Effects of gelonin immunoconjugate of monoclonal antibody MSN-1 to endometrial adenocarcinoma on antigen-producing tumor cells in vitro

Missile therapy, which destroys cancer cells specifically, has been considered to be an effective modality for treatment of carcinoma. We have developed a monoclonal antibody MSN-1 (immunoglobulin class: IgM), of which the immunogen is the endometrial adenocarcinoma cell line SNG-II, which strongly reacts with endometrial adenocarcinomas. We describe an immunoconjugate consisting of the MSN-1 and a plant hemitoxin named gelonin which has revealed to assume selective cytotoxicity against the SNG-II in a colony formation assay in vitro. The results of our study suggest that the 'inhibitory concentration' or IC50, of the MSN-1-gelonin immunoconjugate against the SNG-II was 188 fold that of gelonin alone. These results indicated that the MSN-1-gelonin immunoconjugate exhibited highly selective cytotoxicity to endometrial adenocarcinoma, which expressed an epitope against the MSN-1, and it is suggested that the MSN-1-gelonin immunoconjugate has possibility of clinical application to treatment of endometrial adenocarcinomas.     

Production and characterization of a monoclonal antibody (MSN-3) for uterine endometrial and endocervical adenocarcinoma

A monoclonal antibody (MSN-3) was raised using HEC-108 cells derived from poorly differentiated endometrial carcinoma as the immunogen. The immunoglobulin subclass of MSN-3 was IgGr1. The target antigen of MSN-3 was a protein with a molecular weight of 77 kDa, and it was shown to be localized in the cytoplasm. MSN-3 only reacted with 14% of normal proliferative endometrium cells, but it showed a high positivity rate of 66% for endometrial carcinoma. The target antigen of MSN-3 increased as endometrial cells became more malignant, and the possibility of changes in localization was also suggested. Moderately and poorly differentiated endometrial carcinoma showed a high positivity rate for MSN-3. MSN-3 reacted rarely or not at all with normal cervical glandular tissue, but the positivity rate for cervical adenocarcinoma (especially endocervical adenocarcinoma) was a high rate of 59%. The patterns of staining of endocervical adenocarcinoma by MSN-3 included diffuse staining of the whole cytoplasm and not only that near the glandular lumen, as well as staining of the basal cytoplasm. Changes in the localization of the target antigen were clearly associated with carcinogenesis of the cervical glandular cells. The MSN-3-positive rate was high in patients with lymph node metastasis and vascular invasion. Among the staining patterns, the basal and diffuse patterns tended to increase with malignacy. The basal pattern of staining was characteristic of MSN-3, suggesting that it might assist in the diagnosis of cervical adenocarcinoma.     

Antimicrobial susceptibility surveillance of obligate anaerobic bacteria in the Kinki area

Obligate anaerobes exist as resident flora in various sites in humans, but they are also emphasized as endogenous causative microorganism of infections. We performed surveillance to understand the trend of drug susceptibility in obligate anaerobic bacteria in the Kinki area of Japan. In the experiment, we used 156 obligate anaerobe isolates collected from 13 institutions that participated in the Study of Bacterial Resistance Kinki Region of Japan. MALDI Biotyper was used to identify the collected strains, and among the 156 test strains, those that could be identified with an accuracy of Score Value 2.0 or more included 6 genera, 30 species, and 144 strains (Bacteroides spp. 77 strains, Parabacteroides sp. 2 strains, Prevotella spp. 29 strains, Fusobacterium spp. 14 strains, Porphyromonas spp. 2 strains, and Clostridioides difficile 20 strains), and they were assigned as subject strains for drug susceptibility testing. The drug susceptibility test was carried out by broth microdilution method using Kyokuto Opt Panel MP ANA (Kyokuto Pharmaceutical Industrial Co., Ltd., Tokyo, Japan) and judged according to CLSI criteria. As a result, Bacteroides and Parabacteroides species showed good sensitivities to tazobactam-piperacillin, imipenem, metronidazole and chloramphenicol, and low sensitivities to ampicillin, cefoperazone and vancomycin. Prevotella species showed good sensitivities to sulbactam-ampicillin, tazobactam-piperacillin, cefmetazole, imipenem, doripenem and metronidazole. Susceptibility rates to other drugs were slightly different depending on the bacterial species. Both Fusobacterium spp. and Porphyromonas spp. showed high sensitivities to many drugs. C. difficile was highly sensitive to vancomycin and metronidazole, having MIC₉₀s of 0.5 μg/mL and ≤2 μg/mL, respectively.     

