Semiquantitative Scoring System Research Articles

Defining the clinical validity of genes reported to cause pulmonary arterial hypertension

PurposePulmonary arterial hypertension (PAH) is a rare, progressive vasculopathy with significant cardiopulmonary morbidity and mortality. Genetic testing is currently recommended for adults diagnosed with heritable, idiopathic, anorexigen-, hereditary hemorrhagic telangiectasia–, and congenital heart disease–associated PAH, PAH with overt features of venous/capillary involvement, and all children diagnosed with PAH. Variants in at least 27 genes have putative evidence for PAH causality. Rigorous assessment of the evidence is needed to inform genetic testing. MethodsAn international panel of experts in PAH applied a semi-quantitative scoring system developed by the NIH Clinical Genome Resource to classify the relative strength of evidence supporting PAH gene-disease relationships based on genetic and experimental evidence. ResultsTwelve genes (BMPR2, ACVRL1, ATP13A3, CAV1, EIF2AK4, ENG, GDF2, KCNK3, KDR, SMAD9, SOX17, and TBX4) were classified as having definitive evidence and 3 genes (ABCC8, GGCX, and TET2) with moderate evidence. Six genes (AQP1, BMP10, FBLN2, KLF2, KLK1, and PDGFD) were classified as having limited evidence for causal effects of variants. TOPBP1 was classified as having no known PAH relationship. Five genes (BMPR1A, BMPR1B, NOTCH3, SMAD1, and SMAD4) were disputed because of a paucity of genetic evidence over time. ConclusionWe recommend that genetic testing includes all genes with definitive evidence and that caution be taken in the interpretation of variants identified in genes with moderate or limited evidence. Genes with no known evidence for PAH or disputed genes should not be included in genetic testing.

Poster 300: A Novel Magnetic Resonance Imaging (MRI) Scoring System to Evaluate Patellofemoral Osteoarthritis

Objectives: Knee osteoarthritis (OA) is a common degenerative or post-traumatic condition, and may affect one or more compartments, including the medial and lateral tibiofemoral or patellofemoral articulations. Isolated patellofemoral osteoarthritis (PFOA) has been reported in 24% of women and 11% of men aged > 55 years presenting for evaluation of knee pain. PFOA is a result of abnormal mechanical loading across the patellofemoral joint, often related to prior instability, trauma, malalignment or post- surgical changes. Treatment of PFOA typically includes a trial of conservative management, including physical therapy, injections and activity modification. Operative treatment involves joint preservation (i.e., cartilage restoration procedures, tibial tubercle osteotomy, or lateral lengthening) and joint replacement (patellofemoral arthroplasty) procedures. In contrast to the tibiofemoral compartments, the current literature lacks a reliable magnetic resonance imaging (MRI) scoring system to quantitatively evaluate patellofemoral cartilage degeneration. The present study aims to describe, apply and assess the inter-observer reliability of a novel MRI scoring system for assessment of the patellofemoral cartilage. Methods: We conducted a query of an institutional database to identify all patients who underwent patellofemoral arthroplasty by the senior surgeon (S.M.S.) between January 2012 and December 2021. Patients with poor quality or lacking preoperative knee MRI were excluded. MRI scans were performed on 1.5 or 3.0-Tesla magnets utilizing dedicated extremity coils and standardized clinical imaging protocols consistent with International Cartilage Regeneration and Joint Preservation Society (ICRS) guidelines for the assessment of articular cartilage. Preoperative MRI scans were reviewed and patellofemoral cartilage was scored by a fellowship-trained musculoskeletal radiologist and a fellowship- trained orthopedic knee surgeon using our novel MRI scoring system. The novel semi-quantitative MRI patellofemoral scoring system includes assessment of patellar and trochlear cartilage, bony architecture, patellofemoral alignment, and synovitis (Table 1). For statistical analysis, inter-observer agreement was assessed using Cohen-Kappa coefficient for categorical variables, and intraclass correlation coefficient (ICC) for the total score. The measurement agreement for reach variable and total score was interpreted according to the Landis and Koch criteria: 0 - 0.2, slight; 0.21 - 0.4, fair; 0.41 - 0.6, moderate; 0.61 - 0.80, substantial; 0.81 - 1.0, almost perfect. Results: A total of 72 patients were included in the final analysis. The mean (± standard deviation) age and body mass index (BMI) of all patients was 53.30±7.72 years and 27.3±5.37 kg/m, respectively. Patellar bone edema and patellar cysts demonstrated moderate inter-observer agreement. All cartilage features, trochlear bone edema and cysts, osteophytes, subchondral remodeling, patellar height, and synovitis demonstrated substantial inter-observer agreement. Patellofemoral alignment and total score demonstrated almost perfect inter-observer agreement (Table 2 and Figure 1). Conclusions: The novel MRI scoring system for PFOA proposed by the authors is an attempt to develop a comprehensive method to assess patellofemoral joint cartilage degeneration, including evaluation of patellar and trochlear cartilage changes, bony architecture, patellofemoral alignment, and synovitis. Importantly, all features of the scoring system demonstrated at least moderate inter-observer agreement, with the majority of variables and total score demonstrating substantial to near perfect agreement. Future studies should aim to validate this scoring system in patients with and without patellofemoral arthritis and attempt to correlate scores with clinical outcomes. [Table: see text][Table: see text]

