Xenoantigen, an αGal epitope-expression construct driven by the hTERT-promoter, specifically kills human pancreatic cancer cell line

BACKGROUND: We previously reported the usefulness of the αGal epitope as a target molecule for gene therapy against cancer. To induce cancer cell specific transcription of the αGal epitope, an expression vector which synthesizes the αGal epitope under the control of a promoter region of the human telomerase reverse transcriptase (hTERT), NK7, was constructed. METHODS: NK7 was transfected into a human pancreatic carcinoma cell line, MIA cells, and telomerase-negative SUSM-1 cells served controls. Expression of the αGal epitope was confirmed by flow cytometry using IB4 lectin. The susceptibility of transfected MIA cells to human natural antibodies, was examined using a complement-dependent cytotoxic cross-match test (CDC) and a flow cytometry using annexin V. RESULTS: The αGal epitope expression was detected only on the cell surfaces of NK7-transfected MIA cells, i.e., not on naive MIA cells or telomerase negative SUSM-1 cells. The CDC results indicated that MIA cells transfected with NK7 are susceptible to human natural antibody-mediated cell killing, and the differences, as compared to NK-7 transfected telomerase negative SUSM-1 cells or telomerase positive naïve MIA cells, were statistically significant. The flow cytometry using annexin V showed a higher number of the apoptotic cells in NK-7 transfected MIA cells than in naïve MIA cells. CONCLUSIONS: The results suggest that αGal epitope-expression, under the control of the hTERT-promoter, may be useful in cancer specific gene therapy

Glycated albumin suppresses glucose-induced insulin secretion by impairing glucose metabolism in rat pancreatic β-cells

<p>Abstract</p> <p>Background</p> <p>Glycated albumin (GA) is an Amadori product used as a marker of hyperglycemia. In this study, we investigated the effect of GA on insulin secretion from pancreatic β cells.</p> <p>Methods</p> <p>Islets were collected from male Wistar rats by collagenase digestion. Insulin secretion in the presence of non-glycated human albumin (HA) and GA was measured under three different glucose concentrations, 3 mM (G3), 7 mM (G7), and 15 mM (G15), with various stimulators. Insulin secretion was measured with antagonists of inducible nitric oxide synthetase (iNOS), and the expression of iNOS-mRNA was investigated by real-time PCR.</p> <p>Results</p> <p>Insulin secretion in the presence of HA and GA was 20.9 ± 3.9 and 21.6 ± 5.5 μU/3 islets/h for G3 (<it>P </it>= 0.920), and 154 ± 9.3 and 126.1 ± 7.3 μU/3 islets/h (<it>P </it>= 0.046), for G15, respectively. High extracellular potassium and 10 mM tolbutamide abrogated the inhibition of insulin secretion by GA. Glyceraldehyde, dihydroxyacetone, methylpyruvate, GLP-1, and forskolin, an activator of adenylate cyclase, did not abrogate the inhibition. Real-time PCR showed that GA did not induce iNOS-mRNA expression. Furthermore, an inhibitor of nitric oxide synthetase, aminoguanidine, and NG-nitro-L-arginine methyl ester did not abrogate the inhibition of insulin secretion.</p> <p>Conclusion</p> <p>GA suppresses glucose-induced insulin secretion from rat pancreatic β-cells through impairment of intracellular glucose metabolism.</p

Expression of hepcidin mRNA is uniformly suppressed in hepatocellular carcinoma

<p>Abstract</p> <p>Background</p> <p>The present study evaluated the expression of hepcidin mRNA in hepatocellular carcinoma (HCC).</p> <p>Methods</p> <p>Samples of cancerous and non-cancerous liver tissue were taken from 40 patients with HCC who underwent hepatectomy. Expression of hepcidin mRNA was evaluated by real-time PCR, and compared in tumors differing in their degree of differentiation, number of tumors, and vessel invasion. Correlations between hepcidin expression and the interval until HCC recurrence, and the serum concentration of hepcidin were evaluated, together with the expression of mRNAs for other iron metabolism molecules, ferroportin and transferrin receptor 2 (Trf2).</p> <p>Results</p> <p>Hepcidin mRNA expression in non-cancerous and cancerous tissues was 1891.8 (32.3–23187.4) and 53.4 (1.9–3185.8), respectively (<it>P </it>< 0.0001). There were no significant differences in hepcidin expression among tumors differing in their degree of differentiation, number of tumors, or vessel invasion. There was no significant correlation between hepcidin expression and the interval until HCC recurrence. The serum concentration of hepcidin-25 was not correlated with hepcidin-mRNA expression. Finally, there were no significant differences in the expression of mRNA for ferroportin and Trf2 between cancerous and non-cancerous tissues.</p> <p>Conclusion</p> <p>Expression of hepcidin mRNA is strikingly suppressed in cancerous, but not in non-cancerous tissues, in patients with HCC, irrespective of ferroportin or Trf2 expression. Uniform suppression of hepcidin may be linked to the development of HCC.</p

