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Dietary supplementation of gamma-linolenic acid (GLA) in the form of a commercial drug neoglandin (containing GLA and vitamin E), in people following alcohol abuse allows bypassing of the ineffective delta-6-desaturase system involved in the transformation of linoleic acid into GLA. Determination of the activity of N-acetyl-β-D-hexosaminidase (HEX) in the serum and urine reflects neoglandin action on the catabolism of glycoconjugates and the functioning of liver and kidneys in people following alcohol abuse. Material and methods: The serum and urine were collected from men with alcohol dependence, treated (n = 31, age 33.16 ± 9.72 years) and not treated (n = 50, age 35.46 ± 11.37 years) with neoglandin. HEX activity were assayed in the supernatants by the colorimetric method, with the p-nitrophenyl derivative of sugar as substrate. Results: Our study on alcoholic men not treated with neoglandin indicates a significantly higher concentration of the serum and urinary HEX activity (nKat/L) on day 1 compared to days 7, 10, 14 and 30 (p < 0.001). For days 14 and 30 (p < 0.01), the urinary HEX activity was expressed in μKat/kgCr. No significant differences were observed in the activity of serum (nKat/L) and urinary (nKat/L and μKat/kgCr) HEX in alcoholics during treatment with neoglandin compared to day 1 of neoglandin treatment. We found significantly different (p < 0.05) concentration of HEX activity (nKat/L) in serum of alcohol-dependent men treated with neoglandin compared to those not taking neoglandin on days 7, 10, 14 and 30 of treatment. The urinary concentration of HEX activity (nKat/L) on days 1, 4, 10 and 30 and HEX activity in μKat/kgCr on days 1, 4 and 7 it was significantly higher (p < 0.05) during the treatment of alcohol-dependence without the use of neoglandin as compared to alcoholics treated with neoglandin. We found a positive correlation between the amount of alcohol consumed and the urinary activity of HEX in the early phase after alcohol withdrawal and a lack of correlation between the HEX activity in serum and urine of alcohol-dependent men not treated with neoglandin. Conclusions: Neoglandin supplementation in alcoholic men significantly slows down the catabolism of glycoconjugates, thus reducing the effects of ethanol poisoning that are harmful to the kidneys. Neoglandin reduces the harmful effects of ethanol poisoning more on the kidneys than on the liver. The activity of HEX in the serum may be used in monitoring the treatment of alcoholism and whether alcohol reuse occurred during the therapy. In the early stages of alcohol withdrawal, urinary HEX activity can be used as a marker of the amount of alcohol consumed during previous alcohol abuse.

Standard indices of renal function, i.e. creatinine, glomerular filtration rate (GFR) and endogenous creatinine clearance, still fail to accurately characterise disease processes in kidneys and are not sufficient for proper therapy selection, efficacy prediction and treatment efficacy monitoring for renal diseases. Therefore, markers that allow for an inexpensive, easy and reproducible kidney function assessment should be sought. Lysosomal exoglycosidases, such as N-acetyl-β-D-hexosaminidase (HEX), its isoenzymes A (HEX A) and B (HEX B), α-fucosidase (FUC), β-galactosidase (GAL), β-glucuronidase (GLU) and α-mannosidase (MAN), excreted in minimal amounts in the urine of healthy children and adolescents, yet showing a significant increase in pathological conditions, may be useful in the diagnosis of kidney disease. Such conclusions were a result of our research on the activity of lysosomal exoglycosidases in the urine of healthy children and adolescents compared to children and adolescents with hypertension, ureteropelvic junction obstruction, and congenital or acquired solitary functioning kidney, as well as based on the findings on the urinary activity of lysosomal exoglycosidases in children and adolescents with various kidney diseases, which were obtained by other authors.

The purpose of the study was to determine the effect of age on lysosomal exoglycosidase activities: α-fucosidase, β-galactosidase, β-glucuronidase and α-mannosidase in healthy children and adolescents.Urine samples were collected from 203 healthy children and adolescents (girls = 99, boys = 104), aged six months to 17.9 years. The activities of α-fucosidase, β-galactosidase, β-glucuronidase and α-mannosidase were determined by colorimetric method and expressed in pKat/μg of creatine (pKat/μg Cr.).Urinary α-fucosidase, β-galactosidase, β-glucuronidase and α-mannosidase activities (pKat/μg Cr.) were the highest in children below 3 years of age in comparison to the remaining age groups. There was a statistically significant negative correlation between urinary α-fucosidase, β-galactosidase, β-glucuronidase and α-mannosidase (pKat/μg Cr.) and age (r = -0.36; r = -0.36; r = -0.35; r = -0.35; at p 0.0001, respectively). In addition, we constructed the reference values for urinary activity of α-fucosidase, β-galactosidase, β-glucuronidase and α-mannosidase (pKat/μg Cr.) in percentiles according to age in 3-year intervals.Our study is the first to show reference values for urinary α-fucosidase, β-galactosidase, β-glucuronidase and α-mannosidase in children and adolescents.

Purpose Adaptation of the colorimetric method for the determination of β- d -galactosidase, β- d -glucuronidase and α- l -fucosidase activities in serums from hemolyzed blood, the material currently being discarded. Materials and Methods The materials included serums from hemolyzed and non-hemolyzed blood, obtained from 26 healthy volunteers. The adaptation of the method involved precipitation of the proteins with trichloroacetic acid after incubating serums with substrates, but before determining the products of enzymatic reactions. Results In serums from hemolyzed and non-hemolyzed blood of the same persons, we found high correlations among the results obtained using hemolyzed blood (with adapted) and non-hemolyzed blood (with non-adapted) methods. Conclusion We are able to determine the β- d -galactosidase, β- d -glucuronidase and α- l -fucosidase activities in serums from hemolyzed blood (with adapted) and non-hemolyzed blood (with non-adapted) methods, with the same accuracy and precision.