A prospective cohort study of ALI/ARDS in the Tohoku district of Japan (second report)

Purpose  

We previously reported a study of systemic inflammatory response syndrome (SIRS) cases in the Tohoku district of Japan in which the patients showed a 30-day mortality from acute lung injury/acute respiratory distress syndrome (ALI/ARDS) of about 20%. Cases in which chest X-ray findings did not meet ALI/ARDS criteria were diagnosed as acute hypoxemic respiratory failure (AHRF), but about 50% of these patients progressed to ALI/ARDS. The objective of this study was to verify the findings obtained in the earlier study and to gain further insights into the pathognomonic symptoms of AHRF associated with SIRS.     

Brain trauma induces expression of diacylglycerol kinase ζ in microglia

Diacylglycerol kinase (DGK) is an enzyme which phosphorylates a second messenger diacylglycerol and consists of a family of isozymes that differ in terms of structural motifs, enzymological property, and cell and tissue distribution. One of the isozymes, DGKζ was originally shown to be expressed in various kinds of neurons under physiological conditions. However, we unexpectedly found that under pathological conditions, such as cerebral infarction, DGKζ-immunoreactivity is detected in non-neuronal cells, although it remained to be elucidated in detail which cell types are responsible for the induced expression of DGKζ in this setting. To further elucidate functional implications of DGKζ in non-neuronal cells we performed detailed immunohistochemical analysis of DGKζ using rat brain cryoinjury model. As early as 1 h after cryoinjury, DGKζ-immunoreactivity was greatly decreased in the afflicted cerebral cortex and almost disappeared in the necrotic core. On day 7 after cryoinjury, however, DGKζ-immunoreactivity reappeared in this area. DGKζ-immunoreactivity was clearly detected in Iba1-immunoreactive cells of an oval or ameboid shape in the scar region, which represent activated microglia and/or macrophages. On the other hand, DGKζ-immunoreactivity was not detected in Iba1-immunoreactive, resting microglia of ramified and dendritic configuration in the intact cortex. Furthermore, DGKζ-immunoreactive cells were also positive for a microglia marker GLUT5 in the scar region, but never for an astrocyte marker GFAP. Taken together, the present study reveals that DGKζ is induced in activated microglia in brain trauma, suggesting the functional significance of DGKζ in this process.     

Changes of beta-1,4-galactosyltransferase with the development of endometrial cancer.

To elucidate the mechanism of the type 2 carbohydrate chain accompanying the development of endometrial cancer, we studied the expression of beta-1,4-galactosyltransferase (beta-1,4GT) in normal endometrial and endometrial cancer tissues. An immunohistochemical study revealed that beta-1,4GT was diffusely positive in the cytoplasm of endometrial cancer cells, and that the level of its expression was increased compared with that in normal endometrium. Also, beta-1,4GT mRNA corresponding to 4.7 kb was very low in normal endometrium, while an intense signal was detected in endometrial cancer. Our results suggest that the increase of beta-1,4GT contributes to the expression of the type 2 carbohydrate chain in endometrial cancer.     

