Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients: a pilot study.

AIM: Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. This work is focused on glycan structures altered in salivary glycoproteins of alcoholics, indicating the most efficient carriers of such marker glycoepitopes. METHODS: Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. RESULTS: In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. In lectin blotting ten glycoprotein bands were analyzed. The profile of disease-related alterations was found to be complex, but all six lectins studied here were able to detect altered glycan structures. In some glycoproteins the tendency to correct the glycosylation profile was observed after 7 weeks of abstinence. CONCLUSION: Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people were found. Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake.

ICD-10 Classification in the Practice of Emergency Medical Teams: New Insights.

The role of the emergency medical system is to provide assistance to every person in a state in the event of a sudden threat to health and life. Emergency medical teams (EMTs) are an important element of this system, making diagnoses based on the International Classification of Diseases (ICD-10). The study was aimed at analysing the causes of EMT intervention based on groups of diagnoses codified according to the ICD-10. The analysis was based on data from 116,278 EMT interventions in central-eastern Poland in 2017-2019. The research showed that EMT most often made diagnoses based on groups of ICD-10 codes: R00-R99-Symptoms, signs, and abnormal clinical and laboratory findings, not elsewhere classified (39.11%); S00-T98-Injury, poisoning, and certain other consequences of external causes (18.23%); and I00-I99-Diseases of the circulatory system (15.57%). The analysis of the obtained results showed statistically significant differences (p < 0.0001) regarding the area of intervention (urban, rural), sex, age of the patient, and the method of completion of the activities by EMTs in relation to the group of ICD-10 diagnoses for the diagnosis. The conducted study showed the actual reasons for EMT calls. The use of the ICD-10 classification has practical application in EMTs, as it enables the identification of a disease or health problem.

Catabolism of salivary glycoconjugates in acute ethanol intoxication.

BACKGROUND: The aim was to study the effects of a single large dose of ethanol (approximately 2.0 g/kg of body weight, as 40% vodka) on the specific activities of alpha-mannosidase, alpha-fucosidase, beta-glucuronidase, and beta-galactosidase as well as on the total protein concentration in saliva in eight healthy young volunteers. MATERIAL/METHODS: Resting whole saliva samples were collected 12 hours prior to and 36 and 108 hours after alcohol consumption. Exoglycosidase activities were assayed in the supernatants by the colorimetric method. Protein content was determined by the Lowry method. RESULTS: Thirty-six hours after alcohol consumption the specific activities of alpha-fucosidase and beta-glucuronidase were significantly higher than before drinking. The specific activity of beta-galactosidase showed a greater tendency to increase than alpha-mannosidase after the drinking session. The total protein concentration was significantly lower after alcohol consumption than at baseline, even at 108 hr. Significant inverse correlations between total protein content and the specific activities of the exoglycosidases in saliva were found after the drinking session. CONCLUSIONS: Acute ingestion of a large dose of ethanol increased the activity of salivary exoglycosidases, which might be followed by subsequent degradation of proteins in saliva. The observed changes might contribute to salivary defense system malfunction as well as to oral malodor production.

[Fructose-1,6-bisphosphatase--marker of damage to proximal renal tubules]. / Fruktozo-1,6-bisfosfataza--marker uszkodzenia cewek nerkowych blizszych.

Pathological processes disturbing function of renal proximal tubules, increase activity of fructose-1,6-bisphosphatase (FBP-1) in urine. FBP-1 is cytosolic enzyme which occured mainly in cells of proximal renal tubules, and to small extent in cells of pars recta. After damage to the cell membrane FBP-1 is more rapidly excreted to the urine, than enzymes residing in other cell organelles. Fructose-1,6-bisphosphatase was isolated from rabbit muscle in 1943 by Gomori, and from spinach in 1958 by Racker i Schröder. Highest activity of FBP-1 was found in liver and kidneys, lesser in ileum, leucocytes, muscles and brain. Fructose-1,6-bisphosphatase is one of four key enzymes of gluconeogenesis performing synthesis of glucose from non sugar substrates. FBP-1 catalyses hydrolysis of fructose-1,6-bisphosphate in cytoplasm of the cell. There are many reports on properties and significance of FBP-1 in plant and animal tissues, but only few reports on activity of this enzyme in urine. Reason for little interest in determination of FBP-1 activity in urine, is relative instability of this enzyme in urine.

