OBJECTIVEThis study aims to explore the clinical applicability and relevance of glycosaminoglycan Chemical Exchange Saturation Transfer (gagCEST) for intervertebral disc.

METHODS25 subjects ranging in age from 24 yrs to 74 yrs were enrolled. gagCEST was acquired using a single-slice TSE sequence on a 3T. Saturation used a continuous rectangular RF pulse with B1=0.8 µT and a fixed duration time=1100 ms. Sagittal image was obtained firstly without saturation pulse, and then saturated images were acquired at 52 offsets ranging from ±0.125 to ±7 parts per million (ppm). MR T2 relaxivity map was acquired at the identical location. Six subjects were scanned twice to assess scan-rescan reproducibility.

RESULTSGagCEST intraclass correlation coefficient (ICC) of six subjects was 0.759 for nucleus pulposus (NP) and 0.508 for annulus fibrosus (AF). Bland-Altman plots showed NP had a mean difference of 0.10% (95% limits of agreement: -3.02% to 3.22%); while that of AF was 0.34% (95% limits of agreement: -2.28% to 2.95%). For the 25 subjects, gag CEST in NP decreased as disc degeneration increased, with a similar trend to T2 relaxivity. Gag CEST of AF showed a better correlation with disc degeneration than T2 relaxivity.

CONCLUSIONGagCEST in NP and AF decreased as disc degeneration increased, while gagCEST in AF showed a better correlation than T2 relaxivity.

Adult ; Aged ; Biomarkers ; analysis ; Case-Control Studies ; Female ; Glycosaminoglycans ; chemistry ; metabolism ; Humans ; Intervertebral Disc ; chemistry ; metabolism ; Intervertebral Disc Degeneration ; diagnosis ; metabolism ; Lumbar Vertebrae ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged

OBJECTIVEMucopolysaccharidosis type VI (MPS VI) or Maroteaux-Lamy syndrome is an autosomal recessive lysosomal storage disease caused by a deficiency of arylsulfatase B(ARSB), which is required in the degradation of dermatan sulfate and chondroitin sulfate. The deficiency of ARSB leads to an accumulation of dermatan sulfate and chondroitin sulfate in lysosomes and gross excretion in the urine.Few articles about clinical study and ARSB gene mutation analysis of Chinese MPS VI patients were published. This study aimed to explore the clinical features and characteristics of ARSB gene in Chinese children with MPS VI.

METHODThirteen children were diagnosed as MPS VI by ARSB enzyme activity determination during the period from 2009 to 2013. Their clinical features, radiological findings and urine glycosaminoglycan (GAG) levels were retrospectively reviewed. Direct sequencing was used to identify any mutation in the ARSB gene.

RESULTThirteen children were diagnosed at the average age of (3.9 ± 2.2) years with 6 male and 7 female. All of these children presented with severe form and onset at an early age of (1.5 ± 0.8) years.Other clinical features included coarse facies, short stature, skeleton deformity, corneal clouding, hepatosplenomegaly with normal intelligence. The radiological findings in all children were characteristic of dysostosis multiplex, like abnormal development of vertebral bodies of the spine, campylorrhachia and paddle-shaped widened ribs. The MRI in case 2 showed cervical cord compression and multiple cysts degeneration in the corona radiate, cella lateralis and callosum.High urine GAG levels were detected, (307.10 ± 112.14) mg/L (Normally below 70 mg/L) and (722.28 ± 245.68) µg/mg creatinine. The ARSB enzyme activity in leukocytes was low, (13.29 ± 6.22) nmol/(mg×h) [Normal range (47-169) nmol/(mg×h)] by fluorogenic assay and (0.24 ± 0.18) U/g [Normal range (1.01-11.47) U/g] by colorimetric assay. A total of 11 mutations were identified by molecular analysis, including seven previously reported mutations (p.L72R, p.G167R, p.G303E, p.F399L, p. T442M, p.Y255X and p.R327X) and four novel mutations (p.Y175D, p.S403X, p.S464X and large deletion including ex. 2, 3). The c.1197C>G (p.F399L) mutation was the most common mutation in this study (31%).

