Reconstruction of a large DNA fragment from coinjected small fragments by homologous recombination in fertilized mouse eggs.
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Biomedical subjects
Publications and source records attributed to K Shimoda.
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Granulocyte colony-stimulating factor (G-CSF) in the cerebrospinal fluid from patients with meningitis was measured by our modified enzyme-linked immunosorbent assay for G-CSF. The minimal detection level was 20 pg/mL G-CSF. In patients with bacterial meningitis, the G-CSF levels in the cerebrospinal fluid were extremely elevated, showing a mean value of approximately 1,500 pg/mL. On the other hand, G-CSF levels in the cerebrospinal fluid from 67% patients with aseptic meningitis were moderately increased, showing a mean value of about 80 pg/mL, whereas G-CSF levels in 33% samples remained undetectable. The G-CSF levels and neutrophil counts in the cerebrospinal fluid were proven to be related by Spearman's rank correlation coefficient analysis (r = .724). These elevations of G-CSF levels at inflammation sites associated with bacterial meningitis may indicate that G-CSF plays an important role in the combat of bacterial infections.
Severe septic complications account for the high mortality of patients with esophageal cancer. We examined the levels of immunoglobulins and complements together with infection-related complications in a large number of patients. Enhancements of IgG, IgA, C3, C4, and CH50 were evident in patients with esophageal cancer and were more predominant compared to findings in cases of gastric cancer. Average levels of IgG and IgA immediately before surgery were significantly higher in esophageal cancer patients with postoperative septic complications than in those without such problems. Preoperative radiation therapy and total parenteral nutrition did not significantly alter the levels of immunoglobulins and complements. It would thus appear that the enhancement of IgG and IgA is associated with the occurrence of infectious complications following surgery for patients with esophageal cancer.
Correlations between defective cell-mediated immunity (CMI) and infections following surgery for esophageal cancer were evaluated. Peripheral lymphocytes, T cells, B cells, PHA transformation, and PPD skin test were measured in 81 patients with esophageal cancer, 58 with gastric cancer, and 50 healthy controls. The depression of CMI was predominant to a similar extent in patients with esophageal cancer and in those with gastric cancer. The average level of PHA transformation immediately before surgery was significantly lower in the esophageal cancer patients with fatal septic complications than in those without such problems. Although preoperative radiation therapy markedly depressed the levels of the four parameters, this association was also noted in 28 patients not given radiation. It thus appears that PHA transformation may be valuable in the prediction of fatal septic complications after major surgery in patients with esophageal cancer.
The expression of ras oncogene product p21 was examined in 45 paraffin-embedded sections of primary advanced colorectal cancers, using the anti-v-H-ras p21 monoclonal antibody Y13-259. Fourteen of these specimens (31 percent) were stained positively. The incidence of lymphatic vessel invasion of cancer cells and lymph node metastasis correlated statistically with the overexpression of ras p21. The depth of invasion and incidence of liver metastasis in the p21-positive group were more prominent than in the p21-negative group. Statistically significant differences were evident in operative curability and clinical stage at initial surgery and in the long-term survival rate between these groups (P less than 0.05). We propose that ras p21 overexpression may serve as a marker to predict the prognosis of colorectal cancer.
Lymphoscintigraphy (LS) and computed tomography (CT) were used preoperatively to assess nodal involvement in 23 patients with esophageal cancer. LS predicted nodal involvement with a high sensitivity and low specificity since false positives were frequent while in contrast, CT predicted it with high specificity and low sensitivity. The combination of LS and CT improved sensitivity by up to 87 per cent but not specificity. The positive image of nodes in LS was associated with malignant cell involvement and/or reactive changes in the nodes, especially sinus histiocytosis and germinal center hyperplasia. When cervical nodes were involved, bilateral images of lymph flows were often lost and by using the images of lymph flow as well as those of the nodes, the combination predicted cervical nodal involvement with a sensitivity of 83 per cent and a specificity of 86 per cent. Thus, the combination of LS and CT is beneficial for preoperatively estimating cervical nodal involvement in esophageal cancer.
