Pseudohypoparathyroidism is a medical disorder characterized by a complex disorder of renal resistance to parathyroid hormone and the mechanism underlying the disease is still unclear. The authors described two cases of pseudohypoparathyroidism in sibling,who had metabolic anomalies(hypocalcemia and hyperphosphatemia, high circulatin immunoreactive PTH)and basal ganglia calcification. Bilateral basal ganglia calcifications, which was not visible on plain skull film, was detected by CT scan of brain MRI. We report these cases with a review of related literatures.
Basal Ganglia ; Brain ; Humans ; Hyperphosphatemia ; Magnetic Resonance Imaging ; Parathyroid Hormone ; Pseudohypoparathyroidism* ; Siblings* ; Skull ; Tomography, X-Ray Computed

STUDY DESIGN: This is an in-vitro experiment using rabbit intervertebral disc (IVD) cells and growth factors. OBJECTIVES: We wanted to determine the effect of types I, and II atelocollagen and growth factor gene therapy for matrix regeneration of rabbit IVD cells. SUMMARY OF THE LITERATURE REVIEW: Adenovirus-medicated growth factor gene therapy is efficient for matrix regeneration of the IVD. Atellocollagen has provided a favorable environment for matrix synthesis. However, a combined approach using gene and cell therapy in an atelocollagen scaffold has not yet been attempted. MATERIALS AND METHODS: Rabbit IVD cells were transduced with Ad/TGF-beta1 and Ad/BMP-2. The cells were then implanted to the atelocollagen scaffold. The [methyl-3H]thymidine incorporation for DNA synthesis and the [35S]sulfur incorporation for proteoglycan synthesis were measured. RT-PCR was performed for assessing the aggrecan, collagen type I, collagen type II and osteocalcin mRNA expressions. RESULTS: The rabbit IVD cells with Ad/TGF-beta1 and that were cultured in type I atelocollagen showed a 130% increase in new proteoglycan synthesis, while the rabbit IVD cells with Ad/TGF-beta1 and that were cultured in type II atelocollagen showed a 180% increase in new proteoglycan synthesis (p<0.05). The rabbit IVD cells with Ad/BMP-2 and that were cultured in type I atelocollagen showed a 70% increase in new proteoglycan synthesis, while the rabbit IVD cells with Ad/BMP-2 and that were cultured in type II atelocollagen showed a 95% increase (p<0.05). Rabbit IVD cells with Ad/TGF-beta1 and Ad/BMP-2 and that were cultured in type I and II atelocollagen demonstrated increased collagen type I and II mRNA expressions without an osteocalcin mRNA expression (p<0.05). CONCLUSION: Cell and gene therapy in an atelocollagen scaffold provided a efficient mechanism for chondrogenic matrix regeneration of rabbit IVD cells.
Aggrecans ; Collagen ; Collagen Type I ; Collagen Type II ; DNA ; Genetic Therapy ; Intercellular Signaling Peptides and Proteins ; Intervertebral Disc ; Osteocalcin ; Proteoglycans ; Regeneration ; RNA, Messenger ; Tissue Engineering ; Tissue Therapy ; Transforming Growth Factor beta1

Our recent study has provided that the in vitro SEC-induced proliferation of bovine T cells is preceded by a period of a non-proliferative immunoregulation of T cells that may be associated with cytokine production regulated by type 1 or type 2 T cells. Inversion of CD4+:CD8+ T cell ratio and induction of CD8+T cells with immunoregulatory activity could increase the probability of intracellular survival of Staphylococcus aureus (S. aureus). The increase of activated CD8+(ACT2+ BoCD8+) T cells in cows with mastitis caused by S. aureus may be associated with immune-regulatory function in the bovine mammary gland. The difference and similarity between bovine activated CD8+ T cells (CD8+ CD26+)and well-established human CD4+ CD25+ T regulatory (Tr)cells may help to reveal their unique immune regulatory system in the host infected with S. aureus.
Animals ; Cattle ; Cell Proliferation ; Female ; Lymphocyte Activation/immunology ; Mastitis, Bovine/*immunology/microbiology ; Staphylococcal Infections/immunology/*veterinary ; Staphylococcus/*immunology ; *Superantigens ; T-Lymphocytes/*immunology

