Subject(s)
Appendix , Cecal Diseases , Intussusception , Neoplasms , Appendix/diagnostic imaging , Appendix/surgery , Barium , Cecal Diseases/etiology , Cecal Diseases/pathology , Cecal Diseases/surgery , Humans , Intussusception/diagnostic imaging , Intussusception/etiology , Intussusception/surgeryABSTRACT
Epidermal growth factor receptor inhibitors (EGFRIs) frequently cause cutaneous adverse effects such as papulopustular eruptions. However, the mechanism of the reactions remains unclear. To assess the pathological mechanism of cutaneous adverse reactions caused by EGFRIs, we investigated whether EGFRIs have an influence on the innate immune response of the skin. Levels of human ß-defensins (hBDs), which serve as the first line of defence against infection by pathogenic microorganisms, in the stratum corneum samples of patients treated with EGFR. monoclonal antibodies were measured before and after starting therapy. There were no obvious trends in hBD production in patients without eruptions, whereas a significant decrease in hBD1 and hBD3 production and a nonsignficant decrease in hBD2 production were observed in patients who developed papulopustular eruptions. Our results suggest that a reduction in hBD contributes to the increased incidence of papulopustular eruptions.
Subject(s)
Antibodies, Monoclonal/adverse effects , Drug-Related Side Effects and Adverse Reactions/pathology , ErbB Receptors/antagonists & inhibitors , beta-Defensins/drug effects , Aged , Aged, 80 and over , Anti-Infective Agents/analysis , Anti-Infective Agents/metabolism , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/therapeutic use , Drug Eruptions/etiology , Drug Eruptions/immunology , Drug Eruptions/microbiology , Epidermis/drug effects , Epidermis/metabolism , Epidermis/pathology , ErbB Receptors/immunology , Female , Humans , Immunity, Innate/drug effects , Male , Middle Aged , Staphylococcal Skin Infections/chemically induced , Staphylococcal Skin Infections/epidemiology , beta-Defensins/analysisABSTRACT
BACKGROUND: Neoadjuvant chemoradiotherapy (CRT) is one of the preferred initial treatment strategies for locally advanced rectal cancer. Responses are variable, and most patients still require surgery. The aim of this study was to identify molecular mechanisms determining poor response to CRT. METHODS: Global gene expression and pathway enrichment were assessed in pretreatment biopsies from patients with non-metastatic cT2-4 N0-2 rectal cancer within 7 cm of the anal verge. Downstream Akt activation was assessed in an independent set of pretreatment biopsies and in colorectal cancer cell lines using immunohistochemistry and western blot respectively. The radiosensitizing effects of the Akt inhibitor MK2206 were assessed using clonogenic assays and xenografts in immunodeficient mice. RESULTS: A total of 350 differentially expressed genes were identified, of which 123 were upregulated and 199 downregulated in tumours from poor responders. Mitochondrial oxidative phosphorylation (P < 0·001) and phosphatidylinositol signalling pathways (P < 0·050) were identified as significantly enriched pathways among the set of differentially expressed genes. Deregulation of both pathways is known to result in Akt activation, and high immunoexpression of phosphorylated Akt S473 was observed among patients with a poor histological response (tumour regression grade 0-2) to CRT (75 per cent versus 48 per cent in those with a good or complete response; P = 0·016). Akt activation was also confirmed in the radioresistant cell line SW480, and a 50 per cent improvement in sensitivity to CRT was observed in vitro and in vivo when SW480 cells were exposed to the Akt inhibitor MK2206 in combination with radiation and 5-fluorouracil. CONCLUSION: Akt activation is a key event in the response to CRT. Pharmacological inhibition of Akt activation may enhance the effects of CRT. Surgical relevance Organ preservation is an attractive alternative in rectal cancer management following neoadjuvant chemoradiotherapy (CRT) to avoid the morbidity of radical surgery. Molecular steps associated with tumour response to CRT may provide a useful tool for the identification of patients who are candidates for no immediate surgery. In this study, tumours resistant to CRT were more likely to have activation of specific genetic pathways that result in phosphorylated Akt (pAkt) activation. Pretreatment biopsy tissues with high immunoexpression of pAkt were more likely to exhibit a poor histological response to CRT. In addition, the introduction of a pAkt inhibitor to cancer cell lines in vitro and in vivo led to a significant improvement in sensitivity to CRT. Identification of pAkt-activated tumours may thus allow the identification of poor responders to CRT. In addition, the concomitant use of pAkt inhibitors to increase sensitivity to CRT in patients with rectal cancer may constitute an interesting strategy for increasing the chance of a complete response to treatment and organ preservation.