OBJECTIVE There is a lack of accurate alcohol-use biomarkers in children/adolescents due to a short drinking duration/rapid normalization of elevated markers. We checked if lysosomal exoglycosidases, elevated earlier in binge-drinking young adults, can be applicable in children/adolescents as markers of harmful alcohol use. METHODS The serum activities (pKat/mL) of α-fucosidase (FUC), β-galactosidase (GAL), β-glucuronidase (GLU), β-hexosaminidase (HEX; its HEX A and HEX B isoenzymes), and α-mannosidase (MAN) were determined in 20 healthy controls (C) and 25 children/adolescents with harmful alcohol use (intoxicated by alcohol at hospital admission -AI1 and on the next day -AI2). RESULTS The serum HEX A and alanine aminotransferase (ALT) activity was significantly higher in the AI1 group than in the control. The activities of FUC, GAL, GLU, HEX B, and MAN were lower in the AI group. We found fair and poor accuracy, respectively, for increased enzymes HEX A and ALT. We found fair accuracy for decreased HEX B (AI1) and MAN (AI1), good accuracy for GLU (AI2), FUC (AI2), GAL (AI1, AI2), MAN (AI2), and excellent for FUC (AI1). Correlations were found: ALT with C-reactive protein (CRP), HEX A with white blood cell (WBC) count, blood alcohol concentration with FUC, MAN and HEX B, and WBC with FUC. CONCLUSIONS Decreased FUC, GLU, GAL, MAN values, and especially FUC (AI1) have the potential to be markers of harmful alcohol use in children/adolescents. The raised activity of HEX A and ALT points to the need for further research to check another inflammatory agent as potential alcohol marker in children and adolescents. Samples need to be collected before intravenous fluid therapy.

The objective of the study was to establish age - dependent values of the urinary lysosomal exoglycosidases activities: N-acetyl-β-D-hexosaminidase (HEX) and its isoenzyme A (HEX A) as well as isoenzyme B (HEX B) in healthy children and adolescents.The study was performed using a random sample of 203 healthy children and adolescents (girls=99, boys=104), aged six months to 17.9 years. The activities of HEX, HEX A and HEX B were determined by a colorimetric method. The activities of the urinary HEX and its isoenzymes were expressed in pKat/μg of creatinine (pKat/μg Cr).Median concentrations of urinary HEX, and its HEX A, HEX B isoenzymes in particular age groups were analyzed using ANOVA. Urinary HEX, HEX A and HEX B activities (pKat/μg Cr) were the highest in children below 3 years, in comparison to remaining age groups. There were statistically significant negative correlations between urinary HEX, HEX A as well as HEX B and age (r=-0.24, p0.001 (HEX); r=-0.20, p0.01 (HEX A); r=-0.26, p0.001 (HEX B), respectively. We constructed the reference values for urinary activity of HEX, HEX A and HEX B (pKat/μg Cr) in centiles according to age, in three-year intervals.Reported data present, for the first time, reference values for urinary activities of HEX and its isoenzymes HEX A and HEX B in children and adolescent.

Reviewing the relevant medical literature on cranial bruits, revealed that the contributions of the Jewish-Polish laryngologist Samuel Meyerson (1851-1939) and his colleague the famous Jewish-Polish neuro- logist Samuel Goldflam (1852-1932), is totally forgotten. This article is a reminder of one of the many historical Polish original contributions to medicine and science.

Aim: Elimination interference of bilirubin on determination serum N-acetyl-β-hexosaminidase (HEX) activity. Material and Methods: We determined HEX activity in sera without and with exogenous bilirubin (4.3 mg/dL) using unmodified and modified method. Modification consists in subtraction absorbance of reaction mixture without substrate, from absorbance of reaction mixture incubated with substrate. Results: HEX activity (nKat/L) amounted: 179.7±74.6 without and 291.4±77.9 with exogenous bilirubin (standard method); 160.5±69.7 without and 164.9±76.8 with exogenous bilirubin (modified method). Correlation coefficient between results of HEX determination in serum without bilirubin and with exogenous bilirubin, obtained with modified method was 0.997. Conclusion: Exogenous bilirubin (4.3 mg/dL) significantly overestimates (p ≤0.001) HEX activity in blood serum determined by the standard method. Proposed modification eliminates interference of bilirubin on determination of HEX activity in blood serum.

OBJECTIVES Investigation of long-term dynamic changes of salivary activity/output of exoglycosidases, deglycosylation processes and their applicability as alcohol markers. METHODS Exoglycosidase (α-fucosidase (FUC), β-galactosidase (GAL), β-glucuronidase (GLU), β-hexosaminidase (HEX, HEX A and HEX B isoenzymes) and α-mannosidase (MAN)) activities were measured in the saliva of healthy social drinking controls (C), alcohol-dependent non-smokers (ANS) and alcohol-dependent smokers (AS) at the 1st, 15th, 30th and 50th day of abstinence after chronic alcohol drinking. RESULTS The activity of exoglycosidases was 2-3-fold (MAN), 2-6 fold (FUC), 8-25-fold (HEX A) and 19-40-fold (GLU) higher in the ANS and AS groups than in controls, and had good/excellent sensitivity, specificity and accuracy. The higher outputs of exoglycosidases were in the AS and ANS groups than in controls at the 1st day (GLU, HEX A) and at the 50th day (GLU, FUC, MAN) of abstinence. We found numerous correlations between alcohol-drinking days with GLU and HEX A, alcohol amounts with HEX A and duration of alcohol dependence with FUC and MAN activity/output. CONCLUSIONS Salivary exoglycosidases/deglycosylation processes were still very high up to 50 days after the end of alcohol consumption. We found markers of chronic alcohol consumption (HEX A), alcohol dependence (FUC and MAN) and chronic alcohol consumption and dependence (GLU).

