Urinary Tissue Inhibitor of Metalloproteinase and Insulin-like Growth Factor-7 as Early Biomarkers of Delayed Graft Function After Kidney Transplantation.

BACKGROUND: Recently, urinary tissue inhibitor of metalloproteinase-2 (TIMP-2) and insulin-like growth factor-7 (IGFBP-7), markers for G1 cell cycle arrest, have been identified and validated in predicting the development of acute kidney injury in critically ill patients. It is unknown, however, whether these two biomarkers could predict the development of delayed graft function (DGF) after kidney transplantation (KT). METHODS: This is a single-center, prospective, observational study. We enrolled 74 patients who underwent KT between August 2013 and December 2016. Urine sample were collected immediately after the operation. The primary outcome was development of DGF as defined by need for dialysis of more than 1 session within 7 days of KT. RESULTS: Twenty-three patients (31%) were diagnosed with DGF. In univariate analysis, kidneys from expanded criteria donors, higher donor serum creatinine, lower donor estimated glomerular filtration rate, antithymoglobulin exposure, neutrophil gelatinase associated lipocalin, and urinary [TIMP-2]·[IGFBP7] were significantly different between early graft function and DGF. However, in multivariate analysis adjusting other factors, deceased donor and urinary [TIMP-2]·[IGFBP7] at 0 hours post-transplantation could predict the development of DGF. The receiver operating characteristic curve for prediction of DGF showed an area under the curve of 0.867 (sensitivity 0.86, specificity 0.71) for a cutoff value of 1.39. CONCLUSIONS: Our results indicate that urine [TIMP-2]·[IGFBP7] immediately after transplantation could be an early, predictive biomarker of DGF in kidney transplantation.

Clinical outcomes in kidney transplantation patients from deceased donors with acute kidney injury.

BACKGROUND: This single-center study sought to examine the clinical outcomes of kidney transplant recipients from donors displaying acute kidney injury (AKI). METHODS: We analyzed retrospectively the medical records of the donors and recipients of 54 deceased-donor kidney transplantations performed in our center between March 2009 and March 2012. RESULTS: Among the 54 deceased donors, 36 (66.7%) experienced AKI as determined by the final mean serum creatinine levels measured before graft harvest of 2.66 ± 1.62 mg/dL versus 0.82 ± 0.28 mg/dL among non-AKI donors. The risks of delayed graft function and slow graft function were increased among the AKI versus non-AKI groups in the early post-transplantation period. However, the renal function status of recipients at 3, 6, and 12 months after transplantation was not significantly different between the two groups. Moreover, rejection-free survival rates during the study period were similar. Multivariate analysis revealed an acute rejection episodes (P = .047) and a lower body mass index in the donor relative to the recipient (P = .011) to be independent risk factors predicting poor graft function defined as a 1-year estimated glomerular filtration rate less than 50 mL/min/l.73 m(2). Donor AKI with either a high level (>4.0 mg/dL), an increasing trend of creatinine, or greater severity by the Risk, Injury, Failure, Loss, and End-stage kidney disease (RIFLE) classification was not a significant risk factor. CONCLUSION: Transplantation of kidneys from the AKI donors, namely, patients with severely decreased renal function, displayed excellent short-term outcomes. Accordingly, kidney transplantations from deceased donors with AKI should be considered more actively to expand the donor pool in Korea.

Urine neutrophil gelatinase-associated lipocalin predicts graft outcome up to 1 year after kidney transplantation.

