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Biomedical subjects

E Bergamaschi

Publications and source records attributed to E Bergamaschi.

At least 19 recordsLinked to original sources

Effects of urinary macromolecules on the nucleation of calcium oxalate in idiopathic stone formers and healthy controls.

Urinary macromolecules have attracted great interest because of their possible role as both promoters and inhibitors of calcium oxalate (CaOx) crystallization and it remains unclear whether there is any difference, in their nucleating activity, between stone formers and controls. We selected 9 male idiopathic CaOx stone formers whose 24-h urines presented no evidence of common urinary stone risk factors such as hypercalciuria, hyperoxaluria, hyperuricosuria, hypocitraturia, hypomagnesiuria or low glycosaminoglycans excretion and 12 male controls (matched for age and body weight) whose 24-h urines did not differ from those of stone formers. The study of urinary CaOx nucleation was made in freshly voided overnight urines whose biochemical composition was almost identical in the two groups. In filtered (0.22 micron) and ultrafiltered (10 kDa) urine we performed an oxalate tolerance test to determine the permissible increment of oxalate, the oxalate level for nucleation and the permissible increment of CaOx relative supersaturation (CaOx RS). In filtered urine from stone formers the permissible increment of oxalate was lower than controls (30 +/- 10.2 vs. 46.7 +/- 9.7 mg/l, P = 0.001), the oxalate level for nucleation was lower (64.4 +/- 14.2 vs. 79.5 +/- 15.6 mg/l, P = 0.035) and the permissible increment of CaOx RS was also lower (9.71 +/- 2.59 vs. 13.39 +/- 3.62, P = 0.018). In ultrafiltered urine these differences disappeared because the removal of macromolecules in stone formers significantly enhanced the oxalate-tolerance values. The difference between the change of the oxalate permissible increment of filtered and ultrafiltered urine allowed a distinction to be made between stone formers and controls that was not feasible in other ways (7.6 +/- 5.3 vs. 3.3 +/- 5.9 mg/l, P < 0.0001). The study suggests that, in idiopathic CaOx stone formers free from common urinary risk factors of CaOx crystallization, there is an increased tendency for CaOx nucleation in urine, which is mediated by macromolecular components.

Adult

Immunological changes among workers occupationally exposed to styrene.

The functional status of the immune system was investigated in a group of 71 workers exposed to styrene and in 65 control subjects, recruited according to the same selection criteria and comparable as to sex, age, and confounding variables. Air and biological monitoring were used to characterize styrene exposure (median of the main urinary metabolites in the "next-morning" spot samples: 106 mg/g creatinine). Phenotypic analysis of peripheral blood lymphocytes (PBL) by automated flow cytometry revealed a reduced proportion of T lymphocyte subsets (CD3+, CD4+ and CD4+45+), with no changes in CD8+, and a higher proportion of B lymphocytes (CD19+) among styrene-exposed workers. The exposed workers showed a higher proportion of activation markers, namely DR and interleukin-2 receptors (CD25). Immunoglobulin subclasses were comparable in the two groups. An increased prevalence of abnormally low values was apparent for CD2+, CD3+, CD4+, CD4+45+ and CD11b subsets among workers exposed to styrene, whereas CD19+, DR+ and CD25+ showed an increased prevalence of abnormally high values. Natural killer-related phenotypes (CD56+, CD56+16+, and CD56+16-) were more expressed among styrene workers, with average increase of 30%. However, the frequency distribution of the lytic activity of natural killer cells against K-562 target cells was shifted towards lower values in the exposed workers as compared to control subjects. Dose-response relationships between indices of internal dose and prevalence of abnormal values were detectable for T lymphocyte subsets, NK phenotypes, and activation markers. These findings suggest that moderate exposure to styrene is associated with an altered distribution of lymphocyte subsets.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Quality assurance for immunochemical methods.

