INSULIN AND MINERALOCORTICOIDS INFLUENCE ON EXTRARENAL POTASSIUM METABOLISM IN CHRONIC HEMODIALYSIS PATIENTS

Insulin-mineral corticoids effects on extrarenal K⁺ metabolism in dialysis patients. During the inter-dialytic interval in dialyzed patients, hydrogen and potassium ions are regulated by extrarenal mechanisms. We studied the hormonal and acidotic effects on the extrarenal potassium metabolism, in selected, anuric and stable, hemodialysis patients. Fifteen patients, were grouped according to the mean mid-week pre-dialysis K⁺ over the past 12 months: > 6.0 mEq/L (G1, n = 5), = 5.1–6.0 mEq/L (G2, n = 5), ≤5.0 mEq/L (G3, n = 5). After a mid-week hemodialysis session and 12 h fasting, they received 1 g/Kg glucose p.os (A). Insulin, aldosterone, renin, pH, HCO₃^?, glucose, body weight, blood pressure and heart rate were measured before and 60′ after the meal. We recorded the same parameters, except insulin, in 15 patients, similarly grouped, before hemodialysis (T0) and on 3 consecutive off dialysis days (T1–T3); G1 received fluorohydrocortisone (FHC) 0.1 mg–0.3 mg/day, according to body weight and G3 spironolactone (SLT) 200 mg per day. G2 were controls (B). (A) A significant rise in glycemia (81 ± 23 to 157 ± 52 mg/dL, P < 0.001) and insulin (11.8 ± 6.2 to 46.8 ± 19.5 μU/mL, P<0.001), with a drop in K⁺ (5.1 ± 0.6 to 4.8 ± 0.7 mEq/L, P= 0.001) and aldosterone (453 ± 373 to 383 ± 364 pg/mL, P<0.01), were noted at T60 vs. T0, in all groups. Insulin levels correlated negatively (r = ?0.54, P<0.04) to serum K⁺ at T60, in all patients. (B) No major pH, HCO₃ and aldosterone changes were observed in the 3 groups. Despite that, K⁺ dropped in G1 by FHC (6.7 ± 0.9 to 5.9 ± 0.6 mEq/L, P<0.05), rose in G3 by SLT (4.4 ± 0.4 to 5.4 ± 0.3 mEq/L, P<0.05) and remained unchanged in controls (5.8 ± 0.2 to 5.8 ± 0.6 mEq/L), (T0 vs T3 pre-dialysis values). Glucose significantly lowered K^? by promoting adequate insulin secretion. Drugs affecting aldosterone action significantly influenced potassium metabolism. Acid-base balance was not important in K⁺ handling in steady state anuric dialysis patients.     

DNA Oxidative Damage in Patients with Dialysis Treatment

Background/Aims. Chronic renal patients on hemodialysis (HD) and peritoneal dialysis (PD) treatment are exposed to oxidative stress and DNA damage. The objective of this study was to assess the oxidative damage to DNA in end-stage chronic renal failure, before and after vitamin E supplementation. Methods. Patients on HD (n = 29) and PD (n = 22) received oral supplementation with 300 mg vitamin E three times a week for 4 weeks. A blood sample was collected at the beginning and at the end of the supplementation cycle for the determination of vitamin E levels (high-performance liquid chromatography), carbonyl groups, and DNA damage (8-hydroxy 2′-deoxyguanosine [8-OHdG] and comet assay). Results. After supplementation, vitamin E concentration was increased by about 50%. Protein oxidation was initially observed in both groups, with a reduction after supplementation. DNA damage detected by the comet assay and by 8-OHdG analysis was significantly reduced (p< 0.05) after supplementation in both groups. Conclusions. Vitamin E supplementation reduced oxidative DNA damage in both HD and PD patients. Treatments such as HD and PD induce oxidative stress and consequent DNA damage, and increased plasma vitamin E levels significantly contribute to the normalization of these events.     

