GAL Activity Research Articles

Activity of exoglycosidases in blood, urine, cerebrospinal fluid, and vitreous humor in individuals who died from ethyl alcohol poisoning

Lysosomal exoglycosidases, namely α-mannosidase (MAN), α-fucosidase (FUC), β-galactosidase (GAL), and β-glucuronidase (GLU), are of crucial importance in breaking down the oligosaccharide chains of multiple glycoconjugates. Those enzymes liberate monosaccharides from non-reducing ends of oligosaccharide chains. In this study, we have aimed to assess the potential utility of MAN, FUC, GAL, and GLU activities as indicators of ethanol abuse in individuals who died from ethanol intoxication, while also investigating the mechanisms underlying their deaths. The study group comprised 22 fatal ethanol-intoxicated individuals, while the control group included 30 deceased individuals whose body fluids showed no traces of alcohol. We measured the activities (pKat/mL) of MAN, FUC, GAL, and GLU in the supernatants of blood, urine, cerebrospinal fluid as well as vitreous humor. The results indicated significantly lower activities of MAN (p = 0.003), FUC (p = 0.008), GAL (p = 0.014), and GLU (p = 0.004) in the urine of individuals poisoned by ethanol as compared to the control group. Additionally, there was a significantly lower activity of MAN in the vitreous of those affected by ethyl alcohol poisoning (p = 0.016).

Screening of Fabry Disease of patients in renal replacement therapy in a population from Lazio (Italy).

Fabry's disease (FD) is a genetic disorder of lysosomal storage characterized by the intralysosomal accumulation of globotriaosylceramide (Gb3). This genetic mutation causes a total or partial deficit of the α-galactosidase (GAL) enzyme activity. FD has an incidence of 1:40000-60000 born alive. Its prevalence is higher in specific pathological conditions like chronic kidney disease (CKD). The aim of this study was to evaluate the FD prevalence in Italian renal replacement therapy (RRT) patients from Lazio region. 485 patients in RRT (hemodialysis, peritoneal dialysis, and kidney transplantation) were recruited. The screening test was performed on venous blood sample. The latter was analyzed using specific FD diagnostic kit, based on the analysis of dried blood spots on filter paper. We found 3 cases of positivity to FD (1 female and 2 males). In addition, 1 male patient was identified with biochemical alteration indicative of GAL enzyme deficiency with a genetic variant of the GLA gene of unknown clinical significance. The FD prevalence in our population was 0.60% (1 case out 163), it rises to 0.80% (1 case out of 122) if the genetic variant of unknown clinical significance is considered. Comparing the three subpopulations, we observed a statistically significant difference in GAL activity in transplanted patients compared to dialysis patients (p<0.001). Considering the presence of an enzyme replacement therapy able to modify FD clinical history, it is essential to try to implement FD early diagnoses. However, the screening is too expensive to be extended on large scale, due to the low prevalence of the pathology. The screening should be performed on high-risk populations.

Whole-mount Senescence-Associated Beta-Galactosidase (SA-β-GAL) Activity Detection Protocol for Adult Zebrafish.

Senescence-associated beta-galactosidase (SA-β-GAL) is an enzyme that accumulates in the lysosomes of senescent cells, where it hydrolyses β-galactosides. With p16, it represents a well-recognized biomarker used to assess senescence both in vivo and in cell culture. The use of a chromogenic substrate, such as 5-bromo-4-chloro-3-indoyl-β-d-galactopyranoside (X-Gal), allows the detection of SA-β-GAL activity at pH 6.0 by the release of a visible blue product. Senescence occurs during aging and is part of the aging process itself. We have shown that prematurely aged zebrafish accumulate senescent cells detectable by SA-β-GAL staining in different tissues, including testis and gut. Here, we report a detailed protocol to perform an SA-β-GAL assay to detect senescent cell accumulation across the entire adult zebrafish organism ( Danio rerio ). We also identify previously unreported organs that show increased cell senescence in telomerase mutants, including the liver and the spinal cord.

Aging is associated with glial senescence in the brainstem - implications for age-related sympathetic overactivity.