Recently developed immunochemical methods offer many advantages, although they may suffer from problems in standardisation due to the difficulties in the characterisation of immunological reagents. The reliability of the results is influenced by the availability of reference materials, calibrators, reagent kits and instruments. Since inter-laboratory quality control programmes have been limited by the lack of standard reference materials, laboratories using immunoassays should, at least, implement an internal quality control programme aimed at avoiding systematic errors, and adhere to the rules for good laboratory practice. The exchange of home-made materials as well as the control over pre-analytical factors (selection, collection and storage of specimens) within collaborative studies could be useful to the harmonisation of the measurement procedures. This paper deals with whether such quality requirements are or can be fulfilled with regard to early markers of nephrotoxic effects based on the immunochemical determination of proteins and kidney-derived antigens in urine or serum.

Humans

The assay of laminin fragments in serum and urine as an indicator of renal damage induced by toxins.

An ELISA procedure for the assay of laminin fragments in serum and urine is described. Samples from solvent-exposed workers and diabetic patients were studied. In cohorts exposed to perchloroethylene serum and urine laminin fragments were elevated but the urinary N-acetyl-beta-D-glucosaminidase (NAG) was unaffected. The data indicate that the urinary assay may be more specific for renal damage than the serum method. Differences in the excretion of NAG and urinary laminin fragments were observed in the diabetic groups suggesting that the excretion of these two components reflects different stages of severity of the disease.

Acetylglucosaminidase

Peripheral markers of catecholamine metabolism among workers occupationally exposed to manganese (Mn).

In a preliminary study of 11 men randomly selected in a ferro-alloy plant and of 15 control subjects, platelet monoamine oxidase (MAO) and serum dopamine beta-hydroxylase (DBH) activities were measured. A tendency towards lower MAO-B activity in the exposed workers as compared to control subjects (t = 1.95; P = 0.06) was found whereas DBH activity was similar. In the exposed group, a dose-effect relationship was noted between a manganese (Mn) cumulative exposure index (CEI) and DBH activity (R2 = 0.40, P < 0.05). Since DBH is an expression of catecholamine release, the relative increase in such activity could be envisaged as a compensatory mechanism to a reduced turnover rate as reflected by MAO-B activity. Owing to the limited sample size, these findings should be confirmed by further epidemiological and experimental studies.

Adult

Microdialysis as a tool to assess interstitial norepinephrine levels in adipose tissue of spontaneously hypertensive rats.

The microdialysis technique was applied to the study of norepinephrine (NE) metabolism in white adipose tissue of spontaneously hypertensive (SHR, n = 6) and normotensive Wistar-Kyoto (WKY, n = 6) rats. Mean concentrations of interstitial NE were much higher in SHR as compared to WKY (mean +/- SEM: 980.9 +/- 125.6 pg/ml vs 520.7 +/- 96.1 pg/ml; p = 0.01) over the 180 min experimental period. These results are consistent with the hypothesis that sustained outflow from nerve endings of the peripheral sympathetic system may play a role in the maintenance of arterial hypertension. Owing to its low invasiveness, the microdialysis technique allows to continuously monitor NE extracellular levels in conscious and freely-moving animals.

Adipose Tissue

Does occupational cobalt exposure determine early renal changes?

A cohort of workers occupationally exposed to cobalt (Co) dusts was examined to assess possible subclinical renal effects attributable to Co. Cross-sectional investigations involved 26 workers with a mean age of 34.2 (S.D., 8.3), chronically-exposed (median, 3.5 years; range, 0.9-11) to Co dusts in hard-metal manufacturing factories. Thirty-five healthy control workers, with a mean age of 32.4 (S.D., 4.6) were also examined. Individual interviews were used to exclude subjects with renal or systemic diseases, intake of nephrotoxic drugs, and exposure to known nephrotoxins. Exposure levels, assessed by ambient and biological monitoring, showed an estimated exposure approaching the ACGIH-recommended TLV of 50 micrograms/m3. Immunochemical methods were used to measure urinary albumin, retinol-binding protein (RBP), beta 2-microglobulin (beta 2m), and tubular brush-border antigens. The prevalence of abnormal values for early markers of renal dysfunction was similar in Co-exposed workers and in controls. However, within the reference interval, the cumulated frequency distribution for beta 2m was shifted towards higher values in the exposed group. No relationship was detected between renal markers and either intensity or duration of exposure. In spite of a limited number of observations, these findings suggest that the kidney is not a target organ during occupational exposure to Co.

