Urinary Alkaline Phosphatase Research Articles

Cadmium exposure in tobacco workers: possible renal effects

Cadmium is a nephrotoxic metal widely used in industry and the main source of Cd in general population is smoking. Considering that the source of Cd in cigarettes is the tobacco leaf, the exposure to Cd was evaluated in workers employed at a tobacco leaf processing factory. Blood and urinary Cd levels were measured by flameless atomic absorption spectrometry in 87 workers and 35 controls. Urinary enzymes, total protein, albumin and uric acid were also determined to investigate the possible nephrotoxic effects of Cd. Blood Cd levels were significantly higher in workers (1.63 ± 1.95 μg/L) than in controls (0.91 ± 1.15 μg/L) (p = 0.044). The increase observed in urinary Cd levels of workers was non significant (0.56 ± 0.5 μg/g creatinine in workers and 0.46 ± 0.5 μg/g creatinine in controls). Both in workers and in controls, subjects smoking >10 cigarettes/day showed significantly increased blood Cd levels compared to non-smokers (p = 0.000 and p = 0.011, respectively). In workers, urinary alkaline phosphatase (ALP), gamma glutamyl transferase (GGT), total protein, and uric acid were observed to be significantly increased (p = 0.013, p = 0.000, p = 0.000, p = 0.025, respectively), ALP, GGT and total protein being positively correlated with Cd in urine. In conclusion, the workers in the tobacco leaf processing factory were found to be exposed to Cd compared to the general population. The increase in the urinary enzymes and proteins suggests that an exposure to Cd affects kidney functions even below the toxic limits generally accepted.

Liver and kidney function and histology in rats exposed to cadmium and ethanol.

The present study was performed to assess the function and histology of the liver and kidney in rats exposed to 50 mg Cd/l (as cadmium chloride) and/or 10% (w/v) ethanol (EtOH) for 12 weeks. The activities of alanine aminotransferase (ALAT) and asparate aminotransferase (AspAT) in serum were measured as indicators of the liver function. As parameters of the kidney function, creatinine, total protein and urea concentrations in serum and urine, as well as urinary alkaline phosphatase (ALP) activity were determined, and creatinine clearance was calculated. Both organs were subjected to histopathological analysis. Daily Cd intake ranged from 3.17 to 4.28 mg/kg body weight and from 2.41 to 3.17 mg/kg body weight in the Cd and Cd + EtOH groups, respectively. The daily intake of 10% EtOH ranged from 47.5 to 86.9 g/kg body weight in the EtOH and from 47.3 to 63.4 g/kg body weight in the Cd + EtOH-exposed rats. Cd and EtOH, independently of separate or combined application, changed liver and kidney function and histology. Rats treated with Cd alone and those co-exposed to both substances showed qualitatively similar, but different magnitudes of changes, in liver and kidney histology. Blurred trabecular structure, vacuolar degeneration and increased density of nuclear chromatin with very compact nuclear structure were found in hepatocytes of zones 2 and 3. Moreover, mononuclear cell infiltrations and necrosis of single cells were evident in zone 1. In the kidney tubules, degeneration and hypertrophy of epithelial cells and dilation in the glomeruli were also observed. Some functional (increased serum AspAT and urinary ALP, decreased urinary urea) and structural changes in the liver and kidney were more evident in the case of combined exposure, while others were more evident after single exposure. However, a decrease in creatinine clearance, noted only in the animals treated with Cd and EtOH, shows that functional changes indicating renal insufficiency are more serious in the co-exposed group. Due to lower Cd and EtOH intake (resulting from a stronger aversion to drinking water containing both substances) in the co-exposed rats, as compared to the Cd- and EtOH-treated groups, it is difficult to draw a definite conclusion from this study. The findings, however, seem to indicate that EtOH increases Cd nephrotoxicity in rats, and thus may suggest a higher risk of kidney damage in alcoholics exposed to Cd. Unfortunately, this study does not provide clear evidence if, and to what extent, EtOH influences Cd hepatotoxicity.