Pancreatic ductal adenocarcinoma (PDA) accounts for 95% of all pancreatic cancers. About 230,000 PDA cases are diagnosed worldwide each year. PDA has the lowest five-year survival rate as compared to others cancers. PDA in Poland is the fifth leading cause of death after lung, stomach, colon and breast cancer. In our paper we have analysed the newest epidemiological research, some of it controversial, to establish the best practical solution for pancreatic cancer prevention in the healthy population as well as treatment for patients already diagnosed with pancreatic cancer. We found that PDA occurs quite frequently but is usually diagnosed too late, at its advanced stage. Screening for PDA is not very well defined except in subgroups of high-risk individuals with genetic disorders or with chronic pancreatitis. We present convincing, probable, and suggestive risk factors associated with pancreatic cancer, many of which are modifiable and should be introduced and implemented in our society.

Background Determination of lysosomal N-acetyl-β-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 ± 108.76 pkat/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 60 min, pH of reaction mixture was 4.7, and Km was 0.11 mMm. 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.

Purpose Carnitine participates in the metabolism of lipids and cognitive activity. Excessive consumption of alcohol disturbes renal tubular canalicules, that increases urinary excretion of carnitine and its esters. The study evaluates restoration of the urinary free- and total carnitine as well as acylcarnitine excretion after chronic drinking and during the 49-days of controlled abstinence. Materials/methods In 32 patients (6♀; 26♂), 26–60 years old, 2–30 years of alcohol dependence: 75–700 g of pure alcohol (166 ± 94 g) of alcohol daily consumption, 2–360 (35 ± 67) days of intoxication and 1.25 ± 0.8 days of abstinence at admission, we determined urinary free (FC) and total carnitine (TC) as well as acylcarnitine (AC) and acylcarnitine/free carnitine ratio (AC/FC) at admission (T0), after 30 (T30) and 49 (T49) days of the controlled abstinence. Results At T0 excretion of FC, TC and AC as well as AC/FC ratio were significantly higher as compared to the control group. After 30- and 49-days of abstinence, excretion of FC and TC decreased to the level of control group with an exception of the AC and AC/FC ratio at T30 that remained significantly increased. Conclusion 30 days for the FC and TC and 49 days of abstinence for the AC and AC/FC ratio was sufficient to normalize urinary excretion of the carnitines.

Background. Determination of neonate serum’s N-acetyl-β-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.

We investigated the relationship between blood pressure (BP) and the activity of lysosomal exoglycosidases: N-acetyl-β-hexosaminidase (HEX), its isoenzymes A (HEX A) and B (HEX B), α-fucosidase (FUC), β-galactosidase (GAL), β-glucuronidase (GLU) and α-mannosidase (MAN) in pre-hypertensive (high normal blood pressure - HNBP) and normal blood pressure (NBP) children.The study was carried out with urine samples collected from 176 children, aged 6-17.9 years, divided into 2 groups: 42 HNBP and 134 NBP subjects. The children were stratified depending on systolic and diastolic BP (SBP; DBP): HNBP (SBP and/or DBP greater than or equal to the 90th percentile, but less than the 95th percentile) for sex, age, and height; and NBP (SBP and DBP less than the 90th centile). The activities of lysosomal exoglycosidases were determined by the colorimetric method, and expressed in pKat/mL and pKat/μgCr.The activity of urinary HEX A in HNBP group was significantly higher than in NBP (p 0.05). The HNBP group showed significant positive correlation between HEX, HEX A (pKat/mL) and SBP. AUC for HEX A was 0.616, cut-off value -29.351 pKat/mL (sensitivity 51.2%, specificity 71.8%), and 0.589, cut-off value -0.054 pKat/μgCr (sensitivity 31.7%, specificity 86.3%).This is the first report of the relationship between BP and the activity of urinary lysosomal exoglycosidases: HEX, HEX A and HEX B, FUC, GAL, GLU, and MAN in healthy children and adolescents. It seems that HEX A (pKat/mL) can be used as a useful tool in identifying children with HNBP.

AIM Colorectal cancer is characterized by high morbidity and mortality in developed countries. The lack of low-cost, easy-to-use screening diagnostic methods is one of the causes of late diagnosis of colorectal cancer. Beta-glucuronidase (GLU) is a lysosomal exoglycosidase involved in degradation of glycosaminoglycans of the cell membranes and extracellular matrix of normal and cancerous colon tissues. The aim of our research was to evaluate the activity of GLU in the serum of colorectal cancer and estimate its potential value in the diagnosis of colorectal cancer. MATERIAL AND METHODS Blood samples were collected from 21 patients with colorectal adenocarcinoma and 17 healthy subjects. GLU activity was determined by the colorimetric method of Marciniak et al. by measuring the amount of p-nitrophenol released from 4-nitrophenyl-beta-D-glucuronide, at λ = 405 nm. RESULTS We found significantly greater activity of GLU (p

Background We compared effects of binge and chronic alcohol drinking on oral health and salivary immunity proteins. Methods The study involved males: 13 healthy social-drinking (C), 10 alcohol-dependent after chronic alcohol-intoxication (A), and 8 binge-drinkers after a single binge-drinking session (B). We compared periodontal/dental state and salivary immune proteins (lactoferrin –Lf, lysozyme –Lz, oral peroxidase –OPO, immunoglobulin A –IgA) in all groups. Results Group A had worse dental and periodontal states than group C and B. Group B had a lower OPO activity and Lz concentration, and a higher IgA concentration in comparison to group C. Group A had a higher OPO activity than group C. Group B had a lower Lz and a higher LF and IgA outputs than C. Group A had a lower IgA output and a strong tendency of Lf and Lz outputs to be lower than in group C. Positive correlations were found between alcohol amounts and OPO and Lf output in group A, with no such correlations in group B. Only IgA concentration in group B and OPO activity in group A have potential to be markers that help to differentiate binge from chronic alcohol drinking, and OPO activity had better accuracy than IgA. Conclusion Binge alcohol consumption resulted in specific disturbances in salivary innate immunity (Lz), whereas chronic drinking led to disturbances in both adaptive and innate immunity (IgA, Lz and Lf). There is potential applicability of raised salivary IgA concentration and especially OPO activity in binge and chronic drinking detection and differential-diagnosis.

