Kinetic analysis for effect of initial substrate concentration on growth and secondary metabolite production in cultures of Nicotiana tabacum

Suspension cultures of tobacco cells were studied using airlift and rotary-drum bioreactors. The effect of initial concentrations of a major substrate, sucrose, on the growth and production of a secondary metabolite, phenolic compounds, was investigated. The dry weights and total concentrations of the phenolic compounds increased with the initial sucrose concentration in both bioreactors. Both bioreactors were found to have the same tendency for the effect of initial sucrose concentration. The structured model, presented previously was modified by considering that sucrose was hydrolyzed to glucose and fructose by an enzymatic reaction. The previous and the new models were applied to the above two sets of experimental data obtained with two bioreactors, independently. The hydrolysis of sucrose was elucidated to contribute slightly to the overall kinetics of growth and secondary metabolite production in these cultures. Furthermore, the levels of shear damage in each bioreactor were quantitatively compared based on the death rate constant, k_i, which is one of the model constants.     

Combining beta-core fragment and total oestriol measurements to test for Down syndrome pregnancies

Recent articles by Cuckle et al., Canick et al., and Isozaki et al. have evaluated urine beta-core fragment as a screening test for Down syndrome in second-trimester pregnancies. They found over four-fold elevation of beta-core fragment levels in Down syndrome pregnancies, and between 62 and 88 per cent detection of this trisomy at a 5 per cent false-positive rate. Urine beta-core fragment may be a superior screening test for Down syndrome pregnancies. In the present study, urinary total oestriol has been evaluated as a marker to use in combination with beta-core fragment in screening for Down syndrome pregnancies. The two markers were evaluated separately in relation to the urine creatinine concentration. To amplify screening performance, we evaluated the ratio of beta-core fragment to total oestriol levels (creatinine-independent). beta-core fragment and total oestriol levels were determined (normalized to creatinine, ng/mg creatinine) in urine samples from 480 unaffected and 12 Down syndrome pregnancies, collected consecutively at a single prenatal diagnosis centre. The median beta-core fragment level in Down syndrome cases was 4.5 MOM. Fifty-eight per cent of Down syndrome cases had beta-core fragment levels exceeding the 95th centile of unaffected pregnancies. The median total oestriol level in Down syndrome cases was 0.33 MOM. Forty-two per cent of Down syndrome cases had total oestriol levels exceeding the 95th centile of unaffected pregnancies. We investigated the ratio of the two determinants (beta-core fragment, ng/ml divided by total oestriol, ng/ml) in our sample set. The median beta-core fragment:total oestriol ratio in Down syndrome cases was 13 MOM. Seventy-five per cent of Down syndrome cases had a ratio exceeding the 95th and the 99.5th centile of unaffected pregnancies. Total oestriol complements beta-core fragment in urine screening for Down syndrome pregnancies. A test measuring the ratio of the two urine determinants may be a significant improvement over current serum methods for detecting Down syndrome.     

Analytical studies of four-inch pipe whip tests under BWR LOCA conditions

The purpose of pipe rupture studies in JAERI is to perform model tests on pipe whip, restraint behavior, jet impingement and jet thrust force, and to establish a computational method for analyzing these phenomena. This report presents the analytical results of 4-inch pipe whip tests under BWR LOCA conditions.Dynamic response analyses were performed using the general-purpose finite element program ADINA. The test pipe was modelled by straight beam elements and the four restraints were modelled by a single truss element. The analytical results were compared with the experimental results. Impact time and maximum total restraint force showed good agreement with experimental results. On the other hand, pipe strain and pipe deflection could not be predicted so well. The reason for this is that the sliding of the restraint during the pipe whip movement cannot be considered in the analyses.     