Role of plasma and urinary calcium and phosphorus measurements in early detection of phosphorus deficiency in very low birthweight infants.

To analyse the role of serum and urinary calcium and phosphorus levels in early detection of mineral deficiency in very low birthweight (VLBW) infants born appropriate (AGA) and small for gestational age (SGA). 64 VLBW infants were included in a cohort study and divided into two groups: AGA (n = 30) and SGA infants (n = 34). Then, they were divided according to the presence of radiological signs of metabolic bone disease (MBD): with MBD (n = 21) and without MBD (n = 34). Blood samples and 6 h urine collections were obtained for calcium, phosphorus, alkaline phosphatase activity and creatinine determinations between 3 and 5 wk of life. There were no biochemical differences between AGA and SGA. Higher values of urinary calcium (MBD = 31.9 +/- 20.2, without MBD = 19.8 +/- 15.4; p = 0.017), calciuria (MBD = 2.3 +/- 0.3, without MBD = 1.4 +/- 0.8; p = 0.037) and alkaline phosphatase activity (MBD = 369 +/- 114, without MBD = 310 +/- 93; p = 0.04) were found in infants who developed MBD. Both groups showed high tubular phosphorus reabsorption indicating mineral deficiency. Serum calcium and phosphorus levels are not good markers in early detection of mineral deficiency. However, the monitoring of calcium urinary levels may be helpful in early detection of mineral deficiency.

Glomerular and tubular renal functions after long-term medication of sulphasalazine, olsalazine, and mesalazine in patients with ulcerative colitis.

To date there are only few reports evaluating the potential nephrotoxic reactions of the new 5-aminosalicylic acid (5-ASA) preparations in patients with ulcerative colitis (UC). The aim of this study was to screen the tubular and glomerular functions in patients with UC in maintenance treatment with either 5-ASA azo-compounds (sulphasalazine and olsalazine) or mesalazine. Patients with UC in clinical remission treated with either sulphasalazine, olsalazine, or mesalazine for more than 1 year were included in an open, single-blind retrospective Norwegian multicenter study. Serum and urine creatinine, serum and urine beta2-microglobulin, urine N-acetyl-beta-glucoseamidase (NAG), urine alkaline phosphatase, urine microalbumin, urine alanine amino peptidase, and urine beta2-microglobulin were measured. Fifty-two females and 75 males (n = 127), ages 20-69, were evaluated. Thirty-six patients were treated with sulphasalazine (mean treatment time 10.1+/-6.6 years [mean +/- SD]), 32 patients were treated with olsalazine (2.3+/-1.4 years), and 59 patients with mesalazine (3.2+/-2.0 years). At inclusion, there were no significant differences in the serum or urine values between the groups. In 17 patients (1 patient [3%] in the sulphasalazine group, 4 patients [13%] in the olsalazine group, and 12 patients [20%] in the mesalazine group), at least one abnormal serum and/or urine value was detected. After 10 years of treatment, only one abnormal value was found among the 19 patients in the sulphasalazine group. The abnormal values observed in the other groups indicated minor glomerular or tubular renal damage. In conclusion, long term sulphasalazine treatment appears to be safe and free of nephrotoxic side effects, whereas minor glomerular and tubular impairment are observed in a few patients treated with olsalazine and mesalazine.

Effect of thiamine on the cadmium-chelating capacity of thiol compounds.

