PURPOSE: CAPD is an important treatment modality along with hemodialysis and kidney transplantation in end stage renal disease. Malnutrition is very common and associated with increased morbidity and mortality in CAPD patients. The cause of malnutrion in CAPD patients might be multifactorial. This prospective study was carried out to investigate nutritional changes for 1 year after initiation of peritoneal dialysis by measurement body composition, especially lean body mass (LBM) using bioelectrical impedance analysis (BIA) and dual energy X-ray absorptiometry (DEXA) and to evaluate the factors associated with malnutrition in CAPD patients. METHODS: Among new CAPD patients from May, 2001 to Dec, 2002 in our hospital, 25 patients were enrolled. Body weight, LBM, LBM percen t (%LBM), fat mass, fat mass percent (%fat mass), ECF volume and ECF/TBW were compared between 1st month and 12th month after initiation of PD. The biochemical parameters, Urea kinetic modeling, Peritoneal equilibration test, the amounts of glucose absorption through the dialysate, the amounts of protein and albumin loss through the dialysate were measured at the same time point with measurement of the body composition. RESULTS: There were significantly decreased LBM (46.3+/-9.1 kg to 44.7+/-9.0 kg in BIA, 45.7+/-9.3 kg to 42.1+/-7.9 kg in DEXA, p< 0.05, respectively) but significantly increased fat mass (16.3+/-6.2 kg to 20.2+/-7.9 kg in BIA, 15.7+/-6.6 kg to 20.1+/-7.4 kg in DEXA, p<0.01, respectively) during first one year. Mean weekly Kt/V were significantly correlated with the changes of LBM (r=-0.64 in BIA, r=-0.81 in DEXA, p<0.01, respectively). With the multiple regression test, 1st month weekly Kt/V in BIA and DEXA were significant predictors of the changes of LBM for 1 year (beta-coefficients: -0.573 in BIA, -0.773 in DEXA, p<0.01, respectively). CONCLUSION: Adequate dialysis, especially 1st month adequacy, is very important for maintaining good nutritional status for one year after initiation of peritoneal dialysis.
Absorptiometry, Photon ; Absorption ; Body Composition* ; Body Weight ; Dialysis ; Electric Impedance ; Glucose ; Humans ; Kidney Failure, Chronic ; Kidney Transplantation ; Malnutrition ; Mortality ; Nutritional Status ; Peritoneal Dialysis ; Peritoneal Dialysis, Continuous Ambulatory* ; Prospective Studies ; Renal Dialysis ; Urea

Patients with cardiac rupture due to blunt trauma have more than 50% mortality rate and most of them expire before they arrive at the hospital emergency room. Since patients typically present with cardiac tamponade, the diagnosis can be easily confirmed with physical examination, echocardiography, and chest CT scan. However, in our case of the massive hemothorax on right side without evidence of cardiac tamponade, the diagnosis for cardiac rupture does not seems to be easy. Therefore, we must assume the probability of cardiac rupture if we plan an explo-thoracotomy in a patients with massive right hemothorax without rib fracture. We describe two cases of cardiac rupture combined with pericardial laceration and right massive hemothorax by blunt chest trauma. The ruptured hearts of the patients were successfully closed using cardio-pulmonary bypass or cell saver system without detrimental sequelae.
Cardiac Tamponade ; Diagnosis ; Echocardiography ; Emergency Service, Hospital ; Heart ; Heart Rupture* ; Hemothorax* ; Humans ; Lacerations ; Mortality ; Physical Examination ; Rib Fractures ; Thorax* ; Tomography, X-Ray Computed

PURPOSE: Islet cell transplantation, as an alternative approach to endocrine cell replacement to treat the diabetes mellitus, has received significant attention because it holds several advantages over whole gland transplantation. However cell damage from islet isolation and immunologic rejection after transplantation prevent from successful clinical application for diabetic patients. Culture of cells at low temperature has known to stabilize the cell viability, and to decrease the immunologic antigenicity. Aim of this study is to investigate the effect of culture at 24oC on cell viability, cellular function, immunogenicity and cytokine profiles in rat pancreas islet. METHODS: Pancreas islets were isolated from Lewis rat and cultured at 24oC or 37oC during 14 days. Islet recovery after culture period was counted as islet equivalent number, and islet viability was examined with fluorescent vital staining (FDA/PI). Islet function was measured with glucose stimulation test. Annexin V expression and MHC class I and II expression were measured with flow cytometric assay for apoptosis and immunogenicity respectively. Lymphocyte cell proliferation through mixed lymphocyte islet culture was examined with WST-1 proliferation assay. Cytokine profiles were analyzed with quantitative real time RT-PCR. All these parameters were measured on 1, 3, 5, 7, 14 culture days after islet isolation. RESULTS: Islet recovery was higher in islet cultured at 24oC than in islet cultured at 37oC without change of viability. Insulin secretion after glucose stimulation was more effective in 24oC culture condition. Decrease of apoptotic cell death was demonstrated in 24oC cultured islet. MHC class I and II expression on islets and lymphocyte proliferation when cocultured with islets were less prominent in 24oC cultured islet. TNF-alpha and IL-4 cytokine expression was higher in islet cultured at 24oC than in islet cultured at 37oC. IL-1beta and IL-10 cytokine expression were similar in both culture condition. CONCLUSION: This study demonstrated that cell recovery and function are increased in islet cultured at 24oC than in islet cultured at 37oC while antigenicity and proinflammatory cytokine expression are decreased. Low temperature culture can be a good approach to prevent the loss of islet mass, and to reduce the immunologic rejection of transplanted islet for successful clinical islet transplantation.
Animals ; Annexin A5 ; Apoptosis ; Cell Death ; Cell Proliferation ; Cell Survival ; Diabetes Mellitus ; Endocrine Cells ; Glucose ; Humans ; Insulin ; Interleukin-10 ; Interleukin-4 ; Islets of Langerhans Transplantation ; Islets of Langerhans* ; Lymphocytes ; Pancreas* ; Rats* ; Tumor Necrosis Factor-alpha

