonance imaging and received a ccLS before pathologic confirmation between June 2016 and November 2021. Age, sex, race, ethnicity, socioeconomic status, body mass index (BMI), and the ccLS were tabulated. The socioeconomic status for each patient was determined using the Area Deprivation Index associated with their residential address. The magnetic resonance imaging-derived ccLS assists in the characterization of SRMs by providing a likelihood of clear cell renal cell carcinoma (ccRCC). Pathological subtypes were grouped into four categories (ccRCC, papillary renal cell carcinoma, other renal cell carcinomas, or benign). Generalized estimating equations were used to estimate probabilities of the pathological subtypes across different patient subgroups. RESULTS: Race and ethnicity, BMI, and ccLS were significant predictors of histology (all P < 0.001). Obese (BMI, ≥30 kg/m 2 ) Hispanic patients with ccLS of ≥4 had the highest estimated rate of ccRCC (97.1%), and normal-weight (BMI, <25 kg/m 2 ) non-Hispanic Black patients with ccLS ≤2 had the lowest (0.2%). The highest estimated rates of papillary renal cell carcinoma were found in overweight (BMI, 25-30 kg/m 2 ) non-Hispanic Black patients with ccLS ≤2 (92.3%), and the lowest, in obese Hispanic patients with ccLS ≥4 (<0.1%). CONCLUSIONS: Patient race, ethnicity, BMI, and ccLS offer synergistic information to estimate the probabilities of SRM histologic subtypes.

Genomic anatomy of male-specific microchromosomes in a gynogenetic fish.

Unisexual taxa are commonly considered short-lived as the absence of meiotic recombination is supposed to accumulate deleterious mutations and hinder the creation of genetic diversity. However, the gynogenetic gibel carp (Carassius gibelio) with high genetic diversity and wide ecological distribution has outlived its predicted extinction time of a strict unisexual reproduction population. Unlike other unisexual vertebrates, males associated with supernumerary microchromosomes have been observed in gibel carp, which provides a unique system to explore the rationales underlying male occurrence in unisexual lineage and evolution of unisexual reproduction. Here, we identified a massively expanded satellite DNA cluster on microchromosomes of hexaploid gibel carp via comparing with the ancestral tetraploid crucian carp (Carassius auratus). Based on the satellite cluster, we developed a method for single chromosomal fluorescence microdissection and isolated three male-specific microchromosomes in a male metaphase cell. Genomic anatomy revealed that these male-specific microchromosomes contained homologous sequences of autosomes and abundant repetitive elements. Significantly, several potential male-specific genes with transcriptional activity were identified, among which four and five genes displayed male-specific and male-biased expression in gonads, respectively, during the developmental period of sex determination. Therefore, the male-specific microchromosomes resembling common features of sex chromosomes may be the main driving force for male occurrence in gynogenetic gibel carp, which sheds new light on the evolution of unisexual reproduction.

Deep learning generated lower extremity radiographic measurements are adequate for quick assessment of knee angular alignment and leg length determination.

PURPOSE: Angular and longitudinal deformities of leg alignment create excessive stresses across joints, leading to pain and impaired function. Multiple measurements are used to assess these deformities on anteroposterior (AP) full-length radiographs. An artificial intelligence (AI) software automatically locates anatomical landmarks on AP full-length radiographs and performs 13 measurements to assess knee angular alignment and leg length. The primary aim of this study was to evaluate the agreements in LLD and knee alignment measurements between an AI software and two board-certified radiologists in patients without metal implants. The secondary aim was to assess time savings achieved by AI. METHODS: The measurements assessed in the study were hip-knee-angle (HKA), anatomical-tibiofemoral angle (aTFA), anatomical-mechanical-axis angle (AMA), joint-line-convergence angle (JLCA), mechanical-lateral-proximal-femur-angle (mLPFA), mechanical-lateral-distal-femur-angle (mLDFA), mechanical-medial-proximal-tibia-angle (mMPTA), mechanical-lateral-distal-tibia- angle (mLDTA), femur length, tibia length, full leg length, leg length discrepancy (LLD), and mechanical axis deviation (MAD). These measurements were performed by two radiologists and the AI software on 164 legs. Intraclass-correlation-coefficients (ICC) and Bland-Altman analyses were used to assess the AI's performance. RESULTS: The AI software set incorrect landmarks for 11/164 legs. Excluding these cases, ICCs between the software and radiologists were excellent for 12/13 variables (11/13 with outliers included), and the AI software met performance targets for 11/13 variables (9/13 with outliers included). The mean reading time for the AI algorithm and two readers, respectively, was 38.3, 435.0, and 625.0 s. CONCLUSION: This study demonstrated that, with few exceptions, this AI-based software reliably generated measurements for most variables in the study and provided substantial time savings.

Unveiling adcyap1 as a protective factor linking pain and nerve regeneration through single-cell RNA sequencing of rat dorsal root ganglion neurons.

BACKGROUND: Severe peripheral nerve injury (PNI) often leads to significant movement disorders and intractable pain. Therefore, promoting nerve regeneration while avoiding neuropathic pain is crucial for the clinical treatment of PNI patients. However, established animal models for peripheral neuropathy fail to accurately recapitulate the clinical features of PNI. Additionally, researchers usually investigate neuropathic pain and axonal regeneration separately, leaving the intrinsic relationship between the development of neuropathic pain and nerve regeneration after PNI unclear. To explore the underlying connections between pain and regeneration after PNI and provide potential molecular targets, we performed single-cell RNA sequencing and functional verification in an established rat model, allowing simultaneous study of the neuropathic pain and axonal regeneration after PNI. RESULTS: First, a novel rat model named spared nerve crush (SNC) was created. In this model, two branches of the sciatic nerve were crushed, but the epineurium remained unsevered. This model successfully recapitulated both neuropathic pain and axonal regeneration after PNI, allowing for the study of the intrinsic link between these two crucial biological processes. Dorsal root ganglions (DRGs) from SNC and naïve rats at various time points after SNC were collected for single-cell RNA sequencing (scRNA-seq). After matching all scRNA-seq data to the 7 known DRG types, we discovered that the PEP1 and PEP3 DRG neuron subtypes increased in crushed and uncrushed DRG separately after SNC. Using experimental design scRNA-seq processing (EDSSP), we identified Adcyap1 as a potential gene contributing to both pain and nerve regeneration. Indeed, repeated intrathecal administration of PACAP38 mitigated pain and facilitated axonal regeneration, while Adcyap1 siRNA or PACAP6-38, an antagonist of PAC1R (a receptor of PACAP38) led to both mechanical hyperalgesia and delayed DRG axon regeneration in SNC rats. Moreover, these effects can be reversed by repeated intrathecal administration of PACAP38 in the acute phase but not the late phase after PNI, resulting in alleviated pain and promoted axonal regeneration. CONCLUSIONS: Our study reveals that Adcyap1 is an intrinsic protective factor linking neuropathic pain and axonal regeneration following PNI. This finding provides new potential targets and strategies for early therapeutic intervention of PNI.