The influence of thiamine on the efficacy of meso-2,3-dimercaptosuccinic acid (DMSA), diethyldimercapto succinate (DEDMS), alpha mercapto-beta-(2-furyl) acrylic acid (MFA) and alpha-mercapto-beta-(2-thienyl) acrylic acid (MTA) to mobilize cadmium and reverse cadmium-induced biochemical alterations was investigated in cadmium-exposed rats. The thiamine coadministration enhanced the efficacy of MFA and MTA in reducing hepatic and renal burden of cadmium and that of DMSA and DEDMS in mobilizing hepatic cadmium. It also improved the efficacy of DMSA, DEDMS and MFA in reversing the cadmium-induced increase in urinary alkaline phosphatase and aspartate and alanine amino transaminases. The combined treatment with thiamine and DMSA or MFA restricted the urinary loss of zinc and that with thiamine and DEDMS reduced the loss of fecal copper, a general effect of chelation. In conclusion, the administration of thiamine during chelation therapy in cadmium poisoning may be beneficial and more effective than thiol chelating agents alone, which needs to be confirmed in humans.

Effects of general anesthesia and surgery on renal function in healthy dogs.

To evaluate renal function in healthy dogs undergoing general anesthesia and ovariohysterectomy without concurrent IV administration of fluids. 35 healthy client-owned dogs. Dogs were medicated with promazine hydrochloride (0.05 mg/kg of body weight, SC) approximately 45 minutes before induction of anesthesia with thiopental sodium (10 to 15 mg/kg, IV). Anesthesia was maintained with 2% halothane in oxygen. Ovariohysterectomies were performed by senior veterinary students under the direct supervision of a veterinary surgeon. Renal function was assessed (serum urea and creatinine concentrations, fractional clearance of sodium, urine alkaline phosphatase [ALP] and gamma-glutamyltransferase [GGT] activities, urine specific gravity, and enumeration of renal tubular epithelial cells in urine sediment) prior to and 24 and 48 hours after surgery. Duration of general anesthesia ranged from 80 to 310 minutes. Urine specific gravity and ALP activity and serum urea and creatinine concentrations did not change over time. Fractional clearance of sodium decreased 24 and 48 hours after surgery, whereas urine GGT activity and the ratio of urine GGT activity to urine creatinine concentration increased 24 hours after surgery, compared with presurgery values. Renal tubular epithelial cells increased in number in urine sediment from 11 of 35 (31.4%) dogs and 5 of 35 (14.3%) dogs 24 and 48 hours after surgery, respectively. However, this increase was not clinically relevant. Intravenous administration of fluids to healthy dogs undergoing general anesthesia and elective surgery may not be necessary for maintenance of renal homeostasis.

Determination of Urinary and Serum β-Glucuronidase and Alkaline Phosphatase in Various Renal Disease and Kidney Rejection Transplanted Patients

ABSTRACT β-Glucuronidase (β-Glu) and alkaline phosphatase (ALP) were evaluated in serum and urine in 50 subjects classified into five equal groups. Group I was control healthy subjects, while groups II, III, IV, and V were patients with nephritic syndrome, pyelonephritis, kidney rejection, and end-stage renal disease, respectively. Urinary β-Glu was significantly elevated in all four groups; while serum enzyme showed no change. On the other hand, serum ALP showed a significant elevation in all abnormal groups. Accordingly, urinary β-Glu and serum ALP could be used as diagnostic markers for various renal diseases.

Spironolactone, But Not Flutamide, Administration Prevents Bone Loss in Hyperandrogenic Women Treated With Gonadotropin-Releasing Hormone Agonist