Ceramide metabolism is known to be an essential etiology for various diseases, such as atopic dermatitis and Gaucher disease. Glucosylceramide synthase (GCS) is a key enzyme for the synthesis of glucosylceramide (GlcCer), which is a main ceramide metabolism pathway in mammalian cells. In this article, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine GCS activity using synthetic non-natural sphingolipid C8-ceramide as a substrate. The reaction products, C8-GlcCer for GCS, could be separated on a C18 column by reverse-phase high-performance liquid chromatography (HPLC). Quantification was conducted using the multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z 588.6 → 264.4 for C8-GlcCer at positive ionization mode. The calibration curve was established over the range of 0.625–160 ng/mL, and the correlation coefficient was larger than 0.999. This method was successfully applied to detect GCS in the human hepatocellular carcinoma cell line (HepG2 cells) and mouse peripheral blood mononuclear cells. We also evaluated the inhibition degree of a known GCS inhibitor 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) on GCS enzymatic activity and proved that this method could be successfully applied to GCS inhibitor screening of preventive and therapeutic drugs for ceramide metabolism diseases, such as atopic dermatitis and Gaucher disease.
Animals ; Calibration ; Carcinoma, Hepatocellular ; Cell Line ; Chromatography, Liquid ; Dermatitis, Atopic ; Gaucher Disease ; Humans ; Mass Screening ; Mass Spectrometry ; Metabolism ; Methods ; Mice

A 60-year-old woman with end stage liver cirrhosis caused by genotype 2 hepatitis C virus (HCV) infection received an orthotopic liver transplantation (OLT). The patient was negative for the hepatitis B surface antigen (HBsAg) and positive for the anti-hepatitis B surface antibody (anti-HBs) prior to and one and a half months following the OLT. Due to reactivation of hepatitis C, treatment with interferon-alpha and Ribavirin started two months following the OLT and resulted in a sustained virological response. We performed a liver biopsy because a biochemical response was not achieved. Surprisingly, liver pathology showed HBsAg-positive hepatocytes with a lobular hepatitis feature, which had been negative in the liver biopsy specimen obtained one and a half months post-OLT. High titers of both HBsAg and HBeAg were detected, while anti-HBs antibodies were not found. Tests for IgM anti-hepatitis B core antibody and anti-delta virus antibodies were negative. The serum HBV DNA titer was over 1x10(7) copies/mL. A sequencing analysis showed no mutation in the "a" determinant region, but revealed a mixture of wild and mutant strains at an overlapping region of the S and P genes (S codon 213 (Leu/Ile); P codons 221 (Phe/Tyr) and 222 (Ala/Thr)). These findings suggest that de novo hepatitis B can develop in patients with HCV infection during the post-OLT period despite the presence of protective anti-HBs.
Antibodies ; Biopsy ; Codon ; DNA ; Female ; Genotype ; Hepacivirus ; Hepatitis ; Hepatitis B ; Hepatitis B e Antigens ; Hepatitis B Surface Antigens ; Hepatitis B virus ; Hepatitis C ; Hepatocytes ; Humans ; Immunoglobulin M ; Interferon-alpha ; Liver ; Liver Cirrhosis ; Liver Transplantation ; Middle Aged ; Ribavirin ; Superinfection ; Viruses