Accumulating evidence suggests that the sympathetic nervous system (SNS) overactivity plays a crucial role in age-related increase in the risk for cardiovascular diseases such as hypertension, myocardial infarction, stroke and heart diseases. Previous studies indicate that neuroinflammation in key brainstem regions that regulate sympathetic outflow plays a pathogenic role in aging-mediated sympathoexcitation. However, the molecular mechanisms underlying this phenomenon are not clear. While senescent cells and their secretory phenotype (SASP) have been implicated in the pathogenesis of several age-related diseases, their role in age-related neuroinflammation in the brainstem and SNS overactivity has not been investigated. To test this, we isolated brainstems from young (2-4 months) and aged (24 months) male C57BL/6J mice and assessed senescence using a combination of RNA-in situ hybridization, PCR analysis, multiplex assay and SA-β gal staining. Our results show significant increases in p16Ink4a expression, increased activity of SA-β gal and increases in SASP levels in the aged brainstem, suggesting age-induced senescence in the brainstem. Further, analysis of senescence markers in glial cells enriched fraction from fresh brainstem samples demonstrated that glial cells are more susceptible to senesce with age in the brainstem. In conclusion, our study suggests that aging induces glial senescence in the brainstem which likely causes inflammation and SNS overactivity.

Senescence associated long non-coding RNA 1 regulates cigarette smoke-induced senescence of type II alveolar epithelial cells through sirtuin-1 signaling

ObjectiveThe primary aim of our study was to explore the mechanisms through which long non-coding RNA (lncRNA)-mediated sirtuin-1 (SIRT1) signaling regulates type II alveolar epithelial cell (AECII) senescence induced by a cigarette smoke-media suspension (CSM).MethodsPharmacological SIRT1 activation was induced using SRT2104 and senescence-associated lncRNA 1 (SAL-RNA1) was overexpressed. The expression of SIRT1, FOXO3a, p53, p21, MMP-9, and TIMP-1 in different groups was detected by qRT-PCR and Western blotting; the activity of SA-β gal was detected by staining; the binding of SIRT1 to FOXO3a and p53 gene transcription promoters was detected by Chip.ResultsWe found that CSM increased AECII senescence, while SAL-RNA1 overexpression and SIRT1 activation significantly decreased levels of AECII senescence induced by CSM. Using chromatin immunoprecipitation, we found that SIRT1 bound differentially to transcriptional complexes on the FOXO3a and p53 promoters.ConclusionOur results suggested that lncRNA-SAL1-mediated SIRT1 signaling reduces senescence of AECIIs induced by CSM. These findings suggest a new therapeutic target to limit the irreversible apoptosis of lung epithelial cells in COPD patients.

The Cardiovascular Phenotype in Fabry Disease: New Findings in the Research Field.

Fabry disease (FD) is a lysosomal storage disorder, depending on defects in alpha-galactosidase A (GAL) activity. At the clinical level, FD shows a high phenotype variability. Among them, cardiovascular dysfunction is often recurrent or, in some cases, is the sole symptom (cardiac variant) representing the leading cause of death in Fabry patients. The existing therapies, besides specific symptomatic treatments, are mainly based on the restoration of GAL activity. Indeed, mutations of the galactosidase alpha gene (GLA) cause a reduction or lack of GAL activity leading to globotriaosylceramide (Gb3) accumulation in several organs. However, several other mechanisms are involved in FD’s development and progression that could become useful targets for therapeutics. This review discusses FD’s cardiovascular phenotype and the last findings on molecular mechanisms that accelerate cardiac cell damage.

P0599THE LYSOSOMAL URINE ENZYMES ACTIVITY IN CHILDREN AFTER ACUTE KIDNEY INJURI

Abstract Background and Aims The urgency of the problem of acute kidney injury (AKI) in children is due to the high risk of chronic kidney disease (CKD) as an outcome. Lysosomal enzymes in urine are considered to be informative markers of renal parenchyma damage. The aim of study was to determine the activity of lysosomal enzymes β-galactosidase (GAL) and N-acetyl-β-D-glucosamynidase (NAG) in urine as markers of CKD progression in children after AKI. Method 41 children were examined after AKI with achieving self-diuresis and clinical improvement. The functional state of the kidneys was evaluated by the glomerular filtration rate (Schwartz GFR), the level of albuminuria. Urine enzyme activity was determined and cross-linked to mmol urinary creatinine. The control (reference) group consisted of 28 children who were conditionally healthy, without kidney disease. Results The NAG activity was 8 times higher level than in the reference group of healthy children (87/46;193 vs. 15/11;18, p&amp;lt;0.001), and GAL was elevated in 3 times either (34/22;45 vs. 10/6;13, p&amp;lt;0.001) in patients with a disease period of up to 2 years after AKI. In patients with follow-up period 2 or more years enzyme activity levels decreased, but remained 4 times higher (NAG: 49/31;109, p &amp;lt;0.001 compared to control; GAL: 35/25;44, p &amp;lt;0.001 compared to control). The high levels of enzyme activity was documented in 73.2% of patients after one year of follow up, but the GFR decrease was found in 22.0%, and albuminuria - only in 34.1%. Conclusion The urine activity of NAG and GAL in children is an informative marker for diagnosing recovery after AKI or CKD progression with renal function decline. This method is non-invasive, inexpensive, and simple to conduct.

