Semiquantitative Grading Scale Research Articles

High-resolution peripheral quantitative computed tomography for the assessment of acro-osteolysis and calcinosis in patients with systemic sclerosis

ObjectiveTo assist the development of future treatments in systemic sclerosis (SSc), the development of reliable outcome measures is pivotal. We aimed to evaluate the use of high-resolution peripheral quantitative CT (HR-pQCT) for visualization and gradation of acro-osteolysis (AO) and calcinosis compared to conventional hand radiographs (CR) in patients with SSc. MethodsHR-pQCT scans of the 2nd to 4th fingers, CR, nail fold capillaroscopy, and a clinical examination were conducted. Images were reviewed for the presence and degree of AO and calcinosis according to semiquantitative grading scales. ResultsForty patients were included. Fourteen had AO according to CR, whereas HR-pQCT revealed AO in 18 patients. The sensitivity and specificity of classifying patients as having AO by HR-pQCT when CR was used as reference were 93% (95% CI: 66–99%) and 80% (95% CI: 59–93%), respectively. By CR and with HR-pQCT as reference, the sensitivity and specificity were 72% (95% CI: 47–90%) and 95% (95% CI: 76–99%). Patients with AO had more or larger calcifications than patients without AO according to the proposed HR-pQCT grading system, with a median grade of 2 (IQR: 1–3) versus 0 (IQR: 0–1) (P<0.01). Grade 3 changes were observed exclusively in patients with AO (n=6/14, 42.9%). Assessment of AO and calcinosis by HR-pQCT demonstrated moderate to excellent test-retest reliability. ConclusionHR-pQCT allowed precise and reliable classification and grading of acro-osteolysis and acral calcinosis. The modality could prove helpful for detecting and monitoring these lesions as well as facilitating early diagnosis and guide treatment of these patients.

B-029 Validation of an Automated Analyzer for Urine Microscopic Examination

Abstract Background Microscopic examination of urine sediment can be time-consuming and labor intensive. Automated analyzers can help to standardize resulting and reduce manual work. Urinalysis has traditionally utilized semi-quantitative grading scales, eg. “0–3/high power field”. The Sysmex UF 5000 (Symex, Kobe, JP) uses flow cytometry to enumerate microscopic particles including white and red blood cells (WBC, RBC), bacteria, casts, and epithelial cells (EC). We sought to validate the counting of these microscopic particles against our reference manual method, and also determine if the research parameters from the analyzer could be used to accurately measure other particles in urine (eg. crystals). Methods Residual urine samples were analyzed for microscopic particles both by the UF 5000 and manually using KOVA tubes (KOVA International, Garden Grove, CA). Particles were enumerated using the laboratory’s current semi-quantitative grading system. Exact agreement and agreement +/- one level were calculated. Results Conclusion The analyzer results were comparable to manual resulting; in discrepant cases, the UF result was typically higher than manual, which may reflect the increased sensitivity of this method. Additional studies are ongoing to determine if the UF 5000 research parameters are accurate for semi-quantitative reporting of pathologic casts, crystals, and yeast.

Oxidative stress status assessment of rats' brains injury following subacute exposure to K-oximes

Oxidative stress status and morphological injuries in the brain of Wistar rats induced by repeated application of selected acetylcholinesterase reactivators – asoxime, obidoxime, K027, K048, K074, and K075 were evaluated. Each oxime in a dose of 0.1 of LD50/kg im was given 2x/week for 4 weeks. Markers of lipid peroxidation (malondialdehyde, MDA), and protein oxidation (advanced oxidation protein products, AOPP), as well as the activity of antioxidant enzymes (catalase, CAT, superoxide dismutase, SOD, glutathione reductase, GR, and glutathione peroxidase, GPx), were estimated in the brain tissue homogenates on day 35 of the study. Brain alterations were carefully quantified by semiquantitative grading scales – brain damage score (BDS). Oxidative stress parameters, MDA and AOPP were significantly highest in the asoxime-, obidoxime- and K075-treated groups (p < 0.001). The activity of SOD and CAT was significantly elevated in the obidoxime-, K048-, and K075-treated groups (p < 0.001). Besides, GR was markedly decreased in the obidoxime- and K074-treated groups (p < 0.01), while treatment with K048, K074 and K075 induced extremely high elevation in GPx levels (p < 0.001). In the same groups of rats, brain alterations associated with polymorphonuclear cell infiltrate were significantly more severe than those observed in animals receiving only asoxime or K027 (p < 0.001). The presented results confirmed that treatment with different oximes significantly improved the oxidative status and attenuated signs of inflammation in rats’ brains. Presented results, together with our previously published data can help to predict likely adverse systemic toxic effects, and target organ systems, which are crucial for establishing risk categories, as well as in dose selection of K-oximes as drug candidates.