Evaluation of oxidative DNA damage in human sperm and its association with male infertility

Recently, there is increasing evidence suggesting that oxidative sperm DNA damage is closely associated with impaired sperm function and male infertility. 8-Hydroxydeoxyguanosine (8-OHdG) is considered to be a precise and sensitive biomarker of oxidative DNA damage. The present study was thus designed to evaluate the extent of oxidative DNA damage in sperm and its association with male infertility by assaying the 8-OHdG levels in human sperm samples. A total of 114 subjects (60 infertile patients and 54 age-matched healthy workers) participated in this study. The level of 8-OHdG in sperm DNA was determined by high-performance liquid chromatography with electrochemical detection, and the conventional seminal parameters were also measured according to World Health Organization guidelines. It was found that the level of sperm 8-OHdG in infertile patients was significantly higher than that in healthy subjects (10.03 vs. 4.79 8-OHdG/10(5) dG; geometric mean, P < 0.001). The correlation between sperm 8-OHdG levels and conventional seminal parameters were also analyzed. There is a significant positive correlation between 8-OHdG and sperm head defects (r = 0.38, P < 0.001), whereas significant inverse correlations were noted for 8-OHdG with sperm density (r = -0.42, P < 0.001), total sperm number (r = -0.42, P < 0.001), sperm motility (r = -0.24, P < 0.01), and normal sperm morphology (r = -0.39, P < 0.001). Data from this study thus indicate that oxidative damage to sperm DNA may be important in the etiology of male infertility and that the assay of sperm 8-OHdG may have potential diagnostic value in the evaluation of sperm function and male fertility.     

Cigarette Smoking and hOGG1 Ser326Cys Polymorphism are Associated with 8-OHdG Accumulation on Mitochondrial DNA in Thoracic Esophageal Squamous Cell Carcinoma

Background

We examined whether cigarette smoking affects the degrees of oxidative damage (8-hydroxyl-2′-deoxyguanosine [8-OHdG]) on mitochondrial DNA (mtDNA), whether the degree of 8-OHdG accumulation on mtDNA is related to the increased total mtDNA copy number, and whether human 8-oxoguanine DNA glycosylase 1 (hOGG1) Ser326Cys polymorphisms affect the degrees of 8-OHdG accumulation on mtDNA in thoracic esophageal squamous cell carcinoma (TESCC).

Methods

DNA extracted from microdissected tissues of paired noncancerous esophageal muscles, noncancerous esophageal mucosa, and cancerous TESCC nests (n = 74) along with metastatic lymph nodes (n = 38) of 74 TESCC patients was analyzed. Both the mtDNA copy number and mtDNA integrity were analyzed by quantitative real-time polymerase chain reaction (PCR). The hOGG1 Ser326Cys polymorphisms were identified by restriction fragment length polymorphism PCR and PCR-based direct sequencing.

Results

Among noncancerous esophageal mucosa, cancerous TESCC nests, and metastatic lymph nodes, the mtDNA integrity decreased (95.2 to 47.9 to 18.6 %; P < 0.001) and the mtDNA copy number disproportionally increased (0.163 to 0.204 to 0.207; P = 0.026). In TESCC, higher indexes of cigarette smoking (0, 0–20, 20–40, and >40 pack-years) were related to an advanced pathologic N category (P = 0.038), elevated mtDNA copy number (P = 0.013), higher mtDNA copy ratio (P = 0.028), and increased mtDNA integrity (P = 0.069). The TESCC mtDNA integrity in patients with Ser/Ser, Ser/Cys, and Cys/Cys hOGG1 variants decreased stepwise from 65.2 to 52.1 to 41.3 % (P = 0.051).

Conclusions

Elevated 8-OHdG accumulations on mtDNA in TESCC were observed. Such accumulations were associated with a compensatory increase in total mtDNA copy number, indexes of cigarette smoking, and hOGG1 Ser326Cys polymorphisms.

     

Effects of insulin and antioxidant on plasma 8-hydroxyguanine and tissue 8-hydroxydeoxyguanosine in streptozotocin-induced diabetic rats.

