Guidelines for the design and implementation of clinical studies in somatic cell therapy and gene therapy. The German Working Group for Gene Therapy.
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Genetic Therapy↗
Biomedical subjects
S Hase
Publications and source records attributed to S Hase.
Endoscopic assessment of invasion of colorectal tumors with a new high-frequency ultrasound probe.
The present study was carried out to clarify the usefulness of and problems associated with a new ultrasound probe in the assessment of invasion of colorectal tumors. Normal colorectal wall of 23 resected specimens from colorectal cancer cases was examined in vitro with the probe. Fifty-one patients with colorectal carcinoma and 16 patients with rectal carcinoid tumor were presurgically examined using the probe via the biopsy channel of a conventional colonoscope. After endoscopic and ultrasonographic examination, 27 patients underwent endoscopic resection and 40 underwent surgical resection. The ultrasonographic findings were compared with histologic findings in all cases. Carcinomas and carcinoid tumors were visualized as an echo-poor region with the probe. The overall accuracy rate for depth of invasion was 76% (39/51) for colorectal cancer. The accuracy rates for tumors limited to the mucosa and tumors invading the submucosa were 83% and 90%, respectively; these rates were higher than those for tumors invading the muscularis propria (50%) and beyond the muscularis propria (73%). In all 16 cases of carcinoid tumor, the depth of invasion was accurately shown to be limited to the submucosa. This probe is useful to assess invasion of colorectal tumors, especially small and flat lesions limited to the mucosa or submucosa.
Adult↗
The coexistence of esophageal submucosal tumor and carcinoma.
The coexistence of esophageal submucosal tumor and carcinoma has been reported only in very few cases up to now, and the relationship between them is unclear. To elucidate the incidence and the pathogenesis of their coexistence, endoscopy and endoscopic ultrasonography (EUS) were sequentially performed in 95 patients who were suspected of having a submucosal tumor of the esophagus. EUS revealed 83 submucosal tumors and 12 cases of extraluminal compression. Two cases of esophageal carcinoma were observed in the series. In one patient, intraepithelial carcinoma was present on a leiomyoma originating in the muscularis mucosae, and in the other patient, multiple superficial carcinomas were presented.
Aged↗
Endosonographic evaluation of the quality of ulcer healing induced by proton pump inhibitors.
We studied 26 gastric ulcer patients who were treated with a proton pump inhibitor to evaluate the quality of ulcer healing using endoscopic ultrasonography (EUS), and we examined the relationship between ulcer recurrence and contraction demonstrated on ulcer echoes (study I). The effect of lansoprazole versus an H2-receptor antagonist on the contraction of ulcer echoes (study II) was also investigated. In study I, gastric ulcer patients who demonstrated early healing by endoscopy often had shallow Ul-II or Ul-III ulcers and small cross-sectional areas on ulcer echoes. The early contraction on ulcer echo was associated with lower rates of ulcer relapse, and might therefore be indicative of a good quality of healing. In study II, the ulcer contraction rate after 8 weeks of treatment with lansoprazole was 71.8%, significantly higher than that achieved with H2-receptor antagonists. However, the mean cross-sectional area of gastric ulcer after 8 weeks of treatment with lansoprazole was 113.9 mm2. In conclusion, although lansoprazole was suitable for initial therapy in peptic ulcer patients, the quality of ulcer healing based on EUS findings did not appear to be adequate in patients treated for the short periods of time in this study.
2-Pyridinylmethylsulfinylbenzimidazoles↗
[References for planning and implementation of clinical studies in the areas of somatic cell and gene therapy. German Society of Gene Therapy].
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Animals↗
Conversion of egg-white lysozyme to a lectin-like protein with agglutinating activity analogous to wheat germ agglutinin.
Oligomers made of Asp-52-esterified lysozyme or native one showed agglutination for human, mouse, rat and chicken erythrocytes, and also for Hep G2 cells (liver carcinoma-derived cell line) [1] not weaker than wheat germ agglutinin (WGA). Oligomers of Asp-52-esterified lysozyme have about 4-fold stronger agglutinating activity than those of the native one. These results indicate that some enzymes can be converted to lectin-like proteins (neolectin) with binding characteristics similar to their substrate specificity.
Animals↗
A proposed staging system for primary gastric lymphoma.