Adult

Protracted high-dose interferon gamma therapy for chronic experimental nephropathy.

This study focused on the utility of interferon gamma (IFN gamma) as an anti-fibrotic drug in renal experimental fibrosis; the nephropathy was induced by two doses of Adriamycin (ADR) in 20 Sprague Dawley rats, 10 of which were randomly assigned to receive IFN gamma (45,000 UI) on alternate day for 16 weeks. At the end of the follow up, ADR rats treated with IFN gamma developed massive proteinuria, slight renal insufficiency, and presented diffuse glomerulosclerosis, tubulo interstitial infiltration and fibrosis. No difference was found in the composition of tubulo-interstitial infiltrates, mainly consisting in CD4+T lymphocytes with a minor component of CD8+T cells, in comparison with rats treated with ADR alone. These observations demonstrate the inefficacy of a protracted high-dose treatment with IFN gamma in chronic experimental nephropathy with interstitial fibrosis.

Animals

Renal effects of nifedipine and captopril in patients with essential hypertension and reduced renal reserve.

In this study we investigated the short-term effects of calcium channel blockers and angiotensin-converting enzyme inhibitors on renal hemodynamics and the urinary excretion of proteins with different relative mass in subjects with mild to moderate essential hypertension and apparently normal glomerular filtration rate but reduced renal functional reserve. Sixteen subjects underwent the following four treatments: (1) low-protein meal (0.2 g protein/kg body wt), (2) high-protein meal (1.3 g protein/kg body wt), (3) high-protein meal plus oral nifedipine (20 mg), and (4) high-protein meal plus oral captopril (50 mg). Two urine samples were obtained after meals. Blood samples were drawn at the midpoint of each 120-minute urine collection period. Urine and serum were tested for total protein, immunoglobulin G, albumin, alpha 1-microglobulin, retinol binding protein, and beta 2-microglobulin. Glomerular filtration rate and renal plasma flow were assessed by iothalamate and p-aminohippuric clearance, respectively. Compared with the high-protein meal alone, nifedipine elicited a clear-cut increase in the urinary excretion of total protein (+60%, P < .01), immunoglobulin G (+58%, P < .01), albumin (+25%, P < .05), retinol binding protein (+47%, P < .05), and beta 2-microglobulin (+52%, P < .05); captopril decreased the urinary excretion rate of immunoglobulin G (-26%, P < .05), albumin (-22%, P < .05), and beta 2-microglobulin (-34%, P < .05). The ratio between the clearances of immunoglobulin G and albumin was higher after nifedipine (+21%, P < .01) and unchanged after captopril (-9%, P = NS) compared with the high-protein meal alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Multiple interferences on catecholamine metabolism by tetrahydroisoquinolines (TIQs).

TIQs are thought to be formed by condensation between dopamine and certain metabolites of ethanol, organic solvents and anesthetic gases. Described here are experiments aimed at evaluating TIQs interference with catecholamine synthesis. Rat adrenal pheochromocytoma (PC12) cell lysates were exposed to benzyl-TIQ and phenyl-TIQ. The activities of tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) were measured by HPLC-based methods following exposure to variable concentrations of TIQs. The effects of TIQs on DBH activity were also assessed in human serum. Dixon plot analyses revealed that TIQs act on TH as competitive inhibitors with different affinity. Ki for benzyl- and phenyl-TIQ were 5 and 3 microM respectively. DBH activity in serum exposed to benzyl- and phenyl-TIQ ranging from 0.2 to 20 microM rose respectively by 12.5% to 58% for benzyl- and by 7.8% to 26% for phenyl-TIQ. Such TIQs interferences with catecholamine metabolism seem to account for dopamine (DA) depletion observed in parallel in vitro experiments on PC12 cells. The dose-dependent inhibition of TH and the increased activity of DBH together with the relatively low effective doses of TIQs suggest this mechanism as a possible explanation of the selective toxicity of styrene and other solvents to dopaminergic systems observed in rabbits following experimental exposure and suspected to occur in occupationally-exposed workers.