Cumulating evidence suggests that enhanced oxidative stress may contribute to diabetic angiopathy. The levels of 8-hydroxydeoxyguanosine (8-OHdG) and 8-hydroxyguanine (8-OHG), indicators of oxidative DNA damage, in tissue or body fluid are increased in diabetic patients. However, it is unclear whether plasma 8-OHG correlates with tissue 8-OHdG and whether insulin or antioxidant treatment reduces plasma 8-OHG in diabetic state. In this study, we measured the 8-OHG levels in plasma as well as the 8-OHdG levels in liver and kidney in streptozotocin-induced diabetic rats (DR) treated with insulin (DR+I), insulin and probucol (DR+I/P), or insulin and vitamin E (DR+I/E). There was a correlation between plasma 8-OHG levels and tissue 8-OHdG levels (plasma 8-OHG vs. liver 8-OHdG: r = 0.64, P < 0.001; plasma 8-OHG vs. kidney 8-OHdG: r = 0.38, P = 0.06). DR had levels of plasma 8-OHG that were three times higher than control rats (CR), whereas they had levels of tissue 8-OHdG that were approximately 1.5-2 times higher. Plasma 8-OHG levels in DR were almost normalized by insulin treatment, although insulin partially corrected hyperglycemia (plasma 8-OHG: CR 3.3 +/- 2.7 pmol/ml; DR 10.4 +/- 2.3 pmol/ml, P < 0.05 vs. CR; DR with insulin 3.6 +/- 1.0 pmol/ml, P < 0.05 vs. DR). However, tissue 8-OHdG levels in DR were significantly decreased by combined treatment with insulin and antioxidant (probucol or vitamin E), but not by insulin treatment alone. This data suggests that plasma 8-OHG could be a useful biomarker of oxidative DNA damage in diabetic subjects. The mechanism of differential response of plasma 8-OHG and tissue 8-OHdG to insulin and antioxidant treatment remains to be elucidated.     

氩氦刀冷冻消融与微波消融治疗原发性肝细胞癌近期疗效及对免疫功能的影响

目的 对比CT引导下氩氦刀冷冻消融与微波消融治疗原发性肝细胞癌（pHCC）近期疗效及对患者免疫功能的影响。方法 回顾性分析52例原发性pHCC患者,其中24例接受CT引导下氩氦刀冷冻消融（冷冻组）、28例接受CT引导下微波消融（微波组）,比较2组消融后1个月疗效,分析组间及组内消融前1日与消融后4周外周血CD4⁺、CD8⁺ T淋巴细胞占比及CD4⁺/CD8⁺的差异。结果 消融后1个月,冷冻组客观缓解率（87.50%）与微波组（85.71%）差异无统计学意义（P=0.833）。消融前1日,组间外周血CD4⁺、CD8⁺ T淋巴细胞占比及CD4⁺/CD8⁺差异均无统计学意义（P均>0.05）;消融后4周,组间外周血CD4⁺、CD8⁺ T淋巴细胞占比及CD4⁺/CD8⁺差异均有统计学意义（P均<0.05）,外周血CD4⁺ T淋巴细胞占比及CD4⁺/CD8⁺均较消融前1日升高（P均<0.001）、CD8⁺ T淋巴细胞占比较消融前1日降低（P均<0.001）。结论 氩氦刀冷冻消融与微波消融治疗pHCC近期疗效确切且相当,前者增强免疫功能的效果更佳。     

Prevalence and seasonal variation of hypovitaminosis D and its relationship to bone metabolism in community dwelling postmenopausal Hungarian women

Hypovitaminosis D can result in low bone mass. The prevalence of hypovitaminosis D has public health implications, especially where data are lacking. Since diet and sunlight are the two souces of vitamin D, the results obtained in one geographical region may not be universally applicable. The aim of this study is to characterize the prevalence and seasonal variation of hypovitaminosis D and its relationship to bone metabolism in community dwelling postmenopausal Hungarian women. We determined serum levels of 25-hydroxyvitamin D (25-OH-D), PTH, osteocalcin (OC), degradation products of C-terminal telopeptides of type-I collagen (CTx), dietary calcium intake and BMD at L2–L4 lumbar spine (LS) and femur neck (FN) in 319 randomly selected ambulatory postmenopausal women. The prevalence of hypovitaminosis D (serum 25-OH-D50 nmol/l) was 56.7%. On comparing patients with normal and low 25-OH-D, a significant difference was found in age (61.6±8.5 years versus 67.3±9.9 years; P<0.001), PTH (3.9±1.9 pmol/l versus 4.3±2.7 pmol/l; P<0.05), FN BMD (0.802±0.123 g/cm² versus 0.744±0.125 g/cm²; P<0.001) and dietary calcium intake (714.4±199.4 g/day versus 607.9±233 g/day; P<0.001). Osteoporotic patients had a significantly lower 25-OH-D (37.6±19.8 nmol/l versus 56.4±24 nmol/l; P<0.001) and dietary calcium intake (519.2±244.5 mg/day versus 718.2±164.3 mg/day; P<0.001). After controlling for all other variables, 25-OH-D was found to be significantly associated with age, the average hours of sunshine in the 3 months prior to 25-OH-D level determination and dietary calcium intake (r ²=0.190; P<0.001). For FN BMD, significant independent predictors were age, body mass index, 25-OH-D and dietary calcium intake (r ²=0.435; P<0.001). The prevalence of hypovitaminosis D during spring, summer, autumn and winter was 71%, 46.3%, 49.4% and 56.7%, respectively. There was significant seasonal variation in 25-OH-D, PTH, OC, calcium intake and FN BMD. There is a high prevalence of hypovitaminosis D in healthy postmenopausal Hungarian women, and FN BMD is associated with serum 25-OH-D and dietary calcium intake.     