Based on the findings that the enhancement of serum alpha 2-macroglobulin (A2M) is associated with the occurrence of infectious complications following surgery in patients with esophageal cancer, we examined possible factors which could contribute to the alterations of serum acute phase protein levels in patients with this disease. A multiple linear regression analysis was made for 71 patients with esophageal cancer and 58 with gastric cancer. In patients with esophageal cancer, protein calorie malnutrition (PCM) and age factors more strongly contributed to the alteration of 6 acute phase protein levels than did the malignant tumor when compared to those with gastric acner. PCM was negatively associated with A2M levels while it was positively associated with alpha 1-acidglycoprotein (A1AG) and haptoglobin (Hp) levels. Age did not contribute to the A2M levels but did have a negative effect on the Hp, ceruloplasmin (Cp) and fibronectin (Fn) levels. On the other hand, the malignant tumor was positively related only to the A1AG levels. Since none of these factors contributed to the elevation of A2M levels, it is suggested that the presence of chronic infection might be a factor contributing to the A2M increase which was associated with the occurrence of postoperative infectious complications in patients with this disease.
Severe septic complications are the major cause of operative mortality in patients with esophageal cancer. We examined the levels of acute phase proteins together with infection-related complications after surgery in a large number of patients with esophageal cancer and compared them with a group of patients with gastric cancer and healthy controls. Elevations in alpha 1-antitrypsin, alpha 1-acidglycoprotein, haptoglobin and ceruloplasmin were evident in patients with esophageal cancer, being more predominant when compared to the findings in patients with gastric cancer. Although the mean levels of alpha 2-macroglobulin were not significantly elevated in either patients with esophageal cancer or those with gastric cancer, the average level immediately prior to surgery was higher in esophageal cancer patients with postoperative septic complications than in those without any such problems. Preoperative radiation therapy and total parenteral nutrition did not significantly alter the levels of acute phase proteins. It would thus appear that the elevation in alpha 2-macroglobulin is associated with the occurrence of infectious complications following surgery in patients with esophageal cancer.
Ren-shen-yang-rong-tang (Japanese name: Ninjin-yoei-to, NYT) is a traditional Chinese herbal medicine. Leukocytosis and elevated levels of colony-stimulating factor (CSF) in peripheral blood were found previously after the administration of this compound to mice. In this study, human peripheral blood mononuclear cells (PBMC) were cultured in the presence of NYT in vitro, and the levels of granulocyte-macrophage CSF (GM-CSF) and granulocyte CSF (G-CSF) in the supernatant of cultured PBMC were measured using a sensitive enzyme-linked immunosorbent assays. NYT significantly (P less than 0.01) augmented GM-CSF production but not G-CSF production by PBMC in vitro.
A human lung squamous cell carcinoma cell line designated KSNY was established from a patient suffering from marked neutrophilia and polyclonal hyper-gamma-globulinemia. In our previous report, we demonstrated colony-stimulating activities in the culture supernatant of this cell line. To determine the exact molecules for the activities, we investigated the gene expression of various cytokines in KSNY cells and showed the mRNA expression of both granulocyte colony-stimulating factor (G-CSF) and interleukin-6 (IL-6). We also detected substantial amounts of G-CSF and IL-6 in the culture supernatant with sensitive enzyme-linked immunosorbent assays (ELISA). The amplification of the gene locus for G-CSF, but not for IL-6 was shown by Southern blot analysis. Furthermore, we also showed that the mRNAs for G-CSF and IL-6 were relatively stable in KSNY cells. These findings are thought to be related to the constitutive production of the cytokines in KSNY cells.
The level of granulocyte-macrophage colony-stimulating factor (GM-CSF) in the cerebrospinal fluid from 14 infants and children with meningitis and 6 patients who suffered other diseases besides meningitis was measured by our sensitive enzyme linked immunosorbent assay for GM-CSF. The minimal detection level of GM-CSF was 40 pg/ml. Six of 9 patients (67%) with aseptic meningitis had detectable GM-CSF in cerebrospinal fluid and the concentrations of GM-CSF ranged from 49 to 114 pg/ml (mean 72 pg/ml), whereas none of 5 patients with bacterial meningitis or 6 patients with other diseases besides meningitis had detectable GM-CSF levels. There was no clear correlation between the GM-CSF levels in cerebrospinal fluid and the leukocyte count in either peripheral blood or cerebrospinal fluid, or the concentration of protein or glucose in cerebrospinal fluid.
The stability of pathogenic bacteria from laboratory animals was investigated in various transport media at different temperatures. Bordetella bronchiseptica and Salmonella typhimurium survived for 8 days in phosphate-buffered saline (PBS, pH 7.0) at 37, 24, 4 and -20 degrees C; Brucella canis at 24, 4 and -20 degrees C; Corynebacterium kutscheri at 4 and -20 degrees C; and Pseudomonas aeruginosa at all but -20 degrees C. A marked decrease in numbers of Pasteurella multocida and Past. pneumotropica was observed in PBS at all temperatures. Skimmed milk in PBS improved the survival of Pasteurella spp. and Ps. aeruginosa at -20 degrees C. Neither glycerin, ascorbic acid nor sodium thioglycollate improved survival. The numbers of viable B. canis, Ps. aeruginosa and S. typhimurium were maintained in blood or faecal specimens held for 8 days at 4 degrees C. These results indicated that transport in PBS at 4 degrees C was the only method satisfactory for all species of pathogenic organisms tested, but Pasteurella spp. were the most difficult to maintain.