[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.

Analysis of the relationship between cancer procoagulant activity and PCNA and Ki-67 expression in cases of common and cellular uterine leiomyomas.

PURPOSE: Histological subtypes of uterine leiomyomas may substantially differ in their cellular biology, including the intensity of synthesis of cancer markers and expression of cell proliferation markers. The present investigation aimed to determine the activity of cancer procoagulant (CP) in subtypes of leiomyomas, including cellular leiomyomas, and to verify whether these activities correlate with immunoexpression of cell proliferation markers: the proliferating cell nuclear antigen (PCNA) and Ki-67. MATERIALS AND METHODS: Preoperative peripheral venous blood and postoperative tissue material were obtained from 24 women operated on in a tertiary referral academic department. The activity of CP in serum was measured with the use of a coagulative method according to Gordon and Benson, and in tissue homogenates with the use of a spectrophotometric method according to Colucci et al. The control serum values were obtained from 20 healthy women without any gynecological disease, and the control solid tissue values from histologically confirmed postoperative normal reproductive tissues obtained from six patients. PCNA and Ki-67 expression were determined immunohistochemically using monoclonal antibodies. RESULTS: Both the tissue and serum activity for CP was considerably higher for common leiomyomas and cellular leiomyomas than for control tissues, but did not differ significantly between the leiomyoma subtypes. Intratumor CP activity significantly correlated with PCNA expression but not with Ki-67 expression. CONCLUSIONS: Cellular leiomyomas do not differ substantially in the serum and intratumor CP activity from common leiomyomas. There is a relationship of intratumor CP activity with PCNA expression, a finding which requires further investigation.

[Concentration of thyroid stimulating hormone and activity of N-acetyl-beta-D-hexosaminidase and its isoenzymes, in serum of patients with thyroid cancer]. / Stezenie hormonu tyreotropowego oraz aktywnosc N-acetylo-beta-D-heksozoaminidazy i jej izoenzymów A i B w surowicy chorych na raka tarczycy.

UNLABELLED: Thyroid cancer consists 1% of all malignant neoplasms. It is not known interrelationship between concentration of TSH in blood serum and condition of thyroid cancer. Thyroid cancer is difficult for diagnosis and differentiation. Therefore it is necessary to search for biochemical markers helpful in diagnostics of thyroid cancer. Significant increase in activity of N-acetyl-beta-D-hexosaminidase and its isoenzymes A and B in serum of patients with neoplasms of kidney and pancreas suggest approporiateness of evaluation of HEX and its isoenzymes in diagnostics of thyroid cancer. THE AIM: of the study--evaluation of TSH concentration and activity of HEX and its isoenzymes A and B, in serum of patients with thyroid cancer. MATERIALS AND METHODS: Blood was taken from 7 patients with thyroid cancer (6 men and 1 woman). Control consisted of 7 healthy men. In blood serum concentration of TSH was determined with immunoenzymatic method on analyzer Axsym of Abbott and expressed in microU/mL. The activity of HEX and its isoenzymes A and B was determined by method of Chatterjee et al., as modified by Zwierz et.al. Determination of HEX was performed on microplate reader ELX800 BIO-TEK. Activity of HEX, HEX A and B was expressed in pKat/mL, and specific activity in pKat/mg protein). Protein was determined by biuret method and results were expressed in mg/mL. RESULTS: Concentration of HEX A activity in serum of thyroid cancer patients is significantly higherin comparison to healthymen (p = 0.0191). Also specific activity of HEX A in serum of thyroid cancer patients is significantly higher in comparison to healthy men (p = 0.0393). CONCLUSIONS: 1. Determination of TSH concentration in serum of thyroid cancer before the operation may confirm euthyreosis. 2. Determination of HEX A activity in serum may be helpful in diagnostics of thyroid cancer.

[Usefulness of detecting cancer procoagulant activity and thyrotropic hormone concentration in the differentiation of tumor-like changes in the thyroid]. / Zastosowanie badania aktywnosci nowotworowego prokoagulanta oraz stezenia hormonu tyreotropowego w róznicowaniu zmian guzowatych tarczycy.

