Altered regulation of serum lysosomal acid hydrolase activities in Parkinson's disease: A potential peripheral biomarker?

INTRODUCTION: Recent studies have indicated that lysosomal dysfunction contributes to the development of idiopathic Parkinson's disease (PD). It is uncertain whether dysregulation of serum lysosomal acid hydrolase activity exists in sporadic PD patients compared with normal controls (NCs) and parkinsonian syndrome (PS) patients. METHODS: Sporadic PD patients without GBA1 mutations (n = 68) were matched with normal controls (n = 45), and parkinsonian syndrome patients (n = 32) in terms of family history, age, and sex. We measured the activities of lysosomal enzymes, α-galactosidase, ß-galactosidase, and ß-hexosaminidase and examined the possible correlations between lysosomal acid hydrolase activities with age in NCs, PD, and PS patients. RESULTS: ß-Galactosidase activity was significantly higher in the PD and PS than in the NC group (P < 0.001). The ß-galactosidase to α-galactosidase and ß-hexosaminidase to ß-galactosidase activity ratios were more useful for distinguishing PD and PS patients from NCs (P < 0.0001). Furthermore, α-galactosidase activity was significantly higher in PS patients than both PD and NC groups (p = 0.04). ß-Galactosidase and α-galactosidase activities exhibited a statistically significant negative correlation with age in NCs, and ß-hexosaminidase activity showed a positive correlation with age in PS. However, PD patients did not show any of these correlations. CONCLUSION: Our results suggest the presence of an unknown regulatory mechanism(s) of serum acid hydrolase activities with aging in the normal population and abnormalities in their regulation in PD and PS patients. However, the pattern of dysregulation in these two groups is different. Thus, serum lysosomal acid hydrolase activity can be used as a peripheral biomarker for PD.

Positive Correlation of Serum N-Acetyl-ß-hexosaminidase with Markers of Atherosclerosis in Diabetes Type 2 Patients with Mild Symptoms of Depression and Anxiety.

BACKGROUND: Analysis of the correlation between diabetes type 2 (DT2) and serum N-acetyl-ß-hexosaminidase (HEX) activity with parameters of fat metabolism and symptoms of anxiety and depression. MATERIAL AND METHOD: The study was performed using a random sample of 40 DT2 patients (22 women and 18 men) between the ages of 43 and 71 (median 59) and 40 control persons (28 women and 12 men) between the ages of 18 and 64 (median 46). The activity of HEX was determined by a colorimetric method. The activity of the serum exoglycosidase was expressed in pkat/mL. Each participant underwent Hamilton tests, to evaluate level of anxiety and depression. Additionally, the HEX activity and concentration of particular lipidograms were monitored using a blood sample from each participant. RESULTS: In DT2 patients, a significant positive correlation was found between serum HEX activity and the concentration of serum cholesterol LDL fractions, triacylglycerols (TAG), and Castelligro atherogenic indexes. A significantly increased level of anxiety and depression in comparison to the control group was found as well. CONCLUSION: Serum HEX activity in DT2 patients is a better marker of atherosclerosis than serum total cholesterol level in persons with mild symptoms of depression and anxiety. In DT2 patients, a routine testing of anxiety and depression is recommended. Early detection of these disorders creates the possibility for treatment, an improvement in a patient's quality of life, and the overall longevity of DT2 patients.

Reference Data on Neonatal Serum N-Acetyl-ß-hexosaminidase Activity.

BACKGROUND: Determination of neonate serum's N-acetyl-ß-hexosaminidase (HEX) activity and correlation results with Apgar scale and factors routinely determined in newborn serum. AIMS: Providing reference values of neonates serum HEX activities, and indicate their diagnostic significance. STUDY DESIGN: The study was performed using random serum samples of 111 infants (53 ♂/58 ♀), aged 1-30 days. The activity of HEX was determined colorimetrically and expressed in nKat/L. RESULTS: Serum HEX activity of 111 newborns was 360.5 ± 114.0 nKat/L and significantly positively correlated with gestation week at the day of delivery, birth weight, weight on day of blood collection, sex, and serum CRP. CONCLUSIONS: Reference values presented for neonatal serum activities of HEX may be used in neonatal diagnostics, for example, to detect inflammation and other diseases or for early assessment of the risk of Tay-Sachs and Sandhoff diseases.