BACKGROUND: Although primary gastric lymphoma is the most common extranodal lymphoma, no specific staging system exists. METHODS: The authors reviewed 98 cases histologically classified according to the Working Formulation, including low grade B-cell lymphoma of mucosa-associated lymphoid tissue type. Survival rates were calculated by the Kaplan-Meier method according to T, N, and M of the general rules of the International Union Against Cancer TNM system. The definitions of these categories are as follows: T1, tumor invades the lamina propria or submucosa; T2, the muscularis propria; T3, the subserosa; T4, the serosa without invasion of adjacent structures; T5, adjacent structures; N0, no regional lymph node metastasis; N1, perigastric lymph nodes within 3 cm of the edge of the primary tumor; N2, perigastric lymph nodes more than 3 cm from the edge of the primary tumor or in lymph nodes along the left gastric, common hepatic, splenic, or celiac arteries; N3, paraaortic and hepatoduodenal lymph nodes and/or other intraabdominal lymph nodes; N4, beyond N3; M0, no distant metastasis; and M1, positive. RESULTS: The overall 5-year survival rate was 83.5%; it was 100% in T1, 82.4% in T2, 84.2% in T3, 52.9% in T4, and 33.3% in T5; 93.9% in N0, 90.5% in N1, 66.2% in N2, and 44.4% in N3/N4; and 86.0% in M0 and 25.0% in M1. Based on these results, the authors proposed a new staging system as follows: Stage I, T1/N0, N1/M0; Stage II, T1/N2/M0, T2,T3/N0,N1,N2/M0; Stage III, T4,T5/any N/M0, any T/N3, N4/M0; and Stage IV, any T/any N/M1. According to this system, the 5-year survival rate significantly decreased as the stage progressed: 100% in Stage I, 88.9% in Stage II, 52.1% in Stage III, and 25.0% in Stage IV. In Stages I and II, survival rates were not significantly different whether chemotherapy was done or not, whereas in Stage III all patients treated with surgery alone died. CONCLUSIONS: This staging system is useful for assessing prognosis of and deciding a therapeutic plan for primary gastric lymphoma.
Adult↗
High-performance liquid chromatography of pyridylaminated saccharides.
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Aminopyridines↗
Linkage position analysis of pyridylamino-disaccharides by HPLC of fluorogenic Smith degradation products.
Oligosaccharides are often converted to fluorogenic pyridylamino-oligosaccharides (PA-oligosaccharides) to be analyzed sensitively. A method for determining the glycosidic linkage position to the PA-reducing-end residue was developed with PA-disaccharides as model compounds. Periodate oxidation of PA-disaccharides was carried out at 0 degrees C for 15 min or at 4 degrees C for 40 h, and the reaction mixtures were reduced with borohydride. The fluorogenic products obtained at 4 degrees C for 40 h were purified by reversed phase HPLC, and the fraction collected were hydrolyzed with acid. The hydrolysates were analyzed by reversed phase HPLC. PA-glyceraldehyde was formed from 2-substituted PA-disaccharides with PA-hexose, PA-threose (or PA-erythrose) from 3-substituted ones, and PA-glycolaldehyde from 4- or 6-substituted ones. HPLC analysis of the products obtained at 0 degrees C for 15 min revealed a difference between 4- and 6-substituted ones. PA-glyceraldehyde was formed from 6-substituted ones, but not from 4-substituted ones. The linkage position, therefore, can be determined by analyzing fluorogenic product(s). As for PA-disaccharides with PA-N-acetylglucosamine, the linkage position can be simply determined by analysis of 40-h oxidation-reduction mixtures. 2-Acetamido-2-deoxy derivatives of PA-threose, PA-xylose, and PA-glyceraldehyde were formed from 3-, 4-, and 6-substituted ones, respectively. The linkage position analysis was successfully applied to determination of the structures of two Fuc-Man-PAs produced through the transglycosylation action of bovine kidney alpha-L-fucosidase.
Acetylglucosamine↗
A new disaccharide Fuc alpha 1-2Man found in human urine.
Human urine collected from healthy individuals was ultrafiltered and the filtrate was gel-filtered. The fraction including disaccharides was pyridylaminated to convert the reducing sugars to fluorescent pyridylamino (PA)-derivatives. A PA-disaccharide consisting of Fuc and Man was purified by gel filtration, reversed-phase HPLC, and size fractionation HPLC. Structural analysis revealed that the disaccharide was Fuc alpha 1-2Man-PA. The disaccharide is considered to be a metabolite of unknown glycoconjugates.