Prognostic factors affecting structural integrity after arthroscopic rotator cuff repair: a clinical and histological study.

Tendon degeneration contributes to rotator cuff tears; however, its role in postoperative structural integrity is poorly understood. The purpose of this study was to investigate the factors associated with postoperative structural integrity after rotator cuff repair, particularly focusing on the histology of tendons harvested intraoperatively. A total of 56 patients who underwent primary arthroscopic rotator cuff repair between 2009 and 2011 were analyzed. A 3-mm-diameter sample of supraspinatus tendons was harvested en bloc from each patient after minimal debridement of the torn ends. Tendon degeneration was assessed using seven histological parameters on a semi-quantitative grading scale, and the total degeneration score was calculated. One-year postoperative magnetic resonance imaging was used to classify the patients based on retear. The total degeneration scores in the healed and retear groups were 13.93±2.03 and 14.08±2.23 (P=0.960), respectively. Arthroscopically measured anteroposterior (AP) tear sizes in the healed and retear groups were 24.30±12.35 mm and 36.42±25.23 mm (P=0.026), respectively. Preoperative visual analog scale pain scores at rest in the healed and retear groups were 3.54±2.37 and 5.16±2.16 (P=0.046), respectively. Retraction sizes in the healed and retear groups were 16.02±7.587 mm and 22.33±13.364 mm (P=0.037), respectively. The odds of retear rose by 4.2% for every 1-mm increase in AP tear size (P=0.032). The postoperative structural integrity of the rotator cuff tendon was not affected by tendon degeneration, whereas the arthroscopically measured AP tear size of the rotator cuff tendon was an independent predictor of retear.

The Value of Superb Microvascular Imaging and Contrast-enhanced Ultrasound for the Evaluation of Neovascularization in Carotid Artery Plaques

To compare the consistency between superb microvascular imaging (SMI) and contrast-enhanced ultrasound (CEUS) for the detection of neovascularization in carotid plaques of different thicknesses and to evaluate the applied value of these two methods for detecting neovascularization in carotid plaques in the clinic. A total of 45 patients with carotid artery plaques who were diagnosed in our hospital involving 76 hypoechoic plaques with a thickness ≥2.0 mm were selected. According to thickness, the plaques were divided into three groups: 2.0-2.5 mm, 2.5-3.0 mm and ≥3.0 mm. Each group underwent both SMI and CEUS, and two experienced sonographers (A and B) analyzed the ultrasound images to evaluate the neovascularization of carotid plaques. The amount of the neovascular signal was assessed using a semi-quantitative grading scale (vascularity grade: grade 0-3). SMI and CEUS were graded respectively according to the visual methods as follows: grade 0: no blood flow signal/enhancement within plaques; grade 1: a few blood flow signals/enhancement within plaques; grade 2: medium blood flow signals/enhancement within plaques; and grade 3: extensive blood flow signals/enhancement within plaques. Kappa consistency test was used to analyze the consistency of the grade of neovascularization in plaques between SMI with CEUS. Gamma rank correlation analysis was used to examine the correlation between neovascularization grade by SMI and CEUS in plaque and plaque thickness. Of these patients, 14 had unilateral plaques and 31 had bilateral plaques. The two sonographers were highly consistent in terms of applying SMI and CEUS methods for diagnosing neovascularization in carotid plaques (Kappa values were 0.736 and 0.680>0). Consistency was found between SMI and CEUS by sonographers (sonographer A: Kappa=0.823; sonographer B: Kappa=0.842) in evaluating the neovascular grade in the carotid plaques. SMI and CEUS grades were positively correlated with plaque thickness (sonographer A: γ=0.735 and 0.772; sonographer B: γ=0.805 and 0.798). Neovascularization in carotid plaques was successfully detected by SMI in a manner that concurred well with CEUS results. Our data indicate that both CEUS and SMI have high diagnostic value for assessing the neovascularization of plaques.

