Lysosomal exoglycosidases in serum and urine of patients with pancreatic adenocarcinoma.

Lysosomal exoglycosidases: N-acetyl-ß-D-hexosaminidase (HEX), ß-D-galactosidase (GAL), α-L-fucosidase (FUC) and α-D-mannosidase (MAN) modify oligosaccharide chains of glycoconjugates in endoplasmatic reticulum and/or Golgi apparatus and degrade them in lysosomes. In acid environment of lysosome, exoglycosidases degrade oligosaccharide chains of glycoproteins, glycolipids and glycosaminoglycans by eliminating single sugars from the edges of oligosaccharide chains. Neoplasms change biochemical processes in tissues and may significantly change the activity of many enzymes including the activity of lysosomal exoglycosidasses in serum and urine of persons with neoplasmatic diseases. The aim of the present paper was evaluation the activity of HEX, GAL, FUC and MAN in serum and urine of patients with pancreatic adenocarcinoma. Serum and urine samples were collected from 15 patients with adenocarcinoma of the pancreas and 15 healthy persons. The activity of lysosomal exoglycosidases was determined by the method of Marciniak et al. adapted to serum and urine of patients with adenocarcinoma of the pancreas. Our results indicate significant decrease in activity of GAL (p=0.037) in serum of patients with pancreatic adenocarcinoma, significant increase in activity of HEX (p<0.001) and FUC (p=0.027) in serum, and HEX (p=0.003) in urine, as well as significant decrease of FUC (p=0.016) and MAN (p=0.029) in urine o patients with adenocarcinoma of the pancreas, in comparison to the control group. Increase in activity of some lysosomal enzymes in serum and urine of pancreatic adenocarcinoma patients, may indicate on destruction of pancreatic tissue by pancreatic adenocarcinoma. Determination of the HEX, GAL, FUC and MAN in serum and urine may be useful in diagnostics of pancreatic adenocarcinoma.

N-acetyl-beta-D-hexosaminidase and its isoenzymes A and B in blood serum and urine, as a potential colon cancer markers.

BACKGROUND/AIMS: Evaluation of N-acetyl-beta-D-hexosaminidase (HEX), and its isoenzymes A (HEX A) and B (HEX B) activity in blood serum and urine as potential markers of colorectal cancer. METHODOLOGY: The study was performed in blood serum and urine of 32 patients with adenocarcinoma, 6 with adenocarcinoma mucinosum of the colon, and 20 healthy people. The activity of HEX, HEX A and HEX B was determined in blood serum and urine by spectrophotometric method of Marciniak et al. The concentration of CEA was determined in blood serum by immunoenzymatic method (MEIA). The concentration of protein was assessed by the Lowry method, whereas the concentration of creatinine in urine by the Jaffe method (without deproteinization). RESULTS: A significant increase in the concentration of HEX, HEX A and HEX B activity was proved in serum and urine of patients with colon adenocarcinoma. In patients with colon adenocarcinoma mucinosum, the higher activity of HEX was revealed in blood serum compared to healthy people, and the significantly higher activity of HEX and HEX B expressed as pKat/mg of creatinine, was found in urine. We observe a significant increase in the activity of HEX, HEX A and HEX B expressed in pKat/mg of creatinine was found in urine of patients bearing tumor of diameter 6.0-7.0 cm in comparison to patients with tumor of diameter 4.0-5.0 cm. CONCLUSIONS: The present study results suggest that determination of HEX, HEX A and HEX B activity in blood serum and urine may be used to detect colon cancer in its early stages. However, the use of HEX, HEX A and HEX B activity in oncological diagnostics requires further studies on a larger group of patients.

Effect of smoking on activity of N-acetyl-beta-hexosaminidase in serum and urine of renal cancer patients.

OBJECTIVES: To compare N-acetyl-beta-hexosaminidase (HEX) activity in the serum and urine of smokers as well as non-smokers with renal cancer, and healthy people. DESIGN AND METHODS: To assess hexosaminidase activity the level of p-nitrophenol released from p-nitrophenol derivatives was measured. RESULTS: The activity of enzyme was significantly higher in cancer group, with the highest activity in non-smokers. CONCLUSIONS: Cigarette smoking can inhibit, by the influence on HEX activity, catabolism of oligosaccharide chains in cancer tissues.

[N-acetyl-beta-hexosaminidase--marker of damage to renal proximal tubules]. / N-acetylo-beta-heksozoaminidaza--marker uszkodzenia cewek nerkowych blizszych.

