Serum Aldolase Research Articles

Real world data of cabozantinib in patients with hepatocellular carcinoma: Focusing on dose setting and modification.

To investigate the outcomes of cabozantinib in patients with unresectable hepatocellular carcinoma (uHCC), focusing on dose setting and modification. We retrospectively analyzed 34 Japanese patients who received cabozantinib for uHCC. Trough concentrations (Ctrough) of cabozantinib were also measured weekly for 6 weeks in the 18 patients. Sixteen patients received ≥40 mg (high-dose group), and 18 patients received 20 mg (low-dose group). Dose escalations were performed in 27.8% of the patients in the low-dose group. Although median duration of the first dose reduction or interruption in the low-dose group was twice that in the high-dose group (28 vs. 14 days, p < 0.001), there were no significant differences in the relative dose intensity (RDI) during 6 weeks, progression free survival (PFS), and overall survival (p = 0.162, p = 0.950, p = 0.817, respectively) between the two groups. Patients who received RDI during 6 weeks ≥33.4% showed a trend toward longer median PFS (p = 0.054). Each serum aldolase value during the 6 weeks was significantly correlated with the Ctrough at any point (r = 0.500, p < 0.001). In multivariate analyses, aldolase ≥8.7 U/L within 2 weeks was significantly associated with the very early dose reduction or interruption (odds ratio 20.0, p = 0.002). An initial dose of 20 mg cabozantinib could be a safe option in Japanese patients. The serum aldolase value could be useful for making appropriate dose modifications of cabozantinib.

Seronegative Polyarticular Juvenile Idiopathic Arthritis: A Case Report.

Juvenile idiopathic arthritis (JIA) is a chronic rheumatologic childhood disorder of unknown etiology usually presenting with peripheral arthritis. Usually involving the small joints symmetrically, polyarticular JIA is characterized by involvement of 5 or more peripheral joints and is further categorized depending on RA Factor positivity. We aim to report a case of seronegative polyarticular JIA to increase awareness regarding this disease. A 15-year-old boy presented with 3-month history of bilateral symmetrical joint pains, swelling and morning stiffness lasting 3-4 hours. For his ankle and knee pain, he had consulted a local doctor who suspected Rheumatic Fever and started penicillin and analgesics resulting in minimal improvement. He was a student but was unable to attend school in last 2 months due to this illness and had become bed-bound for last 2 weeks. On examination, there was swelling and tenderness of ankles, knees, elbows, wrists, metacarpophalangeal and proximal interphalangeal joints bilaterally. On investigation, ESR, CRP and serum ferritin were raised with normal complete blood count, urinalysis, liver and renal function tests, ASO test, blood and urine cultures, TSH, serum CPK and serum aldolase. Echocardiography did not reveal any abnormalities. His ANA, RA factor and Anti-CCP antibodies were negative. He was diagnosed with Seronegative Polyarticular Juvenile Idiopathic Arthritis and started on oral prednisolone and methotrexate. Currently the patient is asymptomatic on methotrexate 20mg/week. Our patient with symmetrical polyarthritis had negative ANA, RA Factor and Anti-CCP antibodies leading to a diagnosis of seronegative polyarticular JIA. Early screening and prompt management of JIA is required to improve prognosis and reduce risk of disability. The differential diagnosis of childhood joint pains is wide and variable including diseases of autoimmune, cardiac and haematological systems. For diagnosis of JIA, arthritis persisting for more than 6 weeks with onset of symptoms prior to 16 years of age is required with exclusion of secondary causes of joint inflammation. Keywords: Juvenile Idiopathic Arthritis, ANA, RA Factor, Anti-CCP antibodies, Methotrexate.

Colchicine-Induced Neuromyopathy with Myotonic Discharges in a Patient Using Concomitant Diuretics

Colchicine is a widely used anti-inflammatory medication, but its neuromuscular adverse effects are under-recognized. One month ago, a 70-year-old woman presented to our clinic for an evaluation of general weakness. She had been taking colchicine and diuretics daily. The weakness, which began in both thighs and lower legs approximately 4 weeks before her visit, had progressively worsened. The patient also experienced mild paresthesia and hypoesthesia in both arms and legs. Her serum creatine kinase and aldolase levels were elevated, and nerve conduction studies indicated a motor-dominant sensorimotor polyneuropathy of the axonal type. Needle electromyography showed prominent fibrillation potentials, positive sharp waves, and myotonic discharges. Suspecting colchicine-induced neuromyopathy, we discontinued the colchicine and diuretics, after which her symptoms resolved.

