Sialyi Lex (FH6) and sialyl Lea (CA19—9) antigens, members of a family of high molecular weight glycoproteins, are ligands for E-selectin and may play important roles in tumor metastasis. However, expression patterns of sialyl Lex and sialyl Lea have not been established in human extrahepatic bile duct cancer. In this study, we examined sialyl Lex and Lea expression in human extrahepatic bile duct adenocarcinoma and their clinicopathological significance. Sialyl Lex immunoreactivity was detected not only in cancer cells (cytoplasmic type; 57.7%, 30/52), but also in cancer stroma (stromal type; 19.2%, 10/52). Sialyl Lea immunoreactivity was found in cancer cells (cytoplasmic type; 84.6%, 44/52) and in cancer stroma (stromal type; 63.5%, 33/52). Stromal sialyl Lea expression was more frequent than stromal sialyl Lex expression (p<0.001). According to TNM classification, cytoplasmic sialyl Lex and sialyl Lea expression was detected in 62.5% (30/48) and 85.4% (41/48) of the T2-3 cancers (T2-3 vs T1, sialyl Lex, p = 0.015; and sialyl Lea, p = 0.310). Cytoplasmic sialyl Lea positive cancers frequently showed lymph node metastasis (p = 0.029). These obseivations suggested that sialyl Lea expression plays more important roles than sialyl Lex in metastasis of human extrahepatic bile duct adenocarcinoma.
We examined the influence of intravenous injection of glycine on the spinobulbospinal and spinal micturition reflexes in rats. Female rats were divided into an intact group and a chronic spinal cord injury (SCI) group with transection of the lower thoracic cord, and studied at 4 weeks after SCI. Under urethane anesthesia, isovolumetric cystometly was performed in each group before and after intravenous injection of glycine (0.01-100 mg/kg). Intravenous injection of glycine (0.1-100 mg/kg) transiently abolished bladder contractions after a delay of several minutes in both groups. In intact rats, intravenous injection of glycine prolonged the interval between bladder contractions, but did not change the amplitude of the contractions. On the other hand, intravenous glycine both prolonged the interval and decreased the amplitude of bladder contractions in SCI rats. However, intrathecal injection of low dose of strychnine (a selective glycine receptor antagonist; 0.001 μg) and following intravenous injection of glycine (0.1 mg/kg) did not affect the bladder contractions in intact rats. These results suggest that systemically administered glycine inhibits the afferent limb of the spinobulbospinal micturition reflex, and inhibits both the afferent and efferent limbs of the spinal micturition reflex at the lumbosacral cord level.
IgG induces both passive and active anaphylaxis but the role of IgG glycosylation in inducing anaphylaxis remains unclear. We sensitized animals to enzymically deglycosylated and native anti-OVA IgG1. Native IgG1 induced passive cutaneous anaphylaxis in Sprague-Dawley rats, showing dye leakage on the rat back, and passive systemic anaphylaxis in ddY mice, showing marked blue coloring of foot pads, significant decreases in rectal temperature and killing all ddY mice within 50 minutes of antigen challenge, while deglycosylated IgG1 completely lost its allergic activity. The abrogation of deglycosylated IgG1 to induce anaphylaxis was not due to the loss of its antigen binding ability because ELISA assay showed that native and deglyeosylated IgG1 had the same ability to bind antigen. Neutralization of lethality in OVA-sensitized mice was found when deglycosylated IgG was intravenously injected 24 hours before challenge with OVA. These results suggest that IgG1-induced anaphylaxis requires IgG1 glycosylation. Deglycosylated IgG may have therapeutic potential for antigen-specific anaphylaxis.