GnRH agonists (GnRHa) have recently been proposed for the treatment of hirsutism in women with the polycystic ovary syndrome (PCOS). As most of these subjects have increased androgen secretion from both ovaries and adrenal glands, the association of GnRHa with antiandrogen drugs might enhance the clinical response to treatment. On the other hand, this association might also potentiate the adverse effects of GnRHa on bone metabolism, generating a potentially harmful situation at the skeletal level. In this study we investigated in 41 PCOS patients the skeletal effects of a 6-month course of GnRHa (tryptorelin, 3.75 mg, im, monthly), either alone (n 5 12) or associated with the antiandrogen drugs spironolactone (100 mg, orally, once daily; n 5 14) or flutamide (250 mg, once daily; n 5 15). In all subjects bone mineral density was measured before and after treatment by dual energy x-ray absorptiometry at the lumbar spine (L2‐L4) and at the femoral neck and Ward’s triangle. In addition, at baseline and after 6 months of therapy, bone metabolism markers (serum and urinary calcium, serum phosphorus and alkaline phosphatase, plasma osteocalcin, and urinary hydroxyproline) and endocrine parameters (serum gonadotropins, estradiol, and free testosterone) were assayed. Women given either GnRHa alone or associated with spironolactone or flutamide were similar for age and body mass index. At baseline, the 3 groups of PCOS women were also similar for endocrine and bone parameters. After 6 months, all treatments determined similar striking suppressions of serum gonadotropins and sex steroids. Concurrently, bone mineral density was significantly reduced at all examined sites in subjects receiving either GnRHa alone or GnRHa plus flutamide. Conversely, women given GnRHa plus spironolactone did not show any change in skeletal mass from baseline values (P , 0.05‐ 0.01 among groups). Biochemical parameters of bone metabolism were consistent with densitometric assessments. In conclusion, after a 6-month course of therapy, bone mineral density is reduced in PCOS women given either GnRHa alone or GnRHa plus flutamide, but not in those receiving GnRHa plus spironolactone. The mechanisms of the bone-sparing effect of spironolactone remain to be determined. Nevertheless, this drug could represent a useful tool to prevent skeletal loss in women given GnRHa as well as in other hypoestrogenic conditions, in particular when estrogens are not recommended. (J Clin Endocrinol Metab 84: 1250 ‐1254, 1999)

Benzene exposure in car mechanics and road tanker drivers.

The purpose of this study was to identify professional factors related to benzene exposure and to deduce suitable safety measures. Atmospheric benzene, urinary muconic acid (tt-MA) and leukocyte alkaline phosphatase activity (LAPA) were evaluated among 66 car mechanics, 34 road tanker drivers, and 28 nonexposed workers. Professional and medical questionnaires were filled in at the same time. Atmospheric benzene was significantly higher among road tanker drivers than among car mechanics. The arithmetic mean +/- SD, median, and geometric mean values were, respectively, 0.48 +/- 1.49, 0.14, and 0.06 mg/m3 among car mechanics and 1.88 +/- 4.18, 0.68, and 0.65 mg/m3 among road tanker drivers. In the latter case the increase was caused by transport of unleaded petrol and correlated with the volume of the tank. Among car mechanics, tobacco smoking, windy conditions, dismantling of petrol filters, and handling of petrol increased atmospheric benzene levels. Urinary muconic acid was increased significantly among car mechanics (148 +/- 137, 127, and 111 micrograms/g) and among road tanker drivers (309 +/- 420, 137, and 151 micrograms/g) as compared with the controls (49 +/- 46, 33, and 33 micrograms/g). Among road tanker drivers, alcohol intake and transportation of unleaded petrol increased the excretion of muconic acid, which was also directly related to the volume of the tank. Among car mechanics, professional factors (dismantling of petrol filters, handling of and washing of hands with petrol) and nonprofessional factors (tobacco smoking and damaged skin on the hands and forearms) increased muconic acid excretion. In the control group, tobacco smoking increased its excretion. LAPA was not significantly modified among exposed workers. There was a weak but significant linear correlation between LAPA and muconic acid. These results suggest that to reduce exposure to benzene in unleaded petrol, individual and collective safety measures should be imposed in both occupations.

Cytotoxicity and cell-proliferation induced by the nephrocarcinogen hydroquinone and its nephrotoxic metabolite 2,3,5-(tris-glutathion-S-yl)hydroquinone.