Carbohydrate Sequence↗
Identification of trisaccharide Xyl alpha 1-->3Xyl alpha 1-->3Glc in human urine.
Oligosaccharides in human urine were converted to pyridylamino (PA)-derivatives. From the PA-oligosaccharides, a saccharide that was chromatographically identical with a synthetic standard, Xyl-alpha 1-->3Xyl alpha 1-->3Glc-PA, was isolated by gel filtration and HPLC. Structure analysis showed that the saccharide was Xyl alpha 1-->3Xyl-alpha 1-->3Glc-PA. It is likely that Xyl alpha 1-->3Xyl alpha 1-->3Glc originated from such glycoconjugates as blood coagulation factors VII and IX, and protein Z.
Carbohydrate Sequence↗
Sequential observation of gastric ulcer healing by endoscopic ultrasonography.
BACKGROUND: The relationship between superficial findings and the inner structure of gastric ulcers is unclear. The present study was undertaken to clarify the relationship between them. METHODS: In the first study, 43 patients with active gastric ulcers were examined by endoscopic ultrasonography (EUS) in each stage of Sakita's endoscopic classification. In the second study, 16 patients with gastric ulcer scars were examined by EUS in both S1 stage and S2 stage. RESULTS: Ulcer area, ulcer length, thickness of the ulcer base, and diameter of the crater differed significantly in each endoscopic stage. Ulcer area, ulcer length, and thickness of the ulcer base differed significantly even between S1 stage and S2 stage. Mean ulcer area in S1 stage was 96.9 mm2; this shows that the ulcer scar in S1 stage is incomplete with regard to tissue contraction. Although ulcer area in S2 stage had shrunk significantly compared with that in S1 stage, it still remained 53.3 mm2. This implies that the risk of local recurrence remains even in S2 stage. CONCLUSIONS: Sequential observation by EUS showed that the healing of the ulcer surface strongly reflected the healing within the ulcers.
Adult↗
Superficial esophageal carcinoma: evaluation by endoscopic ultrasonography.
OBJECTIVES: The aim of this study was to evaluate the usefulness of endoscopic ultrasonography (EUS) in the staging of superficial esophageal carcinoma (SEC). METHODS: We examined the histopathologic findings of 28 patients with SEC which was confirmed in the postoperative histologic evaluation. The EUS results preoperatively estimated were compared with them. RESULTS: There were nine patients with mucosal carcinoma (two intraepithelial carcinomas and seven carcinomas limited to the mucosal layer) and 19 patients with submucosal carcinoma. The mucosal carcinomas revealed no lymph node metastasis or vessel permeation, whereas the submucosal carcinomas revealed lymph node metastasis (71%) and vessel permeation (lymphatic, 58%, vascular, 21%). The accuracy rates of depth of invasion by EUS were mucosa, 67% (6/9); submucosa, 79% (15/19); and total, 75% (21/28). EUS could not detect microinvasion to the submucosa. The overall accuracy rate of EUS in the evaluation of periesophagogastric lymph node metastasis was 72%: sensitivity 58% and specificity 85%. CONCLUSIONS: It has been clinically proved important to differentiate mucosal carcinoma from submucosal carcinoma in the staging of SEC, and EUS is recommended in differentiating between them.
Adult↗
Structures of the N-linked oligosaccharides on porcine plasma vitronectin.
The structures of N-linked oligosaccharides, especially the distribution of sialic acid species, present on porcine plasma vitronectin were elucidated. Oligosaccharides were released from the vitronectin by N-glycosidase F digestion and tagged with 2-aminopyridine, and the pyridylamino-oligosaccharides were fractionated by anion-exchange and reverse-phase HPLC. Nine major pyridyl-amino-oligosaccharides were isolated. The linkages and locations of sialic acids were determined by a novel approach involving desialylation with Salmonella sialidase in combination with acid desialylation. After desialylation, the asialo-forms were analyzed by two-dimensional sugar mapping, component sugar analysis and 400-MHz 1H-NMR spectroscopy. The major oligosaccharides of porcine vitronectin were of the fucosylated biantennary type, with a small amount of the triantennary N-acetyllactosamine type, to which 1-3 mol sialic acids was linked. Sialic acids were linked predominantly through alpha 2-6 linkages, although alpha 2-3 linkages were also present, and fucose was linked to the innermost N-acetylglucosamine through an alpha 1-6 linkage. It was found that every pyridylamino-oligosaccharide population contained N-glycolylneuraminic acid and N-acetylneuraminic acid in a molar ratio of 1:2-9, and that N-glycolylneuraminic acids were located predominantly on the Man alpha 1-6 arm.