MO264: Selective Endothelin a Receptor Antagonist Atrasentan Attenuates Mesangial Cell Injury, Proteinuria and intra-renal proliferative, inflammatory and Fibrotic Transcriptional Networks in a Rat Model of Mesangioproliferative Glomerulonephritis

Abstract BACKGROUND AND AIMS Mesangial cell (MC) activation, characterized by excess proliferation and extra-cellular matrix deposition is the initiating intra-renal event in the pathogenesis of mesangioproliferative glomerulonephritis (MPGN), including IgA nephropathy (IgAN). However, the molecular mechanisms responsible for MC activation and the subsequent podocyte crosstalk resulting in proteinuria have not been well defined. The objective of this study was to determine the role of the endothelin A (ETA) receptor in MC activation, proteinuria and pathogenic intra-renal transcriptional responses in a rat model of MPGN, using atrasentan, a potent and selective ETA antagonist. METHOD MPGN was induced in male Wistar rats by a single IV injection of anti-Thy1.1 antibody (0.5 mg/kg) on day 0. Rats were randomized to receive atrasentan (10 mg/kg, po bid) or vehicle control beginning on Day 1 through Day 7. UPCR was measured on Days 1, 3, 5 and 7; on Day 7, kidneys were fixed, stained and evaluated by light microscopy using a semi-quantitative grading scale by a pathologist, blinded to treatment allocations. RNA-seq was performed on flash-frozen kidney cortex and analyzed using DESeq2 to identify differentially expressed genes followed by gene set enrichment analysis to identify dysregulated transcriptional networks that were cross-validated to the glomerular transcriptome of kidney biopsy samples from IgAN patients (GSE104066). Cell type-specific signatures were derived from a scRNA-seq dataset (GSE171314) from IgAN patients. RESULTS Glomerular injury was characterized histologically by prominent mesangial hypercellularity and matrix expansion, glomerular adhesions, segmental mesangiolysis and glomerulosclerosis, and was significantly attenuated by atrasentan. Tubular injury, including protein casts, tubular degeneration, tubular dilation and interstitial fibrosis, was also observed in the model, although more modest and likely secondary to the glomerular injury, and was also significantly attenuated by atrasentan. The targeted mesangial injury induced by anti-Thy1.1 also resulted in significant proteinuria, which was substantially reduced by atrasentan (Fig. 1). In addition, atrasentan significantly attenuated the increase in kidney weight, likely reflective of the reduced mesangial proliferation, matrix expansion and accompanying inflammation observed histologically. Induction of MPGN in rats was transcriptionally characterized by a downregulation of metabolism gene networks and upregulation of networks associated with proliferation, inflammation and fibrosis, consistent with the hallmark genesets dysregulated in the glomeruli of IgAN patients (Fig. 2). Atrasentan downregulated these intra-renal proliferative, inflammatory and fibrotic transcriptional networks and upregulated metabolism networks (Fig. 2). Analysis using cell-type specific signatures derived from scRNA-seq data from IgAN patients, suggests that atrasentan treatment attenuates anti-Thy1.1-induced gene expression changes in multiple cell types, including mesangial cells, the target cell of injury in MPGN. In addition, although anti-Thy1.1 primarily induces glomerular injury, there was a strong transcriptional signature of failed repair in the proximal tubules, which was attenuated by atrasentan. CONCLUSION This study suggests an important role of the ETA receptor in MC activation, subsequent proteinuria and activation of pathogenic proliferative, inflammatory and fibrotic intra-renal transcriptional networks in MPGN. This further supports the therapeutic potential of atrasentan, a selective ETA receptor antagonist, to attenuate mesangial cell activation, proteinuria and pathogenic intra-renal signaling in MPGNs such as IgAN.

Neutrophil-to-lymphocyte ratio is associated with carotid intraplaque neovascularization in asymptomatic carotid stenosis patients.

Neutrophil-to-lymphocyte ratio (NLR) has been suggested as an available systemic inflammatory biomarker. This study aims to evaluate whether intraplaque neovascularization assessed by contrast-enhanced ultrasound (CEUS) is associated with NLR in asymptomatic carotid stenosis patients. One hundred and forty-four asymptomatic patients with carotid luminal stenosis >30% were assessed using contrast-enhanced ultrasound imaging. The contrast enhancement within the plaque was classified on a visual semiquantitative grading scale. The data collected included the patient's risk factors, laboratory results, cardiovascular disease history, and drug use history. Univariate and multivariate analyses were assessed to identify independent factors related to intraplaque neovascularization with adjustment for potential confounders. Patients with CEUS grade 2 plaques had a higher level of LDL-C (p < .001), neutrophil count (p < .001), and blood glucose (p=.005), but lower level of lymphocyte count (p=.021). The presence of grade 2 plaques was significantly associated with high NLR values (OR 1.21, 95% CI 1.03-1.43, p=.017). Patients were divided into four groups according to the quartile of NLR values. Compared to the patients in the first quartile of NLR (<1.73), the patients in the fourth NLR quartile (≥3.38) were characterized by the most prevalence of CEUS grade 2 plaques (OR 4.55, 95% CI 1.69-12.25, p=.003). Multivariate logistic regression analysis after adjusting various variables demonstrated NLR remained an independent risk factor for the presence of CEUS grade 2 plaques. Intraplaque neovascularization is significantly associated with NLR in asymptomatic carotid stenosis patients.

