A RAPID AND SENSITIVE METHOD FOR DETECTION OF ARYLSULFATASE ACTIVITIES

Conventional colorimetric methods for determination of arylsulfatase activities are insensitive for microassays. In this study a rapid and sensitive fluorogenic technique was used with issues of the normal levels of arylsulfatase A and B activities in leukocytes. In addition, 2 cases of genetic lysosomal storage diseases were confirmatively diagnosed, one as MLD, the other as MLS, and a third case highly suspected clinically as MLS was finally excluded by this method.

Arylsulfatase,β-galactosidase and lysozyme in gastric cancer cells and its relationship to invasion

IM To study the distribution of arylsulfatase, βgalactosidase and lysozyme in gastric cancer cells, and its relationship to differentiation and invasion of gastric cancer cells.METHODS Histochemical, immunohistochemical and ruthenium red (RR) electrocytochemical technique for three types of hydrolases and proteoglycans in pericancerous matrix in 33 cases of gastric cancer were observed under light and electron microscopy.RESULTS The expression intensities of arylsulfatase, βglactosidase and lysozyme in mucinous cell carcinomas were more intensive than those in welldifferentiated and poorlydifferentiated adenocarcinomas (P<005-001). The fibrous tissues smooth muscle and proteoglycans close to the cancer cells were degraded. They were found in the region far from the cancer cells. Expression of three enzymes mentioned above was low in adenocarcinoma cells, and fibrous tissues and RR granules were present and intact near the welldifferentiated and poorly differentiated adenocarcinoma cells.CONCLUSION Mucinous cell carcinoma may release various hydrolases into extracellular matrix, inducing degradation of pericancerous matrix and facilitating cancer cell invasion and metastasis..

Interaction of arylsulfatases A and B with maspin: A possible explanation for dysregulation of tumor cell metabolism and invasive potential of colorectal cancer

BACKGROUND Although it has been shown that arylsulfatases are lost in colorectal cancer(CRC)cell lines,their exact role in the carcinogenesis and behavior of this cancer was not elucidated.No data about the correlation between serum and immunohistochemical(IHC)level of arylsulfatases(ARSA,ARSB)in patients with CRC were published yet.AIM To evaluate the possible prognostic value of ARSA and/or ARSB in CRC,at circulating and protein levels.METHODS The present study included 45 consecutive patients who were prospectively diagnosed with CRC.For IHC stains(protein expression)ARSA,ARSB and maspin expression were quantified.For these markers,cytoplasmic expression was taken into account.For gene expression study,circulating mRNA was isolated from all patients,before surgery.A group of 45 healthy patients without inflammatory or tumor pathologies was used as control group.Reverse transcription and Taqman Gene Expression Array were used for ARSB gene expression.RESULTS The preoperative circulating RNA level of the ARSB gene was significantly decreased in patients with CRC(RQ<1),compared with the control group(RQ>1).A more significant decrease(RQ<0.5)occurred in ulcero-infiltrative maspinpositive adenocarcinomas,with a higher degree of tumor budding,diagnosed in locally advanced stages(pT3/4).ARSA/maspin immunopositivity indicated a higher risk for lymph node metastasis,while triple positivity for maspin/ARSA/ARSB and ARSB gene expression level<0.5 were indicators of CRC aggressive behavior,independent of lymph node status.CONCLUSION The significant independent negative prognostic factors of CRC are the ulceroinfiltrative aspect,high budding degree,triple positivity for maspin,ARSA and ARSB,and low ARSB gene expression.

Substitution of His260 residue alters the thermostability of Pseudoalteromonas carrageenovora arylsulfatase

This study aimed to improve the thermostability of arylsulfatase from Pseudoalteromonas carrageenovora. A library of P. carrageenovora arylsulfatase mutants was constructed by introducing random mutagenesis using error-prone PCR. After screening, two mutants of H260L and D84A/H260L showed enhanced thermal stability than the wild-type predecessor (WT). Site-directed mutagenesis demonstrated that only amino acid residue at Position 260 plays an important role in the thermostability of P. carrageenovora arylsulfatase. Thermal inactivation analysis showed that the half-life (t1/2) values at 55°C for H260L, H260I, H260Q, H260F and H260R were 40.6, 48.4, 30.9, 29.1 and 34.5 min, respectively, while that of WT was 9.1 min. Structure modeling demonstrated that the additional hydrogen bonds and/or optimization of surface charge-charge interactions could be responsible for the increased thermostability imparted by H260L, H260I, H260Q, H260F and H260R.

Effect of adrenalectomy on rat epididymidis

AIM: To investigate the effect of adrenalectomy (ADX) on the epididymidis of Sprague-Dawley rats. METHODS: The histological, biochemical (cholesterol protein, zinc, copper, alkaline and acid phosphatase aryl sulphatase, lactic dehydrogenase and leucine amino peptidase) and hormonal (FSH, LH and testosterone) changes of caput and cauda epididymis in ADX rats were observed. RESULTS: Organ wet weight, histological studies and morphometric measurements indicated a cellular degeneration in caput and cauda epididymis of ADX rats. Serum testosterone level was significantly lower in ADX than in sham-operated rats, while the serum FSH and LH were below the detection limit of 1 mIU/mL. The enzymatic activity was higher in ADX than in sham-operated rats. Epididymal zinc level increased whereas copper level decreased in ADX rats compared to the sham-operated. CONCLUSION: Adrenalectomy leads to degeneration of caput and cauda epididymidis epithelial cells as a result of decreased supply of testosterone.