Activity of lysosomal urine enzymes in children after acute kidney injury

The importance of the problem of acute kidney injury in children is due to the high risk of developing chronic kidney disease as a consequence. Lysosomal enzymes of β-galactosidase (GAL) and N-acetyl-β-D-hexozoaminidase (NAG) in urine are considered to be informative markers of renal parenchyma damage. The objective of this study - to determine the activity of lysosomal enzymes in urine as markers of progression of interstitial nephritis in children after acute kidney injury.&#x0D; Methods. 41 children were examined after acute kidney injury, achievement of self-diuresis and improvement. Group I included 22 patients with a disease period of up to 2 years after acute kidney injury, group II - 19 patients with a disease period of 2 years or more. The control (reference) group consisted of 28 children who were conditionally healthy, without kidney disease, as well as without acute diseases and severe metabolic disorders and anatomical defects.&#x0D; Results. NAG and GAL activity were found to exceed 8 and 3 times parameters in the reference group of healthy children, respectively, in patients who had acute kidney injury during the year (p &lt;0.001). In patients with a history of 2 years or more, enzyme levels decreased, but remained higher than normal up to 4 times (p &lt;0.001). The highest level of NAG activity was observed in patients of group 1 with the combination of hemolytic-uremic syndrome with hemolytic anemia, and the lowest - in children with toxic kidney.&#x0D; Determination of the activity of lysosomal enzymes NAG and GAL is a more informative marker of torpid course of interstitial nephritis with a progressive decrease in renal function than indicators of glomerular filtration rate. In twelve months after acute kidney damage in 75% of children, the level of NAG and GAL activity remained significantly elevated at normal glomerular filtration rate. A correlation between glomerular filtration rate and NAG activity (r = -0.473) and GAL (r = -0.333) and a direct correlation between NAG and GAL activity (r = 0.845) were observed.&#x0D; Conclusions. The levels of lysosomal enzymes of NAG and GAL were found to be 8 and 3 times higher than normal, respectively, in patients who suffered acute kidney injury during the year, and in patients with a history of 2 years or more, enzyme activity levels decreased, but remained higher than normal 4-fold (p &lt;0.001). In a year after acute kidney damage in 75% of children, the level of NAG and GAL activity remained significantly elevated despite of normal glomerular filtration rate. Therefore, the detection of NAG and β-galactosidase in the urine of children after acute kidney injury are informative markers of renal parenchyma damage, which may serve as objective criteria for progressive decline in renal functional status, and the lack of normalization of their levels is not a sign of progression of interstitial nephritis in corresponding group of children.

Serum Exoglycosidases in Children and Adolescents With Harmful Alcohol Use.

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. 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). 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. 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.

Metabolic process of raffinose family oligosaccharides during cold stress and recovery in cucumber leaves

Raffinose family oligosaccharides (RFOs) accumulate under stress conditions in many plants and have been suggested to act as stress protectants. To elucidate the metabolic process of RFOs under cold stress, levels of RFOs, and related carbohydrates, the expression and activities of main metabolic enzymes and their subcellular compartments were investigated during low-temperature treatment and during the recovery period in cucumber leaves. Cold stress induced the accumulation of stachyose in vacuoles, galactinol in vacuoles and cytosol, and sucrose and raffinose in vacuoles, cytosol, and chloroplasts. After cold stress removal, levels of these sugars decreased gradually in the respective compartments. Among four galactinol synthase genes (CsGS), CsGS1 was not affected by cold stress, while the other three CsGSs were up-regulated by low temperature. RNA levels of acid-α-galactosidase (GAL) 3 and alkaline-α-galactosidase (AGA) 2 and 3, and the activities of GAL and AGA, were up-regulated after cold stress removal. GAL3 protein and GAL activity were exclusively located in vacuoles, whereas AGA2 and AGA 3 proteins were found in cytosol and chloroplasts, respectively. The results indicate that RFOs, which accumulated during cold stress in different subcellular compartments in cucumber leaves, could be catabolized in situ by different galactosidases after stress removal.