CONCLUSIONThe severe form of MPS VI is characterized by early onset and rapid illness progression. Both the radiological findings and increased urine GAG are important clues to diagnose MPS VI.Large decrease or absence of ARSB activity is diagnostic for MPS VI.Four novel mutations of ARSB gene were identified. The reported mutation c.1197C>G (p.F399L) was the hot-spot mutation in this study.

Bone and Bones ; diagnostic imaging ; pathology ; Brain ; pathology ; Child ; Child, Preschool ; Exons ; genetics ; Female ; Glycosaminoglycans ; urine ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Mucopolysaccharidosis VI ; diagnosis ; enzymology ; genetics ; Mutation ; N-Acetylgalactosamine-4-Sulfatase ; genetics ; metabolism ; Polymerase Chain Reaction ; Radiography ; Retrospective Studies ; Sequence Analysis, DNA

Mucopolysaccharidosis (MPS) III has 4 enzymatically distinct forms (A, B, C, and D), and MPS IIIC, also known as Sanfilippo C syndrome, is an autosomal recessive lysosomal storage disease caused by a deficiency of heparan acetyl-CoA:alpha-glucosaminide N-acetyltransferase (HGSNAT). Here, we report a case of MPS IIIC that was confirmed by molecular genetic analysis. The patient was a 2-yr-old girl presenting with skeletal deformity, hepatomegaly, and delayed motor development. Urinary excretion of glycosaminoglycan (GAG) was markedly elevated (984.4 mg GAG/g creatinine) compared with the age-specific reference range (<175 mg GAG/g creatinine), and a strong band of heparan sulfate was recognized on performing thin layer chromatography. HGSNAT enzyme activity in leukocytes was 0.7 nmol/17 hr/mg protein, which was significantly lower than the reference range (8.6-32 nmol/17 hr/mg protein). PCR and direct sequencing of the HGSNAT gene showed 2 mutations: c.234+1G>A (IVS2+1G>A) and c.1150C>T (p.Arg384*). To the best of our knowledge, this is the first case of MPS IIIC to be confirmed by clinical, biochemical, and molecular genetic findings in Korea.
Acetyltransferases/*genetics ; Asian Continental Ancestry Group/*genetics ; Base Sequence ; Child, Preschool ; Chromatography, Thin Layer ; Female ; Glycosaminoglycans/urine ; Heparitin Sulfate/chemistry/metabolism ; Humans ; Leukocytes/immunology/metabolism ; Mucopolysaccharidosis III/*diagnosis/genetics/radiography ; Mutation ; Republic of Korea ; Sequence Analysis, DNA

The periodontal ligament-derived mesenchymal stem cell is regarded as a source of adult stem cells due to its multipotency. However, the proof of chondrogenic potential of the cells is scarce. Therefore, we investigated the chondrogenic differentiation capacity of periodontal ligament derived mesenchymal stem cells induced by transforming growth factor (TGF)-β3 and bone morphogenetic protein (BMP)-6. After isolation of periodontal ligament stem cells (PDLSCs) from human periodontal ligament, the cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with 20% fetal bovine serum (FBS). A mechanical force initiated chondrogenic differentiation of the cells. For chondrogenic differentiation, 10 µg·L⁻¹ TGF-β3 or 100 µg∙L⁻¹ BMP-6 and the combination treating group for synergistic effect of the growth factors. We analyzed the PDLSCs by fluorescence-activated cell sorting and chondrogenesis were evaluated by glycosaminoglycans assay, histology, immunohistochemistry and genetic analysis. PDLSCs showed mesenchymal stem cell properties proved by FACS analysis. Glycosaminoglycans contents were increased 217% by TGF-β3 and 220% by BMP-6. The synergetic effect of TGF-β3 and BMP-6 were shown up to 281% compared to control. The combination treatment increased Sox9, aggrecan and collagen II expression compared with not only controls, but also TGF-β3 or BMP-6 single treatment dramatically. The histological analysis also indicated the chondrogenic differentiation of PDLSCs in our conditions. The results of the present study demonstrate the potential of the dental stem cell as a valuable cell source for chondrogenesis, which may be applicable for regeneration of cartilage and bone fracture in the field of cell therapy.
Adult Stem Cells ; physiology ; Aggrecans ; analysis ; Bone Morphogenetic Protein 6 ; pharmacology ; Cell Culture Techniques ; Cell Differentiation ; drug effects ; Cell Separation ; Chondrogenesis ; drug effects ; physiology ; Collagen Type II ; analysis ; Flow Cytometry ; Glycosaminoglycans ; analysis ; Humans ; Immunohistochemistry ; Mesenchymal Stromal Cells ; drug effects ; physiology ; Molar, Third ; cytology ; Periodontal Ligament ; cytology ; drug effects ; Reverse Transcriptase Polymerase Chain Reaction ; SOX9 Transcription Factor ; analysis ; Stress, Mechanical ; Tooth, Impacted ; pathology ; Transforming Growth Factor beta3 ; pharmacology