Developing a grading scale for the evaluation of degenerative changes in uncovertebral (Luschka) joints.

The study aimed to develop a grading scale for evaluating degenerative changes in uncovertebral joints (UVJs) found on computed tomography (CT) scans, and assess the intra- and inter-rater reliability of this scale. The study included 50 subjects (average age 55.12 ± 13.76 years) referred for CT examinations due to cervical complaints. Three researchers developed a reading protocol using a semiquantitative grading scale of UVJ degenerative changes. CTs were read and reread several times to refine the protocol. Subsequently, the reader read and reread 20 CTs (C3-C7 bilateral, altogether 200 UVJs) 2 weeks apart in order to assess the intra-rater reliability. The second reader, after the training, read the same 20 CTs (yet, C4-C6 bilateral, altogether 120 UVJs) in order to assess the inter-rater reliability. The additional 30 CTs were evaluated to analyze the prevalence and associations between the studied parameters. The final grading scale included joint space narrowing, osteophyte, subchondral sclerosis, subchondral cyst, hypertrophy, cortical erosion, and vacuum phenomenon. The intra-rater agreement ranged from very good in joint space narrowing (k= 0.757), osteophyte grade (k= 0.748), subchondral cyst (k= 0.716), hypertrophy (k= 0.728), vacuum phenomenon (k= 0.712) to good (k= 0.544) in subchondral sclerosis, and fair in cortical erosion (k= 0.36). For the overall UVJ degeneration grade, the intra-rater agreement was very good (k= 0.616) and the inter-rater reliability was good (k= 0.468). 77.6% of the evaluated UVJs exhibited some degenerative changes. Our grading scale is the first to evaluate UVJs presenting good-very good intra-rater and inter-rater reliability which can be utilized in further clinical studies and assess degenerative changes in UVJs.

Quantitative Muscle MRI in Patients with Neuromuscular Diseases-Association of Muscle Proton Density Fat Fraction with Semi-Quantitative Grading of Fatty Infiltration and Muscle Strength at the Thigh Region.

(1) Background and Purpose: The skeletal muscles of patients suffering from neuromuscular diseases (NMD) are affected by atrophy, hypertrophy, fatty infiltration, and edematous changes. Magnetic resonance imaging (MRI) is an important tool for diagnosis and monitoring. Concerning fatty infiltration, T1-weighted or T2-weighted DIXON turbo spin echo (TSE) sequences enable a qualitative assessment of muscle involvement. To achieve higher comparability, semi-quantitative grading scales, such as the four-point Mercuri scale, are commonly applied. However, the evaluation remains investigator-dependent. Therefore, effort is being invested to develop quantitative MRI techniques for determination of imaging markers such as the proton density fat fraction (PDFF). The present work aims to assess the diagnostic value of PDFF in correlation to Mercuri grading and clinically determined muscle strength in patients with myotonic dystrophy type 2 (DM2), limb girdle muscular dystrophy type 2A (LGMD2A), and adult Pompe disease. (2) Methods: T2-weighted two-dimensional (2D) DIXON TSE and chemical shift encoding-based water-fat MRI were acquired in 13 patients (DM2: n = 5; LGMD2A: n = 5; Pompe disease: n = 3). Nine different thigh muscles were rated in all patients according to the Mercuri grading and segmented to extract PDFF values. Muscle strength was assessed according to the British Medical Research Council (BMRC) scale. For correlation analyses between Mercuri grading, muscle strength, and PDFF, the Spearman correlation coefficient (rs) was computed. (3) Results: Mean PDFF values ranged from 7% to 37% in adults with Pompe disease and DM2 and up to 79% in LGMD2A patients. In all three groups, a strong correlation of the Mercuri grading and PDFF values was observed for almost all muscles (rs > 0.70, p < 0.05). PDFF values correlated significantly to muscle strength for muscle groups responsible for knee flexion (rs = −0.80, p < 0.01). (4) Conclusion: In the small, investigated patient cohort, PDFF offers similar diagnostic precision as the clinically established Mercuri grading. Based on these preliminary data, PDFF could be further considered as an MRI-based biomarker in the assessment of fatty infiltration of muscle tissue in NMD. Further studies with larger patient cohorts are needed to advance PDFF as an MRI-based biomarker in NMD, with advantages such as its greater dynamic range, enabling the assessment of subtler changes, the amplified objectivity, and the potential of direct correlation to muscle function for selected muscles.

