2.Dual Roles of Ligamentum Flavum for Spinal Fusion: As an Osteoinductive Agent and Carrier for Ex-vivo Gene Transfer.
Seong Hwan MOON ; Hyang KIM ; Un Hye KWON ; Keong Hee KIM ; Hong Ki YOUN ; Hak Sun KIM ; Soo Bong HAHN ; Hwan Mo LEE
Journal of Korean Society of Spine Surgery 2003;10(1):1-7
STUDY DESIGN: An in-vitro experiment using human ligamentum flavum (LF) and the adnovirus-BMP-2 construct, Ad/BMP-2. OBJECTIVES: To determine the dual roles of LF as an osteoinductive agent and carrier for ex-vivo gene transfer. SUMMARY OF LITERATURE REVIEW: LF is known to have osteogenic potential. Pathologically, ossified LF may cause myelopathy and radiculopathy. BMP-2 is known as an important factor in the differentiation, and maintenance, of osteoblast phenotypes. Ex-vivo gene transfer, using human LF for spinal fusion, has never been attempted before. MATERIALS AND METHODS: The LF cells were cultured from the degenerated LF of spinal stenosis patients. An adenovirus construct, containing BMP-2 cDNA (Ad/BMP-2), was also produced. The LF cell cultures were exposed to the adenoviral construct. The Osteocalcin expression was analysed by Western blot analysis. The osteocalcin and BMP-2 mRNA expressions were analysed by RT-PCR. Bone formation was assessed by alkaline phosphatase and Von Kossa stains. RESULTS: The LF cell cultures, with Ad/BMP-2, showed transgene expression in the Western blot analysis. Also, the cultures exhibited the mRNA expressions of both osteocalcin and BMP-2, in a dose-dependent manner. The LF cultures, with Ad/BMP-2, demonstrated alkaline phosphatase expression and bone nodule formations from the Von Kossa staining. CONCLUSION: The genetically modified LF strongly induced osteogenesis, which can be used during a spinal fusion, as an osteoinductive agent and carrier, for ex-vivo gene transfer.
Adenoviridae
;
Alkaline Phosphatase
;
Blotting, Western
;
Cell Culture Techniques
;
Coloring Agents
;
DNA, Complementary
;
Humans
;
Ligamentum Flavum*
;
Osteoblasts
;
Osteocalcin
;
Osteogenesis
;
Phenotype
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Radiculopathy
;
RNA, Messenger
;
Spinal Cord Diseases
;
Spinal Fusion*
;
Spinal Stenosis
;
Transgenes
3.Serum Globotriaosylceramide Assay as a Screening Test for Fabry Disease in Patients with ESRD on Maintenance Dialysis in Korea.
Jeong Yup KIM ; Young Youl HYUN ; Ji Eun LEE ; Hye Ran YOON ; Gu Hwan KIM ; Han Wook YOO ; Seong Tae CHO ; No Won CHUN ; Byoung Chunn JEOUNG ; Hwa Jung KIM ; Keong Wook KIM ; Seong Nam KIM ; Yung A KIM ; Hyun Ah LEE ; Jong Young LEE ; Yung Chun LEE ; Hun Kwan LIM ; Keong Sik OH ; Seong Hwan SON ; Beong Hee YU ; Kyeong So WEE ; Eun Jong LEE ; Young Ki LEE ; Jung Woo NOH ; Seung Jung KIM ; Kyu Bok CHOI ; Suk Hee YU ; Heui Jung PYO ; Young Joo KWON
The Korean Journal of Internal Medicine 2010;25(4):415-421
BACKGROUND/AIMS: Fabry disease is an X-linked recessive and progressive disease caused by alpha-galactosidase A (alpha-GaL A) deficiency. We sought to assess the prevalence of unrecognized Fabry disease in dialysis-dependent patients and the efficacy of serum globotriaosylceramide (GL3) screening. METHODS: A total of 480 patients of 1,230 patients among 17 clinics were enrolled. Serum GL3 levels were measured by tandem mass spectrometry. Additionally, we studied the association between increased GL3 levels and cardiovascular disease, cerebrovascular disease, or left ventricular hypertrophy. RESULTS: Twenty-nine patients had elevated serum GL3 levels. The alpha-GaL A activity was determined for the 26 patients with high GL3 levels. The mean alpha-GaL A activity was 64.6 nmol/hr/mg (reference range, 45 to 85), and no patient was identified with decreased alpha-GaL A activity. Among the group with high GL3 levels, 15 women had a alpha-GaL A genetics analysis. No point mutations were discovered among the women with high GL3 levels. No correlation was observed between serum GL3 levels and alpha-GaL A activity; the Pearson correlation coefficient was 0.01352 (p = 0.9478). No significant correlation was observed between increased GL3 levels and the frequency of cardiovascular disease or cerebrovascular disease. CONCLUSIONS: Fabry disease is very rare disease in patients with end-stage renal disease. Serum GL3 measurements as a screening method for Fabry disease showed a high false-positive rate. Thus, serum GL3 levels determined by tandem mass spectrometry may not be useful as a screening method for Fabry disease in patients with end stage renal disease.
Adult
;
Aged
;
Fabry Disease/blood/*diagnosis
;
Female
;
Humans
;
Kidney Failure, Chronic/blood/*therapy
;
Male
;
Middle Aged
;
*Renal Dialysis
;
Trihexosylceramides/*blood
;
alpha-Galactosidase/genetics/metabolism