Intramedullary bone tissue reaction of ion-releasing resin-modified glass-ionomer restoration versus two calcium silicate-based cements: an animal study

This comparative study was conducted to assess the intramedullary bone tissue reaction of an ion-releasing resin modified glass-ionomer cement with claimed bioactivity (ACTIVA bioactive resin) restorative material versus Mineral Trioxide Aggregate High Plasticity (MTA HP) and bioceramic putty iRoot BP Plus. Fifty-six adult male Wistar rats were assigned into 4 equal groups (14 rats each). A surgical intramedullary bi-lateral tibial bone defects were performed in rats of the control group I (GI) and left without any treatment to be considered as controls (n = 28). The rats of groups II, III and IV were handled as group I except that the tibial bone defects were filled with ACTIVA, MTA HP and iRoot BP, respectively. In all groups, rats were euthanized after one month and specimens were processed to histological investigation, SEM examination and EDX elemental analysis. In addition, semi-quantitative histomorphometric scoring system was conducted for the following parameters; new bone formation, inflammatory response, angiogenesis, granulation tissue, osteoblasts and osteoclasts. The clinical follow-up outcome of this study revealed the recovery of rats after 4 days post-surgical procedure. It was observed that the animal subjects returned to their routine activities, e.g., walking, grooming and eating. The rats showed normal chewing efficiency without any weight loss or postoperative complications. Histologically, the control group sections showed scanty, very thin, new bone trabeculae of immature woven type located mostly at the peripheral part of the tibial bone defects. These defects exhibited greater amount of thick bands of typically organized granulation tissue with central and peripheral orientation. Meanwhile, bone defects of ACTIVA group showed an empty space surrounded by thick, newly formed, immature woven bone trabeculae. Moreover, bone defects of MTA HP group were partially filled with thick newly formed woven bone trabeculae with wide marrow spaces presented centrally and at the periphery with little amount of mature granulation tissue at the central part. The iRoot BP Plus group section exhibited an observable woven bone formation of normal trabecular structures with narrow marrow spaces presented centrally and at the periphery showed lesser amount of well-organized/mature granulation tissue formation. Kruskal Wallis test revealed total significant differences between the control, ACTIVA, MTAHP and iRoot BP Plus groups (p < 0.05). Meanwhile, Mann–Whitney U test showed significant difference between control and ACTIVA groups, Control and MTA HP groups, control and iRoot BP Plus groups. ACTIVA and MTA HP groups, ACTIVA and iRoot BP Plus (p ˂ 0.05) with no significant difference between MTA HP and iRoot BP Plus (p > 0.05). The elemental analysis outcome showed that the lesions of the control group specimens were filled with recently created trabecular bone with limited marrow spaces. EDX tests (Ca and P analysis) indicated a lower degree of mineralization. Lower amounts of Ca and P was expressed in the mapping analysis compared with other test groups. Calcium silicate-based cements induce more bone formation when compared to an ion-releasing resin modified glass-ionomer restoration with claimed bioactivity. Moreover, the bio-inductive properties of the three tested materials are likely the same. Clinical significance: bioactive resin composite can be used as a retrograde filling.

Circulating Biomarkers and Disease Activity in Systemic Sclerosis-Associated Interstitial Lung Disease

Background: Systemic sclerosis-associated interstitial lung disease (SSc-ILD) is a major cause of death in SSc patients. Several studies have reported that serum anti-Scl-70, Krebs von Lungren-6 (KL-6), and surfactant protein D (SP-D) levels are associated with the presence and progression of ILD in SSc patients. Objective: To examine the correlation between levels of these serum biomarkers and disease severity determined by baseline dyspnea index (BDI), high-resolution computed tomography (HRCT) score, and pulmonary function tests. Materials and Methods: The present study was a single-center, cross-sectional study. Serum anti-Scl-70, KL-6, and SP-D from 20 SSc-ILD patients and five non-ILD subjects were measured. The BDI, HRCT score, and pulmonary function tests were used to assess the severity of ILD in SSc-ILD patients. HRCT abnormalities, including ground-glass opacity (GGO), fibrosis, and honeycombing, were scored by using the semi-quantitative scoring system. Results: Serum anti-Scl-70, KL-6, and SP-D in SSc-ILD patients were significantly higher than those in non-ILD subjects. There was a moderate correlation between diffusing capacity for carbon monoxide (DLCO) and serum KL-6 levels (r=–0.551, p=0.022), while the pulmonary fibrosis (PF) score exhibited a strong correlation with serum KL-6 levels (r=0.630, p=0.003). The PF score had a moderate negative correlation with forced vital capacity (FVC) (r=–0.515, p=0.034) and a strong negative correlation with total lung capacity (TLC) and DLCO (r=–0.625, p=0.007, and r=–0.762, p&lt;0.001, respectively). Conclusion: The levels of serum KL-6, and SP-D are elevated in SSc-ILD patients. Serum KL-6 may be a useful non-invasive biomarker for the disease severity, as determined by DLCO and the extent of fibrosis on HRCT, in patients with SSc-ILD. Trial registration: Thai Clinical Trials Registry, TCTR20200314001, registered 13 March 2020, retrospectively registered at http://www.thaiclinicaltrials.org/show/TCTR20200314001 Keywords: Scleroderma; Systemic sclerosis; Interstitial pneumonia; Interstitial lung disease; Fibrosis; Biomarker