BACKGROUND: Impaired renal function may contribute to development of stroke and small vessel pathology in the brain. We investigated whether stroke subtype, initial stroke severity, early neurologic outcomes, time to cerebral infarction occurrence, and the presence of small vessel pathology in the brain are different between patients with end stage renal disease (ESRD) and those with renal transplantation (RT). METHODS: A total of 57 consecutive de novo RT patients (RT group) and 120 patients undergoing dialysis due to ESRD (ESRD group) who developed a first-ever acute cerebral infarction were enrolled. We compared stroke subtypes based on the Trial of Org 10172 in Acute Stroke Treatment classification, the presence of small vessel pathology (cerebral microbleed, leukoaraiosis and silent lacunar infarction) on MRI, stroke severity based on the National Institutes of Health Stroke Scale (NIHSS) and in-hospital mortality between the groups. RESULTS: The stroke subtypes, NIHSS scores at admission and in-hospital mortality were not different between the two groups. On multivariate analysis, the presence of high grade periventricular white matter changes tended to be more frequently detected in the ESRD group than the RT (P=0.078). The time from starting dialysis to stroke was longer in the RT group (129.9+/-60.9 months) than in the ESRD group (51.1+/-46.1 months). CONCLUSIONS: The stroke patterns, severity and short term outcomes were not different between RT and ESRD. The risk of cerebral infarction and high grade periventricular white matter changes may be reduced after RT in patients with ESRD.
Brain ; Cerebral Infarction ; Chondroitin Sulfates ; Dermatan Sulfate ; Dialysis ; Glycosaminoglycans ; Heparitin Sulfate ; Hospital Mortality ; Humans ; Kidney Failure, Chronic ; Kidney Transplantation ; Leukoaraiosis ; Multivariate Analysis ; National Institutes of Health (U.S.) ; Stroke

BACKGROUND AND OBJECTIVES: Advanced glycation end-products (AGEs) contribute to the development of atherosclerosis. We investigated whether serum AGEs are related to the presence or severity of coronary artery disease (CAD), and explored the association between serum AGEs and arterial stiffness according to diabetes status in patients suspected of having CAD. SUBJECTS AND METHODS: The measurement of serum AGEs and brachial-ankle pulse wave velocity (baPWV) were performed in 145 consecutive patients (63+/-9 years, 58% men) who received a coronary angiogram for evaluation of CAD. RESULTS: Forty-four diabetics and 101 non-diabetics were classified into three subgroups based on the number of diseased vessels with obstructive CAD: 0, 1, and 2 or more vessel diseases (VDs). Serum AGEs were significantly higher in diabetics with obstructive CAD than in those without obstructive CAD (2.16+/-0.29 vs. 1.85+/-0.29 mU/mL, p=0.010) and were significantly correlated with the number of VDs only in diabetics (r=0.504, p<0.001). Serum AGEs were not significantly correlated with baPWV in diabetics or non-diabetics. In receiver operating characteristics analysis, the cut-off value of serum AGEs as a predictor of obstructive CAD was 1.98 mU/mL, with 64% sensitivity and 63% specificity in diabetics. In multiple regression analysis, serum AGEs independently predicted obstructive CAD and were associated with the number of VDs in diabetics. CONCLUSION: Serum AGEs independently predict obstructive CAD and the severity of coronary atherosclerosis irrespective of arterial stiffness only in diabetics. Evaluation of PWV and serum AGEs together may be more effective to identify the risk of CAD in diabetic individuals.
Atherosclerosis ; Coronary Artery Disease ; Coronary Vessels ; Diabetes Mellitus ; Glycosaminoglycans ; Humans ; Pulse Wave Analysis ; ROC Curve ; Sensitivity and Specificity ; Vascular Stiffness