Paraquat dichloride (N,N-dimethyl-4-4'-bipiridinium, PQ) is an extremely toxic chemical that is widely used in herbicides. PQ generates reactive oxygen species (ROS) and causes multiple organ failure. In particular, PQ has been reported to be an immunotoxic agrochemical compound. PQ was shown to decrease the number of macrophages in rats and suppress monocyte phagocytic activity in mice. However, the effect of PQ on macrophage cell viability remains unclear. In this study, we evaluated the cytotoxic effect of PQ on the mouse macrophage cell line, RAW264.7 and its possible mechanism of action. RAW264.7 cells were treated with PQ (0, 75, and 150 muM), and cellular apoptosis, mitochondrial membrane potential (MMP), and intracellular ROS levels were determined. Morphological changes to the cell nucleus and cellular apoptosis were also evaluated by DAPI and Annexin V staining, respectively. In this study, PQ induced apoptotic cell death by dose-dependently decreasing MMP. Additionally, PQ increased the cleaved form of caspase-3, an apoptotic marker. In conclusion, PQ induces apoptosis in RAW264.7 cells through a ROS-mediated mitochondrial pathway. Thus, our study improves our knowledge of PQ-induced toxicity, and may give us a greater understanding of how PQ affects the immune system.
Animals ; Annexin A5 ; Apoptosis* ; Caspase 3 ; Cell Death ; Cell Line ; Cell Nucleus ; Cell Survival ; Herbicides ; Immune System ; Macrophages ; Membrane Potential, Mitochondrial ; Mice ; Mitochondria ; Monocytes ; Multiple Organ Failure ; Paraquat* ; Rats ; Reactive Oxygen Species

PURPOSE: Cryopreservation of pancreas islet cells can facilitate the clinical islet transplantation by giving a means of storage of islets and immunomodulation on pancreatic islet preparations. METHODS: Pancreatic islets were isolated by standard technique using collagenase in rat. Cryopreservation was performed by using DMSO as a cryoprotectant after one day or 48 hr culture. Recovery rate, viability and insulin release in assay in vitro and vivo were checked under the various conditions, such as concentration of DMSO (1.5 M or 2.0 M), culture condition (1 day or 2 day), and taurine treatment. RESULTS: Percentage of recovery of cryopreserved islet was 56.8+/-10% after thawing. Viability was decreased from 97.2+/-1.1% before cryopreservation to 82.7+/-9.9% after thawing. Glucose stimulation index was reduced from 1.7+/-0.2 before cryopreservation to 1.2+/-0.8 after thawing. Nucleation method by metal rod showed better viability than control (no nucleation) or chamber nucleation method. Mean viability and glucose stimulation index was 81% and 1.5 in one day culture, and 84.2% and 1.2 in 2 day culture before cryopreservtion. Islet treated with taurine showed better insulin release and intracellular insulin content compared with non treated islets before and after cryopreservation. When 4000 IEQ (Islet Equivalent) of islets treated with taurine and non treated cryopreserved islets were transplanted into syngenic streptozotocin induced diabetic rat, all showed normoglycemia over 60 days. CONCLUSION: Cryopreservation of islets could give a tool of storage with preservation of islet secretion function. However pertinent effort to improve the recovery is needed in order to be used in the clinical islet transplantation.
Animals ; Collagenases ; Cryopreservation* ; Dimethyl Sulfoxide ; Glucose ; Immunomodulation ; Insulin ; Islets of Langerhans Transplantation ; Islets of Langerhans* ; Pancreas* ; Rats* ; Streptozocin ; Taurine

Rheumatoid arthritis (RA) is a multifactorial immune-mediated disease, the pathogenesis of which involves different cell types. T-cell activation plays an important role in RA. Therefore, inhibiting T-cell activation is one of the current therapeutic strategies.Cytotoxic T-lymphocyte antigen 4-immunoglobulin (CTLA4-Ig), also known as abatacept, reduces cytokine secretion by inhibiting T-cell activation. To achieve a homeostatic therapeutic effect, CTLA4-Ig has to be administered repeatedly over several weeks, which limits its applicability in RA treatment. To overcome this limitation, we increased the number of sialic acid-capped antennas by genetically engineering the CTLA4 region to increase the therapeutic effect of CTLA4-Ig. N-acetylglucosaminyltransferase (GnT) and α2,6-sialyltransferase (α2,6-ST) were co-overexpressed in Chinese hamster ovary (CHO) cells to generate a highly sialylated CTLA4-Ig fusion protein, named ST6. The therapeutic and immunogenic effects of ST6 and CTLA4-Ig were compared. ST6 dose-dependently decreased paw edema in a mouse model of collagen-induced arthritis and reduced cytokine levels in a co-culture cell assay in a similar manner to CTLA4-Ig. ST6- and CTLA4-Ig-induced T cell-derived cytokines were examined in CD4 T cells isolated from peripheral blood mononuclear cells after cell killing through irradiation followed by flow- and magnetic-beadassisted separation. Interestingly, compared to CTLA4-Ig, ST6 was substantially less immunogenic and more stable and durable. Our data suggest that ST6 can serve as a novel, less immunogenic therapeutic strategy for patients with RA.