Poor prognostic factors for relapse of interstitial lung disease with anti-aminoacyl-tRNA synthetase antibodies after combination therapy.

This study aimed to identify useful clinical indicators for predicting the relapse of interstitial lung disease (ILD) complicated with anti-aminoacyl-tRNA synthetase (ARS) antibodies (anti-ARS-ILD), being treated with prednisolone and calcineurin inhibitors. Fifty patients with anti-ARS-ILD were enrolled between October 2014 and August 2022. All patients were treated with prednisolone and calcineurin inhibitors as remission induction therapy and followed up for over a year with these combination therapies. We examined patients who experienced ILD relapse after immunosuppressive treatment. We explored the risk factors for predicting ILD relapse in these patients by comparing demographic, clinical, laboratory, and radiological findings and treatments between the relapsed and non-relapsed groups on admission. Of the 50 patients, 19 (38%) relapsed during a median follow-up of 4.8 years. Univariate and multivariate Cox regression analyses identified the presence of acute/subacute (A/S)-ILD, higher serum aldolase (ALD) and surfactant protein-D (SP-D) levels, and lower %forced vital capacity (FVC) as risk factors for relapse in patients with anti-ARS-ILD. Using the receiver operating curve analysis, ALD ≥6.3 U/L, SP-D ≥207 ng/mL, and %FVC ≤76.8% were determined as the cut-off levels for indicating a poor prognosis. The 5-year relapse rate was significantly higher in patients with A/S-ILD, serum ALD≥6.3 U/L, serum SP-D ≥207 ng/mL, or %FVC of ≤76.8% than in those without these parameters. (P=0.009, 0.0005, 0.0007, 0.0004, respectively) Serum ALD levels were significantly correlated with the disease activity indicators of anti-ARS-ILD. The presence of A/S-ILD, higher serum ALD and SP-D levels, and lower %FVC are useful indicators for predicting anti-ARS-ILD relapse.

Detection of proteins with ascorbic acid-capped gold nanoparticles: a simple and highly sensitive colorimetric assay.

We report a simple and highly sensitive colorimetric method for the detection and quantification of proteins, based on the aggregation of ascorbic acid (AA) capped gold nanoparticles (AuNPs) by proteins. The interactions between our AuNPs and nine different proteins of various sizes and shapes (cytochrome C (12 kDa), lysozyme (14.3 kDa), myoglobin (17 kDa), human serum albumin (66 kDa), bovine serum albumin (66.4 kDa), human transferrin (80 kDa), aldolase (160 kDa), catalase (240 kDa), and human H-ferritin (500 kDa)) generated similar AuNPs-protein absorption spectra in a concentration-dependent manner in the range of 1-15 nM. Upon the addition of a protein, the UV-visible spectra of AuNPs-protein conjugates shifted from 524 nm for the AuNps alone to longer wavelength (600-750 nm) due to the presence of one of these proteins. This bathochromic shift is accompanied by a color change from a cherry red, to dark purple, and then light grey or colorless if excess protein has been added, indicating the formation of AuNPs-protein conjugates followed by protein-induced aggregation of the AuNPs. High-resolution transmission electron microscopy images revealed uniformly distributed spherical nanoparticles with an average size of 27.5 ± 15.2 nm, increasing in size to 39.6 ± 12.9 nm upon the addition of a protein, indicating the formation of AuNPs-protein conjugates in solution. A general mechanism for the protein-induced aggregation of our AuNPs is proposed. The consistent behavior observed with the nine proteins tested in our study suggests that our assay can be universally applied for the quantification of pure proteins in a solution, regardless of size, shape, or molecular weight.

Disease spectrum of myopathies with elevated aldolase and normal creatine kinase.

Elevation of serum creatine kinase (CK) or hyperCKemia is considered a biological marker of myopathies. However, selective elevation of serum aldolase with normal CK has been reported in a few myopathies, including dermatomyositis, immune-mediated myopathy with perimysial pathology and fasciitis with associated myopathy. The aim was to investigate the disease spectrum of myopathies with isolated aldolase elevation. Medical records were reviewed to identify patients >18 years old seen between December 1994 and June 2020 who had pathologically proven myopathies with elevated aldolase and normal CK level. Patients with alternative causes of aldolase elevation were excluded. Thirty-four patients with various types of myopathies were identified. Myopathies were treatable in 27 patients. The three most common etiologies were dermatomyositis (n = 8), overlap myositis (n = 4) and nonspecific myopathy (n = 4). Perimysial pathology comprising inflammation, fragmentation, vasculitis, calcified perimysial vessels or extracellular amyloid deposition was found in 17/34 patients (50%). Eight dermatomyositis patients with selective elevated aldolase were compared to 24 sex- and age-matched patients with dermatomyositis and hyperCKemia. Dermatomyositis patients with normal CK significantly (p < 0.05) had less frequent cutaneous involvement (50.0% vs. 100.0%) and fibrillation potentials (50.0% vs. 90.5%) but higher median erythrocyte sedimentation rate (33.5 vs. 13.5 mm/h) and more common perifascicular mitochondrial pathology (37.5% vs. 4.2%). Isolated aldolase elevation can be found in a greater variety of myopathies than initially thought and most were treatable. Dermatomyositis is the most common myopathy with selective elevation of aldolase in our cohort, which features some unique characteristics compared to dermatomyositis with hyperCKemia.