OBJECTIVE: Several studies showed that increased arterial stiffness is an independent risk factor for cardiovascular disease. Pulse wave velocity (PWV) is known as a marker for large vessel stiffness. Recent studies show that serum cystatin C is associated with PWV and may predict future cardiovascular events, even in subjects with normal renal function. However, there have been few studies for the relationship between cystatin C and arterial stiffness in patients with type 2 diabetes (T2DM). In this study, we investigated the relationship between serum cystatin C and branchial-ankle PWV in T2DM patients with normal renal function. METHODS: Patients with urinary albumin/creatinine ratio (ACR) higher than 300 microg albumin/mg creatinine, estimated glomerular filtration rate (eGFR) less than 60 mL/min were excluded. A total of 88 patients (47 male/41 female; age, 59+/-2 years; ACR, 33+/-5 microg/mg) were included. Doppler-derived aortic PWV and serum cystatin C were measured. RESULTS: Cystatin C is significantly related to age (r=0.51, P<0.001), hemoglobin A1c (r=-0.23, P<0.05), high density lipoprotein-cholesterol (r=-0.22, P<0.05), apoprotein A (r=-0.22, P<0.05), and eGFR (r=-0.56, P<0.001). Aortic PWV is significantly associated with age (r=0.29, P<0.01), cystatin C (r=0.33, P<0.005), and eGFR (r=-0.24, P<0.05). In multiple regression analysis, there is significant association between aortic PWV and serum cystatin C levels. CONCLUSION: Serum cystatin C is significantly associated with arterial stiffness in T2DM patients with normal renal function. Our results suggest that cystatin C could be a marker for early atherosclerosis in T2DM patients.
Apoproteins ; Atherosclerosis ; Cardiovascular Diseases ; Creatinine ; Cystatin C ; Diabetes Mellitus, Type 2 ; Glomerular Filtration Rate ; Glycosaminoglycans ; Hemoglobins ; Humans ; Pulse Wave Analysis ; Risk Factors ; Vascular Stiffness

BACKGROUND AND OBJECTIVES: The impact of multivessel coronary disease (MVD) with chronic total occlusion (CTO) on one-year mortality in patients with acute myocardial infarction (AMI) is not clearly known. We investigated the impact of MVD with concurrent CTO lesion on one-year mortality in patients with AMI. SUBJECTS AND METHODS: We studied 1008 consecutive patients who underwent coronary angiography between November 2005 and December 2008 with a diagnosis of AMI. RESULTS: Among 1008 patients, 432 patients (43%) had MVD, and 88 patients (8.7%) had CTO lesion. The one-year overall mortality was higher in patients with MVD than in patients with single vessel disease (SVD) (10.2% vs. 5.9%, p=0.012). However, the one-year overall mortality was not significantly higher in patients with CTO lesion than in patients without that lesion (12.5% vs. 7.3%, p=0.080). In multivariate analysis, independent predictors of one-year overall mortality were age older than 65 years {hazard ratio (HR) 2.41, 95% confidence interval (CI): 1.43 to 4.08}, Killip class > or =III (HR 3.59, 95% CI: 2.24 to 5.77), ST-elevation myocardial infarction (HR 2.45, 95% CI: 1.49 to 4.05) and MVD (HR 1.76, 95% CI: 1.07 to 2.89). CONCLUSION: Patients with MVD showed higher one-year mortality than patients with SVD. However, the presence of CTO was not an independent predictor of one-year mortality in this study that included patients with successfully revascularized CTO lesion.
Chronic Disease ; Coronary Angiography ; Coronary Disease ; Coronary Occlusion ; Glycosaminoglycans ; Humans ; Multivariate Analysis ; Myocardial Infarction ; Prognosis