Cells of the renal epithelium synthesize and excrete to urine many enzymes. Among more than 50 enzymes produced by epithelial cells of proximal tubules, only few have a diagnostic value. Determination of the enzymatic activities in urine is sensitive and not invasive method for evaluation the function of renal tubules. Urinary N-acetyl-beta-hexosaminidase (HEX) activity is approved and practically utilized marker of the renal function. HEX is a lysosomal exoglycosidase taking part in catabolism of the sugar chains of glycoconjugates (glycoproteins, glycolipids and proteoglycans). HEX catalyses release of N-acetylglucosamine and N-acetylgalactosamine from a non reducing ends of glycoconjugates. In urine of healthy persons activity of HEX is negligible, but significantly increases after damage to the proximal tubules. The cells of renal proximal tubules are very sensitive to hypoxia. Therefore all renal processes with hypoxia lead to dysfunction of proximal renal tubules and release HEX to urine. Increased activity of HEX in urine was found after intoxication by heavy metals, nephrotoxic drugs, contrast media, fewer, bacterial as well as immunological nephritis and hypertension, diabetes, neoplasms and during renal graft rejection. In the paper we presented review of literature concerning HEX, and its presence in renal tissue and urine, as well as application in diagnostics.

Renal tubular dysfunction in schizophrenic patients treated with antipsychotic drugs.

Several factors have been considered in relation to the free radical formation in schizophrenia, such as the disease itself, drug treatment and smoking. Several chemicals and drugs may cause damage to the renal tubules by different subcellular mechanisms including oxidative stress, and the aim of our study was the investigation of tubular dysfunction in schizophrenic patients. The urinary excretion of beta-N-acetylhexosaminidase (Hex) and its isoenzymes Hex A and Hex B, alpha1-microglobulin, albumin, total proteins and fractionated porphyrins were determined in 45 schizophrenic patients treated with first- and second-generation antipsychotics. In 7 patients, an increase in proteinuria of tubular origin was found, and in one as a result of mixed glomerular/tubular origin. The group of patients had a significantly higher level of excretion than the control group (n = 54) of total Hex (p < 0.001), Hex A (p < 0.05), Hex B (p < 0.001) and the relative proportion of this isoenzyme (p < 0.001). In some cases with normal levels of total Hex and urinary alpha1-microglobulin, the proportion of Hex B was already increased. Significant correlations were found for total Hex and its isoenzymes with alpha1-microglobulin (p < 0.001). Also, the porphyrins had significant correlations with total Hex (p < 0.001), Hex A (p < 0.05), Hex B (p < 0.005) and alpha1-microglobulin (p < 0.001). In the group of patients studied, it was possible to reveal early tubular cell damage (affected structural integrity) with increased excretion of Hex B, possibly mediated by free radicals, previous to the decrease in tubular reabsorption of proteins with low molecular mass filtered by the glomerulus (affected functional integrity).

[Suspected case of pseudohypoparathyroidism type II].

We experienced a suspected case of pseudohypoparathyroidism type II. The patient came to our emergency room with no thermal convulsion. The Ellsworth-Howard test was applied to the patient to determine the type of PHP.

A comparative study of three kidney biomarker tests in autosomal-dominant polycystic kidney disease.

BACKGROUND: The relationship between the progress of tubular damage and renal insufficiency in autosomal-dominant polycystic kidney disease (ADPKD) is a subject of doubtless interest, and is the object of this present work. METHODS: A total of 92 adult ADPKD patients of both genders were studied, none of which presented end-stage renal disease (ESRD), and classified according to an ultrasound score based on kidney size and number of cysts. Urinary albumin and beta-N-acetylhexosaminidase (Hex) and its isoenzymes were determined, together with serum glutathione peroxidase, cystatin C, creatinine, and urea. RESULTS: A frequent elevation of the urinary Hex was found and an alteration of its isoenzymatic profile, with 31% of the normotensive patients with normoalbuminuria already presenting an increased proportion of Hex B isoenzyme. Keeping age constant, a partial significant correlation was found between the ultrasound score and the proportion of Hex B (r = 0.352, P < 0.05), but not with albuminuria or cystatin C. In 42 patients the different biochemical variables were again determined after 1 year, finding that in the 13 normotensive patients with normoalbuminuria there had been a significant decrease in the concentration of cystatin C (P < 0.05), and a significant increase in the urinary excretion of albumin and Hex B isoenzyme (P < 0.05). By the other hand, in the other 29 patients with micro- or macroalbuminuria and hypertension, no significant differences were found. CONCLUSION: The results point toward an important participation of tubular damage in the pathogenesis of this disease. It may also be suggested that in normotensive and normoalbuminuric ADPKD patients, a gradual increase of glomerular filtration would be produced. After the start of hypertension and microalbuminuria, the glomerular filtration rate (GFR) would decrease progressively, although more slowly.