Association of Juvenile Dermatomyositis Disease Activity With the Expansion of Blood Memory B and T Cell Subsets Lacking Follicular Markers.

This study was undertaken to identify blood markers of juvenile dermatomyositis (DM) disease activity (DA), which are needed to improve disease management. The study comprised a total of 123 juvenile DM patients and 53 healthy controls. Results of laboratory tests (aldolase, creatinine kinase, lactate dehydrogenase [LDH], aspartate aminotransferase) and clinical measures of DA in patients with juvenile DM, including the Manual Muscle Testing in 8 muscles (MMT-8), Childhood Myositis Assessment Scale (CMAS), and disease activity scores (DAS) (total DAS for juvenile DM, the muscle DAS, and the skin DAS), were recorded when available. Surface phenotype of peripheral blood mononuclear cells was assessed using flow cytometry. Whole blood transcriptional profiles were studied using either RNA-sequencing or microarrays. Differential gene expression was determined using DESeq and compared by pathway and gene ontology analyses. Conventional memory (CD27+IgD-) B cells expressing low CXCR5 levels (CXCR5low/- CM B cells) were significantly increased in frequency and absolute numbers in 2 independent cohorts of juvenile DM patients compared with healthy controls. The frequency of CD4+ Th2 memory cells (CD45RA-CXCR5-CCR6-CXCR3-) was also increased in juvenile DM, especially in patients who were within <1 year from diagnosis. The frequency of CXCR5low/- CM B cells correlated with serum aldolase levels and with a blood interferon-stimulated gene transcriptional signature. Furthermore, both the frequency and absolute numbers of CXCR5low/- CM B cells correlated with clinical and laboratory measures of muscle DA (MMT-8, CMAS, aldolase, and LDH). These findings suggest that both CM B cells lacking the CXCR5 follicular marker and CXCR5- Th2 cells represent potential biomarkers of DA in juvenile DM and may contribute to its pathogenesis.

Immunopathological features of myopathy associated with small-to-medium-sized vessel vasculitis and differences from autoimmune myositis.

Patients with systemic vasculitis may develop myalgia as an initial symptom. However, the immunopathology of vasculitic myopathy remains unclear. We investigated the immunopathological features of skeletal muscle in small-to-medium-sized vessel vasculitis. We analysed muscle tissue biopsies from 15 patients with vasculitis, including antineutrophil cytoplasmic antibodyassociated vasculitis and polyarteritis nodosa, and 15 patients with autoimmune myositis (AIM), including polymyositis and immune-mediated necrotising myopathy, as comparison disease controls. Immunohistochemical staining for CD56/neural cell adhesion molecule (NCAM), major histocompatibility complex class I, C5b-9/membrane attack complex (MAC), and CD31 was performed. The vascularity score was defined as the total number of CD31-expressing blood vessels. The association between CD56/NCAM-expressing myofibers and clinical findings was evaluated in patients with vasculitis. Patients with vasculitis had a significantly lower frequency of CD56/NCAM-expressing myofibers than those with AIM and a positive correlation between the frequency of CD56/NCAM-expressing myofibers and serum aldolase levels. Patients with vasculitis had significantly fewer major histocompatibility complex class I-expressing myofibers and C5b-9/MAC deposits on the sarcolemma than those with AIM. C5b-9/MAC deposits in blood vessels were observed in >70% of patients with vasculitis. Patients with vasculitis had significantly higher vascularity scores in the endomysium than those with AIM. Patients with vasculitis demonstrated mild myofiber damage based on the lower involvement of CD56/NCAM-expressing myofibers compared to those with AIM. Complement component deposits on the vessel walls and hypervascularity in the endomysium areas may be immunopathological features of vasculitic myopathy.