BACKGROUND: Several recent reports demonstrated the usefulness of new biomarkers in early prediction of delayed graft function (DGF) and graft recovery after kidney transplantation (KT). It is unknown, however, whether these biomarkers would predict long-term graft outcome. In this study, we examined whether the biomarkers including neutrophil gelatinase-associated lipocalin (NGAL) and liver-type fatty acid-binding protein (L-FABP) can predict 1-year graft outcome as well as short-term graft function especially in patients with early graft function (EGF). METHODS: This was a single-center, prospective observational study. Urine samples at 0 hours and 2 and 6 days were obtained and the level of NGAL and L-FABP were measured. RESULTS: Of the 69 KT recipients enrolled, seven developed DGF, and the remaining 62 patients were finally enrolled as EGF recipients. EGF recipients were additionally divided into immediate graft function (IGF, n = 48) and slow graft function (SGF, n = 14) groups. Urinary NGAL (u-NGAL) level on day 2, but not L-FABP nor serum creatinine, was significantly higher in SGF compared to IGF group. Higher day 2 u-NGAL level was associated with more frequent development of SGF and, in addition, with significantly lower 1-year estimated glomerular filtration rate (eGFR). In multivariate logistic regression analysis, day 2 u-NGAL was a significant, independent factor for predicting poor long-term graft function (1-year eGFR < 60 mL/min/1.73 m(2)). CONCLUSIONS: This study demonstrates the possibility that u-NGAL might be useful in predicting adverse 1-year outcome as well as short-term graft function even in EGF patients.

Modification of radiation response in mice by Panax ginseng and diethyldithiocarbamate.

The aim of this study was to determine the effect of Panax ginseng and its fractions on jejunal crypt survival, endogenous spleen colony formation and apoptosis in jejunal crypt cells of mice irradiated with high- and low-dose of gamma-irradiation. The radioprotective effect of ginseng was compared with the effect of diethyldithiocarbamate (DDC). Ginseng administration before irradiation protected the jejunal crypts (p < 0.005), increased the formation of endogenous spleen colony (p < 0.005) and reduced the frequency of radiation-induced apoptosis (p < 0.05). The radioprotective effect on jejunal crypts and apoptosis in the DDC-treated group appeared similar to that in the ginseng--treated groups. Treatment with DDC showed no significant modifying effects on the formation of endogenous spleen colony. In the experiment on the effect of fractions of ginseng, the result indicated that the lipophilic non-polar compounds (Fraction 1), lipophilic-acidic compounds (Fraction 2), free sugars (Fraction 7) and saponin compounds (Fraction 8) might have a major radioprotective effect. Although the mechanisms of this inhibitory effect remain to be elucidated, these results indicated that ginseng might be a useful radioprotector, especially since it is a relatively nontoxic natural product. Further studies are needed to fully characterize the protective nature of ginseng extract and its components.

alpha-Melanocyte stimulating hormone (MSH) decreases cyclosporine a induced apoptosis in cultured human proximal tubular cells.

The pathogenesis of chronic cyclosporine A (CsA) nephrotoxicity has not been elucidated, but apoptosis is thought to play an important role in CsA induced tubular atrophy. Recently Fas-Fas ligand system mediated apoptosis has been frequently reported in many epithelial cells as well as in T lymphocytes. We investigated the ability of CsA to induce apoptosis in cultured human proximal tubular epithelial cells and also the effect of alpha-MSH on them. Fas, Fas ligand, and an intracellular adaptor protein, Fas-associating protein with death domain (FADD) expression, and poly-ADP ribose polymerase (PARP) cleavage were also studied. CsA induced apoptosis in cultured tubular epithelial cells demonstrated by increased number of TUNEL positive cells and it was accompanied by a significant increase in Fas mRNA and Fas ligand protein expressions. FADD and the cleavage product of PARP also increased, indicating the activation of caspase. In alpha-MSH co-treated cells, apoptosis markedly decreased with downregulation of Fas, Fas ligand and FADD expressions and also the cleavage product of PARP. In conclusion, these data suggest that tubular cell apoptosis mediated by Fas system may play a role in tubular atrophy in chronic CsA nephrotoxicity and pretreatment of alpha-MSH may have a some inhibitory effect on CsA induced tubular cell apoptosis.

A case of Rathke's Cleft Cyst inflammation presenting with diabetes insipidus.