STUDY DESIGN: An in vitro experiment. OBJECTIVES: To assess the effect of pulsed sinusoidal EMF on human intervertebral disc (IVD) cells. LITERATURE REVIEW SUMMARY: Electromagnetic field (EMF) is known to modify some relevant physiological parameters of cells cultured in vitro, such as proliferation, synthesis, secretion of growth factors and transcription. EMF induces bone formation in delayed, non union and spinal fusion models. Also, the exposure of EMF has been shown to protect against the hazardous effect of smoking in the rabbit IVD. MATERIALS AND METHODS: Human IVD cells were three-dimensionally cultured in alginate beads and exposed to a 650 omega, 1.8millitesla magnetic flux density, 60Hz sinusoidal wave of EMF. The cultures were divided into the control and EMF groups, with various exposure times. The cytotoxicity, and DNA and proteoglycan syntheses were measured by the MTT assay, and [3H]-thymidine and [35S]-sulfate incorporation, respectively. RT-PCRs were performed for aggrecan, and collagen types I and II mRNA expressions. RESULTS: There was no recognizable cytotoxicity in the EMF group, but cellular proliferation was stimulated (p<0.05). Newly synthesized proteoglycan, normalized by DNA synthesis, was decreased in the EMF group (p<0.05) as were the expressions of aggrecan (48hour exposure) and type II collagen (72 hours exposure) mRNA compared to the control group. CONCLUSIONS: EMF seems to be hazardous in the synthesis of the chondrogenic matrix, while marginally beneficial in the cellular proliferation of human IVD cells.
Aggrecans ; Cell Proliferation ; Collagen ; Collagen Type II ; DNA ; Electromagnetic Fields* ; Humans* ; Intercellular Signaling Peptides and Proteins ; Intervertebral Disc* ; Magnets* ; Osteogenesis ; Proteoglycans ; RNA, Messenger ; Smoke ; Smoking ; Spinal Fusion

STUDY DESIGN: An in vitro experiment. OBJECTIVES: To assess the effect of pulsed sinusoidal EMF on human intervertebral disc (IVD) cells. LITERATURE REVIEW SUMMARY: Electromagnetic field (EMF) is known to modify some relevant physiological parameters of cells cultured in vitro, such as proliferation, synthesis, secretion of growth factors and transcription. EMF induces bone formation in delayed, non union and spinal fusion models. Also, the exposure of EMF has been shown to protect against the hazardous effect of smoking in the rabbit IVD. MATERIALS AND METHODS: Human IVD cells were three-dimensionally cultured in alginate beads and exposed to a 650 omega, 1.8millitesla magnetic flux density, 60Hz sinusoidal wave of EMF. The cultures were divided into the control and EMF groups, with various exposure times. The cytotoxicity, and DNA and proteoglycan syntheses were measured by the MTT assay, and [3H]-thymidine and [35S]-sulfate incorporation, respectively. RT-PCRs were performed for aggrecan, and collagen types I and II mRNA expressions. RESULTS: There was no recognizable cytotoxicity in the EMF group, but cellular proliferation was stimulated (p<0.05). Newly synthesized proteoglycan, normalized by DNA synthesis, was decreased in the EMF group (p<0.05) as were the expressions of aggrecan (48hour exposure) and type II collagen (72 hours exposure) mRNA compared to the control group. CONCLUSIONS: EMF seems to be hazardous in the synthesis of the chondrogenic matrix, while marginally beneficial in the cellular proliferation of human IVD cells.
Aggrecans ; Cell Proliferation ; Collagen ; Collagen Type II ; DNA ; Electromagnetic Fields* ; Humans* ; Intercellular Signaling Peptides and Proteins ; Intervertebral Disc* ; Magnets* ; Osteogenesis ; Proteoglycans ; RNA, Messenger ; Smoke ; Smoking ; Spinal Fusion