Increasing attention to the importance of saliva testing is not surprising because smoking and alcohol drinking act synergistically on oral tissues, and their metabolite levels, e.g., acetaldehyde, are much higher in saliva than in blood. The activity of salivary alcohol dehydrogenase (ADH) comes from oral microbiota, mucosa, and salivary glands. The purpose of this study was to investigate the involvement of ADH in the oral health pathology of smoking (AS) and non-smoking (ANS) alcohol-dependent males. The results indicated that the AS group had a more significant and longer duration (until the 30th day of alcohol abstinence) decrease in ADH activity and output than the ANS group (until the 15th day of alcohol abstinence) compared to controls (social drinkers; C). The decreased salivary flow (SF) in alcoholics was observed longer in the ANS group (until the 30th day of alcohol abstinence), whereas in the AS group SF normalized at the 15th day, probably due to the irritating effect of tobacco smoke on the oral mucosa. Because saliva was centrifuged to remove cells and debris (including microbial cells), the detected salivary ADH activity was derived from salivary glands and/or oral mucosa. A more profound and longer decrease in ADH activity/output in smoking than non-smoking alcoholics was likely due to the damaged salivary glands and/or oral mucosa, caused by the synergistic effect of alcohol drinking and smoking. The lower values of salivary ADH in smoking than non-smoking alcoholics might also be partly due to the reversed/inhibited ADH reaction by high levels of accumulated acetaldehyde.

Background: Some salivary markers of alcohol abuse/dependence have been proposed so far: aminotransferases, gamma-glutamyltransferase, ethanol, ethyl glucuronide, ethyl sulfate, sialic acid, β-hexosaminidase A, oral peroxidase, methanol, diethylene/ethylene glycol, α-amylase, clusterin, haptoglobin, heavy/light chains of immunoglobulins and transferrin. Aim: To investigate the effect of chronic alcohol drinking and smoking on the activity (pKat/ml) and output (pKat/min) of salivary lysosomal exoglycosidases: α-fucosidase (FUC), α-mannosidase (MAN), β-galactosidase (GAL), and β-glucuronidase (GLU), and their applicability as markers of alcohol dependence. Methods: The activity of FUC, MAN, GAL and GLU was measured colorimetrically in the saliva of healthy social drinkers, alcohol-dependent non-smokers and alcohol-dependent smokers. Results: We observed an increased salivary activity of FUC, GAL, GLU and MAN, as well as an increased output of GAL and GLU, in comparison with controls. The highest increase in the activity/output was found in salivary GLU and MAN (GLU, even 7- to 18-fold), and the least in GAL. We found an excellent sensitivity and specificity and a high accuracy (measured by the area under the ROC curve) for salivary FUC, GLU and MAN activities. The salivary GLU activity positively correlated with the number of days of last alcohol intoxication. Salivary activity of FUC, GAL and MAN, but not GLU, positively correlated with the periodontal parameters such as gingival index and papilla bleeding index. Conclusions: Although we found an excellent sensitivity and specificity as well as a high accuracy for the salivary activity of FUC, GLU and MAN, the GLU activity seems to be mostly applicable as a marker of chronic alcohol drinking (alcohol dependence).

Colorectal cancer (CRC) is one of the most frequent human malignant neoplasms. CRC has an estimated incidence of more than 1,000,000 new cases annually worldwide. Approximately one out of three people who develop CRC dies from the disease. Furthermore, CRC often affects inhabitants of industrialized countries in comparison to less developed countries. Several markers of colon cancer, including CEA, CA-19-9, TPS, TAG-72 and lysosomal hydrolases, have been identified and are now being adopted in routine clinical practice. Increased values of these markers are often the first signal of recurrence or metastases, which is useful in prediction and prognosis of clinical outcome of patients with CRC. Determination of the activity of lysosomal exoglycosidases in body fluids may bring some hope of improving diagnosis of colorectal cancer. However, it has to be remembered that currently the most effective diagnostic method of CRC is endoscopy.

Nasal polyps and hypertrophic lower nasal conchae are common disorders of nasal cavity. The majority of etiopathogenetic theories indicate inflammatory background of polyps and hypertrophic concha. N-acetyl-β-D-hexosaminidase and β-glucuronidase are lysosomal exoglycosidases revealing accelerated activity in inflammatory processes.The aim of the study was to evaluate the catabolism of glycoconjugates in nasal polyps and hypertrophic nasal concha basing on the activity of N-acetyl-β-D-hexosaminidase (HEX) and β-glucuronidase (GLU).Material consisted of nasal polyps taken from 40 patients during polypectomy in patients with chronic rhinosinusitis with nasal polyps (CRSwNP) and hypertrophic lower nasal conchae taken from 20 patients during mucotomy. The activity of HEX, HEX A, HEX B and GLU in supernatant of homogenates of nasal polyps and hypertrophic lower nasal concha tissues has been estimated using colorimetric method.Statistically significant decrease has been observed in concentration of the activity (per 1mg of tissue) of HEX (p0.05), HEX B (p0.001) and specific activity (per 1mg of protein) of HEX B (p0.001) in nasal polyps tissue in comparison to hypertrophic lower nasal conchae tissue.Decrease in the activity and specific activity concentration of the majority of examined lysosomal exoglycosidases (increasing in inflammations) in comparison to hypertrophic lower nasal conchae suggests electrolytes disorders and questions the inflammatory background of nasal polyps.

Background. Severe periodontitis leading to tooth loss is found in 5–15% of most populations worldwide.Aim. The applicability of salivaryβ-hexosaminidase (β-HEX A%, percentage ofβ-HEX A isoenzyme to totalβ-HEX) andβ-HEX B% (β-HEX B/β-HEX) indexes was investigated as a possible marker of periodontitis.Methods. Thirty three alcohol-dependent smokers (AS) and 32 healthy controls (C) were enrolled in the study. The activity ofβ-HEX was measured spectrophotometrically.Results.β-HEX A% was significantly higher andβ-HEX B% was lower in AS than in C group. We found a significant correlation betweenβ-HEX A% and gingival index (GI) and an inverse correlation betweenβ-HEX A% and salivary flow (SF), in all groups. Salivaryβ-HEX A% index in smoking alcoholics at 0.23 had excellent sensitivity (96%) and specificity (91%); the AUC forβ-HEX A% was high (0.937). There were no correlations between amount/duration-time of alcohol drinking/smoking andβ-HEX A% orβ-HEX B%. We found significant correlations between the time period of denture wearing and GI, papilla bleeding index (PBI), and decayed missing filled teeth index (DMFT) and between GI and the amount of smoked cigarettes per day.Conclusion. Bad periodontal state was most likely due to the nicotine dependence. Salivaryβ-HEX A% is a promising excellent marker for the diagnosis of periodontitis.