Prediction of histopathological grades of liver pathology in cholestasis by direct bilirubin, aspartate aminotransferase, alanine aminotransferase and albumin level

Background: Liver biopsy, a routine diagnostic procedure in Dr. Soetomo General Hospital Surabaya, is a valuable tool in the diagnosis, prognosis, and management of parenchymal liver disease. However, the correlation between histopathological features and direct bilirubin, aspartate transaminase (AST), alanine transaminase (ALT) and albumin level has not yet been established. Objectives: To correlate various histopathological features with direct bilirubin, AST, ALT and albumin level of children with cholestasis. Method: This is a retrospective study of 51 cases of cholestasis diagnosed and treated from January 2011 to December 2016. All biopsies were reviewed and graded by a semi-quantitative scoring system according to Muthukanagarajan et al and categorized into fibrosis, bile duct proliferation, cholestasis and duct plate malformation. Degrees of all features were compared with direct bilirubin, AST, ALT, and albumin level. Statistical analysis used one way ANOVA, Kruskal-Wallis and unpaired t-test. p&lt;0.05 was considered significant. Results: There were 30 males and 21 females with a median age of 3 (1-9) months and a mean weight of 5 (1.41) kg. Degree of fibrosis was negative (47%), mild (22%), moderate (31%) and severe (0%). Bile duct proliferation was negative (57%), mild (21%), moderate (14%) and severe (8%). Cholestasis was negative in 0%, mild in 14%, moderate in 69% and severe in 18%; duct plate malformation was negative in 63% and positive in 37%. Direct bilirubin level showed significant difference with degree of duct proliferation (p=0.024). There was no significant difference of AST level with all degrees of histopathological grade. ALT level showed significant difference with degree of fibrosis (p=0.043). Albumin level showed significant difference with degree of fibrosis (p=0.000), degree of duct proliferation (p=0.006) and duct plate malformation (p=0.037). Conclusions: This study showed that while the direct bilirubin level was significantly associated with the degree of duct proliferation and the ALT level was significantly associated with the degree of fibrosis, the albumin level was significantly associated with the degree of fibrosis, degree of duct proliferation and degree of duct plate malformation. Sri Lanka Journal of Child Health, 2023: 52(2): 142-147

Neurointerventional Treatment of Vein of Galen Malformation (VGM): A Structured Review with a Proposal for the Comparison of Outcome Quality

Background: Vein of Galen malformation (VGM) is a congenital intracranial vascular anomaly consisting of arteriovenous fistulas and/or malformations between various arterial feeders and the median prosencephalic vein of Markowski (MPV). Despite its rare occurrence, VGM is of particular clinical relevance, as the excessive intracranial shunt volume leads to high mortality without appropriate treatment. Methods: The objective of this article is to review the published data on neurointerventional treatment and compare outcome quality in the included studies. Eight studies were included and synthesized. One study was multicentric and the rest were retrospective monocentric (level 4 evidence studies according to the Oxford Centre for Evidence-based Medicine). Results: The total number of included patients was 480 and patient age ranged from 1 day to 18 years. Mild or severe heart failure, hydrocephalus, and other reasons led to the indication for neurointerventional treatment, which was performed in all studies in the form of embolization. Under consideration of the introduced semiquantitative multidimensional scoring system, the highest total score, i.e., the best outcome quality, was found for the study “Houston” 2002–2018 (19 points) and the study “Duisburg” 2001–2010 (19 points). Conclusions: Neurointerventional treatment represents the essential pillar in the interdisciplinary management of patients with VGM, although standardization is lacking—based on the results of the structured review. As complementary treatments, pediatric critical care is mandatory and includes medical hemodynamic stabilization.

FAP expression in alpha cells of Langherhans insulae-implications for FAPI radiopharmaceuticals' use.

Radiopharmaceuticals targeting fibroblast activation protein (FAP) alpha are increasingly studied for diagnostic and therapeutic applications. We discovered FAP expression at immunohistochemistry (IHC) in the alpha cells of the Langerhans insulae of few patients. Therefore, we planned an investigation aimed at describing FAP expression in the pancreas and discussing the implications for radioligand applications. We retrospectively included 40 patients from 2 institutions (20 pts each) according to the following inclusion/exclusion criteria: (i) pathology proven pancreatic ductal adenocarcinoma and neuroendocrine tumors (NET), 10 pts per each group at each center; (ii) and availability of paraffin-embedded tissue; and (iii) clinical-pathological records. We performed IHC analysis and applied a semiquantitative visual scoring system (0, negative staining; 1, present in less than 30%; 2, present in more than 30% of the area). FAP expression was assessed according to histology-NET (n = 20) vs ductal adenocarcinoma (n = 20)-and to previous treatments within the adenocarcinoma group. The local ethics committee approved the study (No. INT 21/16, 28 January 2016). The population consisted of 24 males and 16 females, with a median age of 68 and a range of 14-84years; 8/20 adenocarcinoma patients received chemotherapy. In all the Langerhans insulae (40/40), pancreatic alpha cells were found to express FAP, with a score of 2. No difference was found among NET (20/20) and adenocarcinoma (20/20), nor according to neoadjuvant chemotherapy in the adenocarcinoma cohort (received or not received). Pancreatic Langerhans islet alpha cells normally express FAP. This is not expected to influence the diagnostic accuracy of FAP-targeting tracers. In the therapeutic setting, our results suggest the need to better elucidate FAPI radioligands' effects on the Langerhans insulae function.