Determination of N-acetyl-ß-hexosaminidase in serum from hemolyzed blood.

BACKGROUND: Determination of lysosomal N-acetyl-ß-hexosaminidase (HEX) in serum from hemolyzed blood, creates serious analytical problems, because hemoglobin absorbs light at a similar wavelength like 4-nitrophenol, which is released from artificial substrate. OBJECTIVE: The objective of the work was to adapt a manual method to allow analysis of HEX in hemolyzed samples. METHODS: Serums without and with hemolysis were incubated with 4-nitrophenol-N-acetylglucosamine as a substrate. Released 4-nitrophenol was determined colorimetrically. After the incubation of the serum from hemolyzed blood with substrate, hemoglobin was precipitated with trichloroacetic acid (TCA) before 4-nitrophenol determination. RESULTS: The mean concentration of HEX activity in non-hemolyzed and hemolyzed blood of the same patients, determined with non-modified and modified methods had no significant differences, and they are: 243.12±119.76 and 233.99±108.76pkat/mL, respectively. A coefficient of correlation between non-modified and modified methods equals the 0.98. For HEX determination with the modified method in serum from hemolyzed blood, optimal reaction time was 60min, pH of reaction mixture was 4.7, and Km was 0.11mMm. CONCLUSION: HEX determinations in the same serums from non-hemolyzed blood by the non-modified method and hemolyzed blood with the modified method, gave similar results with a 0.98 coefficient of correlation. The modified method is appropriate for HEX determination in serum from hemolyzed blood.

Leukocyte glucocerebrosidase and ß-hexosaminidase activity in sporadic and genetic Parkinson disease.

BACKGROUND: Recent reports have shown that the activities of lysosomal enzymes are altered in the CNS of sporadic PD (sPD) without GBA mutations. We hypothesized that the activities of lysosomal enzymes are altered in peripheral blood leukocytes (PBLs) of patients with sPD and other genetic parkinsonism. METHODS: Glucocerebrosidase and ß-hexosaminidase activities in PBLs were measured in 36 patients with sPD, 5 PD patients with PARK2 mutations, 10 patients with spinocerebellar ataxia (SCA) 17 with parkinsonism, and 20 healthy controls. RESULTS: The glucocerebrosidase and ß-hexosaminidase activities were not different in patients with sPD, PD with PARK2 mutations, and SCA17 with parkinsonism from those of the controls. In the patients with sPD, the activity of GCase was positively correlated with disease duration. CONCLUSION: The glucocerebrosidase and ß-hexosaminidase activities in PBLs cannot be used as a biomarker in sPD and other genetic parkinsonism.

Propofol Attenuates Small Intestinal Ischemia Reperfusion Injury through Inhibiting NADPH Oxidase Mediated Mast Cell Activation.

Both oxidative stress and mast cell (MC) degranulation participate in the process of small intestinal ischemia reperfusion (IIR) injury, and oxidative stress induces MC degranulation. Propofol, an anesthetic with antioxidant property, can attenuate IIR injury. We postulated that propofol can protect against IIR injury by inhibiting oxidative stress subsequent from NADPH oxidase mediated MC activation. Cultured RBL-2H3 cells were pretreated with antioxidant N-acetylcysteine (NAC) or propofol and subjected to hydrogen peroxide (H2O2) stimulation without or with MC degranulator compound 48/80 (CP). H2O2 significantly increased cells degranulation, which was abolished by NAC or propofol. MC degranulation by CP further aggravated H2O2 induced cell degranulation of small intestinal epithelial cell, IEC-6 cells, stimulated by tryptase. Rats subjected to IIR showed significant increases in cellular injury and elevations of NADPH oxidase subunits p47(phox) and gp91(phox) protein expression, increases of the specific lipid peroxidation product 15-F2t-Isoprostane and interleukin-6, and reductions in superoxide dismutase activity with concomitant enhancements in tryptase and ß-hexosaminidase. MC degranulation by CP further aggravated IIR injury. And all these changes were attenuated by NAC or propofol pretreatment, which also abrogated CP-mediated exacerbation of IIR injury. It is concluded that pretreatment of propofol confers protection against IIR injury by suppressing NADPH oxidase mediated MC activation.