Clinical significance of tumor-infiltrating lymphocytes in neoplastic progression and lymph node metastasis of human breast cancer

To investigate the clinical significance of tumor-infiltrating lymphocytes (TILs) within the tumor milieu, we quantitatively measured and compared the subpopulations of TILs in 24 patients with stage I–III breast carcinoma. Peripheral blood mononuclear cells (PBMCs), normal breast parenchyma-infiltrating lymphocytes (NILs), and TILs were isolated from tissue specimens and quantified by flow cytometry. The results showed that increased proportion of CD8⁺ T cells, with decreased proportion of CD4⁺ T cells, was significant in gated CD3⁺ TILs as compared to autologous NILs or PBMCs (P < 0.001). The tumor-infiltrating CD8⁺ T cells significantly increased with stage progression, reflected in a more strongly decreased CD4/CD8 percentage (P = 0.003). The CD4/CD8 percentage of TILs was strongly correlated with lymphovascular permeation and subsequent lymph node metastasis (P < 0.001). Increased percentages of tumor-infiltrating CD8⁺ T cells with decreased CD4/CD8 percentages are of prognostic importance for cancer progression in human breast cancer.     

Loss of Bone Mineral Density in Women Athletes During Aging

Total and regional bone mineral density (BMD) by dual-energy-X-ray absorptiometry (DXA) and bone turnover were tested in 50 highly trained women athletes and 21 sedentary control women (18–69 years; BMI < 25 kg/m²). VO_2max (ml · kg⁻¹· min⁻¹) and lean tissue mass (DXA) were significantly higher in the athletes versus controls (both P < 0.0001). Total body BMD did not decline significantly with age in the athletes whereas lumbar spine (L₂–L₄) BMD approached statistical significance (r =−0.26; P= 0.07). Significant losses of the femoral neck (r =− 0.42), Ward's triangle (r =−0.53), and greater trochanter BMD (r =−0.33; all P < 0.05) occurred with age in the athletes. In the athletes, total body BMD, L₂–L₄ BMD, and BMD of all sites of the femur were associated with lean tissue mass (r = 0.32 to r = 0.57, all P < 0.05) and VO_2max (r = 0.29 to r = 0.48, all P < 0.05). Femoral neck and greater trochanter BMD were higher in the athletes than in controls (both P < 0.05) and lumbar spine and Ward's triangle BMD approached statistical significance (both P= 0.07). Bone turnover was assessed by serum bone-specific alkaline phosphatase (B-ALP), urinary deoxypyridinoline cross-links (Dpd), and urinary aminoterminal cross-linked telopeptides (NTX). There were no relationships between B-ALP or Dpd with age whereas NTX increased with age (r = 0.46, P < 0.05) in the entire group. Levels of B-ALP and NTX were negatively associated with total body, L₂–L₄, femoral neck, Ward's triangle, and greater trochanter BMD (P < 0.05). B-ALP and Dpd were not significantly different between athletes and controls whereas NTX was lower in the athletes than in controls (P < 0.001). The high levels of physical activity observed in women athletes increase aerobic capacity and improve muscle mass but are not sufficient to prevent the loss of bone with aging. Received: 28 November 1997 / Accepted: 8 April 1998     