Usefulness of 18F-Fluorodeoxyglucose Positron Emission Tomography in Diagnosing Polymyalgia Rheumatica and Large-Vessel Vasculitis: A Case-Control Study.

We aimed to evaluate the utility of FDG-PET/CT in diagnosing polymyalgia rheumatica (PMR) and associated large-vessel vasculitis (LVV). We analyzed FDG-PET/CT completed between 2015 and 2019 on patients diagnosed with PMR. For comparisons, patients with PMR were matched 1:1 to controls based on age and gender. FDG-PET/CT had been completed on the controls over the same period. The FDG uptake was scored visually for 17 articular or periarticular sites and 13 vascular sites using a semi-quantitative scoring system (score of 0-3). Eighty-one patients with PMR and eighty-one controls were included (mean age 70.7 (9.8) years; 44.4% women). Significant differences between the PMR and control groups were found at all articular and periarticular sites for the following: (i) the FDG uptake score (p < 0.001 for all locations); (ii) the number of patients per site with significant FDG uptake (score ≥ 2); (iii) the global FDG articular uptake scores (31 [IQR, 21 to 37] versus 6 [IQR, 3 to 10], p < 0.001); and (iv) the number of sites with significant FDG uptake (score ≥ 2) (scores of 0-17) (11 [IQR, 7 to 13] versus 1 [IQR, 0 to 2], p < 0.001). No significant differences in the global FDG vascular uptake scores were found between the patients who were considered isolated PMR and the control groups. The FDG uptake score and the number of sites with significant FDG uptake could be pertinent criteria for the diagnosis of PMR. Unlike others, we did not confirm the presence of vascular involvement in patients with isolated PMR.

What is the Effect of Bevacizumab on Cartilage and Synovium in a Rabbit Model of Hemophilic Arthropathy?

Hemophilic arthropathy can cause recurrent hemarthroses and severe damage to the synovium and articular cartilage. Previous studies have shown that vascular endothelial growth factor (VEGF) plays an essential role in neoangiogenesis. Bevacizumab, a monoclonal VEGF inhibitor, is used clinically to prevent angiogenesis. However, its effects on hemophilic arthropathy are unknown. Using a hemophilic arthropathy rabbit model, we asked: Does an intra-articular injection of bevacizumab (1) inhibit VEGF, (2) decrease signal intensity in dynamic contrast-enhanced MRI (DCE-MRI) as an assessment of capillary permeability and neoangiogenesis, (3) reduce cartilage damage, (4) reduce synovial changes, and (5) affect macroscopic changes during the development of hemophilic arthropathy? Twenty-five male New Zealand rabbits were divided into four groups. Eight knees from four rabbits were used as the control group. We used an established animal model for hemophilic arthropathy in the remaining 21 rabbits. Animals were assigned randomly to three groups with seven rabbits in each group. One group was used to establish mild arthropathy, and the other two were used to establish severe arthropathy. Autologous blood from the rabbits' ears was injected into the right and left knees twice per week for 8 weeks to represent mild arthropathy and for 16 weeks to represent severe arthropathy. In the mild arthropathy group, bevacizumab was injected into the right knee once every 2 weeks. Bevacizumab was injected into the right knee of rabbits in one of the severe arthropathy groups once every 2 weeks for 16 weeks, and intra-articular bevacizumab injections were administered to the right knees of rabbits in the other severe arthropathy group once every 2 weeks after the eighth week. An equal volume of 0.9% saline was injected into the left knee of rabbits in all arthropathy groups. To explore the efficacy of bevacizumab, joint diameters were quantitatively measured, and cartilage and synovial changes were examined. Degeneration of articular cartilage was evaluated with the semiquantitative Osteoarthritis Research Society International grading system. Synovial damage was analyzed with a semiquantitative microscopic scoring system. In addition, we evaluated perfusion and angiogenesis using DCE-MRI (quantitative signal intensity changes). Immunohistochemical testing was used to measure VEGF levels (analyzed by Western blotting). Intra-articular bevacizumab treatment inhibited VEGF in our rabbit model of hemophilic arthropathy. VEGF protein expression levels were lower in the mild arthropathy group that received intra-articular bevacizumab (0.89 ± 0.45) than the mild arthropathy control group (1.41 ± 0.61) (mean difference -0.52 [95% CI -0.898 to -0.143]; p = 0.02). VEGF levels were lower in the severe arthropathy group that received treatment for 16 weeks (0.94 ± 0.27) than in the control knees (1.49 ± 0.36) (mean difference -0.55 [95% CI -0.935 to -0.161]; p = 0.01). In the severe arthropathy group, the Osteoarthritis Research Society International score indicating cartilage damage was lower in the group that received intra-articular bevacizumab treatment from the beginning than in the control group (median 17 [range 13 to 18] versus 18 [range 17 to 20]; difference of medians 1; p = 0.02). Additionally, the scores indicated synovial damage was lower in the group that received intra-articular bevacizumab treatment from the beginning than the control group (median 5 [range 4 to 9] versus 9 [range 8 to 12]; difference of medians 4; p = 0.02). The mean of mean values for signal intensity changes was higher in the nontreated severe groups than in the group of healthy knees. The signal intensity changes were higher in the severe arthropathy control groups (Groups BC and CC) (median 311.6 [range 301.4 to 361.2] and 315.1 [range 269.7 to 460.4]) than in the mild arthropathy control group (Group AC) (median 234.1 [range 212.5 to 304.2]; difference of medians 77.5 and 81, respectively; p = 0.02 and p = 0.04, respectively). In the severe arthropathy group, discoloration caused by hemosiderin deposition in the cartilage and synovium was more pronounced than in the mild arthropathy group. In the severe arthropathy group treated with intra-articular bevacizumab, joint diameters were smaller than in the control group (Group BT median 12.7 mm [range 12.3 to 14.0] versus Group BC median 14.0 mm [range 13.1 to 14.5]; difference of medians 1.3 mm; p = 0.02). Hemarthrosis damages the synovial tissues and cartilage in the knees of rabbits, regardless of whether they are treated with intra-articular bevacizumab. However, intra-articular injection of bevacizumab may reduce cartilage and synovial damage in rabbits when treatment is initiated early during the development of hemophilic arthropathy. If the findings in this study are replicated in larger-animal models that consider the limitations of our work, then a trial in humans might be appropriate to ascertain whether intra-articular injection of bevacizumab could reduce cartilage damage and synovial changes in patients with hemophilia whose hemarthroses cannot otherwise be controlled.