The interaction between oxidative stress and mast cell activation plays a role in acute lung injuries induced by intestinal ischemia-reperfusion.

BACKGROUND: Both oxidative stress and mast cells are involved in acute lung injuries (ALIs) that are induced by intestinal ischemia-reperfusion (IIR). The aim of this study was to further investigate the interaction between oxidative stress and mast cells during the process of IIR-induced ALI. MATERIALS AND METHODS: Thirty adult Sprague-Dawley rats were randomly divided into five groups: sham, IIR, IIR + compound 48/80 (CP), N-acetylcysteine (NAC) + IIR, and NAC + IIR + CP. All rats except those in the sham group were subjected to 75 min of superior mesenteric artery occlusion, followed by 2 h of reperfusion. The rats in the NAC + IIR and NAC + IIR + CP groups were injected intraperitoneally with NAC (0.5 g/kg) for three successive days before undergoing IIR. The rats in the IIR + CP and NAC + IIR + CP groups were treated with CP (0.75 mg/kg), which was administered intravenously 5 min before the reperfusion. At the end of the experiment, lung tissue was obtained for pathologic and biochemical assays. RESULTS: IIR resulted in ALI, which was detected by elevated pathology scores, a higher lung wet-to-dry ratio, and decreased expression of prosurfactant protein C (P < 0.05). Concomitant elevations were observed in the expression levels of the nicotinamide adenine dinucleotide phosphate oxidase subunits p47(phox) and gp91(phox) and the levels of hydrogen peroxide and malondialdehyde. However, superoxide dismutase activity in the lung was reduced (P < 0.05). The level of interleukin 6, the activity of myeloperoxidase, and the expression of intercellular adhesion molecule 1 were also increased in the lung. IIR led to pulmonary mast cell degranulation and increases in the plasma and pulmonary ß-hexosaminidase levels, mast cell counts, and tryptase expression in lung tissue. CP aggravated these conditions, altering the measurements further, whereas NAC attenuated the IIR-induced ALI and all biochemical changes (P < 0.05). However, CP abolished some of the protective effects of NAC. CONCLUSIONS: Oxidative stress and mast cells interact with each other and promote IIR-induced ALI.

The mechanism of sevoflurane preconditioning-induced protections against small intestinal ischemia reperfusion injury is independent of mast cell in rats.

The study aimed to investigate whether sevoflurane preconditioning can protect against small intestinal ischemia reperfusion (IIR) injury and to explore whether mast cell (MC) is involved in the protections provided by sevoflurane preconditioning. Sprague-Dawley rats exposed to sevoflurane or treated with MC stabilizer cromolyn sodium (CS) were subjected to 75-minute superior mesenteric artery occlusion followed by 2-hour reperfusion in the presence or absence of MC degranulator compound 48/80 (CP). Small intestinal ischemia reperfusion resulted in severe intestinal injury as demonstrated by significant elevations in intestinal injury scores and p47(phox) and gp91(phox), ICAM-1 protein expressions and malondialdehyde and IL-6 contents, and MPO activities as well as significant reductions in SOD activities, accompanied with concomitant increases in mast cell degranulation evidenced by significant increases in MC counts, tryptase expression, and ß-hexosaminidase concentrations, and those alterations were further upregulated in the presence of CP. Sevoflurane preconditioning dramatically attenuated the previous IIR-induced alterations except MC counts, tryptase, and ß-hexosaminidase which were significantly reduced by CS treatment. Furthermore, CP exacerbated IIR injury was abrogated by CS but not by sevoflurane preconditioning. The data collectively indicate that sevoflurane preconditioning confers protections against IIR injury, and MC is not involved in the protective process.