Rathke's Cleft Cyst (RCC), which is located at the intrasellar region, is considered to be the distended remnants of Rathke's pouch, an invagination of the stomodeum. Lined with columnar or cuboidal epithelium of ectodermal origin, RCC usually contains mucoid material and it is found in 13-22% of normal pituitary glands. The cyst rarely leads to the development of symptoms but, when it does, the most common presenting symptoms are headache, visual impairment, hypopituitarism and hypothalamic dysfunction. However, in some cases it presents symptoms of diabetes insipidus, decreased libido and impotence. Recently we experienced a case of RCC inflammation presenting with diabetes insipidus and treated with transsphenoidal surgery. To our knowledge, this is the first report of RCC presenting with symptoms of diabetes insipidus in Korea.

alpha-MSH decreases apoptosis in ischaemic acute renal failure in rats: possible mechanism of this beneficial effect.

BACKGROUND: Apoptosis frequently occurs in acute renal injury but the molecular mechanisms responsible for this distinct form of cell death are largely unknown. Fas belongs to the tumour necrosis factor (TNF)/nerve growth factor superfamily and engagement by Fas ligand induces apoptosis in various epithelial cells. To investigate the role of apoptosis and associated mechanisms, we examined the occurrence of apoptosis and Fas and Fas ligand expression, and the therapeutic effect of alpha-melanocyte-stimulating hormone (alpha-MSH), a potent anti-inflammatory cytokine in an ischaemic acute renal failure (ARF) rat model. We also examined neutrophil infiltration together with intercellular adhesion molecule-1 (ICAM-1) expression because of their possible involvement in apoptosis due to their ability to release various inflammatory cytokines and reactive oxygen species. METHODS: After unilateral nephrectomy in female Sprague-Dawley rats, the renal artery of the contralateral kidney was clamped for 40 min and reperfused. alpha-MSH or vehicle was injected intraperitoneally immediately after reperfusion and at 1, 6, or 24 h after reperfusion. The expression of Fas and Fas ligand was studied by western blot analysis and semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) method, and neutrophil infiltration by naphthol AS-D chloracetate staining. The degree of apoptosis, neutrophil infiltration, and Fas and Fas ligand, and ICAM-1 expression, as well as biochemical and histological data were compared between the alpha-MSH and the vehicle-treated groups. RESULTS: Intraperitoneally administered alpha-MSH significantly reduced renal injury, measured by blood urea nitrogen (BUN) and creatinine and by the degree of tubular necrosis (109.6+/-7.1/54.7+/-3.1 mg/dl for BUN, and 1.6+/-0.2/1.03+/-0.06 mg/dl for creatinine 24 h after ischaemia) (5.4+/-0.8/2.6+/-0.3 for injury score 24 h after ischaemia). Ischaemia caused an increase in Fas and Fas ligand expression and was accompanied by morphological evidence of apoptosis. alpha-MSH significantly reduced the degree of apoptosis, as well as Fas and Fas ligand expression (mean apoptotic cell number, 41.7+/-3.5/14.2+/-2.2 per x200 field at 24 h after ischaemia. Fas protein expression: sham, 1409+/-159 DI (densitometric index); vehicle/alpha-MSH, 2818+/-635/1306+/-321 DI at 24 h and 5542+/-799/2867+/-455 DI at 72 h after ischaemia. Fas ligand protein expression: sham, 1221+/-181 DI; vehicle/alpha-MSH, 2590+/-85/1279+/-169 DI at 4 h, 4376+/-268/2432+/-369 DI at 24 h and 5200+/-648/2253.7+/-1104 DI at 72 h after ischaemia). Neutrophil infiltration and ICAM-1 expression were also significantly reduced in alpha-MSH group (neutrophil infiltration: vehicle/ alpha-MSH, 5.05+/-1.8/1.59+/-0.4) (ICAM-1 expression, vehicle/alpha-MSH 0.46+/-0.21/0.29+/-0.19). CONCLUSION: These results suggest that apoptosis clearly contributes to tubular cell loss in ischaemia/reperfusion (I/R) injury possibly by neutrophil-mediated pathways or an increase in Fas-Fas ligand expression. The observed beneficial effect of alpha-MSH could be related to these mechanisms.

Influence of gestational age at exposure on the prenatal effects of gamma-radiation.