Biochemical proxies indicate differences in soil C cycling induced by long-term tillage and residue management in a tropical agroecosystem

A potential benefit of conservation agriculture (CA) is soil organic carbon (SOC) accrual, yet recent studies indicate limited or no impact of CA on total SOC in tropical agroecosystems. We evaluated biochemical indicators of soil C cycling after 9 years (18 seasons) of contrasting tillage with and without maize residue retention in western Kenya. Potential activities of C-cycling enzymes (β-glucosidase, GLU; β-galactosidase, GAL; glucosaminidase, GLM; cellobiohydrolase, CEL), permanganate-oxidizable C (POXC), and soil organic matter (SOM) composition (by infrared spectroscopy) were measured. POXC tended to be greater under reduced tillage and residue retention, but did not significantly differ among treatments (≤ 2% of SOC). Despite no significant differences in SOC concentrations or stocks, activities of all 4 C-cycling enzymes responded strongly to tillage, and to a lesser extent to residue management. Activities of GLU, GAL, and GLM were greatest under the combination of reduced tillage and residue retention relative to other treatments. Reduced tillage produced an enrichment in carboxyl C = O (+6%) and decreased polysaccharide C-O (−3.5%) relative to conventional tillage irrespective of residue management. Though enzyme activities and POXC are typically associated with SOC accrual, changes in soil C cycling at this site have not translated into significant differences in SOC after 9 years. Elevated enzyme activities may have offset potential SOC accumulation under CA. However, the ratio of C-cycling enzyme activities to SOC was higher under reduced tillage and residue retention relative to other treatments, indicating that stoichiometric scaling of SOC and enzyme activities does not explain absence of significant differences in SOC among tillage and residue managements. Potential factors that may explain the low SOC accrual rates in this tropical agroecosystem included the low, albeit realistic, levels of residue retention, nutrient limitations, and high temperatures favoring decomposition.

Monitoring of β-d-Galactosidase Activity as a Surrogate Parameter for Rapid Detection of Sewage Contamination in Urban Recreational Water

Simple, automated methods are required for rapid detection of wastewater contamination in urban recreational water. The activity of the enzyme β-d-galactosidase (GAL) can rapidly (&lt;2 h) be measured by field instruments, or a fully automated instrument, and was evaluated as a potential surrogate parameter for estimating the level of fecal contamination in urban waters. The GAL-activity in rivers, affected by combined sewer overflows, increased significantly during heavy rainfall, and the increase in GAL-activity correlated well with the increase in fecal indicator bacteria. The GAL activity in human feces (n = 14) was high (mean activity 7 × 107 ppb MU/hour) and stable (1 LOG10 variation), while the numbers of Escherichia coli and intestinal enterococci varied by &gt;5 LOG10. Furthermore, the GAL-activity per gram feces from birds, sheep and cattle was 2–3 LOG10 lower than the activity from human feces, indicating that high GAL-activity in water may reflect human fecal pollution more than the total fecal pollution. The rapid method can only be used to quantify high levels of human fecal pollution, corresponding to about 0.1 mg human feces/liter (or 103 E. coli/100 mL), since below this limit GAL-activity from non-fecal environmental sources may interfere.

Effect of Foot and Mouth Disease in Egyptian Cows and Sheep on Characterization of Serum Lysosomal Enzymatic Activities

In this study, clinical findings, virus recognition and the activity of some lysosomal enzymes such as β-N- acetyl glucosaminidase (β- NAG) Acid phosphatase (ACP) and β- galactosidase (β- GAL) in serum of Egyptian dairy cows and sheep naturally infected by foot and mouth diseases were determined. In addition, the lysosomal enzymatic activities in vaccinated animals were also performed. The results revealed that the enzymes activity of ACP was increased by 454% and 806% in vaccinated cows and sheep respectively, while the activity appeared to be less in infected animal either in cow by 182% or 580% in sheep. β -NAG activity exerted a relative lower of the percentage change by 3.3% and 99.0% either in cows or in sheep of vaccinated animals. While in diseased animals the enzyme activity of β- NAG was reduced by 5.6% for cows and increased by 767% for sheep. On the other hand, the enzyme activity of β- GAL appeared to be of moderate activity of enzyme release either in vaccinated or infected animals by 119%, 574% for vaccinated cows and sheep respectively and 54.7%, 1171% for diseased cows and sheep respectively. The obtained results revealed that the enzyme activity of the lysosome was altered according to vaccinated or diseased animals. ACP activity approved to be highly activity in the vaccinated cows than in the diseased ones. The activity of β- NAG in sheep exerted a highly percentage change more than in cows in both of vaccinated and diseased animals. β- GAL activity exerted a highly activity in the diseased animals more than vaccinated animals for sheep. We have concluded that the lysosomal enzymatic activity was highly increased in sheep more than in cows either vaccinated or diseased animals. This study might be open the route for using the lysosomes as adjuvant with FMD vaccine or for increasing the protection against FMD.