Subject(s)
Chemoradiotherapy/methods , Neoadjuvant Therapy/methods , Proto-Oncogene Proteins c-akt/metabolism , Rectal Neoplasms/metabolism , Aged , Animals , Blotting, Western , Cell Line, Tumor , Colony-Forming Units Assay , Female , Heterocyclic Compounds, 3-Ring/pharmacology , Humans , Immunohistochemistry , Male , Mice , Mice, Inbred BALB C , Middle Aged , Rectal Neoplasms/therapy , Signal Transduction/drug effects , Treatment OutcomeSubject(s)
Anal Canal/pathology , Anal Canal/surgery , Colonoscopy , Cysts/pathology , Cysts/surgery , Female , Humans , Middle AgedABSTRACT
We studied the effect of suicide gene therapy using an adenovirus vector expressing the cytosine deaminase (CD) gene combined with irradiation therapy (chemo-radio-gene therapy) for human colorectal cancer cells. Since serum CEA levels are elevated in patients with some malignant tumors including colorectal cancer, we applied the CEA promoter to chemo-radio-gene therapy, expecting tumor-specific expression of the CD gene. In in vitro study, we succeeded in selective expression of the target CD gene and growth inhibition in only CEA-producing tumor cells; Further the inhibitory effect was enhanced by combination with radiation therapy in an irradiation dose-dependent manner. In addition, in in vivo study, a significant growth inhibition was observed in chemo-radio-gene therapy in comparison with radiation therapy alone or suicide gene therapy alone. Thus, we suggest that tumor-specific chemo-radio-gene therapy may be a useful strategy for human colorectal cancer.
Subject(s)
Adenocarcinoma/therapy , Antimetabolites, Antineoplastic/pharmacology , Colorectal Neoplasms/therapy , Flucytosine/pharmacology , Genetic Therapy/methods , Nucleoside Deaminases/genetics , Adenocarcinoma/genetics , Adenocarcinoma/metabolism , Adenocarcinoma/radiotherapy , Adenoviridae/genetics , Animals , Antimetabolites, Antineoplastic/pharmacokinetics , Biotransformation , Carcinoembryonic Antigen/biosynthesis , Cell Division/drug effects , Cell Division/genetics , Cell Division/radiation effects , Colorectal Neoplasms/genetics , Colorectal Neoplasms/metabolism , Colorectal Neoplasms/radiotherapy , Combined Modality Therapy , Cytosine Deaminase , Flucytosine/pharmacokinetics , Fluorouracil/pharmacokinetics , Fluorouracil/pharmacology , Genetic Vectors/genetics , HT29 Cells , Humans , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Nucleoside Deaminases/metabolism , Tumor Cells, Cultured , Xenograft Model Antitumor AssaysABSTRACT
We report a very rare early-stage case of mantle cell lymphoma, which arose from the rectum. A 60-year-old man presented with a small elastically hard polypoid lesion in the rectum. The lesion was 1.2 x 1.2 cm in size. As a preoperatively barium enema and endoscopy suggested a benign tumor of the rectum, he underwent local excision of a rectal polypoid mass transanally under spinal anesthesia. However, histological examination revealed a malignant lymphoma, because the lesion was histologically characterized by solid growths of small to medium-sized round cells. Furthermore, immunohistochemical tests revealed B-cell marker positivity and CD5 positivity, but cyclin D1 negativity. Since it was reported that lymphomas with a mantle cell lymphoma morphology and CD5 expression, but without cyclin D1 overexpression, exist in about 10% of mantle cell lymphoma cases, we diagnosed his disease as mantle cell lymphoma. To our knowledge, this is the first reported case of an early-stage mantle cell lymphoma, originating from the rectum.