The objective of this investigation was to evaluate the influence of gestational age at exposure on the prenatal effects of gamma-radiation. Pregnant ICR mice were exposed to a single dose of 2.0 Gy gamma-radiation at a gestational 2.5 to 15.5 days post-coitus (p.c.). The animals were sacrificed on day 18 of gestation and the fetuses were examined for mortality, growth retardation, change in head size and any other morphological abnormalities. The only demonstrable effect of irradiation during the preimplantation period was an increase in prenatal mortality. Resorptions were maximal on post-exposure day 2.5 after conception. The pre-implantation irradiated embryos which survived did not show any major fetal abnormalities. Small head, growth retardation, cleft palate, dilatation of the cerebral ventricle, dilatation of the renal pelvis and abnormalities of the extremities and tail were prominent after exposure during the organogenesis period, especially on day 11.5 of gestation. Our results indicate that the late period of organogenesis in the mouse is a particularly sensitive phase in terms of the development of the brain, skull and extremities.

Dependence of malformation upon gestational age and exposed dose of gamma radiation.

In order to evaluate the importance of gestational age and the dose-incidence relationship by gamma radiation, pregnant ICR mice at gestational days from 2.5 to 15.5 days post-coitus (p.c.) were exposed to a single dose of 2.0 Gy and also at day 11.5 after conception, which was the most sensitive stage for the induction of major congenital malformations. The animals were sacrificed on day 18 of gestation and the fetuses were examined for mortality, growth retardation, changes in head size and other morphological abnormalities. The only demonstrable effect of irradiation during the pre-implantation period was an increase in prenatal mortality. Resorptions were maximal on exposure at day 2.5 after conception. The pre-implantation irradiated embryos which survived did not show any major fetal abnormalities. A small head, growth retardation, a cleft palate, dilatation of the cerebral ventricle, a renal pelvis, and abnormalities of the extremities and tail after exposure were prominent during the organogenesis period, especially on day 11.5 of gestation. As for the dose-incidence relationship, the incidence of a small head, growth-retarded fetuses, a cleft palate, dilatation of cerebral ventricle and abnormalities of the extremities in live fetuses rose as the radiation dose increased. The result indicated that the late period of organogenesis in the development of the brain, skull and extremities of a mouse was a particularly sensitive phase. The threshold doses of radiation that induced a cleft palate and dilatation of the cerebral ventricle, and abnormal extremities were between 1.0 and 2.0 Gy, and between 0.5 and 1.0 Gy, respectively.

Role of vascular endothelial growth factor in diabetic nephropathy.

BACKGROUND: Vascular endothelial growth factor (VEGF) is a potent cytokine that is considered to be an important mediator in the pathogenesis of endothelial dysfunction in diabetes. METHODS: This study investigates the effect of high glucose on the signaling and production of VEGF in rat mesangial cells in culture and measures the urinary VEGF level in patients with different stages of diabetic nephropathy. To elucidate the role of VEGF in vivo further, expression of VEGF in control and diabetic kidneys was examined using immunohistochemistry. RESULTS: A high ambient glucose concentration in the culture medium increased VEGF mRNA expression and protein production within 3 h in a concentration-dependent manner. A protein kinase C (PKC) inhibitor and PKC down-regulation inhibited glucose-induced increases in VEGF production. Urinary excretion of VEGF significantly increased according to the degree of proteinuria in patients with diabetes. A weak but significant correlation was found between urinary VEGF excretion and the levels of serum creatinine, creatinine clearance, microalbuminuria, and proteinuria. Immunohistochemistry revealed marked differences in the extent of mesangial VEGF staining between diabetic and control kidneys. Pronounced up-regulation of VEGF was observed in the glomerular epithelial cell in the early phase of diabetic kidney disease, whereas widespread expression of VEGF was found in the tubular segments, especially the proximal segment, in advanced diabetic nephropathy. CONCLUSIONS: These results suggest that VEGF may play a role in the pathogenesis of diabetic nephropathy.

Establishment of a biological indicator for the radiation and safety of diagnostic ultrasound using apoptosis.