Aminopyridines↗
Identification of the characteristic amino-acid sequence for human alpha-amylase encoded by the AMY2B gene.
Human salivary and pancreatic alpha-amylases (HSA and HPA) are the respective gene products of the AMY1 and AMY2A genes. AMY2B is a newly found human alpha-amylase gene. The presence of the AMY2B gene product (HXA) in the urine of healthy humans was examined. A mixture of alpha-amylases that seemed to contain HXA, judging from the substrate specificity, was purified from urine of healthy volunteers by affinity adsorption on starch and then by ion-exchange chromatography. The mixture was reduced and S-alkylated, and the product was digested with trypsin. The digest was separated by reversed-phase HPLC. LVGLLDLALEKDYVR and LVGLLDLALEK, which were found in the digest, are peptides of HXA, but not of HSA and HPA. The detection of these characteristic peptides of HXA demonstrates the presence of HXA in the urine of healthy humans.
Amino Acid Sequence↗
Structures of sugar chains of hen egg yolk riboflavin-binding protein.
The structures of the sugar chains of hen yolk riboflavin-binding protein (RBP) were established. Asparagine-linked sugar chains of yolk-RBP were liberated by hydrazinolysis. Free amino groups of the sugar chains were acetylated and the reducing-end sugar residues were tagged with 2-aminopyridine. Fluorescent pyridylamino (PA-) derivatives of the sugar chains were purified by gel-filtration and reversed-phase HPLC. Seven PA-sugar chains were isolated, and the structure of each was determined by composition analysis, sequential exoglycosidase digestion, methylation analysis, and 500-mHz 1H-NMR spectroscopy. These analyses showed that the main sugar chains had sialylbiantenna and sialyltriantenna structures. PA-sugar chains of plasma-RBP were also isolated, and the structures of the PA-sugar chains of yolk- and plasma-RBPs were compared as to their elution patterns on anion-exchange chromatography and reversed-phase HPLC. The plasma RBP had almost the same sugar chains as the yolk RBP did, indicating that sugar chains are not modified during incorporation into the oocyte.
Animals↗
Structures of N-linked sugar chains expressed mainly in mouse brain.
Sugar chains were liberated from mouse tissues (liver, heart, spleen, kidney, thymus, cerebrum, cerebellum, and brain stem) by hydrazinolysis. After acetylation of the free amino groups, the sugar chains released were pyridylaminated. Pyridylamino (PA-) derivatives of the sugar chains from each tissue were fractionated by anion-exchange high-performance liquid chromatography (HPLC) according to their negative charges. Fractions containing PA-neutral sugar chains thus obtained were separated successively by size-fractionation HPLC and reversed-phase HPLC. Two sugar chains that were more abundant in neural tissues than in other tissues were purified. The structures of the two sugar chains were determined by sugar composition analysis, sequential exoglycosidase digestion, and methylation analysis. The proposed structures are shown below. A structure with n = 1 was found in the cerebrum, cerebellum, and brain stem. A structure with n = 0 was abundant in the cerebrum and brain stem, but less so in the cerebellum. [formula: see text]
Amination↗
Structural analysis of O-linked sugar chains in human blood clotting factor IX.
Type and structural analysis of O-linked sugar chains in human blood clotting factor IX was performed by the pyridylamination method developed for O-linked sugar chains [Kuraya, N. & Hase, S. (1992) J. Biochem. 112, 122-126]. O- and N-linked sugar chains were released with hydrazine, and then N-acetylated, followed by pyridylamination. The type of sugar chain was determined by reducing-end analysis of the pyridylaminated (PA-) sugar chains. Sugar chains with PA-GalNAc at the reducing terminal and that with PA-Fuc [Nishimura, H. et al. (1992) J. Biol. Chem. 267, 17520-17525] were obtained besides known sugar chains with PA-Glc from the Xyl-Glc-Ser type and those with PA-GlcNAc from asparagine-linked sugar chains. The sugar chains with PA-GalNAc were identified as mono- and disialyl Gal beta 1-3GalNAc by two-dimensional HPLC mapping. The structure of the sugar chain with PA-Fuc was Neu5Ac alpha 2-6Gal beta 1-4GlcNAc beta 1-3Fuc, as determined by exoglycosidase digestion, methylation analysis, and Smith degradation.
Carbohydrate Conformation↗