679 Desmoplasia induces T cell exhaustion in cutaneous squamous cell carcinomas

Desmoplastic cutaneous squamous cell carcinomas (cSCC) are a clinically highly aggressive histopathologic variant of cSCC. Desmoplasia, the deposition of abnormal collagen around a tumor, is an independent adverse prognostic factor for disease-specific death in cSCC patients. However, the mechanisms underlying this association with poor outcomes have not been identified in cSCC. Herein we evaluate whether desmoplasia is associated with the suppression of local T cell immune response. Tissue samples from 69 patients who underwent excision for cSCC were graded for desmoplasia by a pathologist using a semi-quantitative grading scale developed in our laboratory.

ASSESSMENT OF CAROTID INTRAPLAQUE HEMORRAGE BY PHOTOACUSTICS IMAGING (PAI) IN PATIENTS UNDERGOING TROMBOENDOARTERIECTOMY: FIRST IN-VIVO HUMAN VALIDATION STUDY

Objective: The aim of the study is the validation of a portable multimodal photoacoustic imaging (PAI) system, for the identification of intraplaque haemorrhage and compare with MRI and histology (gold standard). Design and method: 25 patients with carotid stenosis&gt;70% and clinical indication to tromboendoarterectomy were recruited. Angio-MRI for intraplaque hemorrhage assessment (Cube sequence) was performed. PAI clips (5 seconds, Frame rate 1000/sec) were acquired. Each clip was scored for the presence of PAI signal by means of an integrated scoring system (semiquantitative, from 0 to 12). Semiquantitative grading scales were used to assess plaque histological features of hemorrage and vulnerability. Results: 18 patients had no missing MRI, PAI and histology data and were included in this analysis. Mean age was 73 ± 8 years, 60% men, 80% Caucasians, 92% hypertensives, 60% with a previous stroke. Only 3 plaques out of 21 showed no signs of intraplaque hemorrhage, 4 showed small hemorrage, while 14 (67%) showed large hemorrhages. PAI score (best cut-off &gt;4) correctly classified 14 out of 18 patients (Sensitivity 73.3%, specificity 100%, AUC 0.867). MRI performance was substantially similar (Sensitivity 60%, specificity 100%, AUC 0.800, 12 patients correctly classified), with a non-significant difference in AUC compared to PAI (p = 0.420). Conclusions: In this first in-vivo human study, PAI is able to identify histological intraplaque hemorrhage with an excellent specificity and acceptable sensitivity, equivalent to MRI. The very high specificity, with a low number of false positives, make PAI a good candidate for evaluation of plaques prior to surgery to i.e. reinforce the decision to perform surgery.

Automated morphological phenotyping to characterize knee osteoarthritis progression

Automated morphological phenotyping to characterize knee osteoarthritis progression

Comparison on effects of platelet-rich plasma versus autologous conditioned serum on Achilles tendon healing in a rat model

The aim of this study was to compare the effects of local administrations of platelet-rich plasma (PRP) with autologous conditioned serum (ACS) on Achilles tendon healing in a rat model. In this study, 40 male Sprague-Dawley rats, aged 12 months and weighing 350 to 400 g were used. The rats were divided into three groups (n=10 in each group): a control group and two treatment groups (PRP vs ACS). A standardized procedure was applied for the complete rupture and repair of the Achilles tendon in each group. The PRP group received one dose of PRP on the operative area, and ACS group received ACS at 24, 48, and 72 hours after the surgery. The control group received no injection. Animals were sacrificed 30 days after the operation, and tendon healing in each group was assessed histopathologically based on Bonar's semi-quantitative score and Movin's semi-quantitative grading scale. For the biomechanical analyses, unoperated Achilles tendons of all rats in the control and ACS groups were also harvested, and pulling tests were applied to the specimen to measure the longitudinal axis strength. The highest force value among the data obtained was defined as the maximum strength value (Fmax). The mean Bonar's score was significantly lower in the PRP group (3.8±0.8) than in the ACS (4.8±0.45) and control groups (5.2±0.837) (p=0.0028). The mean Movin's score was significantly lower in the PRP group (7.80±1.49) than in the ACS (9.8±1) and control groups (11.2±2.4) (p=0.029). The ratio of type I collagen was significantly higher in the PRP group (60±6) than in the ACS (52±4.5) and control groups (42±9) (p=0.005). Biomechanical results obtained from operated sites were comparable in terms of Fmax among groups (PRP, 33.93±2.61; ACS, 35.24±3.26; control, 35.69±3.62) (p=0.674). Similarly, the results obtained from unoperated sites were comparable among groups (PRP, 47.71±1.21; ACS, 48.14±2; control, 49.14.69±1.88) (p=0.395). In terms of histopathological results, PRP seems to be more effective than ACS for Achilles tendon healing in rats.