Introduction Beta-galactosidase (GAL) is a lysosomal exoglycosidase involved in the catabolism of glycoconjugates through the sequential release of beta-linked terminal galactosyl residues. The stimulation of activity of exoglycosidases and other degradative enzymes has been noted in cancers as well as in alcohol and nicotine addiction separately. This is the first study to evaluate the activity of the serum senescence marker GAL in colon cancer patients with a history of alcohol and nicotine dependence, as a potential factor of worse cancer prognosis. Material and methods The material was serum of 18 colon cancer patients and 10 healthy volunteers. Ten colon cancer patients met alcohol and nicotine dependence criteria. The activity of beta-galactosidase (pkat/ml) was determined by the colorimetric method. Comparisons between groups were made using the Kruskal-Wallis analysis and differences evaluated using the Mann-Whitney U test. Spearman's rank correlation coefficient was used to measure the statistical dependence between two variables. Results The activity of serum GAL was significantly higher in colon cancer patients with a history of alcohol and nicotine dependence, in comparison to colon cancer patients without a history of drinking/smoking (p=0.015; 46% increase), and the controls (p=0.0002; 81% increase). The activity of serum GAL in colon cancer patients without a history of alcohol/nicotine dependence was higher than the activity in the controls (p = 0.043; 24% increase). Discussion/conclusion Higher activity of beta-galactosidase may potentially reflect the accelerated growth of the cancer, invasion, metastases, and maturation, when alcohol and nicotine dependence coincide with colon cancer. For a better prognosis of colon cancer, alcohol and nicotine withdrawal seems to be required.

Background: The concentration and specific activity of N-acetyl-β-hexosaminidase (HEX) in palatine tonsils with chronic tonsillitis and tonsillar hypertrophy give insight in tonsillar tissue remodeling and constitute a potential marker for diagnosis and treatment of chronic tonsillitis and tonsillar hypertrophy. Aim: Determining the concentration and specific activity of N-acetyl-β-hexosaminidase in palatine tonsils with hypertrophy and chronic tonsillitis. Methods: HEX activity was analyzed by the method of Marciniak et al. with p-nitrophenyl N-acetyl-β-glucosaminepyranoside as a substrate. Results: The concentration and specific activity of HEX in palatine tonsils in patients with tonsillar hypertrophy and chronic tonsillitis both in childhood and adulthood significantly increase in comparison to healthy individuals. Conclusions: Our data demonstrate the presence of HEX in palatine tonsils and indicate on significant increase of its concentration and specific activity. Based on content and specific HEX activity we suggest that tonsils with hypertrophy and chronic tonsillitis should be treated as identical unit irrespectively of age.

Introduction Nasal polyps are smooth outgrowths assuming a shape of grapes, formed from the nasal mucosa, limiting air flow by projecting into a lumen of a nasal cavity. Up to now the surgical resection is the best method of their treatment, but etiology and pathogenesis of the nasal polyps is not yet fully established. Aim of the study The aim of the study was the assessment of the selected lysosomal exoglycosidases activity in the nasal polyps. In this study the activity of β-galactosidase, α-mannosidase and α-fucosidase was determined in the tissue of the nasal polyps obtained from 40 patients (10 F, 30 M) and control tissues derived from mucosa of lower nasal conchas obtained during mucotomy from 20 patients (10 F, 10 M). Results We observed significant lower values of GAL, FUC and tendency to decrease of MAN and GLU concentration in nasal polyps (P) in comparison to control healthy nasal mucosa (C). In nasal polyp tissue (P) no differences of GAL, MAN and FUC specific activity in comparison to control mucosa (C) were found. Conclusions Our research supports bioelectrical theory of the nasal polyps pathogenesis and directs attention at research on glycoconjugates and glycosidases of the nasal mucosa extracellular matrix.

BACKGROUND Carnitine transports fatty acids from the cytoplasm to the mitochondrial matrix, where the fatty acids are oxidized. Chronic alcohol consumption reduces the concentration of carnitine and interferes with oxidative processes occurring in the cell. AIM The assessment of carnitine concentrations in plasma of chronically intoxicated alcohol dependent persons in a 49-day abstinence period. MATERIAL/METHODS The study included 31 patients (5 women and 27 men) aged from 26 to 60 years (44.6 ± 8.9) and 32 healthy subjects (15 women and 17 men) aged 22-60 years (39.8 ± 9.4). The patients' alcohol dependence ranged from 2 to 30 years (13.6 ± 7.5). Examined subjects consumed 75-700 g of ethanol/day (226.9 ± 151.5). Plasma concentrations of free and total carnitine were measured three times: at the first (T0), 30th (T30) and 49th (T49) day of hospital detoxification. Free (FC) and total (TC) carnitine were determined by the spectrophotometric method. Plasma acylcarnitine (AC) concentration was calculated from the difference between TC and FC; then the AC/FC ratio was calculated. To determine statistically significant differences for related variables, Student's t-test was used. RESULTS At T0, alcoholics had significantly lower concentration of FC and TC (p < 0.05) in plasma, as compared to the control group. In comparison to controls, at T30, plasma TC and FC (p < 0.01) as well as AC (p < 0.001) were reduced. The lowest concentration of TC, FC and AC (p < 0.001)was found at T49. The ratio of AC/FC at T0 had a tendency to be higher in alcoholics than in the control group (p = 0.05), whereas at T49 it was significantly lower in alcoholics as compared to the control subjects (p < 0.05). CONCLUSIONS Chronic alcohol intoxication causes a plasma deficiency of carnitine. Forty-nine days of abstinence showed a significant decrease in the concentration of TC, FC and AC. Further research is necessary to clarify whether a low level of plasma carnitine after chronic alcohol intoxication is caused by the uptake of blood carnitine by tissues such as liver or muscles. In alcoholics the supplementation of carnitine is recommended in the case of a low level of plasma carnitine.