Oxidative DNA Damage Is Increased in Living Kidney Donors

BackgroundLong-term consequences of donor nephrectomy might be reduced kidney function, increased risk for cardiovascular disease, and impaired quality of life. The purpose of the current cross-sectional study was to evaluate the relationship between clinical, laboratory, and donation-specific outcomes of living kidney donors and systemic oxidative DNA damage.MethodsWe conducted a cross-sectional study and assessed retrospectively pre- and postdonation data from 60 donors who donated between 2010 and 2015. Plasma malondialdehyde levels and 8-hydroxy-2′-deoxyguanosine/deoxyguanosine ratio (8-OHdG/dG ratio) were determined as oxidative stress markers. Catalase, carbonic anhydrase, and paraoxonase (PON) activities were measured as antioxidants.ResultsApproximately 3 years after donation, the hypertensive donor ratio was 12%, and 11% of the donors had glomerular filtration rate <60 mL/min/1.73 m². Mean serum urea (P = .001) and serum creatinine levels (P = .001) were increased; creatinine clearance level (126.2 ± 35.5 vs 94.6 ± 26.8, P = .001) was decreased in the postdonation period. There was a significant positive correlation between predonation serum urea and 8-0HdG/dG ratio (r = 0.338, P = .016) and predonation serum creatinine and 8-0HdG/dG ratio (r = 0.442, P = .001), while there was a significant negative correlation between serum creatinine and PON activity (r = ?0.545, P < .001).ConclusionOur data have demonstrated that kidney donors exhibit increased oxidative DNA damage and decreased antioxidant activity. We propose that predonation serum creatinine is positively correlated with 8-0HdG/dG ratio and negatively correlated with antioxidant PON activity. This is the first study to demonstrate that plasma oxidative DNA damage increases in healthy kidney donors.     

The Na+-excreting efficacy of indapamide in combination with furosemide in massive edema

Background Massive systemic edema is often observed in patients with severe nephrotic syndrome, including diabetic nephropathy. Although furosemide, a loop diuretic, is often administered to these patients, some patients do not respond to this treatment, still showing massive edema.Methods The efficacy of indapamide which has a thiazide-like effect on distal convoluted tubules in combination with furosemide, was evaluated in eight patients with massive edema, in regard to both Na⁺ excretion and diuresis. Indapamide 2 mg was administered once a day, in the morning, to patients in whom it was considered that furosemide treatment of 40–120 mg a day for 1 week was ineffective.Results Urinary Na⁺ excretion was markedly increased, from 83.7 ± 82.2 mEq/day to 140.7 ± 33.8 mEq/day after 1 week of the combination therapy compared with furosemide alone (P < 0.01); urine volume was also increased, from 1070 ± 230 ml to 1359 ± 296 ml after 1 week of the combination therapy (P < 0.05). In this context, body weight was significantly decreased, from 57.2 ± 12.3 kg to 53.4 ± 12.8 kg, after the combination therapy (P = 0.01). Indapamide in combination with furosemide was well tolerated, and no significant changes in serum levels of creatinine and potassium were observed.Conclusions This combination therapy appears to be effective in patients with massive edema, as it increased diuresis, and achieved potent Na⁺ excretion.     

Immune profiling after minimally invasive lobectomy

Open in a separate window OBJECTIVESWhether acute phase and immune responses are minimally affected following minimally invasive lung surgery needs further investigation. We performed a pilot study to evaluate the immune profile of patients who underwent video-assisted thoracoscopic surgery or robot-assisted thoracic surgery lobectomies for the treatment of suspicious or known stage I non-small-cell lung cancer.METHODSBlood samples were taken preoperatively and 3 and 24 h postoperatively were analysed for C-reactive protein, glucose, cortisol, tumour necrosis factor alpha (TNF-α), interleukin 8 (IL-8) and interleukin 10 (IL-10) levels. TNF-α, IL-8 and IL-10 were also measured in lung tissues. T (CD4, CD8), B (CD19) and natural killer (CD56, CD16) cell counts and natural killer cell functions were analysed using a flow cytometry-based assay before and after surgery.RESULTSMinimally invasive surgery (robot-assisted thoracic surgery + video-assisted thoracoscopic surgery) significantly decreased IL-10 (P = 0.016) levels after surgery. No significant differences were detected in TNF-α (P = 0.48) and IL-8 (P = 0.15) levels before and after surgery. C-reactive protein (P < 0.001), cortisol (P < 0.001) and glucose levels (P < 0.001) increased significantly after surgery. Lymphocyte, total T cell, CD3⁺CD4⁺ and CD3⁺CD8⁺ CD16⁺CD56⁺ cell counts were significantly lower on postoperative day 1.CONCLUSIONThere seems to be a dynamic balance between pro- and anti-inflammatory cytokines and immune cells following minimally invasive lobectomy.     