Various kinds of glycosaminoglycans (GAGs) and proteoglycans (PGs) have been known to be involved in structural and space-filling functions, as well as many physiological regulations in skin. To investigate ultraviolet (UV) radiation-mediated regulation of GAGs and PGs in cultured human dermal fibroblasts, transcriptional changes of many types of PGs and GAG chain-synthesizing enzymes at 18 hr after 75 mJ/cm2 of UV irradiation were examined using quantitative real-time polymerase chain reaction methods. Hyaluronic acid synthase (HAS)-1, -2, and -3 and hyaluronidase-2 mRNA expressions were significantly increased by UV irradiation. Expressions of lumican, fibromodulin, osteoglycin, syndecan-2, perlecan, agrin, versican, decorin, and biglycan were significantly decreased by UV irradiation, while syndecan-1 was increased. Expressions of GAG chain-synthesizing glycosyltransferases, xylosyltransferase-1, beta1,3-glucuronyltransferase-1, beta1,4-galactosyltransferase-2, -4, exostosin-1, chondroitin polymerizing factor, and chondroitin sulfate synthase-3 were significantly reduced, whereas those of beta1,3-galactosyltransferase-6, beta1,4-galactosyltransferase-3, -7, beta-1,3-N-acetylglucosaminyltran sferase-2, and -7 were increased by UV irradiation. Heparanase-1 mRNA expression was increased, but that of heparanase-2 was reduced by UV irradiation. Time-course investigation of representative genes showed consistent results. In conclusion, UV irradiation may increase hyaluronic acid production through HAS induction, and decrease other GAG productions through downregulation of PG core proteins and GAG chain-synthesizing glycosyltransferases in cultured human dermal fibroblasts.
Cell Line ; Fibroblasts/metabolism/radiation effects ; Gene Expression Regulation/radiation effects ; Glucuronosyltransferase/genetics/radiation effects ; Glycosaminoglycans/*biosynthesis/chemistry ; Glycosyltransferases/genetics/*metabolism ; Humans ; Hyaluronic Acid/biosynthesis ; Hyaluronoglucosaminidase/genetics/radiation effects ; Polymerase Chain Reaction ; Proteoglycans/*biosynthesis/genetics/radiation effects ; RNA, Messenger/analysis/genetics ; Skin/*metabolism/radiation effects ; Transcription, Genetic/radiation effects ; *Ultraviolet Rays

PURPOSE: Mucopolysaccharidosis II (MPS II) is a lysosomal storage disorder caused by a deficiency of iduronate-2 sulfatase (IdS), which is involved in the degradation of glycosaminoglycan (GAG). In this study, the frequency of fasting hypoglycemia in patients with MPS II was investigated and changes in accumulation of glycogen and GAG in the hepatocytes of IdS-knockout (KO) mice were evaluated before and after recombinant IdS enzyme replacement therapy (ERT). MATERIALS AND METHODS: Plasma glucose levels were evaluated after an 8-hour fast in 50 patients with MPS II. The IdS-KO mice were divided into three groups (group 2; saline, group 3; 0.15 mg/kg of IdS, and group 4; 0.5 mg/kg of IdS); wild-type mice were included as controls (group 1). ERT was initiated intravenously at four weeks of age, and continued every week until 20 weeks of age. RESULTS: The mean glucose level after an 8-hour fast was 94.1 +/- 23.7 mg/dL in the patients with MPS II. Two (4%) out of 50 patients had fasting hypoglycemia. For the mice, GAG in the lysosomes nearly disappeared and glycogen particles in the cytoplasm were restored to the normal range in group 4. CONCLUSION: Glucose metabolism in patients with MPS II appeared to function well despite hepatocytic GAG accumulation and hypothetical glycogen depletion. A higher dose of IdS infusion in MPS II mice led to disappearance of lysosomal GAG and restoration of glycogen to the cytoplasm of hepatocytes.
Animals ; Blood Glucose/analysis ; Enzyme Replacement Therapy/methods ; Glycogen/*analysis ; Glycosaminoglycans/*analysis ; Hepatocytes/chemistry/*enzymology ; Humans ; Hypoglycemia/enzymology/physiopathology ; Iduronate Sulfatase/genetics/*physiology ; Liver/ultrastructure ; Mice ; Mice, Knockout ; Microscopy, Electron ; Mucopolysaccharidosis II/blood/enzymology/physiopathology