CD4+ helper T (Th) cells are subdivided into Th1 and Th2 cells in terms of their differential cytokine production pattern. These cells are key immunoregulateiy cells in host defense mechanisms. Therefore, identification of differential gene expression in Th1 and Th2 cells is crucial for immunomodulation. Here we describe identification of Th1- or Th2-specific genes by DNA microarray analysis. We prepared mRNA from anti-CD3 mAb stimulated or unstimulated Th1 and Th2 cells, then gene expression profiles were analyzed by using a microarray spotted with 8,700 EST clones. We detected preferential expression of 57 genes in Th1 cells and 37 genes in Th2 cells. By semi-quantitative RT-PCR analysis, we identified 11 genes specifically expressed in Th1 cells, and 7 genes in Th2 cells. These genes may contribute to the differentiation and function of helper T cells and thereby regulation of immune response. Further analysis would help to develop a method to modulate immune response.
The purpose of this study was to examine the ventilatory responses during a six-minute walk test (6MWT), an incremental shuttle walking test (ISWT), and a cycle ergometer test (CET) in patients with chronic obstructive pulmonary disease. Twelve subjects (11 male, age 72.2 years, with a forced expiratory volume in one second of 53.6% of the predicted normal) took part in the study. Throughout the three exercise tests, oxygen uptake (V'Ｏ2), carbon dioxide production (V'CO2), minute ventilation (V'E), and heart rate (HR) were measured using a portable breath-by-breath system. Oxygen saturation (SpO2) and breathlessness were recorded at the end of each minute during the tests. There were no significant differences among the three exercise tests in terms of the peak values for V'O2, V‘CO2, V'E, HR, and breathlessness. SpO2 was significantly lower in both walking tests (6MWT: 89.8 ± 3.3%, ISWT: 88.9 ± 3.8%) compared to values in the CET (93.3 ± 2.3%) (p < 0.05). V'O2 during the 6MWT increased sharply for 2 minutes, and then showed a steady-state profile. During the CET and the ISWT, however, V'O2 increased with respect to workload. In conclusion, the three tests clearly measure different aspects of exercise tolerance and should be considered complementary.
We have proved before that μ-calpain can be a promising autogenous agent to replace chymopapain in chemonucleolysis treatment. Even in the case of accidental intrathecal injection, the undesirable neural damage of calpain can be neutralized by normal cerebrospinal fluid (CSF). The aim of this study was to characterize this unknown calpain inhibitor-like component in the CSF of normal rabbits. A heat-stable low molecular weight fraction in the CSF was identified. It was not extracted by chloroform solution, but labile to protease digestion. It inhibited the calpain proteolysis of microtubule-associating protein-2 (MAP-2) and membrane structure proteins of platelets and PC12 cells. It can block the autolytic activation of native human erytluocyte μ-calpain, where the conversion of the large subunit of μ-calpain from the 80-kDa form to the 76-kDa form was obstructed, with accumulation of the intermediate 78-kDa form. A possible calpain inhibition mechanism was discussed. After purification by paper chromatography, gel filtration and reverse phase chromatography, the composition of the calpain inhibitor was investigated. An oligopeptide with molecular weight about 800 Da was identified as a novel calpain inhibitor by partial amino acid sequence.
The purpose of this study was to clarify the effects of peroxide on teeth after the tooth-whitenerapplication. An artificial root canal was prepared using the crown of human incisors filled with phosphate buffer solution. Tooth-whitener containing 30% hydrogen peroxide or 35% carbamide peroxide was applied according to the instructions with or without acid-etching pretreatment. Luminol-chemiluminescence was used to measure the quantity of peroxide that penetrated into the buffer within the root canal. The penetrated peroxide quantity ranged from 7 μM to 100.5 μM. The penetration quantity for the hydrogen peroxide-applied group was about 1.6-fold greater than that for the carbamide-applied group. The penetration quantity for the acid-etching pretreatment group exceeded about 4-fold greater than that of the non-pretreatment group. The whitener containing hydrogen peroxide consists of liquid and powder. Qualitative analysis of powder component of the whitener with EPMA showed that it contained Fe and Cu ions. This finding suggests that whitener affects on the tooth by means of hydrogen peroxide itself or the oxygen radical that generated with Fe or Cu ion. Hence, these findings suggest that the penetration of peroxide from whitener applied to the tooth surface might cause some discomfort after whitening treatment.