Evaluation of Carotid Plaque Rupture and Neovascularization by Contrast-Enhanced Ultrasound Imaging: an Exploratory Study Based on Histopathology.

A significant portion of ischemic stroke is on account of emboli caused by fibrous cap rupture of vulnerable plaque with intraplaque neovascularization as a significant triggering factor to plaque vulnerability. Contrast-enhanced ultrasound (CEUS) could offer detailed information on plaque surface and intraplaque microvascular. This study aims to comprehensively assess the value of CEUS for the detection of plaque rupture and neovascularization in histologically verified plaques that had been removed from the patients who had undergone carotid endarterectomy (CEA). Fifty-one consecutive subjects (mean age, 67.0 ± 6.5years; 43 [84.3%] men) scheduled for CEA were recruited. Standard ultrasound and CEUS were performed prior to surgery. Based on the direction of the contrast agents that diffuse within the plaques, plaques were divided as "inside-out" direction (contrast agents diffuse from the artery lumen towards the inside of the plaque) and non-inside-out direction. Plaque enhancement was assessed by using a semi-quantitative grading scale (grade 1: no enhancement; grade 2: moderate enhancement; grade 3: extensive enhancement). Plaques were evaluated for histopathologic characteristics according to Oxford Plaque Study (OPS) standard postoperative. Intraplaque neovascularization as manifested by the appearance of CD34-positive microvessels was characterized in terms of microvessel density (MVD), microvessel area (MVA), and microvessel shape (MVS). In 51 plaques, the sensitivity, specificity, positive, and negative predictive values of contrast agent inside-out direction diffusion for the detection of plaque fibrous cap rupture were 87.5%, 92.6%, 91.3%, and 89.3%, respectively. The incidence of cap rupture was significantly higher in contrast agent inside-out direction diffusion than non-inside-out direction diffusion (73.9% vs 25.0%, p< 0.001), and inside-out direction diffusion did exhibit higher frequency of vulnerable plaques (OPS grades 3-4) (95.7% vs 53.6%, p= 0.001). Multivariate logistic regression analysis revealed the contrast agent inside-out direction diffusion as an independent correlate to plaque rupture (OR 8.5, 95% CI 2.4-30.1, p= 0.001). With increasing plaque enhancement, plaque MVD (p< 0.001), plaque MVA (p= 0.012), and percentage of highly irregular-shaped microvessels increased (p< 0.001). Contrast agent inside-out direction diffusion could indicate plaque rupture. The increase in plaque enhancement paralleled increased, larger, and more irregular-shaped microvessels, which may suggest an increased risk of plaque vulnerability.

Semiquantitative Interpretation Criteria for Fluorodeoxyglucose-Positron Emission Tomography/Computed Tomography in Large-Vessel Vasculitis: Pattern Recognition and Correlation with Polymyalgia Rheumatica.

Background:Fluorine-18-fluorodeoxyglucose (18F-FDG)-positron emission tomography/computed tomography (PET/CT) is emerging as a useful imaging modality in suspected large-vessel vasculitis (LVV), owing to its ability to accumulate at the sites of inflammation within the arterial walls. However, there remains scope for standardization of reporting criteria to ensure reproducibility. Recently, a semiquantitative scoring system called “total vascular score” (TVS) has been suggested as a method to standardize and harmonize FDG PET/CT evaluation in LVV patients. The purpose of this study was to assess the clinical utility of the proposed semiquantitative grading scale in LVV patients.Materials and Methods:Patients presenting with clinical symptoms of vasculitis, who had undergone a baseline FDG-PET/CT were evaluated. 18F-FDG uptake in the major vessels was quantified with standardized uptake values (SUVsmax) using four-point scale by three independent nuclear physicians. TVS was calculated based on the calculation of the vascular uptake values with respect to mediastinal blood pool and liver uptake and the number of vessels involved.Results:A total of 106 PET-positive patients (74 males and 32 females) were evaluated. The most frequently involved vessels were thoracic aorta >abdominal aorta >subclavian arteries with mean SUVmax values of 4.05, 3.12, and 2.70, respectively. Mean TVS was 13.18 ± 3.4 (range 03–19) among 276 involved vessels. TVS showed significant positive correlation with erythrocyte sedimentation rate (r = 0.82; P < 0.005). 18 patients showed periarticular FDG uptake, with shoulder joint being the most commonly involved joint.Conclusion:The simplified visual and semiquantitative grading scale for interpretation and reporting classification provides better objectivity in diagnosis, communication with referring clinicians, and planning in patients of LVV.