We have studied, by a nonisotopic in situ end-labeling (ISEL) technique, the frequency of apoptosis in the intestinal crypt cell of adult mice and in the external granular layer(EGL) of the cerebellum of fetuses by gamma-ray irradiation from 60Co or diagnostic ultrasound exposure. The extent of changes following 200 cGy(1090 cGy/min) was studied at 2, 4, 6, 8, 12, or 24 hours after exposure. The maximal frequency was found 4-8 hours after exposure. The mice that received 18, 36, 54, 108, 198, 396 cGy of gamma-rays or diagnostic ultrasound (7.5 MHz, 4.2 mW, ISPTA = 7.9 mW/cm2, IsppA = 114.3 W/cm2) for 10 or 30 minutes were examined 6 hours after irradiation. Measurements performed after gamma-ray irradiation showed a dose-related increase in apoptotic cells in each of the mice studied. The dose-response curves were analyzed with alpha linear-quadratic model: the frequency (number per crypt) of apoptotic cells in the intestinal crypt of adult mice was y = (0.0386 +/- 0.004204)D + (-0.0000535 +/- 0.00001120)D2 + 0.15475(r2 = 0.952, y = apoptotic cell per crypt cell, D = dose in cGy), and the frequency (percentage of apoptotic cell in the EGL) of apoptotic cell in the EGL of fetus was y = (0.1349 +/- 1.175)D + (-0.001522 +/- 0.334)D2 + 0.0477(r2 = 0.981, y = % of apoptotic cell in the EGL, D = dose in cGy). In the experiment of ultrasound exposure, the frequencies of apoptosis were 0.181 +/- 0.055(10 minutes exposure) and 0.325 +/- 0.294 (30 minutes exposure) in the crypt cells and 0.106 +/- 0.130% (10 minutes exposure) and 0.167 +/- 0.220%(30 minutes exposure) in the EGL. We estimated the relative dose of the yield from the experiment with ultrasound by substituting the yield from ultrasound exposure into the curue from the gamma-irradiation. The relative doses of ultrasound exposure compared with gamma-irradiation were 0.692 cGy(10 minutes exposure) and 1.334 cGy(30 minutes exposure) in the experiment for crypt cells and 0.432 cGy(10 minutes exposure) and 0.885 cGy(30 minutes exposure) in the experiment for EGL. Although there is presently no evidence to indicate that diagnostic ultrasound involves a significant risk, it is not wise to use diagnostic ultrasound indiscriminately.

ACE gene polymorphism and renal responsiveness to ACE inhibitors in IgA nephropathy patients.

We examined the renal responsiveness to ACE inhibitor in IgA nephropathy (IgAN) patients according to the grouping of ACE gene polymorphism. Sixty one patients diagnosed as IgAN by renal biopsy and prescribed with ACE inhibitors were enrolled. Genomic DNA was extracted from whole blood and PCR was performed. The I/D polymorphism was determined by the presence of the 287 bp fragment in intron 16 of chromosome 17. During the follow-up period (mean; 44.6 months, median: 44.5 months, 5 to 113 months), the blood pressure of 61 patients was controlled below 130/80 mmHg. The renal responsiveness was determined by the degree of changes of proteinuria at 12 months after initiation of ACE inhibitors and by the slope of reciprocal variation of the serum creatinine against follow-up duration ¿(1/Cr2-1/Cr1)/durations¿. The distribution of the II, ID and DD genotype among 61 patients was 21, 16 and 24 patients, respectively. There were no differences among three genotypes in age, sex, the number of patients with initial blood pressure over 140/90 mmHg, initial serum creatinine level, the number of patients with initial azotemia (> 1.4 mg/dL) and with initial 24-hr proteinuria amount over 2.0 g. Significant anti-proteinuric effect of ACE inhibitor was found in IgAN (p = 0.001), but no significant difference was found among genotypes. Significant difference (p = 0.011) was noticed between II type and DD type in the slope of reciprocal variation of the serum creatinine against follow-up duration. In conclusion, efficacy of ACE inhibitors on renal function preservation in IgAN was more pronounced in DD genotype than II genotype.