Subject(s)
Lymphoma, Mantle-Cell/surgery , Rectal Neoplasms/surgery , Biomarkers, Tumor/analysis , Colonoscopy , Diagnosis, Differential , Endosonography , Humans , Lymphoma, Mantle-Cell/diagnosis , Lymphoma, Mantle-Cell/pathology , Male , Middle Aged , Rectal Neoplasms/diagnosis , Rectal Neoplasms/pathology , Rectum/pathology , Rectum/surgerySubject(s)
Adenocarcinoma, Mucinous/surgery , Carcinoma, Intraductal, Noninfiltrating/surgery , Pancreatectomy , Pancreatic Ducts , Pancreatic Neoplasms/surgery , Adenocarcinoma, Mucinous/pathology , Aged , Carcinoma, Intraductal, Noninfiltrating/pathology , Humans , Male , Pancreatic Neoplasms/pathology , SurvivorsABSTRACT
5-Fluorouracil (5-FU) has been used as a chemotherapeutic drug for colorectal cancer. Escherichia coli uracil phosphoribosyltransferase (UPRT), a pyrimidine salvage enzyme, converts 5-FU into 5-fluorouridine monophosphate (5-FUMP) at the initial step of 5-FU activation. We investigated the effects of adenoviral-mediated transfer of the E. coli UPRT gene into human colon cancer cells on 5-FU metabolism and 5-FU chemosensitivity. Three cell lines were used (HT29, KM12 and SW1116). The intracellular levels of 5-fluorodeoxyuridine monophosphate (5-FdUMP) and 5-FU incorporated into RNA after 5-FU treatment in cells infected with adenovirus containing the UPRT gene (AdCA-UPRT) were significantly higher than those of non-infected cells. This was accompanied by marked inhibition of thymidylate synthase (TS) in all cell lines. Furthermore, HT29, KM12 and SW1116 infected with AdCA-UPRT were, respectively, 13.1-, 30.2- and 70.5-fold more sensitive to 5-FU than non-infected cells. Most importantly, treatment with AdCA-UPRT and 5-FU effectively inhibited the growth of HT29-xenografted subcutaneous tumours in nude mice. Therefore, AdCA-UPRT/5-FU treatment had the potential to enhance the actions of 5-FU at both the DNA and RNA levels. Treatment augmented the sensitivity of human colon cancer cells to 5-FU both in vitro and in vivo. We conclude that adenoviral-mediated transfer of the E. coli UPRT gene into colon cancer cells can achieve biochemical modulation of 5-FU and this provides a new approach in the treatment of colorectal cancer.
Subject(s)
Antimetabolites, Antineoplastic/therapeutic use , Colonic Neoplasms/therapy , Escherichia coli/genetics , Fluorouracil/therapeutic use , Genetic Therapy/methods , Pentosyltransferases/genetics , Adenoviridae , Animals , Gene Transfer Techniques , Humans , Mice , Mice, Nude , Tumor Cells, CulturedABSTRACT
The virus-directed enzyme/prodrug system using the Escherichia coli cytosine deaminase (CD) gene and 5-fluorocytosine (5-FC) suffers from a sensitivity limitation in many tumor cells. The E. coil uracil phosphoribosyltransferase (UPRT), which is a pyrimidine salvage enzyme, directly converts 5-fluorouracil (5-FU) to 5-fluorouridine monophosphate at the first step of its activating pathway. To improve the antitumoral effect of the CD/5-FC system, we investigated a combined suicide gene transduction therapy for human colon cancer cells using two separate adenovirus vectors expressing the E. coli CD and E. coli UPRT genes and systemic 5-FC administration (the CD, UPRT/5-FC system). The present study demonstrates that the CD, UPRT/5-FC system generates a co-operative effect of CD and UPRT, resulting in dramatic increases in both RNA- and DNA-directed active forms, including 5-fluorouridine triphosphate incorporated into RNA, 5-fluorodeoxyuridine monophosphate, and the thymidylate synthase inhibition rate, compared with the CD/5-FC system. Furthermore a significant increase in the 5-FC sensitivity of colon cancer cells was demonstrated in the CD, UPRT/5-FC system compared with the CD/5-FC system in vitro and in vivo. These results suggest that the CD, UPRT/5-FC system is a powerful approach in gene therapy for colorectal cancer.