Strategy for reduction of medical costs and growth of the healthcare industry after establishment of the national health insurance system in Japan: lessons for the Indonesian health care reform system -II-

Japan has long operated a government-based health insurance system covering the entire population, and has been focusing on strategies for balancing viable national health care services with the need to minimize national health care expenditure. Hemodialysis (HD) is a representative form of medical treatment that is expected to expand in Indonesia in the near future, and which will require sustained financial support from the national health insurance system. In this report, consecutive to our previous one, we describe how the Japanese government has attempted to minimize medical expenditure in the past, focusing especially on HD.</p

Strategy for reduction of medical costs and growth of the healthcare industry after establishment of the national health insurance system in Japan: lessons for the Indonesian health care reform system

AbstractJapan has long operated a government-based health insurance system covering the entire population, and has been focusing on strategies for balancing viable national health care services with the need to minimize national health care expenditure. Hemodialysis (HD) is a representative form of medical treatment that is expected to expand in Indonesia in the near future, and which will require sustained financial support from the national health insurance system. In this report, consecutive to our previous one, we describe how the Japanese government has attempted to minimize medical expenditure in the past, focusing especially on HD.Keywords: health care reform, medical cost, national health insurance3 hlm

慢性期移植腎機能低下例におけるIV型コラーゲンの分布の変化に関する検討

IV型コラーゲンは基底膜の主要構成成分であり,生体内に広く分布している.移植後急性期腎機能低下例においては,腎臓のIV型コラーゲン分布に変化が認められる.今回我々は移植腎機能廃絶の主原因である慢性期移植腎機能低下症(chronic allograft nephropathy: CAN)におけるIV型コラーゲンの変化について検討した.当院において施行された腎移植患者を,移植腎機能に基づいて以下の群に分類し,組織所見について検討した.なおCAN症例は,光学顕微鏡所見で診断された症例を用いた.Group A(n=5):腎機能正常群,Group B(n=10):急性期移植腎機能低下群,Group C(n=35): CAN症例で血清クレアチニン値2～4mg/dl,Group D(n=15): CANで血清クレアチニン値>4mg/dl.各症例におけるIV型コラーゲン分布について,IV型コラーゲンのα1鎖に対するモノクローナル抗体であるJK199,JK132を用いて,蛍光顕微鏡により検討した.Group AではJK199,JK132ともにメサンギウム基質(MM),ボーマン嚢基底膜(BBM),尿細管基底膜(TBM)に対する反応のみを認めた.Group Bでは,JK199はMM,BBM,TBMに加え,糸球体基底膜(GBM)および腎間質(INS)に対する反応を認めた.JK132の反応性はGroup Aと同様であった.一方,Group CおよびGroup Dでは,JK199においてはMM,GBM,BBM,TBM,INSに対する反応性が増強していた.JK132についてはMM,BBM,TBMに加え,GBMおよびINSにおいても反応が認められた.また,JK199およびJK132の反応性はGroup CよりもGroup Dにおいて強い傾向が認められた.GBMおよびINSにおけるJK132の反応陽性化はCANに特異的な所見であることが示唆された.Previously, we demonstrated the altered formation of collagen IV, which is the main constituent of the basement membrane, in renal allografts by staining with two monoclonal antibodies against the α1 chain of collagen IV. In the present study, we investigated the alteration of collagen IV in chronic allograft nephropathy (CAN), which is an irreversible change that can occur in renal allografts. Biopsy specimens of normal kidneys (Group A: n=5) and acute rejection (Group B: n=10) were studied as controls. Fifty biopsy specimens from 41 patients who had been diagnosed as having CAN were divided into two groups, according to renal function: Group C (n=35), sCr 2～4 mg/dl, and Group D (n=15), sCr>4 mg/dl. Two monoclonal antibodies, JK199 and JK132, those recognize the α1 chain of collagen IV were used. In Group A, JK199 reacted with the glomerular basement membrane (GBM), the mesangial matrix (MM), the basement membrane of Bowman's capsule (BBM) and the tubular basement membrane (TBM). JK132 only reacted with the MM, BBM and TBM. In Group B, JK199 reacted with GBM, MM, BBM, TBM and the interstitium (INS). JK132 only reacted with MM, BBM and TBM. In Group C and Group D, JK199 and JK132 reacted universally with GBM and INS in addition to MM, BBM and TBM. The intensity of the reaction was higher in Group D than in Group C. Thus, the reactivity of JK132 with GBM and INS was a unique finding for the CAN specimens. These results suggest that collagen IV is upregulated in CAN and the reactivity of JK132 in GBM and INS may represent the point of irreversible dysfunction of renal allografts