Sialylation analysis of O-glycosylated sialylated peptides from urine of patients suffering from Schindler's disease by Fourier transform ion cyclotron resonance mass spectrometry and sustained off-resonance irradiation collision-induced dissociation.

A strategy based on Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) for screening of complex glycoconjugate mixtures containing O-linked glycopeptides and O-glycosylated amino acids with alpha-N-acetylgalactosaminyl residues is presented. To detect and identify O-glycoforms present in urine of patients suffering from hereditary N-acetylhexosaminidase deficiency (known as Schindler's disease), present at 100 times higher concentrations than in urine of healthy controls, new accurate methods for mapping and sequencing were required. In the mass spectrometric analysis particular attention has to be paid to original sialylation patterns, because of the potential lability of the sialic acid moiety during the desorption/ionization process. Negative ion nanoelectrospray ionization (nanoESI) FTICR-MS at 9.4 T is shown here to represent a method of choice for identification of single components in such complex glycomixtures due to high resolution and mass accuracy. By optimization of sustained off-resonance irradiation collision-induced dissociation tandem mass spectrometry (SORI-CID-MS(2)) in the negative ion mode, the type and sequence of the sialylated glycopeptide components were determined from their fragmentation patterns. Additionally, implementation of SORI-CID-MS(3) provides detailed information for sialylation analysis. The potential diagnostic value of this approach is discussed.

Determinación de la ß-N-acetilhexosaminidasa en orina utilizando un método espectrofotométrico continuo automatizado / Determination of ß-N-acetylhexaminidase in urine using an automated continuous spectrophotometric method

Se adaptó a un analizador centrífugo Cobas Bio un método continuo para la determinación de la ß-N-acetilhexosaminidasa (Hex, EC 3.2.1.52) en orina utilizando 3,3'-diclorofenolsulfonftaleinil-N-acetil-ß-D-glucosaminido como sustrato. En el estudio de la precisión intraserie se obtuvieron unos coeficientes de variación de 0,77-2,10 por ciento y en el de precisión interserie de 0,89-2,83 por ciento. En todos los casos los coeficientes de variación fueron inferiores a los médicamente aceptables según criterios del Colegio de Patólogos Americanos. Se encontró una excelente correlación entre los resultados obtenidos con esta técnica y con un método discontinuo utilizando 2-metoxi-4-(2'-nitrovinil)-fenil-N-acetil-ß-D-glucosaminido como sustrato (n = 100, r = 0,997)> No se ha podido confirmar el hecho señalado por algunos autores, de que el efecto inhibidor de la urea sobre la actividad enzimática de la Hex sea mayor al aumentar el pH en que se lleva a cabo la hidrólisis de estos sustratos cromogénicos

Elevated urinary excretion of beta-hexosaminidase in smokers.

The excretion of urinary beta-hexosaminidase in 203 apparently healthy males and females correlated significantly with their tobacco consumption (r = 0.26; p less than 0.001). Those who smoked more than 10 g tobacco per day excreted significantly more beta-hexosaminidase in urine than non-smokers, which indicates the presence of a low grade renal injury in smokers.

Effect of acid-base changes on urinary hydrolases in Fabry's disease after renal transplantation.

Fabry's disease, which is characterized by alpha-galactosidase A (AG) deficiency, causes early renal failure. Kidney transplants do not reliably supply the deficient enzyme. To assess both urinary excretion of AG by the transplant and the relationship between urine and serum hydrolase activity, acute and chronic acid-base studies were performed in normal control subjects and in the patient with Fabry's disease who had undergone renal transplantation. For the acute studies, alkalosis was induced by intravenous infusion of sodium bicarbonate and acidosis was induced by ingestion of ammonium chloride. The chronic study involved long-term ingestion of NH4Cl by only the patient with Fabry's disease. The results show that AG is secreted by the renal graft. Urinary hydrolase excretion was increased by acute alkalinization and decreased by acute acidification. Acute, but not chronic, acidification increased the patient's serum AG activity, indicating that long-term acidification is not useful for treating Fabry's disease after transplantation. The large changes in hydrolyase excretion induced by acute and chronic acid-base changes show the difficulty of using lysosomal enzymuria as a diagnostic marker for renal disorders without knowledge of acid-base conditions.

Multiple marker validity of urinary hexosaminidase and polyamines in haematopoietic malignancy.

Proliferation end products are of considerable value as tumour markers. Hexosaminidase and polyamines were significantly elevated in urine of patients with leukemia and lymphoma (p less than 0.001). Interfering low molecular weight compounds were eliminated by a microcolumn centrifuge method. Urinary polyamines were estimated by high voltage electrophoresis technique. Marked increase in these markers in untreated patients and subsequent decrease on effective chemotherapy support the use of these two tumour markers to monitor therapeutic response.