Purification and partial characterization of acetyl-coA synthetase in rat liver mitochondria

Acetyl-CoA synthetase (AceCS), which catalyzes the activation of acetate to produce acetyl-CoA, was found to have a much greater Km value for acetate in liver mitochondria than that in the heart mitochondria of rats, indicating that two different types of AceCS are located in the liver and heart mitochodria. Recently, Fujino et al. reported that mouse heart mitochondrial AceCS, designated AceCS2, was expressed in a wide range of tissues, however, it was apparently absent from the liver. In this study, liver mitochondrial AceCS activity, but not heart AceCS2, was greatly induced in di(2-ethylhexyl)phthalate (DEHP)-treated rats. We purified and characterized the rat liver mitochondrial AceCS. The molecular mass of the enzyme estimated by SDS-PAGE was -58 kDa, which was quite different from that of the heart mitochondrial enzyme, AceCS2. The calculated Km value for the acetate of the partially purified liver enzyme was much greater, being about 100 times that of heart enzyme, AceCS2.     

Expression of glycolipids bearing Lewis phenotypes in tissues and cultured cells of human gynecological cancers.

Transformation-associated expression of Le(b) (Lewis antigen-b) or Le(Y) in human colorectal carcinomas has been well described. To examine the expression of glycosphingolipids (GSLs) bearing Lewis-phenotypes in human gynecological carcinoma-derived cells, we determined the concentrations of all GSLs. Although neither Le(b) nor Le(Y) was present in HEC-108 cells established from the poorly differentiated type of endometrial adenocarcinoma, other cell lines from moderately or well-differentiated types expressed either Le(b) or Le(Y), or both, at concentrations of 0.01 to 0.03 microg per mg of dry cells, which comprised 0.3 to 1.3% of the total GSLs. In the cervical and ovarian carcinoma-derived cell lines, Lewis phenotypes tended to be carried by nLc(4)Cer, which was accumulated in the cells without sialylation or fucosylation. These results indicated that expression of Le(b)- or Le(Y)-phenotypes was strongly dependent on the metabolic ability to supply the precursor GSLs. Both Le(b) and Le(Y) were successfully detected by monoclonal antibody MSN-1, which was a useful probe for the simultaneous detection of Le(b) and Le(Y). On application of MSN-1, either Le(b) or Le(Y) was detected in tissues from patients with well- and moderately differentiated types of endometrial adenocarcinoma at concentrations of 0.01 to 0.04 microg per mg of dry tissues, but not in the tissues of poorly differentiated type. Normal endometria at the follicular and luteal phases also contained the antigens, but the concentrations and the frequency of antigen expression were lower than those in the well- and moderately differentiated types of endometrial adenocarcinoma.     

Expression of Glycolipids Bearing Lewis Phenotypes in Tissues and Cultured Cells of Human Gynecological Cancers

Transformation-associated expression of Le^b (Lewis antigen-b) or Le^Y in human colorectal carcinomas has been well described. To examine the expression of glycosphingolipids (GSLs) bearing Lewis-phenotypes in human gynecological carcinoma-derived cells, we determined the concentrations of all GSLs. Although neither Le^b nor Le^Y was present in HEC-108 cells established from the poorly differentiated type of endometrial adenocarcinoma, other cell lines from moderately or well-differentiated types expressed either Le^b or Le^Y, or both, at concentrations of 0.01 to 0.03 μg per mg of dry cells, which comprised 0.3 to 1.3% of the total GSLs. In the cervical and ovarian carcinoma-derived cell lines, Lewis phenotypes tended to be carried by nLc₄Cer, which was accumulated in the cells without sialylation or fucosylation. These results indicated that expression of Le^b- or Le^Y-phenotypes was strongly dependent on the metabolic ability to supply the precursor GSLs. Both Le^b and Le^Y were successfully detected by monoclonal antibody MSN-1, which was a useful probe for the simultaneous detection of Le^b and Le^Y. On application of MSN-1, either Le^b or Le^Y was detected in tissues from patients with well- and moderately differentiated types of endometrial adenocarcinoma at concentrations of 0.01 to 0.04 μg per mg of dry tissues, but not in the tissues of poorly differentiated type. Normal endometria at the follicular and luteal phases also contained the antigens, but the concentrations and the frequency of antigen expression were lower than those in the well- and moderately differentiated types of endometrial adenocarcinoma.