Expression of recombinant human thyrotropin receptor in myeloma cells

Starting with a previously isolated cDNA for human thyrotropin receptor (TSHR), we established a transformed myeloma cell line, SP56, which expresses human TSHR on its cell surface. Binding analysis showed that SP56 bears 1.1 x 10(5) TSHR per cell with a Kd of 2.2 x 10(-10) M. Using the purified cellular membrane, we established a TSH binding inhibition immunoglobulin (TBII) assay for autoantibodies against TSHR. We compared it with the TBII assay utilizing porcine thyroid membranes expressing porcine TSHR, which has been widely used for TBII assay, by using 96 serum samples from patients with autoimmune thyroid disease and normal individuals. Our TBII assay was more sensitive than the one using porcine TSHR: of 38 sera of patients which were judged negative for autoantibodies to TSHR (TBII value below 10%) by the latter assay, 28 were positive (above 20%) in our assay. By using a perfusion culture system, we obtained as many as 3 x 10(10) SP56 cells, from which 3,450 mg protein of the membrane could be purified; this is sufficient for 15,000 assays. The results indicate that the membrane of the myeloma cell line SP56 is more suitable for use in the TBII assay than the porcine thyroid membrane, in terms of sensitivity to autoantibodies against TSHR in human sera.     

Myopathology of the intrinsic laryngeal muscles in neurodegenerative diseases, with reference to the mechanism of vocal cord paralysis

To investigate the mechanism of the vocal cord abductor paralysis (VCAP) in the neurodegenerative diseases, the intrinsic laryngeal muscles (the crycothyroid, the interarytenoid, and the posterior crycoarytenoid muscles) from 41 autopsied cases were histologically examined: 10 cases of amyotrophic lateral sclerosis (ALS), 10 of Parkinson's disease (PD), 9 of multiple system atrophy (MSA), 4 of Machado-Joseph disease (MJD), 4 of progressive supranuclear palsy (PSP), 1 of familial amyloidotic polyneuropathy (FAP), and 3 of cerebrovascular diseases as a control. According to the distribution of the neurogenic changes among above-described three intrinsic laryngeal muscles, three forms were raised: 1. The totally paralytic form showing that all the three muscles developed neurogenic atrophy. This form includes ALS, MJD, and FAP. 2. The posterior muscle-paralytic form showing that only the posterior crycoarytenoid muscle was selectively involved. This form includes MSA. 3. The nonparalytic form showing no morphological abnormalities in any of the intrinsic laryngeal muscles. This type includes PD and PSP. In this nonparalytic form, supranuclear mechanism such as pyramidal or extrapyramidal tract involvement may cause VCAP through the increased laryngeal muscles tone. Considering that VCAP can be seen in any of the above-described forms, our results indicate that the mechanism of VCAP is different among the neurological disorders.     

Autoantibodies detectable in the sera of silicosis patients. The relationship between the anti-topoisomerase I antibody response and HLA-DQB1*0402 allele in Japanese silicosis patients

Eighty-one Japanese silicosis patients and 66 healthy volunteers were analyzed for autoantibodies by ELISA, and HLA-genotyping using the PCR-RFLP method was performed. Anti-topoisomerase I (anti-topo I) autoantibodies were detected in seven patients without any clinical features of autoimmune diseases such as progressive systemic sclerosis (PSS), although anti-topo I have been mostly reported in PSS patients. Antibodies directed to RNP, ssDNA, dsDNA and cent-B were not detected among the anti-topo I positive patients. The indirect immunofluorescent staining pattern of Hep-2 cells with the sera of anti-topo I positive silicosis patients demonstrated the typical mode of anti-topo I autoantibodies observed in the patients with PSS. The allelic frequency of HLA-DQB1*0402 was significantly higher in anti-topo I positive patients (28.6%) than in anti-topo I negative patients (1.5%, P < 0.001) or healthy controls (0.8%, P<0.001). HLA-DQB1*0301, DQB1*0601 and DPB1*1801 alleles were more frequently detected in anti-topo I positive patients than in the patients without anti-topo I or in healthy volunteers, but no significant difference was observed. DQB1 allele is associated with the induction of anti-topo I autoantibodies in Japanese silicosis patients, but the allele is not the same as in Caucasian PSS patients. Another allele (DQB1*0402) plays an important role in Japanese silicosis patients. The most important factor to induce anti-topo I autoantibodies seems not to be the type of alleles themselves, but the position of some specific amino acid residues in the DQ beta first domain. These findings will be useful for preventing occupational autoimmune diseases.     