Determination of lysosomal exoglycosidases in human saliva.

Currently we observe a growing interest in human saliva as a non-invasive material for diagnosis and monitoring of general and oral diseases. The aim of our study was adaptation of the Marciniak et al. (Marciniak J, Zalewska A, Popko J, Zwierz K, 2006, Clin Chem Lab Med 44: 933-937) method for determination of HEX and GLU activity in synovial fluid, and for determination of: HEX and GLU, as well as MAN, GAL, and FUC activity in human saliva. Under optimal conditions, 10 μl of saliva for HEX, and 30 μl for GLU, MAN, GAL and FUC, were sufficient for determination of human salivary exoglycosidases activity with variation coefficient ranging from 0.89 for GLU to 0.99 for GAL. The adapted method for exoglycosidases activity determination in human saliva is sufficiently sensitive and precise to use in clinical diagnosis.

Salivary Exoglycosidases as Markers of Alcohol Dependence

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. 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. 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. 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. 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).

The activity of serum beta-galactosidase in colon cancer patients with a history of alcohol and nicotine dependence: preliminary data

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. 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. 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). 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.

Lysosomal exoglycosidases in nasal polyps

IntroductionNasal 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 studyThe 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 (10F, 30M) and control tissues derived from mucosa of lower nasal conchas obtained during mucotomy from 20 patients (10F, 10M). ResultsWe 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. ConclusionsOur research supports bioelectrical theory of the nasal polyps pathogenesis and directs attention at research on glycoconjugates and glycosidases of the nasal mucosa extracellular matrix.

Salivary exoglycosidases in gestational diabetes

As exoglycosidases have been described as potential markers of salivary gland pathology, we decided to check the possibility of the use of these enzymes in the detection of salivary gland involvement in gestational diabetes. For this purpose diabetic pregnant women were compared to pregnant and non-pregnant healthy women. The activities of total HEX as well as GLU in the saliva were determined in duplicate according to Marciniak et al. The activities of GAL, FUC, and MAN in the saliva were determined in duplicate according to Zwierz et al. It was found that the specific activities of exoglycosidases in the saliva of diabetic pregnant women significantly increased in comparison to healthy pregnant and non-pregnant women. Increased specific activity of exoglycosidases suggests that gestational diabetes provokes structural/functional alterations in salivary glands and changes in the salivary glycoconjugates metabolism.

Alpha fucosidase and beta galactosidase in serum of a Lyme disease patients as a possible marker of accelerated senescence — a preliminary study

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.

Possible role of α-mannosidase and β-galactosidase in larynx cancer

BackgroundLysosomal exoglycosidases, such as α-mannosidases (MAN) and β-galactosidases (GAL), are found in different glycoside hydrolase sequence-based families. Considerable research has proved plays the role of MAN, which play a key role in the modification and diversification of hybrid N-glycans, processes with strong cellular links to cancer. Therefore the study aim was to investigate the activities of MAN and GAL in larynx cancer compared to controls.Material and methodsLarynx cancer (n = 21) and normal healthy tissue (n = 21) were collected from patients during total laryngectomy. A biopsy of macroscopically healthy tissue in the area of the lower 1/3 of omohyoid muscle was taken for frozen sections in each case and these served as controls. The release of p-nitrophenol from p-nitrophenol derivatives of MAN and GAL was used.ResultsIn all specimens we observed significantly higher activity of investigated enzymes in larynx cancer compared with controls. The mean release of MAN from activated cells was 3.702 ±1.3245 nkat/g wet tissue compared to controls (1.614 ±0.8220 nkat/g wet tissue). The mean release of GAL from the activated cells was 3.383 ±2.1980 nkat/g wet tissue compared to controls (2.137 ±1.3685 nkat/g wet tissue). Differences in observed activity were statistically significant.ConclusionThe present data indicate that MAN and GAL are significantly and consistently elevated in larynx cancer growth. It also means that catabolic reactions involving glycoproteins, glycolipids and proteoglycans may play a role in larynx cancer. Further research should also evaluate the relative importance of these particular exoglycosidases in indicating the progress of the disease in considering the spectrum of identified marker mediators.