Epidemiological studies have shown the increased incidence of malignant cancer of the thyroid gland observed in the last decade. This increase is connected with the elevated number of benign tumor-like/tuberous changes in the thyroid gland. Since it is difficult to differentiate diagnostically this pathology, it would be justified to search for biochemical markers which can help to confirm this change. The aim of our study was to evaluate the usefulness of detecting cancer procoagulant activity (CP) and thyrotropic hormone concentration (TSH) in the differentiation of tumor-like changes in the thyroid gland. The study included 15 patients (12 women and 3 men) with adenoma glandulae thyreoideae or nodular changes in the character of struma nodosa hyperplastica and 12 patients (11 women and 1 man) with carcinoma glandulae thyreoideae. A control group consisted of 12 healthy people (5 women and 7 men). CP activity was determined in the serum by the coagulation method according to Gordon and Benson and it was expressed as the coagulation time in seconds (s). TSH concentration was measured by the immunoenzymatic method (MEIA) using an analyzer of Axsym (Abbott) and was expressed in microU/ml. The results of our study indicate that the determination of CP activity can be used in the differential diagnosis of tumor-like changes of the thyroid gland. The concentration ofTSH was within the normal values, despite statistically different mean values between particular groups that results from the fact that patients qualified to surgery were in the state of euthyreosis.

Lysosomal exoglycosidases and cathepsin D in colon adenocarcinoma.

INTRODUCTION: Changes in the structure of membrane glycoconjugates and activity of glycosidases and proteases are important in tumor formation. OBJECTIVES: The aim of the study was to compare the specific activity of lysosomal exoglycosidases: N-acetyl-ß-D-hexosaminidase (HEX), its isoenzymes A (HEX A) and B (HEX B), ß-D-galactosidase (GAL), α-fucosidase (FUC), and α-mannosidase (MAN) with the activity of cathepsin D (CD) in serum, urine, and carcinoma tissue of patients with colon adenocarcinoma. PATIENTS AND METHODS: The specific activity of HEX, HEX A, HEX B, GAL, FUC, MAN, and CD was assayed in serum, urine, and carcinoma tissue of 12 patients with colon adenocarcinoma. RESULTS: Lysosomal exoglycosidases and CD have similar specific activity in colon adenocarcinoma tissue and urine, which is higher than their activity in serum (with the exception of the highest specific activity of CD in urine). A positive correlation was observed between the specific activity of CD and that of HEX, HEX A, FUC, and MAN in the carcinoma tissue and urine as well as between CD and GAL in the urine of patients with colon adenocarcinoma. Negative correlations were observed between protein levels and the specific activity of HEX, HEX A, FUC, MAN, and CD in the carcinoma tissue and urine, and between protein levels and GAL in urine. CONCLUSIONS: Increased degradation and remodeling of glycoconjugates in the colon adenocarcinoma tissue is reflected by increased specific activity of exoglycosidases and CD. The results suggest a strong effect of exoglycosidase action on tissue degradation and a potential role of exoglycosidases in the initiation of proteolysis.

The activity of lysosomal exoglycosidases in serum of alcohol-dependent men supplemented with borage oil enriched with vitamin E.

The aim of this study was to determine the activity of the lysosomal exoglycosidases: alpha-mannosidase (MAN), alpha-fucosidase (FUC), and beta-glucuronidase (GLUCUR) in serum of alcohol-dependent men supplemented and not supplemented with borage oil enriched with vitamin E. Serum was collected from eight social drinkers and 16 alcohol-dependent men after a drinking period. The activity of exoglycosidases and the concentration of protein in serum were determined. The increase in specific activity of MAN and GLUCUR was significant in serum of alcohol-dependent men both not supplemented and supplemented with borage oil enriched with vitamin E, in comparison with the specific activity in serum of social drinkers. In serum of alcohol-dependent men treated with borage oil enriched with vitamin E, specific activity of MAN and GLUCUR fluctuated in comparison with alcohol-dependent men not supplemented. Specific activity of FUC in serum of alcohol-dependent men both not supplemented and supplemented with borage oil enriched with vitamin E showed a tendency to increase, in comparison with social drinkers. Specific activity of FUC had a tendency to decrease in serum of alcohol-dependent men supplemented with borage oil enriched with vitamin E, in comparison with alcohol-dependent men not supplemented. Thus, supplementation of alcohol-dependent men after a long-lasting drinking period with borage oil and vitamin E did not change the rate of catabolism of the oligosaccharide chains of glycoconjugates, as evaluated by serum activity of exoglycosidases.