The effects of calcium antagonists and prostaglandin El on isolated canine coronary arterial tension

The effects of calcium antagonists (nifedipine, nicardipine, diltiazem, and verapamil) and prostaglandin E₁ (PGE₁) on the tension of isolated canine coronary arterial strips were studies. In a solution containing 20 mEq/L of K⁺, 127 mEq/L of Na⁺, the tension was increased by 500–1,000 mg with 4 mEq/L of Ca²⁺. This increase in tension was suppressed by Ca-antagonists and PGE₁ dose-dependently. Nifedipine 10^–5 M, nicardipine 3 X 10^–7 M, diltiazem 3 X 10^–6 M, and verapamil 3 X 10^–6 M completely suppressed the increased tension. The maximal suppression of the tension produced by PGE₁ was about 40% at 10^–10 M. In 20 mEq/L K⁺ solution (0 mEq/L Ca²⁺, 37°C), the reduction of the Na⁺ concentrations from 127 mEq/L to 12 mEq/L increased the tension by 50 to 100 mg. This increase in tension was not suppressed by Ca-antagonists or PGEI. In conclusion, this study demonstrated that Ca-antagonists and PGE₁ suppressed an increase in the tension caused by Ca²⁺ but did not suppress an increase in the tension caused by Na⁺ reduction.     

Effect of menatetrenone on bone mineral density and incidence of vertebral fractures in postmenopausal women with osteoporosis: a comparison with the effect of etidronate

The purpose of the present study was to compare the effects of etidronate and menatetrenone on bone mineral density (BMD) and the incidence of vertebral fractures in postmenopausal women with osteoporosis. Seventy-two osteoporotic women, more than 5 years after menopause, 53–78 years of age, were randomly divided into three administration groups: E group; intermittent cyclical etidronate (200 mg/day, 14 days per 3 months; n = 25); M group; menatetrenone (45 mg/day, daily; n = 23); and C group (control); calcium lactate (2 g/day, daily; n = 24). Forearm BMD was measured by dual-energy X-ray absorptiometry at 0, 6, 12, 18, and 24 months after the treatment started. There were no significant differences in age, body mass index, years since menopause, and initial BMD among the three groups. One-way analysis of variance (ANOVA) with repeated measurements showed a significant decrease in BMD in the C group (P < 0.0001). Two-way ANOVA with repeated measurements showed a significant increase in BMD in the M group compared with that in the C group (P < 0.0001), and a significant increase in BMD in the E group compared with that in the C and M groups (P < 0.0001 and P < 0.01, respectively). The indices of new vertebral fractures/1000 patient-years in the E and M groups were significantly higher than that in the C group (χ² = 47.7; P < 0.0001 and χ² = 42.4; P < 0.0001, respectively), and did not differ significantly between the E and M groups. The present preliminary study provides evidence to suggest that, despite the lower increase in BMD produced by me-natetrenone, this agent, as well as etidronate, may have the potential to reduce osteoporotic vertebral fractures in postmenopausal women with osteoporosis. Received: January 9, 2001 / Accepted: June 8, 2001     

Diabetes and hepatic oxidative damage are associated with hepatitis C progression after liver transplantation

BACKGROUND: Posttransplant diabetes mellitus (PTDM) is common after liver transplantation and was recently identified as a risk factor for hepatitis C progression. Increased levels of oxidative stress have been identified in diabetes and hepatitis C. The aim of this study was to evaluate the relationship among PTDM, oxidative damage in liver biopsy specimens, and fibrosis progression posttransplant. METHODS: Subjects consisted of 27 hepatitis C-infected liver transplant recipients who had liver biopsy specimens available from 49 protocol liver biopsies. Paraffin embedded liver tissue sections were stained for 8-hydroxy-2' deoxyguanosine (8-OHdG), an indicator of hydroxyl radical mediated tissue damage. The percentage of cells staining for 8-OHdG in a histologic section was categorized as high (>66%) versus low score (< or =66%). Fibrosis index was calculated as fibrosis score (0-4)/ years posttransplant. Time to bridging fibrosis or cirrhosis (F3-4) was compared as a function of PTDM and 8-OHdG score. RESULTS: Considering all 49 biopsies, fibrosis index was higher in cases with PTDM (P<0.001) and high 8-OHdG score (P=0.004). High 8-OHdG score was associated with PTDM (P=0.012). In time to event analyses, time to F3-4 was more rapid in patients with PTDM (P=0.02) and in those with high 8-OHdG scores (P<0.001). CONCLUSIONS: This study confirmed a relationship between PTDM and hepatitis C fibrosis progression and found that oxidative damage in liver biopsy specimens was associated with PTDM and more rapid development of advanced fibrosis.     