STUDY DESIGN: In-vitro experiment. OBJECTIVES: To determine the effect of bone morphogenetic protein-2 in the osteogenesis of human ligamentum flavum cells and test the feasibility of gene transfer to these cells. SUMMARY OF LITERATURE REVIEW: Bone morphogenetic protein-2 (BMP-2) is known to be an important factor in the differentiation and maintenance of the osteoblastic phenotype. Tissue engineering for osteogenesis in ligamentum flavum by BMP-2 and gene transfer has not been previously studied. MATERIALS AND METHODS: Ligmentum flavum cells were harvested and cultured from surgical patients with spinal stenosis. BMP-2 was produced by transfecting pcDNA3.1/Hygro/BMP-2 into CHO cells using Lipofectamine 2000. Adenovirus-lacZ (Ad/lacZ) was also produced, and administered with BMP-2 to cell culture. The expression of lacZ was analyzed by X-gal staining. Bone formation was assessed by alkaline phosphatase, von Kossa, and alizarin Red-S staining, and the expression of osteocalcin was determined immunocytochemically. RESULTS: Ligamentum flavum cell cultures with Ad/lacZ showed marker gene expression. BMP-2 induced osteogenesis in ligamentum flavum cells as evidenced by alkaline phosphatase, von Kosa, and alizarin Red-S staining. Also, cell culture with BMP-2 showed strong positivity with osteocalcin by immunocytochemistry. CONCLUSION: BMP-2 more strongly induced the osteogenesis of ligamentum flavum, and also its gene transfer to ligamentum flavum was found to be feasible. These results may open a new era of ligamentum flavum tissue engineering.
Adenoviridae ; Alkaline Phosphatase ; Animals ; Cell Culture Techniques ; CHO Cells ; Cricetinae ; DNA, Complementary* ; Gene Expression ; Humans ; Immunohistochemistry ; Ligamentum Flavum* ; Osteoblasts ; Osteocalcin ; Osteogenesis* ; Phenotype ; Spinal Stenosis ; Tissue Engineering*

Antidiuretic action of oxytocin is confirmed by in vitro study using with rat IMCD. Vasopressin is elevated in edematous disorders and may play a pathogenetic role in the formation of edema. If oxytocin plays a sirnilar role to vasopressin in water disturbances in human, oxytocin may change as the same way as vasopressin. To verify a role of oxytocin in the regulation of water balance in human, we measured plasma and urine oxytocin with vasopressin by radioimmunoassay in thirteen patients with generalized edema (8 nephrotic syndrome, 3 liver cirrhosis, 2 acute renal failure) before and after control of edema. And they were compared them with those of seven normal controls. Plasma oxytocin level correlated with plasma vasopressin level (r=0.543: P<0.05) and urinary oxytocin level correlated linearly with urinary vaso-pressin (r=0.983, P<0.01). After control of edema, body weight of patients decreased from 65+/- 2 to 58+/-2kg and fractional excretion of sodium decreased from 3.3+/-1.1 to 1.2+/-0.696 (P<0.05). There were no significant changes in serum and urine Na, osmolality, free water clearance, plasma renin activity, aldosterone and norepinephrine. In conclusion, oxytocin was elevated in edematous disorders, and may participate in formation of edema similar to vasopressin.
Aldosterone ; Animals ; Body Weight ; Edema ; Humans ; Liver Cirrhosis ; Nephrotic Syndrome ; Norepinephrine ; Osmolar Concentration ; Oxytocin* ; Plasma ; Radioimmunoassay ; Rats ; Renin ; Sodium ; Vasopressins* ; Water*

STUDY DESIGN: In-vitro experiments using human mesenchymal stem cells (MSCs), intervertebral disc (IVD) cells and type 5 adenovirus/transforming growth factor-beta1 construct (Ad/TGF-beta1). OBJECTIVES: To determine the effect of MSC-based gene therapy for matrix regeneration of IVD cells. SUMMARY OF LITERATURE REVIEW: MSCs are known to be multipotent in tissue regeneration. In degeneration of IVD, cellular replacement with genetic modification other than that of IVD cells may prove an enhanced mechanism for the regeneration of MATERIALS AND METHODS: MSCs and IVD cells were cultured and an adenovirus construct containing TGF-beta1 cDNA (Ad/TGF-beta1) was also produced. In the first step, the MSCs were transduced with Ad/TGF-beta1, then mixed with IVD cells in various proportions and three dimensionally cultured. [methyl-(3)H]Thymidine and [(35)S]Sulfur incorporation for DNA and proteoglycan synthesis, respectively, were measured. RT-PCR was performed to assess the aggrecan and collagen types I and II mRNA RESULTS: Mixed cultures of MSC and IVD cells showed relatively similar amounts of newly synthesized proteoglycan compared with cultures of IVD cells only. In mixed cultures transduced with Ad/TGF-beta1, there were significant decreases in newly synthesized proteoglycan with increasing the proportions of MSCs, which was also found with the aggrecan and collagen type II mRNA expressions. However, the collagen type I mRNA expression increased with increased proportions of MSCs transduced with Ad/TGF-beta1. CONCLUSION: Cell therapy with MSCs and IVD cells provided a mechanism for cellular augmentation. However, MSC-based gene therapy coupled with IVD cells did not maintain a chondrogenic phenotype.
Adenoviridae ; Aggrecans ; Cell- and Tissue-Based Therapy ; Collagen ; Collagen Type I ; Collagen Type II ; DNA ; DNA, Complementary ; Genetic Therapy* ; Humans ; Intervertebral Disc ; Mesenchymal Stromal Cells ; Phenotype ; Proteoglycans ; Regeneration ; RNA, Messenger ; Stem Cells* ; Transforming Growth Factor beta1