Hydroquinone, an intermediate used in the chemical industry and a metabolite of benzene, is a nephrocarcinogen in the 2-year National Toxicology Program bioassay in male Fischer 344 rats. Current evidence suggests that certain chemicals may induce carcinogenesis by a mechanism involving cytotoxicity, followed by sustained regenerative hyperplasia and ultimately tumor formation. Glutathione (GSH) conjugates of a variety of hydroquinones are potent nephrotoxicants, and we now report on the effect of hydroquinone and 2,3,5-(tris-glutathion-S-yl)hydroquinone, on site-selective cytotoxicity and cell proliferation in rat kidney. Male Fischer 344 rats (160-200 g) were treated with hydroquinone (1.8 mmol/kg or 4.5 mmol/kg, p.o.) or 2,3,5-(tris-glutathion-S-yl)hydroquinone (7.5 micromol/kg; 1.2-1.5 micromol/rat, i.v.), and blood urea nitrogen (BUN), urinary gamma-glutamyl transpeptidase (gamma-GT), alkaline phosphatase (ALP), glutathione-S-transferase (GST) and glucose were measured as indices of nephrotoxicity. Hydroquinone (1.8 mmol/kg, p.o.) is nephrotoxic in some rats, but not others, but cell proliferation (BrDU incorporation) in proximal tubular cells of the S3M region correlates with the degree of toxicity in individual rats. At 4.5 mmol/kg, hydroquinone causes significant increases in the urinary excretion of gamma-GT, ALP and GST. Pretreatment of rats with acivicin prevents hydroquinone-mediated nephrotoxicity, indicating that toxicity is dependent on the formation of metabolites that require processing by gamma-GT. Consistent with this view, 2,3,5-(tris-glutathion-S-yl)hydroquinone, a metabolite of hydroquinone, causes increases in BUN, urinary gamma-GT and ALP, all of which are maximal 12 h after administration of 2,3,5-(tris-glutathion-S-yl)hydroquinone. In contrast, the maximal excretion of GST and glucose occurs after 24 h. By 72 h, BUN and glucose concentrations return to control levels, while gamma-GT, ALP and GST remain slightly elevated. Examination of kidney slices by light microscopy revealed the presence of tubular necrosis in the S3M segment of the proximal tubule, extending into the medullary rays. Cell proliferation rates in this region were 2.4, 6.9, 15.3 and 14.3% after 12, 24, 48 and 72 h, respectively, compared to 0.8-2.4% in vehicle controls. Together with the metabolic data, the results indicate a role for hydroquinone-thioether metabolites in hydroquinone toxicity and carcinogenicity.

Clinical significance of urinary human tissue non-specific alkaline phosphatase (hTNAP) in pre-eclampsia and eclampsia.

The aim of this work was to determine the levels of urinary human tissue non-specific alkaline phosphatase (hTNAP) in pre-eclampsia and eclampsia in order to assess renal tubular damage. Urine samples were collected from 26 mild pre-eclamptic, 26 were pre-eclamptic, 20 eclamptic patients and 20 healthy pregnant women (controls) in their late third trimester. Urinary hTNAP/creatinine (hTNAP/cr) in severe pre-eclampsia and eclampsia were significantly higher than in controls. Urinary hTNAP/cr was increased in 23%, 77% and 90% of cases of mild pre-eclampsia, severe pre-eclampsia and eclampsia, respectively, indicating that the increase correlates with the severity of the disease. Marked elevation or urinary hTNAP/cr was also associated with bad fetal outcome. These results provide additional evidence for renal tubular damage in pre-eclampsia and eclampsia.

Renal tubular dysfunction in patients with inflammatory bowel disease treated with aminosalicylate.