Melting attack of solid plates by a high temperature liquid jet — effect of crust formation

When a molten UO₂ jet impinges on a steel structure in a reactor vessel during a severe accident, the erosion rate of the steel by the molten UO₂ jet is expected to be limited considerably by a UO₂ crust layer forming on a molten steel substrate at the jet/steel plate interface. A series of simulation experiments was performed to study the melting behavior of solid plates by high temperature liquid jets and the effects of crust forming at jet/structure interface. In the first series of experiments, salt (NaCl) was selected as the jet material and tin (Sn) as the solid structure. The experiments were conducted with varying the jet diameter (10 30 mm) and jet temperature (900 1100°C). The jets were accelerated to a range of 3 5 m/s at the nozzle outlet by gravitational force and impinged perpendicularly to the solid plate underneath. Furthermore, to check the effects of the thermo-physical properties on the erosion behaviors, preliminary experiments were performed by using a molten Al₂O₃ jet ( 2200°C) impinging on stainless steel plate at room temperature. The erosion rates obtained in the present experiments were far less than the values predicted by an analytical solution that neglects the existence of a crust layer and its thermal effects. With the inclusion of the crust behavior in the model, the experimental results were predicted fairly well. From the present experiments, a Nusselt number of the turbulent heat transfer, which takes into account simultaneous melting and freezing in the impingement region of a molten jet, is correlated by a Reynolds number and a Prandtl number as follows: Nu_m = 0.0033 Re---Pr.In conclusion, the existence of a crust layer plays an important role in the erosion process of a solid plate by the molten fuel jet with high melting point as in a reactor situation.     

Big insulin-like growth factor II-producing hepatocellular carcinoma associated with hypoglycemia

A 76-year-old female with a hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC) was hospitalized because of fasting hypoglycemia. Her sera contained a low concentration of immunoreactive insulin and insulin-like growth factor (IGF)-I, while the IGF-II level was normal. However, most of the IGF-II consisted of the high molecular weight form (big IGF-II). The tumor tissue contained fetal type of IGF-II mRNA (6.0 kb). Furthermore, we found that one of the four patients examined with HCV-related HCC had big IGF-II in serum. This indicates that non-islet cell tumor hypoglycemia (NICTH) in HCV-related HCC might be accompanied by production of big IGF-II by the tumor.     

Serum human hepatocyte growth factor (hHGF) is elevated in patients with metastatic gastric carcinoma

BACKGROUND/AIMS: Hepatocyte growth factor (HGF) is a stromally derived protein growth factor that modulates epithelial cell proliferation and motility. HGF may therefore be involved in tumor progression. METHODOLOGY: We measured the immunoreactive (ir)-HGF concentration in the sera of 56 patients with human gastric carcinoma, using an enzyme-linked immunosorbent assay and evaluated its association with clinical and histopathologic factors. Clinical stages were classified in accordance with The General Rules for TNM Classification (International Union Against Cancer). RESULTS: The serum hHGF concentrations from patients with gastric carcinoma significantly increased with increasing pathologic tumor grades. Also, there were significantly higher concentrations in patients with nodal metastasis compared to patients without metastasis. The serum hHGF concentrations in patients with liver metastasis were significantly higher than in patients without liver metastasis. CONCLUSIONS: These data show that the serum hHGF concentration is elevated in gastric carcinoma patients with liver and nodal metastasis. In multivariate analysis, the serum hHGF concentration was found to be the most important independent factor in predicting overall survival. Thus, the current results suggest that HGF may be important in the progression and metastasis of gastric carcinoma.     

Endoscopic evaluation of the depth of invasion of the depressed type early gastric cancer and apparently early advanced cancer in cases of a peptic ulcer within the cancer lesion

Depressed-type early and apparently early advanced cancers were divided into two groups according to whether or not an associated peptic ulcer was present within the cancer lesion, and the accuracy of preoperative endoscopic diagnosis was reviewed. As a result it was found that evaluation of depth of invasion was more difficult in the ulcer [UL(+)] group than in the group without an ulcer [UL(-)]. In the UL(+) group it was difficult to identify endoscopic findings which indicate depth of invasion at the tip of the folds, the volume of cancer invasion of the deeper layers was small, and there many cases of invasion of deeper layers in a portion of the margins of the peptic ulcers.