Background A sensitive alcohol marker, β-hexosaminidase (HEX), in the saliva of alcoholics, is investigated for the first time. Methods The activity, specific-activity and output of total HEX and its isoenzymes HEX A and HEX B were measured in the saliva of healthy controls (C), alcohol-dependent non-smokers (ANS), and alcohol-dependent smokers (AS). Results We observed a significantly increased activity/specific-activity and output of HEX A in the ANS and AS groups, due to the inflammatory state of the oral-cavity/salivary-glands. Significantly increased activity of HEX A contributed to an increase in the salivary activity of the total HEX in the ANS group. A significant decrease in the activity/specific-activity of HEX B in AS seemed to be due to HEX B inactivation by cigarette smoke. We noticed a tendency for deteriorated dental state (lower decayed-missing-filled-teeth index – DMFT), worse periodontal state (higher gingival index – GI and papilla-bleeding index – PBI) in AS, and worse periodontal state (higher GI) in ANS, as compared to the controls. We found no differences in the salivary protein concentrations between all groups and decreased salivary flow in both alcoholic groups as compared to the controls. In alcoholics, the area under the curve (AUC) for HEX A activity/specific-activity was significantly greater than for HEX and HEX B. The salivary HEX A activity/specific-activity had good/excellent sensitivity and specificity in smoking and non-smoking alcoholics, whereas salivary HEX and HEX B had poor/fair sensitivity and specificity. Conclusions Salivary HEX A may be helpful in the diagnosis of chronic alcohol intoxication, even in smokers.

Background. The effect of multiple infusions of infliximab (INF), a chimeric anti-tumor necrosis factor alpha antibody, on the concentration of hexosaminidase (HEX) activity in a synovial cell culture derived from human synovial inflamed fluid obtained from patients suffering from rheumatoid arthritis (RA) has been evaluated. Objectives. The aim of this study was to prove INF efficacy in RA. Material and Methods. Inflamed synovial fluid was taken from RA patients (a study group) and patients who had undergone knee trauma within 7 days (a control group). The following solutions of infliximab were used: 40, 60 and 140 µg/mL. Determination of the concentration of HEX activity in cell cultures was performed after 24, 48, 72 and 96 h of infliximab administration. To identify synoviocytes in cell culture immunohistochemical staining with vimentin and pancytokeratin was performed. Results. A predominance of fibroblast-like synovial cells has been observed in the study group. In the control group the concentration of HEX activity without adding infliximab to the cell culture was 283.00 nkat/mL. After 96 h of incubation with infliximab, the concentrations of HEX activity in cultured synoviocytes according to infliximab doses of 40, 60 and 140 µg/mL were respectively: 280.00, 271.50 and 293.50 nkat/mL. In the study group, the concentration of HEX activity without adding infliximab to the cell culture was 542.27 nkat/mL. The final concentrations of HEX activity of cultured fibroblast-like synovial cells measured after 96 h of incubation with infliximab were: 471.72, 498.27 and 556.72 nkat/mL, according to infliximab doses of 40, 60 and 140 µg/mL. In all groups (besides the infliximab concentration of 140 µg/mL after 96 h of incubation), the level of concentration of HEX activity was significantly higher in the study group compared to the control group, irrespective of infliximab concentration and time of infliximab incubation. Conclusions. Infliximab changes the concentration of HEX activity depending on the drug dose and time of administration (Adv Clin Exp Med 2015, 24, 5, 807–813).

The effect of chronic alcohol intoxication and smoking on the output of salivary immunoglobulin A (IgA) was studied in 37 volunteers: 17 male smoking patients after chronic alcohol intoxication (AS) and 20 control non-smoking male social drinkers (CNS). The DMFT index (decayed, missing, or filled teeth), gingival index and papilla bleeding index (PBI) were assessed. Concentration of IgA in saliva was determined by ELISA. Salivary flow (SF) and IgA output were significantly decreased in AS compared to CNS. There were no significant correlations between the amount of alcohol/cigarettes as well as the duration of alcohol intoxication/smoking, and SF or IgA output, nor between IgA level and SF. Gingival index was significantly higher in AS than in CNS, and was inversely correlated with IgA salivary level. The worsened periodontal state in smoking alcohol-dependent persons may result from diminished IgA protection of the oral tissues due to its decreased output.

The aim of the present study was to assess the profiles of the exoglycosidases: N-acetyl-β-hexosoaminidase, β glucuronidase and β galactosidase, α mannosidase and α fucosidase in fibroblast culture undergoing replicative and stress-induced senescence. Half of the cell culture was grown in normal conditions, without the stressor, and the other half of the cell was treated with 0.15 mM tert-butylhydroperoxide. The activities of total N-acetyl-β-hexosoaminidase as well as β glucuronidase in the cell lysate were determined in duplicates using the method of Marciniak et al. The activities of β galactosidase, α mannosidase and α fucosidase in the cell lysate were determined in duplicates using the method of Chatteriee et al. with the modification by Zwierz et al. The activities of the exoglycosidases examined, with the exception of β glucuronidase, showed a significant increase between individual days of the experiment in both non-stressed and stressed fibroblast cell culture. On each day of the experiment, in the cell lysate of stressed fibroblasts, the activities of exoglycosidases were significantly higher compared to the non-stressed cells. There were very strong correlations between SA-β-GAL staining and b galactosidase activity on individual days of the experiment in both non-stressed and stressed fibroblast cell culture. Replicative and stress-induced senescence results in significant changes to the level of lysosomal exoglycosidases, and results in enhanced lysosomal degradative capacity.