Subject(s)
Adenoviridae/genetics , Colonic Neoplasms/therapy , Escherichia coli/enzymology , Flucytosine/therapeutic use , Genetic Therapy , Nucleoside Deaminases/genetics , Pentosyltransferases/genetics , Adenoviridae/enzymology , Animals , Colonic Neoplasms/genetics , Combined Modality Therapy , Cytosine Deaminase , DNA/analysis , DNA Primers/chemistry , Dose-Response Relationship, Drug , Gene Expression/drug effects , Gene Transfer Techniques , Genetic Vectors , Humans , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , Nucleoside Deaminases/metabolism , Pentosyltransferases/metabolism , RNA/analysis , Tumor Cells, CulturedABSTRACT
Escherichia coli cytosine deaminase (CD), which is a prokaryotic enzyme, converts nontoxic prodrug 5-fluorocytosine (5-FC) into the toxic chemotherapeutic agent 5-fluorouracil (5-FU). To investigate an enzyme/prodrug gene therapy for colorectal cancer, using adenoviral gene transfer of the E. coli CD gene associated with administration of 5-FC, we constructed replication-defective adenovirus vectors expressing the E. coli CD gene or lacZ gene driven by a CAG promoter (composed of a cytomegalovirus immediate early enhancer and a chicken beta-actin promotor). The present study demonstrated that an adenoviral gene transfer system using a CAG promoter induced sufficient gene expression of CD to confer the cytotoxicity of 5-FC to HT29 human colon cancer cells by converting it into 5-FU even at an moi of one. Furthermore, experimental gene therapy using intratumoral injection of the CD-expressing adenovirus with systemical administration of 5'-FC successfully suppressed the growth of established HT29 subcutaneous tumors in nude mice. These results suggest that enzyme/prodrug gene therapy using the adenoviral gene transfer of the E. coli CD gene with concomitant administration of 5-FC may be an effective strategy in the local control of colorectal cancer.
Subject(s)
Colonic Neoplasms/genetics , Colonic Neoplasms/therapy , Flucytosine/therapeutic use , Gene Transfer Techniques , Genetic Therapy , Genetic Vectors , Nucleoside Deaminases/genetics , Nucleoside Deaminases/therapeutic use , Prodrugs/therapeutic use , Adenoviridae , Animals , Cytosine Deaminase , Escherichia coli/genetics , Humans , Mice , Promoter Regions, Genetic , Tumor Cells, CulturedABSTRACT
Many studies have established the usefulness of serum carcinoembryonic antigen (CEA) oriented serial monitoring for predicting recurrence and prognosis; however, few studies have so far investigated serum CEA-negative recurrence. The aim of this study was to elucidate the nature of CEA-negative recurrence regarding tumor angiogenesis. Fifty-seven patients with T3/T4 rectal cancer were divided into the two groups according to the serum CEA status. Angiogenesis was defined as the intratumoral vessel count by immunohistochemical staining using CD31. The CD31 count was significantly higher in the recurrent patients in both groups and the ratio of nodal involvement was significantly higher in the recurrent patients of the CEA-negative group. Local recurrence mainly developed in the CEA-negative group; however, the CD31 count did not predict the sites of recurrence nor the relapse period in the both groups. A multivariate analysis showed a high CD31 count >26) to be a prognostic factor not only for recurrence but also for survival (P = 0.001, 0.043, respectively). These results suggest that a high degree of tumor angiogenesis in sections of T3/T4 rectal cancer may therefore be an important predictor for CEA-negative recurrence.
Subject(s)
Adenocarcinoma/blood supply , Carcinoembryonic Antigen/blood , Neoplasm Recurrence, Local/pathology , Neovascularization, Pathologic/pathology , Rectal Neoplasms/blood supply , Adenocarcinoma/mortality , Adenocarcinoma/pathology , Aged , Female , Humans , Immunohistochemistry , Male , Middle Aged , Multivariate Analysis , Neoplasm Recurrence, Local/mortality , Platelet Endothelial Cell Adhesion Molecule-1 , Rectal Neoplasms/mortality , Rectal Neoplasms/pathology , Rectum/blood supply , Retrospective StudiesABSTRACT
Epidermal growth factor receptor (EGFR) protein overexpression is commonly found in human gastric cancer, and its gene amplification is known to correlate with poor prognosis in gastric cancer patients. With regard to therapy trials targeting EGFR, it has been reported that stable transfection of EGFR antisense or treatment with antibody against EGFR results in growth suppression of human cancer cells that express high levels of EGFR. We have designed an adenovirus-expressing antisense EGFR and have investigated its effect on the growth of gastric cancer in vitro and in vivo. Following infection with EGFR antisense RNA-expressing adenovirus (Ad-EAS), the cell surface EGFR protein levels of infected cancer cells were markedly reduced, and the in vitro growth of Ad-EAS-infected cells was significantly inhibited relative to control-infected cells in all three gastric cancer cell lines (AGS, KKLS, and MKN28) studied here (P < .0002). In a nude mouse subcutaneous tumor system, in vivo tumor growth of MKN28 was significantly inhibited after Ad-EAS treatment, and inhibition on day 48 was 93% by volume compared with that of untreated controls. These results suggest that an adenoviral vector system targeting the down-regulation of EGFR could be a good candidate for the therapy of gastric cancers that overexpress EGFR.