慢性期移植腎機能低下例におけるIV型コラーゲンの分布の変化に関する検討

IV型コラーゲンは基底膜の主要構成成分であり,生体内に広く分布している.移植後急性期腎機能低下例においては,腎臓のIV型コラーゲン分布に変化が認められる.今回我々は移植腎機能廃絶の主原因である慢性期移植腎機能低下症(chronic allograft nephropathy: CAN)におけるIV型コラーゲンの変化について検討した.当院において施行された腎移植患者を,移植腎機能に基づいて以下の群に分類し,組織所見について検討した.なおCAN症例は,光学顕微鏡所見で診断された症例を用いた.Group A(n=5):腎機能正常群,Group B(n=10):急性期移植腎機能低下群,Group C(n=35): CAN症例で血清クレアチニン値2～4mg/dl,Group D(n=15): CANで血清クレアチニン値>4mg/dl.各症例におけるIV型コラーゲン分布について,IV型コラーゲンのα1鎖に対するモノクローナル抗体であるJK199,JK132を用いて,蛍光顕微鏡により検討した.Group AではJK199,JK132ともにメサンギウム基質(MM),ボーマン嚢基底膜(BBM),尿細管基底膜(TBM)に対する反応のみを認めた.Group Bでは,JK199はMM,BBM,TBMに加え,糸球体基底膜(GBM)および腎間質(INS)に対する反応を認めた.JK132の反応性はGroup Aと同様であった.一方,Group CおよびGroup Dでは,JK199においてはMM,GBM,BBM,TBM,INSに対する反応性が増強していた.JK132についてはMM,BBM,TBMに加え,GBMおよびINSにおいても反応が認められた.また,JK199およびJK132の反応性はGroup CよりもGroup Dにおいて強い傾向が認められた.GBMおよびINSにおけるJK132の反応陽性化はCANに特異的な所見であることが示唆された.Previously, we demonstrated the altered formation of collagen IV, which is the main constituent of the basement membrane, in renal allografts by staining with two monoclonal antibodies against the α1 chain of collagen IV. In the present study, we investigated the alteration of collagen IV in chronic allograft nephropathy (CAN), which is an irreversible change that can occur in renal allografts. Biopsy specimens of normal kidneys (Group A: n=5) and acute rejection (Group B: n=10) were studied as controls. Fifty biopsy specimens from 41 patients who had been diagnosed as having CAN were divided into two groups, according to renal function: Group C (n=35), sCr 2～4 mg/dl, and Group D (n=15), sCr>4 mg/dl. Two monoclonal antibodies, JK199 and JK132, those recognize the α1 chain of collagen IV were used. In Group A, JK199 reacted with the glomerular basement membrane (GBM), the mesangial matrix (MM), the basement membrane of Bowman\u27s capsule (BBM) and the tubular basement membrane (TBM). JK132 only reacted with the MM, BBM and TBM. In Group B, JK199 reacted with GBM, MM, BBM, TBM and the interstitium (INS). JK132 only reacted with MM, BBM and TBM. In Group C and Group D, JK199 and JK132 reacted universally with GBM and INS in addition to MM, BBM and TBM. The intensity of the reaction was higher in Group D than in Group C. Thus, the reactivity of JK132 with GBM and INS was a unique finding for the CAN specimens. These results suggest that collagen IV is upregulated in CAN and the reactivity of JK132 in GBM and INS may represent the point of irreversible dysfunction of renal allografts