Histological mapping of healing of the small and large intestine – A quantitative study in a porcine model

BackgroundGastrointestinal anastomoses are performed in many patients every year. The pathogenesis of aberrant anastomotic healing and the causes of intestinal leakage are not fully understood. The present study gathered and critically evaluated histological quantitative data to deepen current knowledge of anastomotic healing in the small and large intestine and its complications and outline the options for further experimental in vivo research in large porcine animal models. MethodsThree groups of porcine intestinal anastomoses were compared: small intestine without defect (SI; n = 7), small intestine with an additional defect (SID; n = 8), and large intestine (LI; n = 7). Multilevel sampling (2112 micrographs) and stereological methods were used for histological quantification of proliferation (Ki-67 immunohistochemistry), neutrophil infiltration (myeloperoxidase staining), vascularity (von Willebrand factor) and type I and type III collagen formation (picrosirius red in polarized light) within the region of anastomosis compared to the region outside of anastomosis. ResultsQuantitative histological evaluation revealed the following results. i) Proliferation, vascularity, and collagen, but not neutrophils, were more highly expressed within the anastomosis than outside of the anastomosis region. ii) Porcine large and small intestine were not interchangeable based on histological evaluation of surgical experiments. The presence or absence of an additional experimental defect strongly affected healing, but the healing seemed complete after 21 days. iii) The microscopic structure of small intestine segments was more affected by their proximity to the anastomosis than the structure of large intestine segments. ConclusionsHistological quantification was more laborious than the previously used semiquantitative scoring system evaluating the healing rate of intestinal anastomoses, but it provided detailed maps of biological processes within individual intestine layers. The primary data collected in the study are open and available for power sample analyses to calculate the minimum numbers of samples justified in future experiments on porcine intestines. The porcine intestine is a promising animal model with translational potential for human surgery.

ULTRASONOGRAPHIC EVALUATION OF COELOMIC FLUID IN HEALTHY CORN SNAKES (PANTHEROPHIS GUTTATUS).

Free coelomic fluid can be a physiologic or pathologic finding in snakes. In this study, ultrasonography was used to assess the presence, volume, and type of coelomic fluid in 18 (16 females, 2 males) clinically healthy corn snakes (Pantherophis guttatus) using a semiquantitative scoring system. Each snake was divided into five equal regions (R1-R5) by length (from rostrum to vent) and fluid volume was scored on a scale of 0 to 4. Coelomic fluid was sampled when possible and cytologic analysis was performed. The majority of snakes (16/18) were found to have some degree of free coelomic fluid. The coelomic fluid samples that were collected (n= 6) were classified as transudates, acellular or primarily lymphocytic. Fluid was most likely to be found in R3 compared with all other regions and least likely to be found in R1 compared with R2, R3, and R4. A higher score of volume was found in R3 compared with R1 and R5. This study describes the distribution and abundance of coelomic fluid in snakes, as well as a method for point of care ultrasound (POCUS) in this species.

Development and Validation of a Multiparametric Semiquantitative Scoring System for the Histopathological Assessment of Ischaemia Severity in Skeletal Muscle