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In order to examine the ability of R plasmid-bearing Escherichia coli strains to colonize in the mouse alimentary tract, an R plasmid-positive (R(+)) E. coli strain and its R plasmid-negative (R(-)) counterpart were together inoculated into the streptomycin-treated mouse alimentary tract, and the numbers of fecal E. coli strains were enumerated. The numbers of R(+) strains were always at the level similar to or lower than those of their counterparts and rapidly decreased in the fecal population. However, when R plasmids, which were originated from a cryptic plasmid of the host E. coli strain, were utilized, an R(+) strain dominated over its R(-) counterpart during the experimental period. These experimental results indicated that the relationship between the host strain and R plasmids affected the ability of the host strain to colonize in the alimentary tract.
We performed lipiodolization and immunochemotherapy for recurrent pelvic tumor of sigmoid colon cancer using an infuserport which was implanted and connected to the catheter placed in the right internal iliac artery. Following lipiodolization, the level of CEA (980 ng/ml) decreased to within the normal range. MRI showed necrotic change and regression (more than 50%) of tumor. DSA revealed disappearance of tumor neovascularity. No serious side effect but skin erosion in the gluteal region was encountered after lipiodolization. This result suggests that lipiodolization is worth performing in further clinical trials for pelvic tumor.
Use of microangiography is now essential for the study of microcirculation in various organs. Renal microangiographic studies have been reported in rats, rabbits, dogs, human beings, and mice. However, we could not find any report on use of the technique in cattle, despite high incidence of renal disease in that species. The perfusion technique used in mice was improved over that of our previous report, and was applied to normal and diseased bovine kidneys. For the microangiographic technique, composition of the contrast medium, pressure of the injection, duration of perfusion, and washing of kidneys with heparinized saline solution before perfusion are important. In cattle, 1- to 2-mm-thick sections of the kidneys were generally necessary to observe renal vasculature: arcuate and interlobular arteries, afferent arterioles, and glomerular capillaries. In normal bovine kidneys, the angiographic and microangiographic findings were easily recognized as normal, compared with those of normal mice. In affected bovine kidneys, which histologically represented glomerulonephritis and pyelonephritis, angiography and microangiography revealed corresponding findings.
We have separated circulating HBeAg into small and large molecular forms by agarose gel electrophoresis and analyzed the relationship between the two forms and other clinical features of chronic hepatitis B, especially in regard to liver cell damage. The large HBeAg accounted for 7.3% +/- 3.4% of serum HBeAg in 9 subjects with normal liver histological findings or nonspecific reactive hepatitis, 38.0% +/- 27.8% in 32 patients with chronic persistent hepatitis and 65.6% +/- 23.3% in 21 patients with chronic active hepatitis (p less than 0.01). A positive correlation was seen between the height of aminotransferase elevations and the percentage of large HBeAg. Three patients who progressed from histologically normal liver or nonspecific reactive hepatitis to chronic active hepatitis had dramatic increases in the percentage of large HBeAg. The finding that the presence of large HBeAg in serum correlated with the severity of hepatitis suggests that HBeAg may play an important role in determining the degree of liver injury in chronic hepatitis B.
We experienced a long-term survived case with unresectable Stage 4 esophageal carcinoma well responded to chemoradiotherapy of combination of cisplatin (C), vindesine (V) and peplomycin (P) with radiotherapy (R). The patient was a male, aged 53, and diagnosed to have carcinoma, 8 cm in length, located at the IuCe region of the esophagus and right subclavicular lymph node metastasis on admission. He received radiotherapy at total dose of 60 Gy for the primary and metastatic lesions, and also 2 cycle CVP chemotherapy concomitantly. The mass of the primary lesion endoscopically changed to a flat area with small erosion while the right subclavicular metastatic node disappeared and was not found on CT after the end of the treatment. Objective response of this case was determined to the partial response since carcinoma cells were still found at the flat lesion. After discharge, 9 cycles of CVP therapy were further given in 3-year periods. At present, the patient is alive well although a part of the primary esophageal lesion is proved to have malignancy on biopsy.