A case of Wegener's granulomatosis complicated by diffuse pulmonary hemorrhage and thrombotic thrombocytopenic purpura.

Wegener's granulomatosis is a distinct form of necrotizing granulomatous vasculitis which usually affects the kidneys and the upper and lower respiratory tracts. Unusual manifestations have also been reported, and these include colitis, urethritis and diabetes insipidus. We describe a case of Wegener's granulomatosis which presented with rapidly progressive renal insufficiency, sudden deafness, red eye, facial palsy, and complicated by uncommon manifestations that were diffuse pulmonary hemorrhage and thrombotic thrombocytopenic purpura.

Induction of micronuclei in human, goat, rabbit peripheral blood lymphocytes and mouse splenic lymphocytes irradiated in vitro with gamma radiation.

The frequencies of gamma-ray-induced micronuclei (MN) in cytokinesis-blocked (CB) lymphocytes at several doses were measured in three donors of four species (human, goat, rabbit, mouse). Measurements performed after irradiation showed a dose-related increases in MN frequency in each of the donors studied. The relative sensitivity of mouse in spleen lymphocytes (SLs), goat in peripheral blood lymphocytes (PBLs) and rabbit PBLs compared with human PBLs was estimated by best fitting linear-quadratic model based on the radiation-induced MN data over the range from 0 to 400 cGy. In the case of MN frequency with 0.2, the relative sensitivities of mouse SLs, goat PBLs and rabbit PBLs were 1.67, 0.98 and 0.39, respectively. These data indicate that the induction of MN in CB cells following irradiation is similar in human and goat PBLs, and PBLs from rabbit were much less sensitive to the MN induction effects of gamma-radiation than those from human. Compared with the radiation-induced MN formation in the PBLs of human, the SLs of mouse were more radiosensitive.

Inhibition of autochthonous tumor by ethanol insoluble fraction from Panax ginseng as an immunomodulator.

The water extract of Panax ginseng was fractionated by its solubility in ethanol and then the ethanol-insoluble fraction was tested for immunomodulatory activity. The ethanol-insoluble fraction of ginseng (Fr. 3) proliferated splenocytes and generated activated killer cells in vitro. These activated killer cells killed both NK cell sensitive and insensitive tumor target cells without MHC-restriction. Activation of splenocytes by ginseng was mediated through the endogenously produced IL-2. To investigate the effects of Fr.3 on the autochthonous neoplasm, a single subcutaneous injection of 0.5 mg of benzo[a]pyrene (BP) was given within 24 hours after birth of male N: GP(S) mice, and Fr.3 was administered in drinking water at a concentration of 2 mg/ml, 1 mg/ml, or 0.5 mg/ml for 6 weeks after weaning. The treatment with Fr. 3 significantly inhibited lung tumor incidence (P < 0.05) compared with the BP alone group at a concentration of 2 mg/ml or 1 mg/ml in drinking water at the 9th week after BP treatment. These results suggest that the ethanol-insoluble fraction of ginseng shows antitumor effects as an immunomodulator.

Effect of red ginseng on natural killer cell activity in mice with lung adenoma induced by urethan and benzo(a)pyrene.

It was previously reported that red ginseng extract inhibited carcinogenesis by urethan, DMBA, and aflatoxin B1 [Yun et al: Cancer Detect Prevent 1983; 6:515-25]. In an attempt to investigate the mechanism of the anticarcinogenic effect of ginseng, the natural killer (NK) activity and the incidence of lung adenoma were followed over a period of 48 weeks postinjection with urethan or benzo(a)pyrene. The NK activity was markedly depressed from 4 weeks to 24 weeks after injection of carcinogens. This decreased NK activity was returned to the level of controls by administration of ginseng. At the same time, a lower incidence of lung adenoma was noted following administration of ginseng to urethan-injected mice. However, the lung adenoma induced by benzo(a)pyrene began to occur at 48 weeks in which NK activity had naturally declined to a level too low to be affected by ginseng, and administration of ginseng did not decrease the incidence. In conclusion, these results suggest that the anticarcinogenic effect of ginseng may be related to the augmentation of NK activity.