Impact of Cabozantinib Exposure on Proteinuria and Muscle Toxicity in Patients with Unresectable Hepatocellular Carcinoma.

This prospective study investigated the impact of cabozantinib exposure on proteinuria and muscle toxicity, in a cohort of 14 Japanese patients with unresectable hepatocellular carcinoma (uHCC). We measured the trough concentration of cabozantinib (Ctrough) weekly for 6 weeks after starting treatment. Although the initial dose was less than 60 mg in most cases, dose interruption occurred in 79%, primarily because of proteinuria and/or malaise. The median and coefficient of variation of maximum Ctrough at 7-42 d were 929.0 ng/mL and 59.2%, respectively. The urinary protein-to-creatinine ratio (UPCR), serum creatine kinase, and serum aldolase values were all significantly elevated following treatment. Moreover, maximum changes in serum creatine kinase and aldolase were significantly associated with maximum Ctrough (r = 0.736, p &lt; 0.01; r = 0.798, p &lt; 0.001; respectively). Receiver operating characteristic (ROC) curve analysis showed that changes in serum creatine kinase ≥70.5 U/L and aldolase ≥6.1 U/L from baseline relatively accurately predicted inclusion in the high-maximum Ctrough (≥929.0 ng/mL) group, with an area under the ROC of 0.929 and 0.833, respectively. Measurement of serum creatine kinase and aldolase may increase the clinical usefulness of cabozantinib treatment for uHCC and help alleviate difficulties with dose adjustments.

Adherence to Choosing Wisely Campaign Guidelines at Three Health Systems

Abstract Introduction/Objective With rising healthcare costs in the United States, there has been a push for lab stewardship to improve the quality of patient care while reducing costs. To optimize the use of clinical laboratory testing, the ASCP working with other medical specialty organizations, developed the Choosing Wisely Campaign to promulgate evidence-based guidelines to optimize clinical laboratory testing. Methods/Case Report We examined adherence to three Choosing Wisely guidelines over a four-year period (2017- 2020), through queries of internal cost accounting databases to return aggregate volumes as well as variable and total costs at three large academic health systems. We measured concurrent orders for: 1) erythrocyte sedimentation rate (ESR) with C-reactive protein (CRP), 2) serum/plasma amylase with lipase, and 3) free thyroxine (FT4) and/or total triiodothyronine (TT3) with thyroid stimulating hormone (TSH) when the TSH is within the reference range (using an frequency estimate of 85% based on other studies). We also examined another guideline for concurrent orders for serum aldolase with creatine kinase (CK). We also quantified aggregate variable costs for the non-recommended test in each Choosing Wisely guideline (amylase, ESR, FT4 and/or TT3), and for serum aldolase when ordered with CK. Results (if a Case Study enter NA) Over the four-year period, there were 322,853 unnecessary tests based on these four guidelines (120,587 ESR and CRP, 30,444 amylase and lipase, 164,818 FT4 and/or TT3 with TSH, and 7,004 aldolase). Overall, unnecessary testing decreased between 2017 and 2020 for amylase with lipase, remained essentially unchanged for aldolase, and increased for the other two test guideline scenarios. The largest changes were concurrent orders for amylase and lipase at one health system (38% decrease), and orders for TT3 with a normal TSH result at another health system (324% increase). The four-year variable cost of these unnecessary tests was $1,215,309 ($303,827 mean annual cost), resulting in potential annual variable cost savings of $101,276 for each health system for the four guidelines we examined. Variable costs for unnecessary testing increased by 16.5% ($45,571) over the four-year period. Conclusion Guideline-based unnecessary testing remains as a target to improve laboratory diagnostic testing. There is potential to realize significant achievable cost savings if guidelines are implemented and maintained.