An increasing number of case reports indicate potential nephrotoxicity of 5-aminosalicylic acid (5-ASA), which shares similarities with the chemical structures of both phenacetin and acetylsalicylic acid. In a point prevalence study the occurrence of sensitive indices indicative of early kidney malfunction was assessed in outpatients with inflammatory bowel disease. Routine indices of kidney function (creatinine clearance, urinary protein content, pH, electrolytes, and microscopy) were investigated in 223 patients with inflammatory bowel disease as well as sensitive markers of glomerular or tubular dysfunction (microproteinuria by SDS polyacrylamide gel electrophoresis (SDS-PAGE), urinary concentrations of N-acetyl-beta-D-glucosaminidase, alpha 1-microglobulin, gamma-glutamyltransferase (GGT), alkaline phosphatase (AP), and albumin). Histories of exposure to 5-ASA were assessed by questionnaire. Patients receiving high amounts of 5-ASA, both actual as well as on a lifetime basis, showed an increased prevalence of tubular proteinuria by SDS-PAGE. Raised values for urinary AP and GGT indicate proximal tubular epithelial cells as the source. All other kidney function tests were normal. Analysis of covariates indicated strong associations between disease activity and size of 5-ASA doses as well as alterations in kidney tubular function. The possibility exists that high doses of 5-ASA may be associated with proximal tubular proteinuria. This point prevalence study cannot dissect the possible impact of chronic inflammation from high dose 5-ASA treatment and further prospective studies are warranted.

Comparative biochemical effects of low doses of mercury II chloride in the F344 rat and the multimammate mouse ( Mastomys natalensis)

The biochemical effects and comparative nephroxicity of mercury II chloride (HgCl 2) dosed at 0.75 mg/kg i.p. was investigated in the Fisher 344 rat (F344) and Mastomys natalensis using high resolution 1H nuclear magnetic resonance (NMR) spectroscopy of urine, histopathology and clinical chemical techniques. The effects of HgCl 2 treatment were followed for up to 4 days post-dosing (p.d.). In F344 rats there was extensive proximal tubular damage and renal cortical necrosis together with elevated levels of urinary γ-glutamyl transpeptidase (γGT), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH). The 1H NMR spectra of urine obtained from Hg-treated F344 rats also showed increased levels of glucose, alanine, lactate, valine and hippurate (0–48h p.d.) with decreased levels of citrate, succinate and 2-Oxoglutarate (24–48h p.d.). Mastomys were found to be highly resistant to HgCl 2 toxicity at 0.75 mg/kg and the histological appearance of the renal cortex of treated animals was virtually identical to controls. There were no elevations in urinary ALP, γGT and LDH activities in HgCl 2-treated Mastomys and there were no biochemical abnormalities in low MW components of Mastomys urine following HgCl 2-treatment, as shown by 1H NMR spectroscopy. Urinary γGT activity was found to be much higher in F344 rats than Mastomys. Since γGT activity is involved in the tubular reabsorption of Hg 2+, the lower levels of γGT in Mastomys might partially account for the lower toxicity of Hg 2+ in this species.

Some urinary enzymes in bilharziasis.

Urinary alkaline phosphatase (ALP), acid phosphatase (ACP), aryl sulphatase (Ar. sulph.), beta-glucuronidase (beta-gluc.) and galactosidase were assayed in a group of Bilharzia haematobium patients and another group of healthy subjects (control group). The results for most of the determined enzymes revealed high activities as compared to the controls. The activity of acid phosphatase in male urine samples increased also, though not significantly. These elevated enzyme activities could be used to establish the diagnosis of schistosomiasis in patients whose urine contains no ova or when it is difficult to detect them. The results are discussed in the light of localization of each enzyme in the urinary tract as well as in other organs like the liver.

Incidence and prediction of stone recurrence after lithotripsy in idiopathic calcium stone patients: a multivariate approach.