Animals

A case of asymptomatic giant aneurysm of the common hepatic artery.

The authors are presenting a case of asymptomatic giant aneurysm of the common hepatic artery in a 72 year-old woman. The patient came under our observation with a complete, recent diagnosis. However, two years earlier in another clinic, a para-hilar hepatic mass with partially calcified walls was incidentally found during an abdominal ultrasound. An abdominal TC without contrast medium showed the mass to be hepatic hydatid cysts. TC images after 2 years showed a growth in diameter from 7 to 12 cm. Having undergone an hysterectomy and a rectal prosthesis, technical difficulties occurred because of the aneurysm's characteristics. The authors would like to emphasize that the rarity of the lesion must not exclude it from the diagnostic protocol of abdominal masses: ultrasound must be accompanied by a Doppler and the TC must be made using a contrast medium. Furthermore, revascularization surgery may create difficulties that cannot be previously prevented.

Aged

Significance of albuminuria in the follow-up of acute poststreptococcal glomerulonephritis.

The present study was aimed at assessing the diagnostic value of urinary albumin (uA) excretion rate in the long-term follow-up of patients suffering from acute post-streptococcal glomerulonephritis (APSGN). 26 patients, who had presented primarily with nephritic syndrome and showing increased uA without a concomitant rise in total proteinuria (uTP) were followed-up for 131 months on average (range 36-288). At the last check, 14 patients did not show urinary abnormalities, 9 had a persistent increase in uA, 1 increased uTP and 2 renal insufficiency. Urinary and clinical signs of the disease were not seen during observation periods prolonged for 79 months on average (range 20-156) after normalization of uA. No pathological findings were remarked in biopsy specimens obtained in 3 patients when uA was normalized; in contrast, when both uTP and uA (12 cases) or when isolated uA (14 cases) were increased a pattern of diffuse mesangial proliferative glomerulonephritis was constantly observed. These results indicate that the abnormal uA excretion rate during long-term follow-up of APSGN allows to identify a subset of patients with persistent renal disease; conversely, the occurrence of normal uA seems to point to a good diagnostic and prognostic significance.

Acute Disease

[Primary aorto-enteric communication].

BACKGROUND: Primary aortoenteric communications are a rare and severe complication of abdominal aortic aneurysms or erosions by neoplastic diseases. Early diagnosis and surgical treatment are crucial. Postoperative morbidity and mortality are high. Diagnosis if often made intraoperatively, because the typical clinical feature (digestive haemorrhage, abdominal aneurysmal mass, abdominal pain) is often incomplete and critically ill patients require quick surgical decision and do not allow the use of sophisticated diagnostic tools. MATERIAL AND METHODS: Eight cases, observed through 13 years, are presented: six males and two females. Mean age was 61 years; male to female ratio was 3:1. In four cases (50%) a herald bleeding occurred during the days preceding hospital admission. Presenting symptoms were haematemesis (63%), melaena (87%), abdominal pain (63%); six subjects (75%) presented hemorrhagic shock. Only three patients (37%) were aware to be affected with abdominal aortic aneurysm before admission. Diagnosis was always made by clinical feature and urgent laparotomy: two preoperative duodenoscopies were not diagnostic. Aortoduodenal communication occurred in six cases: one of these was secondary to erosion of the aorta by a carcinoma of the pancreas. Aortogastric communication occurred once; the remainder case was a communication between a hypogastric artery aneurysm and the last ileal loop. RESULTS: Surgical operation was carried out in emergency in seven patients: the eight (tumour of the pancreas and aortoduodenal erosion) died before operation. Enteric defect repair, aneurysmectomy and aortic grafting was performed in six cases. In the last case (hypogastric-ileal communication) ligation of the hypogastric artery and ileal segmental resection was performed. Thirty days operative mortality was 58%. CONCLUSION: Despite early recognition and operation, primary aorto-enteric communication remains a severe life threatening disease, bringing high mortality rates. These are clearly affected by shock condition, but a correct surgical technique and prolonged postoperative antibiotic medication to avoid graft infection are mandatory to minimize mortality.