Subject(s)
Adenoviridae/genetics , ErbB Receptors/genetics , Genetic Vectors , RNA, Antisense/pharmacology , Stomach Neoplasms/pathology , Animals , Cell Division/genetics , Humans , Mice , RNA, Antisense/administration & dosage , Tumor Cells, CulturedABSTRACT
The bioavailability of magnesium from Wakame and Hijiki, and the effects of alginic acid on absorption of dietary magnesium were examined in five groups of rats fed either control, Wakame, Hijiki, AW (containing the same amount of alginate as in the Wakame) and AH (containing the same amount of alginate as in the Hijiki) diets, and animals fed a low magnesium diet (LMg) (twentieth amount of magnesium in the original mineral mixtures as the control). Food intake and body weight gain were decreased by adding sodium alginate to the diets. A large amount of calcium accumulated only in the kidneys of the rats fed the LMg diet. Serum magnesium concentration decreased only in the LMg group. The magnesium content in the defatted left femurs did not differ between the control and Wakame fed animals and also among the animals eating Wakame, Hijiki and AW diets. The breaking force of the right femurs did not differ among all the groups except the LMg group. The ratio of apparent magnesium absorption (%) of the control, LMg, Wakame, Hijiki, AW and AH groups was 82.2, 72.7, 66.9, 50.8, 69.3 and 54.2 in the first experimental period, and was 75.3, 52.1, 57.7, 46.9, 62.6 and 60.5 in the second experimental period, respectively. It was clear that the bioavailability of magnesium in the Wakame fed rats was higher than in those eating the Hijiki. Large amounts of sodium alginate lowered magnesium absorption from the diet.
Subject(s)
Alginates/pharmacology , Biological Availability , Magnesium/pharmacokinetics , Seaweed , Alginates/analysis , Animals , Biomechanical Phenomena , Eating/drug effects , Femur/chemistry , Femur/physiology , Glucuronic Acid , Hexuronic Acids , Magnesium/analysis , Male , Minerals/analysis , Rats , Rats, Wistar , Seaweed/chemistry , Weight Gain/drug effectsABSTRACT
To investigate the role of alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) type glutamate receptors in epileptic seizures, we examined the antiepileptogenic and anticonvulsant effects of YM90K [6-(1H-imidazol-1-yl)-7-nitro-2,3-(1H,4H)-quinoxalinedione hydrochloride], a potent and selective new AMPA receptor antagonist, in the rat amygdala-kindling model of epilepsy. Pretreatment with YM90K (7.5-30 mg/kg i.p.) markedly retarded the evolution of kindling. Once kindling was established, administration of YM90K (7.5-30 mg/kg i.p.) significantly and dose-dependently suppressed fully kindled seizures. The maximal effects were observed 15-30 min after injection. When the intensity of electrical stimulation was increased to twice the generalized seizure-triggering threshold, the anticonvulsant effects of YM90K were reversed, suggesting that they were due to elevation of the generalized seizure-triggering threshold. Furthermore, an anticonvulsant dose (15 mg/kg) of YM90K affected neither field potentials nor long-term potentiation in the hippocampus in vivo. These results indicate that AMPA receptors play an important role in the seizure expression mechanism and the development of kindling-induced epileptogenesis, and suggest the possible clinical usefulness of AMPA receptor antagonists as antiepileptic drugs.
Subject(s)
Amygdala/drug effects , Anticonvulsants/pharmacology , Excitatory Amino Acid Antagonists/pharmacology , Hippocampus/drug effects , Kindling, Neurologic/drug effects , Long-Term Potentiation/drug effects , Quinoxalines/pharmacology , Receptors, AMPA/antagonists & inhibitors , Amygdala/physiology , Animals , Dose-Response Relationship, Drug , Hippocampus/physiology , Male , Rats , Rats, Sprague-DawleyABSTRACT
We herein present an extremely rare case of adenocarcinoma of unknown origin arising in the retrorectal space. The imaging and endoscopic findings misled us to preoperatively diagnose a "submucosal tumor." This histological findings were highly suggestive of adenocarcinoma, and its unexpected early and aggressive recurrence required thorough investigation on the true cell type and its origin. Both radiochemotherapy findings and conclusive findings of immunohistochemical staining finally revealed this tumor to be an ordinary adenocarcinoma. It was impossible to identify its origin histologically, because this case was so advanced that it had already replaced the entire entity with adenocarcinoma without leaving any clues as to its former nature. We thus assumed this tumor to be an adenocarcinoma of unknown origin arising in the retrorectal space.