IDIOPATHIC INTERSTITIAL PNEUMONITIS: A MANIFESTATION OF SYSTEMIC SCLEROSIS SINE SCLERODERMA

TOPIC: Pulmonary Manifestations of Systemic Disease TYPE: Fellow Case Reports INTRODUCTION: Systemic Sclerosis (SSc) is a multisystem disease characterized by vascular dysfunction and progressive fibrosis. Systemic sclerosis sine scleroderma (ssSSc) is a rare subtype of SSc which lacks the hallmark feature of the disease i.e., scleroderma. We present a case of systemic sclerosis sine scleroderma manifesting as interstitial pneumonitis. CASE PRESENTATION: A 76-year-old Caucasian female presented to the clinic for evaluation of dyspnea and decreased exercise tolerance since recent TAVR performed 1 month ago. She had a history of Raynaud phenomenon, renal artery stenosis, CKD 4, and HTN. She was hypoxic at rest with SpO2 87% on room air, the rest of the vitals were normal. Chest x-ray showed coarse reticular opacities. Pulmonary function tests (PFTs) showed mild restriction (FEV1/FVC 85%, TLC 74% of predicted) and severely decreased diffusing capacity for carbon monoxide (DLCO) 31% of predicted (N: 80-120%). High-resolution CT chest demonstrated bilateral middle and lower lung zone reticular opacities, and traction bronchiectasis. Pre TAVR right heart catheterization was consistent with pulmonary arterial hypertension (PAH). Her PAH was not considered as the cause of all her symptoms.Further workup showed positive serum ANA 1:1280 (N: <1:80), and weakly positive RNA polymerase III antibody at 30.2U (N: < 20U). Anti CCP, anti- dsDNA, anticentromere antibodies, rheumatoid factor, serum aldolase, CRP, complement levels, ESR, and ANCA panel were normal. Although she did not have any skin findings suggestive of scleroderma, based on serological and imaging abnormalities she was diagnosed with ssSSc presenting as interstitial pneumonitis. She was treated initially with prednisone followed by mycophenolate for 5 months. Despite stabilization of PFTs, her respiratory status continues to decline from worsening PAH. She remains on supplemental oxygen, sildenafil, and treprostinil. DISCUSSION: ssSSc is a subset of SSc affecting 5-10% of patients, with clinical and laboratory features resembling limited SSc (lcSSc) subset. Lungs are the second most common organ involved. ILD followed by PH are the most common presentations. Patients with ssSSc are reported to have a greater degree of dyspnea, and reduced DLCO (<70% of predicted normal). Other predominant features include a positive ANA, restrictive lung disease, and Raynaud's phenomenon, which were present in our patient. CONCLUSIONS: Systemic sclerosis should be considered in patients with idiopathic ILD even in the absence of skin involvement. REFERENCE #1: Poormoghim H, Lucas M, Fertig N, Medsger TA Jr. Systemic sclerosis sine scleroderma: demographic, clinical, and serologic features, and survival in forty-eight patients. Arthritis Rheum. 2000 Feb;43(2):444-51. doi: 10.1002/1529-0131(200002)43:2<444: AID-ANR27>3.0.CO;2-G. PMID: 10693887. REFERENCE #2: Diab S, Dostrovsky N, Hudson M, Tatibouet S, Fritzler MJ, Baron M, Khalidi N; Canadian Scleroderma Research Group. Systemic sclerosis sine scleroderma: a multicenter study of 1417 subjects. J Rheumatol. 2014 Nov;41(11):2179-85. doi: 10.3899/jrheum.140236. Epub 2014 Oct 1. PMID: 25274885. REFERENCE #3: Kucharz EJ, Kopeć-Mędrek M. Systemic sclerosis sine scleroderma. Adv Clin Exp Med. 2017 Aug;26(5):875-880. doi: 10.17219/acem/64334. PMID: 29068586. DISCLOSURES: No relevant relationships by Mirza Ali, source=Web Response No relevant relationships by Shujaa Faryad, source=Web Response No relevant relationships by Hasan Shoaib, source=Web Response

Anti-MDA5 autoantibodies associated with juvenile dermatomyositis constitute a distinct phenotype in North America.

Myositis-specific autoantibodies have defined distinct phenotypes of patients with juvenile myositis (JIIM). We assessed the frequency and clinical significance of anti-melanoma differentiation-associated gene 5 (MDA5) autoantibody-associated JIIM in a North American registry. Retrospective examination of the characteristics of 35 JIIM patients with anti-MDA5 autoantibodies was performed, and differences from other myositis-specific autoantibody groups were evaluated. Anti-MDA5 autoantibodies were present in 35/453 (7.7%) of JIIM patients and associated with older age at diagnosis, and lower serum creatine kinase and aldolase levels. Patients with anti-MDA5 autoantibodies had more frequent weight loss, adenopathy, arthritis, interstitial lung disease (ILD), and less frequent falling compared with anti-transcriptional intermediary factor 1 (TIF1), anti-nuclear matrix protein 2 (NXP2)and myositis-specific autoantibody/myositis-associated autoantibody-negative patients. They had a different season of diagnosis and less frequent mechanic's hands and ILD compared with those with anti-synthetase autoantibodies. Anti-MDA5 patients received fewer medications compared with anti-TIF1, and corticosteroid treatment was shorter compared with anti-TIF1 and anti-nuclear matrix protein 2 autoantibody groups. The frequency of remission was higher in anti-MDA5 than anti-synthetase autoantibody-positive JIIM. In multivariable analyses, weight loss, arthritis and arthralgia were most strongly associated with anti-MDA5 autoantibody-positive JIIM. Anti-MDA5 JIIM is a distinct subset, with frequent arthritis, weight loss, adenopathy and less severe myositis, and is also associated with ILD. Anti-MDA5 is distinguished from anti-synthetase autoantibody-positive JIIM by less frequent ILD, lower creatine kinase levels and differing seasons of diagnosis. Anti-MDA5 has comparable outcomes, but with the ability to discontinue steroids more rapidly and less frequent flares compared with anti-TIF1 autoantibodies, and more frequent remission compared with anti-synthetase JIIM patients.