Adult

On the need of a sampling strategy in biological monitoring: the example of hexane exposure.

Ambient and biological monitoring of hexane exposure were repeatedly carried out in 14 female shoe makers. Airborne hexane (Ci-H) was measured in 4-h samples collected by a diffusive method. Urinary spot samples were collected before, during (at noon), and at the end of a work shift. 2,5-Hexanedione (2,5HD) in urine collected at noon was poorly related to morning Ci-H. End-of-shift 2,5HD were also poorly related to afternoon air samples. The correlation was still relatively low when end-of-shift 2,5HD was related to 8-h TWA Ci-H (r = 0.44; P < 0.01 on a linear scale, and r = 0.58, P < 0.01 on a log-log scale). End-of-shift 2,5HD levels estimated on the basis of pre-shift values using a mathematical model were much higher (2.3 times on average) than those experimentally measured during the study period. Owing to its relatively long half-time, 2,5HD seems to be influenced not only by current exposure, but also by hexane absorbed during the day(s) preceding sampling. The lack of a sampling strategy may account not only for inconsistencies between environmental and biological data, but also for a possible misuse of biological monitoring when utilized for risk assessment. Despite sometimes poor correlations with Ci-H, 2,5HD may still be preferred to other indicators as a marker of effective internal dose. A sampling strategy should ensure that measured values are representative of the individual risk for adverse effects.

Adult

Progression of chronic adriamycin nephropathy in leukopenic rats.

In this study, we examined the progression of chronic Adriamycin (ADR) nephropathy in mild leukopenic rats and tried to define the possible relationship between tubulointerstitial lesions and proteinuria in this model. Chronic ADR nephropathy was induced by 2 doses of ADR (2 mg/kg) in 32 Sprague-Dawley rats. Eight of these were randomly assigned to cyclophosphamide treatment (50 mg/kg), given intravenously every week, to keep the blood leukocyte count constantly lower than 5,000/mm3. Serial parameters were followed for 16 weeks including clearance studies with iothalamate and p-aminohippurate and the analysis of urinary protein composition by: (a) an enzymatic assay for beta-glucosidase; (b) specific ELISA using antibodies against rat albumin and RBP, and finally (c) two-dimensional electrophoresis. ADR-treated rats rapidly (within 2 weeks) developed massive proteinuria which was in constant increment throughout the disease evolution in each single component (i.e., high and low molecular weight proteinuria, enzymuria) and developed renal insufficiency. At week 8, in ADR rats, glomerulosclerosis was mild whereas tubulointerstitial infiltrates predominated, characterized mainly by CD4+ T lymphocytes while CD8+ T lymphocytes were inconspicuous, and macrophages were only occasionally present. All such alterations had worsened after 16 weeks when the tubulointerstitial infiltration was associated with marked interstitial fibrosis and tubular atrophy. Leukopenia induced by cyclophosphamide was in all cases associated with a net amelioration of renal histopathology reducing tubulointerstitial infiltrates (by 40%) and glomerulosclerosis (33 +/- 5 vs. 52.2 +/- 7.5% sclerotic glomeruli) and also ameliorated glomerular filtration indexes (Cl 780 +/- 40 vs. 447 +/- 66 microliters/min/kg-1). In spite of these differences, albuminuria and urinary-retinol-binding protein were comparable at weeks 4, 8 and 16 in this group, while urinary beta-glucosidase was decreased at week 16 (p < 0.001) in cyclophosphamide-treated rats. No other qualitative changes in urinary proteins were detectable by 2-dimensional electrophoresis during the disease development. We concluded that chronic leukopenia prevents interstitial cellular infiltration by lymphocytes, interstitial fibrosis and slows down the decline of renal function typical of chronic ADR nephropathy. Glomerulosclerosis is also reduced in leukopenic rats without any appreciable changes in the urinary excretion of high molecular weight proteins deriving from the glomerulus. Finally, the improvement in tubulointerstitial alteration is associated with the reduction in urinary lysosomal enzymes. Tubulointerstitial alterations are implicated with a prominent role in the progression towards renal failure in chronic ADR glomerulopathy.

Animals