Microheterogeneity of serum ß-hexosaminidase in chronic alcohol abusers in a driver's license regranting program.

BACKGROUND: Carbohydrate-deficient transferrin (CDT) is one of the best indicators for chronic alcohol abuse and detection of relapse. In this study, we explore the microheterogeneity of ß-hexosaminidase (ß-HEX) in chronic alcohol abusers in the framework of a driver's license regranting program. Studies have shown that increased serum activity of ß-HEX B (isoforms P, I, and B) may be a sensitive marker for chronic alcohol abuse. Here, we describe methodology, limitations, and correlation of ß-HEX isoforms with CDT. METHODS: CDT was assayed at the central laboratory of the Ghent University Hospital by capillary zone electrophoresis, measured on the Capillarys 2™ system and was expressed as a percentage of total serum transferrin (%CDT). Serum of chronic alcohol abusers was compared to nonheavy drinkers using agarose gel isoelectric focusing (IEF). Total ß-HEX activity was assayed fluorimetrically following preparative IEF in 81 subjects. ß-HEX isoforms were investigated and compared between nonheavy drinkers and heavy drinkers. RESULTS: Agarose gel IEF shows additional cathodal bands in serum of chronic alcohol abusers. Mean total ß-HEX activity between pH 6.8 and 7.7, designated as HEX-7, showed the highest correlation with %CDT (r = 0.70, p < 0.0001, n = 68). In a selected subgroup, where CDT could not be quantified (n = 13) because of an atypical electropherogram, HEX-7 was in concordance with either estimated %CDT value or liver enzyme activities. CONCLUSIONS: In this proof-of-concept study, we introduce a novel approach to quantify ß-HEX isoforms using preparative IEF and fluorimetry. A highly significant correlation of HEX-7 and %CDT has been found. Because of exclusion of the P isoform, HEX-7 could be a useful supplementary marker for detecting chronic alcohol abuse.

Ym1, an eosinophilic chemotactic factor, participates in the brain inflammation induced by Angiostrongylus cantonensis in mice.

Angiostrongylus cantonensis is an emerging zoonotic pathogen that has caused hundreds of cases of human angiostrongyliasis worldwide. The larva in nonpermissive hosts cannot develop into an adult worm and can cause eosinophilic meningitis and ocular angiostrongyliasis. The mechanism of brain inflammation caused by the worm remains poorly defined. According to previous data of GeneChip, Ym1 in the brain of mice 21 days after infection with A. cantonensis was highly upregulated to over 7,300 times than the untreated mice. Ym1 is an eosinophilic chemotactic factor with the alternative names of chitinase-3-like protein 3, eosinophil chemotactic cytokine, and ECF-L. Ym1 displays chemotactic activity for T lymphocytes, bone marrow cells, and eosinophils and may favor inflammatory responses induced by parasitic infections and allergy. It has been reported that Ym1 is synthesized and secreted by activated macrophages during parasitic infection (Chang et al., J Biol Chem 276(20):17497-17506, 2001). In the brain, microglia are alternatively activated macrophage-derived cells which are the key immune cells in central nervous system inflammation. To explore the role of Ym1 in inflammation caused by A. cantonensis-infected mice, we examined the levels of Ym1 in the sera and cerebrospinal fluid (CSF) of the infected animals, followed by detection of the mRNA expression level of Ym1 in various organs including the brain, lung, liver, spleen, and kidney and of the cytokines IL-5 and IL-13 in the brain of the infected mice with or without intraperitoneal injection of minocycline (an inhibitor of microglial activation) by real-time reserve transcription PCR. Furthermore, immunolocalization of Ym1 in the brains of the infected mice was observed by using a fluorescence microscope. Our results showed that Ym1 was most highly expressed in the brains and CSF of the infected mice along with the process of inflammation. The antibody localized Ym1 to the microglia in the brain of the mice in both infection and minocycline + infection groups. And as in the brain, the mRNA level of Ym1 changed more obviously than IL-5 and IL-13. The study implies that Ym1 might serve as an alternative potential pathological marker which is detected not only in the sera and CSF but also in the brains of the infected mice and Ym1 secreted by microglia might be involved in eosinophilic meningitis and meningoencephalitis caused by A. cantonensis infection.