Assessment of Optimal Screening Tests for the Detection of an Inflammatory Myositis-associated Interstitial Lung Disease.

BackgroundInterstitial lung disease (ILD) is a common pathologic consequence of the idiopathic inflammatory myopathies, and it may be the initial presentation of autoimmune disease in many cases. There are no well-established guidelines to direct the evaluation of this disease in these cases. This study looked at the utility of four common serologic tests to screen for a myositis-associated ILD.MethodsThis is a single institution retrospective analysis of four common serologic tests (antinuclear antibody [ANA], creatine kinase [CK], aldolase, and anti-Sjögren's syndrome A [anti-SSA]) to detect a positive antibody on an extended myositis antibody panel.ResultsThe serum aldolase was the most sensitive test to detect the presence of a positive antibody on an extended myositis antibody panel with a sensitivity of 54.5%. The anti-SSA was the least sensitive at 21.4%. A positive result for anti-SSA antibodies was associated with a 100% positive predictive value when all other screening tests (ANA, aldolase, and CK) were also positive. ConclusionNo single screening test was sufficient for the evaluation of a myositis-associated ILD. A positive serum aldolase had higher sensitivity, and a positive SSA had a high positive predictive value when other screening markers were also elevated, but clinicians still need to maintain a high index of suspicion for myositis-associated ILD.

Chronic Graft Versus Host Myopathies: Noninflammatory, Multi-Tissue Pathology With Glycosylation Disorders.

Myopathies during chronic graft-versus-host disease (cGvHD) aresyndromes for which tissue targets and mechanisms of muscle damage remain incompletely defined. This study reviewed, and pathologically analyzed, 14 cGvHD myopathies, comparing myopathology to other immune myopathies. Clinical features in cGvHD myopathy included symmetric, proximal weakness, associated skin, gastrointestinal and lung disorders, a high serum aldolase (77%), and a 38% 2-year survival. Muscle showed noninflammatory pathology involving all 3 tissue components. Perimysial connective tissue had damaged structure and histiocytic cells. Vessel pathology included capillary loss, and reduced α-l-fucosyl and chondroitin sulfate moieties on endothelial cells. Muscle fibers often had surface pathology. Posttranslational glycosylation moieties on α-dystroglycan had reduced staining and abnormal distribution in 86%. Chondroitin-SO4 was reduced in 50%, a subgroup with 3-fold longer times from transplant to myopathy, and more distal weakness. cGvHD myopathies have noninflammatory pathology involving all 3 tissue components in muscle, connective tissue, small vessels, and myofibers. Abnormal cell surface glycosylation moieties are common in cGvHD myopathies, distinguishing them from other immune myopathies. This is the first report of molecular classes that may be immune targets in cGvHD. Disordered cell surface glycosylation moieties could produce disease-related tissue and cell damage, and be biomarkers for cGvHD features and activity.

Clinical and immunological analyses of immune checkpoint inhibitor-associated neuromuscular (NM) toxicities.