In order to evaluate the recurrence of calcium kidney stones, 520 patients (275 males and 245 females), aged 14-79 years, previously treated with lithotripsy were followed up for 23 months on average (median = 24 months; range = 12-48 months), and 101 relapses (10%/year) were recorded. Among the possible predictors of recurrence, measured at the beginning of the follow-up and analyzed in univariate and multivariate statistical ways, age was inversely related with occurrence of event (multivariate t value = -2.12) whereas urinary calcium (UC; t = 2.78), alkaline phosphatase (AP; t = 3.55) and history of previous relapses (t = 2.07) were directly related to the recurrence. The levels of UC were not correlated to those of AP (linear correlation coefficient r = 0.0032), but the combination of their high levels increased the risk of recurrences. The contribution of the other considered factors to stone formation were not significant (sex, family history of stone disease, gallstone, renal failure, serum calcium, phosphate, uric acid, sodium and proteins, urinary phosphate, sodium, magnesium and uric acid.

ESTIMATING BONE CHANGE IN PATIENTS WITH SEVERE DISABILITY

To evaluate bone change in severely disabled patients, bone density was assessed on X-ray pictures by the microdensitometry (MD) method, and urinary hydroxyproline (U-HYP), urinary glycosaminoglycans (U-GAG) serum calcium (S-Ca) and serum alkaline phosphatase (S-AP) were assessed in 43 patients (mean age 16 years) at the National Sanatrium Ehime Hospital. On division of the severity of bone change into grades 0 to 3 using the MD method. U-HYP was found to be significantly higher in grades 2 and 3 than in controls. Tendencies were similar for U-GAG and S-AP. U-HYP in immobile patients was significantly higher than in mobile patients. Immobilisation was one of the most important contributors to the development of bone change and results obtained using the MD method, U-HYP and U-GAG were valuable-indices of the bone change in severely disabled patients.

Significance of elevated urinary human intestinal alkaline phosphatase in Japanese people exposed to environmental cadmium

Urinary human intestinal alkaline phosphatase (IAP), β 2-microglobulin ( β 2-MG) and N-acetyl- β- d-glucosaminidase (NAG) were analyzed in 40 Japanese environmental-cadmium (Cd)-exposed and 40 non-exposed subjects to evaluate early biological markers for Cd-induced renal damage. All urinary indicators were significantly higher in the Cdexposed subjects than non-exposed subjects. A fourth-order function was fitted for the relationship between β 2-MG and IAP or NAG. The β 2-MG concentration corresponding to the inflexion point for IAP was smaller than that for NAG. This result may support the contention that the cells containing IAP are damaged earlier than those containing NAG, and that IAP is a useful marker for detecting renal tubular dysfunction in people moderately exposed to Cd. However, in the stage of severe renal damage, the combination of IAP and β 2-MG is considered to be more useful.

Increased urinary excretion of pyridinium cross-links as markers of bone resorption in bone metastases of lung cancer

Pyridinoline (Pyd) and deoxypyridinoline (D-pyd), two collagen-based cross-links found in bone, were measured by high-performance liquid chromatography (HPLC) in urine samples from 17 male control subjects and 17 male lung cancer patients either with or without bone metastases. Ten patients with bone metastases had significantly higher (P < 0.01) urine concentrations of Pyd and D-pyd than did control subjects. No cross-link increase was seen in seven lung cancer patients without clinically detectable bone metastases. We also measured contemporaneously urinary hydroxyproline (OH-Pro) and serum alkaline phosphatase (ALP). We found higher concentrations of OH-Pro and ALP in patients with metastatic lung cancer compared to those with nonmetastatic lung cancer or control subjects. There was a significant correlation among pyridinium cross-links, OH-Pro, and ALP. In addition, the excretion of pyridinium cross-links in lung cancer with metastases correlated directly with bone scan Soloway scores (rs = 0.74,P = 0.02;rs = 0.67,P = 0.03, respectively), whereas levels of OH-Pro and the ALP level did not (rs = 0.52,P = 0.07 andrs = 0.50,P = 0.13, respectively). These findings suggest that the measurement of Pyd and D-pyd in urine may be used to quantitate bone metastases in lung cancer.

Enhanced hexachloro-1:3-butadiene nephrotoxicity in rats with a preexisting adriamycin-induced nephrotic syndrome.