Mucolipidosis type III in an adolescent presenting with atypical facial features and skeletal deformities.

Mucolipidosis type III (MLIII) (MIM# 252600) is an uncommon autosomal recessive disorder that results from deficiency of the multimeric enzyme, UDP-N-acetylglucosamine-1-phosphotransferase. The enzymatic defect results in deficiencies of lysosomal degradative enzymes with concomitant intracellular accumulation of both partly degraded glycosaminoglycans and sphingolipids leading to clinical manifestations such as short stature, developmental delay and other structural abnormalities. The diagnosis is challenging since musculoskeletal presentation may mimic some of the rheumatic and metabolic disorders. We herein report on a 13-year-old adolescent who was admitted to our rheumatology clinic because of progressive joint stiffness and deformities of her hands. The clinical and radiological findings led us to the diagnosis of MLIII despite negative urinary aminoglycosyaminoglycans. Therefore we decided to check for the presence of elevated activities of alpha-mannosidase and beta-hexosaminidase A+B in the plasma which was actually the case and confirmed the clinical diagnosis ofMLIII.

A rapid and low-cost approach to evaluate the allergenicity of herbal injection using HPLC analysis.

Herbal medicines have ever been thought harmless, but it is obviously not true. Many adverse reports emerged with the development of their popular application in the world. Allergic reactions, especially serious immediate hypersensitivity, frequently occurred when herbal injections were used in clinic and made this ever prevailing agent nearly disappear in China. The aim of this study is to establish a rapid and economical method for the prediction of the allergenicity of herbal injections. Ovalbumin (OVA) and four other herbal injections, in which two of them were well known for their allergenicity, were selected to sensitize and stimulate the animals. Serotonin in the animal serum was detected with HPLC to reflect the anaphylactic response and compared with the other cytokines which could mediate the anaphylaxis, including histamine, IgE and ß-hexosaminidase. The results suggest that serotonin can be detected quickly and has good correlation with the other allergy-related cytokines. It is a promising way for predicting the allergenicity of the herbal injections and those complicated natural products.

Cholesteatoma-associated pathogenicity: potential role of lysosomal exoglycosidases.

UNLABELLED: The enzymatic profile of lysosomal exoglycosidases in middle ear cholesteatoma has not been well known. The assessment of glycoconjugate catabolism may contribute to a better understanding of cholesteatoma pathogenesis. OBJECTIVE: The study aim was to evaluate catabolic processes of glycoproteins, glycolipids, and proteoglycans in cholesteatoma through outlining the concentration of N-acetyl-ß-hexosaminidase (HEX), ß-glucuronidase (GLUC), and ß-galactosidase (GAL) activity as well as in serum of cholesteatoma patients and healthy volunteers. STUDY DESIGN: Acquired cholesteatomas (n = 25) and normal retroauricular skin specimens (n = 25) were taken during surgery as well as serum from cholesteatoma patients and healthy volunteers. HEX, GAL, and GLUC activity was assessed on basis of p-nitrophenol release from derivatives of the substrate (HEX: N-acetylglucosamine i N-acetylgalactosamine, GAL from galactose, and GLUC from glucuronide). RESULTS: The mean concentration of activity of HEX 1142.39 pKat/ml, GAL 8.90 pKat/ml, and GLUC 14.06 pKat/ml was significantly higher compared with the concentration of enzyme activity in normal tissue: HEX 267.65 pKat/ml, GAL 3.44 pKat/ml, and GLUC 3.90 pKat/ml. In the serum of cholesteatoma patients, the mean concentration of enzyme activities were as follows: HEX 641.62 pKat/ml, GAL 4.55 pKat/ml, and GLUC 12.80 pKat/ml and were significantly higher compared with the concentration of HEX activity (215.75 pKat/ml), GAL (1.89 pKat/ml), and GLUC (5.51 pKat/ml) in the serum of the healthy control group. In cholesteatoma compared with the normal tissue, there is an increase of the glycoconjugate catabolism due to significantly higher concentration of HEX, GAL, and GLUC activity in cholesteatoma. Cholesteatoma causes systemic reaction due to the increase of HEX, GAL, and GLUC activity in patient serum.