Safety and effectiveness of cervical vertebroplasty: report of a large cohort and systematic review.

To evaluate retrospectively safety and effectiveness of cervical vertebroplasty (cVP) based on a single-center large cohort. All cVP performed at a single center from January 2001 to October 2014 were included and reviewed. Procedure-related complications (minor and major) were systematically recorded. Effectiveness in terms of analgesia was evaluated using a semi-quantitative grading scale at 1-month follow-up. Risk factors for the occurrence of a procedure-related complication or cement leakage, as well as factors influencing pain relief at 1-month follow-up, were evaluated using a multivariate analysis. One hundred and forty cVP procedures (176 vertebrae) were performed in 130 consecutive patients (88 female, 42 male; mean age = 56 years) during the inclusion period. Among the treated lesions, 80% were bone metastases (mostly from breast cancer), 8% were related to hematological malignancies, and 12% were non-malignant lesions. One fatal complication (0.7%) was related to cement migration in the vertebrobasilar system. Three cervical hematomas were recorded, one of them requiring prolonged oral intubation. The overall rate of major complications was 1.5%. At 1 month, pain reduction was observed in 76% of the cases. Additional surgical fixation was required in 6.1% of the cases. cVP of more than one vertebra during the same session was an independent risk factor for procedure-related complications. Cervical vertebroplasty is a safe technique with an acceptable major complication rate. Its effectiveness in terms of pain relief is good at mid-term follow-up. • Cervical vertebroplasty (cVP) is a safe procedure with a low rate of major complications (1.5%). • cVP provides pain relief in 76% of the cases. • Additional fixation surgery is rarely required after cVP (6.1% of the cases).

Elevated plasma CXCL12/SDF-1 levels are linked with disease severity of postmenopausal osteoporosis.

This study was designed to determine whether plasma CXCL12 levels in postmenopausal osteoporosis (PMOP) patients are related to disease severity. A total of 91 PMOP females were recruited, and 88 postmenopausal non-osteoporotic (PMNOP) women and 90 healthy females were incorporated as controls. Dual-energy X-ray absorptiometry was utilised to explore bone-mineral density (BMD). The Genant semi-quantitative grading scale was used for vertebral fractures, and plasma CXCL12/SDF-1 levels were investigated by ELISA. Plasma TNF-α and C-telopeptide cross-linked collagen type 1 (CTX-1) were also tested. The Oswestry Disability Index (ODI) and a visual analogue scale (VAS) were completed in order to assess clinical severity. Plasma CXCL12 levels were considerably elevated in PMOP females compared to PMNOP women and healthy controls. Plasma CXCL12 concentrations were positively correlated with the Genant grading system. We observed significant and negative correlations of plasma CXCL12 levels with lumbar spine, femoral neck and total hip BMD. Moreover, plasma CXCL12 concentrations were positively correlated to VAS and ODI, as well as plasma TNF-α and CTX-1 levels. In conclusion, elevated plasma CXCL12 levels are correlated with disease severity in PMOP females.

What Are the Effects of Irreversible Electroporation on a Staphylococcus aureus Rabbit Model of Osteomyelitis?