Lyme disease (LD) is the most prevalent tick-borne disease in Europe. LD is caused by the spirochete Borrelia burgdorferi. LD is a chronic disease which can attack a number of organs: skin, heart, brain, joints. Chronic, low-grade inflammation involves general production of pro-inflammatory cytokines and inflammatory markers and is a typical feature of aging. So far, the best method of diagnosing LD is a time-consuming and expensive two-stage serological method. The aim of our study was to evaluate the activity of two lysosomal exoglycosidases: α-fucosidase (FUC) and β-galactosidase (GAL) in the serum of patients with Lyme disease, as potential markers of LD. Due to the increasing number of patients with Lyme disease and a number of false results, new ways to diagnose this disease are still being sought. As elevated level of β-galactosidase is a manifestation of residual lysosomal activity in senescent cells, the increase in its activity in serum during chronic Lyme disease might be a marker of a potentially accelerated senescence process. The study was performed on serum taken from cubital veins of 15 patients with Lyme disease and eight healthy subjects (control group). FUC and GAL activity was measured by the method of Chatterjee et al. as modified by Zwierz et al. In the serum of patients with Lyme disease, GAL activity significantly increased (p = 0.029), and the activity of FUC had a tendency to increase (p = 0.153), compared to the control group. A significant increase in GAL activity in the serum of patients with Lyme disease indicates an increased catabolism of glycoconjugates (glycoproteins, glycolipids, proteoglycans) and could be helpful in the diagnosis of Lyme disease, although this requires confirmation in a larger group of patients. As GAL is the most widely used assay for detection of senescent cells, an elevated level of β-galactosidase might be a manifestation of accelerated senescence process in the course of Lyme disease.

Salivary lactoferrin is a glycoprotein involved in the elimination of pathogens and the prevention of massive overgrowth of microorganisms that affect oral and general health. A high concentration of lactoferrin in saliva is often considered to be a marker of damage to the salivary glands, gingivitis, or leakage through inflamed or damaged oral mucosa, infiltrated particularly by neutrophils. We conducted a study to determine the effect of chronic alcohol intoxication on salivary lactoferrin concentration and output. The study included 30 volunteers consisting of ten non-smoking male patients after chronic alcohol intoxication (group A), and 20 control nonsmoking male social drinkers (group C) with no history of alcohol abuse. Resting whole saliva was collected 24 to 48 hours after a chronic alcohol intoxication period. Lactoferrin was assessed by enzyme-linked immunosorbent assay. For all participants, the DMFT index (decayed, missing, or filled teeth), gingival index (GI) and papilla bleeding index (PBI) were assessed. The differences between groups were evaluated using the Mann–Whitney U test. We noticed significantly decreased salivary flow (SF) in alcohol dependent patients after chronic alcohol intoxication (A), compared to the control group (C). Although there was no significant difference in salivary lactoferrin concentration between the alcohol dependent group A and the control group C, we found significantly decreased lactoferrin output in group A compared to group C. We found a significant correlation between the amount of daily alcohol use and a decrease in lactoferrin output. There was a significant increase in GI and a tendency of PBI to increase in group A compared to group C. We demonstrated that chronic alcohol intoxication decreases SF and lactoferrin output. The decreased lactoferrin output in persons chronically intoxicated by alcohol may be the result of lactoferrin exhaustion during drinking (due to its alcohol-related lower biosynthesis or higher catabolism) or to decreased function of neutrophils affected by the ethanol. The poorer periodontal state in alcohol dependent persons compared to controls may be a result of lower salivary flow and decreased protection of the oral cavity by lactoferrin.

The relationship between alcohol consumption and glycoconjugate metabolism is complex and multidimensional. This review summarizes the advances in basic and clinical research on the molecular and cellular events involved in the metabolic effects of alcohol on glycoconjugates (glycoproteins, glycolipids, and proteoglycans). We summarize the action of ethanol, acetaldehyde, reactive oxygen species (ROS), nonoxidative metabolite of alcohol — fatty acid ethyl esters (FAEEs), and the ethanol-water competition mechanism, on glycoconjugate biosynthesis, modification, transport and secretion, as well as on elimination and catabolism processes. As the majority of changes in the cellular metabolism of glycoconjugates are generally ascribed to alterations in synthesis, transport, glycosylation and secretion, the degradation and elimination processes, of which the former occurs also in extracellular matrix, seem to be underappreciated. The pathomechanisms are additionally complicated by the fact that the effect of alcohol intoxication on the glycoconjugate metabolism depends not only on the duration of ethanol exposure, but also demonstrates dose- and regional-sensitivity. Further research is needed to bridge the gap in transdisciplinary research and enhance our understanding of alcohol- and glycoconjugate-related diseases.

There were approximately 93,060 deaths from cancers in Poland in 2008, and about 105,000 are predicted for the year 2025. Early detection of cancer is a major problem throughout the world, which is why many researchers are still looking for specific and sensitive markers of malignant tumors. Our work is a review of recent publications on activity of N-acetyl-β-D-hexosaminidase (HEX) and its isoenzymes A (HEX A) and B (HEX B) as potential markers of malignant tumors. HEX is the most active of the lysosomal exoglycosidases, taking part in degradation of glycoconjugates (glycoproteins, glycolipids, proteoglycans). HEX cleaves N-acetyl-D-glucosamine and N-acetyl-D-galactosamine from non-reducing ends of oligosaccharide chains of glycoproteins, glycolipids and glycosaminoglycans. The activity of HEX, and its isoenzymes A (HEX A) and B (HEX B), was determined by spectrophotometric and isoelectric focusing methods. There was a statistically significant increase in activity of HEX in tumors of the kidney, pancreas, thyroid, colon, ovary, brain, salivary gland, stomach and larynx, which suggests potential applicability of HEX and its isoenzymes in cancer diagnosis.