e14118 Background: A rare and often fatal spectrum of NM immune-related adverse events (irAEs) includes myositis, myocarditis, and myasthenia gravis (MG). A deeper understanding of their clinical and immunologic features may improve outcomes. Methods: Sixteen patients (pts) with genitourinary cancers who were treated with ICI and developed myositis, myocarditis, MG, or overlap syndrome (≥2) were identified. Diagnostic criteria included serum CK (n=16), aldolase (n=14), troponin (n=12), BNP (n=15), autoantibodies (n=16), and cytokines (n=9). Biopsies of skeletal muscle (n=13) and endomyocardium (n=4) were also obtained. Immune profiling is being performed on blood and tissue for pts consented to an institutional-approved laboratory protocol. Results: ICI-associated NM irAEs consisted of isolated myositis (n=8, 50%), myocarditis (n=1, 6%), MG (n=1, 6%), or overlap syndrome (n=6, 38%), with the most common presenting symptom being weakness (n=9, 56%). CK levels were elevated in 15 (94%) pts, and autoantibodies were detected in 9 (56%). Muscle biopsies showed predominantly T cell infiltration. All 16 pts received corticosteroids, 12 (75%) were treated with other immunosuppressive agents, and 14 (88%) underwent plasmapheresis. Five pts (50%) with isolated irAEs achieved symptom resolution, and none in the overlap group. (Table) Conclusions: NM irAEs occurred more frequently with combination ICI, and were associated with weakness, and elevated CK level and serum autoantibodies. Both T and B cells appear to drive disease pathogenesis. The presence of one toxicity on the spectrum warrants evaluation for associated irAEs. Compared to isolated NM irAEs, the overlap syndrome appears to be more treatment refractory. Current guidelines require refinement to identify effective treatments to improve outcomes. [Table: see text]

Aldolase - From Biochemistry to Laboratory Medicine

Aldolase (ALD) (fructose-1,6-bisphosphate aldolase ) is a 160 kDa, enzyme which catalyzes the conversion of fructose 1-6-biphosphate in glyceraldehyde 3-phosphate and dihydroxyacetone phosphate in the glycolytic metabolic pathway. There are also experimental data suggesting that nuclear ALD isoenzyme A might play an important role in cell proliferation. At the present time, the most useful serum markers of muscle injury following intense, prolonged exercise are: creatine kinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase myoglobin and troponin. Although serum ALD is not usually measured yet, it may be used together with CK to evaluate the status of muscle adaptation to training. Recent studies offered ALD a new perspective, as a future valuable biomarker in monitoring the evolution of muscle crush injuries, in order to prevent silent, but progressiv muscle fibers necrosis after injury. It has also been shown that ALD was an independent clinical prognostic marker in many other human cancers, being involved in some well-known signaling pathways.

Small Extracellular Vesicles in Rat Serum Contain Astrocyte-Derived Protein Biomarkers of Repetitive Stress.

BackgroundStress precipitates mood disorders, characterized by a range of symptoms present in different combinations, suggesting the existence of disease subtypes. Using an animal model, we previously described that repetitive stress via restraint or immobilization induced depressive-like behaviors in rats that were differentially reverted by a serotonin- or noradrenaline-based antidepressant drug, indicating that different neurobiological mechanisms may be involved. The forebrain astrocyte protein aldolase C, contained in small extracellular vesicles, was identified as a potential biomarker in the cerebrospinal fluid; however, its specific origin remains unknown. Here, we propose to investigate whether serum small extracellular vesicles contain a stress-specific protein cargo and whether serum aldolase C has a brain origin.MethodsWe isolated and characterized serum small extracellular vesicles from rats exposed to restraint, immobilization, or no stress, and their proteomes were identified by mass spectrometry. Data available via ProteomeXchange with identifier PXD009085 were validated, in part, by western blot. In utero electroporation was performed to study the direct transfer of recombinant aldolase C-GFP from brain cells to blood small extracellular vesicles.ResultsA differential proteome was identified among the experimental groups, including aldolase C, astrocytic glial fibrillary acidic protein, synaptophysin, and reelin. Additionally, we observed that, when expressed in the brain, aldolase C tagged with green fluorescent protein could be recovered in serum small extracellular vesicles.ConclusionThe protein cargo of serum small extracellular vesicles constitutes a valuable source of biomarkers of stress-induced diseases, including those characterized by depressive-like behaviors. Brain-to-periphery signaling mediated by a differential molecular cargo of small extracellular vesicles is a novel and challenging mechanism by which the brain might communicate health and disease states to the rest of the body.

Clinical significance of subcutaneous fat and fascial involvement in juvenile dermatomyositis

Objectives: Subcutaneous involvement, including calcinosis and panniculitis, is a more common complication in juvenile dermatomyositis (JDM) than in adult dermatomyositis. Magnetic resonance imaging (MRI) is useful for evaluating disease distribution. We investigated the clinical significance of subcutaneous involvement in JDM.Methods: Thighs and hips in 18 newly diagnosed JDM patients were evaluated with fat-suppression MRI. Bilateral muscle, fascial and subcutaneous fat involvement were scored from 0 to 8 points according to the severity of distribution on MRI. Associations between clinical manifestations, serum muscle enzymes, and MRI scores were also evaluated.Results: Abnormal MRI findings in muscle, fascia and subcutaneous fat were observed in 18, 18, and 10 patients, respectively. Subcutaneous fat scores were significantly higher in early-diagnosed JDM patients (diagnosed less than 2 months from onset) than in late-diagnosed JDM patients (diagnosed later) (p = .025). Serum aldolase was elevated in all patients, although only eight demonstrated elevated serum creatine phosphokinase. Serum aldolase was significantly correlated with MRI scores for subcutaneous fat (p < .0001, ρ = .787) and fascia (p = .013 ρ = 0.574), but not muscle. Additionally, serum aldolase was significantly correlated with serum triglycerides (p = .009, ρ = 0.629).Conclusion: Subcutaneous fat involvement is a characteristic finding in early-diagnosed JDM and correlates with elevated serum aldolase.