The treatment of osteomyelitis can be challenging because of poor antibiotic penetration into the infected bone and toxicities associated with prolonged antibiotic regimens to control infection. Irreversible electroporation (IRE), a percutaneous image-guided ablation technology in which the targeted delivery of high-voltage electrical pulses permanently damages the cell membrane, has been shown to effectively control bacterial growth in various settings. However, IRE for the management of bone infections has yet to be evaluated. We aimed to evaluate IRE for treating osteomyelitis by assessing (1) the efficacy of IRE to suppress the in vitro growth of a clinical isolate of S. aureus, alone or combined with cefazolin; and (2) the effects of IRE on the in vivo treatment of a rabbit model of osteomyelitis. S. aureus strain UAMS-1 expanded in vitro to the log phase was subjected to an electric field of 2700 V/cm, which was delivered in increasing numbers of pulses. Immediately after electroporation, bacteria were plated on agar plates with or without cefazolin. The number of colony-forming units (CFUs) was scored the following day. ANOVA tests were used to analyze in vitro data. In a rabbit osteomyelitis model, we inoculated the same bacterial strain into the radius of adult male New Zealand White rabbits. Three weeks after inoculation, all animals (n = 32) underwent irrigation and débridement, as well as wound culture of the infected forelimb. Then, they were randomly assigned to one of four treatment groups (n = eight per group): untreated control, cefazolin only, IRE only, or combined IRE + cefazolin. Serial radiography was performed to assess disease progression using a semiquantitative grading scale. Bone and soft-tissue specimens from the infected and contralateral forelimbs were collected at 4 weeks after treatment for bacterial isolation and histologic assessment using a semiquantitative scale. The in vitro growth of S. aureus UAMS-1 was impaired by IRE in a pulse-dependent fashion; the number of CFUs/mL was different among seven pulse levels, namely 0, 10, 30, 60, 90, 120, and 150 pulses. With the number of CFUs/mL observed in untreated controls set as 100%, 10 pulses rendered a median of 50.2% (range 47.1% to 58.2%), 30 pulses rendered a median of 2.7% (range 2.5% to 2.8%), 60 pulses rendered a median of 0.014% (range 0.012% to 0.015%), 90 pulses rendered a median of 0.004% (range 0.002% to 0.004%), 120 pulses rendered a median of 0.001% (range 0.001% to 0.001%), and 150 pulses rendered a median of 0.001% (range 0.000% to 0.001%) (Kruskal-Wallis test: p = 0.003). There was an interaction between the effect of the number of pulses and the concentration of cefazolin (two-way ANOVA: F [8, 30] = 17.24; p < 0.001), indicating that combining IRE with cefazolin is more effective than either treatment alone at suppressing the growth of S. aureus UAMS-1. Likewise, the clinical response in the rabbit model (the percentage of animals without detectable residual bacteria in the bone and surrounding soft tissue after treatment) was better in the combination group than in the other groups: control, 12.5% (one of eight animals); IRE only, 12.5% (one of eight animals); cefazolin only, 25% (two of eight animals); and IRE + cefazolin, 75% (six of eight animals) (two-sided Fisher's exact test: p = 0.030). IRE effectively suppressed the growth of S. aureus UAMS-1 and enhanced the antibacterial effect of cefazolin in in vitro studies. When translated to a rabbit osteomyelitis model, the addition of IRE to conventional parenteral antibiotic treatment produced the strongest response, which supports the in vitro findings. Our results show that IRE may improve the results of standard parenteral antibiotic treatment, thus setting the stage for models with larger animals and perhaps trials in humans for validation.

Evaluation of Percutaneous Intradiscal Amniotic Suspension Allograft in a Rabbit Model of Intervertebral Disc Degeneration.

A laboratory study using a rabbit annular puncture model of intervertebral disc degeneration (IDD). The aims of this study were to assess whether an amniotic suspension allograft (ASA) containing particulated human amnion and amniotic fluid derived cells regains intervertebral disc height and morphology and improves histologic scoring in a rabbit model of IDD. In contrast to current surgical interventions for IDD, in which the primary goal is to relieve symptomatic pain, one novel strategy involves the direct injection of anabolic cytokines. Current therapies for IDD are limited by both the short half-life of therapeutic proteins and general decline in anabolic cell populations. Intervertebral discs in New Zealand white rabbits were punctured using 18-gauge needle under fluoroscopic guidance. Four weeks post-puncture, two groups of rabbits were injected with either ASA or a vehicle/sham control, while a third group was untreated. Weekly radiographs were obtained for 12 weeks to assess disc height index (DHI). Magnetic resonance imaging (MRI) T2 relaxation time was evaluated at weeks 4 and 12 to assess morphological changes. Histologic sections were evaluated on a semi-quantitative grading scale. Before treatment at week 4, DHIs and normalized T2 relaxation times between the three groups were not significantly different. At week 12, ASA-treated rabbits exhibited significantly greater DHIs and MRI T2 relaxation times than vehicle and untreated control groups. The ASA group had higher mean histologic score than the vehicle group, which demonstrated extensive fiber disorganization and delamination with reduced proteoglycan staining on histology. Minimally invasive intervention with intradiscal injection of ASA was successful in reducing IDD in a reproducible rabbit model, with significant improvement in disc height and morphology when compared with vehicle and untreated control groups on radiographic and MRI analyses. N/A.

Digital Quantification of Epidermal Protein Expression in Paraffin-Embedded Tissue Using Immunohistochemistry.

Current methods of assessing immunohistochemistry center on semiquantitative visual grading scales. More objective methods utilizing digital quantification offer superior precision and, presumably, higher confidence with image comparison. However, their cost often remains prohibitive, and there is little customizability to separate subsections of interest in the tissue. Here we describe a method using two open-source software programs to analyze the intensity and density of signals in immunohistochemistry-stained tissue sections that account for tissue heterogeneity and allow for direct comparison between two samples. This method allows for quantitative assessment of epidermal protein expression. We herein demonstrate this workflow using an epidermal stain tothymic stromal lymphopoietin.