Bacteremic shock: aspects of high-energy metabolism of rat liver following living Escherichia coli injection

Sublethal and lethal doses of living Escherichia coli (E. coli) (serotype: O-18) were injected intravenously into rats. The amounts of live E. coli injected were 0.5 ml of 0.8 – 1.0 × 10⁹ or 2.5 – 3.0 × 10⁹ organisms/ml saline per 100 g body weight for the sublethal (SL) or lethal (L) groups, respectively. The adenylate energy charge (ATP + 1/2ADP/ATP + ADP + AMP) and free NAD⁺/NADH ratios of the mitochondrial and cytoplasmic compartments in rat livers were measured at 3, 6, 12, and 24 hr following E. coli injection. At 6 hr in both groups, the hepatic energy charge levels were decreased from 0.856 in controls to 0.823 (SL) and 0.812 (L), associated with elevated mitochondrial NAD⁺/NADH ratios (P < 0.05). At 12 hr the sublethal group maintained the above state. In the lethal group, however, the hepatic energy charge was markedly decreased (P < 0.001), associated with remarkably decreased NAD⁺/NADH ratios in both mitochondria and cytoplasm (P < 0.01 and P < 0.001). In both groups the total ketone body (acetoacetate + β-hydroxybutyrate) contents were decreased with an elevation of mitochondrial NAD⁺/NADH. From these results, it is concluded that the difference between the sublethal and lethal groups is characterized by the progressive deterioration of the high-energy level in the lethal group, possibly due to restricted mitochondrial respiration. In addition, it is suggested that the elevated mitochondrial NAD⁺/NADH ratio may play a role in decreased ketone body contents in the liver following E. coli injection.     

When Is Curative Gastrectomy Justified for Gastric Cancer with Positive Peritoneal Lavage Cytology but Negative Macroscopic Peritoneal Implant?

For gastric cancer patients who have no peritoneal seeding at a macroscopic level but positive results in the peritoneal lavage cytology (PLC), the prognostic benefit expected by surgical resection is still controversial. During the period 1975–1994 as series of 417 consecutive patients without distant organ metastases underwent surgical resection for gastric cancer that had invaded the subserosal or deeper layers of the stomach wall. Immediately after laparotomy, the pouch of Douglas was washed with 100 ml of physiologic saline solution, and the fluid was collected for cytologic examination (four slide glasses) using Giemsa and Papanicolaou staining methods. According to the macroscopic (P) and cytologic (Cyt) results, the 417 patients were classified into three groups: P⁺ (n = 97); P⁻/Cyt⁺ (n = 25); and P⁻/Cyt⁻ (n = 295). Their 3-year survival rates after surgical resection were 4%, 24%, and 48%, respectively (p = 0.0001: P⁻/Cyt⁺ vs. P⁻/Cyt⁻; p = 0.0018: P⁻/Cyt⁺ vs. P⁺. Among the 25 P⁻/Cyt⁺ patients, postoperative survival was not associated with the T stage, N stage, cellular atypism, or cluster formation but with the number of cancer cells per slide during PLC. The 3-year survival rate was 35% for the subgroup with fewer than 10 cancer cells per slide (17 patients) and 0% for the other subgroup with 10 or more cancer cells per slide (8 patients) (p = 0.017). For P⁻/Cyt⁺ patients, who represent a subgroup of gastric cancer patients with an intermediate survival rate between the P⁻/Cyt⁻ and P⁺ patients, the number of cancer cells observed during PLC offers a potent prognostic indicator for the gastrectomy.     