Up to 30% of all hospital admissions and health-care costs may be attributable to alcohol abuse. Ethanol, its oxidative metabolites, acetaldehyde and ROS (reactive oxygen species), non-oxidative metabolites of alcohol [e.g. FAEEs (fatty acid ethyl esters)] and the ethanol–water competition mechanism are all involved in the deregulation of glycoconjugate (glycoprotein, glycolipid and proteoglycan) metabolic processes including biosynthesis, modification, transport, secretion, elimination and catabolism. An increasing number of new alcohol biomarkers that are the result of alcohol-induced glycoconjugate metabolic errors have appeared in the literature. Glycoconjugate-related alcohol markers are involved in, or are a product of, altered glycoconjugate metabolism, e.g. CDT (carbohydrate-deficient transferrin), SA (sialic acid), plasma SIJ (SA index of apolipoprotein J), CETP (cholesteryl ester transfer protein), β-HEX (β-hexosaminidase), dolichol, EtG (ethyl glucuronide) etc. Laboratory tests based on changes in glycoconjugate metabolism are useful in settings where the co-operativeness of the patient is impaired (e.g. driving while intoxicated) or when a history of alcohol use is not available (e.g. after trauma). In clinical practice, glycoconjugate markers of alcohol use/abuse let us distinguish alcoholic from non-alcoholic tissue damage, having important implications for the treatment and management of diseases.

Pancreatic cancer is the fourth most common cause of death from cancer in the world and the sixth in Europe. Pancreatic cancer is more frequent in males than females. Worldwide, following diagnosis of pancreatic cancer

Objective Determining the activity of lysosomal exoglycosidases in tissue cultures of synoviocytes derived from the knee joints of patients with injured anterior cruciate ligaments (ACL), juvenile idiopathic arthritis (JIA), and rheumatoid arthritis (RA). Methods The following exoglycosidases in cultured synoviocytes were analyzed with p-nitrophenyl derivatives of appropriate sugars as substrates: hexosaminidase (HEX) and its isoenzyme A (HEX-A), β-glucuronidase (GluA), β-galactosidase (GAL), α-mannosidase (MAN), and α-fucosidase (FUC). Results In our cell cultures, fibroblast-like synovial cells (FLS) dominated. In the group of patients with ACL-injuries, and in the groups of patients with JIA and RA, the activity of the investigated exoglycosidases was significantly higher in the intra- rather than in the extracellular compartment. Hexosaminidase was the predominant exoglycosidase. Stimulation of synoviocytes by IL-1β in cell cultures significantly increased the activity of HEX, HEX-A, and GluA in both compartments, as well as of GAL, MAN, and FUC in the intracellular compartment. Stimulation by IL-1β rheumatoidal synoviocytes increased by 128–201% the activity of HEX and HEX A in intracellular compartments and 33–72% in extracellular compartment. Conclusions The profile of lysosomal exoglycosidases in a cell culture of human synoviocytes is similar, but not identical, to those in the knee joint. Hexosaminidase is the dominant glycosidase in cultured unstimulated and IL-1β-stimulated human synoviocytes. The HEX inhibitors may be new drugs for the treatment of inflamed knee joints.

BACKGROUND: The aim of our investigation was to estimate the stability of alpha-tocopheryl O-glycosides in relation to activity of exoglycosidases in selected rat tissues. MATERIAL AND METHODS: Acetylated glycosides were obtained in glucosidation of alpha-tocopherol using the Helferich method. The activity of exoglycosidases was determined by the Zwierz et al. method. Protein concentrations were determined by the biuret method. The concentration of released alpha-tocopherol was determined with the HPLC method. RESULTS: The comparison of the amount of released alpha-tocopherol with the amount of released p-nitrophenol shows that glycoside bound in 2a-5a derivatives of alpha-tocopherol undergoes hydrolysis significantly harder than in appropriate 2b-5bp-nitrophenyl derivatives. CONCLUSION: The results indicate that tocopheryl O-glycosides are more resistant to enzymatic hydrolysis than appropriate p-nitrophenol O-glycosides 2a-5a. Among examined tocopheryl O-glycosides, galactoside 4 is the only compound that caused the significant increase in tocopherol concentration, as compared to its endogenic content.

Our report is the first to show that an acute ingestion (6 h) of a relatively large, yet tolerable dose of alcohol (120-160 g), significantly increases activity of total serum beta-hexosaminidase (total beta-HEX), beta-HEX A and beta-HEX B isoenzymes, as well as salivary total beta-HEX and urinary beta-HEX A, in eight infrequent binge drinkers. An increase in the activity of serum and urinary total HEX is mainly due to its secretory isoenzyme beta-HEX A.

Spontaneously mutated multiple oncogenes and/or tumor suppressor genes in colon epithelial cell and its progeny, may cause proliferation out of control and create benign colon neoplasm (colon polyp). If additional mutations involve genes responsible for cell adhesion and movement, aberrant epithelial cells may become malignant (colon cancer) and invade surrounding and remote tissues, creating secondary tumors called metastases.Incidence of colorectal cancer dramatically increases at 50–65 year of age. In Europe in 2006 colorectal cancer consisted 12.9% of all cancers and caused 207 400 deaths.To laboratory detection and monitoring of colon cancer are used tumor markers. Tumor markers are substances produced by the body in response to cancer, or by cancer tissue itself. Glycoconjugate markers for colon cancer include aberrant: mucins covering the surface of the colon epithelial cells, cadherins, selectins and Ig –like adhesion molecules mediating cell-cell adhesion, integrins and integral membrane proteoglycans responsible for adhesion of colon epithelial cells to extracellular matrix, glycoconjugate components of ECM, as well as lysosomal membrane glycoproteins and exoglycosidases.Detection of colon cancer at early non malignant stage is crucial in its prevention and eradication. As colon cancer is the effect of accumulation many somatic mutations in oncogens, supressors, mismatch repair genes and many genes responsible for posttranslational modifications of proteins, multidirectional approach should be applied for its detection. A glycobiological approach to diagnosis and treatment of colorectal cancer should be directed to detection changes in glycosylation accompanying every step of colon cancer progression, and correlation between changes in glycosylation and tumor progression.