Voltammetric and chronopotentiometric protein structure-sensitive analysis

Previously we showed that constant current chronopotentiometric stripping (CPS) is convenient for protein analysis based on the ability of some amino acid residues to catalyze hydrogen evolution on mercury-containing electrodes. This method showed a remarkable sensitivity to changes in protein structures, including protein denaturation and even small protein damage. Here we used normal pulse voltammetric stripping (NPVS) with bare and dithiothreitol-modified hanging mercury drop electrode. We found that NPV pulses denatured the surface-attached protein but we showed conditions under which this method was able to distinguish native and denatured proteins with sensitivity approaching that of CPS. Using NPVS it was possible to follow bovine serum albumin (BSA) thermal denaturation as well as to investigate the effect of NPV pulses on the structure of the surface-attached protein. In addition to BSA we studied several proteins, such as human serum albumin, ovalbumin, urease, aldolase, concanavalin A, histone and vasopressin peptide. Our results suggest that CPS remains the method-of-choice for studies of changes in protein structure and of biochemical processes, such as DNA-protein specific binding, lectin-glycoprotein interactions, detection of protein damage, etc., while voltammetric methods, such as NPVS may suit better for investigation of the processes which proteins undergo at the electrode surface.

MUSCULAR DYSTROPHIES

Objectives: Muscular dystrophies are classified into different types based ontheir age of onset, clinical severity, rate of progression, distribution of muscles weakness,pattern of inheritance and the genes involved. As muscular dystrophies are relativelyuncommon disorders, very little work has been done in Pakistan. This study has classified (forthe first time in Pakistan) the patients with different types of muscular dystrophies by clinicaland biochemical correlation. Study Design: Observational, descriptive study. Setting:Departments of Morbid Anatomy and Histopathology and Human Genetics and MolecularBiology at University of Health Sciences Lahore. Period: Three years’ (March, 2012-March2015). Methodology: A total of 100 patients comprising of all age groups and both gendersand with strong clinical suspicion of muscular dystrophy, with or without positive family history,were included. Detailed clinical history and physical examination findings were recordedfollowed by estimation of serum Creatine Phosphokinase (CPK), Lactate Dehyrogenase(LDH) and Aldolase levels. Results: A total of 77 males (M) and 23 females (F) were included.Mean age of the patients was 12.1990 + 6.69913 years. Parents of 76 patients reported withconsanguineous marriages while positive family history was reported in 31 patients. Eightytwo cases managed to ambulate with support of other persons while n= 14 patients weretotally non ambulatory. Symmetrical muscle involvement (lower limb followed by upper limb)was shown by 87 cases (including all males and n=22 females). Involvement of the lowerlimbs as a primary weakness followed by symmetrical and severe involvement of the upperlimb girdle and lower limb girdle suggested limb girdle muscle dystrophy (LGMD) in 9 (6F,3M)patients. Involvement of shoulders and upper limbs as a primary weakness followed by facialmuscles and infrequent involvement of lower limbs suggested fascioscapulohumeral muscledystrophy (FSHMD) in 2 cases. Involvement of upper limbs as a primary weakness followedby lower limbs with a moderate rise in CPK levels and mild phenotype was seen in two femalepatient which was quite suggestive of dysferlinopathy. Only two male patients reported onsetof weakness and ambulation loss before 1 year of age that may represent congenital muscledystrophy (CMD), dystroglycanopathy or congenital onset of LGMD (cLGMD). Mean serumCPK, LDH and Aldolase levels were 2376.6364 + 910.78963 U/L, 1030.7800 + 180.1620 U/Land 10.089 + 1.525 U/L respectively that demonstrated significant association (p&lt;0.05) withcharacteristic clinical features. Conclusion: Muscular dystrophies are not only restrictedto dystrophinopathies in our population. Apart from clinical and biochemical parameters,appropriate histological and/or gene mutation analysis is mandatory for precise classificationof these disorders.