Skeletal muscle is one of the most abundant and dynamic tissues of the body, with a strong regenerative capacity. Muscle injuries can occur as a result of a variety of events, including tissue ischaemia. Lower limb ischaemia occurs when there is an insufficient nutrient and oxygen supply, often caused by stenosis of the arteries due to atherosclerosis. The aim of this study was to develop and validate a multiparametric scoring tool for assessing ischaemia severity in skeletal muscle in a commonly used preclinical animal model. Tissue ischaemia was surgically induced in mice by ligation and excision of the femoral artery. Calf muscles were carefully dissected, prepared for histological analysis, and scored for inflammation, fibrosis, necrosis, adipocyte infiltration, and muscle fibre degeneration/regeneration. Kendall’s coefficient of concordance (W) showed a very good agreement between the appraisers when scoring each individual histological feature: inflammation (W = 0.92, p ≤ 0.001 ), fibrosis (W = 0.94, p ≤ 0.001 ), necrosis (W = 0.77, p ≤ 0.001 ), adipocyte infiltration (W = 0.91, p ≤ 0.001 ), and fibre degeneration/regeneration (W = 0.86, p ≤ 0.001 ). Intrarater agreement was also excellent (W = 0.94 or more, p ≤ 0.001 ). There was a statistically significant negative association between the level of muscle ischaemia damage and the calf muscle weight and skeletal muscle fibre diameter. Here, we have developed and validated a new multiparametric, semiquantitative scoring system for assessing skeletal muscle damage due to ischaemia, with excellent inter- and intrarater reproducibility. This scoring system can be used for assessing treatment efficacy in preclinical models of hind limb ischaemia.

Musculoskeletal Ultrasound in Monitoring the Efficacy of Gout: A Prospective Study Based on Tophus and Double Contour Sign

In patients with gout receiving uric acid-lowering therapy, musculoskeletal ultrasound has the potential to observe changes in gout lesions. To analyze the effectiveness of uric acid-lowering therapy in patients with gout over one year using musculoskeletal ultrasound as a monitoring technique. Prospective cohort study. A total of 215 patients meeting the 1977 American College of Rheumatology gout classification criteria and treated with uric acid-lowering therapy were separated into two groups, treat-to-target and treat-to-non-target depending on the target serum urate levels. Lower extremity joints were evaluated by ultrasound before therapy (M0), as well as three (M3), six (M6), and twelve (M12) months after therapy. At various moments during uric acid-lowering therapy, the tophus size and the semiquantitative ultrasound scoring system of double contour sign were measured in the treat-to-target and treat-to-non-target groups. Ninety-five tophi (45 in treat-to-target and 50 in treat-to-non-target) and sixty-seven double contour sign (34 in treat-to-target and 33 in treat-to-non-target) were evaluated longitudinally. In both groups, the long diameter, short diameter, and area of tophus in treat-to-target decreased as the duration of uric acid-lowering treatment increased. Differences in the long diameter of tophus between M12 and M0, M3 and M6 were statistically significant (P < 0.05), while differences between the other time points were not significant (P > 0.05). No statistically significant differences were observed in the short diameter and the area of tophus between M0 and M3 (P > 0.05), while there were statistically significant differences between other periods (P < 0.05). In treat-to-non-target, the long diameter, short diameter, and area of tophus showed a slight increase at different uric acid-lowering therapy time points. The differences in the long diameter, short diameter, and area of tophus at different uric acid-lowering therapy time points were not significant (P > 0.05). The semiquantitative ultrasound scoring system of double contour sign of treat-to-target and treat-to-non-target showed a decreasing trend with increasing uric acid-lowering therapy time, with a more pronounced drop in treat-to-target than treat-to-non-target. In treat-to-target, the difference in the semiquantitative ultrasound scoring system of double contour sign at each uric acidlowering therapy time point was significant (P < 0.05). In treat-tonon- target, the difference in semiquantitative ultrasound scoring system of double contour sign scores between M0 and M3 was not statistically significant (P >0.05), but it was statistically significant for the remaining time points (P < 0.05). After one year of uric acid-lowering therapy in patients with gout, an ultrasound indicated that the size of tophus and the semiquantitative ultrasound scoring system of double contour sign score decreased dramatically in the treat-to-target group. Semiquantitative ultrasound scoring system of double contour sign score was dramatically reduced in the treat-to-non-target group, but the size of the tophus remained the same. Therefore, musculoskeletal ultrasound is an effective tool to monitor the efficacy of uric acid-lowering therapy.

Damage analysis of retrieved BioloxⓇdelta components used in hard and soft bearings.

This retrieval study included 43 Biolox delta explants (18 CoC, 25 CoP). Implants were examined macroscopically, whereby damage was evaluated using a semi quantitative scoring system. Confocal microscopy was used to examine wear related damage patterns of the articulating surfaces. Scanning electron microscopy (SEM) with energy-dispersive X-ray spectroscopy (EDS) was used to analyze wear marks on the implant surface and wear debris in periprosthetic tissue samples. Raman spectroscopy and X-ray diffraction (XRD) were used to quantify monoclinic zirconia fractions. On all components, in vivo wear resulted predominantly in different damage patterns caused by metal transfer. In CoC bearings stripe wear was additionally detected, and some implants underwent severe damage due to component breakage. The wear scores were higher for CoC components, with no differences between the scores for CoC heads and liners. Wear features caused comparable roughening on implants from CoC and CoP bearings. SEM studies demonstrated that most wear marks were caused by metal debris released from implant components. Grain pull-out was observed in stripe wear regions. Monoclinic phase shift was observed in a similar quantity on components from CoP and CoC bearings. The increase of monoclinic zirconia content around metal deposits was minimal and was more pronounced in areas of stripe wear. The results of this study indicate, that ZTA components in general undergo minimal wear in both, CoC and CoP bearings, however, it is more pronounced in the former. Metal deposits, as the most common wear feature, have no significant effect on monoclinic phase transition. STATEMENT OF SIGNIFICANCE: In this paper, we classify all damage patterns macroscopically according to an established scoring system and assess them regarding surface roughness (confocal microscopy) and monoclinic phase content (Raman spectroscopy) in order to derive the severity for patients. We compare hard-hard and hard-soft bearings and relate damage patterns with metal transfer based on SEM/EDS examinations. Furthermore, we work out correlations between patient-specific data, cause of revision and the physical condition of each individual sample Our cohort consists of 43 Biolox delta retrievals, a comparatively large quantity. In addition, we address current topics such as metal transfer and, based on the classification of damage patterns, provide incentives and/or meaningful focal points for further research.