PURPOSE: The purpose of this study is to investigate the mechanism of cellular proliferation of electromagnetic field (EMF) on human intervertebral disc (IVD) cells. MATERIALS AND METHODS: Human IVD cells were cultured three-dimensionally in alginate beads. EMF was exposed to IVD cells with 650Omega, 1.8 millitesla magnetic flux density, 60 Hz sinusoidal wave. Cultures were divided into a control and EMF group. Cytotoxicity, DNA synthesis and proteoglycan synthesis were measured by MTT assay, [3H]-thymidine, and [35S]-sulfate incorporation. To detect phenotypical expression, reverse transcription-polymerase chain reactions (RT-PCR) were performed for aggrecan, collagen type I, and type II mRNA expression. To assess action mechanism of EMF, IVD cells were exposed to EMF with NG-Monomethyl-L-arginine (NMMA) and acetylsalicylic acid (ASA). RESULTS: There was no cytotoxicity in IVD cells with the EMF group in MTT assay. Cellular proliferation was observed in the EMF group (p < 0.05). There was no difference in newly synthesized proteoglycan normalized by DNA synthesis between the EMF group and the control. Cultures with EMF showed no significant change in the expression of aggrecan, type I, and type II collagen mRNA compared to the control group. Cultures with NMMA (blocker of nitric oxide) or ASA (blocker of prostaglandin E2) exposed to EMF demonstrated decreased DNA synthesis compared to control cultures without NMMA or ASA (p < 0.05). CONCLUSION: EMF stimulated DNA synthesis in human IVD cells while no significant effect on proteoglycan synthesis and chondrogenic phenotype expressions. DNA synthesis was partially mediated by nitric oxide and prostaglandin E2. EMF can be utilized to stimulate proliferation of IVD cells, which may provide efficient cell amplification in cell therapy to degenerative disc disease.
Adult ; Aspirin/pharmacology ; Cell Proliferation/*radiation effects ; Collagen/metabolism ; Dinoprostone/metabolism ; *Electromagnetic Fields ; Enzyme Inhibitors/pharmacology ; Female ; Humans ; Intervertebral Disk/*pathology/radiation effects ; Male ; Middle Aged ; Nitric Oxide/metabolism ; Tetrazolium Salts/pharmacology ; Thiazoles/pharmacology ; omega-N-Methylarginine/pharmacology

Ael is a rare blood type which has the least amount of A antigen among A subgroups. It can be detected by special tests performed to resolve the discrepancy between red cell and serum typing in routine serological typing. The presence of A antigen on Ael red cell is demonstrable only by adsorption and elution tests. An Ael individual does not secret A substance in the saliva and may have anti-A antibody in the serum which is usually less reactive with the reagent red cells than anti-B antibody. In Korea, Ael02 has been reported more frequently than other Ael alleles. We report a case of Ael02/O04 who presented as typical phenotype O with strong anti-A and anti-B antibodies and no A antigen detected even by adsorption and elution tests. The case has been proved to be Ael02/O04 by direct sequencing analysis. In individuals with history of discrepancies in the results of ABO phenotyping, ABO genotyping is needed for an accurate evaluation of their blood type.
ABO Blood-Group System/classification/*genetics ; Alleles ; Child ; Genotype ; Heterozygote ; Humans ; Male ; Pedigree ; Phenotype ; Sequence Analysis, DNA