Effect of sample collection, temperature and time of storage on ß-galactosidase and total hexosaminidase activities in dried blood collected on filter paper.

BACKGROUND: Dried blood spots (DBS) on filter paper is a valuable sampling technique in clinical chemistry, but the stability of enzymes used in the diagnosis of lysosomal storage diseases (LSDs) needs to be evaluated. METHODS: In a first experiment, blood from 20 subjects was collected using a syringe without additives and distributed into EDTA tubes, heparin tubes, and spotted on filter paper for the comparison of sampling effects. In a second experiment, blood from 30 healthy subjects was spotted on filter paper and analyzed for ß-galactosidase and total hexosaminidase activities after storage of the samples at different temperatures for up to 180 days. RESULTS: Initially, we observed that enzyme activities were the same, independent of the collection method. When DBS was stored at 37°C the activity of ß-galactosidase dropped to 85% of the initial value after 180 days (p<0.05). At all other temperatures (-20°C, 4°C and 25°C), the results were within the methodological error. Total hexosaminidase activity did not change significantly during the entire study period and at different storage temperatures. CONCLUSIONS: The two enzymes investigated in the present study may be stored for up to 17 days (ß-galactosidase) or 180 days (total hexosaminidase) until analysis without loss of activity.

N-Acetyl-ß-D-hexosaminidase in gestational diabetes mellitus - a preliminary study.

PURPOSE: N-Acetyl-ß-D-hexosaminidase (HEX) is an exoglycosidase which has been extensively studied and which has been used as a marker for inflammation. It was therefore thought that measurement of the activity of this enzyme might be useful in diagnosing gestational diabetes mellitus (GDM) as this condition is frequently associated with inflammation. The main object of the study was the determination of N-acetyl-ß-D-hexosaminidase activity in women with GDM and 3 months postpartum in comparison with control groups of non-pregnant and healthy pregnant women. MATERIAL AND METHODS: Twenty-five blood serum samples from women with GDM and women 3 months postpartum; 20 blood serum samples from non-pregnant and healthy pregnant women (control groups) were enrolled into the study. Serum was prepared from all blood samples and HEX activity was measured by the method of Chateriee et al. (modified by Zwierz et al). RESULTS: A statistically significantly increase in the activity of HEX in the GDM blood serum was found as compared to the control groups (p<0.05). Further analysis showed a statistically significant decrease in the activity of HEX among postpartum women, but the level of enzyme activity was still above the normal control level in comparison to the control group of nonpregnant healthy women (p<0.05). CONCLUSIONS: Changes in the activity of HEX appear to be involved in the pathogenesis of gestational diabetes mellitus. Determination of HEX activity may have prognostic significance as an early indicator of diabetes mellitus among GDM women in the future.

Lysosomal ß-galactosidase and ß-hexosaminidase activities correlate with clinical stages of dementia associated with Alzheimer's disease and type 2 diabetes mellitus.