An Integrated Analysis Reveals Ciliary Abnormalities in Antrochoanal Polyps.

The mechanisms underlying the antrochoanal polyps (ACPs) remained unclear. We aimed to identify the differentially expressed genes (DEGs) profile, the cilia-related genes expression levels and the morphological characteristics of ciliated cells in patients with ACPs. We obtained ACPs biopsy samples from 28 patients and uncinate process from 27 healthy controls. Whole-transcriptome RNA sequencing, immunofluorescence staining, quantitative polymerase chain reaction, and scanning electron microscopy were performed. 3739 DEGs were detected between ACPs and controls, and Gene Ontology analysis on these DEGs implicated cilium assembly, cilium motility, cilia component, cilia function, inflammatory response and immune system process were included in ACPs pathogenesis. Gene set enrichment analysis implicated sets of genes regulated in processes associated with cilium organization, cilium morphogenesis, cilium movement, axoneme assembly, axonemal dynein complex assembly and cell projection assembly. The expression levels of cilia-related genes (FOXJ1, DNAI1, DNAH9, RSPH1, RSPH9 and RSPH4A) were validated by quantitative polymerase chain reaction (Fold change >2, P<0.05) and FOXJ1 was positive correlated with DNAI1, DNAH9, RSPH4, RSPH1, RSPH9, DNAH5, DNALI1 in ACPs (all P < 0.05). Based on our semi-quantitative scoring system, median scores of α-Tubulin, DNAI1 and RSPH4A were significantly higher in ACPs than in controls. In addition, loss of ciliated cells and a shorter cilia pattern were further confirmed by immunofluorescence staining and scanning electron microscopy in ACPs. The aberrant expression of cilia-related genes and ciliary structural impairment are an important pathological phenomenon in ACPs, and our findings may provide novel insights into understanding the mysterious mechanisms underlying ACPs.

Abstract 5: Malignant Venous Outflow Profile Predicts Futile Recanalization After Endovascular Thrombectomy.

Objective: To explore the association of baseline venous outflow from the occluded cerebral territory and futile recanalization (FR) in patients with acute ischemic stroke due to large vessel occlusion (LVO) in the anterior circulation. Methods: Consecutive patients from a prospectively maintained stroke database were included in this study if they presented with a multimodal CT-confirmed LVO in the anterior circulation and underwent thrombectomy within 24 hours after stroke onset. Excellent recanalization of the occluded artery, defined as final mTICI 2c-3 on digital subtraction angiography, was considered to be futile if patients fail to achieve functional independence (modified Rankin Scale of 0-2) at 90 days. Baseline venous outflow profile was determined by a 7-point semi-quantitative scoring system which assessed ipsilateral venous opacification of the vein of Labbe, Trolard and superficial middle cerebral vein on single-phase CTA source images. Hypoperfusion intensity ratio (HIR, defined as delay time &gt;6 seconds/delay time &gt;3 seconds) was applied to evaluate tissue-level collateral status. Results: 145 patients met the inclusion criteria. Multivariable logistic regression analysis showed that malignant venous outflow (OR 4.30 [95% CI 1.41-14.57]; p =0.013) was independently associated with functional dependence regardless of age, glucose, HIR, early neurologic deterioration, hemorrhagic transformation and final infarct volume. FR was observed in 24/52 (46%) recanalized patients. Stepwise logistic regression revealed a poor venous outflow profile at presentation as an independent predictor of FR (OR 7.04 [95% CI 1.29-50.16]; p =0.03) adjusted for baseline NIH Stroke Scale score, HIR and presence of hemorrhagic transformation. Conclusion: A malignant venous outflow profile at baseline is associated with FR in patients with LVO. The mechanism by which poor venous outflow from the occluded territory hinders clinical recovery despite prompt successful thrombectomy remains to be determined.

Gout of feet and ankles in different stages: The potentiality of a new semiquantitative DECT scoring system in monitoring urate deposition.

The purpose of this study was to investigate the diagnostic performance of a novel semi-quantitative dual-energy computed tomography (DECT) scoring system in monitoring urate deposition. This study included 287 patients with gout. All patients underwent ankle/foot DECT scans. DECT scores at different stages were compared and their diagnostic efficacies evaluated. Associations between DECT scores and clinical variables were evaluated. Gouts with positive DECT results in early, middle, and late stages were 78.5 %, 81.4 %, and 95.8 %, respectively (all P > .05). The total and ankle/midfoot DECT scores at different stages significantly increased with disease duration (all P < .05). DECT scores of 4 regions excluding the first metatarsophalangeal joint in early and middle stages were lower than those in late stage (all P < .05). DECT scores achieved excellent diagnostic performance for differentiating gout in early stage from middle and late stages (area under the curve, 0.923 and 0.949), with high sensitivity, specificity, positive predictive value, and negative predictive value (all > 85 %). Total DECT score was highly positively correlated with the volume of urate crystals (R = 0.873, P < .001). Disease duration, serum uric acid level, bone erosion, and Achilles tendon involvement significantly affected total DECT scores (all P < .01). In conclusion, longer disease duration, higher serum uric acid levels, bone erosion, and Achilles tendon involvement were closely associated with total DECT scores. DECT scoring system may be an invaluable tool for gout diagnosis owing to its high detection efficacy and a surrogate method to evaluate the amount of urate crystals and erosion of surrounding tissues.