Subject(s)
Adenocarcinoma/pathology , Neoplasms, Unknown Primary/pathology , Pelvic Neoplasms/pathology , Adenocarcinoma/diagnosis , Adenocarcinoma/surgery , Fatal Outcome , Humans , Magnetic Resonance Imaging , Male , Middle Aged , Pelvic Neoplasms/diagnosis , Pelvic Neoplasms/surgery , Tomography, X-Ray ComputedABSTRACT
The present randomized comparative clinical trial was carried out to confirm the efficacy of a newly developed immunosuppressive drug, 15-deoxyspergualin (DSG), and also to find its optimal dose for antirejection pulse therapy. The study included 66 patients with rejection which had occurred within 6 months after renal transplantation. They were treated with DSG at 3, 5 or 7 mg/kg/day for 5 days. The overall percentage of efficacy was 73% and the percentage of efficacy was the highest in the 3-mg/kg group (82%). The incidence of adverse reaction tended to be dose-dependent. From these results, the optimal dose of DSG was judged to be 3-5 mg/kg/day for antirejection pulse therapy, and at doses in this range the percent efficacy could be expected to be 80% or more.
Subject(s)
Graft Rejection/drug effects , Guanidines/therapeutic use , Immunosuppressive Agents/therapeutic use , Kidney Transplantation , Guanidines/administration & dosage , Guanidines/adverse effects , Humans , Immunosuppressive Agents/administration & dosage , Immunosuppressive Agents/adverse effectsSubject(s)
Blood Pressure/physiology , Exercise/physiology , Hypertension/physiopathology , Stress, Psychological/physiopathology , Adult , Aged , Female , Humans , Male , Middle AgedABSTRACT
UNLABELLED: Using a self-imaged diffraction coupled model in a Talbot cavity for vertical cavity surface emitting laser arrays, the effect of self-imaged reflections on the lasing threshold of a finite 2-D array was investigated. Array size and the ratio defined by the element diameter/element spacing were found to affect the effective reflectivity as seen from the laser cavities and, ultimately, the device threshold. A general curve showing the dependence of the 2-D coupling coefficient on the array fill factor and array size has been found. Minimum levels of laser facet reflectivities have been obtained as a function of the array fill factor for practical devices with low threshold current densities. KEYWORDS: 2-D coherent arrays, self-imaging, Talbot effect, laser arrays.
ABSTRACT
Fluorescein isothiocyanate (FITC)-labelled asialotransferrin and pyridyl aminated oligosaccharides were prepared from asialotransferrin and human milk using affinity chromatography and high performance liquid chromatography (HPLC), respectively. These substances were incubated with galactosidase or sialyltransferase and then examined by lectin affinity HPLC. The elution patterns changed according to the period of incubation and amount of enzyme. This analytical method using lectin affinity HPLC with fluorescence labelled glycoprotein or oligosaccharides as the substrates has great value for detecting these enzyme under the same chromatographic conditions. In addition, differences were noted in the activity of beta-galactosidase toward oligosaccharides having the Gal beta(1----3)GlcNAc or Gal beta(1----4)GlcNAc structure at reducing termini.
Subject(s)
Galactosidases/analysis , Sialyltransferases/analysis , beta-Galactosidase/analysis , Chromatography, Affinity/methods , Chromatography, Gel/methods , Chromatography, High Pressure Liquid/methods , Fluorescein-5-isothiocyanate , Fluoresceins , Fluorescent Dyes , Kinetics , Lectins , Magnetic Resonance Spectroscopy , Mass Spectrometry , Plant Lectins , Plants/enzymology , Spectrometry, Fluorescence/methods , ThiocyanatesABSTRACT
Human transferrin was incubated with sialidase and beta-galactosidase and then examined by lectin affinity high-performance liquid chromatography (HPLC). The elution patterns were changed according to the period of incubation and the amount of enzyme. This method of studying lectin affinity HPLC using human transferrin as a substrate makes possible the rapid and important detection of glycosidase activity.