Influence of ischaemia/reperfusion and LFA-1 inhibition on telomere lengths and CDKI genes in ex vivo haemoperfusion of primate kidneys

The telomere (T) length, p21^(WAF1/CIP1) and p27^(Kip1) cyclin-dependent kinase inhibitor (CDKI) genes are the markers of cell senescence and DNA damage. The aim of the study was to determine the influence of renal ischaemia/reperfusion (I/R) and anti-lymphocyte function-associated antigen-1 (LFA-1) monoclonal antibody (mAb) treatment on the value of the above-mentioned markers. Significantly higher levels of p21 and p27 were expressed by the glomeruli (P=0.001 and P=0.0001), tubules (P=0.0065 and P=0.0006), and interstitial cells (P=0.0017 and P=0.0022, respectively) of the xenoperfused kidneys. The mean T length of non-perfused renal specimens (5.56±0.60 kbp) was longer than that of the xenoperfused kidneys (5.46±0.36 kbp) [P= non-significant (NS)]. Addition of anti-LFA-1 mAb did not significantly influence the gene expression profile in the xenoperfused kidneys. The mean T length was longer in the kidneys with anti-LFA-1 mAb than in those without the medication (5.7±0.11 vs 5.13±0.31 kbp) (P=0.0661). Kidney I/R is associated with telomere shortening and an over-expression of p21 and p27 CDKIs, which indicates substantial DNA damage and/or accelerated tissue senescence. Although anti-LFA-1 mAb had some protective effect on the telomeres, it did not influence the gene expression profile in this study.A.B. Chkhotua and H. Schelzig share first authorship     

Urinary 8-hydroxydeoxyguanosine is elevated in patients with nephrolithiasis

8-hydroxydeoxyguanosine (8-OHdG) is an oxidatively modified guanosine, which has been widely used as an oxidative DNA damage marker in various diseases. The present study aimed to determine urinary 8-OHdG in nephrolithiasis patients and evaluate its clinical significance. Thirty-six nephrolithiasis patients and 30 healthy subjects were recruited. Urine volume, creatinine, malondialdehyde, β-N-acetylglucosaminidase (NAG) activity and proteins were measured in 24 h urine samples. Urinary 8-OHdG was determined by competitive enzyme-linked immunosorbent assay. Mineral composition of stones was analyzed using Fourier-transformed infrared spectroscopy. Nephrolithiasis patients excreted urinary 8-OHdG significantly higher than healthy controls. Urinary 8-OHdG levels compared among patients with calcium oxalate, struvite and uric acid stones were insignificantly different. The urinary NAG activity correlated positively with urinary 8-OHdG. Multiple linear regression showed that urinary NAG activity was an independent predictor of urinary 8-OHdG level. Receiver operating characteristic analysis revealed that the urinary 8-OHdG test was adequate for diagnosing nephrolithiasis. At 10 μg/g creatinine cutoff, the 8-OHdG test imparted high specificity (96.67%) and a positive predictive value (91.67%). In conclusion, this is the first report of elevated urinary 8-OHdG excretion in nephrolithiasis patients indicating increased oxidative DNA damage. Increased renal tubular damage was independently associated with elevated urinary 8-OHdG. Elevated urinary 8-OHdG levels adjunct with metabolic profile may be useful for identifying people at risk of stone development.     

Measurement of whole blood factor Xa-activated clotting time during hemodialysis with low-molecular-weight heparin

Purpose To determine whole blood factor Xa-activated clotting time (XaACT), a test for monitoring low-molecular-weight heparins (LMWHs). Methods Blood was obtained from six healthy volunteers. Dalteparin, a LMWH, was mixed with the blood to concentrations of 0,05 and 1.0IU·ml⁻¹. XaACT, activated clotting time (ACT), and activated partial thromboplastin time (APTT) were measured at each dalteparin concentration. XaACT of blood from the outflow and inflow sides of the blood circuit in seven hemodialysis patients was measured before and after bolus administration of 1000 IU of dalteparin, followed by continuous infusion at a rate of 500IU·h⁻¹. Results XaACT, ACT, and APTT in dalteparin-containing blood from volunteers were correlated with dalteparin concentration (y=312.8x+86.4;r ²=0.88;P<0.001,y=41.8x +113.5;r ²=0.83;P<0.001, andy=59.5x+38.8;r ²=0.80;P<0.001, respectively). The regression slope of XaACT was steeper than those of ACT and APTT (P<0.001). In hemodialysis patients, dalteparin increased XaACT on the outflow and inflow sides of the circuit (P<0.001,P<0.05, respectively). Conclusion The measurement of XaACT can be employed to monitor LMWHs in clinical settings.