Renal damage was assessed by histopathology and urinalysis in male Wistar rats treated with either hexachloro-1:3-butadiene (HCBD; a single 170-mg/kg ip dose that caused proximal tubule necrosis), adriamycin (ADR; a single 5-mg/kg ip dose that caused minimal glomerular changes up to 35 days), or HCBD given 2 wk after ADR and compared with age-matched control rats for 21 days. Urinalysis values in ADR-treated rats showed minimal renal changes. HCBD significantly elevated urine volume (10-fold), protein (5-fold), glucose (175-fold), and brush border enzymes (10-600-fold), indicating severe proximal tubular damage, but most parameters returned to pretreatment levels 6 days after treatment. In ADR-pretreated rats subsequently given HCBD, both the urinary alkaline phosphatase and the ratio of kidney: body weight were significantly higher for longer periods. Histopathology demonstrated that the HCBD-induced proximal tubular lesion was confined to the outer stripe of the outer medulla. Advanced regeneration and repair was evident 21 days after HCBD treatment. In the ADR-pretreated rats the HCBD-induced lesion was more severe and affected the entire cortex and was characterized by marked tubular epithelial calcification, with little evidence of repair and tubular restitution 21 days after treatment. Enzyme histochemistry showed gamma-glutamyltranspeptidase localized to the proximal tubules. After HCBD treatment the enzyme staining was lost and subsequently returned in parallel with histological recovery up to 21 days. The distribution and intensity of gamma-glutamyltranspeptidase was unchanged in ADR-treated rats. The distribution and intensity of gamma-glutamyltranspeptidase in kidneys of ADR-pretreated rats given HCBD had not returned to normal by day 21. The results of this study indicate that pretreatment with ADR increases HCBD-induced nephrotoxic damage and decreases renal cortical repair capacity.

Early Morphological and Biochemical Changes During 2-Br-(diglutathion-S-yl)hydroquinone-Induced Nephrotoxicity

Early subcellular targets of 2-Br-(diglutathion-S-yl)hydroquinone (2-Br-(diGSyl)HQ)-mediated nephrotoxicity were investigated by morphological and biochemical criteria. After treatment of male Fischer 344 rats with 2-Br-(diGSyl)HQ (30 μmol/ kg), proximal tubular morphology was examined by electron microscopy. Changes in the plasma membrane, nuclei, and endoplasmic reticulum were observed within 30 min of 2-Br-(di-GSyl)HQ administration. These changes consisted of loss of the brush border membrane, margination of heterochromatin, and reorganization of the endoplasmic reticulum into discrete aggregates. The desquamation of the brush border membrane into the tubular lumen corresponded with the rapid excretion of γ-glutamyl transpeptidase and alkaline phosphatase in urine. As the injury developed, cell swelling with loss of cytosolic density and loss of chromatin staining was observed, and between 2 and 4 hr the nuclei underwent extensive karyorrhexis and karyolysis. Agarose gel electrophoresis of DNA isolated from the corticomedullary junction at 4 hr exhibited extensive fragmentation, which was random in nature. Mitochondria assumed a condensed configuration 2 hr after 2-Br-(diGSyl)HQ administration, but this was not followed by high-amplitude swelling prior to cell death and necrosis. Biochemical assessment of mitochondria, isolated from 2-Br-(diGSyl)HQ-treated rats at 2 hr, exhibited a significant (20%) decrease in respiratory control ratios (RCR), a consequence of an increase in State 4 respiration. At later time points (8 hr) State 4 respiration returned to control values, but the respiratory control ratio (RCR) remained significantly depressed due to decreases in State 3 respiration. At this time blood urea nitrogen concentrations were significantly elevated (41 ± 3, mean ± SD, n = 10). The data suggest that the plasma membrane and the nucleus are early targets of 2-Br-(diGSyl)HQ-induced cytotoxicity, and that alterations in mitochondrial structure and respiratory function occur following the initial injury.