Clinical value of the renal pathologic scoring system in complement-mediated thrombotic microangiopathy

Objectives This study was initiated to establish a renal thrombotic microangiopathy (TMA) scoring system based on clinical needs and investigate its predictive value for patients’ long-term outcomes. Methods Kidney biopsy-proven Complement-mediated TMA (C-TMA) patients from January 2000 to December 2017 in Peking University First Hospital were retrospectively studied. Both acute and chronic TMA-related lesions, including 15 pathologic indices, were semiquantitatively scored. The interobserver and intraobserver reproducibility and correlation between the pathologic indices and clinical parameters were analyzed. Furthermore, the patients were divided into 2 groups by dialysis use at baseline, and the association of these pathologic indices with their prognostic outcomes was assessed between the two groups. Results Ninety-two patients with renal biopsy-proven C-TMA were enrolled. All fifteen included pathology indices showed good or moderate interobserver and intraobserver reproducibility and correlated well with several clinical parameters. Several clinicopathological indices were worse in the dialysis group than in the nondialysis group, such as serum creatinine, hemoglobin, platelet count, and estimated glomerular filtration rate. Moreover, morphologic features in the dialysis group presented with more severe vascular lesions. Interstitial fibrosis and chronic tubulointerstitial lesions were related to a trend of high risk of continuous dialysis in the dialysis group. Based on univariate and multivariable Cox regression analysis, more severe glomerular lesions, including glomerular mesangiolysis, glomerular basement membrane double contours and glomerular mesangial proliferation, were identified as risk factors predicting worse prognosis. Conclusions Our renal C-TMA semiquantitative scoring system is reliable with good reproducibility and prognostic value in clinical practice, which needs further validation.

Survivin and p53 expression in benign and malignant lesions of breast

Breast cancer is the most common malignancy in women, accounting for 14 % of all cancers in India; it accounts for &amp;#60; 1% of all cancer cases in men. Breast cancer is the leading cause of death worldwide. The study aims to assess the expression of p53 and Survivin in Fibroadenoma and Invasive carcinoma of no special type and its prognostic importance.A total number of 75 cases are included in the study, out of which 34 are reported by two histopathologists as invasive carcinoma NST (ductal) and 41 reported as Fibroadenoma. All the cases included in the study are females. Immunohistochemistry for p53 and Survivin performed in both the groups with positive and negative controls. Cytoplasmic or nuclear IHC expression for Survivin is considered to be positive. Whereas, nuclear positivity is considered for p53. A semi quantitative scoring system performed consisting of intensity and % of positive cells. The expression of Survivin and p53 is found to be high in IDC, 79% and 85% respectively. The correlation is found to be significant (p value = 0.002, Spearman Rho correlation). However, the expression of Survivin is seen in one third of all the cases of Fibroadenoma (37.5%) and p53 expression is expectedly low, seen in 12% cases. Over expression of Survivin in IDC patients compared to benign cases is found to be correlated with p53 expression. Though Survivin is likely to contribute to apoptosis resistance, its expression is found to confer resistance to chemotherapy and radiotherapy in IDC. Henceforth, Survivin over expression along with p53 in invasive breast carcinoma defining overall poor prognosis and low survival rates.

Shorter Cilia Length and Aberrant Ciliated Marker DNAI1 in Allergic Rhinitis.

This study aimed to investigate whether the impaired ciliary length and aberrant ciliary ultrastructure marker, dynein axonemal intermediate chain 1 (DNAI1), are important pathological characteristics in nasal mucosa from patients with allergic rhinitis (AR). Biopsies were taken from the inferior turbinate (IT) of controls (n = 20) and patients with AR (n = 20). The ciliary length and the DNAI1 location patterns were assessed by using immunofluorescent staining. Three patterns of DNAI1 localization were defined using a semi-quantitative scoring system: normal (N), partial (P) and absence (A). Every individual section was assigned a score between 0 and 2 in each high-power field (5 fields per sample). The score of 0 = pattern N >70%; 1 = patterns N + P >70%; and 2 = pattern A ≥30%. The receiver operating characteristic (ROC) curve was used to evaluate the predicted value of DNAI1 score for AR. The ciliary length was reduced by 33.3% in patients with AR compared with controls (P < 0.0001). The higher DNAI1 score was found in the AR group, with a median (first and third quartile) of 0.9 (0.4 and 1.08), which was 0.1 (0 and 0.76) in the control group (P = 0.0071). The ROC of DNAI1 was calculated based on the area under the curve of 0.74 (P = 0.0094). The cutoff value of ROC was 0.5833, with a sensitivity and specificity of 70%. These results suggested that the shorter ciliary length and aberrant localization of DNAI1 are potentially important pathological characteristics of the allergic nasal mucosa. The aberrant localization of DNAI1 may provide a novel candidate target for clinical management of AR.