Multiple epidemiological studies have shown that individuals affected by type-2 diabetes mellitus (T2DM) carry a 2-to-5-fold higher risk of developing Alzheimer's disease (AD) when compared to non-diabetic subjects. Thus, biochemical parameters that can be easily and routinely assessed for high-confidence evaluation of diabetic conditions leading to AD (AD-T2DM) are regarded as efficient tools aimed at early diagnosis and, in turn, timely AD treatment. In this regard, the activity of lysosomal glycohydrolases may of use, in light of the implication of these enzymes in early events that underlie AD pathology and an overt correlation, in diabetes, between altered metabolic homeostasis, abnormal glycohydrolase secretion in body fluids, and occurrence of diabetic complications. Based on marked up-regulation previously shown in a peripheral, cell-based model of AD, we selected ß-Galactosidase, ß-Hexosaminidase, and α-Mannosidase to discriminate T2DM from AD-T2DM subjects. A screen of 109, 114, and 116 patients with T2DM, AD and AD-T2DM, respectively, was performed by testing enzyme activities in both blood plasma and peripheral blood mononuclear cells. Compared to age-matched, healthy controls (n = 122), ß-Galactosidase and ß-Hexosaminidase activities markedly diverged across the three groups, whereas virtually unchanged values were observed for α-Mannosidase. In particular, plasma ß-Galactosidase and ß-Hexosaminidase levels were higher in patients with AD-T2DM compared to those with T2DM, suggesting different mechanisms leading to enzyme secretion. Statistical analyses based on ROC curves showed that both ß-Galactosidase and ß-Hexosaminidase activities, either intracellular or plasma-secreted, may be used to discriminate AD patients from controls and AD-T2DM from T2DM patients.

IgE, COX-2, and IL-4 are expressed by DEHP through p38 MAPK and suppressed by plant glycoprotein (75 kDa) in ICR mice.

The aim of the present study was to evaluate the inhibitory effect of glycoprotein isolated from Cudrania tricuspidata Bureau (CTB glycoprotein) on di(2-ethylhexyl) phthalate (DEHP)-induced allergic inflammatory response in mice. We evaluated the activity of ß-hexosaminidase, expression of cyclooxygenase (COX)-2, p38 mitogen-activated protein kinase (MAPK), and activator protein (AP)-1, and production of immunoglobulin (Ig)E and interleukin (IL)-4 in DEHP-treated RBL-2H3 cells and ICR mice. Our results revealed that the CTB glycoprotein inhibited the activity of ß-hexosaminidase and production of IgE and IL-4 in serum from DEHP-treated mice. We also found that the CTB glycoprotein reduced arachidonic acid release, COX-2 expression, and AP-1 transcriptional activation through p38 MAPK phosphorylation in DEHP-treated RBL-2H3 cells. The activation of AP-1 was completely blocked by treatment with p38 MAPK inhibitor (SKF86002). The results from these experiments indicate that CTB glycoprotein effectively protects against the allergic inflammation response, mainly through downregulation of MAPK/AP-1 in the mast cell degranulation stage. In conclusion, we suggest that the CTB glycoprotein may be one component of health supplements for the prevention of allergic inflammation.

Inhibition of 2,4-dinitrofluorobenzene-induced atopic dermatitis by topical application of the butanol extract of Cordyceps bassiana in NC/Nga mice.

ETHNOPHARMACOLOGICAL SIGNIFICANCE: The Cordyceps species are insect-borne mushrooms that have been ethnopharmacologically used for skin diseases such as eczema and dermatitis. AIM OF THE STUDY: In this study, we investigated the curative effects of the butanol fraction (CBBF) of Cordyceps bassiana on atopic dermatitis. MATERIALS AND METHODS: Dermatitis was induced by repeated application of 2,4-dinitrofluorobenzene (DNFB) in NC/Nga mice. After a topical application of CBBF on the skin lesions, the dermatitis score, epidermal thickness, mast cell number, and interleukin (IL)-4 and interferon (IFN)-γ, as well as the levels of histamine and immunoglobulin E (IgE) in the serum, were measured. Moreover, effect of CBBF on histamine release was examined using RBL-2H3 under stimulation with 2,4-dinitrophenylated bovine serum albumin (DNP-BSA). RESULTS: CBBF inhibited atopic dermatitis symptoms and signs in the DNFB-treated NC/Nga mice. The suppressive activity of topically applied CBBF may be due to the dose-dependent blockade of a series of immunopathological events, including the release of histamine, the production of IgE, and the secretion of IL-4 and IFN-γ. However, this extract did not directly suppress the degranulation process, assessed by measuring ß-hexosaminidase release. CONCLUSIONS: Our results suggest that CBBF can be applied as an effective herbal remedy to treat atopic dermatitis.

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.